CA2241633A1 - Vitronectin receptor antagonists - Google Patents
Vitronectin receptor antagonists Download PDFInfo
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- CA2241633A1 CA2241633A1 CA002241633A CA2241633A CA2241633A1 CA 2241633 A1 CA2241633 A1 CA 2241633A1 CA 002241633 A CA002241633 A CA 002241633A CA 2241633 A CA2241633 A CA 2241633A CA 2241633 A1 CA2241633 A1 CA 2241633A1
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Abstract
Compounds of formulae (I-V) are disclosed which are vitronectin receptor antagonists and are useful in the treatment of osteoporosis.
Description
W O 97/24119 PCTrUS96/20748 TITLE
Vitronectin Receptor Antagonists FIELD OF THE INVENTION
This invention relates to pharm~eu~ically active compounds which inhibit the vitronectin receptor and are useful for the treatment o~ infl~mm~tion, cancer and cardiovascular disorders, such as atherosclerosis and restenosis, and ~ e:~e.s wherein bone resorption is a factor, such as osteoporosis.
BACKGROUND OF TH~ INVEI~TION
Integrins are a ~upelr~lllily of cell adhesion receptors, which are transmembrane glycoproteins expressed on a variety of cells. These cell surface adhesion receptors include gpIIb /IIIa, the fibrinogen receptor, and o~vM3, the 15 vitronectin receptor. The fibrinogen receptor gpIIb /IIIa is expressed on the platelet surface and it m~ tes platelet aggregation and the formation of a hemostatic clot at the site of a bleeding wound. Philips, et al., Blood., 1988, 71, 831. The vitronectin receptor av1~3 is expressed on a number of cells, including endothelial, smooth muscle, osteoclast, and tumor cells, and, thus, it has a variety of functions. The aV133 20 receptor expressed on the membrane of osteoclast cells mediates the bone resportion process and contributes to the development of osteoporosis. Ross, et al., J. Biol.
Chem., 1987, 262, 7703. The ocv~3 receptor expressed on human aortic smooth muscle cells ~tim~ tPs their migration into neointima, which leads to the formation of atherosclerosis and restenosis after angioplasty. Brown, et al., Cardiovascular 25 Res., 1994, 28, 1815. Additionally, a recent study has shown that a CCv~3 antagonist is able to proIrlote turnor regression by in.lu.~ing aEioptosi~. of angiogenic l~lood vessels. Brooks, et al., Cell, 1994, 79, 1157. Thus, agents that would block thevitronectin receptor would be useful in treating diseases m~ t~d by this receptor, such as osteoporosis, atherosclerosis, restenosis and cancer.
~ -- 1 --W O97/24119 PCT~US96/20748 The vitronectin receptor is known to bind to bone matrix proteins, such as osteopontin, bone sialoprotein and thrombospondin, which contain the tri-peptideArg-Gly-Asp (or RGD) motif. Thus, Horton, et al., Exp. Cell Res. lg91, 195, 368,disclose that RGD-cont:lining peptides and an anti-vitronectin receptor antibody(23C6~ inhibit dentine resorption and cell spreading by osteoclasts. In addition, Sato, et al., J. Cell Biol. 1990, 111, 1713 disclose that echicf~fin, a snake venom peptide which contains the RGD sequence, is a potent inhibitor of bone resorption in tissue culture, and inhibits attachment of osteoclasts to bone. Fisher, et al., Endocrinology 1993, 132, 1411, has further shown that echistatin inhibits bone resorption in vivo in the rat. BertoIini etal., J. Bone Min. ~es., 6, Sup. 1, S146, 252 have shown that cylco-S,S-N~C-acetyl-cy~leillyl-N~c methyl-argininyl-glycyl-aspartyl-penicillzlmin~ inhibits osteoclast ~tt:~hmt-nt to bone. EP 528 587 and 528 586 report substituted phenyl derivatives which inhibit osteoclast m~ tçA bone resorption.
Alig et al., EP 0 381 033, Hartman, et al., EP 0 540,334, Blackburn, et al., WO 93/08174, Bondinell, et al., WO 93/00095, Blackburn, et al. WO 95/04057, Egbertson, et al, EP 0 478 328, Sugihara, et al. EP 529,858, Porter, et al., EP 0 542 363, and Fisher, et al., EP 0 635 492 ~1icclc)se certain compounds that are useful for inhibiting the fibrinogen receptor. It has now been discovered that certain a~ iately substituted compounds are potent inhibitors of the vitronectin receptor.
In particular, it has been discovered that such compounds are more potent inhibitors of the vitronectin receptor than the fibrinogen receptor and such compounds contain a fibrinogen receptor antagonist template.
SUM[MARY OF THE INVENTION
This invention comprises compounds of the formula (I)-(V) and (XXI)-(XXII) as described hereinafter, which have pharmacological activity for the inhibition of the vitronection receptor and are useful in the treatment of infl~mm:-fion, cancer and cardiovascular disorders, such as atherosclerosis and restenosis, and ~lice~cçs wherein bone resorption is a factor, such as osteoporosis.
W O g7/24119 PCTrUS96120748 This invention is also a pharm~reutical composition comprising a compound according to formula (I)-(V) (XXI)-(XXII)and a ph~rm:~reutically carrier.
This invention is also a method of treating ~ e~es which are m~ t~d by " the vitronectin receptor. In a particular aspect, the compounds of this invention are 5 useful for treating atherosclerosis, restenosis, infl~mm~tion, cancer and diseases wherein bone resorption is a factor, such as osteoporosis.
DETAILI~D DESCRIPTION
This invention compri~es novel compounds which are more potent inhibitors 10 of the vitronectin receptor than the fibrinogen receptor. The co~ oullds of the instant invention comprise a fibrinogen receptor antagonist template that is linked to a nitrogen-cont~ining five-membered ring, which is optionally fused to an aromatic six-membered ring. The fibrinogen receptor antagonist template is substituted by an aliphatic substituent which contains an acidic moiety. It is preferred that about 15 fourteen intervening covalent bonds via the shortest intramolecular path will exist between the acidic group of the fibrinogen receptor antagonist template and the nitrogen of the optionally fused five-membered ring.
As used herein, the term "fibrinogen recel~lor antagonist template" means the core structure of a fibrinogen receptor antagonist, said core being substituted by an 20 acidic group and said core being linked to an organic group substituted with a basic nitrogen moiety. A fibrinogen receptor antagonist is an agent that inhibits the binding of fibrinogen to the platelet-bound fibrinogen receptor GPIIb-IIIa. It is an object of this invention that a fibrinogen receptor antagonist is converted to avitronectin receptor antagonist by replacing the organic group substituted with a 25 basic nitrogen moiety in a fibrinogen receptor antagonist with an optionally fused nitrogen-cont~ining five-membered ring, preferably an imidazole ring and, most preferably, a ben~imi~ ole ring.
W O 97/24119 PCTrUS96/20748 This invention conl~lises compounds of formula (I)~
W A Rb G
,~ /~ R' ~W--A
(I) or (Il) or (1ll) or R" Rc~N--W A
(IV) or (V) wherein:
W is - (CHRg)b-V'- or Ar;
A is a fibrinogen receptor antagonist template;
V' is CONR21 or NR2~CO;
GisNRe,SorO;
Rg is H, Cl 6alkyl, Het-CO 6alkyl, C3 7cycloalkyl-CO 6alkyl or Ar- CO 6alkyl;
R21 is Het-CO 6alkyl-U'-CI 6alkyl-~ C3 7cycloalkyl-CO 6alkyl-U'-CI 6alkyl-, or Ar-CO 6alkyl-U'-C I 6alkyl-;
U' is CONRf or NRfCO;
Rf is H, Cl 6alkyl or Ar-CI 6alkyl;
Re is H, Cl 6alkyl, Ar-CI 6alkyl, Het-CI 6alkyl, C3 7cycloalkyl-CI 6alkyl, (CH2)qOH or (CH2)kCO2Rg;
kisO, 1 or2;
q is 1 or 2;
bisO, 1 or2;~0 Rb and RC are independently sele.cte~l from H, Cl 6alkyl, Ar-CO 6alkyl, Het-CO 6alkyl, or C3 6cycloalkyl-CO 6alkyl, halogen, C1~3, ORf, S(O)kRf, CORf, NO2, N(Rf)2, Co(NRf)2, CH2N(Rf)2, or Rb and RC are joined together to form a five or six membered aromatic or non-aromatic carbocyclic or heterocyclic ring, optionally substituted by up to three substituents chosen W O 97/24119 PCT~US96/20748 from halogen, CF3, Cl 4alkyl, ORf~ S(O)kRf, CORf, CO2Rf OH, N02, N(Rf)2, CO(NRf)2, and CH2N(Rf)2, or methylenedioxy;
or a pharm~-~elltically acceptable salt thereof.
This invention also comprises compounds of formula (XXI)-(XXII):
I~I~ G
~ B A ~ /~ B A
~XXI) or (XXII) wherein:
B is a linking moiety of the form -(CHRg)a-U- (CHRg)b-V-;
A is a fibrinogen receptor antagonist template;
G is NRe, S or O;
Rg is H, Cl 6alkyl, Het-CO 6alkyl, C3 7cycloalkyl-C0 6alkyl or Ar- C0 6alkyl;
Rk is Rg, -C(O)Rg, or -C(O)ORf;
Ri is is H, Cl 6alkyl, Het-CO 6alkyl, C3 7cycloalkyl-C0 6alkyl, Ar- C0 6alkyl, Het-CO 6aL~cyl-U-C1 6alkyl-, C3 7cycloalkyl-CO 6alkyl-U'-Cl 6alkyl-, or Ar-CO 6alkyl-U'-CI 6alkyl- or Cl 6alkyl;
lS Rf is H, Cl 6alkyl or Ar-CI 6alkyl;
Re is H, Cl 6alkyl, Ar-CI 6alkyl, Het-CI 6alkyl, C3 7cycloalkyl-CI 6alkyl, (CH2)qOH or (CH2)kCO2Rg;
U, U' and V independently are absent or CO, CRg2, C(=CRg2), S(O)k, O, NRg, CRgORg, CRg(ORk)CRg2, CRg2CRg(ORk), C(O)CRg2, CRg2C(O), CON Ri N Ri CO OC(O), C(O)O, C(S)O, OC(S), C(S)NRg, NRgC(S), S(O)2NRg, NRgS(O)2, N=N, NRgNRg, NRgCRg2, NRgCRg2, CRg2O, OCRg2, C_C, CRg=CRg~ Ar or Het;
kisO, 1 or2;
~ qis I or2;
W O 97/24119 PCT~US96/20748 aisO, 1 or2;
bisO, 1 or2;
Rb and RC are independently selected from H, Cl 6alkyl, Ar-CO 6alkyl, Het-CO 6alkyl, or C3 6cycloalkyl-CO 6alkyl, halogen, CF3, oRf, S(o)kRf, CORf, N02, N(Rf)2, Co(NRf)2, CH2N(Rf)2, or Rb and RC are joined together to forrn a five or six membered aromatic or non-aromatic carbocyclic or ~ heterocyclic ring, optionally- substituted by up to three substituents chosen from halogen, CF3, Cl 4alkyl, ORf, S(O)kR~, CORf, C02Rf OH, N02, N(Rf)2, CO(NRf)2, and CH2N(Rf)2; or methylenedioxy;
10 or pharm~reuticSllly acceptable salts thereof.
Preferably, U' is CONRf or NRfCO.
Also included in this invention are pharm:3relltically acceptable addition salts, complexes or prodrugs of the compounds of this invention. Prodrugs are considered to be any covalently bonded carriers which release the active parent drug lS according to formula (I) in vivo. In cases wherein the compounds of this invention may have one or more chiral centers, unless specified, this invention includes each unique nonracernic compound which may be synth~o~i7ç~1 and resolved by conventional techniques. In cases in which compounds have unsaturated carbon-carbon double bonds, both the cis (Z) and trans (E) isomers are within the scope of 20 this invention. In cases wheleill compounds may exist in tautomeric forms, such as O OR' keto-enol tautomers, such as ~~ and b ~, and each tautomeric form is contemplated as being included within this invention whether existing in equilibrium or locked in one form by a~ iate substitution with R'.
The compounds of formula (I) - (V) and (XXI) - (XXII) inhibit the binding of vitronectin and other RGD-co..~ lg peptides to the vitronectin (ocv1~3) receptor.
Inhibition of the vitronectin receptor on osteoclasts inhibits osteoclastic boneresorption and is useful in the tre~tm~nt of ~ e~sç~ wherein bone resorption is associated with pathology, such as osteoporosis. Additionally, since the compounds W O 97/24119 PCTrUS96120748 of the instant invention inhibit vitronectin receptors on a number of different types of cells, said compounds would be useful in the tre~tm~n~ of infl~mm~tion and cardiovascular diseases, such as atherosclerosis and restenosis, and would be useful as anti-m~t~cta~ir and ~ntitllmt~r agents.
In a particuar embodiment, the compounds of this invention are of the formula (II), wherein Rb and Rc are joined to form an aromatic ring ct-nt~ining up to two nitrogen atoms. In a preferred embodiment Rb and Rc are joined to form an optionally substituted phenyl ring according to formula (IIa):
RY ~C /~ W-A
R (IIa) wherein G is N-R, S, CH or 0.
L N-CO
Suitably W is -(CHRg)aNRiCO- or ~~ , or, when G is CH, W is -CH2CH2NRiCO-wherein Ri is a methylene group :lt~hl-cl to G.
Preferably W is -CHRgNRiC0-.
Suitably Ri is H, Cl 6alkyl, C3 7cycloalkyl, Ar or Cl 6alkyl substituted by one to three groups chosen from halogen, CN, NRg2, ORg, SRg, C02Rg, and CON(Rg)2, Ar, Het or C3 7cycloalkyl. In particular, Ri is H, methyl, butyl, cyanomethyl, carboxymethyl, phenylethyl or ben7imi~ 701ylmethyl.
Suitably RX, RY and RZ are independently chosen from Cl 6alkyl, methoxy, 20 nitro, trifluoromethyl, fluoro, chloro, amino or Rx and RY are adjacent to one another and are joined to form a methylenedioxy group.
Preferably G is NRe.
Suitably Re is H, Cl 4alkyl, Ar, Het or Cl 4alkyl substituted by Ar or Het.
More suitably, Re is H, methyl or ben7.imi~1~7olylmethyl.
W O 97/24119 PCT~US96/20748 In another speeific embodiment, Rb and Rc form a six membered aromatic ring eon~ining one or two nitrogen atoms according to formulas (IIb-d):
N R~ \~W A ~ \~W A
(lIb) (IIc) (IId) 5 wherein G, RX and RY are as above for formula (IIa).
Specifically, the compounds of this invention are comprised of a nitrogen-cont~ining optionally fused five-m~mhered ring, a linking group W, and a fibrinogen reeeptor antagonist template A. In partieular, the fibrinogen reeeptor antagonist template A is as defmed in Bondinell, et al., WO 93/00095, publich~ocl January 7, 1993, of the sub-formula (VI):
D A'--A (VI) Al to As form an ~reeccihle ~ub~LiLuLed seven-membered ring, which may be saturated or unsaturated, optionally eontaining up to two heteroatoms ehosen from 15 the group of O, S and N wlle~ S and N may be optionally oxi~
Dl to D4 form an aeeessible substituted six membered ring, optionally cont~ining up to two nitrogen atoms;
R is at least one substituent ehosen from the group of R7, or Q-Cl 4alkyl, Q-C2 4alkenyl, Q-C24alkynyl, optionally substituted by one or more of =O, Rl 1 or 20 R7;
R* is H, Q-Cl 6alkyl, Q-C1 6oxoalkyl, Q-C2 6alkenyl, Q-C3 40xoalkenyl, Q-C3 40xoalkynyl, Q-C24alkynyl, C3 6eyeloalkyl, Ar or Het, optionally substituted by one or more of Rl l;
Q is H, C3 6eyeloalkyl, Het or Ar;
CA 0224l633 l998-06-26 PCTrUS96/20748 R7 is -COR8, -COCR'2R9, -C(S)R8, -S(O)mOR', -S(O)mNR'R", -PO(OR'), -PO(OR')2, -B(OR')2, -NO2 and Tet;
R8 is -OR', -NR'R", -NR'SO2R', -NR'OR', -OCR'2C(O)OR', -OCR'2OC(O)-R', -OCR'2C(O)NR'2, CF3 or AAl;
R9 is -OR', -CN, -S(O)rR', S(O)mNR'2, -C(O)R' C(O)NR'2 or -CO2R';
Rl I is H, halo, -OR12, -CN, -NR'RI2, -NO2, -CF3, CF3S(O)r, -CO2R', -CONR 2, Q-C0 6alkyl-, Q-Cl 6oxoalkyl-, Q-C2 6alkenyl-, Q-C2 6alkynyl-, Q-Co 6alkyloxy-, Q-C0 6alkylamino- or Q-C0 6alkyl-S(O)~;
R12 is R', -C(O)R', -C(O)NR'2, -C(O)OR15, -S(O)mR' or S(O)mNR'2;
Rl3 is R', -CF3, -SR', or -OR';
Rl4 is R', C(O)R', CN, NO2, SO2R' or C(O)oRl5;
Rls is H, Cl 6alkyl or Ar-Co 4alkyl;
R' is H, Cl 6alkyl, C3 7cycloalkyl-Co 4alkyl or Ar-Co 4alkyl;
R" is R', -C(O)R' or-C(O)OR15;
R"' is R" or AA2;
AAl is an amino acid attached through its amino group and having its carboxyl group optionally protected, and AA2 is an amino acid attached through its carboxyl group, and having its amino group optionally protected;
mis 1 or2;
nisOto3;
pisOor l;and tisOto2;or ph~ celltic~lly acceptable salts thereof.
With reference to formula (VI), suitably, Al is CRlRl, CRl, NRl, N, O or S(O)x;
A2 iS CR2R2~ CR2, NR2;
A3 is CR3R3, CR3, NR3, N, O or S(O)x;
g CA 02241633 l99X-06-26 W O 97/24119 PCTrUS96120748 A4 is CR4R4, CR4, NR4, or N;
A5 is CRsRs, CRs, NR5, N, O or S(O)x;
Dl-D4 are CRIl~ CR6 orN;
Rl and Rl are R* or R, or together are =O;
R2 and R2 are R*, R or =O;
R3andR3 areR*,Ror=O;
R4 and R4 are R*, R or =O; .
Rs and R5 are R*, R or =O; and xisOto2.
More suitably, Al is CRlRl, CR1, NRl, N, O or S; A2 is CR2R2, NR2 or CR2; A3 is CR3R3; A4 is CR4R4, CR4, NR4, or N; AS is CR5Rs, CR5, NR5, N, O;
Dl - D4 are CH; R2 or R4 are R; R3,R3 and R5,R5 are =0 or R*,H.
Preferably, Al is CHRl, CRl, NR", N or S; A2 is CR2 or CR2R2; A3 is CR3R3; A4 is CR4R4 or NR4; As is CR5Rs, and Dl - D4 are CH.
In one embodiment, Al is CRl, A2 is CR2, A3 is C=O, A4 is NR4 and As are CHRs.
In another embodiment, Al is NRl, A2 is CHCR2, A3 is CR3R3, A4 is NR4, and A5 are C=O.
In yet another embodiment, Al and A4 are C=O, A2 is NR2, A3 is CHR3 and 20 AS is NRs.
In a preferred embodiment, Al is NRI, A2 is CHR2, A3 is C--O, A4 is NR' and A5 is CHRs.
Representative sub-form~ of (VI) are given by each of formulas (VIa)-(VIi) below:
,R-I ~5N,FI,~R3, R R2 N R2 R?~R ' (VIa) (VIb) (VIc) W O 97/24119 PCT~US96120748 , R~
(VId) (VIe) (VIf) {~ ~ )~ R3 Rl R1 2 , R2 or R1 R1' (VIg) (VIh) (VIi) Specific embodiments of this invention wherein the fibrinogen receptor antagonist template A is of the sub-formula (VI) are named in the Examples.
Preferred compounds of this invention are:
5-[[[(Benzimidazol-2-yl)methyl]methylamino]carbonyl]-lH-ben7.imicl:~701e-2-aminoacetic acid;
(+)-2,3,4,5-Tetrahydro-7-[[[(ben7imi~i~7.ol-2-yl)methyl}-methylamino~carbonyl]-4-(3 ,3-dimethylbutyl)-3-oxo- 1 H- 1 ,4-benzodiazepine-2-acetic acid;
(S)-2,3,4,5-Tetrahydro-4-methyl-3-oxo-7-[[[(5-trifluoromethylb~n7.imi(1~7.ol-2-yl)methyl]methylamino]carbonyl]- 1 H- 1 ,4-benzodiazepine-2-acetic acid;
(S)-2,3,4,5-Tetrahydro-7-[[[(4,7-dimethoxyben7.imi~1~7.ol-2-yl)methyl]methylamino~carbonyl]-4-methyl-3-oxo- 1 H- 1 ,4-benzodiazepine-2-acetic acid;
(+)-2,3,4,5-Tetrahydro-7-[[[(benzimidazol-2-yl)methyl]amino]carbonyl]-4-(3 ,3-dimethylbutyl)-3-oxo- 1 H- 1 ,4-benzodiazepine-2-acetic acid;
~. (S)-2,3,4,5-Tetrahydro-7-[[[(4-methylb~n7.imicl~7.ol-2-yl)methyl~methylamino]carbonyl]-4-methyl-3-oxo- 1 H- 1 ,4-benzodiazepine-2-acetic acid;
S3-2,3,4,5-Tetrahydro-7-[[N-[(ben7.imill~7nl-2-yl)methyl]-N-(4-anninobutyl3amino]carbonyl]-4-methyl-3-oxo-lH-1,4-benzodiazepi.le-2-acetic acid;(S)-2,3,4,5-Tetrahydro-7-[[N-(ben ~.i ., ~idzl701-2-yl)methyl-N-(2-cyanomethyl)amino]carbonyl]-4-methyl-3-oxo-lH-1,4-b~.n70~ Ppine-2-acetic 5 acid;
(S)-2,3,4,5-Tetrahydro-7-[[[(be~7.imi~1~7.ol-2-yl)methyl]amino~carbonyl]-3-oxo-4-(4-phth~limiclobutyl)-lH-l~4-benzodiazepine-2-acetic acid;
4-[[r3-(Ren7.imitl~701-2-yl)propyl]amino]carbonyl]piperidine-1-acetic acid;
4-[[~3-(Ben7.imi~1~701-2-yl)propyl]amino]carbonyl]phenylacetic acid;
(S)-2,3,4,5-Tetrahydro-7-[[[(4-aza-5,7-dimethylben7.imi~ 7-1-2-yl)methyl3methylamino~carbonyl]-4-methyl-3 -oxo- 1 H- 1 ,4-benzodiazepine-2-acetic acid;
(~t)-2,3,4,5-Tetrahydro-7-[[[(bel~7.i,..i~1~7.ol-2-yl)methyl3methylamino~-carbonyl}-4-[2-(3 ,4-methylenedioxyphenyl)ethyl]-3-oxo- 1 H- 1 ,4-benzodiazepine-2-acetic acid;
(~)-2,3 ,4,5-Tetrahydro-7-[[[(ben7.imidz-701-2-yl)methyllamino]carbonyl]-4-(2-methoxyethyl)-3 -oxo- 1 H- 1 ,4-benzodiazepine-2-acetic acid;
(S)-2,3,4,5-Tetrahydro-7-[[[(bt~n7.imi~1~7.ol-2-yl)methyl]methylamino]-carbonyl]-4-methyl-3-oxo- 1H- 1 ,4-benzodiazepine-2-~f et~mi~le;
(+)-2,3,4,5-Tetrahydro-7-[[[[1-[(ben7.imi~1~7.ol-2-yl)methyl]ben7.imi(1~7.ol-2-yl]methyl]amino]carbonyl]4-methyl-3-oxo-lH-1,4-ben7.orliz~7epine-2-acetic acid;
(S)-2,3,4,5-Tetrahydro-7-[[[(be.n7.imi-1~7.~-1-2-yl)methyl]methylamino]carbonyl~-3 -oxo- 1 H- 1 ,4-benzodiazepine-2-acetic acid;
(S)-2,3,4,5-Tetrahydro-7-[[bis[(benzimidazol-2-yl)methyl]amino]carbonyl]-4-methyl-3-oxo- lH-1 ,4-benzodiazepine-2-acetic acid;
(+)-2,3,4,5-Tetrahydro-7-r[[(4-azaben7.imit1~701-2-yl)methyl~methylamino]carbonyl]-4-(3 ,3 -dimethylbutyl)-3-oxo- 1 H- 1,~
benzodiazepine-2-acetic acid;
(i~)-2,3,4,5-Tetrahydro-7-[[[(ben7.imi<1~7.Ql-2-yl)methyl]methylamino]carbonyl]-3-oxo-4-(2,2,2-trifluoroethyl)-lH-1,4-benzodiazepine-2-acetic acid;
CA 0224l633 l998-06-26 W O 97/24119 PCT~US96/20748 (~)-2,3,4,5-Tetrahydro-7-t[2-(ben7.imid~7.ol-2-yl)acetyl~amino]-S-oxo-4-(2-phenylethyl)- 1 H- 1 ,4-benzodiazepine-2-acetic acid;
, (+)-2,3,4,5-Tetrahydro-7-[[[(ben7.imi-1~7.ol-2-yl)methyl]amino]carbonyl]-3-oxo-4-(2,2,2-trifluoroethyl)- lH- 1 ,4-benzodiazepine-2-acetic acid;
" S (S)-2,3,4,5-Tetrahydro-7-~[[(5,6-difluoroberl7.imifl:~7.ol-2-yl)methyl]methylamino]carbonyl]-4-methyl-3-oxo- lH- 1 ,4-benzodiazepine-2-aceticacid;
(+)-2,3,4,5-Tetrahydro-7-[[bis[(ben7.imicl~7.ol-2-yl)methyl]amino]carbonyl]-3-oxo-4-(2-phenylethyl)- lH- 1 ,4-ben70~ 7.epine-2-acetic acid;
(S)-2,3,4,5-Tetrahydro-7-[[[~4-aza-5-methylben7imid:~7.ol-2-yl)methyl]arnino]carbonyl]-4-methyl-3-oxo- 1 H- 1 ,4-benzodiazepine-2-acetic acid;
(S)-2,3,4,5-Tetrahydro-4-methyl-7-[[[(4-nitroben7.imi(1~7.ol-2-yl)methyl]methylaminolcarbonyl]-3-oxo- lH- 1 ,4-benzodiazepine-2-acetic acid;
(+)-2,3,4,5-Tetrahydro-7-[[[(4-aza-S-methylben7.imi~ 7.ol-2-15 yl)methyl]amino]carbQnyl]-3-QxQ-4-(2,2,2-trifluoroethyl)- lH- 1 ,4-benzodiazepine-2-acetic acid;
(+)-4-~4-[[L( iH-Ben7.imidi~7.oi-2-yi)melhyljmethylamino]ccubonyl]phenyl]=3=
phenylbutanoic acid;
(+)-3-[[[4-(4-Azabe.,~ 7.Ql-2-yl)butanoyl]glycyl]amino]-4-pentynoic 20 acid;
(S)-2,3,4,5-Tetrahydro-7-[[~[ 1 -(2-hydroxyethyl)ben7.imkl:~701-2-yl3methyl]amino]carbonyl]-4-methyl-3-oxo-lH-1,4-benzodiazepine-2-acetic acid;
(+)-2,3,4,5-Tetrahydro-7-[[[(4-aza-5-methylben~.i...irl~7.ol-2-yl)methyl]arnino]carbonyl]-4-(2-methoxyethyl)-3-oxo- 1 H- 1 ,4-benzodiazepine-2-25 acetic acid;
(S)-2,3,4,5-Tetrahydro-7-[[[(4-aminoben7.imill~701-2-yl)methyl]methylarnino]carbonyl~-4-methyl-3-oxo-lH-1,4-berl7.o~ 7Ppine-2-acetic acid;
Ethyl (S)-2,3,4,5-tetrahydro-7-[[~(ben7.imid~7.nl-2-30 yl)methyl]methylamino]carbonyl]~-methyl-3-oxo-lH-1,4-ben7.o~ 7.P.pine-2-acetate;
(S~-2,3,4,5-Tetrahydro-7-~[[(benzimidazol-2-yl)methyl]methylamino]carbonyl]-4-methyl-3-oxo- 1 H- 1 ,4-benzodiazepine-2-acetic acid, ~(2,2-dimethyl-2-methoxyacetyl)oxy]methyl ester;
2,3,4,5-Tetrahydro-7-[[[( l R)-(b~ 701-2-yl)ethyl]methylarnino]carbonyl]-4-methyl-3-oxo- 1 H- 1 ,4-benzodiazepine-(2S)-acetic acid;
(_)-N-t2-(Aminomethyl)-4-[[[(4-aza-5-methylben7.imi~1~7.(31-2-yl)methyl]methylamino]carbonyl]phenyl]aspartic acid;
(+)-2,3,4,5-Tetrahydro-4-methyl-3-oxo-7-[[[(phen~nthrimi~1~7.ol-2-yl)methyl]amino~carbonyl]-1H-1,4-benzodiazepine-2-acetic acid;
(+)-2,3,4,5-Tetrahydro-7-[3-(benzimidazol-2-yl)phenyl]-4-methyl-3-oxo- lH-1,4-ben70di~7~pine-2-acetic acid;
- (+)-4-[4-[[[(Ben7imid~701-2-yl)methyl]methylamino]carbonyl]phenyl]-3-(dimethylaminocarbonyl)butanoic acid;
(S)-2,3,4,5-Tetrahydro-7-[[[(ben7.imiclz-7nl-2-yl)methyl]amino]carbonyl]-3-oxo-4-[2-(pyrid-3-yl)ethyl]-lH-1,4-bP.n70~ 7~pine-2-acetate;
(S)-2,3,4,5-Tetrahydro-7-[[[(4-aza-5-methylben7imi~1~701-2-yl)methyl]methylamino]carbonyl]-3-oxo- 1 H- 1 ,4-benzodiazepine-2-acetic acid;
(+)-2,3,4,5-Tetrahydro-7-[[N-[(ben7imi-1~7.- 1-2-yl)methyl]-N-[[4-(2-carboxybenzoyl)amino]butyl]amino]carbonyl]-3-oxo-4-(2-phenylethyl)- 1~-1,4-benzodiazepine-2-acetic acid;
(+)-2,3,4,5-Tetrahydro-7-[[N-[(ben7.imi-1~7.ol-2-yl)methyl3-N-[[4-(4-azido-2-hydroxybenzoyl)amino]butyl]amino]carbonyl]-3-oxo-4-(2-phenylethyl)- 1 H- 1,4-benzodiazepine-2-acetic acid;
Ethyl (S)-2,3,4,5-tetrahydro-7-[[[(4-aza-5-methylbe~7imid~701-2-yl)methyl]amino~carbonyl]-4-methyl-3-oxo- I H- 1 ,4-benzodiazepine-2-acetate;
2,3,4,5-Tetrahydro-7-~[N-[(ben7.imil1~7rl-2-yl)methyl]-N-[[[(+)-biotinoyl]amino]butyl]amino]carbonyl]-3-oxo4-(2-phenylethyl)- 1 H- 1,4-benzodiazepine-(2RS)-acetic acid;
W O 97/24119 PCTnUS96/20748 2,3,4,5-Tetrahydro-7-[[[(lS)-(ben,.i...i-1~7.Ql-2-yl)ethyl]methylamino]carbonyl]-4-methyl-3-oxo-lH-1 ,4-benzodiazepine-(2S)-acetic acld;
(S)-2,3,4,5-Tetrahydro-7-[[[(imi~1~7.o( 1 ,2a)pyrid-2-S yl)methyl]methylamino]carbonyl] -4-methyl-3-oxo- 1 H- 1 ,4-bçn7.0rli ~7epine-2-acetic acid;
(S)-2,3,4,5-Tetrahydro-7-rt[(benz; ., .i~7.ol-2-yl)methyl]amino~carbonyl]-3-oxo-lH-1,4-berl7.orli:~7çpine-2-acetic acid;
(+)-5-[[2,3,4,5-Tetrahydro-7-~[[(benzimidazol-2-yl)methyl]amino]calbollyl]-3-oxo-4-(2-phenylethyl)- 1 H- 1 ,4-benzodiazepin-2-yl]methyl]tetra_ole;
(S)-2,3,4,5-Tetrahydro-7-[[[(4-aza-5-methylbel.~,i..licl~7.ol-2-yl)methyl]amino]carbonyl]-3-oxo- 1 H- 1 ,4-benzodiazepine-2-acetic acid;
(+)-2,3,4,5-Tetrahydro-7-[3-(ben7.imicl~7ol-2-yl)propyl]-4-methyl-3-oxo-lH-1,4-benzodiazepine-2-acetic acid;
(+)-2,3,4,5-Tetrahydro-7-[[N-[(benzill~dazol-2-yl)methyl]-N-(4-aminobutyl)amino]carbonyl] -3-oxo-4-(2-phenylethyl)- 1 H- 1 ,4-benzodiazepine-2-acetic acid;
(+)-2,3,4,5-Tetrahydro-7-[[[(ben7imi~ 7.ol-2-yl)methyl]amino]carbonyl]-4-methyl-3-oxo- lH- 1 ,4-benzodiazepine-2-(N-hydroxy)aret~mide;
Ethyl (+)-3-[[[2-(Ben7.imi~7.~1-2-yl)ethyl]amino]succinoyl]amino-4-pentynoate;
(+)-3-[[[2-(Bçn7.imi(1~7.ol-2-yl)ethyl~amino]succinoyl]amino4-pentynoic acid;
(+)-2,3 ,4,5-Tetrahydro-7-[[N-[(benzimidazol-2-yl)methyl] -N-[[4-(4-azido-3-iodo-2-hydroxybenzoyl)amino]butyl] amino]carbonyl] -3 -oxo-4-(2-phenylet'nyl)- 1 H-1,4-benzodiazepine-2-acetic acid;
2,3,4,5-Tetrahyciro-7-~[[(iS)-(ben7.imi~l~7.Qi-2-yi)ethyi]amino3carbonyl]-4-methyl-3-oxo- 1 H- 1 ,4-benzodiazepine-(2S)-acetic acid;
2,3,4,5-Tetrahydro-7-[[[( 1 R)-(benzimidazol-2-yl)ethyl]amino]carbonyl3-4-methyl-3-oxo-lH-1,4-benzodiazepine-(2S)-acetic acid; and .
w o 97n4119 PCTrUS96/20748 (+)-7-[[[(4,5-Dimethyl-lH-imi~1~7~1-2-yl)methyl]methyla~mino]carbonyl~-2,3,4,5-tetrahydro-4-methyl-3-oxo- lH- 1 ,4-benzodiazepine-2-acetic acid;
or pharmaceutically acceptable salts thereof.
The most preferred fibrinogen receptor antagonist template is of the sub-S formula (VIa), wherein CR2R2 is CHCH2CO2H, CR3R3 is C=O, and CRsRs isCH2. Vitronectin fibrinogen receptor antagonism is particularly pronounced when the A-W- substituent is attached to the 7-position of the 3-oxo-2,3,4,5-tetrahydro-lH- 1 ,4-benzodiazepine ring system.
In the formula below the definitions for the ~.ub~.LiLuents are as defined in 10 ft rm~ (I)-(V) and (XX)-(XXI), unless specified otherwise.
Another embodiment of a preferred fibrinogen receptor template A is represented by the 1,4-benzodiazepine 2,5-dione of sub-formula (VII);
~Y
R (VII) 1 5 wherein:
Y is H, Cl 4alkyl, Cl 4alkoxy, Cl 4alkoxycarbonyl, F, Cl, Br, I, CF3, oRf, S(O)kRf, CORf, NO2, N(Rf)2, CO(NRf)2, CH2N(Rf)2, methylenedioxy, CN, CO2Rf, OC(O)Rf, or NHC(O)Rf; and Rh is (CH2)qC02R
The plepal~tion and the use of tnis sub-structure in ~l~ing fibrinogen receptor antagonists of this sub-formula is detailed in Bondinell, et al., WO
93/OOO9S published January 7, 1993 and Blackburn, et al., WO 93/08174, publishedApril 29, 1993.
..
W O 97/24119 PCT~US96/20748 Table I, below, sl~mm~ries other ~l~felled fibrinogen receptor templates that are included within the scope of the present invention. Such templates are:
Table I
(VIII) Ais~R ~R22 J~R2~ j~R22 or / ~R
whGreill:
R21 and R22 independently are H or -Z-CO2Rfor Z-CON~Rf)2 with the proviso that one of Al or A2 is -ZCO2Rf or Z-CON(Rf)2;
Zis-CH2-,~O(CH2)q~~~NRf(CH2)q~~~S(CH2)q~-CH2CH2-,-CH(CH3)CH2-, -(CH2)3-, -CH=CH-, -C(CH3)=CH-, CH2-CH=CH- or CH=CHCH2; and Y is H, Cl 4alkyl, Cl 4alkoxy, Cl 4alkoxycarbonyl, F, Cl, Br, I, CF3, oRf, S(O)kRf, CORf, NO2, N(Rf)2, CO(NRf)2, CH2N(Rf)2, methylenedioxy or Z-CORf, in Alig, et al., EP 0 381 033, published August 8, 1990.
(IX) R6SO2NH CO2R~
20 wherein:
R6 is aryl, Cl lOalkyl, C3 6cycloalkyl, C4 1oaraLkyl, Cl loalkoxyalkyl, Cl loalkaryl, Cl loalkylthio~lkyl, Cl loaLkoxythioalkyl, Cl loalkylarnino, C4 1 oaralkylarnino, C 1-1 oalkanoylarnino, C4_ 1 oaralkanoylarnino, C 1-1 oaL~anoyl, C4 10aralkanoyl, or Cl locarboxyaLkyl; and S Y is H, Cl 4alkyl, Cl 4aLlcoxy, Cl 4alkoxycarbonyl, F, Cl, Br, I, CF3, ORf, S(o)kRf, CORf, NO2, N(Rf)2, Co(NRf)2, CH2N(Rf)2, methylenedioxy, CN, CO2Rf, OC(o)Rf, or NHC(O)Rf, in Egbertson, et al., EP 0 478 328, published April 1, 1992.
(X) --M~\M2~G~ CHCO2Rf wherein:
Ml is CH or N;
M2 is CH or N, with the proviso that when Ml is CH, M2 is N; and G' is N or N~3R", in Eldred, et al., EP 0542 363, published May 19, 1993.
. (XI) --M~\M2{><0H
--/ CH2CO2R~
wherein:
Ml is CH or N; and M2 is CH or N, with the proviso that when Ml is CH, M2 is N, in Porter, et al., EP 0 537 380, published April 21, 1993.
I
W O 97t24119 PCTrUS96/20748 ~XIIl ~3 . M ~ Rh wherein:
M1 isCHorN;
S Y is H, Cl 4alkyl, Cl 4alkoxy, Cl 4alkoxycarbonyl, F, Cl, Br, I, CF3, ORf, S(O)kRf, CORf, NO2, N(Rf)2, CO(NRf)2, CH2N(Rf)2, methylenedioxy, CN, Co2Rf, OC(O)Rf, or NHC(o)Rf;
D3 is CH2 or C=O; and Rh is (cH2)qco2Rf7 10in Klinnick, et al., EP 0 635,492, published January 25, l99S.
(XIII) \B~ N
wherein:
Y is H, C~ 4alkyl, C1 4alkoxy, Cl 4alkoxycarbonyl, F, Cl, Br, I, CF3, oRf, lS S(o)kRf, CORf, NO2, N(Rf)2, Co(NRf)2, CH2N(Rf)2, methylenedioxy, CN, CO2Rf, OC(O)Rf, or NHC(O)Rf;
Rh is ~CH2)nCO2Rf; and B is HsC)~ , ~N H3C ~
- in Blackburn, et al., WO 9S/04057, published February 9, 1995.
~V) W O 97/24119 PCT~US96/20748 ,~
N~ L -C02R9 O
wherein:
L* is -C(O)NRg-(CH2)-, ~C(O)~(CH2)q~~ NRg-(CH2)q-, ~O~(CH2)q~~ or S(O)k-(CH2)q~~
in Hartman, et al., EP 0 540 331, published May 5, 1993.
~V) N N--CH2 Co2R9 CO~o in Sugihara, et al., EP 0 529,858, published March 3, 1993.
(XVI~
~~
~ "'--CO R~
wherein:
Y is H, Cl 4alkyl, Cl 4alkoxy, Cl 4alkoxycarbonyl, F, Cl, Br, I, CF3, ORf, S(o)kRf, CoRf1, N02, N(Rf)2, Co(NRf)2, CH2N(Rf)2, methylenedioxy, CN, CO2Rf, Oc(o)Rf~ or NHC(O)Rf, in ~imm~i~b~h, et al., EP 0 483 667, published May 6, 1992.
(XVII) ~(CH2)oco2Ro N
in Linz, et al., LP 0 567 968, published November 3, 1993.
(XVIII) CA 0224l633 l998-06-26 W ~ 97/2~119 PCTAUS96/20748 Z"' Z~
~<Co2R9 wherein:
Rd is Het-C0 6alkyl; and Z", Z"' independently are hydrogen, Cl 4alkyl, halo, ORf, CN, S(o)kRf, 5 Co2Rf, or OH, in Bovy, et al., EP 0 539 343, published April 28, 1993.
Compounds of this invention comprising specific fibrinogen templates are named in the examples. These examples illustrate the invention, but do not limit the 10 scope of the invention.
The above descriptions of fibrinogen receptor templates for use in the present invention were taken from pending published patent applications. Reference should be made to such patent applications for their full disclosures, including the methods of plc~ali~lg said templzlt~s and specific compounds using said templates, the entire 15 disclosure of such patent applications being incol~oldl~d herein by reference.
Table II, below, describes other fibrinogen receptor antagonists, whose core structures would be useful in carrying out the instant invention. Reference should be made to the patent applications and other publications for their full disclosures, 20 including the methods of preparing said tt-mpl~t~s and specific compounds using said templates, the entire disclosure of the noted patent applications and otherpublications being incorporated herein by reference. Since it is contemplated that any fibrinogen receptor antagonist that is linked to an optionally fused nitrogen-cont~ining five-m.-.mhP.red ring will possess the novel utility described herein, the 25 list below does not limit the scope of the present invention.
WO 97124119 PCT~US96/20748 Table II
Adir et Col~r:-~nie FR 928004, June 30, 1992, Fauchere, J. L., et al.
EP 0578535, June 29, 1993, Fauchere, J-L, et al.: Describes X-RGDW-OH analogs, where X contains a cationic arnine.
CA 2128560, Jan. 24, 1995, Godfroid, J-J, et al., substituted pipel~ines.
Asahi Breweries, Ltd.
JP 05239030, Sep. 17, 1993, aminomethyltetrahydroisoquinolines.
Asahi Glass WO 90/02751, Ohba, M. et al.: Sept. 8, 1989: Describes cyclic RGD-con~ining peptides.
WO 90/115950, Mar. 22, 1990, Ohba, M., et al.
EP 0406428, 1/9/91: Describes cyclic RGD-cont:~ining peptides WO 92/09627, Isoai, A. et al.: Nov. 29, 1991: Describes cyclic RGD-cont~ining peptides.
~s~c~PIls AG
DE 4207254, (Der 93-289298/37) Mar. 7, 1992, Zoller, G., et al.: Describes guanidinopropyl-4-oxo-2-thioimida_olidin-3-yl-Asp-X analogs EP 93904010, Feb. 24, 1993, Zoller, G., 4-oxo-2-Thioxoimi~1~7Olidine Derivatives.
EP 0565896, Mar. 18, 1993, Klinger, O, et al.: Describes guanidinoethylphenyloxyacetyl-Asp-X analogs.
EP 0566919, (Der 93-338002/43) Apr. 3, 1993, Zoller, G., et al.: Describes guanidinopropyl-4-oxo-2-thioimidazolidin-3-yl-Asp-X analogs.
EP 580008, (Der 94-027663/04) July 6, 1993, Zoller, G., et al.: Describes S-m-guanidinophenyl-2,4-dioxoimi(l~7.Qlidin-3yl)acetyl-Asp-Phg.
DE 224414, July 6, 1993, Zoller, G., et al.: Describes 5-m-~tl:~ni~inophenyl-2,4-dioxoimidazolidin-3yl)acetyl-Asp-Phg.
W O 97/24119 PCT~US96/20748 EP 584694, (Der 94-067259/09) Apr. 2, 1994, Zoller, G., et al.: Describes 5-m-guanidinophenyl-2,4-dioxoimidazolidin-3yl~acetyl-Asp-Phg .
" DE 4301747, (Der 94-235891/29) Jul. 28, 1994, Zoller, G., et al.: Describes 5-m-guanidinophenyl-2,4-dioxoimi(1~7O1idin-3yl)acetyl-Asp-Phg analogs.
DE 4308034, (Der 94-286666/36) Sept. 15, 1994, Klinger, O. et al.: Describes 5-m-guanidinophenyl-2,4-dioxoimi~i~7O1idin-3yl)acetyl-Asp-Phg analogs.
DE 4309867, Sept. 29, 1994, Klingler, O, et al.: Describes 5-m-guanidinophenyl-2,4-dioxoimi~ 7.olidin-3yl)acetyl-Asp-Phg.
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Ciba Geigy EP 0452210, (Der 91-305246/42) Apr, 5, 199Q, describes aminoalkanoyl-GDF
analogs.
EP 0452257, Mar. 26, 1991, Allen, M. C., et al.: Describes aminoalkanoylAsp-Phe analogs.
COR Ther~renti~c WO 90/15620, June 15, 1990: Describes cyclic RGD-cont~ining peptides.
EP 0477295, Apr. 1, 1992: Scarborough, R. M. et al.
WO 92/08472, May 29, 1992, Scarborough, R. M. et al.
WO 93/223356, April 27. 1993, Swift, R. L., et al.: Describes cyclic RGD-c--nt~inin~ peptides.
EP 0557442, Sept. 1, 1993, Scarborough, R. M., et al.
Scarborough, R. M.; Rose, J. W.; Hsu, M. A.; Phillips, D. R.; Fried, V. A.;
Campbell, A. M.; N--nni77i, L.; Charo, I. F., Barbourin, A GPIIb-IIIa-Specific Integrin Antagonist from the Venom of Sistrurus M. Barbouri, J.
BioL Chem., 266, 9359, 1991.
CA 0224l633 l998-06-26 W O97/24119 PCT~US96/20748 Daiichi Pharm Co Ltd.
JP 05078344-A, ~Der 93-140339/17) Mar. 30, 1993: Describes Bis-amidinoheter~cycles, eg. benzofurans.
S DuPont Merck WO 93/07170, Apr. lS, 1993: Describes cyclic-RGD-con~ining peptides.
WO 94/11398, May 26, 1994: Wells, G. J. et al. Describes cyclic RGD cont~ining peptides.
IL 109237, Ju1. 31, 1994.
WO 94/22909, (Der 94-333113/41) Oct. 13, 1994: DeGrado W. F., et al.
WO 94/22910, (Der 94-333114/41 Oct. 13, 1994: DeGrado W. F., et al. Prodrugs.
Vitronectin Receptor Antagonists FIELD OF THE INVENTION
This invention relates to pharm~eu~ically active compounds which inhibit the vitronectin receptor and are useful for the treatment o~ infl~mm~tion, cancer and cardiovascular disorders, such as atherosclerosis and restenosis, and ~ e:~e.s wherein bone resorption is a factor, such as osteoporosis.
BACKGROUND OF TH~ INVEI~TION
Integrins are a ~upelr~lllily of cell adhesion receptors, which are transmembrane glycoproteins expressed on a variety of cells. These cell surface adhesion receptors include gpIIb /IIIa, the fibrinogen receptor, and o~vM3, the 15 vitronectin receptor. The fibrinogen receptor gpIIb /IIIa is expressed on the platelet surface and it m~ tes platelet aggregation and the formation of a hemostatic clot at the site of a bleeding wound. Philips, et al., Blood., 1988, 71, 831. The vitronectin receptor av1~3 is expressed on a number of cells, including endothelial, smooth muscle, osteoclast, and tumor cells, and, thus, it has a variety of functions. The aV133 20 receptor expressed on the membrane of osteoclast cells mediates the bone resportion process and contributes to the development of osteoporosis. Ross, et al., J. Biol.
Chem., 1987, 262, 7703. The ocv~3 receptor expressed on human aortic smooth muscle cells ~tim~ tPs their migration into neointima, which leads to the formation of atherosclerosis and restenosis after angioplasty. Brown, et al., Cardiovascular 25 Res., 1994, 28, 1815. Additionally, a recent study has shown that a CCv~3 antagonist is able to proIrlote turnor regression by in.lu.~ing aEioptosi~. of angiogenic l~lood vessels. Brooks, et al., Cell, 1994, 79, 1157. Thus, agents that would block thevitronectin receptor would be useful in treating diseases m~ t~d by this receptor, such as osteoporosis, atherosclerosis, restenosis and cancer.
~ -- 1 --W O97/24119 PCT~US96/20748 The vitronectin receptor is known to bind to bone matrix proteins, such as osteopontin, bone sialoprotein and thrombospondin, which contain the tri-peptideArg-Gly-Asp (or RGD) motif. Thus, Horton, et al., Exp. Cell Res. lg91, 195, 368,disclose that RGD-cont:lining peptides and an anti-vitronectin receptor antibody(23C6~ inhibit dentine resorption and cell spreading by osteoclasts. In addition, Sato, et al., J. Cell Biol. 1990, 111, 1713 disclose that echicf~fin, a snake venom peptide which contains the RGD sequence, is a potent inhibitor of bone resorption in tissue culture, and inhibits attachment of osteoclasts to bone. Fisher, et al., Endocrinology 1993, 132, 1411, has further shown that echistatin inhibits bone resorption in vivo in the rat. BertoIini etal., J. Bone Min. ~es., 6, Sup. 1, S146, 252 have shown that cylco-S,S-N~C-acetyl-cy~leillyl-N~c methyl-argininyl-glycyl-aspartyl-penicillzlmin~ inhibits osteoclast ~tt:~hmt-nt to bone. EP 528 587 and 528 586 report substituted phenyl derivatives which inhibit osteoclast m~ tçA bone resorption.
Alig et al., EP 0 381 033, Hartman, et al., EP 0 540,334, Blackburn, et al., WO 93/08174, Bondinell, et al., WO 93/00095, Blackburn, et al. WO 95/04057, Egbertson, et al, EP 0 478 328, Sugihara, et al. EP 529,858, Porter, et al., EP 0 542 363, and Fisher, et al., EP 0 635 492 ~1icclc)se certain compounds that are useful for inhibiting the fibrinogen receptor. It has now been discovered that certain a~ iately substituted compounds are potent inhibitors of the vitronectin receptor.
In particular, it has been discovered that such compounds are more potent inhibitors of the vitronectin receptor than the fibrinogen receptor and such compounds contain a fibrinogen receptor antagonist template.
SUM[MARY OF THE INVENTION
This invention comprises compounds of the formula (I)-(V) and (XXI)-(XXII) as described hereinafter, which have pharmacological activity for the inhibition of the vitronection receptor and are useful in the treatment of infl~mm:-fion, cancer and cardiovascular disorders, such as atherosclerosis and restenosis, and ~lice~cçs wherein bone resorption is a factor, such as osteoporosis.
W O g7/24119 PCTrUS96120748 This invention is also a pharm~reutical composition comprising a compound according to formula (I)-(V) (XXI)-(XXII)and a ph~rm:~reutically carrier.
This invention is also a method of treating ~ e~es which are m~ t~d by " the vitronectin receptor. In a particular aspect, the compounds of this invention are 5 useful for treating atherosclerosis, restenosis, infl~mm~tion, cancer and diseases wherein bone resorption is a factor, such as osteoporosis.
DETAILI~D DESCRIPTION
This invention compri~es novel compounds which are more potent inhibitors 10 of the vitronectin receptor than the fibrinogen receptor. The co~ oullds of the instant invention comprise a fibrinogen receptor antagonist template that is linked to a nitrogen-cont~ining five-membered ring, which is optionally fused to an aromatic six-membered ring. The fibrinogen receptor antagonist template is substituted by an aliphatic substituent which contains an acidic moiety. It is preferred that about 15 fourteen intervening covalent bonds via the shortest intramolecular path will exist between the acidic group of the fibrinogen receptor antagonist template and the nitrogen of the optionally fused five-membered ring.
As used herein, the term "fibrinogen recel~lor antagonist template" means the core structure of a fibrinogen receptor antagonist, said core being substituted by an 20 acidic group and said core being linked to an organic group substituted with a basic nitrogen moiety. A fibrinogen receptor antagonist is an agent that inhibits the binding of fibrinogen to the platelet-bound fibrinogen receptor GPIIb-IIIa. It is an object of this invention that a fibrinogen receptor antagonist is converted to avitronectin receptor antagonist by replacing the organic group substituted with a 25 basic nitrogen moiety in a fibrinogen receptor antagonist with an optionally fused nitrogen-cont~ining five-membered ring, preferably an imidazole ring and, most preferably, a ben~imi~ ole ring.
W O 97/24119 PCTrUS96/20748 This invention conl~lises compounds of formula (I)~
W A Rb G
,~ /~ R' ~W--A
(I) or (Il) or (1ll) or R" Rc~N--W A
(IV) or (V) wherein:
W is - (CHRg)b-V'- or Ar;
A is a fibrinogen receptor antagonist template;
V' is CONR21 or NR2~CO;
GisNRe,SorO;
Rg is H, Cl 6alkyl, Het-CO 6alkyl, C3 7cycloalkyl-CO 6alkyl or Ar- CO 6alkyl;
R21 is Het-CO 6alkyl-U'-CI 6alkyl-~ C3 7cycloalkyl-CO 6alkyl-U'-CI 6alkyl-, or Ar-CO 6alkyl-U'-C I 6alkyl-;
U' is CONRf or NRfCO;
Rf is H, Cl 6alkyl or Ar-CI 6alkyl;
Re is H, Cl 6alkyl, Ar-CI 6alkyl, Het-CI 6alkyl, C3 7cycloalkyl-CI 6alkyl, (CH2)qOH or (CH2)kCO2Rg;
kisO, 1 or2;
q is 1 or 2;
bisO, 1 or2;~0 Rb and RC are independently sele.cte~l from H, Cl 6alkyl, Ar-CO 6alkyl, Het-CO 6alkyl, or C3 6cycloalkyl-CO 6alkyl, halogen, C1~3, ORf, S(O)kRf, CORf, NO2, N(Rf)2, Co(NRf)2, CH2N(Rf)2, or Rb and RC are joined together to form a five or six membered aromatic or non-aromatic carbocyclic or heterocyclic ring, optionally substituted by up to three substituents chosen W O 97/24119 PCT~US96/20748 from halogen, CF3, Cl 4alkyl, ORf~ S(O)kRf, CORf, CO2Rf OH, N02, N(Rf)2, CO(NRf)2, and CH2N(Rf)2, or methylenedioxy;
or a pharm~-~elltically acceptable salt thereof.
This invention also comprises compounds of formula (XXI)-(XXII):
I~I~ G
~ B A ~ /~ B A
~XXI) or (XXII) wherein:
B is a linking moiety of the form -(CHRg)a-U- (CHRg)b-V-;
A is a fibrinogen receptor antagonist template;
G is NRe, S or O;
Rg is H, Cl 6alkyl, Het-CO 6alkyl, C3 7cycloalkyl-C0 6alkyl or Ar- C0 6alkyl;
Rk is Rg, -C(O)Rg, or -C(O)ORf;
Ri is is H, Cl 6alkyl, Het-CO 6alkyl, C3 7cycloalkyl-C0 6alkyl, Ar- C0 6alkyl, Het-CO 6aL~cyl-U-C1 6alkyl-, C3 7cycloalkyl-CO 6alkyl-U'-Cl 6alkyl-, or Ar-CO 6alkyl-U'-CI 6alkyl- or Cl 6alkyl;
lS Rf is H, Cl 6alkyl or Ar-CI 6alkyl;
Re is H, Cl 6alkyl, Ar-CI 6alkyl, Het-CI 6alkyl, C3 7cycloalkyl-CI 6alkyl, (CH2)qOH or (CH2)kCO2Rg;
U, U' and V independently are absent or CO, CRg2, C(=CRg2), S(O)k, O, NRg, CRgORg, CRg(ORk)CRg2, CRg2CRg(ORk), C(O)CRg2, CRg2C(O), CON Ri N Ri CO OC(O), C(O)O, C(S)O, OC(S), C(S)NRg, NRgC(S), S(O)2NRg, NRgS(O)2, N=N, NRgNRg, NRgCRg2, NRgCRg2, CRg2O, OCRg2, C_C, CRg=CRg~ Ar or Het;
kisO, 1 or2;
~ qis I or2;
W O 97/24119 PCT~US96/20748 aisO, 1 or2;
bisO, 1 or2;
Rb and RC are independently selected from H, Cl 6alkyl, Ar-CO 6alkyl, Het-CO 6alkyl, or C3 6cycloalkyl-CO 6alkyl, halogen, CF3, oRf, S(o)kRf, CORf, N02, N(Rf)2, Co(NRf)2, CH2N(Rf)2, or Rb and RC are joined together to forrn a five or six membered aromatic or non-aromatic carbocyclic or ~ heterocyclic ring, optionally- substituted by up to three substituents chosen from halogen, CF3, Cl 4alkyl, ORf, S(O)kR~, CORf, C02Rf OH, N02, N(Rf)2, CO(NRf)2, and CH2N(Rf)2; or methylenedioxy;
10 or pharm~reuticSllly acceptable salts thereof.
Preferably, U' is CONRf or NRfCO.
Also included in this invention are pharm:3relltically acceptable addition salts, complexes or prodrugs of the compounds of this invention. Prodrugs are considered to be any covalently bonded carriers which release the active parent drug lS according to formula (I) in vivo. In cases wherein the compounds of this invention may have one or more chiral centers, unless specified, this invention includes each unique nonracernic compound which may be synth~o~i7ç~1 and resolved by conventional techniques. In cases in which compounds have unsaturated carbon-carbon double bonds, both the cis (Z) and trans (E) isomers are within the scope of 20 this invention. In cases wheleill compounds may exist in tautomeric forms, such as O OR' keto-enol tautomers, such as ~~ and b ~, and each tautomeric form is contemplated as being included within this invention whether existing in equilibrium or locked in one form by a~ iate substitution with R'.
The compounds of formula (I) - (V) and (XXI) - (XXII) inhibit the binding of vitronectin and other RGD-co..~ lg peptides to the vitronectin (ocv1~3) receptor.
Inhibition of the vitronectin receptor on osteoclasts inhibits osteoclastic boneresorption and is useful in the tre~tm~nt of ~ e~sç~ wherein bone resorption is associated with pathology, such as osteoporosis. Additionally, since the compounds W O 97/24119 PCTrUS96120748 of the instant invention inhibit vitronectin receptors on a number of different types of cells, said compounds would be useful in the tre~tm~n~ of infl~mm~tion and cardiovascular diseases, such as atherosclerosis and restenosis, and would be useful as anti-m~t~cta~ir and ~ntitllmt~r agents.
In a particuar embodiment, the compounds of this invention are of the formula (II), wherein Rb and Rc are joined to form an aromatic ring ct-nt~ining up to two nitrogen atoms. In a preferred embodiment Rb and Rc are joined to form an optionally substituted phenyl ring according to formula (IIa):
RY ~C /~ W-A
R (IIa) wherein G is N-R, S, CH or 0.
L N-CO
Suitably W is -(CHRg)aNRiCO- or ~~ , or, when G is CH, W is -CH2CH2NRiCO-wherein Ri is a methylene group :lt~hl-cl to G.
Preferably W is -CHRgNRiC0-.
Suitably Ri is H, Cl 6alkyl, C3 7cycloalkyl, Ar or Cl 6alkyl substituted by one to three groups chosen from halogen, CN, NRg2, ORg, SRg, C02Rg, and CON(Rg)2, Ar, Het or C3 7cycloalkyl. In particular, Ri is H, methyl, butyl, cyanomethyl, carboxymethyl, phenylethyl or ben7imi~ 701ylmethyl.
Suitably RX, RY and RZ are independently chosen from Cl 6alkyl, methoxy, 20 nitro, trifluoromethyl, fluoro, chloro, amino or Rx and RY are adjacent to one another and are joined to form a methylenedioxy group.
Preferably G is NRe.
Suitably Re is H, Cl 4alkyl, Ar, Het or Cl 4alkyl substituted by Ar or Het.
More suitably, Re is H, methyl or ben7.imi~1~7olylmethyl.
W O 97/24119 PCT~US96/20748 In another speeific embodiment, Rb and Rc form a six membered aromatic ring eon~ining one or two nitrogen atoms according to formulas (IIb-d):
N R~ \~W A ~ \~W A
(lIb) (IIc) (IId) 5 wherein G, RX and RY are as above for formula (IIa).
Specifically, the compounds of this invention are comprised of a nitrogen-cont~ining optionally fused five-m~mhered ring, a linking group W, and a fibrinogen reeeptor antagonist template A. In partieular, the fibrinogen reeeptor antagonist template A is as defmed in Bondinell, et al., WO 93/00095, publich~ocl January 7, 1993, of the sub-formula (VI):
D A'--A (VI) Al to As form an ~reeccihle ~ub~LiLuLed seven-membered ring, which may be saturated or unsaturated, optionally eontaining up to two heteroatoms ehosen from 15 the group of O, S and N wlle~ S and N may be optionally oxi~
Dl to D4 form an aeeessible substituted six membered ring, optionally cont~ining up to two nitrogen atoms;
R is at least one substituent ehosen from the group of R7, or Q-Cl 4alkyl, Q-C2 4alkenyl, Q-C24alkynyl, optionally substituted by one or more of =O, Rl 1 or 20 R7;
R* is H, Q-Cl 6alkyl, Q-C1 6oxoalkyl, Q-C2 6alkenyl, Q-C3 40xoalkenyl, Q-C3 40xoalkynyl, Q-C24alkynyl, C3 6eyeloalkyl, Ar or Het, optionally substituted by one or more of Rl l;
Q is H, C3 6eyeloalkyl, Het or Ar;
CA 0224l633 l998-06-26 PCTrUS96/20748 R7 is -COR8, -COCR'2R9, -C(S)R8, -S(O)mOR', -S(O)mNR'R", -PO(OR'), -PO(OR')2, -B(OR')2, -NO2 and Tet;
R8 is -OR', -NR'R", -NR'SO2R', -NR'OR', -OCR'2C(O)OR', -OCR'2OC(O)-R', -OCR'2C(O)NR'2, CF3 or AAl;
R9 is -OR', -CN, -S(O)rR', S(O)mNR'2, -C(O)R' C(O)NR'2 or -CO2R';
Rl I is H, halo, -OR12, -CN, -NR'RI2, -NO2, -CF3, CF3S(O)r, -CO2R', -CONR 2, Q-C0 6alkyl-, Q-Cl 6oxoalkyl-, Q-C2 6alkenyl-, Q-C2 6alkynyl-, Q-Co 6alkyloxy-, Q-C0 6alkylamino- or Q-C0 6alkyl-S(O)~;
R12 is R', -C(O)R', -C(O)NR'2, -C(O)OR15, -S(O)mR' or S(O)mNR'2;
Rl3 is R', -CF3, -SR', or -OR';
Rl4 is R', C(O)R', CN, NO2, SO2R' or C(O)oRl5;
Rls is H, Cl 6alkyl or Ar-Co 4alkyl;
R' is H, Cl 6alkyl, C3 7cycloalkyl-Co 4alkyl or Ar-Co 4alkyl;
R" is R', -C(O)R' or-C(O)OR15;
R"' is R" or AA2;
AAl is an amino acid attached through its amino group and having its carboxyl group optionally protected, and AA2 is an amino acid attached through its carboxyl group, and having its amino group optionally protected;
mis 1 or2;
nisOto3;
pisOor l;and tisOto2;or ph~ celltic~lly acceptable salts thereof.
With reference to formula (VI), suitably, Al is CRlRl, CRl, NRl, N, O or S(O)x;
A2 iS CR2R2~ CR2, NR2;
A3 is CR3R3, CR3, NR3, N, O or S(O)x;
g CA 02241633 l99X-06-26 W O 97/24119 PCTrUS96120748 A4 is CR4R4, CR4, NR4, or N;
A5 is CRsRs, CRs, NR5, N, O or S(O)x;
Dl-D4 are CRIl~ CR6 orN;
Rl and Rl are R* or R, or together are =O;
R2 and R2 are R*, R or =O;
R3andR3 areR*,Ror=O;
R4 and R4 are R*, R or =O; .
Rs and R5 are R*, R or =O; and xisOto2.
More suitably, Al is CRlRl, CR1, NRl, N, O or S; A2 is CR2R2, NR2 or CR2; A3 is CR3R3; A4 is CR4R4, CR4, NR4, or N; AS is CR5Rs, CR5, NR5, N, O;
Dl - D4 are CH; R2 or R4 are R; R3,R3 and R5,R5 are =0 or R*,H.
Preferably, Al is CHRl, CRl, NR", N or S; A2 is CR2 or CR2R2; A3 is CR3R3; A4 is CR4R4 or NR4; As is CR5Rs, and Dl - D4 are CH.
In one embodiment, Al is CRl, A2 is CR2, A3 is C=O, A4 is NR4 and As are CHRs.
In another embodiment, Al is NRl, A2 is CHCR2, A3 is CR3R3, A4 is NR4, and A5 are C=O.
In yet another embodiment, Al and A4 are C=O, A2 is NR2, A3 is CHR3 and 20 AS is NRs.
In a preferred embodiment, Al is NRI, A2 is CHR2, A3 is C--O, A4 is NR' and A5 is CHRs.
Representative sub-form~ of (VI) are given by each of formulas (VIa)-(VIi) below:
,R-I ~5N,FI,~R3, R R2 N R2 R?~R ' (VIa) (VIb) (VIc) W O 97/24119 PCT~US96120748 , R~
(VId) (VIe) (VIf) {~ ~ )~ R3 Rl R1 2 , R2 or R1 R1' (VIg) (VIh) (VIi) Specific embodiments of this invention wherein the fibrinogen receptor antagonist template A is of the sub-formula (VI) are named in the Examples.
Preferred compounds of this invention are:
5-[[[(Benzimidazol-2-yl)methyl]methylamino]carbonyl]-lH-ben7.imicl:~701e-2-aminoacetic acid;
(+)-2,3,4,5-Tetrahydro-7-[[[(ben7imi~i~7.ol-2-yl)methyl}-methylamino~carbonyl]-4-(3 ,3-dimethylbutyl)-3-oxo- 1 H- 1 ,4-benzodiazepine-2-acetic acid;
(S)-2,3,4,5-Tetrahydro-4-methyl-3-oxo-7-[[[(5-trifluoromethylb~n7.imi(1~7.ol-2-yl)methyl]methylamino]carbonyl]- 1 H- 1 ,4-benzodiazepine-2-acetic acid;
(S)-2,3,4,5-Tetrahydro-7-[[[(4,7-dimethoxyben7.imi~1~7.ol-2-yl)methyl]methylamino~carbonyl]-4-methyl-3-oxo- 1 H- 1 ,4-benzodiazepine-2-acetic acid;
(+)-2,3,4,5-Tetrahydro-7-[[[(benzimidazol-2-yl)methyl]amino]carbonyl]-4-(3 ,3-dimethylbutyl)-3-oxo- 1 H- 1 ,4-benzodiazepine-2-acetic acid;
~. (S)-2,3,4,5-Tetrahydro-7-[[[(4-methylb~n7.imicl~7.ol-2-yl)methyl~methylamino]carbonyl]-4-methyl-3-oxo- 1 H- 1 ,4-benzodiazepine-2-acetic acid;
S3-2,3,4,5-Tetrahydro-7-[[N-[(ben7.imill~7nl-2-yl)methyl]-N-(4-anninobutyl3amino]carbonyl]-4-methyl-3-oxo-lH-1,4-benzodiazepi.le-2-acetic acid;(S)-2,3,4,5-Tetrahydro-7-[[N-(ben ~.i ., ~idzl701-2-yl)methyl-N-(2-cyanomethyl)amino]carbonyl]-4-methyl-3-oxo-lH-1,4-b~.n70~ Ppine-2-acetic 5 acid;
(S)-2,3,4,5-Tetrahydro-7-[[[(be~7.imi~1~7.ol-2-yl)methyl]amino~carbonyl]-3-oxo-4-(4-phth~limiclobutyl)-lH-l~4-benzodiazepine-2-acetic acid;
4-[[r3-(Ren7.imitl~701-2-yl)propyl]amino]carbonyl]piperidine-1-acetic acid;
4-[[~3-(Ben7.imi~1~701-2-yl)propyl]amino]carbonyl]phenylacetic acid;
(S)-2,3,4,5-Tetrahydro-7-[[[(4-aza-5,7-dimethylben7.imi~ 7-1-2-yl)methyl3methylamino~carbonyl]-4-methyl-3 -oxo- 1 H- 1 ,4-benzodiazepine-2-acetic acid;
(~t)-2,3,4,5-Tetrahydro-7-[[[(bel~7.i,..i~1~7.ol-2-yl)methyl3methylamino~-carbonyl}-4-[2-(3 ,4-methylenedioxyphenyl)ethyl]-3-oxo- 1 H- 1 ,4-benzodiazepine-2-acetic acid;
(~)-2,3 ,4,5-Tetrahydro-7-[[[(ben7.imidz-701-2-yl)methyllamino]carbonyl]-4-(2-methoxyethyl)-3 -oxo- 1 H- 1 ,4-benzodiazepine-2-acetic acid;
(S)-2,3,4,5-Tetrahydro-7-[[[(bt~n7.imi~1~7.ol-2-yl)methyl]methylamino]-carbonyl]-4-methyl-3-oxo- 1H- 1 ,4-benzodiazepine-2-~f et~mi~le;
(+)-2,3,4,5-Tetrahydro-7-[[[[1-[(ben7.imi~1~7.ol-2-yl)methyl]ben7.imi(1~7.ol-2-yl]methyl]amino]carbonyl]4-methyl-3-oxo-lH-1,4-ben7.orliz~7epine-2-acetic acid;
(S)-2,3,4,5-Tetrahydro-7-[[[(be.n7.imi-1~7.~-1-2-yl)methyl]methylamino]carbonyl~-3 -oxo- 1 H- 1 ,4-benzodiazepine-2-acetic acid;
(S)-2,3,4,5-Tetrahydro-7-[[bis[(benzimidazol-2-yl)methyl]amino]carbonyl]-4-methyl-3-oxo- lH-1 ,4-benzodiazepine-2-acetic acid;
(+)-2,3,4,5-Tetrahydro-7-r[[(4-azaben7.imit1~701-2-yl)methyl~methylamino]carbonyl]-4-(3 ,3 -dimethylbutyl)-3-oxo- 1 H- 1,~
benzodiazepine-2-acetic acid;
(i~)-2,3,4,5-Tetrahydro-7-[[[(ben7.imi<1~7.Ql-2-yl)methyl]methylamino]carbonyl]-3-oxo-4-(2,2,2-trifluoroethyl)-lH-1,4-benzodiazepine-2-acetic acid;
CA 0224l633 l998-06-26 W O 97/24119 PCT~US96/20748 (~)-2,3,4,5-Tetrahydro-7-t[2-(ben7.imid~7.ol-2-yl)acetyl~amino]-S-oxo-4-(2-phenylethyl)- 1 H- 1 ,4-benzodiazepine-2-acetic acid;
, (+)-2,3,4,5-Tetrahydro-7-[[[(ben7.imi-1~7.ol-2-yl)methyl]amino]carbonyl]-3-oxo-4-(2,2,2-trifluoroethyl)- lH- 1 ,4-benzodiazepine-2-acetic acid;
" S (S)-2,3,4,5-Tetrahydro-7-~[[(5,6-difluoroberl7.imifl:~7.ol-2-yl)methyl]methylamino]carbonyl]-4-methyl-3-oxo- lH- 1 ,4-benzodiazepine-2-aceticacid;
(+)-2,3,4,5-Tetrahydro-7-[[bis[(ben7.imicl~7.ol-2-yl)methyl]amino]carbonyl]-3-oxo-4-(2-phenylethyl)- lH- 1 ,4-ben70~ 7.epine-2-acetic acid;
(S)-2,3,4,5-Tetrahydro-7-[[[~4-aza-5-methylben7imid:~7.ol-2-yl)methyl]arnino]carbonyl]-4-methyl-3-oxo- 1 H- 1 ,4-benzodiazepine-2-acetic acid;
(S)-2,3,4,5-Tetrahydro-4-methyl-7-[[[(4-nitroben7.imi(1~7.ol-2-yl)methyl]methylaminolcarbonyl]-3-oxo- lH- 1 ,4-benzodiazepine-2-acetic acid;
(+)-2,3,4,5-Tetrahydro-7-[[[(4-aza-S-methylben7.imi~ 7.ol-2-15 yl)methyl]amino]carbQnyl]-3-QxQ-4-(2,2,2-trifluoroethyl)- lH- 1 ,4-benzodiazepine-2-acetic acid;
(+)-4-~4-[[L( iH-Ben7.imidi~7.oi-2-yi)melhyljmethylamino]ccubonyl]phenyl]=3=
phenylbutanoic acid;
(+)-3-[[[4-(4-Azabe.,~ 7.Ql-2-yl)butanoyl]glycyl]amino]-4-pentynoic 20 acid;
(S)-2,3,4,5-Tetrahydro-7-[[~[ 1 -(2-hydroxyethyl)ben7.imkl:~701-2-yl3methyl]amino]carbonyl]-4-methyl-3-oxo-lH-1,4-benzodiazepine-2-acetic acid;
(+)-2,3,4,5-Tetrahydro-7-[[[(4-aza-5-methylben~.i...irl~7.ol-2-yl)methyl]arnino]carbonyl]-4-(2-methoxyethyl)-3-oxo- 1 H- 1 ,4-benzodiazepine-2-25 acetic acid;
(S)-2,3,4,5-Tetrahydro-7-[[[(4-aminoben7.imill~701-2-yl)methyl]methylarnino]carbonyl~-4-methyl-3-oxo-lH-1,4-berl7.o~ 7Ppine-2-acetic acid;
Ethyl (S)-2,3,4,5-tetrahydro-7-[[~(ben7.imid~7.nl-2-30 yl)methyl]methylamino]carbonyl]~-methyl-3-oxo-lH-1,4-ben7.o~ 7.P.pine-2-acetate;
(S~-2,3,4,5-Tetrahydro-7-~[[(benzimidazol-2-yl)methyl]methylamino]carbonyl]-4-methyl-3-oxo- 1 H- 1 ,4-benzodiazepine-2-acetic acid, ~(2,2-dimethyl-2-methoxyacetyl)oxy]methyl ester;
2,3,4,5-Tetrahydro-7-[[[( l R)-(b~ 701-2-yl)ethyl]methylarnino]carbonyl]-4-methyl-3-oxo- 1 H- 1 ,4-benzodiazepine-(2S)-acetic acid;
(_)-N-t2-(Aminomethyl)-4-[[[(4-aza-5-methylben7.imi~1~7.(31-2-yl)methyl]methylamino]carbonyl]phenyl]aspartic acid;
(+)-2,3,4,5-Tetrahydro-4-methyl-3-oxo-7-[[[(phen~nthrimi~1~7.ol-2-yl)methyl]amino~carbonyl]-1H-1,4-benzodiazepine-2-acetic acid;
(+)-2,3,4,5-Tetrahydro-7-[3-(benzimidazol-2-yl)phenyl]-4-methyl-3-oxo- lH-1,4-ben70di~7~pine-2-acetic acid;
- (+)-4-[4-[[[(Ben7imid~701-2-yl)methyl]methylamino]carbonyl]phenyl]-3-(dimethylaminocarbonyl)butanoic acid;
(S)-2,3,4,5-Tetrahydro-7-[[[(ben7.imiclz-7nl-2-yl)methyl]amino]carbonyl]-3-oxo-4-[2-(pyrid-3-yl)ethyl]-lH-1,4-bP.n70~ 7~pine-2-acetate;
(S)-2,3,4,5-Tetrahydro-7-[[[(4-aza-5-methylben7imi~1~701-2-yl)methyl]methylamino]carbonyl]-3-oxo- 1 H- 1 ,4-benzodiazepine-2-acetic acid;
(+)-2,3,4,5-Tetrahydro-7-[[N-[(ben7imi-1~7.- 1-2-yl)methyl]-N-[[4-(2-carboxybenzoyl)amino]butyl]amino]carbonyl]-3-oxo-4-(2-phenylethyl)- 1~-1,4-benzodiazepine-2-acetic acid;
(+)-2,3,4,5-Tetrahydro-7-[[N-[(ben7.imi-1~7.ol-2-yl)methyl3-N-[[4-(4-azido-2-hydroxybenzoyl)amino]butyl]amino]carbonyl]-3-oxo-4-(2-phenylethyl)- 1 H- 1,4-benzodiazepine-2-acetic acid;
Ethyl (S)-2,3,4,5-tetrahydro-7-[[[(4-aza-5-methylbe~7imid~701-2-yl)methyl]amino~carbonyl]-4-methyl-3-oxo- I H- 1 ,4-benzodiazepine-2-acetate;
2,3,4,5-Tetrahydro-7-~[N-[(ben7.imil1~7rl-2-yl)methyl]-N-[[[(+)-biotinoyl]amino]butyl]amino]carbonyl]-3-oxo4-(2-phenylethyl)- 1 H- 1,4-benzodiazepine-(2RS)-acetic acid;
W O 97/24119 PCTnUS96/20748 2,3,4,5-Tetrahydro-7-[[[(lS)-(ben,.i...i-1~7.Ql-2-yl)ethyl]methylamino]carbonyl]-4-methyl-3-oxo-lH-1 ,4-benzodiazepine-(2S)-acetic acld;
(S)-2,3,4,5-Tetrahydro-7-[[[(imi~1~7.o( 1 ,2a)pyrid-2-S yl)methyl]methylamino]carbonyl] -4-methyl-3-oxo- 1 H- 1 ,4-bçn7.0rli ~7epine-2-acetic acid;
(S)-2,3,4,5-Tetrahydro-7-rt[(benz; ., .i~7.ol-2-yl)methyl]amino~carbonyl]-3-oxo-lH-1,4-berl7.orli:~7çpine-2-acetic acid;
(+)-5-[[2,3,4,5-Tetrahydro-7-~[[(benzimidazol-2-yl)methyl]amino]calbollyl]-3-oxo-4-(2-phenylethyl)- 1 H- 1 ,4-benzodiazepin-2-yl]methyl]tetra_ole;
(S)-2,3,4,5-Tetrahydro-7-[[[(4-aza-5-methylbel.~,i..licl~7.ol-2-yl)methyl]amino]carbonyl]-3-oxo- 1 H- 1 ,4-benzodiazepine-2-acetic acid;
(+)-2,3,4,5-Tetrahydro-7-[3-(ben7.imicl~7ol-2-yl)propyl]-4-methyl-3-oxo-lH-1,4-benzodiazepine-2-acetic acid;
(+)-2,3,4,5-Tetrahydro-7-[[N-[(benzill~dazol-2-yl)methyl]-N-(4-aminobutyl)amino]carbonyl] -3-oxo-4-(2-phenylethyl)- 1 H- 1 ,4-benzodiazepine-2-acetic acid;
(+)-2,3,4,5-Tetrahydro-7-[[[(ben7imi~ 7.ol-2-yl)methyl]amino]carbonyl]-4-methyl-3-oxo- lH- 1 ,4-benzodiazepine-2-(N-hydroxy)aret~mide;
Ethyl (+)-3-[[[2-(Ben7.imi~7.~1-2-yl)ethyl]amino]succinoyl]amino-4-pentynoate;
(+)-3-[[[2-(Bçn7.imi(1~7.ol-2-yl)ethyl~amino]succinoyl]amino4-pentynoic acid;
(+)-2,3 ,4,5-Tetrahydro-7-[[N-[(benzimidazol-2-yl)methyl] -N-[[4-(4-azido-3-iodo-2-hydroxybenzoyl)amino]butyl] amino]carbonyl] -3 -oxo-4-(2-phenylet'nyl)- 1 H-1,4-benzodiazepine-2-acetic acid;
2,3,4,5-Tetrahyciro-7-~[[(iS)-(ben7.imi~l~7.Qi-2-yi)ethyi]amino3carbonyl]-4-methyl-3-oxo- 1 H- 1 ,4-benzodiazepine-(2S)-acetic acid;
2,3,4,5-Tetrahydro-7-[[[( 1 R)-(benzimidazol-2-yl)ethyl]amino]carbonyl3-4-methyl-3-oxo-lH-1,4-benzodiazepine-(2S)-acetic acid; and .
w o 97n4119 PCTrUS96/20748 (+)-7-[[[(4,5-Dimethyl-lH-imi~1~7~1-2-yl)methyl]methyla~mino]carbonyl~-2,3,4,5-tetrahydro-4-methyl-3-oxo- lH- 1 ,4-benzodiazepine-2-acetic acid;
or pharmaceutically acceptable salts thereof.
The most preferred fibrinogen receptor antagonist template is of the sub-S formula (VIa), wherein CR2R2 is CHCH2CO2H, CR3R3 is C=O, and CRsRs isCH2. Vitronectin fibrinogen receptor antagonism is particularly pronounced when the A-W- substituent is attached to the 7-position of the 3-oxo-2,3,4,5-tetrahydro-lH- 1 ,4-benzodiazepine ring system.
In the formula below the definitions for the ~.ub~.LiLuents are as defined in 10 ft rm~ (I)-(V) and (XX)-(XXI), unless specified otherwise.
Another embodiment of a preferred fibrinogen receptor template A is represented by the 1,4-benzodiazepine 2,5-dione of sub-formula (VII);
~Y
R (VII) 1 5 wherein:
Y is H, Cl 4alkyl, Cl 4alkoxy, Cl 4alkoxycarbonyl, F, Cl, Br, I, CF3, oRf, S(O)kRf, CORf, NO2, N(Rf)2, CO(NRf)2, CH2N(Rf)2, methylenedioxy, CN, CO2Rf, OC(O)Rf, or NHC(O)Rf; and Rh is (CH2)qC02R
The plepal~tion and the use of tnis sub-structure in ~l~ing fibrinogen receptor antagonists of this sub-formula is detailed in Bondinell, et al., WO
93/OOO9S published January 7, 1993 and Blackburn, et al., WO 93/08174, publishedApril 29, 1993.
..
W O 97/24119 PCT~US96/20748 Table I, below, sl~mm~ries other ~l~felled fibrinogen receptor templates that are included within the scope of the present invention. Such templates are:
Table I
(VIII) Ais~R ~R22 J~R2~ j~R22 or / ~R
whGreill:
R21 and R22 independently are H or -Z-CO2Rfor Z-CON~Rf)2 with the proviso that one of Al or A2 is -ZCO2Rf or Z-CON(Rf)2;
Zis-CH2-,~O(CH2)q~~~NRf(CH2)q~~~S(CH2)q~-CH2CH2-,-CH(CH3)CH2-, -(CH2)3-, -CH=CH-, -C(CH3)=CH-, CH2-CH=CH- or CH=CHCH2; and Y is H, Cl 4alkyl, Cl 4alkoxy, Cl 4alkoxycarbonyl, F, Cl, Br, I, CF3, oRf, S(O)kRf, CORf, NO2, N(Rf)2, CO(NRf)2, CH2N(Rf)2, methylenedioxy or Z-CORf, in Alig, et al., EP 0 381 033, published August 8, 1990.
(IX) R6SO2NH CO2R~
20 wherein:
R6 is aryl, Cl lOalkyl, C3 6cycloalkyl, C4 1oaraLkyl, Cl loalkoxyalkyl, Cl loalkaryl, Cl loalkylthio~lkyl, Cl loaLkoxythioalkyl, Cl loalkylarnino, C4 1 oaralkylarnino, C 1-1 oalkanoylarnino, C4_ 1 oaralkanoylarnino, C 1-1 oaL~anoyl, C4 10aralkanoyl, or Cl locarboxyaLkyl; and S Y is H, Cl 4alkyl, Cl 4aLlcoxy, Cl 4alkoxycarbonyl, F, Cl, Br, I, CF3, ORf, S(o)kRf, CORf, NO2, N(Rf)2, Co(NRf)2, CH2N(Rf)2, methylenedioxy, CN, CO2Rf, OC(o)Rf, or NHC(O)Rf, in Egbertson, et al., EP 0 478 328, published April 1, 1992.
(X) --M~\M2~G~ CHCO2Rf wherein:
Ml is CH or N;
M2 is CH or N, with the proviso that when Ml is CH, M2 is N; and G' is N or N~3R", in Eldred, et al., EP 0542 363, published May 19, 1993.
. (XI) --M~\M2{><0H
--/ CH2CO2R~
wherein:
Ml is CH or N; and M2 is CH or N, with the proviso that when Ml is CH, M2 is N, in Porter, et al., EP 0 537 380, published April 21, 1993.
I
W O 97t24119 PCTrUS96/20748 ~XIIl ~3 . M ~ Rh wherein:
M1 isCHorN;
S Y is H, Cl 4alkyl, Cl 4alkoxy, Cl 4alkoxycarbonyl, F, Cl, Br, I, CF3, ORf, S(O)kRf, CORf, NO2, N(Rf)2, CO(NRf)2, CH2N(Rf)2, methylenedioxy, CN, Co2Rf, OC(O)Rf, or NHC(o)Rf;
D3 is CH2 or C=O; and Rh is (cH2)qco2Rf7 10in Klinnick, et al., EP 0 635,492, published January 25, l99S.
(XIII) \B~ N
wherein:
Y is H, C~ 4alkyl, C1 4alkoxy, Cl 4alkoxycarbonyl, F, Cl, Br, I, CF3, oRf, lS S(o)kRf, CORf, NO2, N(Rf)2, Co(NRf)2, CH2N(Rf)2, methylenedioxy, CN, CO2Rf, OC(O)Rf, or NHC(O)Rf;
Rh is ~CH2)nCO2Rf; and B is HsC)~ , ~N H3C ~
- in Blackburn, et al., WO 9S/04057, published February 9, 1995.
~V) W O 97/24119 PCT~US96/20748 ,~
N~ L -C02R9 O
wherein:
L* is -C(O)NRg-(CH2)-, ~C(O)~(CH2)q~~ NRg-(CH2)q-, ~O~(CH2)q~~ or S(O)k-(CH2)q~~
in Hartman, et al., EP 0 540 331, published May 5, 1993.
~V) N N--CH2 Co2R9 CO~o in Sugihara, et al., EP 0 529,858, published March 3, 1993.
(XVI~
~~
~ "'--CO R~
wherein:
Y is H, Cl 4alkyl, Cl 4alkoxy, Cl 4alkoxycarbonyl, F, Cl, Br, I, CF3, ORf, S(o)kRf, CoRf1, N02, N(Rf)2, Co(NRf)2, CH2N(Rf)2, methylenedioxy, CN, CO2Rf, Oc(o)Rf~ or NHC(O)Rf, in ~imm~i~b~h, et al., EP 0 483 667, published May 6, 1992.
(XVII) ~(CH2)oco2Ro N
in Linz, et al., LP 0 567 968, published November 3, 1993.
(XVIII) CA 0224l633 l998-06-26 W ~ 97/2~119 PCTAUS96/20748 Z"' Z~
~<Co2R9 wherein:
Rd is Het-C0 6alkyl; and Z", Z"' independently are hydrogen, Cl 4alkyl, halo, ORf, CN, S(o)kRf, 5 Co2Rf, or OH, in Bovy, et al., EP 0 539 343, published April 28, 1993.
Compounds of this invention comprising specific fibrinogen templates are named in the examples. These examples illustrate the invention, but do not limit the 10 scope of the invention.
The above descriptions of fibrinogen receptor templates for use in the present invention were taken from pending published patent applications. Reference should be made to such patent applications for their full disclosures, including the methods of plc~ali~lg said templzlt~s and specific compounds using said templates, the entire 15 disclosure of such patent applications being incol~oldl~d herein by reference.
Table II, below, describes other fibrinogen receptor antagonists, whose core structures would be useful in carrying out the instant invention. Reference should be made to the patent applications and other publications for their full disclosures, 20 including the methods of preparing said tt-mpl~t~s and specific compounds using said templates, the entire disclosure of the noted patent applications and otherpublications being incorporated herein by reference. Since it is contemplated that any fibrinogen receptor antagonist that is linked to an optionally fused nitrogen-cont~ining five-m.-.mhP.red ring will possess the novel utility described herein, the 25 list below does not limit the scope of the present invention.
WO 97124119 PCT~US96/20748 Table II
Adir et Col~r:-~nie FR 928004, June 30, 1992, Fauchere, J. L., et al.
EP 0578535, June 29, 1993, Fauchere, J-L, et al.: Describes X-RGDW-OH analogs, where X contains a cationic arnine.
CA 2128560, Jan. 24, 1995, Godfroid, J-J, et al., substituted pipel~ines.
Asahi Breweries, Ltd.
JP 05239030, Sep. 17, 1993, aminomethyltetrahydroisoquinolines.
Asahi Glass WO 90/02751, Ohba, M. et al.: Sept. 8, 1989: Describes cyclic RGD-con~ining peptides.
WO 90/115950, Mar. 22, 1990, Ohba, M., et al.
EP 0406428, 1/9/91: Describes cyclic RGD-cont:~ining peptides WO 92/09627, Isoai, A. et al.: Nov. 29, 1991: Describes cyclic RGD-cont~ining peptides.
~s~c~PIls AG
DE 4207254, (Der 93-289298/37) Mar. 7, 1992, Zoller, G., et al.: Describes guanidinopropyl-4-oxo-2-thioimida_olidin-3-yl-Asp-X analogs EP 93904010, Feb. 24, 1993, Zoller, G., 4-oxo-2-Thioxoimi~1~7Olidine Derivatives.
EP 0565896, Mar. 18, 1993, Klinger, O, et al.: Describes guanidinoethylphenyloxyacetyl-Asp-X analogs.
EP 0566919, (Der 93-338002/43) Apr. 3, 1993, Zoller, G., et al.: Describes guanidinopropyl-4-oxo-2-thioimidazolidin-3-yl-Asp-X analogs.
EP 580008, (Der 94-027663/04) July 6, 1993, Zoller, G., et al.: Describes S-m-guanidinophenyl-2,4-dioxoimi(l~7.Qlidin-3yl)acetyl-Asp-Phg.
DE 224414, July 6, 1993, Zoller, G., et al.: Describes 5-m-~tl:~ni~inophenyl-2,4-dioxoimidazolidin-3yl)acetyl-Asp-Phg.
W O 97/24119 PCT~US96/20748 EP 584694, (Der 94-067259/09) Apr. 2, 1994, Zoller, G., et al.: Describes 5-m-guanidinophenyl-2,4-dioxoimidazolidin-3yl~acetyl-Asp-Phg .
" DE 4301747, (Der 94-235891/29) Jul. 28, 1994, Zoller, G., et al.: Describes 5-m-guanidinophenyl-2,4-dioxoimi(1~7O1idin-3yl)acetyl-Asp-Phg analogs.
DE 4308034, (Der 94-286666/36) Sept. 15, 1994, Klinger, O. et al.: Describes 5-m-guanidinophenyl-2,4-dioxoimi~i~7O1idin-3yl)acetyl-Asp-Phg analogs.
DE 4309867, Sept. 29, 1994, Klingler, O, et al.: Describes 5-m-guanidinophenyl-2,4-dioxoimi~ 7.olidin-3yl)acetyl-Asp-Phg.
Chiron WO 93/07169, ~Der 93-134382/16), Mar. 15, 1993, Devlin, J. J., et al.: Describes RGD peptides.
Ciba Geigy EP 0452210, (Der 91-305246/42) Apr, 5, 199Q, describes aminoalkanoyl-GDF
analogs.
EP 0452257, Mar. 26, 1991, Allen, M. C., et al.: Describes aminoalkanoylAsp-Phe analogs.
COR Ther~renti~c WO 90/15620, June 15, 1990: Describes cyclic RGD-cont~ining peptides.
EP 0477295, Apr. 1, 1992: Scarborough, R. M. et al.
WO 92/08472, May 29, 1992, Scarborough, R. M. et al.
WO 93/223356, April 27. 1993, Swift, R. L., et al.: Describes cyclic RGD-c--nt~inin~ peptides.
EP 0557442, Sept. 1, 1993, Scarborough, R. M., et al.
Scarborough, R. M.; Rose, J. W.; Hsu, M. A.; Phillips, D. R.; Fried, V. A.;
Campbell, A. M.; N--nni77i, L.; Charo, I. F., Barbourin, A GPIIb-IIIa-Specific Integrin Antagonist from the Venom of Sistrurus M. Barbouri, J.
BioL Chem., 266, 9359, 1991.
CA 0224l633 l998-06-26 W O97/24119 PCT~US96/20748 Daiichi Pharm Co Ltd.
JP 05078344-A, ~Der 93-140339/17) Mar. 30, 1993: Describes Bis-amidinoheter~cycles, eg. benzofurans.
S DuPont Merck WO 93/07170, Apr. lS, 1993: Describes cyclic-RGD-con~ining peptides.
WO 94/11398, May 26, 1994: Wells, G. J. et al. Describes cyclic RGD cont~ining peptides.
IL 109237, Ju1. 31, 1994.
WO 94/22909, (Der 94-333113/41) Oct. 13, 1994: DeGrado W. F., et al.
WO 94/22910, (Der 94-333114/41 Oct. 13, 1994: DeGrado W. F., et al. Prodrugs.
4, (Der 94-332838/41) Oct. 13, 1994: DeGrado W. F., et al. Cyclic peptides EP 625164, Nov. 23, 1994: Degrado, W. F., et al. Cyclic peptides.
Mousa, S. A.; Bozarth, J. M.; Forsythe, M. S.; Jackson, S. M.; Leamy, A.; Diemer, M. M.; Kapil, R. P.; Knabb, R. M.; Mayo, M. C.; Pierce, S. K.; al., e., ~ntiplzltelet and An-ilhloll-botic Efficacy of DMP 728, a Novel Platelet GPIIb/IIIa Receptor Antagonist, Circulation, 89, 3, 1994.
~ackson, S.; DeGrado, W.; Dwivedi, A.; Parthasarathy, A.; Higley, A.; Krywko, J.;
Rockwell, A.; Markwalder, J.; Wells, G.; Wexler, R.; Mousa, S.; Harlow, R., Template-Constrained Cyclic Peptides: Design of High-Affinity T .ig~n~1~ for GPIIb/IIIa, J. Amer. Chem. Soc., 116, 3220, 1994 Ellem Ind Farma Spa GB 2207922, Aug, 3, 1988, describes linear RGD analogs.
Farmitalia Erba SRL Carlo EP 611765 (Der 94-265375/33), Aug 24, 1994: Cozzi, P., et al. Describes 5-(2-~y~ lylmethyl-2-imi~ 7Ql-l-yl)-1-cyclohexylethylidene)aminoxypentanoic acid.
W O 97124119 PCTrUS96/20748 Fuji Photo Film JP 04208296-A (Der. 92-303598/38), Nov. 30, 1990, Describes RGD peptides.
JP 04213311-A (Der. 92-305482/38), Nov. 27, 1990, Describes mllltimeric RGD
peptides.
JP 04217693-A, (Der 92-312284/38), Oct. 23, 1990, Descirbes mllltim~ric RGD
peptides.
JP 04221394-A (Der. 92-313678/38), Oct. 26, 1990, Describes mllltimf~ric RGD
peptides.
JP 04221395-A (Der. 92-313679/38), Oct. 26, 1990, Describes m-lltim~ric RGD
peptides.
JP 04221396-A (Der. 92-313680/38), Oct. 26, 1990, Describes mllltim~ric RGD
peptides.
JP 04221397-A (Der. 92-313681/38), Dec. 20, 1990, Describes mllltim~ril~ RGD
peptides.
EP 0482649 A2, April 29, 1992, Kojima, M. et al.: Describes RGD peptides.
EP 0488258A2, June 3, 1992, Komazawa, H., et al: Describes RGD peptides.
EP 503301-A2, Feb. 14, 1991, Kitaguchi, H. et al.: Describes RGD peptides.
JP 05222092, May 21, 1993, Nishikawa, N., et al.: DescribesT in~r X-RGDS.
JP 06239885, (Der 94-313705/39), Aug 30, 1993, Nishikawa, N. et al.: Describes mllltim~ric RGD peptides.
WO 9324448, (Der 93-405663/50), Dec. 9, 1993, Nishikawa, N., et al.: Describes ml-ltimf~ric retro-inverseo RGD peptides.
JP 06228189, (Der 94-299801/37), Aug. 16, 1994. Describes RGD peptides.
EP 619118, (Der 94-311647/39), Oct. 12, 1994, Nishikawa, N. et al.: Describes linear RGD peptides.
Fujisawa EP 0513675, May 8, 1992, N. Umekita, et al.: Describes amidinophenyloxyalkanoyl-Asp-Val-OH analogs.
WO 9409030-A1, Apr. 28, 1994, T~k~ gi, H., et al.: Describes Amidinophenoycbutanoyl-Asp-Val-OH analogs.
W O 97/24119 PCT~US96/20748 EP 0513675, (Der 92-383589/47): Describes Ami~lin~phenyloxybutyrl-Asp-Val analogs.
WO 9500502, Jan, 5, 1995, Oku, T., et al.,: Describes "aminopi~e~ e derivatives."
FR 144633: Thromb Haem. 69, 706, 1993.
Cox, D.; Aoki, T.; Seki, J.; Motoyama, Y.; Yoshida, K., Pent~mitline: A Speci~lcNonpeptide GPIIb/IIIa Antagonist, Thromb. Haem., 69, 707, 1993.
G~nPntef h WO 90/15072 (Der 91007159): Describes RGD-cont~inin~ peptides:
WO 91/01331 (Der91058116), July 5, 1990, P. L. Barker, et al.: Describes cyclic RGD-corltziinin~ peptides WO 91/04247, Sept. 24, 1990, T. R. Webb: Describes (guanidinoalkyl)Pro-GD
analogs.
WO 91/11458 (Der 91252610), Jan. 28, 1991, P. L. Barker, et al.: Describes cyclic RGD-cont~inin~ peptides WO 92/07870, Oct. 24, 1991 J. P. Burnier, et al.: Describes cyclic RGD-c-n~ining peptides.
WO 92/17492, Oct. 15, 1992, Burnier, J. P. et al.: Describes cyclic RGD-cont~ining peptides.
CA 2106314, Oct. 6, 1992, Burnier, J. P. et al.
WO 93/08174, Oct. 15, 1991, B. K. Blackburn, et al.: Describes 2,5-dioxo-1,4-ben70fliz~7~pines.
CA 2106314, Oct. 6, 1992, Burnier, J. P., et al.
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WO 91/15515, (Der 91-325173/44) Apr. 6, 1990, describes eyelic RGD analogs.
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AU A-86926/91, May 7, 1992, Hirnmelsbach, F. et al.: Describes amidinophenyl compounds.
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DE 4124942, Jan. 28, 1993, Hirnmelsbach, F. et al.: Describes Ami~lin---triarylproionic acid analogs.
DE 4129603, Mar. 11, 1993, Pieper, H, et Al.: Describes amidinobiphenyl-I~ç ~iT.~itlA7ole.
W O 97/24119 PCT~US96/2~748 EP 0547517 A1, (Der 93-198544) ~une 23, 1993, Soyka, R., et al.: Describes pyridyl compounds.
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W O 97124119 PCTrUS96/20748 Fuji Photo Film JP 04208296-A (Der. 92-303598/38), Nov. 30, 1990, Describes RGD peptides.
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EP 503301-A2, Feb. 14, 1991, Kitaguchi, H. et al.: Describes RGD peptides.
JP 05222092, May 21, 1993, Nishikawa, N., et al.: DescribesT in~r X-RGDS.
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WO 9500502, Jan, 5, 1995, Oku, T., et al.,: Describes "aminopi~e~ e derivatives."
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WO 91/11458 (Der 91252610), Jan. 28, 1991, P. L. Barker, et al.: Describes cyclic RGD-cont~inin~ peptides WO 92/07870, Oct. 24, 1991 J. P. Burnier, et al.: Describes cyclic RGD-c-n~ining peptides.
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W O 97/24119 PCTrUS96/207~8 Dennis, M. S.; Henzel, W. J.; Pitti, R. M.; T., L. M.; Napier, M. A.; Deisher, T. A.;
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JP 05078244-A, Mar. 30, 1993: Describes dibenzo(b,e)oxepine derivatives.
CA 0224l633 l998-06-26 W O97/24119 PCTrUS96/20748 Laboratoire Chauvin WO 9401456, Jan. 20, 1994, Regnouf, D. V. J. et al.: Describes Ac-Arg-Gly-Asp-NHBn analogs.
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Merck EP 0368486 (Der 90-14g427/20), Nov. 10, 1988: Describes X-R-Tyr-D-Y analogs.
EP 0382451 (Der 90248531): Descirbes RGD-cont~ining snake venom inhibitors.
EP 03~538 (Der 90248420): Descirbes RGD-cont~ining snake venom inhibitors.
EP 0410537, July 23, 1990, R. F. Nutt, et al.: Describes cyclic RGD-cont~ining peptides.
EP 0410539, July 25, 1990, R. F. Nutt, et al.: Describes cyclic RGD-cont~inin~
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EP 0410540, July 25, 1990, R. F. Nutt, et al.: Describes cyclic RGD-cont~inin~
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EP 0410541, July 25, 1990, R. F. Nutt, et al.: Describes cyclic RGD-cont~inin~
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EP 0410767, July 26, 1990, R. F. Nutt, et al.: Describes linear RGD-cont~ining peptides.
W O 97/24119 PCT~US96/20748 EP 0411833, July 26, l99Q R. F. Nutt, et al.: Describes cyclic RGD-con~ining peptides.
EP 0422937, Oct. 11, 1990, R. F. Nutt, et al.: Describes cyclic RGD-cont~ining peptides.
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EP 0487238, Octover 13, 1991, T. M. Connolly, et al.: Describes Linear RGD-cont~ining EP 0437367 (Der 91209968), M. Sato et al.: Describes cyclic RGD-con~ i peptides, as inhibitors of osteoclast-m~ tl-A bone resorption.
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WO 9409029, Apr. 28, 1994, Nutt, R. F. and Veber, D. F., describes piperidinylethylpyrrolidinylacetyl-Asp-Trp(tetrazoles) .
EP 618225, (Der 94-304404/38) Oct. 5, 1994, Describes RGD peptide analogs as antimetastatic colllpou~ds.
DE 4310643, (Der 94-311172/39), Oct. 6, 1994, Jonczyk, A.. et al.,: Describes cyclic RGD analogs as :~ntimt-,t~t~ti~, agents.
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EP 0479481, Sept. 25, 1991, M. E. Duggan et al.: Describes X-GlyAsp-Y linear semipeptides.
EP 0478328, Sept. 26, 1991, M. S. Egbertson, et al.: Describes tyrosine derivatives.
EP 0478362, Sept. 27, 1991 M.3~. Duggan et al.: Describes X-Gly-(3-phenethyl),BAla analogs.
EP 0478363, Sept. 27, 1991, W. L. Laswell, et al.: Describes Tyrosine sulfon~mil:1ec .
EP 0512829, May, 7, 1992, Duggan, M. F., et al.: Describes chiral 3-hydroxy-6-(4-piperidinyl)heptanoyl-,B-X-,B-Ala-OH analogs, with variations on X and the central alkanoyl chain.
W O 97/24119 PCT~US96/20748 EP 0512831, May, 7, 1992, Duggan, M. E., et al.: Describes chiral 2-oxo-3-(Piperidinylethyl)piperidinylacetyl-~-X-13-Ala-OH analogs, with variations on X and the central piperidinyl ring.
EP 0528586, August 5, 1992, M. S. Egbertson, et al.: Describes tyrosine sulfon~mi(1~s as inhibitors of osteoclast-mediated bone resorption.
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EP 0540334, October 29, 1992, G. D. Hartman, et al.: Describes bç~ oles.
US 5227490, Feb. 21, 1992, G. D. Hartman, et al.: Describes Tyrosine sulfonamides.
CA 2088518, Feb. 10, 1993, Egbertson, M. S., et la. aminoalkyl-phenyl derivs. as bone resorption inhibts.
US 5206373-A, (Der 93-151790tl8) Apr. 27, 1993, Chung, J. Y. L., et al.: Describes MK-383-type compounds.
WO 9316994, (Der 93-288324/36), Sep. 2, 1993, Chung, J. Y. L., et al.: Describes pyridinylbutyl-L-Tyrbutylsulfon~mi~le US 5264420-A, Nov. 23, 1993, Describes piperidinylalkyl-Gly-betaAla analogs.
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GB 945317 A, Mar. 17, 1994 (Priority US 34042A, Mar. 22, 1993).
GB 2271567 A, Apr. 20, 1994, Hartman, G. D. et al.: Describes compounds replacing Tyr with beta-phenylsuccinate.
US 5294616, (Der 94-091561/11) Mar. 15, 1994, Egbertson, M. S., et al.
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US 5312923, May 17, 1994, Chung, J. Y. L. et al.
W O97/24119 PCT~US96/20748 HU 9400249, May 30, 1994, Gante, J. et al.,: Describes pi~c-~ine analogs.
WO 9412181, (Der 94-199942/24), Jun. 9, 1994, Egbertson, M. S. et al.,: Describes piperidinylethyloxyphenyl acetic acid analogs , US 5321034, June 14, 1994, Duggan, M. E., et al.: Describes Piperidinylalkyl-bet~mino acids.
US 5334596, Aug. 2, 1994, Hartman, G. D. et al.
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GB 2276384, (Der 94-287743/36) Sep. 28, 1994, Claremon, D. A.., Liverton, N..,:
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Nippon Steel Corp WO 9405696, Mar. 17, 1993, Sato, Y., et al,.
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ONO Phar JP 05286922 (Der 93-383035/48), Describes guanidinophenol alkylbenzoic acid esters.
Roche EP 038,362, Feb. 19, 1990, M. Muller, et al.: Describes X-NHCHYCO-Gly-Asp-NHCHZCO2H analogs.
EP 0372486, June, 13, 1990, Allig, L., et al.
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US 5273982-A, (Der94-006713/01) Dec. 28, 1993: Describes arnidinophenyl-linked S Gly-Asp-X se~ ~Lides.
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WO 91/04746, S. I. Klein et al.: Describes des-ArninoArginine RGD analogs.
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WO 93/11759, Dec. 7, 1992, S. I. Klein et aL: Describes Bis-gn~nitlinoaklanoic acid analogs.
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CA 2107088, Sept. 29, 1992, Klein, S.I. et al.
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~n cases wherein the compounds of this invention may have one or more chiral centers, unless specified, this invention includes each unique nonracemiccompound which may be synthesized and resolved by conventional techniques. In cases in which compounds have unsaturated carbon-carbon double bonds, both the cis (Z) and trans (E) isomers are within the scope of this invention. The m~z~nin~ of 15 any substituent at any one occurrence is independent of its m~ning, or any other substituent's m~ning, at any other occurrence.
Abbreviations and symbols commonly used in the peptide and chemical arts are used herein to describe the compounds of this invention. In general, the amino acid abbreviations follow the IUPAC-IUB Joint Commission on Biochemical 20 Nomenclature as described in Eur. J. Biochem., 158, 9 (1984).
Cl 4alkyl as applied herein means an optionally substituted alkyl group of 1 to 4 carbon atoms, and inc.lucles methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl and t-butyl. Cl 6alkyl additionally includes pentyl, n-pentyl, isopentyl, neopentyl and hexyl and the simple aliphatic isomers thereof. C0 4alkyl and C0 6alkyl 25 additionally indicates that no alkyl group need be present ~e.g., that a covalent bond is present).
Any Cl 4alkyl or Cl 6 alkyl, C2 6 alkenyl, C2 6 alkynyl or Cl 6 oxoalkyl may be optionally substituted with the group RX, which may be on any carbon atom that results in a stable structure and is available by conventional synthetic techniques.
30 Suitable groups for Rx are Cl 4alkyl, OR, SR, Cl 4alkyl, Cl 4alkylsulfonyl, W O 97/24119 PCTrUS96/20748 C I 4alkylsulfoxyl, -CN, N(R )2, CH2N(R )2. -N02, -CF3, -C02R -CON(R )2~ -COR, -NR C(O)R, OH, F, Cl, Br, I, N3, or CF3S(O)~-,wherein r is O to 2.
Ar, or aryl, as applied herein, means phenyl or naphthyl, or phenyl or naphthyl substituted by one to three subctitllent~ sueh as those defined above for 5 alkyl, espeeially Cl~alkyl, Cl 4alkoxy, Cl 4alkthio, trifluoroalkyl, N3, OH, C02H
F, Cl, Br or I.
Het, or heterocyele, in~lic~tl~.s an optionally ~llb~liLuled five or six membered monoeyelic ring, or a nine or ten-membered bieyelie ring cf-nt~ining one to three heteroatoms ehosen from the group of nitrogen, oxygen and sulfur, whieh are stable 10 and available by eonventional ehPmic~l synthesis. Illustrative heterocycles are benzofuran, ben7imicl~01e, bellzoL)yl~l, benzothiophene, biotin, furan, imicl~ole, indoline, morpholine, piperidine, ~iL~t;ld~ille, pyrrole, pyrro}idine, pyridine,tetrahydropyridine, pyridine, thiazole, thiophene, quinoline, isoquinoline, and tetra-and ~lhyd~ quinoline and isoquinoline. Any ~cçc.cihle combination of up to 15 three substit~entc on the Het ring, such as those defined above for alkyl that are available by ehemieal synthesis and are stable are within the seope of this invention.
C3 7cyeloaL~yl refers to an optionally substituted carbocyelic system of three to seven earbon atoms, whieh may eontain up to two unsaturated earbon-earbon bonds. Typieal of C3 7eyeloalkyl are eyelopropyl, cyclobutyl, cyclopentyl, 20 cyelopentenyl, eyelohexyl, eyelohexenyl and eyeloheptyl. Any eombination of up to three substi~uent.~, such as those defined above for alkyl, on the cycloalkyl ring that is available by conventional chemieal synthesis and is stable, is within the seope of this invention.
When Rb and Rc are joined together to form a five- or six-membered 25 aromatie or non-aromatic carbocyclic or heterocyclie ring fused to the ring to whieh Rb and Rc are attached, the ring formed will generally be a five- or six-membered heterocycle s~lecte~ from those listed above for Het, or will be a phenyl, eyclohexyl or cyclopentyl ring. Preferably Rb and Rc will be -D1=D2-D3=D4 wherein Dl - D4 are independently CH, N or C-RX with the proviso that no more than two of D 1 - D4 W O 97/24119 PCT~US96/20748 are N. Most preferably, when Rb and Rc are joined together they form the group -CH=CH-CH=CH-.
- Certain radical groups are abbreviated herein. t-Bu refers to the tertiary butyl radical, Boc refers to the t-butyloxycarbonyl radical, Fmoc refers to the 5 fluorenylmethoxycarbonyl radical, Ph refers to the phenyl radical, Cbz refers to the - benzyloxycarbonyl radical, BrZ refers to the o-bromobenzyloxycarbonyl radical, ClZ refers to the o-chlorobenzyloxycarbonyl radical, Bzl refers to the benzyl radical, 4-MBzl refers to the 4-methyl benzyl radical, Me refers to methyl, Et refers to ethyl, Ac refers to acetyl, Alk refers to Cl 4alkyl, Nph refers to 1- or 2-naphthyl and cHex 10 refers to cyclohexyl. Tet refers to 5-tetrazolyl.
Certain reagents are abbreviated herein. DCC refers to dicyclohexylcarbo-liimi(~, DMAP refers to dimethylaminopyridine, DIEA refers to diisopropylethyl amine, EDC refers to 1-(3-dimethylaminopropyl)-3-ethylcarborliimi(le, hydrochloride. HOBt refers to l-hydroxybenzotri~7Ole, THF
15 refers to tetrahydrofuran, DIEA refers to diisopropylethylamine, DME refers to dimethoxyethane, DMF refers to dimethylform~micle, NBS refers to N-bromosuccinimide, Pd/C refers to a palladium on carbon catalyst, PPA refers to 1-propanephosphonic acid cyclic anhydride, DPPA refers to diphenylphosphoryl azide, BOP refers to benzotriazol-l-yloxy-tris(dimethyl-amino)phosphonium 20 hexafluorophosphate, HF refers to hydrofluoric acid, TEA refers to triethylamine, TFA refers to trifluoroacetic acid, PCC refers to pyridinium chlorochromate.
Compounds of the formula ~ (V) are prepared, for example, by reacting a compound of formula (XIX) with a compound of formula (XX), wherein Ll and L2 are groups which may react to fo;m a covalent bond in the moiety W, by methods 25 generally known in the art.
~ ~> L~ > W-A
RC N + Ll-A RC N
(XIX) (XX) (I) Typical methods include coupling to form amide bonds, nucleophilic displ~rçm~nt reactions and p~ m catalyzed couplings. For instance, when W
W O 97/24119 PCTrUS96/20748 contains an ether or amine linkage, the bond may be formed by a displacement reaction, and one of Ll and L2 will contain an amino or hydroxy group and the other will contain a displaceable group, such as a chloro, bromo or iodo group. When Wcontains an amide bond, typically one of Ll and L2 will contain an amino group, and 5 the other will contain a carboxylic acid group. In another approach, Ll may be an aryl or heteroaryl bromide, iodide or trifluoromethylsulfonyloxy derivative and L2 may contain an amino group and the amide linkage may be formed by p~ m-catalyzed aminocarbonylation with carbon monoxide in a suitable solvent such as dimethylform~micle or toluene.
It will be apparent that the precise identity of Ll and L2 will be dependent upon the site of the linkage being }'ormed. General methods for preparing the linkage -(CHR")~-U-(CHR")s-V- are described, for example, in EP-A 0 372 486 and EP-A 0 381 033 and LP-A 0 478 363, which are incol~oldt~d herein by reference.
For in~t~nre, if V is CONH, Ll may be -NH2, L2 may be O~I (as in an acid) 15 or Cl (as in an acid chloride), and R6 may be W-(CRr2)q-Z-(CR'Rl0)r~U-(CRI2) C(O), with any functional groups optionally protected. For example, R6 may be (benzyloxycarbonyl-amidino)benzoyl- or (N~Boc,NgUan-Tos)arginyl-. When L2 is OH, a coupling agent is used.
Similarly, if V is NHCO, Ll may be -CO2H or CO-Cl, L2 may be -NH2, and R6 may be W~(CR12)q~Z(CRIRl~)r-u-(cRl2)s-. For example, R6 may be (benzyloxycarbonyl-amidino)phenyl, (benzyloxycarbonylamino)methylbenzyl- or 6-(benzyloxycarbonylamino)hexyl- .
Where V is NHSO2, Ll may be SO2Cl, L2 may be -NH2 and R6 may be as above. Where V is SO2NH, Ll may be -NH2 and L2 may be SO2CI. Methods to 25 prepare such sulfonyl chlorides are disclosed, for in~t~n~e, in J. Org. ~he~l., 23, 1257 (1958).
If V is CH=CH, Ll may be -CHO, L2 may be CH=P-Ph3 and R6 may be W-(CR12)q-Z-(CRlRlO)~U-(CRl2)s-- ~l~rnzlt~ly, Ll may be CH=P-Ph3, L2 may be CHO, e.g., R6 may be W~(CR~2)q~Z~(CRlRl0)r-u-(cRl2)s-l-cHo.
-~6 -~ .
W O97/24119 PCTrUS96/20748 Where V is CH2CH2 may be obtained by reduction of a suitably protected compound wherein V is CH=C~I.
Where V is CH20, CH2N or C--C, L1 may be -OH, -NH or -C~ C H,respectively; L2 may be -Br; and R6 may be W~(CR~2)q~Z(CR~RI~)r~u-(cR~2)s-~ For example, R6 may be (benzyloxycarbonylarnino)-methylbenzyl- or 2-(N-benzyl-4-piperidinyl)-ethyl.
Similarly where U or V is OCH2, NR'CH2 or C~ C, Ll may be -CH2Br and L2 may be -OH, -NH or -C= C H, le~.~e.;lively. Alternately, when U or V is C--C, Ll maybe Br, I or CF3SO3, L2 may be C--C H and the coupling may be catalyzed by p~ m and a base.
Compounds wherein V is CHOHCH2 may be prepared from a suitably protected compound where V is CH=CH by the procedure disclosed in J. Org.
Chem., 54, 1354 (1989).
Compounds wherein V is CH2CHOH may be obtained from a suitably protected compound where V is CH=CH by hydroboration and basic oxidation as disclosed in Tet. Lett., 31, 231 (1990).
The core 6-7 fused ring system is prepared of formula (VI) by methods well known in the art, e.g., Hynes, et al., J. Het. Chem., 1988, 25, 1173; Muller, et al., Helv. Chim. Acta., 1982, 65, 2118; Mori, et al., Heterocycles, 1981, 16, 1491.
Similarly, methods for preparing benzazepines, 1,4-benzofhi~7.e.pin~ s, 1,4-benzoxazepines and 1,4-benzodiazepines are known and are disclosed, for in.~t~n~e, in Bondinell, et al., International Patent Application WO 93/00095.
The sl~htom~s which follow detail the ~l~dLion of the compounds of the instant invention.
W O 97/24119 PCT~US96/20748 S. ' - ~ I
CH30~ ~ CH~0~[~
o CO2Et CH30~ H0 ~
3 C02Et 4 C02Et TfO ~ HO C~
C02Et C02Et g_--~ N r ~ ~ h a) EtOAc/LiN(TMS)2, THF; b) Et3SiH, BF3 ~ OEtz, CH2Cl2; c) H2~ 10% Pd/C, EtOH;
d) EtSH, AICl3, CH2Cl2; e) Tf20, 2,6-h-ti~lin~., CH~Cl2; f) CO, KOAc, Pd(OAc)2, 5 dppf, DMSO; g) 2-(methylaminomethyl)l~e~ ole dihydrochloride, EDC, EIOBt ~ H20, (i-Pr)2NEt, CH~CN; h) 1.0 N NaOH, EtOH.
CA 0224l633 l998-06-26 An ~p,op.iately substituted deoxybenzoin, such as 2-(4-methoxyphenyl)-l-phenylethanone (Chem. Ber. 1958, 91, 755-759), is reacted in an aldol-type reaction with the enolate of ethyl acetate, which can be generated from ethyl acetate on exposure to an a~pr~p.iate amide base, for in~t~nre lithium diisopropylamide (LDA) 5 or lithium bis(trimethylsilyl)amide (LiN(TMS)2), to afford I-2. Fre~uently, THF is - the solvent of choice for an aldol reaction, although THF in the presence of various additives, for in.~t~nre HMPA or TMEDA, is often used. Reaction of I-2 with triethylsilane (Et3SiH) in the presence of boron trifluoride etherate (BF3 ~ OEt2) according to the general protocol of Orphanopoulos and Smonu (Synth. Commun.
10 1988, 833) for the reduction of tertiary benzylic alcohols affords I-3, together with the olefinic product derived from ~ limin:~tion of the alcohol. The olefinic product can be conveniently converted to I-3 by hydrogenation over a F~ m catalyst, such as palladium metal on activated carbon (Pd/C), in an ~pl~pliate inert solvent, for in~t:~nre mr.th~nol, ethanol, or ethyl acetate. Removal of the methyl ether of I-3 15 to give I-4 can be accomplished by reaction with eth~nethiol (EtSH) in the presence of a Lewis acid catalyst, preferably anhydrous ~lnminllm trichloride (AlCl3), in an inert solvent, for in~t~nce CH2Cl2. Other useful methods for removal of a methylether are described in Greene, "Protective Groups in Organic Synthesis" (published by Wiley-Interscience). Alcohol I-4 is converted to its trifluoromethanesulfonate 20 ester I-5 by reaction with trifluoromethanesulfonic anhydride (Tf2O) in the presence of a suitable non-nucleophilic amine base, such as 2,6-lutidine, in an inert solvent, generally CH2Cl2. I-~ reacts with carbon monoxide (CO) in the presence of potassium acetate, 1, l '-bis(diphenylphosphino)ferrocene (dppf), and a F~ lillmcatalyst, for in~t~nce palladium acetate (Pd(OAc)2), in a suitable solvent, preferably 25 DMSO, according to the general method described by Cacchi and Lupi (Tet. Lett.
1992, 33, 3939) for the carboxylation of aryl trifluorometh~nt-sulfonates. The resulting benzoic acid derivative I-6 is converted to an activated form of the carboxylic acid using, for example, EDC and HOBt, or SOCl2, and the activated form is subsequently reacted with an ~pl~,p.iate amine, for in~t~nre 2-30 (methylaminomethyl)methylben7.imi-l~7.ole dihydrochlnri~le, in a suitable solvent W O 97/24119 PCT~US96/20748 such as DMF, CH~Cl2, or CH3CN, to afford I-7. Df~pçn-lin~ on whether acid neutralization is required, an added base, such as diisopropylethylamine ((i-Pr)2NEt) or pyridine, may be used. Many additional methods for converting a carboxylic acid to an amide are known, and can be found in standard reference books, such as 5 "Compendium of Organic Synthetic Methods", Vol. I - VI (published by Wiley-Interscience), or Bodansky, "The Practice of Peptide Synthesis" (published by Springer-Verlag). The ethyl ester of I-7 is hydrolyzed using aqueous base, for example, LiOH in aqueous THF or NaOH in aqueous methanol or ethanol, and the intermediate carboxylate salt is acidified with a suitable acid, for instance TFA or 10 HCl, to afford the carboxylic acid I-8. ~lt~rn~tively, the interm~ t.o carboxylate salt can be isolated, if desired, or a carboxylate salt of the free carboxylic acid can be prepared by methods well-known to those of skill in the art.
W O 97/24119 PCT~US96/20748 Scheme II
~ HO2C~ t-BUO2C~0~ t-Bu02C~C02CH3 ~- 1 23 t-BuO2C ~ t-BuO2C ~ e t-Bu02C ~ t-Bu02C~,CH3) CON(CH3)2 CO2Et H~2C~ CON(CH3)2 ~ N~N~ "CH3)2 ~CO2Et (~NH CH3 CO2Et N~ N J~ CH3)2 ~ NH CH3 CO2H
a) isobutylene, TfOH, CH2C12; b) methyl acrylate, Pd(OAc)2, P(tol)3, (i-Pr)2NBt,propionitrile; c) H2, 10% Pd/C, MeOH, EtOAc; d) 1.0 N LiOH, THF, H2O; e) dimethylamine hydrochloride, EDC, HOBt ~ H20, (i-Pr)2NEt, CH3CN; f) LiN(TMS)2, - THF, then BrCH2CO2Et; g) TFA, CH2Cl2; h) 2-(methylaminomethyl)ben7.imi(1~7.ole dihydrochloride, EDC, HOBt H20, (i-Pr),NEit, CH3CN; i) 1.0 N LiOH, THF, H20.
W O 97/24119 PCTrUS96t20748 Commercially available 4-bromobenzoic acid (II-l) is converted to the tert-butyl ester II-2 by reaction with isobutylene in the presence of a catalytic arnount of an acid, such as trifluorom~thAn.osulfonic acid (TfOH) or sulfuric acid, in _n inert solvent, generally CH2Cl2 or diethyl ether. Alternative methods for the formation of 5 tert-butyl esters are described in Greene, "Protective Groups in Organic Synthesis"
(published by Wiley-Interscience). Other esters can be employed, as long as theyare compatible with the subsequent chemistry and can be removed selectively whendesired. A Heck-type reaction between II-2 and methyl acrylate affords II-3. Thegeneral conditions for the Heck reaction have been reviewed by Heck (Org.
lO Reactions 1982, 27, 345). For ~I-2, reaction with methyl acrylate in the presence of palladium ~II) acetate (Pd(OAc)2) and tri-ortho-tolylphosphine (P(tol)3) in an inert solvent, such as CH3CN, propionitrile, or toluene, in the presence of an ~p.o~,iate acid scavenger, such as diisopropylethyla~lnine ((i-Pr)2NEt), affords II-3. Reduction of the a,l3-unsaturated ester of II-3 to afford the saturated compound II-4 occurs 15 under standard hydrogenation conditions, for instance reaction with hydrogen gas in the presence of a suitable catalyst, preferably palladium metal on activated carbon (Pd/C), in an inert solvent, generally m(~thAn~l, ethanol, ethyl acetate, or mixtures thereof. The methyl ester of II-4 is hydrolyzed using aqueous base, for example,LiOH in aqueous THF or NaOH in aqueous methanol or eth_nol, and the 20 interm~-liAte carboxylate salt is acidified with a suitable acid, for in~t~n~e T~A or HCl, to afford the carboxylic acid II-5. Tnis is converted to an activated form of the carboxylic acid using, for example, EDC and HOBt, or SOC12, and the activated form is subsequently reacted with an a~3p~ iate a~nine, for in~tAnce dimethylamine hydrochloride, in a suitable solvent such as DMF, CH2CI~, or CH3CN, to afford II-6.
25 Depending on whether acid neutralization is required, an added base, such as diisopropylethylamine ((i-Pr)2NEt) or pyridine, may be used. Many additional methods for converting a carboxylic acid to an amide are known, and can be foundin standard reference books, such as "Compendium of Organic Synthetic Methods", Vol. I - VI (published by Wiley-Interscience), or Bodansky, "The Practice of Peptide 30 Synthesis" (published by Springer-Verlag). Reaction of II-6 with an amide base, for CA 0224l633 l998-06-26 W O 97/24119 PCT~US96/20748 in.~t:~nCe~ lithium bis(trimethylsilyl)amide (3_iN(TMS)2), sodium bis(trimethylsilyl)amide (NaN(TMS)2), pot~c~ m bis(trimethylsilyl)amide , (KN(TMS)2), or lithium diisopropylamide (LDA), in an inert solvent, generally THF
or ethylene glycol dimethyl ether (DME), affords an interm~ te amide enolate.
5 This is generally not isolated, but rather is reacted in situ with an electrophile, for example ethyl bromoacetate, to afford the alkylated product II-7. Various additives known to those of skill in the art, for in~t~n~e HMPA, teLlalllelhylethylen~ mine (TMEDA), or 1,3-dimethyl-3,4,5,6-tetrahydro-2(1H)-pyrimidinone (DMPU), can be used to improve the eff1ciency of the alkylation reaction. ). The tert-butyl ester 10 group of II-7 is removed under acidic conditions, generally with TFA or HCl, in an inert solvent, usually CH2Cl2, 1~4-dioxane, or l~ Lules thereof, to afford the acid II-8. Other useful methods for the removal of tert-butyl esters are described in Greene, "Protective Groups in Organic Synthesis" (published by Wiley-Interscience). II-8 is converted to II-9 under the general conditions described in Scheme I for the conversion of I-6 to I-7, and II-9 is converted to II-10 by the general conditions described in Scheme I for the conversion of I-7 to I-8.
Schem~. m ~_NH a~_NH O COzCH3 ~N CO H
~ NH N ~ ~ COzEt N ~ N ~ ~ H CO2H
g_NH O
a) 3-carbomethoxypropionyl chloride, (i-Pr)2NEt, CH2Cl2; b) 1.0 N NaOH, MeOH;
c) ethyl 3-amino-4-pentynoate, EDC, HOBt ~ H20, (i-Pr)2NEt, CH~CN, DMF; d) 1.0 N LiOH, THF, H20, CH3CN.
Readily available 2-(2-aminoethyl)be~,~.;.l.itl~7.ole is reacted with 3-1() carbomethoxy~lopionyl chloride in the presence of an al,~r.,l,liate acid scavenger, such as triethylamine, diisopropylethylamine, or pyridine, in a neutral solvent,generally CH2CI2, to afford III-2. The methyl ester of III-2 can be hydrolyzed to afford III-3 under the general conditions described in Scheme 1 for the conversion W O 97/24119 PCTrUS96/20748 of I-7 to I-8. Alternatively, III-l can be reacted with succinic anhydride in the presence of an a~plopliate base, such as triethylamine, diisopropylethylamine, or ,~ pyridine, in a neutral solvent, generally CH2Cl2, to afford III-3 directly. III-3 is converted to III-4 by reaction with the known ethyl 3-amino4-pentynoate 5 (W093/07867) under the general conditions described in Scheme I for the conversion of I-6 to I-7. Hydrolysis of the ethyl ester of III-4 to provide III-5 is accomplished according to the general conditions conditions described in ~cheme I
for the conversion of I-7 to I-8.
W O 97~4119 PCTAJS96/20748 SchemP IV
NH2 N~--C02CH3 ~NH2 a,b ~--NH c N ~ C02H
~NH
Il . 0 11 H2N ~ CO2Et d ~ Boc'N ~ H''''~,'C02Et e ~
H2N~HN~CO2E~ HX f N ~ ~ N ~J'--H CO2Et ~NH O
~ Il --N~ H
a) methyl 4-(chloroformyl)butyrate, Et3N, THF; b) AcOH; c) 1.0 N NaOH, MeOH;
d) Boc-Gly, EDC, H013t ~ H,O, (i-Pr)~NEt, CH3CN; e~ TFA, CH,CI2; f) 3, EDC, HOBt ~ H20, (i-Pr)2NEt, CH3CN; g) 1.0 N LiOH, THF, H20.
The synthesis of IV-8 is acc~-mpli~h~l by the reaction of two separately prepared interm~ tes, IV-3 and IV-6. The preparation of intP.rmP.rli~h? IV-3 begins - ~6 -W O 97/24119 PCTnUS96/20748 with colllll,er-;ially available 2,3-diaminopyridine (IV-l). Accordinf to this sch~mP, IV-1 is acylated with methyl 4-(choloroformyl)butyrate in the presence of a suitable acid scavenger, such as triethylamine, diisoL,loL\ylethylarnine, or pyridine, in a neutral solvent, generally CH2Cl2 or THF, to afford an interm~ t~ monoacylated - S derivative. This delivativ~ is then cyclized, for example using r~nu~ing acetic acid, - to afford IV-2. Hydrolysis of the methyl ester of IV-2 under the general conditions described in Scheme 1 for the conversion of I-7 to I-8 affords IV-3. The preparation of the intermediate IV-6 begins with the coupling of the known ethyl 3-amino-4-pentynoate (W093/07867) with culll.llelcially available tert-butoxycarbonylglycine (Boc-Gly) under standard peptide bond forming conditions described in the previously referenced Bodansky publication, and in Scheme 1 for the conve,~ion of I-6 to I-7. The product of this reaction, IV-5, is deprotected to IV-6 under acidic conditions which are known to effect removal of a Boc protecting group. Such conditions are described in the previously referenced Bodansky and Greene publications. The two interm~ t~s IV-3 and IV-6 are coupled under standard peptide coupling conditions as previously described to afford IV-7, which is hydrolyzed to IV-8 according to the general methods described in Scheme 1 for the conversion of I-7 to I-8.
