OA11341A - Vitronectin receptor antagonists. - Google Patents
Vitronectin receptor antagonists. Download PDFInfo
- Publication number
- OA11341A OA11341A OA1200000079A OA1200000079A OA11341A OA 11341 A OA11341 A OA 11341A OA 1200000079 A OA1200000079 A OA 1200000079A OA 1200000079 A OA1200000079 A OA 1200000079A OA 11341 A OA11341 A OA 11341A
- Authority
- OA
- OAPI
- Prior art keywords
- dihydro
- dibenzo
- cycloheptene
- compound
- acetic acid
- Prior art date
Links
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D213/00—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
- 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
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P19/00—Drugs for skeletal disorders
- A61P19/08—Drugs for skeletal disorders for bone diseases, e.g. rachitism, Paget's disease
- A61P19/10—Drugs for skeletal disorders for bone diseases, e.g. rachitism, Paget's disease for osteoporosis
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P29/00—Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
- A61P35/04—Antineoplastic agents specific for metastasis
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P43/00—Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P9/00—Drugs for disorders of the cardiovascular system
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P9/00—Drugs for disorders of the cardiovascular system
- A61P9/10—Drugs for disorders of the cardiovascular system for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D213/00—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
- 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
- C07D213/74—Amino or imino radicals substituted by hydrocarbon or substituted hydrocarbon radicals
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D217/00—Heterocyclic compounds containing isoquinoline or hydrogenated isoquinoline ring systems
- C07D217/22—Heterocyclic compounds containing isoquinoline or hydrogenated isoquinoline ring systems 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 nitrogen-containing ring
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D233/00—Heterocyclic compounds containing 1,3-diazole or hydrogenated 1,3-diazole rings, not condensed with other rings
- C07D233/04—Heterocyclic compounds containing 1,3-diazole or hydrogenated 1,3-diazole rings, not condensed with other rings having one double bond between ring members or between a ring member and a non-ring member
- C07D233/28—Heterocyclic compounds containing 1,3-diazole or hydrogenated 1,3-diazole rings, not condensed with other rings having one double bond between ring members or between a ring member and a non-ring member 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
- C07D233/44—Nitrogen atoms not forming part of a nitro radical
- C07D233/48—Nitrogen atoms not forming part of a nitro radical with acyclic hydrocarbon or substituted acyclic hydrocarbon radicals, attached to said nitrogen atoms
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D239/00—Heterocyclic compounds containing 1,3-diazine or hydrogenated 1,3-diazine rings
- C07D239/02—Heterocyclic compounds containing 1,3-diazine or hydrogenated 1,3-diazine rings not condensed with other rings
- C07D239/06—Heterocyclic compounds containing 1,3-diazine or hydrogenated 1,3-diazine rings not condensed with other rings having one double bond between ring members or between a ring member and a non-ring member
- C07D239/08—Heterocyclic compounds containing 1,3-diazine or hydrogenated 1,3-diazine rings not condensed with other rings having one double bond between ring members or between a ring member and a non-ring member with hetero atoms directly attached in position 2
- C07D239/12—Nitrogen atoms not forming part of a nitro radical
- C07D239/14—Nitrogen atoms not forming part of a nitro radical with only hydrogen atoms, hydrocarbon or substituted hydrocarbon radicals, attached to said nitrogen atoms
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D263/00—Heterocyclic compounds containing 1,3-oxazole or hydrogenated 1,3-oxazole rings
- C07D263/52—Heterocyclic compounds containing 1,3-oxazole or hydrogenated 1,3-oxazole rings condensed with carbocyclic rings or ring systems
- C07D263/54—Benzoxazoles; Hydrogenated benzoxazoles
- C07D263/58—Benzoxazoles; Hydrogenated benzoxazoles 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 in position 2
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D277/00—Heterocyclic compounds containing 1,3-thiazole or hydrogenated 1,3-thiazole rings
- C07D277/02—Heterocyclic compounds containing 1,3-thiazole or hydrogenated 1,3-thiazole rings not condensed with other rings
- C07D277/20—Heterocyclic compounds containing 1,3-thiazole or hydrogenated 1,3-thiazole rings not condensed with other rings having two or three double bonds between ring members or between ring members and non-ring members
- C07D277/32—Heterocyclic compounds containing 1,3-thiazole or hydrogenated 1,3-thiazole rings not condensed with other rings having two or three double bonds between ring members or between ring members and non-ring members 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
- C07D277/38—Nitrogen atoms
- C07D277/42—Amino or imino radicals substituted by hydrocarbon or substituted hydrocarbon radicals
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D313/00—Heterocyclic compounds containing rings of more than six members having one oxygen atom as the only ring hetero atom
- C07D313/02—Seven-membered rings
- C07D313/06—Seven-membered rings condensed with carbocyclic rings or ring systems
- C07D313/10—Seven-membered rings condensed with carbocyclic rings or ring systems condensed with two six-membered rings
- C07D313/14—[b,f]-condensed
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D405/00—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom
- C07D405/02—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom containing two hetero rings
- C07D405/06—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom containing two hetero rings linked by a carbon chain containing only aliphatic carbon atoms
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D405/00—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom
- C07D405/02—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom containing two hetero rings
- C07D405/12—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom containing two hetero rings linked by a chain containing hetero atoms as chain links
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D487/00—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, not provided for by groups C07D451/00 - C07D477/00
- C07D487/02—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, not provided for by groups C07D451/00 - C07D477/00 in which the condensed system contains two hetero rings
- C07D487/04—Ortho-condensed systems
Abstract
Compounds of formula (I) are disclosed which are vitronectin receptor antagonists and are useful in the treatment of osteoporosis wherein: A is CH2 or O; R<1> is H, halo or C1-6alkyl; R<2> is H, C1-6alkyl or CH2NR''R''; X is O or CH2; Y is (a), (b), (c), (d), (e), (f) or (g); G is NR'', S or O; R' is H, C1-6alkyl, OC1-6alkyl, SC1-6alkyl, NR''R'' or halo; each R'' independently is H or C1-6alkyl; and s is 0, 1 or 2; or a pharmaceutically acceptable salt thereof.
Description
.,. 011341
TITLE
Vitronectin Receptor Antagoniste
FIELD OF THE INVENTION 5 This invention relates to pharmaceutically active compounds which inhibit the vitronectin receptor and are useful for the treatment of inflammation, cancer andcardiovascular disorders, such as atherosclerosis and restenosis, and diseases wherein bonerésorption is a factor, such as osteoporosis.
10 BACKGROUND OF THE INVENTION
Integrins are a superfamily of cell adhesion receptors, which are transmembrane glycoproteins expressed on a variety of cells. These cell surface adhesion receptors includegpllb /Hla (the fibrinogen receptor) and av^3 vitronectin receptor). The fibrinogenreceptor gpllb /Ilia is expressed on the platelet surface, and médiates platelet aggregation 15 and the formation of a hemostatic clôt at the site of a bleeding wound. Philips, et al.,
Blood., 1988, 71, 831. The vitronectin receptor av^3 ’s expressed on a number of cells,including endothélial, smooth muscle, osteoclast, and tumor cells, and, thus, it has a varietyof functions. The ανβ} receptor expressed on the membrane of osteoclast cells médiates theadhesion of osteoclasts to the bone matrix, a key step in the bone résorption process. Ross, 20 et al., J. Biol. Chem., 1987, 262, 7703. A disease characterized by excessive bone résorption is osteoporosis. The av^3 receptor expressed on human aortic smooth musclecells médiates their migration into neointima, a process which can lead to restenosis afterpercutaneous coronary angioplasty. Brown, et al., Cardiovascular Res., 1994, 28, 1815.Additionally, Brooks, et al., Cell, 1994, 79, 1157 has shown that an avB3 antagonist is able 25 to promote tumor régression by inducing apoptosis of angiogenic blood vessels. Thus,agents that block the vitronectin receptor would be useful in treating diseases, such asosteoporosis, restenosis and cancer.
The vitronectin receptor is now known to refer to three different integrins,designated avBj, avB3 and ανθ5· Horton, et al., Int. J. Exp. Pathol., 1990, 71, 741. avBj 30 binds fibronectin and vitronectin. av^3 binds a large variety of ligands, including fibrin,fibrinogen, laminin, thrombospondin, vitronectin, von Willebrand's factor, osteopontin andbone sialoprotein I. avB5 binds vitronectin. The vitronectin receptor avB5 has been shownto be involved in cell adhesion of a variety of cell types, including microvascularendothélial cells, (Davis, et al., J. Cell. Biol., 1993,51, 206), and its rôle in angiogenesis 35 has been confirmed. Brooks, et al., Science, 1994, 264, 569. This integrin is expressed onblood vessels in human wound granulation tissue, but not in normal skin. 011341 -2-
The vitronectin receptor is known to bind to bone matrix proteins which contain thetri-peptide Arg-Gly-Asp (or RGD) motif. Thus, Horton, et al., Exp. Cell Res. 1991, /95,368, disclose that RGD-containing peptides and an anti-vitronectin receptor antibody(23C6) inhibit dentine résorption and cell spreading by osteoclasts. In addition, Sato, et al., 5 J. Cell Biol. 1990, ///, 1713 discloses that echistatin, a snake venom peptide whichcontains the RGD sequence, is a potent inhibitor of bone résorption in tissue culture, andinhibits attachment of osteoclasts to bone.
It has now been discovered that certain compounds are potent inhibitors of thedCyO} and avB5 receptors. In particular, it has been discovered that such compounds are 10 more potent inhibitors of the vitronectin receptor than the fibrinogen receptor.
SUMMARY OF THE INVENTION
This invention comprises compounds of the formula (I) as described hereinafter,which hâve pharmacological activity for the inhibition of the vitronection receptor and are 15 useful in the treatment of inflammation, cancer and cardiovascular disorders, such asatherosclerosis and restenosis, and diseases wherein bone résorption is a factor, such asosteoporosis.
This invention is also a pharmaceutical composition comprising a compoundaccording to formula (I) and a pharmaceutically carrier. 20 This invention is also a method of treating diseases which are mediated by the vitronectin receptor. In a particular aspect, the compounds of this invention are useful fortreating atherosclerosis, restenosis, inflammation, cancer and diseases wherein bonerésorption is a factor, such as osteoporosis.
25 DETAILED DESCRIPTION
This invention comprises novel compounds which are more potent inhibitors of thevitronectin receptor than the fibrinogen receptor. The novel compounds comprise adibenzocycloheptene core in which a nitrogen-containing substituent is présent on one ofthe aromatic six-membered rings of the dibenzocycloheptene and an aliphatic substituent 30 containing an acidic moiety is présent on the seven-membered ring of the dibenzocycloheptene. The dibenzocycloheptene ring system is believed to orient thesubstituent sidechains on the six and seven membered rings so that they may interactfavorablÿ with the vitronectin receptor. It is preferred that about twelve to fourteenintervening covalent bonds via the shortest intramolecular path will exist between the 35 acidic group on the aliphatic substituent of the seven-membered ring of the dibenzocycloheptene and the nitrogen of the nitrogen-containing substituent on one of thearomatic six-membered ring of the dibenzocycloheptene. 011341 - 3-
This invention comprises compounds of formula (I):
10 wherein: A is CH2 or O; R1 is H, halo orCpéalkyl; R2 is H, Ci.6alkyl or CH2NR"R";X is O or CH2;
Yis
15 20 25 G is NR", S or O; R’is H, Ci.6alkyl, OCi.6alkyl, SCi_6alkyl, NR"R" or halo;each R" independently is H or Cj^alkyl; ands is 0, 1 or 2; or a pharmaceutically acceptable sait thereof.
