AU721130B2 - Fibrinogen receptor antagonists - Google Patents

Fibrinogen receptor antagonists Download PDF

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AU721130B2
AU721130B2 AU35798/97A AU3579897A AU721130B2 AU 721130 B2 AU721130 B2 AU 721130B2 AU 35798/97 A AU35798/97 A AU 35798/97A AU 3579897 A AU3579897 A AU 3579897A AU 721130 B2 AU721130 B2 AU 721130B2
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AU3579897A (en
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Mark E. Duggan
Melissa S. Egbertson
George D. Hartman
Nathan C. Ihle
Steven D. Young
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Merck and Co Inc
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Merck and Co Inc
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    • C07D213/04—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members having three double bonds between ring members or between ring members and non-ring members having no bond between the ring nitrogen atom and a non-ring member or having only hydrogen or carbon atoms directly attached to the ring nitrogen atom
    • C07D213/60—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members having three double bonds between ring members or between ring members and non-ring members having no bond between the ring nitrogen atom and a non-ring member or having only hydrogen or carbon atoms directly attached to the ring nitrogen atom with hetero atoms or with carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals, directly attached to ring carbon atoms
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    • C07D211/18—Heterocyclic compounds containing hydrogenated pyridine rings, not condensed with other rings with only hydrogen or carbon atoms directly attached to the ring nitrogen atom having no double bonds between ring members or between ring members and non-ring members with hydrocarbon or substituted hydrocarbon radicals directly attached to ring carbon atoms with substituted hydrocarbon radicals attached to ring carbon atoms
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    • C07D213/04—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members having three double bonds between ring members or between ring members and non-ring members having no bond between the ring nitrogen atom and a non-ring member or having only hydrogen or carbon atoms directly attached to the ring nitrogen atom
    • C07D213/24—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members having three double bonds between ring members or between ring members and non-ring members having no bond between the ring nitrogen atom and a non-ring member or having only hydrogen or carbon atoms directly attached to the ring nitrogen atom with substituted hydrocarbon radicals attached to ring carbon atoms
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    • C07D213/02—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members having three double bonds between ring members or between ring members and non-ring members
    • C07D213/04—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members having three double bonds between ring members or between ring members and non-ring members having no bond between the ring nitrogen atom and a non-ring member or having only hydrogen or carbon atoms directly attached to the ring nitrogen atom
    • C07D213/60—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members having three double bonds between ring members or between ring members and non-ring members having no bond between the ring nitrogen atom and a non-ring member or having only hydrogen or carbon atoms directly attached to the ring nitrogen atom with hetero atoms or with carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals, directly attached to ring carbon atoms
    • C07D213/72—Nitrogen atoms
    • C07D213/73—Unsubstituted amino or imino radicals
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    • C07D213/04—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members having three double bonds between ring members or between ring members and non-ring members having no bond between the ring nitrogen atom and a non-ring member or having only hydrogen or carbon atoms directly attached to the ring nitrogen atom
    • C07D213/60—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members having three double bonds between ring members or between ring members and non-ring members having no bond between the ring nitrogen atom and a non-ring member or having only hydrogen or carbon atoms directly attached to the ring nitrogen atom with hetero atoms or with carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals, directly attached to ring carbon atoms
    • C07D213/78—Carbon atoms having three bonds to hetero atoms, with at the most one bond to halogen, e.g. ester or nitrile radicals
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    • C07D295/04—Heterocyclic compounds containing polymethylene-imine rings with at least five ring members, 3-azabicyclo [3.2.2] nonane, piperazine, morpholine or thiomorpholine rings, having only hydrogen atoms directly attached to the ring carbon atoms with substituted hydrocarbon radicals attached to ring nitrogen atoms
    • C07D295/14—Heterocyclic compounds containing polymethylene-imine rings with at least five ring members, 3-azabicyclo [3.2.2] nonane, piperazine, morpholine or thiomorpholine rings, having only hydrogen atoms directly attached to the ring carbon atoms with substituted hydrocarbon radicals attached to ring nitrogen atoms substituted by carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals
    • C07D295/155—Heterocyclic compounds containing polymethylene-imine rings with at least five ring members, 3-azabicyclo [3.2.2] nonane, piperazine, morpholine or thiomorpholine rings, having only hydrogen atoms directly attached to the ring carbon atoms with substituted hydrocarbon radicals attached to ring nitrogen atoms substituted by carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals with the ring nitrogen atoms and the carbon atoms with three bonds to hetero atoms separated by carbocyclic rings or by carbon chains interrupted by carbocyclic rings
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    • C07D333/50—Heterocyclic compounds containing five-membered rings having one sulfur atom as the only ring hetero atom condensed with carbocyclic rings or ring systems
    • C07D333/52—Benzo[b]thiophenes; Hydrogenated benzo[b]thiophenes
    • C07D333/62—Benzo[b]thiophenes; Hydrogenated benzo[b]thiophenes with hetero atoms or with carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals, directly attached to carbon atoms of the hetero ring
    • C07D333/68—Carbon atoms having three bonds to hetero atoms with at the most one bond to halogen
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    • C07D401/04—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing two hetero rings directly linked by a ring-member-to-ring-member bond
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    • C07D471/02—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, at least one ring being a six-membered ring with one nitrogen atom, not provided for by groups C07D451/00 - C07D463/00 in which the condensed system contains two hetero rings
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Description

WO 98/00134 PCT/US97/11133 -1- TITLE OF THE INVENTION FIBRINOGEN RECEPTOR ANTAGONISTS BACKGROUND OF THE INVENTION The invention relates generally to modulating cell adhesion and to inhibiting the binding of fibrinogen and other proteins to blood platelets, and inhibiting the aggregation of blood platelets specifically to the gp IIb/Illa fibrinogen receptor site. Fibrinogen is a glycoprotein present in blood plasma that participates in platelet aggregation and in fibrin formation. Platelets are cell-like anucleated fragments, found in the blood of all mammals, that also participate in blood coagulation.
Interaction of fibrinogen with the IIb/IIIa receptor site is known to be essential for normal platelet function.
When a blood vessel is damaged by an injury or other causative factor, platelets adhere to the disrupted subendothethial surface. The adherent platelets subsequently release biologically active constituents and aggregate. Aggregation is initiated by the binding of agonists, such as thrombin, epinephrine, or ADP to specific platelet membrane receptors. Stimulation by agonists results in exposure of latent fibrinogen receptors on the platelet surface, and binding of fibrinogen to the glycoprotein IIb/IIIa receptor complex.
Attempts have been made to use natural products and synthetic peptides to determine the mechanism of adhesion and platelet aggregation. For example, Rouslahti and Pierschbacher in Science, 238, 491-497 (1987), describe adhesive proteins such as fibronectin, vitronectin, osteopontin, collagens, thrombospondin, fibrinogen, and von Willebrand factor that are present in extracellular matrices and in blood. The proteins contain the tripeptide arginine-glycine-aspartic acid (RGD) as their glycoprotein IIb/IIa recognition site. These arginineglycine-aspartic acid containing tripeptides are recognized by at least one member of a family of structurally related receptors, integrins, which are heterodimeric proteins with two membrane-spanning subunits. The authors state that the conformation of the tripeptide WO 98/00134 PCT/US97/11133 -2sequence in the individual proteins may be critical to recognition specificity.
Cheresh-in Proc. Nat'l Acad. Sci. 84, 6471-6475, (1987), describes an Arg-Gly-Asp directed adhesion receptor expressed by human endothethial cells that is structurally similar to the b/IlIa complex on platelets but is antigenically and functionally distinct. This receptor is directly involved in endothelial cell attachment to fibrinogen, von Willebrand factor, and vitronectin.
Pierschbacher and Rouslahti, in J. of Biol. Chem., 262, 17294-17298 (1987) hypothesized that the Arg-Gly-Asp sequence alone would be a sufficient signal for receptor recognition and binding and that, therefore, the conformation of the tri-peptide sequence would be determinative. Various synthetic peptides were produced and the authors concluded that the stereochemical conformation of Arg-Gly-Asp as influenced by enantiomeric substitutions or additions to this sequence significantly influenced receptor-ligand interaction. The authors further showed that cyclization of a decapeptide by forming a disulfide bridge between non-terminal residues Pen and Cys, rendered the peptide much less effective at inhibiting attachment to fibronectin.
In Proc. Nat'l Acad. Sci. 81, 5985-5988 (1984), the same authors describe tetrapeptide variants of the cell recognition site of fibronectin that retain attachment-promoting activity. Peptides having a tetrapeptide recognition site are described in U.S. Pat. Nos. 4,589,881 and 4,614,517. A number of large polypeptide fragments in the cellbinding domain of fibronectin have cell-attachment activity. For example, see U.S. Pat. Nos. 4,517,686, 4,661,111 and U.S. Pat. No.
4,578,079.
Ruggeri et al., Proc. Nat'l Acad. Sci. 83, 5708- 5712 (1986) explore a series of synthetic peptides designed in lengths to 16 residues, that contain RGD and a valine attached to the aspartic acid residue of RGD that inhibit fibrinogen binding to platelets. See also Koczewiak et al., Biochem. 23, 1767-1774 (1984); Ginsberg et al., WO 98/00134 PCT/US97/11133 -3- J. Biol. Chem. 260(7), 3931-3936 (1985); and Haverstick et al., Blood 66(4), 946-952 (1985). Other inhibitors are disclosed in Eur. Pat. App.
Nos. 275,748 and 298,820.
A number of low molecular weight polypeptide factors have been isolated from snake venom. These factors apparently have high affinity for the gp IIb/IIIa complex. For example, Huang et al., J.
Biol Chem., 262, 16157-16163 (1987); Huang et al., Biochemistry, 28, 661-666 (1989) describe the primary structure of the venom trigramin which is a 72 amino acid polypeptide that contains the RGD subunit.
Echistatin is another compound which has high affinity for the gp IIb/Ia complex. This polypeptide contains 49 amino acids and has the RGD subunit and various disulfide bridges. Gan et al., J. Biol. Chem., 263, 19827-19832 (1988). See also, Dennis et al., Proc. Nat'l Acad. Sci.
USA, 87, 2471-2475 (1989). However, these snake venom factors also have high affinity for other members of the adhesive protein receptor family including the vitronectin and fibronectin receptors so are not selective for the gp IIb/HIa complex.
While it is known that the tripeptide sequence Arg-Gly-Asp is present in certain polypeptides that can duplicate or inhibit the cell attachment-promoting effects of fibronectin and vitronectin, the tripeptide Arg-Gly-Asp has low activity. At present, there is little understanding of how other amino acids coupled to this sequence influence binding specificity. U.S. Pat. No 5,023,233 discloses small cyclic hexapeptides which contain the sequence Arg-Gly-Asp and are useful platelet aggregation inhibitors. U.S. Pat. No. 5,037,808 discloses the use of indolyl platelet-aggregation inhibitors which are believed to act by antagonizing interactions between fibrinogen and/or extracellular matrix proteins and the platelet gp IIb/Illa receptor. U.S. Pat. No.
5,037,808 discloses guanidino peptide mimetic compounds that retain an Asp residue which inhibit platelet aggregation. W09014103 describes the use of antibody-polypeptide conjugates wherein said polypeptides contain the Arg-Gly-Asp (RGD) sequence.
W09111458 discloses the use of large cyclic peptides containing RGD flanked by proline residues which are platelet aggregation inhibitors. WO 9101331 discloses small cyclic platelet aggregation inhibitors which are synthetic cyclic pentapeptides containing the tripeptide sequence Arg-Gly-Asp and a thioether linkage in the cycle. US Patent No. 5,051,405 also discloses the use of peptides and pseudopeptides such as N-amidino-piperidine-3-carboxylglycyl-L-aspartyl-L-valine that inhibit platelet aggregation and thrombus formation in mammalian blood. EP 445 796 discloses linear compounds which can include internal piperazinyl or piperidinyl derivatives.
EP 437 367 discloses linear polypeptide fibrinogen receptor antagonists. US Patent No. 5,256,812 discloses compounds of the formula R 1 -A-(W)a-X-(CH 2 B-Z-COOR wherein R 1 is a guandidino or amidino moiety and A and B are chosen from specific monosubstituted aryl or heterocyclic moieties.
While a multitude of compounds or peptide analogues believed to inhibit platelet aggregation by inhibiting binding to a blood platelet by fibrinogen are known, the present invention provides novel fibrinogen receptor antagonists that have significant binding activity and are, therefore, useful for the reasons stated herein. A number of very serious diseases and disorders involve hyperthrombotic complications which lead to intravascular thrombi and emboli. Myocardial infarction, stroke, phlebitis and a number of other series conditions create the need for novel and effective fibrinogen receptor antagonists.
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p.
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[R:\LIBAA]07996.doc:tab to 24 I0 PAGES 5 TO 24 HAVE BEEN LEFT INTENTIONALLY BLANK.
9 9 9 *9 4q 9b .9 99 9 p 99 9 9 9 9 9 9 9 *909 *999 9 9 9999 *999 .9 0 9 9*q* 9. 9 9 99 99 9 [R:\LIAA]07996.doc:tab Description of the Invention The invention includes compounds of the formula
X-Y-Z-A-B
and pharmaceutically acceptable salts, wherein X is HND HN NN
H
2 N N Y is
N
H
2
N
H
3 9* .9
SR
b 9e .9 9** *9 9 *ia *9 9 9 9 9
S
9O@S I, 9 i 9 0
S
S
S S is 5500 S. 0 5* S 00 0 [R:\LIBAA]07996.doc:tab WO 98/00134 WO 9800134PCTIUS97I1 1133 26 or X and Y form the fused ring systems:
HN
Z is 0 11
-CNH-
0 11
-C-
-NH-
-CH
2
CH
2
-CH=CH-
0
II
CH
3 0 11
-NHC-
Z represents a bond; WO 98/00134 WO 9800134PCTIUS97/11133 27 A is N HSO 2
O
6
H
5
OF
3
OH
3
OH
3 N HSO 2
CH
3
OCH
3 00H 3
CI
OH
3
CH
3
NO
2
OH
3 Br and B is -OCH2CO2H, -OCH2CO2CH2C(O)NHCH3, -OCH2CO2CH2C(O)N(CH2CH3)2, -OCH2CO2CH2CH3, -CH2CH2CO2H, WO 98/00134 WO 9800134PCT/US97/11 133 28 -CH2COOH, -OCH(CH-3)CO2H, or -OCH(CH3)CO2CH2CH3.
Exemplary compounds of the group include 3 -(4-(4-piperazin- 1-ylphenylcarbonylamino)phenyl) propanoic acid, 2-(4-(4-Piperazinyl- 1 -yl)phenylcarbonylamino)phenoxy) acetic acid, Ethyl 1-piperazinyl)phenylcarbonylamino)phenoxy)acetate, hydrochloride, -Piperazinyl)phenylcarbonylamino)indol- 1 -yl) acetic acid, 3 -(4-Piperazin- 1-ylphenyl)carbonylamnino)phenyl) propanoic acid, Ethyl 2-(4-(4-(piperazin- 1-yl)phenylcarbonytamino) phenoxy)propanoate 2-(4-(4-.(Piperazin- 1 -yl)phenylcarbonylamino)phenoxy) propionic acid, 2-(4-(((2-Piperazin- acid, 3 -Methyl-4-(( 1,2,3 ,4-tetrahydro-9H-pyrido[3 ,4-b]indol-7 yl)carbonylamino)phenoxyacetic acid, WO 98/00134 PCT/US97/11133 -29- -Dimethylethoxycarbonyl)-piperazin-1 -yl)-2thienylcarbonylamino)-3-methylphenoxy)-acetic acid, 4-(2-(1,1-dimethylethoxycarbonyl)-1,2,3,4-tetrahydro-9H-pyrido[3,4bjbenzofuran-7-yl)carbonylamino)-3-methylphenoxy acetic acid, 4-((2,3,4,5-tetrahydropyrazino-[1,2-a]indole-8-yl)carbonylamino)-3methylphenoxyacetic acid, and 4-((3-(1,1-Dimethylethoxycarbonyl)-l,la,2,3,4,5-hexahydropyrazino-[ 1,2-a]indole-8-yl)carbonylamino)-3-methylphenoxyacetic acid.
One test which is used to evaluate fibrinogen receptor antagonist activity is based on evaluation of inhibition of ADPstimulated platelets. Aggregation requires that fibrinogen bind to and occupy the platelet fibrinogen receptor site. Inhibitors of fibrinogen binding inhibit aggregation. In the ADP-stimulated platelet aggregation assay used to determine inhibition associated with the compounds claimed in the instant invention, human platelets are isolated from fresh blood, collected into acid citrate/dextrose by differential centrifugation followed by gel filtration on Sepharose 2B in divalent ion-free Tyrode's buffer (pH 7.4) containing 2% bovine serum albumin.
Platelet aggregation is measured at 37°C in a Chronolog aggregometer. The reaction mixture contains gel-filtered human platelets (2 x 108 per ml), fibrinogen (100 micrograms per ml (ug/ml)), Ca 2 (1 mM), and the compound to be tested. The aggregation is initiated by adding 10 mM ADP 1 minute after the other components are added. The reaction is then allowed to proceed for at least 2 minutes. The extent of inhibition of aggregation is expressed as the percentage of the rate of aggregation observed in the absence of inhibitor. The IC50 is the dose of a particular compound inhibiting aggregation by 50% relative to a control lacking the compound.
WO 98/00134 WO 9800134PCTIUS97/1 1133 The following compounds were tested and found to have values in the range between 10 nM and 50 mM: 3 -(4-(4-piperazin- 1 -ylphenylcarbonylamino)phenyl) propanoic acid, 2-(4-(4-Piperazinyl- 1 -yl)phenylcarbonylamino )phenoxy) acetic acid, Ethyl 1-piperazinyl)phenylcarbonylamrino)phenoxy)acetate, hydrochloride, 1 -Piperazinyl)phenylcarbonylamnino)indol- Il-yI) acetic acid, 3 -(4-Piperazin- 1-ylphenyl)carbonylamino)phenyl) propanoic acid, Ethyl 2-(4-(4-(piperazin- 1-yl)phenylcarbonylamino) phenoxy)propanoate 2-(4-(4-(Piperazin- I -yl)phenylcarbonylamino)phenoxy) propionic acid, 2-(4-(((2-Piperazin- I acid, 3 -Methyl-4-(( 1,2,3 ,4-tetrahydro-9H-pyrido [3 ,4-b]indol-7 yl)carbonylamino)phenoxyacetic acid, ,1 -Dimethylethoxycarbonyl)-piperazin- 1 -yl)-2thienylcarbonylamino)-3 -methylphenoxy)-acetic acid WO 98/00134 PCT/US97/11133 -31 -dimethylethoxycarbonyl)- 1,2,3,4-tetrahydro-9H-pyrido[3,4b]benzofuran-7-yl)carbonylamino)-3-methylphenoxy acetic acid, 4-((2,3,4,5-tetrahydropyrazino-[ 1,2-a]indole-8-yl)carbonylamino)-3methylphenoxyacetic acid, and 1 -Dimethylethoxycarbonyl)- 1,1 a,2,3,4,5-hexahydropyrazino-[ 1, 2 -a]indole-8-yl)carbonylamino)-3-methylphenoxyacetic acid.
Additionally, these compounds are useful for treating mammals suffering from a bone condition caused or mediated by increased bone resorption, who are in need of such therapy.
Pharmacologically effective amounts of the compounds, including pharamaceutically acceptable salts thereof, are administered to the mammal, to inhibit the activity of mammalian osteoclasts.
Additionally, these compounds are useful for treating angiogenesis (formation of new blood vessels). It has been postulated that the growth of tumors depends on an adequate blood supply, which in turn is dependent on the growth of new vessels into the tumor.
Inhibition of angiogenesis can cause tumor regression in animal models.
(See, Harrison's Principles of Internal Medicine, 12th ed., 1991). These compounds are therefore useful in the treatment of cancer by inhibiting tumor growth. (See Brooks et al., Cell, 79:1157-1164 (1994)).
The term "pharmaceutically acceptable salts" shall mean non-toxic salts of the compounds of this invention which are generally prepared by reacting the free base with a suitable organic or inorganic acid. Representative salts include the following salts: acetate, benzenesulfonate, benzoate, bicarbonate, bisulfate, bitartrate, borate, bromide, calcium edetate, camsylate, carbonate, chloride, clavulanate, citrate, dihydrochloride, edetate, edisylate, estolate, esylate, fumarate, gluceptate, gluconate, glutamate, glycollylarsanilate, hexylresorcinate, hydrabamine, hydrobromide, hydrochloride, hydroxynapthoate, iodide, isothionate, lactate, lactobionate, laurate, WO 98/00134 PCT/US97/11133 -32malate, maleate, mandelate, mesylate, methylbromide, methylnitrate, methylsulfate, mucate, napsylate, nitrate, oleate, oxalate, pamaote, palmitate, panthothenate, phosphate/diphosphate, polygalacturonate, salicylate, stearate, subacetate, succinate, tannate, tartrate, teoclate, tosylate, triethiodide, valerate.
Compounds of the present invention may be chiral; included within the scope of the present invention are racemic mixtures and separated enantiomers of the general formula. Furthermore, all diastereomers, including E, Z isomers, of the general formula are included in the present scope. Furthermore, hydrates as well as anhydrous compositions and polymorphs of the general formula are within the present invention.
Prodrugs, such as ester derivatives of described compounds, are compound derivatives which, when absorbed into the bloodstream of a warm-blooded animal, cleave in such a manner as to release the drug form and permit the drug to afford improved therapeutic efficacy.
The term "pharmaceutically effective amount" shall mean that amount of a drug or pharmaceutical agent that will elicit the biological or medical response of a tissue, system or animal that is being sought by a researcher or clinician. The term "anti-coagulant" shall include heparin, and warfarin. The term "thrombolytic agent" shall include agents such as streptokinase and tissue plasminogen activator.
The term "platelet anti-aggregation agent" shall include agents such as aspirin and dipyridamole.
The term "alkyl" means straight or branched alkane containing 1 to about 10 carbon atoms, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, pentyl, iso-amyl, hexy, octyl radicals and the like; the term alkenyl means straight or branched alkene containing 2 to about 10 carbon atoms, e.g., propylenyl, buten-1-yl, isobutenyl, pentenylen-1-yl, 2,2-methylbuten-lyl, 3-methylbuten-l-yl, hexen-l-yl, hepten-1-yl, and octen-1-yl radicals and the like; alkynyl means straight or branched alkyne containing 2 to about 10 carbon atoms, ethynyl, propynyl, butyn-l-yl, butyn-2-yl, WO 98/00134 PCT/US97/11133 -33 pentyn-1-yl, pentyn-2-yl, 3-methylbutyn-l-yl, hexyn-1-yl, hexyn-2-yl, hexyn-3-yl, 3,3-dimethylbutyn-1-yl radicals and the like.
The term "aryl" means a 5- or 6-membered aromatic ring containing 0, 1, or 2 heteroatoms selected from O, N, and S, e.g.
phenyl, pyridine, pyrimidine, imidazole, thiophene, oxazole, isoxazole, thiazole, and amino- and halogen- substituted derivatives thereof.
The terms "alkyloxy" or "alkoxy" include an alkyl portion where alkyl is as defined above, methyloxy, propyloxy, and butyloxy.
The terms "arylalkyl" and "alkylaryl" include an alkyl portion where alkyl is as defined above and to include an aryl portion where aryl is as defined above. The CO-n or Cl-n designation where n may be an integer from 1-10 or 2-10 respectively refers to the alkyl component of the arylalkyl or alkylaryl unit. Examples of arylalkyl include benzyl, fluorobenzyl, chlorobenzyl, phenylethyl, phenylpropyl, fluorophenylethyl, chlorophenylethyl, thienylmethyl, thienylethyl, and thienylpropyl. Examples of alkylaryl include toluene, ethylbenzene, propylbenzene, methylpyridine, ethylpyridine, propylpyridine, butylpyridine, butenylpyridine, and pentenylpyridine.
The term "halogen" includes fluorine, chlorine, iodine and bromine.
The term "oxy" means an oxygen atom. The term "thio" means a sulfur atom. Under standard nonmenclature used throughout this disclosure, the terminal portion of the designated side chain is described first followed by the adjacent functionality toward the point of attachment. For example, a C1-6 alkyl substituted with alkyl-carbonylamino is equivalent to
HO
I II Cl -6-alkyl-N-C-C In the schemes and examples below, various reagent symbols have the following meanings: WO 98/00134 PCT/US97/11133 -34-
BOC
(or Boc): Pd-C:
DMF:
DMSO:
CBZ:
CH2C12: CHC13: EtOH: MeOH: EtOAc: HOAc:
BOP:
t-butyloxycarbonyl palladium on activated carbon catalyst dimethylformamide dimethylsulfoxide carbobenzyloxy methylene chloride chloroform ethanol methanol ethyl acetate acetic acid benzotriazol-1 -yloxytris(dimethylamino)phosphonium, hexafluorophosphate 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride potassium peroxymonosulfate lithium diisopropylamide Chloro-N,N,N',N'-bis(pentamethylene)formamidinium hexafluorophosphate N-methylmorpholine The compounds of the present invention can be
EDC:
Oxone:
LDA:
PYCLU:
NMM:
administered in such oral forms as tablets, capsules (each of which includes sustained release or timed release formulations), pills, powders, granules, elixirs, tinctures, suspensions, syrups, and emulsions.
Likewise, they may be administered in intravenous (bolus or infusion), intraperitoneal, subcutaneous, or intramusculsar form, all using forms well known to those of ordinary skill in the pharmaceutical arts. An effective but non-toxic amount of the compound desired can be employed as an anti-aggregation agent.
Compounds of the invention may be administered to patients where prevention of thrombosis by inhibiting binding of fibrinogen to the platelet membrane glycoprotein complex IIb/IIIa receptor is desired. They are useful in surgery on peripheral arteries WO 98/00134 PCT/US97/11133 (arterial grafts, carotid endarterectomy) and in cardiovascular surgery where manipulation of arteries and organs, and/or the interaction of platelets with artificial surfaces, leads to platelet aggregation and consumption. The aggregated platelets may form thrombi and thromboemboli. Compounds of this invention may be administered to these surgical patients to prevent the formation of thrombi and thromboemboli.
Extracorporeal circulation is routinely used for cardiovascular surgery in order to oxygenate blood. Platelets adhere to surfaces of the extracorporeal circuit. Adhesion is dependent on the interaction between gp IIb/IIIa on the platelet membranes and fibrinogen adsorbed to the surface of the circuit. (Gluszko et al., Amer.
J. Physiol., 252(H), 615-621 (1987)). Platelets released from artificial surfaces show impaired hemostatic function. Compounds of the invention may be administered to prevent adhesion.
Other applications of these compounds include prevention of platelet thrombosis, thromboembolism and reocclusion during and after thrombolytic therapy and prevention of platelet thrombosis, thromboembolism and reocclusion after angioplasty or coronary artery bypass procedures. They may also be used to prevent myocardial infarction.
The dosage regimen utilizing the compounds of the present invention is selected in accordance with a variety of factors including type, species, age, weight, sex and medical condition of the patient; the severity of the condition to be treated; the route of administration; the renal and hepatic function of the patient; and the particular compound or salt thereof employed. An ordinarily skilled physician or veterinarian can readily determine and prescribe the effective amount of the drug required to prevent, counter, or arrest the progress of the condition.
Oral dosages of the present invention, when used for the indicated effects, will range between about 0.01 mg per kg of body weight per day (mg/kg/day) to about 100 mg/kg/day and preferably WO 98/00134 PCT/US97/11133 -36- 0.01-100 mg/kg/day and most preferably 0.01-20 mg/kg/day. For example, a typical 90 kg patient would receive oral dosages ranging between about 0.9 mg/day and about 9 g/day, most preferably between about 0.9 mg/day and 1.8 g/day. Suitable pharmaceutical oral compositions such as tablets or capsules may contain between 10-500 mg of active drug, for example, 10 mg, 100 mg, 200 mg and 500 mg.
Intravenously, the most preferred doses will range from about 1 to about 10 mg/kg/minute during a constant rate infusion.
Advantageously, compounds of the present invention may be administered in divided doses of two, three, or four times daily.
Furthermore, preferred compounds for the present invention can be administered in intranasal form via topical use of suitable intranasal vehicles, or via transdermal routes, using those forms of transdermal skin patches well known to those of ordinary skill in that art. To be administered in the form of a transdermal delivery system, the dosage administration will, or course, be continuous rather that intermittent throughout the dosage regime.
In the methods of the present invention, the compounds herein described in detail can form the active ingredient, and are typically administered in admixture with suitable pharmaceutical diluents, excipients or carriers (collectively referred to herein as "carrier" materials) suitably selected with respect to the intended form of administration, that is, oral tablets, capsules, elixirs, syrups and the like, and consistent with convention pharmaceutical practices.
For instance, for oral administration in the form of a tablet or capsule, the active drug component can be combined with an oral, non-toxic, pharmaceutically acceptable, inert carrier such as lactose, starch, sucrose, glucose, methyl cellulose, magnesium stearate, dicalcium phosphate, calcium sulfate, mannitol, sorbitol and the like; for oral administration in liquid form, the oral drug components can be combined with any oral, non-toxic, pharmaceutically acceptable inert carrier such as ethanol, glycerol, water and the like. Moreover, when desired or necessary, suitable binders, lubricants, distintegrating agents and coloring agents can also be incorporated into the mixture. Suitable WO 98/00134 PCT/US97/11133 -37binders include starch, gelatin, natural sugars such as glucose or betalactose, corn-sweeteners, natural and synthetic gums such as acacia, tragacanth or sodiumnalginate, carboxymethylcellulose, polyethylene glycol, waxes and the like. Lubricants used in these dosage forms include sodium oleate, sodium stearate, magnesium stearate, sodium benzoate, sodium acetate, sodium chloride and the like. Disintegrators include, without limitation, starch methyl cellulose, agar, bentonite, xanthan gum and the like.
WO 98/00134 WO 9800134PCrT/US97/11133 38 SCI-EME 1
H
2 N& C0 2
CH
3
HC~I-H
LI
Cl n-butanol A INHN N N
CO
2
CH
3 1-3
BOC
2 O, NEt 3
DMF
BOC-N N N 00 2
CH
3 1N NaOH/ /EtOH BOG-N \jN' C0 2
H
H
2 N Br IBOP, NMM, DMF 600 0 BO-N C-NH-- Br- 1-7
C
2 1Ve, NEt 3 o(tOlYl) 3
P,
CH
3
CN
Pd(OAC) 2 sealed tube 10000C WO 98/00134 WO 980134PrIUS97/11133 39 SCHEME 1. CONTINUED 0 Boc-N N \NIIC-NH
COM
Pd/C, H 2 EtOH 0 Boc-N N C-NH /\COM 1 iN NaOH EtOH 0 Boo- N N IC-NH \CO 2 TFA, CH 2 C1 2 0 11 02 HN N C-NH /\C 2 1 -11 WO 98/00134 PCT/US97/11133 HN N-
COCH
3 1-3 Methyl 4-(1-piperazinyl)benzoate (1-3) A solution of amine 1-IJ (20.0 g, 132 mmol), amine 1-2 (23.6 g, 132 mmol) and n-butanol (500 ml) was refluxed for 168 h.
The solution was allowed to cool to ambient temperature. The crystals were collected, washed with Et20 and dried in vacuo to give ester 1-3 as a white solid.
IH NMR (CD30D): 6 7.86 J=9Hz, 2H), 7.98 J=9Hz, 2H), 3.78 3H), 3.53 4H), 3.31 4H).
BocN N C0 2
CH
3 1-4 Methyl 1 -dimethylethoxycarbonyl)piperazin-1 -vl)benzoate (1-4) To a stirred solution of amine 1-3 (15.0 g, 61.1 mmol), NEt3 (7.42 g, 73.4 mmol) and DMF (150 ml) was added Boc20 (14.7 g, 67.2 mmol). After 1.0 h, the solution was diluted with EtOAc and then washed with H20, 10% KHSO4, brine, dried (MgSO4) and concentrated to furnish ester 1-4 as a yellow solid.
TLC Rf 0.63 (silica, 40% EtOAc/hexanes) IH NMR (CD30D): 8 7.91 J=9Hz, 2H), 7.01 J=9Hz, 2H), 3.88 3H), 3.59 4H), 3.38 4H).
WO 98/00134 WO 9800134PCT/US97/11133 -41- BocN N C0 2
H
1 -Dimethvlethoxvcarbonyl)p2iperazin- 1 -vl~benzoic acid (1 -A solution of ester14 (21.1 6 1.1 mmol) I N NaOH (100 ml, 100 mmrol) and EtOH (200 ml) was heated to 60'C for 2.0 h. The solution was acidifed with 10% KHS04 and then extracted with EtOAc.
The EtOAc phase was washed with brine, dried (MgSO4) and concentrated to furnish acid 1-5 as a white solid.
I
1 H NMR (CD3OD): 8 7.81 J=9Hz, 2H), 6.88 J=9Hz, 2H), 3.49 (in, 4H), 3.24 (in, 4H), 1.40 9H).
0 BocN \2N-0 H O
B
N-(4-bromophenyl)-4-(4-(, 1, -dimethylethoxycarbonyl)piperazin-l 1yl)benzamide A solution of acid 1-5 (400 mg, 1.31 mnmol), amine 1-6 (248 mg, 1.44 inmol), BOP reagent (867 mg, 1.97 inmol), NMM (575 kl, 5.24 inmol) and DMF was heated at 60'C for 72 h. The solution was diluted with EtOAc and then washed with H20, sat. NaHCO3, KHS04 brine, dried (MgSO4) and concentrated. Flash chromatography (silica, 20% EtOAc/hexanes) furnished amide 1-7 as a yellow solid.
TLC Rf 0.36 (silica, 20% EtOAc/hexanes) I H NMR (CDCl3): 5 7.78 J=~9Hz, 2H), 7.70 I 7.54 J=9Hz, 2H), 7.46 J=9Hz, 2H), 6.92 J=9Hz, 2H), 3.58 (in, 4H), 3.30 (in, 4H), 1.49 9H).
WO 98/00134 WO 9800134PCTIUS97/11133 42 BocN N Q NH (9
.CO
2 Me (E)-Methyl 1,1 -diniethylethoxycarbonyl)piperazin- 1vl)p2henylcarbonylaminophenyl)p2rop-3 -enoate (1-8) A solution of amnide 15-7 (300 mg, 0.6519 mmol), methyl acrylate (587 g~l, 6.52 mmol), 0(tOlyl)3P (120 mg, 0.311 mmol), NEt3 (184 p1, 1.30 mmol), Pd (OAc)2 (15 mg, 0.05 12 minol) and CH3CN ml) was heated to 100 0 C in a sealed tube for 18 h. The solution was diluted with EtOAc, and then washed with H20, sat. NaHCO3, KHSO4, brine, dried (MgSO4) and concentrated. Flash chromatography (silica, 25% 4 40% EtOAc/hexanes) furnished olefin 1-8 as a yellow solid.
TLC Rf 0.36 (silica, 50% EtOAc/hexanes) IH NMR (CDCl3): 8 7.87 1H), 7.81 J=9Hz, 2H), 7.68 (in, 3H), 7.53 J=-9Hz, 2H), 6.94 J=9Hz, 2H), 6.39 J=l6Hz, 1H), 3.81 (s, 3H), 3.61 (mn, 4H), 3.30 (in, 4H), 1.50 9H).
Bc\ N0 -NH-O
CO
2 Me Methyl 1,1-dimethylethoxycarbonyl)piperazin- 1yl)p2henylcarbonylaminophenyl)prop~anoate Q1-9) A mixture of ester 1-8 (260 mg, 0.56 mmnol), 10% Pd/C (100 mng) and EtOH (10 ml) was stirred under I atm H2 for 18 h. The reaction mixture was filtered through a celite pad and concentrated to furnish ester 1-9 as a brown solid.
I H NMR (CDC13): 8 7.79 J=9Hz, 2H), 7.67 lH), 7.54 1=8Hz, 2H), 7.19 J=8Hz, 2H1), 6.92 J=9Hz, 2H), 3.67 3H), 3.60 (in, WO 98/00134 PCT/US97/11133 -43- 4H), 3.29 4H), 2.94 J=8Hz, 2H), 2.63 J=8Hz, 2H), 1.49 (s, 9H).
0 BocN N C-NH
CO
2
H
1-10 -dimethylethoxycarbonyl)piperazin- 1yl)phenlcarbonylaminophenyl)propanoic acid (1-10) A solution of ester 1-9 (250 mg, 0.53 mmol), 1 N NaOH (1 ml, 1.00 mimol) and EtOH (3 ml) was stirred at ambient temperature for h. The solution was acidified with 10% KHSO4 and then extracted with EtOAc. The EtOAc phase was washed with brine, dried (MgSO4) and concentrated to furnish acid 1-10 as a tan solid.