W O97/24119 PCT~US96/20748 Schen~ V
,CH3 ,CH3 H~ N~
CO2CH3 Boc CO2CH3 HO2C~I N'r'~
3 g_NH 4 ,~ N ~--~--SnBu3 ~=~ NH 5 ~O2CH3 ,CH3 S a) (Boc)20, DMAP, CH3CN; b) isobutylchloroformate, Et3N, THF, then 1,2- phenylen~ mine, then AcOH; c) (n-Bu3Sn)2, (PPh3)2PdCl2, DMF; d) CuI, (PPh3)2PdCI2, DMF; e) 4 M HCI/dioxane; f) 1.0 N NaOH, MeOH.
The synthesis of V-7 is accomplished by the reaction of two separately prepared interme~ tt~s, V-2 and V-5. V-2 is conveniently prepared by reaction ofthe readily available V-l with di-tert-butyl dicarbonate ((Boc)2O) in the presence of an acylation catalyst, preferably 4-dimethylaminopyridine (DMAP) or 4-- 5 pyrrolidinopyridine, in a neutral solvent, for example CE~3CN, THF, or CH2Cl2. The preparation of interm~ te V-5 begins with colll,ne,l~;ially available 3-iodobenzoic acid (V-3), which is converted to the ben7imi~1~7- 1e derivative V-4. According to this scheme, V-3 is reacted with isobutyl chloroformate in the presence of a suitable amine base, such as triethylamine, diisopropylethylamine, or 4-methylmorpholine,in a neutral solvent, generally CH2Cl2 or THF, to afford an interm~ te mixed anhydride delivalivt;. Without isolation, this derivative is reacted with an a~ pliate pheny!çne~ mine to produce a mono-N-acylated phenylen~ mine inferm/ ~ te. This int~ rm~ t~ is then cyclized to V-4 using acetic acid. Reaction of V-4 with bis(tributyltin) to produce V-5 occurs under palladium catalysis using, for example bis(triphenylphosphine)p~ m (II) chloride ((PPh3)2PdCl2), in an inert solvent, usually DMF. Stille-type coupling of V-2 with V-5 to afford V-6 is m~ ted by a p~ m catalyst, for instance bis(triphenylphosphine)p~ lm (II) chloride ((PPh3)2PdCl2), in the presence of copper (I) iodide (CuI), in a suitable neutral solvent, generally DMF. To obtain V-7, the protecting groups of V-6 are removed by well-established methods known to those of skill in the art and described in the previously cited Greene publication. Thus, the Boc protecting group is removed under acidic conditions, such as 4 M HCl in dioxane or TFA in CH2Cl2, and the methyl ester is hydrolyzed as generally described in Scheme 1 for the conversion of I-7 to I-8.
W O 97/24119 PCT~US96/20748 Scheme VI
CH, HO~, Boc COzCH3 Boc C02CH3 ,CH3 HO ~N
Boc CO2CH3 HO2C ~ = O
N~
Boc CO2CH3 ,CH3 Boc CO2CH3 S a) 3-butyne-1-ol, (PPh3)2PdCl2, PPh3, CuI, Et3N; b) H2, 10% Pd/C, EtOH; c) 2,2,6,6-tetramethyl-oxopiperitlinil-m chloride, CH2Cl2, then NaCl02, Na2HPO37 2-rnethyl-2-butene, H20; d) isobutyl chlc: lvfo~ ate, Et3N, then 1 ,2-phenylene~ mint-" thenAcOH; e) 1.0 N LiOH, THF, H20; f~ TFA, CH2Cl2.
CA 0224l633 l998-06-26 W O 97/24119 PCTrUS96/20748 Compound VI-1, the ~ al~lion of which is described in Scheme V, is reacted with 3-butyne-1-ol in the presence of a catalytic amount of a p~ flinm salt, usually bis(triphenylphosphine)palladium (II) chloride ((PPh3)2PdCl~), together with a catalytic amount of copper (I) iodide (CuI), in an amine solvent, such as S triethylamine (Et3N), to afford VI-2. A phosphine ligand, such as triphenylphosphine (PPh3), can be added to improve the efficiency of the reaction.
Reduction of the acetylenic unit of VI-2 is accomplished under standard hydrogenation conditions which are well-known to those of skill in the art. The rçs-llting compound, VI-3, is oxididized to the corresponding carboxylic acid VI-4, by the two-step method described by Wovkulich (J. Org. Chem. 1993, 58, 832-839).Many alternative m~thofl~ for the oxidation of a primary alcohol to the corresponding carboxylic acid have been described, and can be found in such reference volumes as "Compendium of Organic Synthetic Methods" (published by Wiley-Interscience). Conversion of the carboxylic acid of VI-4 to the ben7imid:~701e derivative VI-5 follows the procedures described in the above Schemes. The methyl ester of VI-5 is removed as described in the above Schemes, and the Boc protecting group is removed under acidic conditions, such as 4 M HClin dioxane or TFA in CH2Cl2, to afford VI-6.
CA 0224l633 l998-06-26 W O 97/24119 PCTrUS96/20748 ~r.hPme VII
HOzC--~ ~ N ~ c ~=~ NH
N ~ ~ N ~ ~
NTs ~ NTs \~/ 3 \~/ 4 N ~ NOH ~ ~_< N ~NOH
~=~ NTs ~ NTs 6 N--O N--O
N ~CO2tBu h N~co2H
NTs ~ NTs ~;~N~CO2Et ~ CO2H
a) l,2-pheny1enP~ mine, DCC, DMF, CH2Cl2; b) AcOH, THF; c) TsCl, NaH, THF;
S d) 03, CH2Cl2, MeOH, then DMS; e) NH20H ~ HCl, NaOAc, MeOH; f) NCS, DMF;
g) tert-butyl 3-butenoate, Et3N; h) 4 M HCI/dioxane, CH2Cl2; i) ethyl 3-aminobutyrate, EDC, HOBt H2O, (i-Pr)2NEt, CH3CN; j) l.0 N LiOH, THF, H2O.
W O 97/24119 PCTrUS96/20748 Commercially available 4-pentenoic acid (VII-l) is converted to the bPn7imi~1~701e derivative VII -2 using to the general procedures described ,~ previously. Protection of one of the nitrogen atoms of the ben7.imid~7ole moiety in VII -2 can be accomplished by reaction with a sulfonyl chloride, for in~t~n~e p-5 toluenesulfonyl chloride, in the presence of a suitable base, generally sodium- hydride or an aqueous alkali metal hydroxide, in an inert solvent, preferably THF, to afford VII -3. ~Itern~tive protecting groups known to those of skill in the art may be used, as long as they are co~ ible with the subsequent chemistry and can be removed when desired. Such protecting groups are described in Greene, "Protective Groups in Organic Synthesis" (published by Wiley-Interscience). Oxidative cleavage of the olefin of VII -3 to afford the aldehyde VII -4 can be conveniently accomplished by ozonolysis in an inert solvent, usually CH2Cl2 or a ll~ Ul'~ of CH2Cl2 and MeOH, followed by in-situ reduction of the ozonide with a suitable reducing agent, generally dimethylsulfide (DMS) or triphenylphosphine. ~ltr.rn~tive mPthocl~ for oxidative cleavage, such as the Lemieux-Johnson reaction (J. Org.
Chem. 1956, 21, 478) can also be used. The aldehyde is converted to the aldoximeVII -5 by standard procedures known to those of skill in the art, and this aldoxime is oxi~li7ed to the oximinoyl chloride derivative VII -6 by the methods described in WO 95tl4682 and WO 95/14683. Reaction of VII -6 with an olefin, such as tert-butyl 3-butenoate (Tet. Lett. 198~;, 26, 381 -384), in the presence of a suitable base, for in.~t~n~e triethylamine or diusopropylethylarnine, in an inert solvent such as benzene or toluene, according to the protocol described in WO 95/14682 and WO
95/14683, gives the cycloadduct I-7. The tert-butyl ester of VII -7 is removed under standard acidic conditions, generally TFA in CH2Cl2 or HCl in dioxane, to give the carboxylic acid VII -8. The carboxylic acid is activated using, for example, EDCand HOBt, or SOCl2, and the activated form is subsequently reacted with an a~prupliate amine, for in.~tAn~e a suitable derivative of ~ nin~, in a neutral solvent, such as DMF, CH2Cl2, or CH3CN, to afford VII -9. Depending on whether acid neutralization is required, an added base, such as diisopropylethylamine ((i-Pr)2NEt) or pyridine, may be used. Many additional methods for converting a W O 97/24119 PCTrUS96/20748 carboxylic acid to an amide are known, and can be found in standard reference books, such as "Compendium of Organic Synthetic Methods", Vol. I - VI (publishedby Wiley-Interscience), or Bodansky, "The Practice of Peptide Synthesis" (published by Springer-Verlag). Derivatives of ,13--alanine are readily available in either racemic 5 or optically pure form by a variety of methods known to those of skill in the art. A
reprçsent~tive method is described in WO 93/07867. The ethyl ester and sulfonyl protecting groups of VII -9 are removed using aqueous base, for example, LiOH inaqueous THF or NaOH in aqueous methanol or ethanol. The interm~ tf .
carboxylate salt is acidified with a suitable acid, for in.ct~nc~.e TFA or HCl, to afford 10 the carboxylic acid VII -10. Alternatively, the int~ te~ carboxylate salt can be isolated, if desired, or a carboxylate salt of the free carboxylic acid can be prepared by methods well-known to those of skill in the art.
Scheme vm I'IH, I ~N ~0 ~CO2Bn ~ ~CO2Bn ~ CO2Bn ,N ~o O CO2Bn CH~f ~C~, ~
~N ~ N ~f ~
S a) COCl2 in toluene, Na2CO3, H20; b) ,B-alanine benzyl ester tosylate, DMAP, - pyridine; c) CH3I, 2,6-lllti~1inç, DMF; d) BrCH2COBr, Et3N, CH2Cl2; e) NaH, DMF, f) CO, (Ph3P)2PdCl2, DIEA, 2-(methylaminomethyl)ben7.imi~1~701e dihydrochloride,NMP; g) H2, Pd/C, EtOH.
W O97/24119 PCTrUS96/20748 Following the procedures of US 5403836 and WO 9504057, except starting from 2-amino-4-iodobenzoic acid (VIII-1) rather than 2-amino-5-iodobenzoic acid,compound VIII -5 is prepared. Reaction of VIII -5 with an ~plopliate amine, for inct~nre 2-(methyl~minom~ thyl)benzimidazole, in a carbon monoxide atmosphere, 5 in the presence of a p~ rlil~m catalyst, preferably (Ph3P)2PdCl2, in an inert solvent, optimally 1-methyl-2-pyrrolidinone (NMP) gives the amide VIII -6. Depending on whether acid neutralization is required, an added base, such as diisopropylethylamine ~DIEA) or pyridine, may be used. The benzyl ester of VIII -6 is removed to afford VIII -7 under standard hydrogenolysis conditions well-known10 to those of skill in the art. ~lt~rn:~tively, the benzyl ester can be saponified using aqueous base, for example, LiOH in aqueous THF, or NaOH in aqueous methanol or ethanol. The intern~ te carboxylate salt is acidified with a suitable acid, for instance TFA or HCl, to afford the carboxylic acid. If desired, the intern~ t~-carboxylate salt of VIII -7 can be isolated, or a suitable salt of the carboxylic acid 15 can be prepared by methods well-known to those of skill in the art.
-6~-Scheme TX
O O
,~~ a ~ ~CO2Et ~~CO2Et ~[~CO2Et ~CO2Et ~ ~CO2Et SMe ¦ N
7 ~
S a) ,B-alanine ethyl ester hydrochloride, DMAP, pyridine; b) BrCH2COBr, Et3N, CH2Cl2; c) NaH, DMF; d) Lawesson's reagent, THF, 50~ C; e) CH3I, (n-Bu)4NHSO4, NaOH, CH2Cll, H20; f~ ~loL)al~ylalI~ine, pyridine HCl, toluene; g) CO, (Ph3P)lPdCl2, DIEA, 2-(methylaminomethyl)benzimidazole dihydrochloride, NMP;
h) LiOH, THF, H20.
-6~ -W O 97124119 PCT~US96/20748 Following the procedures of US 5403836 and WO 9504057, except starting from 4-iodoisatoic anhydride (IX-1, see Scheme I) rather than 5-iodoisatoic anhydride, compound IX-6 is prepared. IX-6 is converted to IX-7 following the procedures described in Scheme XIII for the conversion of XIII -5 to XIII -7.
W O 97/24119 PCTrUS96/20748 SchP,rnP. X
- BocN ~
~, N b 02H l~N~CO2Et HN~
~, N ~
~N~CO2Et ~
H OH H Cl ~N
H N ~ e N ~
N ~ CO2Et ~N
H N
l~, N ~
7 ~ N ~ C02H
., S a) l-Boc-~i~e,azille, NaBH3CN, HCl, MeOH; b) 4 M HCl/dioxane, CH2Cl2; e) SOCl2, CH2Cl2; d) 3, DIEA, DMF; e) 1.0 N NaOH, MeOH.
W O97/24119 PCTrUS96/20748 Reductive amination of the readily available 1-(ethoxycarbonylmethyl)~
piperidone (~-1, EPA 0 542 363 A2) with commercially available l-Boc-pi~e~ le and an ~ p.iate reducing agent, preferably sodium cyanoborohydride, affords amine X-2. The reaction is generally conducted under acidic catalysis, usually with S HCl, in a hydroxylic solvent, for in~tzln~e methanol or ethanol. The Boc protecting group is removed under acidic conditions, preferably HCI/dioxane or TFA, in a suitable solvent, such as CH2Cl2, to the give amine X-3. This reacts with 2-(2-chloroethyl)ben~illlida_ole (X-~;) in the presence of an ~ liate acid scavenger, for instance diisopropylethylamine (DIEA), in a polar solvent, preferably DMF, to give the coupled product X-6. 2-(2-Chloroethyl)bt~.n7;n.icl~7Qle can be preparedfrom 2-(2-hydroxyethyl)b~n7imid~7- 1e by reaction with a suitable halogenating reagent, such as thionyl chloride, or carbon tetrachloride in the presence of triphenylphosphine, in an inert solvent, for example CH2Cl2. The ethyl ester of X-6 is removed using aqueous base, for example, LiOH in aqueous THF or NaOH in 15 aqueous methanol or ethanol. The int~rm~ te~ carboxylate salt is acidified with a suitable acid, for in.stzlnce TFA or HCl, to afford the carboxylic acid X-7.
.~lt~rn~tively, the interTn~ te~ carboxylate salt can be isolated, if desired, or a carboxylate salt of the free carboxylic acid can be prepared by methods well-known to those of skill in the art.
-7~ -Sch~me XI
HN~
~, N ~f ~
~C02tBu OH
N~
NH ~N~
2 CO2tBu OH
--N
NH ~N~
3 ~,CO2H
OH
S a) 2-(3-bromopropyl)bç~7.imi~ 7.ole, DIEA, DMF, b) 4 M HClldioxane, CH2CI2.
The readily available piperazine derivative Xll-l (EPA 0 537 980 Al) is reacted with the readily available 2-(3-bromopropyl)benzimidazole (J. Org. Chem.1962, 27, 2165) in the presence of an a~ ,pliate acid scavenger, for instance 10 diisopropylethylamine (DIEA), in a polar solvent, preferably DMF, to give tnecoupled product XI-2. The tert-butyl ester protecting group is removed under standard acidic conditions, preferably HCl/dioxane or TFA, in a suitable solvent, such as CH2Cl2, to the carboxylic acid XI-3. If desired an a~r~,l)liate salt of the carboxylic acid can be prepared by methods well-known to those of skill in the art.
,.
~ -7~-W O 97/24119 PCTrUS96120748 Sch~.mP.XTT
o ~ N N ~ "" ~ OC 3 b,c ~ ~N~ NH~~O-Bn O O O
NH~N~",~NH ~OH
O O O
S a) 2-(benzirnidazolyl)propionic acid, BOP-Cl, NMM, CH2Cl2; b) LiOH, THF, H20;
c) benzyl ,13-~l~nin~tP, EDC, HOBt H20, NMM, CH2Cl2; d) H2, 10% Pd/C, AcOH, THF, H2O.
The procedures of Beavers et. al., WO 95/25091, Exam~?le 1, were followed to give XII-4, except (2-b~n7imicl~701yl)propionic acid was substituted for Na-Boc-D-lys(Cbz)-OH.
CA 0224l633 l998-06-26 W O97/24119 PCTrUS96/20748 Scheme XIII
~I O
Br ~ CO2CH3 N =< C~2CHs ~3~NH
o ~ C~2H
=~N--W c ~NH
=~ N~ ~ C02CH3 ~NH 4 =~ N~ N ~ CO2H
,~NH 5 a) 2-(arninomethyl)ben7.tmicl:~7.Qle, Et3N, benzene; b) 1.0 N LiOH, MeOH, H20; c) ,13-alanine ethyl ester, BOP, Et3N, CH3CN.
W O 97/24119 PCTrUS96/20748 A suitably functil-n~li7P.d arnine, such as 2-(aminomethyl)ben7imi(l;~7ole~ is reacted with dimethyl 4-bromomethylbenzene-1,3-dicarboxylate (XIII-l;
.y~ esi~ as in EP 0540334Al) under the general conditions described for the ~,c~dLion of l-H-isoindole-5-carb~-x~mitl~7 2,3-dihydro-N-(2-carboxy-ethyl)-2-[2-S (piperidinyl)ethyl]-3-oxo (preparation 1-12, EPA 0 540 334 Al), to afford XIII -2.
The methyl ester of XIII -2 is hydrolyzed using aqueous base, for example, LiOH in aqueous THF or NaOH in aqueous methanol or ethanol, and the int.orm~ t~
carboxylate salt is ~ci~lifi~.-l with a suitable acid, for instance TFA or HCl, to afford the carboxylic acid XIII -3. The carboxylic acid of XIII -3 is converted to an activated form of the carboxylic acid using, for example, EDC and HOBt, SOC12, or BOP reagent, and the activated form is subsequently reacted with an a~ iate amine, for in~t:~nre ,B-alanine ethyl ester, in a suitable solvent such as DMF, CH2Cl2, or CH3CN, to afford XIII -4. Depending on whether acid neutralization is required, an added base, such as diisopropylethylamine ((i-Pr)2NEt) or pyridine, may be used.
Many additional methods for converting a carboxylic acid to an amide are known, ~ and can be found in standard reference books, such as "Compendium of Organic Synthetic Methods", Vol. I - VI (published by Wiley-Interscience), or Bodansky, "The Practice of Peptide Synthesis" ~published by Springer-Verlag). Ester hydrolysis as described above for the conversion of XIII -2 to XIII -3 then affords XIII -S. Alternatively, the int~rm.o~ii~tf~. carboxylate salt of XIII -~ can be isolated, if desired, or a carboxylate salt of the free carboxylic acid can be prepared by methods well-known to those of skill in the art.
CA 0224l633 l998-06-26 PCTrUS96/20748 W O 97/2~119 Scheme XIV
NH~ ~OH N' ~3~OH
'~3'o CO~BnCH~'O CO2Bn d N ~N~
~NH O O
\G/ 5 CO2Bn N ~ ICH~3~
~q~NH O O
\~/ 6 CO2H
a) (Boc)20, NaOH, 1,4-dioxane, HlO; b) BrCH2COlBn, K2C03, acetone; c) 4 M
HCI/dioxane; d) 2-(ben7.imi~1~7.olyl)acetic acid, EDC, DIEA, DMF; e) H2, 5 % Pd/C, MeOH.
XIV-1 is treated with di-tert-butyl dicarbonate and sodium hydroxide in 10 aqueous dioxane to afford XIV -2, which is alkylated on oxygen with benzyl - bromo~f.et~te and potassium carbonate in acetone to give XIV -3. The Boc group in -7~-XIV -3 is removed with hydrogen chloride in dioxane, and the rçsllltin~ XIV -4 is acylated on nitrogen with 29ben7imi-1~7Qlyl)acetic acid, EDC and DIEA in DMF to give XIV -5. The benzyl ester in XIV -5 is cleaved by tre~tml-.nt with hydrogen and Fz~ linm-on-carbon in methanol to give XIV -6.
S
Sch.ome XV
H2N ~ NH ~O
al NH ~ NH~ NH ~f O J3 O O
b NH ~ NH~ NH ~OH
O O
a) 2-(benzirnidazolyl)acetic acid, EDC, DIEA, DMF; b) NaOH, H20, CH30H.
XI-l, prepared as described in Alig et. al., EPA 0372486, is con~ n~e~ with a suitable substituted carboxylic acid, such (2-ben7imitl~701yl)acetic acid, in the 15 presence of EDC and DIEA, and in a suitable solvent, e.g., DMF or acetonitrile.
-7~ -W O97124119 PCT~US96/20748 Many additional methods for converting a carboxylic acid to an amide are known, and can be found in standard reference books, such as "Compendium of Organic ,. Synthesis". Vol. I-VI (published by Springer-Verlag). Hydrolysis of the ester is accomplished by saponirlcation with a suitable reagent, e.g., sodium hydroxide, in a 5 suitable solvent, e.g., aqueous methanol. AAternatively, a benzyl ester may be- converted to the acid by tre~tmfA.nt with a suitable catalyst, e.g., Pd/C, and hydrogen in a suitable solvent, e.g., methanol, ethanol or acetic acid.
Schem~ XVI
1~
~[3~OH
H2N ~ N3 O ~<0 al 1 O-tBu ~3 0 ,<o 2 O-tBu b l ~OH
Q--N o ~W
NH ~W--NH~N~} --COzH
a) 2-(benzill,Adazolyl)acetic acid, EDC, DIEA, DMF; b) TFA
CA 0224l633 l998-06-26 XVI-l, prepared as described in Alig et. al., EPA 0505868, is condensed with a suitable substituted carboxylic acid, such (2-ben7imi~1~7Olyl)acetic acid, in the presence of EDC and DIEA, in a suitable solvent, e.g., DMF or acetonitrile, to give XVI -2. Many additional methods for converting a carboxylic acid to an amide are5 known, and can be found in standard reference books, such as "Compendium of Organic Synthesis". Vol. I-VI (published by Springer-Verlag). Hydrolysis of the ester in XVI -2 is accomplished with trifluoroacetic acid or hydrogen chloride to give XVI -3. Alt~rn~tively, the ester in XVI -2 may be saponified with a suitable reagent, e.g., lN NaOH, in a suitable solvent, e.g., methanol.
Seht-mt- XVII
~IL N N ~ O-tBu H2N 0~O t a l ~0 b l ~ ~ A co H
a) 2-(b~n7imi-1~701yl)acetic acid, EDC, DIEA, DMI;'; b) TFA, CH2Cl2.
W O 97/24119 PCT~US96/20748 XVII-1, prepared as described in Sugihara, et. al., EP 0529858, is condensed with a suitable substituted carboxylic acid, such as (2-bell7imi~1~701yl)acetic acid, to " give XVII -2, and the tert-butyl ester is cleaved with TFA, following the general procedure of Sugihara, et. al., Example 59, to give XVII -3. Many additional - 5 methods for converting a carboxylic acid to an amide are known, and can be found ~ in standard reference books, such as "Compe~-lium of Organic Synthesis", Vol. I-VI
(published by Springer-Verlag).
Schem~ XVITI
Ms-o Q--N
~~~0 a ~ 2 4~ 0 0--t-Bu O-tBu ~0 3 "~0 OH
a) 4-r2-(benzimidazolyl)methyl]phenol, Cs2CO3, DMF; b) TFA
Compound XVIII-1, prepared as described in Himmelsbach, et. al., Australian Patent Application AU-A-86926/91, Example VI~28), is treated with a suitable ~ub~ uled phenol, such as 4-[2-(benzimidazolyl)methyl]phenol, prepared W O 97/24119 PCTrUS96/20748 by the general procedure of Wahlgren and Addison, J. Heterocycl. Chem., 1989, 26, 541-3, following the general method of Himmelsbach, et. al., Example 3(51), to give XVIII -2. The tert-butyl ester in XVIII -2 is hydrolyzed with lN NaOH in CH30H .
following the general procedure of Himmelsbach, Example 7(3) to give XVIII -3.
5 Alternatively, the tert-butyl ester may be cleaved with TFA or HCl.
EP 0567968, Nov. 3, 1993, Linz, G., et al.: Describes amidinobiphenyl-lactam-acetic acid and amidinophenyll~f t~mphenylpropionic acid analogs.
EP 0574808, June 11, 1993, Pieper, H., et al.: Describes amidinobiphenyl-X-acetic acid ester analogs.
Der 93-406657/51, Austel, V.., et al.: Describes amidinobiphenyl analogs.
EP 587134, (Der 94-085077/11) Mar. 16, 1994, Himmerlsbach, F. D. D., et al., Describes amidinophenyltriazolone analogs.
EP 589874, Apr. 6, 1994, Grell, W., et al.
~P534005), DE 4234295, Apr. 14, 1994, Pieper, H., et al., Describes heteroaryl-azacyclohexylcarboxylic acid analogs.
EP 0592949, Apr. 20, 1994, Pieper, H. D., et al., Describes arnidinophenyl-4piperi-1in~mi~1o-4-cyclohexylcarboxylic acid analogs.
EP 596326, May, 11, 1994, Maier, R. et al.
DE 4241632, June 15, 1994, Himmelsbach, F., et al., Describes piperidinophenylamido-phenyl~lvL,ionyl analogs.
EP 0604800 A, Jul. 6, 1994, Himmelsbach, F. et al., Describes piperidinophenylamido-phenyl~l~nine derivatives.
DE 4302051, (Der 94-235999/29) July, 28, 1994, describes compounds con~inin~ a 2H-pyrazol-5-one.
EP 0608858 A, Aug, 3, 1994, Linz, G. D., et al., Describes amidino-biphenyl compounds.
DE 4304650, (Der 94-256165/32), Aug, 18, 1994, Austel, V., et al., describes compounds with a 5,6 template.
- 30 EP 611660, Aug, 24, 1994, Austel, V., et al., Describes tricyclic template.
W O97/24119 PCT~US96~0748 DE 4305388, (Der 94-264904/33), Aug. 25, 1994, I~Timm~l~b~rh~ F., et al., Describes 6,6 and 7,6 templates.
(P5D4005), EP 612741, (Der 94-265886/33), Aug. 31, 1994, Himmelsbach, F., et al., Describes 6,6 and 7,6 templates.
EP 0639575 A, Feb. 22, 1995, Linz, G., et al.: Describes tetrahydrothiazolo-[5,4,c]pyridine cation replacements.
DE 4324580, Jan. 26, 1995, Linz, G. et al.
EP 0638553, Feb. 15, 1995, Himm~l~b~h, F., et al.
F. rTiumm~olsbach, V. Austel, G. Kruger, H. Pieper, H. Weisenberger, T. H. Muller, and W. G. Eisert, in XIIth Int. Symp. on Med. Chem. Basel, Book of Abstracts, 47, 1992.
V. Austel, W. Eisert, F. Himmelsbach, G. Kruger, G. Linz, T. Muller, H. Pieper, and J. Weisenberger, Natl. Mtg. Amer. Chem. Soc. Book of Abstracts, Denver, Div. Med. Chem., 1993.
Muller, T. H.; Schurer, H.; Waldmann, L.; Bauer, E.; ~immPlsbach, F.; Binder, K., Orally Activity of BIBU 104, a Prodrug of the Non-peptide Fibrinogen Receptor Antagonist BIBU 52, in Mice and Monkeys, Thromb. Haem., 69, 975, 1993.
Univ. California WO 94/14848, July, 7, 1994, Zanetti, M. RGD peptides from CDR.
Univ. New York WO 94/00144, June 29, 1993, Ojima, I. et al.: Describes RGD peptide mllltimtors.
Yeda lRes. and Dev. Co.
WO 93/09795, (Der 93-182236/22), Lido, O. et al.: Describes Gn~ni~inopentanoic acid analogs.
7PnP(~ -WO 9422834, Oct. 13, 1994, Wayne, M. G., et al. Describes pyridinopiperazino-phenylcarbonyl-arnino acids.
WO 9422835, Oct. 13, 1994, Wayne, M. G., et al. Describes pyridinopiperidino-amidophenylacetic acids.
5EP 632016, Jan. 4, 1995, Brewster, A. G.. , et al. Describes - pyridinopropionylhyd,d~i"ylbenzoyl analogs.
EO 632019, Jan. 4, 1995, Brown, G., Shute, R. E.
EO 632020, ~an. 4, 1995, Brown, G., Shute, R. E.
~n cases wherein the compounds of this invention may have one or more chiral centers, unless specified, this invention includes each unique nonracemiccompound which may be synthesized and resolved by conventional techniques. In cases in which compounds have unsaturated carbon-carbon double bonds, both the cis (Z) and trans (E) isomers are within the scope of this invention. The m~z~nin~ of 15 any substituent at any one occurrence is independent of its m~ning, or any other substituent's m~ning, at any other occurrence.
Abbreviations and symbols commonly used in the peptide and chemical arts are used herein to describe the compounds of this invention. In general, the amino acid abbreviations follow the IUPAC-IUB Joint Commission on Biochemical 20 Nomenclature as described in Eur. J. Biochem., 158, 9 (1984).
Cl 4alkyl as applied herein means an optionally substituted alkyl group of 1 to 4 carbon atoms, and inc.lucles methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl and t-butyl. Cl 6alkyl additionally includes pentyl, n-pentyl, isopentyl, neopentyl and hexyl and the simple aliphatic isomers thereof. C0 4alkyl and C0 6alkyl 25 additionally indicates that no alkyl group need be present ~e.g., that a covalent bond is present).
Any Cl 4alkyl or Cl 6 alkyl, C2 6 alkenyl, C2 6 alkynyl or Cl 6 oxoalkyl may be optionally substituted with the group RX, which may be on any carbon atom that results in a stable structure and is available by conventional synthetic techniques.
30 Suitable groups for Rx are Cl 4alkyl, OR, SR, Cl 4alkyl, Cl 4alkylsulfonyl, W O 97/24119 PCTrUS96/20748 C I 4alkylsulfoxyl, -CN, N(R )2, CH2N(R )2. -N02, -CF3, -C02R -CON(R )2~ -COR, -NR C(O)R, OH, F, Cl, Br, I, N3, or CF3S(O)~-,wherein r is O to 2.
Ar, or aryl, as applied herein, means phenyl or naphthyl, or phenyl or naphthyl substituted by one to three subctitllent~ sueh as those defined above for 5 alkyl, espeeially Cl~alkyl, Cl 4alkoxy, Cl 4alkthio, trifluoroalkyl, N3, OH, C02H
F, Cl, Br or I.
Het, or heterocyele, in~lic~tl~.s an optionally ~llb~liLuled five or six membered monoeyelic ring, or a nine or ten-membered bieyelie ring cf-nt~ining one to three heteroatoms ehosen from the group of nitrogen, oxygen and sulfur, whieh are stable 10 and available by eonventional ehPmic~l synthesis. Illustrative heterocycles are benzofuran, ben7imicl~01e, bellzoL)yl~l, benzothiophene, biotin, furan, imicl~ole, indoline, morpholine, piperidine, ~iL~t;ld~ille, pyrrole, pyrro}idine, pyridine,tetrahydropyridine, pyridine, thiazole, thiophene, quinoline, isoquinoline, and tetra-and ~lhyd~ quinoline and isoquinoline. Any ~cçc.cihle combination of up to 15 three substit~entc on the Het ring, such as those defined above for alkyl that are available by ehemieal synthesis and are stable are within the seope of this invention.
C3 7cyeloaL~yl refers to an optionally substituted carbocyelic system of three to seven earbon atoms, whieh may eontain up to two unsaturated earbon-earbon bonds. Typieal of C3 7eyeloalkyl are eyelopropyl, cyclobutyl, cyclopentyl, 20 cyelopentenyl, eyelohexyl, eyelohexenyl and eyeloheptyl. Any eombination of up to three substi~uent.~, such as those defined above for alkyl, on the cycloalkyl ring that is available by conventional chemieal synthesis and is stable, is within the seope of this invention.
When Rb and Rc are joined together to form a five- or six-membered 25 aromatie or non-aromatic carbocyclic or heterocyclie ring fused to the ring to whieh Rb and Rc are attached, the ring formed will generally be a five- or six-membered heterocycle s~lecte~ from those listed above for Het, or will be a phenyl, eyclohexyl or cyclopentyl ring. Preferably Rb and Rc will be -D1=D2-D3=D4 wherein Dl - D4 are independently CH, N or C-RX with the proviso that no more than two of D 1 - D4 W O 97/24119 PCT~US96/20748 are N. Most preferably, when Rb and Rc are joined together they form the group -CH=CH-CH=CH-.
- Certain radical groups are abbreviated herein. t-Bu refers to the tertiary butyl radical, Boc refers to the t-butyloxycarbonyl radical, Fmoc refers to the 5 fluorenylmethoxycarbonyl radical, Ph refers to the phenyl radical, Cbz refers to the - benzyloxycarbonyl radical, BrZ refers to the o-bromobenzyloxycarbonyl radical, ClZ refers to the o-chlorobenzyloxycarbonyl radical, Bzl refers to the benzyl radical, 4-MBzl refers to the 4-methyl benzyl radical, Me refers to methyl, Et refers to ethyl, Ac refers to acetyl, Alk refers to Cl 4alkyl, Nph refers to 1- or 2-naphthyl and cHex 10 refers to cyclohexyl. Tet refers to 5-tetrazolyl.
Certain reagents are abbreviated herein. DCC refers to dicyclohexylcarbo-liimi(~, DMAP refers to dimethylaminopyridine, DIEA refers to diisopropylethyl amine, EDC refers to 1-(3-dimethylaminopropyl)-3-ethylcarborliimi(le, hydrochloride. HOBt refers to l-hydroxybenzotri~7Ole, THF
15 refers to tetrahydrofuran, DIEA refers to diisopropylethylamine, DME refers to dimethoxyethane, DMF refers to dimethylform~micle, NBS refers to N-bromosuccinimide, Pd/C refers to a palladium on carbon catalyst, PPA refers to 1-propanephosphonic acid cyclic anhydride, DPPA refers to diphenylphosphoryl azide, BOP refers to benzotriazol-l-yloxy-tris(dimethyl-amino)phosphonium 20 hexafluorophosphate, HF refers to hydrofluoric acid, TEA refers to triethylamine, TFA refers to trifluoroacetic acid, PCC refers to pyridinium chlorochromate.
Compounds of the formula ~ (V) are prepared, for example, by reacting a compound of formula (XIX) with a compound of formula (XX), wherein Ll and L2 are groups which may react to fo;m a covalent bond in the moiety W, by methods 25 generally known in the art.
~ ~> L~ > W-A
RC N + Ll-A RC N
(XIX) (XX) (I) Typical methods include coupling to form amide bonds, nucleophilic displ~rçm~nt reactions and p~ m catalyzed couplings. For instance, when W
W O 97/24119 PCTrUS96/20748 contains an ether or amine linkage, the bond may be formed by a displacement reaction, and one of Ll and L2 will contain an amino or hydroxy group and the other will contain a displaceable group, such as a chloro, bromo or iodo group. When Wcontains an amide bond, typically one of Ll and L2 will contain an amino group, and 5 the other will contain a carboxylic acid group. In another approach, Ll may be an aryl or heteroaryl bromide, iodide or trifluoromethylsulfonyloxy derivative and L2 may contain an amino group and the amide linkage may be formed by p~ m-catalyzed aminocarbonylation with carbon monoxide in a suitable solvent such as dimethylform~micle or toluene.
It will be apparent that the precise identity of Ll and L2 will be dependent upon the site of the linkage being }'ormed. General methods for preparing the linkage -(CHR")~-U-(CHR")s-V- are described, for example, in EP-A 0 372 486 and EP-A 0 381 033 and LP-A 0 478 363, which are incol~oldt~d herein by reference.
For in~t~nre, if V is CONH, Ll may be -NH2, L2 may be O~I (as in an acid) 15 or Cl (as in an acid chloride), and R6 may be W-(CRr2)q-Z-(CR'Rl0)r~U-(CRI2) C(O), with any functional groups optionally protected. For example, R6 may be (benzyloxycarbonyl-amidino)benzoyl- or (N~Boc,NgUan-Tos)arginyl-. When L2 is OH, a coupling agent is used.
Similarly, if V is NHCO, Ll may be -CO2H or CO-Cl, L2 may be -NH2, and R6 may be W~(CR12)q~Z(CRIRl~)r-u-(cRl2)s-. For example, R6 may be (benzyloxycarbonyl-amidino)phenyl, (benzyloxycarbonylamino)methylbenzyl- or 6-(benzyloxycarbonylamino)hexyl- .
Where V is NHSO2, Ll may be SO2Cl, L2 may be -NH2 and R6 may be as above. Where V is SO2NH, Ll may be -NH2 and L2 may be SO2CI. Methods to 25 prepare such sulfonyl chlorides are disclosed, for in~t~n~e, in J. Org. ~he~l., 23, 1257 (1958).
If V is CH=CH, Ll may be -CHO, L2 may be CH=P-Ph3 and R6 may be W-(CR12)q-Z-(CRlRlO)~U-(CRl2)s-- ~l~rnzlt~ly, Ll may be CH=P-Ph3, L2 may be CHO, e.g., R6 may be W~(CR~2)q~Z~(CRlRl0)r-u-(cRl2)s-l-cHo.
-~6 -~ .
W O97/24119 PCTrUS96/20748 Where V is CH2CH2 may be obtained by reduction of a suitably protected compound wherein V is CH=C~I.
Where V is CH20, CH2N or C--C, L1 may be -OH, -NH or -C~ C H,respectively; L2 may be -Br; and R6 may be W~(CR~2)q~Z(CR~RI~)r~u-(cR~2)s-~ For example, R6 may be (benzyloxycarbonylarnino)-methylbenzyl- or 2-(N-benzyl-4-piperidinyl)-ethyl.
Similarly where U or V is OCH2, NR'CH2 or C~ C, Ll may be -CH2Br and L2 may be -OH, -NH or -C= C H, le~.~e.;lively. Alternately, when U or V is C--C, Ll maybe Br, I or CF3SO3, L2 may be C--C H and the coupling may be catalyzed by p~ m and a base.
Compounds wherein V is CHOHCH2 may be prepared from a suitably protected compound where V is CH=CH by the procedure disclosed in J. Org.
Chem., 54, 1354 (1989).
Compounds wherein V is CH2CHOH may be obtained from a suitably protected compound where V is CH=CH by hydroboration and basic oxidation as disclosed in Tet. Lett., 31, 231 (1990).
The core 6-7 fused ring system is prepared of formula (VI) by methods well known in the art, e.g., Hynes, et al., J. Het. Chem., 1988, 25, 1173; Muller, et al., Helv. Chim. Acta., 1982, 65, 2118; Mori, et al., Heterocycles, 1981, 16, 1491.
Similarly, methods for preparing benzazepines, 1,4-benzofhi~7.e.pin~ s, 1,4-benzoxazepines and 1,4-benzodiazepines are known and are disclosed, for in.~t~n~e, in Bondinell, et al., International Patent Application WO 93/00095.
The sl~htom~s which follow detail the ~l~dLion of the compounds of the instant invention.
W O 97/24119 PCT~US96/20748 S. ' - ~ I
CH30~ ~ CH~0~[~
o CO2Et CH30~ H0 ~
3 C02Et 4 C02Et TfO ~ HO C~
C02Et C02Et g_--~ N r ~ ~ h a) EtOAc/LiN(TMS)2, THF; b) Et3SiH, BF3 ~ OEtz, CH2Cl2; c) H2~ 10% Pd/C, EtOH;
d) EtSH, AICl3, CH2Cl2; e) Tf20, 2,6-h-ti~lin~., CH~Cl2; f) CO, KOAc, Pd(OAc)2, 5 dppf, DMSO; g) 2-(methylaminomethyl)l~e~ ole dihydrochloride, EDC, EIOBt ~ H20, (i-Pr)2NEt, CH~CN; h) 1.0 N NaOH, EtOH.
CA 0224l633 l998-06-26 An ~p,op.iately substituted deoxybenzoin, such as 2-(4-methoxyphenyl)-l-phenylethanone (Chem. Ber. 1958, 91, 755-759), is reacted in an aldol-type reaction with the enolate of ethyl acetate, which can be generated from ethyl acetate on exposure to an a~pr~p.iate amide base, for in~t~nre lithium diisopropylamide (LDA) 5 or lithium bis(trimethylsilyl)amide (LiN(TMS)2), to afford I-2. Fre~uently, THF is - the solvent of choice for an aldol reaction, although THF in the presence of various additives, for in.~t~nre HMPA or TMEDA, is often used. Reaction of I-2 with triethylsilane (Et3SiH) in the presence of boron trifluoride etherate (BF3 ~ OEt2) according to the general protocol of Orphanopoulos and Smonu (Synth. Commun.
10 1988, 833) for the reduction of tertiary benzylic alcohols affords I-3, together with the olefinic product derived from ~ limin:~tion of the alcohol. The olefinic product can be conveniently converted to I-3 by hydrogenation over a F~ m catalyst, such as palladium metal on activated carbon (Pd/C), in an ~pl~pliate inert solvent, for in~t:~nre mr.th~nol, ethanol, or ethyl acetate. Removal of the methyl ether of I-3 15 to give I-4 can be accomplished by reaction with eth~nethiol (EtSH) in the presence of a Lewis acid catalyst, preferably anhydrous ~lnminllm trichloride (AlCl3), in an inert solvent, for in~t~nce CH2Cl2. Other useful methods for removal of a methylether are described in Greene, "Protective Groups in Organic Synthesis" (published by Wiley-Interscience). Alcohol I-4 is converted to its trifluoromethanesulfonate 20 ester I-5 by reaction with trifluoromethanesulfonic anhydride (Tf2O) in the presence of a suitable non-nucleophilic amine base, such as 2,6-lutidine, in an inert solvent, generally CH2Cl2. I-~ reacts with carbon monoxide (CO) in the presence of potassium acetate, 1, l '-bis(diphenylphosphino)ferrocene (dppf), and a F~ lillmcatalyst, for in~t~nce palladium acetate (Pd(OAc)2), in a suitable solvent, preferably 25 DMSO, according to the general method described by Cacchi and Lupi (Tet. Lett.
1992, 33, 3939) for the carboxylation of aryl trifluorometh~nt-sulfonates. The resulting benzoic acid derivative I-6 is converted to an activated form of the carboxylic acid using, for example, EDC and HOBt, or SOCl2, and the activated form is subsequently reacted with an ~pl~,p.iate amine, for in~t~nre 2-30 (methylaminomethyl)methylben7.imi-l~7.ole dihydrochlnri~le, in a suitable solvent W O 97/24119 PCT~US96/20748 such as DMF, CH~Cl2, or CH3CN, to afford I-7. Df~pçn-lin~ on whether acid neutralization is required, an added base, such as diisopropylethylamine ((i-Pr)2NEt) or pyridine, may be used. Many additional methods for converting a carboxylic acid to an amide are known, and can be found in standard reference books, such as 5 "Compendium of Organic Synthetic Methods", Vol. I - VI (published by Wiley-Interscience), or Bodansky, "The Practice of Peptide Synthesis" (published by Springer-Verlag). The ethyl ester of I-7 is hydrolyzed using aqueous base, for example, LiOH in aqueous THF or NaOH in aqueous methanol or ethanol, and the intermediate carboxylate salt is acidified with a suitable acid, for instance TFA or 10 HCl, to afford the carboxylic acid I-8. ~lt~rn~tively, the interm~ t.o carboxylate salt can be isolated, if desired, or a carboxylate salt of the free carboxylic acid can be prepared by methods well-known to those of skill in the art.
W O 97/24119 PCT~US96/20748 Scheme II
~ HO2C~ t-BUO2C~0~ t-Bu02C~C02CH3 ~- 1 23 t-BuO2C ~ t-BuO2C ~ e t-Bu02C ~ t-Bu02C~,CH3) CON(CH3)2 CO2Et H~2C~ CON(CH3)2 ~ N~N~ "CH3)2 ~CO2Et (~NH CH3 CO2Et N~ N J~ CH3)2 ~ NH CH3 CO2H
a) isobutylene, TfOH, CH2C12; b) methyl acrylate, Pd(OAc)2, P(tol)3, (i-Pr)2NBt,propionitrile; c) H2, 10% Pd/C, MeOH, EtOAc; d) 1.0 N LiOH, THF, H2O; e) dimethylamine hydrochloride, EDC, HOBt ~ H20, (i-Pr)2NEt, CH3CN; f) LiN(TMS)2, - THF, then BrCH2CO2Et; g) TFA, CH2Cl2; h) 2-(methylaminomethyl)ben7.imi(1~7.ole dihydrochloride, EDC, HOBt H20, (i-Pr),NEit, CH3CN; i) 1.0 N LiOH, THF, H20.
W O 97/24119 PCTrUS96t20748 Commercially available 4-bromobenzoic acid (II-l) is converted to the tert-butyl ester II-2 by reaction with isobutylene in the presence of a catalytic arnount of an acid, such as trifluorom~thAn.osulfonic acid (TfOH) or sulfuric acid, in _n inert solvent, generally CH2Cl2 or diethyl ether. Alternative methods for the formation of 5 tert-butyl esters are described in Greene, "Protective Groups in Organic Synthesis"
(published by Wiley-Interscience). Other esters can be employed, as long as theyare compatible with the subsequent chemistry and can be removed selectively whendesired. A Heck-type reaction between II-2 and methyl acrylate affords II-3. Thegeneral conditions for the Heck reaction have been reviewed by Heck (Org.
lO Reactions 1982, 27, 345). For ~I-2, reaction with methyl acrylate in the presence of palladium ~II) acetate (Pd(OAc)2) and tri-ortho-tolylphosphine (P(tol)3) in an inert solvent, such as CH3CN, propionitrile, or toluene, in the presence of an ~p.o~,iate acid scavenger, such as diisopropylethyla~lnine ((i-Pr)2NEt), affords II-3. Reduction of the a,l3-unsaturated ester of II-3 to afford the saturated compound II-4 occurs 15 under standard hydrogenation conditions, for instance reaction with hydrogen gas in the presence of a suitable catalyst, preferably palladium metal on activated carbon (Pd/C), in an inert solvent, generally m(~thAn~l, ethanol, ethyl acetate, or mixtures thereof. The methyl ester of II-4 is hydrolyzed using aqueous base, for example,LiOH in aqueous THF or NaOH in aqueous methanol or eth_nol, and the 20 interm~-liAte carboxylate salt is acidified with a suitable acid, for in~t~n~e T~A or HCl, to afford the carboxylic acid II-5. Tnis is converted to an activated form of the carboxylic acid using, for example, EDC and HOBt, or SOC12, and the activated form is subsequently reacted with an a~3p~ iate a~nine, for in~tAnce dimethylamine hydrochloride, in a suitable solvent such as DMF, CH2CI~, or CH3CN, to afford II-6.
25 Depending on whether acid neutralization is required, an added base, such as diisopropylethylamine ((i-Pr)2NEt) or pyridine, may be used. Many additional methods for converting a carboxylic acid to an amide are known, and can be foundin standard reference books, such as "Compendium of Organic Synthetic Methods", Vol. I - VI (published by Wiley-Interscience), or Bodansky, "The Practice of Peptide 30 Synthesis" (published by Springer-Verlag). Reaction of II-6 with an amide base, for CA 0224l633 l998-06-26 W O 97/24119 PCT~US96/20748 in.~t:~nCe~ lithium bis(trimethylsilyl)amide (3_iN(TMS)2), sodium bis(trimethylsilyl)amide (NaN(TMS)2), pot~c~ m bis(trimethylsilyl)amide , (KN(TMS)2), or lithium diisopropylamide (LDA), in an inert solvent, generally THF
or ethylene glycol dimethyl ether (DME), affords an interm~ te amide enolate.
5 This is generally not isolated, but rather is reacted in situ with an electrophile, for example ethyl bromoacetate, to afford the alkylated product II-7. Various additives known to those of skill in the art, for in~t~n~e HMPA, teLlalllelhylethylen~ mine (TMEDA), or 1,3-dimethyl-3,4,5,6-tetrahydro-2(1H)-pyrimidinone (DMPU), can be used to improve the eff1ciency of the alkylation reaction. ). The tert-butyl ester 10 group of II-7 is removed under acidic conditions, generally with TFA or HCl, in an inert solvent, usually CH2Cl2, 1~4-dioxane, or l~ Lules thereof, to afford the acid II-8. Other useful methods for the removal of tert-butyl esters are described in Greene, "Protective Groups in Organic Synthesis" (published by Wiley-Interscience). II-8 is converted to II-9 under the general conditions described in Scheme I for the conversion of I-6 to I-7, and II-9 is converted to II-10 by the general conditions described in Scheme I for the conversion of I-7 to I-8.
Schem~. m ~_NH a~_NH O COzCH3 ~N CO H
~ NH N ~ ~ COzEt N ~ N ~ ~ H CO2H
g_NH O
a) 3-carbomethoxypropionyl chloride, (i-Pr)2NEt, CH2Cl2; b) 1.0 N NaOH, MeOH;
c) ethyl 3-amino-4-pentynoate, EDC, HOBt ~ H20, (i-Pr)2NEt, CH~CN, DMF; d) 1.0 N LiOH, THF, H20, CH3CN.
Readily available 2-(2-aminoethyl)be~,~.;.l.itl~7.ole is reacted with 3-1() carbomethoxy~lopionyl chloride in the presence of an al,~r.,l,liate acid scavenger, such as triethylamine, diisopropylethylamine, or pyridine, in a neutral solvent,generally CH2CI2, to afford III-2. The methyl ester of III-2 can be hydrolyzed to afford III-3 under the general conditions described in Scheme 1 for the conversion W O 97/24119 PCTrUS96/20748 of I-7 to I-8. Alternatively, III-l can be reacted with succinic anhydride in the presence of an a~plopliate base, such as triethylamine, diisopropylethylamine, or ,~ pyridine, in a neutral solvent, generally CH2Cl2, to afford III-3 directly. III-3 is converted to III-4 by reaction with the known ethyl 3-amino4-pentynoate 5 (W093/07867) under the general conditions described in Scheme I for the conversion of I-6 to I-7. Hydrolysis of the ethyl ester of III-4 to provide III-5 is accomplished according to the general conditions conditions described in ~cheme I
for the conversion of I-7 to I-8.
W O 97~4119 PCTAJS96/20748 SchemP IV
NH2 N~--C02CH3 ~NH2 a,b ~--NH c N ~ C02H
~NH
Il . 0 11 H2N ~ CO2Et d ~ Boc'N ~ H''''~,'C02Et e ~
H2N~HN~CO2E~ HX f N ~ ~ N ~J'--H CO2Et ~NH O
~ Il --N~ H
a) methyl 4-(chloroformyl)butyrate, Et3N, THF; b) AcOH; c) 1.0 N NaOH, MeOH;
d) Boc-Gly, EDC, H013t ~ H,O, (i-Pr)~NEt, CH3CN; e~ TFA, CH,CI2; f) 3, EDC, HOBt ~ H20, (i-Pr)2NEt, CH3CN; g) 1.0 N LiOH, THF, H20.
The synthesis of IV-8 is acc~-mpli~h~l by the reaction of two separately prepared interm~ tes, IV-3 and IV-6. The preparation of intP.rmP.rli~h? IV-3 begins - ~6 -W O 97/24119 PCTnUS96/20748 with colllll,er-;ially available 2,3-diaminopyridine (IV-l). Accordinf to this sch~mP, IV-1 is acylated with methyl 4-(choloroformyl)butyrate in the presence of a suitable acid scavenger, such as triethylamine, diisoL,loL\ylethylarnine, or pyridine, in a neutral solvent, generally CH2Cl2 or THF, to afford an interm~ t~ monoacylated - S derivative. This delivativ~ is then cyclized, for example using r~nu~ing acetic acid, - to afford IV-2. Hydrolysis of the methyl ester of IV-2 under the general conditions described in Scheme 1 for the conversion of I-7 to I-8 affords IV-3. The preparation of the intermediate IV-6 begins with the coupling of the known ethyl 3-amino-4-pentynoate (W093/07867) with culll.llelcially available tert-butoxycarbonylglycine (Boc-Gly) under standard peptide bond forming conditions described in the previously referenced Bodansky publication, and in Scheme 1 for the conve,~ion of I-6 to I-7. The product of this reaction, IV-5, is deprotected to IV-6 under acidic conditions which are known to effect removal of a Boc protecting group. Such conditions are described in the previously referenced Bodansky and Greene publications. The two interm~ t~s IV-3 and IV-6 are coupled under standard peptide coupling conditions as previously described to afford IV-7, which is hydrolyzed to IV-8 according to the general methods described in Scheme 1 for the conversion of I-7 to I-8.
W O97/24119 PCT~US96/20748 Schen~ V
,CH3 ,CH3 H~ N~
CO2CH3 Boc CO2CH3 HO2C~I N'r'~
3 g_NH 4 ,~ N ~--~--SnBu3 ~=~ NH 5 ~O2CH3 ,CH3 S a) (Boc)20, DMAP, CH3CN; b) isobutylchloroformate, Et3N, THF, then 1,2- phenylen~ mine, then AcOH; c) (n-Bu3Sn)2, (PPh3)2PdCl2, DMF; d) CuI, (PPh3)2PdCI2, DMF; e) 4 M HCI/dioxane; f) 1.0 N NaOH, MeOH.
The synthesis of V-7 is accomplished by the reaction of two separately prepared interme~ tt~s, V-2 and V-5. V-2 is conveniently prepared by reaction ofthe readily available V-l with di-tert-butyl dicarbonate ((Boc)2O) in the presence of an acylation catalyst, preferably 4-dimethylaminopyridine (DMAP) or 4-- 5 pyrrolidinopyridine, in a neutral solvent, for example CE~3CN, THF, or CH2Cl2. The preparation of interm~ te V-5 begins with colll,ne,l~;ially available 3-iodobenzoic acid (V-3), which is converted to the ben7imi~1~7- 1e derivative V-4. According to this scheme, V-3 is reacted with isobutyl chloroformate in the presence of a suitable amine base, such as triethylamine, diisopropylethylamine, or 4-methylmorpholine,in a neutral solvent, generally CH2Cl2 or THF, to afford an interm~ te mixed anhydride delivalivt;. Without isolation, this derivative is reacted with an a~ pliate pheny!çne~ mine to produce a mono-N-acylated phenylen~ mine inferm/ ~ te. This int~ rm~ t~ is then cyclized to V-4 using acetic acid. Reaction of V-4 with bis(tributyltin) to produce V-5 occurs under palladium catalysis using, for example bis(triphenylphosphine)p~ m (II) chloride ((PPh3)2PdCl2), in an inert solvent, usually DMF. Stille-type coupling of V-2 with V-5 to afford V-6 is m~ ted by a p~ m catalyst, for instance bis(triphenylphosphine)p~ lm (II) chloride ((PPh3)2PdCl2), in the presence of copper (I) iodide (CuI), in a suitable neutral solvent, generally DMF. To obtain V-7, the protecting groups of V-6 are removed by well-established methods known to those of skill in the art and described in the previously cited Greene publication. Thus, the Boc protecting group is removed under acidic conditions, such as 4 M HCl in dioxane or TFA in CH2Cl2, and the methyl ester is hydrolyzed as generally described in Scheme 1 for the conversion of I-7 to I-8.
W O 97/24119 PCT~US96/20748 Scheme VI
CH, HO~, Boc COzCH3 Boc C02CH3 ,CH3 HO ~N
Boc CO2CH3 HO2C ~ = O
N~
Boc CO2CH3 ,CH3 Boc CO2CH3 S a) 3-butyne-1-ol, (PPh3)2PdCl2, PPh3, CuI, Et3N; b) H2, 10% Pd/C, EtOH; c) 2,2,6,6-tetramethyl-oxopiperitlinil-m chloride, CH2Cl2, then NaCl02, Na2HPO37 2-rnethyl-2-butene, H20; d) isobutyl chlc: lvfo~ ate, Et3N, then 1 ,2-phenylene~ mint-" thenAcOH; e) 1.0 N LiOH, THF, H20; f~ TFA, CH2Cl2.
CA 0224l633 l998-06-26 W O 97/24119 PCTrUS96/20748 Compound VI-1, the ~ al~lion of which is described in Scheme V, is reacted with 3-butyne-1-ol in the presence of a catalytic amount of a p~ flinm salt, usually bis(triphenylphosphine)palladium (II) chloride ((PPh3)2PdCl~), together with a catalytic amount of copper (I) iodide (CuI), in an amine solvent, such as S triethylamine (Et3N), to afford VI-2. A phosphine ligand, such as triphenylphosphine (PPh3), can be added to improve the efficiency of the reaction.
Reduction of the acetylenic unit of VI-2 is accomplished under standard hydrogenation conditions which are well-known to those of skill in the art. The rçs-llting compound, VI-3, is oxididized to the corresponding carboxylic acid VI-4, by the two-step method described by Wovkulich (J. Org. Chem. 1993, 58, 832-839).Many alternative m~thofl~ for the oxidation of a primary alcohol to the corresponding carboxylic acid have been described, and can be found in such reference volumes as "Compendium of Organic Synthetic Methods" (published by Wiley-Interscience). Conversion of the carboxylic acid of VI-4 to the ben7imid:~701e derivative VI-5 follows the procedures described in the above Schemes. The methyl ester of VI-5 is removed as described in the above Schemes, and the Boc protecting group is removed under acidic conditions, such as 4 M HClin dioxane or TFA in CH2Cl2, to afford VI-6.
CA 0224l633 l998-06-26 W O 97/24119 PCTrUS96/20748 ~r.hPme VII
HOzC--~ ~ N ~ c ~=~ NH
N ~ ~ N ~ ~
NTs ~ NTs \~/ 3 \~/ 4 N ~ NOH ~ ~_< N ~NOH
~=~ NTs ~ NTs 6 N--O N--O
N ~CO2tBu h N~co2H
NTs ~ NTs ~;~N~CO2Et ~ CO2H
a) l,2-pheny1enP~ mine, DCC, DMF, CH2Cl2; b) AcOH, THF; c) TsCl, NaH, THF;
S d) 03, CH2Cl2, MeOH, then DMS; e) NH20H ~ HCl, NaOAc, MeOH; f) NCS, DMF;
g) tert-butyl 3-butenoate, Et3N; h) 4 M HCI/dioxane, CH2Cl2; i) ethyl 3-aminobutyrate, EDC, HOBt H2O, (i-Pr)2NEt, CH3CN; j) l.0 N LiOH, THF, H2O.
W O 97/24119 PCTrUS96/20748 Commercially available 4-pentenoic acid (VII-l) is converted to the bPn7imi~1~701e derivative VII -2 using to the general procedures described ,~ previously. Protection of one of the nitrogen atoms of the ben7.imid~7ole moiety in VII -2 can be accomplished by reaction with a sulfonyl chloride, for in~t~n~e p-5 toluenesulfonyl chloride, in the presence of a suitable base, generally sodium- hydride or an aqueous alkali metal hydroxide, in an inert solvent, preferably THF, to afford VII -3. ~Itern~tive protecting groups known to those of skill in the art may be used, as long as they are co~ ible with the subsequent chemistry and can be removed when desired. Such protecting groups are described in Greene, "Protective Groups in Organic Synthesis" (published by Wiley-Interscience). Oxidative cleavage of the olefin of VII -3 to afford the aldehyde VII -4 can be conveniently accomplished by ozonolysis in an inert solvent, usually CH2Cl2 or a ll~ Ul'~ of CH2Cl2 and MeOH, followed by in-situ reduction of the ozonide with a suitable reducing agent, generally dimethylsulfide (DMS) or triphenylphosphine. ~ltr.rn~tive mPthocl~ for oxidative cleavage, such as the Lemieux-Johnson reaction (J. Org.
Chem. 1956, 21, 478) can also be used. The aldehyde is converted to the aldoximeVII -5 by standard procedures known to those of skill in the art, and this aldoxime is oxi~li7ed to the oximinoyl chloride derivative VII -6 by the methods described in WO 95tl4682 and WO 95/14683. Reaction of VII -6 with an olefin, such as tert-butyl 3-butenoate (Tet. Lett. 198~;, 26, 381 -384), in the presence of a suitable base, for in.~t~n~e triethylamine or diusopropylethylarnine, in an inert solvent such as benzene or toluene, according to the protocol described in WO 95/14682 and WO
95/14683, gives the cycloadduct I-7. The tert-butyl ester of VII -7 is removed under standard acidic conditions, generally TFA in CH2Cl2 or HCl in dioxane, to give the carboxylic acid VII -8. The carboxylic acid is activated using, for example, EDCand HOBt, or SOCl2, and the activated form is subsequently reacted with an a~prupliate amine, for in.~tAn~e a suitable derivative of ~ nin~, in a neutral solvent, such as DMF, CH2Cl2, or CH3CN, to afford VII -9. Depending on whether acid neutralization is required, an added base, such as diisopropylethylamine ((i-Pr)2NEt) or pyridine, may be used. Many additional methods for converting a W O 97/24119 PCTrUS96/20748 carboxylic acid to an amide are known, and can be found in standard reference books, such as "Compendium of Organic Synthetic Methods", Vol. I - VI (publishedby Wiley-Interscience), or Bodansky, "The Practice of Peptide Synthesis" (published by Springer-Verlag). Derivatives of ,13--alanine are readily available in either racemic 5 or optically pure form by a variety of methods known to those of skill in the art. A
reprçsent~tive method is described in WO 93/07867. The ethyl ester and sulfonyl protecting groups of VII -9 are removed using aqueous base, for example, LiOH inaqueous THF or NaOH in aqueous methanol or ethanol. The interm~ tf .
carboxylate salt is acidified with a suitable acid, for in.ct~nc~.e TFA or HCl, to afford 10 the carboxylic acid VII -10. Alternatively, the int~ te~ carboxylate salt can be isolated, if desired, or a carboxylate salt of the free carboxylic acid can be prepared by methods well-known to those of skill in the art.
Scheme vm I'IH, I ~N ~0 ~CO2Bn ~ ~CO2Bn ~ CO2Bn ,N ~o O CO2Bn CH~f ~C~, ~
~N ~ N ~f ~
S a) COCl2 in toluene, Na2CO3, H20; b) ,B-alanine benzyl ester tosylate, DMAP, - pyridine; c) CH3I, 2,6-lllti~1inç, DMF; d) BrCH2COBr, Et3N, CH2Cl2; e) NaH, DMF, f) CO, (Ph3P)2PdCl2, DIEA, 2-(methylaminomethyl)ben7.imi~1~701e dihydrochloride,NMP; g) H2, Pd/C, EtOH.
W O97/24119 PCTrUS96/20748 Following the procedures of US 5403836 and WO 9504057, except starting from 2-amino-4-iodobenzoic acid (VIII-1) rather than 2-amino-5-iodobenzoic acid,compound VIII -5 is prepared. Reaction of VIII -5 with an ~plopliate amine, for inct~nre 2-(methyl~minom~ thyl)benzimidazole, in a carbon monoxide atmosphere, 5 in the presence of a p~ rlil~m catalyst, preferably (Ph3P)2PdCl2, in an inert solvent, optimally 1-methyl-2-pyrrolidinone (NMP) gives the amide VIII -6. Depending on whether acid neutralization is required, an added base, such as diisopropylethylamine ~DIEA) or pyridine, may be used. The benzyl ester of VIII -6 is removed to afford VIII -7 under standard hydrogenolysis conditions well-known10 to those of skill in the art. ~lt~rn:~tively, the benzyl ester can be saponified using aqueous base, for example, LiOH in aqueous THF, or NaOH in aqueous methanol or ethanol. The intern~ te carboxylate salt is acidified with a suitable acid, for instance TFA or HCl, to afford the carboxylic acid. If desired, the intern~ t~-carboxylate salt of VIII -7 can be isolated, or a suitable salt of the carboxylic acid 15 can be prepared by methods well-known to those of skill in the art.
-6~-Scheme TX
O O
,~~ a ~ ~CO2Et ~~CO2Et ~[~CO2Et ~CO2Et ~ ~CO2Et SMe ¦ N
7 ~
S a) ,B-alanine ethyl ester hydrochloride, DMAP, pyridine; b) BrCH2COBr, Et3N, CH2Cl2; c) NaH, DMF; d) Lawesson's reagent, THF, 50~ C; e) CH3I, (n-Bu)4NHSO4, NaOH, CH2Cll, H20; f~ ~loL)al~ylalI~ine, pyridine HCl, toluene; g) CO, (Ph3P)lPdCl2, DIEA, 2-(methylaminomethyl)benzimidazole dihydrochloride, NMP;
h) LiOH, THF, H20.
-6~ -W O 97124119 PCT~US96/20748 Following the procedures of US 5403836 and WO 9504057, except starting from 4-iodoisatoic anhydride (IX-1, see Scheme I) rather than 5-iodoisatoic anhydride, compound IX-6 is prepared. IX-6 is converted to IX-7 following the procedures described in Scheme XIII for the conversion of XIII -5 to XIII -7.
W O 97/24119 PCTrUS96/20748 SchP,rnP. X
- BocN ~
~, N b 02H l~N~CO2Et HN~
~, N ~
~N~CO2Et ~
H OH H Cl ~N
H N ~ e N ~
N ~ CO2Et ~N
H N
l~, N ~
7 ~ N ~ C02H
., S a) l-Boc-~i~e,azille, NaBH3CN, HCl, MeOH; b) 4 M HCl/dioxane, CH2Cl2; e) SOCl2, CH2Cl2; d) 3, DIEA, DMF; e) 1.0 N NaOH, MeOH.
W O97/24119 PCTrUS96/20748 Reductive amination of the readily available 1-(ethoxycarbonylmethyl)~
piperidone (~-1, EPA 0 542 363 A2) with commercially available l-Boc-pi~e~ le and an ~ p.iate reducing agent, preferably sodium cyanoborohydride, affords amine X-2. The reaction is generally conducted under acidic catalysis, usually with S HCl, in a hydroxylic solvent, for in~tzln~e methanol or ethanol. The Boc protecting group is removed under acidic conditions, preferably HCI/dioxane or TFA, in a suitable solvent, such as CH2Cl2, to the give amine X-3. This reacts with 2-(2-chloroethyl)ben~illlida_ole (X-~;) in the presence of an ~ liate acid scavenger, for instance diisopropylethylamine (DIEA), in a polar solvent, preferably DMF, to give the coupled product X-6. 2-(2-Chloroethyl)bt~.n7;n.icl~7Qle can be preparedfrom 2-(2-hydroxyethyl)b~n7imid~7- 1e by reaction with a suitable halogenating reagent, such as thionyl chloride, or carbon tetrachloride in the presence of triphenylphosphine, in an inert solvent, for example CH2Cl2. The ethyl ester of X-6 is removed using aqueous base, for example, LiOH in aqueous THF or NaOH in 15 aqueous methanol or ethanol. The int~rm~ te~ carboxylate salt is acidified with a suitable acid, for in.stzlnce TFA or HCl, to afford the carboxylic acid X-7.
.~lt~rn~tively, the interTn~ te~ carboxylate salt can be isolated, if desired, or a carboxylate salt of the free carboxylic acid can be prepared by methods well-known to those of skill in the art.
-7~ -Sch~me XI
HN~
~, N ~f ~
~C02tBu OH
N~
NH ~N~
2 CO2tBu OH
--N
NH ~N~
3 ~,CO2H
OH
S a) 2-(3-bromopropyl)bç~7.imi~ 7.ole, DIEA, DMF, b) 4 M HClldioxane, CH2CI2.
The readily available piperazine derivative Xll-l (EPA 0 537 980 Al) is reacted with the readily available 2-(3-bromopropyl)benzimidazole (J. Org. Chem.1962, 27, 2165) in the presence of an a~ ,pliate acid scavenger, for instance 10 diisopropylethylamine (DIEA), in a polar solvent, preferably DMF, to give tnecoupled product XI-2. The tert-butyl ester protecting group is removed under standard acidic conditions, preferably HCl/dioxane or TFA, in a suitable solvent, such as CH2Cl2, to the carboxylic acid XI-3. If desired an a~r~,l)liate salt of the carboxylic acid can be prepared by methods well-known to those of skill in the art.
,.
~ -7~-W O 97/24119 PCTrUS96120748 Sch~.mP.XTT
o ~ N N ~ "" ~ OC 3 b,c ~ ~N~ NH~~O-Bn O O O
NH~N~",~NH ~OH
O O O
S a) 2-(benzirnidazolyl)propionic acid, BOP-Cl, NMM, CH2Cl2; b) LiOH, THF, H20;
c) benzyl ,13-~l~nin~tP, EDC, HOBt H20, NMM, CH2Cl2; d) H2, 10% Pd/C, AcOH, THF, H2O.
The procedures of Beavers et. al., WO 95/25091, Exam~?le 1, were followed to give XII-4, except (2-b~n7imicl~701yl)propionic acid was substituted for Na-Boc-D-lys(Cbz)-OH.
CA 0224l633 l998-06-26 W O97/24119 PCTrUS96/20748 Scheme XIII
~I O
Br ~ CO2CH3 N =< C~2CHs ~3~NH
o ~ C~2H
=~N--W c ~NH
=~ N~ ~ C02CH3 ~NH 4 =~ N~ N ~ CO2H
,~NH 5 a) 2-(arninomethyl)ben7.tmicl:~7.Qle, Et3N, benzene; b) 1.0 N LiOH, MeOH, H20; c) ,13-alanine ethyl ester, BOP, Et3N, CH3CN.
W O 97/24119 PCTrUS96/20748 A suitably functil-n~li7P.d arnine, such as 2-(aminomethyl)ben7imi(l;~7ole~ is reacted with dimethyl 4-bromomethylbenzene-1,3-dicarboxylate (XIII-l;
.y~ esi~ as in EP 0540334Al) under the general conditions described for the ~,c~dLion of l-H-isoindole-5-carb~-x~mitl~7 2,3-dihydro-N-(2-carboxy-ethyl)-2-[2-S (piperidinyl)ethyl]-3-oxo (preparation 1-12, EPA 0 540 334 Al), to afford XIII -2.
The methyl ester of XIII -2 is hydrolyzed using aqueous base, for example, LiOH in aqueous THF or NaOH in aqueous methanol or ethanol, and the int.orm~ t~
carboxylate salt is ~ci~lifi~.-l with a suitable acid, for instance TFA or HCl, to afford the carboxylic acid XIII -3. The carboxylic acid of XIII -3 is converted to an activated form of the carboxylic acid using, for example, EDC and HOBt, SOC12, or BOP reagent, and the activated form is subsequently reacted with an a~ iate amine, for in~t:~nre ,B-alanine ethyl ester, in a suitable solvent such as DMF, CH2Cl2, or CH3CN, to afford XIII -4. Depending on whether acid neutralization is required, an added base, such as diisopropylethylamine ((i-Pr)2NEt) or pyridine, may be used.
Many additional methods for converting a carboxylic acid to an amide are known, ~ and can be found in standard reference books, such as "Compendium of Organic Synthetic Methods", Vol. I - VI (published by Wiley-Interscience), or Bodansky, "The Practice of Peptide Synthesis" ~published by Springer-Verlag). Ester hydrolysis as described above for the conversion of XIII -2 to XIII -3 then affords XIII -S. Alternatively, the int~rm.o~ii~tf~. carboxylate salt of XIII -~ can be isolated, if desired, or a carboxylate salt of the free carboxylic acid can be prepared by methods well-known to those of skill in the art.
CA 0224l633 l998-06-26 PCTrUS96/20748 W O 97/2~119 Scheme XIV
NH~ ~OH N' ~3~OH
'~3'o CO~BnCH~'O CO2Bn d N ~N~
~NH O O
\G/ 5 CO2Bn N ~ ICH~3~
~q~NH O O
\~/ 6 CO2H
a) (Boc)20, NaOH, 1,4-dioxane, HlO; b) BrCH2COlBn, K2C03, acetone; c) 4 M
HCI/dioxane; d) 2-(ben7.imi~1~7.olyl)acetic acid, EDC, DIEA, DMF; e) H2, 5 % Pd/C, MeOH.
XIV-1 is treated with di-tert-butyl dicarbonate and sodium hydroxide in 10 aqueous dioxane to afford XIV -2, which is alkylated on oxygen with benzyl - bromo~f.et~te and potassium carbonate in acetone to give XIV -3. The Boc group in -7~-XIV -3 is removed with hydrogen chloride in dioxane, and the rçsllltin~ XIV -4 is acylated on nitrogen with 29ben7imi-1~7Qlyl)acetic acid, EDC and DIEA in DMF to give XIV -5. The benzyl ester in XIV -5 is cleaved by tre~tml-.nt with hydrogen and Fz~ linm-on-carbon in methanol to give XIV -6.
S
Sch.ome XV
H2N ~ NH ~O
al NH ~ NH~ NH ~f O J3 O O
b NH ~ NH~ NH ~OH
O O
a) 2-(benzirnidazolyl)acetic acid, EDC, DIEA, DMF; b) NaOH, H20, CH30H.
XI-l, prepared as described in Alig et. al., EPA 0372486, is con~ n~e~ with a suitable substituted carboxylic acid, such (2-ben7imitl~701yl)acetic acid, in the 15 presence of EDC and DIEA, and in a suitable solvent, e.g., DMF or acetonitrile.
-7~ -W O97124119 PCT~US96/20748 Many additional methods for converting a carboxylic acid to an amide are known, and can be found in standard reference books, such as "Compendium of Organic ,. Synthesis". Vol. I-VI (published by Springer-Verlag). Hydrolysis of the ester is accomplished by saponirlcation with a suitable reagent, e.g., sodium hydroxide, in a 5 suitable solvent, e.g., aqueous methanol. AAternatively, a benzyl ester may be- converted to the acid by tre~tmfA.nt with a suitable catalyst, e.g., Pd/C, and hydrogen in a suitable solvent, e.g., methanol, ethanol or acetic acid.
Schem~ XVI
1~
~[3~OH
H2N ~ N3 O ~<0 al 1 O-tBu ~3 0 ,<o 2 O-tBu b l ~OH
Q--N o ~W
NH ~W--NH~N~} --COzH
a) 2-(benzill,Adazolyl)acetic acid, EDC, DIEA, DMF; b) TFA
CA 0224l633 l998-06-26 XVI-l, prepared as described in Alig et. al., EPA 0505868, is condensed with a suitable substituted carboxylic acid, such (2-ben7imi~1~7Olyl)acetic acid, in the presence of EDC and DIEA, in a suitable solvent, e.g., DMF or acetonitrile, to give XVI -2. Many additional methods for converting a carboxylic acid to an amide are5 known, and can be found in standard reference books, such as "Compendium of Organic Synthesis". Vol. I-VI (published by Springer-Verlag). Hydrolysis of the ester in XVI -2 is accomplished with trifluoroacetic acid or hydrogen chloride to give XVI -3. Alt~rn~tively, the ester in XVI -2 may be saponified with a suitable reagent, e.g., lN NaOH, in a suitable solvent, e.g., methanol.
Seht-mt- XVII
~IL N N ~ O-tBu H2N 0~O t a l ~0 b l ~ ~ A co H
a) 2-(b~n7imi-1~701yl)acetic acid, EDC, DIEA, DMI;'; b) TFA, CH2Cl2.
W O 97/24119 PCT~US96/20748 XVII-1, prepared as described in Sugihara, et. al., EP 0529858, is condensed with a suitable substituted carboxylic acid, such as (2-bell7imi~1~701yl)acetic acid, to " give XVII -2, and the tert-butyl ester is cleaved with TFA, following the general procedure of Sugihara, et. al., Example 59, to give XVII -3. Many additional - 5 methods for converting a carboxylic acid to an amide are known, and can be found ~ in standard reference books, such as "Compe~-lium of Organic Synthesis", Vol. I-VI
(published by Springer-Verlag).
Schem~ XVITI
Ms-o Q--N
~~~0 a ~ 2 4~ 0 0--t-Bu O-tBu ~0 3 "~0 OH
a) 4-r2-(benzimidazolyl)methyl]phenol, Cs2CO3, DMF; b) TFA
Compound XVIII-1, prepared as described in Himmelsbach, et. al., Australian Patent Application AU-A-86926/91, Example VI~28), is treated with a suitable ~ub~ uled phenol, such as 4-[2-(benzimidazolyl)methyl]phenol, prepared W O 97/24119 PCTrUS96/20748 by the general procedure of Wahlgren and Addison, J. Heterocycl. Chem., 1989, 26, 541-3, following the general method of Himmelsbach, et. al., Example 3(51), to give XVIII -2. The tert-butyl ester in XVIII -2 is hydrolyzed with lN NaOH in CH30H .
following the general procedure of Himmelsbach, Example 7(3) to give XVIII -3.
5 Alternatively, the tert-butyl ester may be cleaved with TFA or HCl.
-8~-W O 97/24119 PCTrUS96/20748 Scheme XIX
N ~CO2CH3 NH2 H ~ H 2 - ,¦, b Q~ N ~ CO2CH3 ~c,d CO2CH3 Q--~ ~~/
'¦, e CO2H
~ ~~
a) HO2CCH2Ph-4-CH2CH~CO2CH3, Ph2POCl, Et3N, DMAP, THF; b) NaH, DMF, BrCH2CO2CH3; c) KOt-Bu, THF, DMF; d) KOt-Bu, CH3I. DMF; e) LiOH, THF, H20.
The procedures of Linz, et. al., EP 0567968, are used to prepare XIX-5, 10 except (2-ben~.imi~1~7.olyl)m~th~n~mine is su~ led for 4-cyanoaniline.
_g~
CA 0224l633 l998-06-26 W O 97/24119 PCT~US96/20748 Sch~me XX
MeO ~C~ MeO ~C~ b NH N ~ CO2Et HO ~ N ~ co2Et R-NHCO ~ e HO2C ~
N ~ CO2Et N ~ CO2Et R-NHCO~C~ R = [~C \~
N ~ CO2Et S a) ClCH2CO2Et, Et3N, DMF; b) BBr3; c) (CF3SO)20; d) CO, Pd(OAc)~, PPh3, DIEA, NMP, NH4HCO3, H20; e) H2N-R, EDC / HOBt, DIEA, DMF; f) H2N-R, CO, Pd(OAc)2, PPh3, DIEA, NMP, NH4HCO3, H20; g) lN NaOH, HOEt Scheme XX provides a method for the preparation of 1,2,3,4-tetrahydroisoquinoline compounds as exemplary fibrinogen receptor antagonists, as described in M. J. Fisher et al. (EP 0635492, Jan. 25, 1995). Accordingly, a 6-methoxy-3,4-dihydroisoquinoline, such as compound XX-~I is prepared by the method described by D. J. Sall and G. L. Grunewald (J. Med. Chem. 1987, 30, - 8~ -.
CA 0224l633 l998-06-26 W O 97/24119 PCT~US96/20748 2208-2216). The isoquinoline is treated with a haloacetic acid ester in the presence of a tertiary amine to afford the 2-acetic acid ester, as exemplified by compound XX-2. The 6-methoxy compound is converted into the corresponding 6-hydroxy compound by methods known in the art, for example with BBr3, which is converted 5 into the triflate with trifluorosulfonic acid anhydride. p~ m catalyzed carbonylation affords the 6-carboxy compound, such as compound XX-5, which is then condensed with an amine, as exemplified by (2-ben7.imi~1~7.olyl)acetic acid, employing a standard amide bond forming reagent to give the desired amide, such as compound XX-6. Saponification affords XX-7. Alternatively, the palladium 10 catalyzed carbonylation reaction with the triflate, exemplified by compound XX-4, may be trapped with said aminomethyl compound to provide, after saponification, the corresponding 6-(2-bçn7.imi~ 7.Qlyl)methylaminocarbonyl compound, XX-7.
Sch~otne XXI
HO ~ CF3SO3~
~N~CO2Et ~N~CO2Et 3 O ~ 4 ~
R-NHCO~ e HO2C~
~N~CO2Et ~N ~CO2Et 6 ~ 5 ~
lg R-NHCO~,~ R = ~N~
N ~ CO2Ft ~J--NH
7 o a) 1. LiN(TMS)2, 2. ClCH2CO2Et, DMF; b) BBr3; c) (CF3SO2)20; d) CO, Pd(OAc)2, PPh3, DIEA, NMP, NH4HC03, H20; e) H2N-R, EDC / HOBt, DIEA, DMF; f) H2N-R
CO, Pd(OAc)2, PPh2, DIEA, NMP, NH4HCO3, H20; g) lN NaOH, HOEt -8~-W O 97/24119 PCT~US96/20748 Scheme XXI provides a method for the pl~al~lion of 3,4-dihydroisoquinolin- l-one compounds as exemplary fibrinogen receptor antagonists, as described M. J. Fisher et al. (~P 0635492, Jan. 25, 1995). Accordingly, the 1-oxo compound XXI-1, prepared by the method described by D. J. Sall and G. L.
Grunewald (J. Med. Chem. 1g87, 30, 2208-2216), is treated with a base, such as LiN(TMS)~, and a haloacetic acid eser to give a 2-acetic acid ester, as exemplified by compound XXI-2. The 1-oxo compound is then employed in the analogous series of reactions deployed in Scheme XX, sub~ g the corresponding 1-oxo analog, as shown in Scheme XXI, to provide XXI-7. As in Scheme XX, alternatively, the p~ m catalyzed carbonylation reaction with the triflate, exemplified by compound XXI-4, may be trapped with an amine to provide, after saponification, the amide, XXI-7.
Sch~mf XXII
H2N~C~ a, b R-CONH~
N ~ CO2tBu N ~ CO2H
~N
~ ~ NH
a) RCO-X; b) TFA / CH2C12 Scheme XXII provides a method for the pr~alaLion of 6-acylaminotetraline compounds as exemplary fibrinogen receptor antagonists, as described M. J. Fisher et al. (;EP 0635492, Jan. 25, 1995). Accordingly, a 6-arnino-2-tert-butyloxyca.1.onyl-tetral-1-one, exemplified by compound XXII-l, which is prepared according to the methods described in M. J. Fisher et al. (EP 0635492, Jan. 25, 1995), is con-l~.n.~ed with an activated derivative of a carboxylic acid, such as the CA 0224l633 l998-06-26 W O 97/24119 PCTrUS96/20748 activated derivative of (2-bPn7imi-~7olyl3acetic acid, to provide, after deesterification, the amide, XXII-2.
Sch~rnP XXlII
HO ~[~G CF3SO3 ~0~ CO2Et R-NHCO~ c HO2C~
COzEt ~ CO2Et 1 e R-NHCO = N~
a) CF3SO2O; b) CO, Pd(OAc)2, PPh3, DIEA, NMP, NH4HCO3, H2O; c) H2N-R, EDC
/ HOBt, DIEA, DMF; d) H2N-R, CO, Pd(OAc)2, PPh3, DIEA, NMP, NH4HCO3, 10 H2O; e) lN NaOH, HOEt Scheme XXIII provides a method for the plc;paldlion of 6-aminoacyltetraline compounds as exemplary fibrinogen receptor antagonists, as described M. J. Fisher et al. (EP 0635492, Jan. 25, 1995). Accordingly, an ethylo~ycall,onylmethyl-6-15 hydroxy-tetral-l-one, exemplified by compound XXIII-l, which is prepared according to the methods described in M. J. Fisher et al. (EP 0635492, Jan. 25, W O 97/24119 PCTrUS96120748 1995), is treated with triffic anhydride to provide the triflate, as exem~ ed bycompound XXIII -2, which is employed in a p~ m catalyzed carbonylation reaction to afford a carboxylic acid, such as compound XXIII -3, which is then condensed with an amine to provide, after deesterifi-~tion, the 6-aminoacyl S compound, XXIII -5. ~AltPrn~tively~ the palladium catalyzed carbonylation reaction with the triflate exemplified by compound XXIII -2, may be trapped with said amine compound to provide, after saponification, the corresponding 6-aminoacyl compound, XXII~ -5.
Scheme XXIV
02N~ EtO IP~o S CO2Et 4 ,/
e H2N ~ + ~ CO2Et 1 f~9 1 f'9 R-CONH~ RCONH~ ~ CO2H
.~ N \
a) BrCH2CO2Et, KzCO3~ NaI; b) 1. DBU, EtOH, 2. HCl, EtOH; c) DiBAL, -78 ~ C; d) 15 NaH, THF; e)Hz, 10% Pd-C; f) R2CO-X; g)lN NaOH, MeOH
-8~-W O97/24119 PCTrUS96120748 Scheme XXIV provides a method for the preparation of 5-acylaminobenzofuran and S-acylaminodihydrobenzofuran compounds as exemplary r fibrinogen receptor antagonists, as described in M. L. Denney, et al. (~P 0655439, 31/5/95). Accordingly, a 4-nitrosalicylaldehyde, exemplified by compound XXIV-1 5 , is treated with a haloacetic acid ester to give the phenoxyacetic acid ester, exemplified by compound XXIV -2. A 2-alkoxycarbonylfuran, exemplified by compound XXIV -3, is obtained by treating the aldehyde with base, for example with DBU. The 2-alkoxycarbonyl group is reduced to the aldehyde, for example with DiBAB. Wittig reaction affords the 2-acrylate ester, exemplified by compound 10 XXIV -4, which is reduced to the benzofuran-2-propionic acid ester, exemplified by compound XXIV -5 and the dihydrobenzofuran-2-propionic acid ester, exemplified by co~ )o-llld XXIV -6. The amine XXIV -~ is then condensed with an activated derivative of a carboxylic acid to provide, after deesterification, the amide 5, XXIV -8. ~lt~rnzltively, the amine XXIV -6 is condensed with an activated derivative of a 15 carboxylic acid to provide, after dççstçrifiç~tion, the amide, XXIV -7.
-8~-W O97/24119 PCTrUS96/20748 Schem~ XXVa HO ~ CO2Et ~ CO2Et b,c a-l a-2 TBDMSO ~ CHO TBDMSO ~
EtO2P ~ a-~ ~ CO2Et ~e,f HO ~ CO2Et ~ CO2Et a-6 a-7 a)l.TBDMS-Cl, imid~ole; b)DiBAl-H, -78 ~C, d) NaH, THF; e) H2, 5% Pd-C; f) Et4N+ F -CA 0224l633 l998-06-26 W O 97/24119 PCTrUS96/20748 Scheme XXVb HO ~ CF3SO3 ~ CO2Et a-6 b-1 d/
~' I b R-NHCO~ CO~EtCO2Et b-3 b-2 l e R-NHCO~co2H R- [~NH
b-4 a) ~CF3SO2)20; b) CO,Pd(OAc)2, PPh3, DTF.A, NMP, NH4HCO3, H20; c) H2N-R, EDC / HOBt, DIEA, DMF; d) H2N-R, CO, Pd(OAc)2, PPh2, DIEA, NMP, NH4HCO3, H20 e) lN NaOH, EtOH
W O 97/24119 PCT~US96/20748 SchemP XXVc CO2Et a-7 c-1 ~ 1 R-NHCO ~ CO2Et CO2Et l e R-NHCO~co2H R =[~NH
a) (CF3SO2)20; b) CO, Pd(OAc)2, PPh3, DlF.A, NMP, NH4HC03, H20; c) H~N-R, EDC / HOBt, DIEA, DMF; d) H2N-R, CO, Pd(OAc)2, PPh2, DIEA, NMP, NH4HCO3, H20 e) lN NaOH, BtOH
Scheme XXV provides a method for the ~rt;p~dtion of 5-aminoacylbenzofuran and 5-aminoacyldihydrobenzofuran compounds as exemplary fibrinogen receptor antagonists, as described in M. L. Denney, et al. (EP 0655439, 31/5/95). Accordingly, a 5-hyd~ ybellzofuran-2-carboxylic acid ester, such as compound XXVn-l, prepared in the manner of M. L. Denney, et al. (~P 0655439, 31/5/95), is treated with TBDMS-Cl to provide the TBDMS delivalive of the ester,XXVa-2. The ester is reduced to an aldehyde, such as co,llpolJ.ld XXVa-3. Wittig 9~
W O 97/24119 PCT~US96/2074.~.
reaction affords the acrylic acid ester, XXVa-4. Catalytic reduction affords thebenzofuran-2-acetic acid ester and the dihydl. bell of uran-2-acetic acid ester.r Cleavage of the silyl ether group of each ester, by methods known to the art, affords either the benzofuran-2-acetic acid ester, XXVa-~, or the dihydrobenzofuran-2-S acetic acid ester, XXVa-6.
~ As seen in Scheme XXVb and XXVc, each alcohol in turn may be converted to a carboxylic acid via p~ m catalyzed carbonylation, such as compound XXVb-2 or XXVc-2, which is then condensed with an amine to provide, after ~lees~.; rication, the amide XXVb-4 or XXVc-4. Alternatively, the p~ lm 10 catalyzed carbonylation reaction with the triflate exemplified by compound XXVb-1, or XXVc-1, may be trapped with said aminomethyl compound to provide, after deesterification, the corresponding 6-aminoacyl compound, XXVb-4 or XXVc-4.
Amide coupling reagents as used herein denote reagents which may be used 15 to form peptide bonds. Typical coupling methods employ carbo-liimicles, activated anhydrides and esters and acyl halides. Reagents such as EDC, DCC, DPPA, PPA, BOP reagent, HOBt, N-hydroxysuccinimi(le and oxalyl chloride are typical.
Coupling methods to form peptide bonds are generally well known to the art.
The methods of peptide synthesis generally set forth by Bodansky et al., THE
20 PRACTICE OF ~ SYNTHESIS, Springer-Verlag, Berlin, 1984, Ali et al. in J. Med. Chem., 29, 984 (1986) and J. Med. Chem., 30, 2291 (1987) are generally illustrative of the technique and are incorporated herein by reference.
Typically, the amine or aniline is coupled via its free arnino group to an a~L.,~,iate carboxylic acid substrate using a suitable carbodiimide coupling agent, 25 such as N,N' dicyclohexyl carbodiimide (DCC), optionally in the presence of catalysts such as l-hydroxybenzotriazole (HOBt) and dimethylamino pyridine (DMAP). Other methods, such as the formation of activated esters, anhydrides or acid h~licles, of the free carboxyl of a suitably protected acid substrate, and subsequent reaction with the free amine of a suitably protected amine, optionally in 30 the presence of a base, are also suitable. For example, a protected Boc-amino acid or _9~
W O 97/24119 PCTrUS96/20748 Cbz-amidino benzoic acid is treated in an anhydrous solvent, such as methylene chloride or tetrahydrofuran(THF), in the presence of a base, such as N-methyl morpholine, DMAP or a trialkylamine, with isobutyl chlc,loroll,late to form the "activated anhydride", which is subsequently reacted with the free amine of a second 5 protected amino acid or aniline.
The compounds of formula (XIX) and (XX) are commercially available or are prepared by methods known in the art such as illustrated herein disclosed insta~ldard reference books, like the COMPENDIUM OF ORGANIC S~rHETIC METHODS, Vol. I-VI (Wiley-Interscience). A generally applicable route to ben7imifl~7oles is disclosed in Nestor et al, J. Med. Chem. 1984, 27, 320. Representative methods for preparing compounds of formula (XX) are also common to the art and may be found, for instance, in EP-A 0 381 033.
Acid addition salts of the compounds are prepared in a standard manner in a suitable solvent from the parent compound and an excess of an acid, such as 15 hydrochloric, hydl~lvll~ic, hydrofluoric, sulfuric, phosphoric, acetic, trifluoroacetic, maleic, succinic or methanesulfonic. Certain of the compounds form inner salts or zwitterions which may be acceptable. Cationic salts are prepared by treating theparent compound with an excess of an ~lk~lint? reagent, such as a hydroxide, carbonate or alkoxide, containing the ~lupliate cation; or with an appl.,p~iate 20 organic amine. Cations such as Li+, Na+, K+, Ca++, Mg++ and NH4+ are specific examples of cations present in ph~rms~reutir~lly acceptable salts.
- This invention also provides a pharm~-~entical composition which comprises a compound according to formula (I)-(V) and (XXI)-(XXII) and a ph~rm~reutically acceptable carrier. Accordingly, the compounds of formula (I)-(V) and (XXI)-25 (XXII) may be used in the m~nuf~tllre of a m~-lic~m~nt. Pharmaceutical compositions of the compounds of formula (I)-(V) and (XXI)-(XXII) prepared as hereinbefore described may be form~ te~l as solutions or lyophilized powders forparenteral ~rlmini~tration~ Powders may be recon.ctitnte~1 by addition of a suitable diluent or other ph~rm~entic~lly acceptable carrier prior to use. The liquid 30 fi~rm~ tion may be a buffered, isotonic, aqueous solution. Examples of suitable diluents are normal isotonic saline solution, standard 5% dextrose in water or _ g W O 97/24119 PCTrUS96t20748 buffered sodium or ammonium acetate solution. Such formulation is especially suitable for parenteral ~rlmini~tration, but may also be used for oral ~rlmini~tration or cont~inP~l in a metered dose inhaler or nebulizer for in~llffl~tion It may be desirable to add excipients such as polyvinylpyrrolidone, gelatin, hydroxy cellulose, acacia, polyethylene glycol, m~nnitol, sodium chloride or sodium citrate.
Alternately, these compounds may be encapsulated, tableted or prepared in a emulsion or syrup for oral ~lmini~tration Ph~rm~eeutiç~lly acceptable solid or liquid carriers may be added to enh~nre or stabilize the composition, or to facilitate plG~ald~ion of the composition. Solid carriers include starch, lactose, calcium sulfate dihydrate, terra alba, m~gn~ m stearate or stearic acid, talc, pectin, acacia, agar or gelatin. Liquid carriers include syrup, peanut oil, olive oil, saline and water.
The carrier may also include a sllct~inP~l release m~teri:~l such as glyceryl monostearate or glyceryl distearate, alone or with a wax. The amount of solid carrier varies but, preferably, will be between about 20 mg to about 1 g per dosage unit. The pharmaceutical pl~l,aldLions are made following the conventional techniques of pharmacy involving milling, mixin~, gr:~n~ ting, and compressing, when necessary, for tablet forms; or milling, mixing and filling for hard gelatin capsule forms. When a liquid carrier is used, the ~lepaldlion will be in the form of a syrup, elixir, emulsion or an aqueous or non-aqueous suspension. Such a liquid fnrmnl~tion may be ~flmini.~tered directly p.o. or filled into a soft gelatin capsule.
For rectal ~lmini~tration, the compounds of this invention may also be combined with excipients such as cocoa butter, glycerin, gelatin or polyethyleneglycols and molded into a suppository.
The compounds described herein are antagonists of the vitronectin receptor, and are useful for treating diseases whe~ the underlying pathology is attributable to ligand or cell which interacts with the viLIoile~;lin receptor. For instance, these compounds are useful for the treatment of diseases wherein loss of the bone matrix creates pathology. Thus, the instant compounds are useful for the tre~tment of ostoeporosis, hyperparathyroidism, Paget's disease, hypercalcemia of mz~ n~3ncy,- 30 osteolytic lesions produced by bone m~t~t~ , bone loss due to immobilization or sex hormone deficiency. The compounds of this invention are also believed to have 9~
W O 97/24119 PCTrUS96/20748 utility as ~ntitl~mor, anti-angiogenic, ~ntiinfl~mm~tory and anti-mPt~t~tic agents, and be useful in the treatment of atherosclerosis and restenosis.
The compound is ~lmini~tered either orally or L)a-ellLe,dlly to the patient, in a manner such that the concentration of drug is sufficient to inhibit bone resorption, or S other such indication. The ph:lrm~re~lti(~l composition cont~ining the peptide is ~lmini~tered at an oral dose of between about 0.1 to about 50 mg/kg in a manner conci~tent with the condition of the patient. Preferably the oral dose would be about 0.5 to about 20 mg/kg. For acute therapy, parenteral ~r~ . alion is prefeilcd. An intravenous infusion of the peptide in 5% dextrose in water or normal saline, or a 10 similar formulation with suitable excipients, is most effective, although an cc~ r bolus injection is also useful. Typically, the parenteral dose will be about 0.01 to about 100 mg/kg; preferably between 0.1 and 20 mg/l~g. The compounds are ~lmini~tered one to four times daily at a level to achieve a total daily dose of about 0.4 to about 400 mg/kg/day. The precise level and method by which 15 the compounds are ~lmini.~tf red is readily fletermin~d by one routinely skilled in the art by comparing the blood level of the agent to the concentration required to have a therapeutic effect.
The compounds may be tested in one of several biological assays to determine the concentration of compound which is required to have a given 20 I)h~ ological effect.
Tnhi'~it JI~ of vilrv~ binding Solid-Phase [3H7-sK&F-107260 Binding to av~3: Human placenta or human platelet av~B3 (0.1-0.3 mg/mL) in buffer T (cont~ining 2 mM CaC12 and I %
octylglucoside) was diluted with buffer T collt;1i.. i.. g 1 mM CaC12, 1 mM MnC12, 1 mM MgC12 (buffer A) and 0.05% NaN3, and then imm~ t~ly added to 96-well ELISA plates (Corning, New York, NY) at 0.1 rnL per weIl. 0.1 - 0.2 ~g of ocv,133 was added per well. The plates were incubated overnight at 4~C. At the time of the experiment, the wells were washed once with buffer A and were incubated with 0.1 mL of 3.5% bovine serum alburnin in the same buffer for 1 hr at room W O 97/24119 PCT~US96/20748 telllpeldLuie. Following in~nb:~tion the wells were aspirated completely and washed twice with 0.2 mL buffer A.
Compounds were dissolved in 100% DMSO to give a 2 mM stock solution, which was diluted with binding buffer (15 mM Tris-HCl (pH 7.4), 100 mM NaCl, 1 mM CaC12, 1 mM MnC12, 1 mM MgC12) to a final compound concentration of 100,uM. This solution is then diluted to the required final compound concentration.
Various concentrations of unlabeled antagonists (0.001 - 100 ~M) were added to the wells in triplicates, followed by the addition of 5.0 nM of [3H]-SK&F-107260 (65 -86 Ci/mmol).
The plates were incubated for 1 hr at room lelllpeldLul~. Following incubation the wells were aspirated completely and washed once with 0.2 mL of ice cold buffer A in a well-to-well fashion. The receptors were solubilized with 0.1 mL
of 1% SDS and the bound [3H]-SK&F-107260 was determin~-1 by liquid scin~ tion counting with the addition of 3 mL Ready Safe in a Beckman LS Liquid Scintill~tic~n Counter, with 40% efficiency. Nonspecific binding of [3H3-SK~F-107260 was ~et~rmined in the presence of 2 ,uM SK&F-107260 and was consistently less than 1 % of total radioligand input. The ICso (concentration of the antagonist to inhibit 50% binding of [3H]-SK&F-107260) was determined by a nonlinear, least squares curve-fitting routine, which was modified from the LUNDON-2 program.
The Ki (dissociation constant of the antagonist) was calculated according to theequation: Ki = ICso/( l + L/Kd), where L and Kd were the concentration and the dissociation constant of [3H]-SK&F-107260, respectively.
Compounds of the present invention inhibit vitronectin binding to SK&F
107260 in the concentration range of about 0.001 to 50 micromolar.
Compounds of this invention are also tested for in vitro and in vivo bone resorption in assays standard in the art for ev~ ting inhibition of bone formation, such as the pit formation assay disclosed in EP 528 587, which may also be performed using human osteoclasts in place of rat osteoclasts, and the ovarectomi7ed - rat model. described by Wronski et al., Cells and Materials 1991, Sup. 1, 69-74.
_95 W O 97124119 PCT~US96/20748 Vascular smooth linuscle cell ~ ..lion assay Rat or human aortic smooth muscle cells were used. The cell migration was monitored in a Transwell cell culture chamber by using a polycarbonate membrane with pores of 8 um (Costar). The lower surface of the filter was coated with vitronectin. Cells were suspended in DMEM supplemented with 0.2% bovine serum albumin at a concentration of 2.5 - 5.0 x 106 cells/mL, and were pretreated with test compound at various concentrations for 20 min at 20~C. The solvent alone was used as control. 0.2 mL of the cell suspension was placed in the upper c~ nlent of the chamber. The lower co~ lllent contained 0.6 rnL of DMEM supplemented with 0.2% bovine serum albumin. lncubation was carried out at 37~C in an atmosphere of 95% air/5% CO2 for 24 hr. After incubation, the non-migrated cellson the upper surface of the f1lter were removed by gentle scraping. The filter was then fixed in methanol and stained with 10% Giemsa stain. Migration was measuredeither by a) counting the number of cells that had rnigrated to the lower surface of the filter or by b) extracting the stained cells with 10% acetic acid followed by ~terrninin~ the absorbance at 600 nM.
PARATHYROIDE~TOMIZED RAT MODEL
Each experimental group consists of 5-6 male Sprague-Dawley rats. The rats are parathyroidectomized (by the vendor, Taconic Farms) 7 days prior to use. Twenty four hours prior to use, circ~ tin~ ionized calcium levels are measured in whole blood immediately after it has been withdrawn by tail venipuncture into hep~rini7f r1 tubes. Rats are included if ionized Ca level (measured with a Ciba-Corning model 634 calcium pH
analyzer) is _1.2 mM/L. The rats are then put on a diet of calcium-free chow anddeionized water. At the start of the experiment the rats weigh approximately 100g.
Baseline Ca levels are measured and the rats are ~irnini~tered control vehicle (saline) or compound (dissolved in saline) as a single intravenous (tail vein) bolus injection followed r im mf.~ t~ly by a single ~ubc~lt:~nPous injection of either human paldlhyl~id hormone 1-34 peptide (hPI'Hl-34, dose 0.2mg/kg in saline/0.1% bovine serum albumen, Bachem, Ca) _9~_ W O97/24119 PCT~US~6120748 or the PTH vehicle. The calcemic response to PTH (and any effect of compound on this response) is measured 2h after co~ oulld/PTH allmini~tration.
.
RAT ULNA DRII~T MODEL
Each experimental group consists of 8-10 male Sprague-Dawley or Wistar rats of aL~plo~illlately 30-40g body weight at the start of the e~elilllent. The agent being tested is ~rlmini~trred by an a~prop.iate route as single or multiple daily doses for a period of seven days. Prior to ~lmini~tration of the first dose, the rats are given a single dose of a 10 fluorescent marker (tetracycline 25mg/lcg, or calcein lOmg/lcg) that labels the position of bone forming surfaces at that point in time. After dosing of compound has been completed, the rats are killed and both forelimbs are removed at the elbow, the foot is removed at the ankle and the skin removed. The sample is frozen and mounted vertically on a microtome chuck. Cross sections of the mi(l~h~ft region of the ulna are cut in the 15 cryostat. The rate of bone resorption is measured morphom~-tric~lly in the medial-dorsal portion of the cortical bone. The mea~ule.llellt is done as follows: the amount of bone resorbed at the periosteal surface is equal to the t1ict~nre by which the periosteal surface has advanced towards the fluorescent label which had been incorporated at the endosteal bone formation surface on day zero; this fli~t~nre is calculated by subtracting the width of 20 bone between the label and the periosteal surface on day 7 from the width on day zero; the resorption rate in microns per day is calculated by dividing the result by 7.
HUMAN OSTEOCLAST RESORPTION ASSAY ("PIT ASSAY") 25 ~ Aliquots of osteoclastoma-derived cell suspensions are removed from liquid nitrogen strorage, warmed rapidly at 37~C and washed x l in RPMI- 1640 m~ m by centrifugation (lOOOrpm, 5 mins at 4~C).
~ Aspirate the medium and replace it with murine anti-HLA-DR antibody, diluted 1:3 in RPMI-1640 ml rlillm Incubate for 30 mins on ice and mix the cell suspension .~
frequently.
_ W O 97/24119 PCTrUS96/20748 ~ The cells are washed x2 with cold RPMI-1640 by c~ L irugation (1000 rpm, 5 rnins at 4~C) and the cells are transferred to a sterile 15 rnl centrifuge tube. The number of mononuclear cells are enumerated in an il~l~roved Nellh~ r counting chamber.
S
~ Sufficient m~n~tic beads (5 / mononuclear cell), coated with goat anti-mouse IgG, are removed from their stock bottle and placed into 5 ml of fresh mP~ lm (this washes away the toxic azide preservative). The m~ lm is removed by immobilizing the beads on a magnet and is replaced with fresh medium.
~ The beads are mixed with the cells and the suspension is incub~t~cl for 30 mins on ice. The suspension is mixed frequently.
~ The bead-coated cells are immobilized on a magnet and the rem~ining cells 15 (osteoclast-rich fraction) are dec~nt~d into a sterile 50 ml centrifuge tube.
~ Fresh mto-lillm is added to the bead-coated cells to dislodge any trapped osteoclasts.
This wash process is repeated xlO. The bead-coated cells are discarded.
20 ~ The osteoclast~s are enullleldt~d in a counting chamber, using a large-bore disposable plastic pasteur to charge the chamber with the sample.
~ The cells are pelleted by centrifugation and the density of osteoclasts ~dj-lste~l to 1.5xlO4/ml in EMEM m~ lm, supplemented with 10% fetal calf serum and 25 1.7gllitre of sodium bicarbonate.
~ 3ml aliquots of the cell suspension ( per tre~tmP~t) are ~ieC~ntl ~i into 15mlcenkifuge tubes. The cells are pelleted by centrifugation.
30 ~ To each tube 3m1 of the a~p~ .;ate treatment are added (diluted to 50 uM in the EMEM medium). Also included are ~pl~,pliate vehicle controls, a posiLive conkol gg W O 97/24119 ~T~US96/20748 (87MEM1 diluted to 100 ug/ml) and an isotype control (IgG2a diluted to 100 ug/ml). Incubate at 37~C for 30 mins.
~, ~ 0.5ml aliquots of the cells are seeded onto sterile dentine slices in a 48-well plate - 5 and incubated at 37~C for 2 hours. Each treatment is screened in quadruplicate.
., ~ The slices are washed in six changes of warm PBS (lO ml / well in a 6-well plate) and then placed into fresh tre:~tmf~nt or control. Incubate at 37~C for 48 hours.
10 Tartrate l~- ~;C~nt acid Ph~3s~ ce (TRAP) Pl~ce-lul~ (selective stain for cells of the osteoclast lineage).
~ The slices are washed in phosphate buffered saline and fixed in 2% gluteraldehyde (in 0.2M sodium cacodylate) for 5 mins.
~ They are washed in water and incubated in TRAP buffer for 5 mins at 37~C.
~ Following a wash in cold water they are incubated in cold acetate buffer / fast red garnet for 5 mins at 4~C.
~ Excess buffer is aspirated, and the slices are air dried following a wash in water.
~ The TRAP positive osteoclasts are enumerated by bright-field microscopy and are then removed from the surface of the dentine by sonication.
~ Pit volumes are dele~ ed using the Nikon/Lasertec ILM2 1 W confocal microscope.
Human o~leo~lr--l le3~ ~io.. and adhesion assavs 30 Pit resorption and adhesion assays have been developed and standardized usingnormal human osteoclasts derived from osteoclastoma tissue. The osteoclast _99_ W O 97/24119 PCT~US96/20748 population is negatively selected from the osteoclastoma cell suspensions using magnetic beads (Dynal Inc, NY). These beads are coated with a murine monoclonal antibody that recognizes a hurnan class II major histocompatihility antigen that is present on a large number of mononuclear cells in the cell suspensions. The cells 5 that express this antigen, and consequently bind the beads, are removed from the mixture of cells using a m~n~t The osteoclast-rich suspension is then ready to use in the assays det~ below.
Resor~lio.- sssay (with ELISA readout) 10 Enriched preparations of osteoclasts are preincubated for 30 minllfes at 37~C with test compound (4 doses) or controls. They are then seeded onto bovine cortical bone slices in wells of a 48-well tissue culture plate and are in~ubat-~-l for a further 2 hours at 37nC. The bone slices are washed in six changes of warm phosphate buffered saline (PBS), to remove non-adherent cells, and are then returned to wells of a 48 15 well plate con~ining fresh compound or controls. The tissue culture plate is then incubated for 48 hours at 37"C. The supern~t~n~c from each well are aspirated into individual tubes and are screened in a cornpetitive ELISA that detects a collagen peptide that is released during the resorption process. This is a commercially available ELISA (Osteometer, Denmark) that contains a rabbit antibody that 20 specifically reacts with an 8-amino acid se~uence (Glu-Lys-Ala-His- Asp-Gly-Gly-Arg) that is present in the carboxy-tçrmin~l telopeptide of the al-chain of type I
collagen. The results are expressed as % inhibition of resorption colllpal~;d to a vehicle control.
25 Adhesion assay Osteoclastoma-derived osteoclasts are preincubated with compound (4 doses) or controls at 37~C for 30 minutes. The cells are then seeded onto osteopontin-coated slides (human or rat osteopontin, 2.5ug/ml) and incubated for 2 hours at 37~C. Non adherent cells are removed by washing the slides vigorously in P~S and the cells30 r~m~ining on the slides are fixed in acetone. The osteoclasts are stained for tartrate-resistant acid phosphatase (TRAP), a selective marker for cells of this phenotype, and are enumerated by light microscopy. The results are expressed as % inhibition of adhesion colll~alcd to a vehicle control.
-10~
.
W O 97/24119 PCTrUS96120748 Inhibition of RGD-m~ t.-d GPIIb-IIIa binding pllrific~tion of GPIIb-ma Ten units of ollt~l~tP-l, washed human platelets (obtained from Red Cross) were lyzed by gentle stirring in 3% octylglucoside, 20 mM Tris-HCl, pH 7.4, 140 mM NaCl, 2 mM CaC12 at 4~C for 2 h. The lysate was centrifuged at lOO,OOOg for 1 h. The sUpprn~t~nt obtained was applied to a 5 mL lentil lectin sepharose 4B
column (E.Y. Labs~ preequilibrated with 20 mM Tris-HCl, pH 7.4, 100 mM NaCl, 2 mM CaC12, 1 % octylglucoside (buffer A). After 2 h incubation, the column was washed with 50 rnL cold buffer A. The lectin-retained GPIIb-IIIa was eluted withbuffer A containing 10% dextrose. All procedures were performed at 4~C. The GPIIb-IIIa obtained was >95% pure as shown by SDS polyacrylamide gel electrophoresis .
Incorporation of GPIIb-IIIa in Liposomes.
A mixture of phosphatidylserine (70%) and phosphatidylcholine (30%) (Avanti Polar Lipids) were dried to the walls of a glass tube under a stream of nitrogen. Purified GPIIb-IIIa was diluted to a final concentration of 0.5 mg/rnL and mixed with the phospholipids in a protein:phospholipid ratio of 1:3 (w:w). The mixture was resuspended and sonicated in a bath sonicator for 5 min. The mixturewas then dialyzed overnight using 12,000- 14,000 molecular weight cutoff dialysis tubing against a 1000-fold excess of 50 mM Tris-HCl, pH 7.4, 100 mM NaCl, 2 mM
CaC12 (with 2 changes). The GPIIb-IIIa-cont~ining liposomes wee centrifuged at 12,000g for 1~ min and resuspended in the dialysis buffer at a final protein concentration of approximately 1 mg/mL. The liposomes were stored at -70C until needed.
Competitive Binding to GPIIb-IIIa The binding to the fibrinogen receptor (GPIIb-ma) was assayed by an indirect competitive binding method using [3H]-SK&F-107260 as an RGD-type ligand. The binding assay was performed in a 96-well filtration plate assembly (Millipore Corporation, Bedford, MA) using 0.22 um hydrophilic durapore membranes. The wells were precoated with 0.2 mL of 10 ,ug/mL polylysine (Sigma Chemical Co., St. Louis, MO.) at room temperature for 1 h to block nonspecific 35 binding. Various concentrations of unlabeled ben7~ 7~rines were added to the -1~1-CA 0224l633 l998-06-26 W O 97/24119 PCT~US96/20748 wells in quadruplicate. [3H]-SK&F-IQ7260 was applied to each well at a final concentration of 4.5 nM, followed by the addition of 1,ug of the purified platelet GPIIb-IIIa-cont~ining liposomes. The mi~ulcs were incubated for 1 h at room tel,,pcldture. The GPIIb-ma-bound [3H]-SK&F-107260 was seperated from the S unbound by filtration using a Millipore filtration manifold, followed by washing with ice-cold buffer (2 times, each 0.2 rnL). Bound radioactivity rem:~ining on the filters was counted in 1.5 mL Ready Solve (Beckm~n Instr lm~nt~, Fullerton, CA) in a Beckman Liquid Scintillation Counter (Model LS6800), with 40% efficiency.
Nonspecific binding was ~et~ in.o~t in the presence of 2 ,uM unlabeled SK&F-107260 and was con~i~te-ntly less than 0.14% of the total radioactivity added to the samples. All data points are the mean of quadruplicate ~leterrnin~tions.
Competition binding data were analyzed by a nonlinear least-squares curve fitting procedure. This method provides the IC50 of the antagonists (concentration of the antagonist which inhibits specific binding of [3H]-SK&F-107260 by 50% at equilibrium). The IC50 is related to the equilibrium dissociation constant (Ki) of the antagonist based on the Cheng and Prusoff equation: Ki =
IC50/(l~L/Kd), where L is the concentration of t3H~-SK&F-107260 used in the co~llpeLiLive binding assay (4.5 nM), and Kd is the dissociation constant of r3H]-SK&P-107260 which is 4.5 nM as ~ rmin~l by Scatchard analysis Compounds of the present invention inhibit the vitronectin binding to SK&P
007260 with a Ki at the vitronectin receptor that is about ten-fold greater than that for the fibrinogen receptor. Preferred compounds have a Ki at the vitronectin receptor that is thirty-fold greater than that at the fibrinogen receptor. The most preferred compounds have a Ki at the vitronectin receptor that is a hundred-foldgreater than that at the fibrinogen receptor.
The examples which follow are intended in no way to limit the scope of this invention, but are provided to illustrate how to make and use the compounds of this invention. Many other embodiments will be readily a~cnt to those skilled in the art.
W O 97/2411g PCTrUS96/20748 G~o,n~ral Nuclear magnetîc resonance spectra were recorded at either 250 or 400 MHz using, respectively, a Bruker AM 250 or Bruker AC 400 spectrometer. CDC13 is deuteriochloroform, DMSO-d6 is hPxAfle~ riodimethylsulfoxide, and CD30D is tetradeuteriomethanol. Chemical shifts are reported in parts per million (o) downfield from the internal standard tetramethylsilane. Abbreviations for NMR
data are as follows: s=singlet, d=doublet, t=triplet, q=quartet, m=multiplet, dd=doublet of doublets, dt=doublet of triplets, ~ arent, br-broad. J inclic~tes the NMR coupling constant measured in Hertz. Continuous wave infrared (IR) spectra were recorded on a Perkin-Elmer 683 infrared spectrometer, and Fourier transform infrared (FTIR) spectra were recorded on a Nicolet Impact 400 D infrared spectrometer. IR and FTIR spectra were recorded in trAn~mi~ion mode, and band positions are reported in inverse wavenumbers (cm-l). Mass spectra were taken oneither VG 70 FE, PE Syx API III, or VG ZAB HF instruments, using fast atom bombardment (FAB) or ele~ v~L~ldy (ES) ionization techniques. Elemental analyseswere obtained using a Perkin-Elmer 240C elemental analyzer. Melting points were taken on a Thomas-Hoover melting point apparatus and are uncorrected. All te~ ela~ules are reported in degrees Celsius.
~nAltt~ch Silica Gel GF and E. Merck Silica Gel 60 F-254 thin layer plates were used for thin layer chromatography. Both flash and gravity chromatography were carried out on E. Me~ck Kieselgel 60 (230-400 mesh) silica gel. Analytical and preparative HPLC were carried out on Rainin or Berkm~n chromatographs. ODS
refers to an octadecylsilyl derivatized silica gel chromatographic support. 5 ,u Apex-ODS in~lic~ s an octadecylsilyl derivatized silica gel chromatographic support having a nominal particle size of 5 ,u, made by Jones Chromatography, Littleton,Colorado. YMC ODS-AQ~ is an ODS chromatographic support and is a registered tra(1~mArk of YMC Co. Ltd., Kyoto, Japan. PRP-l~) is a polymeric (styrene-divinylbenzene) chromatographic support, and is a registered trademark of T~milton Co., Reno, Nevada) Celite(~) is a filter aid composed of acid-washed diatomaceous silica, and is a registered trAtl~m~rk of Manville Corp., Denver, Colorado.
Methyl (+)-7-carboxy4-methyl-3-oxo-2,3,4,5-tetrahydro- lH- 1,4-- 35 benzodi~epine-2-acetate, methyl (2S)-7-carboxy-4-methyl-3-oxo-2,3,4,5-tetrahydro- 1 H- 1,4-benzodiazepine-2-acetate, methyl (2R)-7-carboxy-~methyl-3-oxo-2,3,4,5-tetrahydro-lH-1,4-ben7o~ 7~pine-2-acetate, methyl (+)-7-carboxy-4-isopropyl-3-oxo-2,3,4,5-tetrahydro- 1 H- 1,4-benzodiazepine-2-acetate, methyl (+~-7-carboxy-3-oxo-2-(2-phenylethyl)-2,3,4,5-tetrahydro- 1 H- 1,4-benzodiazepine-2-acetate, methyl (+)-8-carboxy-2-methyl-3-oxo-2,3,4,5-tetrahydro-lH-2-benzazepine-4-acetate, methyl (+) 7-amino-5-oxo-~(2-phenylethyl)-lH-1,4-5 benzodiazepine-2-acetic acid, and tert-butyl 4-fluoro-3-methylbenzoate were prepared by the method of Bondinell, et al., WO 93/00095. Methyl (~)-7-carboxy-4-(2-methoxyethyl)-3-oxo-2,3,4,5-tetrahydro- 1 H- 1,4-benzodiazepine-2-acetate, methyl (~)-7-carboxy-4-[2-(3,4-methylenedioxyphenyl)ethyl] -3-oxo-2,3,4,5-tetrahydro- 1 H-1,4-benzodiazepine-2-acet~tf- m~thyl (+)-7-carboxy-3-oxo-2,3,4,5-tetrahydro-1~I-1,4-benzodiazepine-2-acetate, methyl (+)-2,3,4,5-tetrahydro-7-[[[(ben7imi~l~7Ql-2-yl)methyl~amino]carbonyl]-4-methyl-3-oxo-lH-1,4-benzodiazepine-2-acetate, (S)-2,3,4,5-tetrahydro-7-t[[(bf~n7:imi-1~7.ol-2-yl)methyl]methylamino]carbonyl]~
methyl-3-oxo-lH-1,4-benzodiazepine-2-acetic acid, 2-(methylaminomethyl)ben~imic~7ole dihydrochloride, and 4-aza-5-rnethyl-2-(methylamino)methylben7imif~70le were prepared according to P50256-1.
Preparation 1 Preparation of 2-(aminomethyl)-4-~a-5-methylben7.imi-1~7--1e dihydrochloride a) 2,3-Diamino-6-methylpyridine 10% Pd/C (3.2 g, 3 mmole) was added to a solution of 2-Amino-6-methyl-3-nitropyridine (2.30 g, 15 mmole) in absolute EtOH (150 mL), and the mixture was shaken at RT under H2 (50 psi). After 1.5 hr, the mixture was filtered through celite~), and the filtrate was concentrated under vacuum to afford the title compound as a yellow oil. This was used without further purification: 'H NMR (250 MHz, CD30D) ~ 6.82 (d, lH), 6.36 (d, IH), 2.25 (s, 3H).
b) 2-Amino-3-[(benzyloxycarbonyl)glycyl~amino-6-nlethylL)ylidine DCC (3.09 g, 15 mmole) was added to a solution of 2,3-diamino-6-methylpyridine (15 mmole) and Cbz-glycine (3.14 g, 15 mmole) in DMF (19 mL) and CH2Cl2 (19 mL) at 0~C under argon. When the DCC had dissolved, the slightly cloudy solution was warrned to RT. After 18.5 hr, the rnixture was filtered through celite(~, and the filtrate was concentrated to dryness on the rotavap. The residue was reconcentrated from xylenes (to remove DM~) to leave a yellow solid. Silica gel chromatography (10% MeOH/CHCl3) gave the title compound (2.24 g, 48%) as a yellow solid: TLC Rf (10% MeOH/CHCl3) 0.57; 'H NMR (250 MHz, DMSO - d6) o W O 97/24119 PCTrUS96/20748 9.11 (br s, 1 H), 7.48 - 7.60 (br t, 1 H), 7.20 - 7.48 (m, 6 H), 6.40 (d, 1 H), 5.69 (br s, 2 H), 5.06 (s, 2 H), 3.82 (d, 2 H), 2.23 (s, 3 H).
c) 4-Aza-2-(benzyloxycarbonyl)aminomethyl-5-methylbenzimidazole S A solution of 2-amino-3-[(benzyloxycarbonyl)glycyl]amino-6-melhyl~ylidine (2.24 g, 7.13 mmole) in glacial AcOH (70 mL) was heated to refluxunder argon. After 17 hr, the solution was concentrated (rotavap, high vacuum), and the residue was reconcentrated from toluene (to remove AcOH). The r~slllting yellow oil was treated with hot EtOAc (20 mL) and the ll~i~lUlG was cooled to RT.
The solid was collected by suction filtration and washed with EeOAc to afford the title compound (1.72 g, 81%) as an off-white solid: TLC R, (15% MeOH/CHCl3) 0.63; MS (ES) m/e 297.4 (M + H)+.
d) 2-(Arninomethyl)-4-aza-5-methylben7imi-l~7ole dihydrochloride 10% Pd/C (153 mg, 0.14 mmole) was added to a solution of 4-aza-2-(benzyloxycarbonyl)aminomethyl-5-methylben7imi(1~7Ole (213.4 mg, 0.72 mmole) and 1.0 N HCl (1.44 mL, 1.44 mmole) in absolute EtOH (7.2 mL). The llli~lult; was purged with H2, then was stirred briskly at RT under H2 (balloon). After 2 hr, the reaction was filtered through celite, and the filtrate was concentrated on the rotavap to leave the title compound as an off-white solid: MS (ES) m/e 163.2 (M + H)+.
rr~l)aidlion 2 Preparation of methyl (+)-2.3~4.5-tetrahydro-7-carboxy-4-~3.3-dimethylbutyl)-3-oxo- lH- 1.4-ben7.odiazepine-2-acetate a) tert-Butyl 3-[(3,3-dimethylbutyl)amino]methyl-4-nitrobenzoate A ~ u~e of tert-butyl 3-methyl-4-nitrobenzoate (WO 93/00095; 17.7 g, 74.7 mmol), NBS (19.9 g, 112.0 mmol), benzoyl peroxide (1.81 g, 7.47 mmol), and CC14 (370 mL) was heated at reflux. After 17.5 h, the reaction was cooled thoroughly in ice and filtered to remove the pleci~ilaled succinimicle. The filtrate was concentrated to leave a brownish-yellow oil.
This oil ~4.2 g, 13.29 mmol), was dissolved in dry THF (50 mL), and 3,3-dimethylbutylamine (3.0 g, 29.64 mmol) was added all at once. The orangish-yellow solution was stirred at RT for 80 min, then was concentrated to remove the THF. The residue was diluted with Et20 (150 mL) and washed sequentially with 1.0 N NaOH (25 mL) and H2O (25 mL). The combined aqueous layers were back-W O 97/24119 PCT~US96120748 extracted with Et2O (50 mL~, and the combined organic layers were washed with brine (25 mL) and dried (MgS04). Concentration gave the crude title compound (4.19 g, 94%~ as a light-brown oil: MS (ES) rn/e 337.2 (M+H)+.
S b) tert-Butyl 3-[[N-(3,3-dimethylbutyl)-N-(tert-butoxycarbonyl)]amino]methyl-4-nitrobenzoate Di-tert-butyl dicarbonate (4.0 g, 18.39 mmol) was added all at once to a solution of tert-butyl 3-[(3,3-dimethylbutyl)amino]methyl-4-nitrobenzoate (4.12 g, 12.26 mmol) in CHCl3 (80 mL) at RT. After 18 h, the reaction was concen~lated and reconcentrated from hexanes (to remove CHCl3). Silica gel chromatography (10-25% EtOAc/hexanes) gave the title compound (5.0 g, 93%) as a yellow oil: MS
(ES) m/e 437.2 (M+H)+, 459.2 (M+Na)+.
c) tert-Butyl 4-amino-3-[~N-(3,3-dimethylbutyl)-N-(tert-butoxycarbonyl)]amino]methyl benzoate 10% Pd/C (1.0 g, 0.94 mmol) was added to a solution of tert-butyl 3-[[N-(3,3-dimethylbutyl)-N-(tert-butoxycarbonyl)]amino]methyl~-nitrobenzoate (4.95 g,11.35 mmol) in EtOAc (50 mL), and the mixture was shaken on a Parr apparatus at RT under H2 (55 psi). After 4 h, the reaction was filtered through celite~, and the filtrate was concentrated to afford the title compound (4.3 g, 93%) as a reddish-brown oil: MS (ES) m/e 407.4 (M+H)+.
d) tert-Butyl (~t)-4-[2-(1,4-dimethoxy-1,4-dioxobutyl)amino~-3-[[N-(3,3-dimethylbutyl)-N-(tert-butoxycarbonyl)]amino]methylbenzoate A solution of tert-butyl 4-amino-3-[[N-(3,3-dimethylbutyl)-N-(tert-butoxycarbonyl)]arnino]methylbenzoate (5.6 g, 13.79 mmol) and dimethylacetylene dicarboxylate (1.86 mL, 15.17 mmol) in MeOH (28 mL) was heated at reflux for 1 h, then was cooled to RT. The rçslllting solution was combined with MeOH (80 mL) and 10% Pd/C (2.9 g, 2.76 mmol), and the mixture was shaken on a Parr apparatus at RT under H2 (50 psi). After 22 h, the reaction was filtered throughcelite~), and the filtrate was concentrated on the rotavap. The residue was reconcentrated from CHC13 (to remove MeOH), then was chromatographed on silica gel (25% EtOAc/hexanes). The title compound (2.64 g, 42%) was obtained as a faintly yellow oil: MS (ES) m/e 551.2 (M+H)+.
e) Methyl (+)-2,3,4,5-tetrahydro-7-carboxy-4-(3,3-dimethylbutyl)-3-oxo-lH-1,4-benzodiazepine-2-acetic acid PCTrUS96120748 TFA (25 mL) was added all at once to a solution of tert-butyl (+)4-~2-(1 ,4-~iimPth~xy-1 ,4-dioxobutyl)amino]-3-~[N-(3,3-dimethylbutyl)-N-(tert-butoxycarbonyl)]amino]methylbenzoate (12.64 g, 4.8 mmol) in anhydrous CH2Cl2 (25 mL) at 0~C, and the faintly yellow solution was warmed to RT. After 1 h, thesolution was concenLlaLed on the rotavap, and the residue was reconcentrated from toluene (to remove residual TFA). The resulting oil was combined with toluene (50 mL) and Et3N (3.34 mL, 24 mmol), and the mixture was heated to reflux. A light yellow, homogeneous solution was produced. After 16 h, the reaction was concentrated on the rotavap to leave a solid residue. This was dissolved in a minimllm of MeOH (ca. 10 mL), diluted with H2O (10 mL), and acidif1ed with glacial AcOH to pH 4.5. The ~ lulc was filtered, and the precipitate was washed sequentially with MeOH and E~t20, then was dried in high vacuum to afford the title compound (1.88 g, 93%) as a nearly colorless powder: MS (ES) m/e 363.2 (M+H)+.
~lGI)aldlion 3 Preparation of Bisr(ben7imi-1~7ol-2-yl)methyllamine tris(trifluoroacetate) a) Bis[[1-N-(tert-butoxycarbonyl)bçn7imi(1~7Ol-2-yl]methyl]-N-(tert-butoxycarbonyi)amine To a stirred solution of 2-aminomethylbenzimidazole dihydrochloride hydrate (6.26 g, 28.4 mmol) and triethylamine (4.0 mL, 28.4 mmol) in dry THF (50mL) was added l-(tert-butoxycarbonyl)-2-(bromomethyl)bçn7imit1~7c 1e (P50256-1;
2.00 g, 9.48 mmol) in THF (30 mL),. After 8 h, a solution of di-tert-butyl dicarbonate (10.0 g, 45.84 mmol~ in CHC13 (50 mL) was added slowly. The resulting mixture was stirred at RT overnight then was concentrated. The residuewas taken up in CH2C12 (150 mT.), and washed sequentially with water (60 rnL), 5%
NaHCO3 (60 mL), and brine (60 mL). Drying (MgSO4), concentration, and silica gel chromatography (6% MeOH/CH2C12) gave the title compound (0.46 g, 8%) as a faint yellow oil: MS (ES) m/e 578.4 (M+H)+.
b) Bis[(bPn7imic1:~701-2-yl)methyl]amine tris(trifluoroacetate) A solution of TFA (3 mL) and CH2C12 (9 mL) at RT was added all at once to bis[[1 -N-(tert-butoxycarbonyl)benzimidazol-2-yl]methyl}-N-(tert-butoxycarbonyl)amine (0.23 g, 0.4 mmol). After 35 min, the solution was concentrated on the rotavap, and the residue was reconcentrated from toluene (to 107 W O 97/24119 PCTrUS96~0748 remove residual TFA) to afford the title compound (0.17 g, 68%) as an off-white powder: MS (ES) m~e 278.0 ~M+H)+.