Also included in this invention are pharmaceutically acceptable addition salts and complexes of the compounds of this invention. In cases wherein the compounds of thisinvention may hâve one or more chiral centers, unless specified, this invention includeseach unique nonracemic compound which may be synthesized and resolved byconvêntional techniques. In cases in which compounds hâve unsaturated carbon-carbon 011341 5 10 15 20 25 30 35 -4- double bonds, both the cis (Z) and trans (E) isomers are within the scope of this invention.
In cases wherein compounds may exist in tautomeric forms, such as keto-enol tautomers, O OR' such as and , and each tautomeric form is contemplated as being included within this invention whether existing in equilibrium or Iocked in one form by appropriatesubstitution with R’.
The compounds of formula (I) inhibit the binding of vitronectin and other RGD-containing peptides to the vitronectin receptor. Inhibition of the vitronectin receptor onosteoclasts inhibits osteoclastic bone résorption and is useful in the treatment of diseaseswherein bone résorption is associated with pathology, such as osteoporosis andosteoarthritis.
In another aspect, this invention is a method for stimulating bone formation whichcomprises administering a compound which causes an increase in osteocalcin release.Increased bone production is a clear benefit in disease States wherein there is a deficiencyof mineralized bone mass or remodeling of bone is desired, such as fracture healing and theprévention of bone fractures. Diseases and metabolic disorders which resuit in loss of bonestructure would also benefit from such treatment. For instance, hyperparathyroidism,Paget’s disease, hypercalcemia of malignancy, osteolytic lésions produced by bonemetastasis, bone loss due to immobilization or sex hormone deficiency, Behçet’s disease,osteomalacia, hyperostosis and osteopetrosis, could benefit from administering a compoundof this invention.
Additionally, since the compounds of the instant invention inhibit vitronectinreceptors on a number of different types of cells, said compounds would be useful in thetreatment of inflammatory disorders, such as rheumatoid arthritis and psoriasis, andcardiovascular diseases, such as atherosclerosis and restenosis. The compounds of Formula(I) of the présent invention may be useful for the treatment or prévention of other diseasesincluding, but not limited to, thromboembolie disorders, asthma, allergies, adult respiratorydistress syndrome, graft versus host disease, organ transplant rejection, septic shock,eczema, contact dermatitis, inflammatory bowel disease, and other autoimmune diseases.The compounds of the présent invention may also be useful for wound healing.
The compounds of the présent invention are also useful for the treatment, includingprévention, of angiogenic disorders. The term angiogenic disorders as used herein includesconditions involving abnormal neovascularization. Where the growth of new blood vesselsis the cause of, or contributes to, the pathology associated with a disease, inhibition ofangiogenisis will reduce the deleterious effects of the disease. An example of such adisease target is diabetic retinopathy. Where the growth of new blood vessels is required tosupport growth of a deleterious tissue, inhibition of angiogenisis will reduce the bloodsupply to the tissue and thereby contribute to réduction in tissue mass based on blood -5- 011341 supply reauirements. Examples include growth of tumors where neovascularization is acontinuai requirement in order that the tumor grow and the establishment of solid tumormétastasés. Thus, the compounds of the présent invention inhibit tumor tissueangiogenesis, thereby preventing tumor metastasis and tumor growth. 5 Thus, according to the methods of the présent invention, the inhibition of angiogenesis using the compounds of the présent invention can ameliorate the symptoms ofthe disease, and, in some cases, can cure the disease.
Another therapeutic target for the compounds of the instant invention are eyediseases chacterized by neovascularization. Such eye diseases include comeal neovascular 10 disorders, such as comeal transplantation, herpetic keratitis, luetic keratitis, pterygium andneovascular pannus associated with contact Iens use. Additional eye diseases also includeage-related macular degeneration, presumed ocular histoplasmosis, retinopathy ofprematurity and neovascular glaucoma.
This invention further provides a method of inhibiting tumor growth which 15 comprises administering stepwise or in physical combination a compound of formula (I)and an antineoplastic agent, such as topotecan and cisplatin.
With respect to formula (I):
Suitably Y is
NR“-
20 , wherein R’ is H, Cj^alkyl, OCj^alkyl,SCj^alkyl, NR"R" or Cl and each R" independently is H or Ci^alkyl.
Altemately, Y is
R"R"N
, wherein each R" is H or Cj^alkyl. 25
Altemately, Y is wherein each R" independently is H or Cj^alkyl, 30 and s is 1.
Altemately, Y is 011341 -6- R"R"N — , wherein G is S and each R" independently is H or C^alkyl.
Altemately, Y is
N
NR" , wherein R" is H or Cj^alkyl.
Représentative of the novel compounds of this invention are the following:(±)-10,11 -Dihydro-3-[2-(6-aminopyridin-2-y 1)-1 -ethoxy]-5H- 10 dibenzo[a,d]cycloheptene- 10-acetic acid; (±)-10,11 -Dihydro-3-[4-(pyridin-2-ylamino)-1 -butyl]-5H- dibenzo[a,d]cycloheptene-10-acetic acid; (±)-10,11 -Dihydro-3-[3-(4-ethoxypyridin-2-y lamino)-1 -propyloxy]-5H- dibenzo[a,d]cycloheptene-10-acetic acid; (S)-10,11 -Dihydro-3-[3-(py ridin-2-ylamino)-1 -propy loxy]-5H- 15 dibenzo[a,d]cycloheptene-10-acetic acid; (R) -10,11 -Dihydro-3-[3-(pyridin-2-y lamino)-1 -propyloxy]-5H-dibenzo[a,d]cycloheptene- 10-acetic acid; (±)-10,l l-Dihydro-3-[3-(3,4,5,6-tetrahydropyrimidin-2-ylamino)-l-propyloxy]-5H-dibenzo[a,d]cycloheptene-10-acetic acid; 20 (±)-10,l l-Dihydro-3-[2-[2-(ethylamino)thiazol-4-yl]-l-ethoxy]-5H- dibenzo[a,d]cycloheptene- 10-acetic acid; (±)-10,11 -Dihydro-3-[3-(isoquinoline-1 -y lamino)-1 -propy loxy]-5H-dibenzo[a,d]cycloheptene-10-acetic acid; (±)-10,11 -Dihydro-7-fluoro-3-[3-(pyridin-2-y lamino)-1 -propy loxy ]-5H-25 dibenzo[a,d]cycloheptene-10-acetic acid; (S) -10,11 -Dihydro-3-[3-(4-methylpyridin-2-ylamino)- 1-propyloxy ]-5H-dibenzo[a,d]cycloheptene-10-acetic acid; (S)-10,11 -Dihydro-3-[3-(4-ethoxypyridin-2-ylamino)- 1-propyloxy ]-5H-dibenzo[a,d]cycloheptene-10-acetic acid; 30 (±)-10,l l-Dihydro-6-methyl-3-[3-(pyridin-2-ylamino)-l-propyloxy]-5H- dibenzo[a,d]cycloheptene- 10-acetic acid; 011341 5 10 15 20 25 30 35 -7- (+)-10,11 -Dihydro-2-(dimethy lamino)methy !-7-fîuoro-3-[3-(pyridin-2-y lamino)-1 -propyloxy]-5H-dibenzo[a,d]cycloheptene-10-acetic acid; (S)-10,1 l-Dihydro-3-[3-[4-(2-propyloxy)pyridin-2-ylamino]-l-propyloxy]-5H-dibenzo[a,d]cycloheptene- 10-acetic acid; (S)-10,11 -Dihydro-3-[2-[6-(methy lamino)pyridin-2-yl]-1 -ethoxy]-5H-dibenzo[a,d]cycloheptene-10-acetic acid; (S)-10,11 -Dihydro-3-[3-[4-(dimethylamino)pyridin-2-ylamino]-1 -propyloxy]-5H-dibenzo[a,d]cycloheptene-10-acetic acid; (±)-10,11 -Dihydro-3-[3-[4-(ethyIthio)pyridin-2-y lamino]-1 -propy loxy]-5H-dibenzo[a,d]cycloheptene-l0-acetic acid; (S)-10,11 -Dihydro-3-[3-(4-chIoropyridin-2-ylamino)-1 -propyloxy ]-5H-dibenzo[a,d]cycloheptene-10-acetic acid; (±)-10,1 l-Dihydro-2-methyl-3-[3-(pyridin-2-ylamino)-1 -propy loxy]-5H-dibenzo[a,d]cycloheptene- 10-acetic acid; (S)-10,11 -Dihydro-3-[3-(4-aminopyridin-2-y lamino)-1 -propyloxy ]-5H-dibenzo[a,d]cycloheptene-10-acetic acid ; (±)-10,l l-Dihydro-3-[3-(4-methylpyridin-2-ylamino)-l-propyloxy]-dibenzo[b,f]oxepine-10-acetic acid; (±)-10,1 l-Dihydro-3-[2-[6-(methylamino)pyridin-2-yl]- 1-ethoxy]-dibenzo[b,f]oxepine-10-acetic acid; and (S)-10,11 -Dihydro-3-[3-(2-aminopyridin-4-yl)-1 -propy loxy ]-5H-dibenzo[a,d]cycloheptene- 10-acetic acid;or a pharmaceutically acceptable sait thereof.
In cases wherein the compounds of this invention may hâve one or more chiralcenters, unless specified, this invention includes each unique nonracemic compound whichmay be synthesized and resolved by conventional techniques. According to the présentinvention, the (S) configuration of the formula (I) compounds is preferred.
In cases in which compounds hâve unsaturated carbon-carbon double bonds, boththe cis (Z) and trans (E) isomers are within the scope of this invention. The meaning of anysubstituent at any one occurrence is independent of its meaning, or any other substituent’smeaning, at any other occurrence.
Also included in this invention are prodrugs of the compounds of this invention.Prodrugs are considered to be any covalently bonded carriers which release the activeparent drug according to formula (I) in vivo. Thus, in another aspect of this invention arenovel prodrugs, which are also intermediates in the préparation of formula (I) compounds,of formula (II): -8- 011341
(Π) wherein: 5 A is CH2 or O; R' is H, halo or Cj.^alkyl; R2 is H, Ci.^alkyl or CH2NR"R";X is O or CH2;
Yis
15 GisNR", S or O; R’ is H, Cj.galkyl, OCi.6alkyl, SCi.6alkyl, NR"R" or halo;each R" independently is H or Cj.galkyl; ands is 0, 1 or 2; or a pharmaceutically acceptable sait thereof. 2θ 1° yet another aspect of this invention are novel intermediates of formula (III): 011341 -9-
(III) 10 10 15 15 20 O- NR-—(CH2)2.3-XJj-
CO2C1<aikyl 20 25 25 wherein: A is CH2 or O; -~ R1 is H, halo or Cj.^alkyl; R2 is H, Cj-éalkyl or CH2NR"R"; X is O or CH2; R’ is H, Ci_6alkyl, OCi-6alkyl, SCi-6alkyl, NR"R" or halo; andeach R" independently is H or Cj.^alkyl; or a pharmaceutically acceptable sait thereof.
Abbreviations and symbols commonly used in the peptide and Chemical arts areused herein to describe the compounds of this invention. In general, the amino acidabbreviations follow the IUPAC-IUB Joint Commission on Biochemical Nomenclature asdescribed in Eur. J. Biochem., 158,9 (1984).
Cj.4alkyl as applied herein means an optionally substituted alkyl group of 1 to 4carbon atoms, and includes methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl and t-butyl.Cj.galkyl additionally includes pentyl, n-pentyl, isopentyl, neopentyl and hexyl and thesimple aliphatic isomers thereof. CQ.4alkyl and Cç^alkyl additionally indicates that noalkyl group need be présent (e.g., that a covalent bond is présent).
Any C^alkyl or Cj.g alkyl may be optionally substituted with the group Rx,which may be on any carbon atom that results in a stable structure and is available byconventional synthetic techniques. Suitable groups for Rx are C^alkyl, OR", SR",Cj^alkylsulfonyl, C1_4alkylsulfoxyl, -CN, N(R")2, CtyNiR"^, -NO2, -CF3, -CO2R",-CON(R")2, -COR”, -NR"C(O)R”, F, Cl, Br, I, or CF3S(O)r-, wherein r is 0, 1 or 2.