TLC Rf 0.11 (silica, 10:0.2:0.2 CH2Cl2/MeOH/AcOH) 1H NMR (CD30D): 6 7.86 J=9-Hz, 2H), 7.55 J=8Hz, 2H), 7.20 J=9Hz, 2H), 7.08 J=8Hz, 2H), 3.60 (bs, 41-H) 3.45 4H), 2.89 J=8Hz, 2H), 2.59 J=8Hz, 2H), 1.48 9H).
0
II
HN N -(C-NH
CO
2
H
1-11 3-(4-(4-piperazin-1 -ylphenylcarbonylamino)phenyl) propanoic acid (1-11) To a solution of acid 1-10 (215 mg, 0.4743 mmol), TFA mi) and CH2Cl2 (3.0 ml) was stirred at ambient temperature for h. The solution was concentrated and then azeotroped with toluene.
The residue was trituated with 10:0.2:0.2 EtOH/NH40H/H20, filtered, washed with Et20 and dried in vacuo to furnish acid 1-11 as a tan solid.
WO 98/00134 PCTIUS97/11 133 -44- TLC Rf 0.55 (silica, 10: 1: 1 EtOH/NH4OH/H20) I H NMR (D20): 8 7.83 J=9Hz, 2H), 7.42 J=8Hz, 211), 7.31 (d, J=±SHz, 2H), 7.16 (d,-J=9Hz, 2H), 3.25 (in, 4H), 2.96 (in, 4H, 2.89 (t, J=-8Hz, 2H), 2.49 J=8Hz, 2H).
WO 98/00134 WO 9800134PCTfUS97/11133 45 SCHEME 2 BOCN
CO
2 H HCI -H 2 N O 2-2 BOP, NMM
DMF
600 0 BOO-N \2N C-H OH Br II CO 2 tBu
CS
2 00 3
DMF
BOO-N C-NH-& \t J T A, CH 2 1 2 0 HN C0 2
H
2-6 WO 98/00134 PCT/US97/11133 46 BocN N NHOH N-(4-Hydroxyphenyl)-4-(4-( 1,1 -dimethylethoxycarbonyl)piperazinyll -yl)benzamide (2-3) A solution of carboxylic acid 2-1 (400 mg, 1.31 mmole), 4aminophenyl 2-2 (210 mg, 1.44 mmol), BOP reagent (867 mg, 1.97 mmole), NMM (575 pl1, 5.24 mmol) and DMF (10 mi) was heated to 0 C for 72 h. The reaction mixture was diluted with EtOAc and then washed with H20, sat NaHCO3, 10% KH-1SO4, dried (MgSO4) and then concentrated. Flash chromatography (silica, 50% EtOAc/hexanes) furnished the amide 2-3 as a white solid.
TLC Rf 0.25 (silica, 50% EtOAc/hexanes) H NMR (CD30D): 6 7.82 J=9Hz, 2H), 7.40 J=9Hz, 2H), 7.0 (d, J=9Hz, 2H), 6.75 J=9Hz, 2H), 3.57 (bs, 4H), 3.30 (bs, 4S), 1.47 (s, 9H).
0 BocN N OC H 2
CO
2 tBu tert-Butyl 2-(4-(4-(4-(1,1-dimethylethoxycarbonyl)piperazinyl- 1vl)phenvlcarbonylamino)phenoxy)acetate To a stirred solution of amide 2-3 (400 mg, 1.01 mmol) and DMF (5 ml) was added cesium carbonate (820 mg, 2.53 mmole) followed by t-butyl bromoacetate (195 p1, 1.21 mmol). After 20 h, the reaction mixture was diluted with EtOAc and then washed with 10% KHSO4, brine, dried (MgSO4) and concentrated. Flash chromatography (silica, 25% EtOAc/hexanes) furnished ester 2-5 as a white solid.
WO 98/00134 WO 9800134PCT/US97/11 133 47 TLC Rf 0.40 (silica, 50% EtOAc/hexanes) I H NMR (CDCI3): 5 7.79 J=9Hz, 2H), 7.63 1 7.54 J=9Hz, 2H), 6.92 (in, 4H4), 4.51 2H), 3.60 (in, 4H), 3.30 (mn, 4H), 1.50 (s, 18H).
0 HN CNH a OOH 2
CO
2
H
2-(4-(4-Piperazinyl -1 -yl)phenylcarbonylamino)phenoxy) acetic acid (2-6) A solution of ester 2- (275 mng, 0.5378 mmol), TFA ml) and CH2Cl2 (5 ml) was stirred at ambient temperature for 2.0 h.
The solution was concentrated and then azeotroped with toluene. The residue was trituated with 10:0.5:0.5 EtOH/NI-40H/H20, filtered, washed with EtOH and then washed with EtOH to furnish acid 2-6 as a white solid.
TLC Rf 0.34 (silica, 10:0.5:0.5 EtOHINI-40H/H20) I H NMR (CD3 OD): 8 7.8 3 J=9Hz, 2H), 7.50 J=9Hz, 2H), 7 .00 J=9Hz, 2H), 6.92 J=9Hz, 2H), 4.36 2H), 3.30 (in, 4H), 2.96 (mn, 4H).
WO 98/00134 WO 9800134PCT[US97/11133 48 SCHEME 3 0 BOO-N N C-NH-- oC2B
CH
3
I,
DMF,
CS
2 00 3 600C 0 BOO-N N C-N 0 CO 2 tBu
OH
3 3-1.
ITFA, CH 2 C1 2 0 HN N C-N 0 C0 2
H
O2 H 3 WO 98/00134 WO 9800134PCTIUS97/11 133 49 0 BocN N C -N OCH 2 00 2 tBu 3-1 CH 3 tert-Butyl 1,1-dimethylethoxycarbonyl)piperazin- Iyl)p2henylcarbonyl(N-methyl)amino)p2henoxv')acetate (3-1) To a stirred solution of ester 2-5 (200 mg, 0.3911 mmol) was added cesium carbonate (318 mg, 0.9778 mmole) followed by CH31 (29 gl, 0.47 mmol). After heating at 600 for 2.0 h, the reaction mixture was diluted with EtOAc and then washed with H20, 10% KHSO4, brine, dried (MgSO4) and concentrated. Flash chromatography (silica, EtOAc/hexanes) furnished ester 3-1 as a yellow solid.
TLC Rf 0.45 (silica, 50% EtOAc/hexanes) I H NMR (CDC13): 5 7.21 J=-9Hz, 2H), 6.97 J=9Hz, 2H), 6.75 (d, J=9Hz, 2H), 6.63 J=9Hz, 2H), 4.45 2H), 3.52 (bt, 4H), 3.43 (s, 3H), 3.13 (bt, 4H), 1.47 18H).
0 HN N C -N E OCH 2 00 2
H
32CH 3 -Piperazinyl)phenylcarbonyl(N-methyl)amino)phenoxy)-acetic acid (3-2) A solution of ester 3-1. (190 mg, 0.36 17 mmole), TFA (3 ml) and CH2C12 (3 ml) was stirred at ambient temperature for minutes. The solution was concentrated and then azeotroped with toluene. Flash chromatography (silica, 10:0.2:0.2 EtOH/N1-40H/H20) furnished acid 3-2 as a white solid.
TLC Rf 0.42 (silica, 10:0.2:0.2 EtOH/NH4OH/H20) WO 98/00134 WO 9800134PCT/US97/1 1133 50 H NMR (CD3 OD): 8 7.25 J=9Hz, 2H), 7.06 J=9Hz, 2H), 6.91 J=9Hz, 2H), 6.82 J=9Hz, 2H), 4.66 2H), 3.50 (in, 4H), 3.41 (s, 3H), 3.35 (mn, 4H).
SCHEME 4
HN
1 C NH -0 O C 2
H
HCI (g) EtOH, 000 RT 0 HCI.HN N& C-NH 0 "CO 2
CH
2
CH
3 4-1 Ethyl -piperazinyl)phenylcarbonylamino)phenoxy)acetate hydrochloride To a stirred solution of acid 2-6 (40 mg, 0.1125 inmol) and EtOH (2 ml) at 0 0 C was bubbled HCI gas for 3 minutes. After 24 h at ambient temperature, the solution was concentrated to give ethyl ester 4- 1 as a white solid.
1 H NMR (D20): 8 7.87 J=9Hz, 2H), 7.44 J=9Hz, 2H), 7.19 (d, J=9Hz, 2H), 7.05 J=9Hz, 2H), 4.80 2H), 4.31 J=7Hz, 2H) 3.62 (in, 4H), 3.43 (mn, 4H), 1.30 J=7Hz, 3H).
WO 98/00134 WO 9800134PCT/US97/11133 51 SCHEME Bo N \N-C2 2- 5-1 BOP, NMM
OME
0 11 H Boc-N NC-NH Br-ThO 2 tBu 5-3 SNaH, THF 0 /I I Boc-N N' -NH N'Ctu 5-4 TFA, CH 2
CI
2 0 HN N -j NC-NH 1
N/'CO
2
H
WO 98/00134 WO 9800134PCTIUS97/11 133 52 BocN N CN CNH
H
5-2 N-(5-Lndolyl)-4-(4-( 1,1 -dimethylethoxycarbonyI)piperazin- 1vl~benzamide (5-2) To a stirred solution of acid 2- (500 mg, 1.63 mmol), NMM (715 gi, 6.52 mmol) and DMF (5 ml) was added BOP reagent (1.08 g, 2.45 mmol). After 30 minutes, amine 5-1. (Aldrich 260 mg, 1.96 mmole) was added. After 18 h at 60'C, the solution was diluted with EtOAc and then washed with H20, brine, dried (MgSO4) and concentrated. Flash chromatography (silica, 20% EtOAc/CHC13) afforded amide 5-2 as a yellow solid.
TLC Rf 0.13 (silica, 20% EtOAc/CHCl3) IH NMR (10% CD3OD/CDCl3): 567.90 1H), 7.83 J=9Hz, 2H), 7.63 (in, 2H), 7.22 (9s, 1H), 6.94 J=9Hz, 2H), 6.52 J=3Hz, lH), 3.59 (in, 4H), 3.28 (in, 4H), 1.49 9H).
0 BOC-N N O -C-N .H N/C~ 5-4 tert-Butyl 1,1-dimethylethoxycarbonyl)piperazin- 1vl)phenyl carbonylamino)indol- I -vl acetate (5-4) To a stirred solution of amide 5-2 (300 mg, 0.71 mmol) and THF (5 ml) was added NaR (60% dispersion in mineral oil; 29 mg, 0.7137 rnrol). After 10 minutes, bromide 5-3 '(115 gl, 0.71 mmol) was added. After 30-minutes, the solution was diluted with EtOAc and then washed with H2Q, brine, dried (MgSO4) and concentrated. Flash WO 98/00134 PCT/US97/11133 -53 chromatography (silica, 40% EtOAc/hexanes) afforded ester 5-4 as a yellow solid.
TLC Rf 0.18 (silica, 40% EtOAc/hexanes) 1H NMR: 5 7.92 1H), 7.83 J=9Hz, 2H), 7.77 IH), 7.39 (dd, J=9Hz, 2Hz, 1H), 6.93 J=9Hz, 2H), 6.52 J=3Hz, 1H), 4.72 (s, 2H), 3.60 4H), 3.28 4H), 1.49 9H), 1.43 9H).
0 /1 CO, IH HN N -NH N CO2 2-(5-(4-(1-Piperazinyl)phenylcarbonylamino)indol-1 -yl) acetic acid To a stirred solution of ester 5-2 (250 mg, 0.4678 mmol), anisole (203 gl, 1.87 mmol) and CH2C12 (3 ml) at 0°C was added TFA.
After 30 min., the solution was concentrated and then azeotroped with toluene. Flash chromatography (silica, 10:0.1:0.1 EtOH/NH40/H20) afforded crude acid. The crude material was purified by prep HPLC to furnish acid TLC Rf 0.31 (silica, 10:0.5:0.5 EtOH/NH40H/H20) 1 H NMR (50:50 d6- DMSO/D20, 2 drops IN NaOD): 5 7.87 (d, J=9Hz, 2H), 7.71 1H), 7.24 3H), 7.07 J=9Hz, 2H), 6.47 (d, J=3Hz, 1H), 4.63 2H), 3.25 4H), 2.89 4H).
WO 98/00134 WO 9800134PCT/US97/11133 54 SCHEME 6 Boo-N C0 2
H
Br
INH
2 BOP, NMM, DMF, 0 Boo- N N N H 6-2 E 2 Me, NEt 3 o(toiYl) 3 p, CH 3
CN,
Pd(OAC) 2 sealed tube 1 00 0
C
0 N CO 2 Me Boc-N N C-NH Pd/C EtOH,
H
2 Boo- N N /\CN 6-4 1N NaOH EtOH WO 98/00134 PCTIUS97/11133 55 SCHEME 6 CONTINUED o C0 2
H
Boc-N NCNH ITFA,
CH
2
CI
2 o
CO
2
H
HN N
N
6-6 BocN ~N
CN
k-I4 fi:2Br N-(3-bromophenyl)-4-(4-(, 1, -dimethylethoxycarbonyl)piperazin.I 1yl))benzamide (6-2) A solution of acid 1-5~ (400 mg, 1.31 mimol), amine 6-1 (248 mg, 1.44 rnmol), BOP reagent (867 mg, 1.97 mmol), NMM (575 gil, 5.24 mmol) and DMF (10 ml) was heated to 60'C for 72 h. The solution was diluted with EtOAc and then washed with H20, sat.
NaHCO3, 10% KHSO4, brine, dried (MgSO4) and concentrated. Flash chromatography (silica, 25% EtOAc/hexanes) furnished amide 6-2 as a yellow solid.
TLC Rf 0.31 (silica, 25% EtOAc/hexanes) I H NMR (CD3 OD): 8 7.99 I 7.84 J=9Hz, 2H), 7.60 (in, IlH), 7.25 (in, 2H), 7.02 J=9Hz, 211), 3.57 (mn, 4H), 3.31 (mn, 4H), 1.47 (s, 9H).
WO 98/00134 PCTIUS97/11133 56
CO
2 Me 0 Boc:N N CNH K (-Ej-Methyl (3 1,1-dimethylethoxycarbonyl)piperazin yl))phenyl)carbonvaminophenl)prop-3-.enoate (6-3) A solution of arnide 6- (400 mg, 0.86 mmol), methyl acrylate (783 p1l, 8.69 mmole), O(tolyl)3P (159 mg, 0.415 mmol), NEt3 (245 gl, 1.74 mmole), Pd(OAc)2 (20 mg, 0.069 mmole) and CH3CN ml) was heated to 100 0 C in a sealed tube for 18 h. The solution was diluted with EtOAc and then washed with H20, sat. NaHCO3, KHSO4, brine, dried (MgSO4) and concentrated. Flash chromatography (silica, 20% EtOAc/hexanes) furnished olefin 6-3 as a yellow solid.
TLC Rf 0.47 (silica, 50% EtOAc/hexanes) 1 H NMR (CD3 OD): 5 7.94 I 7.87 (dd, J=2Hz, 9Hz, 2H), 7.71 (in, 2H), 7.36 (in, 2H), 7.03 (dd, J=2Hz, 9Hz, 2H), 6.52 J=l6Hz, 3.78 3H), 3.57 (mn, 4H), 3.29 (in, 4H), 1.47 9H).
o
CO
2 Me BocN N -OC-NH Methyl 1,1-dimethylethoxycarbonyl)piperaziny l)Phenyl) carbon y Iaminophenyl)propaoate (6-4) A mixture of ester 6- (300 mg, 0.64 mmol), 10% Pd/C (100 mg) and EtOH (10 ml) was stirred under I atm H2 for 18 h. The WO 98/00134 PCT/US97/11133 57 reaction mixture was filtered through a celite pad and concentrated to furnish ester 6-4 as a yellow oil.
I H NMR (CDCI3): b 7.80 (in, 3H), 7.54 I 7.47 J-8Hz, I H), 7.27 (in, 1H), 6.96 8Hz, 11H), 6.92 J=9Hz, 2H), 3.68 3.60 (in, 4H), 3.29 (in, 4H), 2.96 J=8Hz, 2H), 2.66 J=8Hz, 2H), 1.50 (s, 9H).
o
CO
2
H.
BOO-N C-NH
K
3 1,1 -Dimethylethoxycarbonyl)piperazin.1 -yl))phenyl)carbonylaminophenyrpr-opanoic acid A solution of ester 6-4 (260 mg, 0.5563 minol), 1 N NaOH (1 ml, 1 mmol) and EtOI- (3 ml) was stirred at ambient temperature for h. The solution was acidified with 10% KHS04 and then extracted with EtOAc. The EtOAc phase was washed with brine, dried (MgSO4) and concentrated to furnish acid 6-5 as a white solid.
TLC Rf 0.08 (silica, 10:0.2:0.2 CH2C12IMeOH/IAcOH) I H NMR (CD3OD): 6 7.84 J=9Hz, 2H), 7.51 (in, 2H), 7.25 (t, J=8Hz, 1H), 7.03 (mn, 3H), 3.57 (in, 411), 3.29 (mn, 4H), 2.91 J=8Hz, 2H), 2.60 J=8Hz, 2H1), 1.47 9H).
WO 98/00134 WO 9800134PCTIUS97/11133 58 0 C0 2
H
II 7 3 -(3-(4-Piperazin- 1 -ylphenyl)carbonylamino)phenyl) p2rop~anoic acid (6-6) A solution of acid 6- (210 mg, 0.4633 mmol), TFA (3 ml) and CH2Cl2 (3 ml) was stirred at ambient temperature for 1.0 h. The solution was concentrated and then azeotroped with toluene. Flash chromatography (silica, 10:0.1:0.1 -4 10:0.5:0.5 EtOHINII4OH/1- 2 0) furnished acid 6-6 as a tan solid.
TLC Rf 0.59 (silica, 10: 1: 1 EtOHINH4OH/H20) I H NMR (D20): 8 7.82 J=9Hz, 2H), 7.38 (in, 2H), 7.34 I1H), 7.15 J=9Hz, 3H), 3.25 (in, 4H), 2.96 (in, 4H), 2.90 J=8Hz, 2H), 2.50 J=8Hz, 2H).
WO 98/00134 WO 9800134PCT/US97/11 133 59 SCHEM 7 I-0 BocN
OH
3 2-3 B r7-1CO 2 Et
ICS
2 C0 3
DMF
TFA, CH 2
CI
2 NaOH, EtOH HN t C0 2
H
WO 98/00134 WO 9800134PCT/UJS97/11133 BocN N Q
CH
3 NH (Q 0CO 2 Et Ethyl ,1 -Dimethylethoxycarbonyl)piperazin- 1vi )phenylcarbonylamino )phenoxypropanoate (7-2) Phenol 2- (381 mg, 0.993 nimol), ethyl 2-bromopropionate 129 gL, 0.99 mmol), and Cs2CO3 (807 mg, 2.5 nunol) were combined in 5 mL DMF. After stirring overnight the reaction was diluted with EtOAc, washed with water 10% KHS04 (3x) and brine, dried (MgSO4), filtered and concentrated. Flash chromatography (silica, 40% EtOAc/hexane) provided 7-2 as a white solid.
TLC Rf 0.41 (silica, 50% EtOAc/hexane) I H NMR (400 MHz, CDCl3): 8 7.78 J=9Hz, 2H), 7.60 1H), 7.52 J=9Hz, 2H), 6.92 J=9Hz, 2H), 6.89 J=9Hz, 2H), 4.72 (q, J=7Hz, IH), 4.22 J=7Hz, 2H), 3.59 (in, 4H), 3.28 (in, 4H), 1.61 (d, J=7Hz, 3H), 1.49 9H), 1.26 J=7Hz, 3H).
HN N 0
H
3 NH Q C<0 2 Et Ethyl 2-(4-(4-(piperazin- 1-yl)phenylcarbonylamino) p2henoxy)propanoate(7-3) A solution of 7-2 (266 mg, 0.53 mrnol) and anisole (200 g.L, 1.8 minol) in 5 mL 1: 1 CH2Cl2,TFA was stirred for 20 min. After concentration and azeotroping with toluene, flash chromatography (silica, 33:1-:1 EtOHIH2O/NH4OH) provided 7-3 as a white solid.
TLC Rf =0.37 (silica, 20: 1:1 EtOH/H2OINII4OH WO 98/00134 WO 98/0 134PCT/US97/11133 61 I H NMR (400 MHz, CD3OD): 5 7.84 J=9Hz, 2H), 7.54 J=9Hz, 2H), 7.01 J=9Hz, 2H), 6.88 J=9Hz, 2H), 4.21 J=7Hz, 2H), 3.31 (in, 4H), 2.98 (rn, 4H), 1.57 J=7H1z, 3H), 1.26 J=Hz, 3H).
HN 0 CH 3 NH Q .OC0 2
H
7-4 2-(4-(4-(Piperazin- 1 -yl)phenylcarbonylamino)phenoxy) p2rop~ionic acid (7-4h- Ester 7-3 (44 mg, 0.11 rnmol) was dissolved in 1 mL EtOH, then 1 M NaOH (122 jiL, 0. 12 minol) was added. After 3 h the reaction was neutralized with I N HCl, concentrated, then purified by flash chromatography (silica, 50:1:1 EtOH/H2OINH4OH) providing 7-4 as a white solid.
TLC Rf 15 (silica, 50: 1:1 EtOH/H2OINI-40H) 1 H NMR (400 MHz, D20): 867.69 J=9Hz, 2H), 7.22 J=9H, 2H), 7.01 J=9Hz, 2H), 6.80 J=9Hz, 2H), 4.48 J--7Hz, IH), 3.10 (in, 4H), 2.82 (mn, 4H), 1.40 J=7Hz, 3H).
WO 98/00134 WO 9800134PCTIUS97/11133 62 SCHEME 8 cl- CO 2 Et 1) Piperazine, NMP, 500T 2) Boc 2
O
BocN N N- CO 2 Et NaOH, EtOH BocN N BoN N N
O
H tBUtyl bromoacetate C0 3
DMF
1) BOP Reagent, DMF 2) 4-aminophenol, 6000C BooN N 0 Boc/ N,,-C0 2 -t-Bu N- 0 L H.- /TEA, OCH 2 C12 HN N 0 0 H-6q0 2
H
WO 98/00134 PCT/US97/11133 -63 BocN N CO 2 Et 8-2 Ethyl 6-r4-(1.1 -dimethvlethoxvcarbonyl)piperazin-l-vllnicotinate (8-2) Ethyl 6-chloronicotinate Maybridge Chemical Co., 2.0 g, 10.8 mmol) and piperazine (1.4 g, 15 mmol) were combined in 54 mL NMP and heated at 50 0 C overnight. After cooling, Boc20 (2.6 g, 11.9 mmol) was added. The reaction mixture was stirred overnight then diluted with EtOAc, washed with water, sat NaHCO3 and brine, dried (MgSO4), filtered and concentrated. Flash chromatography (silica, 25% EtOAc/hexane) provided 8-2 as a white solid.
TLC Rf 0.63 (silica, 40% EtOAc/hexane) IH NMR (400 MHz, CDC13): 8 8.80 J=2Hz, 1H), 8.04 (dd, J=9, 2Hz, 1H), 6.58 J=9Hz, 1H), 4.34 J=7Hz, 2H), 3.68 4H), 3.54 4H), 1.49 9H), 1.37 J=7Hz, 3H).
BocN N CO 2
H
NO:
6-[4-(1,1-Dimethylethoxvcarbonyl)piperazin-1 -yllnicotinic acid (8-3) Ester 8-2 (3.0 g, 9.3 mmol) was dissolved in 93 mL EtOH, 1 M NaOH (23 mL, 23 mmol) was added, and the reaction mixture was stirred overnight. After concentrating, the residue was dissolved in water, washed with EtOAc, and the aqueous layer was acidified with KHS04. After extracting with EtOAc, the organic layer was washed with brine, dried (MgSO4), filtered and concentrated providing 8-3 as a white solid.
1H NMR (400 MHz, CD3OD): 8 8.71 J=2Hz, 1H), 8.08 (dd, J=9, 2Hz, 1H), 6.77 J=9Hz, IH), 3.61 4H), 3.53 4H), 1.49 (s, 9H).
WO 98/00134 WO 9800134PCTfUS97/11133 64 BocN N N-(4-Hydroxyphenyl)-6-(4-(, 1, -dimethylethoxycarbonyl)piperazin- 1 vlhnicotinamide (8-4) Acid 8-3 (500 mg, 1.6 mmol), NMM (720 jiL, 6.5 mmol) and BOP reagent (793 mg, 1.8 mmol) were combined in 8 mL DMF.
After 1.5 h, 4-aminophenol (260 mg, 1.8 mimol) was added and the reaction was heated to 60'C overnight. The reaction mixture was diluted with EtOAc, washed with water sat. NaHCO3 and brine, dried (MgSO4), filtered and concentrated. Flash chromatography (silica, 50-60% EtOAc/hexane) provided 8-4 as a brown solid.
TLC Rf 0.40 (silica, 70% EtOAc/hexane) I H NMR (400 MHz, CDCl3): 8 8.66 J=-2Hz, 1H), 7.98-7.95 (in, 2H), 7.33 J=9Hz, 2H), 6.75 J=9Hz, 2H), 6.58 J=9Hz, 1H), 3.61 (in, 4H), 3.52 (in, 4H), 1.48 (in, 9H).
C>0 BocN N4 NDI OCH 2 CO~tu tert-Butyl 1,1 -dimethylethoxycarbonyl)piperazin- 1yl)p2yridin -5-vl)carbonylamino)p2henoxyacetate Phenol 8-4 (374 mng, 0.94 mmnol), t-butyl bromoacetate (151 IlL, 0.94 inmol) and CS2CO3 (761 mng, 2.3 mmnol) were combined in 5 mE DMF. After 90 min. the reaction mixture was diluted with EtOAc, washed with water (4x) and brine, dried (MgSO4), filtered and concentrated. Flash chromatography (silica, 50% EtOAc/hexane) provided 8-5 as a brown solid.
TLC Rf 0.53 (silica, 70% EtOAc/hexane) WO 98/00134 WO 9800134PCTIUS97/1 1133 65 I H NMR (400 MI-z, CDC13): 5 8.66 J=2Hz, I 7.99 (dd, J=9, 2Hz, I1H), 7.56 I1H), 7.52 J=8I-z, 2H), 6.90 J=8Hz, 211), 6.65 J=9Hz, 1H), 4.5 1.4s, 2H), 3.68 (in, 4H), 3.56 (in, 4H), 1.49 18H).
HN N (j B& NNH Q OCH 2
CO
2
H
2-(4-(((2-Piperazin- 1 -yl)pyridin-5 -yl)carbonyl amino)phenoxy)acetic acid (8-6) Ester 8-5 (290 mg, 0.57 minol) was dissolved in 5 mL 1:1 TFA/CH2C12. After I h the reaction was concentrated and azeotroped with toluene. Flash chromatography (silica, 10: 1: 1 EtOH/H2OINH4OH) and trituration with Et2O provided 8-6 as a light brown solid.
1 H NMR (400 MHz, D20): 8 8.42 J=2Hz, I1H), 7.89 (dd, J=9, 2Hz, I1H), 7.04 J=8 Hz, 2H), 6.78 J=9Hz, 2H), 6.76 J= 10 Hz, ILH), 4.32 2H), 3.37 (in, 4H), 2.74 (in, 4H).
WO 98/00134 WO 9800134PCTIUS97/11133 66 SCHEME 9 Br &NHNH 2 0
H
3 C y 0 ,Et H_ CH 3 Br 'NN N= Et I 0 9-1
PPA
Brl(C I \CO 2 Et
SN
H~a/M
CICH
2
CN
CO
2 Et
~CN
LAH Et 2
O
9-3 WO 98/00134 WO 9800134PCTIUS97/11133 67 SCHEME 9 (CONT'D) K>l HN-
IBOC
2 0 TEA K'N,4 Br
BOON
Pd(OAC) 2 dppp/ DMF 00(g) CH: 3
OH
9-6 LiOH
THF/H
2 0/MeOH (NN ~C 2
H
BOON: j
H
2 N- D O...CO 2 Et HCI OH 3 10-4 PYCLU/i-Pr 2 NEt WO 98/00134 WO 9800134PCTIUS97/11 133 68 SCHEME 9 (CONT'D'
BOON
%,\CO
2 Et 9-8
OH
3
LIOH
TH F/H 2 0/MeOH HO /EtOAC 0 HN 2
H
CH
3 WO 98/00134 WO 9800134PCTIUS97/1 1133 69 SCHEME 9 (CONT'D) f N /\CO 2
CH
3
BOON:
NaBH 3 ON EtOAc 00 2 0H 3 9-10 I LOH IMeOH I H 2 0/THF
~/C
2
H
BOON:LHMe 9-11 WO 98/00134 WO 9800134PCTIIJS97/11133 70 SCHEME 9 (CONT'D)
H
2 N Q Ol---CO 2 Et PYCLU/i-.Pr 2 NEt HCI* H 3 10-4
BOON.
O\/COQ
2 Et 9-12
OH
3 TH F/H 2 0/MeOH IHOI/EtOAC
O\CO
2
H
9-13
OH
3 WO 98/00134 PCT/US97/11133 -71 H H 3 -1 Br N-N= Et 0 Ethyl 2-(4-bromo-1-hydrazinimine)propanoate 9-1 A mixture of 4-bromophenylhydrazine (Aldrich, 0.5 g, 2.2 mmol) and ethyl acetoacetate (Aldrich, 0.24 mL, 2.2 mmol) in pyridine (0.6 mL) was heated to reflux overnight. The reaction was cooled, diluted with water and the precipitate that resulted was collected and washed with water, dried under vacuum to give 9-1.
MeOH/CHCl3 saturated with NH3)=0.86 IH NMR (400 MHz, CDC13) 6 7.64 1H), 7.41 2H), 7.39 2H), 4.30-4.34 2H), 2.10 3H), 1.36 3H).
Br CO 2 Et
N
H 9-2 5-Bromo-2-ethoxycarbonyl indole 9-2 A mixture of 9-1 (0.54 g, 1.9 mmol) and polyphosphoric acid (1.6 mL) was heated to 115°C for 10 minutes, then diluted with cold water and extracted with EtOAc. The layers were separated and the aqueous layer extracted with EtOAc. The organic layers were combined, washed with brine, dried with MgSO4, filtered and concentrated to give 9-2 as a brown solid.
EtOAc/hexanes)=0.45 1H NMR (400 MHz, CDC13) 8 8.95(bs, 1H), 7.82 1H), 7.41 IH), 7.30 1H), 7.15 1H), 4.44 2H), 1.40 3H).
WO 98/00134 PCT/US97/11133 -72- BrI CO 2 Et N 9-3
-CN
1-(Cyanomethvl)-2-ethoxycarbonyl-5-bromo-indole 9-3 A solution of 9-2 (11.2 g, 44.4 mmol) in DMF (400 mL) was treated with NaH (3.2 g of 60% dispersion in oil, 66.6 mmol) for hour and then chloroacetonitrile (Aldrich, 5.6 mL, 88.8 mmol) was added and the reaction was stirred overnight. The solvent was removed in vacuo and the residue was partitioned between water and EtOAc.
The water layer was extracted with EtOAc, the organic layers were combined, washed with water, brine, dried with MgSO4, filtered and evaporated to give 9-3 as a brown solid.
EtOAc/hexanes)=0.46 1H NMR (400 MHz, CDC13) 8 7.82-7.83 (bs, 1H), 7.51-7.54 (dd, IH), 7.30-7.32 (bd, 2H), 5.60 2H), 4.41-4.43 2H), 1.41-1.43 3H).
B 9-4 rN NH 8-Bromo-2.3.4.5-tetrahydropyrazino-r 1,2-alindole 9-4 A slurry of 9-3 (12.2 g, 39.7 mmol) in diethyl ether (400 mL) was added via dropping funnel to a solution of LAH in ether (79.4 mL, 1 M in ether, 79.4 mmol) and stirred at room temperature overnight. The slurry was diluted with saturated sodium potassium tartrate (Rochelle's salt) and stirred for 15 minutes, then transferred to a separatory funnel containing EtOAc and the layers separated. The aqueous layer was extracted with EtOAc, the organic layers were combined, washed with water, brine, dried with MgSO4, filtered and evaporated to give 9-4 as a brown solid.
WO 98/00134 WO 9800134PCT/US97/11133 73 Rf(1O% MeOH/CHCI3 saturated with NHF3)=0.42 I H NMR (400 MHz, CDC13) 8 8.38-8.41 (bs, IlH), 7.66 I1H), 7 .2 1 7.22 (dd, 2H), 7.14 (ci, 2H), 6.14 1H), 4.22 2H), 3.99 2H), 3.36-3.37 2H).
K'N Br9-
BOON
8 -Brom o-3 -dimethylethoxycarbonyl)-2, 3,4,5 -tetrahydrop2yrazino-[1 .2-alindole A solution of 9-4 (10 g, 40 mmol) in CH2Cl2 (200 m.L) was cooled to 0 0 C and treated with di-tertbutyldicarbonate (8.7 g, mmol) and triethylamine (5.6 miL, 40 mmol). The solution was allowed to warmi slowly and after 48 hours was concentrated and the residue dissolved in EtOAc, washed with water and brine, dried over Na2SO4, filtered and evaporated. The residue was chromatographed EtOAc/hexanes) to give 9-5 as a solid.
EtOAc/hexanes)=0.22 I H NMR (400 MHz, CDCl3) 8 7.67 (di, I 7.23 IlH), 7.13 IlH), 6.21 1H), 4.80 2H), 4.04 2H), 3.93 2H), 1.50 9H).
r N 00 2
CH
3 9-6 BOON 8-Methoxycarbonyl-3-( 1,1 -dimethylethoxycarbonyl)-2,3 tetrahydropvrazino-[ 1.2-alindole 9-6 A solution of 9-5 (3.0 g, 8.5 mnmol) in MeOH (60 mL) and (20 m.L) was treated with triethylamine (3.55 m.L, 25.5 rnmol), l,3-Bis(diphenylphosphino)propane (1.75 g, 4.25 mmol) and palladium (HI) acetate (0.952 g, 4.25 mmol). Carbon monoxide was bubbled WO 98/00134 PCT/US97/11133 -74through the solution while it was heated to reflux for 2 hours. The reaction was heated at reflux overnight under a balloon atmosphere of carbon monoxide. Additional 1,3-Bis(diphenylphosphino)propane (0.8 g, 2.12 mmol) and palladium acetate (0.476 g, 2.12 mmol) were added and the reaction was heated at reflux for 48 hours under a balloon atmosphere of carbon monoxide. The reaction was cooled to room temperature, the residue was partitioned between water and EtOAc.
The water layer was extracted with EtOAc, the organic layers were combined, washed with water, brine, dried with MgSO4, filtered and evaporated. The residue was chromatographed'(25% EtOAc/hexanes) to give 9-6 as a yellow solid.
EtOAc/hexanes)=0.21 1H NMR (400 MHz, CDC13) 8 8.32 1H), 7.90 1H), 7.26 1H), 6.38 1H), 4.83 2H), 4.12 2H), 3.96-3.93 5H), 1.50 9H).
r N 'CO 2 H 9-7
BOCN
3-(1,1 -dimethylethoxycarbonyl)-2,3,4,5-tetrahydropyrazino-[ 1,2alindole-8-carboxvlic acid 9-7 A slurry of 9-6 (150 mg, 0.454 mmol) in was treated with LiOH (38.2 mg, 0.91 mmol) and stirred for 1 hour.
TLC indicated no reaction. LiOH (38.2 mg.) was added and the reaction mixture (still a slurry) was stirred for an additional hour with no reaction. The reaction mixture was concentrated to dryness and dissolved in MeOH and H20. LiOH (114 mg) was added and the reaction mixture was stirred and heated to 750 for 3 hours and stirred at RT for 16 hours resulting in a complete reaction. The reaction mixture is diluted with EtOAc and 10% citric acid. The layers are separated and the organic layer is washed with water and brine, dried, filtered and concentrated to give 9-7 as a white solid.
WO 98/00134 WO 9800134PCT/US97/1 1133 75 IH NMR (400 MHz; CDC13) 8 8.40 IH), 7.95(d, 1H), 7.32 1H), 6.41 1H), 4.84 2H), 4.14-4.11 (in, 2H), 3.96 (in, 2H), 1.50 (s, 9H).
BOON -Q-j OXOO 2 Et 9-
OH
3 Ethyl 1,1 -dimethylethoxycarbonyl)-2,3 [1 indole-8 -yl)carbonylamino)-3 -methvlp~henoxyacetate 9-8 A solution of 9-7 (110 mg, 0.35 mmol) and 10-4 (85.7- mg, 0.35 mmol) in CH2C12 were treated with diisopropylethylamine and PYCLU as described for 23-5 to give 9-8 as a white solid after chromatography in 50% EtOAC/Hexane.