Preparation 4 Preparation of 2-rrl-r(benzimidazol-2-yl)methyllben7imi(1~7ole~methyllamine bi~(Jri lluorQacetate) a) [[l-N-(tert-Butoxycarbonyl)be~7imi-1~701-2-yl3methyl]-N-(tert-butoxycarbonyl)amine To a stirred solution of 2-aminomethylben7imidzl701e dihydrochloride hydrate (3.0 g, 13.63 mmol) and triethylamine (8.44 rnL, 61.3 mmol) in dry CH~C12 (50 mL) a solution of di-tert-butyl dicarbonate (6.54 g, 30.0 mmol) in CH2Cl2 (50 rnL) was added at 0~C. The reaction was stirred at RT for 1 h, then more of triethylamine (1.9 mL, 13.8 mmol) and di-tert-butyI dicarbonate (2.97 g, 13.63 mmol) were added. The reslllting mixture was stirred at RT for 24 h, then was concenkated. The residue was taken up in CH2Clz (50 mL) and washed sequentially with 0.5 N HCl (2x40 mL), 5% NaHC03 (50 mL), and brine (50 mL). The crude product was recrys~lli7ecl from CH2Cl~/ether to give the title compound (2.8 g, 59%) as a wnite powder: MS (ES) m/e 348.2 (M~H)+.
b) 2-[[1-[[( 1-(tert-Butoxycarbonyl)ben7imirl~701-2-yl3methyl]ben7.imitl~7.01e]methyl]-N,N-di-(teLt-butoxycarbonyl)amine To a stirred solution of [[l-N-(tert-butoxycarbonyl)bPn7imirl~7Ql-2-yl]methyl]-N-(di-tert-butoxycarbonyl)amine (0.6 g, 1.73 mmol) and NaH (0.1 g, 4.17 mmol) in dry THF (12 mL) and DMF (4 mL) was added 1-(tert-butoxycarbonyl)-2-(bromomethyl)ben7imi~ 701e ~0.6 g, 1.93 nimol). Theresulting Lul~ was stirred at RT for 1 h, then was concenLIdL~d. The residue was taken up in CH2C12 (100 mL) and washed sequentially with water (50 mL), 5% NaHC03 (30 mT ), and brine (30 mL). Drying (MgS04), concentration, and silica gel chromatography (2:3 EtOAc/hexanes) gave the title compound (0.27 g, 27%) as a faint yellow oil: MS (ES) m/e 578.2 (M~H)+.
c) 2-~[l-[(Ben7imi~1~7-)l-2-yl)methyl]bçn7imitl~7~1e~methyl]amine bis(trifluoro~et~1r) A solution of TFA/CH2C12 (30 ml, 25%) at RT was added all at once to 2-[[1-[[( 1 -N-tert-butoxycarbonyl)bel-7 i ~ 701-2-yl]methyl]ben7imi~1~701e]methyl]-N,N-CA 0224l633 l998-06-26 W O 97/24119 PCT~US96/20748 di-(tert-butoxycarbonyl)amine (0.25 g, 0.43 mmol). After 25 min, the solution was concentrated on the rotavap, and the crude product was recryst~ d from CH2Cll/ether to give the title co~ oulld (0.17 g, 63%) as an off-white powder:
MS (ES) m/e 278.0 ~M+H)+.
~xamI)le 1 a~dLion of (+)-2.3~4.5-tetrahydro-7-rrr(4-~a-5-methylbellzi.ll~dazol-2-yl)methyllaminolcarbonyll -4-(2-methoxyethyl)-3-oxo- 1 H- 1.4-benzodiazepine-2-a~tic acid a) Methyl (+)-2,3,4,5-tetrahydro-7-[[[(4-aza-5-methylben7imiclzl7Ql-2-yl)methyl]amino]carbonyl]~(2-methoxyethyl)-3 -oxo- 1 H- 1,4-benzodiazepine-2-acetate EDC (138 mg, 0.72 mrnole) was added to a solution of methyl (+)-7-carboxy-4-(2-methoxyethyl)-3-oxo-2,3,4,5-tetrahydro- lH- 1,4-benzodiazepine-2-acetate (202 mg, 0.60 mmole), 2-(arninomethyl)-4-aza-5-methylben7imicl~7Ole dihydrochloride (0.72 mmole), HOBt ~ H2O (97 mg, 0.72 mmole), and diisopropylethylamine (0.84 mL, 4.8 mmole) in anhydrous CH3CN (3 mL) at RT.
After 16 hr, the reaction was concentrated, and the residue was reconcellllaL~d from xylenes/CHCl3. Silica gel chromatography (15% MeOH/CHCI3) gave the title compound (impure): TLC Rf (15% MeOH/CHCI3) 0.55; MS (ES) m/e 481.5 (M +
H)t. This was used without further purification.
b) (+)-2,3,4,5-Tetrahydro-7-[tt(4-aza-S-methylben7imi~1~7ol-2-yl)methyl]amino~carbonyll-4-(2-methoxyethyl)-3-oxo- 1 H- 1,4-benzodiazepine-2-acetic acid A two-phase mixture of methyl ( )-2,3,4,5-tetrahydro-7-[[[(4-aza-5-methylbel.~.i...ic1sl7.Ql-2-yl)methyl]amino]carbonyl]-4-(2-methoxyethyl)-3-oxo-lH-1,4-benzodiazepine-2-acetate (0.60 mrnole), 1.0 N LiOH (1.8 mL, 1.8 mmole), and THF (4.2 mL) was stirred at RT for 45 min, then was concentrated to remove the THF. The aqueous layer was washed with Et2O (2 x 2 mL), and the Et2O layers werediscarded. The aqueous layer was diluted with CH3CN (2 mL) and acidified with TFA (0.23 mL). The res-llting solution was concentrated to dryness on the rotavap, and the residue was purified by ODS chromatography (12~o CH3CN/H20 contS~ini 0.1% TFA (250 mL), then 15% CH3CN/H20 cont:~ining 0.1% TFA). Concentration and lyophilization gave the title compound (199.5 mg, 50% for two steps) as a light W O 97/24119 PCT~US96/20748 yellow powder: HPLC (PRP~ , 15% CH3CN/H20 cont~ining 0.1%
TFA) K' = 1.4; MS (ES) rn/e 467 (M + H)'. Anal. Calcd for C23El26N6Os ~ 1.5 CF3COzH 1.33 H2O: C, 47.21; H, 4.60; N, 12.70. Found: C, 47.20; H, 4.73; N, 12.79.
S .
Examl~le 2 Pl~a~tion of (_)-2~3~4~s-tetr~ydro-7-rrr(ben7im~ 7~ol-2-yl)methyllaminolcarbonyll-4-(2-methoxyetl~ -3-oxo-lH-1.4-ben7.o-lJa7epine-2-acetic acid a) Methyl (_)-2,3,4,5-tetrahydro-7-[[t(bel ,7i . l . if 1~7f~l-2-yl)methyl]arnino]carbonyl]
4-(2-methoxyethyl)-3-oxo- lH- 1 ,4-benzodiazepine-2-acetate EDC (230 mg, 1.2 mmole) was added to a solution of methyl (+)-7-carboxy-4-(2-methoxyethyl)-3-oxo-2,3,4,5-tetrahydro- lH- 1 ,4-benzodiazepine-2-acetate (336.4 mg, 1.0 mmole), 2-(aminomethyl)be~7imif1~7f-1e dihydrochloride hydrate (264 mg, 1.2 mmole), HOBt ~ H2O (162 mg, 1.2 mmole), and diisopropylethylamine (0.70 mL, 4.0 mmole) in anhydrous DMF (5 mL) at RT. After 17 hr, the reaction was concentrated, and the residue was reconcentrated from xylenes (2 x) to remove DMF. The residue was diluted with H2O (3 mL) and extracted with CHCl3 (3 x 5 mL). The combined extracts were treated with MeOH (2 rnL) to dissolve a precipitate, then were dried (MgSO,) and concentrated. Reconcentration from xylenes (to remove residual DMF) left a light yellow solid. This was dissolved in MeOH/CHCI3, and the solution was concentrated to leave an oil. Silica gel chromatography (10% MeOH/CHCl3) gave an off-white solid, which was triturated with EtOAc (3 mL) to afford the title compound (397.1 mg, 85%) as a colorless solid: TLC R, (10% MeOH/CHCl3) 0.46; MS (ES) m/e 466.2 (M + H)~.
b) (_)-2,3,4,5-Tetrahydro-7-[~[(ben7imi(1~7Ol-2-yl)methyl]amino]carbonyl]-4-(2-methoxyethyl)-3-oxo- lH- 1 ,4-benzodiazepine-2-acetic acid 1.0 N LiOH ( 1.0 mL, 1.0 mmole) was added to a suspension of methyl (+)-2,3,4,5-tetrahydro-7-[[[~benzimidazol-2-yl)methyl3amino]carbonyl]~(2-methoxyethyl)-3-oxo-lH-1,4-ben7o~ 7f-pine-2-acetate (397 mg, 0.85 mmole) in THF (4.3 rnL) and H20 (3.3 mr ) at RT. The light yellow mixture was stirred at 40 -50~C for 1 hr, and the reslllting homogeneous solution was then stirred at RT for 17.5 hr. The reaction was concentrated, and the rçslllting oil was dissolved in H20 (4 mL). The solution was filtered to remove particnl~tes, and the filtrate was CA 0224l633 l998-06-26 W O97/24119 PCTrUS96/20748 neutralized with 1.0 N HCl (1.0 rnL). The yellowish solid was collected and triturated with good stirring with hot 1: 1 CH3CN/H2O. The resulting solid was collected, washed with plenty of 1: 1 CH3CN/H2O, and dried in high vacuum (40~C)to afford the title compound (327.9 mg, 85%) as a colorless powder: HPLC (PRP-S l(E~), 15% CH3CN/H20 cont:~inin~ 0.1% TFA) K' = 4.6; MS (ES) m/e 452.2 (M +
H)~. Anah Calcd for C23H25N5O5: C, 61.19; H, 5.58; N, 15.51. Found: C, 61.18; H,5.58; N, 15.39.
~ExamE)le 3 Preparation of (+)-4-~4-~rr(lH-~en7~imicla7Ql-2-yl)methyllmethylaminolcarbonyllphenyll-3-phenylbutanoic acid a) Ethyl 3-hydroxy-4-(4-methoxyphenyl)-3-phenylbutanoate Anhydrous EtOAc (4.3 mL, 44 mmole) was added dropwise over 5 - 6 rnin to a solution of lithium bis(trimethylsilyl)amide (1.0 M in THF, 40 mL, 40 mmole) in dry THF (60 mL) in a flame-dried flask at -78~C under argon. The yellow solution was stirred at -78~C for 0.5 hr, then a solution of 2-(4-methoxyphenyl)-1-phenyleth~n<)nP (Chem. Ber. 195S, 91, 755-759; 4.53 g, 20 mmole) in dry THF (20 ml) was added dropwise over 12 rnin. Additional THF (2 mL) was used in transfer.After 0.5 hr, The reaction was quenched with satd NH4Cl (120 mL) and warmed to RT. EtOAc extraction, drying (MgSO4), concentration, and silica gel chromatography (20% EtOAc/hexanes) gave the title compound (6.13 g, 96%) as a light yellow oil: TLC Rf (20% EtOAc/hexanes) 0.34; MS (ES) m/e 315.2 (M + H)~.
b) Ethyl 4-(4-methoxyphenyl)-3-phenylbutanoate Boron trifluoride etherate (4.8 mL, 39 mmole) was added dropwise over 3 rnin to a solution of ethyl 3-hydroxy-4-(4-methoxyphenyl)-3-phenylbutanoate (6.13 g, 19.5 mmole) and triethylsilane (6.2 mT, 39 mmole) in anhydrous CH2C12 (49 rnL) at 0~C under argon. The reaction was stirred at RT overnight, then was quenched with 5% NaHCO3 (100 mL). The ll~i~lul~ was stirred briskly for 10 min, then was separated. The aqueous layer was extracted with CH2Cl2 (100 mL), and the combined organic layers were dried (Na2SO4) and concentrated. The residue was reconcentrated from hexanes (to remove CH2Cl2) to leave a yellow oil. This was - 35 dissolved in absolute EtOH (100 mT.), and 10% Pd/C (775 mg, 1.95 mmole) was added. The mixture was shaken on a Parr apparatus at RT under H2 (50 psi) for 2 hr, then was filtered through celite~3). The filtrate was concentrated, and the residue was W O 97/24119 PCT~US96/20748 chromatographed on silica gel (15 % EtOAc/hf~x~n~s). The title compound (5.27 g, 91%) was obtained as a colorless oil: TLC Rr (15% EtOAc/hexanes) 0.40;
MS (ES) m/e 299.2 (M + H)+.
c) Ethyl 4-(4-hydroxyphenyl)-3-phenylbutanoate Anhydrous ~ minllm trichloride (4.49 g, 33.7 mmole) was added all at once to solution of ethyl 4-(4-methoxyphenyl)-3-phenylbutanoate (2.01 g, 6.74 mmole) and eth~nethiol (2.5 mT., 33.7 mrnole) in anhydrous CH2Cl2 (67 mL) at 0~C under argon. The yellow solution was warmed to RT and stirred for 3 hr, then was recooled to 0~C and ~luen~hl cl with cold 3 N HCl (67 mL). The mixture was stirred for 5 rnin, then was separated. The aqueous layer was extracted with CH2Cl2 (2 x100 mL), and the combined organic layers were dried (Na2SO4) and concentrated.
Silica gel chromatography (25% EtOAc/hexanes) gave the title compound (1.84 g, 96%) as a colorless oil: TLC Rf (30% EtOAc/hexanes) 0.47; MS (ES) m/e 285.2 (M
+ H)'.
d) Ethyl 3-phenyl-4-[4-(trifluoromethanesulfonyloxy)phenyl]butanoate Trifluorom~th~n.oslllfonic anhydride (1.4 mL, 8.4 mrnole) was added rapidly -dropwise to a solution of ethyl 4-(4-hydroxyphenyl)-3-phenylbutanoate (1.84 g, 6.47 rnmole) and 2,6-lutidine (1.5 mL, 12.9 mmole) in anhydrous CH2Cl2 (32 mL) at -78~C under argon. After 0.5 hr, the yellow solution was warmed to RT and stirredfor 1 hr. The reaction was diluted with Et2O (150 mL) and washed sequentially with 1.0 N HCl (15 mL), 5% NaHCO3 (15 mL), and saturated brine(15 mL). Drying (MgS04), concentration, and silica gel chromatography (15% EtOAc/hexanes) gave the title compound (2.62 g, 97%) as a nearly colorless oil: TLC Rf (20%
EtOAc/hexanes) 0.55; MS (ES) m/e 417.0 (M + H)~.
e) Ethyl 4-(4-carboxyphenyl)-3-phenylbllt:~n~-~t~
A mixture of ethyl 3-phenyl~-[4-~trifluornm~th:-nesulfonyloxy)phenyl]butanoate (2.62 g, 6.29 mmole), anhydrous KOAc (2.47 g, 25.16 mmole), Pd(OAc)2 (70.6 mg, 0.31 mrnole), dppf (697.4 mg, 1.26 mmole), and anhydrous DMSO (31 mL) was purged with carbon monoxide (three evacuation/ carbon monoxide purge cycles, followed by bubbling carbon monoxide through the mixture for 5 min), then was heated at 70~C under a balloonof carbon monoxide. After 3.5 hr, the reaction was diluted with H2O (31 mL), cooled in ice, and acidified with 1.0 N HCl (25 mL). CH2C12 extraction (2 x 100 mL), drying (MgSO4), concentration, and reconcentration from toluene left a :
W O 97/24119 PCT~US96/20748 reddish-orange liquid. Silica gel chromatography (1% AcOH in 7:3 toluene/EtOAc) gave the title compound (1.78 g, 91 %) as a cream-colored solid:
TLC Rf (1% AcOH in 7:3 toluene/EtOAc) 0.47; MS (ES) m/e 313.2 (M + H)+.
S f) Ethyl (+)-4-[4-[~[(lH-bell~hllidazol-2-yl)methyl]methylamino~carbonyllphenyl]-3 -phenylbutanoate - EDC (230 mg, 1.2 mmole) was added to a solution of ethyl 4-(4-carboxyphenyl)-3-phenylbutanoate (312.4 mg, 1.0 rnrnole), 2-(methylaminomethyl)ben7imi(1z-7ole~dihydrochloride (281 mg, 1.2 mmole), HOBt lQ H2O (162 mg, 1.2 rnmole), and diisopropylethylamine (0.70 mL, 4.0 mmole) in anhydrous CH3CN (5 mL) at RT. After 18 hr, the reaction was concentrated, and the brown residue was chromatographed on silica gel (5% MeOH in 1: 1 EtOAc/CHCl3).
The title compound (439.2 mg, 96%) was obtained as a light orange foam: TLC Rf (5% MeOH in 1: 1 EtOAc/CHCl3) 0.50; MS (ES) m/e 456.2 (M + H)+.
g) (~)~[4-[[[(}H-Benzimidazol-2-yl)methyl]methylamino]carbonyl]phenyl]-3-phenylbutanoic acid A solution of ethyl (_)-4-[4-[[[(lH-ben7imitl~7ol-2-yl)methyl]methylamino]carbonyl]phenyl~-3-phenylbutanoate (439.2 mg, 0.96 20 mmole) and 1.0 N NaOH (1.2 mL, 1.2 mmole) in EtOH (8.4 mL) was stirred at 50~C. After 24 hr, the reaction was concentrated to dryness and the residue was purified by ODS chromatography (35% MeOH/H20). Concentration and lyophilization gave the title compound (412.2 mg, 86%) as a colorless powder:
HPLC (PRP-l(~), 35% CH3CN/H20 cont~inin~ 0.1% TFA) K' = 1.4; MS (ES) 428 25 (M + H)', 450 (M + Na)~. Anal. Calcd for C26H2JN3O3Na 2.75 H2O: C, 62.58; H, 5.96; N, 8.42. Found: C, 62.34; H, 5.84; N, 8.44.
Rx~m,l?le 4 30 ~le~)a dlion of (+~-4-14-rrr(ben7imi~1~7~-1-2-yl)methyllmethylaminolcarbonyllphenyll-3-(dimethylaminocarbonyl)butanoic acid a) tert-Butyl 4-bromobenzoate Trifluoromethanesulfonic acid (0.18 rnL, 2 rnmole) was added dropwise to a - 35 mixture of 4-bromobenzoic acid (20.10 g, 100 rnrnole), anhydrous CH2Cl2 (100 mL), and condensed isobutylene (-78~C; 100 mL), and the rf s~ in~ mixture was allowedto reflux under a dry ice/acetone condenser. After 40 min, more isobutylene (30 CA 0224l633 l998-06-26 W O 97/24119 rcTrusg6/2o748 mL) was added, and reflux was continued for an additional 20 min. The reaction was poured into Et2O (500 mL) and washed sequentially with 1.0 N KOH (2x 50 rnL), H2O (50 mL), and satd brine (50 mL). Drying (MgSO4), concentration, and silica gel chromatography (5% EtOAc/hexanes) gave the title compound (15.28 g, 59%) as a light yellow oil: TLC Rf (5% EtOAc/hexanes) 0.59; MS (ES) m/e 259/257 (M + H)+.
b) Methyl 3-[4-(tert-buto~y~;all~onyl)phenyl]propenoate A solution of tert-butyl 4-hromobenzoate (5.14 g, 20 mmole), methyl - 10 acrylate (9.1 rnL, 100 mrnole), Pd(OAc)2 (224.5 mg, 1 rnrnole), tri-o-tolylphosphine (608.8 mg, 2 rnmole), and diisopropylethylamine (7.0 mL, 40 mmole) in propionitrile (100 rnL) was heated at reflux for 3 hr, then was concentrated on the rotavap. The residue was diluted with Et2O (200 mL) and washed sequentially with1.0 N HCl (2 x 50 mT), 5% NaHCO3 (50 rnL), and satd brine (50 mL). Drying (MgSO4), concentration, and silica gel chromatography (15% EtOAc/hexanes) gave the title compound (3.34 g, 64%) as a light yellow solid: TLC Rf (20%
EtOAc/hexanes) 0.51; MS (ES) rn/e 263.0 (M + H)+.
c) Methyl 3-[4-(tert-butoxycarbonyl)phenyl]propanoate 10% Pd/C (2.71 g, 2.55 mrnole) was added to a solution of methyl 3-[4-(tert-butoxycarbonyl)phenyl]propenoate (3.34 g, 12.73 mmole) in EtOAc (65 rnL) and MeOH (65 rnL), and the mixture was shaken on a Parr a~paldLus at RT under H2 (50psi). After 3 hr, the reaction was filtered through celite~, and the rlltrate was concentrated to dryness on the rotavap. Reconcentration from hexanes left the title compound (3.27 g, 97%) as a cloudy, grayish oil: TLC Rf (20% EtOAc/hexanes) 0.63; MS (ES) m/e 265.0 (M + H)+.
d) 3-t4-(tert-Butoxycarbonyl)phenyl]propanoic acid A rnixture of methyl 3-[4-(tert-butoxycarbonyl)phenyl]propanoate (3.27 g, 12.37 rnrnole), 1.0 N LiOH (14.8 mL, 14.8 mmole), THF (31 mL), and H2O (16 mL) was stirred at RT for 1.5 hr, then was concentrated on the rotavap to remove theTHF. The aqueous solution was washed with Et20 ~2 x 30 rnL), and the Et,O layerswere discarded. The aqueous layer was acidified with 1.0 N HCl (ca. 17 mL), and the mixture was extracted with CHCl3 (3 x 50 mL). Drying (Na2SO4) and concent;ation gave the title compound (3.04 g, 98%) as a colorless powder: mp 88.5 - 89.5~C; MS (DCVNH3) m/e 268.0 (M ~ NH4)+.
W O 97/24119 PCT~US96/20748 e) N,N-Dimethyl 3-[4-(tert-butoxycarbonyl)phenyl]prop~n~mi~l EDC (2.09 g, 10.88 mmole) was added to a solution of 3-[4-(tert-butoxycarbonyl)phenyl]propanoic acid (2.27 g, 9.07 mmole), di~llelllylamine hydrochloride (0.88 g, 10.88 mmole), HOBt ~ H2O (1.47 g, 10.88 mmole), and diisopropylethylamine (3.2 rnL, 18.14 mmole) in anhydrous CH3CN (45 mL) at RT.
After 19.5 hr, the reaction was concentrated, and the residue was chromatographed on silica gel (EtOAc). The title compound (2.46 g, 98%) was obtained as a colorless oil: TLC Rf (EtOAc) 0.52; MS (ES) mte 278.4 (M + H)~.
f) Ethyl 4-[4-(tert-butoxycarbonyl)phenyl]-3-(dimethylaminocarbonyl)butanoate A solution of lithium bis(trimethylsilyl)amide in THF (1.0 M, 5.8 mL, 5.8 mmole) was added dropwise over 2.5 min to a solution of N,N-dimethyl 3-[4-(tert-butoxycarbonyl)phenyl]prop~n~mi~lf~ (1.34 g, 4.83 mmole) in dry THF (48 mL) at -78~C under argon. The yellow solution was stirred at -78~C for 0.5 hr, then ethyl bromoacetate (2.7 mL, 24.15 mmole) was added over 15 sec down the walls of the flask (to precool). After 0.5 hr, the reaction was poured into satd NH4Cl (50 rnL), and the mixture was extracted with EtOAc (2 x 100 mL). Drying (MgSO4), concentration, and reconcentration from xylenes left a light yellow oil. Silica gel chromatography (1: 1 EtOAc/hexanes) gave the title compound (453.5 mg, 26%) as alight yellow oil: TLC Rf (1: 1 EtOAc/hexanes) 0.44, MS (ES) m/e 364.2 (M + H)+.
g) Ethyl 4-(4-carboxyphenyl)-3-(dimethylaminocarbonyl)b~lt:~noz-tf~
TFA (2.3 mL) was added all at once to a solution ethyl 4-[4-(tert-butoxycarbonyl)phenyl] 3-(dimethylaminocarbonyl)butanoate (168.6 mg, 0.46 mmole) in anhydrous CH2Cl2 (2.3 rnL) at 0C. The solution was stirred at RT for 0.5 hr, then was concentrated to dryness on the rotavap. The residue was reconcentrated from toluene to afford the title colllpoulld as a light yellow oil: MS (ES) mle 308.0 (M + H)1.
h) Ethyl (+)-4-[4-t[[(ben7 i ~ I .ifl~7ol-2-yl)methyl]methylamino~call,onyl]phenyl}-3-(dimethylaminocarbonyl)bl~t~ o~t~
EDC (105.8 mg, 0.55 mmole) was added to a solution of ethyl 4-(4-carboxyphenyl)-3-(dimethylaminocarbonyl)butanoate (0.46 mmole), 2-(methylaminomethyl)ben7imicl~7ole dihydrochloride (129.2 mg, 0.55 mmole), - 35 HOBt ~ H2O (74.6 mg, 0.55 mmole), and diisopropylethylamine (0.32 mL, 1.84 mmole) in anhydrous CH3CN (2.3 mL) at RT. After 22 hr, the reaction was concentrated, and the yellow residue was chromatographed on silica gel (10%
W O 97/24119 PCTrUS96/20748 MeOH in 1:1 EtOAc/CHCl3). The title compound (191.5 mg, 92%) was obtained as a light yellow oil: TLC Rf (10% MeOH in 1:1 EtOAc/CHCl3) 0.44; MS
(ES) m/e 451 (M + H)+.
S i) (V-4-[4-[r[(l~en7.imicl~7.ol-2-yl)methyl]methylamino]carbonyl]phenyl]-3-~dimethylaminocarbonyl)butanoic acid A solution of ethyl (i)-4-[4-[[[(benzirnidazol-2-yl)methyl]methylamino]carbonyl]phenyl]-3-(dimethylaminocarbonyl)butanoate (191.5 mg, 0.43 mmole) and 1.0 N LiOH (0.52 mL, 0.52 mmole) in T~IF (2.2 mL) and H~O (1.6 mL) was stirred at RT for 17 hr, then was acidified with TFA (0.10 rnL, 1.29 mmole). Concentration left an aqueous residue which was purified by ODS chromatography (17~o CH3CN/H2O cont:~inin~ 0.1% TFA; chromatographed againusingl5%CH3CN/H2Ocont~ining0.1%TFA). Concentrationand lyophilization gave the title compound (133.4 mg, 4791'o) as a colorless powder:HPLC (PRP-1(~), 20% CH3CN/H20 cont:lining 0.1% TFA) K' = 1.3; MS (ES) m/e 423.2 (M + H)+. Anal. Calcd for C23H26N,O4 ~ 2 CF3CO2H ~ 0.5 H2O: C, 49.17; H, 4.43; N, 8.49. Found: C, 49.13; H, 4.62; N, 8.52.
T~x~ ?le S
Preparation of (S)-2.3.4.5-tetrahydro-7-rrr(ben7imi~7ol-2-yl)mPthyllmethylaminolcarbonyll-4-mPthyl-3-oxo-lH-1 .~ben70diazepine-2-acetic :~rid. r(2.2--limethyl-2-methoxyacetyl)oxy~methyl ester a) (S)-2,3,4,5-Tetrahydro-7-[r[[1-(tert-butoxycarbonyl)bc~ 7Q1-2-yl]methyl]methylamino]carbonyl]-4-methyl-3 -oxo- 1 H- 1 ,4-benzodiazepine-2-acetic acid To a ~ Lulc of (S)-2,3,4,5-tetrahydro-7-r[[(ben7imi~ 7ol-2-yl)methyl~methylamino]carbonyl3-4-methyl-3-oxo- I H- l ,4-benzodiazepine-2-acetic acid (444 mg, 1.0 mmol), triethylamine (0.1464 mT.,l.OS mmol) in DMF (8 mL), was added dropwise di-tert-butyl dicarbonate (230 mg, 1.05 mmol) in DMF (2 mL).
The reaction ll~i~lule was stirred at RT for 18 h. An aliquot was assayed to inrii~ ~te.
only 50% conversion. Another quantity of triethylamine and di-tert-butyl dicarbonate were added and stirring was continued for another 18 h. An aliquot still in-liC~tefl some unreacted material, so a third quantity of reagents was added and the reaction was stirred for another 18 h. The reaction mixture was concentrated to dryness and the residual oil was triturated with water, filtered and vacuum dried at W O 97/24119 PCTrUS96/20748 40-50~C, to give a white solid of the title compound ( 0.442 g, 85%). MS
fES) m/e 522.4 ~M+H]+.
b) (S)-2,3,4,5-Tetrahydro-7-[[l[l-(tert-butoxycarbonyl)ben7imi~l~7ol-2-yl]methyl]methylamino]carbonyl]-4-methyl-3-oxo- lH- 1,4-benzodiazepine-2-acetic acid, ~(2,2-dimethyl-2-methoxyacetyl)oxy]methyl ester To a solution of the compound of Example Sa, (0.209 g, 0.4 mmol) in dIy acetone (10 mL) was added anhydrous polassiulll carbonate (0.25 g, 1.8 mmol). The reaction mixture was stirred at RT and under argon for 1 h. 2-methoxy-2-methylpropanoic acid chloromethyl ester, US Patent 4,602,012, July 22, 1986, (0.334 g, 2.0 mmol) was then added, followed by tetrabutylammonium iodide (0.03 g, 0.08 mmol). The reaction was stirred at RT under argon for 48 h. It was then filtered and the filtrate was concentrated to a yellow oily residue of the titlecompound (0.67g, q~nti~ative yield). TLC RF 0.48 (silica gel, 6% methanol in methylene chloride). MS ~ES) m~e 652.2 [M+H]+.
c) (S)-2,3,4,5-Tetrahydro-7-[[[(ben7imicl~7ol-2-yl)methyl]methylamino]carbonyl]-4-methyl-3 -oxo- 1 H- 1,4-benzodiazepine-2-acetic acid, [(2,2-dimethyl-2-methoxyacetyl)oxy]methyl ester To a solution of the compound in Example Sb (0.67 g, 1 mmol) in methylene chloride (5 mL) was added TFA (1 mL). The reaction was stirred at RT under argonfor 4 h. It was concentrated to dryness, and the residue was evaporated with methylene chloride three times to remove TFA traces, to give the title compound (0.4 g, 73%). This was purified on a flash silica column (step gradient, 2-3%
methanol in methylene chloride). The fractions cont:linin~ the pure compound were collected, concentrated to yield the title compound (65 mg ) as an off-white solid.
MS (ES) m/e 552.2 [M+H]+. 'H NMR (400 MHz, (CDCl3) ~ 7.6 (br s, lH), 7.22 (m, 6H), 6.5 ( d, lH), 5.85 ( d, lH), 5.8 (d, lH), 5.4 (d, lH), 5.05 (m, lH), 4.79 (q, 2H), 4.3 (d, lH), 3.7 (d, lH), 3.25 ( s, 3H), 3.15 (s, 3H), 3.05 ( s, 3H), 3.02 ( dd, lH), 2.7 (dd, lH), 1.4 (s, 6H). Anal. Calcd for C~8H33N507 1.25 H20: C, 58.58; H, 6.23; N, 12.20. Found: C, 58.60, H, 5.94, N, 12.00.
Fxam~?le 6 P.~paldlion of fi)-2.3.4.5-tetrahydro-7-rrrfben~imi~ 7Ql-2-yl)methyl~ aminolcarbonyll-4-methyl-3-oxo- 1 H- 1.4-benzodiazepine-2-fN-hydroxy~ fzlmi~l~
W O 97124119 PCT~US96/20748 a) (~)-2,3,4,5-Tetrahydl o-7-[[~(benzimidazole-2-yl)methylamino]call,ollyl]-4-methyl-3-oxo- 1 H- 1,4-benzodiazepine-2-(N-hydroxy)~cetz~
NaOMe (Aldrich, 25 wt. % solution in MeOH, 2.2 mL, 9.7 mmole) was added to a solution of hydroxylamine hydrochloride (0.67 g, 9.7 mmole) in MeOH
(40 mT-) at 45~C, and the ~ Lul~e was stirred for 5 min. Methyl (~-2,3,4,5-tekahydro-7-[[[(kpn7imi~ Ql-2-yl)methyl]amino]carbonyl]-4-methyl-3-oxo-lH-1,4-benzodiazepine-2-acetate (0.82 g, 1.9 mmole) was s-lcpçn~lP-l in MeOH (2 mL)and THF (15 mL) and added dropwise to the above solution. The reaction was then stirred at 45~C for 24 h. The ll~ lul~ was concentrated in vacuo, and was then treated with 10% CH3CN/H20 conl~inin~ 0.1% TFA (5 mL). All material dissolved, and then a solid precipitated out. Half of this material was dissolved in the mobile phase by addition of excess TFA and was purified by. prepd.d~ e HPLC (YMC
ODS-AQ, 50 x 250 mm, flow rate = 80 mL/min, 10% CH3CN/H20 con~z~ining 0.1%
T~A; tR = 57 min) to yield the title compound (91 mg, 22%) as a white solid. MS
(ES) m/e 423.1 [M+H] '. 'H NMR (400 MHz, DMSO-d6) ~ 9.06 (bt, J = 4 Hz, lH), 7.77 (m, 2H), 7.58 (m, 2H), 7.50 (m, 2H), 6.60 (d, J = 10 Hz, lH), 6.40 (bs, lH), 5.52(d,J= l9Hz, IH),5.18(bt,J=9Hz, lH),4.85(d,J=6Hz, lH),3.83(d,J=
19 Hz, lH), 2.95 (s, 3H), 2.60 (dd, J = 17, 9 Hz, lH), 2.28 (dd, J = 15, 7 Hz, IH).
Anal. Calcd for C2,H22N6O" ~ 1.5 C2HF3O2 1.0 H2O): C, 47.14; H, 4.20; N, 13.74.
Found: C, 46.95; H, 4.24; N, 13.37.
F.x~nu?le 7 Preparation of (+)-2.3.4~5-tetrahydro-7-r3-(benzimidazol-2-yl~pher~vll-4-methyl-3-oxo- 1 H- 1 ~4-benzodiazepine-2-acetic acid a) 2-(3-Iodophenyl)benzilllidazole To a cold solution of 3-iodobenzoic acid (5~0 g, 20 mmol) and Et3N (3~7 mL, 26 mmol) in THF (50 mL) was added isobutylchloroformate (2~9 mL, 21 mmol3.
The solution was stirred for 1 h at 10~C All of the solution was added slowly to a solution of 1,2-diaminobenzene (2~2 g, 20 mmol) in THF (50 mL)~ After 18 hr, thereaction was concentrated and the residue was partitioned between EtOAc and 5%
Na2CO3~ The layers were separated and the EtOAc layer was washed with water.
Concentration of the organic layer gave a residue which was treated with EtOAc and allowed to stand for 1~ min. Filtration gave a solid which was treated with AcOH(50 rnL) and heated to 110~C After 18 hr, the solution was concentrated. The W O 97/24119 PCTrUS96/20748 residue was treated with EtOAc and the solution was filtered to give the title cc,~ oulld (3.14 g, 50%): MS (ES) m/e 321.2 (M+H)~.
b) 2-[(3-Tributylstannyl)phenyl]l,en,;...icl~7Ole S A solution of 2-(3-iodophenyl)ben7imitl~7Ole (1.0 g, 3.1 mmol), bistributyltin (3.9 mL, 6.2 mmol) and PdCl~(PPh3)2 (100 mg, 0.14 rnmol) in DMF
(10 mL) was heated to 90~C under argon. After 2 hr, the solution was concentrated.
The residue was treated with hexane and filtered. EtOAc was added and the solution was filtered. The filtrate was concentrated to give the title compound (812 mg, 54%): MS (ES) m/e 485.4 (M+H)'.
c) Methyl (i)-2,3,4,5-tetrahydro-1-(tert-butoxycarbonyl)-7-iodo-4-methyl-3-oxo-1 H- 1,4-benzodiazepine-2-acetate To a solution of methyl-7-iodo-4-methyl-3-oxo-lH-1,4-benzodiazepine-2-acetate (1.6 g, 4.3 mmol) and DMAP (10 mg, 0.08 mmol) in CH3CN (10 mL) was added di-tert-butyl dicarbonate (2.0 g, 8.6 mmol), and the solution was stirred at RT.
Additional di-tert-butyl dicarbonate (a total of 8 g, 34.4 mmol) was added periodically until the reaction went to completion. Concentration and silica gelch~ lugraphy gave the title compound (1.8 g, 90%): MS (ES) mle 497.2 (M+Na)' d) Methyl (i)-2,3,4,5-tetrahydro-7-[3-(ben7imi~1~7ol-2-yl)phenyl]- 1 -(tert-butoxycarbonyl)-4-methyl-3-oxo- lH- 1,4-benzodiazepine-2-acetate A mixture of 2-[(3-tributylstannyl)phenyl3ben7imicl~7ole (0.24 g, 0.5 mmol), methyl (+)- 1 -(tert-butoxycarbonyl)-7-iodo-4-methyl-3-oxo- 1 H- 1,4-benzodiazepine-2-acetate (0.223 g, 0.7 mmol), CuI (10 mg, 0.05 mmol), and PdCl2(PPh3)z (40 mg, 0.05 mmol) in DMF (10 mL) was heated to 100~C under argon. After 18 hr, the solution was concentrated. Silica gel chromatography gave the' title compound (0.06 g, 22%): MS (ES) m/e 541.5 (M+H)t.
e) Methyl (_)-2,3,4,5-tetrahydro-7-[3-(be~.~i.. ifl~7ol-2-yl)phenyl]-4-methyl-3-oxo-lH- 1,4-benzodiazepine-2-acetate A solution of methyl (+)-2,3,4,5-tetrahydro-7-[3-(ben7imi~1~7~ 1-2-yl)phenyl]-1 -(tert-butoxycarbonyl)-4-methyl-3-oxo- 1 H- 1,4-benzodiazepine-2-acetate (0.06 g, 0.11 mmol) in 4 M HCl/dioxane (3 mL) was stirred for 1 hr at RT. The solution was concentrated to give the title compound (0.05 g, 100%): MS (ES) m/e 441.4 (M+H)~.
W O 97/24119 PCT~US96/20748 f) (_)-2,3,4,5-Tekahydro-7-[3-(be~ idazol-2-yl)phenyl]-4-methyl-3-oxo-lH-1,4-ben7O~ 7~pine-2-acetic acid 1.0 N NaOH (0.22 mL, 0.22 mmol) was added dropwise to a solution of methyl (+)-2,3,4,5-tetrahydro-7-[3-(b~n7imill~7ol-2-yl)phenyl]-4-methyl-3-oxo-lH-1,4-benzodiazepine-2-acetate (0.05 g, 0.11 mmol) in 1: 1 MeOH/H2O (2 mL) at RT, and the resulting solution was stirred for 18 hr, then was concentrated. The residue was dissolved in H,O and the solution was acidified with AcOH to pH 4 (Litmus paper). Filtration gave the title compound (0.005 g, 10%): 'H NMR (250 MHz, DMSO-d6) o 2.4-2.9 (m, 2H), 3.0-3.1 (s, 3H), 3.8-4.0 (d, lH), 5.0-5.1 (m, lH), 5.5-5.6 (d, lH), 6.7-6.8 (d, lM), 7.5-8.5 (m, 1 lH); MS (ES) m/e 427.5 (M+H)f. Anal.Calcd for C25H"N4O3 1.5 HCl - 1.0 AcOH 0.5 H20: C, 58.94; H, 5.22; N, 10.18.
Found: C, 59.00; H, 5.15; N, 9.92.
Fx~rr~le 8 P1~AI ~lion of (+)-2.3.4.5-tetrahydro-4-methyl-3-oxo-7-r~rtph~ n~ l ., hnidazol-2-yl)m~ thyll"minolcarbonyll- 1 H- 1.4-benzodiazepine-2-acetic acid a) 2-[[(N-Benzyloxycarbonyl)amino]methyl]phen~nthrimi~1~7Ole Following the general procedure of Exarnple 7(a), except substituting N-Cbz-glycine for the 3-iodobenzoic acid, and 9,10-tliz~minophen~nthrene for the 1,2-diaminoben_ene, the title compound (0.41 g, 45%) was prepared: MS (ES) m/e 382.4 (M+H)i.
b) 2-(Aminomethyl)phen"nthrimi(1~7Ole A solution of 2-t[(N-benzyloxycarbonyl)amino~methyl]ph~ ~nthrimidazole (0.2 g, 0.52 mmol) in 30% HBr in acetic acid (0.8 mL) was stirred at RT for 1 hr.
The solution was concentrated and the residue was treated with Et2O. Filtration gave the title compound (0.138 g, 80%) as an oily residue: MS (ES) m/e 248.3 (M+H)~.
c) Methyl (_)-2,3,4,5-tetrahydro-4-methyl-3-oxo-7-[[~(phen~nthrimi~z~7ol-2-yl)methyl]amino]carbonyl]- lH- 1,4-benzodiazepine-2-acetate EDC (0.08 g, 0.42 mmol) was added to a solution of 2-(aminomethyl)ph~n:~nthrimi~ le (0.138 g, 0.42 rnmol), methyl ( )-7-carboxy~
methyl-3-oxo-2,3,4,5-tetrahydro-lH-1,4-benzodiazepine-2-acetate (0.123 g, 0.42 mmol), HOBt H~O (0.063 g, 0.42 mmol) and Et,N (0.14 mL, l mmol) in anhydrous ~20 W O 97/24119 PCTrUS96/20748 DMF (5 rnL~ at RT. After 18 hr, the reaction was concentrated, and the residue was partitioned between EtOAc and 5% NaHCO3. The layers were separated and the organic layer was washed with H2O. Drying (NazSO4) and concentration gave the title compound (0.2 g, 90%): MS (ES) m/e 522.4 (M+H)+.
d) (+)-2~3~4~5-Tetrahydro-4-methyl-3-oxo-7-[[[(phpn~nthrim~ 7ol-2 - yl)methyl]arnino]carbonyl]-lH-1,4-bçn7o~ 7~pine-2-acetic acid Following the procedure of Example 7(f), methyl (+)-2,3,4,5-tetrahydro-4-methyl-3-oxo-7-[[[(phen~nthrimi(lzl71>1-2-yl)methyl]amino}carbonyl]-lH-1,4-benzodiazepine-2-acetate (0.2 g, 0.38 mmol) was saponified and purified to give the title compound (0.014 g, 10%): MS (ES) m/e 508.5 (M+H)+. Anal. Calcd for C29H25N5O4 1.0 TFA 3.0 H2O: C, 55.11; H, 4.77; N, 10.37. Found: C, 55.38; H, 5.13; N, 10.74.
F.x~nl~l?le 9 Ple~)a dlion of methyl (+)-7-carboxy-4-(2.2~2-trifluoroethyl)-3-oxo-2.3.4.5-tetr:~ydro- 1 H- 1.4-benzodiazepine-2-acetate a) tert-Butyl 3-[(2,2,2-trifluoroethyl)arnino]methyl-4-nitroben7O~t~
tert-Butyl 3-bromomethyl-4-nitrobenzoate (2.4 g, 8 mmol)was dissolved in dry THF (50 mL), and 2,2,2-trifluoroethylamine (3 rnL, 38 mmol) was added all atonce. The orangish-yellow solution was stirred at RT for 40 min, then was concentrated to remove the TH~. The residue was diluted with Et20 (100 rnL) and washed twice with 10 % aqueous Na2CO3 (50 mL) and brine (50 rnL). The organic layer was dried (MgSO4). Concentration and silica gel chromatography (2.5% -10% EtOAc/hexanes) gave the title compound (1.6 g, 63%) as a yellow oil: IH
NMR (250 MHz, CDC13) o 8.21 (d, J = 1.3 Hz, lH), 8.03 (dd, J = 8.4, 1.3 Hz, lH),7.96 (d, J = 8.4 Hz, lH), 4.20 (s, 2H), 3.24( q, J = 9.3 E~z, 2H), 1.62 (s, 9H).
b) tert-Butyl 3-[[N-(2,2,2-trifluoroethyl)-N-(tert-buto~yc~l~onyl)3amino]methyl-4-nitro~en7O~te Di-tert-butyl dicarbonate (2.15 g, 10 mmol) was added all at once to a solution of tert-butyl 3-[(2,2,2-trifluoroethyl)amino]methyl-4-nitrobenzoate (1.6 g, 5 mmol) in CH2Cl2 (25 mT.) at RT. The reaction was concentrated and heated to 50~Cunder vacuum for 18 hours. Silica gel chromatography (2% - 5% EtOAc/h~-x~n~s) gave the title compound (2 g, 96%) as a yellow oil: ~H NMR (400 MH_, CDCl3) W O 97/24119 PCTrUS96/20748 7.85-8.15 (m, 3H), 4.75-5.05 (m, 2H), 3.80-4.10 (m, 2H), 1.60 (s, 9H), 1.15-1.80 (m, 9H).
c) tert-Butyl 4-amino-3-[[N-(2,2,2-trifluoroethyl)-N-(tert-butoxycarbonyl)]amino~methylben70~tl ~
10% Pd/C (.4 g, .4 mmol) was added to a solution of tert-butyl 3-[[N-(2,2,2-trifluoroethyl)-N-(tert-butoxycarbonyl)3amino~methyl-4-nitrobenzoate (2.0 g, 5 mmol) in EtOAc (20 mL), and the ~ Lul~ was shaken on a Parr apparatus at RT
under H2 (55 psi). After 4 h, the reaction was filtered through celite(~), and the filtrate was concentrated to afford the title compound (1.9 g, 99%) as a colorless oil:
lH NMR (400 MHz, CDC13) ~ 7.76 (dd, J = 8.5 Hz, 1.8 Hz, lH), 7.68 (d, J = 1.8 Hz, lH), 6.62 (d, J = 8.4 Hz, lH), 4.53 (s, 2H), 3.69 (m, 2H), 1.58 (s, 9H), 1.51(m, 9H).
d) tert-Butyl (+)-4-[2-(1,4-dimethoxy- 1,4-dioxobutyl)anuno]-3-[[N-(2,2,2-trifluoroethyl~-N-(tert-butoxycarbonyl)]amino]methylbenzoate A solution of tert-butyl 4-amino-3-[[N-(2,2,2-trifluoroethyl)-N-(tert-butoxycarbonyl)]amino]methylbenzoate (1.9 g, S mmol) and dimethylacetylene dicarboxylate (0.58 mL, S.S mmol) in MeOH (10 mL) was heated at reflux for 60 rnin, then was cooled to RT. The resulting solution was combined with MeOE~ (20 mL) and 10% Pd/C (0.5 g, .S mrnol), and the mixture was shaken on a Parr apparatus at RT under H2 (50 psi). After 3 h, the reaction was filtered through celite~, and the filtrate was concentrated on the rotavap. The title compound (1.6 g, 62%) was obtained as a faintly yellow oil: IH NMR (40V MHz, CDC13) ~ 7.85 (dd, J = 8.4, 2.0 Hz, lH), 7.68 (d, J = 2.0 Hz, lH), 6.65 (d, J = 8.4 Hz, lH), 6.15 (br s, lH), 4.55~.70 (m, 2H), 4.40 (1/2 AB, J = 15.3 Hz, lH), 3.71 (s, 3H), 3.70 (s, 3H), 3.35-3.50 (m, 2H), 2.95 (dd, J = 16.9, 6.8 Hz, lH), 2.84 (dd, J = 16.9, 6.9 Hz, lH), 1.56 (s, 18H).
e) Methyl (+)-7-carboxy-4-(2,2,2-trifluoroethyl)-3-oxo-2,3,4,5-tetrahydro-lH-1,4-benzodiazepine-2-acetate TFA (7 mL) was added all at once to a solution of tert-butyl (+)-4-[2-(1,4-~lim~thoxy- 1,4-dioxobutyl)amino}-3-[[N-(2,2,2-trifluoroethyl)-N-(tert-butoxycarbonyl)]amino]methylbenzoate (1.6 g, 3 mmol) in anhydrous CH2C12 (20 rnL) at 0~C, and the faintly yellow solution was warmed to RT. After 2 h, the solution was concentrated on the rotavap, and the residue was reconcentrated from toluene (to remove residual TFA). The reSlllting oil was combined with toluene (10 mL) and Et3N (2 mL, lS mmol), and the mixture was heated to reflux under argon.
After 18 hr the solution was allowed to cool and was concenll~ted to dryness under W O 97/24119 PCT~US96120748 vacuum. The residue was dissolved in a miniml~m of MeOH (ca. 15 mL) at reflux, diluted with H2O ~10 ml), and acidified with glacial AcOH (4 drops). The mixture was kept in the refrigerator overnight then was filtered. The solid was dried under high vacuum to afford the title compound (0.80 g, 76%) as a tan powder: lHNMR (400 MHz, DMSO-d6) o 7.61 (2, lH), 7.57 ~dd, J = 8.5, 2 Hz, lH), 6.63 (d, J =
2 Hz, lH), 6.59 (d, J = 8.5 Hz, lH), 5.59 (d, ~ = 16.7 Hz, lH), 5.25 (m, lH), 4.28 (m, 2H), 4.15 (d, J = 16.7 Hz, lH), 3.61 (s, 3H), 2.86 (dd, J = 16.8, 8.7 Hz, lH), 2.74 (dd, J = 16.8, 5.4 Hz, lH).
f) Methyl (+)-2,3,4,5-tetrahydro-7-[[~(benzimidazol-2-yl)methyl]amino]carbonyl]-3-oxo-4-(2,2,2-trifluoroethyl)- 1 H- 1 ,4-bçn70~ 7epine-2-acetate EDC (0.16 g, 0.86 mmol) was added at RT to a solution of methyl (+)-7-carboxy4-(2,2,2-trifluoroethyl)-3-oxo-2,3 ,4,5-tetrahydro- 1 H- 1 ,4-benzodiazepine-2-acetate (0.20 g, 0.71 mmol), HOBt ~ H2O (0.12 g, 0.86 mmol), 2-aminomethylb~n7imill~701e dihydrochloride (0.19 g, 0.86 mmol), DIEA (0.5 mL, 2.8 mmol), and acetonitrile (5 mL) under argon. The rçslllting solution was stirred at RT overnight, then was concentrated. The residue was partitioned between ethyl acetate and water, and the layers were separated. The organic phase was washed with brine, dried (MgSO4), and concentrated. Silica gel chromatography ~1% - 10%CH30H in CH2Cl~) gave the title compound (0.12 g, 44%) as a tan solid: NMR (400 MHz, DMSO-d6) o 8.59 (t, J = 5 Hz, lH), 7.61 (m, 2H), 7.50 (m, 2H), 7.16 (m, 2H), 6.57 (d, J = 11.1 Hz, lH), 6.17 (d, J = 5 Hz, lH), 5.53 (d, J = 16.7 Hz, lH), 5.13 (m, lH), 4.75 (m, 2H), 4.10 (m, 2H), 3.62 (s, 3H), 2.94 (dd, J = 16.8, 8.5 Hz, lH), 2.69 (dd, J = 16.8, 5.4 Hz, lH).
g) (+)-2,3,4,5-Tetrahydro-7-[[[(bçn7imi~1~7ol-2-yl)methyl]amino~carbonyl]-3-oxo-4-(2,2,2-trifluoroethyl)- 1 H- 1 ,4-benzodiazepine-2-acetic acid A solution f methyl(f)-2,3,4,5-tetrahydro-7-[[[(bçn7imi~l~7ol-2-yl)methyl]amino]carbonyl]-3-oxo4-(2,2,2-trifluoroethyl)- lH- 1 ,4-ben7o~ 7~pine-2-acetate (0.12 g, 0.25 mmol) and lithium hydroxide monohydrate (0.017 g, 0.4 mmol) in THF (10 mL), CH30H (2 mL), and H2O (2 mL) was stirred at RT overnight. It was then concentrated and the residue was dissolved in water. The solution was brought to pH 4 with 3 N HCI, then was refrigerated for 1 hour. The resulting solid was collected by filtration and dried to give the title co~ oul-d (0.11 g, 90%) as a ~ 35 white solid: Ms (ES) m/e 476 ~M+H]~. Anal. Calcd for C22H2oN5F304 1.25 H20: C, 53.07; H, 4.55; N, 14.06. Found: C, 52.85; H, 4.36; N, 13.98.
W O 97/24119 PCT~US96/20748 Example 10 Preparation of f+)-2.3.4.5-tetr~ dro-7-rrrbenzimidazol-2-yl)methyllmethylaminolcarbon,yll-3-oxo-4-(2.2.2-trifluoroethyl)- 1 H- 1.4-S berl7~ 7Ppine-2-acetic acid a) Methyl (+)-2,3,4,5-tetrahydro-7-[[[ben7imi(1:-7ol-2-yl)methyl]methylamino]carbonyl~-3-oxo-4-(2,2,2-trifluoroethyl)- I H- 1,4-benzodiazepine-2-acetate Using the procedures of Example 9(f), except substituting 2-(methylaminomethyl)bçn7imic~7ole dihydrochloride for the 2-aminomethylbenzimidazole dihydrochloride, the title compound was prepared: 'H
NMR (400 MHz, CDCl3) ~ 7.67 f~m, 2 H), 7.37 (m, 2 H), 7.25 (m, 2 H), 6.54 (d, J =
8 Hz, 1 H), 5.46 (d, J = 16.7 Hz, 1 H), 5.20 (m, 1 H), 5.04 (s, 2 H), 4.71 (m, 1 H), 4.17(m,1H),3.94(m,1H),3.92(d,J=16.7,1H),3.74(s,3H),3.23(s,3H),2.98 (m, 1 H), 2.74 (m, 1 H).
b) (i)-2,3,4,5-Tetrahydro-7-[trbçn7imiti~7ol-2-yl)methyl]methylamino]carbonyl]- -3-oxo-4-(2,2,2-trifluoroethyl)- 1 H- 1,4-benzodiazepine-2-acetic acid Using the procedure of Example 9(g), methyl (+)-2,3,4,5-tetrahydro-7-[[rbenzimidazol-2-yl)methyl]methylamino]carbonyl] -3-oxo-4-(2,2,2-trifluoroethyl)-lH-1,4-benzodiazepine-2-acetate was saponified to afford the title compound: MS
f~ES) mte 490.2 [M+H~+. Anal. Calcd for C23H22N5F30" 2.25 H20: C, 52.12; H, 5.04; N,13.21. Found: C, 52.00; H, 5.12; N, 13.09.
Fx~n~l?le 11 Prep ~ration of (+)-2.3.4.5-tetrahydro-7-rrr(4-aza-S-methylben7.imicl~7ol-2-yl~rnf~tllyllaminolcarbonyll-3-oxo-4-f2.2.2-trifluoroethyl)-lH-1.4-benzodiazepin~-2-zlf~Ptic acid a) Methyl (+)-2,3,4,5-tetrahydro-7-~[r(4-aza-5-methylben7imirl:~7OI-2-yl)methyl]amino]carbonyl]-3-oxo-4-(2,2,2-trifluoroethyl)- 1 H- 1,4-benzodiazepine-2-acetate Using the procedures of Fx:~rnpl~ 9(f~, except ~ubsLiLuLing 2-(aminomethyl)-4-aza-5-methylben7imi~ 7O1e dihydrochloride for the 2-~minom( thylben7imi-l~7oledihydrochloride, the title colllp-)ul~d was prepared: MS (ES) rnte 505.2 ~M + H)+.
W O97124119 PCTrUS96/20748 b) (~ 2,3,4,5-Tetrahydro-7-[[[(4-aza-5-methylben7imi~1~7ol-2-yl)methyl]amino}carbonyl]-3-oxo-4-(2,2,2-trifluoroethyl)- lH- 1,4-b~4n7.o-1iA7epine-2-acetic acid Using the procedure of Example 9(g), methyl (~)-2,3,4,5-tetrahydro-7-[[[(4-aza-5-methylbenzimidazol-2-yl)methyl]amino]carbonyl]-3-oxo-4-(2,2,2-~ trifluoroethyl)-lH-1,4-benzodia~epine-2-acetate was saponified to afford the title compound: MS (ES) m/e 491.2 (M+H)+. Anal. Calcd for C~H2lN6F3O4 - 2 7/8 H2O:
C, 48.73; H,4.97; N, 15.50. Found: C, 48.50; H, 4.59; N, 15.33.
F.x~n~le 12 al~lion of (+)-2.3.4.5-tetrahydro-7-rrr(ber~7imi(1~7ol-2-yl)methyllmethylamino]carbonyll-4-r2-(3.4-methylenedioxyphenyl)ethyl]-3-oxo-lH-1.4-benzotli~7~-pine-2-acetic acid a) Methyl (+)-2,3,4,5-tetrahydro-7-[[[(ben7imi~ 7~-1-2-yl)methyl3methylamino]carbonyl]-4-[2-(3,4-methylenedioxyphenyl)ethyl]-3-oxo-lH- 1,4-benzodiazepine-2-acetate EDC (0.10 g, 0.55 mmol) was added at RT to a solution of methyl 7-carboxy-4-[2-(3,4-methylenedioxyphenyl)ethyl]-3-oxo- 1 H- 1,4-benzodi~epine-2-acetate (0.14 g, 0.3 mmol), 2-(methylaminomethyl)ben7imicl:~701e dihydrochloride(0.12 g, 0.51 mmole), HOBt ~ H20 (0.072 g, 0.55 mmol), and DIEA ~0.32 mL, 1.84 mmole), in DMF (5 mL) under argon. The resulting solution was stirred at RT
overnight, then was concentrated. The residue was partitioned between ethyl acetate and water, and the layers were separated. The aqueous phase was extracted with ethyl acetate, and the combined organic phases were washed with brine, dried (MgSO4), and concentrated. Silica gel chromatography gave the title compound (0.11 g, 59%) as a colorless foam: 'H NMR (CDCl 3) o 7.62 (m, 2H), 7.31 (m, 2H),7.20 (d, J = 8.1 Hz, lH), 7.07 (s, lH), 6.65 (d, J = 7.9 Hz, lH), 6.60 (s, lH), 6.55 (d, J=7.9Hz, lH),6.46(d,J=8.1 Hz, lH),5.90(d,J=5.4Hz,2H),5.26(d,J= 16.5 Hz, lH), 5.02 (m, lH), 4.93, (d, J = 14.6, lH), 4.83 (d, J = 14.6 Hz, lH), 4.51 (d, J =
5 Hz, lH), 3.74 (s, 3H), 3.71 (m, lH), 3.60 (m, lH), 3.58 (d, J = 16.5 Hz, lH), 3.18 (s, 3H), 2.99 (dd, J = 16, 6.8 Hz, lH), 2.70 (m, lH).
b) (+)-2,3,4,5-Tetrahydro-7-[[[(ben7imi~1~7Ol-2-yl)methyl]methylamino]carbonyl]-4-~2-(3,4-methylenedioxyphenyl)ethyl] -3-oxo- 1 H- 1,4-benzodiazepine-2-acetic acid W O97/24119 PCT~US96/20748 To a solution stirred at RT of methyl (_)-2,3,4,5-tetrahydro-7-[[[(benzirnidazol-2-yl)methyl]methylamino]carbonyl]-4-[2-(3,4-methylenedioxyphenyl)ethyl]-3-oxo-lH-1,4-benzodiazepine-2-acetate (0.11 g, 0.19 rnmol) in THF (1 mL) was added a solution of lithium hydroxide monohydrate (0.01g, 0.23 rnrnol) in H20 (1 mL). The r~s~ ing solution was stirred at RT overnight, then was concentrated to dryness. The residue was dissolved in H2O and the solution was washed with ethyl acetate, then was brought to pH 4 with 3 N HCl.
The res~ ing precipitate was collected by ~lltration and dried to give the titlecompound (0.055 g, 51%) as a white solid. MS (ES) m/e 556.2 [M+H]+. Anal.
Calcd for C3nH29NsO6 H2O: C, 62.82; H, 5.45; N, 12.21. Found: C, 62.69; H, 5.26;N, 12.15.
Fx~ ?le 13 ~lel)a-~lion of (+)-2.3.4.5-tetrahydro-7-rl2-(benzimidazol-2-yl)acetyll~mino]-5-oxo-4-(2-phenylethyl)- 1 H- 1.4-benzodi~epine-2-acetic acid a) Methyl (+)-2,3,4,5-tetrahydro-7-[t2-(be~ idazol-2-yl)acetyl]amino]-5-oxo-4-(2-phenylethyl)- 1 H- 1,4-benzodiazepine-2-acetate EDC (0.27 g, 1.4 rnmole) was added at RT to a solution of (_)-2,3,4,5-tetrahydro-7-amino-5-oxo4-(2-phenylethyl)- 1 H- 1,4-benzodiazepine-2-acetate (0.40 g, 1.1 mmol), ben7imi~1~7Qle-2-acetic acid (Archiv. Der Pharmazie 1960, 293, 758;
0.25 g, 1.4 rnmol), HOBt ~ H[2O (0.20 g, 1.5 mmol), and DIEA (0.35 mL, 2 mmol) in acetonitrile (10 rnL). The res~ ing solution was stirred for 2 days, then was concentrated to dryness. The residue was partitioned between ethyl acetate and water, and the layers were separated. The organic phase was washed with brine, dried (MgSO4), and concentrated. Silica gel chromatography (1% - 10% CH30H in CH2Cll) gave the title compound (0.21 g, 36%) as an amber foam: MS (ES) m/e 512.2 [M+H]+.
b) (+)-2,3,4,5-Tetrahydro-7-t[2-(benzimidazol-2-yl)acetyl]amino]-5-oxo4-(2-phenylethyl)- 1 H- 1,4-benzodiazepine-2-acetic acid A solution of methyl (_)-2,3,4,5-tetrahydro-7-[~2-(ben7.imi~ 1-2-yl)acetyl]amino]-5-oxo4-(2-phenylethyl)- 1 H- 1,4-benzodiazepine-2-acetate (0.21 g, 0.41 mmole) and lithium hydroxide monohydrate (0.022 g, 0.52 mmol) in THF (10 mL) and H20 (2 mL) was stirred at RT overnight, then was then concentrated. The residue was dissolved in water and the solution was washed with ethyl acetate, then W O 97124119 PCT~US96/20748 was brought to pH 4 with 3 N HCl. The resulting precipitate was collected by filtration and dried to give the title compound (0.12 g, 59%) as an off-white solid:
MS (ES) m/e 498.2 [M+H]+. Anal. Calcd for C28H27N5O4 ~ 1.5 H2O: C, 64.11; H, 5.76; N, 13.35. Found: C, 64.36; H, 5.57; N, 13.21.
Exarnple 14 Preparation of (S)-2.3~4.5-tetrahydro-7-rrr(benzimidazol-2-yl)methyllmethylaminolcarbonyl1 -4-methyl-3-oxo- 1 H- 1.4-benzodiazepine-2-acet~mi~l~
a) (S)-2,3,4,5-tetrahydro-7-t[[(bçn7imi~1~7Ol-2-yl)methyl]methylamino]carbonyl]-4-methyl-3-oxo- 1 H- 1,4-benzodiazepine-2-~et~mi~le Methyl (+)-2,3,4,5-tetrahydro-7-[[[(ben ~ 7Ol-2-yl)methyl]amino]callJonyl]-4-methyl-3-oxo-lH-1,4-benzodiazepine-2-acetate (330 mg, 0.76 mmole) in dry MeOH (10 mL) was cooled in an ice bath as ammonia was bubbled into the solution for 0.5 hr. The reaction was then allowed to sit stoppered at RT for 18 hr. After concentration, the residue was purified by silica gel flash chromatography (90: 10 CH2Cl2/MeOH) to give the title compound (52%) as a white solid: MS (ES) m/e 421.2 ~M+H~'. Anal. Calcd for C2~H24N603 1.5 H~O: C, 59.05;
H, 6.08; N, 18.78. Found: C, 58.90; H, 6.04; N, 18.45.
Fx~n~le 15 ~ aldtion of (+)-S-rr2.3.4.5-tetrahydro-7-rrr(benzimidazol-2-yl)methyllamino~carbonyll-3-oxo4-(2-phenylethyl)- 1 H- 1.4-benzodiazepin-2-yllm~thylltetrazole a) Methyl (+)-2,3,4,5-tetrahydro-7-(tert-butoxycarbonyl)-3-oxo-4-(2-phenylethyl)-lH-l,~benzodiazepine-2-acetate Methyl (+)-2,3,4,5-tetrahydro-7-carboxy-3-oxo~-(2-phenylethyl)- 1 H- 1,4-b~n70~ 7~pine-2-acetate (1.0 g, 2.6 mmole) was suspended in toluene (10 mL), andN,N-dimethylformz-micle-di-tert-butyl acetal (5 mL, 20.8 mmole) was added dropwise. The reaction mixture was heated at 80~C for 1.5 hr, then was cooled toRT and poured into 5% Na~CO3 solution. The layers were separated, and the aqueous was extracted with toluene (2 x). The combined organic layers were W O 97/24119 PCT~US96/20748 washed with brine, dried over MgS04, filtered and evaporated to give the title compound (0.9lg, 82%). MS (ES) m/e 439.2 [M+H]+.
b) (i)-2,3,4,5-Tetrahydro-7-(tert-butoxycarbonyl)-3-oxo-4-(2-phenylethyl)- lH- 1,4-benzodiazepine-2-acetic acid A solution of methyl (i)-2,3,4,5-tetrahydro-7-(tert-butoxycarbonyl)-3-oxo-4-(2-phenylethyl)-lH-1,4-benzodiazepine-2-acetate (1.5 g, 3.4 mmole) in ethylene glycol dimethyl ether (160 mL) was treated with H2O (20 mT ) and 0.91 N NaOH (5 mT.). The reaction was stirred under argon at RT for 24 hr, then was acidified to pH
3 with glacial AcOH, concentrated to a small volume (10 mL), and poured into iceH2O. The precipitated solid was collected and dried giving the title compound inqll~ntit~tive yield. MS (ES) m/e 425.2 ~M+H]+.
c) (+)-2,3,4,5-Tetrahydro-7-(tert-butoxycarbonyl)-3-oxo-4-(2-phenylethyl)- lH- 1,4-1 5 benzodiazepine-2-[N-(2-cyanoethyl)~cet~mide]
A solution of (i)-2,3,4,5-tetrahydro-7-(tert-butoxycarbonyl)-3-oxo-4-(2-phenylethyl)-lH-1,4-benzodi~epine-2-acetic acid (1.2 g, 2.6 mmole) in dry DMF
(12 mL) under argon was treated with diisopropylethylamine (0.65 g, 5 mrnole), EDC (0.764 g, 4 rnmole) and HOBt ~ H2O (0.54g, 4 mmole). The resulting solution was stirred for 10 min, then was treated with a solution of 3-aminopropionitrilefumarate in dry DMF (2 mL) cont~inin~ diisopropylethylamine (0.85 g, 6.6 lnrnole).
The reaction was stirred under argon ~or 18 hr, then was concentrated to dryness.
The residue was partitioned between H,O and EtOAc, and the layers were separated.
The organic layer was washed with brine, dried over MgSO4, filtered and concentrated. The oily residue was purified by silica gel flash chromatography (98:2 CH2Cl~/MeOH) to give the title colllpoulld (750 mg, 50%): MS (ES) 477.2 [~+H]+.
d) (i)-1-(2-Cyanoethyl)-5-[[2,3,4,5-tetrahydro-7-(tert-butoxycarbonyl)-3-oxo-4-(2-phenylethyl)- lH- 1 ,4-benzodiazepin-2-yl]methyl]tetrazole A solution of ~+)-2,3,4,5-tetrahydro-7-(tert-butoxycalbollyl)-3-oxo~-(2-phenylethyl)-lH- 1 ,4-benzodi~epine-2-[N-(2-cyanoethyl)acet~mide] (750 mg, 1.3 mmole) in dry THF (15 mL) under argon was treated with triphenylphosphine (1.14 g, 4.6 mrnole), trimethylsilyl~ide (0.52 g, 4.6 mmole), and diethyl~odicarboxylate (0.8 mL, 4.6 mmole) at RT under argon. After 50 hr, the reaction was concentrated to dryness, and the residue was purified by silica gel flash chromatography (98.5:1.5 CH2Cl2/MeOH) to give the tit~e co~ ou,ld (0.56g, 86%): MS (ES) m/e 502.2 [M+H]+.
CA 0224l633 l998-06-26 W O97/24119 PCTrUS96/20748 e) (+)-1-(2-Cyanoethyl)-5-[[2,3,4,5-tetrahydro-7-carboxy-3-oxo-4-(2-phenylethyl)-lH-1,4-benzodiazepin-2-yl]methyl]tetrazole A solution of (+)-1-(2-cyanoethyl)-5-[[2,3,4,5-tetrahydro-7-(tert-butoxycarbonyl)-3-oxo-4-(2-phenylethyl)- 1 H- 1,4-benzodi~epin-2-yl~methyl]tetrazole (0.5 g, 1 mmole) in CH2Cl2 (20 mL) at RT under argon was treated with 4 M HCl in dioxane (10 mL). After 20 hr, the reaction was concentrated to dryness, and the residue was diluted with 5% Na2CO3. The solution was extracted with EtOAc, and the ELOAc layer was discarded. The aqueous layer was acidified with dil HCl and extracted with EtOAc (3x). The combined EtOAc extracts were washed with brine, dried over MgSO4 and evaporated to yield the title compound (0.36g, 81%): MS(ES) m/e 445.4 [M+H]+.
f) (+)-1-(2-Cyanoethyl)-5-[[2,3,4,5-tetrahydro-7-[[[(b~n7imi~1~7ol-2-yl)methyl]amino]carbonyl] -3 -oxo-4-(2-phenylethyl)- 1 H- 1,4-benzodiazepin-2-yl]methyl]tetrazole A solution of (+)-1-(2-cyanoethyl)-5-[[2,3,4,5-tetrahydro-7-carboxy-3-oxo-4-(2-phenylethyl)-lH-l,~ben7ofli~7epin-2-yl]methyl]tetrazole) (360 mg, 0.8 rnmole)dry DMF (5 mL) was treated under argon with DIEA (129 mg, 1 ~nmole), EDC (172 mg, 0.9 mmole) and HOBt ~ H2O (122 mg, 0.9 mmole). The reaction was stirred at RT for 10 min, then a solution of 2-aminomethylben7imit~701e dihydrochloride hydrate (352 mg, 1.6 mmole) in DMF (2 mL) cont:lining DIEA (413 mg, 3.2 mmole) was added. After 20 hr, the reaction was concentrated to dryness and the residue was partitioned between }~2O and EtOAc. The layers were separated, and the aqueous layer was extracted with EtOAc. The organic layers were combined, dried over MgSOt, filtered and concentrated. The residue was purified by silica gel flash chromatography (95:5 CH2Cl2/MeOH) to give the title compound (120 mg, 21%):
MS (ES) m/e 575.2 [M+H]+.
g) (+)-S-[r2,3,4,5-Tetrahydro-7-[[[(b~ 7-)1-2-yl)methyl]amino]carbonyl]-3-oxo~-(2-phenylethyl)- l H- 1,4-benzodiazepin-2-yl]methyl]tetrazole A solution of (+)-1-(2-cyanoethyl)-5-[[2,3,4,5-tetrahydro-7-[[[(ben7imi~ 7Ol-2-yl)methyl]amino]carbonyl]-3-oxo-4-(2-phenylethyl)- I H- 1,4-benzodiazepin-2-yl]methyl]tetrazole (100 mg, 0.2 mmole) in MeOH (1 mL) was treated with ~ 35 thiophenol (0.02 mL) followed by 1 N NaOH solution (2.2 mL). After 3 hr, the reaction was concentrated to dryness, and the residue was purified by silica gel~pa,a~ive TLC (85: 15 CH2Cl2/MeOH). The isolated product was dissolved in HzO, W O 97/24119 PCT~US96/20748 and the solution was filtered to remove insoluble m~t~ri~l~ The filtrate was treated with 2 drops of glacial AcOH. The precipitated solid was collected and dried to give the title compound (45 mg, 41%): MS (ES) m/e 522.2 ~M+H]'. Anal.
Calcd for C3~ Ng04 ~ 2.25 H2O: C, 56.15; H, 5.52; N,19.65. Found: C, 56.51; H, 5.05; N, 19.72.
~x~rn,ple 16 Preparation of (S)-2.3.4.5-tetrahydro-7-rrrfbe~ .idazol-2-10 yl~methyllaminolcarbonyll-3-oxo4-r4-r(2-carboxybenzoyl)aminolbut-1-yll-lH-1.4-ben70~ pine-2-acetic acid a) N-[[2-(N-4-hydroxybut- 1-yl)aminomethyl-4-tert-butoxycarbonylJphenyl~-L-aspartic acid ,l3-methyl ester A mixture of N-[[2-formyl-4-tert-butoxycarbonyl]phenyl]-L-aspartic acid ,13-methyl ester (WO 95/18619; 2.55g, 7.26 mmol), 4A molecular sieves, and 4-hydroxybutylamine (0.64 g, 7.26 mmol) in MeOH (35 mL) was stirred under argon at RT for 30 min, then sodium cyanoborohydride (0.49 g, 0.79 mmol) and acetic acid (0.3 mL) were added. The reaction mixture was kept at RT overnight and then20 the solvent was elimin~te~l in vacuo. The residue was dissolved in H20 and the solution was acidified to pH 4 with dil HCl. EtOAc extraction, drying (MgSO4), filtration, and conce~ dLion gave the title compound ( 1.75 g, 57%) as a pale yellow solid: TLC Rf (4:20:20:56 MeOH/EtOAc/hexane/Cl2CH~) 0.22; 1~ NMR (CDCl3) o 1.55 (s, 9H), 1.56 (m, 2H), 1.80 (m, 2H), 3.01 (m, 4H), 3.55 (m, 2H), 3.70 (s, 3H), 25 4.05 (m, lH), 4.40 (m, lH), 4.55 (m, lH), 6.81 (d, J = 8.4 Hz, lH), 7.70 (s, lH), 7.89 (d, J = 8.4 Hz, lH).
b) Methyl (S)-7-(tert-butoxycarbonyl)-2,3,4,5-tetrahydro-3-oxo-4-(4-hydroxybut- 1 -yl)- 1 H- 1,4-benzodi~epine-2-acetate To a solution of N-[[2-(N-4-hydroxybut-1-yl)aminomethyl-4-tert-butoxycarbonyl]phenyl]-L-aspartic acid ,B-methyl ester (1.75 g, 4.1 mmol) and triethylamine (1.15 mL, 8.2 mmol) in dichloromethane (150 mL) under argon at RT
was added benzotriazol- 1 -yloxy-tris(dimethylamino)phosphonium hexafluorophosphz-~ (2.08 g, 14.7 mmol). The reaction mixture was stirred 35 overnight at RT, then was washed sequentially with ice-cold dil HCl, water, 5%
sodium bicarbonate, saturated brine, and then dried (MgS04). Filtration and concentration left a residue which was purified by silica gel flash column W O 97/24119 PCT~US96/20748 chromatography (5% mPth~nol:ethyl acetate) to give the title compound (0.631 g, 38%): TLC Rf (4% MeOH/EtOAc) 0.26; IH NMR (CDC13) o 1.46-1.61 (m, 4H), 1.57 (s, 9H), 2.64 (d, J = 6.9 Hz, lH), 2.66 (dd, J = 15.9, 6.3 Hz, lH), 2.99 (dd, J = 15.6, 6.9 Hz, lH), 3.56-3.54 (m, 4H), 3.74 (s, 3H), 3.84 (d, J = 16.2 Hz, lH), 4.54 (m, lH), 5.10 (m, lH), 5.41 (d, J = 16.2 Hz, lH), 6.49 (d, J = 8.3 Hz, lH), 7.59 (d, J = 1.8 Hz, lH), 7.67 (dd, J = 8.3, 1.8 Hz, lH); MS (ES) m/e 407.2 [M+H]+; ~CC~D
= - 185.4~ (c = 1, CH30H).
c) Methyl (S)-7-(tert-butoxycarbonyl)-2,3,4,5-tetrahydro-3-oxo-4-(4-phth~limic~obut-1-yl)-lH-1,4-benzodiazepine-2-acetate To a solution of methyl (S)-7-(tert-butoxycarbonyl)-2,3,4,5-tetrahydro-3-oxo-4-(4-hydroxybut-1-yl)-lH-1,4-benzodiazepine-2-acetate (437 mg, 1.07 mmol) and triphenylphosphine (308 mg, 1.17 mmol) in THF (20 mL) at RT under argon was sequentially added phth~limi~ (173 mg, 1.17 mmol) and diethyl azodicarboxylate (205 mg, 1.17 mmol). The reaction ll~ib~luie was stirred overnight at RT, the solvent was removed, and the residue was purified by silica gel flashcolumn chromatography (4:20:20:56 m~ths~n~ll/ethyl acetatethexane/methylene chloride) to give the tit}e compound (0.430 g, 75%): TLC Rf (4:20:20:56 MeOH/EtOAc/hexane/Cl2CH2) 0.32; IH NMR (CDCl3) o 1.55 (s, 9H), 1.55-1.61 (m, 4H), 2.68 (dd, J = 14.0, 5.7 Hz, 2H), 2.98 (dd, J = 14.0, 6.6 Hz, lH), 3.46-3.64 (m, 4H), 3.71 (s, 3H), 3.85 (d, J = 16.5 Hz, lH), 4.63 (d, J = 4.4 Hz, lH), 5.08 (dd, J =
5.7, 6.6 Hz, lH), 5.37 (d, J = 16.5 Hz, lH), 6.48 (d, J = 8.3 Hz, lH), 7.67 (s, lH), 7.69 (d, J = 8.3, 1.8 Hz, lH), 7.72-7.76 (m, 2H), 7.81-7.86 (m, 2H).
d) Methyl (S)-2,3,4,5-tetrahydro-7-carboxy-3-oxo-4-(4-phth:~limidobutyl)-lH-1,4-benzodiazepine-2-acetate To a solution of methyl (S)-7-(tert-butoxycarbonyl)-2,3,4,5-tetrahydro-3-oxo-4-(4-phth~limidobutyl)-lH-1,4-benzodiazepine-2-acetate (660 mg, 0.89 m~nol) in dichloromethane (20 mL) was added 4 N HCI/dioxane (5 mT, 20 mmol) at RT
under argon. The reaction mixture was stirred for 18 hr. The s~lspen~ion was concentrated to give the title compound as an off-white solid (425 mg, 98%): IH
NMR (CDCl3) ~ 1.55-1.61 (m, 4H), 2.71 (dd, J = 14.1, 6.0 Hz, 2H), 3.01 (dd, J =
14.1, 6.3 Hz, lH), 3.50-3.65 ~m, 4H), 3.75 (s, 3H), 3.89 (d, J = 16.5 Hz, lH), 4.68 - (d, J = 4.5 Hz, lH), 5.12 (dd,3 = 6.0, 6.3 Hz, lH), 5.40 (d, J = 16.6 Hz, lH), 6.41 - 35 (bs, lH), 6.53 (d, J = 8.4 Hz, lH), 7.69-7.75 (m, 4H), 7.82-7.85 (m, 2H); MS (l~S) m~e 480.2 [M+H]+.
W O 97/24119 PCT~US96/20748 e~ Methyl (S)-2,3,4,5-tetrahydro-7-[[[(b~n7imi-1~7Ol-2-yl)methyl3amino]carbonyl]-3-oxo~-(4-phth:~limiclobut-1-yl)-lH-1,4-benzodiazepine-2-acetate EDC (240 mg, 1.25 mmol) was added to a stirred solution of methyl (S)-2,3,4,5-tetrahydro-7-carboxy-3-oxo-4-(4-phth~limi~lQbut-l-yl)-lH-1,4-benzodiazepine-2-acetate (0.85 g, 0.88 mmol), 2-(aminomethyl)'oe~.7i.,~ 7Ole dihydrochloride (230 mg, 1.04 mmol), HOBt -H2O (169 mg, 1.25 mmol), and diisopropylethylamine (0.78 mL, 4.5 mmol) in anhydrous acetonitrile (10 rnL) at RT. After 19 h, the reaction was concentrated on the rotavap (high vacuum), and the residue was partitioned between H20 (5 mL) and EtOAc (20 mL). The layers were separated and the organic layer was washed with H2O (5 rnL). Drying (MgSO4), concentration, and silica gel chromatography (5% MeOH/CH2Cl2), gave the title compound (230 mg, 43%) as an off-white solid: TLC Rf (5% MeOH/Cl2CH2) 0.30;
lH NMR (CD30D) o 1.42-1.56 (m, SH), 2.63 (dd, J = 6.4, 16.2 Hz, lH), 2.95 (dd, J= 6.7, 16.2 Hz, lH), 3.33-3.40 (m, 2H), 3.48-3.55 (m, 2H), 3.57 (d, J = 16.5 Hz, lH), 3.67 (s, 3H), 4.72~.80 (m, 3H), 5.03 (dd, J = 6.4, 6.7 Hz, lH), 5.20 (d, J = 16.5 Hz, lH), 6.44 (d, J = 8.4 Hz, lH), 7.18-7.21 (m, 2H), 7.52-7.63 (m, 6H); 7.74-7.76 (m, 2H), 9.08 (br s, lH).
f) (S)-2,3,4,5-Tetranydro-7-[t[(b~n7imicl~7ol-2-yl)methyl]amino]carbonyl]-3-oxo-4-[4-[(2-carboxybenzoyl)amino]but-1-yl]- lH-1,4-bt-.n7oflizl7~pine-2-acetic acidLiOH (30 mg, 0.71 mmole) was added at RT to a solution of methyl (S)-2,3,4,5-tetrahydro-7-[[[(benzimidazol-2-yl)methyl]amino]carbonyl]-3-oxo-4-(4-phth:~limi~lobut-l-yl)-lH-1,4-benzodiazepine-2-acetate (223 mg, 0.33 mmol) in MeOH (2 mL) and H20(3 mL). The reaction mixture was sti;red at RT for 19 hr.
Acidification with dil HCl to pH 4 and concentration produced a solid. Filtration gave the title compound (145 mg, 66%) as a white solid: ~a]D - -100.4~ (c = 1, CH30H); IH NMR (CD30D) ~ 1.32-1.65 (m, 5EI), 2.58 (dd, J = 16.4, 6.7 Hz, lH), 2.90 (dd, J = 16.4, 7.9 Hz, lH), 3.06 (m, lH), 3.69 (m, lH), 4.00 (d, J = 16.7 Hz, lH),4.69(brs,2H),5.11 (dd,J=7.9,6.7Hz, lH~, 5.36(d,J= 16.7Hz, lH),6.50 (d, J = 8.4 Hz, lH), 7.29 (m, 2H), 7.37 (m, 4H); 7.51 (m, 2H), 7.664 (s, lH), 7.74 (d, J = 6.8 Hz, lH); MS (ES) m/e 613.2 [M+H]+. Anal. Calcd for C32H32N607 -1.5 H2O: C, 60.0g; H, 5.51; N, 113.14. Found: C, 59.77; H, 5.46; N, 12.98.
E~ ?le 17 W O 97/24119 PCTrUS96/207~8 F'lc~aldlion of (+)-7-r3-(benzill,idazol-2-yl)propyll-4-methyl-3-oxo-2.3.4.5-tetrahydro- 1 H- 1.4-benzodiazepine-2-acetic acid a) Methyl (_)- 1 -(tert-butoxycarbonyl)-7-(4-hydroxy- 1 -butyn- 1 -yl)-4-methyl-3-oxo-2,3,4,5-tetrahydro- 1,4-benzodiazepine-2-acetate 3-Butyn-l-ol (65 mg, 0.93 mmol), bis(triphenylphosphine)pS~ m (II~
chloride (5 mg, 0.007 mmol), triphenylphosphine (10 mg, 0.038 mmol), and copper(I) iodide (10 mg, 0.052 mmol) were added under an Ar atmosphere to a solution of methyl (+)-2,3,4,5 -tetrahydro- 1 -(tert-butoxycarbonyl)-7-iodo-4-methyl-3-oxo-lH-1,4-benzodiazepine-2-acetate (obtained as in example 7c; 440 mg, 0.94 mmol) in triethylamine (34 m~ ). The reaction mixture was heated to reflux for 4 hr, then was filtered through celite(~), and the filtrate was concentrated. The residue was purified by silica gel flash column chromatography (4:20:20:56 MeOH/EtOAc/hexane/CI2CH2) to give the title compound (390 mg, g4%) as a pale yellow liquid: TLC Rf (5% MeOH: Cl2CH2) 0.37; IH NMR (400 MHz, CDCl3) o 7.34 (dd, J = 1.8, 8.1 Hz, lH), 7.31 (d, J = 1.8 Hz, lH), 7.13 (d, J = 8.1 Hz, lH), 5.15-5.23 (m, lH), 4.75 (d, J = 14.4 Hz, lH), 3.68 (d, J = 14.4 Hz, lH), 3.63-3.67 (m, 2H), 3.59 (s, 3H), 3.04, (s, 3H), 2.88 (dd, J = 5.5, 15.2 Hz, lH), 2.45 (t, J = 6.4 Hz, 2H), 2.26 (dd, J = 9.5, 15.2 Hz, lH), 2.04 (br s, lH), 1.34 (br s, 9H); MS (ES) m/e 417 [M+H]+.
b) Methyl ( )- 1 -(tert-butoxycarbonyl)-7-(4-hydroxybut- 1 -yl)4-methyl-3-oxo-2,3,4,5-tetrahydro- 1,4-benzodiazepine-2-acetate 10% Pd/C (40 mg) was added to a solution of methyl (+)-l-(tert-butoxycarbonyl)-7-(4-(hydroxy- 1 -butyn- 1 -yl)-4-methyl-3-oxo-2,3,4,5-tetrahydro-1,4-benzodiazepine-2-acetate (370 mg, 0.89 mmol) in EtOH (20 rnL), and the mixture was shaken on a Parr apparatus at RT under H2 (50 psi). After 12 h, the reaction was filtered through celite~, and the filtrate was concentrated to afford the title compound (350 mg, 94%) as a pale yellow liquid: TLC Rf (4:20:20:56 MeOH/EtOAc/hexane/CI2CH2) 0.55; IH NMR (400 MHz, CDCl3) o 7.10-7.19 (m, 3H), 5.59-5.77 (m, lH), 4.85 (d, J = 15.0 Hz, lH), 3.68-3.60 (m, SH), 3.12 (s, 3H), 2.85 (dd, J = 5.4, 15.3 Hz, lH), 2.65 (t, J = 6.4 Hz, 2H), 2.34 (dd, J = 10.0, 15.3 Hz, lH), 1.30-1.78 (m, 13H); MS (ES) rn/e 421 LM+H]+.
-35 c) Methyl (_)-l-(tert-butoxycarbonyl)-7-(4-carboxybut-1-yl)-4-methyl-3-oxo-2,3,4,5-tetrahydro- 1,4-benzodiazepine-2-acetate ~33 W O 97/24119 PCTrUS96/20748 To a solution of methyl (+)-l-(tert-butoxycarbonyl)-7-(4-hydroxybut- I -yl)4-methyl-3-oxo-2,3,4,5-tetrahydro- l ,4-ben7orli~7epine-2-acetate (350 mg, 0.82 mmol) in CH2Cl2 at 0~C was added 2,2,6,6-tetrarnethyl-oxopiperi~linillm chloride (J. Org. C*em. 1985, 50, 3930-3931; 220 mg, 1.1 rnmol).
S The mixture was stirred for 2 hr at 0~C under Ar atmosphere. 2-Methyl-2-butene (1 mL) was added, followed by the addition of a freshly prepared solution of NaClO2~0.76 g, G.7 mmol), NaH2PO, ~ H20 (0.78 g, 5.68 mrnol) and H20 (25 mL). The cooling bath was removed, and the rnixture was taken up in EtOAc and washed successively with 0.05 M HCl and brine. Drying (MgSO4), concentration, and silica gel chromatography (5% AcOH in 4:20:20:56 MeOH/EtOAc/hexane/Cl2CH2), gave the title compound (350 mg, 98%): TLC Rf (5% AcOH in 4:20:20:56 MeOH/EtOAc/hexane/Cl~CH2) 0.32; IH NMR (400 MHz, CDC13) o 7.13-7.18 (m, 3H), 5.60-5.69 (m, lH), 4.83 (d, J = 14.2 Hz, lH), 3.76 (d, J = 14.2 Hz, lH), 3.66 (s, 3H), 3.12 (s, 3H), 2.93 (dd, J = 4.5, 15.3 Hz, lH), 2.67 (t, J = 7.3 Hz, 2H), 2.36 (t, J
= 7.3 Hz, 2H), 2.30-2.34 (m, lH), 1.95 (q, J = 7.3 Hz, 2H), 1.34 (s, 9H); MS (ES) m/e 435 [M+H]+.
d) Methyl (+)-l-(tert-butoxycarbonyl)-7-t3-(~ 7.ol-2-yl)propyl]-4-methyl-3-oxo-2,3,4,5-tetrahydro- 1,4-benzodiazepine-2-acetate To a stirred and cooled (-10~C) rnixture of methyl (+)-l-(tert-butoxycarbonyl)-7-(4-carboxybut- 1 -yl)-4-methyl-3-oxo-2,3,4,5-tetrahydro- 1,4-benzodiazepine-2-acetate (350 mg, 0.8 mmol) and Et3N (81 mg, 0.8 rnrnol) in anhydrous THF (8 mL) was added isobutylchloroformate (97 mg, 0.8 mrnol). After 10 min, a solution of 1,2-phenylenefli:~min~ (1.43 g, 0.9 mmol) in THF (2 rnL) was added. Stirring was continued at RT overnight, then the solvents were evaporated.
The residue was dissolved in EtOAc, and the solution was washed seqllentiz3lly with aqueous NaHCO3 and brine. Drying (MgSO4) and concentration produced a pale yellow solid. This was dissolved in glacial AcOH (5 rnL), and the reaction was heated to 60~C. After 3 hr, the mixture was cooled, concentrated, neutralized with 2.5 N NaOH, and extracted with CH2C12. Drying (MgSO4), concentration, and silica gel chromatography (gradient 1-5% MeOH/CH2C12) gave the title compound (200 mg, 50%): TLC Rf (4:20:20:56 MeOH/EtOAc/hexane/Cl2CH2) 0.18; lH NMR
(400 MHz, CDC13) o 7.57-7.61 (m, 2H), 7.15-7.28 (m, 4H), 7.08 (s, lH), 5.60-5.55(m, lH), 4.75-4.88 (m, lH), 3.71 (d, J = 14.2 Hz, lH), 3.70 (s, 3H), 3.10 (s, 3H), 2.94 (dd, J = 4.2, 15.3 Hz, lH), 2.85-2.90 (m, 2H), 2.73-2.79 (m, 2H), 2.32-2.36 (m, lH), 2.15-2.23 (m, lH), 1.34 and 1.55 (br s, rotamers, 9H); MS (ES) m/e 507 [M+H]+.
W O 97/24119 PCTrUS96/20748 e) (+)-7-[3-(Ben7imi(i~7ol-2-yl)propyl~-4-methyl-3-oxo-2~3~4~s-tetrahydro- lH- 1,4-benzodiazepine-2-acetic acid LiOH (17 mg, 0.71 mmole) was added at RT to a solution of methyl (+)- 1-(tert-butoxycarbonyl)-7-[3-(ben7imi~1~7.ol-2-yl)propyl]-4-methyl-3-oxo-2,3,4,5-tetrahydro-1,4-benzodiazepine-2-acetate (200 mg, 0.395 mmol) in MeOH (2 mL) and H20 (3 mL). The reaction mixture was stirred at RT for 4 hr. Aci~lifi~tion with dil HCl to pH 4 and concentration produced a white solid. This was dissolved in a mixture of methylene chloride (10 mL) and trifluoroacetic acid (5 mL) at 0~C, and the reaction was kept at 0~C for 30 min. The solvents were ~v~olat~d and theresidue was triturated with ether and purified by ODS flash chromatography (gradient 10 to 18% CH3CN/H2O cont~ining 0.1% TFA). Concentration and lyophilization gave the title compound (75 mg, 48%) as a colorless powder: IH
NMR ((400 MHz, CDC13) â 7.58-7.61 (m, 2H), 7.37-7.39 (m, 2H), 6.79 (d, J = 8.2 Hz, lH), 6.68 (s, lH), 6.39 (d, J = 8.2 Hz, lH), 5.18(d, J = 16.8 Hz, lH~, 4.77 (dd, J
= 7.0, 7.5 Hz, lH), 3.63 (d, J = 16.8 Hz, lH), 3.03-3.19 (m, 3H), 2.97 (s, 3H), 2.83 (dd, J = 7.0, 16.4 Hz, lH), 2.52-2.56 (m, 2H), 2.08-2.12 (m, 2H); MS (ES) m/e 393.0 tM+H]~- Anal. Calcd for C22H24N4O3 ~-C2HF3O2 ~ 0.5 H2O: C, 55.92; H, 5.08; N, 10.89. Found: C, 56.16; H, 4.92; N, 10.88.
Example 18 Plc~paldtion of (S)-2.3.4.5-tetrahydro-7-rrN-r(ben7imi~1~7-)1-2-yl)mPtllyl~-N-(4-aminobutyl)aminolcarbonyll-4-methyl-3-oxo- 1 H- 1.4-benzodiazepine-2-acetic acid a) 4-[(B~n7imid~7OI-2-yl)methylJaminobutyronitrile To a stirred mixture of 2-aminomethylben7imi~i~701e dihydrochloride (0.5 g, 2.2717 mmol) and NaHCO3 (0.67 g, 7.951 mmol) in dry DMF (10 mL) was added 4-bromobutyronitrile (0.37 g, 2.4989 mmol). After stirring at RT for 24 hr, the mixture was concentrated. The residue was taken up in H2O and extracted with CH2Cl2. The organic extracts were dried over MgS04, concentrated, and purified by silica gel flash column chromatography (5% MeOH/CH2Cl2)to give the title compound (0.15 g, 35%) as a brown oil: 'H NMR (250 MHz, DMSO-d6) ~ 1.82 (m, - 2H), 2.45 (t, J = 4 Hz, 2H~, 2.85 (t, J = 4 Hz, 2H), 4.11 (s, 2H), 7.14 (m, 2H), 7.50 35 (m, 2H).
-W O 97/24119 PCTrUS96/20748 b) Methyl (S)-2,3,4,5-tetrahydro-7-[[N-[(benzimidazol-2-yl)methyl]-N-(4-cyanopropyl)amino]carbonyl]-4-methyl-3-oxo- lH- 1,4-benzodiazepine-2-acetate To a stirred mixture of 4-~(b~n7im;~ 7Ol-2-yl)methyl]aminobutyronitrile (0.159 g, 0.7422 mmol), methyl 2,3,4,5-tetrahydro-7-carboxy-4-methyl-3-oxo-lH-1,4-benzodiazepine-2-acetate ( 0.217 g, 0.7422 mmol), HOBt H,O (0.120 g, 0.8906 mmol), and i-Pr2NEt (0.192 g, 1.4844 mmol) in dry CH~CN (7 mL) was added EDC
(0.265 g, 0.8906 mmol). After stirring at RT for 48 hr, the ~ lule was concentrated. The residue was taken up in H2O and extracted with CH2Cl2. The organic layer was washed se~uentially with saturated NaHCO3 and brine, dried over MgSO4, and concentrated to give a brown oil. Silica gel flash column chromatography (3% MeOH/CH2CI2) gave the title compound (0.261 g, 74%) as an off white foam: 'H NMR (250 MHz, DMSO-d6): o 1.95 (m, 2H), 2.66 (dd, J = 16.4, 3.5 Hz, lH), 2.78 (dd, J = 16.4, 3.5 Hz, lH), 2.85 (t, J = 8.7 Hz, 2H), 3.45 (t, J = 8.7 Hz, 2H), 3.60 (s, 3H), 3.80 (d, J = 16 Hz, lH), 4.52 (s, 2H), 4.84 (d, J = 2.9 Hz, 2H), 5.15 (m, lH), 5.48 ~d, J = 16 Hz, lH), 6.40 (d, J = 3.5 Hz, IH), 6.54 (d, J = 8.3 Hz, lH), 7.25 (m, 4H);,7.50 (m, lH), 7.62 (m, lH).
c) (S)-2,3,4,5-Tetrahydro-7-[[N-[(ben7imi~1~7.ol-2-yl)methyl~-N-(4-cyanopropyl)amino]carbonyl~-4-methyl-3-oxo- lH- 1,4-benzodiazepine-2-acetic acidTo a stirred solution of methyl (S)- 2t3,4,5-tetrahydro-7-[~N-[(benzimidazol-2-yl)methyl]-N-(4-cyanopropyl)amino]carbonyl]-4-methyl-3-oxo- lH- 1,4-benzodiazepine-2-acetate (0.261 g, 0.5478 mmol) in MeOH (5 mL) was added 2.5 N
NaOH (0.7 mL, 1.6433 mmol). After stirring at RT overnight, the mixture was concentrated. The residue was taken up in H2O, and the solution was acidified with 6 N HC1 to pH = 4. The white solid was filtered and dried to afford the title compound ~0.21 g, 81%): 'H NMR (250 MHz, DMSO-d6): ;~ 1.95 (m, 2H), 2.66 (dd, J = 16.4, 3.5 Hz, lH), 2.78 (dd, ~ = 16.4, 3.5 Hz, lH), 2.85 (t, J = 8.7 Hz, 2H), 3.45(t,J=8.7Hz,2H),3.80(d,J= 16Hz, lH),4.52(s,2H),4.84(d,J=2.9Hz, 2H), 5.15 (m, lH), 5.48 (d, J = 16 Hz, lH), 6.40 (d, J = 3.5 Hz, lH), 6.54 (d, J = 8.3 Hz, lH), 7.25 (m, 4H), 7.50 (m, lH), 7.62 (m, lH~.
d) (S)-2,3,4,5-Tetrahydro-7-[~N-L(benzimidazol-2-yl)methyl]-N-(4-aminobutyl)amino]carbonyl]-4-methyl-3-oxo- lH- 1,4-benzodiazepine-2-acetic acid A mixture of (S)-2,3,4,5-tetrahydro-7-[[N-[(ben~i...i-1~7- 1-2-yl)methyl]-N-(4-cyanopropyl)amino]carbonyll-4-methyl-3-oxo- lH- 1,4-benzodiazepine-2-acetic acid(0.200 g, 0.4325 mmol) and NH40H (1 mL, 30% solution) in MeOH (5 mL) was hydrogenated over Ra/Ni at RT for 24 hr. The catalyst was ~lltered off, and the .
CA 0224l633 1998-06-26 W O97/24119 PCT~US96/20748 filtrate was concentrated and purified by reverse phase chromatography (10% CH3CN/H10 contzlining 0.1% TFA) to give the title compound (0.100 g, 33%) as an off white solid: 'H NMR ( 400 MHz, DMSO-d6) ~ 1.45 (m, 2H), 1.72 (m, 2H), 2.54 (dd, J = 16.4, 3.5 Hz, lH), 2.70 (m, 2H), 2.75 (dd, J = 16.4, 3.5 Hz, lH), 2.95 (s, 3H), 3.65 (t, J = 8.7 Hz, 2H), 3.85 (d, J = 16 Hz, lH), 5.05 (s, 2H), 5.15 (m, lH), 5.48 (d, J = 16 Hz, lH), 6.65 (d, J = 8.3 Hz, lH), 7.20 (m, 2H), 7.61 (m, 2H), 7.75 (s, 2H), 7.85 (m, 2H); IR (KBr) 3425, 3000, 3100, 1728, 1675, 1630, 1625, 1613 cm~';MS (ES) m/e 479 (M+H). Anal. Calcd for C25H3"N604 ~ 2 CF3CO2H: C, 49.30; H, 4.56; N,11.89. Found: C, 49.22; H, 4;89; N, 11.84.
Fx:-ml?le 19 Preparation of ~S)-2.3.4.5-tetrahydro-7-rrN-r(benzimidazol-2-yl)methyl-N-(2-cyanomethyl)amino~carbonyll-4-methyl-3-oxo- lH- 1.4-benzodiazepine-2-acetic acid a) t(Ben7imi(1~7Ol-2-yl)methyl]~minn~retonitrile Following the procedure of Example in 18(a), except substituting bromoacetonitrile for the 4-bromobutyronitrile, the title compound was prepared as an off white solid (0.15 g, 35%): lH NMR (250 MHz, DMSO-d6): o 3.71 (s, 2H), 3.98 (s, 2H), 7.14 (m, 2H), 7.50 (m, 2H).
b) Methyl (S)-2,3,4,5-tetrahydro-7-[[N-[(ben7imicl~7(-1-2-yl)methyl-N-(2-cyanomethyl)amino]carbonyl]-4-methyl-3-oxo- 1 H- 1,4-benzodiazepine-2-acetate Following the procedure of Example 18(b), except substituting [(ben7imi~701-2-yl)methyl]~mino~retQnitrile for 4-l (bçn7imi~1~7ol-2-yl)methyl]aminobutyronitrile, the title compound was prepared as an of ~ white foam (0.487 g, 66%): 'H NMR (250 MHz, DMSO-d6) ~ 2.66 (dd, J--16.4, 3.5 Hz, lH), 2.78 (dd, J = 16.4, 3.5 Hz, lH), 2.92 (s, 2H), 3.60 (s, 3H), 3.80 (d, J = 16 Hz, lH), 4.52 (s, 2H), 4.84 (d, J = 2.9 Hz, 2H), 5.15 (m, lH), 5.48 (d, J = 16 Hz, lH), 6.40 (d, J = 3.5 Hz, lH), 6.54 (d, J = 8.3 Hz, lH), 7.25 (m, 4H), 7.50 (m, 2H), 7.62 (m, 2H).
c) (S)-2,3,4,5-Tetrahydro-7-[[N-[(b~n,; . . ,i~1~7ol-2-yl)methyl-N-(2-cyanomethyl)amino]cal L o~yl]-4-methyl-3-oxo- 1 H- 1,4-benzodiazepine-2-acetic acid Following the procedure in Example 18(c), methyl (S)-2,3,4,5-tetrahydro-7-~ 35 [[N-[(bell~i.. id~7ol-2-yl)methyl-N-(2-cyanomethyl)amino]carbonyl]-4-methyl-3-- oxo-lH-1,4-benzodiazepine-2-acetate was saponified, and the product was y~ ~;.lli7e~l from EtOH, to give the title compound as a white solid (0.420 g, W O 97/24119 PCTrUS96/20748 89%): 'H NMR ( 400 MHz, DMSO-d6) ~i 2.66 (dd, J = 16.4, 3.5 Hz, lH), 2.78 (dd, J = 16.4, 3.5 Hz, lH~, 2.92 (s, 2H), 3.80 (d, J = 16 Hz, lH), 4.52 (s, 2H), 4.84 (d, J = 2.9 Hz, 2H), 5.15 (m, lH), 5.48 (d, J = 16 Hz, lH), 6.40 (d, J = 3.5 Hz, lH), 6.54 (d, J = 8.3 Hz, lH), 7.25 (m, 4H), 7.50 (m, lH), 7.62 (m, lH); MS (ES)m/e 465 (M+H)~. Anal. Calcd for C23H22N6O4 ~ 2 HCl: C, 53.19; EI, 4.66; N, 16.18;
Found: C, 52.98; H, 4.43; N, 16.53.
Fx~ ?le 20 Preparation of (S)-2.3.4.5-tetrahydro-7-rr~ben~ ol-2-yl)methyl~methyiamino~carbonyll-3-oxo-lH-l~4-benzo~ n~-2-acetic açid a) Dimethyl (R)-2-trifluoromethanesulfonylsuccinate To a stirred, cooled (0~C) mixture of dimethyl D-malate (5.5 g, 33.9213 mmol), and dry pyridine (2.82 g, 35.7164 mmol) in dry CH2Cl2 (55 mL) was added trifluoro..,t~lh~neslllfionic anhydride (10.0 g, 35.7164 mmol) dropwise. After stirring at 0~C for 4 hr, the ~ ule was cluenched with H~O and the layers were separated.The organic layer was washed sequentially with dil HCI and brine, dried over MgSO4, and concentrated to give title compound (8.50 g, 96%) as a white solid: 'H
NMR( 250 MHz, CDCI3: ~ 3.10 (d, J = 5.8 Hz, 2H), 3.74 (s, 3H), 3.78 (s, 3H), 5.52 (t, J = 5.8 Hz, lH).
b) Dimethyl D-(2-cyanophenyl)malate A mixture of 2-arninobenzonitrile (0.5 g, 4.2323 mmol), 2,6-di-tert-bu~ylpylidine (0.85 g, 4.4439 mmol), and dimethyl (R)-2-trifluorometh~n~slllfonylsuccinate in 2:1 hexane/chloroform (25 mL) was stirred at RT for 76 hr. The mixture was concentrated, and the residue was taken up in H2O
and extracted with EtOAc. The organic extracts were washed sequentially with 10%HCI and brine, dried over MgSO4, concentrated, and purified by silica gel flash column chromatography (10% EtOAc/hexane) to give the title compound (0.886 g, 80%) as a yellow solid: 'H NMR (250 MHz, CDCl3) ~ 2.95 (d, J = 5.8 Hz, lH), 3.74(s, 3H), 3.78 (s, 3H), 4.60 (m, lH), 5.28 (d, J = 5.8 Hz, lH), 6.73 (d, J = 8.5 Hz, lH), 6.80 (t, J = 8.5 Hz, lH), 7.47 (m, 2H).
c) Methyl (S)-2,3,4,5-tetrahydro-3-oxo-lH-1,4-benzodiazepine-2-acetate A solution of dimethyl D-(2-cyanophenyl)malate (10.75 g, 41.0006 mmol) in MeOH (pre-saturated with NH3 (g) for 10 min, 100 mL) was hydrogenated over CA 0224l633 l998-06-26 W O 97/24119 PCTrUS96/20748 Ra/Ni at 55 psi for 48 hr. The catalyst was filtered off, and the filtrate was concentrated and purified by silica gel flash colurnn chromatography (40%
EtOAc/hexane) to give the title compound (5.03 g, 53%) as an off white solid: 'HNMR (250 MHz, CDCl3) o 2.65 (dd, J = 16.3, 7.6 Hz, lH), 2.99 (dd, 16.3, 5.9 Hz, lH), 3.74 (s, 3H), 3.95 (dd, J = 16, 6.9 Hz, lH), 4.79 (m, lH), 4.95 (dd, J = 16, 5.3 Hz, lH), 6.55 (t, J = 5.3 Hz, lH), 6.65 (d, J = 7.6 Hz, lH), 6.78 (d, J = 7.6 Hz, lH), 6.97(d,J=7.6Hz, lH),7.15(d,J=7.6Hz, lH).
d) Methyl (S)-2,3 ,4,5-tetrahydro-7-bromo-3-oxo- 1 H- 1 ,4-benzodiazepine-2-acetate A mixture o~ methyl (S)-2,3,4,5-tetrahydro-3-oxo-lH-1,4-benzodiazepine-2-acetate (5.03 g, 21.4746 mmol) and n-Bu4NBr3 (10.35 g, 21.4746 mmol) in CHC13 (100 mL) was stirred at RT for 3 hr, then the ~ lule was concentrated. The residue was taken up in H20, stirred, and filtered to afford the title compound (5.61 g, 83~o) as an off white solid: 'H NMR (250 MHz, CDCl3) o 2.74 (dd, J = 16.3, 7.6 Hz, lH), 3.05 (dd, J = 16.3, 5.9 Hz, lH), 3.75 (s, 3H), 4.05 (dd, J = 16, 6.9 Hz, lH), 4.73 (t, J
=5.9Hz, lH),4.86(dd,J= 16,5.3Hz, lH),6.68(d,J=7.6Hz, lH),6.75 (t,J=5.3 Hz, lH), 7.14 (s, lH), 7.25 (d, J = 7.6 Hz, lH).
e) Methyl (S)-2,3,4,5-tetrahydro-7-[[N-[(ben7imi~1~71 1-2-yl)methyl]-N-methylamino]carbonyl]-3-oxo- 1 H- 1 ,4-benzodiazepine-2-acetate A mixture of methyl (S)-2,3,4,5-tetrahydro-7-bromo-3-oxo-lH-1,4-benzodiazepine-2-acetate (1.5 g, 4.77905 mmol), 2-(methylaminomethyl)ben~imi-1~7Qle dihydrochloride (2.24 g, 9.5809 mrnol), triphenylphosphine (1.26 g, 4.7905 mmol), n-Bu3N ( 6.21 g, 33.5333 mmol), and (Ph3P)4Pd (1.10 g, 0.9581 mmol) in N-methyl 2-pyrrolidinone (20 rnL) was flushedwith argon and carbon monoxide for 10 min. The mixture was then heated at 100-105~C under a carbon monoxide balloon for 8 hr. The mixture was cooled and acidified with 6 N HCl to pH = 2. The solution was extracted with EtOAc, and theEtOAc layer was discarded. The aqueous layer was neutralized with 30% NaOH
and extracted with CH2Cl2. The organic extracts were dried over MgSO4, concentrated, and purifled by silica gel flash column chromatography (5%
MeOH/CH2Cl2) to give the title compound (1.62 g, 80%) as an off white solid: 'H
NMR (250 MHz, DMSO-d6): ~; 2.65 (dd, J = 16.3, 7.6 Hz, lH), 2.81 (dd, J = 16.3, 5.9 Hz, lH), 3.05 (s, 3H), 3.60(s, 3H), 3.75 (dd, J = 16.3, 6.9 Hz, lH), 4.78 (s, 2H), 4.95 (m, lH), 5.05 (dd, J = 16, 5.3 Hz, lH), 6.20 (d, J = 5.9 Hz, lH), 6.55 (d, J = 7.6 Hz, lH), 7.25 (m, 4H), 7.55 (m, 2H), 8.21 (t, J = 5.3 Hz, lH).
W O 97/24119 PCT~US96/20748 f) (S)-2,3,4,5-Tetrahydro-7-[[[berl7imi(1:l7Ol-2-yl)methyl]methylamino]ccubollyl]-3-oxo- 1 H- I ,4-ben70~ 7epine-2-acetic acid Following the procedure of Example 18(c), methyl (S)-2,3,4,5-tetrahydro-7-[~N-t(ben7.imi(1~7.ol-2-yl)methyl]-N-methylamino]carbonyl]-3-oxo-lH-1,4-benzodiazepine-2-acetate was saponified to afford the title compound (0.060 g, 57%) as an off white solid: 'H NMR ( 400 MHz, DMSO-d6) ~ 2.52 (dd, J = 16.3, 7.6Hz, lH), 2.84 (dd, J = 16.3, 5.9 Hz, lH), 3.20 (s, 3H), 3.75 (dd, J = 16.3, 6.9 Hz, lH), 4.95 (t, J = 5.9 Hz, lH), 5.05 (dd~ J = 16, 5.3 Hz, lH), 5.10 (s, 2H), 6.59 (d, J =
7.6 Hz, lH), 7.12 (s, lH), 7.20 (d, J - 7.6 Hz, lH), 7.48 (m, 2H), 7.69 (m, 2H), 7.90 (d, J = 5.3 Hz, lH); IR ( KBr) 3600-3100, 3100-2800, 1681, 1613, 1601, 1485, 1445, 1314, 830, 764, 742 cm l; MS (ES) m/e 422 (M+H)~. Anal. Calcd for C2,H21N5O4: C, 61.91; H, 5.20; N, 17.19. Found: C, 61.57; H, 5.32; N, 17.29.
Fx~ml?le 21 Preparation of (S~-2.3.4.5-tetrahydro-7-rrrrl-(2-hydroxyethyl)ben7imi~1~7ol-2-yllmetllyllaminolcarbonyll4-methyl-3-oxo- lH- 1.4-benzodi~epine-2-acetic açid a) Ethyl 2-[[(ben7imi-1~7O1-2-yl)methyl]amino]acetate A mixture of 2-aminomethyll~e~ 701e dihydrochloride hydrate (4.0 g, 18.1736 mmol), NaHCO3 (7.63 g~ 90.868 mmol), and ethyl 2-bromoacetate (4.55 g, 27.2603 mmol) in dry DMF (60 rnL) was stirred at RT for 24 hr, then was concentrated. The residue was taken up in H~O and extracted with CH2CI2. The organic extracts were dried over MgSO4, concentrated, and purified by silica gelflLash column chromatography (5% MeOH/CH2Cl2) to give the title compound (0.50 g, 12%) as a brown oil: 'H NMR (250 MHz, CDCI3) ~ 1.95 (s, 3H), 3.48 (s, 2H), 4.50 (m, 4H), 7.25 (m, 2H), 7.35 (m, lH), 7.73 (m, lH).
b) Methyl (S)-2,3,4,5-tetrahydro-7-[[~[1 -(2-acetyloxyethyl)ben7imi~1~7ol-2-yl]methyl]amino]~l,onyl]4-methyl-3-oxo-lH-1,4-benzodiazepine-2-acetate Following the procedure of Example 18(b), except substituting ethyl 2-[[(benzimidazol-2-yl)methyl]amino]acetate for the 4-[(ben7imid~7ol-2-yl)methyl]aminobutyronitrile, the title compound (0.251 g, 78.5%) was prepared as a white solid: ;H NMR (250 MHz, CDCl3) ~ 1.95 (s, 3H~, 2.66 (dd, J = 16.4,3.5 Hz, lH), 2.95 (s, 3H), 3.05 (dd, J = 16.4, 3.5 Hz, lH), 3.60 (d, J = 16 Hz, lH), 3.75 (s, 3H), 4.45(d, J = 5.9 Hz, 2H), 4.62 (s, 2H), 4.92 (t, J = 5.9 Hz, 2H), 5.10 ( m, lH), .~
W O 97/24119 PCTrUS96/20748 5.40(d,J= 16Hz, lH),6.49(d,J=8.3Hz, lH),7.32(m,3H),7.60(m, 2H), 7.71 (m, lH), 8.15 (t, J = 5.3 Hz, lH).
c) (S)-2.3,4,5-Tetrahydro-7-[[[[1-(2-hydroxyethyl)bçn7imi-1~7ol-2-yl]methyl]amino]carbonyl]-4-methyl-3-oxo- 1 H- 1,4-benzodiazepine-2-acetic acid To a stirred, partial suspension of methyl (S)-2.3,4,5-tetrahydro-7-[[[[1-(2-acetyloxyethyl)ben7imi~ 7ol-2-yl]methyl]amino]carbonyl]-4-methyl-3-oxo-lH-1,4-benzodiazepine-2-acetate (0.241 g, 0.475 mmol) in THF (5 mL) was added 1.0 N
LiOH (1.4 mL, 1.425 mmol). After stirring at RT overnight, the mixture was concentrated. The residue was taken up in H2O and acidified with AcOH to pH = 4.The off-white solid was filtered and lliluldt~d with acetone to afford the titlecompound (0.16 g, 75%) as a white solid: 'H NMR (400 MHz, DMSO-d6):o 2.54 (dd, J = 16.4, 3.5 Hz, lH), 2.95 (s, 3H), 3.05 (dd, J = 16.4, 3.5 Hz, lH), 3.60 (d, J =
16 Hz, lH), 4.45 (d, J = 5.9 Hz, 2H), 4.62 (s, 2H), 4.92 (t, J = 5.9 Hz, 2H), 5.10 (m, lH),5.40(d,J= 16Hz, lH),6.49(d,J=8.3Hz, lH),7.32(m,3H),7.60(m,2H), 7.71 (m, lH), 8.15 (t, J = 5.3 Hz, lH); MS (ES) m/e 452 (M+H)+. Anal. Calcd for C23H25N5O5 - 0.75 H2O: C, 59.71; H, 5.72; N, 15.14. Found: C, 59.65, H, 5.70; N,14.88.
Exarnple 22 Preparation of (+)-2.3~4.5-tetrahydro-7-rrN-(b~n7imicl~7ol-2-yl)methyl-N-rr4-(2-carboxybenzoyl)amino~butyllaminolcarbonyll-3-oxo-4-(2-phenylethyl)- 1 H- 1.4-benzot1i~7~-pine-2-acetic acid a) 4-~[(Benzimidazol-2-yl)methyllamino]butylphthz~limide A mixture of 2-aminomethylbenzimidazole dihydrochloride hydrate (22.10 g, 100.7269 mmol), NaHCO3 (42.40 g, 503.6347 mmol), and 4-bromobutylphth~limicle (34.10 g, 120.8723 mmol) in dry DMF (250 mL) was heated at 100-110~C for 6 hr, then was cooled and concenllatt;d. The residue was taken up in H2O and extracted with CH2Cl2. The organic extracts were dried over MgSO4, concentrated, and purified by silica gel flash column chromatography (5%
MeOH/CH2Cl2) to give the title compound (10.8 g, 31%) as a brown foam: IH NMR
(250 MHz, CDCl3) ~ 1.65 (m, 2H), 1.85 (m, 2H), 2.75 (t, J = 8.9 Hz, 2H), 3.78 (t, J =
8.9 Hz, 2H), 4.17 (s, 2H), 7.20 (m, 2H), 7.60 (m, 2H), 7.72 (m, 2H), 7.88 (m, 2H).
-W O 97/24119 PCT~US96/20748 b) Methyl ~+)-2,3,4,5-tetrahydro-7-[[N-(ben~i.l.if1z-7Ql-2-yl)methyl-N-[~4-(phth~limido)butyl]amino]carbonyl]-3-oxo-4-(2-phenylethyl)-lH-1,4-benzodiazepine-2-acetate To a stirred mixture of 4-[[(~çn7imi~l~7Ql-2-yl3methyl]amino]butylphth:~1imi~ (1.75 g, 5.0525 mmol), methyl (+)-2,3,4,5-tetrahydro-7-carboxy-3-oxo-4-(2-phenylethyl)- 1 H- 1,4-benzodiazepine-2-acetate (1.61 g, 4.2104 mmol), HOBt H2O (0.69 g, 5.0525 mmol), and i-PrlNEt (1.10 g, 8.4209 mmol) in dry CH3CN (30 mL) was added EDC (1.50 g, 5.0525 mmol). After stirring at RT for 24 hr, the mixture was concentrated. The residue was taken up in H2O and extracted with CH2Cl2. The organic extracts were washed sequentially with saturated NaHCO3 and brine, dried over MgSO4, concentrated, and purified by silica gel ~lash column chromatography (5% MeOH/CH2Cl2) to give the title compound (2.85 g, 95%) as a yellow foam: 'H NM~ (250 MHz, DMSO-d6) o 1.60 (m, 2H), 2.65 (m, 2H), 2.85 (dd, J = 16.4, 3.5 Hz, lH), 3.55 (m, 4H), 3.65 (s, 3H), 4.00 (d, J=
16.0 Hz, lH), 4.18 (q, J = 8.9 Hz, 2H), 4.75 (s, 2H), 5.15 (m, lH), 5.45 (d, J = 16.0 Hz, lH), 6.23 (d, J = 5.3 Hz, lH), 6.57 (d, J = 7.6 Hz, lH), 7.20 (m, 7H), 7.55 (m, 4H), 7.90 (m, 4H).
c) Methyl (+)-tetrahydro-7-[[N-[ber-~mi~1~7Ol-2-yl)methyl]-N-~4-aminobutyl)amino]carbonyl~-3-oxo~(2-phenylethyl)- 1 H- 1,4-benzodiazepine-2-acetate A mixture of methyl (+)-2~3~4~5-tetrahydro-7-[[N-(ben7imi~ 7Ql-2-yl)methyl-N-[[4-(phth~limido)butyl]amino]carbonyl]-3-oxo-4-(2-phenylethyl)-lH-1,4-benzodiazepine-2-acetate (0 50 g, 0.7015 mmol) and hydrazine (0.07 g, 2.1045mmol) in MeOH (5 mL) was refluxed for 6 hr, then was cooled and concentrated.
The residue was taken up in H2O, acidified to pH = 2 with 6N HCl, and filtered to remove a white solid. This solid was discarded. The aqueous filtrate was extracted with EtOAc, and the EtOAc layer was discarded. The aqueous layer was basified topH = 9 with Na2CO3 and extracted with CHCl3. The organic layer was dried over MgSO~ and concentrated to give the title compound (0.41 g, 89%) as an off white solid: 'H NMR (250 MHz, DMSO-d6) ~ 1.47 (m, 2H), 1.75 (m, 2H), 2.65 (m, SH), 2.85 (dd, J = 16.3, 5.9 Hz, lH~, 3.65 (s, 7H), 4.05 (d, J = 16.0 Hz, lH), 5.05 (s, 2H), 5.15 (m, lH), 5.45 (d, J = 16, lH), 6.65 (d, J = 7.6 Hz, lH), 7.25 (m, 6H), 7.41 (s, lH), 7.60 ~m, 2H), 7.85 (m, 2H).
WO 97124119 rCT/US96/20748 d) (+)-2,3,4,5-Tetrahydro-7-[[N-(ben~i...icl~7Ql-2-yl)methyl-N-[[4-(2-carboxybenzoyl)amino]butyl]amino]carbonyl]-3-oxo-4-(2-phenylethyl)- 1 H- 1,4-benzodiazepine-2-acetic acid To a stirred solution of methyl (+)-tetrahydro-7-[[N-[ben7imi-1~7Ol-2-yl)methyl]-N-(4-aminobutyl)amino]carbonyl]-3-oxo-4-(2-phenylethyl)- 1 H- 1,4-benzodiazepine-2-acetate (0.34 g, 0.47 mmol) in THF was added 1.0 N LiOH (1.2 mL). After stirring at RT overnight, the mixture was concentrated, and the residue was acidified with AcOH to pH = 4. The solid was filtered and lliLuldt~d with acetone/ether to give the title compound (0.130 g, 39%) as a white solid: 'H
NMR(400 MHz, DMSO-d6) ~ 1.47 (m, 2H), 1.75 (m, 2H), 2.54 (dd, J = 16.3, 3.5 Hz, lH), 2.65 (m, 2H), 2.85 (dd, J = 16.3, 5.9 Hz, lH~, 3.20 (m, 2H), 3.75 (m, 4H), 4.05 (d, J = 16.0 Hz, lH), 5.05 (s, 2H), 5.15 (m, lH), 5.45 (d, J = 16, lH), 6.65 (d, J = 7.6 Hz, lH), 7.25 (m, 6H), 7.41 (s, lH), 7.60 (m, 2H), 7.85 (m, 2H); IR (KBr) 3400, 3326, 3100-3000, 1721, 1637, 1626, 1616, 1607,1300, 750, 694 cm~'; MS (ES) m/e 717 (M+H)~. Anal. Calcd for C40H38N6O8 - 3 H2O: C, 63.56; H, 5.87; N, 11.12.
Found: C, 63.56; H, 5.83; N, 11.04.
Fx~ml?le 23 Preparation of (+)-2.3.4.5-tetrahydro-7-rrN-(ben7imid~7ol-2-yl)methyl~-N-rr4-(4-azido-2-hydroxybenzoyl)aminolcarbonyll -3-oxo-4-(2-phenylethyl)- 1 H- 1.4-bell7odiazepine-2-acetic acid a) Methyl (+)-2,3,4,5-tetrahydro-7-~[N-(be~7imi~1~7Ol-2-yl)methyl]-N-[[4-(4-azido-2-hydroxybenzoyl)amino~carbonyl]-3-oxo-4-(2-phenylethyl)- 1 H- 1,4-benzodiazepine-2-acetate To a mixture of methyl (+)-2~3~4~s-tetrahydro-7-[[N-(brn7imi~l~7ol-2-yl)methyl-N-[[4-(phth~limido)butyl]amino]carbonyl]-3-oxo-4-(2-phenylethyl)-lH-1,4-benzodiazepine-2-acetate (0.409 g, 0.702 mmol), 4-~.ido.c~licylic acid-N-hydroxysuccinimide ester (0.194 g, 0.702 mmol), and i-Pr2NEt (0.272 g, 2.106 mmol) in dry 2: 1 CH3CN/DMF (10 mL) was added EDC (0.25 g, 0.8424 mmol).
After stirring at RT for 24 hr, the mixture was concentrated. The residue was taken up in H2O, stirred, and ~lltered to afford, after drying, the title compound (0.204 g, 39%) as an off white solid: 'H NMR (250 MHz, DM~O-d6) ~ 1.40 (m, 2H), 1.75 (m, 2H), 2.65 (m, 3H), 2.75 (dd, J = 16.3, 5.9 Hz, lH), 3.20 (m, 2H), 3.51 (m, 4H), 3.65 (m, 3H), 3.95 (d, J = 16 Hz, lH), 4.79 (s, 2H), 5.12 (m, lH), 5.37 (d, J = 16 Hz, lH), W O 97/24119 PCTrUS96/20748 6.20 (s, lH), 6.55 (m, 3H), 7.20 (m, 8H), 7.55 (m, 3H), 7.~5 (d, J = 7.6 Hz, lH), 7.98 (s, lH), 8.75 (s, lH).
b) (+)-2,3,4,5-Tetrahydro-7-[[N-(be~ 7nl-2-yl)methyl3-N-[[4-(4-azido-2-hydroxybenzoyl)amino]carbonyl]-3-oxo4-(2-phenylethyl)- 1 H- 1,4-benzodiazepine-2-acetic acid Following the procedure of Example 22(d), methyl (+)-2,3,4,5-tetrahydro-7-[[N-(benzimidazol-2-yl)methyl]-N-[[4-(4-azido-2-hydroxybenzoyl~arninoJcarbonyl]-3-oxo-4-(2-phenylethyl)-lH-1,4-benzodiazepine-2-acetate was saponified to affordthe title compound (0.100 g, 50%) as a white solid: 'H NMR (400 MHz, DMSO-d6) ~ 1.40 (m, 2H), 1.75 (m, 2H), 2.65 (m, 3H), 2.75 (dd, J = 16.3, 5.9 Hz, lH), 3.20 (m, 2H), 3.51 (m, 4H), 3.95 (d, J = 16 Hz, IH), 4.79 (s, 2H), 5.12 (m, lH), 5.37 (d, J =
16 Hz, lH), 6.20 (s, lH), 6.55 (m, 3H), 7.20 (m, 8H), 7.55 (m, 3H), 7.85 (d, J = 7.6 Hz, lH), 7.98 (s, lH), 8.75 (s, lH); MS (ES) m/e 730 (M~H)+. Anal. Calcd for C39H39N9O6 ~ 2.~ ~20: C, 60.46; H, 5.72; N, 16.27. Found: C, 60.46; H, 5.43; N, 15.90.
F~ nple 24 rl~palalion of 2.3.4.5-tetrahydro-7-rrN-r(ben7imi(1~7ol-~-yl)methyl-N-rrr(+)-biotinoyllaminolbutyll~minolcarbonyll-3-oxo-4-(2-phenylethyl)-lH-1.4-benzo~ pine-(2RS)-acetic acid a) Methyl 2,3,4,5-tetrahydro-7-[~N-[(benzimidazol-2-yl)methyl-N-[[[(+)-biotinoyl]amino]butyl]amino~carbonyl~-3-oxo-4-(2-phenylethyl)- l H- 1,4-benzodiazepine-(2RS)-acetate To a mixture of methyl (+)-tetrahydro-7-[[N-[ben7imirl~7ol-2-yl)methyl]-N-(~aminobutyl)arnino~carbonyl]-3-oxo-4-(2-phenylethyl)- 1 H- 1,4-benzodiazepine-2-acetate (0.40 g, 0.6865 mmol), (+)-biotin (0.17 g, 0.6865 mmol), HOBt ~ H2O (0.11 g, 0.8239 mmol), and i-Pr2NEt (0.18 g, 1.3730 mmol) in 1:2 DMF/CH3CN (12 rnL) was added EDC (0.25 g, 0.8239 mmol). After stirring at RT for 24 hr, the mixturewas concentrated. The residue was taken up in H2O and extracted with CHCl3. The organic extracts were washed sequentially with saturated NaHCO3 and brine, driedover MgSO4, concentrated, and purified by silica gel flash column chromatography(10% MeOH/CH2CI2~ to give the title compound (0.24 g, 44%) as a yellow foam: lH
NMR (250 MHz, DMSO-d6) o 1.30 (m, 2H), 1.60 (m, 4H), 2.05 (t, J = 8.9 Hz, 2H);,2.60 (m, 3H), 2.68 (dd, J = 16.3, 5.9 Hz, lH), 3.10 (m, 4H), 3.45 (m, 2H), 3.60 CA 0224l633 l998-06-26 W O 97/24119 PCT~US96/20748 (m, 2H), 3.65 (s, 3H), 4.01 (d, J = 16 Hz, lH), 4.12 (t, J = 8.9 Hz, lH), 4.30 (t, J = 8.9 Hz, lH), 4.78 (s, 2H), 5.10 (m, lH), 5.45 (d, J = 16 Hz, lH), 6.20 (d, J = 5.3 Hz, lH), 6.40 (d, J = 8.9 Hz, 2H), 6.55 (d, J = 7.6 Hz, lH), 7.25 (m, 9H), 7.45 (m, lH), 7.55 (m, lH), 7.70 (t, J = 8.6 Hz, lH).
b) 2,3,4,5-Tetrahydro-7-[[N-[(bçn7imicl~7ol-2-yl)methyl-N-r[[(+)-biotinoyl]amino]butyl]amino]carbonyl]-3-oxo-4-(2-phenylethyl)-lH-1,4-benzodiazepine-(2RS)-acetic acid To a stirred solution of methyl 2,3,4,5-tetrahydro-7-[[N-[(benzimidazol-2-yl)methyl-N-[[[(+)-biotinoyl]amino]butyl]amino]carbonyl]-3-oxo-4-(2-phenylethyl)-lH-1,4-benzodiazepine-(2RS)-acetate (0.24 g, 0.2967 mmol) in 1:2 THF/MeOH (6 mL) was added 1.0 N LiOH (0.44 mL). After stirring at RT overnight, the mixture was concentrated. The residue was taken up in H2O and acidified with AcOH to pH
= 4. The off-white solid was filtered, and triturated with hot acetone to give the title compound (0.160 g, 68%) as a white solid: 'H NMR (400 MHz, DMSO-d6) d 1.30 (m, 2H), 1.60 (m, 4H), 2.05 (t, J = 8.9 Hz, 2H), 2.60 (m, 3H), 2.68 (dd, J = 16.3, 5.9 Hz, lH), 3.10 (m, 4H), 3.45 (m, 2H), 3.60 (m, 2H), 4.01 (d, J = 16 Hz, lH), 4.12 (t, J
= 8.9 Hz, lH), 4.30 (t, J = 8.9 Hz, lH), 4.78 (s, 2H), 5.10 (m, lH), 5.45 (d, J = 16 Hz, lH), 6.20 (d, J = 5.3 Hz, lH), 6.40 (d, J = 8.9 Hz, 2H), 6.55 (d, J = 7.6 Hz, lH), 7.25 (m, 9H), 7.45 (m, lH), 7.55 (m, lH), 7.70 (t, J--8.6 Hz, lH); MS (l~S) m/e 795 (M+H)~. Anal. Calcd for C42H5"N806S 1.75 H20: C, 61.04; H, 6.52; N, 13.56.
Found: C, 60.89; H, 6.24; N, 13.31.
F.x~ml?le 25 P.epa~a~ion of (+)-2.3~4~5-tetrahydro-7-rrN-r(benzimidazol-2-yl)methyl~-N-(4-aminobutyl)~minolcarbonyll-3-oxo-4-(2-phenylethyl)-lH-1.4-benzodiazepine-2-acetic acid a) (+)-2,3,4,5-Tetrahydro-7-[[N-[(ben7imid~7ol-2-yl)methyl]-N-(4-aminobutyl)amino]carbonyl]-3-oxo-4-(2-phenylethyl)- lH- 1,4-benzodiazepine-2-acetic acid Following the procedure in Example 24(b), methyl (+)-2,3,4,5-tetrahydro-7-[[N-[ben7imifl~701-2-yl)methyl]-N-(4-aminobutyl)amino]carbonyl]-3-oxo-4-(2-phenylethyl)-lH-1,4-benzodiazepine-2-acetate was saponi~led to give the title compound (0.250 g, 80%) as an off white solid: 'H NM~ (400 MHz, DMSO-d6) 1.37 (m, 2H), 1.62 (m, 2H), 2.52 (dd, J = 3.5 Hz, lH), 2.64 (m, 2H), 2.75 (dd, J =
CA 0224l633 l998-06-26 W O 97/24119 PCT~US96/Z0748 16.3, 5.9 Hz, lH), 3.51 (m, 4H), 3.91 (d, J = 16 Hz, lH), 4.98 (s, 2H), 5.05 (m, lH),5.37(d,J= 16Hz, lH),6.53(d,J=7.9Hz,2H),7.17(m,7H),7.52(m, lH), 7.62 (s, lH), 7.78 (m, lH); IR ( KBr): 3386, 3100-3000, 1647, 1613, 1403, 740, 699 cm '; MS (ES) m/e 569 (M+H)+. Anal. Calcd for C32H36N604 ~ 2.75 H20: C, 62.18; H, 6.77; N, 13.60. Found: C, 62.11; H, 6.68; N, 13.57.