Halogen or halo means F, Cl, Br, and I.
Ar, or aryl, as applied herein, means phenyl or naphthyl, or phenyl or naphthylsubstituted by one to three substituents, such as those defined above for alkyl, especiallyC^alkyl, C[_4alkoxy, Ci.4alkthio, CF3, NH2, OH, F, Cl, Br or I.
Certain radical groups are abbreviated herein. t-Bu refers to the tertiary butylradical, Boc refers to the t-butyloxycarbonyl radical, Fmoc refers to thefluorenylmethoxycarbonyl radical, Ph refers to the phenyl radical, Cbz refers to thebenzyloxycarbonyl radical, Bn refers to the benzyl radical, Me refers to methyl, Et refers to 011341 - 10- ethyl, Ac refers to acetyl, Alk refers to Cj^alkyl, Nph refers to 1- or 2-naphthyl and cHexrefers to cyclohexyl. Tet refers to 5-tetrazolyl.
Certain reagents are abbreviated herein. DCC refers to dicyclohexylcarbodiimide,DMAP refers to dimethylaminopyridine, DIEA refers to diisopropylethyl amine, EDC 5 refers to l-(3-dimethylaminopropyl)-3-ethylcarbodiimide, hydrochloride. HOBt refers to1-hydroxybenzotriazole, THF refers to tetrahydrofuran, DIEA refers todiisopropylethylamine, DEAD refers to diethyl azodicarboxylate, PPhj refers totriphenylphosphine, DIAD refers to diisopropyl azodicarboxylate, DME refers todimethoxyethane, DMF refers to dimethylformamide, NBS refers to N-bromosuccinimide, 10 Pd/C refers to a palladium on carbon catalyst, PPA refers to polyphosphoric acid, DPPArefers to diphenylphosphoryl azide, BOP refers to benzotriazol-l-yloxy-tris(dimethyl-amino)phosphonium hexafluorophosphate, HF refers to hydrofluoric acid, TEA refers totriethylamine, TFA refers to trifluoroacetic acid, PCC refers to pyridinium chlorochromate.
The compounds of formula (I) are generally prepared by reacting a compound of 15 formula (IV) with a compound of formula (V):
20 wherein Rl, R A Y and A are as defined in formula (I), with any reactive functionalgroups protected, and lJ is OH or halo; and thereafter removing any protecting groups, and optionally forming apharmaceutically acceptable sait.
Suitably, certain compounds of formula (I) are prepared by reacting a compound offormula (IV) with a compound of formula (VI):
O-
25 011341 - 11 - wherein r7, R2, R’, R" and A are as defined in formula (I), with any reactivefunctional groups protected; and thereafter removing any protecting groups, and optionally forming apharmaceutically acceptable sait.
Suitably, the reaction between a compound of formual (IV) with a compound offormula (VI) is carried out in the presence of diethyl azodicarboxylate andtriphenylphosphine in an aprotic solvent.
Additionally, certain compounds of formula (I) are prepared by rêàcting acompound of formula (IV) with a compound of formula (VII): 10
wherein R\ R2, R" and A are as defined in formula (I), with any reactive15 functional groups protected; and thereafter removing any protecting groups, and optionally forming apharmaceutically acceptable sait.
Suitably, the reaction between a compound of formual (IV) with a compound offormula (VII) is carried out in the presence of diethyl azodicarboxylate and 20 triphenylphosphine in an aprotic solvent.
Compounds of the formula (I) are prepared by the methods described in Bondinell et al., PCT Publication No. WO 97/01540 (International Application No. PCT/US96/11108), published January 16,1997, the entire disclosure of which isincorporated herein by référencé. 25 Additionally, compounds of formula (I) are prepared by methods analogous to those described in the schemes that are detailed hereinafter. - 12- 011341
Scheme I
a) 10% Pd/C, HOAc; b) SOC12, toluene; c) A1C13, CH2CI,5
Scheme I details the préparation of an intermediate useful in the préparation offormula (I) compounds. 011341 - 13-
Scheme II
a) LiN(TMS)2, ethyl bromoacetate; b) Jones reagent, OsO4; c) H2, 10% Pd/C, HO AC;d) C2O2C12, DMF; e) A1C13, CH2C12, RT; f) H2, 10% Pd/C, HOAC
Scheme II also details the préparation of an intermediate useful in the préparationof formula (I) compounds. - 14- 011341
Sçheme III
011341 - 15 - (a) EtOAc/LiHMDS, THF; (b) H2, 10% Pd/C, conc. HCl, AcOH; (c) EtSH, AICI3, CH2CI2;(d) 2-[(3-hydroxy-l-propyl)amino]-4-nitropyridine-N-oxide, DEAD, (Ph)3P; (e) NaOEt,EtOH; (f) cyclohexene, 10% Pd/C, EtOH; (g) 1.0 N NaOH, EtOH; (h) HCl. 5 Scheme III details the préparation of a formula (I) compound. Reaction of „1-1 (which is a Scheme 1-3 compound) in an aldol-type reaction with the enolate of ethylacetate, which can be generated from ethyl acetate on exposure to an appropriate amidebase, for instance lithium diisopropylamide (LDA) or lithium bis(trimethylsilyl)amide(LiHMDS), gives „1-2. Frequently, THF is the solvent of choice for an aldol reaction, 10 although THF in the presence of various additives, for instance HMPA or TMEDA, is oftenused. Réduction of „1-2 to give „1-3 (which is a Scheme „-6 compound) can beaccomplished by hydrogenolysis over an appropriate catalyst, for example palladium métalon activated carbon (Pd/C), in an appropriate solvent, such as acetic acid, in the presence ofa minerai acid such as HCl. Altematively, this réduction can be accomplished by treatment 15 of „1-2 with triethylsilane in the presence of boron trifluoride etherate by the general method of Orfanopoulos and Smonou {Synth. Commun. 1988, 833). Removal of the methylether of „1-3 to give „1-4 can be accomplished with BBrç in an inert solvent, for exampleCH2CI2, or by reaction with ethanethiol and AICI3 in an inert solvent, preferably CH2CI2.Other useful methods for removal of a methyl ether are described in Greene, "Protective 20 Groups in Organic Synthesis" (published by John Wiley and Sons). Compound 4 of
Scheme 3 (III-4) is reacted with 2-[(3-hydroxy-l-propyl)amino]-4-nitropyridine-N-oxide ina Mitsunobu-type coupling reaction {Organic Reactions 1992,42, 335-656; Synthesis 1981,1-28) to afford III-5. The reaction is mediated by the complex formed between diethylazodicarboxylate and triphenylphosphine, and is conducted in an aprotic solvent, for 25 instance THF, CH2CI2, or DMF. Compound „1-5 is reacted with an alkali métal sait of anappropriate alcohol to afford „1-6. Suitable alkali metals include lithium, sodium,potassium, and césium, and the alcohol used for the displacement reaction is generally usedas the solvent. Methods for forming the alkali métal salts of alcohols are well-known tothose of skill in the art. The pyridine-N-oxide moiety of „1-6 is reduced to the 30 corresponding pyridine „1-7 under transfer hydrogénation conditions using a palladiumcatalyst, preferably palladium métal on activated carbon, in an inert solvent, for instancemethanol, éthanol, or 2-propanol. Cyclohexene, 1,4-cyclohexadiene, formic acid, and saltsof formic acid, such as potassium formate or ammonium formate, are commonly used asthe hydrogen transfer reagent in this type of reaction. The ethyl ester of III-7 is hydrolyzed 35 using aqueous base, for example, LiOH in aqueous THF or NaOH in aqueous methanol oréthanol, and the intermediate carboxylate sait is acidified with a suitable acid, for instanceTFA or HCl, to afford the carboxylic acid „1-8. Altematively, the intermediate 011341 - 16- carboxylate sait can be isolated, if desired. or a carboxylate sait of the free carboxylic acidcan be prepared by methods well-known to those of skill in the art.
Scheme IV
(a) NaH, 2-[N-(3-methanesulfonyloxy-l-propyl)-N-(ferr-butoxycarbonyl)amino]pyridine-N-oxide, DMSO; (b) TFA, CH2CI2; (c) see Scheme III. 10
Scheme IV describes an alternative method for the préparation of formula (I)compounds. Compound IV-1 is reacted with a base, preferably an alkali métal hydridesuch as sodium hydride or potassium hydride, in a polar, aprotic solvent, generally THF,DMF, DMSO, or mixtures thereof, to afford the corresponding alkali métal phenoxide. 15 Altematively, an alkali métal amide, for instance LDA, or the lithium, sodium, or potassium sait of hexamethyldisilazane, can be used for deprotonation. The intermediatephenoxide is generally not isolated, but is reacted in situ with an appropriate electrophile,for instance 2-[N-(3-methanesulfonyloxy-l-propyl)-N-(rerr-butoxycarbonyl)-amino]pyridine-N-oxide, to afford the coupled product IV-2. The rerr-butoxycarbonyl 20 protecting group in IV-2 is removed under acidic conditions, such as 4 M HCl in 1,4- 011341 - 17- dioxane or TFA in CH2CI2, to afford IV-3. Conditions for removal of the tert-butoxycarbony! protecting group are well-known to those of skili in the art, and severaiuseful methods are described in standard référencé volumes such as Greene "ProtectiveGroups in Organic Synthesis". IV-3 is subsequently converted to IV-4 following theprocedure outlined in Scheme III.
Scheme V
(a) PhOH, Cu, K2CO3; (b) sulfur, morpholine; (c) KOH, H2O, i-PrOH; (d) SOC12,benzene; (e) AICI3, CH2C12; (f) EtOAc, LiN(TMS)2, TMEDA, THF; (g) Et3SiH, BF3 ·OEt2, CH2C12; (h) H2, PdZC, EtOH; (i) BBr3, CH2C12. 15 Commercially available 2-fluoro-4-methoxyacetophenone (V-l) reacts with an alcohol, for example phénol, in the presence of copper métal and a suitable base, forinstance K2CO3, to afford the diaryl ether V-2. On treatment with sulfur and anappropriate primary or secondary amine, preferably morpholine, according to the general 011341 - 18- method of Harris (J. Med. Chem. 1982, 25, 855), V-2 is converted to V-3 in a classicalWillgerodt-Kindler reaction. The thioamide thus obtained is hydrolyzed to thecorresponding carboxylic acid V-4 by reaction with an alkali métal hydroxide, suitablyKOH, in an aqueous alcoholic solvent, such as aqueous MeOH, EtOH, or i-PrOH. 5 Carboxylic acid V-4 is converted to the corresponding acid chloride by reaction with eitherSOCl? or oxalyl chloride according to conditions well-known to those of skill in the art.Treatment of this acid chloride with an appropriate Friedel-Crafts catalyst, such as AICI3 orSnCl4, in an inert solvent, such as CH2CI2 or CSo, provides the cyclic ketone V-5.Altematively, acid V-4 can be converted directly to ketone V-5 under acidic conditions, for 10 example with polyphosphoric acid. Reaction of V-5 in an aldol - type reaction with theenolate of ethyl acetate, which can be generated from ethyl acetate on exposure to anappropriate amide base, for instance lithium diisopropylamide (LDA) or lithiumbis(trimethylsilyl)amide (LiHMDS), gives V-6. Frequently, THF is the solvent of choicefor an aldol reaction, although THF in the presence of various additives, for instance 15 HMPA or TMEDA, is often used. Réduction of V-6 to give V-7 can be accomplished bytreatment of V-6 with triethylsilane in the presence of boron trifluoride etherate by thegeneral method of Orphanopoulos and Smonu (Synth. Commun. 1988, 833). Any olefinicby-products that resuit from élimination of the alcohol are reduced by hydrogénation overan appropriate catalyst, for example palladium métal on activated carbon (Pd/C), in an 20 appropriate solvent, such as MeOH or EtOH. Altematively, the réduction of V-6 to giveV-7 can be accomplished by hydrogenolysis in the presence of a minerai acid such as HCl.Typically, this reaction is catalyzed by Pd/C, and is optimally conducted in acetic acid.Removal of the methyl ether of V-7 to give V-8 can be accomplished with BBr3 in an inertsolvent, for example CH2CI2, or by reaction with ethanethiol and AICI3 in an inert solvent, 25 preferably CH2CI2. Other useful methods for removal of a methyl ether are described inGreene, "Protective Groups in Organic Synthesis" (published by John Wiley and Sons). V-8 is subsequently converted to formula (I) compounds following the procedure outlinedin Scheme III.