Rf (50% EtOAc/Hexane) 0.34 1 H NMR (400 MHz, CDCI3) 8 8.12 1H), 7.8-7.65 (mn, 2H), 7.59 (s, 1H), 7.36 1H), 6.84 (mn, 2H), 6.39 1H), 4.85 2H), 4.62 2H), 4.29-4.27 2H), 4.15-4.13 2H), 3.96 2H), 2.33 3H), 1.50 (s, 9H), 1.33-1.3 1 3H).
<N 0 HN N 0j\/OC 2 H 9-9
OH
3 204-((2,3 ,4,5 -tetrahydropyrazino-[ 1,2-aindole-8 -yl)carbonylamino)-3 methyiphenoxyacetic acid 9-9 A slurry of 2-8 (0.15 mmnol, 75 ing) and THFIH2O/MeOH was treated with LiOH (13 ing, 0.30 mmol) and stirred for 1 hour. The reaction mixture was diluted with EtOAC and 10% citric acid. The WO 98/00134 PCT/US97/11133 -76layers were separated and the organic layer was washed with H20 and brine. The organic layer was dried, filtered and concentrated to give desired acid as a white solid. This solid is slurried in EtOAc, cooled to -78o and saturated with HC1. The reaction mixture is warmed to 0° and stirred for 15 min. The reaction mixture was concentrated to yield a white solid which was taken up in a EtOAc/hexane/ether mixture and filtered. The solids were washed with ether to give 9-9 as a white solid.
Rf 10/1/1 EtOH/NH40H/H20) 0.16 1 H NMR (400 MHz; D20) 8 8.09 1H), 7.62-7.60 1H), 7.45 (d, 1H), 7.14-7.12 1H), 6.84 1H), 6.79-6.76 (dd, 1H), 6.35 1H), 4.40 2H), 4.07-4.02 4H), 3.23-3.22 2H), 2.15 3H).
r N CO 2
CH
3 9 BOCN 8-Methoxycarbonyl-3-(1,1 -dimethylethoxycarbonyl)- 1, a,2, 3,4.5-hexahvdropyrazino-r 1,2-alindole 9-10 2-6 (0.091 mmol, 30 mg) was dissolved in EtOAc and cooled to 0°C. NaBH3CN (0.45 mmol, 28 mg) was added portion-wise and the reaction was warmed to room temperature for 15 min. The reaction mixture was basified with saturated NaHCO3 and extracted into EtOAc. The organic layer was washed with brine, dried (MgSO4), filtered and concentrated to yield 9-10 as a colorless oil.
Rf (2:1 hexane/ EtOAc)=0.4 1H NMR (400 MHz; CDC13) 8 7.83-7.82 1H), 7.73 1H), 6.40- 6.38 1H), 4.15-4.0 (bs, 2H), 3.85 3H), 3.60-3.56 2H), .305- 2.65 4H), 2.60-2.58 (dd, 1H), 1.50 9H).
WO 98/00134 WO 9800134PCT/US97/1 1133 77 BOCN 1,1-Dimethylethoxycarbonyl)- 1,1a,2,3 ,4,5 -hexahydropvrazino-[ 1.2-alindole-8-carboxylic acid 9-11 9-10 (0.90 mmol, 300 mg) was slurried in THFIH2OIMeOH (2mL/2/2). LiGH (1.8 mmol, 76 mg) was added and the reaction mixture was heated to 50'C. After 0.5 hours, the reaction mixture became homogeneous, and was then stirred at room temperature for an additional 2 hours. The reaction mixture was diluted with 10% citric acid and EtOAc. The layers were separated, and the organic layer was washed with H20 and brine. Drying (MgSO4), filtering and concentrating gave 9-11 as a yellow solid.
Rf (97/3/1 CHCl3IMeOH/HOAC)=0.70 I H NMR (400 MHz; CDCl3) 8 7.90 1H), 7.87 1H), 6.41-6.39 (d, LH), 4.25-4.0 (bs, 2H), 3.65-3.57 (in, 2H), 3.10-3.0 (dd, 1H), 3.02-2.98 d, 1H), 2.98-2.91 (bs, 1H), 2.69-2.66 (bs, IH), 2.62-2.60 (dd, 1H), 1.50 9H).
N 0 (-)BOON N O'CO2Et 9-12
OH
3 I)Ethyl 1,1-Dimethylethoxycarbonyl)- 1,1a,2,3 ,4,5 hexahydro-pyrazino- [1 2 -ajindole-8-yl)carbonylamino)-3 methylp~henoxiyacetate 9-12 A solution of 9-1 1 (250 mg, 0.79 inmol) and 10-4 (193 mng, 0.79 mmol) in CH2Cl2 were treated with diisopropylethylamine and WO 98/00134 PCT/US97/11133 -78 PYCLU as described for 23-5 to give 9-12 as a white solid after chromatography in 50% EtOAC/Hexane.
Rf (50% EtOAc/Hexane) 0.35 IH NMR (400 MHz, CDC13) 5 7.70-7.63 3H), 7.36 1H), 6.82- 6.76 2H), 6.45-6.43 1H), 4.60 2H), 4.30-4.25 2H), 4.21- 4.09 (bs, 2H), 3.59-3.57 2H), 3.10-3.09 1H), 3.05-2.51 3H), 2.78-2.60 1H), 2.29 3H), 1.50 9H), 1.32-1.29 (t,3H).
N O 0 HN N CO 2 H 9-13
CH
3 4-((3-(1,1-Dimethylethoxycarbonyl)- 1,1a,2,3,4,5-hexahydropyrazino-[ 1,2-a]indole-8-yl)carbonylamino)-3-methylphenoxyacetic acid 9-13 A slurry of 9-12 (0.25 mmol, 125 mg) and THF/H20/MeOH was treated with LiOH (21 mg, 0.50 mmol) and heated to 800 for 1 hour. The reaction mixture was diluted with EtOAC and citric acid. The layers were separated and the organic layer was washed with H20 and brine. The organic layer was dried, filtered and concentrated to give desired acid as a white solid. This solid is slurried in EtOAc, cooled to -780 and saturated with HC1. The reaction mixture is warmed to 0° and stirred for 15 min. The reaction mixture was concentrated and the white solid triturated with ether to give 9-13 as a white solid.
Rf 10/1/1 EtOH/NH40H/H20) 0.3 1 H NMR (400 MHz, D20) 5 7.70-7.67 1H), 7.63 1H), 7.11-7.09 1H), 6.86 1H), 6.78-6.76 (dd, IH), 6.69-6.68 1H), 4.57 (s, 2H), 3.93-3.90 (bd, 2H), 3.39-3.31 3H), 3.19-3.07 3H), 2.74- 2.68 (dd, 1H).
WO 98/00134 WO 9800134PCT/US97/11133 79 SCHEME
OH
H
3 0 N
NH
2
OH
Boo 2 O CHCI 3 ref lux
H
3 C N Br COOEt
CS
2 00 3
DMF
NHBoc 10-2 HCI (gas) EtOAc, 000 Li 0 30
*HCI
NH
2
H
3
C'
NHBoc 10-4
COOH
N.
N,
Boo P F 6 10-4 i-Pr 2 NEt, 0H 2 C1 2 WO 98/00134 PCT/US97/11133 80 SCHEME 10 CONTINUED Boc-N N NaOH, H 2 0 MeOH, RT OEt
H
3
C
0 0 Boc-N N- N OH
H
3
C
10-6 HCI (gas) EtOAc, 00C
H
3
C
10-7 1.1 -Dimethylethoxycarbonyl)amino-3-methvlphenol (10-2) To a 1 L round bottomed flask with a stirring bar, reflux condenser and an argon inlet was added 4-amino-3-methylphenol (15.00 g, 121.79 mmol), di-tert-butylpyrocarbonate (27.25 g, 124.84 mmol) and CHC13 (300 mL). This heterogeneous mixture was heated at reflux for 24 h during which time all of the solids dissolved. The mixture was cooled to room temperature and the solid product was collected by filtration. The material was triturated with a mixture of collected on a frit and dried in vacuo to give 21.25 g of 4- WO 98/00134 PCT/US97/11133 -81 (1,1 -dimethylethoxycarbonyl)amino-3-methylphenol 10-2), mp: 143- 144 0
C.
IH NMR (CDC13): 6 1.51 9H), 2.14 3H), 6.08 (br s, 1H), 6.48 2H), 6.60 (br s, 1H), 7.20 j=8.5Hz, 1H).
Ethyl 4-(1,1-Dimethylethoxycarbonyl)amino-3-methylphenoxyacetate (10-3) To a 200 mL round bottomed flask with a stirring bar, and an argon inlet was added 4-(1,1 -dimethylethoxycarbonyl)amino-3methylphenol (5.00 g, 22.39 mmol), Cs2CO3 (14.59 g, 44.78 mmol), DMF (50 mL), and ethyl bromoacetate (2.61 mL, 23.51 mmol). This mixture was stirred vigorously at ambient temperature for 24 h. The mixture was filtered through a frit and the DMF was removed under high vacuum. The residue was dissolved in EtOAc (300 mL) and washed with H20 (2x) and brine Drying (MgSO4), filtration, and removal of the solvent in vacuo gave a solid. This material was triturated with 5% Et20-hexane, the solid was collected by filtration and dried in vacuo to give 5.40 g of ethyl 4-(1,1-dimethylethoxycarbonyl)amino-3-methylphenoxyacetate as a white, crystalline solid.
IH NMR (CDC13): 8 1.29 j=7.2Hz, 3H), 1.51 9H), 2.22 3H), 4.26 j=7.2Hz, 2H), 4.57 2H), 6.08 (br s, 1H), 6.72 2H), 7.56 1H).
Ethyl 4-amino-3-methylphenoxyacetate. hydrochloride (10-4) To a 500 mL round bottomed flask with a gas dispersion tube was added a solution of ethyl 4-(1,1-dimethylethoxycarbonyl)amino-3-methylphenoxyacetate (5.31 g, 17.13 mmol) in EtOAc (200 mL). This solution was cooled in an ice bath and dry HC1 gas was sparged through the solution, vigorously, for 10 min. The resulting mixture was aged for 15 min. at 0°C. The excess HCI gas was removed with a stream of argon and the solvent was removed in vacuo. The product was triturated with 50 mL of EtOAc and collected on a frit.
The crystals were washed with additional EtOAc and dried in vacuo to WO 98/00134 WO 9800134PCTIUS97I1 1133 82 give 4.21 g (100%) of ethyl 4-amino-3-methylphenoxyacetate, hydrochloride as white crystals, mp: 198-200'C.
I H NMR (DMSO-d6): 8 1.21 j=7.lIHz, 3 2.3 3 3H), 4.17 (q, j=7.l1Hz, 2H), 4.78 2H), 6.82 (dd, j=3,9Hz, I1H), 6.92 j=3Hz, I1H), 7.39 j=9Hz, IH), 10.21 (br s, 3H).
Ethyl 1,1-dimethylethoxycarbonyl)piperazin-1I-yl)phenylcarbonylamino)-3-methvlphenoxy)acetate (10-5) To a 100 mL, round bottomed flask with a stirring bar and an argon inlet was added 1, 1 -dimethylethoxycarbonyl) piperazinyl)benzoic acid (0.75 g, 2.45 mmol), ethyl 4-amino-3methylphenoxyacetate hydrochloride (0.60 g, 2.45 mmol), chioro- N,N,N' ,N',-bis (pentamethylene)formamidinium hexafluorophosphate (0.97 g, 2.69 mmol), and CH2CI2 (30 mL). This mixture was cooled in an ice bath and diisopropylethylamine (1.74 mL, 10.0 mmol) was added.
The ice bath was allowed to expire and the solution was stirred at ambient temperature for 48 h. The solution was diluted with CHC13 and washed with 10% aqueous citric acid, saturated aqueous NaHICO3, and brine. Drying (MgSO4), filtration and removal of the solvent in vacuo gave an oil. This material was chromatographed on 75 g of silica gel using 50% EtOAc-hexane as eluant. There was obtained 1.22 g (100%) of ethyl 1,1-dimethylethoxycarbonyl)piperazin- 1yl)phenylcarbonylamino)-3 -methylphenoxy)acetate as a crystalline solid.
1 H NMR (CDCl3): 5 1.30 j=7.1IHz, 3H), 1.49 9H), 2.27 3H), 3.27 (in, 4H1), 3.59 (mn, 4H1), 4.26 j=7.lHz, 2H), 4.59 211), 6.75 (mn, 2H1), 6.90 j=8.8Hz, 2H), 7.54 (br s, I 7.65 (mn, ILH), 7.79 (d, j=8.8Hz, 2H1).
1, 1 -Dimethylethoxycarbonyl)piperazin- I -yl)phenylcarbonylamino)-3 -methylphenoxy)acetic acid (10-6) To a 100 ML. round bottomed flask with a stirring bar and an argon inlet was added ethyl 1,1 -dimethylethoxycarbonyl)piperazin- I -yl)phenylcarbonylamino)-3 -inethylphenoxy)acetate (1.22 g, 2.45 mmol) and 20 mL. of CH3 OH. To this solution was WO 98/00134 PCT/US97/11133 83 added aqueous NaOH (10 mL of a IN solution). The mixture was stirred at ambient temperature for 18 h. The mixture was neutralized with 10 mL of IN HCI and diluted with H20. The product was collected on a frit and washed with a little H20. This material was dried in vacuo to give 0.953 g of 2-(4-(4-(4-(1,1-dimethylethoxycarbonyl)piperazin-1 -yl)phenylcarbonylamino)- 3 -methylphenoxy)acetic acid as a white solid.
1 H NMR (CDC13): 8 1.49 9H), 2.26 3H), 3.27 4H), 3.58 (m, 4H), 4.57 2H), 6.74 2H), 6.90 j=8.5 Hz, 2H), 7.60 1H), 7.65 IH), 7.79 j=8.5 Hz, 2H).
2-(4-(4-(1-Piperazinyl)phenylcarbonylamino)- 3 -methylphenoxy)acetic acid. dihydrochloride (10-7) To a 200 mL round bottomed flask equipped with a stirring bar and a gas dispersion tube was added 2-(4-(4-(4-(1,1-dimethylethoxycarbonyl)piperazin- 1 -yl)phenylcarbonylamino)- 3 -methylphenoxy)acetic acid (0.95 g, 2.03 mmol) and 100 mL of dry EtOAc.
This well stirred suspension was cooled in an ice bath and HCI gas was sparged through the solution for 15 min. This mixture was aged min. at 0°C then the excess HCI was removed with a stream of argon and the EtOAc was removed in vacuo. The product was triturated with EtOAc, collected on a frit and dried in vacuo to give 895 mg of (1-piperazinyl)phenylcarbonylamino)- 3 -methylphenoxy)acetic acid, dihydrochloride, mp: >250 0
C.
1 H NMR (DMSO-d6): 8 2.16 3H), 3.20 4H), 3.54 4H), 4.66 2H), 6.72 j=3Hz, 1H), 6.75 1H), 7.06 j=9.0Hz, 2H), 7.15 j=3.0Hz, IH), 7.90 j=9.0Hz, 2H), 9.43 (br s, 1H), 9.56 1H).
WO 98/00134 WO 9800134PCTIUS97/11133 84 SCHEME 11
'OH
11-1 R.A. Glennon.; et. al.
J. Med. Chem. 1992 35 734.
OH
SOCI
2 MeOH 00 H 3 11-2
'OH
KI
K
2 00 3 Acetone CK C0 2 Et 00CH 3 ICO 2 Et NaH Toluene Na+ 11-4 WO 98/00134 WO 98/0 134PCTIUS97/11133 85 SCHEME 11I (CONT'D' NaOH EtOH reflux 0 Br0 11-5 (EtO) 2 P(0)CHCN- Nal
CN
11-6
B
2
H
6
THE
,HCI
11-7.
NaOH HCOEt \-4x H H 11-8 WO 98/00134 WO 9800134PCTIUS97/11133 86 I I (C()NjlrT-ll 11-8 1j-- I1. NaBH 4 2. B0C 2 0 TEA OtBu B r Pd(OAc)2 1,3 bis diphenyl phosphino propane TEA DMSO /MeOH 11-10 11-11 OtBu o z "N, OtBu
OH
WO 98/00134 WO 9800134PCTIUS97/11 133 87 SCHEME -11 (CONTD) 1. oxalyl chloride DMF cat.
2.
H
2 N P \I \C 2 Et
HCI
1- 3 C 1 11-13 o CO 2 Et 1. LiOH 2. HCI 11-14 WO 98/00134 PCT/US97/11133 -88 0 O Br NHOCH3 11-2 Br OH Methyl 4-bromo-2-hydroxy benzoate 11-2 200 mL of MeOH was cooled to 0°C and treated dropwise with thionyl chloride (30 mL, 0.4 mole) so that the reaction temperature was kept below 15 0 C throughout the addition. A solution of 11-1 (12.7g, prepared by the method of R. Glannon et al., J. Med. Chem.
1992 35, 734, 0.0585 mol) in 50 mL MeOH was added to the reaction and the mixture was warmed to room temperature and stirred for 48 hours, then heated to 60 0 C for 7 hours. The volatile components were removed under high vacuum and the residue chromatographed (silica, gradient straight hexanes to 10% EtOAc/Hexanes) to give 11-2 as a bright yellow oil that solidified on standing.
Rf EtOAc/Hexanes) 0.6 1 H NMR (400 MHz, CDC13) 8 10.8 1H), 7.68 1H), 7.19 1H), 7.03 1H), 3.95 3H).
0
OCH
3 11-3 Br 0 'O CO 2 Et Methyl 4-Bromo-2-(ethyl acet-2-vloxv)-benzoate 11-3 A solution of 11-2 (26.6 g, 0.115 mole) in acetone (80 mL) was treated with chloroethylacetate (14 g, 0.115 mole), potassium iodide (3 g, 0.018 mole), and potassium carbonate (31.7 g, 0.23 mole) and heated to 50 0 C with vigorous stirring for 2 hours. The suspension was filtered, evaporated to dryness, resuspended in ether, re-filtered and concentrated to give 11-3 as a golden-brown oil.
WO 98/00134 PCT/US97/11133 -89- Rf (10% EtOAc/Hexanes) 0.11 1H NMR (400 MHz, CDC13) 8 7.7 1H), 7.2 1H), 7.04 1H), 4.7 2H), 4.3 2H), 3.9 3H), 1.3 3H).
0 BrO 6-Bromo-benzofuran-3-one 11-5 Sodium hydride (5.52 g, 60% dispersion in oil, 0.138 mole) was placed in a 3L vessel equipped with an overhead stirrer, argon line, reflux condenser and addition funnel. Toluene (215 mL) was added and the solution heated to reflux. A solution of 11-3 (36.3 g, 0.115 mole) in 100 mL toluene was added dropwise over 1.5 hours to give an orange suspension. The reaction was heated for an additional 3 hours, then cooled to room temperature and filtered through a glass-fiber filter.
The cake was washed with 500 mL Et20 and dried under vacuum to give 11-4 as a sandy orange solid (streaks from baseline in 30% EtOAc/ Hexanes). A suspension of 11-4 (5 g, 0.016 mol) in EtOH (20 mL) was added to a solution of NaOH (4 g, 0.1 mole) in H20 (35 mL). The reaction was heated to reflux for 1.25 hours, then cooled and the volatile components removed under vacuum. The residue was suspended in 200 mL 6N HC1 and 200 mL CHC13 and stirred vigorously for 0.5 hours. The layers were separated and the aqueous layer was washed with CHC13. The organic layers were combined, dried with MgS04, filtered and evaporated to give 11-5 as a deep red solid.
Rf (20% EtOAc/Hexanes) 0.61 1H NMR (400 MHz, CDC13) 8 7.53 IH), 7.35 1H), 7.24 (d, 1H), 4.64 2H).
WO 98/00134 PCT/US97/11133
CN
Br
O
(6-Bromo-benzofuran-3-yl)-acetonitrile 11-6 Sodium hydride (0.7 g, 0.0173 mol) was washed with hexanes then suspended in THF (8 mL). Diethyl cyanomethylphosphonate (3.07 g, 0.0173 mol) was added dropwise and stirred 10 minutes to give clear yellow solution. A solution of 11-5 (3.38 g, 0.0158 mol) in THF (20 mL) was added dropwise, the solution stirred for 0.5 hour, then heated to reflux for 0.5 hour. A solution of 6N HCL (3.4 mL) was added and the THF was removed under vacuum.
The concentrated solution was diluted with Et20 and washed with 4 X mL 6N HCI, the ether layer was dried (MgSO4), filtered and evaporated. The residue was chromatographed (20% EtOAc/Hexanes) to give 11-6 as an orange solid.
Rf (20% EtOAc/Hexanes) 0.45 1 H NMR (400 MHz, CDC13) 8 7.7 1H), 7.65 1H), 7.45 2H), 3.75 2H).
NH
2
HCI
11-7 BrJ O 2-(6-Bromo-benzofuran-3-vl)-ethylamine 11-7 A solution of 11-6 (10.9 g, 0.0462 mol) in THF (100 mL) was cooled to 0°C and treated dropwise with BH3/THF solution (1Molar, 110.8 mL, 0.11 mol). The cold bath was removed and the reaction stirred overnight. MeOH was carefully added (20 mL) and the solvent removed under vacuum.. The residue was dissolved in 50 mL WO 98/00134 PCT/US97/11133 -91 MeOH and 400 mL MeOH saturated with HCI and heated to reflux for 3 hours. The volatile components were removed under vacuum and the residue was suspended in Et20. A bright yellow solid percipitated and was collected and washed five times with Et20, then dried under vacuum to give 11-7 as a tan solid.
Rf MeOH/ CHC13 saturated with NH3) 0.23 1H NMR (400 MHz, CD30D) 5 7.72 2H), 7.6 1H), 7.45 (d, 1H), 3.25 2H), 3.06 2H).
~OBu 11-10 Br O 7-bromo-2-(1,1 -dimethylethoxycarbonyl)- 1,2,3,4-tetrahydro-9Hpyridof3,4-blbenzofuran 11-10 A solution of 11-7 (11.8 g, 0.043 mol) in MeOH (150 mL) was diluted with CHC13 (400 mL) and washed with 75 mL 1N NaOH.
The aqueous layer was backwashed with CHC13, the organic layers were combined, evaporated, and the residue azeotroped with toluene to remove residual water. The resulting brown oil was treated with 200 mL ethyl formate and brought to reflux for 1.5 hours until the solution was homogenous. The solvent was removed, the residue was dissolved in 200 mL CHC13, washed with 30 mL IN HC1, brine, dried over MgS04, filtered and evaporated to give 11-8 as a solid that was used without further purification. Rf MeOH/ CHC13 saturated with NH3) 0.47 IH NMR (400 MHz, CD30D) 8 8.04 1H), 7.67 1H), 7.63 (s, 1H), 7.55 1H), 7.4 1H), 3.53 2H), 2.4 2H). Solid 11-8 was crushed and placed in a flask equipped with an overhead stirrer.
Polyphosphoric acid (20 mL) and CHC13 (5 mL) was added and the reaction heated to 80 0 C for 1.5 hours. The reaction was removed from the oil bath and 250 mL H20, 30 mL NH40H, and 200 mL CHC13 was added and stirred vigorously. The resulting opaque mixture was WO 98/00134 PCT/US97/11133 -92transferred to a separatory funnel, the layers separated, and the aqueous layer was extracted repeatedly with CHC13. The organic layers were combined, filtered through a pad of SolkaFloc, dried over Na2SO4, filtered and evaporated to give the intermediate cyclic imine as a pale yellow foam. A 5g sample of this material (0.02 mol) was dissolved in 200 mL MeOH and 20 mL IN HC1. Sodium borohydride 0.04 mol) was added in portions. After 0.5 hours the solvent was removed and the residue was partitioned between saturated NaHCO3 and CHC13. The aqueous layer was extracted with CHC13, the organic layers were combined, dried (Na2SO3) and filtered to give a clear yellow solution. (Rf amine MeOH/ CHC13 saturated with NH3) 0.45). Di-tert-butyl dicarbonate (4.5 g, 0.02 mol) and TEA (2.8 mL, 0.02 mol) were added and the solution stirred overnight, then washed with 10% KHSO4. The organic layer was evaporated and the residue was chromatographed in 10% EtOAc/Hexanes to give 11-10 as a white solid.
Rf (20% EtOAc/Hexanes) 0.60 1 H NMR (400 MHz, CDC13) 5 7.6 1H), 7.35 1H), 7.3 1H), 4.56 (bs, 2H), 3.75 (bs, 2H), 2.7 (bs, 2H), 1.5 9H).
N I OCH 3 11-11 OtBu Methyl 2-(1,1-dimethylethoxycarbonyl)-1,2,3,4-tetrahydro-9Hpyridof3,4-b1benzofuran-7-vl carboxvlate 11-11 A solution of 11-10 (lg, 2.84 mmol), 1,3 di-phenyl phosphino propane (0.35 g, 0.852 mmol) in DMSO/MeOH 15 mL) was treated with TEA (1.5 mL, 10.7 mmol) and thouroughly purged with carbon monoxide, then treated with Pd(OAc)2 (0.191g, 0.852 mmol) and warmed to 80 0 C. After heating overnight, the reaction was cooled, treated with an additional amount of 1,3 di-phenyl phosphino WO 98/00134 PCT/US97/11133 -93 propane (0.35g, 0.852 mmol), re-purged with carbon monoxide and treated with an additional amount of Pd(OAc)2 (0.191 g, 0.852 mmol).
After heating for three days the reaction was evaporated, partitioned between water and CHC13, and the water layer washed with CHC13.
The organic layers were combined, dried over MgSO4, filtered through a sintered glass filter and concentrated. The residue was chromatographed (silica, gradient 5% EtOAc/Hexanes to EtOAc/Hexanes) to give 11-11 as a yellow oil.
Rf (20% EtOAc/Hexanes) IH NMR (400 MHz, CDC13) 8 8.13 1H), 7.96 1H), 7.48 1H), 4.6 (bs, 2H), 3.95 3H), 3.75 (bs, 2H), 2.75 (bs, 2H), 1.5 9H).
SOH 11-12 'N 1
OH
OtBu 2-(1,1 -dimethylethoxycarbonyl)-1,2,3,4-tetrahydro-9H-pyrido[3,4blbenzofuran-7-yl carboxylic acid 11-12 A solution of 11-11 (0.79 g, 2.38 mmol) in 1:1 THF/MeOH mL) was treated with LiOH (0.95 g, 23.8 mmol) in 10 mL of and heated to 30 0 C overnight. The solvents were removed under vacuum and the residue was dissolved in H20 and acidified with KHSO4 and extracted with EtOAc. The organic layer was dried over MgSO4, filtered and evaporated to give 11-12 as an off-white solid.
Rf (97:3:1 CHCl3/MeOH/HOAc) 0.27 1H NMR (400 MHz, CDC13) 6 8.2 1H), 8.0 1H), 7.50 1H), 4.65 (bs, 2H), 3.8 (bs, 2H), 2.75 (bs, 2H), 1.52 9H).
WO 98/00134 PCT/US97/11133 -94- N ,~ON
OO
2 Et 11-13 OtBu H 3
C
Ethyl 4-(2-(1,1-dimethylethoxycarbonyl)-1,2,3,4-tetrahydro-9Hpyrido[3,4-b]benzofuran-7-yl)carbonylamino)-3-methylphenoxyacetate 11-13 A solution of 11-12 (0.1 g, 0.321 mmol) in CH2C12 mL) was treated with oxalyl chloride (0.06 mL, 0.7 mmol) and two drops of DMF. After stirring at room temperature for 0.5 hours, the reaction was diluted with benzene and evaporated. The resulting acid chloride was dissolved in CHC13 (5 mL) and treated with 10-4 (0.075 g, 0.26 mmol), then cooled to 0°C and treated with diisopropylethylamine (0.167 mL, 0.3 mmol). The reaction was warmed to room temperature and stirred overnight, then evaporated, partitioned between EtOAc and KHSO4, and the EtOAc layer was washed again with 10% KHSO4.
The EtOAc layer was washed with brine, dried over MgSO4, filtered and evaporated to give 11-13.
Rf (60% EtOAc/Hexanes) 0.64 1H NMR (400 MHz, CDC13) 8 8.0 1H), 7.73 2H), 7.62 1H), 7.53 1H), 6.85 1H), 6.8 1H), 4.62 2H), 4.61 2H), 4.29 2H), 3.68 (bs, 2H), 2.75 (bs, 2H), 2.30 3H), 1.5 9H), 1.3 (t, 3H).
WO 98/00134 PCT/US97/11133 HN HN 0 CO 2 H 11-14 HN O
HONH
H
3
C
-dimethylethoxycarbonyl)- 1,2,3,4-tetrahydro-9H-pyrido[3,4blbenzofuran-7-yl)carbonylamino)-3-methylphenoxy acetic acid 11-14 A solution of 11-13 (0.145 g, 0.285 mmol) in 1:1 THF/MeOH (6 mL) was treated with LiOH (0.114 g, 28.5 mmol) dissolved in 6 mL of H20. After stirring for 1 hour at room temperature the solvents were removed, the residue dissolved in H20/EtOAc/10% KHSO4 and the layers separted. The water layer was washed with EtOAc, the organic layers were combined, dried, and filtered to give the intermediate acid as a yellow oil. This material was dissolved in EtOAc (10 mL), cooled to -78 0 C, the solution saturated with HCI gas and warmed to 0°C for 0.5 hours, then concentrated and dried under vacuum to give 11-14 as fluffy solid.
Rf (9:1:1 EtOH/H20/NH40H) 0.78 1H NMR (400 MHz, D20) 6 7.84 1H), 7.65 1H), 7.5 1H), 7.1 1H), 6.81 1H), 6.72 1H), 4.28 2H), 3.8 (bs, 2H), 2.96 2H), 2.6 (bs, 2H), 2.13 3H).
WO 98/00134 WO 980134P1US97I11 133 96 SCHEME 12
OH
H
3 C 1
HO
3 S /N 2
+CI
OH
H
3
C
NH
2 2. Na 2
S
2
O
4 12-1 12-2 Boc 2 O CHCI 3 ref lux
OH
H
3
C
NHBoc Br 11 COOEt
CS
2 00 3
DMF
12-3
H
3
C.
HOI (gas) EtOAc, 0 0
C
NHBoo 12-4 WO 98/00134 PTU9/13 PCTIUS97/11133 97 SCHEME 12 CONTINUED
H
3
C
COOEt
.HCI
Boc-N N\/COOH PI PF 6 7
NH
2 i-Pr 2 NEt, CH 2 01 2 COOEt LiOH DME H 2 0 12-6 12-7 HCI (gas) EtOAc, 000
.HCI
0 OH 3
HN\_-N-O-COOH
12-8 WO 98/00134 PCT/US97/11133 -98 2-Methvl-3-aminophenol (12-2) To a 500 mL erlenmeyer flask was added sulfanilic acid (14.38 g, 83 mmol), sodium carbonate (4.29 g, 40.5 mmol) and distilled (83 mL), when all of the sulfanilic acid dissolved, the solution was cooled in an ice bath to 0°C and a solution of sodium nitrite (6.16 g, 89.3 mmol) in H20 (15 mL) was added in one portion. The ice bath was allowed to warm to +15°C and the mixture was stirred at this temperature 1 h. The reaction mixture was poured onto a mixture of ice (100 g) and 12N HCI (17.4 mL). The ice was allowed to melt and the solid diazonium salt was collected by suction filtration on a scintered glass frit. 0-cresol (9.10 g, 84.1 mmol) was dissolved in a solution of NaOH (1.21 g, 30.25 mmol) in H20 (100 mL). This solution was cooled to 0°C and the solid diazonium salt was added in one portion.
This well stirred mixture was maintained at +15°C for 4 h. The temperature of the mixture was raised to +60 0 C and sodium dithionite g, 1.03 mol) was added portionwise. The reaction was allowed to proceed at +60 0 C for 15 min. The mixture was cooled to room temperature, diluted with saturated aqueous NaHCO3 and extracted with EtOAc. The EtOAc extract was dried (MgSO4), filtered, concentrated in vacuo and chromatographed on 100 g of silica gel using 1:3 EtOAchexane as eluant. There was obtained 2-methyl-3-amino-phenol as a tan solid.
1 H NMR (CDC13) 8 2.18 3H), 3.35 (br s, 2H), 4.24 (br s, 1H), 6.42 1H), 6.50 1H), 6.61 j=9Hz, 1H).
4- 1,1 -Dimethvlethoxycarbonvl)amino-2-methylphenol (12-4) Using a method similar to that described for compound 10-2, 4-(1,1-dimethylethoxycarbonyl)amino-2-methylphenol was prepared.
1 H NMR (CDC13): 8 1.51 9H), 2.21 3H), 4.62 (br s, 1H), 6.22 (br s, 1H), 6.62 1H), 6.98 1H), 7.08 (br s, 1H).
WO 98/00134 WO 9800134PCTIUS97/1 1133 -99- Ethyl 1,1-dimethylethoxycarbonyl)amino-2-methylphenoxyacetate (12-4) Using a method similar to that described for compound 103 ethyl 4-(1.1 -dimethylethoxycarbonyl)amino-2-methylphenoxyacetate was prepared.
I H NMR (CDCl3): 5 1.29 j=7.2Hz, 3H1), 1.52 9H), 2.28 3H), 4.25 j=7.2Hz, 2H), 4.58 2H), 6.25 (br s, 1H), 6.65 1 7.08 (br d, j=8.5Hz, I1H), 7.17 (br s, I1H).
Ethyl 4-amino-2-methylphenoxyacetate. hydrochloride (12-5) Using a method similar to that described for compound 10-4, ethyl 4-amino-2-methylphenoxyacetate, hydrochloride was prepared.
I H NMR (CD3OD): 8 1.21 j=7.lIHz, 3H), 2.35 311), 4.25 (q, j=7.l1Hz, 2H), 4.81 2H), 6.95 j=,9Hz, I 7.21 (in, 2H1).
Ethyl 1,1 -dimethylethoxycarbonyl)piperazn- 1 -yl)phenylcarbonylamino)-2-inethvlphenoxy)acetate (12-6), Using a method similar to that described for compound 10-5, ethyl 1,1 -dimethylethoxycarbonyl)piperazin- 1yl)phenylcarbonylamino)-2-methylphenoxy)acetate was prepared.
I
1 H NMR (DMSO-d6): 8 1.41 j=7Hz, 3H1), 1.42 9H), 2.20 3H), 2.50 (in, 411), 3.29 (mn, 411), 4.29 j=7Hz, 2H), 4.78 2H), 6.80 (d, Hz, 1H1), 7.01 j=8.8Hz, 2H1), 7.50 (mn, 2H1), 7.86 j=8.8Hz, 2H), 9.68 ILH).
1,1 -dimethylethoxycarbonyl)piperazin- 1 -yl)phenylcarbonylanmino)-2-methylphenoxy)acetic acid (12-7) Using a method similar to that described for compound 10-6, 1,1-dirnethylethoxycarbonyl)piperazin-1I-yl)phenylcarbonylamino)-2-methylphenoxy)acetic acid was prepared and used in the next step without purification.
WO 98/00134 PCT/US97/11 133 -100- -Piperazinyl)phenylcarbonylamino)-2-methylphenoxy)acetic acid, dihydrochloride (12-9) Using a -method similar to that described for compound I1-, 1-piperazinyl)phenylcarbonylamino)-2-methyl phenoxy)acetic acid, dihydrochioride was prepared, mp: >250TC.
I H NMR (D20): 8 2.15 3H), 3.29 (in, 4H), 3.48 (in, 4H), 4.65 (s, 2H), 6.76 j=8.5Hz, 1H), 7.04 j=8.6, 7.17 (in, 2H), 7.69 (d, j=8.6Hz, 2H).