Fxzlrnple 26 Preparation of (+)-2~3.4~5-tetrahydrb-7-rfN-r(benzimidazol-2-yl)methyll-N-rr4-(4-azido-3-iodo-2-hydroxyben7oyl~aminolbutyllaminolc~1Jon~111-3-oxo-4-(2-ph~r~ylethyl)- 1 H- 1.4-ben~otli~7~pine-2-acetic acid a) 3-Iodo-4-~7i~1os~1icylic acid-N-hydroxysllccinimifl. ester To a stirred mixture of 4-azidosalicylic acid N-hydroxysuccinimide ester (0.500 g, 1.8103 mmol) and silver trifluoroacetate (0.44 g, 1.9913 mmol) in CHCI3 (10 mL) was added iodine (0.510 g, 1.9913 mmol). After stirring at RT overnight,the reaction was filtered to remove a solid precipitate. The filtrate was washedsequentially with H2O, saturated NaHCO3 and brine, then was dried over MgSO~.
Concentration gave the title compound (0.703 g, 97%) as a light purple solid: 'HNMR (250 MHz, CDCl3) ~ 2.98 (s, 4H), 6.83 (d, J = 7.6 Hz, lH), 8.05 (d, J = 7.6 Hz, lH).
b) Methyl (+)-tetrahydro-7-[[N-[(ben7imic1~7O1-2-yl)methyl]-N-[[4-(4-azido-3-iodo-2-hydroxybenzoyl)amino]butyl]arnino]carbonyl]-3-oxo-4-(2-phenylethyl)-lH-1.4-benzodiazepine-2-acetate Following the procedure in Example 22(b), except substituting 3-iodo-4-azidosalicylic acid-N-hydroxysuccinimide ester for the 4-[[(~n7imi~1~7ol-2-yl)methyl]amino~butylph~h~limicl~, the title compound (0.312 g, 56%) was prepared as a yellow foam: 'H NMR (250 MHz, DMSO-d6) ~ 1.42 (m, 2H), 1.60 (m, 2H), 2.52 (dd, J = 16.3, 3.5 Hz, lH), 2.63 (m, 2H), 2.79 (dd, J = 16.3, 5.9 Hz, lH), 3.25 (s, 2H), 3.55 (m, 6H), 3.65 (s, 3H), 3.95 ~d, J = 16 Hz, lH), 4.75 (s, 2H), 5.02 (m, lH), 5.35 (d, J = 16 Hz, lH), 6.14 (d, J = 5.3 Hz, lH), 6.52 (d, J = 7.9 Hz, lH), 6.86 (d, J = 7.9 Hz, lH~, 7.25 (m, lOH), 7.51 (s, 2H), 7.90 (d, J = 7.9 Hz, lH), 9.01 (s, lH).
W O 97/24119 PCTrUS96/20748 c) (+)-Tetrahydro-7-[[N-[(bçl ~ i . I ,i(1~7Ol-2-yl)methyl~-N-[~4-(4-a_ido-3-iodo-2-hydroxybenzoyl)amino]butyl] amino]carbonyl] -3 -oxo-4-(2-phenylethyl)- 1 H-1.4-ben7.o~ 7~pine-2-acetic acid Following the procedure in Example 22(c), methyl (+)-tetrahydro-7-[[N-[(bç~ 7Ol-2-yl)methyl]-N-[[4-(4-a7ido-3-iodo-2-hydroxybenzoyl)amino]butyl]amino]carbonyl] -3-oxo4-(2-phenylethyl)- 1 H- 1.4-- benzodiazepine-2-acetate was saponified. Purification by silica gel flash colurnn chromatography (0.5, 0.5, 9.5 AcOH/MeOH/CH2Cl2) gave the title compound (0.170 g, 58%) as an off white solid: 'H NMR (400 MHz, DMSO-d6) ~ 1.42 (m, 2H), 1.60 ~m, 2H), 2.52 (dd, J = 16.3, 3.5 Hz, lH), 2.63 (m, 2H), 2.79 (dd, J = 16.3, 5.9 Hz, lH), 3.25 (s, 2H), 3.55 (m, 6H), 3.95 (d, 16, lH), 4.75 (s, 2H), 5.02 (m, lH), 5.35 (d, J= 16Hz, lH),6.14(d,J=5.3Hz, lH),6.52(d,J=7.9Hz, lH),6.86(d,J=7.9 Hz, lH), 7.25 (m, 10H), 7.51 (s, 2H), 7.90 (d, J = 7.9 Hz, lH), 9.01 (s, lH); MS (ES) mle 856 (M+H)+; IR (KBr): 3360, 3100-3000, 2116, 1704, 1643, 1610, 1586, 1477, 1305, 1274, 766, 700 cm~'. Anal. Calcd for C39H38IN906 ~ 4.5 H20: C, 50.01; H, 5.06;
N, 13.46. Found: C, 50.19; H, 5.01; N, 13.12.
Fx~ml?le 27 Preparation of 5-rrr(ben7imid~7ol-2-yl)methyllmethylaminolcarbonyll-lH-be~ 7Ole-2-aminoacetic acid a) Methyl 5-[~[(benzimida_ol-2-yl)methyl]methylamino]carbonyl]-lH-benzimidazole-2-;~mino~cetate Diisopropylethylamine (1.1 mL, 6.48 mmol) was added to a stirred solution of methyl 5-carboxy-b~n7imi~1~7Ole-2-~mino~et~t~ (0.24 g, 0.96 mmol), 2-(methylaminomethyl)bellzil,.idazole bis-trifluoroacetate (0.56 g, 1.44 mmol), HOBt H2O (0.19 g, 1.44 mmol), and EDC (0.28 g, 1.44 mmol) in anhydrous DMF (8 mL) at RT. After 23 h, the reaction mixture was diluted with CH2Cll (100 mL) and washed sequentially with 5% NaHCO3 (30 mL) and brine (30 mL). Drying (MgSO4), concentration, and silica gel chromatography (10% MeOH/CH2Cl2) gave the title compound (0.16 g, 42%~ as an off-white solid: MS (ES) m/e 393.0 (M+H)+.
A
35 b) 5-[[[(Ben7imicl~701-2-yl)methyl]methylatnino~carbonyl]-lH-bçn7imic~7Ole-2- aminoacetic acid W O97/24119 PCT~US96~0748 1.0 N LiOH (1.0 rnL, 1.0 mmol) was added dropwise at RT to a mixture of methyl 5-[[r(ben7imid~7-~1-2-yl)methyl]methylamino]carbonyl]-lH-ben7.imi~1~701e-2-aminoacetate (0.16 g, 0.41 mmol) in THF (10 rnL) and H20 (10 mL). After 1 h, the reaction ~ Lule was concentrated to a small volume on the 5 rotavap and cooled in an ice bath before neutralizing with 1.0 N AcOH (1.0 mL).
The solid was collected, washed with cold H20, and air dried to give the title compound (0.15 g, 100%) as an off white solid: MS (ES) rn/e 379.2 (M+H)+.
~F~m,J?le 28 I~e~aldlion of (_)-2.3~4~s-Tetrakydro 7-rrrfbenzimidazol-2-yl)methyllmethy!;lmino~carbonyll-4-(3.3-~lim~thylbutyl)-3-oxo-lH-1.4-ben7n~ 7epine-2-acetic acid a) Methyl (V-2,3,4,5-tetrahydro 7-[[[(bc~7imi-1~7ol-2-yl)methyl]methylarnino]carbonyl]-4-(3,3-dimethylbutyl)-3-oxo- lH- 1,4-benzodiazepine-2-acetate Diisopropylethylamine (0.94 rnL, 5.4 mmol) was added to a stirred solution of methyl (_)-7-carboxy-4-(3,3 -dimethylbutyl)-3-oxo-2,3,4,5-tetrahydro- 1 H- l ,4-benzodia_epine-2-acetic acid (0.39 g, 1.08 mmol), 2-(methylamin~lllet}lyl)ben7imi~1~701e bis(trifluoroacetate) (0.42 g, 1.08 rnmol), ), EIOBt H2O (0.22 g, 1.62 mmol), and EDC (0.31 g, 1.62 mmol) in anhydrous DMF
(8 mL) at RT. After 23 h, the reaction mlixture was diluted with CH2Cl~ (100 mL)and washed sequentially with 5% NaHCO3 (2 x 25 ml) and brine (25 mL). Drying (MgSO4), concentration, and silica gel chromatography (7% MeOH/CH2Cl2) gave the title compound (0.39 g, 71%) as a white solid: MS (ES) m/e 506.4 (M+H)+.
b) (i)-2,3,4,5-Tetrahydro 7-~[[(bç~ 7Ol-2-yl)methyl]methylamino]carbonyl]-4-(3,3-dimethylbutyl) -3 -oxo- 1 H- 1,4-benzodia_epine-2-acetic acid 1.0 N LiOH (1.Q rnL, 1.0 mrnol) was added dropwise at RT to a mixture of methyl (i)-2,3,4,5-tetrahydro 7-~[(b~imi~7(~l-2-yI)methyl]methylamino]carbonyl]-4-(3,3-dimethylbutyl)-3-oxo- lH- 1,4-benzodiazepine-2-acetate (0.38 g, 0.75 mmol) in THF (10 mL) and H2O (10 mL).
After 50 min, the reaction mixture was concentrated to a small volume on the rotavap and cooled in an ice bath before neutralizing with 1.0 N AcOH (2.5 mL).
The solid was collected, washed with a cold H20, and air dried to give the titlecompound (0.27 g, 73%) as a white solid: MS (ES) rn/e 492.2 (M~H)+.
W O 97124119 PCTrUS96/20748 F.xample 29 Preparation (_)-2.3.4.5-Tetrahydro 7-rrr(ben7imi{~7ol-2-yl)methyl7aminolcarbonyll-4-(3 .3-dimethylbutyl)-3-oxo- 1 H- 1 .4-benzodiazepine-2-acetic acid -a) Methyl (+)-2,3,4,5-tetrahydro 7-[l[~be.n7imi~1~7~.1-2-yl)methyl]amino]carbonyl]-4-(3,3-dimethylbutyl)-3-oxo- lH-1,4-benzodiazepine-2-acetate Diisopropylethylamine (0.79 rnL, 4.56 mmol) was added to a stirred solution of methyl (+)-7-carboxy-4-(3 ,3-dimethylbutyl)-3-oxo-2,3,4,5-tetrahydro- lH- 1,4-benzodiazepine-2-acetic acid (0.33 g, 0.91 mmol), 2-(aminomethyl)ben7imi~1~7Ole dihydrochloride hydrate (0.3 g, 1.36 mmol), HOBt H2O (0.18 g, 1.36 mmol), and ED(~ (0.26 g, 1.36 mmol) in anhydrous DMF (8 mL) at RT. After 20 h, the reaction15 rnixture was diluted with CH Cl2 (70 mL) and washed sequentially with 5% NaHCO3 (2 x 20 mL) and brine (20 mL). Drying (MgSO4), concentration, and silica gel chromatography (7% MeOH/CH2Cl2) gave the title compound (0.25 g, 56%) as a white solid: MS (ES) m/e 492.4 (M+H)+.
20 b) (_)-2,3,4,5-Tetrahydro 7-[[[(be~ 7Ol-2-yl)methyllamino]carbonyl]4-(3,3-dimethylbutyl)-3-oxo-lH-1,4-benzotli:l7epin~-2-acetic acid 1.0 N LiOH (1.0 mL, 1.0 mmol) was added dropwise at RT to a mixture of methyl ( )-2,3,4,5-tetrahydro 7-[[[(benzimidazol-2-yl)methyl~amino]carbonyl]-4-(3,3-dimethylbutyl)-3-oxo-lH-1,4-benzodiazepine-2-acetate (0.24 g, 0.49 mmol) in25 THF (8 mL) and H2O (8 mL). After 2.5 h, the reaction mixture was concentrated to a small volume on the rotavap and cooled in an ice bath before neutralizing with 1.0 N AcOH (1.2 mL). The solid was collected, washed with cold H20 ,and air dried togive the title compound (0.25 g, 109%) as a white solid: MS (ES) m/e 478.2 (M+H)+.
l~xample 30 Plc;palation of (+)-2.3.4~5-Tetrahydro 7-rrr(4-azaben7.i~ 7ol-2-yl)methyllmethylaminolcarbonyl~-4-(3 .3-dimethylbutyl)-3-oxo- 1 H- 1.4-35 benzodiazepine-2-acetic acid CA 0224l633 l998-06-26 W O 97t24119 PCTrUS96/20748 a) Methyl (+)-2,3,4,5-tetrahydro 7-[[[(4-~7~hen7imi~l~7~ 2-yl)methyl]methylamino]carbonyl]-4-(3,3-dimethylbutyl)-3-oxo- 1 H- 1,4-benzodiazepine-2-acetate Diisopropylethylamine (0.53 mL, 3.0 mmol) was added to a stirred solution of methyl (+)-7-carboxy-4-(3,3-dimethylbutyl)-3-oxo-2,3,4,5-tetrahydro- IH- 1,4-benzodiazepine-2-acetic acid (0.22 g, 0.61 mmol), 2-(methylamino)methyl-4-azaben7imi~1~701e diacetate (0.29 g, 1.0 mmol), ~IOBt ~ H2O (0.12 g, 0.91 mmol),and EDC (0.17 g, 0.91 mmol) in anhydrous CH3CN (12 mL) at RT. After 21 h, the reaction mixture was conce~ dl~d~ diluted with CEI~Cl2 (100 mL), and washed sequentially with 5% NaHCO3 (2 x 20 mL) and brine (20 mL). Drying (MgSO4), concentration, and silica gel chromatography (7% MeOH/CH2Cl2) gave the title compound (0.147 g, 48%) as a white solid: MS (ES) m/e 507.4 (M+H)+.
b) (+)-2,3,4,5-Tetrahydro 7-[[[(4-~7~h~ 7ol-2-yl)methyl]methylamino]carbonyl]-4-(3,3-dimethylbutyl)-3-oxo-lH-1,4-benzodiazepine-2-acetic acid 1.0 N LiOH (0.69 mL, 0.69 mmol) was added dropwise at RT to a rnixture of methyl (+)-2,3,4,5-tetrahydro 7-[[[(4-azaben7i,..il1~7Ol-2-yl)methyl]methylamino]carbonyl]-4-~3,3-dimethylbutyl)-3-oxo-lH- 1,4-benzodiazepine-2-acetate (0.14 g, 0.276 mmol) in THF (8 mL) and H2O (8 mL).
After 2 h, the reaction n~ib~ ; was concentrated to a small volume on the rotavap and cooled in an ice bath before neutralizing with 1.0 N AcOH (0.69 mL). The solid was collected, washed with cold H20, and air dried to give the title compound (0.074 g, 54%) as a white solid: MS (ES) m/e 493.2 (M+H)+.
F;.x~ml?le 31 Preparation of (Sl-2~3.4.5-tetrahydro-7-rrrl l-r(ben7imicl~7Ql-2-yl~m~tl~yllben7imicl~7ol-2-yllmetllyllarnino~cal1,o,l~1l-4-methyl-3-oxo-lH-l ~4-benzodiazepine-2-acetic acid - a) Methy} (S)-2,3,4,5-tetrahydro-7-[[[[l-[(benzimid~ol-2-yl)methyl]bçn7imi~ ol-2-yl]methyl]amino~carbonyl]-4-methyl-3-oxo-lH-1,4-ben7O~ 7epine-2-acetate Diisopropylethylamine (0.27 mL, 1.53 mmol) was added to a stirred solution of methyl (S)-7-carboxy-4-methyl-3-oxo-2,3,4,5-tetrahydro- 1 H- 1,4-benzodi~epine-2-acetate trifluoroacetate (0.14 g, 0.34 mmol), 2-[[l-[(ben7imi(i~7Ol-2-yl)methyl]be~7imitl~7c~e]methyl~amine bis(trifluoroacetate) (0.17 g, 0.34 mmol), 150 W O 97/24119 PCTrUS96/20748 HOBt H2O (0.064 g, 0.48 mmol), and ~DC (0.091 g, 0.48 mmol) in anhydrous DMF (10 mL) at RT. After 22 h, the reaction mixture was concentrated, diluted with CH2Cl2 (70 mL), and washed sequentially with 5% NaHCO3 (2x30 mL) and brine (20 mL). Drying (MgSO4), concentration, and silica gel chromatography (7% MeOH/CH2Cl2) gave the title compound (0.080 g, 43%) as a white solid: MS
~ (ES) m/e 552.2 (M+H)+.
b) (S)-2,3,4,5-Tetrahydro-7-[[~rl-[(ben7imid~7-~1-2-yl)methyl]ben7imi~ 7ol-2-yl]methyl]amino]carbonyl]-4-methyl-3-oxo-lH-1,4-benzodiazepine-2-acetic acid 1.0 N LiOH (0.36 mL, 0.36 mmol) was added dropwise at RT to a rnixture of methyl (S)-2,3,4,5-tetrahydro-7-[[[[l-[(ben7i~ 7ol-2-yl)methyl]ben7imi~t~7Ql-2-yl]methyl]amino]carbonyl]-4-methyl-3-oxo- lH- 1,4-benzodiazepine-2-acetate (0.08g, 0.14 mmol) in THF (5 mL) and H2O (4 mL). After 2 h, the reaction mixture was concentrated to a small volume on the rotavap and cooled in an ice bath before neutralizing with 1.0 N AcOH (0.69 mL). The solution was lyophilized to a crude product (0.086 g) as a white powder. Purification on C-18 Bond Elute (0%-20%
CH3CN/H2O cont:~ining 0.1 % TFA) afforded the title compound as a white powder:
MS (ES) m/e 538.2 (M+H)+.
Example 32 Prep~r~tion of (S)-2.3.4.5-Tetrahydro-7-rrbisr(ben7imicl~7ol-2-yl)methyllarninolcarbonyll-4-methyl-3-oxo-lH-1.4-benzodiazepine-2-acetic acid a) Methyl (S)-2,3,4,5-tetrahydro-7-[rbis[(ben7imi~1~7ol-2-yl)methyl]amino~carbonyl]-4-methyl-3-oxo- 1 H- 1,4-benzodiazepine-2-acetate Diisopropylethylamine (0.22 mL, 1.29 mmol) was added to a stirred solution of methyl (S)-7-carboxy-4-methyl-3-oxo-2,3,4,5-tetrahydro- 1 H- 1,4-benzodiazepine-2-acetate trifluoroz-~-et~te (0.075 g, 0.26 mmol), bis[(ben7imi<1~7Ol-2-yl)methyl]amine tris(trifluoro~cet~te) (0.16 g, 0.26 mmol), HOBt ~ H2O (0.05 g, 0.36 mmol), and EDC (0.069 g, 0.36 mmol) in anhydrous CH3CN (10 mL) at RT. After 17 h, the reaction rnixture was concentldted, diluted with CH2Cl2 (80 mL), and washed sequentially with 5% NaHCO3 (2 x 20 mL) and brine (20 mL). Drying - (MgSO4), concentration, and silica gel chromatography (7% MeOH/CH2Cl2) gave the title compound (0.05 g, 36%) as a white solid: MS (ES) m/e 552.2 (M+H~+.
-:
W O 97/24119 PCT~US96/20748 b) (S)-2,3,4,5-Tetrahydro-7-[[bist(ben7irni~ 7nl-2-yl)methyl]amino]carbonyl]-4-methyl-3-oxo-lH-1,4-benzodia_epine-2-acetic acid 1.0 N LiOH (0.23 mL, 0.23 mmol) was added dropwise at RT to a mixture of methyl ~S)-2,3,4,5-tetrahydro-7-[[bis[(ben7imi~7ol-2-yl)methyl]amino]carbonyl]-4-methyl-3-oxo-lH-1,4-benzodiazepine-2-acetate (0.05 g, 0.09 mmol) in THF (6 mL) and H20 (4 rnL). After 1 h, the reaction ~l~ixAluie was concentrated to a small volume on the rotavap and cooled in an ice bath before neutralizing with 1.0 N
AcOH (0.3 mT.). The solid was collected, washed with cold H~O, and air dried to give the title compound (0.048 g, 98%) as a white solid: MS (ES) m/e 538.2 (M+H)+.
Fx~mple 33 P~pdld~ion of f_)-2.3.4.5-tetrahydro-7-rrbisr~ben7imi~7ol-2-yl)methyllamino~carbonyll-3-oxo-4-(2-phenethyl)-lH-1 ~4-benzofli~7~ 1~ine-2-acetic B~
a) Methyl (+)-2,3,4,5-tetrahydro-7-[[bis[(ben7imi<1~7Ol-2-yl)methyl]amino]carbonyl~-3-oxo-4-(2-phenethyl)-lH-1,4-berl7o~ 7Ppine-2-acetate Diisopropylethylamine (0.3 mT, 1.74 mmol) was added to a stirred solution of methyl (+)-7-carboxy-3-oxo-2-(2-phenylethyl)-2,3,4,5-tetrahydro- lH- 1,4-benzodiazepine-2-acetate (0.11 g, 0.29 rnmol), bis[(ben7imi-1~7Ol-2-yl)methyl]amine tris(trifluoroacetate) (0.18 g, 0.29 mmol), HOBt ~ H2O (0.058 g, 0.43 mmol), andEDC (0.083 g, 0.43 mmol) in anhydrous CH3CN (12 mL) at RT. After 21 h, the reaction mixture was concelltld~d, diluted with CH2Cl2 (100 rnL), and washed sequentially with 5% NaHCO3 (2 x 20 mL) and brine (20 mL). Drying (MgSO4), concentration, and silica gel chromatography (7% MeOH/CH2CI2) gave the title compound (0.13 g, 70%) as a white solid: MS (ES) m~e 642.2 (M+H)+.
b) (+)-2,3,4,5-Tetrahydro-7-[[bis[(ben7imi-1~7nl-2-yl~methyl]amino]carbonyl]-3-oxo~-(2-phenethyl)- 1 H- l ,4-benzodiazepine-2-acetic 1.0 N LiOH (0.6 mL, 0.6 rnmol) was added dropwise at RT to a mixture of methyl (~)-2~3~4~5-tetrahydro-7-[[bis~(be~7~ 7nl-2-yl)methyl]amino]carbonyl]-3 oxo-4-(2-phenethyl)- 1 H- 1,4-benzodiazepine-2-acetate (0.13 g, 0.20 mmol) in THF
(5 mL) and H20 (5 rnL). After 18 h, the reaction mixture was concentrated to a small volume on the rotavap and cooled in an ice bath before neutralizing with 1.0 N
AcOH (0.6 mT.). The solution was Iyophilized to a crude product (0.092 g, 77~o) as .
W O 97/24119 PCTrUS96/20748 a white powder. ODS chromatography (step gradient, 5%-30%
CH3CN/H2O cont~inin?~ 0.1 % TFA) afforded the title compound as a white powder:
MS (ES) mle 628.2 (M+H)+.
Ex:~ml?le 34 Preparation of ethyl (~)-3-rrr2-(ben7imidazol-2-yl~ethyl~aminolsuccinoyllamino-4-pentynoate a) (+)-4-Ethynyl-2-azetidinone 4-Acetoxy-2-~7~ti(1inone (9.0 g, 69.7 mmol) was added slowly to a solution of ethynylm~gne~iumchloride (31 mL of a 0.5 M THF solution, 0.55 mmol) at 0~C.
After 1.5 h, 1.0 N HCl (100 mL) was added, and the mixture was taken up in EtOAc(300 mL) and washed sequentially with 1.0 N HCl (100 mL), saturated NaHCO3 (100 mL), and brine (100 mT.). After drying (MgSO4) and concentration, the titlecompound (4.57 g, 69%) was obtained as a light brownish solid: MS (ES) mJe 96.0 (M+H)+.
b) Ethyl (+)-3-amino-4-pentynoate A mixture of (_)-4-ethynyl-2-:~7~ tirlinnne (1.3 g, 13.68 mmol), EtOH (54 ml), and concentrated HCl (6 mL) was heated to reflux for 18 h. The reaction wascooled to RT before adjusting the pH to 8.0 using saturated NaHCO3. The reactionwas extracted with EtOAc (3 x 70 mL), and the combined EtOAc layers were washed with brine (50 ml). Drying (MgSO4) and concentration gave the title compound (1.06 g, 55%) as a brownish liquid: MS (ES) m/e 141.9 (M+H)+.
c) Methyl-[[2-(benzimid~ol-2-yl)ethyl]amino]succinate 3-Carbomethoxypropionylchloride(0.6 g, 4.0 mmol) was added at 0~C to a stirred solution of 2-aminoethylben7imi~ 701e ~ eet~te (1.13 g, 4.0 mmol) and diisol~lu~ylethylamine (2.59 g, 20 mmol) in dry CHzClz (45 rnL). After stirring for 1.5 h at RT, the reaction mixture was diluted with CH2Cl2 (50 mr ) and washed sequentially with H2O (30 mL), 5% NaHCO3 (30 ml), and brine (30 mL). Drying (MgSO4), concentration, and silica gel chromatography (8% MeOH/CH2C12) gave the title compound (0.2 g, 18%) as a yellow solid: MS (E~) m/e 276.4 (M+H)+.
. d) [[2-(Ben7imi~1~7-)1-2-yl)ethyl]amino]succinic acid W O 97/24119 PCT~US96/20748 A mixture of methyl-[[2-(ben7imi~ 7Ol-2-yl)ethyl]annino]succinate(0.2 g, 0.73 m~nol), 1.0 N NaOH (1.82 ml, 1.82 mmol) and MeOH (10 rnL) was stirred at RT for 24 h, then was concentrated to dryness. H2O
(5 mL) was added, the solution was neutralized with 1.0 N HCl (1.82 mL), and theresnlt;ng solution was lyophilized to give the crude title compound (0.23 g) as an off-white powder: MS (ES) m/e 261.9 (M+H)+.
e) Ethyl (_)-3-[~[2-(ben7.imi-1~7~ 2-yl)ethyl]amino]succinoyl]amino-4-pentynoateDiisopropylethylamine (0.32~mL, 1.83 mrnol) was added to a stirred solution of ethyl (+)-3-amino-4-pentynoate (0.12 g, 0.88 mmol), [[2-(be~ 7Ql-2-yl)ethyl]arnino]succinic acid (0.19 g, 0.73 mmol), HOBt H20 ~0.15 g, 1.1 mmol), and EDC (0.21 g, 1.1 mmol) in anhydrous CH3CN (15 ml) and DMF (3 mL) at RT.
After 23 h, the reaction mixture was concentrated, diluted with CH2Cl2 (100 mL),and washed sequentially with 5% NaHCO3 (2 x 25 mL) and brine (25 mL). Drying (MgSO4), concentration, and silica gel chromatography (7% MeOH/CH2Cl2) gave the title compound (0.07 g, 25%) as a white solid: MS (ES) mle 385.4 (M+H)+.
F.~mRle 35 P~ ~dlion of (_1-3-rrr2-(Ben7imitl~7ol-2-yl)ethyllaminolsuccinoyllamino-4 pentynoic acid a) (+)-3-r[r2-(Ben7imifi~7- 1-2-yl)ethyl]amino]succinoyl]amino-4-pentynoic acid 1.0 N LiOH (0.78 mL, 0.78 rnmol) was added dropwise at RT to a rnixture of ethyl (+)-3-[r[2-(bçn7imi~ 7ol-2-yl)ethyl]amino]succinoyl]amino-4-pentynoate (0.12 g, 0.31 mmol) in THF (5 mL), H2O (5 ml) and CH3CN (1 mL). After 3 h, the reaction mixture was concentrated to a small volume on the rotavap and cooled in an ice bath before neutralizing with 1.0 N AcOH (0.78 mL). The solution was lyophili7~l1 to a crude product (0.167 g) as a white powder. ODS chromatography (10% CH3CN/H2O cont~lining 0.1% TFA) afforded the title compound as a white powder: MS (ES) m/e 357.1 (M+H)+.
~;xample 36 Prep:lration of (+)-3-rrr4-(4-Azal~el.7i.. id~zol-2-yl)butanoyll~lycyllaminol-4-pentynoic acid (SB-237554) W O 97/24119 PCTrUS96/20748 a) Methyl (4-azabenzimidazol-2-yl)butyrate Triethylamine (319 mL, 22.9 mmol) was added to a mixture of 2,3-diaminopyridine (2.5 g, 22.9 mmol) and methyl 4-(chloroformyl)butyrate (3.77g, 22.9 mmol) in dry THF (50 mL) at 0~C. After stirring for 16 h at RT, the reaction was concentrated to dryness under vacuum. The residue was dissolved in glacial AcOH (25 mL) and was heated at 110~C. After 93 h, the reaction was allowed to - cool to RT and was conct;ntldL~;d under vacuum. The dark brown residue was diluted with H2O (40 mL) and CH2Cl2 (40 mL), and the mixture was neutralized to pH 7 using 5 N NaOH. The layers were separated and the aqueous layer was furtherextracted with CH2C12 (2x100 mL). The combined organic layers were washed sequentially with 5% NaHCO3 (2 x 30 mL)and brine (30 mL). Drying (MgSO4), concentration, and silica gel chromatography (7% MeOH/CH2Cl2) gave the title compound (0.47, 9%): MS (ES) m/e 220.0 (M+H)+.
~5 b) (4-Azabçn7imi(1zl7Ql-2-yl)butyric acid A lllii'~Ul'~ of methyl (4-:~7zlhen7imidazol-2-yl)butyrate (0.47 g, 2.13 mmol), 1.0 N NaOH (6 mL, 6.0 mmol) and MeOH (10 mL) was stirred at RT for 5.5 h, then was concentrated to dryness. The residue was diluted with H2O (2 mL) and neutralized with 1.0 N HCl (0.73 mL). The resulting solid was collected and air dried to give the title compound (0.32 g, 73%) as a yellow powder: MS (ES) m/e 206.0 (M+H)+.
c) Ethyl (_3-3-[[(N-tert-butoxycarbonyl)glycyl]amino]-4-pentanoate Diisopropylethylamine (0.92 mL, 5.32 mmol) was added to a stirred solution of ethyl I_)-3-amino-4-pentynoate (0.3 g, 2.13 mmol), Boc-Gly (0.56 g, 3.19 mmol), HOBt H2O (0.43 g, 3.19 mmol), and EDC (0.61 g, 3.19 mmol) in anhydrous CH3CN (15 mL) at RT. After 34 h, the reaction ll~Ult; was concentrated, diluted with CH2Cl2 (70 mL), and washed sequentially with 5% NaHCO3 (2x15 mL) and brine (15 mL). Drying (MgSO4), concentration, and silica gel chromatography (1:1EtOAc/Hexane) gave the title compound (0.5 g, 79%) as a colorless oil: MS (ES) m/e 299.2 (M+H)+.
d) Ethyl (:!~)-3-~(glycyl)amino}-4-pPnt~no~tl~ trifluoroacetate A solution of TFA (5 mL) and CH2Cl2 (15 mL) at RT was added all at once to ethyl (_)-3-~[(N-tert-butoxycarbonyl)glycyl]amino]-4-pe~t~no~te (0.5 g, 1.68 mmol). After 30 min, the solution was concentrated on the rotavap, and the residue W O 97/24119 PCTrUS96/20748 was reconcentrated from toluene (to remove residual TFA) to afford the title compound (0.55g, 106%) as a light yellow syrup: MS (ES) m/e 199.2 (M+H)+.
e) Ethyl (+)-3-~[[4-(4-azaben7imi~ 2-yl)butanoyl]glycyl]amino}-4-pentynoate Diisopropylethylamine (0.94 mL, 5.43 mmol) was added to a stirred solution of ethyl (+)-3-[(glycyl)amino]-4-pçnt~n-)~tlq trifluoro~l~et~te (0.55 g, 1.76 mmol), (4-azabenzimidazol-2-yl)butyric acid (0.32 g, 1.55 mmol), HOBt ~ H2O (0.31 g, 2.33 mmol), and EDC (0.45 g, 2.33 mmol) in anhydrous CH3CN (15 mL) at RT. After 64 h, the reaction mixture was concentrated, diluted with CH2Cl2 (100 mL), and washed sequentially with 5% NaHCO3 (2 x 25 mL) and brine (25 rnL). Drying (MgSO4), concentration, and silica gel chromatography (7% MeOH/CH2CI2) gave the title compound (0.11 g, 18%) as a white solid: MS (ES) m/e 386.4 (M+H)+.
f) (_)-3-[~[4-(4-Azaben7imi~1~7ol-2-yl)butanoyl]glycyl]amino]-4-pentynoic acid 1.0 N LiOH (0.71 mL, 0.71 mmol) was added dropwise at RT to a mixture of ethyl (+)-3-[[[4-(4-~abt n7imi~ 7ol-2-yl)butanoyl]glycyl~amino]-4-pentynoate (0.11 g, 0.285 mmol) in THF (5 mL), H2O (5 mL) and CH3CN (1 mL). After 2 h, the reaction mixture was concentrated to a small volume on the rotavap and cooled in an ice bath before neutralizing with 1.0 N AcOH (0.70 mL). The solution was lyophilized to a crude product (0.1 g, 100%) as a white powder. ODS
chromatography (5% CH3CN/H20 cont~ining 0.1% TFA) afforded the title compound as a white powder MS (ES) m/e 358.4 (M+H)+.
Fxz~m~?le 37 Preparation of (S)-2.3.4.5-tetrahydro-7-~rr(4-aza-$-methylben7irr i~ ol-2-yl)methyllmethylaminolcarbonyll-3-oxo- lH- 1.4-benzodiazepine-2-acetic acid a) Dimethyl D-malate-o-trifluorom~th~npslllfonate A solution of dimethyl-D-malate (12.96 g, 80 mmol) and pyridine (6.8 mT., 84 mmol) in CH2Cl~ dry (50 mL) was added dropwise under argon at 0~C to a solution of trifluo~ nesulfonic anhydride (14.2 rnL, 84 mmol) in dry CH2Cl2 (40 rnL) in a flame dried flask. The resulting yellowish-orange mixture was stirred at 0~C for 30 rnin, and then at RT for 4 hr. The reaction was qllen~h~l by adding H20 (50 rnL), and the layers were separated. The organic layer was washed sequentially with H2O (3 x) and brine. Drying (MgS04) and concentration gave thetitle compound (22.45 g, 95%) as an off white solid: MS(ES) m/e 295 (M + H)+.
CA 0224l633 l998-06-26 W O 97/24119 PCTrUS96/20748 b) Dimethyl-N-(2-cyanophenyl)-D-aspartate A solution of dimethyl D-malate-O-trifluo~ n--sulfonate (22.4 g, 76.2 mmol) in CHCl3 (40 mL) and hexanes (40 mL) was added at to a solution of 2-aminobenzonitrile (9.0 g, 76.2 mmol) and 2,6-di-tert-butylpyridine in CHCl3 (50 mT) and hexanes (50 mL) in a flame dried flask at 0~C under argon. The resulting- mixture was stirred at 0~C for 30 min, then at RT for 3 days. The solvent was removed in vacuo and the residue was taken into EtOAc and washed sequentially with 5% HCI (10 x) and brine. Drying (MgSO4), concentration and silica gel flashchromatography (12% EtOAc/hexanes) gave the title compound (12.3 g, 62%) as a clear oil: MS(ES) mte 263.3 (M + H)+.
c) Methyl (S)-2,3,4,5-tetrahydro-3-oxo- lH- 1 ,4-benzodiazepine-2-acetate A mixture of dimethyl-N-(2-cyanophenyl)-D-aspartate ~12 g, 45.7 mmol), Et3N (7.64 mL, 54.84 mmol) and Raney-Ni (46 g, wet, prewashed with CH30H) in CH30H (200 mL) was stirred at RT under H2 (balloon) for 2 days. The catalyst wasremoved by filtration and washed with CH30H (3 x). Concentration and silica gel flash chromatography (0-5% CH3OH/CH2Cl2) gave the title compound (7.93 g, 74 %) as a white solid: MS(ES) m/e 235.3 (M + H)+.
d) Methyl (S)-2,3,4,5-tetrahydro-7-bromo-3-oxo- 1 H- 1 ,4-benzodiazepine-2-acetate Tetrabutylammonium tribromide (5.16 g, 10.7 mmol) was added portionwise to a solution of methyl (S)-2,3,4,5-tetrahydro-3-oxo-lH-1,4-benzodiazepine-2-acetate (2.5 g, 10.7 mmol) in CHCl3 (50 mL), and the llliY~tUl'e was stirred at RT for 2 days. H2O (30 mL) was then added, and the organic layer was separated and washedsequentially with H20 and brine. Drying (MgSO~), concentration, and silica gel flash chromatography (0-5% CH3OH/CH2Cl2) gave the title compound (1.99 g, 60%)) as a white solid: MS(ES) m/e 313.0 (M + H)+.
e) Methyl (S)-2,3,4,5-tetrahydro-7-[t[(4-aza-5-methylbeu7i.l.i~l~7Ol-2-yl)methyl]methylamino]carbonyl]-3-oxo- 1 H- 1 ,4-benzodiazepine-2-acetate A ~ tule cont~inin~ methyl (S)-2,3,4,5-tetrahydro-7-bromo-3-oxo-lH-1,4-benzodiazepine-2-acetate (624 mg, 2 mmol), 2-(aminomethyl)4-a_a-5-methylben7imicl~7ole dihydrochloride (695 mg, 2.8 mmol), DIEA (1.8 mL, 10 mmol), and (Ph3P)2PdCl2 (126 mg, 0.18 mrnol) in NMP (22 mL) was heated to 110~C under a CO balloon for 48 hr. The solvent was removed on the rotavap (highvacuum) and the residue was puri~led by silica gel flash chromatography (0.5 - 5%
W O 97/24119 PCTrUS96120748 CH3OEI/CH2Cl2) to give the title compound (170 mg, 19.5%) as a pale yellow solid: MS (ES) m/e 437.5 (M + H)'.
f~ (S)-2,3,4,5-Tetrahydro-7-[~[(4-aza-5-methylben7imitlz-~ol-2-5 yl)methyl]methylamino]carbonyl]-3-oxo- 1 H- 1,4-benzodiazepine-2-acetic acid 1.0 M LiOH (0.6 mL, 0.6 mmol) was added dropwise to a solution of methyl (S)-2,3,4,5-tetrahydro-7-[[[(4-aza-5-methylbçn~.imi~1~7.ol-2- -yl)methyl}methylamino]carbonyl]-3-oxo-lH- 1,4-benzodiazepine-2-acetate (170 mg, 0.39 rnmol) in CH30H (5 mL) and THF (5 mL)at RT. The resulting rnixture was 10 stirred for 20 hr and then was concenlldted. The residue was dissolved in H20, acidified with 30% TFA, and purified by ODS chromatography (5% CH3CN/H20 contz-ining 0.1% TFA). Concentration and lyophilization gave the title compound as an off white powder: [a],,25 -74.5o (c = 1, CH30H); MS (ES) m/e 423.2 (M + H)~.
Anal. Calcd for C2,H22N604 ~ 2 TFA ~ 1.75 H20: C, 44.03; H, 4.06; N, 12.32. Found:
C, 44.33; H, 4.04; N, 12.28.
Fx~nlple 38 Plep~dlion of (S)-2.3~4.5-tetrahydro-7-rrr(ben7imitl~zo1-2-yl)metbyl~minolcarbonyll-3-oxo- 1 H- 1 ~4-benzodiazepine-2-acetic acid a) Methyl (S)-2,3,4,5-tetrahydro-7-iodo-3-oxo- l H- 1,4-benzodiazepine-2-acetatePyridine-ICI complex: Iodine monochloride (100 mL, 1 M solution in CH2Cl2) was added slowly to a solution of pyridine (8.5 mL, 105 mmol) in dry CH2Cl2 (20 mL) at 5~C under argon so that the temperature was m~int~in~d between10 - 15~C. The mixture was stirred at 5 - 10~C for 20 min, then hexanes (50 mL) was added, and the rnixture was stirred in the cold bath for another 30 min. Thesolid was collected by suction filtration, washed sequentially with hexanes and petroleum ether, and dried to afford the reagent (22.5 g) as a yellow, crystalline solid.
Pyridine-ICl complex (1.27 g, 5.28 mmol) was added portionwise to a solution of methyl (S)-2,3,4,5-tetrahydro-3-oxo-lH-1.4-benzodiazepine-2-acetate (1.18 g, 4.8 mmol) in CH2Cl2 (20 rnL) and CH30H ~20 mT). The resulting mixture was stirred at RT for 40 rnin, then l M NaHSO3 (20 mL) was added. The solid was collected by suction filtration and washed with Et2O. Drying yielded the title compound (1.72 g, qu~ntit~tive) as an off-white solid: MS (ES) m/e 361.2 (M + H)~.
b) Methyl (S)-2,3,4,5-tetrahydro-7-r[[(ben7imi~ 7OI-2-yl)methyl]amino]carbonyl]-3-oxo- l H- 1,4-benzo~ 7~pine-2-acetate A mixture of methyl (S)-2,3,4,5-tetrahydro-7-iodo-3-oxo-lH-1,4-benzodia_epine-2-acetate (1.08 g, 3 mmol), 2-aminomethylben7imi~1~7Ole S dihydrochloride hydrate (924 mg, 4.2 mmol), DIEA (2.6 mL, 15 mmol), and (Ph3P)2PdCl2 (211 mg, 0.3 mmol) in NMP (30 mL) was heated to 110~C under a CO
- balloon for 3 hr. The solvent was removed on the rotavap (high vacuum) and the residue was purified by silica gel flash chromatography (0-7% CH30H/CH2Cl2) to afford the title compound (530 mg, ~44%) as an off white solid: MS(ES) m/e 408.1 (M+H)t.
c) (S)-2,3,4,5-Tetrahydro-7-{[[(ben7imi~ 7ol-2-yl)methyl]amino]carbonyl]-3-oxo-1 H- 1,4-benzodiazepine-2-acetic acid Following the procedure of Example 37(f), except substituting methyl (S)-2,3,4,5-tetrahydro-7-[[[(ben7imi(1z~7Ol-2-yl)methyl]amino]carbonyl]-3-oxo-lH-1,4-benzodiazepine-2-acetate for methyl (S)-2,3,4,5-tetrahydro-7-[[[(4-aza-5-methylbenzimidazol-2-yl)methyl]methylamino3carbonyl]-3-oxo- lH- 1,4-benzodiazepine-2-acetate, the title compound (66%) was ~ al~,d as a white powder: [a]n25 -145.3~ (c = 1, CH30H); MS (ES) m/e 394.2 (M + H)t. Anal. Calcd for C2nH,gN5O4 - 2 TFA 0.125 H2O: C, 46.22; H, 3.43; N, 11.23. Found: C, 46.13;
H, 3.78; N, 11.49.
E~am~l?le 39 ~lGpaldlion of (+)-N-r2-(aminomethyl~-4-rrr~4-~7~-S-methylbe~7,imi-1~7ol-2-yl)methyl~methylaminolcarbonyllphenyl~aspartic acid a) Methyl (+)-2,3,4,5-tetrahydro-7-[[[(4-a_a-5-methylben7imi~1~7ol-2-yl)methyl]methylamino]carbonyl3 -3-oxo- 1 H- 1,4-benzodia_epine-2-acetate EDC (130 mg, 0.75 mmol) was added to a solution of methyl (+)-2,3,4,5-tetrahydro-7-carboxy-3-oxo-lH-1,4-benzodiazepine-2-acetate (190 mg, 0.68 mmol), 2-(aminomethyl)-4-aza-S-methylben7imi~l~7ole dihydrochloride (169 mg, 0.68 mmol), HOBt H2O (101 mg, 0. 75 mmol), and DIEA (0.39 mL. 2.24 mmol) in anhydrous DMF at RT. After 20 hr, the reaction was concentrated on the rotavap (high vacuum), and the residue was cl~ atographed on silica gel (1 - 6.5%
CH3OH/CH2Cl2) to afford the title compound (260 mg, 88%) as a white solid: MS
(ES) m/e 437.5 (M ~ H)'.
b) (_)-N-[2-(Aminomethyl)-4-[[[(4-aza-5-methylben7imitl~7O1-2-yl)methyl]methylamino}carbonyl]phenyl]aspartic acid Following the procedure of Example 37(f), except substitl-ting methyl (+)-2,3,4,5-tetrahydro-7-[[[(4-~a-5-methylben7imi~1~7O1-2-yl)methyl]methylamino]carbonyl]-3-oxo-lH-1,4-ben7O~ 7~pine-2-acetate for methyl (S)-2,3,4,5-tetrahydro-7-[[[(4-aza-5-methylben7imi~1~701-2-yl)methyl]methylamino]carbonyl]-3-oxo- 1 H- l ,4-bçn70~ 7epine-2-acetate, the title compound (66%) was prepared as a white powder: MS (ES) m/e 441.2 (M + H)+.
Anal. Calcd for C2,H2"N6O5 - 2 TFA 2.25 H2O: C, 42.08; H, 4.38; N, 11.78. Found:C, 42.01; H, 4.18; N, 11.55.
Fx~rru?le 40 Plepaldlion of (S~-2~3.4.5-tetrahydro-7-lrr(4-aza-5-methylbell7i.. i~zl7ol-2-yl)mt~thyll~minolcarbQ~.~yll-3-oxo-lH-1~4-benzodiazepine-2-acetic acid a) Methyl (S)-2,3,4,5-tetrahydro-7-[[[(4-aza-5-methylberl 7i mi~l ,.7 ol-2-yl)methyl]amino]carbonyl]-3-oxo- I H- 1,4-benzodiazepine-2-acetate Following the procedure of Example 38(b), except ~7u1)~7LiLu~il.g 2-(aminomethyl)-4-aza-5-methylbell,i..,i-lz~7Ole dihydrochl(-ride for 2-aminomethylbe~7imicl~7ole dihydrochloride hydrate, the title compound (63%) was prepared as an amber solid: MS ~ES) m/e 423 (M + H)'.
b) (S)-2,3,4,5-Tetrahydro-7-[[[(4-aza-5-methylben7imicl~7O1-2-yl)methyl]amino]carbonyl~-3-oxo- 1 H- 1,4-benzodiazepine-2-acetic acid Following the procedure of Example 37(f), except substituting methyl (S)-2,3,4,5-tetrahydro-7-[[[(4-aza-5-methylb~n7imi~ 7ol-2-yl)methyl]amino]carbonyl]-3-oxo- lH- 1,4-benzodiazepine-2-acetate for methyl (S)-2,3,4,5-tetrahydro-7-t[[(4-aza-5-methylbe~-7i.. i-l~7~-1-2-yl)methyl]methylamino]~ul,o.,yl]-3-oxo-lH-l,~benzodiazepine-2-acetate, the title colll~oulld (50%) was prepared as a white powder: MS (~S) m/e 409.2 (M + H)~. Anal. Calcd for C2"H2"N6O4 ~ 1.75 TFA -H~O: C, 45.09; H, 3.82; N,3 .45. Found: C, 45.18; H, 4.10; N, 13.58.
E~mple 41 W O 97/24119 PCTrUS96/20748 Preparation of (S)-2.3.4.5-Tetrahydro-7-rrr(bPnzimitl:~7ol-2-yl)methyllaminolcarbonyll-3-oxo-4-r2-(pyrid-3-yl)ethyll-lH- 1.4-ben7-1diazepine-2-acetate 5 a) tert-Butyl 4-fluoro-3-[[2-(pyrid-3-yl)ethyl~amino]benzoate A mixture of tert-butyl 4-fluoro-3-methylbenzoate (3.83 g, 18.22 mmole), - NBS (3.57 g, 20.24 mmole), benzoyl peroxide (0.22 g, 0.91 mmole), and CC14 (90 mL) was heated at reilux. After 16 hr, the reaction was cooled in ice/H20 and filtered, and the filtrate was concentrated. The residue was passed through a short 10 pad of silica gel (20 % EtOAc/hexane) to remove baseline m~t~.ri~ls~ and the filtrate was concentrated. The residue was dissolved in THF ~90 mI,), and 3-(2-aminoethyl)pyridine (6.97 g, 57 mmol).) was added rapidly. The addition a~eal ,dto be mildly endothermic. The reaction was stirred overnight then was concentrated.
The residue was diluted with Et2O (100 mL) and washed sequentially with 1.0 N
15 NaOH (30 mL), H2O (30 mT.), and brine (30 mL). Drying (MgSO4), concentration,and silica gel chromatography (10 % MeOH/CH2Cl2) gave the title compound (2.58 g, 59 %) as a yellow oil: MS (ES) m/e 331 (M + H)+.
b) tert-Butyl (S)-4-fluoro-3-[2-aza-4-(benzyloxycarbonyl)amino-3,6-dioxo-6-20 methoxy-2-[2-(pyrid-3-yl)ethyl]hexyl]benzoate DCC (1.86 g, 9 mmol)was added to a solution of tert-butyl 4-fluoro-3-[[2-(pyrid-3-yl)ethyl]amino]benzoate (2.7 g,.8.18 mmol), N-Cbz-L-aspartic acid ~-methyl ester (J. Am. Chem. Soc. l9S7, 79, 5697; 2.53 g, 9 mmol.), and HOBt H2O
(1.2 g, 9 mmol) in anhydrous DMF (10 mL) at RT. After 24 hr, the mixture was 25 diluted with Et20 (25 rnL) and filtered. The filtrate was concentrated to dryness, and the residue was diluted with Et2O (50 rnL) and washed with H2O (2 x 10 mL) and brine (10 mL). Drying (MgSO4), concentration, and silica gel chromatography (CH2Cl2) gave the title compound (2.4 g, 49%)as a colorless oil: MS (ES) m/e 594(M +H)+.
c) tert-Butyl (S)-4-fluoro-3-[4-amino-2-aza-3,6-dioxo-6-methoxy-2-[2-(pyrid-3-yl)ethyl]hexyl]benzoate A mixture of tert-butyl (S)-4-fluoro-3-[2-aza-4-(benzyloxycarbonyl)arnino-3,6-dioxo-6-methoxy-2-[2-(pyrid-3-yl)ethyl]hexyl]benzoate (2.4 g, 4 mmol.), 10 %35 Pd/C (184 mg, 0.17 mmole), and MeOH (17 mL) was shaken at RT under H2 (50 psi). After 1.5 hr, the reaction was filtered through celite(~3) and concentrated. Silica W O 97/24119 PCT~US96/20748 gel chromatography (10 % MeOH in 1:1 EtOAc/CHC13) gave the title compound (1.1 g, 59 %) as a colorless oil: MS (ES) 460 (M + H)+.
d) Methyl (S)-2,3,4,5-tetrahydro-7-(tert-butoxycarbonyl)-4-[2-(pyrid-3-yl)ethyl]-3-5 oxo- lH- 1,4-benzodiazepine-2-acetate A solution of tert-butyl (S)-4-fluoro-3-t4-amino-2-aza-3,6-dioxo-6-methoxy-2-[2-(pyrid-3-yl)ethyl]hexyl]ben70~te (0.64 g 1.39 mrnol) in anhydrous DMSO (5.7mL) was heated under argon in an oil bath set at 120-125~C. After 17.5 hr, the reaction was cooled in ice/H20 and~diluted with H20 (12 mL). The mixture was 10 extracted with EtOAc (3 x 20 mL), and the combined EtOAc layers were washed with H2O (10 mL) and brine (10 mL). Drying (MgSO4), concentration, and silica gel chromatography (9: 1 CH2Cl2/MeOH) gave the title compound (0.15 g, 33%) as anearly colorless solid: MS (ES) m/e 440 (M +H)+.
e) Methyl (S)-2,3,4,5-tetrahydro-7-carboxy-4-[2-(pyrid-3-yl)ethyl]-3-oxo-lH-1,4-benzodiazepine-2-acetate 4 M HCI/dioxane (0.5 mL) was added to a solution of methyl (~)-2,3,4,5-tetrahydro-7-(tert-butoxycarbonyl)-4- [2-(pyrid-3-yl)ethyl]-3 -oxo- 1 H- 1,4- -benzodiazepine-2-acetate(0.18 g, 4.1 mrnol) in anhydrous CH2Cl2 (5 mL) and the reaction was stirred at RT overnight. Concentration in vacuo followed by reconcentration from toluene (3 x 10 mL) gave the title compound (0.12 g, 65%):
MS (ES) m/e 384 (M + H)+.
f) Methyl (S)-2,3,4,5-tetrahydro-7-[t[(ber 7imitl~7ol-2-yl)methyl]amino]carbonyl]-3-oxo-4-[2-(pyrid-3-yl)ethyl]- 1 H- 1,4-benzodiazepine-2-acetate A mixture of methyl (S)-2,3,4,5-tetrahydro-7-carboxy-4-[2-(pyrid-3-yl)ethyl]-3-oxo-lH-1,4-benzo~ 7epine-2-acetate (0.12 g, 0.26 mmol) and thionyl chloride (15 mL) was refluxed for 1 h. The re~slllting orange solution was concentrated to dryness to leave a yellow-orange foam. This was dissolved in CH2C12 (10 mL) and added dropwise to a solution cont~inin~ 2-aminomethylbe..~ilni~l~701e dihydrochloride hydrate (0.058 g, 0.26 mmol), pyridine (0.72 g, 9.1 mmol), and triethylamine (0.55 g, 5.46 mmol) in CH2C12 (15 mL) at 0~C under argon. The reaction mixture was then stirred in RT under argon. After 25.5 h, CH2C12 (200 mL) and 5% NaHCO3 (50 rnL) were added to the reaction 35 mixture to give a light yellow precipitate which was filtered and air-dried to give the title compound (0.030g.,~% yield): MS (ES) m/e 513 (M+H)+.
W O 97/24119 PCTrUS96/20748 g) (s)-2~3~4~5-Tekahydro-7-~[[(ben~ 7ol-2-yl)methyl]amino]carbonyl] -3-oxo-4-[2-(pyrid-3-yl)ethyl]- 1 H- 1,4-benzodiazepine-2-acetate 1.0 N LiOH (0.57 mL, 0.57 mmol) was added dropwise at RT to a mixture of S methyl (S)-2,3,4,5-tetrahydro-7-[[[(ben7imi(1~7~-1-2-yl)methyl3amino]carbonyl]-3-oxo-4-[2-(pyrid-3-yl)ethyl]-lH-1,4-benzodia_epine-2-acetate (0.030 g, 0.059 mmol) - in THF (4 mL) and H2O (5 mL). The resulting light brownish-yellow solution was stirred for 21.5 h, then was concenkated on the rotavap. The resulting residue was lyophilized to give the crude product as a yellowish powder. ~ liv~ HPLC
(PRP-l~ column, step gradient, 10-20% CH3CN/H2O contzlinin~ 0.1% TFA) afforded the title compound (0.OlOg 34% yield): MS (ES) m/e 499(M+H)+. Anal.
Calcd .for C27H26N6O4 - 3 C2HF3O2 ~ 3 HCI - 3 H2O: C, 37.41; H, 3.41; N,7.52.
Found: C, 37.6; H, 3.52; N, 7.52.
Example 42 Preparation of Ethyl (S)-2.3.4.5-tetrahydro-7-rrr~ben7imid~7ol-2-yl)methyl~methylaminolcarbonyll-4-methyl-3-oxo- 1 H- 1.4-benzodiazepine-2-acetate a) Ethyl (s)-2~3~4~s-tekahydro-7-~[[(be~ 7~l-2-yl)methyl]methylamino]carbonyl]-4-methyl-3-oxo- lH- 1,4-benzodia_epine-2-acetateHCl gas was bubbled into EtOH (200 mL) for 10 min, then (S)-2,3,4,5-tetrahydro-7-[[[(ben7imi~ 7ol-2-yl)methyl]methylamino]carbonyl]~-methyl-3-oxo-lH-1,4-benzodia7epine-2-acetic acid (2.00 g, 4.5 mmol) was added. The reaction was stirred at RT for 24 hr, then was concentrated to dryness on the rotavap. The residue was reconcenkated from toluene (2 x) to remove residual EtOH, then was chromatographed on silica gel (gradient: 7% MeOH/CH2Cl2 (1 L) then 10%
MeOH/CH2Cl2). The r~sllltin~ residue was dissolved in EtOH, and Et2O was added to precipitate a solid. This was collected and washed with Et2O to afford the title compound as a white solid: MS (ES) m/e 450.2 (M + H)+. Anal. Calcd for C24H27NsO4 ~ 1.5 H2O: C, 60.49; H, 6.35; N, 14.70. Found: C, 60.41; H, 6.27; N, 14.38.
Fx~mple 43 Preparation of 4-~rr3-(lH-ben7imi~1~7ol-2-yl)propyllaminolcarbonyllpiperidine-l-acetic acid W O 97/24119 PCTrUS96/20748 a~ 2-[3-~N-tert-Butoxycarbonyl)aminopropyl3benzirnidazole A solution of isobutylchloroformate ( 10.2 rnL, 79 mmol) in THF (25 rnL) was added to a solution of 4-(tert-butoxycarbonyl)arninobutyric acid (Organic S ~;ynthesis 1984, 63, 160; 13.5 g, 0.066 mole) and triethylamine (11 mL, 80 mmol) in THF ( 50 mL) at 0~C under argon. After 0.5 h, a solution of 1,2-phenylen~ mine (7 g, 64.8 rnmole) in THF (50 rnL) was added dropwise to the res-ltting white suspension. The reaction was stirred for 18 h, then was ffltered, and the filtrate was concentrated to give a serni-solid. This was dissolved in AcOH (100 rnL), and the solution was heated at 70~C for 18 h. The reaction mixture was concentrated, andthe residue was reconcentrated several times from toluene. Silica gel flash chromatography gave the title compound (6.0 g, 33%): MS (ES) m/e 276 [M+H]t.
b) 2-(3-Aminopropyl)bc~.~i...itl~7ole dihydrochloride A solution of 2-[3-(N-tert-butoxycarbonyl)aminopropyl3ben7imirls-7.nle (1.2 g, 4.3 mmol) and 4 M HCl in dioxane (20 mL) in CH2C12 (25 ml) was stirred at RT
for 18 h. The resulting white suspension was filtered to give the title compound(1.07 g, 97%).
20 c) Ethyl ~[[[3-(lH-ben,.iid~7.ol-2-yl)propyl]arnino]carbonyl]piperidine-l-acetate A ~ lule of ethyl 4-carb-~y~ 3elidine-1-acetate hydrochloride (Yellin's SB
223913 CIP)(0.76 g, 3 mmol) in thionyl chloride (10 mL) was heated to reflux for15 min, then was concentrated to dryness. After evaporation several times from toluene, the residue was dissolved with 2-(3-aminopropyl)ben7imi~1~7Ole dihydrochloride (0.77 g, 3 mmol) and DIEA (3 mL) in DMF (25 mL). After 18 h, the reaction mixture was partitioned between EtOAc (50 mL) and 5% NaHCO3 (100 mL), and extracted with EtOAc (2 x 50 mL). The combined organic extracts were washed seq~lenti:~ily with H20 and saturated NaCl solution, then were dried overMgSO4. Concentration and silica gel flash chromatography (6% MeOH/CH2C12) gave the title compound (40 mg, 3%).
b) 4-[[~3-(Rçn7.imida_ol-2-yl)propyl]amino~carbonyl]piperidine-1-acetic acid lN NaOH solution (0.4 mL, 0.4 mmol) was added to a stirred solution of ethyl 4-[[3-~lH-ben7imi~l~7ol-2-yl)propylamino]carbonyl]piperidine-l-acetate (40mg, 0.1 mmol) in MeOH (10 mL)at RT. After 18 h, the mixture was neutralized with AcOH (1 mL) and concentrated to remove the MeOH. The aqueous solution was loaded onto an XAD-2 column, and eluted with H20 (500 mL), then with 20%
W O 97/24119 PCTrUS96/2074P
CH3CNt H20. The fractions con,~zlining the product were pooled and lyophilized to give the title compound (9 mg, 25%). MS (ES~ m/e 345.2 [M+H]+.
Anal. Calcd for C,8H24N4O3 0.75 H2O: C, 60.40; H, 7.18; N, 15.65. Found: C, 60.48; H, 7.16; N, 15.40.
Lxample 44 Plc~aldliQn of 4-rrr3-(ben7imi~l~7f 1-2-yl)propyllaminolcarbonyllphenylacetic acid a) Ethyl 4-[[[3-~ben7imiclz~ 1-2-yl)propyl]amino]carbonyl]-l-phenylacetate A mixture of ethyl 2-(4-carboxyphenyl)acetate (Yellin's SB 223913 CIP)(0.5 g, 2.4 mmol), 2-(3-aminopropyl)ben7imi~7ole dihydrochloride (0.7 g, 2.8 mmol), HOBt ~ H20 ~0.36 g, 2.6 mmol), EDC (0.5 g, 2.6 mmol), and DIEA (1.5 mL, 8.8 mmol) in DMF (15 mL) was warmed briefly and stirred at RT for 18 h. The reactionmixture was partitioned between EtOAc ~50 mL) and 5% NaHCO3 (100 mL), and extracted with EtOAc (2 x 50 mL). The combined organic extracts were washed sequentially with H2O and saturated NaCI solution, then were dried over MgSO4.
The evaporated residual solid was triturated with Et2O to give the title compound (0.56 g, 66%): MS (ES) m/e 366.0[M+H~+. Anal. Calcd for C2lH23N3O3 0.25 H2O:
C, 68.18; H,6.40; N, 11.36. Found: C, 68.16; H,6.26; N,11.36.
b) 4-[[[3-(Ben7imi~1~7(-1-2-yl)propyl]amino]carbonyl]phenylacetic acid 1 N NaOH solution (6 rnL, 6 mmol) was added to a stirred solution of ethyl 4-[[[3-(lH-bPn7imicl~7n-2-~71)propyl]arnino]carbonyl]-l-phenylacetate (0.38 g, 1 25 mmol) in MeOH (15 mL)at RT. After 4 h, the mixture was neutralized with AcOH
(6 mL) and the resulting solid was filtered to give the title compound (77 mg, 22%):
MP 108-110~C; MS (ES) rn/e 345.2 [M+H]+. Anal. Calcd for Cl9HlgN3O3 0.6 H2O:
C, 65.54; H, 5.85; N, 12.07. Found: C, 65.63; H, 5.65; N, 11.95.
Fx~m~le 45 Preparation of (S)-2.3.4.5-tetrahydro-4-m~t~yl-3-oxo-7-rrr(S-trifluoromethylben~ 7ol-2-y~)methyl~methylaminolcarbonyll - l H- l .4-benzodiazepine-2-acetic acid a) 3,4-Diaminobenzotrifluoride W O 97~4119 PCT~US96/20748 4-Amino-3-nitroben~oLlinuoride (3.7070 g, 17.98 mmol) was dissolved in MeOH, and a catalytic amount of 10% Pd/C was added. The reaction was purged with H2, and stirred at RT under H2 (balloon). After 24 hr, the reaction was filtered through a bed of celite~, and the filtrate was evaporated under vacuum to yield the title compound (3.0878 g, 98%). The material was used without characlefizdLion.
b) 2-[N-(Benzyloxycarbonyl)-N-methyl]aminomethyl-5-trifluoromethylben7imi~ 1e Cbz-sarcosine (3.9950 g, 17.13 mmol) was dissolved in dry THF, and isobutylchloroformate (2.5 mL, 19.27 mmol) was added, followed by triethylamine (5.0 mL, 35.95 mmol). The mixed anhydride was allowed to form at RT for 30 min, then was added to a solution of 3,4-diaminobenzotrifluoride (3.0818 g, 7.53 mmol) in dry THF. After 20 hours at RT, the reaction was ~vapoldted under vacuum. The residue was partitioned between EtOAc and 1.0 N NaHCO3, and the layers were separated. The aqueous layer was extracted with EtOAc, and the combined organic layers were dried (MgSO4), filtered and e-v~oldled under vacuum. The residue wasreconcentrated from toluene, then was dissolved in glacial AcOH (125 mL). The solution was heated at 110~C for 24 hr, then the AcOH was evaporated under vacuum. The residue was reconcentrated from toluene, then was adsorbed onto silica gel and loaded onto a dry silica gel flash chromatography column. The column was eluted with 1:1 CHCl3/Et,0 to afford the title compound (2.9397 g, 47.2%): TLC R, (1:1 CH2Cl~/Et2O) 0.57; MS (ES) rn/e 364.2 (M+H)+; 'H NMR (250 MHz, CDC13) ~ 8.0 - 7.2 (m, 9H), 5.05 (s, 2H), 4.84 (s, 2H), 3.07 (s, 3H).
c) 2-(Methylaminomethyl)-5-trifluoromethylben7.imicl~701e 2-[N-(Benzyloxycarbonyl)-N-methyl]aminomethyl-S-tri~uoromethylben7imi(1~7ole (2.9397 g, 8.09 mmol) was dissolved in MeOH, and a catalytic amount of 10% Pd/C was added. The reaction was purged with H2, then was stirred at RT under H2. After 5 hr, the reaction ~ lure was filtered through a bed of celite(~), and the filtrate was evaporated under vacuum to leave a tan colored oil. Analysis by 400 MHz NMR showed the Cbz protecting group was still present, so the residue was resubmitted to the reaction conditions. After 18 hr, the catalyst was removed by filtration through a bed of celite(~) and the filtrate was ~;v~oldted under vacuum to yield the title compound (1.7809 g, 96%): lH NMR (250 MHz, CDC13) o 7.76 - 7.32 (m, 4H), 4.32 (s, 2H), 2.59 (s, 3H).
W O 97/24119 PCT~US96/20748 d) Methyl (S)-2,3,4,5-tetrahydro4-methyl-3-oxo-7-[[[(5-trifluoromethylben7im~ 7Ol-2-yl)methyl]methylamino]carbonyl]- lH- 1,4-benzodiazepine-2-acetate Methyl (S)-7-carboxy-4-methyl-3-oxo-2,3 ,4,5-tetrahydro- l~I- 1,4-S benzodiazepine-2-acetate (179.2 mg, 0.61 mmol) was weighed into a 100 mL round bottomed flask. CH3CN (10 mL) was added, followed by HOBt ~ H2O (97.9 mg, 0.72 mmol) and EDC (149.3 mg, 0.78 mmol). After all the solids had dissolved, a solution of 2-(methylarninomethyl)-5-trifluoromethylben7imi~1~701e (186.1 mg, 0.81 mmol) in CH3CN was added with diisopropylethylamine (0.25 rnL, 1.44 mmol).
After 24 hr at RT, the reaction was evaporated under vacuum, and the residue waschromatographed on silica gel (3% MeOH/CHCl3) to afford the title compound (308.1 mg, 100%): TLC Rf (5% MeOH/CHCl3) 0.21; lH NMR (250 MHz, CDC13) 7.83-7.16 (m, 7H), 5.37 (d, 1~), 5.05-4.70 (m, 3H), 2.96 (m, 3H), 3.72 (s, 3H), 3.16 (s, 2H), 2.11 (s, 3H); MS (ES) m/e 504.0 (M+H)~.
e) (s)-2~3~4~5-Tetrahydro-4-methyl-3-oxo-7-[[[(s-trifluoromethylben~ 7Q1-2 yl)methyl]methylamino]carbonyl] -1 H- 1 ,4-benzodiazepine-2-acetic acid Methyl (S)-2,3,4,5-tetrahydro-4-methyl-3-oxo-7-[[[(5-trifluoromethylb~n7imi~1~7ol-2-yl)methyl]methylamino~carbonyl]-lH-1,4-benzodiazepine-2-acetate (308.1 mg, 0.61 mmol) was dissolved in MeOH (S mL).
H20 (5 mL) was added, followed by 1.0 N NaOH (2.0 mL, 2.0 mmol). After 24 hr at RT, the reaction was neutralized with 1.0 N HCl (2.0 mL). The milky white mixture was stirred at RT for 15 min, then was diluted with H2O, and the precipitate was collecte~l on a sintered glass funnel. The white powder was dried in a vacuum de~ c~tQr overnight to yield the title compound (268.0 mg, 90%): M~ (ES) m/e 490.2 (M+H)'. Anal. Calcd for C23H~N5O4F3 2.25 H2O ~ 0.25 HCl: C, 51.24; H, 5.00; N, 12.99. Found: C, 51.44; H, 4.96; N, 12.45.
F.x:~ml?le 46 Ple~ alio l of (S)-2.3.4.5-tetrahydro-7-rrr(4.7-dimethoxyben7imicl~7.ol-2-yl)methyllmethylamino~carbonyll-4-methyl-3-oxo- lH- 1 .4-benzodiazepine-2-aceticacid ~' a) 2-[N-(Benzyloxycarbonyl)-N-methyl]~n~innmPthyl-4,7--1im~thoxybenzimidazole ~ Following the procedure of Example 45(b), except sul)~lilulillg 1,2-~ mino-3,6-dimethoxybenzene for the 3,4-diaminobenzotrifluoride, the title compound was 167 W O 97~4119 PCT~US96~0748 prepared: MS (ES) m/e 356.2 (M+H)'; lH NMR (250 MHz, CDC13) o 7.34 (s, SH), 6.54 (d, 2H), 5.18 (s, 2H), 4.65 (s, 2H), 3.95 (s, 3H), 3.86 (s, 3H), 3.03 (s,3H).
S b) 4,7-Dimethoxy-2-(methylaminomethyl)ben7imi(1~7ole 2-[N-(Benzyloxycarbonyl)-N-methyl]aminomethyl-4,7-dimethoxybenzimidazole (186.5 mg, 0.53 mmol) was dissolved in MeOH, and a catalytic amount of 10% Pd/C was added. The reaction was purged with Hz, then was stirred at RT under H~ (balloon). ~After 20 hr, the reaction was filtered through celite(~, and the filtrate was eva~?oraLed under vacuum to yield the title compound (96.9 mg, 83%): 'H NMR (250 MHz, CDC13) ~ 6.52 ~s, 2H), 3.94-3.86 (m, 6H), 2.36 (s, 3H).
c) Methyl (S)-2,3,4,5-tetrahydro-7-[[[(4,7-~lim~thQxyben7imifl~7ol-2-yl)methyl]methylamino]carbonyl]4-methyl-3-oxo- 1 H- 1,4-benzodiazepine-2-acetate Methyl (S)-7-carboxy~-methyl-3-oxo-2,3,4,5-tetrahydro-lH-1,4-benzodiazepine-2-acetate (112.3 mg, 0.38 rnmol) was weighed into a 100 mL round bottomed flask. CH3CN was added, followed by HOBt ~ H~O (62.3 mg, 0.46 mmol~
and EDC (120.0 mg, 0.63 mmol). When the solids had all dissolved, diisopropylethylamine (0.1 mL, 0.57 mmol) was added, followed by a suspension of4,7-~lim~thoxy-2-(methylaminomethyl)be~-7i~ 7ole (96.8 mg, 0.44 mmol) in CH3CN cont5.inin~ diisopropylethylamine (0.1 mL, 0.57 mmol). After 2.5 days at RT, the reaction was evaporated under vacuum. The residue was evaporated once with toluene, then was chromatographed on silica gel (CHCl3, then 5%
MeOH/CHCl3) to afford the title compound (152.0 mg, 80.0%): TLC R~ (5%
MeOH/CHCl3) 0.35; MS (ES) m/e 496.2 (M+H)t; H NMR (250 MHz, CDC13) o 7.25 (m, 2H), 6.56 (s, 2H), 5.36 (d, lH), 3.91 (s, 6H), 3.70 (s,3H), 3.08 (s, 3H).
d) (S)-2,3,4,5-Tetrahydro-7-[[[(4,7-dimetho~ybel-7il..i~1~7~ 2-yl)methyl]methylarnino]carbonyl]-4-methyl-3-oxo- 1 H- 1,4-benzodiazepine-2-acetic acid Following the procedure of Example 45(e), methyl (S)-2,3,4,5-tetrahydro-7-t[[~4,7-dimethoxybçn7.imic~7.(-1-2-yl)methyl]methylamino]carbonyl]-4-methyl-3-oxo-lH-1,4-benzodiazepine-2-acetate was saponified to afford the title compound (110.0 mg, 74%): MS (ES) m/e 482.2 (M+H)~. Anal Calcd for C~,~H27N5O6 ~ 0.75 H2O: C, 58.23; H, 5.80; N, 14.15. Found: C, 58.26; H, 5.59; N, 13.90.
Fxample 47 ~l~dlion of (S)-2.3.4.5-tetrahydro-7-rrr(4-methylbenzimidazol-2-yllmethyllmethylaminolcarbonyll-4-methyl-3-oxo-lH-1.4-benzodiazepine-2-acetic 5 a~id a) 1,2-Diamino-3-methylbenzene Following the procedure of Example 45(a), except ~ub~ li..g 2-methyl-6-nitro~nilin.- (3.0204 g, l9.g8 mmol) for the 4-amino-3-nitrobenzotrifluoride, the title compound (2.4815 g) was prepared. This was used without char~( le, i7~1 ;on.
b) 2-[N-(Benzyloxycarbonyl)-N-methyl]aminomethyl-4-methylben7imitl:~7ole C bz-sarcosine (4.6466 g, 19.92 mmol) was dissolved in dry THF in a 100 mL
round-bottomed flask. Triethylamine (3.0 mL, 21.57 mmol) was added, followed by isobutylchlolufol-llate (2.8 mL, 21.59 mmol). The white reaction mixture was stirred at RT for Q.5 hr, then was added to a solution of 1,2-diamino-3-methylbenzene (2.4815 g) in dry THF at -20 to -30~C. After 20 min, the reaction was warmed to RT and stirred there for 16 hr. The reaction was evaporated under vacuum and the residue was partitioned between EtOAc and 1.0 N NaHCO3. The layers were separated, and the aqueous layer was extracted with EtOAc. The combined organics were dried (MgSO4), filtered, and evaporated under vacuum. The residue was reconcentrated from toluene and the dried solid was dissolved in glacial AcOH (150 mL). The solution was heated at 110~C for 18 hr, then was concentrated under high vacuum. The residue was reconcentrated from toluene, then was adsorbed onto silica gel and loaded onto a dry silica ge} flash chromatography column. The column was eluted with 1: 1 CH2Cl2/Et2O to afford the title compound (3.1586 g, 51%): MS (ES) m/e 310.2 (M+H)~; 'H NMR (250MHz, CDCl3) ~ 7.35-7.01 (m, 10H), 5.00 ~s, 2H), 4.72 (s, 2H), 2.99 (s, 3H), 2.55 (s, 3H).
c) 4-Methyl-2-(methylaminomethyl)ben7imi<1~7Ole 30 Following the procedure of Example 45(c), except subslil~li.. g 2-[N-(ben_yloxycarbonyl)-N-methyl]aminomethyl-4-methylben7imit1~37.01e for the 2-[N-(benzyloxycarbonyl)-N-methyl]aminomethyl-5-trifluoromethylb~on7imi(1~7nle, the title compound (2.9916 g, qn~ v~) was prepared: 'H NMR (250 MHz, CDCl3) o 7.36-7.01 (m, 4H), 4.01 (s, 2H), 2.52 (s, 3H), 2.41 (s, 3H).
d) Methyl (S)-2,3,4,5-tetrahydro-7-[[[(4-methylben7imi~1~7ol-2-, yl)methyl]methylamino]carbonyl]-4-methyl-3-oxo- 1 H- 1,4-benzodi~epine-2-acetate W O 97/24119 PCTrUS96120748 Methyl (S)-7-carboxy-4-methyl-3-oxo-2,3,4,5-tetrahydro-lH-1,4-benzodiazepine-2-acetate (188.5 mg, 0.64 mmol) was weighed into a 100 mL round bottomed flask. CH3CN was added, followed, sequentially, by HOBt ~ H2O (103.5 mg, 0.77 mmol), EDC (149.3 mg, 0.78 mmol), and diisopropylethylamine (0.15 ml, 0.86 S mmol). After 15 min, a solution of 4-methyl-2-(methylaminomethyl)ben7imi~1~7ole (273.8 mg, 1.56 mmol) in CH3CN was added. CH2Cl2 (5 mL) was added to dissolve some material. After 18 hr at RT, the reaction was concentrated, and the residue was chromatographed on silica gel (5% MeOH/CHCl3) to afford the title compound (307.3 mg, q~l~ntifz~tive): MS (ES) rn/e 450.2 (M+H)t; 'H NMR (250 MHz, CDCl3) ~ 7.23-7.03 (m, 7H), 6.41 (br s, lH), 5.33 (d, J = 16.3 Hz, lH), 3.69 (s, 3H), 3.46 (s, 3H), 3.10 (s, 3H).
e) (S)-2,3,4,5-Tetrahydro-7-[[L(4-methylbçn7imidz-~ol-2-yl)methyl]methylamino]carbonyl3-4-methyl-3-oxo- lH- 1,4-benzodiazepine-2-acetic acid Following the procedure of Example 45(e), methyl (S)-2,3,4,5-tetrahydro-7-[[[(4-methylbçn7imicls-7ol-2-yl)methyl]methylamino]carbonyl]-4-methyl-3-oxo-lH-1,4-benzodiazepine-2-acetate ~307.3 mg, 0.68 mmol) was saponified to afford the title compound (243.9 mg, 82%): MS (ES) rn/e 436.2 (M+H)+. Anal Calcd for C23H25N504 -2.75 H2O: C, 56.96; H, 6.34; N, 14.44: Found: C, 56.72; H, 6.27; N, 14.26.
F.x~m~G~le 48 ~dLion of (S)-2.3.4.5-tetrahydro-7-rrr(4-aza-5.7-dim~tllylbe~ idazol-2-yl)methyllme~llylaminolcarbonyll-4-methyl-3-oxo- 1 H- 1.4-benzodiazepine-2-acetic a) 2-Amino-4,6-dimethyl-3-nitropyridine 2-Arnino-4,6-dimelhyl~ylidine (5.55 g, 45.43 mmol) was weighed into a 500 mL round bottomed flask. The flask was cooled to -78~C. Concentrated HzS04 (25 mL, 450 mmol) was added, followed by concentrated HNO3 (3.5 mL, 56.0 mmol).
The ~ui~Lul~ became a solid frozen mass. The cooling bath was removed and the reaction was allowed to warm to RT. After about 15 min there was an exothermic reaction with the release of some nitrous oxide gas, and the reaction became a very dark red color. The reaction was heated at 85 - 90~C for 3 hr, then was cooled to RT, diluted with ice, and neutralized with 6 N NaOH (160 mL). The aqueous solution was extracted with EtOAc (3 x), and the combined EtOAc layers were dried .
W O 97/24119 PCTrJS96/20748 (MgSO4), filtered, and evaporated under vacuum. The resulting yellowish-orange solid was adsorbed onto silica gel and flash chromatographed on a dry silica gel colurnn. The column was eluted with 1: 1 CHCI3/Et2O to afford the title compound (1.0650 g, 14%): MS (ES) m/e 168.0 (M+H)'.
s b) 2,3-Diamino-4,6-dim~;lhyl~ylidine - Following the procedure of Example 45(a), except substituting 2-amino-4,6-dimethyl-3-nitl~ylidine (1.0650 g, 6.37 mmol) for the 4-amino-3-niLluben~otrifluoride, the title compound (836.1 mg, 95.7%) was prepared. This was used without char~ct~ri7~tion.
c) 4-Aza-2-[N-(benzyloxycarbonyl)-N-methyl]aminomethyl-5,7-dimethylbenzimidazole Following the procedure of Example 45(b), except sub~ 2,3-diamino-4,6-dhllt;lllylpyridine (836.1 mg, 6.09 mmol) for the 3,4-diaminobenzotrifluoride, the title compound (1.2273 g, 62%) following silica gel chromatography (3%
MeOH/CHCl3): MS (ES) m/e 325.0 (M~H)+.
d) 4-Aza-2-(methylaminomethyl)-5,7-dimethylben7imitl:~7ole Following the procedure of Example 45(c), except substituting 4-aza-2-[N-(benzyloxycarbonyl)-N-methyllarninomethyl-5,7-dimethylben7imi~1~7.ole (1.2273 g,3.78 mmol) for the 2-[N-(benzyloxycarbonyl)-N-methyl~aminomethyl-5-trifluoromethylbenzimidazole, the title compound was obtained as a white powder following trituration with Et2O. This material was used without characterization.
e) Methyl (S)-2,3,4,5-tetrahydro-7-[[[(4-aza-5,7-dimethylbe7imi~1~7c!1-2-yl)methyl]methylamino~carbonyl~-4-methyl-3-oxo- 1 H- 1,4-benzodiazepine-2-acetate Methyl (S)-7-carboxy-4-methyl-3-oxo-2,3,4,5-tetrahydro- lH- 1,4-benzodiazepine-2-acetate (175.0 mg, 0.60 mmol) was weighed into a 100 mL round bottomed flask. CH3CN (10 mL) was added, followed sequentially, by HOBt ~ H~O
(115.9 mg, 0.86 mmol), EDC (124.9 mg, 0.65 mmol), and diisopropylethylamine (0.13 mL, 0.75 mmol). A ~uspel sion of 4-aza-2-(methylaminomethyl)-5,7-dimethylben7imicl~7~-1e (144.5 mg, 0.76 mmol) and diisopropylethylamine (0.13 mL, 0.75 mmol) in CH3CN was added, and the reaction was stirred at RT. After 22 - 35 hr, the reaction was evaporated under V~l~;UUlll, and the residue was co-evaporated with toluene. Silica gel chromatography (3% MeOH/CHCl3 (1 L) then 5%
W O 97/24119 PCTrUS96120748 MeOH/CHCI3) gave the title compound (76.9 mg, 28%): MS (ES) m/e 465.2 (M+H)+.
f) (S)-2,3,4,5-Tetrahydro-7-[[[(4-aza-5,7-dimethylben7imi~ 7OI-2-yl)methyl]methylarnino]carbonyl-4-methyl-3-oxo-lH-1,4-benzodiazepine-2-acetic acid Methyl (S)-2,3,4,5-tetrahydro-7-[[[(4-a_a-5,7-dimethylbç7imi~7ol-2-yl)methyl]methylamino]carbonyl]-4-methyl-3-oxo- 1 H- 1,4-benzodiazepine-2-acetate (76.9 mg, 0.17 mmol) was dissolved in MeOH (5 mL) and HlO (5 rnL), and 1.0 N
- 10 NaOH (0.5 mL, 0.5 mmol) was added. After 24 hours at RT, the reaction was neutralized with 1.0 N HCl (0.5 mL), and the solvents were evaporated under vacuum. ODS chromatography (gradient: 5% CH3CN/H20 cont"inin~ 0.1% TFA
(500 mL), then 10% % CH3CN/H20 cont"ining 0.1 % TFA (500 mT~), then 15% %
CH3CN/H~0 cont"ining 0.1% TFA (500 mL), then 30% % CH3CN/EI~0 cont~ining 0.1 % TFA (500 mL)) gave a residue which was co-evaporated once with toluene anddried under high vacuum. The resulting residue was dissolved in MeOH (5 mL) and precipitated with Et,O. The white solid was collected on a sintered glass funnel and dried in a vacuum desiccator overnight to yield the title compound (52.0 mg, 68%):
HPLC (ODS column, 1.5 mL/rnin; gradient 5-50% CH3CN/H20 cont~ining 0.1%
TFA) tR 12.38 min; MS (ES) m/e 451.2 (M+H) ' . Anal Calcd for Cr,H~6N604 1 H20 1 CF3CO2H: C, 51.55; H, 5.02; N, 14.43: Found: C, 51.34; H, 5.00; N, 14.41.
Fx~ml?le 4g ~l~p~dtion of (S)-2.3.4.5-tetrahydro-7-rrr(5.6-difluoroben7imi~1~7ol-2-yl)methyllmethylaminolcarbonyll-4-methyl-3-oxo- 1 H- 1.4-benzodiazepine-2-açeticacid a) 1,2-Diarnino-4,5-difluorobenzene Following the procedure of Example 45(a), except substit-lting 4,5-difluoro-2-nitroaniline (2.0 g, 11.49 mmol) for the 4-amino-3-nitTobellzol~ ori~ , the title compound was prepared. This was used without characterization.
b) 2-[N-(Benzyl-~yca~bonyl)-N-methyl~aminomethyl-5,6-difluorobel-7i . . . i~ 7ole Following the procedure of Example 47(b), except substituting 1,2-~ mino-4,:~-difluoro~en~ne. for the 1,2~ mino-3-methylbenzene, and running the AcOH
cyclization step at 80~C instead of at 110~C, the title compound (1.3767 g, 36%) was W O 97/2411g PCTrUS96/20748 prepared: TLC Rf (1: 1 CH2C12/Et2O) 0.42; MS (ES) m/e 332.0 (M+H)+; IH
NMR (250 MHz, CDC13) ~ 7.50-7.14 (m, 8H), 5.13 (s, 2H), 4.61 (s, 2H), 3.06 (s, 3H).
S c) 5,6-Difluoro-2-(methylaminomethyl)ben7imid~7Ole Following the procedure of Example 46(b), except substituting 2-[N-- (benzyloxycarbonyl)-N-methyl]aminomethyl-5,6-difluorobçn7imi-1~7ole (1.3767 g, 4.16 mmol) for the 2-[N-(benzyloxycarbonyl)-N-methyl]~minom~thyl-4,7-~lim~thoxybe~ mdazole, the title compound (875.6 mg, qll~ntit~tive) was prepared:
10 MS (ES) m/e 198.0 (M+H)'.
d) Methyl-(S)-2,3,4,5-tetrahydro-7-[[[5,6-difluoroben7imill~7ol-2-yl)methyl]methylamino]carbonyl]-4-methyl-3-oxo- 1 H- 1,4-benzodiazepine-2-acetate Methyl (S)-7-carboxy-4-methyl-3-oxo-2,3,4,5-tetrahydro-lH-1,4-15 benzodiazepine-2-acetate (415.7 mg, 1.42 mmol) was taken up in CH3CN, and HOBt ~ H2O (209.3 mg, 1.55 mmol) and EDC (314.9 mg, 1.64 mmol) were added. After 5 min, and diisopropylethylamine (0.25 mL, 1.64 mmol) was added, which produced a clear, colorless solution. A solution of 5,6-difluoro-2-(methylaminomethyl)ben7imicl~701e (284.5 mg, 1.44 rnmol) in CH3CN was added.
20 After 30 minutes, the reaction became slightly turbid, so more diisopropylethylamine (0.25 mL) was added, which made the reaction again clear and colorless. After 24 hr, the reaction was evaporated under vacuum. The residue was co-evaporated once with toluene, then was chromatographed on silica gel (CHCl3 (0.25 L), then 2% MeOH/CHCl3 (1.5 L), then 5% MeOH/CHCl3) to afford 25 the title compound (456.8 mg, 68%): MS (ES) m/e 472.2 (M+H)t; 'H NMR (250 MHz, CDCl3) o 7.34-7.08 (m, 6H), 6.44 (br s, lH), 5.39 (d, J = 16.2 Hz, lH), 3.70 (s, 3H), 3.14 (s, 3H), 2.96 (s, 3H).
e) (S)-2,3,4,5-Tetrahydro-7-[[[(5,6-difluorobçr 7imi~ 7ol-2-30 yl)methyl]methylamino]carbonyl]-4-methyl-3-oxo- lH- 1,4-benzodiazepine-2-acetic acid Methyl (S)-2,3,4,5-tetrahydro-7-[[[5,6-difluoroben7imi~1~7-1-2-yl)methyl]methylamino]carbonyl] -4-methyl-3-oxo- l H- 1,4-ben70~ 7~pine-2-acetate (456.8 mg, 0.97 mmol) was dissolved in MeOH (10 mL) and H2O (10 ml). 1.0 N
35 NaOH (3.0 mL, 3.0 mmol) was added and the reaction was stirred at RT. After 18 hr, the reaction was neutralized with 1.0 N HCl (3.0 mT .). A white precipitate formed which was collected on a sintered glass filter and dried in a vacuum W O 97~4119 PCTrUS96/20748 desiccator. ODS chromatography (gradient: 10% CH3CN/H20 Cont~ining 0.1% TFA (500 rnL), then 18% % CH3CN/H20 cont~inin~ 0.1% TFA (500 mL), then 25% % CH3CN/H20 cont~inin~ 0.1% TFA (500 mL)) gave a residue which was co-evaporated once with toluene. The rçslllting residue was dissolved in a small 5 amount of MeOH and precipitated with Et2O to give the title co~ oulld (330.9 mg) as a white powder: HPLC (ODS column; 1.5 mL/min; gradient 5-50% CH3CN/H~O
contzlining 0.1% TFA) tR = 14.12 min; MS (ES) m/e 458.2 ~M+H)~. Anal Calcd for C22H2,N504F2 ~ 2.5 H2O: C, 52.57; H, 5.22; N, 13.94: Found: C, 52.76; H, 5.15; N, 67.
F.x~m~ple 50 Preparation of (S)-2.3.4.5-tetrahydro-7-rr~(4-:~7~-5-methylbe1l7i..lid~7.ol-2-yl)methylJamirlolcarbonyll-4-m~o-tllyl-3-oxo-lH-l~4-benzodiazepine-2-acetic acid a) Methyl (S)-2,3,4,5-tetrahydro-7-[[~(4-aza-5-methylben7imi-1~7nl-2-yl)methyl]amino]carbonylJ-4-methyl-3-oxo- lH- 1,4-benzodiazepine-2-acetate Methyl (S)-7-carboxy-4-methyl-3-oxo-2,3,4,5-tetrahydro- 1 H- 1,4- -benzodiazepine-2-acetate (228.8 mg, 0.78 mmol) was taken up in CH3CN, and HOBt ~ H~O (154.2 mg, 1.14 mmol), EDC (179.4 mg, 0.94 mmol), and diisopropylethylamine (0.50 ml~ ,, 0.94 rnmol) were added sequentially. A solution of 2-(aminomethyl)-4-aza-5-methylben7imi(1~7(1e dihydrochloride (125.4 mg, 0.77 mmol) in CH3CN /DMF was added, and the reaction was stirred at RT. After 24 hr, the reaction was evaporated under vacuum~ and the residue was co-evaporated oncewith toluene. Silica gel chromatography (CHCl3 (0.25 L) then 3% MeOH/CHCl3 (0.5 L), then 5% MeOH/CHCl3) gave the title compound (159.9 mg, 48%): MS (ES) m/e 437.2 (M+H)+.
b) (S)-2,3,4~5-Tetrahydro-7-[~[(4-aza-5-methylbenzimidazol-2-yl)methyl]amino]carbonyl]-4-methyl-3-oxo- lH- 1,4-benzodia_epine-2-acetic acid Following the procedure of Fx~mrle 48(f), methyl (S)-2,3,4,5-tetrahydro-7-[[[(4-aza-5-methyl~en7imi~ 7Ol-2-yl)methyl]aminoJcarbonyl]-4-methyl-3-oxo-lH-1,4-benzodiazepine-2-acetate (159.9 mg, 0.37 mmol) was saponified and purified to afford the title compound: MS (ES) m/e 423.39 (M+H)~. Anal. Calcd for C2,H22N6O4 - 0.5 H2O 1.25 TFA: C, 52.64; H, 5.02; N, 16.74. Found: C, 52.65; H, 5.02; N, 16.74.
Fx~m,ple 51 Preparation of (S)-2.3.4.5-tetrahydro-4-m~tllyl-7-rrr(4-nitrobçn7imicla7ol-2-yl)methyl~methylaminolcarbonyll-3-oxo-lH-1.4-benzo~ 7~pine-2-acetic acid a) 2-[N-(tert-Butoxycarbonyl)-N-methyl]aminomethyl-4-nitroben7imi<1~7ole e Boc-sarcosine (2.0320 g, 10.74 mmol) was dissolved in dry THF and cooled in a dry-ice/acetone bath to -15~C. Triethylamine (5.0 mL, 3.6375 mmol) was added, followed by isobutylchlor~fullllate (1.5 mL, 11.56 mmol). After 0.5 hr, the mixture was added to a solution of 1,2-diamino-3-nitrobenzene (1.3047 g, 10.77 mmol) in dry THF at -20~C, and the reaction was allowed to warm to RT. After 24 hr, the reaction was evaporated under vacuum and the residue was partitioned between EtOAc and 1.0 N NaHCO3. The layers were separated, and the aqueous layer was extracted with EtOAc. The combined organics were dried (MgSO4), filtered, and evaporated under vacuum. The residue was dissolved in glacial AcOH(100 mL), and the solution was heated at 75~C. After 24 hr, the reaction was evaporated under vacuum, and the residue was co-evaporated with toluene (2 x).
The m~t~ri~l was adsorbed onto silica gel and flash chromatographed on a dry silica gel column (gradient: CHC13 (0.5 L), then 2% MeOH/CHCl3 (1 L), then 5%
MeOH/CHCl3) to afford the title compound (2.2089 g, 75%): 'H NMR (250 MHz, CDCl3) ~ 8.10 (dd, 2H), 7.40-7.32 (m, lH), 4.69 (s, 2H), 3.02 (s, 3H), 1.54 ~s, 9H).
b) 2-(Methylaminomethyl)-4-niliuben,i.l.i(1~7Ole 2-[N-(tert-Butoxycarbonyl)-N-methyl3aminomethyl-4-nitroben7.imi~ 7.ole (2.2089 g, 8.05 mmol) was treated with 4 N HCl in dioxane at RT. After the addition, there was an imm~ te precipitation of a white solid. After 4 hr, the reaction was evaporated under vacuum and the residue was triturated with diethylether to give the title compound (1.639 g) as a white solid. This was used without chara~ ation.
c) Methyl-(S)-2,3,4,5-tetrahydro-7-1[[(4-nitrobe~7.imi~1~7.ol-2-yl)methyl]methylamino~carbonyl-4-methyl-3-oxo- 1 H- 1,4-benzodiazepine-2-acetateFollowing the procedure of Example 49(d), except substitllting 2-(methylaminomethyl)4-nitroben7.imi~1~7c-1e for 5,6-difluoro-2-(methylaminomethyl)ben7imi-1~7ole, the title compound (292.9 mg, qn~ntit:~tive) was plep~d: MS(ES) m/e 481.2 (M~H)+.
CA 0224l633 l998-06-26 W O 97/24119 PCTrUS96/20748 d) (S)-2,3,4,5-Tetrahydro-7-t[[(4-nitro~çn7imitl~7ol-2-yl)methyl]methylamino]carbonyl-4-methyl-3-oxo- l H- l ,~bP,n7.o~ .pine-2-acetic acid Following the procedure of Example 45(e), methyl ~S)-2,3,4,5-tetrahydro-7-[[[(4-nitrobenzimidazol-2-yl)methyl]methylamino]carbonyl-4-methyl-3-oxo- 1 H- 1,4-benzodiazepine-2-acetate (292.9 mg, 0.61 mmol) was saponified to afford the title compound (211.0 mg, 68%): MS (ES) m/e 467.4 (M+H)+. Anal. Calcd for C22H22N6O6 2.5 H20: C, 52.12; H, 5.27; N, 16.58: Found: C, 52.07; H, 4.97; N, 16.40.
F.xzln~?le 52 .,.tion of (S)-2.3.4.5-tetrahydro-7-rrr(4-arninoben7imi-l~7ol-2-yl)m~thyllmethylaminolcarbonyl-4-methyl-3-oxo-lH- 1 ~4-benzodiazepine-2-acetic ~i~
a) (s)-2~3~4~5-tetrahydro-7-[[[(4-amino~rn7imi~ 7ol-2-yl)methyl] methylamino}carbonyl-4-methyl-3 -oxo- 1 H- 1,4-benzodiazepine-2-acetic acid (S)-2,3,4,5-Tetrahydro-7-[[[(4-nitroben7imir1~ol-2-yl)methyl]methylamino]carbonyl-4-methyl-3-oxo- 1 H- 1,4-benzodiazepine-2-acetic acid (108.7 mg, 0.21 mmol) was dissolved in MeOH, and a catalytic amount of 10%
PdlC was added. The reaction was purged with Hz, then was stirred at RT under H2(balloon). After 20 hr, the catalyst was removed by filtration through celite(g), and the filtrate was evaporated under vacuum. The resulting solid was dissolved in MeOH, reprecipitated with Et20, dried in a vacuum rlecicc;~tor, and purified by ODS
chromatography (gradient: H2O cont~inin~ 0.1% TFA (500 mL), then 5%
CH3CN/H20 cont~ining 0.1% TFA (500 mL), then 10% % CH3CN/H70 Cont~inin~
0.1% TFA (500 mL), then 15% % CH3CN/H20 confz-inin~ 0.1% TFA (500 rnL), then 20% % CH3CN/H20 cont~inin~ 0.1% TFA (500 mL), then 25% % CE~3CN/Hz0 contz~ining 0.1% TFA (500 rnL), then 30% % CH3CNIH20 cont:~lininp: 0.1% TFA
(500 mL)). The rçsnltin~ m~tPri~l was co-evaporated once with toluene then was triturated with diethyl ether to afford the title compound (34.7 mg): MS (ES) m/e 437.5 (M+H)~; Anal. Calcd for C~24N6O4 1.5 H2O 1.5 TFA: C, 47.32; H, 4.53; N.
13.24: Found: C, 47.35, H, 4.86; N. 13.61.
Fx~m;l?le 53 W O 97/24119 PCTnUS96120748 ?ari.lion of 23.4~5-tetrahydro-7-rrr(lR~-fb~n7imi(1~7ol-2-yl)ethyllmethylarninolcarbonyll-4-m~thyl-3-oxo-lH-1.4-benzodia_epine-(2S~-aceticacid -a) 2-[1 (R)-~N-(Benzyloxycarbonyl)-N-methyl]aminoethyl~ben7.imifl~7.ole Following the procedure of Example 47(b), except ~ub~liluLillg Cbz-N-methyl-D-alanine for the Cbz-sarcosine, and substituting 1,2-phenylen~ mine for the 1,2-diamino-3-methylbenzene, and running the AcOH cyclization step at 80~C
10 instead of at 110~C, the title compound was prepared: MS (ES) m/e 310.2 (M+H)+.
b) 2-[1(R)-(Methylaminoethyl)]ben7.imi~ 7.01e Following the procedure of Example 46(b), except sub~ ulillg 2-[l(R)-tN-~benzyloxycarbonyl)-N-methyl]aminoethyl]ben7.imi(1~7.01e for the 2-[N-15 (benzyloxycarbonyl)-N-methyl]aminomethyl-4,7-dimethoxyben7imic~:~7.ole, the title compound (276.0 mg, 48%) was prepared: MS(ES) m/e 176.2 (M+H)+.
c) Methyl 2,3,4,5-tetrahydro-7-[[[(lR)-(ben7.imi~1~7.c!1-2-yl)ethyl]methylamino]ca~bollyl]-4-methyl-3-oxo- 1 H- 1,4-benzodiazepine-2-acetate Following the procedure of Example 49(d), except substituting 2-[l(R)-(methylaminoethyl)]benzimidazole for the 5,6-difluoro-2-(methylaminomethyl)1)~ll7.il"i(1~7.ole, the title compound (203.5 mg, 90%) was prepared: MS (ES) m/e 450.5 (M+H)~.
25 d) 2,3,4,5-Tetrahydro-7-[~[(lR)-(bçn7.imi-1~7.ol-2-yl)ethyl]methylamino]carbonyl]-4-methyl-3-oxo- 1 H- 1,4-benzodiazepine-2-acetic acid Following the procedure of Example 49(e), methyl 2,3,4,5-tetrahydro-7-[[[(lR)-(b~7imi~1~7.ol-2-yl)ethyl]methylamino~ca~bonyll-4-methyl-3-oxo-lH-1,4-benzodiazepine-2-acetate was saponified to afford the title compound (179.3 mg, 30 75%) following ODS chromatography: MS (ES) m/e 436 5 (M~H)+. Anal. Calcd for C23H25N504 0.75 H20 0.75 TFA: C, 55.01; H, 5.14; N, 13.10; Found: C, 54.98;
H, 5.42; N. 12.75.
F.x~n~le 54 CA 0224l633 l998-06-26 W O 97/24119 PCTrUS96/20748 Pl~alion of ethyl fS)-2.3~4.$-tetrahydro-7-rrr(4-Aza-5-met~lylben7.imid~ol-2-yl)methyllaminolcarbonyll4-metlly1-3-oxo- 1 H- 1.4-'~n70~ 7~pine-2-acetate S a) Ethyl (S)-2,3,4,5-tetrahydro-7-[[[(4-Aza-5-methylben7imi~1~701-2-yl)methyl]amino]carbonyl]4-methyl-3-oxo- lH- 1 .4-bt~n70tli~7~pine-2-acetate (S)-2,3,4,5-Tetrahydro-7-[[[(4-aza-S-methylbçn7.imi~7.oI-2-yl)methyl]arnino]carbonyl]-4-methyl-3-oxo- 1 H- 1 ,4-benzodiazepine-2-acetic acid ~0.5 g) was dissolved in EtOH, and the solution was cooled in an }ce bath to 0~C.
Gaseous HCI was bubbled into the solution until the solution was saturated, then the flask was sealed with a rubber septum and the cooling bath was removed. The reaction was stirred at RT for 20 hr, then the solvents were evaporated under vacuum. The residue was co-evaporated three times with toluene (3 x), then was dissolved in EtOH and precipitated with Et20. The solid was collected on a sintered glass funnel and dried in a vacuum ~iesiccatQr overnight to afford the title compound (483.9 mg): MS (ES) m/e 451.4 (M+H)~. Anal. Calcd for C23H26N6O, ~ HCl ~ 1.375 H20: C, 53.98; H, 5.86; N, 16.42. Found: C, 54.00; H, 5.82; N, 16.42.
F.xz-m~?le S5 PIG~al~Lion of 2.3.4.5-tetrahydro-7-rrr(lS)-(ben7imi-1~7.ol-2-yl)etl~yllmethyl~minolcarbonyll-4-methyl-3-oxo-lH-1 .4-benzodi~epine-~2S)-aceticacid a) 2-[l(S)-[N-(tert-Butoxycarbonyl~-N-methyl~aminoethyl]ben7imi~1~701e Following the procedure of Example 51(a) except substituting Boc-N-methyl-L-alanine for the Boc-sarcosine, and sub..liLuLillg 1,2-phenylene~ min~ for the 1,2-diamino-3-nitroben7~n~, the title compound (1.7792 g, 65%) was prepared following recryet:llli7~tion from CHCI3/hexanes: MS (ES) m/e 276.4 (M+H)~.
b) 2-[l(S)-(Methylaminoethyl)]ben7imi~1~7.ole Following the procedure of Example 51(b), except substituting 2-[l(S)-~N-(tert-butoxycal'L,ollyl)-N-methyl]aminoethyl]bel-7i . . .i-i~7Qle for the 2-[N-(tert-butoxycarbonyl)-N-methyl]aminomethyl4-nitroben7imi~ 701e, the title compound 35 was prepared. This was used without charactt-ri7~tion.
W O 97/24119 PCT~US96/20748 c) Methyl (S)-2,3,4,5-tetrahydro-7-[[t(1 S)-(ben7.imi~1~701-2-yl)ethyl]methylamino]carbonyl]-4-methyl-3-oxo- 1 H- 1,4-bçn7.o~ 7P-pine-2-acetate Following the procedure of Example 49(d), except substihlting 2-[l(S)-(methylaminoethyl)]ben7.imitl~7.(1e for the 5,6-difluoro-2-S (methylaminomethyl) ben7.imi~1~7.01e, the title compound (414.7 mg, 88%) was prepared: MS (ES) m/e 450.2 (M+H)+.
d) 2,3,4,5-Tetrahydro-7-t[[(lS)-(ben7.imi~ 7.~1-2-yl)ethyl]methylamino]carbonyl]-4-methyl-3-oxo- lH- 1,4-benzodiazepine-2-acetic acid 1(~ Following the procedure of Example 45(e), methyl (S)-2,3,4,5-tetrahydro-7-[[[(lS)-(ben7.imi~1~7.ol-2-yl)ethyl~methylamino]carbonyl]-4-methyl-3-oxo-lH-1,4-benzodiazepine-2-acetate was saponified to afford the title compound (117.2 mg):MS (ES) m/e 436.2 (M+H)+. Anal Calcd for C23H~5N504 0.75 H20 0.75 TFA: C, 55.05; H, 5.14; N, 13.10; Found: C, 55.14; H, 5.38; N, 13.04.
Ex~mple S6 P~ io~ of 2.3.4.5-tetrahydro-7-rrr(lS)-(b~n7imi~1~7.ol-2-yl)ethyllaminolcarbonyll-4-methyl-3-oxo-lH-1.4-ben7.o~;~7.~pine-(2S)-acetic acid a) 2-r 1 (S)-(tert-Butoxycarbonyl)aminoethyl]ben7imi~1~7.ole Following the procedure of Example 51 (a) except substituting Boc- L-alanine for the Boc-sarcosine, and sul)~LiLuLil~g 1,2-phenylene~ mine for the 1,2-diamino-3-nitroben7ene7 the title compound (714.7 mg, 25%) was prepared: MS
(ES) m/e 262.4 (M+H)+.
b) 2-[1 (S)-(Aminoethyl)]ben7imi(1~7.ole Following the procedure of Example 51(b), except subsliluli~lg 2-[l(S)-(tert-butoxycarbonyl)aminoethyl]ben7imi~1~701e for the 2-[N-(tert-butoxycarbonyl)-N-methyl]aminomethyl-4-nitroben7.imi~1~7.01e, the title compound was prepared. This was used without characterization.
c) Methyl 2,3,4,5-tetrahydro-7-[t[(1S)-(ben7imi~ 7.ol-2-yl)ethyl)]amino]carbonyl]-4-methyl-3 -oxo- l H- 1,4-ben 70~ 7epine-2-acetate Following the procedure of Example 49(d), except substit~ting 2-[l(S)-(aminoethyl)]ben7.imi~1~7.r 1e for 5,6-difluoro-2-(methylaminomethyl)be~- ~.i " ,icl~7.~1e, the title compound (270.7 mg, 80%) was p~ l.,d: MS (ES) m/e 436.0 (M+H~+.
W O 97/24119 PCT~US96120748 d) 2,3,4,5-Tetrahydro-7-t[[(lS)-(be~7imid~7ol-2-yl)ethyl)]amino]carbonyl]-4-methyl-3-oxo- lH-1,4-benzodiazepine-2-acetic acid Following the procedure of Example 45(e), methyl 2,3,4,5-tetrahydro-7-[[[(1 S)-(benzimidazol-2-yl)ethyl)]amino]carbonyl]-4-methyl-3-oxo- lH- 1,4-benzodia_epine-2-acetate was saponified to afford the title compound (158.1 mg, 61%): MS (ES) m/e 422.0 (M+H)+. Anal. Calcd for C2lH23N5O4 ~ 1.75 H20: C, 58.37; H, 5.90; N, 15.46; Found: C 58.17; H, 5.77; N, 15.08.
F.~z~ml?le 57 P~ lion of 2.3.4~5-tetr~l~ydro-7-rrr(lR~-(ben~ 7ol-2-yl)ethyllarninolcarbonyll-4-methyl-3-oxo-lH-1.4-benzodiazepine-(2S~-acetic acid a) 2-[1 (R)-(Benzyloxycarbonyl)aminoethyl]ben7imi~1~701e Following the procedure of Example 47(b~, except substituting Cbz-D-alanine for the Cbz-sarcosine, and substituting 1,2-phenylenedi~minc for the 1,2-diarnino-3-methylbenzene, and rurming the AcOH cyclization step at 80~C instead of at 110~C, the title compound (1.1455 g, 43%) was prèpared: MS (ES) m/e 296.4 (M+H) i .
b) 2-(l(R)-Aminoethyl)ben~imi-1~7O1e Following the procedure of Fx~mple 46(b), except sub~iLu~ g 2-[l(R)-(benzyloxycarbonyl)aminaethyl]ben7imi~ 701e for the 2-[N-(benzyloxycarbonyl)-N-methyl]aminomethyl-4,7-dimethoxyben~ 7Qle, the title compound (258.1 mg, 93%) was ~l~palt;d: MS(ES) m/e 161.9 (M+H)t.
c) Methyl 2,3,4,5-tetrahydro-7-[[[( lR)-(ben7imi~1~7~-1-2-yl)ethyl~amino]carbonyl]-4-methyl-3-oxo- 1 H- 1,4-benzodiazepine-(2S)-acetate Following the procedure of Example 49(d), except substituting 2-(l(R)-aminoethyl)ben7imi~701e for the 5,6-difluoro-2-(methylarninomethyl)benzimidazole, the title compound (263.6 mg, 84%) was prepared: MS (ES) m/e 436.3 (M+H)+.
d) 2,3,4,5-Tetrahydro-7-[[[(lR)-(be~7imi(i~7ol-2-yl)ethyl]amino]carbonyl]-4 methyl-3-oxo- lH- 1,4-ben7o~ 7topine-2-acetic acid W O 97/24119 PCTrUS96/20748 Following the procedure of F.xz-mple 49(e), methyl 2,3,4,5-tetrahydro-7-[[[(lR)-(b~n7imi~1~7ol-2-yl)ethyl]amino]carbonyl]~-methyl-3-oxo-lH-1,4-benzodiazepine-(2S)-acetate was saponified to afford the title compound (125.0 mg, 49%): MS (ES) m~e 422.0 (M+H)t. Anal. Calcd for C22H23N5O4 ~ 0.5 H20 ~ 1.25 HCl: C, 55.51; H, 5.35; N, 14.71. Found: C, 55.70; H, 5.47; N, 14.53.
- Example $8 Pr~a,dLion of (S)-2.3~4.5-tetrahydro-7-rrrCjmifl:~7o( 1 .2a)pyrid-2-yl)methyllmethylaminolcarbonyl~-4-methyl-3-oxo-lH-1.4-benzodiazepine-2-acetic acid a) 2-Carboethoxyirnidazo[ 1 ,2a]pyridine 2-Aminopyridine (4 g, 42.50 mmol) was dissolved in MeOH (50 mL). Ethyl bromopyruvate (8.3 g, 42.50 mmol) was added and the reaction was stirred at 70~Cfor 2 hr. The solvent was then elimin~te-l and the solution was neutralized with 1 M
NaOH. The reaction was extracted with EtOAc, and the combined EtOAc layers were washed with brine. Drying (MgSO4), filtration, concentration, and silica gel ilash column chromatography (2% MeOH/Cl2CH2) gave the title compound (4.5 g, 56%) as a pale yellow solid: 1H NMR (400 MHz, CDC13) o 1.43 (t, J = 7 Hz, 3H), 4.45(q,J=7Hz,2H),6.86(t,J=6.6Hz, lH),7.24(t,J=6.6Hz, lH).
b) 2-Hydroxymethylimicl~7O[1,2a]pyridine 2-Carboethoxyimidazo[1,2a]pyridine (0.5 g, 2.81 mmol) was dissolved in dry THF at 0~C, and then a solution of lithium ~ minl~m hydride (0.5 mL of a 1.0M in THF) was added. The reaction was allowed to warm to RT and stirred for 1 hr.
H2O (0.2 rnL) was added, followed by 15% NaOH (0.2 mL), and finally H2O (0.6 mL). The solids were removed by filtration and washed with hot THF (2 x 100 mL) and hot CHCl3 (4 x 100 mL). The filtrate and washings were combined and dried (MgSO4). Following filtration the solvents were removed under reduce pressure the residue was purified by silica gel flash column chromatography (5%
MeOH/Cl2CH2~ to obtain the title co~ u-ld (0.1 g, 25~o) as pale yellow liquid: 'H
NMR (400 MHz, CDCl3) o 4.85 (s, 2H), 6.76 (t, J = 6.8 Hz, lH), 7.15 (t, J = 6.8 Hz, lH), 7.53 (s, lH), 7.54 (d, J = 6.7 Hz, lH), 8.1 (d, J = 6.7 Hz, lH). MS (ES) m/e 149 (M + H) +.
,.
c) 2-Chloromethylimi~1~7o[ 1 ,2a]pyridine CA 0224l633 l998-06-26 W O 97/24119 PCTnjS96nO748 Thionyl chloride (0.4 mL, 3.2 mmol) was added to a solution of 2-hydroxymethylimi(l~70[1,2a]pyridine (0.4 g, 2.7 mmol) in CHC13 (30 mL) at 0~C
After stirring at RT for 1 hr, the suspension was poured into a mixture of ice, 10%
NaHCO3, and CHCl3. The layers were separated, and the aqueous phase was extracted with CHCI3. The organic extracts were combined and dried (MgS04).
Following filtration, the solvents were removed under reduce pressure to obtain the title compound (0.4 g, 89%). lH NMR (400 MHz, CDCl3) o 4.7 (s, 2H), 6.7 (t, lH~, -7.2 (t, lH), 7.5 (d, lH), 7.6 (s, lH), 8.0 (d, lH); MS (ES) m/e 167 (M + H)+.
d) 2-(Methylaminomethyl)imidazo[1,2a]pyridine Freshly condensed methylamine (15 mL) was added to a solution of 2-chloromethylimifl~70~1,2a]pyridine (317 mg, 2 mmol) in EtOH (5 mL) at 0~C, and the reaction mixture was allowed to stir at 0~C for 2 h. The solvent was then ~limin~tto~l and the residue was purified by reverse-phase column chromatography(C-18 silica gel, H20 cont~ining 0.1 % TFA). Lyophilization gave the title compound (461 mg, 87%) as a white solid: IH NMR (250 MHz, CDC13) o 2.5 (s, 3H), 4.0 (s, 2H), 6.7 (t, lH), 7.2 (t, lH), 7.5 (d, l~I), 7.6 (s, lH), 8.1 (d, lH). MS
(ES) m/e 162 (M + H)'.
- 20 e) Methyl (S)-2,3,4,5-tetrahydro-7-[[[(imidazo(1,2a)pyrid-2-yl)methyl]methylamino~carbonyl]-4-methyl-3-oxo- 1 H- 1,4-benzodiazepine-2-acetate Methyl (S)-7-carboxy4-methyl-3-oxo-2,3,4,5-tetrahydro- lH- I ,~
benzodiazepine-2-acetate (828.2 mg, 2.83 mmol) was taken up in CH3CN, and HOBt ~ H20 (398.1 mg, 2.92 mmol), EDC (562.9 mg, 2.94 mmol), and diisopropylethylamine (1 mL, 5.74 mmol) were added sequentially. When the solids had dissolved, a solution of 2-(methylaminomethyl)imi~ 70[1,2a]pyridine (460.7 mg, 2.86 mmol) and diisopropylethylamine (1.5 mL, 8.61 mmol) in CH3CN
was added, and the reaction was stirred at RT. After 24 hr, the reaction was concentrated, and the residue was co-~v~olat~d with toluene (2 x). The rçsl-lting residue was chromatographed on silica gel (5% MeOH/CHCl3) to give the title compound (694.6 mg, 56%): MS (ES) m/e 436.4 (M~H)~.
f) (S)-2,3,4,5-Tetrahydro-7-[[[(imi-1~7.o(1,2a)pyrid-2-yl)methyl]methylamino~carbonyl]-4-methyl-3-oxo- 1 H- 1,4-benzodiazepine-2-aceticacid Methyl (S)-2,3,4,5-tetrahydro-7-~[[(imidazo(1,2a)pyrid-2-yl)methyl]methylamino]carbonyl]-4-methyl-3-oxo- lH- 1,4-benzodiazepine-2-acetate 182 "
CA 0224l633 l998-06-26 W O 97/24119 PCT~US96/20748 (663.4 mg, 1.52 mmol) was dissolved in MeOH (10 mL). H2O (10 mL) was added, followed by 1.0 N NaOH (5 mL, 5.0 mmol), and the reaction was stirredat RT. After 20 hr, the reaction was neutralized with 1.0 N HCl (5 mL), and the solution was evaporated under vacuum. The residue was purified by ODS
chromatography (gradient: H2O containing 0.1% TFA (500 mL), then 5%
CH3CN/H20 con~aining 0.1% TFA (500 mL), then 10% CH3CN/H20 co~
- 0.1% TFA (500 mL), then 15% CH3CN/H2O cont~ining 0.1% TFA (500 mL), then 20% CH3CN/H2O cont~ining 0.1% TFA (500 rnL)) followed by rechromatography on ODS (gradient: H20 cont~inin~ 0.1~TFA (250 mL), then 10% CH3CN/H2O
cont~ining 0.1% TFA (1.5 L), then 20% CH3CN/H20 c~-nt~ininp 0.1% TFA (1 L).
Fractions containing pure m~t~ri~l were combined and concentrated. The residue was co-evaporated with toluene, then was dissolved in MeOH and reprecipitated with l~t20 to afford the title compound (96.4 mg): MS (ES) m/e 421.9 (M+H)~.
Anal. Calcd for C22H23N5O4 ~ 0.25 H2O ~ TFA: C, 53.38; H, 4.57; N, 12.97. Found:C, 53.68; H, 4.97; N, 12.94.
E~xample 59 Preparation of (+)-7-rrr(4~5-Dimethyl-lH-;mi~ ol-2-yl)methyllmethylaminolcarbonyll-2~3.4.5-tetrahydro-4-methyl-3-oxo-lH-1~4-benzodiazepine-2-acetic acid a) N-(Carbobenzyloxy)-N-(methyl)acetonitrile Cbz chloride (7.40 rnL, 49.3 mmol) was added slowly at RT to a solution of N methylaminoacetonitrile hydrochloride (5.0 g, 46.92 mmol) and triethylamine (13.4 mL, 96.2 mmol) in dichloromethane (200 mL). The reaction was stirred at RT18 h, and the mixture was washed with lN HCl, water and brine. The organic layerwas dried (MgSO4~ and concentrated to yield the title compound (6.97 g, 73%) as a clear oil.
b) N-(Carbobenzyloxy)-N-(methyl)aminothio~ret~mi~le Hydrogen sulfide was bubbled through a solution of N-(carbobenzyloxy)-N-(methyl)acetonitrile (15 g, 73.5 mmol) and triethylarnine (30.75 mL, 220.6 mmol) in DMF (250 mL). After 20 min, the flask was closed and the reaction was stirred atRT for 18 h. The reaction was then poured into 2 N NaHCO3 (1 L ) and extracted with dichloromPth~ne. The combined organic phase was washed with 1: 1 .
W O 97/24119 PCT~US96t20748 water/brine (5 x), dried (MgSO4), and concentrated to give a yellow oil which was purified by silica gel llash chromatography (step gradient, 40-50% ethyl acetate/hexane) to yield the title compound (12.26 g, 70%) as a white solid: MS
(ES) rn/e 239.0 [M+H]+.
s c) N-(Carbobenzyloxy)-N-methyl-S-(methyl)acetothioimidate Iodomt-.th~nP (17.66 mL, 283.6 mmol) was added at RT to a solution of N-(carbobenzyloxy)-N-(methyl)aminothio~et:lmide(6.75 g, 28.36 mmol) in acetone 100 mL). The solution was stirred at RT in the dark for 3 h, and the reslllting precipitate was filtered to yield the title compound (9.63 g, 89%) as a white solid:
MS (ES) m~e 253.4 [M+H]+.
d) (+)-2-Amino-3,3-dimethoxybutane Sodium cyanoborohydride was added to a solution of 3,3-dimethoxy-2-butanone (1.32 g, 10 mmol) and ammonium acetate (1.1 g, 100 mmol) in methanol (30 rnL). The pH was adjusted to 6 with methanolic HCl, and the reaction was stirred at RT overnight and concentrated. The residue was dissolved in water, and the pH was adjusted to 5 with aqueous HCl. The res-lltin~ solution was extractedwit_ ether (3 x), and the aqueous phase was basified to pH 10 with Na2CO3, and extracted with ether. The organic layer was dried (MgSO4) and concentrated to yield the title compound (1.1 g, 83%) as a clear oil: MS (ES) m/el34.2 [M+Hl+.
e) N-(Carbobenzyloxy)-N-methyl-(4,5-dimethyl- lH-imidazol-2-yl)meth~n~rnine A solution of (~)-2-amino-3,3-~l;m~tho~y~uLalle (1.05 g, 7.89 mmol) and N-(carbobenzyloxy)-N-methyl-S-(methyl)~< etofhi~ imidate (2.0 g, 5.26 mmol) in methanol (30 mL) was warmed at 60~C for 2 h, and concentrated to give a yellow oil. The crude oil was dissolved in 6 N HCl (30 mL) and stirred at RT 1 h. The solution was basified to pH 12 wieh aqueous NaOH and then extracted with dichlor~)m~th~ne. The combined organic phase was dried (MgSO4) and concentrated to give a brown oil which was purified by silica gel flash chromatography (4% methanol/dichloromethane) to yield the title compound (0.590 g, 41%) as a clear oil: MS (ES) m/e 274.0 [M+H~+.
f~ N-Methyl-(4,5-dimethyl-lH-imi~ 7,t~1-2-yl)~ nzlrninr.
A solution of N-(carbobenzyloxy)-N-methyl-(4,5-dimethyl-lH-imidazol-2-yl)mto.thz-nz~mine (0.35 g, 1.28 mmol) in methanol (15 mL) and glacial acetic acid (5 CA 0224l633 l998-06-26 W O 97/24119 PCTrUS96/20748 mL), cont~ining 10% Pd/C (0.035 g), was shaken in a H2 atmosphere (45 Psi) for 6 h. The reaction was filtered and filtrate conce~ dl~;d to give the title compound (0.22 g, 86%) as a brown oil which was used in the next step without further purification: MS (ES) m/e 140 [M+H]+.
g) Methyl (_)-7-[[[(4,5-dimethyl-lH-imi-1~7ol-2-yl)methyl]methylamino]carbonyl]-- 2,3,4,5-tetrahydro-4-methyl-3-oxo- 1 H- 1,4-benzodiazepine-2-acetate A solution of N-methyl-(4,5-dimethyl-lH-imi(1~7ol-2-yl)meth~n~minP (0.20 g, 1.1 mmol) and methyl (_)-7-carboxy-2,3,4,5-tetrahydro-4-methyl-3-oxo-1~-1,4-benzodia_epine-2-acetate (0.320 g, 1.1 mmol) in the presence of DIEA (0.287 ml, 1.65 mmol) was stirred at RT. EDC (0.316 g, 1.65 mmol) was then added, followed by DMAP (0.013 g, 0.11 mmol). The mixture was stirred at RT for 18 h, and concentrated to give an oil which was purified by silica gel flash chromatography (step gradient, 0.5-2% methanol/dichloromPthzlne) to yield the title compound (0.060 g, 9%) as a clear oil: MS (ES) m/e 414.2 [M+H]+.
h) (+)-7-[[[(4,5-Dimethyl-lH-imidazol-2-yl)methyl]methylamino]carbonyl]-2,3,4,5-tetrahydro-4-methyl-3-oxo- lH- 1,4-benzodiazepine-2-acetic acid 1 N Sodium hydroxide (3 eq) was added to a solution of methyl (+)-7-[[[(4,5-dimethyl-lH-imidazol-2-yl)methyl]methylamino]carbonyl]-2,3,4,5-tetrahydro-4-methyl-3-oxo-lH-1,4-benzodiazepine-2-acetate (0.060g, 0.145 mmol), the mixture was stirred at RT for 5 h, and concentrated. The residue was dissolved in water and the pH was adjusted to S with 50% acetic acid. The solution was concentrated andpurified by MPLC (ODS-AQ, 10% acetonitrile/water cont~inin~ 0.1% TFA, UV
detection at 220 nm) to yield the title compound (0.050 g, 86%) as a white solid:
MS (ES) m/e 400.2 [M+H]+.
Example 60 Preparation of 3-rr3-r2-(ben7imill~7- 1-2-yl)ethyllisoxazolin-SfR.S)-yllacetyllamino-3(R.S)-meth~ upalloic acid a) 4-(Ben7imi-1~7Ol-2-yl)- 1 -butene According to the general procedures of Preparation 4 in P50256-1, except ~ul~Li~ulillg 4-pentenoic acid for the Boc-sarcosine, the title compound is prepared.
-b) 4-( l -Toluenesulfonylben7imi~l~7Ql-2-yl~- l -butene W O 97124119 PCTrUS96/20748 Sodium hydride is added carefully to a solution of 4-(bP.n7.imi(1~7.01-2-yl)-1-butene (50 mmole) and 4-toluenesulfonyl chloride (55 mmole) in dry THF
(2~)0 mr .). The reaction is stirred at RT until complete, then is quenched withsaturated NH,,CI (200 mL~, and the mixture is extracted with EtOAc. The combined5 organic extracts are dried (MgSO4) and concentrated, and the residue is purified by silica gel chromatography to give the title compound.
-c) 4-(l-Toluenesulfonylben7.imi-1~7.ol-2-yl)-l-butanal Ozone is bubbled into a solutiolI of 4-(1-toluenesulfonylben7imi~l~7.Ql-2-yl)-l-butene (40 mmole) in CH2Cl~ (160 mL) and MeOH ~40 mL) at -78~C until the blue color persists, then the excess ozone is removed by bubbling argon through the solution. Dry dimethylsulfide (excess) is added, and the reaction is warmed to RT.
The reaction is stirred at RT until complete, then is concentrated, and the residue is chromatographed on silica gel to afford the title c~ oulld.
d) 4-(l-Toluenesulfonylben7.imi~1~7.(-1-2-yl)-l-butanal oxime Hydroxylamine hydrochloride (33 rnmole) is added to a solution of 4-(1-toluenesulfonylben7imi~1~7.ol-2-yl)-1-butanal (30 mmole) and anhydrous sodium acetate ~66 mmole) in MeOH (150 mL) at 0~C. The reaction is stirred at 0~C until20 complete, then is concentrated, and the residue is partitioned between H20 and EtOAc. The layers are separated, and the aqueous layer is extracted with EtOAc.
The combined organic layers are washed sequentially with 5% NaHCO3 and saturated brine, dried (MgSO4~, and concentrated to afford the title compound.
2~ e) 4-(l-Toluenesulfonylbenzimi~ 7.ol-2-yl)-l-butanoxirninoyl chloride According to the procedure of Example l(b) in WO 95/14682, except substituting 4-(1-toluenesulfonylbe~7.imifl~7.01-2-yl)-l-butanal oxime for the 4-cyanobenzoxime, the title compound is prepared.
f) tert-Butyl ~3-[2-(1 -toll-~neslllfonylbPn7imi(1~7ol-2-yl)ethyl]isoxazolin-5(R~S)-yl~acetate According to the procedure of Example 1 (d) of WO 95/14682, except sub.~ uling 4-(1-toluenesu~nylben7.imi<~ Ql-2-yl)-1-butanoximinoyl chloride for the 4-cyanobenzoximinoy~ chloride, and ~ub~liluling tert-butyl 3-butenoate for the methyl 3-butenoate, the title compound is ~l~al~d.
W O 97/24119 PCT~US96/20748 g) [3-[2-(1-Tnllleneslllfonylbenzimidazol-2-yl)ethyl]isoxazolin-S(R,S)-yl]acetic acid 4 M HCl in dioxane (10 mL) is added to a solution of tert-butyl [3-[2-(1-toluenesulfonylben7imi(1~7nl-2-yl)ethyl]isoxazolin-5~R,S)-yl]acetate (5 mmole) in C~I2CH2 (40 mL) at 0~C. The reaction is stirred at RT until complete, then is concentrated to afford the title compound.
h) Ethyl 3-[[3-[2-(1-toluenesulfonylben7imi-1~7ol-2-yl)ethyl]isoxazolin-5(R,S)-yl]acetyl]amino-3(R,S)-methylpropanoate EDC (1.2 mmole) is added to a solution of [3-[2-(1-toluenesulfonylbenzimidazol-2-yl)ethyl]isoxazolin-5(R,S)-yl]acetic acid (1 mmole), ethyl 3(R,S)-aminobutyrate (1.2 mmole), HOBt H2O (1.2 mmole), and diisopropylethylamine (4 mmole) in anhydrous CH3CN (5 rnL) at RT. The reaction is stirred at RT until complete, then is concentrated, and the residue is purified by silica gel chromatography to afford the title compound.
i) 3-[[3-[2-(Ben7imi-1~7ol-2-yl)ethyl]isoxazolin-5(R,S)-yl]acetyl]amino-3(R,S)-methylpropanoic acid 1.0 N LiOH (2.5 mmole) is added to a solution of ethyl 3-[3-[2-(1-toluenesulfonylb~ n7imi~1~7ol-2-yl)ethyl]isoxazolin-5(R,S)-yl]acetyl]amino-3(R,S)-methylpropanoate (0.5 mmole) in THF (2.5 ml). The reaction is stirred at RT until complete, then is neutralized with l.0 N HCl. The solution is concentrated and the residue is purified by reverse-phase chromatography to afford the title compound.
Fxample 61 Preparation of 3-{3.4-dihydro-8-rrr(ben7imi(1~7nl-2-yl)methyl~methy}aminolcarbonyll-1-methyl-2~5-dioxo-lH-1.4-benzodiazepine}-4-propanoic acid a) 2-Amino-4-iodobenzoic acid The title compound is prepared from the oxidation of 4-iodo-2-nitrotoluene to give 4-iodo-2-niLI~bell~oic acid according to the method of Sasson, et al., J. Org.
Chem. 1986, 51, 2880-2883, followed by reduction of the nitro group using iron and acetic acid.
W O 97/24119 PCTrUS96/20748 b) 4-Iodoisatoic anhydride To a mechanically stirred ice-cold solution of 2-amino-4-iodobenzoic acid (26.3 g, 0.1 mol), sodium carbonate ~10.6 g, 0.1 mol) and water (250 mL), is slowly added, via an addition funnel, a solution of phosgene in toluene (80 mL of 1.93 M
S solution). After 2 h, the precipitated product is i.~ol~te~l by filtration and the solid is washed cuccessively with water (200 mL), a 1: 1 mixture of ethanol/ether (300 mL),and ether (200 mL). Drying under vacuum yields the title compound.
c) N-(2-Amino-4-iodobenzoyl)-,13-alanine benzyl ester A magnetically stirred solution of 4-iodoisatoic anhydride (5.0 g, 0.0173 mol)"1~ alanine benzyl ester tosylate (5.85 g, 0.0173 mol), and dimethylaminopyridine (0.5 g, 0.0041 mol) in pyridine (35 rnL) is heated for 2 h at 80~C. The reaction mixture is allowed to cool to RT and concentrated in vacuo.