Acid addition salts of the compounds are prepared in a standard manner in a 30 suitable solvent from the parent compound and an excess of an acid, such as hydrochloric,hydrobromic, hydrofluoric, sulfuric, phosphoric, acetic, trifluoroacetic, maleic, succinic ormethanesulfonic. Certain of the compounds form inner salts or zwitterions which may beacceptable. Cationic salts are prepared by treating the parent compound with an excess ofan alkaline reagent, such as a hydroxide, carbonate or alkoxide, containing the appropriate 35 cation; or with an appropriate organic amine. Cations such as Li+, Na+, K+, Ca++, Mg++and NH4+ are spécifie examples of cations présent in pharmaceutically acceptable salts. 011341 - 19-
This invention also provides a pharmaceutical composition which comprises acompound according to formula (I) and a pharmaceutically acceptable carrier.
Accordingly, the compounds of formula (I) may be used in the manufacture of amédicament. Pharmaceutical compositions of the compounds of formula (I) prepared as 5 hereinbefore described may be formulated as solutions or lyophilized powders for parentéral administration. Powders may be reconstituted by addition of a suitable diluentor other pharmaceutically acceptable carrier prior to use. The Iiquid formulation may be abuffered, isotonie, aqueous solution. Examples of suitable diluents are normal isotoniesaline solution, standard 5% dextrose in water or buffered sodium or ammonium acetate 10 solution. Such formulation is especially suitable for parentéral administration, but may alsobe used for oral administration or contained in a metered dose inhaler or nebulizer forinsufflation. It may be désirable to add excipients such as polyvinylpyrrolidone, gelatin,hydroxy cellulose, acacia, polyethylene glycol, mannitol, sodium chloride or sodiumcitrate. 15 Altemately, these compounds may be encapsulated, tableted or prepared in a émulsion or syrup for oral administration. Pharmaceutically acceptable solid or Iiquidcarriers may be added to enhance or stabilize the composition, or to facilitate préparation ofthe composition. Solid carriers include starch, lactose, calcium sulfate dihydrate, terra alba,magnésium stéarate or stearic acid, talc, pectin, acacia, agar or gelatin. Liquid carriers 20 include syrup, peanut oil, olive oil, saline and water. The carrier may also include a sustained release material such as glyceryl monostearate or glyceryl distearate, alone orwith a wax. The amount of solid carrier varies but, preferably, will be between about 20mg to about 1 g per dosage unit. The pharmaceutical préparations are made following theconventional techniques of pharmacy involving milling, mixing, granulating, and 25 compressing, when necessary, for tablet forms; or milling, mixing and fïlling for hard gelatin capsule forms. When a liquid carrier is used, the préparation will be in the form ofa syrup, élixir, émulsion or an aqueous or non-aqueous suspension. Such a liquidformulation may be administered directly p.o. or filled irito a soft gelatin capsule.
For rectal administration, the compounds of this invention may also be combined 30 with excipients such as cocoa butter, glycerin, gelatin or polyethylene glycols and moldedinto a suppository.
The compounds described herein are antagoniste of the vitronectin receptor, and areuseful for treating diseases wherein the underlying pathology is attributable to ligand or cellwhich interacts with the vitronectin receptor. For instance, these compounds are useful for 35 the treatment of diseases wherein loss of the bone matrix créâtes pathology. Thus, theinstant compounds are useful for the treatment of ostoeporosis, hyperparathyroidism,Paget’s disease, hypercalcemia of malignancy, osteolytic lésions produced by bone 011341 -20- metastasis, bone loss due to immobilization or sex hormone deficiency. The compounds ofthis invention are also believed to hâve utility as antitumor, anti-angiogenic,antiinflammatory and anti-metastatic agents, and be useful in the treatment ofatherosclerosis and restenosis. 5 The compound is administered either orally or parenterally to the patient, in a manner such that the concentration of drug is sufficient to inhibit bone résorption, or othersuch indication. The pharmaceutical composition containing the compound is administeredat an oral dose of between about 0.1 to about 50 mg/kg in a manner consistent with thecondition of the patient. Preferably the oral dose would be about 0.5 to about 20 mg/kg. 10 For acute therapy, parentéral administration is preferred. An intravenous infusion of thepeptide in 5% dextrose in water or normal saline, or a similar formulation with suitableexcipients, is most effective; although an intramuscular bolus injection is also useful.Typically, the parentéral dose will be about 0.01 to about 100 mg/kg; preferably between0.1 and 20 mg/kg. The compounds are administered one to four times daily at a level to 15 achieve a total daily dose of about 0.4 to about 400 mg/kg/day. The précisé level and method by which the compounds are administered is readily determined by one routinelyskilled in the art by comparing the blood level of the agent to the concentration required tohâve a therapeutic effect.
This invention further provides a method for treating osteoporosis or inhibiting 20 bone loss which comprises administering stepwise or in physical combination a compoundof formula (I) and other inhibitors of bone résorption, such as bisphosphonates (i.e.,allendronate), hormone replacement therapy, anti-estrogens, or calcitonin. In addition, thisinvention provides a method of treatment using a compound of this invention and ananabolic agent, such as the bone morphogenic protein, iproflavone, useful in the prévention 25 of bone loss and/or to increase bone mass.
Additionally, this invention provides a method of inhibiting tumor growth which comprises administering stepwise or in physical combination a compound of formula (I)and an antineoplastic agent. Compounds of the camptothecin analog class, such astopotecan, irinotecan and 9-aminocamptothecin, and platinum coordination complexes, 30 such as cisplatin, ormaplatin and tetraplatin, are well known groups of antineoplasticagents. Compounds of the camptothecin analog class are described in U.S. Patent Nos.5,004,758,4,604,463, 4,473,692, 4,545,880 4,342,776, 4,513,138, 4,399,276, EP PatentApplication Publication Nos .0 418 099 and 0 088 642, Wani, et al., J. Med. Chem., 1986,29, 2358, Wani, et al., J. Med. Chem., 1980, 23, 554, Wani, et al., J. Med. Chem., 1987, 30, 35 1774, and Nitta, et al., Proc. I4th International Congr. Chemotherapy., 1985, Anticancer
Section I, 28, the entire disclosure of each which is hereby incorporated by référencé. Theplatinum coordination complex, cisplatin, is available under the name Platinol® from 011341 -21 -
Bristol Myers-Squibb Corporation. Useful formulations forcisplatin are described in U.S.Patent Nos. 5,562,925 and 4,310,515, the entire disclosure of each wbich is herebyincorporated by référencé.
In the method of inhibiting tumor growth which comprises administering stepwise 5 or in physical combination a compound of formula (I) and an antineoplastic agent, theplatinum coordination compound, for example cisplatin, can be administered using slowintravenous infusion. The preferred carrier is a dextrose/saline solution containingmannitol. The dose schedule of the platinum coordination compound maybe on the basisof from about 1 to about 500 mg per square meter (mg/m^) of body surface area per course 10 of treatment. Infusions of the platinum coordiation compound may be given one to twotimes weekly, and the weekly treatments may be repeated several times. Using acompound of the camptothecin analog class in a parentéral administration, the course oftherapy generally employed is from about 0.1 to about 300.0 mg/m^ of body surface areaper day for about five consecutive days. Most preferably, the course of therapy employed 15 for topotecan is from about 1.0 to about 2.0 mg/m^ of body surface area per daÿ for aboutfive consecutive days. Preferably, the course of therapy is repeated at least once at about aseven day to about a twenty-eight day interval.
The pharmaceutical composition may be formulated with both the compound offormula (I) and the antineoplastic agent in the same container, but formualtion in different 20 containers is preferred. When both agents are provided in solution form, they can be contained in an infusion/injection System for simultaneous administration or in a tandemarrangement.
For convenient administration of the compound of formula (I) and theantineoplastic agent at the same or different times, a kit is prepared, comprising, in a single 25 container, such as a box, carton or other container, individual bottles, bags, vials or othercontainers each having an effective amount of the compound of formula (I) for parentéraladministration, as described above, and an effective amount of the antineoplastic agent forparentéral administration, as described above. Such kit can comprise, for example, bothpharmaceutical agents in separate containers or the same container, optionally as 30 lyophilized plugs, and containers of solutions for reconstitution. A variation of this is toinclude the solution for reconstitution and the lyophilized plug in two chambers of a singlecontainer, which can be caused to admix prior to use. With such an arrangement, theantineoplastic agent and the compound of this invention may be packaged separately, as intwo containers, or lyophilized together as a powder and provided in a single container. 35 When both agents are provided in solution form, they can be contained in an infusion/injection System for simultaneous administration or in a tandem arrangement. Forexample, the compound of formula (I) may be in an i.v. injectable form, or infusion bag 011341 -22- linked in sériés, via tubing, to the antineoplastic agent in a second infusion bag. Using sucha System, a patient can receive an initial bolus-type injection or infusion of the compoundof formula (I) followed by an infusion of the antineoplastic agent.
The compounds may be tested in one of several biological assays to détermine the 5 concentration of compound which is required to hâve a given pharmacological effect.
Inhibition of vitronectin binding
Solid-Phase (^H]-SK&F-107260 Binding to ανβ^: Human placenta or human plateletανβ3 (0.1-0.3 mg/mL) in buffer T (containing 2 mM CaCl2 and 1% octylglucoside) was 10 diluted with buffer T containing 1 mM CaCl2, 1 mM MnCl2, 1 mM MgCl2 (buffer A) and0.05% NaN3, and then immediately added to 96-well ELISA plates (Corning, New York,NY) at 0.1 mL per well. 0.1 - 0.2 pg of ανβ3 was added per well. The plates wereincubated ovemight at 4°C. At the time of the experiment, the wells were washed oncewith buffer A and were incubated with 0.1 mL of 3.5% bovine sérum albumin in the same 15 buffer for 1 hr at room température. Following incubation the wells were aspiratedcompletely and washed twice with 0.2 mL buffer A.
Compounds were dissolved in 100% DMSO to give a 2 mM stock solution, whichwas diluted with binding buffer (15 mM Tris-HCl (pH 7.4), 100 mM NaCl, 1 mM CaCl2, 1mM MnCl2, 1 mM MgCl2) to a final compound concentration of 100 μΜ. This solution is 20 then diluted to the required final compound concentration. Various concentrations of unlabeled antagoniste (0.001 -100 μΜ) were added to the wells in triplicates, followed bythe addition of 5.0 nM of [3H]-SK&F-107260 (65 - 86 Ci/mmol).