WO 98/00134 WO 9800134PCTIUS97/1 1133 -101- SCHEME 13
COGH
H
3
C
NH
2 MeOH HOI RT
COOCH
3
H
3
CN
NH
2 13-1 13-2
(CICH
2
CH
2 2
NH*HCI
n-BuOH, ref lux
H
3 0 HN N COO(CH 3 n-Bu)
H
3 0 Bo 2 OCHI BocN N N COO(CH 3 n-Bu) 13-4 1N NaOH EtOH, reflux
H
3
C
BocN NIL COOH 13-5 WO 98/00134 WO 9800134PCTIUS97/1 1133 -102- SCHEME 13 CONTINUED
OH
NH
2 13-6 Bo2L9 CHC1 3 ref lux
OH
'-BrCH 2 000Et
-~CS
2
CO
3
DMF
NHBoc 13-7 HOI (g) EtOAc, 000
'HCI
NH
2 13-8 WO 98/00134 PCT/US97/11133 -103
*PF
6 No 13-5 13-9 i-Pr 2 NEt, CH 2 C12
H
3
C
BocN N 13-10 /-COOEt N
H&
LiOH
HCI
H
3
C
HN N N- C0 2
H
H/ H Methyl 3-methyl-4-aminobenzoate (13-2) To a 500 mL round bottomed flask equipped with a stirring bar, reflux condenser and a drying tube was added 3-methyl-4aminobenzoic acid (8.00 g, 52.92 mmol) and anhydrous methanol (300 mL). This solution was saturated with anhydrous HCI gas and the mixture was stirred at ambient temperature for 26 h. The methanol and HCI were removed in vacuo and the solid product was suspended in 400 mL of EtOAc. This mixture was made basic by careful addition of aqueous NaHCO3 solution. The layers were separated and the organic phase was washed with brine, dried (MgSO4), filtered and concentrated in vacuo to give methyl 3-methyl-4-aminobenzoate as a yellow solid.
1 H NMR (CDC13): 8 2.18 3H), 3.85 3H), 3.99 (br s, 2H), 6.63 j=8.3 Hz, IH), 7.75 3H).
WO 98/00134 PCT/US97/11133 104- Methyl and n-Butvl 3-methyl-4-(l-piperazinvl)benzoate (13-3) To a IL round bottomed flask with a stirring bar, reflux condenser, and an argon inlet was added methyl 3-methyl-4aminobenzoate (8.74 g, 52.74 mmol), bis 2-chloroethylamine hydrochloride (9.81 g, 55.0 mmol) and n-BuOH (175 mL). This mixture was heated at reflux for 8 days. The n-BuOH was removed in vacuo and the residue was partitioned between EtOAc (200 mL) and 0.2N HCI (300 mL). The layers were separated and the organic phase was extracted with another 100 mL portion of 0.2 N HC1. The aqueous phases were combined and made basic with solid NaHCO3. This mixture was extracted with EtOAc (2 x 200 mL) and the combined EtOAc extracts were washed with brine, dried (Na2SO4), filtered and concentrated in vacuo to give 10.39 g of a mixture of ethyl and n-butyl 3-methyl-4-(1-piperazinyl)benzoates. This material was used in the next step without further purification.
Methyl and n-Butyl 3-methyl-4-(1-(4-(1,1-dimethylethoxy)carbonyl)piperazinyl)benzoates (13-4) To a 1L round bottomed flask equipped with a stirring bar and an argon inlet was added a mixture of ethyl and n-butyl 3-methyl-4- (1-piperazinyl)benzoates (10.39 g, 44.35 mmol based on the mw of the methyl ester only; vide supra), di-tert-butyldicarbonate (14.67 g, 67.22 mmol) and dry CHC13 (100 mL). This solution was heated at reflux for 19 h. The cooled reaction mixture was concentrated in vacuo and the residue was chromatographed on silica gel (300 g) using 40% EtOAchexane as eluant to give 5.0 g of a mixture of methyl and n-butyl 3methyl-4-(l-(4-(1,1-dimethylethoxy)carbonyl)piperazinyl)benzoates.
This material was used in the next step without further purification.
3-Methyl-4-(1-(4-(1,1 -dimethylethoxy)carbonyl)piperazinyl)benzoic acid (13-5) The product from the preceding step, a mixture of methyl and n-butyl 3-methyl-4-(1-(4-(1,1-dimethylethoxy)carbonyl)piperazinyl)benzoates (5.00 g) was dissolved in 95% EtOH (100 mL) and WO 98/00134 PCT/US97/11133 105- NaOH (10 g, 250 mmol) was added. This solution was heated at reflux for 2 h. The mixture was cooled to room temperature and the EtOH was removed in vacuo. The residue was dissolved in 300 mL of This solution was washed with Et20 (200 mL) then acidified with aqueous citric acid. This mixture was extracted with EtOAc (2 x 250 mL). The combined EtOAc extracts were washed with H20 and brine.
Drying (MgSO4), filtration and removal of the solvent in vacuo gave an oil. This material was chromatographed on 75 g of silica gel using EtOAc-hexane as eluant. The chromatographed product was recrystallized from 15 mL of 20% EtOAc-hexane to give 228 mg of 3methyl-4-(1-(4-(1,1-dimethylethoxy)carbonyl)piperazinyl)benzoic acid as white crystals.
1 H NMR (CDC13): 8 1.49 9H), 2.35 3H), 2.94 4H), 3.59 (m, 4H), 6.99 j=8Hz, 1H), 7.92 2H), 11.65 (br s, 1H).
4-(1,1-Dimethylethoxvcarbonvlamino)phenol (13-7) To a 500 mL round bottomed flask with a stirring bar, reflux condenser and an argon inlet was added 4-aminophenol (10.00 g, 91.63 mmol), di-tert-butyldicarbonate (20.00 g, 91.63 mmol) and dry CHC13 (250 mL). This solution was heated at reflux for 6 h. The mixture was cooled in an ice bath and the product was collected by filtration. The product was washed with a little cold CHC13 and dried in vacuo to give 16.43 of 4-(1,1-dimethylethoxycarbonylamino)phenol as white crystals, mp: 142-143 0
C.
1H NMR (CDC13): 6 1.51 9H), 5.27 (br s, 1H), 6.34 (br s, 1H), 6.72 j=8Hz, 2H), 7.16 j=8Hz, 2H).
Ethyl 11 -dimethylethoxycarbonylaminophenoxy)acetate (13-8) To a 500 mL round bottomed flask with a stirring bar and an argon inlet was added 4-(1,1-dimethylethoxycarbonylamino)phenol (8.20 g, 39.19 mmol), Cs2CO3 (25.54 g, 78.38 mmol), DMF (75 mL), and ethyl bromoacetate (4.78 mL, 43.11 mmol). This heterogeneous mixture was stirred at ambient temperature for 3.5 h. The mixture was diluted with a little CHC13 and filtered through a frit to remove the WO 98/00134 PCT/US97/11133 106 salts. The DMF was removed under high vacuum and the residue was suspended in 500 mL of EtOAc. This mixture was washed with and brine. Drying (MgSO4), filtration and removal of the solvent in vacuo, gave an oil. This material was chromatographed on 400g of silica gel using 20% EtOAc-hexane as eluant. There was obtained 11.9 g (100%) of ethyl (4-(1,1-dimethylethoxycarbonylamino)phenoxy)acetate as an oil.
1 H NMR (CDC13): 6 1.29 j=6.4Hz, 3H), 1.50 9H), 4.25 (q, j=6.4Hz, 2H), 4.57 2H), 6.36 (br s 1H), 6.84 j=8Hz, 2H), 7.26 j=8Hz, 2H).
Ethyl 4-aminophenoxvacetate, hydrochloride (13-9) To a 500 mL round bottomed flask with a stirring bar and a gas dispersion tube was added ethyl (4-(1,1-dimethylethoxycarbonylamino)phenoxy)acetate (11.9 g, 40.29 mmol) and dry EtOAc. This solution was cooled in an ice bath and saturated with anhydrous HCI gas over 15 min. The resulting suspension was aged 30 min. at 0°C. The excess HCI was removed with a stream of argon and the EtOAc was removed in vacuo. The solid product was triturated with EtOAc, collected on a frit, and dried in vacuo at room temperature to give 7.33 g of ethyl 4-aminophenoxyacetate, hydrochloride as a white crystalline soild.
1 H NMR (DMSO-d6): 8 1.211 j=6.4Hz, 3H), 4.16 j=6.4Hz, 2H), 4.82 2H), 7.05 j=8Hz, 2H), 7.34 j=8Hz, 2H), 10.27 (br s, 3H).
Ethyl 2-(4-(4-(3-methyl-4-(1,1 -dimethylethoxycarbonyl)piperazin- 1v)phenylcarbonvlamino)phenoxy)acetate (13-10) In a manner similar to that described for compound 10-5, using acid 13-5 and aniline 13-8, ethyl 2-(4-(4-(3-methyl-4-(1,1dimethylethoxycarbonyl)piperazin-1 -yl)phenyl-carbonylamino)phenoxy)acetate was prepared.
1 H NMR (CDC13): 8 1.29 j=7.0Hz, 3H), 1.48 9H), 2.37 3H), 2.95 4H), 3.59 4H), 4.25 j= 7.Hz, 2H), 4.61 2H), 6.95 (d, 2H), 7.03 j=8.5Hz, 1H), 7.55 j=8.5Hz, 2H), 7.62 (m, 2H).
WO 98/00134 PCT/US97/11133 107 2-(4-(4-(3-Methyl-4-(1-piperazinyl)phenylcarbonylamino)phenoxy)acetic acid (13-11) To a 100 mL round bottomed flask with a stirring bar was added ethyl 2-(4-(4-(3-methyl-4-(1,1-dimethylethoxycarbonyl)piperazin-l-yl)phenylcarbonylamino)phenoxy)acetate (311 mg, 0.63 mmol), MeOH (5 mL), THF (20 mL), and IN LiOH (20 mL). This solution was stirred at ambient temperature for 24 h. The solvents were removed in vacuo and the residue was acidified with 10% aqueous citric acid. This mixture was extracted with EtOAc. The EtOAc extract was dried (MgSO4), filitered and cooled to 0°C in an ice bath. This solution was saturated with dry HCI gas and aged 30 min. at 0°C. The excess HCI was removed with a stream of argon and the solvent was removed in vacuo. The product was purified by preparative reverse phase HPLC using a H20-CH3CN gradient on a Waters C-18 column. Obtained 105 mg of 2-(4-(4-(3-methyl-4-( 1 -piperazinyl)phenylcarbonylamino)phenoxy)acetic acid as a white solid.
1 H NMR (D20): 6 2.28 3H), 3.17 4H), 3.35 4H), 4.55 (s, 2H), 6.94 j=7.5Hz, 2H), 7.12 j=8.2Hz, 1H), 7.33 2H), 7.63 2H).
WO 98/00134 WO 9800134PCT/US97/11133 108 SCHEME 14 HN 0
BOC
2 O THF ref lux Boc-N N-&NO 2 14-1 Pd-C
H
2 (45 psig) EtOAc-MeGH Boc-N-/N NH 2 HO\/aCOOH Cl *PF 6 14-3 i-Pr 2 NEt, CH 2
CI
2
H
Boc-N N N-
\/OH
0 BrCH 2 000Bu t
CS
2 00 3 DMF, RT 14-4
H
N-COOBut 145 0 HOI (g) EtOAc, 0 0
C
H NN N 1
COOH
14-6 WO 98/00134 PTU9/13 PCTIUS97/11133 109 1 -Nitrop~henyl 1.1-dimethvlethoxycarbonvl)piperazine (14-2) To a 200 mL round bottomed flask was added 1 -nitrophenylpiperazine (5.01 g, 24.2 inmol), di-tert-butyldicarbonate (5.85 g, 26.8 mimol), triethylamnine (3.87 mL, 27.8 mmol), and dry THF (100 mL). This solution was heated at reflux for 18 h. The mixture was cooled to room temperature and the solvents were removed in vacuo.
The residue was dissolved in EtOAc and washed with 10% aqueous citric acid, H20, and brine. Drying (MgSO4), filtration, and removal of the solvent in vacuc gave 7.7 g of 1-nitrophenyl-4-(1,1I-dimethylethoxycarbonyl)piperazine as a yellow solid.
1 H NMR (CDCl3): 8 1.42 9H), 3.41 (in, 4H), 3.62 (in, 4H), 6.81 (d, 2H), 8.16 j=-8.5Hz, 2H).
1 .1-Dimethylethoxvcarbonvl)piperazin- 1 -l)aniline (14-3) To a 250 mL Parr haydrogenation bottle was added 1 nitrophenyl-4-( 1,1-dimethylethoxycarbonyl)piperazine (4.5 g, 16.34 inmol), EtOAc (81 mL), MeOH (27 mL), and 5% Pd-C (0.18 This mixture was hydrogenated under 45 psig H2 at ambient temperature for 24 h. The mixture was filtered through a celite pad and the solvents were removed in vacuo. The residue was chromatographed on 250 g of silica gel using 70% EtOAc-hexane as eluant. There was obtained 3.43 g of 1,1 -dimethylethoxycarbonyl)piperazin-1I-yl)aniline as a yellow solid.
I H NMR (CDCl3): 8 1.49 9H), 2.98 (mn, 4H), 3.49 (br s, 2H), 3.57 (in, 4H), 6.66 j=9Hz, 2H), 6.82 j=9Hz, 2H).
,1 -dimethylethoxycarbonyl)piperazin- 1 -yl)phenyl)-4hydroxybenzamide (14-4) To a 100 ml- round bottomed flask with a stirring bar and an argon inlet was added 1,1 -dirnethylethoxycarbonyl)piperazin- I yl)aniline (0.868 g, 3.13 mmol), 4-hydroxybenzoic acid (0.525 g, 3.80 mmol), chloro-N,N,NI,N',-bis(pentainethylene)formnamidinium hexafluorophosphate (1.35 g, 3.74 inmol), and CH2Cl2 (45 mL). This mixture was cooled in an ice bath and diisopropylethylamnine (0.96 mL, WO 98/00134 PCT/US97/11133 -110- 5.52 mmol) was added. The ice bath was allowed to expire and the mixture was stirred at ambient temperature 18 h. The mixture was diluted with EtOAc and washed with 10% aqueous citric acid, water and brine. Drying (MgSO4), filtration and removal of the solvent in vacuo gave an oil. This material was chromatographed on 100 g of silica gel using 70% EtOAc-hexane as eluant to give 0.57 g of dimethylethoxycarbonyl)piperazin-1-yl)phenyl)-4-hydroxybenzamide as a grey solid.
IH NMR (CDCl3): 8 1.48 9H), 3.01 4H), 3.48 4H), 6.82 (d, J=9Hz, 2H), 6.85 j=9Hz, 2H), 7.58 j=8Hz, 2H), 7.81 j=8Hz, 2H), 9.80 (br s, 1H), 10.01 (br s, lH).
tert-Butyl -dimethylethoxycarbonyl)piperazin- 1yl)phenylaminocarbonyl)phenoxyacetate (14-5) To a 100 mL round bottomed flask with a stirring bar and an argon inlet was added -dimethylethoxycarbonyl)piperazin-1-yl)phenyl)-4-hydroxybenzamide (0.56 g, 1.41 mmol) DMF mL), Cs2CO3 (0.49 g, 1.51 mmol) and tert-butyl bromoacetate (0.28 mL, 1.75 mmol). This mixture was stirred at ambient temperature for 18 h. The mixture was diluted with EtOAc and washed with H20 and brine. Drying (MgSO4), filtration and removal of the solvent in vacuo gave 0.78 g of tert-butyl 4-(4-(4-(1,1-dimethylethoxycarbonyl)piperazin- 1 -yl)phenylaminocarbonyl)phenoxyacetate which was used in the next step without purification.
1 H NMR (DMSO-d6): 8 1.48 9H), 1.49 9H), 3.02 4H), 3.39 4H), 4.78 2H), 6.88 j=9Hz, 2H), 7.02 j=9Hz, 2H), 7.62 (d, j=8Hz, 2H), 7.82 j=8Hz, 2H), 9.82 (br s, 1H).
4-(4-(Piperazin-1-yl)phenylaminocarbonl)phenoxyacetic acid (14-6) To a 100 mL round bottomed flask with a stirring bar and a gas dispersion tube was added tert-butyl -dimethylethoxycarbonyl)piperazin-1-yl)phenylaminocarbonyl)phenoxyacetate (0.78 g, 1.52 mmol) and EtOAc (40 mL). This solution was cooled in an ice bath and HCI gas was sparged through the solution for 15 min. The WO 98/00134 PCT/US97/11133 resulting mixture was aged 90 min. The excess HC1 and EtOAc were removed in vacuo and the crude product was purified by preparative reverse phase HPLC. There was obtained 0.218 g of 4-(4-(piperazin- 1yl)phenylamninocarbonyl)phenoxyacetic acid as a crystalline solid.
1 H NMR (0.LIN NaOD-D20): 5 2.99 (in, 4H1), 3.15 (in, 4H), 4.5 9 (s, 2H), 7.03 j=9Hz, 2H), 7.16 j=9Hz, 2H), 7.7.37 j=8Hz, 2H), 7.88 j=8Hz, 2H).
WO 98/00134 WO 9800134PCTIUS97/11133 112 SCHEME 1. HO 3
S-&/N
2 +ClI
OH
H
3
C
H
3
O'
OH
3 2. Na 2
S
2
O
4 Boc 2 O CHC1 3 ref lux
CFH
3
NH
2 15-2
OH
H
3 0J OH 3 NHBoc Brl- COOEt 052003, DMF 0 COOEt 0 COQEt HOI (gas) EtOAc, 000
.HCI
OH
3
H
3 0 y CH 3 NHBoc
H
3
C
NH
2 15-4 WO 98/00134 WO 9800134PCT/UJS97/11 133 113 SCHEME 15 CONTINUED Boc-N CO CI PF6J
O
i-Pr 2 NEt, CH 2
CI
2 Boo-N N LiOH 1 COOEtDM,0 )M 2
OH
3
OCH
3
FCO
Boc-N N ri 15-7
CH
3 HOI (gas) EtOAc, 000 HN N-
.HCI
OH
3 1
-COOH
0
CH
3 15-8 WO 98/00134 WO 9800134PCTIUS97/11133 114- -Piperazinyl)phenylcarbonylamino)- 3 ,5 -dimethyl phenoxy)acetic acid, dihydrochioride (15-8) Using a -sequence essentially the same as described for compound 12-9, but starting with 3,5-dimethyiphenol (1 -piperazinyl)phenylcarbonylamino)-3 ,5-dimethylphenoxy)acetic acid, dihydrochioride was prepared, mp: >250'C.
I H NMR (D20): 5 2.15 3.32 (in, 411), 3.54 (in, 4H), 4.64 (s, 2H), 6.72 2H), 7.12 j=8.6, 2H), 7.17 (in, 2H), 7.83 j=8.6Hz, 2H).
WO 98/00134 WO 9800134PCTIUS97/11133 -115- SCHEME 16 I KMnO 4 /H 2 0 No -&C0 2
H
1. H 2 /PtO 2 /HOAc /MeOH psi 2. B0C 2 0
H
2 N O~O 2 Et
H
3 C 1
BOOND-O-'N
P F 6 i-Pr 2 NEt, 0H 2 C1 2 1. LiOH 2. HCI EtOAc 16-5 O,,,C02
H
H
3
C
WO 98/00134 WO 9800134PCTIUS97/11133 C0 2 H 16-2
I~-OCO
2 t CI PF 6 i-Pr 2 NEt, N NK&JCH 2
CI
2 10-4 N0 C H-P IC 2 Et
H
3
C
N 0 NI CO 2
H
16-7HC WO 98/00134 PCT/US97/11133 117- N- CO0 2
H
16-2 4-(4-Pyridinyl)benzoic acid (16-2) A slurry of 16-1 (5 g, 29.6 mmol, prepared as described in Chambron, J.C; Sauvage, Tetrahedron, 1987, 895 and Comins, Abdullah, J. Org. Chem., 1982, 47, 4315 method B) in 200 mL H20 was treated with 10% HCI until the solids dissolved. The solution was treated with solid KMnO4 in portions (11.2 g, 888 mmol), stirred until the KMnO4 hjad dissolved and heated to 90°C for 18hr.
An additional 2 g of KMnO4 was added and the reaction was again heated to 90°C for 2 hr. The reaction was cooled to ~60 0 C, filtered and the solids were washed with warm water. The filtrate was evaporated and the residue chromatographed (Silica gel, 10:1:1 EtOH/H20/NH40H) to give 16-2 as an off-white solid.
1H NMR (400 MHz, DMSO) 8 8.61 2H), 8.0 2H), 7.72 4H).
BOCN CO 2
H
16-3 4-(1 -(11 -Dimethylethoxycarbonyl)piperidin-4-yl)benzoic acid (16-3) A solution of 16-2 (0.5 g, 2.5 mmol) in 20 mL HOAc/MeOH was treated with 250 mg PtO2 and hydrogenated at 50 psi for 4 hr. The solution was filtered through Solka Floc, evaporated and azeotroped with heptane to remove excess HOAc. The intermediate amino acid acetic acid salt was obtained as a white solid.
Rf (10:1:1 EtOH/H20/NH40H) 0.3.
WO 98/00134PCUS/113 PCT/US97/11133 118 IH NMR (400 MHz, CD3OD) 8 8.96 (in, 2H), 7.35 (in, 2H), 3.5 (bd, 2h), 3.4 (mn, 2H), 3.2 (in, 2H), 3.0 (mn, 2H).
A slurry-of the amino acid (0.5 g, 1.9 mmol) in (12 m.L) was treated with 1 N NaGH (4.8 mL) and ditertbutyldicarbonate (0.564 g, 2.58 minol) at room temperature for 6 hr. The reaction was acidified to pH 5 with 10% KHS04 and extracted several times with EtOAc. The EtOAc layers were combined and evaporated to give 16-3 as a white solid.
Rf (97:3:1 CHCI3/MeOHIHOAc) 0.39.
IH NMR (400 MHz, CD3OD) 8 7.95 7.33 2H), 4.2 (bd, 2H), 2.85 3H), 1.8 (bd, 2H), 1.6 (mn, 2H), 1.48 9H).
N00 16-4 3 Ethyl 1-(1,1 -Dirnethylethoxycarbonyl)piperidin-4- .yl)p2henylcarbonvl aminio)-3-inethylphenoxv)acetate (16-4) A slurry of 16-3 (0.15g, 0.49 mimol) and 10-4 '(0.12g, 0.49 iniol) in CH2C12 was treated with chloro-N,N,N',N',bis(pentamethylene)formamidinium hexafluorophosphate (0.1 94g, 0.54 minol) and diisopropylethyl amine (0.34 niL, 1.96 mmol) and stirred at room temperature for 24 hours. The solution was diluted with EtOAc and washed with H20, 10% citric acid, saturated NaHCO3 and brine, dried over MgSO4, filtered and evaporated. The residue was chromatographed (silica gel 50% EtOAc/H-exanes) then triturated with ether/hexanes to give 16-4.
EtOAc/Hexanes) 0.33.
IH NMR (400 MHz,CDC13)5 7.81 (2s,2H), 7.70(d,1IH), 7.50 1 H), 7.31 (2s, 2H), 6.8 (mn, 2H), 4.6 2H), 4.28 2H), 2.8 (in, 2H), 2.72 (in, 1H), 2.30 3H), 1.85 (bd, 2H), 1.75 (mn,2H), 1.48 9H) 1.3 (t, 3H1).
WO 98/00134 PCT/US97/11133 -119- HN O H O.C O 2
H
H
3
C
2-(4-(4-(Piperidin-4-yl)phenylcarbonylamino)-3-methylphenoxy) acetate (16-5) A solution of 16-4 (0.1g, 0.2 mmol) in 1:1:1 was treated with LiOH (0.084g, 2 mmol) at room temperature. After 1 hour the reaction was diluted with EtOAc and KHSO4 and the layers were separated. The organic layer was washed with H20, brine, dried with MgSO4, filtered and evaporated to give the acid as a clear oil.
Rf(9:1:1 CH2CI2/MeOH/HOAc) 0.57.
1H NMR (400 MHz,CD30D) 8 7.94 (2s,2H), 7.44(2s,2H), 7.2 1H), 6.90 1H), 6.8 1H), 4.62 2H), 4.24 (bd, 2H), 2.9 2H), 2.83 1H), 2.28 3H), 1.86 (bd, 2H), 1.7-1.6 2H), 1.5 9H).
A slurry of the intermediate acid (0.9 g, 1.19 mmol) was cooled to -78 0 C and saturated with HCI gas. The reaction was warmed to 0°C, then concentrated in vacuo to give 16-5 as the HCI salt.
Rf (10:1:1 EtOH/H20/NH40H) 0.81.
1 H NMR (400 MHz, CD30D) 5 7.94 (2s, 2H), 7.44 (2s, 2H), 7.2 (d, 1H), 6.90 1H), 6.8 1H), 4.65 2H), 3.52 (bd, 2h), 3.15 (bt, 2H), 3.05 1H), 2.12 (bd, 2H), 1.97 2H).
WO 98100134 WO 9800134PCT[US97/11133 120 -H 1 6-6 Ethyl 2-(4-(4-(Pyridin-4-yl)phenylcarbonylamino)-3 -methylp2hen oxy) acetate (16-6) 16-2 (0.5 g, 2.5 mmol) and Ethyl 2-acetoxy-4-amino benzene hydrochloride (13 (0.579 g, 2.5 mmol) were coupled as described for 16-4 to give 16-6 after chromatography (silica gel, MeOHEthOAc) followed by reverse phase preparative HPLC.
IH NMR (400 MHz, CD3OD) 5 8.82 2H), 8.30 2H), 8.15 2H), 8.05 2H), 7.60 2H), 6.98 2H), 4.70 2H1), 4.23 2H), 1.30 3H).
ND
N
H QC 2
H
2-(4-(4-(Pvridin-4-yl)p2henylcarbonylaminophenoxy)acetic acid (16-7) 16-6 was treated with LiOH as described for 16-5 to give 16-7 after chromatography (silica gel, 10: 1:1. EtOHIH2O/NH4OH) followed by preparative reverse phase HPLC.
Rf (10: 1: 1 EtOH/1-20/NH4OH) 0.76.
1 H NMR (400 MHz, DMSO) 8 8.65 2H), 8. 10 2H), 7.96 2H), 7.8 2H), 7.65 2H), 6.90 2H), 4.58 (bs, 2H).
WO 98/00134 WO 9800134PCT/US97/11133 121 SCHEME 17 N C H 3 16-1 INaNH 2 Di methylani line 1600 C H 3 17-1
H
2
N
A Cl,,-C O N\ C H3 17-2 tBuO NH
AIBN
Br tBuO NH
B
WO 98/00134 WO 980134PT/UJS97/11 133 122 SCHEME 17 (CONT'D) 17-3 SAgNO 3
THF/H
2 0 0 o N\
H
tBuO NH INaCIO 2
H
2 0 2
CH
3
CN
0 ON OH
Z
tBuO NH PYCLU, Et 2 NiPr HCI* H 2 N Ol-COOEt
CHCI
1 3-9 0 tBuO J NH H-&17-6 LiOH, THF/H 2 0/MeOH WO 98/00134 PTU9/13 PCTfUS97/11133 123 SCHEME 17 (CONT'D) tBuO NH N N-IO H 17 H \/O-OH l HCI/EtOAc 0
H
2 N a 17-8 WO 98/00134 PCT/US97/11133 -124- N\
CH
3
H
2
N
1 1 2-Amino-4-(4-methvlphenyl)pyridine (17-1) NaNH2 (142 mmol) was prepared in situ as described in Leffler, M.T. Organic Reactions 1942 Vol. I. Dimethylaniline (20 ml) was added dropwise to the freshly prepared NaNH2. 16-1 (8.0 g, 47.3 mmol) was dissolved in a small amount of dimethylaniline and added to the mixture. The slurry was heated to 1600 for 2 hours. The reaction mixture is cooled and diluted with 10% KHSO4 and water. The layers are separated, and the dimethylaniline layer is washed with KHSO4. The aqueous layers are combined, basified with saturated NaHCO3, and extracted with EtOAC The combined organic layers are washed w/brine and concentrated to yield an oily solid. This oily solid was purified by flash chromatography MeOH/EtOAc) to give 17-1 as a tan solid. Rf MeOH/EtOAc) 0.36. 1 H NMR (400 MHz, DMSO-d6) 8 7.9 1H), 7.5 2H), 7.3 2H), 6.7 1H), 6.6 1H), 5.9 1H), 2.3 3H).
ON CH 3 tBuO NH 17-2 2-(1,1 -Dimethylethoxycarbonylamino-4-(4-methylphenyl) pyridine (17-2) 17-1 is dissolved in dichloroethane (30 mL) and added dropwise, rapidly to a refluxing solution of Boc20 in dichloroethane mL). After the addition, the reaction is refluxed for 30 min, then stirred at RT overnight. The reaction mixture is cooled and WO 98/00134 PCT/US97/11133 -125concentrated to yield a yellow solid, which is purified by chromatography (silica, 30% EtOAc/Hexane) to give 17-2 as a white solid. Rf (50% EtOAc/Hex) 0.73. 1 H NMR (400 MHz, CDC13) 6 8.3 IH), 8.2 1H), 7.5-7.6 4H), 7.3 (bs, 1H), 7.1 1H), 2.3 (s, 3H), 1.5 9H) Br ON Br tBuO NH 2-(1,1-Dimethylethoxycarbonylamino-4-(4-dibroomethylphenyl) pyridine (17-3) 17-2 (1.0 g, 3.5 mmol) is dissolved in CC14. NBS (1.3 g; 7.3 mmol) is added, followed by AIBN (28.7 mg, 0.18 mmol). The reaction is heated to reflux while shining a white light directly on the flask. After 2 hours, AIBN (28.7 mg) is added, the light is turned off, and the reacion is stirred overnight to effect completion to product.
The reaction mixture is cooled and the succinimide is filtered off, washing the solids with CCl4. The filtrate is concentrated to give 17-3 as a brown solid which is used in the next step without further purification. Rf (50% EtOAc/Hex) 0.70. 1 H NMR (400 MHZ, CDC13) 6 8.3 1H), 8.2 IH), 7.7 3H), 7.2 (bs, 1H), 7.1 1H), 6.7 (s, 1H), 1.5 (s,9H).
WO 98/00134 PCT/US97/11133 -126- ON H 0 tBuO NH 17-4 -Dimethylethoxycarbonylamino)pyrid-4-yl) benzaldehvde (17-4) 17-3 (1.5 g, 34 mmol) is dissolved in water/THF (15 mL). AgNO3 (1.1 g, 6.8 mmol) is added and stirred for 1/2 hour. The reaction mixture is diluted w/EtOAc and water. The layers are separated and the organic layer is filtered to remove the solids. The filtrate is washed w/brine, dried (MgSO4), filtered and concentrated to yield a brown solid. The solid is purified on a "plug" of silica, eluting first w/50% EtOAC/hexane, then 100% EtOAc to elute all of the product. The fractions are concentrated to give 17-4 as a brown solid.
Rf (50% EtOAc/Hexane)0.62. 1 H NMR (400 MHz, CDC13) 8 10.1 (s, 1H), 8.29 8.26 8.0 7.8 2H), 7.5 (bs, 1H), 7.2 1.5 9H).
ON
n'J OH tBuO NH 1 17-5 1,1 -Dimethylethoxycarbonylamino)pyrid-4-yl) benzoic acid (17-5) 17-4 (1.0 g, 3.4 mmol) is slurried in CH3CN (40 mL).
KHSO4 (1.3 mL) and 30% H202 (0.957 mL) are added and the solution is cooled in an ice bath. NaC102 (8.5 mmol, 770 mg) is added dropwise in 50 mL water over 20 min. The reaction is stirred for 16 hrs. The reaction mixture is diluted w/EtOAc and washed with saturated NaHCO3 The aqueous layers are combined, acidifed WO 98/00134 WO 9800134PCTIUS97/1 1133 127 with 10% KHSO4, and extracted w/CH2CI2 (2x) and EtOAc (2x) to give 17-5 as a yellow solid. IH NMR (400MHz, DMSO-d6) 8 12.6-12.8 (bs, I1H), 9.9 I 8.3 1d 1H), 8.0-8.1 (in, 3H), 7.9 2H), 7.3 I H), 9H).
0 ON/ N 0 '-COEt tBu0 NH H 17-6 Ethyl 1,1-Dimethylethoxycarbonylamino)pyrid-4yl)carbonylamino)p2henoxy)acetate (17-6) A mixture of 17-5 (300 mg, 0.956 mmol), 13-9 (0.956 mmol, 236 mg), diisopropylethylamine (3.8 inmol, 0.66 mL), PYCLU (1.05 mmol, 378 mg), and CH2CI2 (10 ml) is stirred at RT for 16 hours. The reaction mixture is diluted with EtOAc and washed with saturated NaHCO3 and Brine. The Organic layer is dried (MgSO4), filtered and concentrated to yield a pink solid. Triturations with hexanes followed by flash chromatography (silica, 30% EtOAc/Hex) affored 17-6 as a white solid. Rf (30% EtOAc/Hexanes) 0.24. 1H NMR (400 MHz, CDC13) 8 8.3 1H), 8.2 1H), 7.9 2H), 7.8 (2d, 3H), 7.4 (bs, I1H), 7.6 (bs, ILH), 7.2 I1H), 6.8 I1H), 6.7 11-H), 4.6 (s, 2H), 4.3 2H), 2.3 3H), 1.5 9H), 0.9 3H).
WO 98/00134 WO 9800134PCT[US97/11133 128 0 ON/ N 0 tBuO NH 1,1 -Dimethylethoxycarbonylamino)pyrid-4vl)carbonylamino)p2henox') acetic -acid (1 7-7') A mixture of 17-6 (0.17 mmnol, 90 mg), LiOH (0.60.mmol, mg), and THF/H20/MeOH/ (1 mL/1 mnL/1 mL) is stirred for 2 hours. The reaction mixture is diluted with EtOAc and 10% KHSO4.
The aqueous layer is back-extracted with EtOAc. The organic layers are combined, washed with brine, dried (MgSO4), filtered and concentrated to yield 17-7 as a white solid. 1 H NMR (400 MHz, CD3OD) 8 8.3 1H), 8.1 (bd, 3H), 7.9 2H), 7.5 (bs, 1H), 7.2 (s, 1H), 6.9 1H), 6.8 LH), 4.7 2H), 2.3 3H), 1.5 9H).
0 N C00H
H
2
NH
2-(4-(4-(2-Aminopyrid-4-yl)carbonylamino)phenoxy)acetic acid (17-8) 17-7 (0.14 mmol, 70 mg) is dissolved in EtOAC (3 mL) and cooled to HCl is bubbled throu gh until the solution is saturated. The reaction is stirred at 0' for 3 hours, then at 350 for 16 hours. The reaction mixture is concentrated to yield a tan solid, which is purified by flash chromatography (10/0.25/0.25 EtOH/NH4OH/H20) to yield 17-8 as an off white solid. Rf (10/0.510.5/0.5 EtOH/NH4OH/H20) 0.86. 1 H NMR (400MHz,-DMSO-d6) 8 9.8 (s, LH), 8.0 2H), 7.9 lH), 7.7 2H), 7.2 IH), 6.9 (in, 2H), 6.8 (mn, 2H), 6.0 lH), 2.2 3H).
WO 98/00134 WO 98/0 134PCTIUS97/11 133 129 SCHEME 18 -Na
NH
2
ON
18-1 aq. H 2 S0 4 /dioxane
CH
3 0H 0
OCH
3 WO 98/00134 WO 9800134PCTIUS97/11133 -130- SCHEME 18 (CONT'D) 18-2 I1.) H 2 10% Pd/C Boc 2 O, Et 3
N
0 0 Na OCH 3 tBuO
H
18-3 1N NaOH .O1 11C0 2
E
10-4
H
2
N'
PYCLU, DIPEA 0 tBuO
CH,
3 HCI (g) LiOH e-"0 ll-C0 2
H
H
OH
3 1 WO 98100134 WO 9800134PCTIUS97/11133 131
H
4-(3(R)-((Nl-benzyl)p2yrrolidinyl)amino)phenvlnitrile (18-1) A solution of N-benzyl-3-(R)-aminopyrrolidine (TCI, 0.45 g, 2.27 mmnol) in 1 mL acetonitrile was treated with 4-fluorophenylcyanide (Aldrich, 3 g, 25.5 mnmol) and heated to 100 0 C. The acetonitrile was allowed to evaporate and the slurry was heated for 18 hours. Additional 4-fluorophenylcyanide (1 g) was added and the reaction heated for 18 hours. The oily mixture was absorbed onto silica gel and eluted first with 20% EtOAc/Hexanes, then with MeOH/CHC13 saturated with NI-3 to give 18- 1.
Rf MeOH/CHCI3 saturated with N113) 0.55.
III NMR (400 MHz, CDCI3) 8 7.4 2H), 7.3 (in, 5H), 6.5 2H), 4.45 (bd, 1H), 4.0 (in, 1H), 3.64 211), 2.83 (in, 1H), 2.75 (dd, 1H), 2.6 (dd, 1H), 2.43(mn, 1H), 2.34 (in, i1H), 1.6 (in, iR).