The resulting residue is dissolved in ethyl acetate (100 mL), and washed successively with 10% cupric sulfate (2 x 50 rnL), saturated sodium bicarbonate (I x 50 mL) and brine (1 x 50 mL). Drying (Na2SO4), filtration, concentration, and silica gel chromatography (1:1 EtOAc/hex~n~ s) gives the title compound.
d) N-(4-Iodo-2-methylaminobenzoyl)-,13-alar~ine benzyl ester A m~gnç~ lly stirred solution of N-(2-amino-4-iodobenzoyl)-13-alanine benzyl ester (2.0 mmol), 2,6-lutidine (0.35 mL, 3.0 mmol) and methyl iodide (0.19 mL, 3.0 mmol) in DMF (15 mL) is heated at 50~C for 15 h. The reaction mixture isallowed to cool to RT and concentrated in vacuo. The r~s-llting residue is dissolved in ethyl acetate (75 rnL), and washed succl ccively with 10% citric acid (1 x 50 mL), saturated sodium bicarbonate (1 x 50 mL) and brine (I x 50 mL). Drying (Na2SO4),filtration, concentration, and silica gel chromatography (gradient 35-65 %
EtOAc/hexanes) gives the title compound.
e) Benzyl 3-~3,4-dihyro-8-iodo- 1-methyl-2,5-dioxo- lH- 1,4-benzodiazepinel-4-propanoate To a cold (-30~C) m~gn~ti~:~lly stirred solution of N-(4-iodo-2-methylarninobenzoyl)-,13-alanine benzyl ester (0.305 g, 0.69 mrnol) and triethylamine (0.144 g, 1.04 mmol) in methylene chloride (3 mL) is added slowly a solution of a-bromoacetyl bromide (0.09 mL, 1.04 mmol) in methylene chloride (2 mL) under argon. The reaction mixture is allowed to warm to RT and stir for 2 h. The llli~lulG
is diluted with methylene chloride (40 ml ) and washed s-lcce~ivGly with 10% citric acid (1 x 50 rnL) and saturated sodium bicar~onate (1 x 50 mL), dried (Na2SO4), W O 97/24119 PCT~US96/20748 filtered and concentrated in vacuo. The resultin~ residue is dissolved in DMF (3 ml) and added via an addition funnel to a slurry of sodium hydride (25 mg, 1.04 mrnol) in DMF (2 mL) which is cooled to 0~C. After 2 h of stirring, the mixture is poured over an ice cooled solution of 10% citric acid (50 mL) and 5 extracted with ethyl acetate (3 x 40 ml). The combined extracts are washed with saturated sodium bicarbonate (1 x 50 mL), dried (Na2SO4), filtered and concentrated.
Silica gel chromatography (gradient 40-70% EtOAc/hexanes) gives the title compound.
f) Benzyl-3-[3,4-dihyro-8-t[[(ben7imi(1~7Ol-2-yl)methyl]methylamino]carbonyl]
methyl-2,5-dioxo- 1 H- 1,4-benzodiazepine]-4-propanoate A mixture of benzyl 3-[3,4-dihyro-8-iodo- 1-methyl-2,5-dioxo- lH-1,4-benzodiazepine]-4-propanoate(2 mmol), 2-(methylaminomethyl)ben~ill,idazole dihydrochloride (3 mmol), DIEA (1.8 mL, 10 mmol), and (Ph3P)2PdC12 (140 mg, 0.2 mrnol) in N-methyl-2-pyrrolidinone (20 mL) is heated to 110~C under CO
balloon for 3 h. The mixture is then concentrated and the residue is purified bysilica gel flash chromatography to give the title compound.
g) 3-[3,4-Dihydro-8-[[[(ber-7imi~1~7Ol-2-yl)methyl]methylamino]ca.bollyl]-l-methyl-2,5-dioxo- 1 H- 1,4-benzodiazepine]-4-propanoic acid A mixture of benzyl-3-[3,4-dihyro-8-[[[(bel,,.i...ifl~7Ql-2-yl)methyl]methylamino]carbonyl3 - 1 -methyl-2,5-dioxo- 1 H- 1,4-benzodiazepine3-4-propanoate (2 mmol) and 10% Pd/C (0.02 g) in ethanol (100 mL) is hydrogenated inan atmosphere of H2 (50 psi3 for 6 h. The catalyst is removed by filtration, and the filtrate is concentrated under vacuo to afford the title compound.
Fx~m~le 62 ~re~aldlion of 3-{4H-imi-l~7Q~1~2-al~1.41benzQdiazepine-5(6H)-l-methyl-6-oxQ-9-rrr(ber~7imi~ 701-2-yl)methyllmethylaminolcarbonyl~ }-4-propanoic acid a) Ethyl N-(2-amino-4-iodobenzoyl)-,B-alanine A m~gneti~lly stirred solution of 4-iodoisatoic anhydride (0.0173 mol)"l3 alanine ethyl ester hydrochloride (0.0173 mol), and dimethylaminopyridine (0.5 g, - 35 0.0041 mol) in pyridine (35 mL) is heated for 2 h at 80~C. The reaction mixture is allowed to cool to RT and concentrated in vacuo. The resulting residue is dissolved in ethyl acetate (100 mL), and washed successively with 10% cupric sulfate (2 x 50 W O 97/24119 PCT~US96/2~748 mL), saturated sodium bicarbonate (1 x 50 rr~ ) and brine (1 x 50 rnL).
Drying (Na2SO4), filtration, concentration, and silica gel chromatography (1:1 EtOAc/hexanes) gives the title compound.
b) Ethyl-3-[3 ,4-dihyro-8-iodo-2,5-dioxo- 1 H- 1 ,4-benzodiazepine]-4-propanoateTo a cold (-30~C) m:~gnPti~z~lly stirred solution of ethyl N-(2-amino-4-iodobenzoyl)-,B-alanine (0.69 mmol), and triethylamine (0.144 g, 1.04 mmol) in methylene chloride (3 rnL) is added slowly a solution of a~-bromoacetyl bromide (0.09 mL, 1.04 mmol) in methylene chloride (2 mL) under argon. The reaction mixture is allowed to warm to RT and stir for 2 h. The mixture is diluted with methylene chloride (40 mL) and wash snccç~ively with 10% citric acid (1 x 50 mT.) and saturated sodium bicarbonate (1 x 50 mL), dried (Na2SO;), filtered, and concentrated in vacuo. The rçsllltin~ residue is dissolved in DMF (3 mL) and added via an addition funnel to a slurry of sodium hydride (25 mg, 1.04 mmol) in DMF (2 mL) which is cooled to 0~C. After 2 h of stirring, the rnixture is poured over an ice cooled solution of 10% citric acid (50 mL) and extracted with ethyl acetate (3 x 40 rnL). The combined extracts are washed with saturated sodium bicarbonate ( 1 x 50 rnL), dried (Na2SO4), filtered, concentrated, and chromatographed on silica gel to afford the title compound.
c) Ethyl-3-[3 ,4-dihyro-8-iodo-2-thione-5-oxo- 1 H- 1 ,4-benzodiazepine~-4-propanoate To a solution of ethyl-3-[3,4-dihyro-8-iodo-2,5-dioxo- 1 H- 1,4-benzodiazepine]-4-propanoate ( 1.0 g, 2.49 mmol) in THF ( 10 mL) at RT and underan atmosphere of nitrogen is added Lawesson's reagent ( 1.0 g), and the reaction is heated at 50~C for 2 h. The reaction mixture is allowed to cool to RT and is concentrated in vacuo. ~ilica gel chromatography (gradient 40-60%
EtOAc/hexanes) gives the title compound.
d) Ethyl-3-[4H-imi-1~7o~1,2-a][1,4]benzodiazepine-5(6H)-1-methyl-6-oxo-9-iodo]-4-propanoate To a vigorously stirred biphasic solution of ethyl-3-[3,4-dihyro-8-iodo-2-thione-S-oxo-lH-1,4-benzodiazepine]-4-propanoate (0.95 g, 2.27 mmol), methyl iodide (0.2 g) and a catalytic amount of tetrabutylammonium hydrogen sulfate in CH2CI2 (10 rnL) and water (10 mL) is added 2 N NaOH (1.2 mL) at RT. After 2 h, the layers are separated and the aqueous layer is washed with CH2Cl2 (2 x 25 rnL).
The combined organic extracts are dried (Na2SO4), filtered and concentrated in vacuo. The resulting residue is dissolved in toluene (10 rnL) and treated with W O 97/24119 PCTr~S96/20748 pl~pa.~yl amine (0.64 mL, 4-fold excess) and pyridine hydrogen chloride (0.23 g, 1 mol eq). The reaction is heated to reflux for 6 h, then was allowed to cool to RT. Concentration and silica gel chromatography (EtOAc) gives the title compound.
s e) Ethyl-3-[4H-imi~l~7~[1,2-a][1,4]benzodiazepine-5(6H)-l-methyl-6-oxo-9-[[[(benzimidazol-2-yl)methyl]methylamino]carbonyl] }-4-propanoate A rnixture of ethyl-3-[4H-imi~1~7O[ l ,2-a] ~ 1,4]benzodiazepine-5(6H)- 1 -methyl-6-oxo-9-iodo]-4-propanoate(2 mmol), 2-(methylaminomethyl)ben7imi(1z~7c-1e(3 mmol), DIEA (1.8 mL, 10 mmol), and (Ph3P)2PdCl2 (140 mg, 0.2 mmol) in N-methyl-2-pyrrolidinone (20 mL) is heated to 110~C under a CO balloon for 3 h. The mixture is then concentrated and the residue is purified by silica gel flash chromatography to give the title compound.
f) 3-[4H-Imidazo[1,2-a][1,4]benzodiazepine-5(6H)-l-methyl-6-oxo-9-[[[(ben7imi~i~7Ql-2-yl)methyl]methylamino]carbonyl]]4-propanoic acid A solution of ethyl-3-[4H-imi~l~7o[l~2-a][l~4]benzodiazepine-5(6H) methyl-6-oxo-9-[[t(ben7imi~1~7ol-2-yl)methyl]methylamino]carbonyl] }-4-propanoate (54 mmol), LiOH H20 (0.79 mmol), THF (5 mL), and water (2 mL) is stirred at RT overnight. The mixture is concentrated, the residue is dissolved in water, and the resulting solution is neutralized with 3 N HCl. The precipitate is collected and dried in vacuo to afford the title compound.
E~xample 63 P~ lion of 4-r4-r2-(lH-ben7~imi~l~7~ol-2-yl)ethyll-l-piperazin piperi~1inP~çetiç acid a) Ethyl 4-[4-[(tert-butoxycarbonyl)]-1-pipe~ lyl]-1-piperi~1inP~et~t~
The title compound is prepared from tert-butyl l-piperazinecarboxylate (Aldrich) and ethyl 4-oxo-1-piperitlinP~ret:ltP (Porter et al. EPA 0 542 363 A2) by NaCNBH3 reductive amination according to the method of Porter et al., EPA 0 542 363 A2.
b) Ethyl 4-(1-~ipe~dzinyl)- 1-piperi(1in~P;Icet~tP
W O 97/24119 PCTrUS96/20748 A solution of ethyl 4-[4-t(tert-butoxycarbonyl)]-1-pi~e~ yl]-l-piperitlin~a~et~t~ and 4 M HCl/dioxane in CH2Cl2 is stirred at RT for 18 h. The reaction mixture is concentrated to give the title compound as the hydrochloride salt.
5 c) 2-[2-Chloroethyl)]be.n7.imirl~7.ole A solution of 2-ben7.imici:~7.Qleethanol and thionyl chloride in CH2Cl~ is heated at reflux for 2 h. The mixture is evaporated to give the title compound.
d) Ethyl of 4-[4-[2-(lH-bPn7imi<1~7.Ql-2-yl)ethyl]-l-pipeld~ yl]-l-10 piperitlinf.~e~slt~
A solution of ethyl 4-(1-pi~ela~ yl)-1-pipericlinP~ret~te, 2-[2-chloroethyl)]ben7imi/i~7.Qle, and Dl~EA in DMF is stirred at RT for 18 h. The mixture is concentrated and purified by chromatography to give the title compound.
e) 4-[4-[2-(lH-Ben7imi(1~7- 1-2-yl)ethyl]-1-~ .dzinyl]-1-piperi~1inP.~reti~ acidA solution of ethyl of 4-[4-[2-(lH-ken7.imiti~7.--1-2-yl)ethyl]-l-pipera_inyl]-l-piperi~linea~et~te. and 1.0 N NaOH in MeOH is stirred at RT. After 18 h, the mixture is neutralized with AcOH, cle.s~ltP.cl through an XAD-2 column, and lyophiii7P.-l to give the title compound.
F,x~mrle. 64 Prep~ration of 1 -hydroxy-4-r4-r3-( 1 H-be~7imi~1~7.ol-2-yl)propyll- 1 -piperazinyll-eyclohex:~neacetic acid a) tert-Butyl l-hydroxy4-[4-[2-(lH-bçn7.imi-1~7.ol-2-yl)propyl~ ;ldzinyl]-cyclohexaneacetate A solution of tert-butyl l-hydroxy-4-(1-pipel~inyl)-cyclohexs~nP~-~et~tP
(EPA O 537 980 Al~, 2-(3-bromopropyl)ben7imi(1~7.ole (J. Org. Chem. 1962, 27, 3U 2165), and DIEA in DMF is stirred at RT for 18 h. The l~ Lul~iS concentrated and purified by chromatography to give the title compound.
b) 1 -Hydroxy-4-[4-[2-~ lH-ben7.imi~l~701-2-yl)propyl]- l-pipera_inyl]-cyclohex:~n~ etic acid A solution of tert-butyl l-hydroxy~-[4-[2-(lH-ben7imi~1~7.ol-2-yl~propyl]-l-yip~,.dzinyl]-cyclohex~nP.~cet~te and 4 M HCl/dioxane in CH~Cl2 is stirred at RT.
After 18 h, the mixture is evaporated to give the title compound.
CA 0224l633 l998-06-26 W O97/24119 PCTrUS96/20748 Ex~mple 6S
Preparation of N-r3-~l-rr2-(2-Ben7imi(1~7olvl)ethyllcarbonyllpiperidinyllcarbonyll-5 ,13-alanine Following the procedures of Beavers et. al., WO 95/25091, Example 1, except ~ubstitl-tin~ (2-ben7imi~701yl)propionic acid for N~'-Boc-D-lys(Cbz)-OH, gives the title compound.
Example 66 Preparation of 2-r~Ben7imi(1~7ol-2-yl)methyll- 5-r2-~carboxy-ethyl)amino~call,ollyll -2.3-dihydro-3-oxo- 1 H-isoindole Following the procedures of rl~Jaldlion 1-12 in Hartman, et al., EP 0 540 334 Al, for the prepa-dlion of l-H-isoindole-5-carboxamide, 2,3-dihydro-N-(2-carboxy-ethyl)-2-[2-(piperidinyl)ethyl]-3-oxo, except substituting 2-(aminomethyl)be.l~ lidazole (Aldrich) for Boc-4-piperidine-2-ethylamine, the title compound is pl~pal~ed.
Bxample 67 Ple~ lion of r3(R)-r2-(be~ 7ol-2-yl)ethyll-2-oxopiperidinyllacetyl-3(R)-methyl -~-alanine a) Methyl 4-(ben7imifl~7Ol-2-yl) butanoate Following the procedure of Example 36(a), except sub~LiLuLillg 1,2-diaminobenzene for the 2,3-diaminopyridine, the title compound is prepared.
b) 4-(Ben7imi~ 7OI-2-yl) butanoic acid Following the procedure of Example 36(b), methyl 4-(benzimidazol-2-yl) butanoate is saponified to afford the title compoud.
c) [3(R)-r2-~Ben7.imi~ ol-2-yl)ethyl]-2-oxopiperidinyl]acetyl-3(R)-methyl-,13-- 35 alanine .
W O 97/24119 PCT~US96/20748 Following the procedure of Duggan et al (J. Med. Chem. 1995, 38, 3332), except using 4-(ben7imid~7Ol-2-yl) butanoic acid instead of (N-Boc-piperidin-4-yl3butanoic acid, the title compound is prepared.
E~ rr~le68 .
Preparation of 4-rrrr2-(benzimidazolyl)methyllcall,unyllmethylaminol-acetyllphenoxyacetic acida) 4-[2-(Boc-methylamino)acetyl]phenol A solution of di-tert-butyl dicarbonate (5.96 g, 27.3 mmol) in 1,4-dioxane (25 mL) was added dropwise at 0~C to a mixture of 4-t2-(methylamino)acetyl]phenol hydrochloride (5.0 g, 24.8 mmol), 1,4-dioxane (30 mL), H2O (25 rnL) and 1.0 N NaOH (25 mL, 25 mmol). After 24 h, the reaction was warmed to RT and stirred for 1.5 h. More 1.0 N NaOH (25 mL, 25 mmol) was added, and the reaction was stirred for an additional 0.5 h at RT, and concentrated.
The residue was diluted with EtOAc (80 mL), and the ll~ix.Lule was acidified to pH 2 using 1.0 M NaHSO4. The res-llsin~ mixture was extracted with EtOAc, and the combined organic layers were washed with H20 and dried (Na2S04). Filtration and concentration gave the title compound (6.49 g, 99%): lH NMR (250 MHz, CDCl3) ~ 6.70-8.05 (m, 4 H), 4.53 (s, 2H), 2.98 (s, 3H), 1.50 (s, 9H).
b) Benzyl 4-[2-(Boc-methylamino)acetyl]phenoxyacetate A mixture of the compound of Example 68(a) (5.04 g, 19.0 mmol) and K2CO3 (2.63 g, 19.0 mmol) in acetone (100 mL) was stirred at reflux under argon for lh. The mixture was cooled to RT and benzyl bromo~etS-~ (5.23 g, 22.8 mmol) was added. The reaction was heated at reflux for 18 h, then was cooled and filtered.
The filter cake was washed with acetone, and the filtrate was concentrated. The residue was dissolved in CH2Cl2 (300 mL) and washed sequentially with H2O (50 mL) and brine (50 mL). Drying ~Na2SO4), concentration, and flash chromatography ( silica gel, 1 :3 EtOAc/hexanes) yielded the title compound (7.28 g, 93%): 1H NMR (250 MHz, CDCl3) o 6.85-7.95 (m, 9 H), 5.23 (s, 2H), 4.71 (s, 2H), - 4.55 (d, 2H), 2.95 (d, 3H), 1.45 (d, 9H).
c) Benzyl 4-r2-(methylamino)acetyl]phenoxyacetate hydrochloride - A mixture of the compound of Example 68(b) (7.26 g, 17.57 mmol) and 4 M
HCl in 1,4-dioxane (150 mL) was stirred for 1 h at RT. Concentration and trituration with Et2O afforded the title compound as a white powder (5.93 g, 97%):
CA 0224l633 l998-06-26 W O 97/24119 PCTrUS96/20748 1H NMR (250 MHz, CD30D) ~ 7.05-8.00 (m, 9 H), 5.23 (s, 2H), 4.88 (s, 2H), 4.65 (s, 2H), 2.80 (s, 3H).
d) Benzyl 4-~[1:[2-S (bçn7imicl~701yl)methyl3carbonyl]methylalI~ino]acetyl~phenoxyacetate A mi~lule of the compound of Example 68(c)(1 mmol), 2-(ben7imicl~7Olyl)acetic acid (1 mmol), EDC (1.5 mmol), and D~ 3 mrnol) in DMF (25 mL) is stirred at RT. The mixture is poured in to 5% NaHCO3 and extracted with EtOAc. The organic phase is washed with H2O, dried (MgSO4) and concentrated. The residue is chromatographed (silica gel) to give the title compound.
e) 4-[[[[2-(Ben7imit1~7Olyl)methyl]carbonyl]methylamino]acetyl]phenoxyacetate The compound of Example 68(d)(1 mmol) and lN NaOH (1.5 mL) in CH30H (20 mL) is stirred and concentrated. The residue is dissolved in H2O, extracted with CH2C12, and the aqueous phase is adjusted to pH 5 with dilute HCl to give the title compound.
Fx~mI?le 69 Pl~;yaldlion of 4-rrrr2-(Benzimidazolyl)methyllcarbonyllmethylaminolacetyll-1.2-l?henylenedioxvdiacetiç acid a) 4-[2-(Boc-methylamino)acetyl~- 1,2-dihydroxybenzene Following the procedure of Example 68(a), except substituting adrenalone hydrochloride (5.0 g, 23.0 mmol) for 4-[2-(methylamino)acetyl]phenol hydrochloride, the title compound (1.2 g, 19%) was prepared following flash chromatography (silica gel, 1: I EtOAc/hexanes): MS (ES) m/e 282.2 [M+H]+.
b) Dimethyl 4-[2-(Boc-methylamino)acetyl]-1,2-phenylenedioxy~ ret~te Following the procedure of Example 68(b), except substituting the compound of Example 69(a) (0.9 g, 3.2 mmol) for the compound of Example 68(a) and methyl bromoacetate (1.23 g, 8.0 mmol) for ~enzyl bromo~cet~e, the title co.llpoul~d (1.11 g, 81%) was prepared: MS (ES) m/e 426.2 [M+H]+.
c) Dimethyl 4-~2-(methylamino)acetyl]- 1,2-phenylenedioxydiacetate hydrochloride 195 CA 0224l633 l998-06-26 W O 97/24119 PCTrUS96~0748 Following the procedure of Example 68(c), except substituting the compound of Example 69(b) (1.1 1 g, 2.6 mmol) for the compound of Example 68(b), the title colllpoulld was l~r~al~;d (1.1 g, qu~..l ;l~tive): MS (ES) m/e 326.0 [M+H]+.
d) Dimethyl 4-[[[[2-(ben7imi-1A7QIyl)methyl]carbonyl]methylarnino]acetyl]-1,2-phenylenedioxyrliAcetAtP
Following the procedure of procedure of Exarnple 68(d), except sub~ ; ugthe ~;ul~ o~-nd of Example 69(c) for the compound of Example 68(c), gives the title 10 compound.
e) 4-[[[[2-(Benzimidazolyl)methyl]carbonyl]methylamino]acetyl]- 1,2-phenylenedioxydiacetic acid Following the procedure of procedure of l~xarnple 68(e), except substituting 15 the compound of Example 69(d) for the compound of Example68(d), gives the title compound.
Fx~mple 70 Preparatisn of N-r3-rrrt2-Ben7imi-1A7olyl)metllyllçarbonyllamirlolbenzoyll-¦3-alar~ine a) Benzyl N-[3-[[t(2-bçn7imi(1:~7olyl)methyl]carbonyl]arl~ino]benzoyl]-,B-AI~ninAte A ~ Y.lul~ of benzyl N-(3-aminobenzoyl)-,B-~IAninAt~ (Alig, et. al., EPA
372486)(1 mmol), (2-b~7imi~l~7Q}yl)acetic acid (1 mmol), EDC (1.5 mmol), and DIEA (3 rnmol) in DMF (25 mL) is stirred at RT. The mixture is poured into 5%
NaHCO3 and extracted with EtOAc. The combined organic phase is washed with H2O, dried (MgSO4) and concentrated. The residue is chromatographed (silica gel)to give the title compound.
b) N-[3-[[[(2-Bçn7imi~1A7olyl)methyl]carbonyl]amino]benzoylJ-~3-alanine A mixture of the compound of Example 70(a)( 1 mmol) and lN NaOH ( 1.5 mL) in CH30H (20 mL) is stirred and coneell~ldled. The residue is dissolved in H2O, extracted with CH2Cl2, and the aqueous phase is adjusted to pH 5 with dilute HCl to give the title compound.
F.~ 7 1 CA 0224l633 l998-06-26 W O 97/24119 PCTrJS96/20748 Pl~al~tion ofrrl-rN-rr(2-Ben7.imi~7.olyl)methyl]carbonyl~tyrosyll-4-piperidinylloxylacetic acid =
S a) tert-Butyl [[l-[N-[[(2-ben7imi~lA7olyl)methyl]c~ln,llyl]tyrosyl]-4 piperidinyl]oxy]acetate A mixture of tert-butyl [( l-tyrosyl-4-piperidinyl)oxy]acetate (Alig, et. al., EPA 372486)(1 mmol), (2-ben7imi~1A7olyl)acetic acid (1 mmol), EDC (1.5 mmol), and DIEA (3 mmol) in DMP (25 mL) is stirred at RT. The mixture is poured into 10 5% NaHCO3 and extracted with EtOAc. The combined organic phase is washed with H2O, dried (MgSO4) and concentrated. The residue is chromatographed (silicagel) to give the title compound.
b) [[l-[N-[[(2-Benzimida_olyl)methyl]call~ollyl]tyrosyl]~-piperidinyl]oxy]acetic15 acid A mixture of the compound of Example 71(a)(1 mmol) and CF3CO2H in CH2CI2 is stirred and concentrated to give the title compound.
F.x~ ?le 72 P-~pal~lion of fS)-4-rrr(2-Benzimidazolyl)methyllcarbonyllglycyll-3-m~thoxycarbonylmethyl-2-oxopipe-d~ine-l-acetic acid Following the procedure of Sugihara, et. al., EP 0529858, Example 59, except substit~-ting (2-ben7imitlA701yl)acetic -acid for 4-amidinobenzoic acid 25 hydrochloride, gives the title compound.
PxArnple 73 Preparation of (3S.5S)-5-rr4-[(2-Ben71midA7olyl)methyllphenylloxymethyl~-3-30 carboxymethyl-2-pyrrolidinone a) 4-[(2-Ben7imi-1A7l-1yl)methyl]phenol Pollowing the general procedure of Wahlgren and Addison, J. Heterocycl.
- Chem., 1989, 26, 541-543, except substitl-ting 4-(hydroxy)phenylacetic acid for 2-35 (hydroxy)phenylacetic acid, gives the title compound.
..
-CA 02241633 1998-06-W O97/24119 PCT~US96/20748 b) (3S,5S)-5-t[4-[~2-Benzimidazolyl)methyl]phenyl3O~y~ hyl]-3-t(tert-butoxycarbonyl)methyl] -2-pyrrolidinone Following the procedure of Hirnmelsbach, et.al., Australian Patent Application AU-A-86926/91, Example 51, substituting the compound of Example 5 73(a) for 4'-cyano-3'-fluoro-4-hydroxy)biphenyl, gives the title compound.
c) (3S,SS)-5-[~4-[(2-Ben7imi-1~7olyl)methyl~phenyl]oxymethyl]-3-carboxymethyl-2-pyrrolidinone The compound of Example 73(b) is treated with CF3CO2H in CH2CI2 to give the title compound.
F~ le 74 Prçparatioll of 1-r~2-Ben7imi~ m~tllyll-3-r4-(2-carboxyethyl)phenyl~-1 5 m~,thoxy-3-pyrrolin-2-one Following the procedures of Linz, et. al., EP 0567968, except sub~ (2-ben7imi-1~701yl)methsln:~min/ for 4-cyano~nilin~, gives the title colllpo~l,ld.
Fx~ ple 75 Plti~,a,~tion of 2-r6-(ben7imi~1~7ol-2-yl) metllylaminocarbonyl)-12.3 tetrahydroisoquinolinyllacetic acid a) 6-Methoxy- 1,2,3 ,4-tetrahydroisoquinoline 6-Methoxy-1,2,3,~tetrahydroisoquinoline is prepared according to the method of D. J. Sall and G. L. Grunewald (J. Med. C~hem. 1987, 30, 2208-2216).
b) Ethyl 2-[6-Methoxy-1,2,3,~tetrahydroisoquinolinyl]acetate A solution of the compound of Example 75(a) ( 1.1 mmol), ethyl chloroacetate (1.17 mmol), and pot~ssillm carbonate (1.17 mrnol) in acetonitrile (10 mL) is stirred for 18 hr. The mixture is then partitioned in a mixture of EtOAc and H20. The organic phase is rotary evaporated to an oil, which is purified by silica gel chromatography to afford the title co~ oulld.
W O 97/2411g PCTrUS96/20748 c) Ethyl 2-[6-Hydroxy-1,2,3,4-tetrahydroisoquinolinyl]acetate A solution of the compound of Example 75(b3 (0.249 g, 1.0 mmol) and boron tribromide ( lM in CH2Cl2, 1.0 mL, 1.0mmol, mL) is stirred at -70 ~C for 2 hr and then stirred at RT for 12 hr. The solution is rotary evaporated to an oil. The - S residue is taken up in EtOAc. EtOAc is washed with water (lX), 5% NaHCO3(2X), water (lX). EtOAc is dried over Mg2SO4, filtered and rotary evaporated to afford the title compound.
d) Ethyl 2-[6-trifluoromethylsulfonyloxy-1,2,3,4-tetrahydroisoquinolinyl]acetateA solution of the compound of Example 75(c) (0.235 g, 1.0 mmol), trifluorosulfonic acid anyhdride (0.23 mL, l.lmmol,) and Et3N (0.32 mL, 1.5 mmol) in CH2Cl2 (S ml)is stirred for 8 hr The solution is rotary evaporated to an oil. The residue is taken up in EtOAc. EtOAc is washed with 5% NaHCO3 (2X), water (lX).
EtOAc is dried over Na2SO4, filtered and rotary evaporated to afford the title lS compound.
e) Ethyl 2-[6-carboxy-1,2,3,4-tetrahydroisoquinolinyl~acetate A solution of the compound of Example 75(d) (0.367 g, 1.0mmol), pall~ m(II)bis-acetate (0.022 g, 0.1mmol,), triphenylphosphine (0.262 g, 1.0mmol), diisopropylamine (0.34 mL, 2.5 mmol), NMP (S mL), in aqueous ~mmonillm carbonate (10%) is stirred for 8 hr under an atmosphere of carbon monoxide. The solution is rotary evaporated to an oil. The residue is purified by silica gel chromatography to afford the title compound.
f) Ethyl -[6-(ben7imicl~7Ol-2-yl)methylaminocarbonyl)-1,2,3,4-tetrahydroisoquinolinyl]acetic acid A solution of the compound of Example 75(e) (0.263 g, 1.0 mmol), the compound of (2-ben~ 7Olyl)acetic acid (0.34g, 1.0 mmol), EDC (0.19lg, 1.0 - mmol), HOBt (0.151 g, 1.0 mmol) and triethylamine (0.235 mL, 2.0 mmol) in DMF(7 mL) is stirred for 8 hr. The solution is rotary evaporated to an oil. The }esidue is purified by silica gel column to afford the title compound.
CA 0224l633 l998-06-26 W O97/24119 PCTrUS96/20748 g) 2-[6-(ben7imi~1~7ol-2-yl)methylarninocarbonyl)-1,2,3,4-tetrahydroisoquinolinyl]acetic acid A solution of the compound of Example 75(f) ( 0.42 g, 1.0 mmol) in aqueous 1 N sodium hydroxide (1.5 mL, 1.5 mmol) and ethanol ( 5 mL) is stirred for 8 hr.The solution is rotary evaL,ol~led to an oil. The residue is purified by silica gel column to afford the title compound.
Example 76 Pre~?~ration of 2-r6-fbell~i...id~7ol-2-yl)methylaminoca~l,ollyl)-l-oxo-1.2.3~4-tetrahydroisoquinolir~ acetic acid a) 6-Methoxy- 1 -oxo- 1,2,3,4-tetrahydroisoquinoline 6-Methoxy-l-oxo-1,2,3,4-tetrahydroisoquinoline is p~ d according to the method of D. J. Sall and G. L. Grunewald, J. Med. Chem. (1987), 30, 2208-2216.
b) Ethyl 2-[6-Methoxy-l-oxo-1,2,3,4-tetrahydroisoquinolinyl]acetate A mixture of the compound of Example 76(a) (0.39 mmol) and NaH (0.17 g, 0.43 mmol, 60% oil dispersion) in THF (5 rnL) is heated to reflux for 1 hr and then allowed to cool to room ~e~ el~ture. Ethyl chloroacetate (0.43 mmol) is added tothe mixture, and the mixture is allowed to stir for 1 hr. The mixture is quenched with water (10 rnL) and washed with EtOAc (2X 15 rnL). The organic layers are combined, washed with water (10 rnL) and rotary evaporated to an oil. The residue is purified by silica gel chromatography to afford the title compound.
c) Ethyl 2-[6-Hydroxy-l-oxo-1,2,3,4-tetrahydroisoquinolinyl~acetate A solution of the compound of Example 76(b) (0.263 g, 1.0 mmol) and boron tribromide (lM solution in CH~Cll, 1.1 mL) is stirred at -70 ~C for 2 hr and then at RT for 4 hr. The solution is rotary evaporated to an oil. The residueis taken up in EtOAc. EtOAc is washed with water (lX), 5% NaHCO3 (2X), water (lX).
W O97/24119 PCTrUS96/20748 EtOAc is dried over MgSO4, filtered and rotary evaporated to afford the title compound.
~. d) Ethyl 2-[6-trifluoromethylsulfonyloxy-1-oxo-1,2,3,4-- 5 tetrahydroisoquinolinyl]acetate A solution of the compound of F.~mple 76(c) (3.4 mmol) and trifluorosulfonic acid anyhdride (3.4 mmol, mL) in pyridine (5 mL) is chilled at 0~
and allowed to warm to room temperature for 1 hr. The mixture is quenched with water (5 mL~ and washed with EtOAc (2X 7 mL). The organic layers are combined, washed with water (7 rnL) and rotary evaporated to an oil. The residue is purified by silica gel chromatography to afford the title compound.
e) Ethyl 2-[6-carboxy-1-oxo-1,2,3,4-tetrahydroisoquinolinyl~acetate A solution of the compound of Example 76(d) (0.23 g, 1.0 mmol), palladium(II)bis-acetate (0.026 g, 0.1 mmol), triphenylphosphine (0.262 g, 1.0 mmol), diisopropyla~nine (0.23 mL, 2.0 mmol), NMP (7 mL), in aqueous ammonium carbonate "(10 %) is stirred for 8 hr under an atmosphere of carbon monoxide. The solution is rotary evaporated to an oil. The residue is purified by silica gel chromatography to afford the title compound.
f) Ethyl -[6-(bc~ 7Ol-2-yl)methylaminocarbonyl)- 1,2,3,4-tetrahydroisoquinolinyl]acetic acid A solution of the compound of Example 76(e) (0.34 g, 1.0 mmol), the compound of (2-ben~ idazolyl)acetic acid (0.43 g, 1.0 mmol), EDC (0.191 g, 1.0 mmol), HOBt (0.15 g, 1.0 mmol) and triethylamine (0.234 mL, 2.3 mmol) in DMF(8 mL) is stirred for 8 hr. The solution is concentratred. The residue is purified by silica gel chromatography to afford the title compound.
~lt~ tively, a solution of the compound of Example 76(d) (0.23 g, 1.0 mmol), p~ m(II)bis-acetate (0.026 g, 0.1 mmol), triphenylphosphine (0.262 g, - 1.0 mmol), diisopropylarnine (0.25 mL, 2.1 mmol), NMP (7 mL), and the compound W O 97/24119 PCTrUS96/20748 of Interm~ t~ A (0.31 g, 1.0 mmol) in aqueous ammonium carbonate ~10%) is stirred for 8 hr under an atmosphere of carbon monoxide. The solution is rotary evaporated to an oil. The residue is purified by silica gel chromatography to afford the title compound.
g) 2-[6-((ben7imi~l~7ol-2-yl)methylaminocarbonyl)-1,2,3,4-tetrahydroisoquinolinyl]acetic acid A The solution of the compound of Example 76(f) (0.25 g, 1.0 mmol) in aqueous 1 N sodium hydroxide (1.5 mL, 1.5 mmol) and ethanol (8 mL) is stirred for 10 8 hr. Solution is rotary evaporated to an oil. The residue is purified by silica gel to afford the title compound.
Rx~mple 77 Preparation of 2-r6-((be~ 7~1-2-yl)methylcarbonyl:~mino)tetralinlacetic acid a ) tert-butyl-5-amino-tetraline-2-acetate Tert-butyl-5-amino-tetraline-2-acetate is prepared according to the methods described in M. J. Fisher, et al. (Scheme 12 and Example 28, parts A-D, EO
0635492, Jan. 25, 1995).
b) 2-[6-((benzimidazol-2-yl)methylcarbonylamino)tetralin]acetate A solution of the compound of Example 77 (a) (0.261 g, 1.0 mmol), 2-(aminomethyl)ben7imid~7O1e(0.256g, l.Ommol),EDC(0.19lg, l.Ommol) 1-hydroxybenzotriazole hydrate (0.152 g, 1.0 mmol), and triethylamine (0.234 mL, 2.1 mmol) in DMF (5 mL) is allowed to stir for ~ hr. The solution is rotary evaporated to an oil. The residue is purified by silica gel chromatography to afford the title compound.
A solution of the crude ester amide (0.32 g, 1.0 mmol) and trifluoroacetic acid (5 mL) in methylene chloride (~ mL) is allowed to stir for 1 hr. The solution is CA 0224l633 l998-06-26 W O97/24119 PCT~US96/20748 rotary evaporated to an oil. The residue is treated with Et2O. Filtration and drying in vacuo afforded the title compound.
;, F.x~mple 78 ~l~;palalion of 2-r6-((benzimidazol-2-yl)methylaminocarbonyl)tetr~linJacetic acid a) Ethyl-5-hydroxy-tetraline-2-acetate The compound Ethyl-5-hydroxy-tetraline-2-acetic acid is prepared according to the method of M. J. Fisher et al. (EP 0635492, Scheme 6 and Example 20, parts A-D, p. 71).
b) Ethyl-5-trifluoromethylsulfonyloxy-tetraline-2-acetate A solution of the compound of Example 78(b) (0.321g, 1.0 mmol) in CH2Cl~
(10 mL) is cooled to O~ C.Trifluoromethylsulfonic acid anhydride ( 0.125 mL, 1.1mmol) is added. The solution is stirred for 2 hr. The solution is rotary evaporated to an oil. The residue is taken up in EtOAc. EtOAc is washed with water (lX), 5%
NaHCO3, water (lX). EtOAc is dried over MgSO4, filtered. Filtrate is rotary evaporated to afford the title compound.
c) Ethyl 6-carboxy-tetraline-2-acetate.
A solution of the compound of Example 78(c) (0.26 g, 1.0 mmol), palladium(II)bis-acetate (0.023g, 0.1 mmol), triphenylphosphine ~0.262g, 1.0 mmol), diisopropylamine (0.245 mL, 2.1 mmol), NMP (10 mL ), in aqueous 25 ammonium carbonate (10%) is stirred for 8 hr under an ~tmosphere of carbon monoxide. The solution is rotary evaporated to an oil. The residue is purified by silica gel chromatography to afford the title compound.
- d) Ethyl -[6-((ben7imi~l~7ol-2-yl)methylaminocarbonyl)-tetraline-l-acetate A solution of the compound of Example 78(c) (0.34 g, 1.0 mmol), the compound of (2-ben7imicl~olyl)acetic acid (0.32g, 1.0 mmol), EDC (0.191 g, 1.0 W O 97/24119 PCTrUS96120748 mmol), HOBt ~0.152 g, 1.0 mmol) and triethylamine (0.23 mL, 2.1 mrnol3 in DMF(6 mL) is stirred for 8 hr. The solution is rotary evaporated to an oil. The residue is purified by silica gel chromatography to afford the title compound.
~It.orll~tively, a solution of the compound of Example 78(b) ~0.34 g, 1.0 mmol), palladium(II)bis-acetate (0.023g, 0.1 mmol), triphenylphosphine (0.262g, 1.0 mmol), diisopropylarnine (0.23 mL, 2.1 mmol), NMP (10 mL), and the compound of IntermPrli~te A (0.32 g, 1.0 mmol), in aqueous ammonium carbonate (10%) is stirred for 8 hr under an zitm~spht~re of carbon monoxide. The solution is rotary evaporated to an oil. The residue is purified by silica gel chromatography to afford the title compound.
e) 2-~6-((be.~i."~ 7Ol-2-yl)methylaminocarbonyl)-tetraline-2-acetic acid.
A solution of the compound of Example 78(d) (0.31 g, 1.0 mmol) in aqueous 1 N sodium hydroxide (1.5 mL, 1.5 mmol) and ethanol ( 5 mL) is stirred for 8 hr.The solution is rotary evaporated to an oil. The residue is purified by silica gel chromatography to afford the title compound.
Fx~mple 79 Preparation of 2-r5-((be~7,i l ll;dzl~ol-2-yl)methylcarbonylamino)-benzofuranl-.
proplomc acld a) Ethyl 1-carboxymethyloxy~-nitrosalicylaldehyde A solution of 1-hydroxy~-nitrosalicylaldehyde (Aldrich) ( 0.167 g, 1.0 mmol), ethyl bromoacetate (0.166 g, 1.0rnmol) potSlc~inm carbonate ( 0.276 g, 2.0mmol) and sodium iodide (0.015 g, 0.1 mmol) in THF ( 10mL) is heated to 80 ~Cfor 24 hr. The solution is rotary evaporated to an oil and the residue is purified by silica gel chromatography to afford the title compound.
b) Ethyl 2-carboxy-5-nitrobenzofuran W O 97/24119 PCTrUS96/20748 A solution of the compound of Example 79(a) (0.229 g, 1.0 mmol), DBU (0.152 g, 1.0 mmol) in ethyl alcohol ( 10 mL) is allowed to stir at RT for 18 hr.
The solution is rotary evaporated to an oil and the residue is treated with EtOH (10 mL). The solution is bubbled with HCl.gas for 2 minllt~s and refluxed for 5 hr. The - 5 solution is rotary evaporated to an oil. EtOAc is added and washed with water (2X), 5% citric acid (2X), water (1~), 5% NaHCO3.(2X) and water (lX). EtOAc is rotary evaporated to afford the title compound.
c) Ethyl 2-(2-carboxy)ethylene-5-nitrobenzofuran A cold solution (-78 ~C) of the compound of Example 79(b) ( 0.235 g, 1.0 mmol)in THF (5 mL) is treated with DiBAL ( 1.0 M in THF, 1.0 mL, 1.0 mmol).
The solution is stirred at -78 ~C for 30 min~ltes and RT for 3 hr. The solution is treated with CH3COOH (3 mL) followed by water (2 rnL). The solution is rotary evaporated to an oil and treated with toluene to azeotrope off the acetic acid. Drying in vacuo afforded the crude aldehyde.
A solution of the phosphonate ester (0.224 g, 1.0 mmol),in THF (5 mL) is treated with sodium hydride (60% suspension in mineral oil, 0.04 g, 1.0 rnmol)at 0~
C for 1 hr. To the solution is added the aldehyde (0.235 g, 1.0 mmol). The solution is stirred at RT for 18 hr. The solution is rotary evaporated to an oil and the residue is purified by silica gel chromatography to afford the title compound.
d) tert-butyl 2-[5-amino-benzofuranyl]propionate A solution of the compound of Example 79(c) ( 0.261 g, 1.0 rnrnol) in ethanol ( 5 mL) containing 10% palladium-on-carbon(0.026 g, 10% wt) is hydrogenated at 45 psi for 1 hr. The solution is filtered through Celite and the- filtrate is rotary evaporated to an oil. Silica gel cl~ t~ graphy affords the title compound.
e) 2-~5-((bçn7imicl~7Ql-2-yl)methylcarbonylamino)-benzofuran]-propionic acid W O 97/24119 ~CT~US96/20748 A solution of the compound of Example 79(d) ( 0.263 g, 1.0 mmol), EDC ( 0.191 g, 1.0 mmol), 1-hydroxybenzotriazole ( 0.150 g, 1.0 mmol), the compound of 2-(aminomethyl)benzimidazole ( 0.234 g, 1.0 mmol) and triethylamine (0.288 mL, 2.0 mmol) in DMF (5.0 mL) is allowed to stir for 18 hr. The solution is rotary evaporated to an oil and the residue is purified by silica gel chromatography to afford the title compound.
A solution of the crude ester ( 0.263 g, 1.0 mmol) in MeOH (3.0 mL) is treated with 1 N NaOH ( l .S mL, l .S mmol) and water (2 ml). The solution is stirred at RT for 18 hr. The solution is rotary evaporated to an oil and purified by reversed phase chromatography to afford the title compound.
F.x~?le 80 Preparation of 2-rS-((benzirnidazol-2-yl)methylcarbonylamino)-2.3-dihydro-ben7Ofilranl-propionic acid a) Ethyl-2-[S-amino-2,3-dihydro-benzofuranyl3propionate In the chromatographic purification of the compound of Example 79(d) the -20 title compound is also obtained.
b) 2-[5-(6-arninopyridinyl-2-methylcarbonylamino)-benzofuran]-propionic acid A solution of the compound of Example 80(a) (0.263 g, 1.0 mmol), EDC
(0.19lg, 1.0 rnmol), 1-hydroxybenzotriazole (0.lSg, 1.0 m~nol), the compound of 2-(aminomethyl)ben7imi.1~701e (0.32 g, 1.0 mmol) and triethylamine (0.288 mL, 2.0 mmol) in DMF (5.0 mL) is allowed to stir for 18 hr. The solution is rotary evaporated to an oil and the residue is purified by silica gel chromatography toafford the title compound.
A solution of the crude ester ( 0.290g, 1.0 rnmol) in MeOH (5.0 mL) is treated with lN NaOH (1.2 mL, 1.2 mmol) for 18 hr. The solution is rotary .
W O 97/24119 PCT~US9~/20748 evaporated to an oil and purified by reversed-phase chromatography to afford the title compound.
-Fxample 81 S
Preparation of 2-r5-(6-aminopyridinyl-2-nlethylaminocarbonyl)-benzofuranl-propionic acid a) 2-Ethyloxycarbonyl-5-(tert-butyl-dimethylsilyloxy)-benzofuran A solution of 2-Ethyloxycarbonyl-5-(hydroxy)-benzofuran is prepared via the procedure described in M. L. Denny, et al. (EP 0655439, 31,5,95) (0.206 g, 1.0 mmol), tert-butyl-dimethylsilylchloride (0.23 mL, 1.0 mmol) and imidazole ( 0.34 g, 1.0 mmol) in THF is allowed to stir for 4 hr. The solution is rotary evaporated to an oil. EtOAc is added and washed with water. EtOAc is rotary evaporated to afford the title compound.
b) 2-Hydrocarbonyl -5-(tert-butyl-dimethylsilyloxy)-benzofuran A cold solution (-78 ~ C) of the compound of Example 81 (a) ( 0.35 g, 1.0 mmol)in THF (5 mL) is treated with DiBAL ( 1.0 M in THF, 1.0 mL, 1.0 mmol).
The solution is stirred at -78 ~ C for 30 minutes and RT for 3 hr. The solution is treated with CH~COOH (3 mL) followed by water (2 mL). The solution is rotary evaporated to an oil and treated with toluene to azeotrope off the acetic acid. Drying in vacuo afforded the aldehyde.
c) Ethyl 2-[5-(tert-butyl-dimethylsilyloxy)-benzofuran]-acrylate A solution of the phosphonate ester (0.224 g, 1.0 mmol),in THF (5 rnL) is treated with sodium hydride (60% s-lcpen~ion in mineral oil, 0.04 g, 1.0 mmol)at 0"
C for 1 hr. To the solution is added the above aldehyde(0.235 g, 1.0 mmol). The solution is stirred at RT for 18 hr. The solution is rotary evaporated to an oil and the residue is purified by silica gel chromatography to afford the title compound.
W O 97/24119 PCT~US96J20748 d) 2-Ethyl 2-~-5-(hydroxy)-benzofuran]-propionate A mixture of the compound of Example 81(c) (0.234g, 1.2 mmol) and 10%
p~ m-on-carbon (0.023 g, 10% wt) in EtOH( S mL).is hydrogenated at 50 psi 5 for 1 hr. Filtration through Celite and concentration afforded.the ester (0.169 g, 56%).
A solution of the crude ester (0.34 g, 1.0 mmol) and tetraethylammonium fluoride (0.149 g, 1.0 mmol) in THF (10 mL) is allowed to stir at RT for 18 hr. The 10 solution is rotary evaporated to an oil and purified by silica gel ch~ atography.
e) 2-Ethyl 2-[-5-(trifluoromethylsulfonyloxy)-benzofuran]-propionate A solution of the compound of Example 81(d) (0.366 g, 1.0 mmol) and Et3N(0.23 mL, 1.5 mmol) in CH2Cl2 (10 mL) at 0 "C is treated with trifluoromethylsulfonic acid anhydride (0.21 mL, 1.1 mmol). After 2 hr solution is rotary evaporated to an oil. The residue is taken up in EtOAc. EtOAc is washed successively with water(lX), 5% Na~ICO3 (2X), water (lX). EtOAc is dried over MgSO4, and filtered. Filtrate is rotary evaporated to afford the title compound.
20 f) 2-Ethyl 2-[-5-(carboxy)-benzofuran3-propionate A solution of the compound of Example 81(e) (0.366g, 1.0 mmol), p~ m(II) bis-acetate (0.023g, 0.1 mmol), triphenylphospine (0.262g, 1.0 mmol), diisopropylethylamine (0.23 mL, 2.1 mmol), NMP (7 mL) in aqueous sodium bicarbonate (10%, 6 mL) is allowed to stir. The solution is rotary evaporated to an 25 oil. The residue is purified by silica gel chromatography to afford the title compound.
g) 2-Ethyl 2-~-5-(6-(6-aminopyridinyl-2-methylaminocarbonyl)-benzofuran]-propionate W O 97/24119 PCT~US96/20748 A solution of the compound of Example 81(f) (0.366 g, 1.0 mmol), the compound of (2-ben7imid~701yl)acetic acid (0.23g, 1.0 mmol), EDC (0.191 g, 1.0mmol), HOBt (0.152g, 1.0mmol) and triethylamine (0.235 mL, 2.1mmol) in DMF(8 mL) is stirred for 8 hr. The solution is rotary ev~poldled to an oil. The 5 residue is purified by silica gel chromatography to afford the title compound.
Alt~-rn~tively, a solution of the compound of Example 81(e) (0.366g, l .Ommol), palladium(II)bis-acetate (0.023g, 0.1 mmol), triphenylphosphine (0.262g, 1.0 mmol), diisopropylamine (0.23 mL, 2.1 mmol), NMP (10 mL ), and the compound of Intern~ t(~ A (0.32g, 1.0 mmol) in aqueous ammonium carbonate (10%, 10 mL) is stirred for 8 hr under an atmosphere of carbon monoxide. The solution is rotary evaporated to an oil. The residue is purified by silica gel chromatography to afford the title compound.
h) 2-[-5-(6-(6-aminopyridinyl-2-methylaminocarbonyl)-benzofuran]-propionic acid A solution of the col"pou~d of Example 81(g) (0.366g, 1.0 mmol) in aqueous 1 N sodium hydroxide (1.5 mL, 1.5 mmol) and ethanol ( 8 rnL) is stirred for 8 hr. The solution is rotary eva~o,~l~d to an oil. The residue is purified by silica gel chromatography to afford the title compound.
Ex~n~le 8~
Preparation of 2-rS-((benzi",idazol-2-yl)methylaminocarbonyl)-2.3-dihydro-ben7:r~furan~-propionic acid a) 2-tert-Butyl 2-[-5-(hydroxy)-2,3-dihydro-benzofuran]-propionate The chromatographic purification in Example 81 (d) also provides the title compound.
b) 2-tert-Butyl 2-[-5-(trifluoromethylsulfonyloxy)-2,3-dihydro-benzofuran]-propionate W O 97/24119 PCT~US96/207~.
A cold solution of the compound of Example 82(a) (0.28 g, 1.0 mmol) and Et3N (0.23 mL, 2.1 mmol) in CH2Cl2 (5 mT.)is treated with trifluoromethylsulfonic acid anhydride (0.15 mL, 1.1 mmol) for 2 hr. The solution is rotary evaporated to an oil. The residue is taken up in EtOAc. EtOAc is washed 5 with water (lX), 5% NaHCO3(2X), water (lX). EtOAc is dried over MgSO4, filtered, and filtrate is rotary evaporated to afford the title compound.
c) 2-tert-Butyl 2-[-5-(carboxy)-2,3-dihydro-benzofuran]-propionate A solution of the compound of Example 82(b) (0.24 g, 1.0 mmol), 10 p~ um(II) bis-acetate (0.023g, 0.1 mmol), triphenylphospine (0.262g, 1.0 mmol), diisopropylethylamine (0.23 rnL, 2.1 mmol), NMP (8 mL ) in aqueous sodium bicarbonate (10 mL, 10%) is allowed to stir at RT for 8 hr. The solution is rotary evaporated to an oil. The residue is purified by silica gel chromatography to afford the title compound.
d) 2-tert-Butyl 2-[-5-(6-(6-aminopyridinyl-2-methylaminocarbonyl)-2,3-dihydro-benzofuran] -propionate A solution of the compound of Example 82(c) (0.366g, 1.0 mmol), the compound of (2-be~ 7olyl)acetic acid ( 0.23g, 1.0 mmol), EDC (0.19lg, 1.0 20 mmol), HOBt (0.152g, 1.0 ~nol) and triethylamine (0.23 mL, 2.1 mmol) in DMF(8mL) is stirred for 8 hr. The solution is rotary evaporated to an oil. The residue is purified by silica gel chromatography to afford the title compound.
~It~ tively, a solution of the compound of Example 82(b) (0.366g, 1.0 25 mmol), p~ m(II)bis-acetate (0.023 g, 0.1 mmol), triphenylphosphine (0.2~2 g, 1.0 rnmol), diisopropylamine (0.23 mL, 2.1 mmol), NMP (10 mL), and the compound of Interm~ te A (0.23 g, 1.0 mmol), in aqueous ammonium carbonate (10%, 10 mL) is stirred for 8 hr under an atmosphere of carbon monoxide for 8 hr.
The solution is rotary evaporated to an oil. The residue is purified by silica gel 30 chromatography to afford the title compound.
W O97/24119 PCT~US96/20748 e) 2-[-5-(6-((ben7imit1~7Ql-2-yl)methylaminocarbonyl)-2,3-dihydro-benzofuran]-propionic acid . A solution of the compound of Example 82(d) (0.37 g, 1.0 mmol) in aqueous 1 N sodium hydroxide ( 1.5 mL, 1.5 mmol) and ethanol (8 mL) is stirred for 8 hr.- 5 The solution is rotary eva~oldted to an oil. The residue is purified by silica gel chromatography to afford the title compound.
.
Example 83 lQ ~lcpald~ion of 1 -r( lH-ben7imi(1~7ol-2-yl)methyll-3-{4r(2-ethoxycarbonyl)ethyl)lphenyl }-3-oxo-imidazolidine a) 2-Aminomethyl-l-p-toll-enl-sl-lfonyl-benzimidazole A solution of 2-Methylben7imi~701e (1 mmol), p-toluenesulfonyl chloride (1.05 mmol) and Et3N (1.05 mmol) in water (2 mT ) and THF (1 mL) is allowed to stir for 4 hr. The ~ L~nc is concentrated by rotary evaporation and diluted withwater (10 mL). The solution is washed with EtOAc (2X 15 mL). The organic layers are combined, washed with water (S mL) and rotary evaporated to an oil.
A solution of the aforementioned oil (2-Methyl- 1 -p-toluenesulfonyl-benzimidazole) (1 mmol) and N-bromosuccinimide (1.05 mrnol) in CCl4 (10 mL) is allowed to reflux for 18 hr. Upon cooling a white solid precipitates from solution.
The solid is filtered, triturated with CCl4 and dried in vacuo to a solid.
A solution of Boc2NH (1 mmol) and pot~sil-m hydroxide ~1 mmol~ in ethanol (S mL) is allowed to stir for 1 hr. Anhydrous ethyl ether (15 mL) is added and the mixture is filtered to afford the salt as a white solid. A solution of the aforementioned solid 2-bromomethyl-l-p-toluenesulfony!bl~n7i..~ 701e (1 mmol) and (Boc)2N~ Kt (mmol) in THF (10 mT.)is to stir at 60~ for 18 hr. The mixture is 30 rotary evaporated to an oil.
A solution of the 2-[bis-(tert-butyloxycarbonyl)aminomethyl]- l-p-toluenesulfonylben7im~ 701e (1 mmol) in TFA (1.1 mmol) and CH2Cl2 is allowed to stir for 1 hr. The solution is rotary ev~o-dled to an oil, which is purified by 35 chromatography to afford the title compound.
W O97/24119 PCT~US96/20748 b) Ethyl 2-[4-(2-hydroxyethylamino)phenyl]propionate This compound is prepared following the procedure of F. ~imm~l~bach et al.
~Example V, p. 44, EP 0587134, Sept. 8, 1993), in which glycolaldehyde dimer (Aldrich) (mmol) is added to a solution of methyl 2-(4-aminophenyl)propionate 1 (1 mmol) in aqueous acetonitrile (pH 6-7) (mL), followed by sodiumcyanoborohydride (1.1 mmol), and the mixture is allowed to stir for 1 hr. The lluhLIul~ is rotaryevaporated to an oil, and the residue is dissolved in a mixture of ice water and ethyl acetate. The water layer is neutralized with 4 N sodium hydroxide and washed with ethyl acetate. The organic phase is rotary evaporated to an oil. A solution of the oil in ethyl acetate is purified on a silica gel column to give the title compound.
c) N-[(l-p-toluenesulfonyl-lH-be~ 7~l-2-yl)methyl]-N~-hydroxyethyl-N~
{4[(2-ethoxycarbonyl)ethyl)]phenyl } -urea This compound is Lnc~cd following the procedures of F. Himmelsbach et al. (EP 0587134, Sept. 8, 1993 and EP 0612741, Feb.21, 1994), in which a solution of the compound of Example 83(a) (1 mmol) and phosgene (1.1 mmol) in THF (20 mL) is allowed to stir at -20~ for 20 minutes The compound of Example 83(b) (1.0mmol) is added to the solution and the r~cllltin~ mixture is allowed to stir for 18 hr.
The resulting solution is rotary evaporated. A solution of the residue in ethyl acetate is washed with 5% citric acid, followed by water. The organic phase is rotary evaporated to an oil. . A solution of the oil in ethyl acetate is purified on a silica gel column to give the title compound.
d) N'-[(l-p-toluenesulfonyl-lH-ben7imifl~7ol-2-yl)methyl]-N3-{4t(2-ethoxycarbonyl)ethyl)]phenyl } -2-oxo-imidazolidine This compound is prepared following the procedures of F. Himmelsbach et al. (Example III, EP 0587134, Sept. 8, 1993 and EP 0612741, Feb.21, 1994), in which a solution of the compound of Example 83(c) (1 mmol), mf-th~n~sulfonylchloride (1.2 mmol) and triethylamine (1.2 mrnol) in methylene chloride (5 ml) is allowed to stir at 0~ for 1 hour. The ~ lul~; is partitioned in a mixture of water and methylene chloride. The organic phases are combined and dried over anhydrous sodium sulfate and rotary evaporated.
A solution of the residue and sodium iodide (1.1 mmol) in acetone (5 mL) is heated to reflux for 3 hr and then rotary evaporated to an oil. Potassium-bis(trimethylsilyl)azide (1.2 mmol) is added to a solution of the residue in DMF (5 mL), cooled to 0~. The solution is allowed to warrn to room t~ln~el~lul~ over 30 212 , PCT~US96120748 min. and the rotary evaporated to an oil. The residue is partitioned in a ul~ of water and methylene chloride. The organic phases are combined and dried over anhydrous sodium sulfate and rotary evaporated. A solution of the oil in ethyl acetate is purified on a silica gel column to give the title compound.
e) N~ H-bçn7im~ 7ol-2-yl)methyl]-N3-{4[(2-carboxyl)ethyl)]phenyl}-2 imicl~olidine A solution of the compound of Example 83(d) ( 1 mmol) in THF (S mL) and 1 N sodium hydroxide (1.2 mL, 1.2 mmol) is allowed to stir for 18 hr. The mixture 10 is neutralized with conc. hydrochloric acid and purified on a silica gel column to give the title compound.
F,~ m,ple 84 Parenteral Dosage Unit Composition A preparation which contains 20 mg of the compound of Example 1 as a sterile dry powder is prepared as follows: 20 mg of the compound is dissolved in 15 mL of distilled water. The solution is filtered under sterile conditions into a 25 ml multi-dose ampoule and lyophilized. The powder is reconctit-lte~l by addition of 20 mT of 5% dextrose in water (DSW) for intravenous or intr~mllccul~r injection. The dosage is thereby det~,rmin~l by the injection volume. Subsequent dilution may be made by addition of a metered volume of this dosage unit to another volume of DSW for injection, or a metered dose may be added to another m~c,h~nicm for en~ g the drug, as in a bottle or bag for IV drip infusion or other injection-infusion system.
Exarnple 85 C)ral Dosa~e Unit Composition A capsule for oral ~f1mini,ctration is prepared by mixing and rnilling 50 mg of the compound of Example 1 with 75 mg of lactose and 5 mg of m~gnl~,citlm stearate.
The rçs--ltin~ powder is screened and filled into a hard gelatin capsule.
F,x:~mple 86 Oral r~osa~e Unit Com,~osition A tablet for oral a~1minictration is prepared by mixing and gr~n~ ting 20 mg of sucrose, 150 mg of calcium sulfate dihydrate and 50 mg of the compound of J
CA 0224l633 l998-06-26 Example 1 with a 10% gelatin solution. The wet granules are screened, dried, mixed with 10 mg starch, 5 mg talc and 3 mg stearic acid; and co~ r~ssed into a tablet.
The above description fully discloses how to make and use the present - invention. However, the present invention is not limited to the particular embodiments described hereinabove, but includes all modifications thereof withinthe scope of the following claims. The various references to journals, patents and other publications which are cited herein comprises the state of the art and areincorporated herein by reference as though fully set forth.
r
N ~CO2CH3 NH2 H ~ H 2 - ,¦, b Q~ N ~ CO2CH3 ~c,d CO2CH3 Q--~ ~~/
'¦, e CO2H
~ ~~
a) HO2CCH2Ph-4-CH2CH~CO2CH3, Ph2POCl, Et3N, DMAP, THF; b) NaH, DMF, BrCH2CO2CH3; c) KOt-Bu, THF, DMF; d) KOt-Bu, CH3I. DMF; e) LiOH, THF, H20.
The procedures of Linz, et. al., EP 0567968, are used to prepare XIX-5, 10 except (2-ben~.imi~1~7.olyl)m~th~n~mine is su~ led for 4-cyanoaniline.
_g~
CA 0224l633 l998-06-26 W O 97/24119 PCT~US96/20748 Sch~me XX
MeO ~C~ MeO ~C~ b NH N ~ CO2Et HO ~ N ~ co2Et R-NHCO ~ e HO2C ~
N ~ CO2Et N ~ CO2Et R-NHCO~C~ R = [~C \~
N ~ CO2Et S a) ClCH2CO2Et, Et3N, DMF; b) BBr3; c) (CF3SO)20; d) CO, Pd(OAc)~, PPh3, DIEA, NMP, NH4HCO3, H20; e) H2N-R, EDC / HOBt, DIEA, DMF; f) H2N-R, CO, Pd(OAc)2, PPh3, DIEA, NMP, NH4HCO3, H20; g) lN NaOH, HOEt Scheme XX provides a method for the preparation of 1,2,3,4-tetrahydroisoquinoline compounds as exemplary fibrinogen receptor antagonists, as described in M. J. Fisher et al. (EP 0635492, Jan. 25, 1995). Accordingly, a 6-methoxy-3,4-dihydroisoquinoline, such as compound XX-~I is prepared by the method described by D. J. Sall and G. L. Grunewald (J. Med. Chem. 1987, 30, - 8~ -.
CA 0224l633 l998-06-26 W O 97/24119 PCT~US96/20748 2208-2216). The isoquinoline is treated with a haloacetic acid ester in the presence of a tertiary amine to afford the 2-acetic acid ester, as exemplified by compound XX-2. The 6-methoxy compound is converted into the corresponding 6-hydroxy compound by methods known in the art, for example with BBr3, which is converted 5 into the triflate with trifluorosulfonic acid anhydride. p~ m catalyzed carbonylation affords the 6-carboxy compound, such as compound XX-5, which is then condensed with an amine, as exemplified by (2-ben7.imi~1~7.olyl)acetic acid, employing a standard amide bond forming reagent to give the desired amide, such as compound XX-6. Saponification affords XX-7. Alternatively, the palladium 10 catalyzed carbonylation reaction with the triflate, exemplified by compound XX-4, may be trapped with said aminomethyl compound to provide, after saponification, the corresponding 6-(2-bçn7.imi~ 7.Qlyl)methylaminocarbonyl compound, XX-7.
Sch~otne XXI
HO ~ CF3SO3~
~N~CO2Et ~N~CO2Et 3 O ~ 4 ~
R-NHCO~ e HO2C~
~N~CO2Et ~N ~CO2Et 6 ~ 5 ~
lg R-NHCO~,~ R = ~N~
N ~ CO2Ft ~J--NH
7 o a) 1. LiN(TMS)2, 2. ClCH2CO2Et, DMF; b) BBr3; c) (CF3SO2)20; d) CO, Pd(OAc)2, PPh3, DIEA, NMP, NH4HC03, H20; e) H2N-R, EDC / HOBt, DIEA, DMF; f) H2N-R
CO, Pd(OAc)2, PPh2, DIEA, NMP, NH4HCO3, H20; g) lN NaOH, HOEt -8~-W O 97/24119 PCT~US96/20748 Scheme XXI provides a method for the pl~al~lion of 3,4-dihydroisoquinolin- l-one compounds as exemplary fibrinogen receptor antagonists, as described M. J. Fisher et al. (~P 0635492, Jan. 25, 1995). Accordingly, the 1-oxo compound XXI-1, prepared by the method described by D. J. Sall and G. L.
Grunewald (J. Med. Chem. 1g87, 30, 2208-2216), is treated with a base, such as LiN(TMS)~, and a haloacetic acid eser to give a 2-acetic acid ester, as exemplified by compound XXI-2. The 1-oxo compound is then employed in the analogous series of reactions deployed in Scheme XX, sub~ g the corresponding 1-oxo analog, as shown in Scheme XXI, to provide XXI-7. As in Scheme XX, alternatively, the p~ m catalyzed carbonylation reaction with the triflate, exemplified by compound XXI-4, may be trapped with an amine to provide, after saponification, the amide, XXI-7.
Sch~mf XXII
H2N~C~ a, b R-CONH~
N ~ CO2tBu N ~ CO2H
~N
~ ~ NH
a) RCO-X; b) TFA / CH2C12 Scheme XXII provides a method for the pr~alaLion of 6-acylaminotetraline compounds as exemplary fibrinogen receptor antagonists, as described M. J. Fisher et al. (;EP 0635492, Jan. 25, 1995). Accordingly, a 6-arnino-2-tert-butyloxyca.1.onyl-tetral-1-one, exemplified by compound XXII-l, which is prepared according to the methods described in M. J. Fisher et al. (EP 0635492, Jan. 25, 1995), is con-l~.n.~ed with an activated derivative of a carboxylic acid, such as the CA 0224l633 l998-06-26 W O 97/24119 PCTrUS96/20748 activated derivative of (2-bPn7imi-~7olyl3acetic acid, to provide, after deesterification, the amide, XXII-2.
Sch~rnP XXlII
HO ~[~G CF3SO3 ~0~ CO2Et R-NHCO~ c HO2C~
COzEt ~ CO2Et 1 e R-NHCO = N~
a) CF3SO2O; b) CO, Pd(OAc)2, PPh3, DIEA, NMP, NH4HCO3, H2O; c) H2N-R, EDC
/ HOBt, DIEA, DMF; d) H2N-R, CO, Pd(OAc)2, PPh3, DIEA, NMP, NH4HCO3, 10 H2O; e) lN NaOH, HOEt Scheme XXIII provides a method for the plc;paldlion of 6-aminoacyltetraline compounds as exemplary fibrinogen receptor antagonists, as described M. J. Fisher et al. (EP 0635492, Jan. 25, 1995). Accordingly, an ethylo~ycall,onylmethyl-6-15 hydroxy-tetral-l-one, exemplified by compound XXIII-l, which is prepared according to the methods described in M. J. Fisher et al. (EP 0635492, Jan. 25, W O 97/24119 PCTrUS96120748 1995), is treated with triffic anhydride to provide the triflate, as exem~ ed bycompound XXIII -2, which is employed in a p~ m catalyzed carbonylation reaction to afford a carboxylic acid, such as compound XXIII -3, which is then condensed with an amine to provide, after deesterifi-~tion, the 6-aminoacyl S compound, XXIII -5. ~AltPrn~tively~ the palladium catalyzed carbonylation reaction with the triflate exemplified by compound XXIII -2, may be trapped with said amine compound to provide, after saponification, the corresponding 6-aminoacyl compound, XXII~ -5.
Scheme XXIV
02N~ EtO IP~o S CO2Et 4 ,/
e H2N ~ + ~ CO2Et 1 f~9 1 f'9 R-CONH~ RCONH~ ~ CO2H
.~ N \
a) BrCH2CO2Et, KzCO3~ NaI; b) 1. DBU, EtOH, 2. HCl, EtOH; c) DiBAL, -78 ~ C; d) 15 NaH, THF; e)Hz, 10% Pd-C; f) R2CO-X; g)lN NaOH, MeOH
-8~-W O97/24119 PCTrUS96120748 Scheme XXIV provides a method for the preparation of 5-acylaminobenzofuran and S-acylaminodihydrobenzofuran compounds as exemplary r fibrinogen receptor antagonists, as described in M. L. Denney, et al. (~P 0655439, 31/5/95). Accordingly, a 4-nitrosalicylaldehyde, exemplified by compound XXIV-1 5 , is treated with a haloacetic acid ester to give the phenoxyacetic acid ester, exemplified by compound XXIV -2. A 2-alkoxycarbonylfuran, exemplified by compound XXIV -3, is obtained by treating the aldehyde with base, for example with DBU. The 2-alkoxycarbonyl group is reduced to the aldehyde, for example with DiBAB. Wittig reaction affords the 2-acrylate ester, exemplified by compound 10 XXIV -4, which is reduced to the benzofuran-2-propionic acid ester, exemplified by compound XXIV -5 and the dihydrobenzofuran-2-propionic acid ester, exemplified by co~ )o-llld XXIV -6. The amine XXIV -~ is then condensed with an activated derivative of a carboxylic acid to provide, after deesterification, the amide 5, XXIV -8. ~lt~rnzltively, the amine XXIV -6 is condensed with an activated derivative of a 15 carboxylic acid to provide, after dççstçrifiç~tion, the amide, XXIV -7.
-8~-W O97/24119 PCTrUS96/20748 Schem~ XXVa HO ~ CO2Et ~ CO2Et b,c a-l a-2 TBDMSO ~ CHO TBDMSO ~
EtO2P ~ a-~ ~ CO2Et ~e,f HO ~ CO2Et ~ CO2Et a-6 a-7 a)l.TBDMS-Cl, imid~ole; b)DiBAl-H, -78 ~C, d) NaH, THF; e) H2, 5% Pd-C; f) Et4N+ F -CA 0224l633 l998-06-26 W O 97/24119 PCTrUS96/20748 Scheme XXVb HO ~ CF3SO3 ~ CO2Et a-6 b-1 d/
~' I b R-NHCO~ CO~EtCO2Et b-3 b-2 l e R-NHCO~co2H R- [~NH
b-4 a) ~CF3SO2)20; b) CO,Pd(OAc)2, PPh3, DTF.A, NMP, NH4HCO3, H20; c) H2N-R, EDC / HOBt, DIEA, DMF; d) H2N-R, CO, Pd(OAc)2, PPh2, DIEA, NMP, NH4HCO3, H20 e) lN NaOH, EtOH
W O 97/24119 PCT~US96/20748 SchemP XXVc CO2Et a-7 c-1 ~ 1 R-NHCO ~ CO2Et CO2Et l e R-NHCO~co2H R =[~NH
a) (CF3SO2)20; b) CO, Pd(OAc)2, PPh3, DlF.A, NMP, NH4HC03, H20; c) H~N-R, EDC / HOBt, DIEA, DMF; d) H2N-R, CO, Pd(OAc)2, PPh2, DIEA, NMP, NH4HCO3, H20 e) lN NaOH, BtOH
Scheme XXV provides a method for the ~rt;p~dtion of 5-aminoacylbenzofuran and 5-aminoacyldihydrobenzofuran compounds as exemplary fibrinogen receptor antagonists, as described in M. L. Denney, et al. (EP 0655439, 31/5/95). Accordingly, a 5-hyd~ ybellzofuran-2-carboxylic acid ester, such as compound XXVn-l, prepared in the manner of M. L. Denney, et al. (~P 0655439, 31/5/95), is treated with TBDMS-Cl to provide the TBDMS delivalive of the ester,XXVa-2. The ester is reduced to an aldehyde, such as co,llpolJ.ld XXVa-3. Wittig 9~
W O 97/24119 PCT~US96/2074.~.
reaction affords the acrylic acid ester, XXVa-4. Catalytic reduction affords thebenzofuran-2-acetic acid ester and the dihydl. bell of uran-2-acetic acid ester.r Cleavage of the silyl ether group of each ester, by methods known to the art, affords either the benzofuran-2-acetic acid ester, XXVa-~, or the dihydrobenzofuran-2-S acetic acid ester, XXVa-6.
~ As seen in Scheme XXVb and XXVc, each alcohol in turn may be converted to a carboxylic acid via p~ m catalyzed carbonylation, such as compound XXVb-2 or XXVc-2, which is then condensed with an amine to provide, after ~lees~.; rication, the amide XXVb-4 or XXVc-4. Alternatively, the p~ lm 10 catalyzed carbonylation reaction with the triflate exemplified by compound XXVb-1, or XXVc-1, may be trapped with said aminomethyl compound to provide, after deesterification, the corresponding 6-aminoacyl compound, XXVb-4 or XXVc-4.
Amide coupling reagents as used herein denote reagents which may be used 15 to form peptide bonds. Typical coupling methods employ carbo-liimicles, activated anhydrides and esters and acyl halides. Reagents such as EDC, DCC, DPPA, PPA, BOP reagent, HOBt, N-hydroxysuccinimi(le and oxalyl chloride are typical.
Coupling methods to form peptide bonds are generally well known to the art.
The methods of peptide synthesis generally set forth by Bodansky et al., THE
20 PRACTICE OF ~ SYNTHESIS, Springer-Verlag, Berlin, 1984, Ali et al. in J. Med. Chem., 29, 984 (1986) and J. Med. Chem., 30, 2291 (1987) are generally illustrative of the technique and are incorporated herein by reference.
Typically, the amine or aniline is coupled via its free arnino group to an a~L.,~,iate carboxylic acid substrate using a suitable carbodiimide coupling agent, 25 such as N,N' dicyclohexyl carbodiimide (DCC), optionally in the presence of catalysts such as l-hydroxybenzotriazole (HOBt) and dimethylamino pyridine (DMAP). Other methods, such as the formation of activated esters, anhydrides or acid h~licles, of the free carboxyl of a suitably protected acid substrate, and subsequent reaction with the free amine of a suitably protected amine, optionally in 30 the presence of a base, are also suitable. For example, a protected Boc-amino acid or _9~
W O 97/24119 PCTrUS96/20748 Cbz-amidino benzoic acid is treated in an anhydrous solvent, such as methylene chloride or tetrahydrofuran(THF), in the presence of a base, such as N-methyl morpholine, DMAP or a trialkylamine, with isobutyl chlc,loroll,late to form the "activated anhydride", which is subsequently reacted with the free amine of a second 5 protected amino acid or aniline.
The compounds of formula (XIX) and (XX) are commercially available or are prepared by methods known in the art such as illustrated herein disclosed insta~ldard reference books, like the COMPENDIUM OF ORGANIC S~rHETIC METHODS, Vol. I-VI (Wiley-Interscience). A generally applicable route to ben7imifl~7oles is disclosed in Nestor et al, J. Med. Chem. 1984, 27, 320. Representative methods for preparing compounds of formula (XX) are also common to the art and may be found, for instance, in EP-A 0 381 033.
Acid addition salts of the compounds are prepared in a standard manner in a suitable solvent from the parent compound and an excess of an acid, such as 15 hydrochloric, hydl~lvll~ic, hydrofluoric, sulfuric, phosphoric, acetic, trifluoroacetic, maleic, succinic or methanesulfonic. Certain of the compounds form inner salts or zwitterions which may be acceptable. Cationic salts are prepared by treating theparent compound with an excess of an ~lk~lint? reagent, such as a hydroxide, carbonate or alkoxide, containing the ~lupliate cation; or with an appl.,p~iate 20 organic amine. Cations such as Li+, Na+, K+, Ca++, Mg++ and NH4+ are specific examples of cations present in ph~rms~reutir~lly acceptable salts.
- This invention also provides a pharm~-~entical composition which comprises a compound according to formula (I)-(V) and (XXI)-(XXII) and a ph~rm~reutically acceptable carrier. Accordingly, the compounds of formula (I)-(V) and (XXI)-25 (XXII) may be used in the m~nuf~tllre of a m~-lic~m~nt. Pharmaceutical compositions of the compounds of formula (I)-(V) and (XXI)-(XXII) prepared as hereinbefore described may be form~ te~l as solutions or lyophilized powders forparenteral ~rlmini~tration~ Powders may be recon.ctitnte~1 by addition of a suitable diluent or other ph~rm~entic~lly acceptable carrier prior to use. The liquid 30 fi~rm~ tion may be a buffered, isotonic, aqueous solution. Examples of suitable diluents are normal isotonic saline solution, standard 5% dextrose in water or _ g W O 97/24119 PCTrUS96t20748 buffered sodium or ammonium acetate solution. Such formulation is especially suitable for parenteral ~rlmini~tration, but may also be used for oral ~rlmini~tration or cont~inP~l in a metered dose inhaler or nebulizer for in~llffl~tion It may be desirable to add excipients such as polyvinylpyrrolidone, gelatin, hydroxy cellulose, acacia, polyethylene glycol, m~nnitol, sodium chloride or sodium citrate.
Alternately, these compounds may be encapsulated, tableted or prepared in a emulsion or syrup for oral ~lmini~tration Ph~rm~eeutiç~lly acceptable solid or liquid carriers may be added to enh~nre or stabilize the composition, or to facilitate plG~ald~ion of the composition. Solid carriers include starch, lactose, calcium sulfate dihydrate, terra alba, m~gn~ m stearate or stearic acid, talc, pectin, acacia, agar or gelatin. Liquid carriers include syrup, peanut oil, olive oil, saline and water.
The carrier may also include a sllct~inP~l release m~teri:~l such as glyceryl monostearate or glyceryl distearate, alone or with a wax. The amount of solid carrier varies but, preferably, will be between about 20 mg to about 1 g per dosage unit. The pharmaceutical pl~l,aldLions are made following the conventional techniques of pharmacy involving milling, mixin~, gr:~n~ ting, and compressing, when necessary, for tablet forms; or milling, mixing and filling for hard gelatin capsule forms. When a liquid carrier is used, the ~lepaldlion will be in the form of a syrup, elixir, emulsion or an aqueous or non-aqueous suspension. Such a liquid fnrmnl~tion may be ~flmini.~tered directly p.o. or filled into a soft gelatin capsule.
For rectal ~lmini~tration, the compounds of this invention may also be combined with excipients such as cocoa butter, glycerin, gelatin or polyethyleneglycols and molded into a suppository.
The compounds described herein are antagonists of the vitronectin receptor, and are useful for treating diseases whe~ the underlying pathology is attributable to ligand or cell which interacts with the viLIoile~;lin receptor. For instance, these compounds are useful for the treatment of diseases wherein loss of the bone matrix creates pathology. Thus, the instant compounds are useful for the tre~tment of ostoeporosis, hyperparathyroidism, Paget's disease, hypercalcemia of mz~ n~3ncy,- 30 osteolytic lesions produced by bone m~t~t~ , bone loss due to immobilization or sex hormone deficiency. The compounds of this invention are also believed to have 9~
W O 97/24119 PCTrUS96/20748 utility as ~ntitl~mor, anti-angiogenic, ~ntiinfl~mm~tory and anti-mPt~t~tic agents, and be useful in the treatment of atherosclerosis and restenosis.
The compound is ~lmini~tered either orally or L)a-ellLe,dlly to the patient, in a manner such that the concentration of drug is sufficient to inhibit bone resorption, or S other such indication. The ph:lrm~re~lti(~l composition cont~ining the peptide is ~lmini~tered at an oral dose of between about 0.1 to about 50 mg/kg in a manner conci~tent with the condition of the patient. Preferably the oral dose would be about 0.5 to about 20 mg/kg. For acute therapy, parenteral ~r~ . alion is prefeilcd. An intravenous infusion of the peptide in 5% dextrose in water or normal saline, or a 10 similar formulation with suitable excipients, is most effective, although an cc~ r bolus injection is also useful. Typically, the parenteral dose will be about 0.01 to about 100 mg/kg; preferably between 0.1 and 20 mg/l~g. The compounds are ~lmini~tered one to four times daily at a level to achieve a total daily dose of about 0.4 to about 400 mg/kg/day. The precise level and method by which 15 the compounds are ~lmini.~tf red is readily fletermin~d by one routinely skilled in the art by comparing the blood level of the agent to the concentration required to have a therapeutic effect.
The compounds may be tested in one of several biological assays to determine the concentration of compound which is required to have a given 20 I)h~ ological effect.
Tnhi'~it JI~ of vilrv~ binding Solid-Phase [3H7-sK&F-107260 Binding to av~3: Human placenta or human platelet av~B3 (0.1-0.3 mg/mL) in buffer T (cont~ining 2 mM CaC12 and I %
octylglucoside) was diluted with buffer T collt;1i.. i.. g 1 mM CaC12, 1 mM MnC12, 1 mM MgC12 (buffer A) and 0.05% NaN3, and then imm~ t~ly added to 96-well ELISA plates (Corning, New York, NY) at 0.1 rnL per weIl. 0.1 - 0.2 ~g of ocv,133 was added per well. The plates were incubated overnight at 4~C. At the time of the experiment, the wells were washed once with buffer A and were incubated with 0.1 mL of 3.5% bovine serum alburnin in the same buffer for 1 hr at room W O 97/24119 PCT~US96/20748 telllpeldLuie. Following in~nb:~tion the wells were aspirated completely and washed twice with 0.2 mL buffer A.
Compounds were dissolved in 100% DMSO to give a 2 mM stock solution, which was diluted with binding buffer (15 mM Tris-HCl (pH 7.4), 100 mM NaCl, 1 mM CaC12, 1 mM MnC12, 1 mM MgC12) to a final compound concentration of 100,uM. This solution is then diluted to the required final compound concentration.
Various concentrations of unlabeled antagonists (0.001 - 100 ~M) were added to the wells in triplicates, followed by the addition of 5.0 nM of [3H]-SK&F-107260 (65 -86 Ci/mmol).
The plates were incubated for 1 hr at room lelllpeldLul~. Following incubation the wells were aspirated completely and washed once with 0.2 mL of ice cold buffer A in a well-to-well fashion. The receptors were solubilized with 0.1 mL
of 1% SDS and the bound [3H]-SK&F-107260 was determin~-1 by liquid scin~ tion counting with the addition of 3 mL Ready Safe in a Beckman LS Liquid Scintill~tic~n Counter, with 40% efficiency. Nonspecific binding of [3H3-SK~F-107260 was ~et~rmined in the presence of 2 ,uM SK&F-107260 and was consistently less than 1 % of total radioligand input. The ICso (concentration of the antagonist to inhibit 50% binding of [3H]-SK&F-107260) was determined by a nonlinear, least squares curve-fitting routine, which was modified from the LUNDON-2 program.
The Ki (dissociation constant of the antagonist) was calculated according to theequation: Ki = ICso/( l + L/Kd), where L and Kd were the concentration and the dissociation constant of [3H]-SK&F-107260, respectively.
Compounds of the present invention inhibit vitronectin binding to SK&F
107260 in the concentration range of about 0.001 to 50 micromolar.
Compounds of this invention are also tested for in vitro and in vivo bone resorption in assays standard in the art for ev~ ting inhibition of bone formation, such as the pit formation assay disclosed in EP 528 587, which may also be performed using human osteoclasts in place of rat osteoclasts, and the ovarectomi7ed - rat model. described by Wronski et al., Cells and Materials 1991, Sup. 1, 69-74.
_95 W O 97124119 PCT~US96/20748 Vascular smooth linuscle cell ~ ..lion assay Rat or human aortic smooth muscle cells were used. The cell migration was monitored in a Transwell cell culture chamber by using a polycarbonate membrane with pores of 8 um (Costar). The lower surface of the filter was coated with vitronectin. Cells were suspended in DMEM supplemented with 0.2% bovine serum albumin at a concentration of 2.5 - 5.0 x 106 cells/mL, and were pretreated with test compound at various concentrations for 20 min at 20~C. The solvent alone was used as control. 0.2 mL of the cell suspension was placed in the upper c~ nlent of the chamber. The lower co~ lllent contained 0.6 rnL of DMEM supplemented with 0.2% bovine serum albumin. lncubation was carried out at 37~C in an atmosphere of 95% air/5% CO2 for 24 hr. After incubation, the non-migrated cellson the upper surface of the f1lter were removed by gentle scraping. The filter was then fixed in methanol and stained with 10% Giemsa stain. Migration was measuredeither by a) counting the number of cells that had rnigrated to the lower surface of the filter or by b) extracting the stained cells with 10% acetic acid followed by ~terrninin~ the absorbance at 600 nM.
PARATHYROIDE~TOMIZED RAT MODEL
Each experimental group consists of 5-6 male Sprague-Dawley rats. The rats are parathyroidectomized (by the vendor, Taconic Farms) 7 days prior to use. Twenty four hours prior to use, circ~ tin~ ionized calcium levels are measured in whole blood immediately after it has been withdrawn by tail venipuncture into hep~rini7f r1 tubes. Rats are included if ionized Ca level (measured with a Ciba-Corning model 634 calcium pH
analyzer) is _1.2 mM/L. The rats are then put on a diet of calcium-free chow anddeionized water. At the start of the experiment the rats weigh approximately 100g.
Baseline Ca levels are measured and the rats are ~irnini~tered control vehicle (saline) or compound (dissolved in saline) as a single intravenous (tail vein) bolus injection followed r im mf.~ t~ly by a single ~ubc~lt:~nPous injection of either human paldlhyl~id hormone 1-34 peptide (hPI'Hl-34, dose 0.2mg/kg in saline/0.1% bovine serum albumen, Bachem, Ca) _9~_ W O97/24119 PCT~US~6120748 or the PTH vehicle. The calcemic response to PTH (and any effect of compound on this response) is measured 2h after co~ oulld/PTH allmini~tration.
.
RAT ULNA DRII~T MODEL
Each experimental group consists of 8-10 male Sprague-Dawley or Wistar rats of aL~plo~illlately 30-40g body weight at the start of the e~elilllent. The agent being tested is ~rlmini~trred by an a~prop.iate route as single or multiple daily doses for a period of seven days. Prior to ~lmini~tration of the first dose, the rats are given a single dose of a 10 fluorescent marker (tetracycline 25mg/lcg, or calcein lOmg/lcg) that labels the position of bone forming surfaces at that point in time. After dosing of compound has been completed, the rats are killed and both forelimbs are removed at the elbow, the foot is removed at the ankle and the skin removed. The sample is frozen and mounted vertically on a microtome chuck. Cross sections of the mi(l~h~ft region of the ulna are cut in the 15 cryostat. The rate of bone resorption is measured morphom~-tric~lly in the medial-dorsal portion of the cortical bone. The mea~ule.llellt is done as follows: the amount of bone resorbed at the periosteal surface is equal to the t1ict~nre by which the periosteal surface has advanced towards the fluorescent label which had been incorporated at the endosteal bone formation surface on day zero; this fli~t~nre is calculated by subtracting the width of 20 bone between the label and the periosteal surface on day 7 from the width on day zero; the resorption rate in microns per day is calculated by dividing the result by 7.
HUMAN OSTEOCLAST RESORPTION ASSAY ("PIT ASSAY") 25 ~ Aliquots of osteoclastoma-derived cell suspensions are removed from liquid nitrogen strorage, warmed rapidly at 37~C and washed x l in RPMI- 1640 m~ m by centrifugation (lOOOrpm, 5 mins at 4~C).
~ Aspirate the medium and replace it with murine anti-HLA-DR antibody, diluted 1:3 in RPMI-1640 ml rlillm Incubate for 30 mins on ice and mix the cell suspension .~
frequently.
_ W O 97/24119 PCTrUS96/20748 ~ The cells are washed x2 with cold RPMI-1640 by c~ L irugation (1000 rpm, 5 rnins at 4~C) and the cells are transferred to a sterile 15 rnl centrifuge tube. The number of mononuclear cells are enumerated in an il~l~roved Nellh~ r counting chamber.
S
~ Sufficient m~n~tic beads (5 / mononuclear cell), coated with goat anti-mouse IgG, are removed from their stock bottle and placed into 5 ml of fresh mP~ lm (this washes away the toxic azide preservative). The m~ lm is removed by immobilizing the beads on a magnet and is replaced with fresh medium.
~ The beads are mixed with the cells and the suspension is incub~t~cl for 30 mins on ice. The suspension is mixed frequently.
~ The bead-coated cells are immobilized on a magnet and the rem~ining cells 15 (osteoclast-rich fraction) are dec~nt~d into a sterile 50 ml centrifuge tube.
~ Fresh mto-lillm is added to the bead-coated cells to dislodge any trapped osteoclasts.
This wash process is repeated xlO. The bead-coated cells are discarded.
20 ~ The osteoclast~s are enullleldt~d in a counting chamber, using a large-bore disposable plastic pasteur to charge the chamber with the sample.
~ The cells are pelleted by centrifugation and the density of osteoclasts ~dj-lste~l to 1.5xlO4/ml in EMEM m~ lm, supplemented with 10% fetal calf serum and 25 1.7gllitre of sodium bicarbonate.
~ 3ml aliquots of the cell suspension ( per tre~tmP~t) are ~ieC~ntl ~i into 15mlcenkifuge tubes. The cells are pelleted by centrifugation.
30 ~ To each tube 3m1 of the a~p~ .;ate treatment are added (diluted to 50 uM in the EMEM medium). Also included are ~pl~,pliate vehicle controls, a posiLive conkol gg W O 97/24119 ~T~US96/20748 (87MEM1 diluted to 100 ug/ml) and an isotype control (IgG2a diluted to 100 ug/ml). Incubate at 37~C for 30 mins.
~, ~ 0.5ml aliquots of the cells are seeded onto sterile dentine slices in a 48-well plate - 5 and incubated at 37~C for 2 hours. Each treatment is screened in quadruplicate.
., ~ The slices are washed in six changes of warm PBS (lO ml / well in a 6-well plate) and then placed into fresh tre:~tmf~nt or control. Incubate at 37~C for 48 hours.
10 Tartrate l~- ~;C~nt acid Ph~3s~ ce (TRAP) Pl~ce-lul~ (selective stain for cells of the osteoclast lineage).
~ The slices are washed in phosphate buffered saline and fixed in 2% gluteraldehyde (in 0.2M sodium cacodylate) for 5 mins.
~ They are washed in water and incubated in TRAP buffer for 5 mins at 37~C.
~ Following a wash in cold water they are incubated in cold acetate buffer / fast red garnet for 5 mins at 4~C.
~ Excess buffer is aspirated, and the slices are air dried following a wash in water.
~ The TRAP positive osteoclasts are enumerated by bright-field microscopy and are then removed from the surface of the dentine by sonication.
~ Pit volumes are dele~ ed using the Nikon/Lasertec ILM2 1 W confocal microscope.
Human o~leo~lr--l le3~ ~io.. and adhesion assavs 30 Pit resorption and adhesion assays have been developed and standardized usingnormal human osteoclasts derived from osteoclastoma tissue. The osteoclast _99_ W O 97/24119 PCT~US96/20748 population is negatively selected from the osteoclastoma cell suspensions using magnetic beads (Dynal Inc, NY). These beads are coated with a murine monoclonal antibody that recognizes a hurnan class II major histocompatihility antigen that is present on a large number of mononuclear cells in the cell suspensions. The cells 5 that express this antigen, and consequently bind the beads, are removed from the mixture of cells using a m~n~t The osteoclast-rich suspension is then ready to use in the assays det~ below.
Resor~lio.- sssay (with ELISA readout) 10 Enriched preparations of osteoclasts are preincubated for 30 minllfes at 37~C with test compound (4 doses) or controls. They are then seeded onto bovine cortical bone slices in wells of a 48-well tissue culture plate and are in~ubat-~-l for a further 2 hours at 37nC. The bone slices are washed in six changes of warm phosphate buffered saline (PBS), to remove non-adherent cells, and are then returned to wells of a 48 15 well plate con~ining fresh compound or controls. The tissue culture plate is then incubated for 48 hours at 37"C. The supern~t~n~c from each well are aspirated into individual tubes and are screened in a cornpetitive ELISA that detects a collagen peptide that is released during the resorption process. This is a commercially available ELISA (Osteometer, Denmark) that contains a rabbit antibody that 20 specifically reacts with an 8-amino acid se~uence (Glu-Lys-Ala-His- Asp-Gly-Gly-Arg) that is present in the carboxy-tçrmin~l telopeptide of the al-chain of type I
collagen. The results are expressed as % inhibition of resorption colllpal~;d to a vehicle control.
25 Adhesion assay Osteoclastoma-derived osteoclasts are preincubated with compound (4 doses) or controls at 37~C for 30 minutes. The cells are then seeded onto osteopontin-coated slides (human or rat osteopontin, 2.5ug/ml) and incubated for 2 hours at 37~C. Non adherent cells are removed by washing the slides vigorously in P~S and the cells30 r~m~ining on the slides are fixed in acetone. The osteoclasts are stained for tartrate-resistant acid phosphatase (TRAP), a selective marker for cells of this phenotype, and are enumerated by light microscopy. The results are expressed as % inhibition of adhesion colll~alcd to a vehicle control.
-10~
.
W O 97/24119 PCTrUS96120748 Inhibition of RGD-m~ t.-d GPIIb-IIIa binding pllrific~tion of GPIIb-ma Ten units of ollt~l~tP-l, washed human platelets (obtained from Red Cross) were lyzed by gentle stirring in 3% octylglucoside, 20 mM Tris-HCl, pH 7.4, 140 mM NaCl, 2 mM CaC12 at 4~C for 2 h. The lysate was centrifuged at lOO,OOOg for 1 h. The sUpprn~t~nt obtained was applied to a 5 mL lentil lectin sepharose 4B
column (E.Y. Labs~ preequilibrated with 20 mM Tris-HCl, pH 7.4, 100 mM NaCl, 2 mM CaC12, 1 % octylglucoside (buffer A). After 2 h incubation, the column was washed with 50 rnL cold buffer A. The lectin-retained GPIIb-IIIa was eluted withbuffer A containing 10% dextrose. All procedures were performed at 4~C. The GPIIb-IIIa obtained was >95% pure as shown by SDS polyacrylamide gel electrophoresis .
Incorporation of GPIIb-IIIa in Liposomes.
A mixture of phosphatidylserine (70%) and phosphatidylcholine (30%) (Avanti Polar Lipids) were dried to the walls of a glass tube under a stream of nitrogen. Purified GPIIb-IIIa was diluted to a final concentration of 0.5 mg/rnL and mixed with the phospholipids in a protein:phospholipid ratio of 1:3 (w:w). The mixture was resuspended and sonicated in a bath sonicator for 5 min. The mixturewas then dialyzed overnight using 12,000- 14,000 molecular weight cutoff dialysis tubing against a 1000-fold excess of 50 mM Tris-HCl, pH 7.4, 100 mM NaCl, 2 mM
CaC12 (with 2 changes). The GPIIb-IIIa-cont~ining liposomes wee centrifuged at 12,000g for 1~ min and resuspended in the dialysis buffer at a final protein concentration of approximately 1 mg/mL. The liposomes were stored at -70C until needed.
Competitive Binding to GPIIb-IIIa The binding to the fibrinogen receptor (GPIIb-ma) was assayed by an indirect competitive binding method using [3H]-SK&F-107260 as an RGD-type ligand. The binding assay was performed in a 96-well filtration plate assembly (Millipore Corporation, Bedford, MA) using 0.22 um hydrophilic durapore membranes. The wells were precoated with 0.2 mL of 10 ,ug/mL polylysine (Sigma Chemical Co., St. Louis, MO.) at room temperature for 1 h to block nonspecific 35 binding. Various concentrations of unlabeled ben7~ 7~rines were added to the -1~1-CA 0224l633 l998-06-26 W O 97/24119 PCT~US96/20748 wells in quadruplicate. [3H]-SK&F-IQ7260 was applied to each well at a final concentration of 4.5 nM, followed by the addition of 1,ug of the purified platelet GPIIb-IIIa-cont~ining liposomes. The mi~ulcs were incubated for 1 h at room tel,,pcldture. The GPIIb-ma-bound [3H]-SK&F-107260 was seperated from the S unbound by filtration using a Millipore filtration manifold, followed by washing with ice-cold buffer (2 times, each 0.2 rnL). Bound radioactivity rem:~ining on the filters was counted in 1.5 mL Ready Solve (Beckm~n Instr lm~nt~, Fullerton, CA) in a Beckman Liquid Scintillation Counter (Model LS6800), with 40% efficiency.
Nonspecific binding was ~et~ in.o~t in the presence of 2 ,uM unlabeled SK&F-107260 and was con~i~te-ntly less than 0.14% of the total radioactivity added to the samples. All data points are the mean of quadruplicate ~leterrnin~tions.
Competition binding data were analyzed by a nonlinear least-squares curve fitting procedure. This method provides the IC50 of the antagonists (concentration of the antagonist which inhibits specific binding of [3H]-SK&F-107260 by 50% at equilibrium). The IC50 is related to the equilibrium dissociation constant (Ki) of the antagonist based on the Cheng and Prusoff equation: Ki =
IC50/(l~L/Kd), where L is the concentration of t3H~-SK&F-107260 used in the co~llpeLiLive binding assay (4.5 nM), and Kd is the dissociation constant of r3H]-SK&P-107260 which is 4.5 nM as ~ rmin~l by Scatchard analysis Compounds of the present invention inhibit the vitronectin binding to SK&P
007260 with a Ki at the vitronectin receptor that is about ten-fold greater than that for the fibrinogen receptor. Preferred compounds have a Ki at the vitronectin receptor that is thirty-fold greater than that at the fibrinogen receptor. The most preferred compounds have a Ki at the vitronectin receptor that is a hundred-foldgreater than that at the fibrinogen receptor.
The examples which follow are intended in no way to limit the scope of this invention, but are provided to illustrate how to make and use the compounds of this invention. Many other embodiments will be readily a~cnt to those skilled in the art.
W O 97/2411g PCTrUS96/20748 G~o,n~ral Nuclear magnetîc resonance spectra were recorded at either 250 or 400 MHz using, respectively, a Bruker AM 250 or Bruker AC 400 spectrometer. CDC13 is deuteriochloroform, DMSO-d6 is hPxAfle~ riodimethylsulfoxide, and CD30D is tetradeuteriomethanol. Chemical shifts are reported in parts per million (o) downfield from the internal standard tetramethylsilane. Abbreviations for NMR
data are as follows: s=singlet, d=doublet, t=triplet, q=quartet, m=multiplet, dd=doublet of doublets, dt=doublet of triplets, ~ arent, br-broad. J inclic~tes the NMR coupling constant measured in Hertz. Continuous wave infrared (IR) spectra were recorded on a Perkin-Elmer 683 infrared spectrometer, and Fourier transform infrared (FTIR) spectra were recorded on a Nicolet Impact 400 D infrared spectrometer. IR and FTIR spectra were recorded in trAn~mi~ion mode, and band positions are reported in inverse wavenumbers (cm-l). Mass spectra were taken oneither VG 70 FE, PE Syx API III, or VG ZAB HF instruments, using fast atom bombardment (FAB) or ele~ v~L~ldy (ES) ionization techniques. Elemental analyseswere obtained using a Perkin-Elmer 240C elemental analyzer. Melting points were taken on a Thomas-Hoover melting point apparatus and are uncorrected. All te~ ela~ules are reported in degrees Celsius.
~nAltt~ch Silica Gel GF and E. Merck Silica Gel 60 F-254 thin layer plates were used for thin layer chromatography. Both flash and gravity chromatography were carried out on E. Me~ck Kieselgel 60 (230-400 mesh) silica gel. Analytical and preparative HPLC were carried out on Rainin or Berkm~n chromatographs. ODS
refers to an octadecylsilyl derivatized silica gel chromatographic support. 5 ,u Apex-ODS in~lic~ s an octadecylsilyl derivatized silica gel chromatographic support having a nominal particle size of 5 ,u, made by Jones Chromatography, Littleton,Colorado. YMC ODS-AQ~ is an ODS chromatographic support and is a registered tra(1~mArk of YMC Co. Ltd., Kyoto, Japan. PRP-l~) is a polymeric (styrene-divinylbenzene) chromatographic support, and is a registered trademark of T~milton Co., Reno, Nevada) Celite(~) is a filter aid composed of acid-washed diatomaceous silica, and is a registered trAtl~m~rk of Manville Corp., Denver, Colorado.
Methyl (+)-7-carboxy4-methyl-3-oxo-2,3,4,5-tetrahydro- lH- 1,4-- 35 benzodi~epine-2-acetate, methyl (2S)-7-carboxy-4-methyl-3-oxo-2,3,4,5-tetrahydro- 1 H- 1,4-benzodiazepine-2-acetate, methyl (2R)-7-carboxy-~methyl-3-oxo-2,3,4,5-tetrahydro-lH-1,4-ben7o~ 7~pine-2-acetate, methyl (+)-7-carboxy-4-isopropyl-3-oxo-2,3,4,5-tetrahydro- 1 H- 1,4-benzodiazepine-2-acetate, methyl (+~-7-carboxy-3-oxo-2-(2-phenylethyl)-2,3,4,5-tetrahydro- 1 H- 1,4-benzodiazepine-2-acetate, methyl (+)-8-carboxy-2-methyl-3-oxo-2,3,4,5-tetrahydro-lH-2-benzazepine-4-acetate, methyl (+) 7-amino-5-oxo-~(2-phenylethyl)-lH-1,4-5 benzodiazepine-2-acetic acid, and tert-butyl 4-fluoro-3-methylbenzoate were prepared by the method of Bondinell, et al., WO 93/00095. Methyl (~)-7-carboxy-4-(2-methoxyethyl)-3-oxo-2,3,4,5-tetrahydro- 1 H- 1,4-benzodiazepine-2-acetate, methyl (~)-7-carboxy-4-[2-(3,4-methylenedioxyphenyl)ethyl] -3-oxo-2,3,4,5-tetrahydro- 1 H-1,4-benzodiazepine-2-acet~tf- m~thyl (+)-7-carboxy-3-oxo-2,3,4,5-tetrahydro-1~I-1,4-benzodiazepine-2-acetate, methyl (+)-2,3,4,5-tetrahydro-7-[[[(ben7imi~l~7Ql-2-yl)methyl~amino]carbonyl]-4-methyl-3-oxo-lH-1,4-benzodiazepine-2-acetate, (S)-2,3,4,5-tetrahydro-7-t[[(bf~n7:imi-1~7.ol-2-yl)methyl]methylamino]carbonyl]~
methyl-3-oxo-lH-1,4-benzodiazepine-2-acetic acid, 2-(methylaminomethyl)ben~imic~7ole dihydrochloride, and 4-aza-5-rnethyl-2-(methylamino)methylben7imif~70le were prepared according to P50256-1.
Preparation 1 Preparation of 2-(aminomethyl)-4-~a-5-methylben7.imi-1~7--1e dihydrochloride a) 2,3-Diamino-6-methylpyridine 10% Pd/C (3.2 g, 3 mmole) was added to a solution of 2-Amino-6-methyl-3-nitropyridine (2.30 g, 15 mmole) in absolute EtOH (150 mL), and the mixture was shaken at RT under H2 (50 psi). After 1.5 hr, the mixture was filtered through celite~), and the filtrate was concentrated under vacuum to afford the title compound as a yellow oil. This was used without further purification: 'H NMR (250 MHz, CD30D) ~ 6.82 (d, lH), 6.36 (d, IH), 2.25 (s, 3H).
b) 2-Amino-3-[(benzyloxycarbonyl)glycyl~amino-6-nlethylL)ylidine DCC (3.09 g, 15 mmole) was added to a solution of 2,3-diamino-6-methylpyridine (15 mmole) and Cbz-glycine (3.14 g, 15 mmole) in DMF (19 mL) and CH2Cl2 (19 mL) at 0~C under argon. When the DCC had dissolved, the slightly cloudy solution was warrned to RT. After 18.5 hr, the rnixture was filtered through celite(~, and the filtrate was concentrated to dryness on the rotavap. The residue was reconcentrated from xylenes (to remove DM~) to leave a yellow solid. Silica gel chromatography (10% MeOH/CHCl3) gave the title compound (2.24 g, 48%) as a yellow solid: TLC Rf (10% MeOH/CHCl3) 0.57; 'H NMR (250 MHz, DMSO - d6) o W O 97/24119 PCTrUS96/20748 9.11 (br s, 1 H), 7.48 - 7.60 (br t, 1 H), 7.20 - 7.48 (m, 6 H), 6.40 (d, 1 H), 5.69 (br s, 2 H), 5.06 (s, 2 H), 3.82 (d, 2 H), 2.23 (s, 3 H).
c) 4-Aza-2-(benzyloxycarbonyl)aminomethyl-5-methylbenzimidazole S A solution of 2-amino-3-[(benzyloxycarbonyl)glycyl]amino-6-melhyl~ylidine (2.24 g, 7.13 mmole) in glacial AcOH (70 mL) was heated to refluxunder argon. After 17 hr, the solution was concentrated (rotavap, high vacuum), and the residue was reconcentrated from toluene (to remove AcOH). The r~slllting yellow oil was treated with hot EtOAc (20 mL) and the ll~i~lUlG was cooled to RT.
The solid was collected by suction filtration and washed with EeOAc to afford the title compound (1.72 g, 81%) as an off-white solid: TLC R, (15% MeOH/CHCl3) 0.63; MS (ES) m/e 297.4 (M + H)+.
d) 2-(Arninomethyl)-4-aza-5-methylben7imi-l~7ole dihydrochloride 10% Pd/C (153 mg, 0.14 mmole) was added to a solution of 4-aza-2-(benzyloxycarbonyl)aminomethyl-5-methylben7imi(1~7Ole (213.4 mg, 0.72 mmole) and 1.0 N HCl (1.44 mL, 1.44 mmole) in absolute EtOH (7.2 mL). The llli~lult; was purged with H2, then was stirred briskly at RT under H2 (balloon). After 2 hr, the reaction was filtered through celite, and the filtrate was concentrated on the rotavap to leave the title compound as an off-white solid: MS (ES) m/e 163.2 (M + H)+.
rr~l)aidlion 2 Preparation of methyl (+)-2.3~4.5-tetrahydro-7-carboxy-4-~3.3-dimethylbutyl)-3-oxo- lH- 1.4-ben7.odiazepine-2-acetate a) tert-Butyl 3-[(3,3-dimethylbutyl)amino]methyl-4-nitrobenzoate A ~ u~e of tert-butyl 3-methyl-4-nitrobenzoate (WO 93/00095; 17.7 g, 74.7 mmol), NBS (19.9 g, 112.0 mmol), benzoyl peroxide (1.81 g, 7.47 mmol), and CC14 (370 mL) was heated at reflux. After 17.5 h, the reaction was cooled thoroughly in ice and filtered to remove the pleci~ilaled succinimicle. The filtrate was concentrated to leave a brownish-yellow oil.
This oil ~4.2 g, 13.29 mmol), was dissolved in dry THF (50 mL), and 3,3-dimethylbutylamine (3.0 g, 29.64 mmol) was added all at once. The orangish-yellow solution was stirred at RT for 80 min, then was concentrated to remove the THF. The residue was diluted with Et20 (150 mL) and washed sequentially with 1.0 N NaOH (25 mL) and H2O (25 mL). The combined aqueous layers were back-W O 97/24119 PCT~US96120748 extracted with Et2O (50 mL~, and the combined organic layers were washed with brine (25 mL) and dried (MgS04). Concentration gave the crude title compound (4.19 g, 94%~ as a light-brown oil: MS (ES) rn/e 337.2 (M+H)+.
S b) tert-Butyl 3-[[N-(3,3-dimethylbutyl)-N-(tert-butoxycarbonyl)]amino]methyl-4-nitrobenzoate Di-tert-butyl dicarbonate (4.0 g, 18.39 mmol) was added all at once to a solution of tert-butyl 3-[(3,3-dimethylbutyl)amino]methyl-4-nitrobenzoate (4.12 g, 12.26 mmol) in CHCl3 (80 mL) at RT. After 18 h, the reaction was concen~lated and reconcentrated from hexanes (to remove CHCl3). Silica gel chromatography (10-25% EtOAc/hexanes) gave the title compound (5.0 g, 93%) as a yellow oil: MS
(ES) m/e 437.2 (M+H)+, 459.2 (M+Na)+.
c) tert-Butyl 4-amino-3-[~N-(3,3-dimethylbutyl)-N-(tert-butoxycarbonyl)]amino]methyl benzoate 10% Pd/C (1.0 g, 0.94 mmol) was added to a solution of tert-butyl 3-[[N-(3,3-dimethylbutyl)-N-(tert-butoxycarbonyl)]amino]methyl~-nitrobenzoate (4.95 g,11.35 mmol) in EtOAc (50 mL), and the mixture was shaken on a Parr apparatus at RT under H2 (55 psi). After 4 h, the reaction was filtered through celite~, and the filtrate was concentrated to afford the title compound (4.3 g, 93%) as a reddish-brown oil: MS (ES) m/e 407.4 (M+H)+.
d) tert-Butyl (~t)-4-[2-(1,4-dimethoxy-1,4-dioxobutyl)amino~-3-[[N-(3,3-dimethylbutyl)-N-(tert-butoxycarbonyl)]amino]methylbenzoate A solution of tert-butyl 4-amino-3-[[N-(3,3-dimethylbutyl)-N-(tert-butoxycarbonyl)]arnino]methylbenzoate (5.6 g, 13.79 mmol) and dimethylacetylene dicarboxylate (1.86 mL, 15.17 mmol) in MeOH (28 mL) was heated at reflux for 1 h, then was cooled to RT. The rçslllting solution was combined with MeOH (80 mL) and 10% Pd/C (2.9 g, 2.76 mmol), and the mixture was shaken on a Parr apparatus at RT under H2 (50 psi). After 22 h, the reaction was filtered throughcelite~), and the filtrate was concentrated on the rotavap. The residue was reconcentrated from CHC13 (to remove MeOH), then was chromatographed on silica gel (25% EtOAc/hexanes). The title compound (2.64 g, 42%) was obtained as a faintly yellow oil: MS (ES) m/e 551.2 (M+H)+.
e) Methyl (+)-2,3,4,5-tetrahydro-7-carboxy-4-(3,3-dimethylbutyl)-3-oxo-lH-1,4-benzodiazepine-2-acetic acid PCTrUS96120748 TFA (25 mL) was added all at once to a solution of tert-butyl (+)4-~2-(1 ,4-~iimPth~xy-1 ,4-dioxobutyl)amino]-3-~[N-(3,3-dimethylbutyl)-N-(tert-butoxycarbonyl)]amino]methylbenzoate (12.64 g, 4.8 mmol) in anhydrous CH2Cl2 (25 mL) at 0~C, and the faintly yellow solution was warmed to RT. After 1 h, thesolution was concenLlaLed on the rotavap, and the residue was reconcentrated from toluene (to remove residual TFA). The resulting oil was combined with toluene (50 mL) and Et3N (3.34 mL, 24 mmol), and the mixture was heated to reflux. A light yellow, homogeneous solution was produced. After 16 h, the reaction was concentrated on the rotavap to leave a solid residue. This was dissolved in a minimllm of MeOH (ca. 10 mL), diluted with H2O (10 mL), and acidif1ed with glacial AcOH to pH 4.5. The ~ lulc was filtered, and the precipitate was washed sequentially with MeOH and E~t20, then was dried in high vacuum to afford the title compound (1.88 g, 93%) as a nearly colorless powder: MS (ES) m/e 363.2 (M+H)+.
~lGI)aldlion 3 Preparation of Bisr(ben7imi-1~7ol-2-yl)methyllamine tris(trifluoroacetate) a) Bis[[1-N-(tert-butoxycarbonyl)bçn7imi(1~7Ol-2-yl]methyl]-N-(tert-butoxycarbonyi)amine To a stirred solution of 2-aminomethylbenzimidazole dihydrochloride hydrate (6.26 g, 28.4 mmol) and triethylamine (4.0 mL, 28.4 mmol) in dry THF (50mL) was added l-(tert-butoxycarbonyl)-2-(bromomethyl)bçn7imit1~7c 1e (P50256-1;
2.00 g, 9.48 mmol) in THF (30 mL),. After 8 h, a solution of di-tert-butyl dicarbonate (10.0 g, 45.84 mmol~ in CHC13 (50 mL) was added slowly. The resulting mixture was stirred at RT overnight then was concentrated. The residuewas taken up in CH2C12 (150 mT.), and washed sequentially with water (60 rnL), 5%
NaHCO3 (60 mL), and brine (60 mL). Drying (MgSO4), concentration, and silica gel chromatography (6% MeOH/CH2C12) gave the title compound (0.46 g, 8%) as a faint yellow oil: MS (ES) m/e 578.4 (M+H)+.
b) Bis[(bPn7imic1:~701-2-yl)methyl]amine tris(trifluoroacetate) A solution of TFA (3 mL) and CH2C12 (9 mL) at RT was added all at once to bis[[1 -N-(tert-butoxycarbonyl)benzimidazol-2-yl]methyl}-N-(tert-butoxycarbonyl)amine (0.23 g, 0.4 mmol). After 35 min, the solution was concentrated on the rotavap, and the residue was reconcentrated from toluene (to 107 W O 97/24119 PCTrUS96~0748 remove residual TFA) to afford the title compound (0.17 g, 68%) as an off-white powder: MS (ES) m~e 278.0 ~M+H)+.
Preparation 4 Preparation of 2-rrl-r(benzimidazol-2-yl)methyllben7imi(1~7ole~methyllamine bi~(Jri lluorQacetate) a) [[l-N-(tert-Butoxycarbonyl)be~7imi-1~701-2-yl3methyl]-N-(tert-butoxycarbonyl)amine To a stirred solution of 2-aminomethylben7imidzl701e dihydrochloride hydrate (3.0 g, 13.63 mmol) and triethylamine (8.44 rnL, 61.3 mmol) in dry CH~C12 (50 mL) a solution of di-tert-butyl dicarbonate (6.54 g, 30.0 mmol) in CH2Cl2 (50 rnL) was added at 0~C. The reaction was stirred at RT for 1 h, then more of triethylamine (1.9 mL, 13.8 mmol) and di-tert-butyI dicarbonate (2.97 g, 13.63 mmol) were added. The reslllting mixture was stirred at RT for 24 h, then was concenkated. The residue was taken up in CH2Clz (50 mL) and washed sequentially with 0.5 N HCl (2x40 mL), 5% NaHC03 (50 mL), and brine (50 mL). The crude product was recrys~lli7ecl from CH2Cl~/ether to give the title compound (2.8 g, 59%) as a wnite powder: MS (ES) m/e 348.2 (M~H)+.
b) 2-[[1-[[( 1-(tert-Butoxycarbonyl)ben7imirl~701-2-yl3methyl]ben7.imitl~7.01e]methyl]-N,N-di-(teLt-butoxycarbonyl)amine To a stirred solution of [[l-N-(tert-butoxycarbonyl)bPn7imirl~7Ql-2-yl]methyl]-N-(di-tert-butoxycarbonyl)amine (0.6 g, 1.73 mmol) and NaH (0.1 g, 4.17 mmol) in dry THF (12 mL) and DMF (4 mL) was added 1-(tert-butoxycarbonyl)-2-(bromomethyl)ben7imi~ 701e ~0.6 g, 1.93 nimol). Theresulting Lul~ was stirred at RT for 1 h, then was concenLIdL~d. The residue was taken up in CH2C12 (100 mL) and washed sequentially with water (50 mL), 5% NaHC03 (30 mT ), and brine (30 mL). Drying (MgS04), concentration, and silica gel chromatography (2:3 EtOAc/hexanes) gave the title compound (0.27 g, 27%) as a faint yellow oil: MS (ES) m/e 578.2 (M~H)+.
c) 2-~[l-[(Ben7imi~1~7-)l-2-yl)methyl]bçn7imitl~7~1e~methyl]amine bis(trifluoro~et~1r) A solution of TFA/CH2C12 (30 ml, 25%) at RT was added all at once to 2-[[1-[[( 1 -N-tert-butoxycarbonyl)bel-7 i ~ 701-2-yl]methyl]ben7imi~1~701e]methyl]-N,N-CA 0224l633 l998-06-26 W O 97/24119 PCT~US96/20748 di-(tert-butoxycarbonyl)amine (0.25 g, 0.43 mmol). After 25 min, the solution was concentrated on the rotavap, and the crude product was recryst~ d from CH2Cll/ether to give the title co~ oulld (0.17 g, 63%) as an off-white powder:
MS (ES) m/e 278.0 ~M+H)+.
~xamI)le 1 a~dLion of (+)-2.3~4.5-tetrahydro-7-rrr(4-~a-5-methylbellzi.ll~dazol-2-yl)methyllaminolcarbonyll -4-(2-methoxyethyl)-3-oxo- 1 H- 1.4-benzodiazepine-2-a~tic acid a) Methyl (+)-2,3,4,5-tetrahydro-7-[[[(4-aza-5-methylben7imiclzl7Ql-2-yl)methyl]amino]carbonyl]~(2-methoxyethyl)-3 -oxo- 1 H- 1,4-benzodiazepine-2-acetate EDC (138 mg, 0.72 mrnole) was added to a solution of methyl (+)-7-carboxy-4-(2-methoxyethyl)-3-oxo-2,3,4,5-tetrahydro- lH- 1,4-benzodiazepine-2-acetate (202 mg, 0.60 mmole), 2-(arninomethyl)-4-aza-5-methylben7imicl~7Ole dihydrochloride (0.72 mmole), HOBt ~ H2O (97 mg, 0.72 mmole), and diisopropylethylamine (0.84 mL, 4.8 mmole) in anhydrous CH3CN (3 mL) at RT.
After 16 hr, the reaction was concentrated, and the residue was reconcellllaL~d from xylenes/CHCl3. Silica gel chromatography (15% MeOH/CHCI3) gave the title compound (impure): TLC Rf (15% MeOH/CHCI3) 0.55; MS (ES) m/e 481.5 (M +
H)t. This was used without further purification.
b) (+)-2,3,4,5-Tetrahydro-7-[tt(4-aza-S-methylben7imi~1~7ol-2-yl)methyl]amino~carbonyll-4-(2-methoxyethyl)-3-oxo- 1 H- 1,4-benzodiazepine-2-acetic acid A two-phase mixture of methyl ( )-2,3,4,5-tetrahydro-7-[[[(4-aza-5-methylbel.~.i...ic1sl7.Ql-2-yl)methyl]amino]carbonyl]-4-(2-methoxyethyl)-3-oxo-lH-1,4-benzodiazepine-2-acetate (0.60 mrnole), 1.0 N LiOH (1.8 mL, 1.8 mmole), and THF (4.2 mL) was stirred at RT for 45 min, then was concentrated to remove the THF. The aqueous layer was washed with Et2O (2 x 2 mL), and the Et2O layers werediscarded. The aqueous layer was diluted with CH3CN (2 mL) and acidified with TFA (0.23 mL). The res-llting solution was concentrated to dryness on the rotavap, and the residue was purified by ODS chromatography (12~o CH3CN/H20 contS~ini 0.1% TFA (250 mL), then 15% CH3CN/H20 cont:~ining 0.1% TFA). Concentration and lyophilization gave the title compound (199.5 mg, 50% for two steps) as a light W O 97/24119 PCT~US96/20748 yellow powder: HPLC (PRP~ , 15% CH3CN/H20 cont~ining 0.1%
TFA) K' = 1.4; MS (ES) rn/e 467 (M + H)'. Anal. Calcd for C23El26N6Os ~ 1.5 CF3COzH 1.33 H2O: C, 47.21; H, 4.60; N, 12.70. Found: C, 47.20; H, 4.73; N, 12.79.
S .
Examl~le 2 Pl~a~tion of (_)-2~3~4~s-tetr~ydro-7-rrr(ben7im~ 7~ol-2-yl)methyllaminolcarbonyll-4-(2-methoxyetl~ -3-oxo-lH-1.4-ben7.o-lJa7epine-2-acetic acid a) Methyl (_)-2,3,4,5-tetrahydro-7-[[t(bel ,7i . l . if 1~7f~l-2-yl)methyl]arnino]carbonyl]
4-(2-methoxyethyl)-3-oxo- lH- 1 ,4-benzodiazepine-2-acetate EDC (230 mg, 1.2 mmole) was added to a solution of methyl (+)-7-carboxy-4-(2-methoxyethyl)-3-oxo-2,3,4,5-tetrahydro- lH- 1 ,4-benzodiazepine-2-acetate (336.4 mg, 1.0 mmole), 2-(aminomethyl)be~7imif1~7f-1e dihydrochloride hydrate (264 mg, 1.2 mmole), HOBt ~ H2O (162 mg, 1.2 mmole), and diisopropylethylamine (0.70 mL, 4.0 mmole) in anhydrous DMF (5 mL) at RT. After 17 hr, the reaction was concentrated, and the residue was reconcentrated from xylenes (2 x) to remove DMF. The residue was diluted with H2O (3 mL) and extracted with CHCl3 (3 x 5 mL). The combined extracts were treated with MeOH (2 rnL) to dissolve a precipitate, then were dried (MgSO,) and concentrated. Reconcentration from xylenes (to remove residual DMF) left a light yellow solid. This was dissolved in MeOH/CHCI3, and the solution was concentrated to leave an oil. Silica gel chromatography (10% MeOH/CHCl3) gave an off-white solid, which was triturated with EtOAc (3 mL) to afford the title compound (397.1 mg, 85%) as a colorless solid: TLC R, (10% MeOH/CHCl3) 0.46; MS (ES) m/e 466.2 (M + H)~.
b) (_)-2,3,4,5-Tetrahydro-7-[~[(ben7imi(1~7Ol-2-yl)methyl]amino]carbonyl]-4-(2-methoxyethyl)-3-oxo- lH- 1 ,4-benzodiazepine-2-acetic acid 1.0 N LiOH ( 1.0 mL, 1.0 mmole) was added to a suspension of methyl (+)-2,3,4,5-tetrahydro-7-[[[~benzimidazol-2-yl)methyl3amino]carbonyl]~(2-methoxyethyl)-3-oxo-lH-1,4-ben7o~ 7f-pine-2-acetate (397 mg, 0.85 mmole) in THF (4.3 rnL) and H20 (3.3 mr ) at RT. The light yellow mixture was stirred at 40 -50~C for 1 hr, and the reslllting homogeneous solution was then stirred at RT for 17.5 hr. The reaction was concentrated, and the rçslllting oil was dissolved in H20 (4 mL). The solution was filtered to remove particnl~tes, and the filtrate was CA 0224l633 l998-06-26 W O97/24119 PCTrUS96/20748 neutralized with 1.0 N HCl (1.0 rnL). The yellowish solid was collected and triturated with good stirring with hot 1: 1 CH3CN/H2O. The resulting solid was collected, washed with plenty of 1: 1 CH3CN/H2O, and dried in high vacuum (40~C)to afford the title compound (327.9 mg, 85%) as a colorless powder: HPLC (PRP-S l(E~), 15% CH3CN/H20 cont:~inin~ 0.1% TFA) K' = 4.6; MS (ES) m/e 452.2 (M +
H)~. Anah Calcd for C23H25N5O5: C, 61.19; H, 5.58; N, 15.51. Found: C, 61.18; H,5.58; N, 15.39.
~ExamE)le 3 Preparation of (+)-4-~4-~rr(lH-~en7~imicla7Ql-2-yl)methyllmethylaminolcarbonyllphenyll-3-phenylbutanoic acid a) Ethyl 3-hydroxy-4-(4-methoxyphenyl)-3-phenylbutanoate Anhydrous EtOAc (4.3 mL, 44 mmole) was added dropwise over 5 - 6 rnin to a solution of lithium bis(trimethylsilyl)amide (1.0 M in THF, 40 mL, 40 mmole) in dry THF (60 mL) in a flame-dried flask at -78~C under argon. The yellow solution was stirred at -78~C for 0.5 hr, then a solution of 2-(4-methoxyphenyl)-1-phenyleth~n<)nP (Chem. Ber. 195S, 91, 755-759; 4.53 g, 20 mmole) in dry THF (20 ml) was added dropwise over 12 rnin. Additional THF (2 mL) was used in transfer.After 0.5 hr, The reaction was quenched with satd NH4Cl (120 mL) and warmed to RT. EtOAc extraction, drying (MgSO4), concentration, and silica gel chromatography (20% EtOAc/hexanes) gave the title compound (6.13 g, 96%) as a light yellow oil: TLC Rf (20% EtOAc/hexanes) 0.34; MS (ES) m/e 315.2 (M + H)~.
b) Ethyl 4-(4-methoxyphenyl)-3-phenylbutanoate Boron trifluoride etherate (4.8 mL, 39 mmole) was added dropwise over 3 rnin to a solution of ethyl 3-hydroxy-4-(4-methoxyphenyl)-3-phenylbutanoate (6.13 g, 19.5 mmole) and triethylsilane (6.2 mT, 39 mmole) in anhydrous CH2C12 (49 rnL) at 0~C under argon. The reaction was stirred at RT overnight, then was quenched with 5% NaHCO3 (100 mL). The ll~i~lul~ was stirred briskly for 10 min, then was separated. The aqueous layer was extracted with CH2Cl2 (100 mL), and the combined organic layers were dried (Na2SO4) and concentrated. The residue was reconcentrated from hexanes (to remove CH2Cl2) to leave a yellow oil. This was - 35 dissolved in absolute EtOH (100 mT.), and 10% Pd/C (775 mg, 1.95 mmole) was added. The mixture was shaken on a Parr apparatus at RT under H2 (50 psi) for 2 hr, then was filtered through celite~3). The filtrate was concentrated, and the residue was W O 97/24119 PCT~US96/20748 chromatographed on silica gel (15 % EtOAc/hf~x~n~s). The title compound (5.27 g, 91%) was obtained as a colorless oil: TLC Rr (15% EtOAc/hexanes) 0.40;
MS (ES) m/e 299.2 (M + H)+.
c) Ethyl 4-(4-hydroxyphenyl)-3-phenylbutanoate Anhydrous ~ minllm trichloride (4.49 g, 33.7 mmole) was added all at once to solution of ethyl 4-(4-methoxyphenyl)-3-phenylbutanoate (2.01 g, 6.74 mmole) and eth~nethiol (2.5 mT., 33.7 mrnole) in anhydrous CH2Cl2 (67 mL) at 0~C under argon. The yellow solution was warmed to RT and stirred for 3 hr, then was recooled to 0~C and ~luen~hl cl with cold 3 N HCl (67 mL). The mixture was stirred for 5 rnin, then was separated. The aqueous layer was extracted with CH2Cl2 (2 x100 mL), and the combined organic layers were dried (Na2SO4) and concentrated.
Silica gel chromatography (25% EtOAc/hexanes) gave the title compound (1.84 g, 96%) as a colorless oil: TLC Rf (30% EtOAc/hexanes) 0.47; MS (ES) m/e 285.2 (M
+ H)'.
d) Ethyl 3-phenyl-4-[4-(trifluoromethanesulfonyloxy)phenyl]butanoate Trifluorom~th~n.oslllfonic anhydride (1.4 mL, 8.4 mrnole) was added rapidly -dropwise to a solution of ethyl 4-(4-hydroxyphenyl)-3-phenylbutanoate (1.84 g, 6.47 rnmole) and 2,6-lutidine (1.5 mL, 12.9 mmole) in anhydrous CH2Cl2 (32 mL) at -78~C under argon. After 0.5 hr, the yellow solution was warmed to RT and stirredfor 1 hr. The reaction was diluted with Et2O (150 mL) and washed sequentially with 1.0 N HCl (15 mL), 5% NaHCO3 (15 mL), and saturated brine(15 mL). Drying (MgS04), concentration, and silica gel chromatography (15% EtOAc/hexanes) gave the title compound (2.62 g, 97%) as a nearly colorless oil: TLC Rf (20%
EtOAc/hexanes) 0.55; MS (ES) m/e 417.0 (M + H)~.
e) Ethyl 4-(4-carboxyphenyl)-3-phenylbllt:~n~-~t~
A mixture of ethyl 3-phenyl~-[4-~trifluornm~th:-nesulfonyloxy)phenyl]butanoate (2.62 g, 6.29 mmole), anhydrous KOAc (2.47 g, 25.16 mmole), Pd(OAc)2 (70.6 mg, 0.31 mrnole), dppf (697.4 mg, 1.26 mmole), and anhydrous DMSO (31 mL) was purged with carbon monoxide (three evacuation/ carbon monoxide purge cycles, followed by bubbling carbon monoxide through the mixture for 5 min), then was heated at 70~C under a balloonof carbon monoxide. After 3.5 hr, the reaction was diluted with H2O (31 mL), cooled in ice, and acidified with 1.0 N HCl (25 mL). CH2C12 extraction (2 x 100 mL), drying (MgSO4), concentration, and reconcentration from toluene left a :
W O 97/24119 PCT~US96/20748 reddish-orange liquid. Silica gel chromatography (1% AcOH in 7:3 toluene/EtOAc) gave the title compound (1.78 g, 91 %) as a cream-colored solid:
TLC Rf (1% AcOH in 7:3 toluene/EtOAc) 0.47; MS (ES) m/e 313.2 (M + H)+.
S f) Ethyl (+)-4-[4-[~[(lH-bell~hllidazol-2-yl)methyl]methylamino~carbonyllphenyl]-3 -phenylbutanoate - EDC (230 mg, 1.2 mmole) was added to a solution of ethyl 4-(4-carboxyphenyl)-3-phenylbutanoate (312.4 mg, 1.0 rnrnole), 2-(methylaminomethyl)ben7imi(1z-7ole~dihydrochloride (281 mg, 1.2 mmole), HOBt lQ H2O (162 mg, 1.2 rnmole), and diisopropylethylamine (0.70 mL, 4.0 mmole) in anhydrous CH3CN (5 mL) at RT. After 18 hr, the reaction was concentrated, and the brown residue was chromatographed on silica gel (5% MeOH in 1: 1 EtOAc/CHCl3).