The plates were incubated for 1 hr at room température. Following incubation thewells were aspirated completely and washed once with 0.2 mL of ice cold buffer A in a 25 well-to-well fashion. The receptors were solubilized with 0.1 mL of 1% SDS and thebound [3H]-SK&F-107260 was determined by liquid scintillation counting with theaddition of 3 mL Ready Safe in a Beckman LS Liquid Scintillation Counter, with 40%efficiency. Nonspecific binding of [3H]-SK&F-107260 was determined in the presence of2 μΜ S K&F-107260 and was consistently less than 1% of total radioligand input. The 30 IC50 (concentration of the antagonist to inhibit 50% binding of [3H]-SK&F-107260) was determined by a nonlinear, least squares curve-fitting routine, which was modified from theLUNDON-2 program. The Kj (dissociation constant of the antagonist) was calculatedaccording to the équation: Kj = IC50/( 1 + L/K<i), where L and K4 were the concentrationand the dissociation constant of [3H]-SK&F-107260, respectively. 35 Compounds of the présent invention inhibit vitronectin binding to SK&F 107260 in the concentration range of about 2.5 to about 0.001 micomolar. 011341 - 23 -
Compounds of this invention are also tested for in vitro and in vivo bone résorptionin assays standard in the art for evaluating inhibition of bone formation, such as the pitformation assay disclosed in EP 528 587, which may also be performed using humanosteoclasts in place of rat osteoclasts, and the ovarectomized rat mode!, described by 5 Wronski et al., Cells and Materials 1991, Sup. 1,69-74.
Vascular smooth muscle cell migration assay
Rat or human aortic smooth muscle cells were used. The cell migration wasmonitored in a Transwell cell cùlture chamber by using a polycarbonate membrane with 10 pores of 8 um (Costar). The lower surface of the filter was coated with vitronectin. Cellswere suspended in DMEM supplemented with 0.2% bovine sérum albumin at aconcentration of 2.5 - 5.0 x 10^ cells/mL, and were pretreated with test compound atvarious concentrations for 20 min at 20°C. The solvent alone was used as control. 0.2 mLof the cell suspension was placed in the upper compartment of the chamber. The lower 15 compartment contained 0.6 mL of DMEM supplemented with 0.2% bovine sérum albumin.Incubation was carried out at 37°C in an atmosphère of 95% air/5% CO2 for 24 hr. Afterincubation, the non-migrated cells on the upper surface of the filter were removed by gentlescraping. The filter was then fixed in methanol and stained with 10% Giemsa stain.
Migration was measured either by a) counting the number of cells that had migrated to the 20 lower surface of the filter or by b) extracting the stained cells with 10% acetic acidfollowed by determining the absorbance at 600 nM.
Thyroparathyroidectomized rat model
Each experimental group consists of 5-6 adult male Sprague-Dawley rats (250-400g body 25 weight). The rats are thyroparathyroidectomized (by the vendor, Taconic Farms) 7 days prior touse. Ail rats receive a replacement dose of thyroxine every 3 days. On receipt of the rats,circulating ionized calcium levels are measured in whole blood immediately after it has beenwithdrawn by tail venipuncture into heparinized tubes. Rats are included if the ionized Ca level(measured with a Ciba-Coming model 634 calcium pH analyzer) is <1.2 mM/L. Each rat is fitted 30 with an indwelling venous and arterial cathéter for the delivery of test material and for blood sampling respectively. Thé rats are then put on a diet of calcium-free chow and deionized water.Baseline Ca levels are measured and each rat is administered either control vehicle or humanparathyroid hormone 1-34 peptide (hPTHl-34, dose 1.25 ug/kg/h in saline/0.1% bovine sérumalbumin, Bachem, Ca) or a mixture of hPTHl-34 and test material, by continuous intravenous 35 infusion via the venous cathéter using an extemal syringe pump. The calcémie response of eachrat is measured at two-hourly intervals during the infusion period of 6-8 hours. -24-
Human osteoclast résorption and adhesion assays
Pit résorption and adhesion assays hâve been developed and standardized using normal human osteoclasts derived from osteoclastoma tissue. Assay 1 was developed forthe measurement of osteoclast pit volumes by laser confocal microscopy. Assay 2 was 5 developed as a higher throughput screen in which collagen fragments (released duringrésorption) are measured by competitve ELISA.
Assay 1 (using laser confocal microscopy) • Aliquots of human osteoclastoma-derived cell suspensions are removed from liquid10 nitrogen strorage, warmed rapidly at 37°C and washed xl in RPMI-1640 medium by centrifugation (lOOOrpm, 5 mins at 4°C). • The medium is aspirated and replaced with murine anti-HLA-DR antibody thendiluted 1:3 in RPMI-1640 medium. The suspension is incubated for 30 mins on ice andmixed frequently. 15 · The cells are washed x2 with cold RPMI-1640 followed by centrifugation (1000 rpm, 5 mins at 4°C) and the cells are then transferred to a stérile 15 ml centrifuge tube.The number of mononuclear cells are enumerated in an improved Neubauer countingchamber.
• Sufficient magnetic beads (5 / mononuclear cell), coated with goat anti-mouse IgG 20 (Dynal, Great Neck, NY) are removed from their stock bottle and placed into 5 ml of fresh medium (this washes away the toxic azide preservative). The medium is removedby immobilizing the beads on a magnet and is replaced with fresh medium. • The beads are mixed with the cells and the suspension is incubated for 30 mins onice. The suspension is mixed frequently. 25 · The bead-coated cells are immobilized on a magnet and the remaining cells (osteoclast-rich fraction) are decanted into a stérile 50 ml centrifuge tube. • Fresh medium is added to the bead-coated cells to dislodge any trapped osteoclasts.This wash process is repeated xlO. The bead-coated cells are discarded. • The viable osteoclasts are enumerated in a counting chamber, using fluorescein 30 diacetate to label live cells. A large-bore disposable plastic pasteur pipet is used to add the sample to the chamber. • The osteoclasts are pelleted by centrifugation and the density adjusted to theappropriate number in EMEM medium (the number of osteoclasts is variable fromtumor to tumor), supplemented with 10% fêtai calf sérum and 1.7g/liter of sodium 35 bicarbonate. • 3ml aliquots of the cell suspension (per compound treatment) are decanted into15ml centrifuge tubes. The cells are pelleted by centrifugation. 011341 -25- • Το each tube, 3ml of the appropriate compound treatment are added (diluted to 50uM in the EMEM medium). Also included are appropriate vehicle Controls, a positivecontrol (anti-vitronectin receptor murine monoclonal antibody [87MEM1] diluted to100 ug/ml) and an isotype control (IgG,a diluted to 100 ug/ml). The samples are 5 incubated at 37°C for 30 mins. • 0.5ml aliquots of the cells are seeded onto stérile dentine slices in a 48-well plateand incubated at 37°C for 2 hours. Each treatment is screened in quadruplicate. • The slices are washed in six changes of warm PBS (10 ml / well in â 6-well plate)and then placed into fresh medium containing the compound treatment or control 10 samples. The samples are incubated at 37°C for 48 hours.
Tartrate résistant acid phosphatase (TRAP) procedure (sélective stainfor cells oftheosteoclast lineage) • The bone slices containing the attached osteoclasts are washed in phosphate 15 buffered saline and fixed in 2% gluteraldehyde (in 0.2M sodium cacodylate) for 5 mins. • They are then washed in water and are incubated for 4 minutes in TRAP buffer at37°C (0.5 mg/ml naphthol AS-BI phosphate dissolved in Ν,Ν-dimethylformamide andmixed with 0.25 M citrate buffer (pH 4.5), containing 10 mM sodium tartrate. • Following a wash in cold water the slices are immersed in cold acetate buffer (0.120 M, pH 6.2) containing 1 mg/ml fast red gamet and incubated at 4°C for 4 minutes. • Excess buffer is aspirated, and the slices are air dried following a wash in water. • The TRAP positive osteoclasts (brick red/ purple precipitate) are enumerated bybright-field microscopy and are then removed from the surface of the dentine bysonication. 25 · Pit volumes are determined using the Nikon/Lasertec ILM21W confocal microscope.
Assay 2 (using an ELISA readout)
The human osteoclasts are enriched and prepared for compound screening as30 described in the initial 9 steps of Assay 1. For clarity, these steps are repeated hereinbelow. • Aliquots of human osteoclastoma-derived cell suspensions are removed from liquidnitrogen strorage, wàrmed rapidly at 37°C and washed xl in RPMI-1640 medium bycentrifugation (lOOOrpm, 5 mins at 4°C). • The medium is aspirated and replaced with murine anti-HLA-DR antibody then35 diluted 1:3 in RPMI-1640 medium. The suspension is incubated for 30 mins on ice and mixed frequently. -26- 011341 • The cells are washed x2 with cold RPMI-1640 followed by centrifugation ( 1000rpm, 5 mins at 4°C) and the cells are then transferred to a stérile 15 ml centrifuge tube.The number of mononuclear cells are enumerated in an improved Neubauer countingchamber.
5 · Sufficient magnetic beads (5 / mononuclear cell), coated with goat anti-mouse IgG (Dynal, Great Neck, NY) are removed from their stock bottle and placed into 5 ml offresh medium (this washes away the toxic azide preservative). The medium is removedby immobilizing the beads on a magnet and is replaced with fresh medium. • The beads are mixed with the cells and the suspension is incubated for 30 mins on 10 ice. The suspension is mixed frequently. • The bead-coated cells are immobilized on a magnet and the remaining cells(osteoclast-rich fraction) are decanted into a stérile 50 ml centrifuge tube. • Fresh medium is added to the bead-coated cells to dislodge any trapped osteoclasts.This wash process is repeated xlO. The bead-coated cells are discarded. 15 · The viable osteoclasts are enumerated in a counting chamber, using flûorescein diacetate to label live cells. A large-bore disposable plastic pasteur pipet is used to addthe sample to the chamber. • The osteoclasts are pelleted by centrifugation and the density adjusted to theappropriate number in EMEM medium (the number of osteoclasts is variable from 20 tumor to tumor), supplemented with 10% fêtai calf sérum and 1,7g/liter of sodiumbicarbonate.
In contrast to the method desribed above in Assay 1, the compounds are screened at4 doses to obtain an ICM, as outlined below: • The osteoclast préparations are preincubated for 30 minutes at 37°C with test 25 compound (4 doses) or Controls. • They are then seeded onto bovine cortical bone slices in wells of a 48-well tissueculture plate and are incubated for a further 2 hours at 37°C. • The bone slices are washed in six changes of warm phosphate buffered saline(PBS), to remove non-adherent cells, and are then retumed to wells of a 48 well plate 30 containing fresh compound or Controls. • The tissue culture plate is then incubated for 48 hours at 37°C.
• The supematants from each well are aspirated into individual tubes and arescreened in a compétitive ELISA that detects the c-telopeptide of type I collagen whichis released during the résorption process. This is a commercially available ELISA 35 (Osteometer, Denmark) that contains a rabbit antibody that specifically reacts with an 8-amino acid sequence (Glu-Lys-Ala-His- Asp-Gly-Gly-Arg) that is présent in thecarboxy-terminal telopeptide of the al-chain of type I collagen. The results are -27- 011341 expressed as % inhibition of résorption compared to a vehicle contre!.
Human osteoclast adhesion assay
The human osteoclasts are enriched and prepared for compound screening as 5 described above in the inital 9 steps of Assay 1. For clarity, these steps are repeatedhereinbelow. • Aliquots of human osteoclastoma-derived cell suspensions are removed from liquidnitrogen strorage, warmed rapidly at 37°C and washed x 1 in RPMI-1640 medium bycentrifugation (lOOOrpm, 5 mins at 4°C). 10 · The medium is aspirated and replaced with murine anti-HLA-DR antibody then diluted 1:3 in RPMI-1640 medium. The suspension is incubated for 30 mins on ice andmixed frequently. • The cells are washed x2 with cold RPMI-1640 followed by centrifugation ( 1000rpm, 5 mins at 4°C) and the cells are then transferred to a stérile 15 ml centrifuge tube. 15 The number of mononuclear cells are enumerated in an improved Neubauer countingchamber. • Sufficient magnetic beads (5 / mononuclear cell), coated with goat anti-mouse IgG(Dynal, Great Neck, NY) are removed from their stock bottle and placed into 5 ml offresh medium (this washes away the toxic azide preservative). The medium is removed 20 by immobilizing the beads on a magnet and is replaced with fresh medium. • The beads are mixed with the cells and the suspension is incubated for 30 mins onice. The suspension is mixed frequently. • The bead-coated cells are immobilized on a magnet and the remaining cells(osteoclast-rich fraction) are decanted into a stérile 50 ml centrifuge tube. 25 · Fresh medium is added to the bead-coated cells to dislodge any trapped osteoclasts.