0 00H 3
H
Methyl 4-(3(R)-((N-benzyl)p2yrrolidinyl~amino)benzoate (18-2) A solution of 18-1 (0.52 g, 1.87 inmol) in dioxane (10 mEL) and 1: 1 H120/concentrated H2S04 (10 mL) was heated to 1 00 0 C for 18 hours. The solvent was removed in vacuc and the residue dissolved in MeOH (30 mL) and stirred for 4 days. The solvent was removed and WO 98/00134 PCT/US97/11133 -132the residue taken up in saturated NaHC03 and 1N NaOH to give a pH of then extracted with chloroform. The organic extracts were combined, evaporated and the residue chromatographed in a gradient of 20-30% EtOAc/Hexanes to give 18-2 as a clear oil.
Rf=(50% EtOAc/Hexanes) 0.31.
1H NMR (400 MHz, CDC13) 8 7.45 2H), 7.30 5H), 4.35 (bd, 1H), 4.05 IH), 3.34 3H), 3.63 2H), 2.8 2H), 2.58 (dd, IH), 2.45 (dd, 1H), 2.84 1H), 1.65 1H).
0 0 N
OCH
3 tBuO H 18-3 Methyl -dimethylethoxycarbonyl)pyrrolidinyl) amino)benzoate (18-3) A suspension of 18-2 (0.29 g, 0.93 mmol) and 10% Pd/C (0.068 g, 23% by weight) in CH30H (7 ml) was stirred under hydrogen atmosphere for 48 hours. The mixture was filtered through Solka Floc, and solvent removed. The residue was dissolved in CH2C12 (5 ml) and treated with Et3N (0.65 ml, 4.7 mmol) and BOC20 (0.45 g, 2.1 mmol).
The reaction mixture was stirred 1 h at room temperature, then diluted with EtOAc and washed with 10% KHSO4 and brine, dried (Na2SO4), and the solvent removed. The residue was purified by flash chromatography on silica gel, eluting with EtOAc (1)/hexane to provide 18-3 as an oil.
Rf=0.3 (silica gel, EtOAc (3)/hexane 1 H NMR (400 MHz, CDC13) 8 1.47 (9H, 1.92 (1H, 2.20 (1H, q), 3.25 (3H, 4.11 (IH, 4.19 (1H, 6.55 (2H, 7.87 (2H, d).
WO 98/00134 PCT/US97/11133 133- 0 O
CO
2 Et O N N i tBuO
N
H
18-4 Ethyl 2-(3-methyl-4-(4-(3-(R)-((1-(1,1-dimethylethoxycarbonyl)pyrrolidinyl)amino)phenylcarbonvlamino)phenvloxy)acetic acid (18-4) A solution of 18-3 (0.245 g, 0.76 mmol) in dioxane (4 ml) was treated with IN NaOH (3.0 ml, 3.0 mmol) at room temperature for 1 h. A second portion of IN NaOH (3.0 ml, 3.0 mmol) was then added, and the reaction mixture was stirred for 15 h. The solution was then adjusted to pH 7 with IN HCI and the solvent removed. The residue was suspended in CH2C12 (6 ml) and treated with 10-4 (0.19 g, 0.77 mmol), diisopropylethylamine (0.53 mmol, 3.0 mmol), and PYCLU (Fluka, 0.30 g, 0.83 mmol). The reaction mixture was stirred for 48 h at room temperature, then solvent was removed and the residue was purified by flash chromatography on silica gel, eluting with EtOAc (3)/hexane (2) to give 18-4 as an oil.
Rf=0.2 (silica gel, EtOAc (3)/hexane 1 H NMR (400 MHz, CDC13) 8 1.30 (3H, 1.47 (9H, 1.93 (1H, s), 2.24 (1H, 2.28 (3H, 3.27 (1H, 3.49 (2H, 3.73 (1H, m), 4.11 (2H, 4.16 (1H, 4.29 (2H, 4.60 (2H, 6.62 (2H, d), 6.77 (1H, 6.81 (1H, 7.41 (IH, 7.68 (IH, 7.73 (2H, d).
WO 98/00134 WO 9800134PCT/US97/11133 -134- HN>, Hq N&
CH
3 H 1- 2-(3-Methyl-4-(4-(3 (R)-(pyrrolidinyl)amino)phenylcarbonylamnino) phenvloxv)acetic acid (18-5) A solution of 18-4 (0.216 g, 0.434 mmol) in EtOAc ml) was cooled to -78'C and treated with HCI for 1.5 min. The reaction mixture was stirred at 0 0 C for I h and then solvent was removed. The residue taken up in THF (1)/CH3OH (1)/H20 (6 mnl) and treated with LiOH*H20 (0.1 1g, 2.6 mmnol) at room temperature for h. The solvent was removed and the residue was purified by flash chromatography on silical gel eluting with EtOH (1 8)/H20 (1 )INH4OH to afford 18-5 as a white solid.
Rf=-0.2 (silica gel, EtOH (18)/H20 (1)INH4OH 1 H NMR (400 MHz, D20) 8 1.57 (1 H, in), 2.06 (1 H, in), 2.08 (3H, s), 2.60 (1H, in), 2.76 (1H, in), 2.86 (lH, in), 3.03 (lH, in), 3.93 (1H, in), 4.38 (2H, 6.71 (3H, in), 6.80 (1H, 7.05 (1H, 7.66 (2H, d).
WO 98/00134 WO 9800134PCT/US97/11133 -135 SCHEME 19 H2N".- 9 N- Boc 2 O, Et 3
N
HI
tBuO-
H
2
ON
a H "N J
C
0 19-2 WO 98/00134 WO 9800134PCT/US97/11133 136 SCHEME 19 (CONT'D) 19-2 aq. H 2 S0 4 dioxane
CH
3 0H BoC 2 O, Et 3
N
0 'N OCH 3 Hb tBuO-\ 1 -i 0 11.) 1N NaOH 10-4 PYCLU, DIPEA 0 0 N-C0 2 Et AN 19-4 H 'qH. 3
H
N
tBuO- 0 HCt (g) LiOH 19-5
OH
3 WO 98/00134 PCT/US97/11133 137-
H
tBuO -if N& C N 19-1 N-benzvl-3-(R)-(( 11 -dimethlethoxycarbonyl)amino)pyrrolidine (19-1) A solution of N-benzyl-3-(R)-aminopyrrolidine (TCI, 5.00 g, 28.4 mmol) in CH2CI2 (100 ml) was treated with Boc20 (6.80 g, 31.2 mmol) and Et3N (7.9 ml, 57 mmol). The reaction was stirred at room temperature for 15 h, then solvent was removed. The residue was purified by flash chromatography on silica gel eluting with EtOAc (3)/hexane to give 19-1 as an oil.
Rf=0.4 (silica gel, EtOAc hexane 1H NMR (400 MHz, CDC13) 8 1.43 (9H, 1.60 (1H, 2.27 (2H, m), 2.51 (1H, 2.62 (1H, 2.76 (1H, 3.58 (2H, 4.13 (1H, m), 4.89 (1H, 7.29 (5H, m).
H
tBuO N O Y N
CON
19-2 1,1 -Dimethylethoxycarbonyl)amino)pyrrolidin-1 vl)phenylnitrile (19-2) A suspension of 19-1 (7.43 g, 26.9 mmol) and 10% Pd/C (2.45 g, 33% by weight) in CH30H (150 ml) was stirred under H2 (g) atmosphere for 5 h. The reaction mixture was filtered through a pad of Solka Floc, and the solvent was removed. The residue was dissolved in CH3CN (20 ml) and treated with 4-fluorobenzonitrile (16.1 g, 133 mmol) at 70 0 C for 18 h. The reaction mixture was then cooled to room temperature and the solvent was removed. The residue was purified by WO 98/00134 PCT/US97/11133 138flash chromatography on silica gel eluting with EtOAc (1)/hexane to afford 19-2 as a white solid.
Rf=0.4 (silica gel, EtOAc (2)/hexane 1 H NMR (400 MHz, CDC13) 8 1.45 (9H, 1.99 (1H, 2.30 (1H, m), 3.18 (1H, 3.46 (2H, 3.62 (1H, 4.36 (1H, 4.67 (1H, b), 6.52 (2H, 7.47 (2H, d).
H
tBuO N o 0
OCH
3 19-3 Methyl 4-((3-(R)-(1,1-dimethylethoxycarbonyl)amino)pyrrolidin-1yl)-benzoate (19-3) A solution of 19-2 (1.48 g) in dioxane (25 ml) and H2S04 (1)/H20 (25 ml) was heated at 70 0 C for 15 h. The reaction mixture was then concentrated to remove the dioxane, diluted with CH30H (100 ml), heated at 60 0 C for 2 h, and stirred at room temperature for 48 h.
The reaction mixture was concentrated to remove the CH30H, and the residue adjusted to pH 10 with 6N NaOH and sat. NaHCO3. The aqueous mixture was then extracted with CH2C12, and the organic extracts concentrated to afford an oil. This residue was dissolved in CH2C12 (25 ml) and treated with Boc20 (2.96 g, 13.6 mmol) and Et3N (2.2 ml,. 16 mmol). The reaction mixture was stirred at room temperature for 15 h. The solvent was then removed and the residue purified by flash chromatography on silica gel eluting with EtOAc (2)/hexane to yield 19-3 as an oil.
Rf=0.5 (silica gel, EtOAc hexane IH NMR (400 MHz, CDC13) 8 1.27 (9H, 1.99 (1H, 2.30 (1H, m), 3.22 (1H, 3.42 (2H, 3.85 (3H, 4.37 (1H, 4.69 (1H, b), 6.51 (2H, 7.91 (2H, d).
WO 98/00134 PCT/US97/11133 139-
H
tBuO YN H
CO
2 Et 19-4
CH
3 Ethyl -dimethylethoxycarbonyl)amino)pyrrolidin- 1-yl)phenylcarbonvlamino)-3-methylphenoxv)acetic acid (19-4) A solution of 19-3 (0.418 g, 1.30 mmol) in dioxane (10 ml) was treated with IN NaOH (7.0 ml, 7.0 mmol), and the reaction mixture was stirred at room temperature for 15 h. A second portion of IN NaOH (3.0 ml, 3.0 mmol) was then added and the reaction mixture was stirred an additional 15 h. Solvent was removed, and the residue was taken up in H20, adjusted to pH 6-7 with 10% KHSO4 and extracted with EtOAc. The combined organic extracts were dried (Na2SO4) and the solvent was removed. The residue was taken up in CH2C12 (7 ml) and treated with 10-4 (0.33 g, 1.3 mmol), DIPEA (0.81 mmol, 4.6 mmol) and PYCLU (Fluka, 0.53 g, 1.5 mmol). The reaction mixture was stirred at room temperature for 48 h. The solvent was then removed and the residue was purified by flash chromatography on silica gel eluting with EtOAc (1)/hexane to afford 19-4 as a white solid.
Rf=0.3 (silica gel, EtOAc (1)/hexane 1H NMR (400 MHz, CDC13) 8 1.30 (3H, 1.46 (9H, 2.04 (1H, m), 2.29 (3H, 2.31 (1H, 3.23 (1H, 3.45 (2H, 3.65 (1H, m), 4.27 (2H, 4.38 (1H, 4.60 (2H, 4.72 (1 H, 6.57 (2H, d), 6.77 (1H, 7.67 (1H, 7.41 (1H, 7.72 (1H, 7.77 (2H, d).
WO 98/00134 WO 9800134PCTIUS97/11133 -140- 0 OIS CO 2
H
N N N I H
CH
3 19-5 -amino)pyrrolidin- 1 -yl)phenylcarbonylamino)-3 methylphenoxv)acetic acid (19-5) A solution of 19-4 (0.306 g, 0.615 mmol) in EtOAc ml) at -78'C was treated with HCl for 1.5 min. The reaction mixture was then stirred at 0 0 C for 1 h, and the solvent was removed.
The residue was taken up in THF (1)/CH3OH(1)/H20 (6 ml) and treated with LiOH*H20 at room temperature for I h. The solvent was removed, and the residue was purified by flash chromatography on silica gel eluting with EtOH (18)/H20 (1)INH4OH to give 19-5 as a white solid.
Rf=-0.4 (silica gel, EtOH I 8)/.H20 (1)INH-40H.
1H NMR (400 MHz, D20) 6 1.74 (IH, in), 2.10 (3H, 2.12 (IH, in), 3.00 (1 H, in), 3.28 (1 H, in), 3.39 (1 H, in), 3.47 (1lH, mn), 3.59 (1 H, mn), 4.39 (2H, 6.61 (2H, 6.71 (iIH, 6.81 (1H, 7.07 (LIH, 7.79 (2H, d).
WO 98/00134 WO 9800134PCT/US97/11133 -141- SCHEME 0 N -NC O'Li+ IMeOH/HOAC conc. HCI H 2 0/PtO 2
/H
2 0 HOAc HND -NC K OH 20-1 I N NaOH/H 2 0/Dioxane 0 0 tBuOANa N OH 2- PYCLU HCI H 2 N 0 COOCH 2
CH
3
CH
2 01 2 DIPEA OH 3 IQ0-4 0 0 tBuO 'kN N N NICB -iD 20-3
CH
3 WO 98/00134 WO 9800134PCT[US97/11133 -142- SCHEME 20 (CONT'D) 20-3 1) LIGH
THF/H
2 0/MeOH 2) 1N HCt O01-.COOH 20-4
CH
3 HCI (g) EtOAc N 0 O COOH
OH
3 -4 Su- _o WO 98/00134 PCT/US97/11133 -143- 0 HOAc*HN N 'OH 20-1 4-(Piperidin-4-vl)piperidinvl carboxylic acid, acetic acid salt (20-1) A suspension of the lithium salt of N-(pyridin-4-yl)-4piperidine carboxylic acid (prepared as described in Tetrahedron, 1988 44(23) 7095-7108, 1 g, 5.55 mmol) in 50 mL 20% HOAc/MeOH, mL H20 and 10 mL concentrated HCI was treated with Pt02 (1.7 g) and hydrogenated at 65 psi for 24 hrs. The solution was filtered, evaporated and the residue azeotroped with heptane to yield 20-1 as a white solid.
Rf(9:1:1 EtOH/H20/NH40H)=0.39.
1H NMR (400 MHz, D20) 8 3.6-3.5 4H), 3.9 1H), 3.1-3.0 (m, 4H), 2.65 1H), 2.32 2H), 2.25 2H), 2.0-1.7 4H).
O 0 tBuO N N OH -Dimethylethoxycarbonyl)piperidin-4-yl)piperidinyl carboxylic acid (20-2) A solution of 20-1 (1.7 g, 5.55 mmol) in H20 (10 mL) was treated with sufficient 1N NaOH to bring the solution to pH 11.
Dioxane (20 mL) was added followed by BOC anhydride (1.33 g, 6.2 mmol) dissolved in 5 mL dioxane. After 1 hr the solvents were removed and the residue was triturated with acetonitrile and n-butanol, filtered and the filtrate concentrated. The residue was chromatographed (silica gel, 9:1:1 EtOH/H20/NH40H) to give 20-2 as a white solid.
Rf(9:1:1 EtOH/H20/NH40H)=0.70 WO 98/00134 WO 9800134PCT/US97/11133 144- 1 H NMR (400 MHz, D20) 8 4.12 (bd, 2H), 3.5 (bd, 2H), 3.32 (in, I1H), 2.95 (bt, 2H), 2.75 (bt, 2H), 2.9 (in, 1H), 2.15 (bd, 2H), 2.0 (bd, 2H), 1.7 (mn, 2H), 1.55 (in, 2H), 1.35 9H).
0 0 tBuO No N" N _Q 0 _C00Et 20-3
CH
3 Ethyl 1,1 -diinethylethoxycarbonyl)piperidin-4yl)piperidinylcarboxamido)3 -methylp~henoxy)acetic acid (20-3) (486 mg; 1.6 inmol) and 10-4 (250 mg, 1.6 minol) were dissolved in CH2C12. PYCLU (1.8 inmol, 634 mg) was added, followed by dilsopropylethylamine (6.4 ininol, 1.1 inL). The reaction is stirred for 2 days. The reaction mixture is diluted with EtOAc and washed with H20, 10% KHSO4, sat NaHCO3 and brine. The organic layer is dried (MgSO4), filtered and concentrated to yield a tan oil.
Flash chromatography (10% MeOH/CHCl3 sat N-H3) yielded desired product (20-3) plus bis piperidine urea by-product. This material is triturated with hexane to remove most of the by-product.
Rf (10% MeOH/ CHC13 sat. NH3)=0.20 I H NMR (400 MHz, CdCl3) 8 7.55-7.57 IH), 6.88 lH), 6.74 (s, 1H) 6.7 1-6.72 LH), 4.6 1H), 4.27-4.29 2H), 4.23-4.25 (bs, 2H), 2.9-3.0 2H), 2.6-2.7 2H), 2.35-2.45 2H).
WO 98/00134 WO 9800134PCTIUS97/11133 145- 0 0 tBu0'N- ND-- N -Q 0I-CO0H 21A4 CH 3 1, 1 -Dimethylethoxycarbonyl)piperidin-4-yl)piperidinylcarboxamido)3-methylphenoxy)acetic acid (20-4) (300 mg) was dissolved in THF/H20/MeOH (2 rnL/2 mL/2 m1L). LIOR (50 mg) was added and the reaction is stirred for 2 hours. IN HCl was added (0.50 mL) and the reaction mixture was concentrated to yield 20-4 as a tan solid.
Rf(9/1/1 CH2CI2/MeOHIHOAc)=0.16 IH NMR (400 MHz, CD3OD) 8 7.06-7.08 6.81 1H), 6.75 (d, 1H), 4.88 2H), 4.12-4.15 (bd, 2H), 3.04-3.07 (bd, 2H), 2.6-2.8 (vbs, 2H), 2.25-2.45 (in, 4H), 2.17 3H), 1.46-1.56 (in, 6H), 1.45 (9H).
0 HNaNa N\ ONI-C00H
CH
3 2- 2-(4-(4-(Piperidin-4-yl)piperidinylcarboxamido)3-methylphenoxy) acetic acid (20-5) 20-4 (0.08 mmol) is dissolved in EtOAc and cooled to -78'.
HCl is bubbled through until the solution is saturated. The reaction is warmed to and stirred for five minutes. The reaction mixture is concentrated to yield a tan solid.. The material was purified by flash chromatography using 10/1/1 EtOH/NI-40H/H20 as an eluent, resulting in pure 20-5 as a white solid.
Rf(I0/l/l EtOHINH4OH/H20)=0.15 WO 98/00134 PCT[US97/11133 -146- I H NMR (400 MHz, D20) 7.0 I1H), 6.8 I 6.7 (dd, I1H), 4.7 (s, 1H), 3.4 2H), 3.1 (bd, 2H), 2.9 2H), 2.8 (bt, IH), 2.5 (bs, 3H), 2.10-2.13 2H), 2.06 3H), 1.96-1.99 2H), 1.65-1.75 (in, 4H).
WO-98/00134 PTU9/13 PCTIUS97/11133 -147- SCHEME 21 0
CH
3 1:3
CH
3 a OH 21 Pyridine 12 NaOH 0 0 HO +HO
OH
3 OH OH 3 0/o Pd/C
H
2 MeOH 0
HO
OH
3
OH
21-3 SMeOH
CH
3 0
OH
3
OH
21-2 21-4 WO 98/00134 WO 98/0013 PCT1JS97(11 133 148 SCHEME 21 (CONTD) BrCH 2 C00Et 0S 2 00 3
DMF
:IO,,-CO
2 Et 21-5
CH
3
NBS
CC1 4
A
0
CH
3 Ort Br 0--CO 2
E
21-6 tBuO N r\N \NH 2 14-3
A
CAH
tBuO N N N
NZ
.7 0 00 2 Et 21.
WO 98/00134 WO 9800134PCTIUS97/11133 149 SCHEME 21 (CONTD) 21-7 U LOH ITHF/H 2 0/MeOH 0 0 tBuO N N N \O SHCI(g); EtOAc C2 H PLC 0 TFA -HN N 21-9 WO 98/00134 PCT/US97/11133 150- 0
HO
CH
3
OH
21-3 4-Hvdroxy-2-methyl benzoic acid (21-3) 21-3 was prepared in two steps from 4'-hydroxy-2'methylacetophenone, 21-1, (Aldrich, 18.0 g, 0.120 mol).
The first step employed the procedure found in Merck Process Patent US 852870-92031, which resulted in a 70/30 mix of 21-3 and 3-iodo-4-hydroxy-6-methyl benzoic acid (21-2) For the second step, the mixture (7.0 g) of 21-2 and 21-3 was reduced in MeOH (200 mL) using 10% Pd/C (700 mg) under a hydrogen balloon. After 16 hours of stirring, the reaction mixture was filtered through celite, the celite pad was washed with MeOH and the filtrate was concentrated to yield pure 21-3 as a tan solid.
1H NMR (400 MHz, CDC13) 5 7.8 1H), 6.6 2H), 2.5 3H).
Rf (90 Ethyl Acetate/Hexane)=0.34 0
CH
3
OO
CH
3
OH
214 Methyl 4-hydroxv-2-methylbenzoate (21-4) 21-3 (5.0 g, 32.9 mmol) was redissolved in MeOH (150 mL) and cooled to 00. HCI was bubbled through for a few minutes, the cooling bath was removed and the reaction was stirred at RT for 16 WO 98/00134 PCT/US97/11133 151 hours. The reaction mixture was then concentrated to yield pure 21-4 as a tan solid.
Rf (50% Ethyl Acetate/Hexane)=0.83 1 H NMR (400 MHz, CDC13) 5 7.9 IH), 6.7 2H), 3.85 3H), 2.57 3H).
0
CH
3 0
CH
3 O COOEt 21-5 Ethyl 2-(3-methyl-4-methoxvcarbonylphenoxv)acetate (21-5) 21-4 (1.5 g; 9.0 mmol) was dissolved in DMF (150 mL).
Ethyl bromoacetate (9.0 mmol, 1.0 mL) was added followed by Cs2CO3 mmol, 2.9 The slurry was stirred vigorously for 16 hours.
The reaction mixture was diluted with EtOAc and washed twice with water, 10% KHSO4 and brine. The organic layer was dried (MgSO4), filtered and concentrated to yield 21-5 which will be used in the next step without any further purification.
Rf (30 Ethyl Acetate/Hexane)=0.40 1 H NMR (400 MHz, CDC13) 8 7.9 1H), 6.7-6.8 2H), 4.6 2H), 4.3-4.3 2H), 3.9 3H), 2.6 3H), 1.3 3H).
WO 98/00134 PCT/US97/11133 152- 0
.CH
3 aO Br--O COOEt 21-6 Ethyl 2-(3-bromomethyl-4-methoxycarbonylphenoxy)acetate (21-6) 21-5 (2.1 g; 8.3 mmol) was dissoved in CCl4 (40 mL).
NBS (10.3 mmol, 1.9 g) was added followed by dibenzoyl peroxide (1.66 mmol, 400 mg). The reaction mixture was heated to reflux for 16 hours. The reaction mixture was cooled and filtered through a celite pad to remove excess succinimide. The filtrate was concentrated to yield 21-6 as a tan oil, which was used in the next step without any further purification.
Rf (30% Ethyl Acetate/Hexane)=0.52 O
O
tBuO N N N COOEt 21-7 Ethyl -Dimethylethoxycarbonylpiperazin-1 -yl)phenyl)-2-oxo- (21-7) 21-6 (8.3 mmol) was dissolved in benzene (100 mL), followed by 14-3. The mixture was heated to reflux for 32 hours.
Triethylamine (8.3 mmol) was added and the reaction was refluxed for 2 more days. The reaction mixture was then concentrated to yield a brown solid which was absorbed on silica and purified by flash chromatography. A small amount of the product was eluted with EtOAC/Hex. Most of the material was then flushed off the column with WO 98/00134PCIS/113 PCTIUS97/11133 153 MeOH/CHCI3 saturated with N113. These flushings yielded a brown solid which was repurified on a column of silica using MeOH/CHCl3 as a eluent to yield 21-7 as a brown solid.
Rf (50% EtOAc/Hex)=0.48 1 H NMR (400 MHz, CDCl3) 8 7.8 I1H), 7.7 2H), 7.0 (in, 4H), 4.77 2H), 4.72 2H), 4.3 2H), 3.6 (in, 4H), 3.1 (in, 4H), 1.5 (s, 9H), 1.3 3H).
00 tBu 0 N N N 1 1 -Dimethylethoxycarbonylpiperazin- 1 -yl)phenyl)-2-oxo- -oxy) acetic acid (21 -8) 21-7 (1.3 g; 2.6 mmol) was dissolved in THFIH2O/MeOH mL/5/5). LiOH (5.2 inmol, 218 ing) was added and the reaction mixture was stirred for 2 hours. 1 N HCI was added (2.6 and the reaction mixture was concentrated to yield 21 -8 and excess LiCI as a brown solid.
Rf(9/1/1 CH2CH2IMeOH/H1OAc)=-0.80 1 H NMR (400 MHz, CD3OD) 6 7.7 (mn, 3H), 7.0-7.1 (in, 4H), 3.3 (bin, 4H), 3.1 (bin, 4H), 1.5 9H).
WO 98/00134 PCT/US97/11133 -154- 0 TFA HN- N- N
COOH
21-9 1-(((4-Piperazin-l-yl)phenyl)-2-oxo-isoindolin-5-oxv)acetic acid (21-9) 21-8 (2.6 mmol) was dissolved in EtOAc and cooled to HCI was bubbled through until the solution was saturated. The reaction mixture was then stirred at 0° for 10 min and concentrated to yield a brown solid. Flash chromatography (10/0.8/0.8 yielded 21-9 as a brown solid. This material was further purified by preparative HPLC to yield pure 21-9 (TFA salt) as an off-white solid.
Rf(10/1/1 EtOH/NH40H/H20)=0.72 1H NMR (400 MHz, DMSO- d6) 8 7.8 2H), 7.7 1H), 7.2 IH), 7.1 3H), 4.9 2H), 4.8 2H), 3.2-3.3 (bd, 8H).
WO 98/00134 PTU9/13 PCTIUS97/11133 155 SCHEME 22 22-1 0 H3 N'
H
22-2 1. pyridine N
H
3
CI
22-3 WO 98/00134 WO 9800134PCTIUS97/11 133 156 SCHEME 22 (CONT'D) 22-3 I NaO
H
'OH
22-4
HAN
I HO I/n-butano I
HI.-
H
2
N-
0OC 4
H
9 22-5
'CH
3 WO 98/00134 WO 9800134PCT[US97/11 133 157 SCHEME 22 (CONTD) HCI
H
2
N'
00C 4
H
9 22-5
'CH
3
HN(CH
2
CH
2
CI)
2 0OCAH 22-6
CH
3
HOI.-
211z WO 98/00134 WO 9800134PCTIUS97/11133 158 SCHEME 22 (CONT'D) 22-7 NaOH/EtOH/reflux 22-8
/PYCLU
NO*.I2- 22-9 WO 98/00134 WO 9800134PCT[US97111133 159 SCHEME 22 (CONT'D) 22-9 C H 3 I nBuLi/CNBr 0 N 22-10
H
2 /Pd/C/HOAc 0 22-11 WO 98/00134 WO 9800134PCT[US97111133 160- SCHEME 22 (CONT'D) 22-11
CS
2 00 3 /BrCH 2
CO
2 Et 22 o C 2 Et tBuO y 0 HCI EtOAc 22-13 rN HCI HNJ NaOH 22-14 WO 98/00134 PCT/US97/11133 161 0 0 Jl 22-2 Cl
H
3 C 'N
CH
3
H
1-Chloro-2-(2-methyl-4-acetamidophenyl)ethanone (22-2) A suspension of 3-acetamido toluene (Aldrich) (33.45 g, 0.224 mole) in nitromethane (100 mL) was treated with chloracetylchloride (69.3 mL, 0.739 mole). The solution became homogenous. Aluminum trichloride (92.3 g, 0.694 mol) was added in g portions over 40 minutes. The temperature rose to 50 0 C and the solution became dark brown. The reaction was heated to 80 0 C for 2 hours, then cooled to room temperature and concentrated to give a thick green oil that was added to crushed ice and stirred with a overhead stirrer. The resulting tan solid was collected in a sintered glass funnel and washed with water and dried to give 22-2 as a tan solid.
Rf (40% EtOAc/Hexanes)=0.14.
1H NMR (300 MHz, CDC13) 8 7.66 1H), 7.58 1H), 7.43 1H), 7.28 1H), 4.62 2H), 2.55 3H), 2.2 3H).
O
H
3 C N CH 3 22-3
H
1 -Pyridinium)-2-(2-methyl-4-acetamidophenyl) ethanone, chloride (22-3) A solution of 22-2 (21.8 g, 0.098 mol) in refluxing ethanol (110 mL) was treated with pyridine (28 mL). A salmon-pink percipitate formed and after 2 hours of reflux the mixture was cooled to WO 98/00134 PCT/US97/11133 -162room temperature, filtered and rinsed with to give 22-3 as an off-white solid.
Rf (9:1:1 EtOH/H20tNH40H) 0.12 1 H NMR (400 MHz, D20) 8.66 2H), 7.92 1H), 7.5 1H), 7.33 1H), 2.4 cold (-20 0 C) EtOH and dried 8.5 1H), 8.08 2H), 3H), 2.1 3H).
22-4 2-Methyl-4-amino-benzoic acid 22-4 To a 60°C solution of 22-3 (23.5 g, 0.077 mol) in (155 mL) was added 2.5 M NaOH solution (160 mL). An orange percipitate formed. The reaction was heated to reflux for 2.5 hours, then the homogenous solution was cooled to room temperature and treated with 2.5 M HC1 (160 mL) and stirred overnight. The pH of the solution was adjusted to pH 4-5 with NaOH solution and the yellow percipitate that formed was washed with water and dried to give 22-4 as a gummy yellow solid.
Rf(9:1:1 EtOI-/H20/NH40H)=0.76.
1H NMR (300 MHz, CD30D) 8 7.75 1H), 6.43 2H), 2.42 (s, 3H).
WO 98/00134 PCT/US97/11133 163- 0 HCL* OC 4 9
H
2 N" CH 3 n-Butyl 2-methvl-4-amino-benzoate. hydrochloride (22-5) A suspension of 22-4 (8.8 g, 58 mmol) in n-butanol (200 mL) was cooled to 0°C under argon and saturated with HC1 gas. A salmon-pink percipitate formed. The solution was heated to reflux for 18 hours then evaporated. The residue was dissolved in EtOAc and saturated NaHCO3 and the layers were separated. The aqueous layer was washed repeatedly with EtOAc, the EtOAc layers were combined, dried with brine and MgSO4, filtered and evaporated. The oily residue was dissolved in CHC13 and evaporated to give 22-5 as a brown solid.
Rf (20% EtOAc/Hexanes) 1 H NMR (300 MHz, CDC13) 8 7.83 1H), 6.48 2H), 4.23 2H), 3.9 (bs, 2H), 2.53 3H), 1.7 2H), 1.48 3H), 0.95 3H).
HCI. HN N /O 22-6 bC 4
H
9
CH
3 n-Butvl 2-methvl-4-piperizin-l-yl-benzoate. hydrochloride (22-6) A solution of 22-5 (10.7 g, 52 mmol) in n-butanol (285 mL) was treated with bis chloroethylamine hydrochloride (9.22 g, 52 mmol) and refluxed for one week. The ethanol was removed in vacuo and the residue was diluted with EtOAc. A percipitate formed and was collected and washed with EtOAc and dried to give 22-6 as tan solid.
Rf MeOH/CHCl3 saturated with NH3)=0.31 WO 98/00134 PCT/US97/11133 164- 1H NMR (400 MHz, CD30D) 8 7.88 IH), 6.88 2H), 4.24 (t, 2H), 3.55 4H), 3.36 4H), 2.56 3H), 1.73 2H), 1.5 (m, 2H), 1.0 3H).
tBuO 0 22-7
N
O
N
OC
4
H
9
CH
3 n-Butyl 2-methyl-4-(4-(1,1-dimethylethoxycarbonylpiperizin- l-yl)benzoate acid (22-7) A solution of 22-6 (10.5 g, 33.3 mmol) in CH2C12 (100 mL) was cooled to 0°C and treated with triethylamine (18.8 mL, 0.133 mol) and di-tert-butyldicarbonate (14.75 g, 66 mmol). The solution was allowed to warm to room temperature and after 1 hour was washed with KHSO4, brine, dried over MgSO4, filtered and evaporated to give 22-7 as brown oil.
Rf EtOAc/Hexanes)=0.4 1 H NMR (300 MHz, CDC13) 8 7.9 2H), 6.68 2H), 4.23 2H), 3.55 4H), 3.38 4H), 2.6 3H), 1.75 2H), 1.5 11H), 0.95 3H).
tBuO N N 22-8 0
OH
CH
3 2-Methyl-4-(4-(1,1 -dimethylethoxycarbonylpiperizin-1 -yl) benzoic acid (22-8) A solution of 22-7 (2.55 g, 6.78 mmol) in 4:1 (125 mL) was treated with NaOH (2.7 g, 67.8 mmol) and refluxed overnight. The solvents were removed and the residue was partitioned between EtOAc and 10% KHS04. The layers were separated and the aqeous layer was washed with EtOAc several times. The organic layers WO 98/00134 PCT/US97/11133 165were combined, washed with brine, dried over MgSO4, filtered and evaporated to give 22-8 as a white solid.
IH NMR (400 MHz,CDCl3) 8 8.0 1H), 6.7 2H), 3.6 4H), 3.34 4H), 2.6 3H), 1.5 9H).
f N /O H 22-9 NN CH 3 tBuO N 0 N-(4-Benzyloxyphenyl)-2-methyl-4-(4-(1, 1 -dimethylethoxycarbonyl) piperizin-1 -yl)benzamide (22-9) A solution of 22-8 (2.06 g, 6.43 mmol) in CH2C12 (50 mL) was treated with p-benzyloxy analine hydrochloride (Aldrich, 1.66 g, 7.08 mmol), diisopropyl amine (5.6 mL, 32 mmol) and PYCLU 93.47 g, 9.6 mmol). After stirring for 24 hours the volatile components were removed in vacuo and the residue was dissolved in EtOAc and washed with 10% KHSO4, water, saturated NaHCO3, water and brine, dried over MgSO4, filtered and evaporated. The residue was chromatographed in a gradient of 20 to 50% EtOAc/Hexanes to give 22-9 as an oil.
Rf (50% EtOAc/Hexanes)=0.57 1 H NMR (400 MHz, CDC13) 5 7.4 8H), 7.0 2H), 6.76 2H), 4.08 2H), 3.6 4H), 3.22 (bs, 4H), 2.53 3H), 1.5 9H).
WO 98/00134 PCT/US97/11133 166-
O
0N 0 O 22-10 tBuO N J 0 1 -(4-Benzyloxyphenyl)-5-(4-(1,1 -dimethylethoxycarbonyl)piperizin- 1 vl)isoindolin-2-one (22-10) A solution of 22-9 (0.66 g, 1.31 mmol) in THF (65 mL) was cooled to -78C under argon. n-Butyllithium (2.2M in hexanes, 2.4 mL) was added dropwise to give a deep red solution. The reaction was stirred for 20 minutes, then rapidly transferred via wide bore canulla to a -78 0 C THF solution of cyanogen bromide (2.77 g, 26.2 mmol in mL). The resulting colorless solution was stirred for 25 minutes, then quenched with methanol. The solution was diluted with water, the volatile solvents were removed in vacuo and the remaining water layer was extracted with EtOAc. The EtOAc layer was washed with brine, dried over MgSO4, filtered and evaporated. The residue was chromatographed in a gradient of 20 to 30% EtOAc/Hexanes to give 22-10 as an oil.
Rf (40% EtOAc/Hexanes)=0.25 1 H NMR (400 MHz, CDCl3) 6 7.77 1H), 7.7 2H), 7.44 2H), 7.4 2H), 7.34 IH), 7.0 3H), 6.9 1H), 5.1 2H), 4.73 (s, 2H), 3.6 4H), 3.3 4H), 1.5 9H).
WO 98/00134 WO 9800134PCTIUS97I11133 167 0
N-&O
tBuO YN,) 22-1 0 1 -(4-Hydroxyphenyl)-5-(4-( 1,1 -dimethylethoxycarbonyl)piperizin- 1 yl)isoindolin-2-one (22-11') A solution of 22-10 (0.155 g, 0.31 mmol) in 10 mL glacial acetic acid was treated with 10% Pd/C (0.3 g) and hydrogenated at psi for 5 hours. The solution was decanted from the catalyst, concentrated and the residue chromatographed using a gradient of 60 to 90% EtOAc/Hexanes to give 22-11 as an oil.
EtOAc/lexanes)=O. 19 I H NMR (300 MHz, CDCl3) 8 7.79 lH), 7.55 2H), 7.0 (in, LH), 6.92 (bs, 1H), 6.86 (in, 2H), 4.72 2H), 3.6,(in, 4H), 3.3 (in, 4H), 9H).