The title compound (439.2 mg, 96%) was obtained as a light orange foam: TLC Rf (5% MeOH in 1: 1 EtOAc/CHCl3) 0.50; MS (ES) m/e 456.2 (M + H)+.
g) (~)~[4-[[[(}H-Benzimidazol-2-yl)methyl]methylamino]carbonyl]phenyl]-3-phenylbutanoic acid A solution of ethyl (_)-4-[4-[[[(lH-ben7imitl~7ol-2-yl)methyl]methylamino]carbonyl]phenyl~-3-phenylbutanoate (439.2 mg, 0.96 20 mmole) and 1.0 N NaOH (1.2 mL, 1.2 mmole) in EtOH (8.4 mL) was stirred at 50~C. After 24 hr, the reaction was concentrated to dryness and the residue was purified by ODS chromatography (35% MeOH/H20). Concentration and lyophilization gave the title compound (412.2 mg, 86%) as a colorless powder:
HPLC (PRP-l(~), 35% CH3CN/H20 cont~inin~ 0.1% TFA) K' = 1.4; MS (ES) 428 25 (M + H)', 450 (M + Na)~. Anal. Calcd for C26H2JN3O3Na 2.75 H2O: C, 62.58; H, 5.96; N, 8.42. Found: C, 62.34; H, 5.84; N, 8.44.
Rx~m,l?le 4 30 ~le~)a dlion of (+~-4-14-rrr(ben7imi~1~7~-1-2-yl)methyllmethylaminolcarbonyllphenyll-3-(dimethylaminocarbonyl)butanoic acid a) tert-Butyl 4-bromobenzoate Trifluoromethanesulfonic acid (0.18 rnL, 2 rnmole) was added dropwise to a - 35 mixture of 4-bromobenzoic acid (20.10 g, 100 rnrnole), anhydrous CH2Cl2 (100 mL), and condensed isobutylene (-78~C; 100 mL), and the rf s~ in~ mixture was allowedto reflux under a dry ice/acetone condenser. After 40 min, more isobutylene (30 CA 0224l633 l998-06-26 W O 97/24119 rcTrusg6/2o748 mL) was added, and reflux was continued for an additional 20 min. The reaction was poured into Et2O (500 mL) and washed sequentially with 1.0 N KOH (2x 50 rnL), H2O (50 mL), and satd brine (50 mL). Drying (MgSO4), concentration, and silica gel chromatography (5% EtOAc/hexanes) gave the title compound (15.28 g, 59%) as a light yellow oil: TLC Rf (5% EtOAc/hexanes) 0.59; MS (ES) m/e 259/257 (M + H)+.
b) Methyl 3-[4-(tert-buto~y~;all~onyl)phenyl]propenoate A solution of tert-butyl 4-hromobenzoate (5.14 g, 20 mmole), methyl - 10 acrylate (9.1 rnL, 100 mrnole), Pd(OAc)2 (224.5 mg, 1 rnrnole), tri-o-tolylphosphine (608.8 mg, 2 rnmole), and diisopropylethylamine (7.0 mL, 40 mmole) in propionitrile (100 rnL) was heated at reflux for 3 hr, then was concentrated on the rotavap. The residue was diluted with Et2O (200 mL) and washed sequentially with1.0 N HCl (2 x 50 mT), 5% NaHCO3 (50 rnL), and satd brine (50 mL). Drying (MgSO4), concentration, and silica gel chromatography (15% EtOAc/hexanes) gave the title compound (3.34 g, 64%) as a light yellow solid: TLC Rf (20%
EtOAc/hexanes) 0.51; MS (ES) rn/e 263.0 (M + H)+.
c) Methyl 3-[4-(tert-butoxycarbonyl)phenyl]propanoate 10% Pd/C (2.71 g, 2.55 mrnole) was added to a solution of methyl 3-[4-(tert-butoxycarbonyl)phenyl]propenoate (3.34 g, 12.73 mmole) in EtOAc (65 rnL) and MeOH (65 rnL), and the mixture was shaken on a Parr a~paldLus at RT under H2 (50psi). After 3 hr, the reaction was filtered through celite~, and the rlltrate was concentrated to dryness on the rotavap. Reconcentration from hexanes left the title compound (3.27 g, 97%) as a cloudy, grayish oil: TLC Rf (20% EtOAc/hexanes) 0.63; MS (ES) m/e 265.0 (M + H)+.
d) 3-t4-(tert-Butoxycarbonyl)phenyl]propanoic acid A rnixture of methyl 3-[4-(tert-butoxycarbonyl)phenyl]propanoate (3.27 g, 12.37 rnrnole), 1.0 N LiOH (14.8 mL, 14.8 mmole), THF (31 mL), and H2O (16 mL) was stirred at RT for 1.5 hr, then was concentrated on the rotavap to remove theTHF. The aqueous solution was washed with Et20 ~2 x 30 rnL), and the Et,O layerswere discarded. The aqueous layer was acidified with 1.0 N HCl (ca. 17 mL), and the mixture was extracted with CHCl3 (3 x 50 mL). Drying (Na2SO4) and concent;ation gave the title compound (3.04 g, 98%) as a colorless powder: mp 88.5 - 89.5~C; MS (DCVNH3) m/e 268.0 (M ~ NH4)+.
W O 97/24119 PCT~US96/20748 e) N,N-Dimethyl 3-[4-(tert-butoxycarbonyl)phenyl]prop~n~mi~l EDC (2.09 g, 10.88 mmole) was added to a solution of 3-[4-(tert-butoxycarbonyl)phenyl]propanoic acid (2.27 g, 9.07 mmole), di~llelllylamine hydrochloride (0.88 g, 10.88 mmole), HOBt ~ H2O (1.47 g, 10.88 mmole), and diisopropylethylamine (3.2 rnL, 18.14 mmole) in anhydrous CH3CN (45 mL) at RT.
After 19.5 hr, the reaction was concentrated, and the residue was chromatographed on silica gel (EtOAc). The title compound (2.46 g, 98%) was obtained as a colorless oil: TLC Rf (EtOAc) 0.52; MS (ES) mte 278.4 (M + H)~.
f) Ethyl 4-[4-(tert-butoxycarbonyl)phenyl]-3-(dimethylaminocarbonyl)butanoate A solution of lithium bis(trimethylsilyl)amide in THF (1.0 M, 5.8 mL, 5.8 mmole) was added dropwise over 2.5 min to a solution of N,N-dimethyl 3-[4-(tert-butoxycarbonyl)phenyl]prop~n~mi~lf~ (1.34 g, 4.83 mmole) in dry THF (48 mL) at -78~C under argon. The yellow solution was stirred at -78~C for 0.5 hr, then ethyl bromoacetate (2.7 mL, 24.15 mmole) was added over 15 sec down the walls of the flask (to precool). After 0.5 hr, the reaction was poured into satd NH4Cl (50 rnL), and the mixture was extracted with EtOAc (2 x 100 mL). Drying (MgSO4), concentration, and reconcentration from xylenes left a light yellow oil. Silica gel chromatography (1: 1 EtOAc/hexanes) gave the title compound (453.5 mg, 26%) as alight yellow oil: TLC Rf (1: 1 EtOAc/hexanes) 0.44, MS (ES) m/e 364.2 (M + H)+.
g) Ethyl 4-(4-carboxyphenyl)-3-(dimethylaminocarbonyl)b~lt:~noz-tf~
TFA (2.3 mL) was added all at once to a solution ethyl 4-[4-(tert-butoxycarbonyl)phenyl] 3-(dimethylaminocarbonyl)butanoate (168.6 mg, 0.46 mmole) in anhydrous CH2Cl2 (2.3 rnL) at 0C. The solution was stirred at RT for 0.5 hr, then was concentrated to dryness on the rotavap. The residue was reconcentrated from toluene to afford the title colllpoulld as a light yellow oil: MS (ES) mle 308.0 (M + H)1.
h) Ethyl (+)-4-[4-t[[(ben7 i ~ I .ifl~7ol-2-yl)methyl]methylamino~call,onyl]phenyl}-3-(dimethylaminocarbonyl)bl~t~ o~t~
EDC (105.8 mg, 0.55 mmole) was added to a solution of ethyl 4-(4-carboxyphenyl)-3-(dimethylaminocarbonyl)butanoate (0.46 mmole), 2-(methylaminomethyl)ben7imicl~7ole dihydrochloride (129.2 mg, 0.55 mmole), - 35 HOBt ~ H2O (74.6 mg, 0.55 mmole), and diisopropylethylamine (0.32 mL, 1.84 mmole) in anhydrous CH3CN (2.3 mL) at RT. After 22 hr, the reaction was concentrated, and the yellow residue was chromatographed on silica gel (10%
W O 97/24119 PCTrUS96/20748 MeOH in 1:1 EtOAc/CHCl3). The title compound (191.5 mg, 92%) was obtained as a light yellow oil: TLC Rf (10% MeOH in 1:1 EtOAc/CHCl3) 0.44; MS
(ES) m/e 451 (M + H)+.
S i) (V-4-[4-[r[(l~en7.imicl~7.ol-2-yl)methyl]methylamino]carbonyl]phenyl]-3-~dimethylaminocarbonyl)butanoic acid A solution of ethyl (i)-4-[4-[[[(benzirnidazol-2-yl)methyl]methylamino]carbonyl]phenyl]-3-(dimethylaminocarbonyl)butanoate (191.5 mg, 0.43 mmole) and 1.0 N LiOH (0.52 mL, 0.52 mmole) in T~IF (2.2 mL) and H~O (1.6 mL) was stirred at RT for 17 hr, then was acidified with TFA (0.10 rnL, 1.29 mmole). Concentration left an aqueous residue which was purified by ODS chromatography (17~o CH3CN/H2O cont:~inin~ 0.1% TFA; chromatographed againusingl5%CH3CN/H2Ocont~ining0.1%TFA). Concentrationand lyophilization gave the title compound (133.4 mg, 4791'o) as a colorless powder:HPLC (PRP-1(~), 20% CH3CN/H20 cont:lining 0.1% TFA) K' = 1.3; MS (ES) m/e 423.2 (M + H)+. Anal. Calcd for C23H26N,O4 ~ 2 CF3CO2H ~ 0.5 H2O: C, 49.17; H, 4.43; N, 8.49. Found: C, 49.13; H, 4.62; N, 8.52.
T~x~ ?le S
Preparation of (S)-2.3.4.5-tetrahydro-7-rrr(ben7imi~7ol-2-yl)mPthyllmethylaminolcarbonyll-4-mPthyl-3-oxo-lH-1 .~ben70diazepine-2-acetic :~rid. r(2.2--limethyl-2-methoxyacetyl)oxy~methyl ester a) (S)-2,3,4,5-Tetrahydro-7-[r[[1-(tert-butoxycarbonyl)bc~ 7Q1-2-yl]methyl]methylamino]carbonyl]-4-methyl-3 -oxo- 1 H- 1 ,4-benzodiazepine-2-acetic acid To a ~ Lulc of (S)-2,3,4,5-tetrahydro-7-r[[(ben7imi~ 7ol-2-yl)methyl~methylamino]carbonyl3-4-methyl-3-oxo- I H- l ,4-benzodiazepine-2-acetic acid (444 mg, 1.0 mmol), triethylamine (0.1464 mT.,l.OS mmol) in DMF (8 mL), was added dropwise di-tert-butyl dicarbonate (230 mg, 1.05 mmol) in DMF (2 mL).
The reaction ll~i~lule was stirred at RT for 18 h. An aliquot was assayed to inrii~ ~te.
only 50% conversion. Another quantity of triethylamine and di-tert-butyl dicarbonate were added and stirring was continued for another 18 h. An aliquot still in-liC~tefl some unreacted material, so a third quantity of reagents was added and the reaction was stirred for another 18 h. The reaction mixture was concentrated to dryness and the residual oil was triturated with water, filtered and vacuum dried at W O 97/24119 PCTrUS96/20748 40-50~C, to give a white solid of the title compound ( 0.442 g, 85%). MS
fES) m/e 522.4 ~M+H]+.
b) (S)-2,3,4,5-Tetrahydro-7-[[l[l-(tert-butoxycarbonyl)ben7imi~l~7ol-2-yl]methyl]methylamino]carbonyl]-4-methyl-3-oxo- lH- 1,4-benzodiazepine-2-acetic acid, ~(2,2-dimethyl-2-methoxyacetyl)oxy]methyl ester To a solution of the compound of Example Sa, (0.209 g, 0.4 mmol) in dIy acetone (10 mL) was added anhydrous polassiulll carbonate (0.25 g, 1.8 mmol). The reaction mixture was stirred at RT and under argon for 1 h. 2-methoxy-2-methylpropanoic acid chloromethyl ester, US Patent 4,602,012, July 22, 1986, (0.334 g, 2.0 mmol) was then added, followed by tetrabutylammonium iodide (0.03 g, 0.08 mmol). The reaction was stirred at RT under argon for 48 h. It was then filtered and the filtrate was concentrated to a yellow oily residue of the titlecompound (0.67g, q~nti~ative yield). TLC RF 0.48 (silica gel, 6% methanol in methylene chloride). MS ~ES) m~e 652.2 [M+H]+.
c) (S)-2,3,4,5-Tetrahydro-7-[[[(ben7imicl~7ol-2-yl)methyl]methylamino]carbonyl]-4-methyl-3 -oxo- 1 H- 1,4-benzodiazepine-2-acetic acid, [(2,2-dimethyl-2-methoxyacetyl)oxy]methyl ester To a solution of the compound in Example Sb (0.67 g, 1 mmol) in methylene chloride (5 mL) was added TFA (1 mL). The reaction was stirred at RT under argonfor 4 h. It was concentrated to dryness, and the residue was evaporated with methylene chloride three times to remove TFA traces, to give the title compound (0.4 g, 73%). This was purified on a flash silica column (step gradient, 2-3%
methanol in methylene chloride). The fractions cont:linin~ the pure compound were collected, concentrated to yield the title compound (65 mg ) as an off-white solid.
MS (ES) m/e 552.2 [M+H]+. 'H NMR (400 MHz, (CDCl3) ~ 7.6 (br s, lH), 7.22 (m, 6H), 6.5 ( d, lH), 5.85 ( d, lH), 5.8 (d, lH), 5.4 (d, lH), 5.05 (m, lH), 4.79 (q, 2H), 4.3 (d, lH), 3.7 (d, lH), 3.25 ( s, 3H), 3.15 (s, 3H), 3.05 ( s, 3H), 3.02 ( dd, lH), 2.7 (dd, lH), 1.4 (s, 6H). Anal. Calcd for C~8H33N507 1.25 H20: C, 58.58; H, 6.23; N, 12.20. Found: C, 58.60, H, 5.94, N, 12.00.
Fxam~?le 6 P.~paldlion of fi)-2.3.4.5-tetrahydro-7-rrrfben~imi~ 7Ql-2-yl)methyl~ aminolcarbonyll-4-methyl-3-oxo- 1 H- 1.4-benzodiazepine-2-fN-hydroxy~ fzlmi~l~
W O 97124119 PCT~US96/20748 a) (~)-2,3,4,5-Tetrahydl o-7-[[~(benzimidazole-2-yl)methylamino]call,ollyl]-4-methyl-3-oxo- 1 H- 1,4-benzodiazepine-2-(N-hydroxy)~cetz~
NaOMe (Aldrich, 25 wt. % solution in MeOH, 2.2 mL, 9.7 mmole) was added to a solution of hydroxylamine hydrochloride (0.67 g, 9.7 mmole) in MeOH
(40 mT-) at 45~C, and the ~ Lul~e was stirred for 5 min. Methyl (~-2,3,4,5-tekahydro-7-[[[(kpn7imi~ Ql-2-yl)methyl]amino]carbonyl]-4-methyl-3-oxo-lH-1,4-benzodiazepine-2-acetate (0.82 g, 1.9 mmole) was s-lcpçn~lP-l in MeOH (2 mL)and THF (15 mL) and added dropwise to the above solution. The reaction was then stirred at 45~C for 24 h. The ll~ lul~ was concentrated in vacuo, and was then treated with 10% CH3CN/H20 conl~inin~ 0.1% TFA (5 mL). All material dissolved, and then a solid precipitated out. Half of this material was dissolved in the mobile phase by addition of excess TFA and was purified by. prepd.d~ e HPLC (YMC
ODS-AQ, 50 x 250 mm, flow rate = 80 mL/min, 10% CH3CN/H20 con~z~ining 0.1%
T~A; tR = 57 min) to yield the title compound (91 mg, 22%) as a white solid. MS
(ES) m/e 423.1 [M+H] '. 'H NMR (400 MHz, DMSO-d6) ~ 9.06 (bt, J = 4 Hz, lH), 7.77 (m, 2H), 7.58 (m, 2H), 7.50 (m, 2H), 6.60 (d, J = 10 Hz, lH), 6.40 (bs, lH), 5.52(d,J= l9Hz, IH),5.18(bt,J=9Hz, lH),4.85(d,J=6Hz, lH),3.83(d,J=
19 Hz, lH), 2.95 (s, 3H), 2.60 (dd, J = 17, 9 Hz, lH), 2.28 (dd, J = 15, 7 Hz, IH).
Anal. Calcd for C2,H22N6O" ~ 1.5 C2HF3O2 1.0 H2O): C, 47.14; H, 4.20; N, 13.74.
Found: C, 46.95; H, 4.24; N, 13.37.
F.x~nu?le 7 Preparation of (+)-2.3.4~5-tetrahydro-7-r3-(benzimidazol-2-yl~pher~vll-4-methyl-3-oxo- 1 H- 1 ~4-benzodiazepine-2-acetic acid a) 2-(3-Iodophenyl)benzilllidazole To a cold solution of 3-iodobenzoic acid (5~0 g, 20 mmol) and Et3N (3~7 mL, 26 mmol) in THF (50 mL) was added isobutylchloroformate (2~9 mL, 21 mmol3.
The solution was stirred for 1 h at 10~C All of the solution was added slowly to a solution of 1,2-diaminobenzene (2~2 g, 20 mmol) in THF (50 mL)~ After 18 hr, thereaction was concentrated and the residue was partitioned between EtOAc and 5%
Na2CO3~ The layers were separated and the EtOAc layer was washed with water.
Concentration of the organic layer gave a residue which was treated with EtOAc and allowed to stand for 1~ min. Filtration gave a solid which was treated with AcOH(50 rnL) and heated to 110~C After 18 hr, the solution was concentrated. The W O 97/24119 PCTrUS96/20748 residue was treated with EtOAc and the solution was filtered to give the title cc,~ oulld (3.14 g, 50%): MS (ES) m/e 321.2 (M+H)~.
b) 2-[(3-Tributylstannyl)phenyl]l,en,;...icl~7Ole S A solution of 2-(3-iodophenyl)ben7imitl~7Ole (1.0 g, 3.1 mmol), bistributyltin (3.9 mL, 6.2 mmol) and PdCl~(PPh3)2 (100 mg, 0.14 rnmol) in DMF
(10 mL) was heated to 90~C under argon. After 2 hr, the solution was concentrated.
The residue was treated with hexane and filtered. EtOAc was added and the solution was filtered. The filtrate was concentrated to give the title compound (812 mg, 54%): MS (ES) m/e 485.4 (M+H)'.
c) Methyl (i)-2,3,4,5-tetrahydro-1-(tert-butoxycarbonyl)-7-iodo-4-methyl-3-oxo-1 H- 1,4-benzodiazepine-2-acetate To a solution of methyl-7-iodo-4-methyl-3-oxo-lH-1,4-benzodiazepine-2-acetate (1.6 g, 4.3 mmol) and DMAP (10 mg, 0.08 mmol) in CH3CN (10 mL) was added di-tert-butyl dicarbonate (2.0 g, 8.6 mmol), and the solution was stirred at RT.
Additional di-tert-butyl dicarbonate (a total of 8 g, 34.4 mmol) was added periodically until the reaction went to completion. Concentration and silica gelch~ lugraphy gave the title compound (1.8 g, 90%): MS (ES) mle 497.2 (M+Na)' d) Methyl (i)-2,3,4,5-tetrahydro-7-[3-(ben7imi~1~7ol-2-yl)phenyl]- 1 -(tert-butoxycarbonyl)-4-methyl-3-oxo- lH- 1,4-benzodiazepine-2-acetate A mixture of 2-[(3-tributylstannyl)phenyl3ben7imicl~7ole (0.24 g, 0.5 mmol), methyl (+)- 1 -(tert-butoxycarbonyl)-7-iodo-4-methyl-3-oxo- 1 H- 1,4-benzodiazepine-2-acetate (0.223 g, 0.7 mmol), CuI (10 mg, 0.05 mmol), and PdCl2(PPh3)z (40 mg, 0.05 mmol) in DMF (10 mL) was heated to 100~C under argon. After 18 hr, the solution was concentrated. Silica gel chromatography gave the' title compound (0.06 g, 22%): MS (ES) m/e 541.5 (M+H)t.
e) Methyl (_)-2,3,4,5-tetrahydro-7-[3-(be~.~i.. ifl~7ol-2-yl)phenyl]-4-methyl-3-oxo-lH- 1,4-benzodiazepine-2-acetate A solution of methyl (+)-2,3,4,5-tetrahydro-7-[3-(ben7imi~1~7~ 1-2-yl)phenyl]-1 -(tert-butoxycarbonyl)-4-methyl-3-oxo- 1 H- 1,4-benzodiazepine-2-acetate (0.06 g, 0.11 mmol) in 4 M HCl/dioxane (3 mL) was stirred for 1 hr at RT. The solution was concentrated to give the title compound (0.05 g, 100%): MS (ES) m/e 441.4 (M+H)~.
W O 97/24119 PCT~US96/20748 f) (_)-2,3,4,5-Tekahydro-7-[3-(be~ idazol-2-yl)phenyl]-4-methyl-3-oxo-lH-1,4-ben7O~ 7~pine-2-acetic acid 1.0 N NaOH (0.22 mL, 0.22 mmol) was added dropwise to a solution of methyl (+)-2,3,4,5-tetrahydro-7-[3-(b~n7imill~7ol-2-yl)phenyl]-4-methyl-3-oxo-lH-1,4-benzodiazepine-2-acetate (0.05 g, 0.11 mmol) in 1: 1 MeOH/H2O (2 mL) at RT, and the resulting solution was stirred for 18 hr, then was concentrated. The residue was dissolved in H,O and the solution was acidified with AcOH to pH 4 (Litmus paper). Filtration gave the title compound (0.005 g, 10%): 'H NMR (250 MHz, DMSO-d6) o 2.4-2.9 (m, 2H), 3.0-3.1 (s, 3H), 3.8-4.0 (d, lH), 5.0-5.1 (m, lH), 5.5-5.6 (d, lH), 6.7-6.8 (d, lM), 7.5-8.5 (m, 1 lH); MS (ES) m/e 427.5 (M+H)f. Anal.Calcd for C25H"N4O3 1.5 HCl - 1.0 AcOH 0.5 H20: C, 58.94; H, 5.22; N, 10.18.
Found: C, 59.00; H, 5.15; N, 9.92.
Fx~rr~le 8 P1~AI ~lion of (+)-2.3.4.5-tetrahydro-4-methyl-3-oxo-7-r~rtph~ n~ l ., hnidazol-2-yl)m~ thyll"minolcarbonyll- 1 H- 1.4-benzodiazepine-2-acetic acid a) 2-[[(N-Benzyloxycarbonyl)amino]methyl]phen~nthrimi~1~7Ole Following the general procedure of Exarnple 7(a), except substituting N-Cbz-glycine for the 3-iodobenzoic acid, and 9,10-tliz~minophen~nthrene for the 1,2-diaminoben_ene, the title compound (0.41 g, 45%) was prepared: MS (ES) m/e 382.4 (M+H)i.
b) 2-(Aminomethyl)phen"nthrimi(1~7Ole A solution of 2-t[(N-benzyloxycarbonyl)amino~methyl]ph~ ~nthrimidazole (0.2 g, 0.52 mmol) in 30% HBr in acetic acid (0.8 mL) was stirred at RT for 1 hr.
The solution was concentrated and the residue was treated with Et2O. Filtration gave the title compound (0.138 g, 80%) as an oily residue: MS (ES) m/e 248.3 (M+H)~.
c) Methyl (_)-2,3,4,5-tetrahydro-4-methyl-3-oxo-7-[[~(phen~nthrimi~z~7ol-2-yl)methyl]amino]carbonyl]- lH- 1,4-benzodiazepine-2-acetate EDC (0.08 g, 0.42 mmol) was added to a solution of 2-(aminomethyl)ph~n:~nthrimi~ le (0.138 g, 0.42 rnmol), methyl ( )-7-carboxy~
methyl-3-oxo-2,3,4,5-tetrahydro-lH-1,4-benzodiazepine-2-acetate (0.123 g, 0.42 mmol), HOBt H~O (0.063 g, 0.42 mmol) and Et,N (0.14 mL, l mmol) in anhydrous ~20 W O 97/24119 PCTrUS96/20748 DMF (5 rnL~ at RT. After 18 hr, the reaction was concentrated, and the residue was partitioned between EtOAc and 5% NaHCO3. The layers were separated and the organic layer was washed with H2O. Drying (NazSO4) and concentration gave the title compound (0.2 g, 90%): MS (ES) m/e 522.4 (M+H)+.
d) (+)-2~3~4~5-Tetrahydro-4-methyl-3-oxo-7-[[[(phpn~nthrim~ 7ol-2 - yl)methyl]arnino]carbonyl]-lH-1,4-bçn7o~ 7~pine-2-acetic acid Following the procedure of Example 7(f), methyl (+)-2,3,4,5-tetrahydro-4-methyl-3-oxo-7-[[[(phen~nthrimi(lzl71>1-2-yl)methyl]amino}carbonyl]-lH-1,4-benzodiazepine-2-acetate (0.2 g, 0.38 mmol) was saponified and purified to give the title compound (0.014 g, 10%): MS (ES) m/e 508.5 (M+H)+. Anal. Calcd for C29H25N5O4 1.0 TFA 3.0 H2O: C, 55.11; H, 4.77; N, 10.37. Found: C, 55.38; H, 5.13; N, 10.74.
F.x~nl~l?le 9 Ple~)a dlion of methyl (+)-7-carboxy-4-(2.2~2-trifluoroethyl)-3-oxo-2.3.4.5-tetr:~ydro- 1 H- 1.4-benzodiazepine-2-acetate a) tert-Butyl 3-[(2,2,2-trifluoroethyl)arnino]methyl-4-nitroben7O~t~
tert-Butyl 3-bromomethyl-4-nitrobenzoate (2.4 g, 8 mmol)was dissolved in dry THF (50 mL), and 2,2,2-trifluoroethylamine (3 rnL, 38 mmol) was added all atonce. The orangish-yellow solution was stirred at RT for 40 min, then was concentrated to remove the TH~. The residue was diluted with Et20 (100 rnL) and washed twice with 10 % aqueous Na2CO3 (50 mL) and brine (50 rnL). The organic layer was dried (MgSO4). Concentration and silica gel chromatography (2.5% -10% EtOAc/hexanes) gave the title compound (1.6 g, 63%) as a yellow oil: IH
NMR (250 MHz, CDC13) o 8.21 (d, J = 1.3 Hz, lH), 8.03 (dd, J = 8.4, 1.3 Hz, lH),7.96 (d, J = 8.4 Hz, lH), 4.20 (s, 2H), 3.24( q, J = 9.3 E~z, 2H), 1.62 (s, 9H).
b) tert-Butyl 3-[[N-(2,2,2-trifluoroethyl)-N-(tert-buto~yc~l~onyl)3amino]methyl-4-nitro~en7O~te Di-tert-butyl dicarbonate (2.15 g, 10 mmol) was added all at once to a solution of tert-butyl 3-[(2,2,2-trifluoroethyl)amino]methyl-4-nitrobenzoate (1.6 g, 5 mmol) in CH2Cl2 (25 mT.) at RT. The reaction was concentrated and heated to 50~Cunder vacuum for 18 hours. Silica gel chromatography (2% - 5% EtOAc/h~-x~n~s) gave the title compound (2 g, 96%) as a yellow oil: ~H NMR (400 MH_, CDCl3) W O 97/24119 PCTrUS96/20748 7.85-8.15 (m, 3H), 4.75-5.05 (m, 2H), 3.80-4.10 (m, 2H), 1.60 (s, 9H), 1.15-1.80 (m, 9H).
c) tert-Butyl 4-amino-3-[[N-(2,2,2-trifluoroethyl)-N-(tert-butoxycarbonyl)]amino~methylben70~tl ~
10% Pd/C (.4 g, .4 mmol) was added to a solution of tert-butyl 3-[[N-(2,2,2-trifluoroethyl)-N-(tert-butoxycarbonyl)3amino~methyl-4-nitrobenzoate (2.0 g, 5 mmol) in EtOAc (20 mL), and the ~ Lul~ was shaken on a Parr apparatus at RT
under H2 (55 psi). After 4 h, the reaction was filtered through celite(~), and the filtrate was concentrated to afford the title compound (1.9 g, 99%) as a colorless oil:
lH NMR (400 MHz, CDC13) ~ 7.76 (dd, J = 8.5 Hz, 1.8 Hz, lH), 7.68 (d, J = 1.8 Hz, lH), 6.62 (d, J = 8.4 Hz, lH), 4.53 (s, 2H), 3.69 (m, 2H), 1.58 (s, 9H), 1.51(m, 9H).
d) tert-Butyl (+)-4-[2-(1,4-dimethoxy- 1,4-dioxobutyl)anuno]-3-[[N-(2,2,2-trifluoroethyl~-N-(tert-butoxycarbonyl)]amino]methylbenzoate A solution of tert-butyl 4-amino-3-[[N-(2,2,2-trifluoroethyl)-N-(tert-butoxycarbonyl)]amino]methylbenzoate (1.9 g, S mmol) and dimethylacetylene dicarboxylate (0.58 mL, S.S mmol) in MeOH (10 mL) was heated at reflux for 60 rnin, then was cooled to RT. The resulting solution was combined with MeOE~ (20 mL) and 10% Pd/C (0.5 g, .S mrnol), and the mixture was shaken on a Parr apparatus at RT under H2 (50 psi). After 3 h, the reaction was filtered through celite~, and the filtrate was concentrated on the rotavap. The title compound (1.6 g, 62%) was obtained as a faintly yellow oil: IH NMR (40V MHz, CDC13) ~ 7.85 (dd, J = 8.4, 2.0 Hz, lH), 7.68 (d, J = 2.0 Hz, lH), 6.65 (d, J = 8.4 Hz, lH), 6.15 (br s, lH), 4.55~.70 (m, 2H), 4.40 (1/2 AB, J = 15.3 Hz, lH), 3.71 (s, 3H), 3.70 (s, 3H), 3.35-3.50 (m, 2H), 2.95 (dd, J = 16.9, 6.8 Hz, lH), 2.84 (dd, J = 16.9, 6.9 Hz, lH), 1.56 (s, 18H).
e) Methyl (+)-7-carboxy-4-(2,2,2-trifluoroethyl)-3-oxo-2,3,4,5-tetrahydro-lH-1,4-benzodiazepine-2-acetate TFA (7 mL) was added all at once to a solution of tert-butyl (+)-4-[2-(1,4-~lim~thoxy- 1,4-dioxobutyl)amino}-3-[[N-(2,2,2-trifluoroethyl)-N-(tert-butoxycarbonyl)]amino]methylbenzoate (1.6 g, 3 mmol) in anhydrous CH2C12 (20 rnL) at 0~C, and the faintly yellow solution was warmed to RT. After 2 h, the solution was concentrated on the rotavap, and the residue was reconcentrated from toluene (to remove residual TFA). The reSlllting oil was combined with toluene (10 mL) and Et3N (2 mL, lS mmol), and the mixture was heated to reflux under argon.
After 18 hr the solution was allowed to cool and was concenll~ted to dryness under W O 97/24119 PCT~US96120748 vacuum. The residue was dissolved in a miniml~m of MeOH (ca. 15 mL) at reflux, diluted with H2O ~10 ml), and acidified with glacial AcOH (4 drops). The mixture was kept in the refrigerator overnight then was filtered. The solid was dried under high vacuum to afford the title compound (0.80 g, 76%) as a tan powder: lHNMR (400 MHz, DMSO-d6) o 7.61 (2, lH), 7.57 ~dd, J = 8.5, 2 Hz, lH), 6.63 (d, J =
2 Hz, lH), 6.59 (d, J = 8.5 Hz, lH), 5.59 (d, ~ = 16.7 Hz, lH), 5.25 (m, lH), 4.28 (m, 2H), 4.15 (d, J = 16.7 Hz, lH), 3.61 (s, 3H), 2.86 (dd, J = 16.8, 8.7 Hz, lH), 2.74 (dd, J = 16.8, 5.4 Hz, lH).
f) Methyl (+)-2,3,4,5-tetrahydro-7-[[~(benzimidazol-2-yl)methyl]amino]carbonyl]-3-oxo-4-(2,2,2-trifluoroethyl)- 1 H- 1 ,4-bçn70~ 7epine-2-acetate EDC (0.16 g, 0.86 mmol) was added at RT to a solution of methyl (+)-7-carboxy4-(2,2,2-trifluoroethyl)-3-oxo-2,3 ,4,5-tetrahydro- 1 H- 1 ,4-benzodiazepine-2-acetate (0.20 g, 0.71 mmol), HOBt ~ H2O (0.12 g, 0.86 mmol), 2-aminomethylb~n7imill~701e dihydrochloride (0.19 g, 0.86 mmol), DIEA (0.5 mL, 2.8 mmol), and acetonitrile (5 mL) under argon. The rçslllting solution was stirred at RT overnight, then was concentrated. The residue was partitioned between ethyl acetate and water, and the layers were separated. The organic phase was washed with brine, dried (MgSO4), and concentrated. Silica gel chromatography ~1% - 10%CH30H in CH2Cl~) gave the title compound (0.12 g, 44%) as a tan solid: NMR (400 MHz, DMSO-d6) o 8.59 (t, J = 5 Hz, lH), 7.61 (m, 2H), 7.50 (m, 2H), 7.16 (m, 2H), 6.57 (d, J = 11.1 Hz, lH), 6.17 (d, J = 5 Hz, lH), 5.53 (d, J = 16.7 Hz, lH), 5.13 (m, lH), 4.75 (m, 2H), 4.10 (m, 2H), 3.62 (s, 3H), 2.94 (dd, J = 16.8, 8.5 Hz, lH), 2.69 (dd, J = 16.8, 5.4 Hz, lH).
g) (+)-2,3,4,5-Tetrahydro-7-[[[(bçn7imi~1~7ol-2-yl)methyl]amino~carbonyl]-3-oxo-4-(2,2,2-trifluoroethyl)- 1 H- 1 ,4-benzodiazepine-2-acetic acid A solution f methyl(f)-2,3,4,5-tetrahydro-7-[[[(bçn7imi~l~7ol-2-yl)methyl]amino]carbonyl]-3-oxo4-(2,2,2-trifluoroethyl)- lH- 1 ,4-ben7o~ 7~pine-2-acetate (0.12 g, 0.25 mmol) and lithium hydroxide monohydrate (0.017 g, 0.4 mmol) in THF (10 mL), CH30H (2 mL), and H2O (2 mL) was stirred at RT overnight. It was then concentrated and the residue was dissolved in water. The solution was brought to pH 4 with 3 N HCI, then was refrigerated for 1 hour. The resulting solid was collected by filtration and dried to give the title co~ oul-d (0.11 g, 90%) as a ~ 35 white solid: Ms (ES) m/e 476 ~M+H]~. Anal. Calcd for C22H2oN5F304 1.25 H20: C, 53.07; H, 4.55; N, 14.06. Found: C, 52.85; H, 4.36; N, 13.98.
W O 97/24119 PCT~US96/20748 Example 10 Preparation of f+)-2.3.4.5-tetr~ dro-7-rrrbenzimidazol-2-yl)methyllmethylaminolcarbon,yll-3-oxo-4-(2.2.2-trifluoroethyl)- 1 H- 1.4-S berl7~ 7Ppine-2-acetic acid a) Methyl (+)-2,3,4,5-tetrahydro-7-[[[ben7imi(1:-7ol-2-yl)methyl]methylamino]carbonyl~-3-oxo-4-(2,2,2-trifluoroethyl)- I H- 1,4-benzodiazepine-2-acetate Using the procedures of Example 9(f), except substituting 2-(methylaminomethyl)bçn7imic~7ole dihydrochloride for the 2-aminomethylbenzimidazole dihydrochloride, the title compound was prepared: 'H
NMR (400 MHz, CDCl3) ~ 7.67 f~m, 2 H), 7.37 (m, 2 H), 7.25 (m, 2 H), 6.54 (d, J =
8 Hz, 1 H), 5.46 (d, J = 16.7 Hz, 1 H), 5.20 (m, 1 H), 5.04 (s, 2 H), 4.71 (m, 1 H), 4.17(m,1H),3.94(m,1H),3.92(d,J=16.7,1H),3.74(s,3H),3.23(s,3H),2.98 (m, 1 H), 2.74 (m, 1 H).
b) (i)-2,3,4,5-Tetrahydro-7-[trbçn7imiti~7ol-2-yl)methyl]methylamino]carbonyl]- -3-oxo-4-(2,2,2-trifluoroethyl)- 1 H- 1,4-benzodiazepine-2-acetic acid Using the procedure of Example 9(g), methyl (+)-2,3,4,5-tetrahydro-7-[[rbenzimidazol-2-yl)methyl]methylamino]carbonyl] -3-oxo-4-(2,2,2-trifluoroethyl)-lH-1,4-benzodiazepine-2-acetate was saponified to afford the title compound: MS
f~ES) mte 490.2 [M+H~+. Anal. Calcd for C23H22N5F30" 2.25 H20: C, 52.12; H, 5.04; N,13.21. Found: C, 52.00; H, 5.12; N, 13.09.
Fx~n~l?le 11 Prep ~ration of (+)-2.3.4.5-tetrahydro-7-rrr(4-aza-S-methylben7.imicl~7ol-2-yl~rnf~tllyllaminolcarbonyll-3-oxo-4-f2.2.2-trifluoroethyl)-lH-1.4-benzodiazepin~-2-zlf~Ptic acid a) Methyl (+)-2,3,4,5-tetrahydro-7-~[r(4-aza-5-methylben7imirl:~7OI-2-yl)methyl]amino]carbonyl]-3-oxo-4-(2,2,2-trifluoroethyl)- 1 H- 1,4-benzodiazepine-2-acetate Using the procedures of Fx:~rnpl~ 9(f~, except ~ubsLiLuLing 2-(aminomethyl)-4-aza-5-methylben7imi~ 7O1e dihydrochloride for the 2-~minom( thylben7imi-l~7oledihydrochloride, the title colllp-)ul~d was prepared: MS (ES) rnte 505.2 ~M + H)+.
W O97124119 PCTrUS96/20748 b) (~ 2,3,4,5-Tetrahydro-7-[[[(4-aza-5-methylben7imi~1~7ol-2-yl)methyl]amino}carbonyl]-3-oxo-4-(2,2,2-trifluoroethyl)- lH- 1,4-b~4n7.o-1iA7epine-2-acetic acid Using the procedure of Example 9(g), methyl (~)-2,3,4,5-tetrahydro-7-[[[(4-aza-5-methylbenzimidazol-2-yl)methyl]amino]carbonyl]-3-oxo-4-(2,2,2-~ trifluoroethyl)-lH-1,4-benzodia~epine-2-acetate was saponified to afford the title compound: MS (ES) m/e 491.2 (M+H)+. Anal. Calcd for C~H2lN6F3O4 - 2 7/8 H2O:
C, 48.73; H,4.97; N, 15.50. Found: C, 48.50; H, 4.59; N, 15.33.
F.x~n~le 12 al~lion of (+)-2.3.4.5-tetrahydro-7-rrr(ber~7imi(1~7ol-2-yl)methyllmethylamino]carbonyll-4-r2-(3.4-methylenedioxyphenyl)ethyl]-3-oxo-lH-1.4-benzotli~7~-pine-2-acetic acid a) Methyl (+)-2,3,4,5-tetrahydro-7-[[[(ben7imi~ 7~-1-2-yl)methyl3methylamino]carbonyl]-4-[2-(3,4-methylenedioxyphenyl)ethyl]-3-oxo-lH- 1,4-benzodiazepine-2-acetate EDC (0.10 g, 0.55 mmol) was added at RT to a solution of methyl 7-carboxy-4-[2-(3,4-methylenedioxyphenyl)ethyl]-3-oxo- 1 H- 1,4-benzodi~epine-2-acetate (0.14 g, 0.3 mmol), 2-(methylaminomethyl)ben7imicl:~701e dihydrochloride(0.12 g, 0.51 mmole), HOBt ~ H20 (0.072 g, 0.55 mmol), and DIEA ~0.32 mL, 1.84 mmole), in DMF (5 mL) under argon. The resulting solution was stirred at RT
overnight, then was concentrated. The residue was partitioned between ethyl acetate and water, and the layers were separated. The aqueous phase was extracted with ethyl acetate, and the combined organic phases were washed with brine, dried (MgSO4), and concentrated. Silica gel chromatography gave the title compound (0.11 g, 59%) as a colorless foam: 'H NMR (CDCl 3) o 7.62 (m, 2H), 7.31 (m, 2H),7.20 (d, J = 8.1 Hz, lH), 7.07 (s, lH), 6.65 (d, J = 7.9 Hz, lH), 6.60 (s, lH), 6.55 (d, J=7.9Hz, lH),6.46(d,J=8.1 Hz, lH),5.90(d,J=5.4Hz,2H),5.26(d,J= 16.5 Hz, lH), 5.02 (m, lH), 4.93, (d, J = 14.6, lH), 4.83 (d, J = 14.6 Hz, lH), 4.51 (d, J =
5 Hz, lH), 3.74 (s, 3H), 3.71 (m, lH), 3.60 (m, lH), 3.58 (d, J = 16.5 Hz, lH), 3.18 (s, 3H), 2.99 (dd, J = 16, 6.8 Hz, lH), 2.70 (m, lH).
b) (+)-2,3,4,5-Tetrahydro-7-[[[(ben7imi~1~7Ol-2-yl)methyl]methylamino]carbonyl]-4-~2-(3,4-methylenedioxyphenyl)ethyl] -3-oxo- 1 H- 1,4-benzodiazepine-2-acetic acid W O97/24119 PCT~US96/20748 To a solution stirred at RT of methyl (_)-2,3,4,5-tetrahydro-7-[[[(benzirnidazol-2-yl)methyl]methylamino]carbonyl]-4-[2-(3,4-methylenedioxyphenyl)ethyl]-3-oxo-lH-1,4-benzodiazepine-2-acetate (0.11 g, 0.19 rnmol) in THF (1 mL) was added a solution of lithium hydroxide monohydrate (0.01g, 0.23 rnrnol) in H20 (1 mL). The r~s~ ing solution was stirred at RT overnight, then was concentrated to dryness. The residue was dissolved in H2O and the solution was washed with ethyl acetate, then was brought to pH 4 with 3 N HCl.
The res~ ing precipitate was collected by ~lltration and dried to give the titlecompound (0.055 g, 51%) as a white solid. MS (ES) m/e 556.2 [M+H]+. Anal.
Calcd for C3nH29NsO6 H2O: C, 62.82; H, 5.45; N, 12.21. Found: C, 62.69; H, 5.26;N, 12.15.
Fx~ ?le 13 ~lel)a-~lion of (+)-2.3.4.5-tetrahydro-7-rl2-(benzimidazol-2-yl)acetyll~mino]-5-oxo-4-(2-phenylethyl)- 1 H- 1.4-benzodi~epine-2-acetic acid a) Methyl (+)-2,3,4,5-tetrahydro-7-[t2-(be~ idazol-2-yl)acetyl]amino]-5-oxo-4-(2-phenylethyl)- 1 H- 1,4-benzodiazepine-2-acetate EDC (0.27 g, 1.4 rnmole) was added at RT to a solution of (_)-2,3,4,5-tetrahydro-7-amino-5-oxo4-(2-phenylethyl)- 1 H- 1,4-benzodiazepine-2-acetate (0.40 g, 1.1 mmol), ben7imi~1~7Qle-2-acetic acid (Archiv. Der Pharmazie 1960, 293, 758;
0.25 g, 1.4 rnmol), HOBt ~ H[2O (0.20 g, 1.5 mmol), and DIEA (0.35 mL, 2 mmol) in acetonitrile (10 rnL). The res~ ing solution was stirred for 2 days, then was concentrated to dryness. The residue was partitioned between ethyl acetate and water, and the layers were separated. The organic phase was washed with brine, dried (MgSO4), and concentrated. Silica gel chromatography (1% - 10% CH30H in CH2Cll) gave the title compound (0.21 g, 36%) as an amber foam: MS (ES) m/e 512.2 [M+H]+.
b) (+)-2,3,4,5-Tetrahydro-7-t[2-(benzimidazol-2-yl)acetyl]amino]-5-oxo4-(2-phenylethyl)- 1 H- 1,4-benzodiazepine-2-acetic acid A solution of methyl (_)-2,3,4,5-tetrahydro-7-[~2-(ben7.imi~ 1-2-yl)acetyl]amino]-5-oxo4-(2-phenylethyl)- 1 H- 1,4-benzodiazepine-2-acetate (0.21 g, 0.41 mmole) and lithium hydroxide monohydrate (0.022 g, 0.52 mmol) in THF (10 mL) and H20 (2 mL) was stirred at RT overnight, then was then concentrated. The residue was dissolved in water and the solution was washed with ethyl acetate, then W O 97124119 PCT~US96/20748 was brought to pH 4 with 3 N HCl. The resulting precipitate was collected by filtration and dried to give the title compound (0.12 g, 59%) as an off-white solid:
MS (ES) m/e 498.2 [M+H]+. Anal. Calcd for C28H27N5O4 ~ 1.5 H2O: C, 64.11; H, 5.76; N, 13.35. Found: C, 64.36; H, 5.57; N, 13.21.
Exarnple 14 Preparation of (S)-2.3~4.5-tetrahydro-7-rrr(benzimidazol-2-yl)methyllmethylaminolcarbonyl1 -4-methyl-3-oxo- 1 H- 1.4-benzodiazepine-2-acet~mi~l~
a) (S)-2,3,4,5-tetrahydro-7-t[[(bçn7imi~1~7Ol-2-yl)methyl]methylamino]carbonyl]-4-methyl-3-oxo- 1 H- 1,4-benzodiazepine-2-~et~mi~le Methyl (+)-2,3,4,5-tetrahydro-7-[[[(ben ~ 7Ol-2-yl)methyl]amino]callJonyl]-4-methyl-3-oxo-lH-1,4-benzodiazepine-2-acetate (330 mg, 0.76 mmole) in dry MeOH (10 mL) was cooled in an ice bath as ammonia was bubbled into the solution for 0.5 hr. The reaction was then allowed to sit stoppered at RT for 18 hr. After concentration, the residue was purified by silica gel flash chromatography (90: 10 CH2Cl2/MeOH) to give the title compound (52%) as a white solid: MS (ES) m/e 421.2 ~M+H~'. Anal. Calcd for C2~H24N603 1.5 H~O: C, 59.05;
H, 6.08; N, 18.78. Found: C, 58.90; H, 6.04; N, 18.45.
Fx~n~le 15 ~ aldtion of (+)-S-rr2.3.4.5-tetrahydro-7-rrr(benzimidazol-2-yl)methyllamino~carbonyll-3-oxo4-(2-phenylethyl)- 1 H- 1.4-benzodiazepin-2-yllm~thylltetrazole a) Methyl (+)-2,3,4,5-tetrahydro-7-(tert-butoxycarbonyl)-3-oxo-4-(2-phenylethyl)-lH-l,~benzodiazepine-2-acetate Methyl (+)-2,3,4,5-tetrahydro-7-carboxy-3-oxo~-(2-phenylethyl)- 1 H- 1,4-b~n70~ 7~pine-2-acetate (1.0 g, 2.6 mmole) was suspended in toluene (10 mL), andN,N-dimethylformz-micle-di-tert-butyl acetal (5 mL, 20.8 mmole) was added dropwise. The reaction mixture was heated at 80~C for 1.5 hr, then was cooled toRT and poured into 5% Na~CO3 solution. The layers were separated, and the aqueous was extracted with toluene (2 x). The combined organic layers were W O 97/24119 PCT~US96/20748 washed with brine, dried over MgS04, filtered and evaporated to give the title compound (0.9lg, 82%). MS (ES) m/e 439.2 [M+H]+.
b) (i)-2,3,4,5-Tetrahydro-7-(tert-butoxycarbonyl)-3-oxo-4-(2-phenylethyl)- lH- 1,4-benzodiazepine-2-acetic acid A solution of methyl (i)-2,3,4,5-tetrahydro-7-(tert-butoxycarbonyl)-3-oxo-4-(2-phenylethyl)-lH-1,4-benzodiazepine-2-acetate (1.5 g, 3.4 mmole) in ethylene glycol dimethyl ether (160 mL) was treated with H2O (20 mT ) and 0.91 N NaOH (5 mT.). The reaction was stirred under argon at RT for 24 hr, then was acidified to pH
3 with glacial AcOH, concentrated to a small volume (10 mL), and poured into iceH2O. The precipitated solid was collected and dried giving the title compound inqll~ntit~tive yield. MS (ES) m/e 425.2 ~M+H]+.
c) (+)-2,3,4,5-Tetrahydro-7-(tert-butoxycarbonyl)-3-oxo-4-(2-phenylethyl)- lH- 1,4-1 5 benzodiazepine-2-[N-(2-cyanoethyl)~cet~mide]
A solution of (i)-2,3,4,5-tetrahydro-7-(tert-butoxycarbonyl)-3-oxo-4-(2-phenylethyl)-lH-1,4-benzodi~epine-2-acetic acid (1.2 g, 2.6 mmole) in dry DMF
(12 mL) under argon was treated with diisopropylethylamine (0.65 g, 5 mrnole), EDC (0.764 g, 4 rnmole) and HOBt ~ H2O (0.54g, 4 mmole). The resulting solution was stirred for 10 min, then was treated with a solution of 3-aminopropionitrilefumarate in dry DMF (2 mL) cont~inin~ diisopropylethylamine (0.85 g, 6.6 lnrnole).
The reaction was stirred under argon ~or 18 hr, then was concentrated to dryness.
The residue was partitioned between H,O and EtOAc, and the layers were separated.
The organic layer was washed with brine, dried over MgSO4, filtered and concentrated. The oily residue was purified by silica gel flash chromatography (98:2 CH2Cl~/MeOH) to give the title colllpoulld (750 mg, 50%): MS (ES) 477.2 [~+H]+.
d) (i)-1-(2-Cyanoethyl)-5-[[2,3,4,5-tetrahydro-7-(tert-butoxycarbonyl)-3-oxo-4-(2-phenylethyl)- lH- 1 ,4-benzodiazepin-2-yl]methyl]tetrazole A solution of ~+)-2,3,4,5-tetrahydro-7-(tert-butoxycalbollyl)-3-oxo~-(2-phenylethyl)-lH- 1 ,4-benzodi~epine-2-[N-(2-cyanoethyl)acet~mide] (750 mg, 1.3 mmole) in dry THF (15 mL) under argon was treated with triphenylphosphine (1.14 g, 4.6 mrnole), trimethylsilyl~ide (0.52 g, 4.6 mmole), and diethyl~odicarboxylate (0.8 mL, 4.6 mmole) at RT under argon. After 50 hr, the reaction was concentrated to dryness, and the residue was purified by silica gel flash chromatography (98.5:1.5 CH2Cl2/MeOH) to give the tit~e co~ ou,ld (0.56g, 86%): MS (ES) m/e 502.2 [M+H]+.
CA 0224l633 l998-06-26 W O97/24119 PCTrUS96/20748 e) (+)-1-(2-Cyanoethyl)-5-[[2,3,4,5-tetrahydro-7-carboxy-3-oxo-4-(2-phenylethyl)-lH-1,4-benzodiazepin-2-yl]methyl]tetrazole A solution of (+)-1-(2-cyanoethyl)-5-[[2,3,4,5-tetrahydro-7-(tert-butoxycarbonyl)-3-oxo-4-(2-phenylethyl)- 1 H- 1,4-benzodi~epin-2-yl~methyl]tetrazole (0.5 g, 1 mmole) in CH2Cl2 (20 mL) at RT under argon was treated with 4 M HCl in dioxane (10 mL). After 20 hr, the reaction was concentrated to dryness, and the residue was diluted with 5% Na2CO3. The solution was extracted with EtOAc, and the ELOAc layer was discarded. The aqueous layer was acidified with dil HCl and extracted with EtOAc (3x). The combined EtOAc extracts were washed with brine, dried over MgSO4 and evaporated to yield the title compound (0.36g, 81%): MS(ES) m/e 445.4 [M+H]+.
f) (+)-1-(2-Cyanoethyl)-5-[[2,3,4,5-tetrahydro-7-[[[(b~n7imi~1~7ol-2-yl)methyl]amino]carbonyl] -3 -oxo-4-(2-phenylethyl)- 1 H- 1,4-benzodiazepin-2-yl]methyl]tetrazole A solution of (+)-1-(2-cyanoethyl)-5-[[2,3,4,5-tetrahydro-7-carboxy-3-oxo-4-(2-phenylethyl)-lH-l,~ben7ofli~7epin-2-yl]methyl]tetrazole) (360 mg, 0.8 rnmole)dry DMF (5 mL) was treated under argon with DIEA (129 mg, 1 ~nmole), EDC (172 mg, 0.9 mmole) and HOBt ~ H2O (122 mg, 0.9 mmole). The reaction was stirred at RT for 10 min, then a solution of 2-aminomethylben7imit~701e dihydrochloride hydrate (352 mg, 1.6 mmole) in DMF (2 mL) cont:lining DIEA (413 mg, 3.2 mmole) was added. After 20 hr, the reaction was concentrated to dryness and the residue was partitioned between }~2O and EtOAc. The layers were separated, and the aqueous layer was extracted with EtOAc. The organic layers were combined, dried over MgSOt, filtered and concentrated. The residue was purified by silica gel flash chromatography (95:5 CH2Cl2/MeOH) to give the title compound (120 mg, 21%):
MS (ES) m/e 575.2 [M+H]+.
g) (+)-S-[r2,3,4,5-Tetrahydro-7-[[[(b~ 7-)1-2-yl)methyl]amino]carbonyl]-3-oxo~-(2-phenylethyl)- l H- 1,4-benzodiazepin-2-yl]methyl]tetrazole A solution of (+)-1-(2-cyanoethyl)-5-[[2,3,4,5-tetrahydro-7-[[[(ben7imi~ 7Ol-2-yl)methyl]amino]carbonyl]-3-oxo-4-(2-phenylethyl)- I H- 1,4-benzodiazepin-2-yl]methyl]tetrazole (100 mg, 0.2 mmole) in MeOH (1 mL) was treated with ~ 35 thiophenol (0.02 mL) followed by 1 N NaOH solution (2.2 mL). After 3 hr, the reaction was concentrated to dryness, and the residue was purified by silica gel~pa,a~ive TLC (85: 15 CH2Cl2/MeOH). The isolated product was dissolved in HzO, W O 97/24119 PCT~US96/20748 and the solution was filtered to remove insoluble m~t~ri~l~ The filtrate was treated with 2 drops of glacial AcOH. The precipitated solid was collected and dried to give the title compound (45 mg, 41%): MS (ES) m/e 522.2 ~M+H]'. Anal.
Calcd for C3~ Ng04 ~ 2.25 H2O: C, 56.15; H, 5.52; N,19.65. Found: C, 56.51; H, 5.05; N, 19.72.
~x~rn,ple 16 Preparation of (S)-2.3.4.5-tetrahydro-7-rrrfbe~ .idazol-2-10 yl~methyllaminolcarbonyll-3-oxo4-r4-r(2-carboxybenzoyl)aminolbut-1-yll-lH-1.4-ben70~ pine-2-acetic acid a) N-[[2-(N-4-hydroxybut- 1-yl)aminomethyl-4-tert-butoxycarbonylJphenyl~-L-aspartic acid ,l3-methyl ester A mixture of N-[[2-formyl-4-tert-butoxycarbonyl]phenyl]-L-aspartic acid ,13-methyl ester (WO 95/18619; 2.55g, 7.26 mmol), 4A molecular sieves, and 4-hydroxybutylamine (0.64 g, 7.26 mmol) in MeOH (35 mL) was stirred under argon at RT for 30 min, then sodium cyanoborohydride (0.49 g, 0.79 mmol) and acetic acid (0.3 mL) were added. The reaction mixture was kept at RT overnight and then20 the solvent was elimin~te~l in vacuo. The residue was dissolved in H20 and the solution was acidified to pH 4 with dil HCl. EtOAc extraction, drying (MgSO4), filtration, and conce~ dLion gave the title compound ( 1.75 g, 57%) as a pale yellow solid: TLC Rf (4:20:20:56 MeOH/EtOAc/hexane/Cl2CH~) 0.22; 1~ NMR (CDCl3) o 1.55 (s, 9H), 1.56 (m, 2H), 1.80 (m, 2H), 3.01 (m, 4H), 3.55 (m, 2H), 3.70 (s, 3H), 25 4.05 (m, lH), 4.40 (m, lH), 4.55 (m, lH), 6.81 (d, J = 8.4 Hz, lH), 7.70 (s, lH), 7.89 (d, J = 8.4 Hz, lH).
b) Methyl (S)-7-(tert-butoxycarbonyl)-2,3,4,5-tetrahydro-3-oxo-4-(4-hydroxybut- 1 -yl)- 1 H- 1,4-benzodi~epine-2-acetate To a solution of N-[[2-(N-4-hydroxybut-1-yl)aminomethyl-4-tert-butoxycarbonyl]phenyl]-L-aspartic acid ,B-methyl ester (1.75 g, 4.1 mmol) and triethylamine (1.15 mL, 8.2 mmol) in dichloromethane (150 mL) under argon at RT
was added benzotriazol- 1 -yloxy-tris(dimethylamino)phosphonium hexafluorophosphz-~ (2.08 g, 14.7 mmol). The reaction mixture was stirred 35 overnight at RT, then was washed sequentially with ice-cold dil HCl, water, 5%
sodium bicarbonate, saturated brine, and then dried (MgS04). Filtration and concentration left a residue which was purified by silica gel flash column W O 97/24119 PCT~US96/20748 chromatography (5% mPth~nol:ethyl acetate) to give the title compound (0.631 g, 38%): TLC Rf (4% MeOH/EtOAc) 0.26; IH NMR (CDC13) o 1.46-1.61 (m, 4H), 1.57 (s, 9H), 2.64 (d, J = 6.9 Hz, lH), 2.66 (dd, J = 15.9, 6.3 Hz, lH), 2.99 (dd, J = 15.6, 6.9 Hz, lH), 3.56-3.54 (m, 4H), 3.74 (s, 3H), 3.84 (d, J = 16.2 Hz, lH), 4.54 (m, lH), 5.10 (m, lH), 5.41 (d, J = 16.2 Hz, lH), 6.49 (d, J = 8.3 Hz, lH), 7.59 (d, J = 1.8 Hz, lH), 7.67 (dd, J = 8.3, 1.8 Hz, lH); MS (ES) m/e 407.2 [M+H]+; ~CC~D
= - 185.4~ (c = 1, CH30H).
c) Methyl (S)-7-(tert-butoxycarbonyl)-2,3,4,5-tetrahydro-3-oxo-4-(4-phth~limic~obut-1-yl)-lH-1,4-benzodiazepine-2-acetate To a solution of methyl (S)-7-(tert-butoxycarbonyl)-2,3,4,5-tetrahydro-3-oxo-4-(4-hydroxybut-1-yl)-lH-1,4-benzodiazepine-2-acetate (437 mg, 1.07 mmol) and triphenylphosphine (308 mg, 1.17 mmol) in THF (20 mL) at RT under argon was sequentially added phth~limi~ (173 mg, 1.17 mmol) and diethyl azodicarboxylate (205 mg, 1.17 mmol). The reaction ll~ib~luie was stirred overnight at RT, the solvent was removed, and the residue was purified by silica gel flashcolumn chromatography (4:20:20:56 m~ths~n~ll/ethyl acetatethexane/methylene chloride) to give the tit}e compound (0.430 g, 75%): TLC Rf (4:20:20:56 MeOH/EtOAc/hexane/Cl2CH2) 0.32; IH NMR (CDCl3) o 1.55 (s, 9H), 1.55-1.61 (m, 4H), 2.68 (dd, J = 14.0, 5.7 Hz, 2H), 2.98 (dd, J = 14.0, 6.6 Hz, lH), 3.46-3.64 (m, 4H), 3.71 (s, 3H), 3.85 (d, J = 16.5 Hz, lH), 4.63 (d, J = 4.4 Hz, lH), 5.08 (dd, J =
5.7, 6.6 Hz, lH), 5.37 (d, J = 16.5 Hz, lH), 6.48 (d, J = 8.3 Hz, lH), 7.67 (s, lH), 7.69 (d, J = 8.3, 1.8 Hz, lH), 7.72-7.76 (m, 2H), 7.81-7.86 (m, 2H).
d) Methyl (S)-2,3,4,5-tetrahydro-7-carboxy-3-oxo-4-(4-phth:~limidobutyl)-lH-1,4-benzodiazepine-2-acetate To a solution of methyl (S)-7-(tert-butoxycarbonyl)-2,3,4,5-tetrahydro-3-oxo-4-(4-phth~limidobutyl)-lH-1,4-benzodiazepine-2-acetate (660 mg, 0.89 m~nol) in dichloromethane (20 mL) was added 4 N HCI/dioxane (5 mT, 20 mmol) at RT
under argon. The reaction mixture was stirred for 18 hr. The s~lspen~ion was concentrated to give the title compound as an off-white solid (425 mg, 98%): IH
NMR (CDCl3) ~ 1.55-1.61 (m, 4H), 2.71 (dd, J = 14.1, 6.0 Hz, 2H), 3.01 (dd, J =
14.1, 6.3 Hz, lH), 3.50-3.65 ~m, 4H), 3.75 (s, 3H), 3.89 (d, J = 16.5 Hz, lH), 4.68 - (d, J = 4.5 Hz, lH), 5.12 (dd,3 = 6.0, 6.3 Hz, lH), 5.40 (d, J = 16.6 Hz, lH), 6.41 - 35 (bs, lH), 6.53 (d, J = 8.4 Hz, lH), 7.69-7.75 (m, 4H), 7.82-7.85 (m, 2H); MS (l~S) m~e 480.2 [M+H]+.
W O 97/24119 PCT~US96/20748 e~ Methyl (S)-2,3,4,5-tetrahydro-7-[[[(b~n7imi-1~7Ol-2-yl)methyl3amino]carbonyl]-3-oxo~-(4-phth:~limiclobut-1-yl)-lH-1,4-benzodiazepine-2-acetate EDC (240 mg, 1.25 mmol) was added to a stirred solution of methyl (S)-2,3,4,5-tetrahydro-7-carboxy-3-oxo-4-(4-phth~limi~lQbut-l-yl)-lH-1,4-benzodiazepine-2-acetate (0.85 g, 0.88 mmol), 2-(aminomethyl)'oe~.7i.,~ 7Ole dihydrochloride (230 mg, 1.04 mmol), HOBt -H2O (169 mg, 1.25 mmol), and diisopropylethylamine (0.78 mL, 4.5 mmol) in anhydrous acetonitrile (10 rnL) at RT. After 19 h, the reaction was concentrated on the rotavap (high vacuum), and the residue was partitioned between H20 (5 mL) and EtOAc (20 mL). The layers were separated and the organic layer was washed with H2O (5 rnL). Drying (MgSO4), concentration, and silica gel chromatography (5% MeOH/CH2Cl2), gave the title compound (230 mg, 43%) as an off-white solid: TLC Rf (5% MeOH/Cl2CH2) 0.30;
lH NMR (CD30D) o 1.42-1.56 (m, SH), 2.63 (dd, J = 6.4, 16.2 Hz, lH), 2.95 (dd, J= 6.7, 16.2 Hz, lH), 3.33-3.40 (m, 2H), 3.48-3.55 (m, 2H), 3.57 (d, J = 16.5 Hz, lH), 3.67 (s, 3H), 4.72~.80 (m, 3H), 5.03 (dd, J = 6.4, 6.7 Hz, lH), 5.20 (d, J = 16.5 Hz, lH), 6.44 (d, J = 8.4 Hz, lH), 7.18-7.21 (m, 2H), 7.52-7.63 (m, 6H); 7.74-7.76 (m, 2H), 9.08 (br s, lH).
f) (S)-2,3,4,5-Tetranydro-7-[t[(b~n7imicl~7ol-2-yl)methyl]amino]carbonyl]-3-oxo-4-[4-[(2-carboxybenzoyl)amino]but-1-yl]- lH-1,4-bt-.n7oflizl7~pine-2-acetic acidLiOH (30 mg, 0.71 mmole) was added at RT to a solution of methyl (S)-2,3,4,5-tetrahydro-7-[[[(benzimidazol-2-yl)methyl]amino]carbonyl]-3-oxo-4-(4-phth:~limi~lobut-l-yl)-lH-1,4-benzodiazepine-2-acetate (223 mg, 0.33 mmol) in MeOH (2 mL) and H20(3 mL). The reaction mixture was sti;red at RT for 19 hr.
Acidification with dil HCl to pH 4 and concentration produced a solid. Filtration gave the title compound (145 mg, 66%) as a white solid: ~a]D - -100.4~ (c = 1, CH30H); IH NMR (CD30D) ~ 1.32-1.65 (m, 5EI), 2.58 (dd, J = 16.4, 6.7 Hz, lH), 2.90 (dd, J = 16.4, 7.9 Hz, lH), 3.06 (m, lH), 3.69 (m, lH), 4.00 (d, J = 16.7 Hz, lH),4.69(brs,2H),5.11 (dd,J=7.9,6.7Hz, lH~, 5.36(d,J= 16.7Hz, lH),6.50 (d, J = 8.4 Hz, lH), 7.29 (m, 2H), 7.37 (m, 4H); 7.51 (m, 2H), 7.664 (s, lH), 7.74 (d, J = 6.8 Hz, lH); MS (ES) m/e 613.2 [M+H]+. Anal. Calcd for C32H32N607 -1.5 H2O: C, 60.0g; H, 5.51; N, 113.14. Found: C, 59.77; H, 5.46; N, 12.98.
E~ ?le 17 W O 97/24119 PCTrUS96/207~8 F'lc~aldlion of (+)-7-r3-(benzill,idazol-2-yl)propyll-4-methyl-3-oxo-2.3.4.5-tetrahydro- 1 H- 1.4-benzodiazepine-2-acetic acid a) Methyl (_)- 1 -(tert-butoxycarbonyl)-7-(4-hydroxy- 1 -butyn- 1 -yl)-4-methyl-3-oxo-2,3,4,5-tetrahydro- 1,4-benzodiazepine-2-acetate 3-Butyn-l-ol (65 mg, 0.93 mmol), bis(triphenylphosphine)pS~ m (II~
chloride (5 mg, 0.007 mmol), triphenylphosphine (10 mg, 0.038 mmol), and copper(I) iodide (10 mg, 0.052 mmol) were added under an Ar atmosphere to a solution of methyl (+)-2,3,4,5 -tetrahydro- 1 -(tert-butoxycarbonyl)-7-iodo-4-methyl-3-oxo-lH-1,4-benzodiazepine-2-acetate (obtained as in example 7c; 440 mg, 0.94 mmol) in triethylamine (34 m~ ). The reaction mixture was heated to reflux for 4 hr, then was filtered through celite(~), and the filtrate was concentrated. The residue was purified by silica gel flash column chromatography (4:20:20:56 MeOH/EtOAc/hexane/CI2CH2) to give the title compound (390 mg, g4%) as a pale yellow liquid: TLC Rf (5% MeOH: Cl2CH2) 0.37; IH NMR (400 MHz, CDCl3) o 7.34 (dd, J = 1.8, 8.1 Hz, lH), 7.31 (d, J = 1.8 Hz, lH), 7.13 (d, J = 8.1 Hz, lH), 5.15-5.23 (m, lH), 4.75 (d, J = 14.4 Hz, lH), 3.68 (d, J = 14.4 Hz, lH), 3.63-3.67 (m, 2H), 3.59 (s, 3H), 3.04, (s, 3H), 2.88 (dd, J = 5.5, 15.2 Hz, lH), 2.45 (t, J = 6.4 Hz, 2H), 2.26 (dd, J = 9.5, 15.2 Hz, lH), 2.04 (br s, lH), 1.34 (br s, 9H); MS (ES) m/e 417 [M+H]+.
b) Methyl ( )- 1 -(tert-butoxycarbonyl)-7-(4-hydroxybut- 1 -yl)4-methyl-3-oxo-2,3,4,5-tetrahydro- 1,4-benzodiazepine-2-acetate 10% Pd/C (40 mg) was added to a solution of methyl (+)-l-(tert-butoxycarbonyl)-7-(4-(hydroxy- 1 -butyn- 1 -yl)-4-methyl-3-oxo-2,3,4,5-tetrahydro-1,4-benzodiazepine-2-acetate (370 mg, 0.89 mmol) in EtOH (20 rnL), and the mixture was shaken on a Parr apparatus at RT under H2 (50 psi). After 12 h, the reaction was filtered through celite~, and the filtrate was concentrated to afford the title compound (350 mg, 94%) as a pale yellow liquid: TLC Rf (4:20:20:56 MeOH/EtOAc/hexane/CI2CH2) 0.55; IH NMR (400 MHz, CDCl3) o 7.10-7.19 (m, 3H), 5.59-5.77 (m, lH), 4.85 (d, J = 15.0 Hz, lH), 3.68-3.60 (m, SH), 3.12 (s, 3H), 2.85 (dd, J = 5.4, 15.3 Hz, lH), 2.65 (t, J = 6.4 Hz, 2H), 2.34 (dd, J = 10.0, 15.3 Hz, lH), 1.30-1.78 (m, 13H); MS (ES) rn/e 421 LM+H]+.
-35 c) Methyl (_)-l-(tert-butoxycarbonyl)-7-(4-carboxybut-1-yl)-4-methyl-3-oxo-2,3,4,5-tetrahydro- 1,4-benzodiazepine-2-acetate ~33 W O 97/24119 PCTrUS96/20748 To a solution of methyl (+)-l-(tert-butoxycarbonyl)-7-(4-hydroxybut- I -yl)4-methyl-3-oxo-2,3,4,5-tetrahydro- l ,4-ben7orli~7epine-2-acetate (350 mg, 0.82 mmol) in CH2Cl2 at 0~C was added 2,2,6,6-tetrarnethyl-oxopiperi~linillm chloride (J. Org. C*em. 1985, 50, 3930-3931; 220 mg, 1.1 rnmol).
S The mixture was stirred for 2 hr at 0~C under Ar atmosphere. 2-Methyl-2-butene (1 mL) was added, followed by the addition of a freshly prepared solution of NaClO2~0.76 g, G.7 mmol), NaH2PO, ~ H20 (0.78 g, 5.68 mrnol) and H20 (25 mL). The cooling bath was removed, and the rnixture was taken up in EtOAc and washed successively with 0.05 M HCl and brine. Drying (MgSO4), concentration, and silica gel chromatography (5% AcOH in 4:20:20:56 MeOH/EtOAc/hexane/Cl2CH2), gave the title compound (350 mg, 98%): TLC Rf (5% AcOH in 4:20:20:56 MeOH/EtOAc/hexane/Cl~CH2) 0.32; IH NMR (400 MHz, CDC13) o 7.13-7.18 (m, 3H), 5.60-5.69 (m, lH), 4.83 (d, J = 14.2 Hz, lH), 3.76 (d, J = 14.2 Hz, lH), 3.66 (s, 3H), 3.12 (s, 3H), 2.93 (dd, J = 4.5, 15.3 Hz, lH), 2.67 (t, J = 7.3 Hz, 2H), 2.36 (t, J
= 7.3 Hz, 2H), 2.30-2.34 (m, lH), 1.95 (q, J = 7.3 Hz, 2H), 1.34 (s, 9H); MS (ES) m/e 435 [M+H]+.
d) Methyl (+)-l-(tert-butoxycarbonyl)-7-t3-(~ 7.ol-2-yl)propyl]-4-methyl-3-oxo-2,3,4,5-tetrahydro- 1,4-benzodiazepine-2-acetate To a stirred and cooled (-10~C) rnixture of methyl (+)-l-(tert-butoxycarbonyl)-7-(4-carboxybut- 1 -yl)-4-methyl-3-oxo-2,3,4,5-tetrahydro- 1,4-benzodiazepine-2-acetate (350 mg, 0.8 mmol) and Et3N (81 mg, 0.8 rnrnol) in anhydrous THF (8 mL) was added isobutylchloroformate (97 mg, 0.8 mrnol). After 10 min, a solution of 1,2-phenylenefli:~min~ (1.43 g, 0.9 mmol) in THF (2 rnL) was added. Stirring was continued at RT overnight, then the solvents were evaporated.
The residue was dissolved in EtOAc, and the solution was washed seqllentiz3lly with aqueous NaHCO3 and brine. Drying (MgSO4) and concentration produced a pale yellow solid. This was dissolved in glacial AcOH (5 rnL), and the reaction was heated to 60~C. After 3 hr, the mixture was cooled, concentrated, neutralized with 2.5 N NaOH, and extracted with CH2C12. Drying (MgSO4), concentration, and silica gel chromatography (gradient 1-5% MeOH/CH2C12) gave the title compound (200 mg, 50%): TLC Rf (4:20:20:56 MeOH/EtOAc/hexane/Cl2CH2) 0.18; lH NMR
(400 MHz, CDC13) o 7.57-7.61 (m, 2H), 7.15-7.28 (m, 4H), 7.08 (s, lH), 5.60-5.55(m, lH), 4.75-4.88 (m, lH), 3.71 (d, J = 14.2 Hz, lH), 3.70 (s, 3H), 3.10 (s, 3H), 2.94 (dd, J = 4.2, 15.3 Hz, lH), 2.85-2.90 (m, 2H), 2.73-2.79 (m, 2H), 2.32-2.36 (m, lH), 2.15-2.23 (m, lH), 1.34 and 1.55 (br s, rotamers, 9H); MS (ES) m/e 507 [M+H]+.
W O 97/24119 PCTrUS96/20748 e) (+)-7-[3-(Ben7imi(i~7ol-2-yl)propyl~-4-methyl-3-oxo-2~3~4~s-tetrahydro- lH- 1,4-benzodiazepine-2-acetic acid LiOH (17 mg, 0.71 mmole) was added at RT to a solution of methyl (+)- 1-(tert-butoxycarbonyl)-7-[3-(ben7imi~1~7.ol-2-yl)propyl]-4-methyl-3-oxo-2,3,4,5-tetrahydro-1,4-benzodiazepine-2-acetate (200 mg, 0.395 mmol) in MeOH (2 mL) and H20 (3 mL). The reaction mixture was stirred at RT for 4 hr. Aci~lifi~tion with dil HCl to pH 4 and concentration produced a white solid. This was dissolved in a mixture of methylene chloride (10 mL) and trifluoroacetic acid (5 mL) at 0~C, and the reaction was kept at 0~C for 30 min. The solvents were ~v~olat~d and theresidue was triturated with ether and purified by ODS flash chromatography (gradient 10 to 18% CH3CN/H2O cont~ining 0.1% TFA). Concentration and lyophilization gave the title compound (75 mg, 48%) as a colorless powder: IH
NMR ((400 MHz, CDC13) â 7.58-7.61 (m, 2H), 7.37-7.39 (m, 2H), 6.79 (d, J = 8.2 Hz, lH), 6.68 (s, lH), 6.39 (d, J = 8.2 Hz, lH), 5.18(d, J = 16.8 Hz, lH~, 4.77 (dd, J
= 7.0, 7.5 Hz, lH), 3.63 (d, J = 16.8 Hz, lH), 3.03-3.19 (m, 3H), 2.97 (s, 3H), 2.83 (dd, J = 7.0, 16.4 Hz, lH), 2.52-2.56 (m, 2H), 2.08-2.12 (m, 2H); MS (ES) m/e 393.0 tM+H]~- Anal. Calcd for C22H24N4O3 ~-C2HF3O2 ~ 0.5 H2O: C, 55.92; H, 5.08; N, 10.89. Found: C, 56.16; H, 4.92; N, 10.88.
Example 18 Plc~paldtion of (S)-2.3.4.5-tetrahydro-7-rrN-r(ben7imi~1~7-)1-2-yl)mPtllyl~-N-(4-aminobutyl)aminolcarbonyll-4-methyl-3-oxo- 1 H- 1.4-benzodiazepine-2-acetic acid a) 4-[(B~n7imid~7OI-2-yl)methylJaminobutyronitrile To a stirred mixture of 2-aminomethylben7imi~i~701e dihydrochloride (0.5 g, 2.2717 mmol) and NaHCO3 (0.67 g, 7.951 mmol) in dry DMF (10 mL) was added 4-bromobutyronitrile (0.37 g, 2.4989 mmol). After stirring at RT for 24 hr, the mixture was concentrated. The residue was taken up in H2O and extracted with CH2Cl2. The organic extracts were dried over MgS04, concentrated, and purified by silica gel flash column chromatography (5% MeOH/CH2Cl2)to give the title compound (0.15 g, 35%) as a brown oil: 'H NMR (250 MHz, DMSO-d6) ~ 1.82 (m, - 2H), 2.45 (t, J = 4 Hz, 2H~, 2.85 (t, J = 4 Hz, 2H), 4.11 (s, 2H), 7.14 (m, 2H), 7.50 35 (m, 2H).
-W O 97/24119 PCTrUS96/20748 b) Methyl (S)-2,3,4,5-tetrahydro-7-[[N-[(benzimidazol-2-yl)methyl]-N-(4-cyanopropyl)amino]carbonyl]-4-methyl-3-oxo- lH- 1,4-benzodiazepine-2-acetate To a stirred mixture of 4-~(b~n7im;~ 7Ol-2-yl)methyl]aminobutyronitrile (0.159 g, 0.7422 mmol), methyl 2,3,4,5-tetrahydro-7-carboxy-4-methyl-3-oxo-lH-1,4-benzodiazepine-2-acetate ( 0.217 g, 0.7422 mmol), HOBt H,O (0.120 g, 0.8906 mmol), and i-Pr2NEt (0.192 g, 1.4844 mmol) in dry CH~CN (7 mL) was added EDC
(0.265 g, 0.8906 mmol). After stirring at RT for 48 hr, the ~ lule was concentrated. The residue was taken up in H2O and extracted with CH2Cl2. The organic layer was washed se~uentially with saturated NaHCO3 and brine, dried over MgSO4, and concentrated to give a brown oil. Silica gel flash column chromatography (3% MeOH/CH2CI2) gave the title compound (0.261 g, 74%) as an off white foam: 'H NMR (250 MHz, DMSO-d6): o 1.95 (m, 2H), 2.66 (dd, J = 16.4, 3.5 Hz, lH), 2.78 (dd, J = 16.4, 3.5 Hz, lH), 2.85 (t, J = 8.7 Hz, 2H), 3.45 (t, J = 8.7 Hz, 2H), 3.60 (s, 3H), 3.80 (d, J = 16 Hz, lH), 4.52 (s, 2H), 4.84 (d, J = 2.9 Hz, 2H), 5.15 (m, lH), 5.48 ~d, J = 16 Hz, lH), 6.40 (d, J = 3.5 Hz, IH), 6.54 (d, J = 8.3 Hz, lH), 7.25 (m, 4H);,7.50 (m, lH), 7.62 (m, lH).
c) (S)-2,3,4,5-Tetrahydro-7-[[N-[(ben7imi~1~7.ol-2-yl)methyl~-N-(4-cyanopropyl)amino]carbonyl~-4-methyl-3-oxo- lH- 1,4-benzodiazepine-2-acetic acidTo a stirred solution of methyl (S)- 2t3,4,5-tetrahydro-7-[~N-[(benzimidazol-2-yl)methyl]-N-(4-cyanopropyl)amino]carbonyl]-4-methyl-3-oxo- lH- 1,4-benzodiazepine-2-acetate (0.261 g, 0.5478 mmol) in MeOH (5 mL) was added 2.5 N
NaOH (0.7 mL, 1.6433 mmol). After stirring at RT overnight, the mixture was concentrated. The residue was taken up in H2O, and the solution was acidified with 6 N HC1 to pH = 4. The white solid was filtered and dried to afford the title compound ~0.21 g, 81%): 'H NMR (250 MHz, DMSO-d6): ;~ 1.95 (m, 2H), 2.66 (dd, J = 16.4, 3.5 Hz, lH), 2.78 (dd, ~ = 16.4, 3.5 Hz, lH), 2.85 (t, J = 8.7 Hz, 2H), 3.45(t,J=8.7Hz,2H),3.80(d,J= 16Hz, lH),4.52(s,2H),4.84(d,J=2.9Hz, 2H), 5.15 (m, lH), 5.48 (d, J = 16 Hz, lH), 6.40 (d, J = 3.5 Hz, lH), 6.54 (d, J = 8.3 Hz, lH), 7.25 (m, 4H), 7.50 (m, lH), 7.62 (m, lH~.
d) (S)-2,3,4,5-Tetrahydro-7-[~N-L(benzimidazol-2-yl)methyl]-N-(4-aminobutyl)amino]carbonyl]-4-methyl-3-oxo- lH- 1,4-benzodiazepine-2-acetic acid A mixture of (S)-2,3,4,5-tetrahydro-7-[[N-[(ben~i...i-1~7- 1-2-yl)methyl]-N-(4-cyanopropyl)amino]carbonyll-4-methyl-3-oxo- lH- 1,4-benzodiazepine-2-acetic acid(0.200 g, 0.4325 mmol) and NH40H (1 mL, 30% solution) in MeOH (5 mL) was hydrogenated over Ra/Ni at RT for 24 hr. The catalyst was ~lltered off, and the .
CA 0224l633 1998-06-26 W O97/24119 PCT~US96/20748 filtrate was concentrated and purified by reverse phase chromatography (10% CH3CN/H10 contzlining 0.1% TFA) to give the title compound (0.100 g, 33%) as an off white solid: 'H NMR ( 400 MHz, DMSO-d6) ~ 1.45 (m, 2H), 1.72 (m, 2H), 2.54 (dd, J = 16.4, 3.5 Hz, lH), 2.70 (m, 2H), 2.75 (dd, J = 16.4, 3.5 Hz, lH), 2.95 (s, 3H), 3.65 (t, J = 8.7 Hz, 2H), 3.85 (d, J = 16 Hz, lH), 5.05 (s, 2H), 5.15 (m, lH), 5.48 (d, J = 16 Hz, lH), 6.65 (d, J = 8.3 Hz, lH), 7.20 (m, 2H), 7.61 (m, 2H), 7.75 (s, 2H), 7.85 (m, 2H); IR (KBr) 3425, 3000, 3100, 1728, 1675, 1630, 1625, 1613 cm~';MS (ES) m/e 479 (M+H). Anal. Calcd for C25H3"N604 ~ 2 CF3CO2H: C, 49.30; H, 4.56; N,11.89. Found: C, 49.22; H, 4;89; N, 11.84.
Fx:-ml?le 19 Preparation of ~S)-2.3.4.5-tetrahydro-7-rrN-r(benzimidazol-2-yl)methyl-N-(2-cyanomethyl)amino~carbonyll-4-methyl-3-oxo- lH- 1.4-benzodiazepine-2-acetic acid a) t(Ben7imi(1~7Ol-2-yl)methyl]~minn~retonitrile Following the procedure of Example in 18(a), except substituting bromoacetonitrile for the 4-bromobutyronitrile, the title compound was prepared as an off white solid (0.15 g, 35%): lH NMR (250 MHz, DMSO-d6): o 3.71 (s, 2H), 3.98 (s, 2H), 7.14 (m, 2H), 7.50 (m, 2H).
b) Methyl (S)-2,3,4,5-tetrahydro-7-[[N-[(ben7imicl~7(-1-2-yl)methyl-N-(2-cyanomethyl)amino]carbonyl]-4-methyl-3-oxo- 1 H- 1,4-benzodiazepine-2-acetate Following the procedure of Example 18(b), except substituting [(ben7imi~701-2-yl)methyl]~mino~retQnitrile for 4-l (bçn7imi~1~7ol-2-yl)methyl]aminobutyronitrile, the title compound was prepared as an of ~ white foam (0.487 g, 66%): 'H NMR (250 MHz, DMSO-d6) ~ 2.66 (dd, J--16.4, 3.5 Hz, lH), 2.78 (dd, J = 16.4, 3.5 Hz, lH), 2.92 (s, 2H), 3.60 (s, 3H), 3.80 (d, J = 16 Hz, lH), 4.52 (s, 2H), 4.84 (d, J = 2.9 Hz, 2H), 5.15 (m, lH), 5.48 (d, J = 16 Hz, lH), 6.40 (d, J = 3.5 Hz, lH), 6.54 (d, J = 8.3 Hz, lH), 7.25 (m, 4H), 7.50 (m, 2H), 7.62 (m, 2H).
c) (S)-2,3,4,5-Tetrahydro-7-[[N-[(b~n,; . . ,i~1~7ol-2-yl)methyl-N-(2-cyanomethyl)amino]cal L o~yl]-4-methyl-3-oxo- 1 H- 1,4-benzodiazepine-2-acetic acid Following the procedure in Example 18(c), methyl (S)-2,3,4,5-tetrahydro-7-~ 35 [[N-[(bell~i.. id~7ol-2-yl)methyl-N-(2-cyanomethyl)amino]carbonyl]-4-methyl-3-- oxo-lH-1,4-benzodiazepine-2-acetate was saponified, and the product was y~ ~;.lli7e~l from EtOH, to give the title compound as a white solid (0.420 g, W O 97/24119 PCTrUS96/20748 89%): 'H NMR ( 400 MHz, DMSO-d6) ~i 2.66 (dd, J = 16.4, 3.5 Hz, lH), 2.78 (dd, J = 16.4, 3.5 Hz, lH~, 2.92 (s, 2H), 3.80 (d, J = 16 Hz, lH), 4.52 (s, 2H), 4.84 (d, J = 2.9 Hz, 2H), 5.15 (m, lH), 5.48 (d, J = 16 Hz, lH), 6.40 (d, J = 3.5 Hz, lH), 6.54 (d, J = 8.3 Hz, lH), 7.25 (m, 4H), 7.50 (m, lH), 7.62 (m, lH); MS (ES)m/e 465 (M+H)~. Anal. Calcd for C23H22N6O4 ~ 2 HCl: C, 53.19; EI, 4.66; N, 16.18;
Found: C, 52.98; H, 4.43; N, 16.53.
Fx~ ?le 20 Preparation of (S)-2.3.4.5-tetrahydro-7-rr~ben~ ol-2-yl)methyl~methyiamino~carbonyll-3-oxo-lH-l~4-benzo~ n~-2-acetic açid a) Dimethyl (R)-2-trifluoromethanesulfonylsuccinate To a stirred, cooled (0~C) mixture of dimethyl D-malate (5.5 g, 33.9213 mmol), and dry pyridine (2.82 g, 35.7164 mmol) in dry CH2Cl2 (55 mL) was added trifluoro..,t~lh~neslllfionic anhydride (10.0 g, 35.7164 mmol) dropwise. After stirring at 0~C for 4 hr, the ~ ule was cluenched with H~O and the layers were separated.The organic layer was washed sequentially with dil HCI and brine, dried over MgSO4, and concentrated to give title compound (8.50 g, 96%) as a white solid: 'H
NMR( 250 MHz, CDCI3: ~ 3.10 (d, J = 5.8 Hz, 2H), 3.74 (s, 3H), 3.78 (s, 3H), 5.52 (t, J = 5.8 Hz, lH).
b) Dimethyl D-(2-cyanophenyl)malate A mixture of 2-arninobenzonitrile (0.5 g, 4.2323 mmol), 2,6-di-tert-bu~ylpylidine (0.85 g, 4.4439 mmol), and dimethyl (R)-2-trifluorometh~n~slllfonylsuccinate in 2:1 hexane/chloroform (25 mL) was stirred at RT for 76 hr. The mixture was concentrated, and the residue was taken up in H2O
and extracted with EtOAc. The organic extracts were washed sequentially with 10%HCI and brine, dried over MgSO4, concentrated, and purified by silica gel flash column chromatography (10% EtOAc/hexane) to give the title compound (0.886 g, 80%) as a yellow solid: 'H NMR (250 MHz, CDCl3) ~ 2.95 (d, J = 5.8 Hz, lH), 3.74(s, 3H), 3.78 (s, 3H), 4.60 (m, lH), 5.28 (d, J = 5.8 Hz, lH), 6.73 (d, J = 8.5 Hz, lH), 6.80 (t, J = 8.5 Hz, lH), 7.47 (m, 2H).
c) Methyl (S)-2,3,4,5-tetrahydro-3-oxo-lH-1,4-benzodiazepine-2-acetate A solution of dimethyl D-(2-cyanophenyl)malate (10.75 g, 41.0006 mmol) in MeOH (pre-saturated with NH3 (g) for 10 min, 100 mL) was hydrogenated over CA 0224l633 l998-06-26 W O 97/24119 PCTrUS96/20748 Ra/Ni at 55 psi for 48 hr. The catalyst was filtered off, and the filtrate was concentrated and purified by silica gel flash colurnn chromatography (40%
EtOAc/hexane) to give the title compound (5.03 g, 53%) as an off white solid: 'HNMR (250 MHz, CDCl3) o 2.65 (dd, J = 16.3, 7.6 Hz, lH), 2.99 (dd, 16.3, 5.9 Hz, lH), 3.74 (s, 3H), 3.95 (dd, J = 16, 6.9 Hz, lH), 4.79 (m, lH), 4.95 (dd, J = 16, 5.3 Hz, lH), 6.55 (t, J = 5.3 Hz, lH), 6.65 (d, J = 7.6 Hz, lH), 6.78 (d, J = 7.6 Hz, lH), 6.97(d,J=7.6Hz, lH),7.15(d,J=7.6Hz, lH).
d) Methyl (S)-2,3 ,4,5-tetrahydro-7-bromo-3-oxo- 1 H- 1 ,4-benzodiazepine-2-acetate A mixture o~ methyl (S)-2,3,4,5-tetrahydro-3-oxo-lH-1,4-benzodiazepine-2-acetate (5.03 g, 21.4746 mmol) and n-Bu4NBr3 (10.35 g, 21.4746 mmol) in CHC13 (100 mL) was stirred at RT for 3 hr, then the ~ lule was concentrated. The residue was taken up in H20, stirred, and filtered to afford the title compound (5.61 g, 83~o) as an off white solid: 'H NMR (250 MHz, CDCl3) o 2.74 (dd, J = 16.3, 7.6 Hz, lH), 3.05 (dd, J = 16.3, 5.9 Hz, lH), 3.75 (s, 3H), 4.05 (dd, J = 16, 6.9 Hz, lH), 4.73 (t, J
=5.9Hz, lH),4.86(dd,J= 16,5.3Hz, lH),6.68(d,J=7.6Hz, lH),6.75 (t,J=5.3 Hz, lH), 7.14 (s, lH), 7.25 (d, J = 7.6 Hz, lH).
e) Methyl (S)-2,3,4,5-tetrahydro-7-[[N-[(ben7imi~1~71 1-2-yl)methyl]-N-methylamino]carbonyl]-3-oxo- 1 H- 1 ,4-benzodiazepine-2-acetate A mixture of methyl (S)-2,3,4,5-tetrahydro-7-bromo-3-oxo-lH-1,4-benzodiazepine-2-acetate (1.5 g, 4.77905 mmol), 2-(methylaminomethyl)ben~imi-1~7Qle dihydrochloride (2.24 g, 9.5809 mrnol), triphenylphosphine (1.26 g, 4.7905 mmol), n-Bu3N ( 6.21 g, 33.5333 mmol), and (Ph3P)4Pd (1.10 g, 0.9581 mmol) in N-methyl 2-pyrrolidinone (20 rnL) was flushedwith argon and carbon monoxide for 10 min. The mixture was then heated at 100-105~C under a carbon monoxide balloon for 8 hr. The mixture was cooled and acidified with 6 N HCl to pH = 2. The solution was extracted with EtOAc, and theEtOAc layer was discarded. The aqueous layer was neutralized with 30% NaOH
and extracted with CH2Cl2. The organic extracts were dried over MgSO4, concentrated, and purifled by silica gel flash column chromatography (5%
MeOH/CH2Cl2) to give the title compound (1.62 g, 80%) as an off white solid: 'H
NMR (250 MHz, DMSO-d6): ~; 2.65 (dd, J = 16.3, 7.6 Hz, lH), 2.81 (dd, J = 16.3, 5.9 Hz, lH), 3.05 (s, 3H), 3.60(s, 3H), 3.75 (dd, J = 16.3, 6.9 Hz, lH), 4.78 (s, 2H), 4.95 (m, lH), 5.05 (dd, J = 16, 5.3 Hz, lH), 6.20 (d, J = 5.9 Hz, lH), 6.55 (d, J = 7.6 Hz, lH), 7.25 (m, 4H), 7.55 (m, 2H), 8.21 (t, J = 5.3 Hz, lH).
W O 97/24119 PCT~US96/20748 f) (S)-2,3,4,5-Tetrahydro-7-[[[berl7imi(1:l7Ol-2-yl)methyl]methylamino]ccubollyl]-3-oxo- 1 H- I ,4-ben70~ 7epine-2-acetic acid Following the procedure of Example 18(c), methyl (S)-2,3,4,5-tetrahydro-7-[~N-t(ben7.imi(1~7.ol-2-yl)methyl]-N-methylamino]carbonyl]-3-oxo-lH-1,4-benzodiazepine-2-acetate was saponified to afford the title compound (0.060 g, 57%) as an off white solid: 'H NMR ( 400 MHz, DMSO-d6) ~ 2.52 (dd, J = 16.3, 7.6Hz, lH), 2.84 (dd, J = 16.3, 5.9 Hz, lH), 3.20 (s, 3H), 3.75 (dd, J = 16.3, 6.9 Hz, lH), 4.95 (t, J = 5.9 Hz, lH), 5.05 (dd~ J = 16, 5.3 Hz, lH), 5.10 (s, 2H), 6.59 (d, J =
7.6 Hz, lH), 7.12 (s, lH), 7.20 (d, J - 7.6 Hz, lH), 7.48 (m, 2H), 7.69 (m, 2H), 7.90 (d, J = 5.3 Hz, lH); IR ( KBr) 3600-3100, 3100-2800, 1681, 1613, 1601, 1485, 1445, 1314, 830, 764, 742 cm l; MS (ES) m/e 422 (M+H)~. Anal. Calcd for C2,H21N5O4: C, 61.91; H, 5.20; N, 17.19. Found: C, 61.57; H, 5.32; N, 17.29.
Fx~ml?le 21 Preparation of (S~-2.3.4.5-tetrahydro-7-rrrrl-(2-hydroxyethyl)ben7imi~1~7ol-2-yllmetllyllaminolcarbonyll4-methyl-3-oxo- lH- 1.4-benzodi~epine-2-acetic açid a) Ethyl 2-[[(ben7imi-1~7O1-2-yl)methyl]amino]acetate A mixture of 2-aminomethyll~e~ 701e dihydrochloride hydrate (4.0 g, 18.1736 mmol), NaHCO3 (7.63 g~ 90.868 mmol), and ethyl 2-bromoacetate (4.55 g, 27.2603 mmol) in dry DMF (60 rnL) was stirred at RT for 24 hr, then was concentrated. The residue was taken up in H~O and extracted with CH2CI2. The organic extracts were dried over MgSO4, concentrated, and purified by silica gelflLash column chromatography (5% MeOH/CH2Cl2) to give the title compound (0.50 g, 12%) as a brown oil: 'H NMR (250 MHz, CDCI3) ~ 1.95 (s, 3H), 3.48 (s, 2H), 4.50 (m, 4H), 7.25 (m, 2H), 7.35 (m, lH), 7.73 (m, lH).
b) Methyl (S)-2,3,4,5-tetrahydro-7-[[~[1 -(2-acetyloxyethyl)ben7imi~1~7ol-2-yl]methyl]amino]~l,onyl]4-methyl-3-oxo-lH-1,4-benzodiazepine-2-acetate Following the procedure of Example 18(b), except substituting ethyl 2-[[(benzimidazol-2-yl)methyl]amino]acetate for the 4-[(ben7imid~7ol-2-yl)methyl]aminobutyronitrile, the title compound (0.251 g, 78.5%) was prepared as a white solid: ;H NMR (250 MHz, CDCl3) ~ 1.95 (s, 3H~, 2.66 (dd, J = 16.4,3.5 Hz, lH), 2.95 (s, 3H), 3.05 (dd, J = 16.4, 3.5 Hz, lH), 3.60 (d, J = 16 Hz, lH), 3.75 (s, 3H), 4.45(d, J = 5.9 Hz, 2H), 4.62 (s, 2H), 4.92 (t, J = 5.9 Hz, 2H), 5.10 ( m, lH), .~
W O 97/24119 PCTrUS96/20748 5.40(d,J= 16Hz, lH),6.49(d,J=8.3Hz, lH),7.32(m,3H),7.60(m, 2H), 7.71 (m, lH), 8.15 (t, J = 5.3 Hz, lH).
c) (S)-2.3,4,5-Tetrahydro-7-[[[[1-(2-hydroxyethyl)bçn7imi-1~7ol-2-yl]methyl]amino]carbonyl]-4-methyl-3-oxo- 1 H- 1,4-benzodiazepine-2-acetic acid To a stirred, partial suspension of methyl (S)-2.3,4,5-tetrahydro-7-[[[[1-(2-acetyloxyethyl)ben7imi~ 7ol-2-yl]methyl]amino]carbonyl]-4-methyl-3-oxo-lH-1,4-benzodiazepine-2-acetate (0.241 g, 0.475 mmol) in THF (5 mL) was added 1.0 N
LiOH (1.4 mL, 1.425 mmol). After stirring at RT overnight, the mixture was concentrated. The residue was taken up in H2O and acidified with AcOH to pH = 4.The off-white solid was filtered and lliluldt~d with acetone to afford the titlecompound (0.16 g, 75%) as a white solid: 'H NMR (400 MHz, DMSO-d6):o 2.54 (dd, J = 16.4, 3.5 Hz, lH), 2.95 (s, 3H), 3.05 (dd, J = 16.4, 3.5 Hz, lH), 3.60 (d, J =
16 Hz, lH), 4.45 (d, J = 5.9 Hz, 2H), 4.62 (s, 2H), 4.92 (t, J = 5.9 Hz, 2H), 5.10 (m, lH),5.40(d,J= 16Hz, lH),6.49(d,J=8.3Hz, lH),7.32(m,3H),7.60(m,2H), 7.71 (m, lH), 8.15 (t, J = 5.3 Hz, lH); MS (ES) m/e 452 (M+H)+. Anal. Calcd for C23H25N5O5 - 0.75 H2O: C, 59.71; H, 5.72; N, 15.14. Found: C, 59.65, H, 5.70; N,14.88.
Exarnple 22 Preparation of (+)-2.3~4.5-tetrahydro-7-rrN-(b~n7imicl~7ol-2-yl)methyl-N-rr4-(2-carboxybenzoyl)amino~butyllaminolcarbonyll-3-oxo-4-(2-phenylethyl)- 1 H- 1.4-benzot1i~7~-pine-2-acetic acid a) 4-~[(Benzimidazol-2-yl)methyllamino]butylphthz~limide A mixture of 2-aminomethylbenzimidazole dihydrochloride hydrate (22.10 g, 100.7269 mmol), NaHCO3 (42.40 g, 503.6347 mmol), and 4-bromobutylphth~limicle (34.10 g, 120.8723 mmol) in dry DMF (250 mL) was heated at 100-110~C for 6 hr, then was cooled and concenllatt;d. The residue was taken up in H2O and extracted with CH2Cl2. The organic extracts were dried over MgSO4, concentrated, and purified by silica gel flash column chromatography (5%
MeOH/CH2Cl2) to give the title compound (10.8 g, 31%) as a brown foam: IH NMR
(250 MHz, CDCl3) ~ 1.65 (m, 2H), 1.85 (m, 2H), 2.75 (t, J = 8.9 Hz, 2H), 3.78 (t, J =
8.9 Hz, 2H), 4.17 (s, 2H), 7.20 (m, 2H), 7.60 (m, 2H), 7.72 (m, 2H), 7.88 (m, 2H).
-W O 97/24119 PCT~US96/20748 b) Methyl ~+)-2,3,4,5-tetrahydro-7-[[N-(ben~i.l.if1z-7Ql-2-yl)methyl-N-[~4-(phth~limido)butyl]amino]carbonyl]-3-oxo-4-(2-phenylethyl)-lH-1,4-benzodiazepine-2-acetate To a stirred mixture of 4-[[(~çn7imi~l~7Ql-2-yl3methyl]amino]butylphth:~1imi~ (1.75 g, 5.0525 mmol), methyl (+)-2,3,4,5-tetrahydro-7-carboxy-3-oxo-4-(2-phenylethyl)- 1 H- 1,4-benzodiazepine-2-acetate (1.61 g, 4.2104 mmol), HOBt H2O (0.69 g, 5.0525 mmol), and i-PrlNEt (1.10 g, 8.4209 mmol) in dry CH3CN (30 mL) was added EDC (1.50 g, 5.0525 mmol). After stirring at RT for 24 hr, the mixture was concentrated. The residue was taken up in H2O and extracted with CH2Cl2. The organic extracts were washed sequentially with saturated NaHCO3 and brine, dried over MgSO4, concentrated, and purified by silica gel ~lash column chromatography (5% MeOH/CH2Cl2) to give the title compound (2.85 g, 95%) as a yellow foam: 'H NM~ (250 MHz, DMSO-d6) o 1.60 (m, 2H), 2.65 (m, 2H), 2.85 (dd, J = 16.4, 3.5 Hz, lH), 3.55 (m, 4H), 3.65 (s, 3H), 4.00 (d, J=
16.0 Hz, lH), 4.18 (q, J = 8.9 Hz, 2H), 4.75 (s, 2H), 5.15 (m, lH), 5.45 (d, J = 16.0 Hz, lH), 6.23 (d, J = 5.3 Hz, lH), 6.57 (d, J = 7.6 Hz, lH), 7.20 (m, 7H), 7.55 (m, 4H), 7.90 (m, 4H).
c) Methyl (+)-tetrahydro-7-[[N-[ber-~mi~1~7Ol-2-yl)methyl]-N-~4-aminobutyl)amino]carbonyl~-3-oxo~(2-phenylethyl)- 1 H- 1,4-benzodiazepine-2-acetate A mixture of methyl (+)-2~3~4~5-tetrahydro-7-[[N-(ben7imi~ 7Ql-2-yl)methyl-N-[[4-(phth~limido)butyl]amino]carbonyl]-3-oxo-4-(2-phenylethyl)-lH-1,4-benzodiazepine-2-acetate (0 50 g, 0.7015 mmol) and hydrazine (0.07 g, 2.1045mmol) in MeOH (5 mL) was refluxed for 6 hr, then was cooled and concentrated.
The residue was taken up in H2O, acidified to pH = 2 with 6N HCl, and filtered to remove a white solid. This solid was discarded. The aqueous filtrate was extracted with EtOAc, and the EtOAc layer was discarded. The aqueous layer was basified topH = 9 with Na2CO3 and extracted with CHCl3. The organic layer was dried over MgSO~ and concentrated to give the title compound (0.41 g, 89%) as an off white solid: 'H NMR (250 MHz, DMSO-d6) ~ 1.47 (m, 2H), 1.75 (m, 2H), 2.65 (m, SH), 2.85 (dd, J = 16.3, 5.9 Hz, lH~, 3.65 (s, 7H), 4.05 (d, J = 16.0 Hz, lH), 5.05 (s, 2H), 5.15 (m, lH), 5.45 (d, J = 16, lH), 6.65 (d, J = 7.6 Hz, lH), 7.25 (m, 6H), 7.41 (s, lH), 7.60 ~m, 2H), 7.85 (m, 2H).
WO 97124119 rCT/US96/20748 d) (+)-2,3,4,5-Tetrahydro-7-[[N-(ben~i...icl~7Ql-2-yl)methyl-N-[[4-(2-carboxybenzoyl)amino]butyl]amino]carbonyl]-3-oxo-4-(2-phenylethyl)- 1 H- 1,4-benzodiazepine-2-acetic acid To a stirred solution of methyl (+)-tetrahydro-7-[[N-[ben7imi-1~7Ol-2-yl)methyl]-N-(4-aminobutyl)amino]carbonyl]-3-oxo-4-(2-phenylethyl)- 1 H- 1,4-benzodiazepine-2-acetate (0.34 g, 0.47 mmol) in THF was added 1.0 N LiOH (1.2 mL). After stirring at RT overnight, the mixture was concentrated, and the residue was acidified with AcOH to pH = 4. The solid was filtered and lliLuldt~d with acetone/ether to give the title compound (0.130 g, 39%) as a white solid: 'H
NMR(400 MHz, DMSO-d6) ~ 1.47 (m, 2H), 1.75 (m, 2H), 2.54 (dd, J = 16.3, 3.5 Hz, lH), 2.65 (m, 2H), 2.85 (dd, J = 16.3, 5.9 Hz, lH~, 3.20 (m, 2H), 3.75 (m, 4H), 4.05 (d, J = 16.0 Hz, lH), 5.05 (s, 2H), 5.15 (m, lH), 5.45 (d, J = 16, lH), 6.65 (d, J = 7.6 Hz, lH), 7.25 (m, 6H), 7.41 (s, lH), 7.60 (m, 2H), 7.85 (m, 2H); IR (KBr) 3400, 3326, 3100-3000, 1721, 1637, 1626, 1616, 1607,1300, 750, 694 cm~'; MS (ES) m/e 717 (M+H)~. Anal. Calcd for C40H38N6O8 - 3 H2O: C, 63.56; H, 5.87; N, 11.12.
Found: C, 63.56; H, 5.83; N, 11.04.
Fx~ml?le 23 Preparation of (+)-2.3.4.5-tetrahydro-7-rrN-(ben7imid~7ol-2-yl)methyl~-N-rr4-(4-azido-2-hydroxybenzoyl)aminolcarbonyll -3-oxo-4-(2-phenylethyl)- 1 H- 1.4-bell7odiazepine-2-acetic acid a) Methyl (+)-2,3,4,5-tetrahydro-7-~[N-(be~7imi~1~7Ol-2-yl)methyl]-N-[[4-(4-azido-2-hydroxybenzoyl)amino~carbonyl]-3-oxo-4-(2-phenylethyl)- 1 H- 1,4-benzodiazepine-2-acetate To a mixture of methyl (+)-2~3~4~s-tetrahydro-7-[[N-(brn7imi~l~7ol-2-yl)methyl-N-[[4-(phth~limido)butyl]amino]carbonyl]-3-oxo-4-(2-phenylethyl)-lH-1,4-benzodiazepine-2-acetate (0.409 g, 0.702 mmol), 4-~.ido.c~licylic acid-N-hydroxysuccinimide ester (0.194 g, 0.702 mmol), and i-Pr2NEt (0.272 g, 2.106 mmol) in dry 2: 1 CH3CN/DMF (10 mL) was added EDC (0.25 g, 0.8424 mmol).
After stirring at RT for 24 hr, the mixture was concentrated. The residue was taken up in H2O, stirred, and ~lltered to afford, after drying, the title compound (0.204 g, 39%) as an off white solid: 'H NMR (250 MHz, DM~O-d6) ~ 1.40 (m, 2H), 1.75 (m, 2H), 2.65 (m, 3H), 2.75 (dd, J = 16.3, 5.9 Hz, lH), 3.20 (m, 2H), 3.51 (m, 4H), 3.65 (m, 3H), 3.95 (d, J = 16 Hz, lH), 4.79 (s, 2H), 5.12 (m, lH), 5.37 (d, J = 16 Hz, lH), W O 97/24119 PCTrUS96/20748 6.20 (s, lH), 6.55 (m, 3H), 7.20 (m, 8H), 7.55 (m, 3H), 7.~5 (d, J = 7.6 Hz, lH), 7.98 (s, lH), 8.75 (s, lH).
b) (+)-2,3,4,5-Tetrahydro-7-[[N-(be~ 7nl-2-yl)methyl3-N-[[4-(4-azido-2-hydroxybenzoyl)amino]carbonyl]-3-oxo4-(2-phenylethyl)- 1 H- 1,4-benzodiazepine-2-acetic acid Following the procedure of Example 22(d), methyl (+)-2,3,4,5-tetrahydro-7-[[N-(benzimidazol-2-yl)methyl]-N-[[4-(4-azido-2-hydroxybenzoyl~arninoJcarbonyl]-3-oxo-4-(2-phenylethyl)-lH-1,4-benzodiazepine-2-acetate was saponified to affordthe title compound (0.100 g, 50%) as a white solid: 'H NMR (400 MHz, DMSO-d6) ~ 1.40 (m, 2H), 1.75 (m, 2H), 2.65 (m, 3H), 2.75 (dd, J = 16.3, 5.9 Hz, lH), 3.20 (m, 2H), 3.51 (m, 4H), 3.95 (d, J = 16 Hz, IH), 4.79 (s, 2H), 5.12 (m, lH), 5.37 (d, J =
16 Hz, lH), 6.20 (s, lH), 6.55 (m, 3H), 7.20 (m, 8H), 7.55 (m, 3H), 7.85 (d, J = 7.6 Hz, lH), 7.98 (s, lH), 8.75 (s, lH); MS (ES) m/e 730 (M~H)+. Anal. Calcd for C39H39N9O6 ~ 2.~ ~20: C, 60.46; H, 5.72; N, 16.27. Found: C, 60.46; H, 5.43; N, 15.90.
F~ nple 24 rl~palalion of 2.3.4.5-tetrahydro-7-rrN-r(ben7imi(1~7ol-~-yl)methyl-N-rrr(+)-biotinoyllaminolbutyll~minolcarbonyll-3-oxo-4-(2-phenylethyl)-lH-1.4-benzo~ pine-(2RS)-acetic acid a) Methyl 2,3,4,5-tetrahydro-7-[~N-[(benzimidazol-2-yl)methyl-N-[[[(+)-biotinoyl]amino]butyl]amino~carbonyl~-3-oxo-4-(2-phenylethyl)- l H- 1,4-benzodiazepine-(2RS)-acetate To a mixture of methyl (+)-tetrahydro-7-[[N-[ben7imirl~7ol-2-yl)methyl]-N-(~aminobutyl)arnino~carbonyl]-3-oxo-4-(2-phenylethyl)- 1 H- 1,4-benzodiazepine-2-acetate (0.40 g, 0.6865 mmol), (+)-biotin (0.17 g, 0.6865 mmol), HOBt ~ H2O (0.11 g, 0.8239 mmol), and i-Pr2NEt (0.18 g, 1.3730 mmol) in 1:2 DMF/CH3CN (12 rnL) was added EDC (0.25 g, 0.8239 mmol). After stirring at RT for 24 hr, the mixturewas concentrated. The residue was taken up in H2O and extracted with CHCl3. The organic extracts were washed sequentially with saturated NaHCO3 and brine, driedover MgSO4, concentrated, and purified by silica gel flash column chromatography(10% MeOH/CH2CI2~ to give the title compound (0.24 g, 44%) as a yellow foam: lH
NMR (250 MHz, DMSO-d6) o 1.30 (m, 2H), 1.60 (m, 4H), 2.05 (t, J = 8.9 Hz, 2H);,2.60 (m, 3H), 2.68 (dd, J = 16.3, 5.9 Hz, lH), 3.10 (m, 4H), 3.45 (m, 2H), 3.60 CA 0224l633 l998-06-26 W O 97/24119 PCT~US96/20748 (m, 2H), 3.65 (s, 3H), 4.01 (d, J = 16 Hz, lH), 4.12 (t, J = 8.9 Hz, lH), 4.30 (t, J = 8.9 Hz, lH), 4.78 (s, 2H), 5.10 (m, lH), 5.45 (d, J = 16 Hz, lH), 6.20 (d, J = 5.3 Hz, lH), 6.40 (d, J = 8.9 Hz, 2H), 6.55 (d, J = 7.6 Hz, lH), 7.25 (m, 9H), 7.45 (m, lH), 7.55 (m, lH), 7.70 (t, J = 8.6 Hz, lH).
b) 2,3,4,5-Tetrahydro-7-[[N-[(bçn7imicl~7ol-2-yl)methyl-N-r[[(+)-biotinoyl]amino]butyl]amino]carbonyl]-3-oxo-4-(2-phenylethyl)-lH-1,4-benzodiazepine-(2RS)-acetic acid To a stirred solution of methyl 2,3,4,5-tetrahydro-7-[[N-[(benzimidazol-2-yl)methyl-N-[[[(+)-biotinoyl]amino]butyl]amino]carbonyl]-3-oxo-4-(2-phenylethyl)-lH-1,4-benzodiazepine-(2RS)-acetate (0.24 g, 0.2967 mmol) in 1:2 THF/MeOH (6 mL) was added 1.0 N LiOH (0.44 mL). After stirring at RT overnight, the mixture was concentrated. The residue was taken up in H2O and acidified with AcOH to pH
= 4. The off-white solid was filtered, and triturated with hot acetone to give the title compound (0.160 g, 68%) as a white solid: 'H NMR (400 MHz, DMSO-d6) d 1.30 (m, 2H), 1.60 (m, 4H), 2.05 (t, J = 8.9 Hz, 2H), 2.60 (m, 3H), 2.68 (dd, J = 16.3, 5.9 Hz, lH), 3.10 (m, 4H), 3.45 (m, 2H), 3.60 (m, 2H), 4.01 (d, J = 16 Hz, lH), 4.12 (t, J
= 8.9 Hz, lH), 4.30 (t, J = 8.9 Hz, lH), 4.78 (s, 2H), 5.10 (m, lH), 5.45 (d, J = 16 Hz, lH), 6.20 (d, J = 5.3 Hz, lH), 6.40 (d, J = 8.9 Hz, 2H), 6.55 (d, J = 7.6 Hz, lH), 7.25 (m, 9H), 7.45 (m, lH), 7.55 (m, lH), 7.70 (t, J--8.6 Hz, lH); MS (l~S) m/e 795 (M+H)~. Anal. Calcd for C42H5"N806S 1.75 H20: C, 61.04; H, 6.52; N, 13.56.
Found: C, 60.89; H, 6.24; N, 13.31.
F.x~ml?le 25 P.epa~a~ion of (+)-2.3~4~5-tetrahydro-7-rrN-r(benzimidazol-2-yl)methyl~-N-(4-aminobutyl)~minolcarbonyll-3-oxo-4-(2-phenylethyl)-lH-1.4-benzodiazepine-2-acetic acid a) (+)-2,3,4,5-Tetrahydro-7-[[N-[(ben7imid~7ol-2-yl)methyl]-N-(4-aminobutyl)amino]carbonyl]-3-oxo-4-(2-phenylethyl)- lH- 1,4-benzodiazepine-2-acetic acid Following the procedure in Example 24(b), methyl (+)-2,3,4,5-tetrahydro-7-[[N-[ben7imifl~701-2-yl)methyl]-N-(4-aminobutyl)amino]carbonyl]-3-oxo-4-(2-phenylethyl)-lH-1,4-benzodiazepine-2-acetate was saponi~led to give the title compound (0.250 g, 80%) as an off white solid: 'H NM~ (400 MHz, DMSO-d6) 1.37 (m, 2H), 1.62 (m, 2H), 2.52 (dd, J = 3.5 Hz, lH), 2.64 (m, 2H), 2.75 (dd, J =
CA 0224l633 l998-06-26 W O 97/24119 PCT~US96/Z0748 16.3, 5.9 Hz, lH), 3.51 (m, 4H), 3.91 (d, J = 16 Hz, lH), 4.98 (s, 2H), 5.05 (m, lH),5.37(d,J= 16Hz, lH),6.53(d,J=7.9Hz,2H),7.17(m,7H),7.52(m, lH), 7.62 (s, lH), 7.78 (m, lH); IR ( KBr): 3386, 3100-3000, 1647, 1613, 1403, 740, 699 cm '; MS (ES) m/e 569 (M+H)+. Anal. Calcd for C32H36N604 ~ 2.75 H20: C, 62.18; H, 6.77; N, 13.60. Found: C, 62.11; H, 6.68; N, 13.57.
Fxzlrnple 26 Preparation of (+)-2~3.4~5-tetrahydrb-7-rfN-r(benzimidazol-2-yl)methyll-N-rr4-(4-azido-3-iodo-2-hydroxyben7oyl~aminolbutyllaminolc~1Jon~111-3-oxo-4-(2-ph~r~ylethyl)- 1 H- 1.4-ben~otli~7~pine-2-acetic acid a) 3-Iodo-4-~7i~1os~1icylic acid-N-hydroxysllccinimifl. ester To a stirred mixture of 4-azidosalicylic acid N-hydroxysuccinimide ester (0.500 g, 1.8103 mmol) and silver trifluoroacetate (0.44 g, 1.9913 mmol) in CHCI3 (10 mL) was added iodine (0.510 g, 1.9913 mmol). After stirring at RT overnight,the reaction was filtered to remove a solid precipitate. The filtrate was washedsequentially with H2O, saturated NaHCO3 and brine, then was dried over MgSO~.
Concentration gave the title compound (0.703 g, 97%) as a light purple solid: 'HNMR (250 MHz, CDCl3) ~ 2.98 (s, 4H), 6.83 (d, J = 7.6 Hz, lH), 8.05 (d, J = 7.6 Hz, lH).
b) Methyl (+)-tetrahydro-7-[[N-[(ben7imic1~7O1-2-yl)methyl]-N-[[4-(4-azido-3-iodo-2-hydroxybenzoyl)amino]butyl]arnino]carbonyl]-3-oxo-4-(2-phenylethyl)-lH-1.4-benzodiazepine-2-acetate Following the procedure in Example 22(b), except substituting 3-iodo-4-azidosalicylic acid-N-hydroxysuccinimide ester for the 4-[[(~n7imi~1~7ol-2-yl)methyl]amino~butylph~h~limicl~, the title compound (0.312 g, 56%) was prepared as a yellow foam: 'H NMR (250 MHz, DMSO-d6) ~ 1.42 (m, 2H), 1.60 (m, 2H), 2.52 (dd, J = 16.3, 3.5 Hz, lH), 2.63 (m, 2H), 2.79 (dd, J = 16.3, 5.9 Hz, lH), 3.25 (s, 2H), 3.55 (m, 6H), 3.65 (s, 3H), 3.95 ~d, J = 16 Hz, lH), 4.75 (s, 2H), 5.02 (m, lH), 5.35 (d, J = 16 Hz, lH), 6.14 (d, J = 5.3 Hz, lH), 6.52 (d, J = 7.9 Hz, lH), 6.86 (d, J = 7.9 Hz, lH~, 7.25 (m, lOH), 7.51 (s, 2H), 7.90 (d, J = 7.9 Hz, lH), 9.01 (s, lH).
W O 97/24119 PCTrUS96/20748 c) (+)-Tetrahydro-7-[[N-[(bçl ~ i . I ,i(1~7Ol-2-yl)methyl~-N-[~4-(4-a_ido-3-iodo-2-hydroxybenzoyl)amino]butyl] amino]carbonyl] -3 -oxo-4-(2-phenylethyl)- 1 H-1.4-ben7.o~ 7~pine-2-acetic acid Following the procedure in Example 22(c), methyl (+)-tetrahydro-7-[[N-[(bç~ 7Ol-2-yl)methyl]-N-[[4-(4-a7ido-3-iodo-2-hydroxybenzoyl)amino]butyl]amino]carbonyl] -3-oxo4-(2-phenylethyl)- 1 H- 1.4-- benzodiazepine-2-acetate was saponified. Purification by silica gel flash colurnn chromatography (0.5, 0.5, 9.5 AcOH/MeOH/CH2Cl2) gave the title compound (0.170 g, 58%) as an off white solid: 'H NMR (400 MHz, DMSO-d6) ~ 1.42 (m, 2H), 1.60 ~m, 2H), 2.52 (dd, J = 16.3, 3.5 Hz, lH), 2.63 (m, 2H), 2.79 (dd, J = 16.3, 5.9 Hz, lH), 3.25 (s, 2H), 3.55 (m, 6H), 3.95 (d, 16, lH), 4.75 (s, 2H), 5.02 (m, lH), 5.35 (d, J= 16Hz, lH),6.14(d,J=5.3Hz, lH),6.52(d,J=7.9Hz, lH),6.86(d,J=7.9 Hz, lH), 7.25 (m, 10H), 7.51 (s, 2H), 7.90 (d, J = 7.9 Hz, lH), 9.01 (s, lH); MS (ES) mle 856 (M+H)+; IR (KBr): 3360, 3100-3000, 2116, 1704, 1643, 1610, 1586, 1477, 1305, 1274, 766, 700 cm~'. Anal. Calcd for C39H38IN906 ~ 4.5 H20: C, 50.01; H, 5.06;
N, 13.46. Found: C, 50.19; H, 5.01; N, 13.12.
Fx~ml?le 27 Preparation of 5-rrr(ben7imid~7ol-2-yl)methyllmethylaminolcarbonyll-lH-be~ 7Ole-2-aminoacetic acid a) Methyl 5-[~[(benzimida_ol-2-yl)methyl]methylamino]carbonyl]-lH-benzimidazole-2-;~mino~cetate Diisopropylethylamine (1.1 mL, 6.48 mmol) was added to a stirred solution of methyl 5-carboxy-b~n7imi~1~7Ole-2-~mino~et~t~ (0.24 g, 0.96 mmol), 2-(methylaminomethyl)bellzil,.idazole bis-trifluoroacetate (0.56 g, 1.44 mmol), HOBt H2O (0.19 g, 1.44 mmol), and EDC (0.28 g, 1.44 mmol) in anhydrous DMF (8 mL) at RT. After 23 h, the reaction mixture was diluted with CH2Cll (100 mL) and washed sequentially with 5% NaHCO3 (30 mL) and brine (30 mL). Drying (MgSO4), concentration, and silica gel chromatography (10% MeOH/CH2Cl2) gave the title compound (0.16 g, 42%~ as an off-white solid: MS (ES) m/e 393.0 (M+H)+.
A
35 b) 5-[[[(Ben7imicl~701-2-yl)methyl]methylatnino~carbonyl]-lH-bçn7imic~7Ole-2- aminoacetic acid W O97/24119 PCT~US96~0748 1.0 N LiOH (1.0 rnL, 1.0 mmol) was added dropwise at RT to a mixture of methyl 5-[[r(ben7imid~7-~1-2-yl)methyl]methylamino]carbonyl]-lH-ben7.imi~1~701e-2-aminoacetate (0.16 g, 0.41 mmol) in THF (10 rnL) and H20 (10 mL). After 1 h, the reaction ~ Lule was concentrated to a small volume on the 5 rotavap and cooled in an ice bath before neutralizing with 1.0 N AcOH (1.0 mL).
The solid was collected, washed with cold H20, and air dried to give the title compound (0.15 g, 100%) as an off white solid: MS (ES) rn/e 379.2 (M+H)+.
~F~m,J?le 28 I~e~aldlion of (_)-2.3~4~s-Tetrakydro 7-rrrfbenzimidazol-2-yl)methyllmethy!;lmino~carbonyll-4-(3.3-~lim~thylbutyl)-3-oxo-lH-1.4-ben7n~ 7epine-2-acetic acid a) Methyl (V-2,3,4,5-tetrahydro 7-[[[(bc~7imi-1~7ol-2-yl)methyl]methylarnino]carbonyl]-4-(3,3-dimethylbutyl)-3-oxo- lH- 1,4-benzodiazepine-2-acetate Diisopropylethylamine (0.94 rnL, 5.4 mmol) was added to a stirred solution of methyl (_)-7-carboxy-4-(3,3 -dimethylbutyl)-3-oxo-2,3,4,5-tetrahydro- 1 H- l ,4-benzodia_epine-2-acetic acid (0.39 g, 1.08 mmol), 2-(methylamin~lllet}lyl)ben7imi~1~701e bis(trifluoroacetate) (0.42 g, 1.08 rnmol), ), EIOBt H2O (0.22 g, 1.62 mmol), and EDC (0.31 g, 1.62 mmol) in anhydrous DMF
(8 mL) at RT. After 23 h, the reaction mlixture was diluted with CH2Cl~ (100 mL)and washed sequentially with 5% NaHCO3 (2 x 25 ml) and brine (25 mL). Drying (MgSO4), concentration, and silica gel chromatography (7% MeOH/CH2Cl2) gave the title compound (0.39 g, 71%) as a white solid: MS (ES) m/e 506.4 (M+H)+.
b) (i)-2,3,4,5-Tetrahydro 7-~[[(bç~ 7Ol-2-yl)methyl]methylamino]carbonyl]-4-(3,3-dimethylbutyl) -3 -oxo- 1 H- 1,4-benzodia_epine-2-acetic acid 1.0 N LiOH (1.Q rnL, 1.0 mrnol) was added dropwise at RT to a mixture of methyl (i)-2,3,4,5-tetrahydro 7-~[(b~imi~7(~l-2-yI)methyl]methylamino]carbonyl]-4-(3,3-dimethylbutyl)-3-oxo- lH- 1,4-benzodiazepine-2-acetate (0.38 g, 0.75 mmol) in THF (10 mL) and H2O (10 mL).
After 50 min, the reaction mixture was concentrated to a small volume on the rotavap and cooled in an ice bath before neutralizing with 1.0 N AcOH (2.5 mL).
The solid was collected, washed with a cold H20, and air dried to give the titlecompound (0.27 g, 73%) as a white solid: MS (ES) rn/e 492.2 (M~H)+.
W O 97124119 PCTrUS96/20748 F.xample 29 Preparation (_)-2.3.4.5-Tetrahydro 7-rrr(ben7imi{~7ol-2-yl)methyl7aminolcarbonyll-4-(3 .3-dimethylbutyl)-3-oxo- 1 H- 1 .4-benzodiazepine-2-acetic acid -a) Methyl (+)-2,3,4,5-tetrahydro 7-[l[~be.n7imi~1~7~.1-2-yl)methyl]amino]carbonyl]-4-(3,3-dimethylbutyl)-3-oxo- lH-1,4-benzodiazepine-2-acetate Diisopropylethylamine (0.79 rnL, 4.56 mmol) was added to a stirred solution of methyl (+)-7-carboxy-4-(3 ,3-dimethylbutyl)-3-oxo-2,3,4,5-tetrahydro- lH- 1,4-benzodiazepine-2-acetic acid (0.33 g, 0.91 mmol), 2-(aminomethyl)ben7imi~1~7Ole dihydrochloride hydrate (0.3 g, 1.36 mmol), HOBt H2O (0.18 g, 1.36 mmol), and ED(~ (0.26 g, 1.36 mmol) in anhydrous DMF (8 mL) at RT. After 20 h, the reaction15 rnixture was diluted with CH Cl2 (70 mL) and washed sequentially with 5% NaHCO3 (2 x 20 mL) and brine (20 mL). Drying (MgSO4), concentration, and silica gel chromatography (7% MeOH/CH2Cl2) gave the title compound (0.25 g, 56%) as a white solid: MS (ES) m/e 492.4 (M+H)+.
20 b) (_)-2,3,4,5-Tetrahydro 7-[[[(be~ 7Ol-2-yl)methyllamino]carbonyl]4-(3,3-dimethylbutyl)-3-oxo-lH-1,4-benzotli:l7epin~-2-acetic acid 1.0 N LiOH (1.0 mL, 1.0 mmol) was added dropwise at RT to a mixture of methyl ( )-2,3,4,5-tetrahydro 7-[[[(benzimidazol-2-yl)methyl~amino]carbonyl]-4-(3,3-dimethylbutyl)-3-oxo-lH-1,4-benzodiazepine-2-acetate (0.24 g, 0.49 mmol) in25 THF (8 mL) and H2O (8 mL). After 2.5 h, the reaction mixture was concentrated to a small volume on the rotavap and cooled in an ice bath before neutralizing with 1.0 N AcOH (1.2 mL). The solid was collected, washed with cold H20 ,and air dried togive the title compound (0.25 g, 109%) as a white solid: MS (ES) m/e 478.2 (M+H)+.
l~xample 30 Plc;palation of (+)-2.3.4~5-Tetrahydro 7-rrr(4-azaben7.i~ 7ol-2-yl)methyllmethylaminolcarbonyl~-4-(3 .3-dimethylbutyl)-3-oxo- 1 H- 1.4-35 benzodiazepine-2-acetic acid CA 0224l633 l998-06-26 W O 97t24119 PCTrUS96/20748 a) Methyl (+)-2,3,4,5-tetrahydro 7-[[[(4-~7~hen7imi~l~7~ 2-yl)methyl]methylamino]carbonyl]-4-(3,3-dimethylbutyl)-3-oxo- 1 H- 1,4-benzodiazepine-2-acetate Diisopropylethylamine (0.53 mL, 3.0 mmol) was added to a stirred solution of methyl (+)-7-carboxy-4-(3,3-dimethylbutyl)-3-oxo-2,3,4,5-tetrahydro- IH- 1,4-benzodiazepine-2-acetic acid (0.22 g, 0.61 mmol), 2-(methylamino)methyl-4-azaben7imi~1~701e diacetate (0.29 g, 1.0 mmol), ~IOBt ~ H2O (0.12 g, 0.91 mmol),and EDC (0.17 g, 0.91 mmol) in anhydrous CH3CN (12 mL) at RT. After 21 h, the reaction mixture was conce~ dl~d~ diluted with CEI~Cl2 (100 mL), and washed sequentially with 5% NaHCO3 (2 x 20 mL) and brine (20 mL). Drying (MgSO4), concentration, and silica gel chromatography (7% MeOH/CH2Cl2) gave the title compound (0.147 g, 48%) as a white solid: MS (ES) m/e 507.4 (M+H)+.
b) (+)-2,3,4,5-Tetrahydro 7-[[[(4-~7~h~ 7ol-2-yl)methyl]methylamino]carbonyl]-4-(3,3-dimethylbutyl)-3-oxo-lH-1,4-benzodiazepine-2-acetic acid 1.0 N LiOH (0.69 mL, 0.69 mmol) was added dropwise at RT to a rnixture of methyl (+)-2,3,4,5-tetrahydro 7-[[[(4-azaben7i,..il1~7Ol-2-yl)methyl]methylamino]carbonyl]-4-~3,3-dimethylbutyl)-3-oxo-lH- 1,4-benzodiazepine-2-acetate (0.14 g, 0.276 mmol) in THF (8 mL) and H2O (8 mL).
After 2 h, the reaction n~ib~ ; was concentrated to a small volume on the rotavap and cooled in an ice bath before neutralizing with 1.0 N AcOH (0.69 mL). The solid was collected, washed with cold H20, and air dried to give the title compound (0.074 g, 54%) as a white solid: MS (ES) m/e 493.2 (M+H)+.