This wash process is repeated xlO. The bead-coated cells are discarded. • The viable osteoclasts are enumerated in a counting chamber, using fluoresceindiacetate to label live cells. A large-bore disposable plastic pasteur pipet is used to addthe sample to the chamber. 30 · The osteoclasts are pelleted by centrifugation and the density adjusted to the appropriate number in EMEM medium (the number of osteoclasts is variable fromtumor to tumor), supplemented with 10% fêtai calf sérum and 1.7g/liter of sodiumbicarbonate. • Osteoclastoma-derived osteoclasts are preincubated with compound (4 doses) or 35 Controls at 37°C for 30 minutes. • The cells are then seeded onto osteopontin-coated slides (human or rat osteopontin,2:5ug/ml) and incubated for 2 hours at 37°C. 011341 -28- • Non adhèrent cells are removed by washing the slides vigorously in phosphatebuffered saline and the cells remaining on the slides are fixed in acetone. • The osteoclasts are stained for tartrate-resistant acid phosphatase (TRAP), asélective marker for cells of this phenotype (see steps 15 -17), and are enumerated by 5 light microscopy. The results are expressed as % inhibition of adhesion compared to avehicle control.
Cell Adhesion AssayCells and Cell Culture
Human embryonic kidney cells ( HEK293 cells) were obtained from ATCC10 (Catalog No. CRL 1573). Cells were grown in Earl’s minimal essential medium (EMEM) medium containing Earl’s salts, 10% fêtai bovine sérum, 1% glutamine and 1% Penicillin-Steptomycin.
Constructs and Transfections A 3.2 kb EcoRI-Kpnl fragment of the av subunit and a 2.4 kb Xbal- Xhol fragment15 of the β3 subunit were inserted into the EcoRI - EcoRV cloning sites of the pCDN vector (Aiyar et al., 1994 ) which contains a CMV promoter and a G418 selectable marker byblunt end ligation. For stable expression, 80 x 10 θ HEK 293 cells were electrotransformedwith αν+β3 constructs (20 gg DNA of each subunit) using a Gene Puiser (Hensley et al.,1994 ) and plated in 100 mm plates (5x10^ cells/plate). After 48 hr, the growth medium 20 was supplemented with 450 gg/mL Geneticin (G418 Sulfate, GIBCO-BRL, Bethesda, MD).The cells were maintained in sélection medium until the colonies were large enough to beassayed.
Immunocytochemical analysis of transfected cells
To détermine whether the HEK 293 transfectants expressed the vitronectin 25 receptor, the cells were immobilized on glass microscope slides by centrifugation, fixed inacetone for 2 min at room température and air dried. Spécifie reactivity with 23C6, amonoclonal antibody spécifie for the ανβ3 complex was demonstrated using a standardindirect immunofluorescence method. 30 Cell Adhesion Studies
Corning 96-well ELISA plates were precoated ovemight at 4°C with 0.1 mL ofhuman vitronectin (0.2 gg/mL in RPMI medium). At the time of the experiment, the plateswere washed once with RPMI medium and blocked with 3.5% BSA in RPMI medium for 1hr at room température. Transfected 293 cells were resuspended in RPMI medium, 35 supplemented with 20 mM Hepes, pH 7.4 and 0.1 % BSA at a density of 0.5 x 10^ cells/mL. 011341 -29- 0.1 mL of cell suspension was added to each well and incubated for 1 hr at 37°C, in thepresence or absence of various ανβ3 antagonists. Following incubation, 0.025 mL of a10% formaldéhyde solution, pH 7.4, was added and the cells were fixed at roomtempérature for 10 min. The plates were washed 3 times with 0.2 mL of RPMI medium 5 and the adhèrent cells were stained with 0.1 mL of 0.5% toluidine blue for 20 min at roomtempérature. Excess stain was removed by extensive washing with deionized water. Thetoluidine blue incorporated into cells was eluted by the addition of 0.1 mL of 50% éthanolcontaining 50 mM HCl. Cell adhesion was quantitated at an optical density of 600 nm on amicrotiter plate reader (Titertek Multiskan MC, Sterling, VA). 10
Solid-Phase ccv3g Binding Assay:
The vitronectin receptor ανβ5 was purified from human placenta. Receptorpréparation was diluted with 50 mM Tris-HCl, pH 7.5, 100 mM NaCl, 1 mM CaCl2, 1 mMMnCl2, 1 mM MgCb (buffer A) and was immediately added to 96-well ELISA plates at 0.1 15 ml per well. 0.1-0.2 pg of ανβ3 was added per well. The plates were incubated ovemight at4°C. At the time of the experiment, the wells were washed once with buffer A and wereincubated with 0.1 ml of 3.5% bovine sérum albumin in the same buffer for 1 hr at roomtempérature. Following incubation the wells were aspirated completely and washed twicewith 0.2 ml buffer A. 20 In a [3H]-SK&F-107260 compétition assay, various concentrations of unlabeled
antagonists (0.001-100 μΜ) were added to the wells, followed by the addition of 5.0 nM of[3H]-SK&F-107260. The plates were incubated for 1 hr at room température. Followingincubation the wells were aspirated completely and washed once with 0.2 ml of ice coldbuffer A in a well-to-well fashion. The receptors were solubilized with 0.1 ml of 1% SDS 25 and the bound [3H]-SK&F-107260 was determined by liquid scintillation counting with theaddition of 3 ml Ready Safe in a Beckman LS 6800 Liquid Scintillation Counter, with 40%efficiency. Nonspecific binding of [3H]-SK&F-107260 was determined in the presence of 2μΜ SK&F-107260 and was consistently less than 1% of total radioligand input. The IC50(concentration of the antagonist to inhibit 50% binding of [3H]-SK&F-107260) was 30 determined by a nonlinear, least squares curve-fitting routine, which was modified from theLUNDON-2 program. The K; (dissociation constant of the antagonist) was calculatedaccording to Cheng and Prusoff équation: Kj = IC50/ (1 + L/Kd), where L and K<j were theconcentration and the dissociation constant of [3H]-SK&F-107260, respectively. 011341 - 30-
Inhibition of RGD-mediated GPIIb-IIIa binding
Purification of GPIIb-IIIa
Ten units of outdated, washed human platelets (obtained from Red Cross) were5 lyzed by gentle stirring in 3% octylglucoside, 20 mM Tris-HCl, pH 7.4, 140 mM NaCl, 2 mM CaCl2 at 4°C for 2 h. The lysate was centrifuged at 100,000g for 1 h. The supematantobtained was applied to a 5 mL lentil lectin sepharose 4B column (E.Y. Labs)preequilibrated with 20 mM Tris-HCl, pH 7.4, 100 mM NaCl, 2 mM CaCl2, 1%octylglucoside (buffer A). After 2 h incubation, the column was washed with 50 mL cold 10 buffer A. The lectin-retained GPIIb-IIIa was eluted with buffer A containing 10% dextrose.Ail procedures were performed at 4°C. The GPIIb-IIIa obtained was >95% pure as shownby SDS polyacrylamide gel electrophoresis.
Incorporation of GPIIb-IIIa in Liposomes. 15 A mixture of phosphatidylserine (70%) and phosphatidylcholine (30%) (Avanti
Polar Lipids) were dried to the walls of a glass tube under a stream of nitrogen. PurifiedGPIIb-IIIa was diluted to a final concentration of 0.5 mg/mL and mixed with thephospholipids in a protein:phospholipid ratio of 1:3 (w:w). The mixture was resuspendedand sonicated in a bath sonicator for 5 min. The mixture was then dialyzed ovemight using 20 12,000-14,000 molecular weight cutoff dialysis tubing against a 1000-fold excess of 50 mM Tris-HCl, pH 7.4, 100 mM NaCl, 2 mM CaC12 (with 2 changes). The GPIIb-IIIa-containing liposomes wee centrifuged at 12,000g for 15 min and resuspended in the dialysisbuffer at a final protein concentration of approximately 1 mg/mL. The liposomes werestored at -70C until needed. 25
Compétitive Binding to GPIIb-IIIa
The binding to the fibrinogen receptor (GPIIb-IIIa) was assayed by an indirectcompétitive binding method using pH]-SK&F-107260 as an RGD-type ligand. Thebinding assay was performed in a 96-weIl filtration plate assembly (Millipore Corporation, 30 Bedford, MA) using 0.22 um hydrophilic durapore membranes. The wells were precoatedwith 0.2 mL of 10 pg/mL polylysine (Sigma Chemical Co., St. Louis, MO.) at roomtempérature for 1 h to block nonspecific binding. Various concentrations of unlabeledbenzazepines were added to the wells in quadruplicate. pH]-SK&F-107260 was applied toeach well at a final concentration of 4.5 nM, followed by the addition of 1 pg of the purified 35 platelet GPIIb-IIIa-containing liposomes. The mixtures were incubated for 1 h at roomtempérature. The GPIIb-IIIa-bound [3HJ-SK&F-107260 was seperated from the unboundby filtration using a Millipore filtration manifold, followed by washing with ice-cold buffer 011341 -31 - (2 times, each 0.2 mL). Bound radioactivity remaining on the filters was counted in 1.5 mLReady Solve (Beckman Instruments, Fullerton, CA) in a Beckman Liquid ScintillationCounter (Model LS6800), with 40% efficiency. Nonspecific binding was determined in thepresence of 2 μΜ unlabeled SK&F-107260 and was consistently less than 0.14% of the 5 total radioactivity added to the samples. Ail data points are the mean of quadruplicatedéterminations.
Compétition binding data were analyzed by a nonlinear least-squares curve fittingprocedure. This method provides the IC50 of the antagonists (concentration of theantagonist which inhibits spécifie binding of [^H]-SK&F-107260 by 50% at equilibrium). 10 The IC50 is related to the equilibrium dissociation constant (Ki) of the antagonist based onthe Cheng and Prusoff équation: Ki = IC50/( 1+L/Kd), where L is the concentration of [3H]-SK&F-107260 used in the compétitive binding assay (4.5 nM), and Kd is the dissociationconstant of [3HJ-SK&F-107260 which is 4.5 nM as determined by Scatchard analysis.
Preferred compounds of this invention hâve an affinity for the vitronectin receptor 15 relative to the fibrinogen receptor of greater than 10:1. Most preferred compounds hâve aratio of activity of greater than 100:1.