WO 98/00134 WO 9800134PCTIUJS97/11133 -168 0 N--0 C0 2 Et 0 Ethyl 1 -dimethylethoxycarbonyl)piperizin- 1 -yl)isoindolin--l1-yl)phenoxyacetate (22-12) A solution of 22-11 (0.045 g, 0.11 nimol) in DMF (5 mL) was treated with cesium carbonate (23 mg, 0.07 mmol) and ethyl bromoacetate (15 uL, 0.0 11 mmol). After 3 hours the solvents were removed in vacuc and the residue was dissolved in EtOAc and washed with water, brine, dried over MgSO4, filtered and evaporated. The residue was chromatographed in a gradient of 30 to EtOAc/Hexanes to give 22-12 as an oil.
EtOAc/Hexanes)=0.36 I H NMR (400 MHz, CDCl3) 8 7.74 (in, 3H), 7.0 (in, 2H), 6.95 (in, 2H), 4.73 2H), 4.62 2H), 4.28 2H), 3.6 (in, 4H), 3.3 (in, 4H), 9H).
WO 98/00134 WO 9800134PCTIUS97/11133 169 0 I o,/0 2 Et HN *-HCI 22-13 Ethyl 4-(2-oxo-5-piperizin- 1 -yl)isoindolin- 1 -yl)phenoxyacetate, hydrochloride (22-13) A solution of 22-12 (0.04 g, 0.08 mmnol) in EtOAc (10 mL) was cooled to -78'C, saturated with HC1 gas, warmed to 0 0 C and evaporated at ambient temperature to give 22-13 as a white solid.
IH NMR (300 MHz, D20) 5 7.54 1H), 7.4 2H), 7.03 (in, 3H), 6.89 2H), 4.6 2H), 4.15 2H), 3.43 (in, 4H), 3.23 (mn, 4H), 1.15 3H).
0 I N O\"C0 2
H
HN,, 22-14 4-(2-Oxo-5-p2iperizin- 1 -l)isoindolin- 1-yI)phenoxvacetic acid (22-14) A solution of 22-13 (0.04 g, 0.08 inmol) in 1:1 (6 m.L) was treated with IN NaOH solution (0.4 mL). The reaction was concentrated and the residue chromatographed in 9: 1:1 EtOHIH2OJNH4OH to give 22-14 as a white solid.
Rf(9: 1:1 EtOH/H2OINH4OH)=0.62 1 H NMR (400 MHz, D20 deutero trifluoroacetic acid) 8 7.49 1H), 7.44 2H), 6.95 (mn, 3H), 6.74 2H), 4.33 (bs, 4H), 3.42 (bs, 4H), 3.25 (bs, 4H).
WO 98/00134 WO 9800134PCT/US97/11133 170 SCHEME 23 jl)OH
H
2 N NO 2 I
BOC
2
O
0 OH23-1 tBuO J NN0
HNO
I CS 2 0 3 /BrCH 2 0 2 Et 0 0 CO2 tBuO N Ja N0 2 H1. H 2 Pd/C 2- 2. CH 3
SO
2
CI
NHSO
2
CH
3
HH
23-3 HCOI /EtOAc23a 0 CO 2
E
HI0H 2 N
NHSO
2
CH
3 23-4 WO 98/00134 WO 9800134PCT[US97/11133 171 SCHEME 23 (CONT'D) 23-4 PCUtBuO 0 0 0 OH tBuO 0 N-N -CHN _Q o 23-5 H
"CO
2 Et
NHSO
2
CH
3 1. HCI/ EtOAc
LOH
jaO,,-CO 2
H
N NHSO 2
CH
3 rN&
H
HNJ 2- WO 98/00134 WO 9800134PCTIUS97/1 1133 -172- SCHEME 23 (CONT'D) tBuO J Nja 0
H
23-2 t~uO 0 H 2 Et 23-7 NO 2 H
-COH
23-8
NO
2 WO 98/00134 WO 9800134PCT[US97/11133 173 0 tBuO
J
N0 2 23-2 1. H 2 Pd/C 2. 3-pyridyISO 2
CI
tBuO
HCI*HI
23-9 23-10 tBuO 0 0
OH
PYCLU
23-11 23-12 rN
HN,,
WO 98/00134 PCT/US97/11133 174- 0"OH tBuO N NO 2
H
23-1 2-Nitro-4-(.1 -dimethvlethoxycarbonvlamino)phenol (23-1) A solution of 2-nitro-4-amino phenol (Aldrich) (20 g, 130 mmol) in THF (500 mL) was cooled to 0 C and treated with ditertbutyldicarbonate (64 g, 293 mmol) and triethylamine (37 mL, 265 mmol). After 24 hours the solution was concentrated and the residue dissolved in EtOAc, washed with 10% KHSO4, saturated NaHCO3, and brine, dried over Na2SO4, filtered and evaporated. The crude bisprotedted material (Rf (40% EtOAc/Hexanes) 0.69) was then dissolved in 400 mL 1:1 THF/H20 and treated with LiOH*H20 (38 g, 1.3 mol).
After stirring at room temperature overnight the solvent was removed and the residue was dissolved in EtOAc and washed with brine, dried with Na2SO4, filtered and concentrated to give 23-1 as a reddish oily solid.
EtOAc/Hexanes)=0.41 1 H NMR (400 MHz, CDC13) 8 10.35 IH), 8.18 1H), 7.58 1H), 7.13 1H), 6.45 (bs, 1H), 1.55 9H).
3, a O C O 2Et tBuO N NOg
H
22 Ethyl 2-(2-nitro-4-(1,1 -dimethylethoxycarbonylamino)phenoxy)acetic acid (23-2) A solution of 23-1 (5 g, 19.7 mmol) in DMF (125 mL) was treated with cesium carbonate (3.17 g, 9.73 mmol), stirred for WO 98/00134 PCT/US97/11133 -175 minutes and treated with ethyl bromoacetate (2.2 mL, 19.8 mmol) at room temperature. After 1.5 hours the solution was concentrated under high vacuum and the-residue was absorbed to silica gel and chromatographed in a gradient of 20 to 30% EtOAc/Hexanes to give 23-2 as a bright yellow solid.
EtOAc/Hexanes)=0.26 1H NMR (400 MHz, CDC13) 8 7.95 1H), 7.5 1H), 6.97 1H), 6.62 (bs, 1H), 4.72 2H), 4.25 2H), 1.5 9H), 1.28 3H).
0 QCOEt HCI 2 E t
H
2 N
NHSO
2
CH
3 23-4 Ethyl 2-(2-methanesulfonamido-4-aminophenoxy)acetic acid hydrochloride (23-4) A solution of 23-2 (2 g, 5.88 mmol) in EtOAc (25 mL) was treated with 10% Pd/C (0.67 and hydrogenated under balloon pressure for 1.5 hours. The solution was filtered through SolkaFloc, and the cake rinsed with EtOAc. The filtrate was not concentrated but was treated directly with methanesulfonyl chloride (3.0 mL, 39 mmol) and pyridine (5.0 mL, 62 mmol) and stirred overnight. The solution was concentrated and the residue was dissolved in EtOAc and washed with 10% KHSO4, saturated Na2CO3, and brine, dried with Na2SO4, filtered and concentrated to give a yellow oil that was chromatographed EtOAc/Hexanes) to give 23-3 (Rf 30% EtOAc/Hexanes) 0.19) contaminated with 23-3a. The mixture (1.7 g) was dissolved in EtOAc (75 mL), cooled to -78 0 C and saturated with HCI gas, warmed to 0 C for 1 hour and concentrated. The residue was partitioned between.
CH2C12 and saturated NaCO3, the layers separated and the aqueous layer extracted with CH2C12. The organic layer was dried over WO 98/00134 WO 9800134PCTTLJS97/11133 -176 Na2SO4, filtered and evaporated and the residue chromatographed EtOAc/Hexanes) to give 23-4 as an off-white solid.
EtOAc/Hexanes) 0.3 1 H NMR (400 MHz, CDCl3) 8 7.7 (bs, IlH), 6.95 I1H), 6.74 IlH), 6.4 1H), 4.6 2H), 4.33 2H), 2.98 3H), 1.3 3H).
tBuO 0 0 N N~ 0 H
CO
2 Et zu- NHSO 2
CH
3 Ethyl 1, 1 -dimethylethoxycarbonyl)piperazin- I -yl)phenylcarbonvlamino)-2-methanesulfonamidophenoxy)acetate (23-5) A suspension of 23-4 (0.125 g, 0.433 mmol) and 1-5 (0.136 g, 0.444 mmol) in CH2Cl2 (4 mL) was treated with diisopropylaxnine (0.3 mL, 1.7 mmol) and PYCLU (0.173 g, 0.48 mmnol) and stirred at room temperature for three days. The solution was concentrated and the residue was absorbed to silica gel and chromatographed in a gradient of 20 to 60% EtOAc/Hexanes to give 23-5 as a pale yell ow oil.
Rf (60% EtOAc/Hexanes)=0.27 1 H NMR (400 MHz, CDCl3) 8 7.82 (dd, ILH), 7.79 ILH), 7.77 I H), 7.54 (bs, 1H), 7.48 1H), 6.90 (in, 3H), 4.7 2H), 4.25 2H), 3.6 (mn, 4H), 3.3 (in, 4H), 3.03 3H), 1.5 9H), 1.3 3H).
WO 98/00134 WO 9800134PCTIUS97/11133 177 o a O,,CO 2
H
N NHSO 2
CH
3 I H r Ne HN 2-(4-(4-(Piperazin- 1 -yl)phenylcarbonylamino)-2-methanesulfonamidophenox y)acetic acid (23-6) A solution of 23-5 (0.093 g, 0. 16 mmol) was dissolved in EtOAc (10 mL), cooled to -78'C and saturated with HC1 gas, warmed to 0 0 C for 1 hour and concentrated. The resulting white solid was dissolved in 1: 1: 1 H20/THF/MeOH, treated with LiOH*H20 (0.03 8 g, 0.9 mnmol) and stirred at room temperature for 1 hour. The reaction was concentrated and chromnatographed (18: 1: 1 EtOH/H2OINH4OH) to give a yellow oil that was diluted with CH2CI2 and evaporated to give 23-6 as white solid.
Rf(9:1 :1 EtOHIH2O/NIH4OH)=-0.48 1 H NMR (400 MI-z, D20 NaOD) 8 7.74 1H), 7.72 1H), 7.21 (s, lH), 7.07 lH), 7.05 1H), 6.9 1H), 6.74 1H), 4.38 2H), 3.15 (in, 4H), 2.85 (mn, 7H).
WO 98/00134 PTU9/13 PCTIUS97/11133 178 tBuO 0 H
\-CO
2 Et 23-7
NO
2 Ethyl 1,1-dimethylethoxycarbonyl)piperazin- 1yl)phenylcarbonvlamino)-2-nitrophenoxy)acetate (23-7) A solution of 23-2 (0.3 g, 0.88 mmol) was dissolved in EtOAc (10 mL), cooled to -78'C and saturated with HC1 gas, warmed to 0 0 C for 1 hour and concentrated to give Ethyl 2-(2-nitro-4arninophenoxy)acetic acid as a white solid that was coupled immediately (0.26 g, 0.88 mmol) to 1-5 (0.29 g, 0.95 mmol) as described for 23-5 to give 23-7 as a yellow solid after chromatography in a gradient of 40 to 100% EtOAc/Hexanes.
EtOAclHexanes)=-0.22
HN
H
-O,
23-8 N 02 2-(4-(4-(Piperazin- 1 -yl)phenylcarbonylarnino) -2-nitrophenoxy)acetic acid (23-8) A solution of 23-7 (0.186 g, 0.352 mmol) in EtOAc was treated first with HCl gas then with LiOH-H20 as described for 23-6 to give 23-8 as a yellow solid after chromatography -in 18:1:1 EtOHIH2OINH-40H).
Rf(1 8:1:1 EtOH/H20/NH4OH)=0.47 IH NMR (400 MHz, D20) 8 8.0 1H), 7.68 (2s, 2H), 7.52 1H), (in, 2H), 3.12 (bs, 4H), 2.85 (bs, 4H).
WO 98/00134 WO 9800134PCTIUS97/11133 179 0 N' 0 H \'-C0 2 Et
NHSO
2 3-pyridyl 23-9 Ethyl 2-(2-(3-pyridyl)sulfonamido-4-( 1,1-dimethylethoxycarbonyl)aminophenoxy)acetate (23-9) A solution of 23-2 (2 g, 5.88 mrnol) in EtOAc (25 mL) was treated with 10% Pd/C and 3-pyridylsulfonyl chloride (JOG, 1989, 54, 389-393) as described for 23-3 to give 23-9 after chromatography in a gradient of 30 to 50% EtOAc/Hexanes as a white solid.
Rf(40% EtOAcfHexanes)=0. 11 1 H NMR (400 MHz, CDC13) 8 9.02 1H), 8.71 1H), 8.1 (in, 2H), 7.4 1H), 7.33 (in, 2H), 6.69 1H), 6.58 IR), 4.4 2H), 4.23 2H), 1.5 9H), 1.25 3H).
tBuO N N -G 0 H ~-0,Et 23-1 1
NHSO
2 3-pyridyl Ethyl 1,1-dimethylethoxycarbonyl)piperazin-1I-yl)phenylcarbonylamino)-2-(3 -pvridylsulfonamido)phenoxy)acetate (23 -11) A solution of 23-9 (0.3 18 g, 0.704 inmol) in EtOAc mL) was treated with HCl gas as described for 23-4 to give 23-10 as a white solid that was coupled directly with 1-5 as described for 23-5 to give 23 -11 -as a oily yellow solid after chromatography in a gradient of to 40% acetone/Hexanes.
EtOAcfHexanes)=0.4 I WO 98/00134 WO 9800134PCT/US97/11 133 -180- 1H NMR (400 MHz, CDCl3) 8
HN
H
-COH
N HS0 2 3-pyridyl 23-12 2-(4-(4-(Piperazin- 1 -yl)phenylcarbonylamino)-2-(2-(3-pyridylsulfonamido)p2henoxy)acetic acid (23-12) A solution of 23-11 (0.047 g, 0.087 mmol) in EtOAc was treated first with HCl gas then with LiOH*H20 as described for 23-6 to give 23-12 as a yellow solid after chromatography in 18: 1:1 EtOH/H20/NH4OH).
Rf(18:1 :1 EtOH/FH20/NH4OH)=0.38 III NMR (400 MHz, D20 NaOD) 858.76 111), 8.5 (in, 111), 8.13 (i,111), 7.7 (in, 2H), 7.45 (in, 1H), 7.12 1H), 7.08 (in, 2H), 6.84 (in 6.67 1 4.13 211), 3.25 (in, 4H), 2.87 (in, 4H).
NONICO
2
E
0 2 N 00CH 3 23-13 Ethyl 2-(2-methoxv-4-nitrophenoxy)acetic acid (23-13) 2-methoxy-4-nitro phenol (Aldrich) (1.0 g, 5.9 inmol) was treated with cesium carbonate and ethyibromoacetate as described for 23-2 to give crude 23-13 after removal of DMF. The crude material was partitioned between water and EtOAc, the organic layer was dried with brine and MgSO4, filtered and evaporated to give 23-13 as a yellow solid.
WO 98/00134 WO 9800134PCTIUS97/1 1133 181 EtOAc/Hexanes)=0.54 IH NMR (400 MHz, CDC13) 8 7.8 1H), 7.76 1H), 6.75 1H), 4.71 2H), 4.2 2H), 3.9 3H), 1.21 3H).
ON0..CO 2 Et
H
2 N a OCH 3 23-14 Ethyl 2-(2-methoxy-4 -aminophenoxy)acetic acid (23-14) A solution of 23-13 (0.7 g, 2.7 rnrnol) in EtOR (10 rnL) was treated with 10% Pd/C (0.14 g) and hydrogenated at balloon pressure. The solution was filtered through Solka-Floc and evaporated to give 23-14 as a tan oil.
IH NMR (400 MHz, CDC13) 8 6.78 1H), 6.33 lH), 6.21 Ifi), 4.59 2H), 4.21 2H), 3.8 3H), 3.45 (bs, 2H), 1.28 3H).
tBuo y N -0NIO 0 H -C0 2 Et 23-15 OCH 3 Ethyl 1,1-dimethylethoxycarbonyl)piperazin- 1yl)phenvlcarbonylamino)-2-methoxyphenoxy)acetate (23-15) Acid 1-5 and amine 23-14 were coupled as described for 23-5 to give 23-15 as brown solid after chromatography in EtOAc/Hexanes.
EtOAc/Hexanes)=0. 13 I H NMR (400 MHz, CDC13) 5 7.8 2H), 7.6 IlH), 6.92 (dl-2H), 6.86 (in, 2H), 4.64 2H), 4.24 2H), 3.9 3.6 (in, 4H), 3.3 (mn, 4H), 1.5 9H), 1.25 3H).
WO 98/00134 WO 9800134PCT[US97/11133 -182- H N N IN -C 02 3 1 6 C0 2
H
OCH
3 2-(4-(4-(4-Piperazin- 1 -yl)phenylcarbonylamino)-2-methoxyphenoxy)acetic acid (23-16) Compound 23-15 was treated with LiOH and HCl gas as described for 23-6 to give 23-16 as a white solid after chromatography in 10:1:1 EtOH/H2OINEI4OH.
Rf (10: 1: 1 EtOHIH2OINH4OH)=O. IH NMR (400 MHz, D20) 5 7.78 2H), 7.15 1H), 7.08 2H), 6.9 (in, 1H), 6.78 1H), 4.4 2H), 3.8 3H), 3.18 (bs, 4H), 2.88 (bs, 4H).
WO 98/00134 WO 9800134PCT[US97/11133 -183 SCHEME 24 HN -24-1
N
H J. Am. Chem. Soc., 1987,109, 3378 1 OC0Br 24-2 tBuO N I 0
H
S1. CH 3 MgBr 2. tert-BuLi 3. 002 0 tBuO N IOH 2-
N
0
H
H
2 N ,CO 2 Et PYCLU/i-Pr 2 NEt HCI CH3 10-4 0 tBuO N N -Q N /ot2- 0 H OH 3 1 HOI/EtOAc 2. LiOH WO 98/00134 PCT/US97/11133 -184- Br 24-2 tBuO N I Y ^N 0
H
2-(1,1 -Dimethylethoxycarbonyl)-7-bromo-1,2,3,4-tetrahydro-9Hpyridor3,4-blindole (24-2) A suspension of 24-1, prepared by the method of Rinehard et al. Am. Chem. Soc., 1987, 109, p 3378-3387) (0.366 g, 1.46 mmol) in CH2C12 (8 mL) was treated with triethylamine (0.61 mL, 4.4 mmol) followed by di-tert-butyldicarbonate (0.38 g 1.7 mmol) for 1 hour at room temperature. The solution was concentrated and the residue chromatographed (20% EtOAc/Hexanes) to give 24-2 as a white solid.
Rf (20% EtOAc/Hexanes)=0.28.
1 H NMR (400 MHz, CDC13) 6 8.0-7.6 1H), 7.46 1H), 7.33 (d, 1H), 7.2 1H), 4.6 (bs, 2H), 3.78 (bs, 2H), 2.76 (bs, 2H), 1.5 9H).
0 24-3 tBuO Na- OH
N
0
H
1-Dimethylethoxycarbonyl)- 1,2,3,4-tetrahydro-9H-pyrido[3,4blindol-7-vl carboxvlic acid (24-3) A solution of 24-2 (0.26 g, 0.734 mmol) in THF (10 mL) was cooled to 0°C and treated with methylmagnesium chloride (3.0 M in THF, 0.29 mL, 0.87 mmol) to give a pale yellow solution. After minutes the solution was cooled to -78 0 C and treated with tBuLi (1.7M in pentane, 4.35 mL, 7.39 mmol) to give a bright yellow solution.
After 10 minutes C02 gas was bubbled vigorously through the solution for 10 minutes. Saturated NH4C1, water and enough 6N NaOH to reach WO 98/00134 WO 9800134PCT/US97/1l 133 185 pH 12 were added and the solution extracted with EtOAc. The EtOAc layer was back extracted with 0.5 NaOH and the aqueous layers combined, acidified to pH 7 and extracted with EtOAc, the EtOAc layer was dried (Na2SO4) filtered and concentrated to give 24-3 as an offwhite solid.
Rf(75:25: 1 CHC13/MeOH/HOAc)=0.48.
I H NMR (400 MHz, DMSO-d6) 8 12.0 bs, 111), 11.2 1H), 7.93 (s, 1H), 7.6 1H), 7.45 1H), 4.6 2H), 3.68 (in, 2H), 2.7 (in, 2H), 1.4 9H).
0 tBuO N N N 0Et 2- YN H OC Et 2oH
OH
3 Ethyl 1,1-dimethylethoxycarbonyl)- 1,2,3,4-tetrahydro-9Hpyrido r3 .4-bi indol-7-yl)carbonylamino)-3 -methylphenoxvacetate (24-4) A solution of 24-3 (0.078 g, 0.25 mmol) and 10-4 (0.303 g, 1.23 mmol) in CH2Cl2 were treated with diisopropylamine and PYCLU as described for 23-5 to give 24-4 as a white solid after chromatography in a gradient of 40 to 60% EtOAc/Hexanes.
Rf (40% EtOAclHexanes)=0. 11 1H NMR (400 MHz, CDCI3) 8 8.5-8.2 (in, 1H), 8.0 11H), 7.75 (d, LH), 7.63 1H1), 7.52 2H1), 6.83 11H), 6.80 1H), 4.7 (bs, 2H), 4.6 2H), 4.28 2H), 3.8 (bs, 2H), 2.83 (bs, 2H), 2.82 3H), 9H), 1.3 3H).
WO 98/00134 WO 9800134PCTIUS97/11133 186- 0 HN N -QO\CO 2 H 24-5 Na H
CH
3 3-Methyl-4-(( 1,2,3,4-tetrahydro-9H-pyrido[3,4-b]indol-7vl'carbonylamino)p2henoxvaceti c acid (24-5) A solution of 24-4 (0.082 g, 0. 16 mnmol) in EtOAc (10 mnL) was treated first with HCI gas, then with LiOH*H20 as- described for 23- 6 to give 24-5 as a white solid after chromatography in 18:1:1 EtOH/IH2OINH4OH.
Rf (18: 1:1 EtOHIH2OINH4OH)=0.48 1H NMR (400 MHz, D20) 5 7.9 1H), 7.54 (in, 2H), 7.13 LH), 6.84 1H), 6.75 1H), 4.40 2H), 3.8 2H), 3.0 (in, 2H), 2.7 (in, 2H), 2.15 3H).
WO 98/00134 WO 9800134PCT/US97/11 133 tBuO
J
187 SCHEME N ~OH ic H
H
3
C
INBS/THF
Br N'07OH2 H
H
3 C Br
ICS
2
CO
3 IBrC H 2 00 2 Et -2 tBuO 5-1 -0 Br tBuON H
\-CO
2 Et
H
3 C Br HCI/EtOAc WO 98/00134 WO 9800134PCTIUS97/11133 188 SCHEME 25 (!QONT'D) Br HCI -H 2 N 0
H
3 0 Br 25-3 t~uO 0 tBiiO f\0 Br -N N- 25-4
~\-CO
2 Et
H
3 0 Br tBuO 0~ 4, Br 25-4 a N -CO 2 Et tBuO 0 HaC Sr WO 98/00134 WO 9800134PCT/US97/11133 189 SCHEME 25 (CONT'D) 25-4 25-4a I LOH t~uO 0 0/ NN&
H
25-5 0 C0 2
H
H
3 C Br C0 2
H
WO 98/00134 WO 9800134PCT[US97/11 133 -190- 0 Br tBuO N /k N OH 25-1 H
H
3 C Br 2,6-Dibromo-3 -methyl-4-( 1,1 -dimethylethoxycarbonyl)aminophenol- (25-1) A solution of 10-2 (1.0 g, 450 mmol) in 20 mL THE under argon was treated with N-Bromosuccinimide (1.6 g, 9 mmol) for 2 hr.
The solution was concentrated and the residue was resuspended in carbontetrachioride and filtered. The filtrate was concentrated and chromatographed (15% EtOAc/Hexanes) to give 25-1 as a white solid.
Rf(20% EtOAc/Hexanes)=O.56 I H NMR (400 MHz, CDCl3) 6 7.79 (bs, I1H), 6.08 (bs, I1H), 5.8 I1H), 2.33 3H), 1.43 9H).
t~ 0- Br H -C0 2 Et
H
3 C Br 25-2 Ethyl 2-(2,6-dibromo-3 -methyl-4-( 1,1-dimethylethoxycarbonyl)aminop~henoxy)acetate (25-2) A solution of 25-1 (0.6 g, 1.57 nirnol) in DMF was treated with cesium carbonate and ethyl bromo acetate as described for 10-3 to give 25-2 as a tan solid EtOAclHexanes)=0.56 1 H NMR (400 MHz, CDGI3) 8 8.0 (bs, ILH), 6.2 1(bs, IlH), 4.56 2H), 4.3 2H), 2.35 3H), 1.5 9H), 1.33 3H).
WO 98/00134 WO 9800134PCT/US97111133 191 Br
H
2 N 0 \-C0 2 Et
H
3 0 Br Ethyl 2-(2.6-dibromo-3 -methyl-4-aminop~henoxy)acetate (25-3) A solution of 25-2 (0.6 g, 1.29 mmol) in EtOAc (10 mL) was treated with HCI gas as described for 16-10 to give 25-3 as a tan solid.
IH NMR (400 MHz, DMSO) 8 7.0 l1H), 4.8-4.4 2H), 4.41 2H), 4.2 2H), 2.18 3H), 1.2 3H).
tBuO 0 Br 0> N-C N 0 25-4
H
3 C Br Ethyl 2-(2,6-dibromo-3 -methyl-4-(4-(N-( 1,1-dimethylethoxycarbony l)pip~erizin-4-yl)p2henvlcarboxami de)p2henoxy) acetate (25-4) .A solution of 25-3 (0.520 g, 1.29 mnmol) and 1-5 (0.395 g, 1.29 mmol) in CH2Cl2 was treated with chloro-NN,N'N',-bis(pentamethylene)formamidinium hexafluorophosphate (0.504 g, 0. 1.4 minol) and diisopropylethyl amnine (0.9 mL, 5.16 mmol) and stirred at room temperature for 24 hours. The solution was diluted with EtOAc and washed with H20, 10% KHSO4, saturated NaHCO3 and brine, dried over MgSO4, filtered and evaporated. The residue was chromatographed (silica gel 30% EtOAcfI-exanes) to give a mixture of 25-4 and 25-4a.
Rf25-4a(50% EtOAclHexanes):=0.45.
WO 98100134 WO 9800134PCT/US97/11133 -192- 1H NMR (400 MHz, CDCl3) 6 8.0 (in, 2H), 7.8 2H), 7.5 I H), 6.93 2H), 6.85 1H), 4.6 2H), 4.3 2H), 3.6 (bs, 8H), 3.35 (in, 8H), 2.4 3H), 1.45 9H), 1.35 3H).
Rf2 -4(0 Et~lexanes)=0.37.
1H NMR (400 MHz, CDCI3) 8 7.6 2H), 6.7 2H), 4.6 2H), 4.3 2H), 3.55 (bs, 4H), 3.3 (bs, 4H), 2.38 3H), 1.45 9H), 1.33 (t, 3H).
tBuO 0B N N-0N0 25-5
\-CO
2
H
H
3 C Br 2- (2,6-Dibromo-3 -methyl-4-(4-(4-( 1 -dimethylethoxycarbonyl)pRiperizin-4-yl)phenylcarbonylamino)phenoxy) acetic acid (25-5) A solution of 25-4 and 25-a. (0.3 g) in 1: 1: 1 was treated with LiOH (0.084 g, 2 minol) at After 1 hour the reaction was diluted with EtOAc and 10% KHS04 and the layers were separated. The organic layer was washed with brine, dried with MgSO4, filtered and evaporated to give 25-5 as a clear oil after chromatography in 9:0.5:0.5 CH2Cl2/N'eOH/HOAc.
Rf(9:0.5 :0.5 CHCI3/MeOHIHOAc)=0.6.
1H NMR (400 MHz, CD3OD) 6 7.90 2H), 7.6 2H), 7.05 2H), 4.55 2H), 3.6 4H), 3.3 (bs, 4H), 2.35 3H), 1.5 9H).
WO 98/00134 WO 9800134PCTIUS97/11 133 193- HN N0 Br HN\- -0 HN-0-0 25-6 HCI
-CO
2
H
H
3 C Br 2-(2,6-Dibromo-3 -methyl-4-(4-piperizin-4-yl)phenylcarbonyl-amino) p2henoxy) acetic acid, hydrochloride (25-6) A slurry of the intermediate acid (0.4 g, 0.6 mimol) in EtOAc was cooled to -78'C and saturated with HCL gas. The reaction was warmed to 0 0 C, then concentrated in vacuc to give 25-6 as the HCl salt.
Rf(1O0:0.5:0.5 EtOH/H20/NH4OH) 0. 18.
1 H NMR (400 MHz, D20) 8 7.73 2H), 7.23 IRH), 7.02 2H), 4.3 2H), 3.1 (bs, 4H), 2.82 (bs, 4H), 2.1 3H).
WO 98/00134 WO 9800134PCTIUS97/1 1133 194- SCHEME 26
H
2 N- ~OH 0 2
N
I BOC2
OTEATHF
H
2 NQ 0J OtBu 26-1 0 2
N
I1. BOC 2
O/DMAP
2. LiOH 0 tBuON N OH 26-2 H
-Q
0 2
N
I
CS
2 CO 3 /BrCH 2 0 2 Et tBuO N 0 O CO 2 Et 26-3 H
-Q
0 2
N
IHCI/EtOAc I26- HCI .H 2 N Q O~ CO 2 Et 2- 0 2
N
WO 98/00134 WO 9800134PCT[US97/11133 195 SCHEME 26 (CONT'D)
PYCLU
tBuO 0 0/ N-o
H
tBuO N \jN
-CO
2 Et Pd/C H 2 WO 98/00134 WO 9800134PCTIUS97/11133 196- SCHEME 26 (CONT'D) tBuO 0 CO 2 Et 26-6 I C'S 2
CAH
26-7
NHSO
2
CAH
1 LIOH 2. HCI/EtOAc HN\- -c p N
C
2
H
H-
NHS0 2
C
6
H
WO 98/00134 PCT/US97/11133 -197- 0 HzN O J OtBu 26-1 0 2
N
1-(1.1 -dimethylethoxycarbonvloxy)-3-nitro-4-aminophenol (26-1) A solution of 3-nitro-4-amino phenol (Aldrich) (8 g, 52.6 mmol) in THF (250 mL) was cooled to 0°C and treated with ditertbutyldicarbonate (24 g, 110 mmol) and triethylamine (14 mL, 105 mmol). The solution was allowed to warm slowly and after 24 hours was concentrated and the residue dissolved in EtOAc, washed with KHSO4, saturated NaHCO3, and brine, dried over Na2SO4, filtered and evaporated to give 26-1 as a brown oil.
1 H NMR (400 MHz, CDC13) 6 7.94 1H), 7.21 (dd, 1H), 6.8 1H), 6.04 (bs, 2H), 1.5 9H).
0 tBuO N OH 26-2
O
2
N
3-Nitro-4-(1,1 -dimethylethoxvcarbonylamino)phenol (26-2) A solution of 26-1 (52.6 mmol) in dichloroethane (150 mL) was added at a rapid drip to a refluxing solution of di-tertbutyldicarbonate (12 g, 52.6 mmol) in dicloroethane (100 mL) and the resulting mixture was refluxed for 20 minutes, then cooled to room temperature and stirred overnight. Triethylamine (52.6 mmol) and 2,6dimethylaminopyridine (1.3 g, 10.5 mmol) were added and the solution refluxed for 2 hours and stirred at room temperature overnight. The solvents were removed and the residue was dissolved in 240 mL 1:1:1 THF/MeOH/H20 and treated with LiOH-H20 (22 g, 526 mmol) for 48 WO 98/00134 PCT/US97/11133 -198hours. The solution was diluted with water, 10% KHSO4 and EtOAc and the layers separated. The organic layer was washed with water and brine, dried over MgSO4, filtered and evaporated. The residue was chromatographed (20% EtOAc/Hexanes) to give 26-2 as a yellow solid.
Rf (20% EtOAc/Hexanes)=0.26 1 H NMR (400 MHz, CDC13) 5 9.5 (bs, 1H), 8.35 1H), 7.61 1H), 7.13 1H), 5.33 1H), 1.55 9H).
0 tBuO N O C0 2 Et 26-3 0 2
N
Ethyl 2-(3-nitro-4-(1,1 -dimethylethoxycarbonylamino)phenoxy) acetate (26-3) A solution of 26-2 (2 g, 7.87 mmol) in DMF (75 mL) treated with cesium carbonate (1.28 g, 3.93 mmol), stirred for minutes and treated with ethyl bromoacetate (0.8 mL, 7.92 mmol) at room temperature. After 1.5 hours the solution was concentrated under high vacuum and the residue was absorbed to silica gel and chromatographed in 10% EtOAc/Hexanes to give 26-3 as a yellow solid.
EtOAc/Hexanes) 0.18 IH NMR (400 MHz, CDC13) 8 9.4 (bs, 1H), 8.44 1H), 7.65 1H), 7.26 1H), 4.63 2H), 4.24 2H), 1.55 9H), 1.3 3H).
WO 98/00134 WO 9800134PCTIUS97/11 133 199
HCI.
H
2 N O CO 2 Et 26-4 0 2
N
Ethyl 2-(3-nitro-4-aminophenoxy)acetate. hydrochloride (26-4) A solution of 26-3(0.5 g, 0.2.5 nunol) was dissolved in EtOAc (15 mL), cooled to -78'C and saturated with HC1 gas, warmed to 0 0 C for 1 hour and concentrated to give 26-4 as an orange solid.
Rf (10% MeOH/CHC13 saturated with NH3)=O. 13 IH NMR (400 MHz, CD3OD) 8 7.51 1H), 7.18 (dd, LH), 7.0 (d, 1H), 4.68 2H), 4.24 2H), 1.15 3H).
tBu O/ 0 0 N 0 111 -CO 2 Et 26- H
-Q
0 2
N
Ethyl 2-(3-nitro-4-(4-(4-( 1,1-dimethylethoxycarbonyl)piperazin- 1vl)p2henyl carbonyl amino)p2henoxy)acetate (26-5) A solution of 26-4 (0.65 g, 2.46 rimol) and 1-5 (1.5 g, 4.9 mmol) were treated with PYCLU and diisopropylamine as described for 23-5 to give 26-5 and 26-5a after chromatography in EtOAc/Hexanes.
EtOAclHexanes)=0. 16 1H NMR 26-5 (400 MHz, CDCl3) 8 8.95 LH), 7.9 2H),-7.75 (s, 1H), 6.95 2H), 4.65 2H), 4.28 2H), 3.6 (in, 4H), 3.35 (in, 4H), 9H), 1.3 3H).
WO 98/00134 PCT/US97/11133 -200tBuO 0 H 26-7
NHSO
2
C
6
H
Ethyl 2-(3-phenylsulfonamido-4-(4-(4-(1,1 -dimethylethoxycarbonyl)piperazin-1 -yl)phenvlcarbonylamino)phenoxy)acetate (26-7) A solution of 26-5 (0.4 g, 0.76 mmol) in EtOH (10 mL) was treated with 10% Pd/C (0.070 and hydrogenated under balloon pressure for 1.5 hours. The solution was filtered through SolkaFloc, and the cake rinsed with EtOAc. The filtrate was concentrated to give 26-6.
Rf(10% MeOH/CHC13 saturated with NH3)=0.72 The crude amine was dissolved in pyridine (3 mL) and treated with phenylsulfonyl chloride (0.10 mL, 0.83 mmol) and stirred for four hours. The solution was diluted with EtOAc and washed with water and brine, dried with MgSO4, filtered and concentrated to give a yellow oil that was chromatographed (gradient 30% EtOAc/Hexanes to 100% EtOAc) to give 26-7 as an oil.
(Rf 30% EtOAc/Hexanes) 0.85) 1 H NMR 26-5(400 MHz, CDC13) 8 8.04 1H), 7.7 2H), 7.65 (d, 2H), 7.5 2H), 7.42 1H), 7.38 2H), 6.9 2H), 6.8 1H), 6.6 1H), 4.46 2H), 4.25 2H), 3.6 4H), 3.3 4H), 1.5 (s, 9H), 1.28 3H).