F;.x~ml?le 31 Preparation of (Sl-2~3.4.5-tetrahydro-7-rrrl l-r(ben7imicl~7Ql-2-yl~m~tl~yllben7imicl~7ol-2-yllmetllyllarnino~cal1,o,l~1l-4-methyl-3-oxo-lH-l ~4-benzodiazepine-2-acetic acid - a) Methy} (S)-2,3,4,5-tetrahydro-7-[[[[l-[(benzimid~ol-2-yl)methyl]bçn7imi~ ol-2-yl]methyl]amino~carbonyl]-4-methyl-3-oxo-lH-1,4-ben7O~ 7epine-2-acetate Diisopropylethylamine (0.27 mL, 1.53 mmol) was added to a stirred solution of methyl (S)-7-carboxy-4-methyl-3-oxo-2,3,4,5-tetrahydro- 1 H- 1,4-benzodi~epine-2-acetate trifluoroacetate (0.14 g, 0.34 mmol), 2-[[l-[(ben7imi(i~7Ol-2-yl)methyl]be~7imitl~7c~e]methyl~amine bis(trifluoroacetate) (0.17 g, 0.34 mmol), 150 W O 97/24119 PCTrUS96/20748 HOBt H2O (0.064 g, 0.48 mmol), and ~DC (0.091 g, 0.48 mmol) in anhydrous DMF (10 mL) at RT. After 22 h, the reaction mixture was concentrated, diluted with CH2Cl2 (70 mL), and washed sequentially with 5% NaHCO3 (2x30 mL) and brine (20 mL). Drying (MgSO4), concentration, and silica gel chromatography (7% MeOH/CH2Cl2) gave the title compound (0.080 g, 43%) as a white solid: MS
~ (ES) m/e 552.2 (M+H)+.
b) (S)-2,3,4,5-Tetrahydro-7-[[~rl-[(ben7imid~7-~1-2-yl)methyl]ben7imi~ 7ol-2-yl]methyl]amino]carbonyl]-4-methyl-3-oxo-lH-1,4-benzodiazepine-2-acetic acid 1.0 N LiOH (0.36 mL, 0.36 mmol) was added dropwise at RT to a rnixture of methyl (S)-2,3,4,5-tetrahydro-7-[[[[l-[(ben7i~ 7ol-2-yl)methyl]ben7imi~t~7Ql-2-yl]methyl]amino]carbonyl]-4-methyl-3-oxo- lH- 1,4-benzodiazepine-2-acetate (0.08g, 0.14 mmol) in THF (5 mL) and H2O (4 mL). After 2 h, the reaction mixture was concentrated to a small volume on the rotavap and cooled in an ice bath before neutralizing with 1.0 N AcOH (0.69 mL). The solution was lyophilized to a crude product (0.086 g) as a white powder. Purification on C-18 Bond Elute (0%-20%
CH3CN/H2O cont:~ining 0.1 % TFA) afforded the title compound as a white powder:
MS (ES) m/e 538.2 (M+H)+.
Example 32 Prep~r~tion of (S)-2.3.4.5-Tetrahydro-7-rrbisr(ben7imicl~7ol-2-yl)methyllarninolcarbonyll-4-methyl-3-oxo-lH-1.4-benzodiazepine-2-acetic acid a) Methyl (S)-2,3,4,5-tetrahydro-7-[rbis[(ben7imi~1~7ol-2-yl)methyl]amino~carbonyl]-4-methyl-3-oxo- 1 H- 1,4-benzodiazepine-2-acetate Diisopropylethylamine (0.22 mL, 1.29 mmol) was added to a stirred solution of methyl (S)-7-carboxy-4-methyl-3-oxo-2,3,4,5-tetrahydro- 1 H- 1,4-benzodiazepine-2-acetate trifluoroz-~-et~te (0.075 g, 0.26 mmol), bis[(ben7imi<1~7Ol-2-yl)methyl]amine tris(trifluoro~cet~te) (0.16 g, 0.26 mmol), HOBt ~ H2O (0.05 g, 0.36 mmol), and EDC (0.069 g, 0.36 mmol) in anhydrous CH3CN (10 mL) at RT. After 17 h, the reaction rnixture was concentldted, diluted with CH2Cl2 (80 mL), and washed sequentially with 5% NaHCO3 (2 x 20 mL) and brine (20 mL). Drying - (MgSO4), concentration, and silica gel chromatography (7% MeOH/CH2Cl2) gave the title compound (0.05 g, 36%) as a white solid: MS (ES) m/e 552.2 (M+H~+.
-:
W O 97/24119 PCT~US96/20748 b) (S)-2,3,4,5-Tetrahydro-7-[[bist(ben7irni~ 7nl-2-yl)methyl]amino]carbonyl]-4-methyl-3-oxo-lH-1,4-benzodia_epine-2-acetic acid 1.0 N LiOH (0.23 mL, 0.23 mmol) was added dropwise at RT to a mixture of methyl ~S)-2,3,4,5-tetrahydro-7-[[bis[(ben7imi~7ol-2-yl)methyl]amino]carbonyl]-4-methyl-3-oxo-lH-1,4-benzodiazepine-2-acetate (0.05 g, 0.09 mmol) in THF (6 mL) and H20 (4 rnL). After 1 h, the reaction ~l~ixAluie was concentrated to a small volume on the rotavap and cooled in an ice bath before neutralizing with 1.0 N
AcOH (0.3 mT.). The solid was collected, washed with cold H~O, and air dried to give the title compound (0.048 g, 98%) as a white solid: MS (ES) m/e 538.2 (M+H)+.
Fx~mple 33 P~pdld~ion of f_)-2.3.4.5-tetrahydro-7-rrbisr~ben7imi~7ol-2-yl)methyllamino~carbonyll-3-oxo-4-(2-phenethyl)-lH-1 ~4-benzofli~7~ 1~ine-2-acetic B~
a) Methyl (+)-2,3,4,5-tetrahydro-7-[[bis[(ben7imi<1~7Ol-2-yl)methyl]amino]carbonyl~-3-oxo-4-(2-phenethyl)-lH-1,4-berl7o~ 7Ppine-2-acetate Diisopropylethylamine (0.3 mT, 1.74 mmol) was added to a stirred solution of methyl (+)-7-carboxy-3-oxo-2-(2-phenylethyl)-2,3,4,5-tetrahydro- lH- 1,4-benzodiazepine-2-acetate (0.11 g, 0.29 rnmol), bis[(ben7imi-1~7Ol-2-yl)methyl]amine tris(trifluoroacetate) (0.18 g, 0.29 mmol), HOBt ~ H2O (0.058 g, 0.43 mmol), andEDC (0.083 g, 0.43 mmol) in anhydrous CH3CN (12 mL) at RT. After 21 h, the reaction mixture was concelltld~d, diluted with CH2Cl2 (100 rnL), and washed sequentially with 5% NaHCO3 (2 x 20 mL) and brine (20 mL). Drying (MgSO4), concentration, and silica gel chromatography (7% MeOH/CH2CI2) gave the title compound (0.13 g, 70%) as a white solid: MS (ES) m~e 642.2 (M+H)+.
b) (+)-2,3,4,5-Tetrahydro-7-[[bis[(ben7imi-1~7nl-2-yl~methyl]amino]carbonyl]-3-oxo~-(2-phenethyl)- 1 H- l ,4-benzodiazepine-2-acetic 1.0 N LiOH (0.6 mL, 0.6 rnmol) was added dropwise at RT to a mixture of methyl (~)-2~3~4~5-tetrahydro-7-[[bis~(be~7~ 7nl-2-yl)methyl]amino]carbonyl]-3 oxo-4-(2-phenethyl)- 1 H- 1,4-benzodiazepine-2-acetate (0.13 g, 0.20 mmol) in THF
(5 mL) and H20 (5 rnL). After 18 h, the reaction mixture was concentrated to a small volume on the rotavap and cooled in an ice bath before neutralizing with 1.0 N
AcOH (0.6 mT.). The solution was Iyophilized to a crude product (0.092 g, 77~o) as .
W O 97/24119 PCTrUS96/20748 a white powder. ODS chromatography (step gradient, 5%-30%
CH3CN/H2O cont~inin?~ 0.1 % TFA) afforded the title compound as a white powder:
MS (ES) mle 628.2 (M+H)+.
Ex:~ml?le 34 Preparation of ethyl (~)-3-rrr2-(ben7imidazol-2-yl~ethyl~aminolsuccinoyllamino-4-pentynoate a) (+)-4-Ethynyl-2-azetidinone 4-Acetoxy-2-~7~ti(1inone (9.0 g, 69.7 mmol) was added slowly to a solution of ethynylm~gne~iumchloride (31 mL of a 0.5 M THF solution, 0.55 mmol) at 0~C.
After 1.5 h, 1.0 N HCl (100 mL) was added, and the mixture was taken up in EtOAc(300 mL) and washed sequentially with 1.0 N HCl (100 mL), saturated NaHCO3 (100 mL), and brine (100 mT.). After drying (MgSO4) and concentration, the titlecompound (4.57 g, 69%) was obtained as a light brownish solid: MS (ES) mJe 96.0 (M+H)+.
b) Ethyl (+)-3-amino-4-pentynoate A mixture of (_)-4-ethynyl-2-:~7~ tirlinnne (1.3 g, 13.68 mmol), EtOH (54 ml), and concentrated HCl (6 mL) was heated to reflux for 18 h. The reaction wascooled to RT before adjusting the pH to 8.0 using saturated NaHCO3. The reactionwas extracted with EtOAc (3 x 70 mL), and the combined EtOAc layers were washed with brine (50 ml). Drying (MgSO4) and concentration gave the title compound (1.06 g, 55%) as a brownish liquid: MS (ES) m/e 141.9 (M+H)+.
c) Methyl-[[2-(benzimid~ol-2-yl)ethyl]amino]succinate 3-Carbomethoxypropionylchloride(0.6 g, 4.0 mmol) was added at 0~C to a stirred solution of 2-aminoethylben7imi~ 701e ~ eet~te (1.13 g, 4.0 mmol) and diisol~lu~ylethylamine (2.59 g, 20 mmol) in dry CHzClz (45 rnL). After stirring for 1.5 h at RT, the reaction mixture was diluted with CH2Cl2 (50 mr ) and washed sequentially with H2O (30 mL), 5% NaHCO3 (30 ml), and brine (30 mL). Drying (MgSO4), concentration, and silica gel chromatography (8% MeOH/CH2C12) gave the title compound (0.2 g, 18%) as a yellow solid: MS (E~) m/e 276.4 (M+H)+.
. d) [[2-(Ben7imi~1~7-)1-2-yl)ethyl]amino]succinic acid W O 97/24119 PCT~US96/20748 A mixture of methyl-[[2-(ben7imi~ 7Ol-2-yl)ethyl]annino]succinate(0.2 g, 0.73 m~nol), 1.0 N NaOH (1.82 ml, 1.82 mmol) and MeOH (10 rnL) was stirred at RT for 24 h, then was concentrated to dryness. H2O
(5 mL) was added, the solution was neutralized with 1.0 N HCl (1.82 mL), and theresnlt;ng solution was lyophilized to give the crude title compound (0.23 g) as an off-white powder: MS (ES) m/e 261.9 (M+H)+.
e) Ethyl (_)-3-[~[2-(ben7.imi-1~7~ 2-yl)ethyl]amino]succinoyl]amino-4-pentynoateDiisopropylethylamine (0.32~mL, 1.83 mrnol) was added to a stirred solution of ethyl (+)-3-amino-4-pentynoate (0.12 g, 0.88 mmol), [[2-(be~ 7Ql-2-yl)ethyl]arnino]succinic acid (0.19 g, 0.73 mmol), HOBt H20 ~0.15 g, 1.1 mmol), and EDC (0.21 g, 1.1 mmol) in anhydrous CH3CN (15 ml) and DMF (3 mL) at RT.
After 23 h, the reaction mixture was concentrated, diluted with CH2Cl2 (100 mL),and washed sequentially with 5% NaHCO3 (2 x 25 mL) and brine (25 mL). Drying (MgSO4), concentration, and silica gel chromatography (7% MeOH/CH2Cl2) gave the title compound (0.07 g, 25%) as a white solid: MS (ES) mle 385.4 (M+H)+.
F.~mRle 35 P~ ~dlion of (_1-3-rrr2-(Ben7imitl~7ol-2-yl)ethyllaminolsuccinoyllamino-4 pentynoic acid a) (+)-3-r[r2-(Ben7imifi~7- 1-2-yl)ethyl]amino]succinoyl]amino-4-pentynoic acid 1.0 N LiOH (0.78 mL, 0.78 rnmol) was added dropwise at RT to a rnixture of ethyl (+)-3-[r[2-(bçn7imi~ 7ol-2-yl)ethyl]amino]succinoyl]amino-4-pentynoate (0.12 g, 0.31 mmol) in THF (5 mL), H2O (5 ml) and CH3CN (1 mL). After 3 h, the reaction mixture was concentrated to a small volume on the rotavap and cooled in an ice bath before neutralizing with 1.0 N AcOH (0.78 mL). The solution was lyophili7~l1 to a crude product (0.167 g) as a white powder. ODS chromatography (10% CH3CN/H2O cont~lining 0.1% TFA) afforded the title compound as a white powder: MS (ES) m/e 357.1 (M+H)+.
~;xample 36 Prep:lration of (+)-3-rrr4-(4-Azal~el.7i.. id~zol-2-yl)butanoyll~lycyllaminol-4-pentynoic acid (SB-237554) W O 97/24119 PCTrUS96/20748 a) Methyl (4-azabenzimidazol-2-yl)butyrate Triethylamine (319 mL, 22.9 mmol) was added to a mixture of 2,3-diaminopyridine (2.5 g, 22.9 mmol) and methyl 4-(chloroformyl)butyrate (3.77g, 22.9 mmol) in dry THF (50 mL) at 0~C. After stirring for 16 h at RT, the reaction was concentrated to dryness under vacuum. The residue was dissolved in glacial AcOH (25 mL) and was heated at 110~C. After 93 h, the reaction was allowed to - cool to RT and was conct;ntldL~;d under vacuum. The dark brown residue was diluted with H2O (40 mL) and CH2Cl2 (40 mL), and the mixture was neutralized to pH 7 using 5 N NaOH. The layers were separated and the aqueous layer was furtherextracted with CH2C12 (2x100 mL). The combined organic layers were washed sequentially with 5% NaHCO3 (2 x 30 mL)and brine (30 mL). Drying (MgSO4), concentration, and silica gel chromatography (7% MeOH/CH2Cl2) gave the title compound (0.47, 9%): MS (ES) m/e 220.0 (M+H)+.
~5 b) (4-Azabçn7imi(1zl7Ql-2-yl)butyric acid A lllii'~Ul'~ of methyl (4-:~7zlhen7imidazol-2-yl)butyrate (0.47 g, 2.13 mmol), 1.0 N NaOH (6 mL, 6.0 mmol) and MeOH (10 mL) was stirred at RT for 5.5 h, then was concentrated to dryness. The residue was diluted with H2O (2 mL) and neutralized with 1.0 N HCl (0.73 mL). The resulting solid was collected and air dried to give the title compound (0.32 g, 73%) as a yellow powder: MS (ES) m/e 206.0 (M+H)+.
c) Ethyl (_3-3-[[(N-tert-butoxycarbonyl)glycyl]amino]-4-pentanoate Diisopropylethylamine (0.92 mL, 5.32 mmol) was added to a stirred solution of ethyl I_)-3-amino-4-pentynoate (0.3 g, 2.13 mmol), Boc-Gly (0.56 g, 3.19 mmol), HOBt H2O (0.43 g, 3.19 mmol), and EDC (0.61 g, 3.19 mmol) in anhydrous CH3CN (15 mL) at RT. After 34 h, the reaction ll~Ult; was concentrated, diluted with CH2Cl2 (70 mL), and washed sequentially with 5% NaHCO3 (2x15 mL) and brine (15 mL). Drying (MgSO4), concentration, and silica gel chromatography (1:1EtOAc/Hexane) gave the title compound (0.5 g, 79%) as a colorless oil: MS (ES) m/e 299.2 (M+H)+.
d) Ethyl (:!~)-3-~(glycyl)amino}-4-pPnt~no~tl~ trifluoroacetate A solution of TFA (5 mL) and CH2Cl2 (15 mL) at RT was added all at once to ethyl (_)-3-~[(N-tert-butoxycarbonyl)glycyl]amino]-4-pe~t~no~te (0.5 g, 1.68 mmol). After 30 min, the solution was concentrated on the rotavap, and the residue W O 97/24119 PCTrUS96/20748 was reconcentrated from toluene (to remove residual TFA) to afford the title compound (0.55g, 106%) as a light yellow syrup: MS (ES) m/e 199.2 (M+H)+.
e) Ethyl (+)-3-~[[4-(4-azaben7imi~ 2-yl)butanoyl]glycyl]amino}-4-pentynoate Diisopropylethylamine (0.94 mL, 5.43 mmol) was added to a stirred solution of ethyl (+)-3-[(glycyl)amino]-4-pçnt~n-)~tlq trifluoro~l~et~te (0.55 g, 1.76 mmol), (4-azabenzimidazol-2-yl)butyric acid (0.32 g, 1.55 mmol), HOBt ~ H2O (0.31 g, 2.33 mmol), and EDC (0.45 g, 2.33 mmol) in anhydrous CH3CN (15 mL) at RT. After 64 h, the reaction mixture was concentrated, diluted with CH2Cl2 (100 mL), and washed sequentially with 5% NaHCO3 (2 x 25 mL) and brine (25 rnL). Drying (MgSO4), concentration, and silica gel chromatography (7% MeOH/CH2CI2) gave the title compound (0.11 g, 18%) as a white solid: MS (ES) m/e 386.4 (M+H)+.
f) (_)-3-[~[4-(4-Azaben7imi~1~7ol-2-yl)butanoyl]glycyl]amino]-4-pentynoic acid 1.0 N LiOH (0.71 mL, 0.71 mmol) was added dropwise at RT to a mixture of ethyl (+)-3-[[[4-(4-~abt n7imi~ 7ol-2-yl)butanoyl]glycyl~amino]-4-pentynoate (0.11 g, 0.285 mmol) in THF (5 mL), H2O (5 mL) and CH3CN (1 mL). After 2 h, the reaction mixture was concentrated to a small volume on the rotavap and cooled in an ice bath before neutralizing with 1.0 N AcOH (0.70 mL). The solution was lyophilized to a crude product (0.1 g, 100%) as a white powder. ODS
chromatography (5% CH3CN/H20 cont~ining 0.1% TFA) afforded the title compound as a white powder MS (ES) m/e 358.4 (M+H)+.
Fxz~m~?le 37 Preparation of (S)-2.3.4.5-tetrahydro-7-~rr(4-aza-$-methylben7irr i~ ol-2-yl)methyllmethylaminolcarbonyll-3-oxo- lH- 1.4-benzodiazepine-2-acetic acid a) Dimethyl D-malate-o-trifluorom~th~npslllfonate A solution of dimethyl-D-malate (12.96 g, 80 mmol) and pyridine (6.8 mT., 84 mmol) in CH2Cl~ dry (50 mL) was added dropwise under argon at 0~C to a solution of trifluo~ nesulfonic anhydride (14.2 rnL, 84 mmol) in dry CH2Cl2 (40 rnL) in a flame dried flask. The resulting yellowish-orange mixture was stirred at 0~C for 30 rnin, and then at RT for 4 hr. The reaction was qllen~h~l by adding H20 (50 rnL), and the layers were separated. The organic layer was washed sequentially with H2O (3 x) and brine. Drying (MgS04) and concentration gave thetitle compound (22.45 g, 95%) as an off white solid: MS(ES) m/e 295 (M + H)+.
CA 0224l633 l998-06-26 W O 97/24119 PCTrUS96/20748 b) Dimethyl-N-(2-cyanophenyl)-D-aspartate A solution of dimethyl D-malate-O-trifluo~ n--sulfonate (22.4 g, 76.2 mmol) in CHCl3 (40 mL) and hexanes (40 mL) was added at to a solution of 2-aminobenzonitrile (9.0 g, 76.2 mmol) and 2,6-di-tert-butylpyridine in CHCl3 (50 mT) and hexanes (50 mL) in a flame dried flask at 0~C under argon. The resulting- mixture was stirred at 0~C for 30 min, then at RT for 3 days. The solvent was removed in vacuo and the residue was taken into EtOAc and washed sequentially with 5% HCI (10 x) and brine. Drying (MgSO4), concentration and silica gel flashchromatography (12% EtOAc/hexanes) gave the title compound (12.3 g, 62%) as a clear oil: MS(ES) mte 263.3 (M + H)+.
c) Methyl (S)-2,3,4,5-tetrahydro-3-oxo- lH- 1 ,4-benzodiazepine-2-acetate A mixture of dimethyl-N-(2-cyanophenyl)-D-aspartate ~12 g, 45.7 mmol), Et3N (7.64 mL, 54.84 mmol) and Raney-Ni (46 g, wet, prewashed with CH30H) in CH30H (200 mL) was stirred at RT under H2 (balloon) for 2 days. The catalyst wasremoved by filtration and washed with CH30H (3 x). Concentration and silica gel flash chromatography (0-5% CH3OH/CH2Cl2) gave the title compound (7.93 g, 74 %) as a white solid: MS(ES) m/e 235.3 (M + H)+.
d) Methyl (S)-2,3,4,5-tetrahydro-7-bromo-3-oxo- 1 H- 1 ,4-benzodiazepine-2-acetate Tetrabutylammonium tribromide (5.16 g, 10.7 mmol) was added portionwise to a solution of methyl (S)-2,3,4,5-tetrahydro-3-oxo-lH-1,4-benzodiazepine-2-acetate (2.5 g, 10.7 mmol) in CHCl3 (50 mL), and the llliY~tUl'e was stirred at RT for 2 days. H2O (30 mL) was then added, and the organic layer was separated and washedsequentially with H20 and brine. Drying (MgSO~), concentration, and silica gel flash chromatography (0-5% CH3OH/CH2Cl2) gave the title compound (1.99 g, 60%)) as a white solid: MS(ES) m/e 313.0 (M + H)+.
e) Methyl (S)-2,3,4,5-tetrahydro-7-[t[(4-aza-5-methylbeu7i.l.i~l~7Ol-2-yl)methyl]methylamino]carbonyl]-3-oxo- 1 H- 1 ,4-benzodiazepine-2-acetate A ~ tule cont~inin~ methyl (S)-2,3,4,5-tetrahydro-7-bromo-3-oxo-lH-1,4-benzodiazepine-2-acetate (624 mg, 2 mmol), 2-(aminomethyl)4-a_a-5-methylben7imicl~7ole dihydrochloride (695 mg, 2.8 mmol), DIEA (1.8 mL, 10 mmol), and (Ph3P)2PdCl2 (126 mg, 0.18 mrnol) in NMP (22 mL) was heated to 110~C under a CO balloon for 48 hr. The solvent was removed on the rotavap (highvacuum) and the residue was puri~led by silica gel flash chromatography (0.5 - 5%
W O 97/24119 PCTrUS96120748 CH3OEI/CH2Cl2) to give the title compound (170 mg, 19.5%) as a pale yellow solid: MS (ES) m/e 437.5 (M + H)'.
f~ (S)-2,3,4,5-Tetrahydro-7-[~[(4-aza-5-methylben7imitlz-~ol-2-5 yl)methyl]methylamino]carbonyl]-3-oxo- 1 H- 1,4-benzodiazepine-2-acetic acid 1.0 M LiOH (0.6 mL, 0.6 mmol) was added dropwise to a solution of methyl (S)-2,3,4,5-tetrahydro-7-[[[(4-aza-5-methylbçn~.imi~1~7.ol-2- -yl)methyl}methylamino]carbonyl]-3-oxo-lH- 1,4-benzodiazepine-2-acetate (170 mg, 0.39 rnmol) in CH30H (5 mL) and THF (5 mL)at RT. The resulting rnixture was 10 stirred for 20 hr and then was concenlldted. The residue was dissolved in H20, acidified with 30% TFA, and purified by ODS chromatography (5% CH3CN/H20 contz-ining 0.1% TFA). Concentration and lyophilization gave the title compound as an off white powder: [a],,25 -74.5o (c = 1, CH30H); MS (ES) m/e 423.2 (M + H)~.
Anal. Calcd for C2,H22N604 ~ 2 TFA ~ 1.75 H20: C, 44.03; H, 4.06; N, 12.32. Found:
C, 44.33; H, 4.04; N, 12.28.
Fx~nlple 38 Plep~dlion of (S)-2.3~4.5-tetrahydro-7-rrr(ben7imitl~zo1-2-yl)metbyl~minolcarbonyll-3-oxo- 1 H- 1 ~4-benzodiazepine-2-acetic acid a) Methyl (S)-2,3,4,5-tetrahydro-7-iodo-3-oxo- l H- 1,4-benzodiazepine-2-acetatePyridine-ICI complex: Iodine monochloride (100 mL, 1 M solution in CH2Cl2) was added slowly to a solution of pyridine (8.5 mL, 105 mmol) in dry CH2Cl2 (20 mL) at 5~C under argon so that the temperature was m~int~in~d between10 - 15~C. The mixture was stirred at 5 - 10~C for 20 min, then hexanes (50 mL) was added, and the rnixture was stirred in the cold bath for another 30 min. Thesolid was collected by suction filtration, washed sequentially with hexanes and petroleum ether, and dried to afford the reagent (22.5 g) as a yellow, crystalline solid.
Pyridine-ICl complex (1.27 g, 5.28 mmol) was added portionwise to a solution of methyl (S)-2,3,4,5-tetrahydro-3-oxo-lH-1.4-benzodiazepine-2-acetate (1.18 g, 4.8 mmol) in CH2Cl2 (20 rnL) and CH30H ~20 mT). The resulting mixture was stirred at RT for 40 rnin, then l M NaHSO3 (20 mL) was added. The solid was collected by suction filtration and washed with Et2O. Drying yielded the title compound (1.72 g, qu~ntit~tive) as an off-white solid: MS (ES) m/e 361.2 (M + H)~.
b) Methyl (S)-2,3,4,5-tetrahydro-7-r[[(ben7imi~ 7OI-2-yl)methyl]amino]carbonyl]-3-oxo- l H- 1,4-benzo~ 7~pine-2-acetate A mixture of methyl (S)-2,3,4,5-tetrahydro-7-iodo-3-oxo-lH-1,4-benzodia_epine-2-acetate (1.08 g, 3 mmol), 2-aminomethylben7imi~1~7Ole S dihydrochloride hydrate (924 mg, 4.2 mmol), DIEA (2.6 mL, 15 mmol), and (Ph3P)2PdCl2 (211 mg, 0.3 mmol) in NMP (30 mL) was heated to 110~C under a CO
- balloon for 3 hr. The solvent was removed on the rotavap (high vacuum) and the residue was purified by silica gel flash chromatography (0-7% CH30H/CH2Cl2) to afford the title compound (530 mg, ~44%) as an off white solid: MS(ES) m/e 408.1 (M+H)t.
c) (S)-2,3,4,5-Tetrahydro-7-{[[(ben7imi~ 7ol-2-yl)methyl]amino]carbonyl]-3-oxo-1 H- 1,4-benzodiazepine-2-acetic acid Following the procedure of Example 37(f), except substituting methyl (S)-2,3,4,5-tetrahydro-7-[[[(ben7imi(1z~7Ol-2-yl)methyl]amino]carbonyl]-3-oxo-lH-1,4-benzodiazepine-2-acetate for methyl (S)-2,3,4,5-tetrahydro-7-[[[(4-aza-5-methylbenzimidazol-2-yl)methyl]methylamino3carbonyl]-3-oxo- lH- 1,4-benzodiazepine-2-acetate, the title compound (66%) was ~ al~,d as a white powder: [a]n25 -145.3~ (c = 1, CH30H); MS (ES) m/e 394.2 (M + H)t. Anal. Calcd for C2nH,gN5O4 - 2 TFA 0.125 H2O: C, 46.22; H, 3.43; N, 11.23. Found: C, 46.13;
H, 3.78; N, 11.49.
E~am~l?le 39 ~lGpaldlion of (+)-N-r2-(aminomethyl~-4-rrr~4-~7~-S-methylbe~7,imi-1~7ol-2-yl)methyl~methylaminolcarbonyllphenyl~aspartic acid a) Methyl (+)-2,3,4,5-tetrahydro-7-[[[(4-a_a-5-methylben7imi~1~7ol-2-yl)methyl]methylamino]carbonyl3 -3-oxo- 1 H- 1,4-benzodia_epine-2-acetate EDC (130 mg, 0.75 mmol) was added to a solution of methyl (+)-2,3,4,5-tetrahydro-7-carboxy-3-oxo-lH-1,4-benzodiazepine-2-acetate (190 mg, 0.68 mmol), 2-(aminomethyl)-4-aza-S-methylben7imi~l~7ole dihydrochloride (169 mg, 0.68 mmol), HOBt H2O (101 mg, 0. 75 mmol), and DIEA (0.39 mL. 2.24 mmol) in anhydrous DMF at RT. After 20 hr, the reaction was concentrated on the rotavap (high vacuum), and the residue was cl~ atographed on silica gel (1 - 6.5%
CH3OH/CH2Cl2) to afford the title compound (260 mg, 88%) as a white solid: MS
(ES) m/e 437.5 (M ~ H)'.
b) (_)-N-[2-(Aminomethyl)-4-[[[(4-aza-5-methylben7imitl~7O1-2-yl)methyl]methylamino}carbonyl]phenyl]aspartic acid Following the procedure of Example 37(f), except substitl-ting methyl (+)-2,3,4,5-tetrahydro-7-[[[(4-~a-5-methylben7imi~1~7O1-2-yl)methyl]methylamino]carbonyl]-3-oxo-lH-1,4-ben7O~ 7~pine-2-acetate for methyl (S)-2,3,4,5-tetrahydro-7-[[[(4-aza-5-methylben7imi~1~701-2-yl)methyl]methylamino]carbonyl]-3-oxo- 1 H- l ,4-bçn70~ 7epine-2-acetate, the title compound (66%) was prepared as a white powder: MS (ES) m/e 441.2 (M + H)+.
Anal. Calcd for C2,H2"N6O5 - 2 TFA 2.25 H2O: C, 42.08; H, 4.38; N, 11.78. Found:C, 42.01; H, 4.18; N, 11.55.
Fx~rru?le 40 Plepaldlion of (S~-2~3.4.5-tetrahydro-7-lrr(4-aza-5-methylbell7i.. i~zl7ol-2-yl)mt~thyll~minolcarbQ~.~yll-3-oxo-lH-1~4-benzodiazepine-2-acetic acid a) Methyl (S)-2,3,4,5-tetrahydro-7-[[[(4-aza-5-methylberl 7i mi~l ,.7 ol-2-yl)methyl]amino]carbonyl]-3-oxo- I H- 1,4-benzodiazepine-2-acetate Following the procedure of Example 38(b), except ~7u1)~7LiLu~il.g 2-(aminomethyl)-4-aza-5-methylbell,i..,i-lz~7Ole dihydrochl(-ride for 2-aminomethylbe~7imicl~7ole dihydrochloride hydrate, the title compound (63%) was prepared as an amber solid: MS ~ES) m/e 423 (M + H)'.
b) (S)-2,3,4,5-Tetrahydro-7-[[[(4-aza-5-methylben7imicl~7O1-2-yl)methyl]amino]carbonyl~-3-oxo- 1 H- 1,4-benzodiazepine-2-acetic acid Following the procedure of Example 37(f), except substituting methyl (S)-2,3,4,5-tetrahydro-7-[[[(4-aza-5-methylb~n7imi~ 7ol-2-yl)methyl]amino]carbonyl]-3-oxo- lH- 1,4-benzodiazepine-2-acetate for methyl (S)-2,3,4,5-tetrahydro-7-t[[(4-aza-5-methylbe~-7i.. i-l~7~-1-2-yl)methyl]methylamino]~ul,o.,yl]-3-oxo-lH-l,~benzodiazepine-2-acetate, the title colll~oulld (50%) was prepared as a white powder: MS (~S) m/e 409.2 (M + H)~. Anal. Calcd for C2"H2"N6O4 ~ 1.75 TFA -H~O: C, 45.09; H, 3.82; N,3 .45. Found: C, 45.18; H, 4.10; N, 13.58.
E~mple 41 W O 97/24119 PCTrUS96/20748 Preparation of (S)-2.3.4.5-Tetrahydro-7-rrr(bPnzimitl:~7ol-2-yl)methyllaminolcarbonyll-3-oxo-4-r2-(pyrid-3-yl)ethyll-lH- 1.4-ben7-1diazepine-2-acetate 5 a) tert-Butyl 4-fluoro-3-[[2-(pyrid-3-yl)ethyl~amino]benzoate A mixture of tert-butyl 4-fluoro-3-methylbenzoate (3.83 g, 18.22 mmole), - NBS (3.57 g, 20.24 mmole), benzoyl peroxide (0.22 g, 0.91 mmole), and CC14 (90 mL) was heated at reilux. After 16 hr, the reaction was cooled in ice/H20 and filtered, and the filtrate was concentrated. The residue was passed through a short 10 pad of silica gel (20 % EtOAc/hexane) to remove baseline m~t~.ri~ls~ and the filtrate was concentrated. The residue was dissolved in THF ~90 mI,), and 3-(2-aminoethyl)pyridine (6.97 g, 57 mmol).) was added rapidly. The addition a~eal ,dto be mildly endothermic. The reaction was stirred overnight then was concentrated.
The residue was diluted with Et2O (100 mL) and washed sequentially with 1.0 N
15 NaOH (30 mL), H2O (30 mT.), and brine (30 mL). Drying (MgSO4), concentration,and silica gel chromatography (10 % MeOH/CH2Cl2) gave the title compound (2.58 g, 59 %) as a yellow oil: MS (ES) m/e 331 (M + H)+.
b) tert-Butyl (S)-4-fluoro-3-[2-aza-4-(benzyloxycarbonyl)amino-3,6-dioxo-6-20 methoxy-2-[2-(pyrid-3-yl)ethyl]hexyl]benzoate DCC (1.86 g, 9 mmol)was added to a solution of tert-butyl 4-fluoro-3-[[2-(pyrid-3-yl)ethyl]amino]benzoate (2.7 g,.8.18 mmol), N-Cbz-L-aspartic acid ~-methyl ester (J. Am. Chem. Soc. l9S7, 79, 5697; 2.53 g, 9 mmol.), and HOBt H2O
(1.2 g, 9 mmol) in anhydrous DMF (10 mL) at RT. After 24 hr, the mixture was 25 diluted with Et20 (25 rnL) and filtered. The filtrate was concentrated to dryness, and the residue was diluted with Et2O (50 rnL) and washed with H2O (2 x 10 mL) and brine (10 mL). Drying (MgSO4), concentration, and silica gel chromatography (CH2Cl2) gave the title compound (2.4 g, 49%)as a colorless oil: MS (ES) m/e 594(M +H)+.
c) tert-Butyl (S)-4-fluoro-3-[4-amino-2-aza-3,6-dioxo-6-methoxy-2-[2-(pyrid-3-yl)ethyl]hexyl]benzoate A mixture of tert-butyl (S)-4-fluoro-3-[2-aza-4-(benzyloxycarbonyl)arnino-3,6-dioxo-6-methoxy-2-[2-(pyrid-3-yl)ethyl]hexyl]benzoate (2.4 g, 4 mmol.), 10 %35 Pd/C (184 mg, 0.17 mmole), and MeOH (17 mL) was shaken at RT under H2 (50 psi). After 1.5 hr, the reaction was filtered through celite(~3) and concentrated. Silica W O 97/24119 PCT~US96/20748 gel chromatography (10 % MeOH in 1:1 EtOAc/CHC13) gave the title compound (1.1 g, 59 %) as a colorless oil: MS (ES) 460 (M + H)+.
d) Methyl (S)-2,3,4,5-tetrahydro-7-(tert-butoxycarbonyl)-4-[2-(pyrid-3-yl)ethyl]-3-5 oxo- lH- 1,4-benzodiazepine-2-acetate A solution of tert-butyl (S)-4-fluoro-3-t4-amino-2-aza-3,6-dioxo-6-methoxy-2-[2-(pyrid-3-yl)ethyl]hexyl]ben70~te (0.64 g 1.39 mrnol) in anhydrous DMSO (5.7mL) was heated under argon in an oil bath set at 120-125~C. After 17.5 hr, the reaction was cooled in ice/H20 and~diluted with H20 (12 mL). The mixture was 10 extracted with EtOAc (3 x 20 mL), and the combined EtOAc layers were washed with H2O (10 mL) and brine (10 mL). Drying (MgSO4), concentration, and silica gel chromatography (9: 1 CH2Cl2/MeOH) gave the title compound (0.15 g, 33%) as anearly colorless solid: MS (ES) m/e 440 (M +H)+.
e) Methyl (S)-2,3,4,5-tetrahydro-7-carboxy-4-[2-(pyrid-3-yl)ethyl]-3-oxo-lH-1,4-benzodiazepine-2-acetate 4 M HCI/dioxane (0.5 mL) was added to a solution of methyl (~)-2,3,4,5-tetrahydro-7-(tert-butoxycarbonyl)-4- [2-(pyrid-3-yl)ethyl]-3 -oxo- 1 H- 1,4- -benzodiazepine-2-acetate(0.18 g, 4.1 mrnol) in anhydrous CH2Cl2 (5 mL) and the reaction was stirred at RT overnight. Concentration in vacuo followed by reconcentration from toluene (3 x 10 mL) gave the title compound (0.12 g, 65%):
MS (ES) m/e 384 (M + H)+.
f) Methyl (S)-2,3,4,5-tetrahydro-7-[t[(ber 7imitl~7ol-2-yl)methyl]amino]carbonyl]-3-oxo-4-[2-(pyrid-3-yl)ethyl]- 1 H- 1,4-benzodiazepine-2-acetate A mixture of methyl (S)-2,3,4,5-tetrahydro-7-carboxy-4-[2-(pyrid-3-yl)ethyl]-3-oxo-lH-1,4-benzo~ 7epine-2-acetate (0.12 g, 0.26 mmol) and thionyl chloride (15 mL) was refluxed for 1 h. The re~slllting orange solution was concentrated to dryness to leave a yellow-orange foam. This was dissolved in CH2C12 (10 mL) and added dropwise to a solution cont~inin~ 2-aminomethylbe..~ilni~l~701e dihydrochloride hydrate (0.058 g, 0.26 mmol), pyridine (0.72 g, 9.1 mmol), and triethylamine (0.55 g, 5.46 mmol) in CH2C12 (15 mL) at 0~C under argon. The reaction mixture was then stirred in RT under argon. After 25.5 h, CH2C12 (200 mL) and 5% NaHCO3 (50 rnL) were added to the reaction 35 mixture to give a light yellow precipitate which was filtered and air-dried to give the title compound (0.030g.,~% yield): MS (ES) m/e 513 (M+H)+.
W O 97/24119 PCTrUS96/20748 g) (s)-2~3~4~5-Tekahydro-7-~[[(ben~ 7ol-2-yl)methyl]amino]carbonyl] -3-oxo-4-[2-(pyrid-3-yl)ethyl]- 1 H- 1,4-benzodiazepine-2-acetate 1.0 N LiOH (0.57 mL, 0.57 mmol) was added dropwise at RT to a mixture of S methyl (S)-2,3,4,5-tetrahydro-7-[[[(ben7imi(1~7~-1-2-yl)methyl3amino]carbonyl]-3-oxo-4-[2-(pyrid-3-yl)ethyl]-lH-1,4-benzodia_epine-2-acetate (0.030 g, 0.059 mmol) - in THF (4 mL) and H2O (5 mL). The resulting light brownish-yellow solution was stirred for 21.5 h, then was concenkated on the rotavap. The resulting residue was lyophilized to give the crude product as a yellowish powder. ~ liv~ HPLC
(PRP-l~ column, step gradient, 10-20% CH3CN/H2O contzlinin~ 0.1% TFA) afforded the title compound (0.OlOg 34% yield): MS (ES) m/e 499(M+H)+. Anal.
Calcd .for C27H26N6O4 - 3 C2HF3O2 ~ 3 HCI - 3 H2O: C, 37.41; H, 3.41; N,7.52.
Found: C, 37.6; H, 3.52; N, 7.52.
Example 42 Preparation of Ethyl (S)-2.3.4.5-tetrahydro-7-rrr~ben7imid~7ol-2-yl)methyl~methylaminolcarbonyll-4-methyl-3-oxo- 1 H- 1.4-benzodiazepine-2-acetate a) Ethyl (s)-2~3~4~s-tekahydro-7-~[[(be~ 7~l-2-yl)methyl]methylamino]carbonyl]-4-methyl-3-oxo- lH- 1,4-benzodia_epine-2-acetateHCl gas was bubbled into EtOH (200 mL) for 10 min, then (S)-2,3,4,5-tetrahydro-7-[[[(ben7imi~ 7ol-2-yl)methyl]methylamino]carbonyl]~-methyl-3-oxo-lH-1,4-benzodia7epine-2-acetic acid (2.00 g, 4.5 mmol) was added. The reaction was stirred at RT for 24 hr, then was concentrated to dryness on the rotavap. The residue was reconcenkated from toluene (2 x) to remove residual EtOH, then was chromatographed on silica gel (gradient: 7% MeOH/CH2Cl2 (1 L) then 10%
MeOH/CH2Cl2). The r~sllltin~ residue was dissolved in EtOH, and Et2O was added to precipitate a solid. This was collected and washed with Et2O to afford the title compound as a white solid: MS (ES) m/e 450.2 (M + H)+. Anal. Calcd for C24H27NsO4 ~ 1.5 H2O: C, 60.49; H, 6.35; N, 14.70. Found: C, 60.41; H, 6.27; N, 14.38.
Fx~mple 43 Preparation of 4-~rr3-(lH-ben7imi~1~7ol-2-yl)propyllaminolcarbonyllpiperidine-l-acetic acid W O 97/24119 PCTrUS96/20748 a~ 2-[3-~N-tert-Butoxycarbonyl)aminopropyl3benzirnidazole A solution of isobutylchloroformate ( 10.2 rnL, 79 mmol) in THF (25 rnL) was added to a solution of 4-(tert-butoxycarbonyl)arninobutyric acid (Organic S ~;ynthesis 1984, 63, 160; 13.5 g, 0.066 mole) and triethylamine (11 mL, 80 mmol) in THF ( 50 mL) at 0~C under argon. After 0.5 h, a solution of 1,2-phenylen~ mine (7 g, 64.8 rnmole) in THF (50 rnL) was added dropwise to the res-ltting white suspension. The reaction was stirred for 18 h, then was ffltered, and the filtrate was concentrated to give a serni-solid. This was dissolved in AcOH (100 rnL), and the solution was heated at 70~C for 18 h. The reaction mixture was concentrated, andthe residue was reconcentrated several times from toluene. Silica gel flash chromatography gave the title compound (6.0 g, 33%): MS (ES) m/e 276 [M+H]t.
b) 2-(3-Aminopropyl)bc~.~i...itl~7ole dihydrochloride A solution of 2-[3-(N-tert-butoxycarbonyl)aminopropyl3ben7imirls-7.nle (1.2 g, 4.3 mmol) and 4 M HCl in dioxane (20 mL) in CH2C12 (25 ml) was stirred at RT
for 18 h. The resulting white suspension was filtered to give the title compound(1.07 g, 97%).
20 c) Ethyl ~[[[3-(lH-ben,.iid~7.ol-2-yl)propyl]arnino]carbonyl]piperidine-l-acetate A ~ lule of ethyl 4-carb-~y~ 3elidine-1-acetate hydrochloride (Yellin's SB
223913 CIP)(0.76 g, 3 mmol) in thionyl chloride (10 mL) was heated to reflux for15 min, then was concentrated to dryness. After evaporation several times from toluene, the residue was dissolved with 2-(3-aminopropyl)ben7imi~1~7Ole dihydrochloride (0.77 g, 3 mmol) and DIEA (3 mL) in DMF (25 mL). After 18 h, the reaction mixture was partitioned between EtOAc (50 mL) and 5% NaHCO3 (100 mL), and extracted with EtOAc (2 x 50 mL). The combined organic extracts were washed seq~lenti:~ily with H20 and saturated NaCl solution, then were dried overMgSO4. Concentration and silica gel flash chromatography (6% MeOH/CH2C12) gave the title compound (40 mg, 3%).
b) 4-[[~3-(Rçn7.imida_ol-2-yl)propyl]amino~carbonyl]piperidine-1-acetic acid lN NaOH solution (0.4 mL, 0.4 mmol) was added to a stirred solution of ethyl 4-[[3-~lH-ben7imi~l~7ol-2-yl)propylamino]carbonyl]piperidine-l-acetate (40mg, 0.1 mmol) in MeOH (10 mL)at RT. After 18 h, the mixture was neutralized with AcOH (1 mL) and concentrated to remove the MeOH. The aqueous solution was loaded onto an XAD-2 column, and eluted with H20 (500 mL), then with 20%
W O 97/24119 PCTrUS96/2074P
CH3CNt H20. The fractions con,~zlining the product were pooled and lyophilized to give the title compound (9 mg, 25%). MS (ES~ m/e 345.2 [M+H]+.
Anal. Calcd for C,8H24N4O3 0.75 H2O: C, 60.40; H, 7.18; N, 15.65. Found: C, 60.48; H, 7.16; N, 15.40.
Lxample 44 Plc~aldliQn of 4-rrr3-(ben7imi~l~7f 1-2-yl)propyllaminolcarbonyllphenylacetic acid a) Ethyl 4-[[[3-~ben7imiclz~ 1-2-yl)propyl]amino]carbonyl]-l-phenylacetate A mixture of ethyl 2-(4-carboxyphenyl)acetate (Yellin's SB 223913 CIP)(0.5 g, 2.4 mmol), 2-(3-aminopropyl)ben7imi~7ole dihydrochloride (0.7 g, 2.8 mmol), HOBt ~ H20 ~0.36 g, 2.6 mmol), EDC (0.5 g, 2.6 mmol), and DIEA (1.5 mL, 8.8 mmol) in DMF (15 mL) was warmed briefly and stirred at RT for 18 h. The reactionmixture was partitioned between EtOAc ~50 mL) and 5% NaHCO3 (100 mL), and extracted with EtOAc (2 x 50 mL). The combined organic extracts were washed sequentially with H2O and saturated NaCI solution, then were dried over MgSO4.
The evaporated residual solid was triturated with Et2O to give the title compound (0.56 g, 66%): MS (ES) m/e 366.0[M+H~+. Anal. Calcd for C2lH23N3O3 0.25 H2O:
C, 68.18; H,6.40; N, 11.36. Found: C, 68.16; H,6.26; N,11.36.
b) 4-[[[3-(Ben7imi~1~7(-1-2-yl)propyl]amino]carbonyl]phenylacetic acid 1 N NaOH solution (6 rnL, 6 mmol) was added to a stirred solution of ethyl 4-[[[3-(lH-bPn7imicl~7n-2-~71)propyl]arnino]carbonyl]-l-phenylacetate (0.38 g, 1 25 mmol) in MeOH (15 mL)at RT. After 4 h, the mixture was neutralized with AcOH
(6 mL) and the resulting solid was filtered to give the title compound (77 mg, 22%):
MP 108-110~C; MS (ES) rn/e 345.2 [M+H]+. Anal. Calcd for Cl9HlgN3O3 0.6 H2O:
C, 65.54; H, 5.85; N, 12.07. Found: C, 65.63; H, 5.65; N, 11.95.
Fx~m~le 45 Preparation of (S)-2.3.4.5-tetrahydro-4-m~t~yl-3-oxo-7-rrr(S-trifluoromethylben~ 7ol-2-y~)methyl~methylaminolcarbonyll - l H- l .4-benzodiazepine-2-acetic acid a) 3,4-Diaminobenzotrifluoride W O 97~4119 PCT~US96/20748 4-Amino-3-nitroben~oLlinuoride (3.7070 g, 17.98 mmol) was dissolved in MeOH, and a catalytic amount of 10% Pd/C was added. The reaction was purged with H2, and stirred at RT under H2 (balloon). After 24 hr, the reaction was filtered through a bed of celite~, and the filtrate was evaporated under vacuum to yield the title compound (3.0878 g, 98%). The material was used without characlefizdLion.
b) 2-[N-(Benzyloxycarbonyl)-N-methyl]aminomethyl-5-trifluoromethylben7imi~ 1e Cbz-sarcosine (3.9950 g, 17.13 mmol) was dissolved in dry THF, and isobutylchloroformate (2.5 mL, 19.27 mmol) was added, followed by triethylamine (5.0 mL, 35.95 mmol). The mixed anhydride was allowed to form at RT for 30 min, then was added to a solution of 3,4-diaminobenzotrifluoride (3.0818 g, 7.53 mmol) in dry THF. After 20 hours at RT, the reaction was ~vapoldted under vacuum. The residue was partitioned between EtOAc and 1.0 N NaHCO3, and the layers were separated. The aqueous layer was extracted with EtOAc, and the combined organic layers were dried (MgSO4), filtered and e-v~oldled under vacuum. The residue wasreconcentrated from toluene, then was dissolved in glacial AcOH (125 mL). The solution was heated at 110~C for 24 hr, then the AcOH was evaporated under vacuum. The residue was reconcentrated from toluene, then was adsorbed onto silica gel and loaded onto a dry silica gel flash chromatography column. The column was eluted with 1:1 CHCl3/Et,0 to afford the title compound (2.9397 g, 47.2%): TLC R, (1:1 CH2Cl~/Et2O) 0.57; MS (ES) rn/e 364.2 (M+H)+; 'H NMR (250 MHz, CDC13) ~ 8.0 - 7.2 (m, 9H), 5.05 (s, 2H), 4.84 (s, 2H), 3.07 (s, 3H).
c) 2-(Methylaminomethyl)-5-trifluoromethylben7.imicl~701e 2-[N-(Benzyloxycarbonyl)-N-methyl]aminomethyl-S-tri~uoromethylben7imi(1~7ole (2.9397 g, 8.09 mmol) was dissolved in MeOH, and a catalytic amount of 10% Pd/C was added. The reaction was purged with H2, then was stirred at RT under H2. After 5 hr, the reaction ~ lure was filtered through a bed of celite(~), and the filtrate was evaporated under vacuum to leave a tan colored oil. Analysis by 400 MHz NMR showed the Cbz protecting group was still present, so the residue was resubmitted to the reaction conditions. After 18 hr, the catalyst was removed by filtration through a bed of celite(~) and the filtrate was ~;v~oldted under vacuum to yield the title compound (1.7809 g, 96%): lH NMR (250 MHz, CDC13) o 7.76 - 7.32 (m, 4H), 4.32 (s, 2H), 2.59 (s, 3H).
W O 97/24119 PCT~US96/20748 d) Methyl (S)-2,3,4,5-tetrahydro4-methyl-3-oxo-7-[[[(5-trifluoromethylben7im~ 7Ol-2-yl)methyl]methylamino]carbonyl]- lH- 1,4-benzodiazepine-2-acetate Methyl (S)-7-carboxy-4-methyl-3-oxo-2,3 ,4,5-tetrahydro- l~I- 1,4-S benzodiazepine-2-acetate (179.2 mg, 0.61 mmol) was weighed into a 100 mL round bottomed flask. CH3CN (10 mL) was added, followed by HOBt ~ H2O (97.9 mg, 0.72 mmol) and EDC (149.3 mg, 0.78 mmol). After all the solids had dissolved, a solution of 2-(methylarninomethyl)-5-trifluoromethylben7imi~1~701e (186.1 mg, 0.81 mmol) in CH3CN was added with diisopropylethylamine (0.25 rnL, 1.44 mmol).
After 24 hr at RT, the reaction was evaporated under vacuum, and the residue waschromatographed on silica gel (3% MeOH/CHCl3) to afford the title compound (308.1 mg, 100%): TLC Rf (5% MeOH/CHCl3) 0.21; lH NMR (250 MHz, CDC13) 7.83-7.16 (m, 7H), 5.37 (d, 1~), 5.05-4.70 (m, 3H), 2.96 (m, 3H), 3.72 (s, 3H), 3.16 (s, 2H), 2.11 (s, 3H); MS (ES) m/e 504.0 (M+H)~.
e) (s)-2~3~4~5-Tetrahydro-4-methyl-3-oxo-7-[[[(s-trifluoromethylben~ 7Q1-2 yl)methyl]methylamino]carbonyl] -1 H- 1 ,4-benzodiazepine-2-acetic acid Methyl (S)-2,3,4,5-tetrahydro-4-methyl-3-oxo-7-[[[(5-trifluoromethylb~n7imi~1~7ol-2-yl)methyl]methylamino~carbonyl]-lH-1,4-benzodiazepine-2-acetate (308.1 mg, 0.61 mmol) was dissolved in MeOH (S mL).
H20 (5 mL) was added, followed by 1.0 N NaOH (2.0 mL, 2.0 mmol). After 24 hr at RT, the reaction was neutralized with 1.0 N HCl (2.0 mL). The milky white mixture was stirred at RT for 15 min, then was diluted with H2O, and the precipitate was collecte~l on a sintered glass funnel. The white powder was dried in a vacuum de~ c~tQr overnight to yield the title compound (268.0 mg, 90%): M~ (ES) m/e 490.2 (M+H)'. Anal. Calcd for C23H~N5O4F3 2.25 H2O ~ 0.25 HCl: C, 51.24; H, 5.00; N, 12.99. Found: C, 51.44; H, 4.96; N, 12.45.
F.x:~ml?le 46 Ple~ alio l of (S)-2.3.4.5-tetrahydro-7-rrr(4.7-dimethoxyben7imicl~7.ol-2-yl)methyllmethylamino~carbonyll-4-methyl-3-oxo- lH- 1 .4-benzodiazepine-2-aceticacid ~' a) 2-[N-(Benzyloxycarbonyl)-N-methyl]~n~innmPthyl-4,7--1im~thoxybenzimidazole ~ Following the procedure of Example 45(b), except sul)~lilulillg 1,2-~ mino-3,6-dimethoxybenzene for the 3,4-diaminobenzotrifluoride, the title compound was 167 W O 97~4119 PCT~US96~0748 prepared: MS (ES) m/e 356.2 (M+H)'; lH NMR (250 MHz, CDC13) o 7.34 (s, SH), 6.54 (d, 2H), 5.18 (s, 2H), 4.65 (s, 2H), 3.95 (s, 3H), 3.86 (s, 3H), 3.03 (s,3H).
S b) 4,7-Dimethoxy-2-(methylaminomethyl)ben7imi(1~7ole 2-[N-(Benzyloxycarbonyl)-N-methyl]aminomethyl-4,7-dimethoxybenzimidazole (186.5 mg, 0.53 mmol) was dissolved in MeOH, and a catalytic amount of 10% Pd/C was added. The reaction was purged with Hz, then was stirred at RT under H~ (balloon). ~After 20 hr, the reaction was filtered through celite(~, and the filtrate was eva~?oraLed under vacuum to yield the title compound (96.9 mg, 83%): 'H NMR (250 MHz, CDC13) ~ 6.52 ~s, 2H), 3.94-3.86 (m, 6H), 2.36 (s, 3H).
c) Methyl (S)-2,3,4,5-tetrahydro-7-[[[(4,7-~lim~thQxyben7imifl~7ol-2-yl)methyl]methylamino]carbonyl]4-methyl-3-oxo- 1 H- 1,4-benzodiazepine-2-acetate Methyl (S)-7-carboxy~-methyl-3-oxo-2,3,4,5-tetrahydro-lH-1,4-benzodiazepine-2-acetate (112.3 mg, 0.38 rnmol) was weighed into a 100 mL round bottomed flask. CH3CN was added, followed by HOBt ~ H~O (62.3 mg, 0.46 mmol~
and EDC (120.0 mg, 0.63 mmol). When the solids had all dissolved, diisopropylethylamine (0.1 mL, 0.57 mmol) was added, followed by a suspension of4,7-~lim~thoxy-2-(methylaminomethyl)be~-7i~ 7ole (96.8 mg, 0.44 mmol) in CH3CN cont5.inin~ diisopropylethylamine (0.1 mL, 0.57 mmol). After 2.5 days at RT, the reaction was evaporated under vacuum. The residue was evaporated once with toluene, then was chromatographed on silica gel (CHCl3, then 5%
MeOH/CHCl3) to afford the title compound (152.0 mg, 80.0%): TLC R~ (5%
MeOH/CHCl3) 0.35; MS (ES) m/e 496.2 (M+H)t; H NMR (250 MHz, CDC13) o 7.25 (m, 2H), 6.56 (s, 2H), 5.36 (d, lH), 3.91 (s, 6H), 3.70 (s,3H), 3.08 (s, 3H).
d) (S)-2,3,4,5-Tetrahydro-7-[[[(4,7-dimetho~ybel-7il..i~1~7~ 2-yl)methyl]methylarnino]carbonyl]-4-methyl-3-oxo- 1 H- 1,4-benzodiazepine-2-acetic acid Following the procedure of Example 45(e), methyl (S)-2,3,4,5-tetrahydro-7-t[[~4,7-dimethoxybçn7.imic~7.(-1-2-yl)methyl]methylamino]carbonyl]-4-methyl-3-oxo-lH-1,4-benzodiazepine-2-acetate was saponified to afford the title compound (110.0 mg, 74%): MS (ES) m/e 482.2 (M+H)~. Anal Calcd for C~,~H27N5O6 ~ 0.75 H2O: C, 58.23; H, 5.80; N, 14.15. Found: C, 58.26; H, 5.59; N, 13.90.
Fxample 47 ~l~dlion of (S)-2.3.4.5-tetrahydro-7-rrr(4-methylbenzimidazol-2-yllmethyllmethylaminolcarbonyll-4-methyl-3-oxo-lH-1.4-benzodiazepine-2-acetic 5 a~id a) 1,2-Diamino-3-methylbenzene Following the procedure of Example 45(a), except ~ub~ li..g 2-methyl-6-nitro~nilin.- (3.0204 g, l9.g8 mmol) for the 4-amino-3-nitrobenzotrifluoride, the title compound (2.4815 g) was prepared. This was used without char~( le, i7~1 ;on.
b) 2-[N-(Benzyloxycarbonyl)-N-methyl]aminomethyl-4-methylben7imitl:~7ole C bz-sarcosine (4.6466 g, 19.92 mmol) was dissolved in dry THF in a 100 mL
round-bottomed flask. Triethylamine (3.0 mL, 21.57 mmol) was added, followed by isobutylchlolufol-llate (2.8 mL, 21.59 mmol). The white reaction mixture was stirred at RT for Q.5 hr, then was added to a solution of 1,2-diamino-3-methylbenzene (2.4815 g) in dry THF at -20 to -30~C. After 20 min, the reaction was warmed to RT and stirred there for 16 hr. The reaction was evaporated under vacuum and the residue was partitioned between EtOAc and 1.0 N NaHCO3. The layers were separated, and the aqueous layer was extracted with EtOAc. The combined organics were dried (MgSO4), filtered, and evaporated under vacuum. The residue was reconcentrated from toluene and the dried solid was dissolved in glacial AcOH (150 mL). The solution was heated at 110~C for 18 hr, then was concentrated under high vacuum. The residue was reconcentrated from toluene, then was adsorbed onto silica gel and loaded onto a dry silica ge} flash chromatography column. The column was eluted with 1: 1 CH2Cl2/Et2O to afford the title compound (3.1586 g, 51%): MS (ES) m/e 310.2 (M+H)~; 'H NMR (250MHz, CDCl3) ~ 7.35-7.01 (m, 10H), 5.00 ~s, 2H), 4.72 (s, 2H), 2.99 (s, 3H), 2.55 (s, 3H).
c) 4-Methyl-2-(methylaminomethyl)ben7imi<1~7Ole 30 Following the procedure of Example 45(c), except subslil~li.. g 2-[N-(ben_yloxycarbonyl)-N-methyl]aminomethyl-4-methylben7imit1~37.01e for the 2-[N-(benzyloxycarbonyl)-N-methyl]aminomethyl-5-trifluoromethylb~on7imi(1~7nle, the title compound (2.9916 g, qn~ v~) was prepared: 'H NMR (250 MHz, CDCl3) o 7.36-7.01 (m, 4H), 4.01 (s, 2H), 2.52 (s, 3H), 2.41 (s, 3H).
d) Methyl (S)-2,3,4,5-tetrahydro-7-[[[(4-methylben7imi~1~7ol-2-, yl)methyl]methylamino]carbonyl]-4-methyl-3-oxo- 1 H- 1,4-benzodi~epine-2-acetate W O 97/24119 PCTrUS96120748 Methyl (S)-7-carboxy-4-methyl-3-oxo-2,3,4,5-tetrahydro-lH-1,4-benzodiazepine-2-acetate (188.5 mg, 0.64 mmol) was weighed into a 100 mL round bottomed flask. CH3CN was added, followed, sequentially, by HOBt ~ H2O (103.5 mg, 0.77 mmol), EDC (149.3 mg, 0.78 mmol), and diisopropylethylamine (0.15 ml, 0.86 S mmol). After 15 min, a solution of 4-methyl-2-(methylaminomethyl)ben7imi~1~7ole (273.8 mg, 1.56 mmol) in CH3CN was added. CH2Cl2 (5 mL) was added to dissolve some material. After 18 hr at RT, the reaction was concentrated, and the residue was chromatographed on silica gel (5% MeOH/CHCl3) to afford the title compound (307.3 mg, q~l~ntifz~tive): MS (ES) rn/e 450.2 (M+H)t; 'H NMR (250 MHz, CDCl3) ~ 7.23-7.03 (m, 7H), 6.41 (br s, lH), 5.33 (d, J = 16.3 Hz, lH), 3.69 (s, 3H), 3.46 (s, 3H), 3.10 (s, 3H).
e) (S)-2,3,4,5-Tetrahydro-7-[[L(4-methylbçn7imidz-~ol-2-yl)methyl]methylamino]carbonyl3-4-methyl-3-oxo- lH- 1,4-benzodiazepine-2-acetic acid Following the procedure of Example 45(e), methyl (S)-2,3,4,5-tetrahydro-7-[[[(4-methylbçn7imicls-7ol-2-yl)methyl]methylamino]carbonyl]-4-methyl-3-oxo-lH-1,4-benzodiazepine-2-acetate ~307.3 mg, 0.68 mmol) was saponified to afford the title compound (243.9 mg, 82%): MS (ES) rn/e 436.2 (M+H)+. Anal Calcd for C23H25N504 -2.75 H2O: C, 56.96; H, 6.34; N, 14.44: Found: C, 56.72; H, 6.27; N, 14.26.
F.x~m~G~le 48 ~dLion of (S)-2.3.4.5-tetrahydro-7-rrr(4-aza-5.7-dim~tllylbe~ idazol-2-yl)methyllme~llylaminolcarbonyll-4-methyl-3-oxo- 1 H- 1.4-benzodiazepine-2-acetic a) 2-Amino-4,6-dimethyl-3-nitropyridine 2-Arnino-4,6-dimelhyl~ylidine (5.55 g, 45.43 mmol) was weighed into a 500 mL round bottomed flask. The flask was cooled to -78~C. Concentrated HzS04 (25 mL, 450 mmol) was added, followed by concentrated HNO3 (3.5 mL, 56.0 mmol).
The ~ui~Lul~ became a solid frozen mass. The cooling bath was removed and the reaction was allowed to warm to RT. After about 15 min there was an exothermic reaction with the release of some nitrous oxide gas, and the reaction became a very dark red color. The reaction was heated at 85 - 90~C for 3 hr, then was cooled to RT, diluted with ice, and neutralized with 6 N NaOH (160 mL). The aqueous solution was extracted with EtOAc (3 x), and the combined EtOAc layers were dried .
W O 97/24119 PCTrJS96/20748 (MgSO4), filtered, and evaporated under vacuum. The resulting yellowish-orange solid was adsorbed onto silica gel and flash chromatographed on a dry silica gel colurnn. The column was eluted with 1: 1 CHCI3/Et2O to afford the title compound (1.0650 g, 14%): MS (ES) m/e 168.0 (M+H)'.
s b) 2,3-Diamino-4,6-dim~;lhyl~ylidine - Following the procedure of Example 45(a), except substituting 2-amino-4,6-dimethyl-3-nitl~ylidine (1.0650 g, 6.37 mmol) for the 4-amino-3-niLluben~otrifluoride, the title compound (836.1 mg, 95.7%) was prepared. This was used without char~ct~ri7~tion.
c) 4-Aza-2-[N-(benzyloxycarbonyl)-N-methyl]aminomethyl-5,7-dimethylbenzimidazole Following the procedure of Example 45(b), except sub~ 2,3-diamino-4,6-dhllt;lllylpyridine (836.1 mg, 6.09 mmol) for the 3,4-diaminobenzotrifluoride, the title compound (1.2273 g, 62%) following silica gel chromatography (3%
MeOH/CHCl3): MS (ES) m/e 325.0 (M~H)+.
d) 4-Aza-2-(methylaminomethyl)-5,7-dimethylben7imitl:~7ole Following the procedure of Example 45(c), except substituting 4-aza-2-[N-(benzyloxycarbonyl)-N-methyllarninomethyl-5,7-dimethylben7imi~1~7.ole (1.2273 g,3.78 mmol) for the 2-[N-(benzyloxycarbonyl)-N-methyl~aminomethyl-5-trifluoromethylbenzimidazole, the title compound was obtained as a white powder following trituration with Et2O. This material was used without characterization.
e) Methyl (S)-2,3,4,5-tetrahydro-7-[[[(4-aza-5,7-dimethylbe7imi~1~7c!1-2-yl)methyl]methylamino~carbonyl~-4-methyl-3-oxo- 1 H- 1,4-benzodiazepine-2-acetate Methyl (S)-7-carboxy-4-methyl-3-oxo-2,3,4,5-tetrahydro- lH- 1,4-benzodiazepine-2-acetate (175.0 mg, 0.60 mmol) was weighed into a 100 mL round bottomed flask. CH3CN (10 mL) was added, followed sequentially, by HOBt ~ H~O
(115.9 mg, 0.86 mmol), EDC (124.9 mg, 0.65 mmol), and diisopropylethylamine (0.13 mL, 0.75 mmol). A ~uspel sion of 4-aza-2-(methylaminomethyl)-5,7-dimethylben7imicl~7~-1e (144.5 mg, 0.76 mmol) and diisopropylethylamine (0.13 mL, 0.75 mmol) in CH3CN was added, and the reaction was stirred at RT. After 22 - 35 hr, the reaction was evaporated under V~l~;UUlll, and the residue was co-evaporated with toluene. Silica gel chromatography (3% MeOH/CHCl3 (1 L) then 5%
W O 97/24119 PCTrUS96120748 MeOH/CHCI3) gave the title compound (76.9 mg, 28%): MS (ES) m/e 465.2 (M+H)+.
f) (S)-2,3,4,5-Tetrahydro-7-[[[(4-aza-5,7-dimethylben7imi~ 7OI-2-yl)methyl]methylarnino]carbonyl-4-methyl-3-oxo-lH-1,4-benzodiazepine-2-acetic acid Methyl (S)-2,3,4,5-tetrahydro-7-[[[(4-a_a-5,7-dimethylbç7imi~7ol-2-yl)methyl]methylamino]carbonyl]-4-methyl-3-oxo- 1 H- 1,4-benzodiazepine-2-acetate (76.9 mg, 0.17 mmol) was dissolved in MeOH (5 mL) and HlO (5 rnL), and 1.0 N
- 10 NaOH (0.5 mL, 0.5 mmol) was added. After 24 hours at RT, the reaction was neutralized with 1.0 N HCl (0.5 mL), and the solvents were evaporated under vacuum. ODS chromatography (gradient: 5% CH3CN/H20 cont"inin~ 0.1% TFA
(500 mL), then 10% % CH3CN/H20 cont"ining 0.1 % TFA (500 mT~), then 15% %
CH3CN/H~0 cont"ining 0.1% TFA (500 mL), then 30% % CH3CN/EI~0 cont~ining 0.1 % TFA (500 mL)) gave a residue which was co-evaporated once with toluene anddried under high vacuum. The resulting residue was dissolved in MeOH (5 mL) and precipitated with Et,O. The white solid was collected on a sintered glass funnel and dried in a vacuum desiccator overnight to yield the title compound (52.0 mg, 68%):
HPLC (ODS column, 1.5 mL/rnin; gradient 5-50% CH3CN/H20 cont~ining 0.1%
TFA) tR 12.38 min; MS (ES) m/e 451.2 (M+H) ' . Anal Calcd for Cr,H~6N604 1 H20 1 CF3CO2H: C, 51.55; H, 5.02; N, 14.43: Found: C, 51.34; H, 5.00; N, 14.41.
Fx~ml?le 4g ~l~p~dtion of (S)-2.3.4.5-tetrahydro-7-rrr(5.6-difluoroben7imi~1~7ol-2-yl)methyllmethylaminolcarbonyll-4-methyl-3-oxo- 1 H- 1.4-benzodiazepine-2-açeticacid a) 1,2-Diarnino-4,5-difluorobenzene Following the procedure of Example 45(a), except substit-lting 4,5-difluoro-2-nitroaniline (2.0 g, 11.49 mmol) for the 4-amino-3-nitTobellzol~ ori~ , the title compound was prepared. This was used without characterization.
b) 2-[N-(Benzyl-~yca~bonyl)-N-methyl~aminomethyl-5,6-difluorobel-7i . . . i~ 7ole Following the procedure of Example 47(b), except substituting 1,2-~ mino-4,:~-difluoro~en~ne. for the 1,2~ mino-3-methylbenzene, and running the AcOH
cyclization step at 80~C instead of at 110~C, the title compound (1.3767 g, 36%) was W O 97/2411g PCTrUS96/20748 prepared: TLC Rf (1: 1 CH2C12/Et2O) 0.42; MS (ES) m/e 332.0 (M+H)+; IH
NMR (250 MHz, CDC13) ~ 7.50-7.14 (m, 8H), 5.13 (s, 2H), 4.61 (s, 2H), 3.06 (s, 3H).
S c) 5,6-Difluoro-2-(methylaminomethyl)ben7imid~7Ole Following the procedure of Example 46(b), except substituting 2-[N-- (benzyloxycarbonyl)-N-methyl]aminomethyl-5,6-difluorobçn7imi-1~7ole (1.3767 g, 4.16 mmol) for the 2-[N-(benzyloxycarbonyl)-N-methyl]~minom~thyl-4,7-~lim~thoxybe~ mdazole, the title compound (875.6 mg, qll~ntit~tive) was prepared:
10 MS (ES) m/e 198.0 (M+H)'.
d) Methyl-(S)-2,3,4,5-tetrahydro-7-[[[5,6-difluoroben7imill~7ol-2-yl)methyl]methylamino]carbonyl]-4-methyl-3-oxo- 1 H- 1,4-benzodiazepine-2-acetate Methyl (S)-7-carboxy-4-methyl-3-oxo-2,3,4,5-tetrahydro-lH-1,4-15 benzodiazepine-2-acetate (415.7 mg, 1.42 mmol) was taken up in CH3CN, and HOBt ~ H2O (209.3 mg, 1.55 mmol) and EDC (314.9 mg, 1.64 mmol) were added. After 5 min, and diisopropylethylamine (0.25 mL, 1.64 mmol) was added, which produced a clear, colorless solution. A solution of 5,6-difluoro-2-(methylaminomethyl)ben7imicl~701e (284.5 mg, 1.44 rnmol) in CH3CN was added.
20 After 30 minutes, the reaction became slightly turbid, so more diisopropylethylamine (0.25 mL) was added, which made the reaction again clear and colorless. After 24 hr, the reaction was evaporated under vacuum. The residue was co-evaporated once with toluene, then was chromatographed on silica gel (CHCl3 (0.25 L), then 2% MeOH/CHCl3 (1.5 L), then 5% MeOH/CHCl3) to afford 25 the title compound (456.8 mg, 68%): MS (ES) m/e 472.2 (M+H)t; 'H NMR (250 MHz, CDCl3) o 7.34-7.08 (m, 6H), 6.44 (br s, lH), 5.39 (d, J = 16.2 Hz, lH), 3.70 (s, 3H), 3.14 (s, 3H), 2.96 (s, 3H).
e) (S)-2,3,4,5-Tetrahydro-7-[[[(5,6-difluorobçr 7imi~ 7ol-2-30 yl)methyl]methylamino]carbonyl]-4-methyl-3-oxo- lH- 1,4-benzodiazepine-2-acetic acid Methyl (S)-2,3,4,5-tetrahydro-7-[[[5,6-difluoroben7imi~1~7-1-2-yl)methyl]methylamino]carbonyl] -4-methyl-3-oxo- l H- 1,4-ben70~ 7~pine-2-acetate (456.8 mg, 0.97 mmol) was dissolved in MeOH (10 mL) and H2O (10 ml). 1.0 N
35 NaOH (3.0 mL, 3.0 mmol) was added and the reaction was stirred at RT. After 18 hr, the reaction was neutralized with 1.0 N HCl (3.0 mT .). A white precipitate formed which was collected on a sintered glass filter and dried in a vacuum W O 97~4119 PCTrUS96/20748 desiccator. ODS chromatography (gradient: 10% CH3CN/H20 Cont~ining 0.1% TFA (500 rnL), then 18% % CH3CN/H20 cont~inin~ 0.1% TFA (500 mL), then 25% % CH3CN/H20 cont~inin~ 0.1% TFA (500 mL)) gave a residue which was co-evaporated once with toluene. The rçslllting residue was dissolved in a small 5 amount of MeOH and precipitated with Et2O to give the title co~ oulld (330.9 mg) as a white powder: HPLC (ODS column; 1.5 mL/min; gradient 5-50% CH3CN/H~O
contzlining 0.1% TFA) tR = 14.12 min; MS (ES) m/e 458.2 ~M+H)~. Anal Calcd for C22H2,N504F2 ~ 2.5 H2O: C, 52.57; H, 5.22; N, 13.94: Found: C, 52.76; H, 5.15; N, 67.
F.x~m~ple 50 Preparation of (S)-2.3.4.5-tetrahydro-7-rr~(4-:~7~-5-methylbe1l7i..lid~7.ol-2-yl)methylJamirlolcarbonyll-4-m~o-tllyl-3-oxo-lH-l~4-benzodiazepine-2-acetic acid a) Methyl (S)-2,3,4,5-tetrahydro-7-[[~(4-aza-5-methylben7imi-1~7nl-2-yl)methyl]amino]carbonylJ-4-methyl-3-oxo- lH- 1,4-benzodiazepine-2-acetate Methyl (S)-7-carboxy-4-methyl-3-oxo-2,3,4,5-tetrahydro- 1 H- 1,4- -benzodiazepine-2-acetate (228.8 mg, 0.78 mmol) was taken up in CH3CN, and HOBt ~ H~O (154.2 mg, 1.14 mmol), EDC (179.4 mg, 0.94 mmol), and diisopropylethylamine (0.50 ml~ ,, 0.94 rnmol) were added sequentially. A solution of 2-(aminomethyl)-4-aza-5-methylben7imi(1~7(1e dihydrochloride (125.4 mg, 0.77 mmol) in CH3CN /DMF was added, and the reaction was stirred at RT. After 24 hr, the reaction was evaporated under vacuum~ and the residue was co-evaporated oncewith toluene. Silica gel chromatography (CHCl3 (0.25 L) then 3% MeOH/CHCl3 (0.5 L), then 5% MeOH/CHCl3) gave the title compound (159.9 mg, 48%): MS (ES) m/e 437.2 (M+H)+.
b) (S)-2,3,4~5-Tetrahydro-7-[~[(4-aza-5-methylbenzimidazol-2-yl)methyl]amino]carbonyl]-4-methyl-3-oxo- lH- 1,4-benzodia_epine-2-acetic acid Following the procedure of Fx~mrle 48(f), methyl (S)-2,3,4,5-tetrahydro-7-[[[(4-aza-5-methyl~en7imi~ 7Ol-2-yl)methyl]aminoJcarbonyl]-4-methyl-3-oxo-lH-1,4-benzodiazepine-2-acetate (159.9 mg, 0.37 mmol) was saponified and purified to afford the title compound: MS (ES) m/e 423.39 (M+H)~. Anal. Calcd for C2,H22N6O4 - 0.5 H2O 1.25 TFA: C, 52.64; H, 5.02; N, 16.74. Found: C, 52.65; H, 5.02; N, 16.74.
Fx~m,ple 51 Preparation of (S)-2.3.4.5-tetrahydro-4-m~tllyl-7-rrr(4-nitrobçn7imicla7ol-2-yl)methyl~methylaminolcarbonyll-3-oxo-lH-1.4-benzo~ 7~pine-2-acetic acid a) 2-[N-(tert-Butoxycarbonyl)-N-methyl]aminomethyl-4-nitroben7imi<1~7ole e Boc-sarcosine (2.0320 g, 10.74 mmol) was dissolved in dry THF and cooled in a dry-ice/acetone bath to -15~C. Triethylamine (5.0 mL, 3.6375 mmol) was added, followed by isobutylchlor~fullllate (1.5 mL, 11.56 mmol). After 0.5 hr, the mixture was added to a solution of 1,2-diamino-3-nitrobenzene (1.3047 g, 10.77 mmol) in dry THF at -20~C, and the reaction was allowed to warm to RT. After 24 hr, the reaction was evaporated under vacuum and the residue was partitioned between EtOAc and 1.0 N NaHCO3. The layers were separated, and the aqueous layer was extracted with EtOAc. The combined organics were dried (MgSO4), filtered, and evaporated under vacuum. The residue was dissolved in glacial AcOH(100 mL), and the solution was heated at 75~C. After 24 hr, the reaction was evaporated under vacuum, and the residue was co-evaporated with toluene (2 x).
The m~t~ri~l was adsorbed onto silica gel and flash chromatographed on a dry silica gel column (gradient: CHC13 (0.5 L), then 2% MeOH/CHCl3 (1 L), then 5%
MeOH/CHCl3) to afford the title compound (2.2089 g, 75%): 'H NMR (250 MHz, CDCl3) ~ 8.10 (dd, 2H), 7.40-7.32 (m, lH), 4.69 (s, 2H), 3.02 (s, 3H), 1.54 ~s, 9H).
b) 2-(Methylaminomethyl)-4-niliuben,i.l.i(1~7Ole 2-[N-(tert-Butoxycarbonyl)-N-methyl3aminomethyl-4-nitroben7.imi~ 7.ole (2.2089 g, 8.05 mmol) was treated with 4 N HCl in dioxane at RT. After the addition, there was an imm~ te precipitation of a white solid. After 4 hr, the reaction was evaporated under vacuum and the residue was triturated with diethylether to give the title compound (1.639 g) as a white solid. This was used without chara~ ation.
c) Methyl-(S)-2,3,4,5-tetrahydro-7-1[[(4-nitrobe~7.imi~1~7.ol-2-yl)methyl]methylamino~carbonyl-4-methyl-3-oxo- 1 H- 1,4-benzodiazepine-2-acetateFollowing the procedure of Example 49(d), except substitllting 2-(methylaminomethyl)4-nitroben7.imi~1~7c-1e for 5,6-difluoro-2-(methylaminomethyl)ben7imi-1~7ole, the title compound (292.9 mg, qn~ntit:~tive) was plep~d: MS(ES) m/e 481.2 (M~H)+.
CA 0224l633 l998-06-26 W O 97/24119 PCTrUS96/20748 d) (S)-2,3,4,5-Tetrahydro-7-t[[(4-nitro~çn7imitl~7ol-2-yl)methyl]methylamino]carbonyl-4-methyl-3-oxo- l H- l ,~bP,n7.o~ .pine-2-acetic acid Following the procedure of Example 45(e), methyl ~S)-2,3,4,5-tetrahydro-7-[[[(4-nitrobenzimidazol-2-yl)methyl]methylamino]carbonyl-4-methyl-3-oxo- 1 H- 1,4-benzodiazepine-2-acetate (292.9 mg, 0.61 mmol) was saponified to afford the title compound (211.0 mg, 68%): MS (ES) m/e 467.4 (M+H)+. Anal. Calcd for C22H22N6O6 2.5 H20: C, 52.12; H, 5.27; N, 16.58: Found: C, 52.07; H, 4.97; N, 16.40.
F.xzln~?le 52 .,.tion of (S)-2.3.4.5-tetrahydro-7-rrr(4-arninoben7imi-l~7ol-2-yl)m~thyllmethylaminolcarbonyl-4-methyl-3-oxo-lH- 1 ~4-benzodiazepine-2-acetic ~i~
a) (s)-2~3~4~5-tetrahydro-7-[[[(4-amino~rn7imi~ 7ol-2-yl)methyl] methylamino}carbonyl-4-methyl-3 -oxo- 1 H- 1,4-benzodiazepine-2-acetic acid (S)-2,3,4,5-Tetrahydro-7-[[[(4-nitroben7imir1~ol-2-yl)methyl]methylamino]carbonyl-4-methyl-3-oxo- 1 H- 1,4-benzodiazepine-2-acetic acid (108.7 mg, 0.21 mmol) was dissolved in MeOH, and a catalytic amount of 10%
PdlC was added. The reaction was purged with Hz, then was stirred at RT under H2(balloon). After 20 hr, the catalyst was removed by filtration through celite(g), and the filtrate was evaporated under vacuum. The resulting solid was dissolved in MeOH, reprecipitated with Et20, dried in a vacuum rlecicc;~tor, and purified by ODS
chromatography (gradient: H2O cont~inin~ 0.1% TFA (500 mL), then 5%
CH3CN/H20 cont~ining 0.1% TFA (500 mL), then 10% % CH3CN/H70 Cont~inin~
0.1% TFA (500 mL), then 15% % CH3CN/H20 confz-inin~ 0.1% TFA (500 rnL), then 20% % CH3CN/H20 cont~inin~ 0.1% TFA (500 mL), then 25% % CE~3CN/Hz0 contz~ining 0.1% TFA (500 rnL), then 30% % CH3CNIH20 cont:~lininp: 0.1% TFA
(500 mL)). The rçsnltin~ m~tPri~l was co-evaporated once with toluene then was triturated with diethyl ether to afford the title compound (34.7 mg): MS (ES) m/e 437.5 (M+H)~; Anal. Calcd for C~24N6O4 1.5 H2O 1.5 TFA: C, 47.32; H, 4.53; N.
13.24: Found: C, 47.35, H, 4.86; N. 13.61.
Fx~m;l?le 53 W O 97/24119 PCTnUS96120748 ?ari.lion of 23.4~5-tetrahydro-7-rrr(lR~-fb~n7imi(1~7ol-2-yl)ethyllmethylarninolcarbonyll-4-m~thyl-3-oxo-lH-1.4-benzodia_epine-(2S~-aceticacid -a) 2-[1 (R)-~N-(Benzyloxycarbonyl)-N-methyl]aminoethyl~ben7.imifl~7.ole Following the procedure of Example 47(b), except ~ub~liluLillg Cbz-N-methyl-D-alanine for the Cbz-sarcosine, and substituting 1,2-phenylen~ mine for the 1,2-diamino-3-methylbenzene, and running the AcOH cyclization step at 80~C
10 instead of at 110~C, the title compound was prepared: MS (ES) m/e 310.2 (M+H)+.
b) 2-[1(R)-(Methylaminoethyl)]ben7.imi~ 7.01e Following the procedure of Example 46(b), except sub~ ulillg 2-[l(R)-tN-~benzyloxycarbonyl)-N-methyl]aminoethyl]ben7.imi(1~7.01e for the 2-[N-15 (benzyloxycarbonyl)-N-methyl]aminomethyl-4,7-dimethoxyben7imic~:~7.ole, the title compound (276.0 mg, 48%) was prepared: MS(ES) m/e 176.2 (M+H)+.
c) Methyl 2,3,4,5-tetrahydro-7-[[[(lR)-(ben7.imi~1~7.c!1-2-yl)ethyl]methylamino]ca~bollyl]-4-methyl-3-oxo- 1 H- 1,4-benzodiazepine-2-acetate Following the procedure of Example 49(d), except substituting 2-[l(R)-(methylaminoethyl)]benzimidazole for the 5,6-difluoro-2-(methylaminomethyl)1)~ll7.il"i(1~7.ole, the title compound (203.5 mg, 90%) was prepared: MS (ES) m/e 450.5 (M+H)~.
25 d) 2,3,4,5-Tetrahydro-7-[~[(lR)-(bçn7.imi-1~7.ol-2-yl)ethyl]methylamino]carbonyl]-4-methyl-3-oxo- 1 H- 1,4-benzodiazepine-2-acetic acid Following the procedure of Example 49(e), methyl 2,3,4,5-tetrahydro-7-[[[(lR)-(b~7imi~1~7.ol-2-yl)ethyl]methylamino~ca~bonyll-4-methyl-3-oxo-lH-1,4-benzodiazepine-2-acetate was saponified to afford the title compound (179.3 mg, 30 75%) following ODS chromatography: MS (ES) m/e 436 5 (M~H)+. Anal. Calcd for C23H25N504 0.75 H20 0.75 TFA: C, 55.01; H, 5.14; N, 13.10; Found: C, 54.98;
H, 5.42; N. 12.75.
F.x~n~le 54 CA 0224l633 l998-06-26 W O 97/24119 PCTrUS96/20748 Pl~alion of ethyl fS)-2.3~4.$-tetrahydro-7-rrr(4-Aza-5-met~lylben7.imid~ol-2-yl)methyllaminolcarbonyll4-metlly1-3-oxo- 1 H- 1.4-'~n70~ 7~pine-2-acetate S a) Ethyl (S)-2,3,4,5-tetrahydro-7-[[[(4-Aza-5-methylben7imi~1~701-2-yl)methyl]amino]carbonyl]4-methyl-3-oxo- lH- 1 .4-bt~n70tli~7~pine-2-acetate (S)-2,3,4,5-Tetrahydro-7-[[[(4-aza-S-methylbçn7.imi~7.oI-2-yl)methyl]arnino]carbonyl]-4-methyl-3-oxo- 1 H- 1 ,4-benzodiazepine-2-acetic acid ~0.5 g) was dissolved in EtOH, and the solution was cooled in an }ce bath to 0~C.
Gaseous HCI was bubbled into the solution until the solution was saturated, then the flask was sealed with a rubber septum and the cooling bath was removed. The reaction was stirred at RT for 20 hr, then the solvents were evaporated under vacuum. The residue was co-evaporated three times with toluene (3 x), then was dissolved in EtOH and precipitated with Et20. The solid was collected on a sintered glass funnel and dried in a vacuum ~iesiccatQr overnight to afford the title compound (483.9 mg): MS (ES) m/e 451.4 (M+H)~. Anal. Calcd for C23H26N6O, ~ HCl ~ 1.375 H20: C, 53.98; H, 5.86; N, 16.42. Found: C, 54.00; H, 5.82; N, 16.42.
F.xz-m~?le S5 PIG~al~Lion of 2.3.4.5-tetrahydro-7-rrr(lS)-(ben7imi-1~7.ol-2-yl)etl~yllmethyl~minolcarbonyll-4-methyl-3-oxo-lH-1 .4-benzodi~epine-~2S)-aceticacid a) 2-[l(S)-[N-(tert-Butoxycarbonyl~-N-methyl~aminoethyl]ben7imi~1~701e Following the procedure of Example 51(a) except substituting Boc-N-methyl-L-alanine for the Boc-sarcosine, and sub..liLuLillg 1,2-phenylene~ min~ for the 1,2-diamino-3-nitroben7~n~, the title compound (1.7792 g, 65%) was prepared following recryet:llli7~tion from CHCI3/hexanes: MS (ES) m/e 276.4 (M+H)~.
b) 2-[l(S)-(Methylaminoethyl)]ben7imi~1~7.ole Following the procedure of Example 51(b), except substituting 2-[l(S)-~N-(tert-butoxycal'L,ollyl)-N-methyl]aminoethyl]bel-7i . . .i-i~7Qle for the 2-[N-(tert-butoxycarbonyl)-N-methyl]aminomethyl4-nitroben7imi~ 701e, the title compound 35 was prepared. This was used without charactt-ri7~tion.
W O 97/24119 PCT~US96/20748 c) Methyl (S)-2,3,4,5-tetrahydro-7-[[t(1 S)-(ben7.imi~1~701-2-yl)ethyl]methylamino]carbonyl]-4-methyl-3-oxo- 1 H- 1,4-bçn7.o~ 7P-pine-2-acetate Following the procedure of Example 49(d), except substihlting 2-[l(S)-(methylaminoethyl)]ben7.imitl~7.(1e for the 5,6-difluoro-2-S (methylaminomethyl) ben7.imi~1~7.01e, the title compound (414.7 mg, 88%) was prepared: MS (ES) m/e 450.2 (M+H)+.
d) 2,3,4,5-Tetrahydro-7-t[[(lS)-(ben7.imi~ 7.~1-2-yl)ethyl]methylamino]carbonyl]-4-methyl-3-oxo- lH- 1,4-benzodiazepine-2-acetic acid 1(~ Following the procedure of Example 45(e), methyl (S)-2,3,4,5-tetrahydro-7-[[[(lS)-(ben7.imi~1~7.ol-2-yl)ethyl~methylamino]carbonyl]-4-methyl-3-oxo-lH-1,4-benzodiazepine-2-acetate was saponified to afford the title compound (117.2 mg):MS (ES) m/e 436.2 (M+H)+. Anal Calcd for C23H~5N504 0.75 H20 0.75 TFA: C, 55.05; H, 5.14; N, 13.10; Found: C, 55.14; H, 5.38; N, 13.04.
Ex~mple S6 P~ io~ of 2.3.4.5-tetrahydro-7-rrr(lS)-(b~n7imi~1~7.ol-2-yl)ethyllaminolcarbonyll-4-methyl-3-oxo-lH-1.4-ben7.o~;~7.~pine-(2S)-acetic acid a) 2-r 1 (S)-(tert-Butoxycarbonyl)aminoethyl]ben7imi~1~7.ole Following the procedure of Example 51 (a) except substituting Boc- L-alanine for the Boc-sarcosine, and sul)~LiLuLil~g 1,2-phenylene~ mine for the 1,2-diamino-3-nitroben7ene7 the title compound (714.7 mg, 25%) was prepared: MS
(ES) m/e 262.4 (M+H)+.
b) 2-[1 (S)-(Aminoethyl)]ben7imi(1~7.ole Following the procedure of Example 51(b), except subsliluli~lg 2-[l(S)-(tert-butoxycarbonyl)aminoethyl]ben7imi~1~701e for the 2-[N-(tert-butoxycarbonyl)-N-methyl]aminomethyl-4-nitroben7.imi~1~7.01e, the title compound was prepared. This was used without characterization.
c) Methyl 2,3,4,5-tetrahydro-7-[t[(1S)-(ben7imi~ 7.ol-2-yl)ethyl)]amino]carbonyl]-4-methyl-3 -oxo- l H- 1,4-ben 70~ 7epine-2-acetate Following the procedure of Example 49(d), except substit~ting 2-[l(S)-(aminoethyl)]ben7.imi~1~7.r 1e for 5,6-difluoro-2-(methylaminomethyl)be~- ~.i " ,icl~7.~1e, the title compound (270.7 mg, 80%) was p~ l.,d: MS (ES) m/e 436.0 (M+H~+.
W O 97/24119 PCT~US96120748 d) 2,3,4,5-Tetrahydro-7-t[[(lS)-(be~7imid~7ol-2-yl)ethyl)]amino]carbonyl]-4-methyl-3-oxo- lH-1,4-benzodiazepine-2-acetic acid Following the procedure of Example 45(e), methyl 2,3,4,5-tetrahydro-7-[[[(1 S)-(benzimidazol-2-yl)ethyl)]amino]carbonyl]-4-methyl-3-oxo- lH- 1,4-benzodia_epine-2-acetate was saponified to afford the title compound (158.1 mg, 61%): MS (ES) m/e 422.0 (M+H)+. Anal. Calcd for C2lH23N5O4 ~ 1.75 H20: C, 58.37; H, 5.90; N, 15.46; Found: C 58.17; H, 5.77; N, 15.08.
F.~z~ml?le 57 P~ lion of 2.3.4~5-tetr~l~ydro-7-rrr(lR~-(ben~ 7ol-2-yl)ethyllarninolcarbonyll-4-methyl-3-oxo-lH-1.4-benzodiazepine-(2S~-acetic acid a) 2-[1 (R)-(Benzyloxycarbonyl)aminoethyl]ben7imi~1~701e Following the procedure of Example 47(b~, except substituting Cbz-D-alanine for the Cbz-sarcosine, and substituting 1,2-phenylenedi~minc for the 1,2-diarnino-3-methylbenzene, and rurming the AcOH cyclization step at 80~C instead of at 110~C, the title compound (1.1455 g, 43%) was prèpared: MS (ES) m/e 296.4 (M+H) i .
b) 2-(l(R)-Aminoethyl)ben~imi-1~7O1e Following the procedure of Fx~mple 46(b), except sub~iLu~ g 2-[l(R)-(benzyloxycarbonyl)aminaethyl]ben7imi~ 701e for the 2-[N-(benzyloxycarbonyl)-N-methyl]aminomethyl-4,7-dimethoxyben~ 7Qle, the title compound (258.1 mg, 93%) was ~l~palt;d: MS(ES) m/e 161.9 (M+H)t.
c) Methyl 2,3,4,5-tetrahydro-7-[[[( lR)-(ben7imi~1~7~-1-2-yl)ethyl~amino]carbonyl]-4-methyl-3-oxo- 1 H- 1,4-benzodiazepine-(2S)-acetate Following the procedure of Example 49(d), except substituting 2-(l(R)-aminoethyl)ben7imi~701e for the 5,6-difluoro-2-(methylarninomethyl)benzimidazole, the title compound (263.6 mg, 84%) was prepared: MS (ES) m/e 436.3 (M+H)+.
d) 2,3,4,5-Tetrahydro-7-[[[(lR)-(be~7imi(i~7ol-2-yl)ethyl]amino]carbonyl]-4 methyl-3-oxo- lH- 1,4-ben7o~ 7topine-2-acetic acid W O 97/24119 PCTrUS96/20748 Following the procedure of F.xz-mple 49(e), methyl 2,3,4,5-tetrahydro-7-[[[(lR)-(b~n7imi~1~7ol-2-yl)ethyl]amino]carbonyl]~-methyl-3-oxo-lH-1,4-benzodiazepine-(2S)-acetate was saponified to afford the title compound (125.0 mg, 49%): MS (ES) m~e 422.0 (M+H)t. Anal. Calcd for C22H23N5O4 ~ 0.5 H20 ~ 1.25 HCl: C, 55.51; H, 5.35; N, 14.71. Found: C, 55.70; H, 5.47; N, 14.53.
- Example $8 Pr~a,dLion of (S)-2.3~4.5-tetrahydro-7-rrrCjmifl:~7o( 1 .2a)pyrid-2-yl)methyllmethylaminolcarbonyl~-4-methyl-3-oxo-lH-1.4-benzodiazepine-2-acetic acid a) 2-Carboethoxyirnidazo[ 1 ,2a]pyridine 2-Aminopyridine (4 g, 42.50 mmol) was dissolved in MeOH (50 mL). Ethyl bromopyruvate (8.3 g, 42.50 mmol) was added and the reaction was stirred at 70~Cfor 2 hr. The solvent was then elimin~te-l and the solution was neutralized with 1 M
NaOH. The reaction was extracted with EtOAc, and the combined EtOAc layers were washed with brine. Drying (MgSO4), filtration, concentration, and silica gel ilash column chromatography (2% MeOH/Cl2CH2) gave the title compound (4.5 g, 56%) as a pale yellow solid: 1H NMR (400 MHz, CDC13) o 1.43 (t, J = 7 Hz, 3H), 4.45(q,J=7Hz,2H),6.86(t,J=6.6Hz, lH),7.24(t,J=6.6Hz, lH).
b) 2-Hydroxymethylimicl~7O[1,2a]pyridine 2-Carboethoxyimidazo[1,2a]pyridine (0.5 g, 2.81 mmol) was dissolved in dry THF at 0~C, and then a solution of lithium ~ minl~m hydride (0.5 mL of a 1.0M in THF) was added. The reaction was allowed to warm to RT and stirred for 1 hr.
H2O (0.2 rnL) was added, followed by 15% NaOH (0.2 mL), and finally H2O (0.6 mL). The solids were removed by filtration and washed with hot THF (2 x 100 mL) and hot CHCl3 (4 x 100 mL). The filtrate and washings were combined and dried (MgSO4). Following filtration the solvents were removed under reduce pressure the residue was purified by silica gel flash column chromatography (5%
MeOH/Cl2CH2~ to obtain the title co~ u-ld (0.1 g, 25~o) as pale yellow liquid: 'H
NMR (400 MHz, CDCl3) o 4.85 (s, 2H), 6.76 (t, J = 6.8 Hz, lH), 7.15 (t, J = 6.8 Hz, lH), 7.53 (s, lH), 7.54 (d, J = 6.7 Hz, lH), 8.1 (d, J = 6.7 Hz, lH). MS (ES) m/e 149 (M + H) +.
,.
c) 2-Chloromethylimi~1~7o[ 1 ,2a]pyridine CA 0224l633 l998-06-26 W O 97/24119 PCTnjS96nO748 Thionyl chloride (0.4 mL, 3.2 mmol) was added to a solution of 2-hydroxymethylimi(l~70[1,2a]pyridine (0.4 g, 2.7 mmol) in CHC13 (30 mL) at 0~C
After stirring at RT for 1 hr, the suspension was poured into a mixture of ice, 10%
NaHCO3, and CHCl3. The layers were separated, and the aqueous phase was extracted with CHCI3. The organic extracts were combined and dried (MgS04).
Following filtration, the solvents were removed under reduce pressure to obtain the title compound (0.4 g, 89%). lH NMR (400 MHz, CDCl3) o 4.7 (s, 2H), 6.7 (t, lH~, -7.2 (t, lH), 7.5 (d, lH), 7.6 (s, lH), 8.0 (d, lH); MS (ES) m/e 167 (M + H)+.
d) 2-(Methylaminomethyl)imidazo[1,2a]pyridine Freshly condensed methylamine (15 mL) was added to a solution of 2-chloromethylimifl~70~1,2a]pyridine (317 mg, 2 mmol) in EtOH (5 mL) at 0~C, and the reaction mixture was allowed to stir at 0~C for 2 h. The solvent was then ~limin~tto~l and the residue was purified by reverse-phase column chromatography(C-18 silica gel, H20 cont~ining 0.1 % TFA). Lyophilization gave the title compound (461 mg, 87%) as a white solid: IH NMR (250 MHz, CDC13) o 2.5 (s, 3H), 4.0 (s, 2H), 6.7 (t, lH), 7.2 (t, lH), 7.5 (d, l~I), 7.6 (s, lH), 8.1 (d, lH). MS
(ES) m/e 162 (M + H)'.
- 20 e) Methyl (S)-2,3,4,5-tetrahydro-7-[[[(imidazo(1,2a)pyrid-2-yl)methyl]methylamino~carbonyl]-4-methyl-3-oxo- 1 H- 1,4-benzodiazepine-2-acetate Methyl (S)-7-carboxy4-methyl-3-oxo-2,3,4,5-tetrahydro- lH- I ,~
benzodiazepine-2-acetate (828.2 mg, 2.83 mmol) was taken up in CH3CN, and HOBt ~ H20 (398.1 mg, 2.92 mmol), EDC (562.9 mg, 2.94 mmol), and diisopropylethylamine (1 mL, 5.74 mmol) were added sequentially. When the solids had dissolved, a solution of 2-(methylaminomethyl)imi~ 70[1,2a]pyridine (460.7 mg, 2.86 mmol) and diisopropylethylamine (1.5 mL, 8.61 mmol) in CH3CN
was added, and the reaction was stirred at RT. After 24 hr, the reaction was concentrated, and the residue was co-~v~olat~d with toluene (2 x). The rçsl-lting residue was chromatographed on silica gel (5% MeOH/CHCl3) to give the title compound (694.6 mg, 56%): MS (ES) m/e 436.4 (M~H)~.
f) (S)-2,3,4,5-Tetrahydro-7-[[[(imi-1~7.o(1,2a)pyrid-2-yl)methyl]methylamino~carbonyl]-4-methyl-3-oxo- 1 H- 1,4-benzodiazepine-2-aceticacid Methyl (S)-2,3,4,5-tetrahydro-7-~[[(imidazo(1,2a)pyrid-2-yl)methyl]methylamino]carbonyl]-4-methyl-3-oxo- lH- 1,4-benzodiazepine-2-acetate 182 "
CA 0224l633 l998-06-26 W O 97/24119 PCT~US96/20748 (663.4 mg, 1.52 mmol) was dissolved in MeOH (10 mL). H2O (10 mL) was added, followed by 1.0 N NaOH (5 mL, 5.0 mmol), and the reaction was stirredat RT. After 20 hr, the reaction was neutralized with 1.0 N HCl (5 mL), and the solution was evaporated under vacuum. The residue was purified by ODS
chromatography (gradient: H2O containing 0.1% TFA (500 mL), then 5%
CH3CN/H20 con~aining 0.1% TFA (500 mL), then 10% CH3CN/H20 co~
- 0.1% TFA (500 mL), then 15% CH3CN/H2O cont~ining 0.1% TFA (500 mL), then 20% CH3CN/H2O cont~ining 0.1% TFA (500 rnL)) followed by rechromatography on ODS (gradient: H20 cont~inin~ 0.1~TFA (250 mL), then 10% CH3CN/H2O
cont~ining 0.1% TFA (1.5 L), then 20% CH3CN/H20 c~-nt~ininp 0.1% TFA (1 L).
Fractions containing pure m~t~ri~l were combined and concentrated. The residue was co-evaporated with toluene, then was dissolved in MeOH and reprecipitated with l~t20 to afford the title compound (96.4 mg): MS (ES) m/e 421.9 (M+H)~.
Anal. Calcd for C22H23N5O4 ~ 0.25 H2O ~ TFA: C, 53.38; H, 4.57; N, 12.97. Found:C, 53.68; H, 4.97; N, 12.94.
E~xample 59 Preparation of (+)-7-rrr(4~5-Dimethyl-lH-;mi~ ol-2-yl)methyllmethylaminolcarbonyll-2~3.4.5-tetrahydro-4-methyl-3-oxo-lH-1~4-benzodiazepine-2-acetic acid a) N-(Carbobenzyloxy)-N-(methyl)acetonitrile Cbz chloride (7.40 rnL, 49.3 mmol) was added slowly at RT to a solution of N methylaminoacetonitrile hydrochloride (5.0 g, 46.92 mmol) and triethylamine (13.4 mL, 96.2 mmol) in dichloromethane (200 mL). The reaction was stirred at RT18 h, and the mixture was washed with lN HCl, water and brine. The organic layerwas dried (MgSO4~ and concentrated to yield the title compound (6.97 g, 73%) as a clear oil.
b) N-(Carbobenzyloxy)-N-(methyl)aminothio~ret~mi~le Hydrogen sulfide was bubbled through a solution of N-(carbobenzyloxy)-N-(methyl)acetonitrile (15 g, 73.5 mmol) and triethylarnine (30.75 mL, 220.6 mmol) in DMF (250 mL). After 20 min, the flask was closed and the reaction was stirred atRT for 18 h. The reaction was then poured into 2 N NaHCO3 (1 L ) and extracted with dichloromPth~ne. The combined organic phase was washed with 1: 1 .
W O 97/24119 PCT~US96t20748 water/brine (5 x), dried (MgSO4), and concentrated to give a yellow oil which was purified by silica gel llash chromatography (step gradient, 40-50% ethyl acetate/hexane) to yield the title compound (12.26 g, 70%) as a white solid: MS
(ES) rn/e 239.0 [M+H]+.
s c) N-(Carbobenzyloxy)-N-methyl-S-(methyl)acetothioimidate Iodomt-.th~nP (17.66 mL, 283.6 mmol) was added at RT to a solution of N-(carbobenzyloxy)-N-(methyl)aminothio~et:lmide(6.75 g, 28.36 mmol) in acetone 100 mL). The solution was stirred at RT in the dark for 3 h, and the reslllting precipitate was filtered to yield the title compound (9.63 g, 89%) as a white solid:
MS (ES) m~e 253.4 [M+H]+.
d) (+)-2-Amino-3,3-dimethoxybutane Sodium cyanoborohydride was added to a solution of 3,3-dimethoxy-2-butanone (1.32 g, 10 mmol) and ammonium acetate (1.1 g, 100 mmol) in methanol (30 rnL). The pH was adjusted to 6 with methanolic HCl, and the reaction was stirred at RT overnight and concentrated. The residue was dissolved in water, and the pH was adjusted to 5 with aqueous HCl. The res-lltin~ solution was extractedwit_ ether (3 x), and the aqueous phase was basified to pH 10 with Na2CO3, and extracted with ether. The organic layer was dried (MgSO4) and concentrated to yield the title compound (1.1 g, 83%) as a clear oil: MS (ES) m/el34.2 [M+Hl+.
e) N-(Carbobenzyloxy)-N-methyl-(4,5-dimethyl- lH-imidazol-2-yl)meth~n~rnine A solution of (~)-2-amino-3,3-~l;m~tho~y~uLalle (1.05 g, 7.89 mmol) and N-(carbobenzyloxy)-N-methyl-S-(methyl)~< etofhi~ imidate (2.0 g, 5.26 mmol) in methanol (30 mL) was warmed at 60~C for 2 h, and concentrated to give a yellow oil. The crude oil was dissolved in 6 N HCl (30 mL) and stirred at RT 1 h. The solution was basified to pH 12 wieh aqueous NaOH and then extracted with dichlor~)m~th~ne. The combined organic phase was dried (MgSO4) and concentrated to give a brown oil which was purified by silica gel flash chromatography (4% methanol/dichloromethane) to yield the title compound (0.590 g, 41%) as a clear oil: MS (ES) m/e 274.0 [M+H~+.
f~ N-Methyl-(4,5-dimethyl-lH-imi~ 7,t~1-2-yl)~ nzlrninr.
A solution of N-(carbobenzyloxy)-N-methyl-(4,5-dimethyl-lH-imidazol-2-yl)mto.thz-nz~mine (0.35 g, 1.28 mmol) in methanol (15 mL) and glacial acetic acid (5 CA 0224l633 l998-06-26 W O 97/24119 PCTrUS96/20748 mL), cont~ining 10% Pd/C (0.035 g), was shaken in a H2 atmosphere (45 Psi) for 6 h. The reaction was filtered and filtrate conce~ dl~;d to give the title compound (0.22 g, 86%) as a brown oil which was used in the next step without further purification: MS (ES) m/e 140 [M+H]+.
g) Methyl (_)-7-[[[(4,5-dimethyl-lH-imi-1~7ol-2-yl)methyl]methylamino]carbonyl]-- 2,3,4,5-tetrahydro-4-methyl-3-oxo- 1 H- 1,4-benzodiazepine-2-acetate A solution of N-methyl-(4,5-dimethyl-lH-imi(1~7ol-2-yl)meth~n~minP (0.20 g, 1.1 mmol) and methyl (_)-7-carboxy-2,3,4,5-tetrahydro-4-methyl-3-oxo-1~-1,4-benzodia_epine-2-acetate (0.320 g, 1.1 mmol) in the presence of DIEA (0.287 ml, 1.65 mmol) was stirred at RT. EDC (0.316 g, 1.65 mmol) was then added, followed by DMAP (0.013 g, 0.11 mmol). The mixture was stirred at RT for 18 h, and concentrated to give an oil which was purified by silica gel flash chromatography (step gradient, 0.5-2% methanol/dichloromPthzlne) to yield the title compound (0.060 g, 9%) as a clear oil: MS (ES) m/e 414.2 [M+H]+.
h) (+)-7-[[[(4,5-Dimethyl-lH-imidazol-2-yl)methyl]methylamino]carbonyl]-2,3,4,5-tetrahydro-4-methyl-3-oxo- lH- 1,4-benzodiazepine-2-acetic acid 1 N Sodium hydroxide (3 eq) was added to a solution of methyl (+)-7-[[[(4,5-dimethyl-lH-imidazol-2-yl)methyl]methylamino]carbonyl]-2,3,4,5-tetrahydro-4-methyl-3-oxo-lH-1,4-benzodiazepine-2-acetate (0.060g, 0.145 mmol), the mixture was stirred at RT for 5 h, and concentrated. The residue was dissolved in water and the pH was adjusted to S with 50% acetic acid. The solution was concentrated andpurified by MPLC (ODS-AQ, 10% acetonitrile/water cont~inin~ 0.1% TFA, UV
detection at 220 nm) to yield the title compound (0.050 g, 86%) as a white solid:
MS (ES) m/e 400.2 [M+H]+.
Example 60 Preparation of 3-rr3-r2-(ben7imill~7- 1-2-yl)ethyllisoxazolin-SfR.S)-yllacetyllamino-3(R.S)-meth~ upalloic acid a) 4-(Ben7imi-1~7Ol-2-yl)- 1 -butene According to the general procedures of Preparation 4 in P50256-1, except ~ul~Li~ulillg 4-pentenoic acid for the Boc-sarcosine, the title compound is prepared.
-b) 4-( l -Toluenesulfonylben7imi~l~7Ql-2-yl~- l -butene W O 97124119 PCTrUS96/20748 Sodium hydride is added carefully to a solution of 4-(bP.n7.imi(1~7.01-2-yl)-1-butene (50 mmole) and 4-toluenesulfonyl chloride (55 mmole) in dry THF
(2~)0 mr .). The reaction is stirred at RT until complete, then is quenched withsaturated NH,,CI (200 mL~, and the mixture is extracted with EtOAc. The combined5 organic extracts are dried (MgSO4) and concentrated, and the residue is purified by silica gel chromatography to give the title compound.
-c) 4-(l-Toluenesulfonylben7.imi-1~7.ol-2-yl)-l-butanal Ozone is bubbled into a solutiolI of 4-(1-toluenesulfonylben7imi~l~7.Ql-2-yl)-l-butene (40 mmole) in CH2Cl~ (160 mL) and MeOH ~40 mL) at -78~C until the blue color persists, then the excess ozone is removed by bubbling argon through the solution. Dry dimethylsulfide (excess) is added, and the reaction is warmed to RT.
The reaction is stirred at RT until complete, then is concentrated, and the residue is chromatographed on silica gel to afford the title c~ oulld.
d) 4-(l-Toluenesulfonylben7.imi~1~7.(-1-2-yl)-l-butanal oxime Hydroxylamine hydrochloride (33 rnmole) is added to a solution of 4-(1-toluenesulfonylben7imi~1~7.ol-2-yl)-1-butanal (30 mmole) and anhydrous sodium acetate ~66 mmole) in MeOH (150 mL) at 0~C. The reaction is stirred at 0~C until20 complete, then is concentrated, and the residue is partitioned between H20 and EtOAc. The layers are separated, and the aqueous layer is extracted with EtOAc.
The combined organic layers are washed sequentially with 5% NaHCO3 and saturated brine, dried (MgSO4~, and concentrated to afford the title compound.
2~ e) 4-(l-Toluenesulfonylbenzimi~ 7.ol-2-yl)-l-butanoxirninoyl chloride According to the procedure of Example l(b) in WO 95/14682, except substituting 4-(1-toluenesulfonylbe~7.imifl~7.01-2-yl)-l-butanal oxime for the 4-cyanobenzoxime, the title compound is prepared.
f) tert-Butyl ~3-[2-(1 -toll-~neslllfonylbPn7imi(1~7ol-2-yl)ethyl]isoxazolin-5(R~S)-yl~acetate According to the procedure of Example 1 (d) of WO 95/14682, except sub.~ uling 4-(1-toluenesu~nylben7.imi<~ Ql-2-yl)-1-butanoximinoyl chloride for the 4-cyanobenzoximinoy~ chloride, and ~ub~liluling tert-butyl 3-butenoate for the methyl 3-butenoate, the title compound is ~l~al~d.
W O 97/24119 PCT~US96/20748 g) [3-[2-(1-Tnllleneslllfonylbenzimidazol-2-yl)ethyl]isoxazolin-S(R,S)-yl]acetic acid 4 M HCl in dioxane (10 mL) is added to a solution of tert-butyl [3-[2-(1-toluenesulfonylben7imi(1~7nl-2-yl)ethyl]isoxazolin-5~R,S)-yl]acetate (5 mmole) in C~I2CH2 (40 mL) at 0~C. The reaction is stirred at RT until complete, then is concentrated to afford the title compound.
h) Ethyl 3-[[3-[2-(1-toluenesulfonylben7imi-1~7ol-2-yl)ethyl]isoxazolin-5(R,S)-yl]acetyl]amino-3(R,S)-methylpropanoate EDC (1.2 mmole) is added to a solution of [3-[2-(1-toluenesulfonylbenzimidazol-2-yl)ethyl]isoxazolin-5(R,S)-yl]acetic acid (1 mmole), ethyl 3(R,S)-aminobutyrate (1.2 mmole), HOBt H2O (1.2 mmole), and diisopropylethylamine (4 mmole) in anhydrous CH3CN (5 rnL) at RT. The reaction is stirred at RT until complete, then is concentrated, and the residue is purified by silica gel chromatography to afford the title compound.
i) 3-[[3-[2-(Ben7imi-1~7ol-2-yl)ethyl]isoxazolin-5(R,S)-yl]acetyl]amino-3(R,S)-methylpropanoic acid 1.0 N LiOH (2.5 mmole) is added to a solution of ethyl 3-[3-[2-(1-toluenesulfonylb~ n7imi~1~7ol-2-yl)ethyl]isoxazolin-5(R,S)-yl]acetyl]amino-3(R,S)-methylpropanoate (0.5 mmole) in THF (2.5 ml). The reaction is stirred at RT until complete, then is neutralized with l.0 N HCl. The solution is concentrated and the residue is purified by reverse-phase chromatography to afford the title compound.
Fxample 61 Preparation of 3-{3.4-dihydro-8-rrr(ben7imi(1~7nl-2-yl)methyl~methy}aminolcarbonyll-1-methyl-2~5-dioxo-lH-1.4-benzodiazepine}-4-propanoic acid a) 2-Amino-4-iodobenzoic acid The title compound is prepared from the oxidation of 4-iodo-2-nitrotoluene to give 4-iodo-2-niLI~bell~oic acid according to the method of Sasson, et al., J. Org.
Chem. 1986, 51, 2880-2883, followed by reduction of the nitro group using iron and acetic acid.
W O 97/24119 PCTrUS96/20748 b) 4-Iodoisatoic anhydride To a mechanically stirred ice-cold solution of 2-amino-4-iodobenzoic acid (26.3 g, 0.1 mol), sodium carbonate ~10.6 g, 0.1 mol) and water (250 mL), is slowly added, via an addition funnel, a solution of phosgene in toluene (80 mL of 1.93 M
S solution). After 2 h, the precipitated product is i.~ol~te~l by filtration and the solid is washed cuccessively with water (200 mL), a 1: 1 mixture of ethanol/ether (300 mL),and ether (200 mL). Drying under vacuum yields the title compound.
c) N-(2-Amino-4-iodobenzoyl)-,13-alanine benzyl ester A magnetically stirred solution of 4-iodoisatoic anhydride (5.0 g, 0.0173 mol)"1~ alanine benzyl ester tosylate (5.85 g, 0.0173 mol), and dimethylaminopyridine (0.5 g, 0.0041 mol) in pyridine (35 rnL) is heated for 2 h at 80~C. The reaction mixture is allowed to cool to RT and concentrated in vacuo.
The resulting residue is dissolved in ethyl acetate (100 mL), and washed successively with 10% cupric sulfate (2 x 50 rnL), saturated sodium bicarbonate (I x 50 mL) and brine (1 x 50 mL). Drying (Na2SO4), filtration, concentration, and silica gel chromatography (1:1 EtOAc/hex~n~ s) gives the title compound.
d) N-(4-Iodo-2-methylaminobenzoyl)-,13-alar~ine benzyl ester A m~gnç~ lly stirred solution of N-(2-amino-4-iodobenzoyl)-13-alanine benzyl ester (2.0 mmol), 2,6-lutidine (0.35 mL, 3.0 mmol) and methyl iodide (0.19 mL, 3.0 mmol) in DMF (15 mL) is heated at 50~C for 15 h. The reaction mixture isallowed to cool to RT and concentrated in vacuo. The r~s-llting residue is dissolved in ethyl acetate (75 rnL), and washed succl ccively with 10% citric acid (1 x 50 mL), saturated sodium bicarbonate (1 x 50 mL) and brine (I x 50 mL). Drying (Na2SO4),filtration, concentration, and silica gel chromatography (gradient 35-65 %
EtOAc/hexanes) gives the title compound.
e) Benzyl 3-~3,4-dihyro-8-iodo- 1-methyl-2,5-dioxo- lH- 1,4-benzodiazepinel-4-propanoate To a cold (-30~C) m~gn~ti~:~lly stirred solution of N-(4-iodo-2-methylarninobenzoyl)-,13-alanine benzyl ester (0.305 g, 0.69 mrnol) and triethylamine (0.144 g, 1.04 mmol) in methylene chloride (3 mL) is added slowly a solution of a-bromoacetyl bromide (0.09 mL, 1.04 mmol) in methylene chloride (2 mL) under argon. The reaction mixture is allowed to warm to RT and stir for 2 h. The llli~lulG
is diluted with methylene chloride (40 ml ) and washed s-lcce~ivGly with 10% citric acid (1 x 50 rnL) and saturated sodium bicar~onate (1 x 50 mL), dried (Na2SO4), W O 97/24119 PCT~US96/20748 filtered and concentrated in vacuo. The resultin~ residue is dissolved in DMF (3 ml) and added via an addition funnel to a slurry of sodium hydride (25 mg, 1.04 mrnol) in DMF (2 mL) which is cooled to 0~C. After 2 h of stirring, the mixture is poured over an ice cooled solution of 10% citric acid (50 mL) and 5 extracted with ethyl acetate (3 x 40 ml). The combined extracts are washed with saturated sodium bicarbonate (1 x 50 mL), dried (Na2SO4), filtered and concentrated.
Silica gel chromatography (gradient 40-70% EtOAc/hexanes) gives the title compound.
f) Benzyl-3-[3,4-dihyro-8-t[[(ben7imi(1~7Ol-2-yl)methyl]methylamino]carbonyl]
methyl-2,5-dioxo- 1 H- 1,4-benzodiazepine]-4-propanoate A mixture of benzyl 3-[3,4-dihyro-8-iodo- 1-methyl-2,5-dioxo- lH-1,4-benzodiazepine]-4-propanoate(2 mmol), 2-(methylaminomethyl)ben~ill,idazole dihydrochloride (3 mmol), DIEA (1.8 mL, 10 mmol), and (Ph3P)2PdC12 (140 mg, 0.2 mrnol) in N-methyl-2-pyrrolidinone (20 mL) is heated to 110~C under CO
balloon for 3 h. The mixture is then concentrated and the residue is purified bysilica gel flash chromatography to give the title compound.
g) 3-[3,4-Dihydro-8-[[[(ber-7imi~1~7Ol-2-yl)methyl]methylamino]ca.bollyl]-l-methyl-2,5-dioxo- 1 H- 1,4-benzodiazepine]-4-propanoic acid A mixture of benzyl-3-[3,4-dihyro-8-[[[(bel,,.i...ifl~7Ql-2-yl)methyl]methylamino]carbonyl3 - 1 -methyl-2,5-dioxo- 1 H- 1,4-benzodiazepine3-4-propanoate (2 mmol) and 10% Pd/C (0.02 g) in ethanol (100 mL) is hydrogenated inan atmosphere of H2 (50 psi3 for 6 h. The catalyst is removed by filtration, and the filtrate is concentrated under vacuo to afford the title compound.
Fx~m~le 62 ~re~aldlion of 3-{4H-imi-l~7Q~1~2-al~1.41benzQdiazepine-5(6H)-l-methyl-6-oxQ-9-rrr(ber~7imi~ 701-2-yl)methyllmethylaminolcarbonyl~ }-4-propanoic acid a) Ethyl N-(2-amino-4-iodobenzoyl)-,B-alanine A m~gneti~lly stirred solution of 4-iodoisatoic anhydride (0.0173 mol)"l3 alanine ethyl ester hydrochloride (0.0173 mol), and dimethylaminopyridine (0.5 g, - 35 0.0041 mol) in pyridine (35 mL) is heated for 2 h at 80~C. The reaction mixture is allowed to cool to RT and concentrated in vacuo. The resulting residue is dissolved in ethyl acetate (100 mL), and washed successively with 10% cupric sulfate (2 x 50 W O 97/24119 PCT~US96/2~748 mL), saturated sodium bicarbonate (1 x 50 rr~ ) and brine (1 x 50 rnL).
Drying (Na2SO4), filtration, concentration, and silica gel chromatography (1:1 EtOAc/hexanes) gives the title compound.
b) Ethyl-3-[3 ,4-dihyro-8-iodo-2,5-dioxo- 1 H- 1 ,4-benzodiazepine]-4-propanoateTo a cold (-30~C) m:~gnPti~z~lly stirred solution of ethyl N-(2-amino-4-iodobenzoyl)-,B-alanine (0.69 mmol), and triethylamine (0.144 g, 1.04 mmol) in methylene chloride (3 rnL) is added slowly a solution of a~-bromoacetyl bromide (0.09 mL, 1.04 mmol) in methylene chloride (2 mL) under argon. The reaction mixture is allowed to warm to RT and stir for 2 h. The mixture is diluted with methylene chloride (40 mL) and wash snccç~ively with 10% citric acid (1 x 50 mT.) and saturated sodium bicarbonate (1 x 50 mL), dried (Na2SO;), filtered, and concentrated in vacuo. The rçsllltin~ residue is dissolved in DMF (3 mL) and added via an addition funnel to a slurry of sodium hydride (25 mg, 1.04 mmol) in DMF (2 mL) which is cooled to 0~C. After 2 h of stirring, the rnixture is poured over an ice cooled solution of 10% citric acid (50 mL) and extracted with ethyl acetate (3 x 40 rnL). The combined extracts are washed with saturated sodium bicarbonate ( 1 x 50 rnL), dried (Na2SO4), filtered, concentrated, and chromatographed on silica gel to afford the title compound.
c) Ethyl-3-[3 ,4-dihyro-8-iodo-2-thione-5-oxo- 1 H- 1 ,4-benzodiazepine~-4-propanoate To a solution of ethyl-3-[3,4-dihyro-8-iodo-2,5-dioxo- 1 H- 1,4-benzodiazepine]-4-propanoate ( 1.0 g, 2.49 mmol) in THF ( 10 mL) at RT and underan atmosphere of nitrogen is added Lawesson's reagent ( 1.0 g), and the reaction is heated at 50~C for 2 h. The reaction mixture is allowed to cool to RT and is concentrated in vacuo. ~ilica gel chromatography (gradient 40-60%
EtOAc/hexanes) gives the title compound.
d) Ethyl-3-[4H-imi-1~7o~1,2-a][1,4]benzodiazepine-5(6H)-1-methyl-6-oxo-9-iodo]-4-propanoate To a vigorously stirred biphasic solution of ethyl-3-[3,4-dihyro-8-iodo-2-thione-S-oxo-lH-1,4-benzodiazepine]-4-propanoate (0.95 g, 2.27 mmol), methyl iodide (0.2 g) and a catalytic amount of tetrabutylammonium hydrogen sulfate in CH2CI2 (10 rnL) and water (10 mL) is added 2 N NaOH (1.2 mL) at RT. After 2 h, the layers are separated and the aqueous layer is washed with CH2Cl2 (2 x 25 rnL).
The combined organic extracts are dried (Na2SO4), filtered and concentrated in vacuo. The resulting residue is dissolved in toluene (10 rnL) and treated with W O 97/24119 PCTr~S96/20748 pl~pa.~yl amine (0.64 mL, 4-fold excess) and pyridine hydrogen chloride (0.23 g, 1 mol eq). The reaction is heated to reflux for 6 h, then was allowed to cool to RT. Concentration and silica gel chromatography (EtOAc) gives the title compound.
s e) Ethyl-3-[4H-imi~l~7~[1,2-a][1,4]benzodiazepine-5(6H)-l-methyl-6-oxo-9-[[[(benzimidazol-2-yl)methyl]methylamino]carbonyl] }-4-propanoate A rnixture of ethyl-3-[4H-imi~1~7O[ l ,2-a] ~ 1,4]benzodiazepine-5(6H)- 1 -methyl-6-oxo-9-iodo]-4-propanoate(2 mmol), 2-(methylaminomethyl)ben7imi(1z~7c-1e(3 mmol), DIEA (1.8 mL, 10 mmol), and (Ph3P)2PdCl2 (140 mg, 0.2 mmol) in N-methyl-2-pyrrolidinone (20 mL) is heated to 110~C under a CO balloon for 3 h. The mixture is then concentrated and the residue is purified by silica gel flash chromatography to give the title compound.
f) 3-[4H-Imidazo[1,2-a][1,4]benzodiazepine-5(6H)-l-methyl-6-oxo-9-[[[(ben7imi~i~7Ql-2-yl)methyl]methylamino]carbonyl]]4-propanoic acid A solution of ethyl-3-[4H-imi~l~7o[l~2-a][l~4]benzodiazepine-5(6H) methyl-6-oxo-9-[[t(ben7imi~1~7ol-2-yl)methyl]methylamino]carbonyl] }-4-propanoate (54 mmol), LiOH H20 (0.79 mmol), THF (5 mL), and water (2 mL) is stirred at RT overnight. The mixture is concentrated, the residue is dissolved in water, and the resulting solution is neutralized with 3 N HCl. The precipitate is collected and dried in vacuo to afford the title compound.
E~xample 63 P~ lion of 4-r4-r2-(lH-ben7~imi~l~7~ol-2-yl)ethyll-l-piperazin piperi~1inP~çetiç acid a) Ethyl 4-[4-[(tert-butoxycarbonyl)]-1-pipe~ lyl]-1-piperi~1inP~et~t~
The title compound is prepared from tert-butyl l-piperazinecarboxylate (Aldrich) and ethyl 4-oxo-1-piperitlinP~ret:ltP (Porter et al. EPA 0 542 363 A2) by NaCNBH3 reductive amination according to the method of Porter et al., EPA 0 542 363 A2.
b) Ethyl 4-(1-~ipe~dzinyl)- 1-piperi(1in~P;Icet~tP
W O 97/24119 PCTrUS96/20748 A solution of ethyl 4-[4-t(tert-butoxycarbonyl)]-1-pi~e~ yl]-l-piperitlin~a~et~t~ and 4 M HCl/dioxane in CH2Cl2 is stirred at RT for 18 h. The reaction mixture is concentrated to give the title compound as the hydrochloride salt.
5 c) 2-[2-Chloroethyl)]be.n7.imirl~7.ole A solution of 2-ben7.imici:~7.Qleethanol and thionyl chloride in CH2Cl~ is heated at reflux for 2 h. The mixture is evaporated to give the title compound.
d) Ethyl of 4-[4-[2-(lH-bPn7imi<1~7.Ql-2-yl)ethyl]-l-pipeld~ yl]-l-10 piperitlinf.~e~slt~
A solution of ethyl 4-(1-pi~ela~ yl)-1-pipericlinP~ret~te, 2-[2-chloroethyl)]ben7imi/i~7.Qle, and Dl~EA in DMF is stirred at RT for 18 h. The mixture is concentrated and purified by chromatography to give the title compound.
e) 4-[4-[2-(lH-Ben7imi(1~7- 1-2-yl)ethyl]-1-~ .dzinyl]-1-piperi~1inP.~reti~ acidA solution of ethyl of 4-[4-[2-(lH-ken7.imiti~7.--1-2-yl)ethyl]-l-pipera_inyl]-l-piperi~linea~et~te. and 1.0 N NaOH in MeOH is stirred at RT. After 18 h, the mixture is neutralized with AcOH, cle.s~ltP.cl through an XAD-2 column, and lyophiii7P.-l to give the title compound.
F,x~mrle. 64 Prep~ration of 1 -hydroxy-4-r4-r3-( 1 H-be~7imi~1~7.ol-2-yl)propyll- 1 -piperazinyll-eyclohex:~neacetic acid a) tert-Butyl l-hydroxy4-[4-[2-(lH-bçn7.imi-1~7.ol-2-yl)propyl~ ;ldzinyl]-cyclohexaneacetate A solution of tert-butyl l-hydroxy-4-(1-pipel~inyl)-cyclohexs~nP~-~et~tP
(EPA O 537 980 Al~, 2-(3-bromopropyl)ben7imi(1~7.ole (J. Org. Chem. 1962, 27, 3U 2165), and DIEA in DMF is stirred at RT for 18 h. The l~ Lul~iS concentrated and purified by chromatography to give the title compound.
b) 1 -Hydroxy-4-[4-[2-~ lH-ben7.imi~l~701-2-yl)propyl]- l-pipera_inyl]-cyclohex:~n~ etic acid A solution of tert-butyl l-hydroxy~-[4-[2-(lH-ben7imi~1~7.ol-2-yl~propyl]-l-yip~,.dzinyl]-cyclohex~nP.~cet~te and 4 M HCl/dioxane in CH~Cl2 is stirred at RT.
After 18 h, the mixture is evaporated to give the title compound.
CA 0224l633 l998-06-26 W O97/24119 PCTrUS96/20748 Ex~mple 6S
Preparation of N-r3-~l-rr2-(2-Ben7imi(1~7olvl)ethyllcarbonyllpiperidinyllcarbonyll-5 ,13-alanine Following the procedures of Beavers et. al., WO 95/25091, Example 1, except ~ubstitl-tin~ (2-ben7imi~701yl)propionic acid for N~'-Boc-D-lys(Cbz)-OH, gives the title compound.
Example 66 Preparation of 2-r~Ben7imi(1~7ol-2-yl)methyll- 5-r2-~carboxy-ethyl)amino~call,ollyll -2.3-dihydro-3-oxo- 1 H-isoindole Following the procedures of rl~Jaldlion 1-12 in Hartman, et al., EP 0 540 334 Al, for the prepa-dlion of l-H-isoindole-5-carboxamide, 2,3-dihydro-N-(2-carboxy-ethyl)-2-[2-(piperidinyl)ethyl]-3-oxo, except substituting 2-(aminomethyl)be.l~ lidazole (Aldrich) for Boc-4-piperidine-2-ethylamine, the title compound is pl~pal~ed.
Bxample 67 Ple~ lion of r3(R)-r2-(be~ 7ol-2-yl)ethyll-2-oxopiperidinyllacetyl-3(R)-methyl -~-alanine a) Methyl 4-(ben7imifl~7Ol-2-yl) butanoate Following the procedure of Example 36(a), except sub~LiLuLillg 1,2-diaminobenzene for the 2,3-diaminopyridine, the title compound is prepared.
b) 4-(Ben7imi~ 7OI-2-yl) butanoic acid Following the procedure of Example 36(b), methyl 4-(benzimidazol-2-yl) butanoate is saponified to afford the title compoud.
c) [3(R)-r2-~Ben7.imi~ ol-2-yl)ethyl]-2-oxopiperidinyl]acetyl-3(R)-methyl-,13-- 35 alanine .