The efficacy of the compounds of formula (I) alone or in combination with anantineoplastic agent may be determined using several transplantable mouse tumor models. 20 See U. S. Patent Nos. 5,004,758 and 5,633,016 for details of these models
The examples which follow are intended in no way to limit the scope of thisinvention, but are provided to illustrate how to make and use the compounds of thisinvention. Many other embodiments will be readily apparent to those skilled in the art. 25 011341 - 32-
EXAMPLES
General 5 Proton nuclear magnetic résonance ( NMR) spectra were recorded at either 250 or 400 MHz. Chemical shifts are reported in parts per million (δ) downfield from theinternai standard tetramethylsilane (TMS). Abbreviations for NMR data are as follows:s=singlet, d=doublet, t=triplet, q=quartet, m=multiplet, dd=doublet of doublets, dt=doubletof triplets, app=apparent, br=broad. J indicates the NMR coupling constant measured in 10 Hertz. CDCI3 is deuteriochloroform, DMSO-d6 is hexadeuteriodimethylsulfoxide, andCD3OD is tetradeuteriomethanol. Infrared (IR) spectra were recorded in transmissionmode, and band positions are reported in inverse wavenumbers (cm'l). Mass spectra wereobtained using electrospray (ES) or FAB ionization techniques. Elemental analyses wereperformed either in-house or by Quantitative Technologies Inc., Whitehouse, NJ. Melting 15 points were taken on a Thomas-Hoover melting point apparatus and are uncorrected. Ail températures are reported in degrees Celsius. Analtech Silica Gel GF and E. Merck SilicaGel 60 F-254 thin layer plates were used for thin layer chromatography. Both flash andgravity chromatography were carried out on E. Merck Kieselgel 60 (230-400 mesh) silicagel. Analytical and préparative HPLC were carried out on Rainin or Beckman 20 chromatographs. ODS refers to an octadecylsilyl derivatized silica gel chromatographiesupport. 5 μ Apex-ODS indicates an octadecylsilyl derivatized silica gel chromatographiesupport having a nominal particle size of 5 μ, made by Jones Chromatography, Littleton,Colorado. YMC ODS-AQ® is an ODS chromatographie support and is a registeredtrademark of YMC Co. Ltd., Kyoto, Japan. PRP-1 ® is a polymeric (styrene- 25 divinylbenzene) chromatographie support, and is a registered trademark of Hamilton Co.,Reno, Nevada. Celite® is a filter aid composed of acid-washed diatomaceous silica, and isa registered trademark of Manville Corp., Denver, Colorado.
Ethyl (±)-10,l l-dihydro-3-methoxy-5H-dibenzo[a,d]cycloheptene-10-acetate, ethyl(±)-10,l l-dihydro-3-hydroxy-5H-dibenzo[a,d]cycloheptene-10-acetate, and ethyl (±)- 30 10,1 l-dihydro-3-(trifluoromethanesulfonyloxy)-5H-dibenzo[a,d]cycloheptene-10-acetate were prepared according to WO 9701540-Al. 2-[2-(4-Methoxybenzylamino)pyridin-6-yl]ethanol was prepared according to WO 95/32710. 6-Methoxy-l-indanone was preparedby the method of House and Hudson (7. Org. Chem. 1970, 35, 647). -33- 011341
Prono ro t « λπA I V|>UIUHVII
Préparation of 2-f(3-hvdroxv-l-propyl)amino1pvridine-N-oxide 5 a) 2-[(3-Hydroxy-l-propyl)amino]pyridine-N-oxide A mixture of 2-chloropyridine-N-oxide hydrochloride (16.6 g, 0.1 mole), 3-amino-1-propanol (15.3 mL, 0.2 mole), NaHCO3 (42 g, 0.5 mole), and rerr-amyl alcohol (100 mL)was heated to reflux. After 21 hr, the reaction was cooled, diluted with CH2CI2 (300 mL),and suction filtered to remove insoluble materials. The filtrate was concentrated and10 reconcentrated from toluene to leave a yellow oil. Silica gel chromatography (20%MeOH/CHCl3) gave the title compound (15.62 g, 93%) as a yellow solid: TLC (20%MeOH/CHCl3) Rf 0.48; ’H NMR (250, CDCI3) δ 8.07 (dd, J = 6.6, 1.2 Hz, 1 H), 7.34 (br t,1 H), 7.10 - 7.30 (m, l H), 6.64 (dd, J = 8.5, 1.4 Hz, 1 H), 6.40 - 6.60 (m, 1 H), 4.49 (br s, 1H), 3.65 - 3.90 (m, 2 H), 3.35 - 3.60 (m, 2 H), 1.75 - 2.00 (m, 2 H); MS (ES) m/e 169 (M+ 15 H)+.
Préparation 2
Préparation of 2-i(3-hvdroxy-l-propvl)aminol-4-nitropyridine-N-oxide 20 a) 2-Chloro-4-nitropyridine-N-oxide A solution of conc. H2SO4 (30 mL) and fuming HNO3 (54 mL) was addeddropwise at 0°C to a solution of 2-chloropyridine-N-oxide hydrochloride (15.2 g, 91.56mmole) in conc. H2SO4 (30 mL). The reaction mixture was heated at 90°C for 1 hr, then25 was cooled to RT and poured onto ice (500 g). The reaction mixture was kept at RTovemight, then was cooled in an ice bath, and 50% NaOH was added slowly to give aprecipitate. This was collected and dried to give the title compound (5.88 g, 37 %) as apale yellow solid: !H NMR (400 MHz, CDCI3) δ 8.42 -8.37 (m, 2 H), 8.06 -8.04 (m, 1 H). 30 a) 2-[(3-Hydroxy-l-propyl)amino]-4-nitropyridine-N-oxide
According to the procedure of Préparation 1, except substituting 2-chloro-4-nitropyridine-N-oxide for the 2-chloropyridine-N-oxide hydrochloride, the title compoundwas obtained as yellow powder following silica gel chromatography (1:9 MeOH/C^C^)·Recrystallization from MeOH/CH2Cl2/Et2O gave the title compound: MS (ES) 214.135 . (M+H)+. -34- 011341
Préparation 3
Préparation of 2-i(3-hydroxv-l-propvl)amino1-4-methylpvridine-N-oxide 5 a) 2-Chloro-4-methylpyridine
Sodium nitrite (13.88 g, 200 mmole) was added slowly at 0°C to a solution of 2-amino-4-picoline (15.0 g, 139 mmole) in conc. HCl (200 mL). The reaction mixture wasallowed to warm to RT and was stirred for 16 hr, then was poured onto ice (500 g). The pHwas adjusted to 8.0 with conc. NH4OH, and the mixture was extracted with ether (3 x 300 10 mL). The combined ether layers were washed sequentially with H2O (2 x 200 mL) and brine (200 mL). Drying (MgSC>4) and concentration gave the title compound (10.3 g, 58%)as a faintly yellow oil: MS (ES) m/e 127.8 (M + H)+. b) 2-Chloro-4-methylpyridine-N-oxide hydrochloride 15 A mixture of 2-chloro-4-methylpyridine (10.0 g, 78.3 mmole) and 34% peracetic acid (76.05 g, 91.0 mmole) in glacial AcOH (10 mL) was heated at 70°C for 3 hr. Thereaction mixture was cooled, conc. HCl (35 mL) was added, and the mixture wasconcentrated on the rotavap. Recrystallization from n-butanol followed by trituration withether gave the title compound (7.16 g, 51%) as a white solid: MS (ES) m/e 143.9 (M + 20 H)+. c) 2-[(3-Hydroxy- l-propyl)amino]-4-methylpyridine-N-oxide A mixture of 2-chloro-4-methylpyridine-N-oxide hydrochloride (7.16 g, 39mmole), 3-aminopropanol (6.01 g, 80 mmole), and NaHCÛ3 (16.8 g, 200 mmole) in tert- 25 amyl alcohol (50 mL) was heated at reflux for 19 hr. The reaction mixture was diluted withCH2CI2 (200 mL) and filtered, and the filtrate was concentrated on the rotavap.Recrystallization from C^C^Æ^O gave the title compound (5.41 g, 75%) as a yellowsolid: TLC (15% MeOH/CH2Cl2) Rf 0.44; !H NMR (400, CDCI3) δ 7.92 (d, J = 6.7, 1 H),7.28 (br t, 1 H), 6.43 (s, 1 H), 6.33 (dd, J = 6.6, 2.1 Hz, 1 H), 3.73 (t, J=5.7 Hz, 2 H), 3.47 30 (q, H=6.3 Hz, 2 H), 2.29 (s, 3 H), 1.82 - 1.88 (m, 2 H); MS (ES) m/e 183 (M+ H)+. - 35 - 011341
Dran^rntion Aa » Uuvii ~t
Préparation of 6-(methylamino)-2-pyridvlethanoI 5 a) 2-(ieri-Butoxycarbonylamino)-6-picoline A solution of 2-amino-6-picoline (21.63 g, 200 mmole) and di-rert-butyl dicarbonate (52.38 g, 240 mmole) in CH2CI2 (200 mL) was concentrated on the rotavap at50°C, and the resulting residue was allowed to rotate on the rotavap at 50°C under vacuum.After 21.5 hr, the reaction was diluted with hexanes (400 mL) and filtered through silica gel 10 (hexanes followed by 20% EtOAc/hexanes). Concentration left the title compound (41.84g, quantitative) as a light yellow oil which gradually solidified on standing: *H NMR (250MHz, CDCI3) δ 7.71 (d, J = 8.3 Hz, 1 H), 7.40 - 7.65 (m, 2 H), 6.80 (d, J = 7.5 Hz, 1 H),2.43 (s, 3 H), 1.50 (s, 9 H); MS (ES) m/e 153 (M + H - C4H8)+. 15 b) 2-[(ferf-Butoxycarbonyl)methylamino]-6-picoline
NaH (60% in minerai oil, 3.60 g, 90 mmole) was added in portions over several min to a solution of 2-(rerr-butoxycarbonylamino)-6-picoline (15.62 g, 75 mmole) andiodomethane (9.3 mL, 150 mmole) in anhydrous DMSO (75 mL) at 15°C (cool water bath).The internai température rose to 35°C. When gas évolution had subsided, the cool water 20 bath was removed and the reaction was allowed to stir at RT. After 0.5 hr, the dark yellowmixture was poured onto iceÆ^O (300 mL) and extracted with Et2Û (3 x 300 mL). Thecombined organic layers were washed sequentially with H2O (2 x 75 mL) and brine (75mL). Drying (MgSO4) and concentration left a yellow oil which was chromatographed onsilica gel (7% EtOAc/hexanes). The title compound (13.01 g, 78%) was obtained as a 25 faintly yellow oil: ’H NMR (250 MHz, CDCI3) δ 7.51 (app t, 1 H), 7.37 (d, J = 8.2 Hz, 1H), 6.86 (d, J = 7.2 Hz, 1 H), 3.38 (s, 3 H), 2.49 (s, 3 H), 1.50 (s, 9 H); MS (ES) m/e 223 (M+ H)+. c) Ethyl-6-[(rerr-butoxycarbonyl)methylamino]-2-pyridylacetate 30 LDA was prepared at 0°C under argon from diisopropylamine (19.5 mL, 139.14 mmole) and 2.5 M n-BuLi in hexanes (46.4 mL, 115.95 mmole) in dry THF (350 mL).