WO 98/00134 WO 9800134PCTIUS97/11133 -201 0 HN N N 0 -,C0 2
H
H
-Q
NHSO
2 0 6
H
2-(3 -Phenylsulfonamido-4-(4-(4-piperazin-1I-yl)phenylearbonylamino )phenoxy)acetic acid (26-8) A solution of 26-7 (0.18 g, 0.28 mmnol) was treated with LiOH and HCl gas as described for 23-6 to give 26-8 as a white solid after chromatography (10:0.5:0.5 EtOH/NI-40H/H20) and further purification via reverse phase HPLC.
1 H NMR (400 MHz, D20) 5 7.6 (in, 3H), 7.48 2H), 7.23 (mn, IH), 7.18 (in, 2H), 7.1 2H), 6.59 1H), 6.5 4.23 2H), 3.2 (mn, 4H), 2.9 (mn, 4H).
WO 98/00134 WO 9800134PCT/US97/11 133 202 SCHEME 27 ,r-COOH 0
H
3
C
0 ClkKNEt2 (71
CS
2 00 3 DMF, Nal, 60 0
C
HCI _H 3 0 27-2 EtOAc, 0 0
C
0 N Ft 2 0
-HCI
H
3
C
27-3
,'-COOH
H
3
C
0K (27-4)
NHCH
3
CS
2 00 3 DMF, Nal, WO 98/00134 WO 9800134PCTIUS97/11133 -203- SCHEME 27 (CONT'D) 0 0 BOO-N N N 00 H
NHCH
3
H
3 0 Zf 0 HCI (g) EtOAc, 000 0 0 HN N 0 eHOH
H
3 0 0 27-6 ,1 -dimethylethoxycarbonyl)piperazin- 1 -yl)phenylcarbonylamino] -3-methyiphenoxyacetic acid; N,N-diethylglycolamide ester (27-2) A solution of carboxylic acid 10-6 (500 mg, 1.07 mmol), N,N-diethyl chloroacetamide 27-1 (0.154 mL, 1.12 mmol), Nal (5 mg, 0.03 mmol), CS2CO3, (173 mg, 0.535 mmol) and DMF (5 mnL) was heated at 60'C for 20 h. The reaction mixture was diluted with EtOAc and washed with water, 10% aqueous KHS04 and brine. Drying (MgSO4), filtration, removal of the solvent in vacuo, and chromatography on silica gel using a gradient of 50 to 90% EtOAc in hexane as eluant gave 1,1-dimethylethoxycarbonyl)piperazin- 1yl)phenylcarbonylamino] -3 -methyiphenoxyacetic acid N,N-diethylglycolarnide ester (21J-2 as a white solid. 1H NMR (CD3OD): 8 1.23 (t, J=7Hz, 1.48 9H), 2.25 3H), 3.32 (in, 8H), 3.58 (in, 2H), 4.83 2H), 4.92 2H), 6.83 (dd, J=3Hz, 9Hz, 1H), 6.90 7.03 (d, J=9Hz, 2H), 7.19 J=(Hz, 7.78 J=9Hz, 2H).
WO 98/00134 WO 9800134PCT(US97/11 133 -204- 4- [4-(Piperazin- I -yl)phenylcarbonylamino] -3-methyiphenoxyacetic acid, N.N-diethylglycolamide ester, hydrochloride _27-3) To a stirred solution of EtOAc (7 mnL) saturated with HCI gas was added ester 27-2 (500 mg, 0.86 nimol), suspended in EtOAc (3 mL). The mixture was stirred for 1 h at ambient temperature, purged with dry argon for 30 min and concentrated in vacuc to give 4-[4- (piperazin- 1-yl)phenylcarbonylamino] -3-methyiphenoxyacetic acid N ,Ndiethyiglycolamide ester, hydrochloride (27-3) as a white solid. 1H NMR (D20): 5 1.11 J=7Hz, 3H), 1.21 J=7Hz, 3H), 2.23 311), 3.37 J=7Hz, 4H), 3.44 (in, 4H), 3.62 (in, 4H), 4.96 2H), 5.01 (s, 2H), 6.92 J=9Hz, 1H), 7.00 1H), 7.21 (mn, 3H), 7.90 J=9Hz, 2H).
1, 1 -Dimethylethoxycarbonyl)piperazin- 1 -yl)phenylcarbonytamnino] -3 -iethylphenoxyacetic- acid: N-methyl g ivolamide -ester (27-5) A solution of carboxylic acid 10-6 (500 mg, 1.07 inmol), N-methyl chloroacetamide 27-4 (0.121 mg, 1.12 mmol), Nal (5 mg, 0.03 mmol), CS2CO3, (173 mg, 0.535 mmol) and DMF (5 mL) was heated at 60'C for 20 h. The reaction mixture was diluted with EtOAc and washed with water, 10% aqueous KHS04 and brine. Drying (MgSO4), filtration, removal of the solvent in vacuo, and chromatography on silica gel using EtOAc as eluant gave 1,1 dimethylethoxycarbonyl)piperazin- 1 -yl)phenylcarbonylamino] -3methylphenoxyacetic acid N-methylglycolamide ester as a white solid. I 1 H NMR (CD3OD): 8 1.49 9H), 2.31 3H), 2.77 3H), 3.29 (mn, 4H), 3.61 (in, 4H), 4.67 211), 4.75 2H), 5.85 (br s, 111), 6.87 (in, 2H), 6.95 J=(Hz, 2H), 7.47 11H), 7.78 (in, 1H1), 7.79 J=9Hz, 2H).
4- [4-(Piperazin- 1 -yl)phenylcarbonylamino] 3-methylphenoxyacetic acid: N-methyl glycolamide ester. hydrochloride (27-6) HCl gas was rapidly bubbled through a solution of 1 -dimethylethoxycarbonyl)piperazin- I -yl)phenylcarbonylamino]-3methylphenoxyacetic acid N-methylglycolamide ester (27-5) (325 mg, WO 98100134 PCTIUS97/11133 -205- 0.60 nimol) in EtOAc (30 mL) at 0 0 C for 20 min. The mixture was aged 40 min., purged wityh argon and concentrated in vacuo. The solid was triturated with EfOAc and Et2O to give 4-[4-(piperazin- 1 yl)phenylcarbonylamino] -3-methyiphenoxyacetic acid Nmethyiglycolamide ester, hydrochloride (27-6) as a white solid. IH NMR (D20): 8 2.23 3H), 2.78 3H), 3.43 (in, 4H), 3.62 (in, 4H), 4.75 2H), 4.95 2H), 6.90 J=8Hz, 1H), 6.99 1H), 7.21 (mn, 3H), 7.90 J=9Hz, 2H).
WO 98/00134 WO 9800134PCT/US97/11 133 206 SCHEME 28 1. HO 3 S /N 2 +Cl 2. Na 2
S
2
O
4
OH
NOCH
3
NH
2 Boc 2 O, CHC1 3
OH
NOCH
3 NHBoc Br COQEt
CS
2 C0 3
DMF
ref lux 28-3 O COQEt
N'OCH
3 NHBoc HCI (gas) EtOAc, 000 0 COOEt
*HCI
NOCH
3
NH
2 28-4 28-4 28-5 WO 98/00134 WO 9809134PCTIUS97/11133 207 SCHEME 28 (CONT'D) Boo-N NO COQH CI P F 6 No i-Pr 2 NEt, CH 2
CI
2 LiOH DME,
H
2 0 HOI (gas) EtOAc, 0 0
C
COO H
-HCI
,r-COOH 0
OCH
3 WO 98/00134 WO 9800134PCTIUS97/11133 -208- 4- [4-(lI -Piperazinyl)phenylcarbonylamino] -3 -methoxyphenoxyacetic acid, hydrochloride (28-8) Using a. sequence essentially the same as described for compound 159 but starting with 3-methoxyphenol piperazinyl)phenylcarbonylamino] -3 -methoxyphenoxyacetic acid, hydrochloride as prepared. IH NMR (D20): 8 3.33 J=3.9Hz, 4H), 3.51 J=3.9Hz, 4H), 3.75 3H1), 4.62 2H), 6.51 J=8.6Hz, lH), 6.64 111), 7.05 J=7.4Hz, 2H), 7.32 J=8.6Hz, 1H), 7.75 (d, J= 7.4Hz, 2H).
WO 98/00134 WO 9800134PCT/US97/11133 209 SCHEME 29 o OH 3 C H 3
OH
12, pyridine, 10000 29-1 BOON Na NH 2 i-Pr 2 NEt, CH 2 C1 2 BOON
N-&
29-3
H
3
C
BrCH 2 000Bu t 0S 2 00 3
DMF
WO 98/00134 WO 9800134PCT/US97/11133 -210- SCHEME 29 (CONT'D)
-H
BOCN -N /-.COOBUI 29-4
H
3
C
HCI (gas) EtOAc, 000
H
HN pCOOH 0 4-Hydroxy-2-methylbenzoic acid (29-2) Following the procedure described in US 92-852870 920317, 4-hydroxy-2-methylbenzoic acid was prepared from 4hydroxy-2-methylacetophenone 29-1. IH NMR (CD3OD): 8 2.52 (s, 3H), 6.64 (in, 2H), 7.84 J=8.4 Hz, 1H).
1-Piperazinyl)phenylaminocarbonyl] -3 -methyiphenoxyacetic acid, hydrochloride (29-5) Using a sequence essentially the same as described for compound 14-6, but starting with 4-hydroxy-3-methylbenzoic acid 1-piperazinyl)phenylaminocarbonyl]-3methyiphenoxyacetic acid, hydrochloride (29-5) was prepared. IH NMR (DMSO-D6): 8 2.36 3H), 3,26 J=7.3Hz, 8H), 3.80 (br s, 4H), 4.73, 2H), 6.81 (dd, J= 2.4, 8.4 Hz, 2H), 6.84 J=2.4Hz, 1H), 6.97 J=8.7Hz, 2H), 7.39 J=8.4 Hz, I1H), 7.62 J=8.7Hz, 2H), 8.72 (br s, I1H), 9.99 ILH).
WO 98/00134 WO 9800134PCTIUS97/11133 -211- SCHEME
OK-
C F 3 1HO 3 S /N 2 4 Cr
OH
2. Na 2
S
2
O
4 Boc 2 O, CHC1 3 ref lux
NH
2 30-2
OH
CF
3 NHBoc Br COOEt KN (TMS) 2 18-C ROWN-6
THF
WO 98/00134 WO 9800134PCTIUS97/11133 -212- SCHEME 30 (CONT'D) COOEt HOI (gas) EtOAc, 0 0
C
*HCI
'CF
3 NHBoc 30-5 Boo-N \/COOH i-Pr 2 NEt, CH 2
CI
2 WO 98/00134 WO 9800134PCT/US97/11 133 -213- SCHEME 30 (CONT'D) Boo- /-COQEt LiOH DME,
H
2 0
CF
3 N N-N /-COOH
CF
3 HOI (gas) EtOAc, 000
*HCI
HN -COOH
CF
3 1.1-Dimethylethoxycarbonyl)amino-3 -trifluoromethyiphenoI (30-3) Using a method similar to that described for- compound 15-3, starting with 3 -trifluorophenol, 1,1-dimnethylethoxycarbonyl)amino -3 -trifluoromethylphenol (2Q-3) was prepared. 1H NMR (CDCl3): 8 1.55 9H), 4.12 (in, 2H), 6.72 1H), 7.12 (in, 1H), 7.26 (in, I1H).
WO 98/00134 PCT/US97/11133 -214- Ethyl 4-(1,1-dimethylethoxycarbonyl amino-3-trifluoromethylphenoxvacetate (30-5) To a 200 mL round bottomed flask with a stirring bar and an argon inlet was added 4-(1,1-dimethylethoxycarbonyl)amino-3trifluorophenol (4.80 g, 17.3 mmol), 18-crown-6 (4.63 g, 17.5 mmol), and THF (69 mL). This mixture was cooled to 0°C in an ice-water bath, then a solution of potassium hexamethyl disilylazide in toluene (34.7 mL, 17.4 mmol) was added followed by ethyl bromoacetate (1.93 mL, 17.4 mmol). This mixture was stirred under argon for 16 h, allowing the cooling bath to expire. This mixture was transferred to a separatory funnel, acidified with 10% aqueous citric acid and extracted with EtOAc. The organic layer was washed with water and brine, dried (MgSO4), filyered and concentrated in vacuo. The BOC group was removed as described for 29-9. This material was chromatographed on silica gel to give ethyl 4-(1,1-dimethylethoxy-carbonyl amino-3trifluoromethylphenoxyacetate (30-5) as an oil. 1H NMR (CDC13): 1.58 3H), 3.93 (br s, 2H), 4.27 2H), 4.56 2H), 6.69 1H), 6.95 (dd, 1H), 7.00 1H).
4-[4-(1-Piperazinyl)phenylcarbonylamino]-3-trifluoromethylphenoxyacetic acid (30-8) Using a sequence similar to that described for compound 12-8 but starting with ethyl 4-(1,1-dimethylethoxycarbonyl amino-3trifluoromethylphenoxyacetate 4-[4-(1-piperazinyl)phenylcarbonylamino]-3-trifluoromethylphenoxyacetic acid 308) was prepared. 1 H NMR (DMSO-d6): 5 3.25 4H), 3.50 4H), 4.84 (s, 2H), 7.09 J=8.0 Hz, 2H), 7.25 2H), 7.39 J=8.4 Hz, 2H), 7.88 J= 8.0 Hz, 2H), 8.82 (br s, 1H), 9.72 1H).
WO 98/00134 PTU9/13 PCTIUS97/11133 -215- SCHEME 31
H
2 N -Q
K..CO
2 Et *HCI HCH N -(NH 10-4
O
2 N& ~O yCI 2C 31-2 0 pyridine, CHq01 2 60020, NaQH 0 EtOH/H 2 0 0 2 NH 2 Et
H
3 0 oNN 31-1 31-3 IEt 3
N,CH
2 01 2 WO 98/00134 WO 9800134PCTILJS97/1 1133 -216- SCHEME 31 (CONT'D) BooN~ N 0 OCO 2 Et 0H 3
C
31-4 I LiH, THF/H 2 0/MeOH -T -0-1-CO 2
H
0
H
3
C
31-5 IO (gas) EtOAc, 00 HNQ- N' 1 T \/HO *HCI 0 H. C WO 98/00134 PCT/US97/11133 -217- Ethyl 4-(4-nitrophenylcarbamate)-3-methylphenoxyacetate (31-1) A solution of ethyl 4-amino-3-methylphenoxyacetate hydrochloride (10-4) (300 mg, 1.22 mmol) in CH2C12 (30 mL) was placed in a 50 mL flask. To this solution 4-nitrophenyl chloroformate (246 mg, 1.22 mmol) and pyridine (0.22 mL) were added at room temperature. The reaction was stirred at ambient temperature 18 h, then concentrated in vacuo to yield ethyl 4-(4-nitrophenylcarbamate)-3methylphenoxyacetate 1 H NMR (CDC13): 8 1.30 j=7.0 Hz, 3H), 2.34 3H), 4.28 j=7.0 Hz, 2H), 4.59 2H), 6.70 (dd, j=8.3, 2.0 Hz, 1H), 6.7 j=2.0 Hz, 1H), 7.40 j=8.3 Hz, 1H), 8.05 (d, Hz, 1H), 8.28 j=7.0 Hz, 2H).
4-(1,1-Dimethvlethoxycarbonyl)-4.4'-bipiperidine (31-3) A solution of bipiperidine dihydrochloride (Aldrich 31-2) (25 g, 0.1 mol) in H20 (100 mL) was placed in a 2L round bottomed flask. A solution of 6N NaOH was added dropwise to adjust the pH to 8-9. The solution was diluted with EtOH (1200 mL) and stirred vigorously at ambient temperature. The resulting mixture was treated with a solution of di-t-butyl dicarbonate (24 g, 0.11 mol) in EtOH (800 mL). Periodically, 6N NaOH was added to maintain pH 8-9. After stirring at ambient temperature 18 h, the reaction was concentrated in vacuo. The residue was dissolved in 1:1 ether:H20 (1000 mL) and the pH adjusted to 12 with 6N NaOH. The aqueous phase was separated and extracted again with ether. The ether phases were combined and washed with brine, 10% citric acid, and H20. The citric acid and H20 washes were combined and the pH adjusted to 12-13 with 6N NaOH, then extracted in 3 portions with ether (600 mL). The ether extract was washed (brine), dried (Na2SO4) and concentrated in vacuo to a clear oil which solidified on standing to yield 4-(1,1-dimethylethoxycarbonyl)- 4,4'-bipiperidine 1H NMR (CDC13): 6 1.25 (br m, 7H), 1.45 9H), 1.66 (br d, 4H), 2.60 (dd, 4H), 3.1(d, j=12 Hz, 2H), 4.21(br s, 2H).
WO 98/00134 WO 9800134PCT/US97/11 133 218 Ethyl-4- 1,1 -dimethylethoxycarbonyl)-4 ,4'-bipiperidinyl- 1carbonyl)aminol -3 -methyliphenoxyacetate (31 -4 A solution of ethyl 4-(4-nitrophenylcarbamate)-3methyiphenoxyacetate (457 mg, 1.22 mmol), 1, 1 -dimethylethoxycarbonyl)-4,4'-bipiperidine (31-3) 327 mg, 1.22 inmol), and triethylamine (0.34 mL, 2.44 nunol) in CH2Cl2 (20 The solution was refluxed 4 h, cooled and diluted with CH2Cl2 (10 mL). The reaction was extracted with H20 (15 mL). The CH2Cl2 extract was washed NaOH, H20, 10% KHSO4, brine), dried (Na2SO4), filtered, then concentrated in vacuc to an oil. The oil was chromatographed on silica using 50% EtOAc/hexane to obtain ethyl-4- (1,1-dimethylethoxycarbonyl)-4,4'-bipiperidinyl-l1-carbonyl)amino] 3-methyiphenoxyacetate (31-4) 1 H NMR (CDCl3): 8 1.27 (br m, 7H), 1.30 j= 7.0 Hz, 3H), 1.46 9H), 1.68 j=-12 Hz, 2H), 1.75 j=l10 Hz, 2H), 2.22 3H), 2.65 j= 12 Hz, 2H), 2.83 j= 12 Hz, 2H), 4.08 j=12 Hz, 1H), 4.25 (br s, 2H), 4.27 j=7 Hz, 2H), 4.58 (s, 2H), 5.97 1H), 6.70 (dd, j=8.3, 2 Hz, 111), 6.73 j=-2 Hz, lH), 7.40 j=8.3 Hz, I H), 1,1 -Dimethylethoxycarbonyl)-4,4'-bipiperidinyl-l1-carbonyl)amino]-3-methylp~henoxyacetic acid (31-5) In a 25 mL round bottomed flask a solution of ethyl-4-[(4- 1 -dimethylethoxycarbonyl)-4,4'-bipiperidinyl- 1 -carbonyl)amino] -3methyiphenoxyacetate (3 1-4) (100 mg, 0.2 mrnol), IM LiOH, (0.6 mL, 0.6 mmol), H20 (5 mL), THF (5 mL), and MeOH (5 mL) was stirred at ambient temperature 3 h. The reaction was concentrated in vacuc, then dissolved in CHC13 (20 mL). The CHC13 extract was washed (IN HCl, and brine), dried (NaSO4), filtered, and concentrated to yield 4- 1,1 -dimnethylethoxycarbonyl)-4,4'-bipiperidinyl- 1 -carbonyl)amino] 3-methylphenoxyacetic acid (3 1H NMR (CDCI3): 8 1.22 (br m, 6H), .1.46 9H), 1.6 (br m, 4H), 2.2 3H), 2.6 (br t, 2H), 2.79 (t, j=lI1 Hz, 2H), 4.0 (br m, 5H), 4.58 2H), 6.1 (dd, j=9, 2 Hz, I1H), 6.75 j =2 Hz, IlH), 7.21 j=9Hz, IlH), 8.1 I1H).
WO 98/00134 WO 9800134PCTIUS97/111133 219 4- -Bipiperidinyl- 1 -carbonyl)amino] -3 -methyiphenoxyacetic acid, hydrochloride (31-6) In a 35 mnL round bottomed flask 4-[(4-(1,1-dimethylethoxycarbonyl)-4,4'-bipiperidinyl- 1-carbonyl)amino] -3 -methyiphenoxyacetic acid (31-5) (83 mg, 0. 17 inmol) was dissolved in CH2Cl2 (00 mL) and cooled to Anhydrous HC1 was bubbled through the solution for 5 min. The reaction was stirred at 0' 4 h, then concentrated in vacuc to yield 4- -bipiperidinyl-l1-carbonyl)amino] -3methyiphenoxyacetic acid, hydrochloride IH NMR (DMSO-d6): 8 1.04 (br d, j=11I Hz, 3H), 1.3 (br s, 3H), 1.62 j=12 Hz, 2H), 1.79 j=1 1 Hz, 2H), 2.07 3 2.67 j- 12 Hz, 2H), 2.74 (br m, 2H), 4.07 j= 12 Hz, 2H), 4.59 2H), 6.62 (dd, j=9, 2 Hz, I1H), 6.70 (d, j=2 Hz, 1H), 6.95 j=9 Hz, 1H), 7.87 lH), 8.2 (br s, 1H), 8.5 (br s, 1H).
WO 98/00134 WO 9800134PCTIUS97/11133 -220- SCHEME 32
CI*PF
6
HO
2
C(CH
2 r->-OH BocN NH H i-Pr 2 NEt, DMF 31-3 32-l a (n=O) 32-lb (n=l' Bocla N )f(CH 2 )n /OH 0 a2-2a (n=O) 32-2b (n=1) IBrC H 2 00 2 t-Bu
CS
2
CO
2
DMF
BoNN (CH 2 )n ~OtB 0 32-3a (n=O) ITFA, CH 2
CI
2 0.
HN N (CH 2 )n /G C2 *TFA 0 32-4a (n=-W WO 98/00134 WO 9800134PCTIUS97/11 133 221 4-(2-(4-Dimethoxyethoxycarbonyl)-(4,4')-bipiperidinyl- I1carbonvl~phenol -(32-2a) In a 50 mnL round bottomed flask a solution of 1,1 dimethylethoxcarbonyl)-4,4'-bipiperidine (31 (400 mg, 1.49 mm), 4hydroxybenzoic acid (206 mg, 1.49 mm), (32-1a), chloro-N,N,N',N'bis(pentamethyl)formamidinium hexafluorophosphate (537 mg, 1.49 mm) and *diisopropylethylamine (0.52 mL, 2.98 mm) in 25 mL DMF was stirred at ambient temperature 48 h. The reaction was concentrated in vacuc, then partitioned between EtOAc and H20. The EtOAc extract was washed (H20, sat'd NaHCO3, H20, 10% KHS04 and brine), dried (Na2SO4), filtered and concentrated in vacuo. The resulting oil was chromatographed on silica using 5% MeOH/CHCJ3 as eluent to yield 4- (2-(4-dimethoxyethoxycarbonyl)-(4,4')-bipiperidinyl)- 1 -carbonyl)phenol (32-2a). 1H NMR (CDCl3): 8 1.2 (br m, 6H), 1.4 9H), 1.58 (br m, 4H1), 4.1 (in, 2H), 4.3 (in, IH), 6.83 j=8.5 Hz, 2H), 7.31 (d, Hz, 2H).
4-(2-(4-Dimethoxyethoxycarbonyl)-(4,4')-bipiperidinyl- 1 -yl-2-oxoethyDb2henol (32-2b) In a manner similar to the preparation of compound 32-2a, 4-(2-(4-dimethoxyethoxycarbonyl)-(4,4')-bipiperidinyl- 1 -yl-2-oxoethyl)-phenol (32-2b) was prepared. 1 H NMR (CDCl3): 5 0.89 (mn, 1 1. 1 (mn, 1.45 9H), 1.5 (br mn, 6H) 2.49 j= 12 Hz, I1H), 2.62 (in, 2H), 2.91 j= 12 Hz, I1H), 3.64 2H). 3.90 (dj-_12 Hz, I1H), 4.1 (mn, 2H), 4.68 j= 12 Hz, I1H), 6.12 (br mn, IlH), 6.77 j=8 Hz, 2H), 7.07 j=8 Hz, 2H).
t-Butyl-4-(2-(4-Dimethoxyethoxycarbonyl)-(4,4')-bipiperidinyl- I carbonyl~phenoxyacetate (32-3a) In a 50 mL round bottomed flask a mixture of dimethoxyethoxycarbonyl)-(4,4')-bipiperidinyl-lI-carbonyl)-phenol (32- 2-a) (378 ing, 0.97 mim), t-butyl bromoacetate (0.16 mL, 1.067 mmn) and cesium carbonate (348 mng, 1.067 mim) in 50 mL DMF was stirred at ambient temperature 18 h. The reaction was concentrated in vacuo, WO 98/00134 WO 9800134PCTIUS97/11 133 -222then partitioned between CHC13 and H20. The CHC13 extract was washed (10% KHSO4, H20, NaHCO3 and brine), dried (Na2SO4), filtered and concentrated in vacuo to yield t-butyl-4-(2-(4-dimethoxyethoxycarbonyl)-(4 ,4')-bipiperidinyl-lI-carbonyl)-phenoxyacetate (22- 3aj. IH NMR (CDCI3): 5 1.2 (in, 6H), 1.42 9H), 1.45 9H), 2.68 (in, 4H), 2.61 2H), 2.8 (in, 2H), 4.18 (in, 4H), 4.5 2H), 6.9 j=8 Hz, 2H), 7.35 j=8 Hz, 2H).
t-Butyl-4-(2-(4-dimethoxyethoxycarbonyl)-(4 ,4')-bipiperidinyl- I-yl- 2-oxo-ethvl)p2henoxvacetate (32-3b) In a manner similar to the preparation of 32-3a, t-butyl-4- (2-(4-dimethoxyethoxycarbonyl)-(4,4')-bipiperidinyl- 1 -yl-2-oxo-ethyl)phenoxyacetate (32-3b) was prepared. I H NMR (CDCl3): 8 0.88 (in, I1H), 1. 11 (in, 5H), 1.45 9H) 1.48 9H), 1.61 (in, 4H), 2.48 (t, j=l12Hz, I1H), 2.61 (br t, 2H), 2.8 8 j= 13 Hz, I 3.66 2H), 3.88 j= 13 Hz, I1H) 4.1 (in, 2H), 4.49 2H), 4.68 j= 13 Hz, 1 6.84 j=8.8 Hz, 2H), 7.15 j=8.8 Hz, 2H).
L4 -(r44'-1 -B ipiperidinyl -1 -carbonyl)p2henoxyl acetic acid (32-4a) In a round bottomed flask, t-butyl-4-(2-(4-dimethoxyethoxycarbonyl)-(4,4')-bipiperidinyl-l1-carbonyl)-phenoxyacetate (12-3a) (56 mng, 0.11 mm) was stirred with trifluoracetic acid (2.5 mL) and CH2C12 (5 m.L) at 0' 18 h. The reaction was concentrated in vacuo to yield -bipiperidinyl- I -carbonyl)phenoxy] -acetic acid 4a). I H NMR (DMSO-d6) 8 1.11 (mn, 2H), 1.32 (mn, 4H), 1.78 (mn, 2H), 1.80 j=12 Hz, 2H), 2.81 (dd, j=1I 1 Hz, 4H), 3.28 j=1 1 Hz, 2H), 4.73 2H), 6.94 j=8.5 Hz, 2H), 7.32, J=8.5, 2H), 8.16 (br s, 1H), 8.47 (br s, 1H).
-Bipiperidinyl- Il-yl -2-oxo-ethyl)phenoxy] acetic acid (32-4b) In a manner similar to the preparation of (32-4a), 4-(2- -bipiperidinyl- 1 -yl-2-oxo-ethyl)-phenoxy] -acetic acid (32-4b) was prepared. I 1 H NMR (CDCI3): 5 0.81 (in, 1H), 1.07 (mn, 1H), 1.36 (br s, WO 98/00134 PCTIUS97/11133 -223- 2H), 1.62 j=13 Hz, 1H), 1.74 j=12 Hz, IH), 1.87 (br s, 2H), 2.56 j1l 1 Hz, IH), 2.94 (in, 3H), 3.36 j=12 Hz, 2H), 3.70 (in, 3H), j= 13 Hz, I 4.57 j= 13 Hz, I1H), 4.64 2H), 6.89 j=8.5 Hz, 2H), 7.17 j=8.5 Hz, 2H).
WO 98100134 WO 9800134PCTIUS97/1 1133 -224- SCHEME 33 NMM, H N
NMP
1100
CO
2 Et NQD/- NQo- OO 2 Et 33-1 N/-N--CO 2
H
33-2 PYCLU, iPr 2 NEt,
DMF
H
2 N-Q l. 0\/-R
H:
3 C 10-4 WO 98/00134 WO 98/0013 PCTIUS97/1 1133 -225- SCHEME 33 (CONT'D) ND, 0 N 0 OCOEt
H
3
C
33-3
KOH,
TH F/H 2 0 H- 0
CO
2
H
H
3 0 N\/-ND
-C
2 Et 33-1 Ethyl 4-pvridylpiperidin-4-vlcarboxylate (33-1) Ethyl isonipecotate (6.0 g 38.66 mmol), 4-chioropyridine hydrochloride (5.9 g, 38.66 mmol) and N-methyhnorpholine (9.3 g mL, 85.00 mmol), were dissolved in N-methylpyrrolidine (50 mE) and the resulting solution was heated at 100TC for 48 h. The solution was concentrated in vacuc and the residue was dissolved in EtOAc and washed with water and brine (2 x 100 then dried (Na2S 04) and evaporated. The resulting residue was purified by flash chromatography MeOH/CHC13) to afford 33-1 as a crystalline solid. IH NMR (CDCI3) 6 8.21 J=6.8 Hz, 2H), 6.78 J=6.8 Hz, 2H), 4.18 J=7.0 Hz, 2H4), 3.85 (in, 2H), 3.10 (in, 2H), 2.61 (in, I1H), 2.05 (in, 2H), 1.85 (mn, 2H), 1.23 J=7.0 Hz, 3H-).
WO 98/00134 WO 9800134PCTIUS97/11133 -226- 33-2 4-Pyridvlpip~eridin-4-ylcarboxylic acid (33-2) A solution of 33-1 (10 g, 42.7 mmol) in THF (50 ml-) was treated with IN LiOH (47 mL, 47.0 mimol) and water (50 mL). The resulting solution was stirred at ambient temperature for 12 h. The solution was concentrated and the aqueous residue was cooled to 0 0
C,
then adjusted to pH 6 with IN HCI. The resulting solid was collected by filtration and dried in vacuo to afford 33-2 as a white solid. 'fl NMR (D20) 8 7.95 J=6.8 Hz, 2H0, 6.73 J=6.8 Hz, 2h), 3.76 (d, J=12.8 Hz, 2H0, 2.81 (in, 2H), 2.20 (in, lH), 1.85 J=12.9 Hz, 2H), 1.55 (mn, 2H).
Dx ND N5 0
CO
2 Et
H
3
C
Ethyl 4-(Pyridyl)(piperidin-4-yl)-carbonylamino-3 -methylphenoxyacetate (33-3) Ethyl 4-amino-3-methylphenoxyacetate hydrochloride (IQA4) (0.35 g, 1.41 mmol), 4-(Pyridyl)(piperidin-4-yl)-carboxylic acid (0.30 g, 1.41 mmol), chloro-N, N, N'-bis(pentamethylene)formnamidinium hexafluorophosphate (0.50 g, 1.41 mnmol), and diisopropylethylainine (0.25 mL, 1.41 mmol) were dissolved in dimethylformamide (15 mL). The solution was stirred at ambient temperature for 48 h and the solvent removed in vacuo to give an oil.
This material was chromnatographed on silica gel using 5:95 methanolammonia saturated chloroform as eluant to afford 33-3 as a beige solid.
1 H NMR (CDC13): 6 8.27 j=5.6 Hz, 2H), 7.5 3 j=8.8 Hz, I H), 7.03 1H), 6.78 1H), 6.73 j=8.8 Hz, I H) k, 6.68 j=6.1 Hz, 2H), 4.59 2H), 4.27 j=7.1 Hz, 211), 3.97 j= 13.2 Hz, 2H), 2.95 WO 98/00134 WO 9800134PCT/US97/11133 -227j=1 1.7 Hz, 2H), 2.53 (in, 1H), 2.06 j=10.7 Hz, 2H), 1.92 (in, 2H), 1.73 3H), 1.30 j=7.1 Hz, 3H).
ND HN--5 0
,CO
2
H
H
3
C
33-4 4-(Pyridyl)(piperidin-4-yl)-carbonylamino-3-methylphenoxyacetate (33-4) A solution of Ethyl 4-(Pyridyl)(piperidin-4-yl)-carbonylamino-3-methylphenoxyacetate (33-3) (0.05 g, 0.126 inmol) in tetrahydrofuran (10 m-fL) was treated with I N lithium hydroxide 13 mL, 0.132 minol) and H20 (10 mL). The resulting solution was stirred at ambient temperature and the solvent removed in vacuc to give 33-4 as a colorless glass. IH NMR (CD3OD) 8 8.09 j=6.6 Hz, 2H), 7.09 (d, j=8.6 Hz, 2H), 6.87 6.8 Hz, I1H), 6.81 1 6.75 j=8.6 Hz, 1 4.34 2H), 4. 10 j= 13.2 Hz, 2H), 3.02 10.0 Hz, 2H), 2.82 (mn, 1H), 2.18 3H), 1.98 J=10.3 Hz, 2H), 1.83 (mn, 2H).
WO 98/00134 WO 9800134PCTIUS97/11133 228 SCHEME 34
OH
1HO 3 S /N 2 Cl 2. Na 2
S
2
O
4 34-1
OH
N)
NH
2 Boc 2 O, CHCI 3 ref lux
OH
N'
NHBoc Br COOEt 1/2 eq CS 2
CO,
3
DMF
34-2 O COQEt NHBoc HOI (gas) EtOAc, 0 0
C
*HCI
CI
NH
2 34-4 WO 98/00134 WO 9800134PCT/US97/11133 -229- SCHEME 34 (CONT'D) Boc-NN COCH Cl PF 6 Boo- No i-Pr 2 NEt, CH 2
CI
2 B Boo-N N- LiOH COOEt THF, H 2 0 34-6 cI HOI (gas) EtOAc, 0 0
C
34-7z
*HCI
/'-CQOH
WO 98/00134 PCT/US97/11133 -230- 3-Chloro-4-aminophenol (34-2) Using a procedure similar to that described for compound 12-2 but starting with 3-chlorophenol 3-chloro-4-aminophenol (34-2) was prepared. 1 H NMR (CDC13): 6 6.57 (dd, J= 2.8, 8.6 Hz, 1H), 6.73 2H).
4-(1.1 -Dimethvlethoxycarbonvl)amino-3-chlorophenol (34-3) To a 100 mL round bottomed flask with a stirring bar and an argon inlet was added 4-(1,1-dimethylethoxycarbonyl)amino-3chlorophenol (1.91 g, 13.3 mmol), di-tert-butyldicarbonate (3.14 g, 14.4 mmol) and THF (50 mL). To this mixture was added triethylamine (2.23 mL, 16.0 mmol) and the mixture was stirred at room temperature for 98 h. The solvent was removed in vacuo. The residue was acidified with 10% aqueous citric acid and extracted with EtOAc. The EtOAc extract was washed with water and brine. Drying (MgSO4), filtration, removal of the solvent in vacuo and chromatography on silica gel using EtOAc-hexane as eluant gave an oil.
This material was dissolved in THF (25 mL) and MeOH (8 mL) and IN LiOH (25 mL) were added. This solution was stirred at ambient temperature 4 h. This mixture was acidified with 10% citric acid and extracted with EtOAc. The organic layer was dried (MgSO4), filtered and concentrated in vacuo to give 34-3 as an oil. 1 H NMR (CDC13): 1.53 9H), 6.69 (dd, J= 2.8, 8.9 Hz, 2H), 6.85 J= 2.8 Hz, 1H), 7.80 J= 8.9 Hz, 1H).
Ethyl 4-(1,1-dimethylethoxycarbonyl)amino-3-chlorophenoxyacetate (34-4) To a 50 mL round bottomed flask with a stirring bar and an argon inlet was added 4-(1,1-dimethylethoxycarbonyl)amino-3chlorophenol (0.831 g, 3.41 mmol) and DMF (16 mL). This mixture was cooled in an ice bath then Cs2CO3 (0.552 g, 1.69 mmol) and ethyl bromoacetate (0.37 mL, 3.34 mmol) were added. The resulting mixture was stirred for 6 h, during which time the cooling bath was allowed to expire. The mixture was filtered through a frit and the DMF was WO 98/00134 WO 9800134PCTIUS97/11133 -231removed in vacuo. The residue was dissolved in EtOAc and washed with water (12X) and brine. Drying (MgSO4), filtration and removal of the solvent in vacio gave an oil which was purified by silica gel chromatography (EtOAc-hexane) to give ethyl 1,1-dimethylethoxycarbonyl)amino-3-chlorophenoxyacetate as an oil. I1 NMR (CDCl3): 6 1.30 (in, 3H), 1.53 9H), 4.27 (in, 2H), 4.57 (s, 2H), 6.7 8 (br s, I1H), 6.8 3 (dd, J= 2.8, 8.8 Hz, I1H), 6.96 J=2.8 Hz, 1H), 8.02 (br d, J= 8.8 Hz, I1H).