W O 97/24119 PCT~US96/20748 Following the procedure of Duggan et al (J. Med. Chem. 1995, 38, 3332), except using 4-(ben7imid~7Ol-2-yl) butanoic acid instead of (N-Boc-piperidin-4-yl3butanoic acid, the title compound is prepared.
E~ rr~le68 .
Preparation of 4-rrrr2-(benzimidazolyl)methyllcall,unyllmethylaminol-acetyllphenoxyacetic acida) 4-[2-(Boc-methylamino)acetyl]phenol A solution of di-tert-butyl dicarbonate (5.96 g, 27.3 mmol) in 1,4-dioxane (25 mL) was added dropwise at 0~C to a mixture of 4-t2-(methylamino)acetyl]phenol hydrochloride (5.0 g, 24.8 mmol), 1,4-dioxane (30 mL), H2O (25 rnL) and 1.0 N NaOH (25 mL, 25 mmol). After 24 h, the reaction was warmed to RT and stirred for 1.5 h. More 1.0 N NaOH (25 mL, 25 mmol) was added, and the reaction was stirred for an additional 0.5 h at RT, and concentrated.
The residue was diluted with EtOAc (80 mL), and the ll~ix.Lule was acidified to pH 2 using 1.0 M NaHSO4. The res-llsin~ mixture was extracted with EtOAc, and the combined organic layers were washed with H20 and dried (Na2S04). Filtration and concentration gave the title compound (6.49 g, 99%): lH NMR (250 MHz, CDCl3) ~ 6.70-8.05 (m, 4 H), 4.53 (s, 2H), 2.98 (s, 3H), 1.50 (s, 9H).
b) Benzyl 4-[2-(Boc-methylamino)acetyl]phenoxyacetate A mixture of the compound of Example 68(a) (5.04 g, 19.0 mmol) and K2CO3 (2.63 g, 19.0 mmol) in acetone (100 mL) was stirred at reflux under argon for lh. The mixture was cooled to RT and benzyl bromo~etS-~ (5.23 g, 22.8 mmol) was added. The reaction was heated at reflux for 18 h, then was cooled and filtered.
The filter cake was washed with acetone, and the filtrate was concentrated. The residue was dissolved in CH2Cl2 (300 mL) and washed sequentially with H2O (50 mL) and brine (50 mL). Drying ~Na2SO4), concentration, and flash chromatography ( silica gel, 1 :3 EtOAc/hexanes) yielded the title compound (7.28 g, 93%): 1H NMR (250 MHz, CDCl3) o 6.85-7.95 (m, 9 H), 5.23 (s, 2H), 4.71 (s, 2H), - 4.55 (d, 2H), 2.95 (d, 3H), 1.45 (d, 9H).
c) Benzyl 4-r2-(methylamino)acetyl]phenoxyacetate hydrochloride - A mixture of the compound of Example 68(b) (7.26 g, 17.57 mmol) and 4 M
HCl in 1,4-dioxane (150 mL) was stirred for 1 h at RT. Concentration and trituration with Et2O afforded the title compound as a white powder (5.93 g, 97%):
CA 0224l633 l998-06-26 W O 97/24119 PCTrUS96/20748 1H NMR (250 MHz, CD30D) ~ 7.05-8.00 (m, 9 H), 5.23 (s, 2H), 4.88 (s, 2H), 4.65 (s, 2H), 2.80 (s, 3H).
d) Benzyl 4-~[1:[2-S (bçn7imicl~701yl)methyl3carbonyl]methylalI~ino]acetyl~phenoxyacetate A mi~lule of the compound of Example 68(c)(1 mmol), 2-(ben7imicl~7Olyl)acetic acid (1 mmol), EDC (1.5 mmol), and D~ 3 mrnol) in DMF (25 mL) is stirred at RT. The mixture is poured in to 5% NaHCO3 and extracted with EtOAc. The organic phase is washed with H2O, dried (MgSO4) and concentrated. The residue is chromatographed (silica gel) to give the title compound.
e) 4-[[[[2-(Ben7imit1~7Olyl)methyl]carbonyl]methylamino]acetyl]phenoxyacetate The compound of Example 68(d)(1 mmol) and lN NaOH (1.5 mL) in CH30H (20 mL) is stirred and concentrated. The residue is dissolved in H2O, extracted with CH2C12, and the aqueous phase is adjusted to pH 5 with dilute HCl to give the title compound.
Fx~mI?le 69 Pl~;yaldlion of 4-rrrr2-(Benzimidazolyl)methyllcarbonyllmethylaminolacetyll-1.2-l?henylenedioxvdiacetiç acid a) 4-[2-(Boc-methylamino)acetyl~- 1,2-dihydroxybenzene Following the procedure of Example 68(a), except substituting adrenalone hydrochloride (5.0 g, 23.0 mmol) for 4-[2-(methylamino)acetyl]phenol hydrochloride, the title compound (1.2 g, 19%) was prepared following flash chromatography (silica gel, 1: I EtOAc/hexanes): MS (ES) m/e 282.2 [M+H]+.
b) Dimethyl 4-[2-(Boc-methylamino)acetyl]-1,2-phenylenedioxy~ ret~te Following the procedure of Example 68(b), except substituting the compound of Example 69(a) (0.9 g, 3.2 mmol) for the compound of Example 68(a) and methyl bromoacetate (1.23 g, 8.0 mmol) for ~enzyl bromo~cet~e, the title co.llpoul~d (1.11 g, 81%) was prepared: MS (ES) m/e 426.2 [M+H]+.
c) Dimethyl 4-~2-(methylamino)acetyl]- 1,2-phenylenedioxydiacetate hydrochloride 195 CA 0224l633 l998-06-26 W O 97/24119 PCTrUS96~0748 Following the procedure of Example 68(c), except substituting the compound of Example 69(b) (1.1 1 g, 2.6 mmol) for the compound of Example 68(b), the title colllpoulld was l~r~al~;d (1.1 g, qu~..l ;l~tive): MS (ES) m/e 326.0 [M+H]+.
d) Dimethyl 4-[[[[2-(ben7imi-1A7QIyl)methyl]carbonyl]methylarnino]acetyl]-1,2-phenylenedioxyrliAcetAtP
Following the procedure of procedure of Exarnple 68(d), except sub~ ; ugthe ~;ul~ o~-nd of Example 69(c) for the compound of Example 68(c), gives the title 10 compound.
e) 4-[[[[2-(Benzimidazolyl)methyl]carbonyl]methylamino]acetyl]- 1,2-phenylenedioxydiacetic acid Following the procedure of procedure of l~xarnple 68(e), except substituting 15 the compound of Example 69(d) for the compound of Example68(d), gives the title compound.
Fx~mple 70 Preparatisn of N-r3-rrrt2-Ben7imi-1A7olyl)metllyllçarbonyllamirlolbenzoyll-¦3-alar~ine a) Benzyl N-[3-[[t(2-bçn7imi(1:~7olyl)methyl]carbonyl]arl~ino]benzoyl]-,B-AI~ninAte A ~ Y.lul~ of benzyl N-(3-aminobenzoyl)-,B-~IAninAt~ (Alig, et. al., EPA
372486)(1 mmol), (2-b~7imi~l~7Q}yl)acetic acid (1 mmol), EDC (1.5 mmol), and DIEA (3 rnmol) in DMF (25 mL) is stirred at RT. The mixture is poured into 5%
NaHCO3 and extracted with EtOAc. The combined organic phase is washed with H2O, dried (MgSO4) and concentrated. The residue is chromatographed (silica gel)to give the title compound.
b) N-[3-[[[(2-Bçn7imi~1A7olyl)methyl]carbonyl]amino]benzoylJ-~3-alanine A mixture of the compound of Example 70(a)( 1 mmol) and lN NaOH ( 1.5 mL) in CH30H (20 mL) is stirred and coneell~ldled. The residue is dissolved in H2O, extracted with CH2Cl2, and the aqueous phase is adjusted to pH 5 with dilute HCl to give the title compound.
F.~ 7 1 CA 0224l633 l998-06-26 W O 97/24119 PCTrJS96/20748 Pl~al~tion ofrrl-rN-rr(2-Ben7.imi~7.olyl)methyl]carbonyl~tyrosyll-4-piperidinylloxylacetic acid =
S a) tert-Butyl [[l-[N-[[(2-ben7imi~lA7olyl)methyl]c~ln,llyl]tyrosyl]-4 piperidinyl]oxy]acetate A mixture of tert-butyl [( l-tyrosyl-4-piperidinyl)oxy]acetate (Alig, et. al., EPA 372486)(1 mmol), (2-ben7imi~1A7olyl)acetic acid (1 mmol), EDC (1.5 mmol), and DIEA (3 mmol) in DMP (25 mL) is stirred at RT. The mixture is poured into 10 5% NaHCO3 and extracted with EtOAc. The combined organic phase is washed with H2O, dried (MgSO4) and concentrated. The residue is chromatographed (silicagel) to give the title compound.
b) [[l-[N-[[(2-Benzimida_olyl)methyl]call~ollyl]tyrosyl]~-piperidinyl]oxy]acetic15 acid A mixture of the compound of Example 71(a)(1 mmol) and CF3CO2H in CH2CI2 is stirred and concentrated to give the title compound.
F.x~ ?le 72 P-~pal~lion of fS)-4-rrr(2-Benzimidazolyl)methyllcarbonyllglycyll-3-m~thoxycarbonylmethyl-2-oxopipe-d~ine-l-acetic acid Following the procedure of Sugihara, et. al., EP 0529858, Example 59, except substit~-ting (2-ben7imitlA701yl)acetic -acid for 4-amidinobenzoic acid 25 hydrochloride, gives the title compound.
PxArnple 73 Preparation of (3S.5S)-5-rr4-[(2-Ben71midA7olyl)methyllphenylloxymethyl~-3-30 carboxymethyl-2-pyrrolidinone a) 4-[(2-Ben7imi-1A7l-1yl)methyl]phenol Pollowing the general procedure of Wahlgren and Addison, J. Heterocycl.
- Chem., 1989, 26, 541-543, except substitl-ting 4-(hydroxy)phenylacetic acid for 2-35 (hydroxy)phenylacetic acid, gives the title compound.
..
-CA 02241633 1998-06-W O97/24119 PCT~US96/20748 b) (3S,5S)-5-t[4-[~2-Benzimidazolyl)methyl]phenyl3O~y~ hyl]-3-t(tert-butoxycarbonyl)methyl] -2-pyrrolidinone Following the procedure of Hirnmelsbach, et.al., Australian Patent Application AU-A-86926/91, Example 51, substituting the compound of Example 5 73(a) for 4'-cyano-3'-fluoro-4-hydroxy)biphenyl, gives the title compound.
c) (3S,SS)-5-[~4-[(2-Ben7imi-1~7olyl)methyl~phenyl]oxymethyl]-3-carboxymethyl-2-pyrrolidinone The compound of Example 73(b) is treated with CF3CO2H in CH2CI2 to give the title compound.
F~ le 74 Prçparatioll of 1-r~2-Ben7imi~ m~tllyll-3-r4-(2-carboxyethyl)phenyl~-1 5 m~,thoxy-3-pyrrolin-2-one Following the procedures of Linz, et. al., EP 0567968, except sub~ (2-ben7imi-1~701yl)methsln:~min/ for 4-cyano~nilin~, gives the title colllpo~l,ld.
Fx~ ple 75 Plti~,a,~tion of 2-r6-(ben7imi~1~7ol-2-yl) metllylaminocarbonyl)-12.3 tetrahydroisoquinolinyllacetic acid a) 6-Methoxy- 1,2,3 ,4-tetrahydroisoquinoline 6-Methoxy-1,2,3,~tetrahydroisoquinoline is prepared according to the method of D. J. Sall and G. L. Grunewald (J. Med. C~hem. 1987, 30, 2208-2216).
b) Ethyl 2-[6-Methoxy-1,2,3,~tetrahydroisoquinolinyl]acetate A solution of the compound of Example 75(a) ( 1.1 mmol), ethyl chloroacetate (1.17 mmol), and pot~ssillm carbonate (1.17 mrnol) in acetonitrile (10 mL) is stirred for 18 hr. The mixture is then partitioned in a mixture of EtOAc and H20. The organic phase is rotary evaporated to an oil, which is purified by silica gel chromatography to afford the title co~ oulld.
W O 97/2411g PCTrUS96/20748 c) Ethyl 2-[6-Hydroxy-1,2,3,4-tetrahydroisoquinolinyl]acetate A solution of the compound of Example 75(b3 (0.249 g, 1.0 mmol) and boron tribromide ( lM in CH2Cl2, 1.0 mL, 1.0mmol, mL) is stirred at -70 ~C for 2 hr and then stirred at RT for 12 hr. The solution is rotary evaporated to an oil. The - S residue is taken up in EtOAc. EtOAc is washed with water (lX), 5% NaHCO3(2X), water (lX). EtOAc is dried over Mg2SO4, filtered and rotary evaporated to afford the title compound.
d) Ethyl 2-[6-trifluoromethylsulfonyloxy-1,2,3,4-tetrahydroisoquinolinyl]acetateA solution of the compound of Example 75(c) (0.235 g, 1.0 mmol), trifluorosulfonic acid anyhdride (0.23 mL, l.lmmol,) and Et3N (0.32 mL, 1.5 mmol) in CH2Cl2 (S ml)is stirred for 8 hr The solution is rotary evaporated to an oil. The residue is taken up in EtOAc. EtOAc is washed with 5% NaHCO3 (2X), water (lX).
EtOAc is dried over Na2SO4, filtered and rotary evaporated to afford the title lS compound.
e) Ethyl 2-[6-carboxy-1,2,3,4-tetrahydroisoquinolinyl~acetate A solution of the compound of Example 75(d) (0.367 g, 1.0mmol), pall~ m(II)bis-acetate (0.022 g, 0.1mmol,), triphenylphosphine (0.262 g, 1.0mmol), diisopropylamine (0.34 mL, 2.5 mmol), NMP (S mL), in aqueous ~mmonillm carbonate (10%) is stirred for 8 hr under an atmosphere of carbon monoxide. The solution is rotary evaporated to an oil. The residue is purified by silica gel chromatography to afford the title compound.
f) Ethyl -[6-(ben7imicl~7Ol-2-yl)methylaminocarbonyl)-1,2,3,4-tetrahydroisoquinolinyl]acetic acid A solution of the compound of Example 75(e) (0.263 g, 1.0 mmol), the compound of (2-ben~ 7Olyl)acetic acid (0.34g, 1.0 mmol), EDC (0.19lg, 1.0 - mmol), HOBt (0.151 g, 1.0 mmol) and triethylamine (0.235 mL, 2.0 mmol) in DMF(7 mL) is stirred for 8 hr. The solution is rotary evaporated to an oil. The }esidue is purified by silica gel column to afford the title compound.
CA 0224l633 l998-06-26 W O97/24119 PCTrUS96/20748 g) 2-[6-(ben7imi~1~7ol-2-yl)methylarninocarbonyl)-1,2,3,4-tetrahydroisoquinolinyl]acetic acid A solution of the compound of Example 75(f) ( 0.42 g, 1.0 mmol) in aqueous 1 N sodium hydroxide (1.5 mL, 1.5 mmol) and ethanol ( 5 mL) is stirred for 8 hr.The solution is rotary evaL,ol~led to an oil. The residue is purified by silica gel column to afford the title compound.
Example 76 Pre~?~ration of 2-r6-fbell~i...id~7ol-2-yl)methylaminoca~l,ollyl)-l-oxo-1.2.3~4-tetrahydroisoquinolir~ acetic acid a) 6-Methoxy- 1 -oxo- 1,2,3,4-tetrahydroisoquinoline 6-Methoxy-l-oxo-1,2,3,4-tetrahydroisoquinoline is p~ d according to the method of D. J. Sall and G. L. Grunewald, J. Med. Chem. (1987), 30, 2208-2216.
b) Ethyl 2-[6-Methoxy-l-oxo-1,2,3,4-tetrahydroisoquinolinyl]acetate A mixture of the compound of Example 76(a) (0.39 mmol) and NaH (0.17 g, 0.43 mmol, 60% oil dispersion) in THF (5 rnL) is heated to reflux for 1 hr and then allowed to cool to room ~e~ el~ture. Ethyl chloroacetate (0.43 mmol) is added tothe mixture, and the mixture is allowed to stir for 1 hr. The mixture is quenched with water (10 rnL) and washed with EtOAc (2X 15 rnL). The organic layers are combined, washed with water (10 rnL) and rotary evaporated to an oil. The residue is purified by silica gel chromatography to afford the title compound.
c) Ethyl 2-[6-Hydroxy-l-oxo-1,2,3,4-tetrahydroisoquinolinyl~acetate A solution of the compound of Example 76(b) (0.263 g, 1.0 mmol) and boron tribromide (lM solution in CH~Cll, 1.1 mL) is stirred at -70 ~C for 2 hr and then at RT for 4 hr. The solution is rotary evaporated to an oil. The residueis taken up in EtOAc. EtOAc is washed with water (lX), 5% NaHCO3 (2X), water (lX).
W O97/24119 PCTrUS96/20748 EtOAc is dried over MgSO4, filtered and rotary evaporated to afford the title compound.
~. d) Ethyl 2-[6-trifluoromethylsulfonyloxy-1-oxo-1,2,3,4-- 5 tetrahydroisoquinolinyl]acetate A solution of the compound of F.~mple 76(c) (3.4 mmol) and trifluorosulfonic acid anyhdride (3.4 mmol, mL) in pyridine (5 mL) is chilled at 0~
and allowed to warm to room temperature for 1 hr. The mixture is quenched with water (5 mL~ and washed with EtOAc (2X 7 mL). The organic layers are combined, washed with water (7 rnL) and rotary evaporated to an oil. The residue is purified by silica gel chromatography to afford the title compound.
e) Ethyl 2-[6-carboxy-1-oxo-1,2,3,4-tetrahydroisoquinolinyl~acetate A solution of the compound of Example 76(d) (0.23 g, 1.0 mmol), palladium(II)bis-acetate (0.026 g, 0.1 mmol), triphenylphosphine (0.262 g, 1.0 mmol), diisopropyla~nine (0.23 mL, 2.0 mmol), NMP (7 mL), in aqueous ammonium carbonate "(10 %) is stirred for 8 hr under an atmosphere of carbon monoxide. The solution is rotary evaporated to an oil. The residue is purified by silica gel chromatography to afford the title compound.
f) Ethyl -[6-(bc~ 7Ol-2-yl)methylaminocarbonyl)- 1,2,3,4-tetrahydroisoquinolinyl]acetic acid A solution of the compound of Example 76(e) (0.34 g, 1.0 mmol), the compound of (2-ben~ idazolyl)acetic acid (0.43 g, 1.0 mmol), EDC (0.191 g, 1.0 mmol), HOBt (0.15 g, 1.0 mmol) and triethylamine (0.234 mL, 2.3 mmol) in DMF(8 mL) is stirred for 8 hr. The solution is concentratred. The residue is purified by silica gel chromatography to afford the title compound.
~lt~ tively, a solution of the compound of Example 76(d) (0.23 g, 1.0 mmol), p~ m(II)bis-acetate (0.026 g, 0.1 mmol), triphenylphosphine (0.262 g, - 1.0 mmol), diisopropylarnine (0.25 mL, 2.1 mmol), NMP (7 mL), and the compound W O 97/24119 PCTrUS96/20748 of Interm~ t~ A (0.31 g, 1.0 mmol) in aqueous ammonium carbonate ~10%) is stirred for 8 hr under an atmosphere of carbon monoxide. The solution is rotary evaporated to an oil. The residue is purified by silica gel chromatography to afford the title compound.
g) 2-[6-((ben7imi~l~7ol-2-yl)methylaminocarbonyl)-1,2,3,4-tetrahydroisoquinolinyl]acetic acid A The solution of the compound of Example 76(f) (0.25 g, 1.0 mmol) in aqueous 1 N sodium hydroxide (1.5 mL, 1.5 mmol) and ethanol (8 mL) is stirred for 10 8 hr. Solution is rotary evaporated to an oil. The residue is purified by silica gel to afford the title compound.
Rx~mple 77 Preparation of 2-r6-((be~ 7~1-2-yl)methylcarbonyl:~mino)tetralinlacetic acid a ) tert-butyl-5-amino-tetraline-2-acetate Tert-butyl-5-amino-tetraline-2-acetate is prepared according to the methods described in M. J. Fisher, et al. (Scheme 12 and Example 28, parts A-D, EO
0635492, Jan. 25, 1995).
b) 2-[6-((benzimidazol-2-yl)methylcarbonylamino)tetralin]acetate A solution of the compound of Example 77 (a) (0.261 g, 1.0 mmol), 2-(aminomethyl)ben7imid~7O1e(0.256g, l.Ommol),EDC(0.19lg, l.Ommol) 1-hydroxybenzotriazole hydrate (0.152 g, 1.0 mmol), and triethylamine (0.234 mL, 2.1 mmol) in DMF (5 mL) is allowed to stir for ~ hr. The solution is rotary evaporated to an oil. The residue is purified by silica gel chromatography to afford the title compound.
A solution of the crude ester amide (0.32 g, 1.0 mmol) and trifluoroacetic acid (5 mL) in methylene chloride (~ mL) is allowed to stir for 1 hr. The solution is CA 0224l633 l998-06-26 W O97/24119 PCT~US96/20748 rotary evaporated to an oil. The residue is treated with Et2O. Filtration and drying in vacuo afforded the title compound.
;, F.x~mple 78 ~l~;palalion of 2-r6-((benzimidazol-2-yl)methylaminocarbonyl)tetr~linJacetic acid a) Ethyl-5-hydroxy-tetraline-2-acetate The compound Ethyl-5-hydroxy-tetraline-2-acetic acid is prepared according to the method of M. J. Fisher et al. (EP 0635492, Scheme 6 and Example 20, parts A-D, p. 71).
b) Ethyl-5-trifluoromethylsulfonyloxy-tetraline-2-acetate A solution of the compound of Example 78(b) (0.321g, 1.0 mmol) in CH2Cl~
(10 mL) is cooled to O~ C.Trifluoromethylsulfonic acid anhydride ( 0.125 mL, 1.1mmol) is added. The solution is stirred for 2 hr. The solution is rotary evaporated to an oil. The residue is taken up in EtOAc. EtOAc is washed with water (lX), 5%
NaHCO3, water (lX). EtOAc is dried over MgSO4, filtered. Filtrate is rotary evaporated to afford the title compound.
c) Ethyl 6-carboxy-tetraline-2-acetate.
A solution of the compound of Example 78(c) (0.26 g, 1.0 mmol), palladium(II)bis-acetate (0.023g, 0.1 mmol), triphenylphosphine ~0.262g, 1.0 mmol), diisopropylamine (0.245 mL, 2.1 mmol), NMP (10 mL ), in aqueous 25 ammonium carbonate (10%) is stirred for 8 hr under an ~tmosphere of carbon monoxide. The solution is rotary evaporated to an oil. The residue is purified by silica gel chromatography to afford the title compound.
- d) Ethyl -[6-((ben7imi~l~7ol-2-yl)methylaminocarbonyl)-tetraline-l-acetate A solution of the compound of Example 78(c) (0.34 g, 1.0 mmol), the compound of (2-ben7imicl~olyl)acetic acid (0.32g, 1.0 mmol), EDC (0.191 g, 1.0 W O 97/24119 PCTrUS96120748 mmol), HOBt ~0.152 g, 1.0 mmol) and triethylamine (0.23 mL, 2.1 mrnol3 in DMF(6 mL) is stirred for 8 hr. The solution is rotary evaporated to an oil. The residue is purified by silica gel chromatography to afford the title compound.
~It.orll~tively, a solution of the compound of Example 78(b) ~0.34 g, 1.0 mmol), palladium(II)bis-acetate (0.023g, 0.1 mmol), triphenylphosphine (0.262g, 1.0 mmol), diisopropylarnine (0.23 mL, 2.1 mmol), NMP (10 mL), and the compound of IntermPrli~te A (0.32 g, 1.0 mmol), in aqueous ammonium carbonate (10%) is stirred for 8 hr under an zitm~spht~re of carbon monoxide. The solution is rotary evaporated to an oil. The residue is purified by silica gel chromatography to afford the title compound.
e) 2-~6-((be.~i."~ 7Ol-2-yl)methylaminocarbonyl)-tetraline-2-acetic acid.
A solution of the compound of Example 78(d) (0.31 g, 1.0 mmol) in aqueous 1 N sodium hydroxide (1.5 mL, 1.5 mmol) and ethanol ( 5 mL) is stirred for 8 hr.The solution is rotary evaporated to an oil. The residue is purified by silica gel chromatography to afford the title compound.
Fx~mple 79 Preparation of 2-r5-((be~7,i l ll;dzl~ol-2-yl)methylcarbonylamino)-benzofuranl-.
proplomc acld a) Ethyl 1-carboxymethyloxy~-nitrosalicylaldehyde A solution of 1-hydroxy~-nitrosalicylaldehyde (Aldrich) ( 0.167 g, 1.0 mmol), ethyl bromoacetate (0.166 g, 1.0rnmol) potSlc~inm carbonate ( 0.276 g, 2.0mmol) and sodium iodide (0.015 g, 0.1 mmol) in THF ( 10mL) is heated to 80 ~Cfor 24 hr. The solution is rotary evaporated to an oil and the residue is purified by silica gel chromatography to afford the title compound.
b) Ethyl 2-carboxy-5-nitrobenzofuran W O 97/24119 PCTrUS96/20748 A solution of the compound of Example 79(a) (0.229 g, 1.0 mmol), DBU (0.152 g, 1.0 mmol) in ethyl alcohol ( 10 mL) is allowed to stir at RT for 18 hr.
The solution is rotary evaporated to an oil and the residue is treated with EtOH (10 mL). The solution is bubbled with HCl.gas for 2 minllt~s and refluxed for 5 hr. The - 5 solution is rotary evaporated to an oil. EtOAc is added and washed with water (2X), 5% citric acid (2X), water (1~), 5% NaHCO3.(2X) and water (lX). EtOAc is rotary evaporated to afford the title compound.
c) Ethyl 2-(2-carboxy)ethylene-5-nitrobenzofuran A cold solution (-78 ~C) of the compound of Example 79(b) ( 0.235 g, 1.0 mmol)in THF (5 mL) is treated with DiBAL ( 1.0 M in THF, 1.0 mL, 1.0 mmol).
The solution is stirred at -78 ~C for 30 min~ltes and RT for 3 hr. The solution is treated with CH3COOH (3 mL) followed by water (2 rnL). The solution is rotary evaporated to an oil and treated with toluene to azeotrope off the acetic acid. Drying in vacuo afforded the crude aldehyde.
A solution of the phosphonate ester (0.224 g, 1.0 mmol),in THF (5 mL) is treated with sodium hydride (60% suspension in mineral oil, 0.04 g, 1.0 rnmol)at 0~
C for 1 hr. To the solution is added the aldehyde (0.235 g, 1.0 mmol). The solution is stirred at RT for 18 hr. The solution is rotary evaporated to an oil and the residue is purified by silica gel chromatography to afford the title compound.
d) tert-butyl 2-[5-amino-benzofuranyl]propionate A solution of the compound of Example 79(c) ( 0.261 g, 1.0 rnrnol) in ethanol ( 5 mL) containing 10% palladium-on-carbon(0.026 g, 10% wt) is hydrogenated at 45 psi for 1 hr. The solution is filtered through Celite and the- filtrate is rotary evaporated to an oil. Silica gel cl~ t~ graphy affords the title compound.
e) 2-~5-((bçn7imicl~7Ql-2-yl)methylcarbonylamino)-benzofuran]-propionic acid W O 97/24119 ~CT~US96/20748 A solution of the compound of Example 79(d) ( 0.263 g, 1.0 mmol), EDC ( 0.191 g, 1.0 mmol), 1-hydroxybenzotriazole ( 0.150 g, 1.0 mmol), the compound of 2-(aminomethyl)benzimidazole ( 0.234 g, 1.0 mmol) and triethylamine (0.288 mL, 2.0 mmol) in DMF (5.0 mL) is allowed to stir for 18 hr. The solution is rotary evaporated to an oil and the residue is purified by silica gel chromatography to afford the title compound.
A solution of the crude ester ( 0.263 g, 1.0 mmol) in MeOH (3.0 mL) is treated with 1 N NaOH ( l .S mL, l .S mmol) and water (2 ml). The solution is stirred at RT for 18 hr. The solution is rotary evaporated to an oil and purified by reversed phase chromatography to afford the title compound.
F.x~?le 80 Preparation of 2-rS-((benzirnidazol-2-yl)methylcarbonylamino)-2.3-dihydro-ben7Ofilranl-propionic acid a) Ethyl-2-[S-amino-2,3-dihydro-benzofuranyl3propionate In the chromatographic purification of the compound of Example 79(d) the -20 title compound is also obtained.
b) 2-[5-(6-arninopyridinyl-2-methylcarbonylamino)-benzofuran]-propionic acid A solution of the compound of Example 80(a) (0.263 g, 1.0 mmol), EDC
(0.19lg, 1.0 rnmol), 1-hydroxybenzotriazole (0.lSg, 1.0 m~nol), the compound of 2-(aminomethyl)ben7imi.1~701e (0.32 g, 1.0 mmol) and triethylamine (0.288 mL, 2.0 mmol) in DMF (5.0 mL) is allowed to stir for 18 hr. The solution is rotary evaporated to an oil and the residue is purified by silica gel chromatography toafford the title compound.
A solution of the crude ester ( 0.290g, 1.0 rnmol) in MeOH (5.0 mL) is treated with lN NaOH (1.2 mL, 1.2 mmol) for 18 hr. The solution is rotary .
W O 97/24119 PCT~US9~/20748 evaporated to an oil and purified by reversed-phase chromatography to afford the title compound.
-Fxample 81 S
Preparation of 2-r5-(6-aminopyridinyl-2-nlethylaminocarbonyl)-benzofuranl-propionic acid a) 2-Ethyloxycarbonyl-5-(tert-butyl-dimethylsilyloxy)-benzofuran A solution of 2-Ethyloxycarbonyl-5-(hydroxy)-benzofuran is prepared via the procedure described in M. L. Denny, et al. (EP 0655439, 31,5,95) (0.206 g, 1.0 mmol), tert-butyl-dimethylsilylchloride (0.23 mL, 1.0 mmol) and imidazole ( 0.34 g, 1.0 mmol) in THF is allowed to stir for 4 hr. The solution is rotary evaporated to an oil. EtOAc is added and washed with water. EtOAc is rotary evaporated to afford the title compound.
b) 2-Hydrocarbonyl -5-(tert-butyl-dimethylsilyloxy)-benzofuran A cold solution (-78 ~ C) of the compound of Example 81 (a) ( 0.35 g, 1.0 mmol)in THF (5 mL) is treated with DiBAL ( 1.0 M in THF, 1.0 mL, 1.0 mmol).
The solution is stirred at -78 ~ C for 30 minutes and RT for 3 hr. The solution is treated with CH~COOH (3 mL) followed by water (2 mL). The solution is rotary evaporated to an oil and treated with toluene to azeotrope off the acetic acid. Drying in vacuo afforded the aldehyde.
c) Ethyl 2-[5-(tert-butyl-dimethylsilyloxy)-benzofuran]-acrylate A solution of the phosphonate ester (0.224 g, 1.0 mmol),in THF (5 rnL) is treated with sodium hydride (60% s-lcpen~ion in mineral oil, 0.04 g, 1.0 mmol)at 0"
C for 1 hr. To the solution is added the above aldehyde(0.235 g, 1.0 mmol). The solution is stirred at RT for 18 hr. The solution is rotary evaporated to an oil and the residue is purified by silica gel chromatography to afford the title compound.
W O 97/24119 PCT~US96J20748 d) 2-Ethyl 2-~-5-(hydroxy)-benzofuran]-propionate A mixture of the compound of Example 81(c) (0.234g, 1.2 mmol) and 10%
p~ m-on-carbon (0.023 g, 10% wt) in EtOH( S mL).is hydrogenated at 50 psi 5 for 1 hr. Filtration through Celite and concentration afforded.the ester (0.169 g, 56%).
A solution of the crude ester (0.34 g, 1.0 mmol) and tetraethylammonium fluoride (0.149 g, 1.0 mmol) in THF (10 mL) is allowed to stir at RT for 18 hr. The 10 solution is rotary evaporated to an oil and purified by silica gel ch~ atography.
e) 2-Ethyl 2-[-5-(trifluoromethylsulfonyloxy)-benzofuran]-propionate A solution of the compound of Example 81(d) (0.366 g, 1.0 mmol) and Et3N(0.23 mL, 1.5 mmol) in CH2Cl2 (10 mL) at 0 "C is treated with trifluoromethylsulfonic acid anhydride (0.21 mL, 1.1 mmol). After 2 hr solution is rotary evaporated to an oil. The residue is taken up in EtOAc. EtOAc is washed successively with water(lX), 5% Na~ICO3 (2X), water (lX). EtOAc is dried over MgSO4, and filtered. Filtrate is rotary evaporated to afford the title compound.
20 f) 2-Ethyl 2-[-5-(carboxy)-benzofuran3-propionate A solution of the compound of Example 81(e) (0.366g, 1.0 mmol), p~ m(II) bis-acetate (0.023g, 0.1 mmol), triphenylphospine (0.262g, 1.0 mmol), diisopropylethylamine (0.23 mL, 2.1 mmol), NMP (7 mL) in aqueous sodium bicarbonate (10%, 6 mL) is allowed to stir. The solution is rotary evaporated to an 25 oil. The residue is purified by silica gel chromatography to afford the title compound.
g) 2-Ethyl 2-~-5-(6-(6-aminopyridinyl-2-methylaminocarbonyl)-benzofuran]-propionate W O 97/24119 PCT~US96/20748 A solution of the compound of Example 81(f) (0.366 g, 1.0 mmol), the compound of (2-ben7imid~701yl)acetic acid (0.23g, 1.0 mmol), EDC (0.191 g, 1.0mmol), HOBt (0.152g, 1.0mmol) and triethylamine (0.235 mL, 2.1mmol) in DMF(8 mL) is stirred for 8 hr. The solution is rotary ev~poldled to an oil. The 5 residue is purified by silica gel chromatography to afford the title compound.
Alt~-rn~tively, a solution of the compound of Example 81(e) (0.366g, l .Ommol), palladium(II)bis-acetate (0.023g, 0.1 mmol), triphenylphosphine (0.262g, 1.0 mmol), diisopropylamine (0.23 mL, 2.1 mmol), NMP (10 mL ), and the compound of Intern~ t(~ A (0.32g, 1.0 mmol) in aqueous ammonium carbonate (10%, 10 mL) is stirred for 8 hr under an atmosphere of carbon monoxide. The solution is rotary evaporated to an oil. The residue is purified by silica gel chromatography to afford the title compound.
h) 2-[-5-(6-(6-aminopyridinyl-2-methylaminocarbonyl)-benzofuran]-propionic acid A solution of the col"pou~d of Example 81(g) (0.366g, 1.0 mmol) in aqueous 1 N sodium hydroxide (1.5 mL, 1.5 mmol) and ethanol ( 8 rnL) is stirred for 8 hr. The solution is rotary eva~o,~l~d to an oil. The residue is purified by silica gel chromatography to afford the title compound.
Ex~n~le 8~
Preparation of 2-rS-((benzi",idazol-2-yl)methylaminocarbonyl)-2.3-dihydro-ben7:r~furan~-propionic acid a) 2-tert-Butyl 2-[-5-(hydroxy)-2,3-dihydro-benzofuran]-propionate The chromatographic purification in Example 81 (d) also provides the title compound.
b) 2-tert-Butyl 2-[-5-(trifluoromethylsulfonyloxy)-2,3-dihydro-benzofuran]-propionate W O 97/24119 PCT~US96/207~.
A cold solution of the compound of Example 82(a) (0.28 g, 1.0 mmol) and Et3N (0.23 mL, 2.1 mmol) in CH2Cl2 (5 mT.)is treated with trifluoromethylsulfonic acid anhydride (0.15 mL, 1.1 mmol) for 2 hr. The solution is rotary evaporated to an oil. The residue is taken up in EtOAc. EtOAc is washed 5 with water (lX), 5% NaHCO3(2X), water (lX). EtOAc is dried over MgSO4, filtered, and filtrate is rotary evaporated to afford the title compound.
c) 2-tert-Butyl 2-[-5-(carboxy)-2,3-dihydro-benzofuran]-propionate A solution of the compound of Example 82(b) (0.24 g, 1.0 mmol), 10 p~ um(II) bis-acetate (0.023g, 0.1 mmol), triphenylphospine (0.262g, 1.0 mmol), diisopropylethylamine (0.23 rnL, 2.1 mmol), NMP (8 mL ) in aqueous sodium bicarbonate (10 mL, 10%) is allowed to stir at RT for 8 hr. The solution is rotary evaporated to an oil. The residue is purified by silica gel chromatography to afford the title compound.
d) 2-tert-Butyl 2-[-5-(6-(6-aminopyridinyl-2-methylaminocarbonyl)-2,3-dihydro-benzofuran] -propionate A solution of the compound of Example 82(c) (0.366g, 1.0 mmol), the compound of (2-be~ 7olyl)acetic acid ( 0.23g, 1.0 mmol), EDC (0.19lg, 1.0 20 mmol), HOBt (0.152g, 1.0 ~nol) and triethylamine (0.23 mL, 2.1 mmol) in DMF(8mL) is stirred for 8 hr. The solution is rotary evaporated to an oil. The residue is purified by silica gel chromatography to afford the title compound.
~It~ tively, a solution of the compound of Example 82(b) (0.366g, 1.0 25 mmol), p~ m(II)bis-acetate (0.023 g, 0.1 mmol), triphenylphosphine (0.2~2 g, 1.0 rnmol), diisopropylamine (0.23 mL, 2.1 mmol), NMP (10 mL), and the compound of Interm~ te A (0.23 g, 1.0 mmol), in aqueous ammonium carbonate (10%, 10 mL) is stirred for 8 hr under an atmosphere of carbon monoxide for 8 hr.
The solution is rotary evaporated to an oil. The residue is purified by silica gel 30 chromatography to afford the title compound.
W O97/24119 PCT~US96/20748 e) 2-[-5-(6-((ben7imit1~7Ql-2-yl)methylaminocarbonyl)-2,3-dihydro-benzofuran]-propionic acid . A solution of the compound of Example 82(d) (0.37 g, 1.0 mmol) in aqueous 1 N sodium hydroxide ( 1.5 mL, 1.5 mmol) and ethanol (8 mL) is stirred for 8 hr.- 5 The solution is rotary eva~oldted to an oil. The residue is purified by silica gel chromatography to afford the title compound.
.
Example 83 lQ ~lcpald~ion of 1 -r( lH-ben7imi(1~7ol-2-yl)methyll-3-{4r(2-ethoxycarbonyl)ethyl)lphenyl }-3-oxo-imidazolidine a) 2-Aminomethyl-l-p-toll-enl-sl-lfonyl-benzimidazole A solution of 2-Methylben7imi~701e (1 mmol), p-toluenesulfonyl chloride (1.05 mmol) and Et3N (1.05 mmol) in water (2 mT ) and THF (1 mL) is allowed to stir for 4 hr. The ~ L~nc is concentrated by rotary evaporation and diluted withwater (10 mL). The solution is washed with EtOAc (2X 15 mL). The organic layers are combined, washed with water (S mL) and rotary evaporated to an oil.
A solution of the aforementioned oil (2-Methyl- 1 -p-toluenesulfonyl-benzimidazole) (1 mmol) and N-bromosuccinimide (1.05 mrnol) in CCl4 (10 mL) is allowed to reflux for 18 hr. Upon cooling a white solid precipitates from solution.
The solid is filtered, triturated with CCl4 and dried in vacuo to a solid.
A solution of Boc2NH (1 mmol) and pot~sil-m hydroxide ~1 mmol~ in ethanol (S mL) is allowed to stir for 1 hr. Anhydrous ethyl ether (15 mL) is added and the mixture is filtered to afford the salt as a white solid. A solution of the aforementioned solid 2-bromomethyl-l-p-toluenesulfony!bl~n7i..~ 701e (1 mmol) and (Boc)2N~ Kt (mmol) in THF (10 mT.)is to stir at 60~ for 18 hr. The mixture is 30 rotary evaporated to an oil.
A solution of the 2-[bis-(tert-butyloxycarbonyl)aminomethyl]- l-p-toluenesulfonylben7im~ 701e (1 mmol) in TFA (1.1 mmol) and CH2Cl2 is allowed to stir for 1 hr. The solution is rotary ev~o-dled to an oil, which is purified by 35 chromatography to afford the title compound.
W O97/24119 PCT~US96/20748 b) Ethyl 2-[4-(2-hydroxyethylamino)phenyl]propionate This compound is prepared following the procedure of F. ~imm~l~bach et al.
~Example V, p. 44, EP 0587134, Sept. 8, 1993), in which glycolaldehyde dimer (Aldrich) (mmol) is added to a solution of methyl 2-(4-aminophenyl)propionate 1 (1 mmol) in aqueous acetonitrile (pH 6-7) (mL), followed by sodiumcyanoborohydride (1.1 mmol), and the mixture is allowed to stir for 1 hr. The lluhLIul~ is rotaryevaporated to an oil, and the residue is dissolved in a mixture of ice water and ethyl acetate. The water layer is neutralized with 4 N sodium hydroxide and washed with ethyl acetate. The organic phase is rotary evaporated to an oil. A solution of the oil in ethyl acetate is purified on a silica gel column to give the title compound.
c) N-[(l-p-toluenesulfonyl-lH-be~ 7~l-2-yl)methyl]-N~-hydroxyethyl-N~
{4[(2-ethoxycarbonyl)ethyl)]phenyl } -urea This compound is Lnc~cd following the procedures of F. Himmelsbach et al. (EP 0587134, Sept. 8, 1993 and EP 0612741, Feb.21, 1994), in which a solution of the compound of Example 83(a) (1 mmol) and phosgene (1.1 mmol) in THF (20 mL) is allowed to stir at -20~ for 20 minutes The compound of Example 83(b) (1.0mmol) is added to the solution and the r~cllltin~ mixture is allowed to stir for 18 hr.
The resulting solution is rotary evaporated. A solution of the residue in ethyl acetate is washed with 5% citric acid, followed by water. The organic phase is rotary evaporated to an oil. . A solution of the oil in ethyl acetate is purified on a silica gel column to give the title compound.
d) N'-[(l-p-toluenesulfonyl-lH-ben7imifl~7ol-2-yl)methyl]-N3-{4t(2-ethoxycarbonyl)ethyl)]phenyl } -2-oxo-imidazolidine This compound is prepared following the procedures of F. Himmelsbach et al. (Example III, EP 0587134, Sept. 8, 1993 and EP 0612741, Feb.21, 1994), in which a solution of the compound of Example 83(c) (1 mmol), mf-th~n~sulfonylchloride (1.2 mmol) and triethylamine (1.2 mrnol) in methylene chloride (5 ml) is allowed to stir at 0~ for 1 hour. The ~ lul~; is partitioned in a mixture of water and methylene chloride. The organic phases are combined and dried over anhydrous sodium sulfate and rotary evaporated.
A solution of the residue and sodium iodide (1.1 mmol) in acetone (5 mL) is heated to reflux for 3 hr and then rotary evaporated to an oil. Potassium-bis(trimethylsilyl)azide (1.2 mmol) is added to a solution of the residue in DMF (5 mL), cooled to 0~. The solution is allowed to warrn to room t~ln~el~lul~ over 30 212 , PCT~US96120748 min. and the rotary evaporated to an oil. The residue is partitioned in a ul~ of water and methylene chloride. The organic phases are combined and dried over anhydrous sodium sulfate and rotary evaporated. A solution of the oil in ethyl acetate is purified on a silica gel column to give the title compound.
e) N~ H-bçn7im~ 7ol-2-yl)methyl]-N3-{4[(2-carboxyl)ethyl)]phenyl}-2 imicl~olidine A solution of the compound of Example 83(d) ( 1 mmol) in THF (S mL) and 1 N sodium hydroxide (1.2 mL, 1.2 mmol) is allowed to stir for 18 hr. The mixture 10 is neutralized with conc. hydrochloric acid and purified on a silica gel column to give the title compound.
F,~ m,ple 84 Parenteral Dosage Unit Composition A preparation which contains 20 mg of the compound of Example 1 as a sterile dry powder is prepared as follows: 20 mg of the compound is dissolved in 15 mL of distilled water. The solution is filtered under sterile conditions into a 25 ml multi-dose ampoule and lyophilized. The powder is reconctit-lte~l by addition of 20 mT of 5% dextrose in water (DSW) for intravenous or intr~mllccul~r injection. The dosage is thereby det~,rmin~l by the injection volume. Subsequent dilution may be made by addition of a metered volume of this dosage unit to another volume of DSW for injection, or a metered dose may be added to another m~c,h~nicm for en~ g the drug, as in a bottle or bag for IV drip infusion or other injection-infusion system.
Exarnple 85 C)ral Dosa~e Unit Composition A capsule for oral ~f1mini,ctration is prepared by mixing and rnilling 50 mg of the compound of Example 1 with 75 mg of lactose and 5 mg of m~gnl~,citlm stearate.
The rçs--ltin~ powder is screened and filled into a hard gelatin capsule.
F,x:~mple 86 Oral r~osa~e Unit Com,~osition A tablet for oral a~1minictration is prepared by mixing and gr~n~ ting 20 mg of sucrose, 150 mg of calcium sulfate dihydrate and 50 mg of the compound of J
CA 0224l633 l998-06-26 Example 1 with a 10% gelatin solution. The wet granules are screened, dried, mixed with 10 mg starch, 5 mg talc and 3 mg stearic acid; and co~ r~ssed into a tablet.
The above description fully discloses how to make and use the present - invention. However, the present invention is not limited to the particular embodiments described hereinabove, but includes all modifications thereof withinthe scope of the following claims. The various references to journals, patents and other publications which are cited herein comprises the state of the art and areincorporated herein by reference as though fully set forth.
r
Claims (52)
1. A compound according to formula (I) or (II) or (III) or (IV) or (V):
or or or or wherein:
W is - (CHRg)b-V'- or phenyl;
A is a fibrinogen receptor antagonist template;
V' is CONR21 or NR21CO;
G is NRe, S or O;
Rg is H, C1-6alkyl, Het-C0-6alkyl, C3-7cycloalkyl-C0-6alkyl or Ar-C0-6alkyl;
R21 is Het-(CH2)0-6-U'-(CH2)1-6-, C3-7cycloalkyl-(CH2)0-6-U'-(CH2)1-6-, or Ar-(CH2)0-6-U'-(CH2)1-6-;
U' is CONRf or NRfCO;
Rf is H, C1-6alkyl or Ar-C1-6alkyl;
Re is H, C1-6alkyl, Ar-C1-6alkyl, Het-C1-6alkyl, C3-7cycloalkyl-C1-6alkyl, (CH2)qOH or (CH2)kCO2Rg;
k is 0, 1 or 2;
q is 1 or 2;
b is 0, 1 or 2;
Rb and Rc are independently selected from H, C1-6alkyl, Ar-C0-6alkyl, Het- C0-6alkyl, or C3-6cycloalkyl-C0-6alkyl halogen, CF3, ORf, S(O)kRf, CORf, NO2, N(Rf)2, CO(NRf)2, CH2N(Rf)2, or Rb and Rc are joined together to form a five or six membered aromatic or non-aromatic carbocyclic or heterocyclic ring, optionally substituted by up to three substituents chosen from halogen, CF3, C1-4alkyl, ORf, S(O)kRf, CORf, CO2Rf, OH, NO2, N(Rf)2, CO(NRf)2, and CH2N(Rf)2, or methylenedioxy;
or a pharmaceutically acceptable salt thereof.
or or or or wherein:
W is - (CHRg)b-V'- or phenyl;
A is a fibrinogen receptor antagonist template;
V' is CONR21 or NR21CO;
G is NRe, S or O;
Rg is H, C1-6alkyl, Het-C0-6alkyl, C3-7cycloalkyl-C0-6alkyl or Ar-C0-6alkyl;
R21 is Het-(CH2)0-6-U'-(CH2)1-6-, C3-7cycloalkyl-(CH2)0-6-U'-(CH2)1-6-, or Ar-(CH2)0-6-U'-(CH2)1-6-;
U' is CONRf or NRfCO;
Rf is H, C1-6alkyl or Ar-C1-6alkyl;
Re is H, C1-6alkyl, Ar-C1-6alkyl, Het-C1-6alkyl, C3-7cycloalkyl-C1-6alkyl, (CH2)qOH or (CH2)kCO2Rg;
k is 0, 1 or 2;
q is 1 or 2;
b is 0, 1 or 2;
Rb and Rc are independently selected from H, C1-6alkyl, Ar-C0-6alkyl, Het- C0-6alkyl, or C3-6cycloalkyl-C0-6alkyl halogen, CF3, ORf, S(O)kRf, CORf, NO2, N(Rf)2, CO(NRf)2, CH2N(Rf)2, or Rb and Rc are joined together to form a five or six membered aromatic or non-aromatic carbocyclic or heterocyclic ring, optionally substituted by up to three substituents chosen from halogen, CF3, C1-4alkyl, ORf, S(O)kRf, CORf, CO2Rf, OH, NO2, N(Rf)2, CO(NRf)2, and CH2N(Rf)2, or methylenedioxy;
or a pharmaceutically acceptable salt thereof.
2. A compound according to claim 1 wherein the fibrinogen receptor antagonist template A is wherein:
A1 to A5 form an accessible substituted seven-membered ring, which may be saturated or unsaturated, optionally containing up to two heteroatoms chosen from the group of O, S and N wherein S and N may be optionally oxidized;
D1 to D4 form an accessible substituted six membered ring, optionally containing up to two nitrogen atoms;
R is at least one substituent chosen from the group of R7, or Q-C1-4alkyl, Q-C2-4alkenyl, Q-C2-4alkynyl, optionally substituted by one or more of =O, R11 or R7;
R* is H, Q-C1-6alkyl, Q-C1-6oxoalkyl, Q-C2-6alkenyl, Q-C3-4oxoalkenyl, Q-C3-4oxoalkynyl, Q-C2-4alkynyl, C3-6cycloalkyl, Ar or Het, optionally substituted by one or more of R11;
Q is H, C3-6cycloalkyl, Het or Ar;
R7 is -COR8, -COCR'2R9, -C(S)R8, -S(O)mOR', -S(O)mNR'R", -PO(OR'), -PO(OR')2, -B(OR')2, -NO2 and Tet;
R8 is -OR', -NR'R", -NR'SO2R', -NR'OR', -OCR'2C(O)OR', -OCR'2OC(O)-R', -OCR'2C(O)NR'2, CF3 or AA;
R9 is -OR', -CN, -S(O)rR', S(O)mNR'2, -C(O)R'C(O)NR'2 or -CO2R';
R11 is H, halo, -OR12, -CN, -NR'R12, -NO2, -CF3, CF3S(O)r-, -CO2R', -CONR'2, Q-C0-6alkyl-, Q-C1-6oxoalkyl-, Q-C2-6alkenyl-, Q-C2-6alkynyl-, Q-C0-6alkyloxy-, Q-C0-6alkylamino- or Q-C0-6alkyl-S(O)r-;
R12 is R', -C(O)R', -C(O)NR'2, -C(O)OR15, -S(O)mR' or S(O)mNR'2;
R13 is R', -CF3, -SR', or -OR';
R14 is R', C(O)R', CN, NO2, SO2R' or C(O)OR15;
R15 is H, C1-6alkyl or Ar-C0-4alkyl;
R' is H, C1-6alkyl, C3-7cycloalkyl-C0-4alkyl or Ar-C0-4alkyl;
R" is R', -C(O)R' or -C(O)OR15;
R"' is R" or AA2;
AA1 is an amino acid attached through its amino group and having its carboxyl group optionally protected, and AA2 is an amino acid attached through its carboxyl group, and having its amino group optionally protected;
m is 1 or 2;
n is 0 to 3;
p is 0 or 1; and t is 0 to 2; or pharmaceutically acceptable salts thereof.
A1 to A5 form an accessible substituted seven-membered ring, which may be saturated or unsaturated, optionally containing up to two heteroatoms chosen from the group of O, S and N wherein S and N may be optionally oxidized;
D1 to D4 form an accessible substituted six membered ring, optionally containing up to two nitrogen atoms;
R is at least one substituent chosen from the group of R7, or Q-C1-4alkyl, Q-C2-4alkenyl, Q-C2-4alkynyl, optionally substituted by one or more of =O, R11 or R7;
R* is H, Q-C1-6alkyl, Q-C1-6oxoalkyl, Q-C2-6alkenyl, Q-C3-4oxoalkenyl, Q-C3-4oxoalkynyl, Q-C2-4alkynyl, C3-6cycloalkyl, Ar or Het, optionally substituted by one or more of R11;
Q is H, C3-6cycloalkyl, Het or Ar;
R7 is -COR8, -COCR'2R9, -C(S)R8, -S(O)mOR', -S(O)mNR'R", -PO(OR'), -PO(OR')2, -B(OR')2, -NO2 and Tet;
R8 is -OR', -NR'R", -NR'SO2R', -NR'OR', -OCR'2C(O)OR', -OCR'2OC(O)-R', -OCR'2C(O)NR'2, CF3 or AA;
R9 is -OR', -CN, -S(O)rR', S(O)mNR'2, -C(O)R'C(O)NR'2 or -CO2R';
R11 is H, halo, -OR12, -CN, -NR'R12, -NO2, -CF3, CF3S(O)r-, -CO2R', -CONR'2, Q-C0-6alkyl-, Q-C1-6oxoalkyl-, Q-C2-6alkenyl-, Q-C2-6alkynyl-, Q-C0-6alkyloxy-, Q-C0-6alkylamino- or Q-C0-6alkyl-S(O)r-;
R12 is R', -C(O)R', -C(O)NR'2, -C(O)OR15, -S(O)mR' or S(O)mNR'2;
R13 is R', -CF3, -SR', or -OR';
R14 is R', C(O)R', CN, NO2, SO2R' or C(O)OR15;
R15 is H, C1-6alkyl or Ar-C0-4alkyl;
R' is H, C1-6alkyl, C3-7cycloalkyl-C0-4alkyl or Ar-C0-4alkyl;
R" is R', -C(O)R' or -C(O)OR15;
R"' is R" or AA2;
AA1 is an amino acid attached through its amino group and having its carboxyl group optionally protected, and AA2 is an amino acid attached through its carboxyl group, and having its amino group optionally protected;
m is 1 or 2;
n is 0 to 3;
p is 0 or 1; and t is 0 to 2; or pharmaceutically acceptable salts thereof.
3. A compound according to claim 2 wherein:
A1 is CR1R1', CR1, NR1, N, O or S(O)x;
A2 is CR2R2', CR2, NR2;
A3 is CR3R3', CR3, NR3, N, O or S(O)x;
A4 is CR4R4', CR4, NR4, or N;
A5 is CR5R5', CR5, NR5, N, O or S(O)x;
D1-D4 are CH or N;
R1 and R1 are R* or R, or together are =O;
R2 and R2' are R*, R or =O;
R3 and R3' are R*, R or =O;
R4 and R4' are R*, R or =O;
R5 and R5' are R*, R or =O; and x is 0, 1 or 2.
A1 is CR1R1', CR1, NR1, N, O or S(O)x;
A2 is CR2R2', CR2, NR2;
A3 is CR3R3', CR3, NR3, N, O or S(O)x;
A4 is CR4R4', CR4, NR4, or N;
A5 is CR5R5', CR5, NR5, N, O or S(O)x;
D1-D4 are CH or N;
R1 and R1 are R* or R, or together are =O;
R2 and R2' are R*, R or =O;
R3 and R3' are R*, R or =O;
R4 and R4' are R*, R or =O;
R5 and R5' are R*, R or =O; and x is 0, 1 or 2.
4. A compound according to claim 2 wherein:
A1 is CR1R1, CR1, NR1, N, O or S; A2 is CR2R2', NR2 or CR2; A3 is CR3R3'; A4 is CR4R4, CR4, NR4, or N; A5 is CR5R5', CR5, NR5, N, O; D1 and D4 are CH; D2 or D3 is CH6; R2 or R4 are R; R3,R3' and R5,R5' are =O or R*,H.
A1 is CR1R1, CR1, NR1, N, O or S; A2 is CR2R2', NR2 or CR2; A3 is CR3R3'; A4 is CR4R4, CR4, NR4, or N; A5 is CR5R5', CR5, NR5, N, O; D1 and D4 are CH; D2 or D3 is CH6; R2 or R4 are R; R3,R3' and R5,R5' are =O or R*,H.
5. A compound according to claim 2 wherein:
A1 is CHR1, CR1, NR", N or S; A2 is CR2 or CR2R2'; A3 is CR3R3'; A4 is CR4R4' or NR4; A5 is CR5R5' D1-D4 are CH.
A1 is CHR1, CR1, NR", N or S; A2 is CR2 or CR2R2'; A3 is CR3R3'; A4 is CR4R4' or NR4; A5 is CR5R5' D1-D4 are CH.
6. A compound according to claim 2 wherein:
A1 is CR1, A2 is CR2, A3 is C=O, A4 is NR4 and A5 are CHR5.
A1 is CR1, A2 is CR2, A3 is C=O, A4 is NR4 and A5 are CHR5.
7. A compound according to claim 2 wherein:
A1 is NR1, A2 is CHCR2, A3 is CR3R3', A4 is NR4, and A5 are C=O.
A1 is NR1, A2 is CHCR2, A3 is CR3R3', A4 is NR4, and A5 are C=O.
8. A compound according to claim 2 wherein:
A1 and A4 are C=O, A2 is NR2, A3 is CHR3 and A5 is NR5.
A1 and A4 are C=O, A2 is NR2, A3 is CHR3 and A5 is NR5.
9. A compound according to claim 2 wherein:
A1 is NR1, A2 is CHR2, A3 is C=O, A4 is NR' and A5 is CHR5.
A1 is NR1, A2 is CHR2, A3 is C=O, A4 is NR' and A5 is CHR5.
10. A compound according to claim 2 wherein:
, , , , , , , , .
or
, , , , , , , , .
or
11. A compound according to claim 2 wherein:
.
.
12. A compound according to claim 11 wherein:
R' is H or C1-4alkyl; R2,R2' are H,-CH2CO2H; and R5R5' are H,H.
R' is H or C1-4alkyl; R2,R2' are H,-CH2CO2H; and R5R5' are H,H.
13. A compound according to formula (XXI) or (XXII):
or wherein:
B is -(CHRg)a-U-(CHRg)b-V- or phenyl or ;
A is a fibrinogen receptor antagonist template;
U and V are absent or CO, CRg2, C(=CRg2), S(O)k, O, NRg, CRgORg, CRg(ORk)CRg2, CRg2CRg(ORk), C(O)CRg2, CRg2C(O), CON Ri N Ri CO
OC(O), C(O)O, C(S)O, OC(S), C(S)NRg, NRgC(S), S(O)2NRg, NRgS(O)2 N=N, NRgNRg, NRgCRg2, NRgCRg2, CRg2O, OCRg2, C~C or CRg=CRg;
G is NRe, S or O;
Rg is H, C1-6alkyl, Het-C0-6alkyl, C3-7cycloalkyl-C0-6alkyl or Ar- C0-6alkyl;
Rk is Rg, -C(O)Rg, or -C(O)ORf;
Ri is is H, C1-6alkyl, Het-C0-6alkyl, C3-7cycloalkyl-C0-6alkyl, Ar- C0-6alkyl, Het-(CH2)0-6-U'-(cH2)1-6-, C3-7cycloalkyl-(CH2)0-6-U'-(CH2)1-6-, or Ar-(CH2)0-6-U'-(CH2)1-6-or C1-6alkyl substituted by one to three groups chosen from halogen, CN, NRg2, ORg, SRg, CO2Rg, and CON(Rg)2;
Rf is H, C1-6alkyl or Ar-C1-6alkyl;
Re is H, C1-6alkyl, Ar-C1-6alkyl, Het-C1-6alkyl, C3-7cycloalkyl-C1-6alkyl, (CH2)qOH or (CH2)kCO2Rg;
U' is CONRf or NRfCO, k is 0, 1 or 2;
q is 1 or 2;
a is 0, 1 or 2;
b is 0, 1 or 2;
Rb and Rc are independently selected from H, C1-6alkyl, Ar-C0-6alkyl, Het- C0-6alkyl, or C3-6cycloalkyl-C0-6alkyl, halogen, CF3, ORf, S(O)kRf, CORf, NO2, N(Rf)2, CO(NRf)2, CH2N(Rf)2, or Rb and Rc are joined together to form a five or six membered aromatic or non-aromatic carbocyclic or heterocyclic ring, optionally substituted by up to three substituents chosen from halogen, CF3, C1-4alkyl, ORf, S(O)kRf, CORf, CO2Rf OH, NO2, N(Rf)2, CO(NRf)2, and CH2N(Rf)2; or methylenedioxy;
or pharmaceutically acceptable salts thereof.
or wherein:
B is -(CHRg)a-U-(CHRg)b-V- or phenyl or ;
A is a fibrinogen receptor antagonist template;
U and V are absent or CO, CRg2, C(=CRg2), S(O)k, O, NRg, CRgORg, CRg(ORk)CRg2, CRg2CRg(ORk), C(O)CRg2, CRg2C(O), CON Ri N Ri CO
OC(O), C(O)O, C(S)O, OC(S), C(S)NRg, NRgC(S), S(O)2NRg, NRgS(O)2 N=N, NRgNRg, NRgCRg2, NRgCRg2, CRg2O, OCRg2, C~C or CRg=CRg;
G is NRe, S or O;
Rg is H, C1-6alkyl, Het-C0-6alkyl, C3-7cycloalkyl-C0-6alkyl or Ar- C0-6alkyl;
Rk is Rg, -C(O)Rg, or -C(O)ORf;
Ri is is H, C1-6alkyl, Het-C0-6alkyl, C3-7cycloalkyl-C0-6alkyl, Ar- C0-6alkyl, Het-(CH2)0-6-U'-(cH2)1-6-, C3-7cycloalkyl-(CH2)0-6-U'-(CH2)1-6-, or Ar-(CH2)0-6-U'-(CH2)1-6-or C1-6alkyl substituted by one to three groups chosen from halogen, CN, NRg2, ORg, SRg, CO2Rg, and CON(Rg)2;
Rf is H, C1-6alkyl or Ar-C1-6alkyl;
Re is H, C1-6alkyl, Ar-C1-6alkyl, Het-C1-6alkyl, C3-7cycloalkyl-C1-6alkyl, (CH2)qOH or (CH2)kCO2Rg;
U' is CONRf or NRfCO, k is 0, 1 or 2;
q is 1 or 2;
a is 0, 1 or 2;
b is 0, 1 or 2;
Rb and Rc are independently selected from H, C1-6alkyl, Ar-C0-6alkyl, Het- C0-6alkyl, or C3-6cycloalkyl-C0-6alkyl, halogen, CF3, ORf, S(O)kRf, CORf, NO2, N(Rf)2, CO(NRf)2, CH2N(Rf)2, or Rb and Rc are joined together to form a five or six membered aromatic or non-aromatic carbocyclic or heterocyclic ring, optionally substituted by up to three substituents chosen from halogen, CF3, C1-4alkyl, ORf, S(O)kRf, CORf, CO2Rf OH, NO2, N(Rf)2, CO(NRf)2, and CH2N(Rf)2; or methylenedioxy;
or pharmaceutically acceptable salts thereof.
14. A compound according to claim 13 wherein the fibrinogen receptor antagonist template A is wherein:
A1 to A5 form an accessible substituted seven-membered ring, which may be saturated or unsaturated, optionally containing up to two heteroatoms chosen from the group of O, S and N wherein S and N may be optionally oxidized;
D1 to D4 form an accessible substituted six membered ring, optionally containing up to two nitrogen atoms;
R is at least one substituent chosen from the group of R7, or Q-C1-4alkyl, Q-C2-4alkenyl, Q-C2-4alkynyl, optionally substituted by one or more of =O, R11 or R7;
R* is H, Q-C1-6alkyl, Q-C1-6oxoalkyl, Q-C2-6alkenyl, Q-C3-4oxoalkenyl, Q-C3-4oxoalkynyl, Q-C2-4alkynyl, C3-6cycloalkyl, Ar or Het, optionally substituted by one or more of R11;
Q is H, C3-6cycloalkyl, Het or Ar;
R7 is -COR8, -COCR'2R9, -C(S)R8, -S(O)mOR', -S(O)mNR'R", -PO(OR'), -PO(OR')2, -B(OR')2, -NO2 and Tet;
R8 is -OR', -NR'R", -NR'SO2R', -NR'OR', -OCR'2C(O)OR', -OCR'2OC(O)-R', -OCR'2C(O)NR'2, CF3 or AA;
R9 is -OR', -CN, -S(O)rR', S(O)mNR'2, -C(O)R'C(O)NR'2 or -CO2R';
R11 is H, halo, -OR12, -CN, -NR'R12, -NO2, -CF3, CF3S(O)r, -CO2R', -CONR'2, Q-C0-6alkyl-, Q-C1-6oxoalkyl-, Q-C2-6alkenyl-, Q-C2-6alkynyl-, Q-C0-6alkyloxy-, Q-C0-6alkylamino- or Q-C0-6alkyl-S(O),-;
R12 is R', -C(O)R', -C(O)NR'2, -C(O)OR15, -S(O)mR' or S(O)mNR'2;
R13 is R', -CF3, -SR', or -OR';
R14 is R', C(O)R', CN, NO2, SO2R' or C(O)OR15;
R15 is H, C1-6alkyl or Ar-C0-4alkyl;
R' is H, C1-6alkyl, C3-7cycloalkyl-C0-4alkyl or Ar-C0-4alkyl;
R" is R', -C(O)R' or -C(O)OR15;
R''' is R" or AA2;
AA1 is an amino acid attached through its amino group and having its carboxyl group optionally protected, and AA2 is an amino acid attached through its carboxyl group, and having its amino group optionally protected;
m is 1 or 2;
n is 0 to 3;
p is 0 or 1; and t is 0 to 2; or pharmaceutically acceptable salts thereof.
A1 to A5 form an accessible substituted seven-membered ring, which may be saturated or unsaturated, optionally containing up to two heteroatoms chosen from the group of O, S and N wherein S and N may be optionally oxidized;
D1 to D4 form an accessible substituted six membered ring, optionally containing up to two nitrogen atoms;
R is at least one substituent chosen from the group of R7, or Q-C1-4alkyl, Q-C2-4alkenyl, Q-C2-4alkynyl, optionally substituted by one or more of =O, R11 or R7;
R* is H, Q-C1-6alkyl, Q-C1-6oxoalkyl, Q-C2-6alkenyl, Q-C3-4oxoalkenyl, Q-C3-4oxoalkynyl, Q-C2-4alkynyl, C3-6cycloalkyl, Ar or Het, optionally substituted by one or more of R11;
Q is H, C3-6cycloalkyl, Het or Ar;
R7 is -COR8, -COCR'2R9, -C(S)R8, -S(O)mOR', -S(O)mNR'R", -PO(OR'), -PO(OR')2, -B(OR')2, -NO2 and Tet;
R8 is -OR', -NR'R", -NR'SO2R', -NR'OR', -OCR'2C(O)OR', -OCR'2OC(O)-R', -OCR'2C(O)NR'2, CF3 or AA;
R9 is -OR', -CN, -S(O)rR', S(O)mNR'2, -C(O)R'C(O)NR'2 or -CO2R';
R11 is H, halo, -OR12, -CN, -NR'R12, -NO2, -CF3, CF3S(O)r, -CO2R', -CONR'2, Q-C0-6alkyl-, Q-C1-6oxoalkyl-, Q-C2-6alkenyl-, Q-C2-6alkynyl-, Q-C0-6alkyloxy-, Q-C0-6alkylamino- or Q-C0-6alkyl-S(O),-;
R12 is R', -C(O)R', -C(O)NR'2, -C(O)OR15, -S(O)mR' or S(O)mNR'2;
R13 is R', -CF3, -SR', or -OR';
R14 is R', C(O)R', CN, NO2, SO2R' or C(O)OR15;
R15 is H, C1-6alkyl or Ar-C0-4alkyl;
R' is H, C1-6alkyl, C3-7cycloalkyl-C0-4alkyl or Ar-C0-4alkyl;
R" is R', -C(O)R' or -C(O)OR15;
R''' is R" or AA2;
AA1 is an amino acid attached through its amino group and having its carboxyl group optionally protected, and AA2 is an amino acid attached through its carboxyl group, and having its amino group optionally protected;
m is 1 or 2;
n is 0 to 3;
p is 0 or 1; and t is 0 to 2; or pharmaceutically acceptable salts thereof.
15. A compound according to claim 14 wherein:
A1 is CR1R1', CR1, NR1, N, O or S(O)x;
A2 is CR2R2', CR2, NR2;
A3 is CR3R3', CR3, NR3, N, O or S(O)x;
A4 is CR4R4', CR4, NR4, or N;
A5 is CR5R5', CR5, NR5, N, O or S(O)x;
D1-D4 are CH or N;
R1 and R1' are R* or R, or together are =O;
R2 and R2' are R*, R or =O;
R3 and R3' are R*, R or =O;
R4 and R4' are R*, R or =O;
R5 and R5' are R*, R or =O; and x is 0, 1 or 2.
A1 is CR1R1', CR1, NR1, N, O or S(O)x;
A2 is CR2R2', CR2, NR2;
A3 is CR3R3', CR3, NR3, N, O or S(O)x;
A4 is CR4R4', CR4, NR4, or N;
A5 is CR5R5', CR5, NR5, N, O or S(O)x;
D1-D4 are CH or N;
R1 and R1' are R* or R, or together are =O;
R2 and R2' are R*, R or =O;
R3 and R3' are R*, R or =O;
R4 and R4' are R*, R or =O;
R5 and R5' are R*, R or =O; and x is 0, 1 or 2.
16. A compound according to claim 14 wherein:
A1 is CR1R1', CR1, NR1, N, O or S; A2 is CR2R2', NR2 or CR2; A3 is CR3R3'; A4 is CR4R4', CR4, NR4, or N; A5 is CR5R5', CR5, NR5, N, O; D1l and D4 are CH; D2 or D3 is CH6; R2 or R4 are R; R3,R3' and R5,R5' are =O or R*,H.
A1 is CR1R1', CR1, NR1, N, O or S; A2 is CR2R2', NR2 or CR2; A3 is CR3R3'; A4 is CR4R4', CR4, NR4, or N; A5 is CR5R5', CR5, NR5, N, O; D1l and D4 are CH; D2 or D3 is CH6; R2 or R4 are R; R3,R3' and R5,R5' are =O or R*,H.
17. A compound according to claim 14 wherein:
A1 is CHR1, CR1, NR", N or S; A2 is CR2 or CR2R2'; A3 is CR3R3'; A4 is CR4R4' or NR4; A5 is CR5R5' D1-D4 are CH.
A1 is CHR1, CR1, NR", N or S; A2 is CR2 or CR2R2'; A3 is CR3R3'; A4 is CR4R4' or NR4; A5 is CR5R5' D1-D4 are CH.
18. A compound according to claim 14 wherein:
A1 is CR1, A2 is CR2, A3 is C=O, A4 is NR4 and A5 are CHR5.
A1 is CR1, A2 is CR2, A3 is C=O, A4 is NR4 and A5 are CHR5.
19. A compound according to claim 14 wherein:
A1 is NR1, A2 is CHCR2, A3 is CR3R3', A4 is NR4, and A5 are C=O.
A1 is NR1, A2 is CHCR2, A3 is CR3R3', A4 is NR4, and A5 are C=O.
20. A compound according to claim 14 wherein:
A1 and A4 are C=O, A2 is NR2, A3 is CHR3 and A5 is NR5.
A1 and A4 are C=O, A2 is NR2, A3 is CHR3 and A5 is NR5.
21. A compound according to claim 14 wherein:
A1 is NR1, A2 is CHR2, A3 is C=O, A4 is NR' and As is CHR5.
A1 is NR1, A2 is CHR2, A3 is C=O, A4 is NR' and As is CHR5.
22. A compound according to claim 14 wherein:
, , , , , , , , or ,
, , , , , , , , or ,
23. A compound according to claim 14 wherein:
24. A compound according to claim 23 wherein:
R1 is H or C1-4alkyl; R2, R2' are H,-CH2CO2H; and R5R5' are H,H.
R1 is H or C1-4alkyl; R2, R2' are H,-CH2CO2H; and R5R5' are H,H.
25. A compound selected from the group of:
5-[[[(Benzimidazol-2-yl)methyl]methylamino]carbonyl]-1H-benzimidazol-2-aminoacetic acid;
(~)-2,3,4,5-Tetrahydro-7-[[[(benzimidazol-2-yl)methyl]-methylamino]carbonyl]-4-(3,3-dimethylbutyl)-3-oxo-1H-1,4-benzodiazepine-2-acetic acid;
(S)-2,3,4,5-Tetrahydro-4-methyl-3-oxo-7-[[[(5-trifluoromethylbenzimidazol-2-yl)methyl]methylamino]carbonyl]-1H-1,4-benzodiazepine-2-acetic acid;
(S)-2,3,4,5-Tetrahydro-7-[[[(4,7-dimethoxybenzimidazol-2-yl)methyl]methylamino]carbonyl]-4-methyl-3-oxo-1H-1,4-benzodiazepine-2-acetic acid;
(~)-2,3,4,5-Tetrahydro-7-[[[(benzimidazol-2-yl)methyl]amino]carbonyl]-4-(3,3-dimethylbutyl)-3-oxo-1H-1,4-benzodiazepine-2-acetic acid;
(S)-2,3,4,5-Tetrahydro-7-[[[(4-methylbenzimidazol-2-yl)methyl]methylamino]carbony]-4-methyl-3-oxo-1H-1,4-benzodiazepine-2-acetic acid;
S)-2,3,4,5-Tetrahydro-7-[[N-[(benzimidazol-2-yl)methyl]-N-(4-aminobutyl)amino]carbonyl]4-methyl-3-oxo-1H-1,4-benzodiazepine-2-acetic acid;
(S)-2,3,4,5-Tetrahydro-7-[[N-(benzimidazol-2-yl)methyl-N-(2-cyanomethyl)amino]carbonyl]-4-methyl-3-oxo-1H-1,4-benzodiazepine-2-actic acid;
(S)-2,3,4,5-Tetrahydro-7-[[[(benzimidazol-2-yl)methyl]amino]carbonyl]-3-oxo-4-(4-phthalimidobutyl)-1H-1,4-benzodiazepine-2-acetic acid;
4-[[[3-(Benzimidazol-2-yl)propyl]amino]carbonyl]piperidine-1-acetic acid;
4-[[[3-(Benzimidazol-2-yl)propyl]amino]carbonyl]phenylacetic acid;
(S)-2,3,4,5-Tetrahydro-7-[[[4-aza-5,7-dimethylbenzimidazol-2-yl)methyl]methylamino]carbonyl]-4-methyl-3-oxo-1H-1,4-benzodiazepine-2-acetic acid;
(~)-2,3,4,5-Tetrahydro-7-[[[(benzimidazol-2-yl)methyl]methylamino]-carbonyl]-4-[2-(3,4-methylenedioxyphenyl)ethyl]-3-oxo-1H-1,4-benzodiazepine-2-acetic acid;
(~)-2,3,4,5-Tetrahydro-7-[[[(benzimidazol-2-yl)methyl]amino]carbonyl]-4-(2-methoxyethyl)-3-oxo-1H-1,4-benzodiazepine-2-acetic acid;
(S)-2,3,4,5-Tetrahydro-7-[[[(benzimidazol-2-yl)methyl]methylamino]-carbonyl]-4-methyl-3-oxo-1H-1,4-benzodiazepine-2-acetamide;
(~)-2,3,4,5-Tetrahydro-7-[[[[1-[(benzimidazol-2-yl)methyl]benzimidazol-2-yl]methyl]amino]carbonyl]-4-methyl-3-oxo-1H-1,4-benzodiazepine-2-acetic acid;
(S)-2,3,4,5-Tetrahydro-7-[[[(benzimidazol-2-yl)methyl]methylamino]carbonyl]-3-oxo-1H-1,4-benzodiazepine-2-acetic acid;
(S)-2,3,4,5-Tetrahydro-7-[[bis[(benzimidazol-2-yl)methyl]amino]carbonyl]-4-methyl-3-oxo-1H-1,4-benzodiazepine-2-acetic acid;
(~)-2,3,4,5-Tetrahydro-7-[[[(4-azabenzimidazol-2-yl)methyl]methylamino]carbonyl]-4-(3,3-dimethylbutyl)-3-oxo-1H-1,4-benzodiazepine-2-acetic acid;
(~)-2,3,4,5-Tetrahydro-7-[[[(benzimidazol-2-yl)methyl]methylamino]carbonyl]-3-oxo-4-(2,2,2-trifluoroethyl)-1H-1,4-benzodiazepine-2-acetic acid;
(+)-2,3,4,5-Tetrahydro-7-[[2-(benzimidazol-2-yl)acetyl]amino]-5-oxo-4-(2-phenylethyl)-1H-1,4-benzodiazepine-2-acetic acid;
(+)-2,3,4,5-Tetrahydro-7-[[[(benzimidazol-2-yl)methyl]amino]carbonyl]-3-oxo-4-(2,2,2-trifluoroethyl)-1H-1,4-benzodiazepine-2-acetic acid;
(S)-2,3,4,5-Tetrahydro-7-[[[(5,6-difluorobenzimidazol-2-yl)methyl]methylamino]carbonyl]-4-methyl-3-oxo-1H-1,4-benzodiazepine-2-acetic acid;
(~)-2,3,4,5-Tetrahydro-7-[[bis[(benzimidazol-2-yl)methyl]amino]carbonyl]-3-oxo-4-(2-phenylethyl)-1H-1,4-benzodiazepine-2-acetic acid;
(S)-2,3,4,5-Tetrahydro-7-[[[(4-aza-5-methylbenzimidazol-2-yl)methyl]amino]carbonyl]-4-methyl-3-oxo-1H-1,4-benzodiazepine-2-acetic acid;
(S)-2,3,4,5-Tetrahydro-4-methyl-7-[[[(4-nitrobenzimidazol-2-yl)methyl]methylamino]carbonyl]-3-oxo-1H-1,4-benzodiazepine-2-acetic acid;
(~)-2,3,4,5-Tetrahydro-7-[[[(4-aza-5-methylbenzimidazol-2-yl)methyl]amino]carbonyl]-3-oxo-4-(2,2,2-trifluoroethyl)-1H-1,4-benzodiazepine-2-acetic acid;
(~)-4-[4-[[[(1H-Benzimidazol-2-yl)methyl]methylamino]carbonyl]phenyl]-3-phenylbutanoic acid;
(~)-3-[[[4-(4-Azabenzimidazol-2-yl)butanoyl]glycyl]amino]4-pentynoic acid;
(S)-2,3,4,5-Tetrahydro-7-[[[[1-(2-hydroxyethyl)benzimidazol-2-yl]methyl]amino]carbonyl]-4-methyl-3-oxo-1H-1,4-benzodiazepine-2-acetic acid;
(~)-2,3,4,5-Tetrahydro-7-[[[(4-aza-5-methylbenzimidazol-2-yl)methyl]amino]carbonyl]-4-(2-methoxyethyl)-3-oxo-1H-1,4-benzodiazepine-2-acetic acid;
(S)-2,3,4,5-Tetrahydro-7-[[[(4-aminobenzimidazol-2-yl)methyl]methylamino]carbonyl]-4-methyl-3-oxo-1H-1,4-benzodiazepine-2-acetic acid;
Ethyl (S)-2,3,4,5-tetrahydro-7-[[[(benzimidazol-2-yl)methyl]methylamino]carbonyl]-4-methyl-3-oxo-1H-1,4-benzodiazepine-2-acetate;
(S)-2,3,4,5-Tetrahydro-7-[[[(benzimidazol-2-yl)methyl]methylamino]carbonyl]-4-methyl-3-oxo-1H-1,4-benzodiazepine-2-acetic acid, [(2,2-dimethyl-2-methoxyacetyl)oxy]methyl ester;
2,3,4,5-Tetrahydro-7-[[[(1R)-(benzimidazol-2-yl)ethyl]methylamino]carbonyl]-4-methyl-3-oxo-1H-1,4-benzodiazepine-(2S)-acetic acid;
(~)-N-[2-(Aminomethyl)4-[[[(4-aza-5-methylbenzimidazol-2-yl)methyl]methylamino]carbonyl]phenyl]aspartic acid;
(~)-2,3,4,5-Tetrahydro-4-methyl-3-oxo-7-[[[(phenanthrimiidazol-2-yl)methyl]amino]carbonyl]- 1 H- 1 ,4-benzodiazepine-2-acetic acid;
(~)-2,3,4,5-Tetrahydro-7-[3-(benzimidazol-2-yl)phenyl]-4-methyl-3-oxo-1H-1,4-benzodiazepine-2-acetic acid;
(~)-4-[4-[[[(Benzimidazol-2-yl)methyl]methylamino]carbonyl]phenyl]-3-(dimethylaminocarbonyl)butanoic acid;
(S)-2,3,4,5-Tetrahydro-7-[[[(benzimidazol-2-yl)methyl]amino]carbonyl]-3-oxo-4-[2-(pyrid-3-yl)ethyl]-1H-1,4-benzodiazepine-2-acetate;
(S)-2,3,4,5-Tetrahydro-7-[[[(4-aza-5-methylbenzimidazol-2-yl)methyl]methylamino]carbonyl]-3-oxo-1H-1,4-benzodiazepine-2-acetic acid;
~)-2,3,4,5-Tetrahydro-7-[[N-[benzimidazol-2-yl)methyl]-N-[[4-(2-carboxybenzoyl)amino]butyl]amino]carbonyl]-3-oxo-4-(2-phenylethyl)-1H-1,4-benzodiazepine-2-acetic acid;
(~)-2,3,4,5-Tetrahydro-7-[[N-[(benzimidazol-2-yl)methyl]-N-[[4-(4-azido-2-hydroxybenzoyl)amino]butyl]amino]carbonyl]-3-oxo-4-(2-phenylethyl)-1H-1,4-benzodiazepine-2-acetic acid;
Ethyl (S)-2,3,4,5-tetrahydro-7-[[[(4-aza-5-methylbenzimidazol-2-yl)methyl]amino]carbonyl]4-methyl-3-oxo-1H-1,4-benzodiazepine-2-acetate;
2,3,4,5-Tetrahydro-7-[[N-[(benzimidazol-2-yl)methyl]-N-[[[(+)-biotinoyl]amino]butyl]amino]carbonyl]-3-oxo-4-(2-phenylethyl)-1H-1,4-benzodiazepine-(2RS)-acetic acid;
2,3,4,5-Tetrahydro-7-[[[(1S)-(benzimidazol-2-yl)ethyl]methylamino]carbonyl]-4-methyl-3-oxo-1H-1,4-benzodiazepine-(2S)-acetic acid;
(S)-2,3,4,5-Tetrahydro-7-[[[(imidazo(1,2a)pyrid-2-yl)methyl]methylamino]carbonyl]4-methyl-3-oxo-1H-1,4-benzodiazepine-2-acetic acid;
(S)-2,3,4,5-Tetrahydro-7-[[[(benzimidazol-2-yl)methyl]amino]carbonyl]-3-oxo-1H-1,4-benzodiazepine-2-acetic acid;
(~)-5-[[2,3,4,5-Tetrahydro-7-[[[(benzimidazol-2-yl)methyl]amino]carbonyl]-3-oxo-4-(2-phenylethyl)-1H-1,4-benzodiazepin-2-yl]methyl]tetrazole;
(S)-2,3,4,5-Tetrahydro-7-[[[(4-aza-5-methylbenzimidazol-2-yl)methyl]amino]carbonyl]-3-oxo-1H-1,4-benzodiazepine-2-acetic acid;
(~)-2,3,4,5-Tetrahydro-7-[3-(benzimidazol-2-yl)propyl]-4-methyl-3-oxo-1H-1,4-benzodiazepine-2-acetic acid;
(~)-2,3,4,5-Tetrahydro-7-[[N-[(benzimidazol-2-yl)methyl]-N-(4-aminobutyl)amino]carbonyl]-3-oxo-4-(2-phenylethyl)-1H-1,4-benzodiazepine-2-acetic acid;
(~)-2,3,4,5-Tetrahydro-7-[[[(benzimidazol-2-yl)methyl]amino]carbonyl]-4-methyl-3-oxo-1H-1,4-benzodiazepine-2-(N-hydroxy)acetamide;
Ethyl (~)-3-[[[2-(Benzimidazol-2-yl)ethyl]amino]succinoyl]amino-4-pentynoate;
(~)-3-[[[2-(Benzimidazol-2-yl)ethyl]amino]succinoyl]amino-4-pentynoic acid;
(~)-2,3,4,5-Tetrahydro-7-[[N-[(benzirnidazol-2-yl)methyl]-N-[[4-(4-azido-3-iodo-2-hydroxybenzoyl)amino]butyl]amino]carbonyl]-3-oxo-4-(2-phenylethyl)-1H-1,4-benzodiazepine-2-acetic acid;
2,3,4,5-Tetrahydro-7-[[[(1S)-(benzimidazol-2-yl)ethyl]amino]carbonyl]-4-methyl-3-oxo-1H-1,4-benzodiazepine-(2S)-acetic acid; and 2,3,4,5-Tetrahydro-7-[[[(1R)-(benzimidazol-2-yl)ethyl]amino]carbonyl]-4-methyl-3-oxo-1H-1,4-benzodiazepine-(2S)-acetic acid; and (~)-7-[[[(4,5-Dimethyl-1H-imidazol-2-yl)methyl]methylamino]carbonyl]-2,3,4,5-tetrahydro-4-methyl-3-oxo-1H-1,4-benzodiazepine-2-acetic acid;
or pharmaceutically acceptable salts thereof.
5-[[[(Benzimidazol-2-yl)methyl]methylamino]carbonyl]-1H-benzimidazol-2-aminoacetic acid;
(~)-2,3,4,5-Tetrahydro-7-[[[(benzimidazol-2-yl)methyl]-methylamino]carbonyl]-4-(3,3-dimethylbutyl)-3-oxo-1H-1,4-benzodiazepine-2-acetic acid;
(S)-2,3,4,5-Tetrahydro-4-methyl-3-oxo-7-[[[(5-trifluoromethylbenzimidazol-2-yl)methyl]methylamino]carbonyl]-1H-1,4-benzodiazepine-2-acetic acid;
(S)-2,3,4,5-Tetrahydro-7-[[[(4,7-dimethoxybenzimidazol-2-yl)methyl]methylamino]carbonyl]-4-methyl-3-oxo-1H-1,4-benzodiazepine-2-acetic acid;
(~)-2,3,4,5-Tetrahydro-7-[[[(benzimidazol-2-yl)methyl]amino]carbonyl]-4-(3,3-dimethylbutyl)-3-oxo-1H-1,4-benzodiazepine-2-acetic acid;
(S)-2,3,4,5-Tetrahydro-7-[[[(4-methylbenzimidazol-2-yl)methyl]methylamino]carbony]-4-methyl-3-oxo-1H-1,4-benzodiazepine-2-acetic acid;
S)-2,3,4,5-Tetrahydro-7-[[N-[(benzimidazol-2-yl)methyl]-N-(4-aminobutyl)amino]carbonyl]4-methyl-3-oxo-1H-1,4-benzodiazepine-2-acetic acid;
(S)-2,3,4,5-Tetrahydro-7-[[N-(benzimidazol-2-yl)methyl-N-(2-cyanomethyl)amino]carbonyl]-4-methyl-3-oxo-1H-1,4-benzodiazepine-2-actic acid;
(S)-2,3,4,5-Tetrahydro-7-[[[(benzimidazol-2-yl)methyl]amino]carbonyl]-3-oxo-4-(4-phthalimidobutyl)-1H-1,4-benzodiazepine-2-acetic acid;
4-[[[3-(Benzimidazol-2-yl)propyl]amino]carbonyl]piperidine-1-acetic acid;
4-[[[3-(Benzimidazol-2-yl)propyl]amino]carbonyl]phenylacetic acid;
(S)-2,3,4,5-Tetrahydro-7-[[[4-aza-5,7-dimethylbenzimidazol-2-yl)methyl]methylamino]carbonyl]-4-methyl-3-oxo-1H-1,4-benzodiazepine-2-acetic acid;
(~)-2,3,4,5-Tetrahydro-7-[[[(benzimidazol-2-yl)methyl]methylamino]-carbonyl]-4-[2-(3,4-methylenedioxyphenyl)ethyl]-3-oxo-1H-1,4-benzodiazepine-2-acetic acid;
(~)-2,3,4,5-Tetrahydro-7-[[[(benzimidazol-2-yl)methyl]amino]carbonyl]-4-(2-methoxyethyl)-3-oxo-1H-1,4-benzodiazepine-2-acetic acid;
(S)-2,3,4,5-Tetrahydro-7-[[[(benzimidazol-2-yl)methyl]methylamino]-carbonyl]-4-methyl-3-oxo-1H-1,4-benzodiazepine-2-acetamide;
(~)-2,3,4,5-Tetrahydro-7-[[[[1-[(benzimidazol-2-yl)methyl]benzimidazol-2-yl]methyl]amino]carbonyl]-4-methyl-3-oxo-1H-1,4-benzodiazepine-2-acetic acid;
(S)-2,3,4,5-Tetrahydro-7-[[[(benzimidazol-2-yl)methyl]methylamino]carbonyl]-3-oxo-1H-1,4-benzodiazepine-2-acetic acid;
(S)-2,3,4,5-Tetrahydro-7-[[bis[(benzimidazol-2-yl)methyl]amino]carbonyl]-4-methyl-3-oxo-1H-1,4-benzodiazepine-2-acetic acid;
(~)-2,3,4,5-Tetrahydro-7-[[[(4-azabenzimidazol-2-yl)methyl]methylamino]carbonyl]-4-(3,3-dimethylbutyl)-3-oxo-1H-1,4-benzodiazepine-2-acetic acid;
(~)-2,3,4,5-Tetrahydro-7-[[[(benzimidazol-2-yl)methyl]methylamino]carbonyl]-3-oxo-4-(2,2,2-trifluoroethyl)-1H-1,4-benzodiazepine-2-acetic acid;
(+)-2,3,4,5-Tetrahydro-7-[[2-(benzimidazol-2-yl)acetyl]amino]-5-oxo-4-(2-phenylethyl)-1H-1,4-benzodiazepine-2-acetic acid;
(+)-2,3,4,5-Tetrahydro-7-[[[(benzimidazol-2-yl)methyl]amino]carbonyl]-3-oxo-4-(2,2,2-trifluoroethyl)-1H-1,4-benzodiazepine-2-acetic acid;
(S)-2,3,4,5-Tetrahydro-7-[[[(5,6-difluorobenzimidazol-2-yl)methyl]methylamino]carbonyl]-4-methyl-3-oxo-1H-1,4-benzodiazepine-2-acetic acid;
(~)-2,3,4,5-Tetrahydro-7-[[bis[(benzimidazol-2-yl)methyl]amino]carbonyl]-3-oxo-4-(2-phenylethyl)-1H-1,4-benzodiazepine-2-acetic acid;
(S)-2,3,4,5-Tetrahydro-7-[[[(4-aza-5-methylbenzimidazol-2-yl)methyl]amino]carbonyl]-4-methyl-3-oxo-1H-1,4-benzodiazepine-2-acetic acid;
(S)-2,3,4,5-Tetrahydro-4-methyl-7-[[[(4-nitrobenzimidazol-2-yl)methyl]methylamino]carbonyl]-3-oxo-1H-1,4-benzodiazepine-2-acetic acid;
(~)-2,3,4,5-Tetrahydro-7-[[[(4-aza-5-methylbenzimidazol-2-yl)methyl]amino]carbonyl]-3-oxo-4-(2,2,2-trifluoroethyl)-1H-1,4-benzodiazepine-2-acetic acid;
(~)-4-[4-[[[(1H-Benzimidazol-2-yl)methyl]methylamino]carbonyl]phenyl]-3-phenylbutanoic acid;
(~)-3-[[[4-(4-Azabenzimidazol-2-yl)butanoyl]glycyl]amino]4-pentynoic acid;
(S)-2,3,4,5-Tetrahydro-7-[[[[1-(2-hydroxyethyl)benzimidazol-2-yl]methyl]amino]carbonyl]-4-methyl-3-oxo-1H-1,4-benzodiazepine-2-acetic acid;
(~)-2,3,4,5-Tetrahydro-7-[[[(4-aza-5-methylbenzimidazol-2-yl)methyl]amino]carbonyl]-4-(2-methoxyethyl)-3-oxo-1H-1,4-benzodiazepine-2-acetic acid;
(S)-2,3,4,5-Tetrahydro-7-[[[(4-aminobenzimidazol-2-yl)methyl]methylamino]carbonyl]-4-methyl-3-oxo-1H-1,4-benzodiazepine-2-acetic acid;
Ethyl (S)-2,3,4,5-tetrahydro-7-[[[(benzimidazol-2-yl)methyl]methylamino]carbonyl]-4-methyl-3-oxo-1H-1,4-benzodiazepine-2-acetate;
(S)-2,3,4,5-Tetrahydro-7-[[[(benzimidazol-2-yl)methyl]methylamino]carbonyl]-4-methyl-3-oxo-1H-1,4-benzodiazepine-2-acetic acid, [(2,2-dimethyl-2-methoxyacetyl)oxy]methyl ester;
2,3,4,5-Tetrahydro-7-[[[(1R)-(benzimidazol-2-yl)ethyl]methylamino]carbonyl]-4-methyl-3-oxo-1H-1,4-benzodiazepine-(2S)-acetic acid;
(~)-N-[2-(Aminomethyl)4-[[[(4-aza-5-methylbenzimidazol-2-yl)methyl]methylamino]carbonyl]phenyl]aspartic acid;
(~)-2,3,4,5-Tetrahydro-4-methyl-3-oxo-7-[[[(phenanthrimiidazol-2-yl)methyl]amino]carbonyl]- 1 H- 1 ,4-benzodiazepine-2-acetic acid;
(~)-2,3,4,5-Tetrahydro-7-[3-(benzimidazol-2-yl)phenyl]-4-methyl-3-oxo-1H-1,4-benzodiazepine-2-acetic acid;
(~)-4-[4-[[[(Benzimidazol-2-yl)methyl]methylamino]carbonyl]phenyl]-3-(dimethylaminocarbonyl)butanoic acid;
(S)-2,3,4,5-Tetrahydro-7-[[[(benzimidazol-2-yl)methyl]amino]carbonyl]-3-oxo-4-[2-(pyrid-3-yl)ethyl]-1H-1,4-benzodiazepine-2-acetate;
(S)-2,3,4,5-Tetrahydro-7-[[[(4-aza-5-methylbenzimidazol-2-yl)methyl]methylamino]carbonyl]-3-oxo-1H-1,4-benzodiazepine-2-acetic acid;
~)-2,3,4,5-Tetrahydro-7-[[N-[benzimidazol-2-yl)methyl]-N-[[4-(2-carboxybenzoyl)amino]butyl]amino]carbonyl]-3-oxo-4-(2-phenylethyl)-1H-1,4-benzodiazepine-2-acetic acid;
(~)-2,3,4,5-Tetrahydro-7-[[N-[(benzimidazol-2-yl)methyl]-N-[[4-(4-azido-2-hydroxybenzoyl)amino]butyl]amino]carbonyl]-3-oxo-4-(2-phenylethyl)-1H-1,4-benzodiazepine-2-acetic acid;
Ethyl (S)-2,3,4,5-tetrahydro-7-[[[(4-aza-5-methylbenzimidazol-2-yl)methyl]amino]carbonyl]4-methyl-3-oxo-1H-1,4-benzodiazepine-2-acetate;
2,3,4,5-Tetrahydro-7-[[N-[(benzimidazol-2-yl)methyl]-N-[[[(+)-biotinoyl]amino]butyl]amino]carbonyl]-3-oxo-4-(2-phenylethyl)-1H-1,4-benzodiazepine-(2RS)-acetic acid;
2,3,4,5-Tetrahydro-7-[[[(1S)-(benzimidazol-2-yl)ethyl]methylamino]carbonyl]-4-methyl-3-oxo-1H-1,4-benzodiazepine-(2S)-acetic acid;
(S)-2,3,4,5-Tetrahydro-7-[[[(imidazo(1,2a)pyrid-2-yl)methyl]methylamino]carbonyl]4-methyl-3-oxo-1H-1,4-benzodiazepine-2-acetic acid;
(S)-2,3,4,5-Tetrahydro-7-[[[(benzimidazol-2-yl)methyl]amino]carbonyl]-3-oxo-1H-1,4-benzodiazepine-2-acetic acid;
(~)-5-[[2,3,4,5-Tetrahydro-7-[[[(benzimidazol-2-yl)methyl]amino]carbonyl]-3-oxo-4-(2-phenylethyl)-1H-1,4-benzodiazepin-2-yl]methyl]tetrazole;
(S)-2,3,4,5-Tetrahydro-7-[[[(4-aza-5-methylbenzimidazol-2-yl)methyl]amino]carbonyl]-3-oxo-1H-1,4-benzodiazepine-2-acetic acid;
(~)-2,3,4,5-Tetrahydro-7-[3-(benzimidazol-2-yl)propyl]-4-methyl-3-oxo-1H-1,4-benzodiazepine-2-acetic acid;
(~)-2,3,4,5-Tetrahydro-7-[[N-[(benzimidazol-2-yl)methyl]-N-(4-aminobutyl)amino]carbonyl]-3-oxo-4-(2-phenylethyl)-1H-1,4-benzodiazepine-2-acetic acid;
(~)-2,3,4,5-Tetrahydro-7-[[[(benzimidazol-2-yl)methyl]amino]carbonyl]-4-methyl-3-oxo-1H-1,4-benzodiazepine-2-(N-hydroxy)acetamide;
Ethyl (~)-3-[[[2-(Benzimidazol-2-yl)ethyl]amino]succinoyl]amino-4-pentynoate;
(~)-3-[[[2-(Benzimidazol-2-yl)ethyl]amino]succinoyl]amino-4-pentynoic acid;
(~)-2,3,4,5-Tetrahydro-7-[[N-[(benzirnidazol-2-yl)methyl]-N-[[4-(4-azido-3-iodo-2-hydroxybenzoyl)amino]butyl]amino]carbonyl]-3-oxo-4-(2-phenylethyl)-1H-1,4-benzodiazepine-2-acetic acid;
2,3,4,5-Tetrahydro-7-[[[(1S)-(benzimidazol-2-yl)ethyl]amino]carbonyl]-4-methyl-3-oxo-1H-1,4-benzodiazepine-(2S)-acetic acid; and 2,3,4,5-Tetrahydro-7-[[[(1R)-(benzimidazol-2-yl)ethyl]amino]carbonyl]-4-methyl-3-oxo-1H-1,4-benzodiazepine-(2S)-acetic acid; and (~)-7-[[[(4,5-Dimethyl-1H-imidazol-2-yl)methyl]methylamino]carbonyl]-2,3,4,5-tetrahydro-4-methyl-3-oxo-1H-1,4-benzodiazepine-2-acetic acid;
or pharmaceutically acceptable salts thereof.
26. A compound selected from the group of:
3-[[3-[2-(benzimidazol-2-yl)ethyl]isoxazolin-5(R,S)-yl]acetyl]amino-3(R,S)-melhylpropanoic acid;
3-{3,4-dihydro-8-[[[(benzimidazol-2-yl)methyl]methylamino]carbonyl]-1-methyl-2,5-dioxo-1H-1,4-benzodiazepine}-4-propanoic acid;
3-{4H-imidazo[1,2-a][1,4]benzodiazepine-5(6H)-1-methyl-6-oxo-9-[[[(benzimidazol-2-yl)methyl]methylamino]carbonyl]}-4-propanoic acid;
4-[4-[2-(1H-benzimidazol-2-yl)ethyl]-1-piperazinyl]-1-piperidineacetic acid;
1-hydroxy-4-[4-[3-(1H-benzimidazol-2-yl)propyl]-1-piperazinyl]-cyclohexaneacetic acid;
N-[3-[1-[[2-(2-Benzimidazolyl)ethyl]carbonyl]piperidinyl]carbonyl]-.beta.-alanine;
2-[(Benzimidazol-2-yl)methyl]-5-[2-(carboxy-ethyl)amino]carbonyl]
-2,3-dihydro-3-oxo-1H-isoindole;
[3(R)-[2-(benzimidazol-2-yl)ethyl]-2-oxopiperidinyl]acetyl-3(R)-methyl-.beta.-alanine;
4-[[[[2-(benzimidazolyl)methyl]carbonyl]methylamino]-acetyl]phenoxyacetic acid;
4-[[[[2-(Benzimidazolyl)methyl]carbonyl]methylamino]acetyl]-1,2-phenylenedioxydiacetic acid;
N-[3-[[[(2-Benzimidazolyl)methyl]carbonyl]amino]benzoyl]-.beta.-alanine;
[[1-[N-[[(2-Benzimidazolyl)methyl]carbonyl]tyrosyl]-4-piperidinyl]oxy]acetic acid;
(S)-4-[[[(2-Benzimidazolyl)methyl]carbonyl]glycyl]-3-methoxycarbonylmethyl-2-oxopiperazine-1-acetic acid;
(3S,5S)-5-[[4-[(2-Benzimidazolyl)methyl]phenyl]oxymethyl]-3-carboxymethyl-2-pyrrolidinone;
1-[(2-Benzimidazolyl)methyl]-3-[4-(2-carboxyethyl)phenyl]-4-methoxy-3-pyrrolin-2-one;
2-[6-(benzimidazol-2-yl) methylaminocarbonyl)-1,2,3,4-tetrahydroisoquinolinyl]acetic acid;
2-[6-(benzimidazol-2-yl)methylaminocarbonyl)-1-oxo-1,2,3,4-tetrahydroisoquinolinyl]acetic acid;
2-[6-((benzimidazol-2-yl)methylcarbonylamino)tetralin]acetic acid;
2-[6-((benzimidazol-2-yl)methylaminocarbonyl)tetralin]acetic acid;
2-[5-((benzimidazol-2-yl)methylcarbonylamino)-benzofuran]-propionic acid;
2-[5-((benzimidazol-2-yl)methylcarbonylamino)-2,3-dihydro-benzofuran]-propionic acid;
2-[5-(6-aminopyridinyl-2-methylaminocarbonyl)-benzofuran]-propionic acid;
2-[5-((benzimidazol-2-yl)methylaminocarbonyl)-2,3-dihydro-benzofuran]-propionic acid; or 1-[(1H-benzimidazol-2-yl)methyl]-3-{4[(2-ethoxycarbonyl)ethyl)]phenyl}-3-oxo-imidazolidine;
or pharmaceutically acceptable salts thereof.
3-[[3-[2-(benzimidazol-2-yl)ethyl]isoxazolin-5(R,S)-yl]acetyl]amino-3(R,S)-melhylpropanoic acid;
3-{3,4-dihydro-8-[[[(benzimidazol-2-yl)methyl]methylamino]carbonyl]-1-methyl-2,5-dioxo-1H-1,4-benzodiazepine}-4-propanoic acid;
3-{4H-imidazo[1,2-a][1,4]benzodiazepine-5(6H)-1-methyl-6-oxo-9-[[[(benzimidazol-2-yl)methyl]methylamino]carbonyl]}-4-propanoic acid;
4-[4-[2-(1H-benzimidazol-2-yl)ethyl]-1-piperazinyl]-1-piperidineacetic acid;
1-hydroxy-4-[4-[3-(1H-benzimidazol-2-yl)propyl]-1-piperazinyl]-cyclohexaneacetic acid;
N-[3-[1-[[2-(2-Benzimidazolyl)ethyl]carbonyl]piperidinyl]carbonyl]-.beta.-alanine;
2-[(Benzimidazol-2-yl)methyl]-5-[2-(carboxy-ethyl)amino]carbonyl]
-2,3-dihydro-3-oxo-1H-isoindole;
[3(R)-[2-(benzimidazol-2-yl)ethyl]-2-oxopiperidinyl]acetyl-3(R)-methyl-.beta.-alanine;
4-[[[[2-(benzimidazolyl)methyl]carbonyl]methylamino]-acetyl]phenoxyacetic acid;
4-[[[[2-(Benzimidazolyl)methyl]carbonyl]methylamino]acetyl]-1,2-phenylenedioxydiacetic acid;
N-[3-[[[(2-Benzimidazolyl)methyl]carbonyl]amino]benzoyl]-.beta.-alanine;
[[1-[N-[[(2-Benzimidazolyl)methyl]carbonyl]tyrosyl]-4-piperidinyl]oxy]acetic acid;
(S)-4-[[[(2-Benzimidazolyl)methyl]carbonyl]glycyl]-3-methoxycarbonylmethyl-2-oxopiperazine-1-acetic acid;
(3S,5S)-5-[[4-[(2-Benzimidazolyl)methyl]phenyl]oxymethyl]-3-carboxymethyl-2-pyrrolidinone;
1-[(2-Benzimidazolyl)methyl]-3-[4-(2-carboxyethyl)phenyl]-4-methoxy-3-pyrrolin-2-one;
2-[6-(benzimidazol-2-yl) methylaminocarbonyl)-1,2,3,4-tetrahydroisoquinolinyl]acetic acid;
2-[6-(benzimidazol-2-yl)methylaminocarbonyl)-1-oxo-1,2,3,4-tetrahydroisoquinolinyl]acetic acid;
2-[6-((benzimidazol-2-yl)methylcarbonylamino)tetralin]acetic acid;
2-[6-((benzimidazol-2-yl)methylaminocarbonyl)tetralin]acetic acid;
2-[5-((benzimidazol-2-yl)methylcarbonylamino)-benzofuran]-propionic acid;
2-[5-((benzimidazol-2-yl)methylcarbonylamino)-2,3-dihydro-benzofuran]-propionic acid;
2-[5-(6-aminopyridinyl-2-methylaminocarbonyl)-benzofuran]-propionic acid;
2-[5-((benzimidazol-2-yl)methylaminocarbonyl)-2,3-dihydro-benzofuran]-propionic acid; or 1-[(1H-benzimidazol-2-yl)methyl]-3-{4[(2-ethoxycarbonyl)ethyl)]phenyl}-3-oxo-imidazolidine;
or pharmaceutically acceptable salts thereof.
27. A pharmaceutical composition which comprises a pharmaceutically acceptable carrier and a compound according to any one of claims 1-26.
28. A method of inhibiting a vitronectin receptors in a mammal which comprises administering an effective amount of a compound according to formula (I) or (II) or (III) or (IV) or (V) as defined in claim 1.
29. A method according to claim 28 wherein the compound inhibits the vitronectinreceptor with a Ki at the vitronectin receptor that is ten-fold greater than the Ki for said compound at the fibrinogen receptor.
30. A method according to claim 28 wherein the compound inhibits the vitronectinreceptor with a Ki at the vitronectin receptor that is thirty-fold greater than the Ki for said compound at the fibrinogen receptor.
31. A method according to claim 28 wherein the compound inhibits the vitronectinreceptor with a Ki at the vitronectin receptor that is a hundred-fold greater than the Ki for said compound at the fibrinogen receptor.
32. A method according to claim 28 for treating diseases wherein bone resorption is a factor.
33. A method according to claim 28 for treating osteoporosis, inflammation, restenosis or atherosclerosis.
34. A method of inhibiting a vitronectin receptor in a mammal which comprises administering an effective amount of a compound according to formula (XXI) or (XXII) as defined in claim 13.
35. A method according to claim 34 wherein the compound inhibits the vitronectinreceptor with a Ki at the vitronectin receptor that is ten-fold greater than the Ki for said compound at the fibrinogen receptor.
36. A method according to claim 34 wherein the compound inhibits the vitronectinreceptor with a Ki at the vitronectin receptor that is thirty-fold greater than the Ki for said compound at the fibrinogen receptor.
37. A method according to claim 34 wherein the compound inhibits the vitronectinreceptor with a Ki at the vitronectin receptor that is a hundred-fold greater than the Ki for said compound at the fibrinogen receptor.
38. A method according to claim 34 for treating diseases wherein bone resorption is a factor.
39. A method according to claim 34 for treating osteoporosis, inflammation, restenosis or atherosclerosis.
40. A method of inhibiting a vitronectin receptor in a mammal which comprises administering an effective amount of a compound according to claim 25.
41. A method according to claim 40 wherein the compound inhibits the vitronectinreceptor with a Ki at the vitronectin receptor that is ten-fold greater than the Ki for said compound at the fibrinogen receptor.
42. A method according to claim 40 wherein the compound inhibits the vitronectinreceptor with a Ki at the vitronectin receptor that is thirty-fold greater than the Ki for said compound at the fibrinogen receptor.
43. A method according to claim 40 wherein the compound inhibits the vitronectinreceptor with a Ki at the vitronectin receptor that is a hundred-fold greater than the Ki for said compound at the fibrinogen receptor.
44. A method according to claim 40 for treating diseases wherein bone resorption is a factor.
45. A method according to claim 40 for treating osteoporosis, inflammation, restenosis or atherosclerosis.
46. The use of a compound according to any one of claims 1-26 in the manufactureof a medicament.
47. The use of a compound according to any one of claims 1-26 in the manufactureof a medicament for the inhibition of the vitronectin receptor in a mammal in need thereof.
48. The use of a compound according to any one of claims 1-26 in the manufactureof a medicament for the treatment of diseases in which bone resorption is a factor.
49. The use of a compound according to any one of claims 1-26 in the manufactureof a medicament for the treatment of osteoporosis, inflammation, restenosis, or atherosclerosis.
50. A process for preparing a compound of the formula (I) or (II) or (III) or (IV) or (V):
or or or or wherein:
W is - (CHRg)b-V'- or phenyl;
A is a fibrinogen receptor antagonist template;
V' is CONR21 or NR21CO;
G is NRe,S or O;
Rg is H, C1-6alkyl, Het-C0-6alkyl, C3-7cycloalkyl-C0-6alkyl or Ar- C0-6alkyl;
R21 is Het-(CH2)0-6-U'-(CH2)1-6-, C3-7cycloalkyl-(CH2)0-6-U'-(CH2)1-6-, or Ar-(CH2)0-6-U'-(CH2)1-6-;
U' is CONRf or NRfCO;
Rf is H, C1-6alkyl or Ar-C1-6alkyl;
Re is H, C1-6alkyl, Ar-C1-6alkyl, Het-C1-6alkyl, C3-7cycloalkyl-C1-6alkyl, (CH2)qOH or (CH2)kCO2Rg;
k is 0, 1 or 2;
q is 1 or 2;
b is 0, 1 or 2;
Rb and Rc are independently selected from H, C1-6alkyl, Ar-C0-6alkyl, Het-C0-6alkyl, or C3-6cycloalkyl-C0-6alkyl, halogen, CF3, ORf, S(O)kRf, CORf, NO2, N(Rf)2, CO(NRf)2, CH2N(Rf)2, or Rb and Rc are joined together to form a five or six membered aromatic or non-aromatic carbocyclic or heterocyclic ring, optionally substituted by up to three substituents chosen from halogen, CF3, C1-4alkyl, ORf, S(O)kRf, CORf, CO2Rf OH, NO2, N(Rf)2, CO(NRf)2, and CH2N(Rf)2, or methylenedioxy;
or a pharmecutically acceptable salt thereof, which process comprises:
(i) for formula (I) compounds, reacting a compound of the formula (Ia) with a compound of formula (Ib):
wherein:
Rb, Rc, Rf, and A are as defined in formula (I), with any reactive functional groups protected; and L' and L" are groups which react to form an amide bond in the moiety W or L' is phenyl substituted by -SnBu3, and L" is halo;
and thereafter removing any protecting groups, and optionally forming a pharmaceutically acceptable salt; or (ii) for formula (II) compounds, reacting a compound of the formula (IIa) with a compound of the formula (Ib):
L"-A
(Ib) wherein:
Rb, Rc, G and A are as defined in formula (I), with any reactive functional groups protected; and L' and L" are groups which react to form an amide bond in the moiety W or L' is phenyl substituted by -SnBu3, and L" is halo;
and thereafter removing any protecting groups, and optionally forming a pharmaceutically acceptable salt; or (iii) for formula (III) compounds, reacting a compound of the formula (IIIa) with a compound of the formula (Ib):
L"-A
(Ib) wherein:
Rb, Rc, Re, and A are as defined in formula(I), with any reactive functional groups protected; and L' and L" are groups which react to form an amide bond in the moiety W or L' is phenyl substituted by -SnBu3, and L" is halo;
and thereafter removing any protecting groups, and optionally forming a pharmaceutically acceptable salt; or (iv) for formula (IV) compounds, reacting a compound of the formula (IVa) with a compound of the formula (Ib):
L"-A
(Ib) wherein:
Rg, Re, and A are as defined in formula(I), with any reactive functional groups protected; and L' and L" are groups which react to form an amide bond in the moiety W or L' is phenyl substituted by -SnBu3, and L" is halo;
and thereafter removing any protecting groups, and optionally forming a pharmaceutically acceptable salt; or (v) for formula (V) compounds, reacting a compound of the formula (Va) with a compound of the formula (Ib):
L"-A
(Ib) wherein:
Rg, Re, and A are as defined in formula(I), with any reactive functional groups protected; and L' and L" are groups which react to form an amide bond in the moiety W or L' is phenyl substituted by -SnBu3, and L" is halo;
and thereafter removing any protecting groups, and optionally forming a pharmaceutically acceptable salt.
or or or or wherein:
W is - (CHRg)b-V'- or phenyl;
A is a fibrinogen receptor antagonist template;
V' is CONR21 or NR21CO;
G is NRe,S or O;
Rg is H, C1-6alkyl, Het-C0-6alkyl, C3-7cycloalkyl-C0-6alkyl or Ar- C0-6alkyl;
R21 is Het-(CH2)0-6-U'-(CH2)1-6-, C3-7cycloalkyl-(CH2)0-6-U'-(CH2)1-6-, or Ar-(CH2)0-6-U'-(CH2)1-6-;
U' is CONRf or NRfCO;
Rf is H, C1-6alkyl or Ar-C1-6alkyl;
Re is H, C1-6alkyl, Ar-C1-6alkyl, Het-C1-6alkyl, C3-7cycloalkyl-C1-6alkyl, (CH2)qOH or (CH2)kCO2Rg;
k is 0, 1 or 2;
q is 1 or 2;
b is 0, 1 or 2;
Rb and Rc are independently selected from H, C1-6alkyl, Ar-C0-6alkyl, Het-C0-6alkyl, or C3-6cycloalkyl-C0-6alkyl, halogen, CF3, ORf, S(O)kRf, CORf, NO2, N(Rf)2, CO(NRf)2, CH2N(Rf)2, or Rb and Rc are joined together to form a five or six membered aromatic or non-aromatic carbocyclic or heterocyclic ring, optionally substituted by up to three substituents chosen from halogen, CF3, C1-4alkyl, ORf, S(O)kRf, CORf, CO2Rf OH, NO2, N(Rf)2, CO(NRf)2, and CH2N(Rf)2, or methylenedioxy;
or a pharmecutically acceptable salt thereof, which process comprises:
(i) for formula (I) compounds, reacting a compound of the formula (Ia) with a compound of formula (Ib):
wherein:
Rb, Rc, Rf, and A are as defined in formula (I), with any reactive functional groups protected; and L' and L" are groups which react to form an amide bond in the moiety W or L' is phenyl substituted by -SnBu3, and L" is halo;
and thereafter removing any protecting groups, and optionally forming a pharmaceutically acceptable salt; or (ii) for formula (II) compounds, reacting a compound of the formula (IIa) with a compound of the formula (Ib):
L"-A
(Ib) wherein:
Rb, Rc, G and A are as defined in formula (I), with any reactive functional groups protected; and L' and L" are groups which react to form an amide bond in the moiety W or L' is phenyl substituted by -SnBu3, and L" is halo;
and thereafter removing any protecting groups, and optionally forming a pharmaceutically acceptable salt; or (iii) for formula (III) compounds, reacting a compound of the formula (IIIa) with a compound of the formula (Ib):
L"-A
(Ib) wherein:
Rb, Rc, Re, and A are as defined in formula(I), with any reactive functional groups protected; and L' and L" are groups which react to form an amide bond in the moiety W or L' is phenyl substituted by -SnBu3, and L" is halo;
and thereafter removing any protecting groups, and optionally forming a pharmaceutically acceptable salt; or (iv) for formula (IV) compounds, reacting a compound of the formula (IVa) with a compound of the formula (Ib):
L"-A
(Ib) wherein:
Rg, Re, and A are as defined in formula(I), with any reactive functional groups protected; and L' and L" are groups which react to form an amide bond in the moiety W or L' is phenyl substituted by -SnBu3, and L" is halo;
and thereafter removing any protecting groups, and optionally forming a pharmaceutically acceptable salt; or (v) for formula (V) compounds, reacting a compound of the formula (Va) with a compound of the formula (Ib):
L"-A
(Ib) wherein:
Rg, Re, and A are as defined in formula(I), with any reactive functional groups protected; and L' and L" are groups which react to form an amide bond in the moiety W or L' is phenyl substituted by -SnBu3, and L" is halo;
and thereafter removing any protecting groups, and optionally forming a pharmaceutically acceptable salt.
51. A process for preparing a compound of the formula (XXI):
wherein A and B are as defined in claim 13, which process comprises reacting a compound of the formula (XXV) with a compound of the formula (XXVI):
(XXVI) wherein:
A is as defined in claim 13, with any reactive functional groups protected;
and L3 and L4 are groups which react to form a covalent bond in the moiety B;
and thereafter removing any protecting groups, and optionally forming a pharmaceutically acceptable salt.
wherein A and B are as defined in claim 13, which process comprises reacting a compound of the formula (XXV) with a compound of the formula (XXVI):
(XXVI) wherein:
A is as defined in claim 13, with any reactive functional groups protected;
and L3 and L4 are groups which react to form a covalent bond in the moiety B;
and thereafter removing any protecting groups, and optionally forming a pharmaceutically acceptable salt.
52. A process for preparing a compound of the formula (XXI):
wherein A, B and G are as defined in claim 13, which process comprises reacting a compound of the formula (XXVII) with a compound of the formula (XXVIII):
(XXXVIII) wherein:
A and G are as defined herinabove, with any reactive functional groups protected; and L5 and L6 are groups which react to form a covalent bond in the moiety B;
and thereafter removing any protecting groups, and optionally forming a pharmaceutically acceptable salt.
wherein A, B and G are as defined in claim 13, which process comprises reacting a compound of the formula (XXVII) with a compound of the formula (XXVIII):
(XXXVIII) wherein:
A and G are as defined herinabove, with any reactive functional groups protected; and L5 and L6 are groups which react to form a covalent bond in the moiety B;
and thereafter removing any protecting groups, and optionally forming a pharmaceutically acceptable salt.
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| US936695P | 1995-12-29 | 1995-12-29 | |
| US60/009,366 | 1995-12-29 |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CA2241633A1 true CA2241633A1 (en) | 1997-07-10 |
Family
ID=21737205
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CA002241633A Abandoned CA2241633A1 (en) | 1995-12-29 | 1996-12-20 | Vitronectin receptor antagonists |
Country Status (16)
| Country | Link |
|---|---|
| EP (1) | EP0869787A4 (en) |
| JP (1) | JP2000502354A (en) |
| KR (1) | KR19990076878A (en) |
| CN (1) | CN1209744A (en) |
| AU (1) | AU1354097A (en) |
| BR (1) | BR9612327A (en) |
| CA (1) | CA2241633A1 (en) |
| CZ (1) | CZ203698A3 (en) |
| HU (1) | HUP9900754A3 (en) |
| IL (1) | IL125033A0 (en) |
| MX (1) | MX9805255A (en) |
| NO (1) | NO983003L (en) |
| PL (1) | PL327694A1 (en) |
| TR (1) | TR199801253T2 (en) |
| WO (1) | WO1997024119A1 (en) |
| ZA (1) | ZA9610859B (en) |
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| US20030125317A1 (en) | 1996-10-02 | 2003-07-03 | Smithkline Beecham Corporation | Vitronectin receptor antagonists |
| DE19653645A1 (en) * | 1996-12-20 | 1998-06-25 | Hoechst Ag | Vitronectin receptor antagonists, their preparation and their use |
| US6482821B2 (en) | 1996-12-20 | 2002-11-19 | Hoechst Aktiengellschaft | Vitronectin receptor antagonists, their preparation and their use |
| US6218387B1 (en) * | 1996-12-20 | 2001-04-17 | Hoechst Aktiengesellschaft | Vitronectin receptor anatagonists, their preparation and their use |
| DE19653647A1 (en) * | 1996-12-20 | 1998-06-25 | Hoechst Ag | Vitronectin receptor antagonists, their preparation and their use |
| WO1999006049A1 (en) * | 1997-08-04 | 1999-02-11 | Smithkline Beecham Corporation | Integrin receptor antagonists |
| WO1999030709A1 (en) * | 1997-12-17 | 1999-06-24 | Merck & Co., Inc. | Integrin receptor antagonists |
| CN1140511C (en) | 1998-04-09 | 2004-03-03 | 明治制果株式会社 | Amino piperidine derivatives as integrin alpha beta 3 antagonists |
| ES2229718T3 (en) | 1998-06-12 | 2005-04-16 | Societe De Conseils De Recherches Et D'applications Scientifiques S.A.S. | DERIVATIVES OF IMIDAZOLIL AND ITS USE AS LIGANDS OF SOMATOSTATIN RECEPTORS. |
| SE9803518D0 (en) * | 1998-10-15 | 1998-10-15 | Astra Pharma Prod | Novel compounds |
| US6339083B1 (en) * | 1998-12-14 | 2002-01-15 | Bayer Aktiengesellschaft | Multiheterocyclic pharmAceuticals |
| CA2358855A1 (en) | 1999-02-03 | 2000-08-10 | Merck & Co., Inc. | Benzazepine derivatives as alpha-v integrin receptor antagonists |
| US6380210B1 (en) | 1999-04-02 | 2002-04-30 | Neurogen Corporation | Heteroaryl fused aminoalkyl-imidazole derivatives: selective modulators of GABAa receptors |
| US6358949B1 (en) | 1999-04-02 | 2002-03-19 | Neurogen Corporation | Aryl and hetroaryl fused aminoalkyl-imidazole derivatives: selective modulators of bradykinin B2 receptors |
| US6627624B1 (en) | 1999-04-02 | 2003-09-30 | Neurogen Corporation | Aryl fused aminoalkyl-imidazole derivatives: selective modulators of GABAa receptors |
| MXPA01009949A (en) * | 1999-04-02 | 2003-07-14 | Neurogen Corp | ARYL AND HETEROARYL FUSED AMINOALKYL-IMIDAZOLE DERIVATIVES: SELECTIVE MODULATORS OF GABAa RECEPTORS. |
| US6271241B1 (en) | 1999-04-02 | 2001-08-07 | Neurogen Corporation | Cycloalkyl and aryl fused aminoalkyl-imidazole derivatives: modulators and GLP-1 receptors |
| US6281237B1 (en) | 1999-04-02 | 2001-08-28 | Neurogen Corporation | N-phenyl benzimidazolecarboxamide and N-phenyl indolecarboxamide derivatives |
| GB9908355D0 (en) * | 1999-04-12 | 1999-06-09 | Rhone Poulenc Rorer Ltd | Chemical compounds |
| WO2001004103A1 (en) | 1999-07-13 | 2001-01-18 | F. Hoffmann-La Roche Ag | Benzazepinones and quinazolines |
| CN1374863A (en) | 1999-07-21 | 2002-10-16 | 惠氏公司 | Bicyclic antagonists selective for the alpha beta integrin |
| US6750219B1 (en) | 1999-08-05 | 2004-06-15 | Meiji Seika Kaisha, Ltd. | Ω-amino-α-hydroxycarboxylic acid derivatives having integrin ανβ3 antagonistic activity |
| JP2003506452A (en) * | 1999-08-06 | 2003-02-18 | スミスクライン・ビーチャム・コーポレイション | Vitronectin receptor antagonists useful for the treatment of stroke |
| WO2001056995A1 (en) | 2000-01-18 | 2001-08-09 | Nuerogen Corporation | Substituted imidazoles as selective modulators of bradykinin b2 receptors |
| US6545029B2 (en) | 2000-06-12 | 2003-04-08 | Bayer Aktiengesellschaft | Phenylserine derivatives as integrin antagonists |
| DE60128102T2 (en) | 2000-08-01 | 2008-01-03 | Societe De Conseils De Recherches Et D`Applications Scientifiques (S.C.R.A.S.) | IMIDAZOLE DERIVATIVES |
| RU2316337C2 (en) | 2001-04-24 | 2008-02-10 | Мерк Патент Гмбх | Combined therapy by using antiangiogenic agents and tnf-alpha |
| US7030150B2 (en) * | 2001-05-11 | 2006-04-18 | Trimeris, Inc. | Benzimidazole compounds and antiviral uses thereof |
| EP2428226A1 (en) | 2001-10-22 | 2012-03-14 | The Scripps Research Institute | Antibody targeting compounds |
| MY150088A (en) | 2003-05-19 | 2013-11-29 | Irm Llc | Immunosuppressant compounds and compositions |
| MXPA05012460A (en) * | 2003-05-19 | 2006-05-25 | Irm Llc | Immunosuppressant compounds and compositions. |
| EP1732905A1 (en) | 2004-03-31 | 2006-12-20 | Lexicon Genetics Incorporated | 2-aminomethylthiazole-5-carboxamides as protein kinase modulators |
| UA87854C2 (en) | 2004-06-07 | 2009-08-25 | Мерк Энд Ко., Инк. | N-(2-benzyl)-2-phenylbutanamides as androgen receptor modulators |
| ES2401138T3 (en) | 2004-08-13 | 2013-04-17 | Genentech, Inc. | Thiazole-based enzyme inhibitors that use ATP |
| WO2007084670A2 (en) | 2006-01-18 | 2007-07-26 | Merck Patent Gmbh | Specific therapy using integrin ligands for treating cancer |
| MX2009007597A (en) | 2007-01-18 | 2009-07-22 | Merck Patent Gmbh | Specific therapy and medicament using integrin ligands for treating cancer. |
| WO2008141081A1 (en) | 2007-05-10 | 2008-11-20 | Amr Technology, Inc. | Aryl- and heteroaryl-substituted tetrahydrobenzo-1,4-diazepines and use thereof to block reuptake of norepinephrine, dopamine, and serotonin |
| EP2395992A2 (en) | 2009-02-10 | 2011-12-21 | The Scripps Research Institute | Chemically programmed vaccination |
| EA201101651A1 (en) | 2009-05-25 | 2012-08-30 | Мерк Патент Гмбх | CONTINUOUS INTRODUCTION OF INTEGRINE LIGANDS FOR CANCER TREATMENT |
| CA2950390C (en) | 2014-05-30 | 2020-09-22 | Pfizer Inc. | Carbonitrile derivatives as selective androgen receptor modulators |
| EP3684767B1 (en) | 2017-09-22 | 2024-04-24 | Jubilant Epipad LLC | Heterocyclic compounds as pad inhibitors |
| FI3697785T3 (en) | 2017-10-18 | 2023-04-03 | Jubilant Epipad LLC | Imidazo-pyridine compounds as pad inhibitors |
| CN111386265A (en) | 2017-11-06 | 2020-07-07 | 朱比连特普罗德尔有限责任公司 | Pyrimidine derivatives as inhibitors of PD1/PD-L1 activation |
| IL274762B2 (en) | 2017-11-24 | 2023-10-01 | Jubilant Episcribe Llc | Novel heterocyclic compounds as prmt5 inhibitors |
| KR20200131845A (en) | 2018-03-13 | 2020-11-24 | 주빌런트 프로델 엘엘씨 | Bicyclic compounds as inhibitors of PD1/PD-L1 interaction/activation |
| WO2023275715A1 (en) | 2021-06-30 | 2023-01-05 | Pfizer Inc. | Metabolites of selective androgen receptor modulators |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| US3627754A (en) * | 1970-05-13 | 1971-12-14 | Hoffmann La Roche | Process for preparing 7-lower alkanoyl benzodiazepines utilizing ceric salts |
| US5250679A (en) * | 1991-10-18 | 1993-10-05 | Genentech, Inc. | Nonpeptidyl platelet aggregation inhibitors having specificity for the GPIIb III.sub. receptor |
-
1996
- 1996-12-20 KR KR1019980705005A patent/KR19990076878A/en not_active Application Discontinuation
- 1996-12-20 JP JP9524557A patent/JP2000502354A/en active Pending
- 1996-12-20 TR TR1998/01253T patent/TR199801253T2/en unknown
- 1996-12-20 PL PL96327694A patent/PL327694A1/en unknown
- 1996-12-20 WO PCT/US1996/020748 patent/WO1997024119A1/en not_active Application Discontinuation
- 1996-12-20 IL IL12503396A patent/IL125033A0/en unknown
- 1996-12-20 EP EP96945087A patent/EP0869787A4/en not_active Withdrawn
- 1996-12-20 HU HU9900754A patent/HUP9900754A3/en unknown
- 1996-12-20 CN CN96180113A patent/CN1209744A/en active Pending
- 1996-12-20 BR BR9612327A patent/BR9612327A/en unknown
- 1996-12-20 CA CA002241633A patent/CA2241633A1/en not_active Abandoned
- 1996-12-20 CZ CZ982036A patent/CZ203698A3/en unknown
- 1996-12-20 AU AU13540/97A patent/AU1354097A/en not_active Abandoned
- 1996-12-23 ZA ZA9610859A patent/ZA9610859B/en unknown
-
1998
- 1998-06-26 MX MX9805255A patent/MX9805255A/en unknown
- 1998-06-26 NO NO983003A patent/NO983003L/en not_active Application Discontinuation
Also Published As
| Publication number | Publication date |
|---|---|
| AU1354097A (en) | 1997-07-28 |
| KR19990076878A (en) | 1999-10-25 |
| JP2000502354A (en) | 2000-02-29 |
| CN1209744A (en) | 1999-03-03 |
| ZA9610859B (en) | 1997-10-24 |
| TR199801253T2 (en) | 1998-12-21 |
| NO983003D0 (en) | 1998-06-26 |
| CZ203698A3 (en) | 1999-05-12 |
| NO983003L (en) | 1998-08-26 |
| IL125033A0 (en) | 1999-01-26 |
| MX9805255A (en) | 1998-11-29 |
| BR9612327A (en) | 1999-07-13 |
| EP0869787A4 (en) | 1999-03-24 |
| EP0869787A1 (en) | 1998-10-14 |
| WO1997024119A1 (en) | 1997-07-10 |
| HUP9900754A2 (en) | 1999-07-28 |
| PL327694A1 (en) | 1998-12-21 |
| HUP9900754A3 (en) | 1999-11-29 |
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