4- [4-(l1 -Piperazinyl)phenylcarbony lamino] -3 -chiorophenoxyacetic acid (34-8) Using a sequence similar to that described for compound 12-8, but starting with ethyl 4-(1,1-dimethylethoxycarbonyl)amino-3chlorophenoxyacetate, (34 (1-piperazmnyl)phenylcarbonylamnino] -3-chlorophenoxyacetic acid was prepared. IH NMR (DMSO-d6, HCl salt): 5 3.22 J= 5.0 Hz, 4H), 3.53 J= 5.0 Hz, 4H), 4.75 (s, 2H), 6.94 (dd, J= 2.9, 8.8 Hz, 1H), 7.08 8.9 Hz, 2H), 7.11 (d, J=2.9 Hz, I1H), 7.40 J=8.8 Hz, I1H), 7.91 J= 8.9 Hz, 2H), 9.12 (br s, 1H), 9.69 1H).
WO 98/00134 WO 9800134PCTIUS97/11133 -232- SCHEME Boo-N \jN-a CO Boc-N
B
2
H
6
-THF
TH F/0 0
C
MnO 2 /CHC1 3
/RT
Boo-N N-G N- CHO 35-2
OH
OH
BrCH 2 COOEt
CS
2
CO
3
DMF
0COQEt
OH
35-4 WO 98/00134 WO 9800134PCTIUS97/11133 233 SCHEME 35 (CONT'D) 35-4 Ph 3 P/CBr 4 C H 2
C[
2 /0 0
C
Ph 3 P/PhH ~COOEt PlPh 3 Br- 35-6 NaH/DMSOITH F/ U
(N)
N
Boo WO 98/00134 PCT/US97/11133 -234- SCHEME 35 (CONT'D) 1. 10% Pd-C /H 2 2. 1N NaOH/THF 3. HCI (g)/EtOAc H-N N S/ \-COOH 35-9 1 -Hydroxymethyl-4-(4-( 1,1 -dimethylethoxycarbonyl)piperazinyl)benzene (35-1) To an oven dried 500 mL round bottomed flask with a stirring bar and an argon inlet was added acid 1-5 (5.00 g, 16.32 mmol) and distilled, dry, THF (80 mL). This solution was cooled in an ice bath and borane-THF complex (125 mL of a 1M solution in THF, 125 mmol) was added with a syringe. The resulting solution was maintained at 0 0
C
for 3.5 h. The reaction was quenched by addition of MeOH (50 mL), slowly, with a syringe over 5 min. This mixture was stirred at ambient temperature for 18 h. The solvents were removed in vacuo and the residue was dissolved in EtOAc (300 mL). This solution was washed with saturated aqueous NaHCO3 and brine. Drying (MgSO4), filtration and removal of the solvent in vacuo gave 4.86 g of l-hydroxymethyl-4- (4-(1,1-dimethylethoxycarbonyl)piperazinyl)benzene, 35-1, as a white WO 98/00134 PCT/US97/11133 -235solid. 1H NMR (CDC13): 5 1.48 9H), 3.11 4H), 3.59 4H), 4.61 2H), 6.91 J=7 Hz, 2H), 7.27 J=7 Hz, 2H).
-Dimethvlethoxvcarbonyl)piperazinvl)benzaldehyde (35-2) To a 50 mL round bottomed flask with a stirring bar and an argon inlet was added 1-hydroxymethyl-4-(4-(l,1-dimethylethoxycarbonyl)piperazinyl)benzene, 35-1 (2.15 g, 7.35 mmol), CHC13 (100 mL), and MnO2 (12.0 g, 138 mmol). This mixture was stirred at ambient temperature for 24 h. The salts were removed by filtration and the solvent was removed in vacuo. The residue was chromatographed on 90 g of silica gel using 30/70 EtOAc-hexane as eluant. There was obtained 4-(4-(1,1-dimethylethoxycarbonyl)piperazinyl)benzaldehyde 35-2 (2.10 g, 99%) as a crystalline solid. 1 H NMR (CDC13): 5 1.49 (s, 9H), 3.39 4H), 3.59 4H), 6.89 J=9 Hz, 2H), 7.75 J=9 Hz, 2H), 9.79 1H).
Ethyl 2-(4-hvdroxvmethylphenoxv)acetate (35-4) To a 500 mL round bottomed flask with a stirring bar and an argon inlet was added 4-hydroxybenzyl alcohol (7.50 g, 60.42 mmol), Cs2CO3 (29.53 g, 90.63 mmol), DMF (100 mL), and ethyl bromoacetate (11.09 g, 66.46 mmol). This mixture was stirred at ambient temperature for 2 h. The mixture was filtered and the filtrate was concentrated in vacuo. The residue was partitioned between EtOAc and water. The layers were separated and the organic phase was washed with two additional portions of water. Drying (MgSO4), filtration and removal of the solvent in vacuo, gave an oil. This material was chromatographed on 250 g of silica gel using 1:1 EtOAc-hexane as eluant. There was obtained ethyl 2-(4-hydroxymethyl-phenoxy)acetate 35-4 (7.47 g, 59%) as an oil. 1 H NMR (CDC13): 5 1.29 J=7 Hz, 3H), 2.07 (br s, IH), 4.26 J=7 Hz, 2H), 4.58 2H), 4.60 2H), 6.88 J=9 Hz, 2H), 7.27 J=9 Hz, 2H).
WO 98/00134 PCT/US97/11133 -236- Ethyl 2-(4-bromomethylphenoxy)acetate (35-5) To a 1L round bottomed flask with a stirring bar, addition funnel and an argon inlet was added ethyl 2-(4-hydroxymethylphenoxy)acetate 35-4 (7.47 g, 35.53 mmol), CBr4 (13.26 g, 39.97 mmol) and dry CH2C12 (300 mL). This solution was cooled in an ice bath and and a solution of triphenyl phosphine (10.48 g, 39.97 mmol) in CH2C12 (100 mL) was added dropwise over I h. The ice bath was allowed to expire and and the mixture was stirred at ambient temperature 18 h. The solvent was removed in vacuo and the residue was chromatographed directly on 200 g of silica gel useing 15% EtOAchexane as eluant. There was obtained ethyl 2-(4-bromomethylphenoxy)acetate 35-5 (7.2 g, 74%) as a low melting solid, mp.: 44- 1 H NMR (CDC13): 8 1.29 J=7 Hz, 3H), 4.26 J=7 Hz, 2H), 4.48 2H), 4.61 2H), 6.86 J=9 Hz, 2H), 7.32 J=9 HZ, 2H).
4-(Ethoxycarbonylmethoxy)benzyltriphenylphosphonium bromide (35-6) To a 100 mL round bottomed flask with a stirring bar and a reflux condenser topped with an argon inlet was added ethyl 2-(4bromomethylphenoxy)acetate 35-5 (2.73 g, 10 mmol), triphenylphosphine (2.62 g, 10 mmol) and dry benzene (50 mL). This solution was warmed to 50°C for 20 h. The mixture was cooled to room temperature and the product was collected by filtration. The white solid was washed with a little benzene and dried in vacuo to give 4-(ethoxycarbonylmethoxy)benzyltriphenyl-phosphonium bromide 35-6 as a white, crystalline solid (4.79 g, 1 H NMR (CDC13): 5 1.28 (t, J=7 Hz, 3H), 4.27 J=7 Hz, 2H), 4.53 2H), 5.38 J=14 Hz, 2H), 6.66 J=9 Hz, 2H), 7.06 J=9 Hz, 2H), 7. 62 6H), 7.76 9H).
Ethyl 4-(2-(4-(4-(1,1-dimethylethoxycarbonyl)piperazinyl)phenyl)ethenyl)phenoxvacetate (35-7) To an oven dried 100 mL round bottomed flask with a stirring bar and an argon inlet was added 4-(ethoxycarbonylmethoxy)benzyltriphenyl-phosphonium bromide (1.106 g, 2.07 mmol) WO 98/00134 PCT/US97/11133 -237and dry THF (50 mL). To this well stirred mixture was added a solution of lithium hexamethyldisilylazide (2.20 mL of a 1M solution in THF). The deep red.solution was stirred 1 h at ambient temperature then cooled in an ice bath to 0°C. To this mixture was added a solution of -dimethylethoxycarbonyl)piperazinyl)benzaldehyde (0.60 g, 2.07 mmol) in 10 mL of THF. The ice bath was removed and the solution was stirred at ambient temperature for 3 h. The reaction mixture was diluted with EtOAc and washed with water and brine.
Drying (MgSO4), filtration and removal of the solvent in vacuo gave 1 g of an oil. This material was chromatographed on 80 g of silica gel using 25% EtOAc-hexane as eluant. There was obtained (E)-ethyl 4-(2- -dimethylethoxycarbonyl)piperazinyl)phenyl)ethenyl)phenoxyacetate 35-7 as an oil. IH NMR (CDC13): 5 1.26 (t, j=7 Hz, 3H), 1.49 9H), 3.16 4H), 3.57 4H), 4.27 j=7 Hz, 2H), 4.60 2H), 6.90 6H), 7.40 4H), and 175 mg of a mixture of E,Z isomers.
4-(2-(4-(1-Piperazinyl)phenyl)ethvl)phenoxyacetic acid (35-8) To a 25 mL round bottomed flask with a stirring bar and a balloon hydrogenation adaptor was added ethyl dimethylethoxycarbonyl)piperazinyl)phenyl)ethenyl)phenoxyacetate (175 mg, 0.36 mmol), absolute EtOH (10 mL) and 10% Pd-C. This mixture was hydrogenated at 1 atmosphere at ambient temperature for 18 h. The cataylst was removed by filtration and the filtrate was concentrated in vacuo. The residue was chromatographed on 40 g of silica gel using 20% EtOAc-hexane as eluant. The purified hydrogenation product was hydrolyzed in THF (2 mL) with 1N NaOH (1 mL) for 3 h at ambient temperature. The reaction mixture was diluted with EtOAc and washed with 10% aqueous citric acid, water, and brine. Drying (MgSO4), filtration and removel of the solvent in vacuo gave the corresponding acid as a white solid. This material was dissolved in EtOAc (20 mL) and cooled to 0°C in an ice bath. The solution was saturated with dry HC1 gas for 15 min. The resulting suspension was aged 30 min at 0°C, then the solvent and excess HC1 WO 98/00134 PCT/US97/11133 -238 were removed in vacuo. The solid product was triturated with fresh EtOAC and collected on a frit. After drying at 600C/0.05 torr there was obtained 103 mg of 4-(2-(4-(l-piperazinyl)phenyl)ethyl)phenoxyacetic acid 35-8 as the hydrochloride salt. mp 250 0 C. 1H NMR (DMSO-D6): i 2.76 4H), 3.20 4H), 3.33 4H), 4.61 2H), 6.80 J=8 Hz, 2H), 6.90 J=8 Hz, 2H), 7.12 4H), 9.21 (br s, 2H).
4-(2-(4-(1-Piperazinyl)phenyl)ethenyl)phenoxyacetic acid (35-9) To a 25 mL round bottomed flask with a stirring bar and an argon inlet was added ethyl 4-(2-(4-(4-(1,1-dimethylethoxycarbonyl)piperazinyl)phenyl)ethenyl)phenoxyacetate (57 mg, 0.12 mmol), THF (2.0 mL) and IN NaOH (1.00 mL). This mixture was stirred at ambient temperature 18 h. The reaction was neutralized with 1N HCI (1.00 mL) and extracted with several portions of EtOAc. The combined EtOAc extracts were washed with water and brine. Drying (MgSO4), filtration and removal of the solvent in vacuo gave 45 mg of a solid. This solid was dissolved in 4N HCI-1,4-dioxane. The mixture was stirred 18 h at ambient temperature. The milky suspension was concentrated in vacuo. The crude product was triturated with a little EtOAc and collected on a frit. This material was dried in vacuo to provide 24 mg of 4-(2-(4-(1-piperazinyl)phenyl)ethenyl)phenoxyacetic acid 35-9 as the hydrochloride salt, mp 250 0 C. 1 H NMR (DMSO-D6): 8 3.2 4H), 3.45 4H), 4.69 2H), 6.89 J=9 Hz, 2H), 6.98 J=9 Hz, 2H), 7.02 2H), 7.48 4H), 9.15 (br s, 2H), 13.02 (br s, 1H).
WO 98/00134 WO 9800134PCTIUS97/11 133 239 SCHEME 36 H-r4/S
NO
2 0 36-1 NaH 2
PO
4
/H
2 0
H
2 0 2 NaCIO 2
/H
2 0
CH
3
CN
HO -f S 0 0 36-2 HOI (gas) MeOH MeO rIasN0 0 36-3 PtS on C MeNH MeOH Se H n-BuOH, reflux OMe BOCN_ S NaOH, H 2 0 MeOH/ THF 36-5 2. BOC 2 0, Et 3 N, CH 2
CI
2 WO 98100134 WO 9809134PCT/US97/11133 240 SCHEME 36 (CONT'D) 0 1 C)L yCI 0 cat. DMF, CH 2
CI
2 0 DMAP, pyridine, np~t 8000
HOI
10-4 0 H -4
H
3 C 0
N
1. NaOH, H 2 0, MeOH Et 2. TFA, 0H 2 C1 2 0 ,S IN0 0
N
H
H
3 C OH WO 98/00134 PCT/US97/11133 -241- HO s NO 2 0 36-2 5-Nitrothiophene-2-carboxylic acid (36-2) To a 500 mL round bottomed flask with a stirring bar, was added 5-nitrothiophene-2-carboxaldehyde (7.86 g, 50.00 mmol) in CH- 3CN (50 mL), NaH2PO4 (1.86 g, 13.50 mmol) in water (20 mL), and H202 (aq) (6 mL). To this mixture, cooled in an ice bath, was added NaC102 (8 g, 70.80 mmol) in water (70 mL) dropwise over a period of an hour. The reaction was stirred at room temperature for hours. It was worked up by addition of Na2SO3 (500 mg), acidified with 1M HC1, and then extracted with ethyl acetate The combined organic layers were washed with water and brine Drying (MgSO4), filtration and removal of the solvents in vacuo yielded Nitrothiophene-2-carboxylic acid as a yellow powder.
1 H NMR (300 MHz, CDC13) 5 7.91(d, J= 4.4 Hz, 1H), 7.80(d, J= 4.4 Hz, 1H) MeO
NO
2 0 36-3 Methyl 5-nitrothiophene-2-carboxylate (36-3) To a 500 mL round bottomed flask with a stirring bar, was added 5-nitrothiophene-2-carboxylic acid (4.62 g, 26.68 mmol) and MeOH (100 mL). This solution was cooled in an ice bath and dry HCI gas was bubbled through the solution till saturated. The reaction was equipped with a condensor and a drying tube and refluxed overnight.
The solvent was then removed in vacuo to a yellow solid, which was WO 98/00134 PCT/US97/11133 -242then dissolved in EtOAc. This solution was washed with NaHCO3 (sat, aq) and brine dried (MgSO4), and filtered. Removal of the solvents in vacuo and drying of the product in vacuo yielded methyl nitrothiophene-2-carboxylate as a tan solid.
1H NMR (300 MHz, CDC13) 8 7.88(d, J= 4.4 Hz, 1H), 7.70(d, J= 4.4 Hz, 1H), 3.96(s, 3H) Meo
NH
2 0 36-4 Methyl 5-aminothiophene-2-carboxvlate (36-4) To a Parr flask was added methyl 5-nitrothiophene-2carboxylate (5.00 g, 26.71 mmol) and MeOH (100 mL). The solution was purged with a stream of Ar, and 5% platinum on sulfide carbon (2.00 g) was added. The mixture was hydrogenated on a Parr apparatus set at 50 psi for 6.5 hours. The reaction was then filtered through celite and the solvents removed in vacuo to yield methyl 5-aminothiophene-2carboxylate as an olive green oil.
1 H NMR (300 MHz, CDC13) 8 7.45(d, J= 4.0 Hz, 1H), 6.09(d, J= Hz, IH), 4.31(br s, 2H), 3.81(s, 3H) r^ SN OMe BOCNS
O-
-Dimethylethoxycarbonyl)-piperazin- -yl)-thiophene-2carboxylate. (36-5) To a 500 mL round bottomed flask equipped with a condenser, stirring bar, and argon inlet was added methyl aminothiophene-2-carboxylate (4.27 g, 27.16 mmol), bis(2- WO 98/00134 PCT/US97/11133 -243chloroethyl)amine, hydrochloride (5.82 g, 32.59 mmol) and n-BuOH mL, previously purged with Ar). This solution was reluxed for 7 days under Ar, and the reaction was followed by HPLC. The solvents were then removed in vacuo, and the residue partitioned between EtOAc and NaHCO3 Extracted aqueous with EtOAc and then washed the combined organics with brine dried (Na2SO4), filtered, and concentrated in vacuo. The residue was azeotroped with toluene and dried in vacuo overnight.
The dried product was dissolved in CH2C12 (80 mL), and triethylamine (4.2 mL, 30.13 mmol), di-tert-butylpyrocarbonate (6.53 g, 29.90 mmol), and CH2C12 (20 mL) were added and the reaction was stirred at room temperature for 2 hours. The reaction was then concentrated in vacuo. This yielded 490 mg of n-butyl dimethylethoxycarbonyl)-piperazin-1 -yl)-thiophene-2-carboxylate and 700 mg of ,-dimethylethoxycarbonyl)-piperazin-l-yl)thiophene-2-carboxylate as a mixture of the methyl and n-butyl esters.
1 H NMR of n-butyl ester (300 MHz, CDC13) 8 7.55(d, J= 4.4 Hz, 1H), 6.06(d, J= 4.4 Hz, 1H), 4.24(t, 2H), 3.58(t, 4H), 3.22(t, 4H), 1.69(m, 2H), 1.48(s, 9H), 1.42(m, 2H), 0.95(t, 3H) BOcN s
OH
BOCN\J 0 36-6 -Dimethylethoxycarbonyl)-piperazin- -yl)-thiophene-2carboxylic acid, (36-6) To a 50 ml round bottomed flask with a stirring bar was added n-butyl 5-(4-(1,1-dimethylethoxycarbonyl)-piperazin-1-yl)thiophene-2-carboxylate (490 mg, 1.33 mmol) and 4.2 mL of a NaOH solution in 3:1 MeOH/H20. THF (2 mL) was added to keep the reaction homogeneous. The reaction was stirred atroom temperature and followed by HPLC. After 2 days, the reaction was concentrated in WO 98/00134 PCT[US97/11133 -244vacuo acidified with 10% citric acid and extracted with EtOAc (3X).
The combined organics were washed with water brine dried (Na2SO4), filtered, and the solvents removed in vacuo. The product was azeotroped with benzene (3X) and dried in vacuo over 2 days to yield 5-(4-(1,1-dimethylethoxycarbonyl)-piperazin-1-yl)-thiophene-2carboxylic acid as a dark green solid.
1 H NMR (300 MHz, CD30D) 8 7.51(d, J= 4.4 Hz, 1H), 6.19(d, J=4.4 Hz, 1H), 3.57(t, 4H), 3.24(t, 4H), 1.47(s, 9H) NS N-P-0 0 BOCN/ HaC OEt 2 Ethyl 2-(4-(5-(4-(1,1-dimethylethoxycarbonyl)-piperazin-1 -yl)-2thienylcarbonylamino)-3-methylphenoxv)-acetate (36-7) To a 100 ml round bottomed flask with a stirring bar and an argon inlet was added 5-(4-(1,1-dimethylethoxycarbonyl)-piperazin- 1-yl)-thiophene-2-carboxylic acid (428 mg, 1.370 mmol), anhydrous CH2C12 (10 mL), DMF (.05 mL), and distilled oxalyl chloride (.180 mL, 2.06 mmol). It was stirred 1.5 hours at room temperature. The acid chloride was concentrated in vacuo azeotroped with benzene (3X), and stored under a blanket of Ar.
To another 100 mL round bottomed flask with a stirring bar and an argon inlet was added ethyl 4-amino-3-methylphenoxyacetate hydrochloride, anhydrous pyridine (7 mL), and dimethylaminopyridine (34 mg, 0.278 mmol).
Cooled in an ice bath. A solution of the acid chloride in pyridine (3 mL) was added. The ice bath was removed and the reaction was stirred at 80°C overnight.
WO 98/00134 WO 9800134PCT/US97/1 1133 -245- The reaction was concentrated in vacuo dissolved in EtOAc, washed with aq. KHS04 aq. NaHCO3 brine (LX), dried (Na2SO4), filtered, and the solvents removed in vacuo. This material was chromatographed on silica gel in 18% EtOAc-hexane as eluant yielding ethyl 1,1 -dimethylethoxycarbonyl)piperazin- 1 -yl)-2-thienylcarbonylamino)-3 -methylphenoxy)-acetate.
IH NMR (300 MHz, DMSO-d6) 5 9.42(s, IH), 7.67(d, J= 4.3 Hz, LH), 7.13(d, 1= 8.8 Hz, IH), 6.83(d, J=2.7 Hz, LH), 6.74(dd, J= 8.8, 2.7 Hz, IH), 6.24(d, J= 4.3 Hz, 1H), 4.76(s, 2H), 4.17(q, J= 7.1, 2H), 3.47(t, 4H), 3.16(t, 4H), 2.15(s, 3H), 1.42(s, 9H), 1.22(t, J= 7.1 Hz, 3H)
NJH
3 0 OH
H
36-8 1,1 -Dimethylethoxycarbonyl)-piperazin- l-yl)-2thienylcarbonylamino)-3 -methvlphenoxy)-acetic acid (36-8) To a 100 mnL round bottomed flask with a stirring bar was added ethyl 1,1 -dimethylethoxycarbonyl)-piperazin- l-yl)-2thienylcarbonylamino)-3 -methylphenoxy)-acetate (250mg, 0.496 mimol), MeOH (8 mL), and IM NaOH (1 mL). The reaction was stirred overnight at room temperature.
The reaction was concentrated in vacuc, acidified with aq.
KHS04 and extracted with EtOAc The organic extracts were washed with brine dried (Na2SO4), filtered, and the solvents removed in vacuc.
This product was dissolved in CH2C12 (8 mL), and trifluoroacetic acid (2 mL) was added. The reaction was stirred for 2 hours at room temperature and then concentrated in vacuo. The residue was subjected to HPLC on a G- 18 reverse-phase column eluting with WO 98/00134 PCT/US97/11133 -246- 100% H20 -50% H20/CH3CN (with 0.1% TFA) over 40 minutes.
Collection and lyophilization of the appropriate fractions yielded 2-(4- -Dimethylethoxycarbonyl)-piperazin-1 -yl)-2thienylcarbonylamino)-3-methylphenoxy)-acetic acid as a white fluffy solid.
1 H NMR (300 MHz, CD30D) 8 7.62(d, J= 4.1 Hz, 1H), 7.16(d, J= Hz, 1H), 6.86(d, J=2.7 Hz, 1H), 6.79(dd, J= 8.5, 2.7 Hz, 1H), 6.34(d, J= 4.1 Hz, 1H), 4.65(s, 2H), 3.50(m, 4H), 3.40(m, 4H), 2.24(s, 3H) EXAMPLE 37 Tablet Preparation Tablets containing 25.0, 50.0, and 100.0 mg., respectively, of the active compound 3-(4-(4-piperazin-1-ylphenylcarbonylamino)phenyl)-propanoic acid are prepared as illustrated below: TABLE FOR DOSES CONTAINING FROM 25-100MG OF THE ACTIVE COMPOUND Amount-mg Active Compound 25.0 50.0 100.0 Microcrystalline cellulose 37.25 100.0 200.0 Modified food corn starch 37.25 4.25 Magnesium stearate 0.50 0.75 All of the active compound, cellulose, and a portion of the corn starch are mixed and granulated to 10% corn starch paste. The resulting granulation is sieved, dried and blended with the remainder of the corn starch and the magnesium stearate. The resulting granulation is then compressed into tablets containing 25.0, 50.0, and 100.0 mg, respectively, of active ingredient per tablet.
WO 98/00134 PCT/US97/11133 -247- EXAMPLE 38 Intravenous formulations An intravenous dosage form of the above-indicated active compound is prepared as follows: Active Compound 0.5-10.0mg Sodium Citrate 5-50mg Citric Acid 1-15mg Sodium Chloride 1-8mg Water for Injection (USP) q.s. to 1 L Utilizing the above quantities, the active compound is dissolved at room temperature in a previously prepared solution of sodium chloride, citric acid, and sodium citrate in Water for Injection (USP, see page 1636 of United States Pharmacopeia/National Formulary for 1995, published by United States Pharmacopeial Convention, Inc., Rockville, Maryland, copyright 1994.
EXAMPLE 39 Intravenous formulation A pharmaceutical composition was prepared at room temperature using 3-(4-(4-piperazin-1-ylphenylcarbonylamino)phenyl) propanoic acid, a citrate buffer, and sodium chloride, to obtain a concentration of of 0.25 mg/ml.
800 grams of water was introduced into a standard pharmaceutical mixing vessel. 0.25 grams of 4-(N-piperazine)benzoyl- N-(aminophen-4-yl)propionic acid was dissolved in the water. 2.7 grams sodium citrate and 0.16 grams citric acid were added to obtain a finished citrate concentration of 10 mM. 8 grams of sodium chloride was added. 200 grams of water was then added to achieve the desired WO 98/00134 PCT/US97/11133 -248final concentrations of ingredients. The resulting aqueous formulation had the following concentrations: Ingredient Amount 3-(4-(4-piperazin-1-ylphenylcarbonylamino)phenyl) propanoic acid 0.25 mg/ml citrate buffer 10 mM sodium chloride 8 mg/ml The finished concentrated formulation is stored in a standard USP Type I borosilicate glass container at 30-40 degrees C.
Prior to compound administration, the concentrated formulation is diluted in a 4:1 ratio resulting in a finished concentration of 0.05 mg/ml and transfered to an infusion bag, Therapeutic Treatment Compounds of the invention may be administered to patients where inhibition of human or mammalian platelet aggregation or adhesion is desired.
Compounds of the invention are useful in inhibiting platelet aggregation and thus, they may find utility in surgery on peripheral arteries (arterial grafts, carotid endaterectomy) and in cardiovascular surgery where manipulation of arteries and organs, and/or the interation of platelets with artificial surfaces, leads to platelet aggregation and consumption. The aggregated platelets may form thrombi and thromboemboli. Compounds of the invention may be administered to these surgical patients to prevent the formation of thrombi and thromboemboli.

Claims (31)

1. A compound having the formula X-Y-Z-A-B and pharmaceutically acceptable salts, X is H N HN Nj N H 2 N N CN-j N 1 H 2 N H-D- CH 3 Y is N or X and Y form the fused S 0@ 0 BS Sd 0@ .5 S. OOS S *5 0e S S. S *5S S S. *0 6O 0 6 0 *050 5 S 0 6 S S. S SO S SS 0 .6 5* S HN [R:\LIBAA]07996.doc~tab Z is 250 0 0 11 11 -CNH-'7 -CH 2 CH 2 0 0 11 11 o -CH=CH-' -NHC or GH 3 Z represents a bond; A is NHSO 2 C 6 H 5 OH 3 OCH 3 CF 3 a a a a a *aa. a OH 3 OH 3 OH 3 N0 2 OCH 3 Br OH 3 Br and B is -OCH 2 C0 2 H, [R:\LIBAA]07996.doc tab -OCH 2 00 2 CH 2 C(O)NHCH 3 -OCH 2 CO 2 CH 2 C(O)N(0H 2 0H3)2, -OCH 2 00 2 CH 2 CH 3 -CH 2 CH 2 CO 2 H, -CH 2 COOH, -OCH(0H 3 )00 2 H, or -OCH(CH 3 )CO 2 CH 2 CH 3
2. A compound selected from the group consisting of
3-(4-(4-piperazin-1 -ylphenylcarbonylamino)phenyl)propanoic acid, 2-(4-(4-Piperazinyl-1 -yl)phenylcarbonylamino)phenoxy)acetic acid, Ethyl -piperazinyl)phenylcarbonylamino)pheloxy)acetate, hydrochloride, -Piperazinyl)phenylcarbonylamino)indoI-I-yl)acetic acid, 3-(3-(4-Piperazin-1 -ylphenyl)carbonylamino)phelyl)propanoic acid, Ethyl 2-(4-(4-(piperazin-1 -yl)phenylcarbonylamino)phenoxy)propafloate, 2-(4-(4-(Piperazin-1 -yl)phenylcarbonylamino)phenoxy)propionic acid, 2-(4-(((2-Piperazin-1 -yI)pyridin-5-yl)carbonylamino)pheloxy)acetic acid, 3-Methyl-4-((1 ,2,3,4-tetrahyd ro-9H-pyrido[3,4-b]indol-7- yI)carbonylamino)phenoxyacetic acid, ,1 -dimethylethoxycarbonyl)-piperazin-1 -yl)-2-thienylcarbonylamino)-3- methylphenoxy)-acetic acid, 00 HN N 0 -C0
4-((2,3,4,5-tetrahydropyrazino-[1 ,2-a]indole-8-yI)carbonylamino)-3- methylphenoxyacetic acid, and di methyl ethoxyca rbonyl)- 1 1 a,2,3,4,5-hexahyd ropyrazilo-[1, 2- a]indole-8-yl)carbonylamino)-3-methylpheloxyacetic acid, and pharmaceutically acceptable salts thereof. A fibrinogen receptor antagonist, substantially as hereinbefore described with reference to any one of the examples. 4. A composition comprising a compound of any one of claims 1 to 3 and a pharmaceutically acceptable carrier. [R:\LIBAA)07996.doctab 252 A composition of claim 5, further comprising an efficacious amount of one or more agents selected from a thrombolytic agent, an anticoagulant agent, and an antiplatelet agent and a pharmaceutically acceptable carrier.
6. A method for the treatment or prophylaxis of thrombus formation in a mammal, which method includes or consists of administering to said mammal an effective amount of at least one compound according to any one of claims 1 to 3, or of a composition according to claim 4.
7. A compound according to any one of claims 1 to 3 or a composition according to claim 4 when used in the treatment or prophylaxis of thrombus formation.
8. Use of a compound according to any one of claims 1 to 3 for the manufacture of a medicament for the treatment or prophylaxis of thrombus formation.
9. A method for the treatment or prophylaxis of embolus formation in a mammal, which method includes or consists of administering to said mammal an effective amount of at least one compound according to any one of claims 1 to 3, or of a composition according to claim 4. A compound according to any one of claims 1 to 3 or a composition S according to claim 4 when used in the treatment or prophylaxis of embolus S: 20 formation.
11. Use of a compound according to any one of claims 1 to 3 for the manufacture of a medicament for the treatment or prophylaxis of embolus i: formation.
12. A method for inhibiting the binding of fibrinogen to blood platelets in a 25 mammal, which method includes or consists of administering to said mammal an effective amount of at least one compound according to any one of claims 1 to 3, or of a composition according to claim 4.
13. A compound according to any one of claims 1 to 3 or a composition according to claim 4 when used in inhibiting the binding of fibrinogen to blood 30 platelets.
14. Use of a compound according to any one of claims 1 to 3 for the manufacture of a medicament for inhibiting the binding of fibrinogen to blood platelets. A method for inhibiting the aggregation of blood platelets in a mammal, <757 which method includes or consists of administering to said mammal an effective [R:\LIBAA]07996.doc:tab 253 amount of at least one compound according to any one of claims 1 to 3, or a composition according to claim 4.
16. A compound according to any one of claims 1 to 3 or a composition according to claim 4 when used in inhibiting the aggregation of blood platelets.
17. Use of a compound according to any one of claims 1 to 3 for the manufacture of a medicament for inhibiting the aggregation of blood platelets.
18. A method for inhibiting osteoclast mediated bone resorption in a mammal, which method includes or consists of administering to said mammal an effective amount of at least one compound according to any one of claims 1 to 3, or to of a composition according to claim 4.
19. A compound according to any one of claims 1 to 3 or a composition according to claim 4 when used in inhibiting osteoclast mediated bone resorption. Use of a compound according to any one of claims 1 to 3 for the manufacture of a medicament for inhibiting osteoclast mediated bone resorption.
21. A method for inhibiting angiogenesis in a mammal, which method includes or consists of administering to said mammal an effective amount of at least one compound according to any one of claims 1 to 3, or of a composition according to claim 4.
22. A compound according to any one of claims 1 to 3 or a composition 20 according to claim 4 when used in inhibiting angiogenesis. g: 23. Use of a compound according to any one of claims 1 to 3 for the manufacture of a medicament for inhibiting angiogenesis.
24. A method for inhibiting tumour growth in a mammal, which method includes or consists of administering to said mammal an effective amount of at least 25 one compound according to any one of claims 1 to 3, or of a composition according to claim 4.
25. A compound according to any one of claims 1 to 3 or a composition according to claim 4 when used in inhibiting tumour growth.
26. Use of a compound according to any one of claims 1 to 3 for the :i 30 manufacture of a medicament for inhibiting tumour growth.
27. A method for preventing platelet thrombosis in a mammal, which method includes or consists of administering to said mammal an effective amount of at least one compound according to any one of claims 1 to 3, or of a composition according to claim 4. T 28. A compound according to any one of claims 1 to 3 or a composition according to claim 4 when used in preventing platelet thrombosis. [R:\LIBAAj07996.doc:tab
29. Use of a compound according to any one of claims 1 to 3 for the manufacture of a medicament for preventing platelet thrombosis. A method for preventing thromboembolism in a mammal, which method includes or consists of administering to said mammal an effective amount of at least one compound according to any one of claims 1 to 3, or of a composition according to claim 4.
31. A compound according to any one of claims 1 to 3 or a composition according to claim 4 when used in preventing thromboembolism.
32. Use of a compound according to any one of claims 1 to 3 for the manufacture of a medicament for preventing thromboembolism.
33. A method for preventing reocclusion in a mammal, which method includes or consists of administering to said mammal an effective amount of at least one compound according to any one of claims 1 to 3, or of a composition according to claim 4.
34. A compound according to any one of claims 1 to 3 or a composition according to claim 4 when used in preventing reocclusion. Use of a compound according to any one of claims 1 to 3 for the manufacture of a medicament for preventing reocclusion.
36. A method for inhibiting the aggregation of blood platelets by blocking o fibrinogen from acting at its receptor site in a mammal, which method includes or consists of administering to said mammal an effective amount of at least one compound according to any one of claims 1 to 3 in combination with one or more agents selected from a thrombolytic agent, an anticoagulant agent, and an antiplatelet agent, or of a composition according to claim
37. A compound according to any one of claims 1 to 3 in combination with one or more agents selected from a thrombolytic agent, an anticoagulant agent, and an antiplatelet agent or a composition according to claim 5 when used in inhibiting the aggregation of blood platelets by blocking fibrinogen from acting at its receptor site. 30 38. Use of a compound according to any one of claims 1 to 3 in combination with one or more agents selected from a thrombolytic agent, an anticoagulant agent, and an antiplatelet agent for the manufacture of a medicament for inhibiting the aggregation of blood platelets by blocking fibrinogen from acting at its receptor site.
39. A method for inhibiting the binding of fibrinogen to blood platelets by ,iNblocking fibrinogen from acting at is receptor site in a mammal, which method [R:\LIBAA]07996.doc:tab 255 includes or consists of administering to said mammal an effective amount of at least one compound according to any one of claims 1 to 3 in combination with one or more agents selected from a thrombolytic agent, an anticoagulant agent, and an antiplatelet agent, or of a composition according to claim s 40. A compound according to any one of claims 1 to 3 in combination with one or more agents selected from a thrombolytic agent, an anticoagulant agent, and an antiplatelet agent or a composition according to claim 5 when used in inhibiting the binding of fibrinogen to blood platelets by blocking fibrinogen from acting at its receptor site.
41. Use of a compound according to any one of claims 1 to 3 in combination with one or more agents selected from a thrombolytic agent, an anticoagulant agent, and an antiplatelet agent for the manufacture of a medicament for inhibiting the binding of fibrinogen to blood platelets by blocking fibrinogen from acting at its receptor site. Dated 17 April, 2000 Merck Co., Inc. Patent Attorneys for the Applicant/Nominated Person SPRUSON FERGUSON 9 9 [R:\L1BAA]07996.doc:tab
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