This solution was cooled to -78°C and a solution of 2-[(im-butoxycarbonyl)methylamino]-
6-picoline (10.31 g, 46.38 mmole) in dry THF (46 mL) was added dropwise over 10 min.Additional dry THF (2 mL) was used in transfer. The orange solution was stirred at -78°C 35 for 15 min, then diethyl carbonate (6.2 mL, 51.02 mmole) was added rapidly. The redsolution was stirred at -78°C for 15 min, then was quenched with half-saturated NH4C1(175 mL). The mixture was warmed to +5°C and extracted with EtOAc (175 mL) then with - 36- 011341 CH2CI2 (2 x 100 mL). The combined organics were washed with brine (100 mL), dried(MgSO4), and concentrated. The cloudy yellow oil was chromatographed on silica gel(15% EtOAc/hexanes) to afford the title compound (10.72 g, 79%) as a light yellow oil: JH NMR (250 MHz, CDCI3) δ 7.51 - 7.63 (m, 2 H), 6.91 - 7.03 (m, 1 H), 4.19 (q, J = 7.1 5 Hz, 2 H), 3.77 (s, 2 H), 3.38 (s, 3 H), 1.27 (t, J = 7.1 Hz, 3 H), 1.51 (s, 9 H); MS (ES) m/e295 (M + H)+. d) Ethyl-6-(methylamino)-2-pyridyIacetate A solution of ethyl-6-[(rert-butoxycarbonyl)methylamino]-2-pyridylacetate (10.72 10 g, 36.42 mmole) in anhydrous dioxane (91 mL) was cooled to the point of partial crystallization of the solvent, and 4 M HCl/dioxane (91 mL, 364.2 mmole) was added. Thesolution was warmed to RT and stirred for 17 hr, then was concentrated. The resulting lightyellow solid was slurried with C^C^/toluene and reconcentrated to leave the titlecompound (8.48 g, quantitative) as a light yellow powder. ^H NMR (250 MHz, CD3OD) δ
15 7.84 (dd, J = 9.0, 7.2 Hz, 1 H), 6.96 (d, J = 9.0 Hz, 1 H), 6.78 (d, J = 7.2 Hz, ï H), 4.22 (q, J = 7.1 Hz, 2 H), 3.93 (s, 2 H), 3.05 (s, 3 H), 1.27 (t, J = 7.1 Hz, 3 H); MS (ES) m/e 195 (M +H)+. e) 6-(Methylamino)-2-pyridylethanol 20 A solution of 1.0 M L1AIH4 in THF (95 mL, 95 mmole) was added dropwise to a mechanically stirred suspension of ethyl-2-(methylamino)-6-pyridylacetate (7.34 g, 31.82mmole) in dry THF (64 mL) at 0°C under argon. The addition was done slowly until gasévolution subsided, then the remaining solution was added rapidly. Addition required 5 - 7min. The reaction was warmed to RT and stirred for 45 min, then was heated to reflux. 25 After 10 min, the reaction was cooled to 0°C and worked up by sequential dropwise addition of H2O (3.6 mL), 15% NaOH (3.6 mL), and H2O (10.8 mL). The mixture wasstirred for 15 min at 0°C and 15 min at RT, then was filtered through a Buchner funnel.
The filter pad was washed with plenty of THF, and the filtrate was concentrated. Theresidue was reconcentrated from toluene, then was chromatographed on silica gel (5% 30 MeOH in 1:1 EtOAc/CHC^) to afford the title compound (3.23 g, 67%) as a yellow oilwhich solidified to a waxy solid: ^H NMR (250 MHz, CDCI3) δ 7.36 (dd, J = 8.3, 7.3 Hz, 1 H), 6.42 (d, J = 7.3 Hz, 1 H), 6.26 (d, J = 8.3 Hz, 1 H), 4.93 - 5.28 (m, 1 H), 4.38 - 4.60(m, 1 H)r, 3.96 (t, J = 5.4 Hz, 2 H), 2.90 (d, J = 5.2 Hz, 3 H), 2.84 (t, J = 5.4 Hz, 2 H); MS(ES) m/e 153 (M + H)+. 35 -37 - 011341
Qrxar\qrof-îz\r» * ivpuiuuvn -j
Préparation of 2-(ethylamino)-4-thiazoleethanol 5 a) Ethyl 2-acetylamino-4-thiazoIeacetate
Ethyl 2-amino-4-thiazoleacetate (3.72 g, 20 mmole) was taken up in acetic acid (4 mL) and acetic anhydride (4 mL), and the resulting suspension was heated at reflux for 3hr. Concentration and flash chromatography on silica gel (5% MeOH/CH2H2) gave thetitle compound (4.1 g, 91%) as a white solid: MS (ES) m/e 229 (M + H)+. 10 b) 2-(EthyIamino)-4-thiazoleethanol
To a stirred solution of 1.0 M LiAIH4 in THF (179 mL, 179 mmole) was addeddropwise a solution of ethyl 2-acetylamino-4-thiazoleacetate (4.4 g .17.9 mmole) in THF(50 mL). After complété addition, the reaction mixture was heated at reflux for 3 hr, then 15 was worked up by sequential addition of H2O (0.7 mL), 10% NaOH (0.7 mL), and H2O(2.1 mL). The resulting mixture was filtered through celite® and the filtrate wasconcentrated. Purification by flash chromatography on silica gel (5% MeOH/CH2H2) gavethe title compound (1.6 g, 53%) as an amber oil: MS (ES) m/e 173 (M + H)+. 20 Préparation 6
Préparation of 6-amino-2-pyridylethanol a) 6-Amino-2-pyridylethanol 25 A solution of 2-[2-(4-methoxybenzylamino)pyridin-6-yl]ethanol (0.95 g, 3.7 mmole), prepared according to the procedure of WO 95/32710, in 6 N HCl was heated at60°C. After 16 hr, the reaction was concentrated in vacuum and the residue was madebasic with dry KOH. The resulting mixture was extracted with MeOH, and the MeOHextracts were dried (MgSO4) and concentrated. Flash chromatography on silica gel (5% 30 MeOH/CH2H2) gave the title compound (0.2 g, 40%) as a pale yellow oil: MS (ES) m/e139(M + H)+. -38- 011341
Préparation 7
Préparation of 3-(4-nitrobenzyloxycarbonylamino)-l-propanol 5 a) 3-(4-Nitrobenzyloxycarbonyl)amino-l-propanol
To a suspension stirred under argon at room température of 4-nitrobenzyl chloroformate (5 g, 23 mmol) and triethylamine (6.4 mL, 46 mmol) in THF (25 mL) wasadded 3-amino-l-propanol (1.9 mL, 26 mmol). The resulting mixture was stirred for 72 hr,then was concentrated. The residue was purified by chromatography on silica gel (0.5 - 2% 10 MeOH/CH2Cl2) to give the title compound (2 g, 34%) as a pale yellow oil: MS (ES) 255.3(M+H)+.
Préparation 8 15 Préparation of l-f(3-hydroxv-l-propvl)amino1isoquinoline-N-oxide a) 1-Chloroisoquinoline N-oxide 1-Aminoisoquinoline N-oxide hydrochloride (Deady, L. W. SyntheticCommunications 1977, 509-514) was converted to 1-chloroisoquinoline N-oxide using 20 potassium nitrite and conc. HCl according to the general method described in the literature(Brown, E. V. J. Amer. Chem. Soc. 1957, 79, 3565-3566). The title compound wasprepared as a light brown solid: MS (ES) m/e 179.9 (M + H)+. b) l-[(3-Hydroxy-l-propyl)amino]-isoquinoline N-oxide 25 According to the procedure of Préparation 1 (a), except substituting 1- chloroisoquinoline N-oxide for the 2-chloropyridine-N-oxide hydrochloride, the titlecompound was prepared as an amber solid: MS (ES) m/e 219.1 (M + H)+.
Préparation 9 30
Préparation of 2-fN-(3-methanesulfonyloxv-l-propvl)-N-(ferf- butoxycarbonyl)amino1pvridine-N-oxide a) 2-[N-(3-Hydroxy-l-propyl)-N-(zerr-butoxycarbonyl)amino]pyridine-N-oxide 35 A solution of 2-[(3-hydroxy-l-propyl)amino]pyridine-N-oxide (8.0 g, 47.6 mmol) in rerr-BuOH (80 mL) was treated with di-ferr-butyl dicarbonate (11.4 g, 55.3 mmol).
After 18h, the solution was concentrated and the residue was triturated with hexane. The - 39- rçsuking soüd was dried in vacuo to give the title compound (12.5 g, 98%) - solid: MS (ES) m/e 269.3 (M + H)+. 011341 b) 2-[N-(3-Methanesulfonyloxy- l-propyl)-N-(ferr-butoxycarbonyl)amino]pyridine-N-5 oxide
Methanesulfonyl chloride (0.17 mL, 2.20 mmole) was added dropwise to a solutionof 2-[N-(3-hydroxy-l-propyl)-N-(feri-butoxycarbonyl)amino]pyridine-N-oxide (0.50 g, 1.86 mmole) and pyridine (0.23 mL, 2.84 mmole) in CHCI3 (5 mL, dried over K2CO3) at
_ X 0°C. When complété by TLC, the reaction was diluted with CHCI3, washed with ice water,10 dried (Na2SC>4), and concentrated. Silica gel chromatography (10% MeOH/CHC^) gave the title compound (0.41 g, 64%) as a colorless oil: NMR (250 MHz, CDCI3) δ 8.25 (dd, J = 6.0,1.9 Hz, 1 H), 7.25 (m, 4 H), 4.35 (t, J = 6.2 Hz, 2 H), 3.75 (t, J = 6.6 Hz, 2 H),3.00 (s, 3 H), 2.00 (m, 2 H), 1.40 (s, 9 H). Unchanged 2-[N-(3-hydroxy-l-propyl)-N-(rerr-butoxycarbonyl)amino]pyridine-N-oxide (0.18 g, 36%) could also be recovered from the 15 chromatographie purification.
Préparation 10
Préparation of ethyl (±)-10,l l-dihvdro-3-hvdroxy-5H-dibenzoia,d1cycloheptene-10-acetate 20 a) 6-Methoxy-l-phenyIindene A solution of 3.0 M phenylmagnesium bromide in Et2O (680 mL, 2.04 mole) underargon at ambient température was diluted with Et2Û (700 mL) with stirring, and a solutionof 6-methoxy-l-indanone (277 g, 1.71 mole) in THF (1400 mL) was added dropwise over 1 25 hr. The reaction mixture was stirred for 2 h at ambient température and then was pouredwith stirring into saturated NH4CI (2.8 L). H2O (1.4 L) was added, and the organic phaseseparated. The aqueous phase was extracted with Et2Û (2 x l L), and the combined organicextracts were concentrated to give crude 6-methoxy-l-phënyl-l-indanol (445 g) as a brownoil. This oil was dissolved in toluene (2.5 L), and p-toluenesulfonic acid monohydrate 30 (12.3 g, 0.065 mole) was added. The solution was stirred and heated at reflux for 16 hr using a Dean-Stark trap with a condenser. H2O collection was minimal after 2 h andtotaled 28 mL. The solution was cooled and extracted sequentially with 5 % aqueousNa2CC>3 (1 L) and H2O (2x1 L). The organic layer was concentrated to give a darkbrown oil (400 g). This oil was distilled under vacuum to give the title compound (298.2 g, 35 79%) as a yellow oil: bp 152 - 190°C/2.0 Torr; TLC (10% EtOAc/hexanes) Rf 0.75. -40- 011341 b) 2-Benzoyl-4-methoxyphenylacetic acid
Acetone (4.2 L) was chilled to 10°C, and a solution of 6-methoxy-l-phenylindene(271 g, 1.22 mole) in acetone (1.8 L) was added over 1.5 hr concurrently with Jones reagent( 1.8 L, prepared front C1O3 (470 g, 4.70 mole), H2O (1 L), and conc H2SO4 (405 mL)). 4 5 % Aqueous OSO4 (153 mL) was added to the resulting mixture in two portions, one at the
onset of addition and the second at the mid-point of the addition, maintaining thetempérature of the reaction mixture below 15°C. Following the addition, the reactionmixture was warmed to 22°C and stirred for 1.5 h, during which time a mild exothermincreased the température to 28°C. The reaction mixture was then cooled to below 20°C 10 and isopropanol (1 L) was added, dropwise initially and rapidly after the initial exothermdiminished. Stirring became difficult during this phase. The température reached 32°Cduring the isopropanol addition. H2O (2 L) was added and the mixture was transferred to aseparatory funnel. Additional H2O was added to dissolve the precipitated chromous acid,and the mixture was extracted with CH2CI2 (2 L). The organic (upper) layer was separated 15 and the aqueous phase was extracted with CH2CI2 (2 x 1 L). The combined CH2CI2 extracts were washed sequentially with H2O (2 L) and saturated brine (2 L), and then wereconcentrated to give a moist gray solid (416 g). This was triturated with a mixture ofacetone and EtOAc and filtered and dried to give the title compound (225.4 g, 71%) as anoff-white solid: mp 158-159°C. 20 c) 2-Benzyl-4-methoxyphenylacetic acid 2-Benzoyl-4-methoxyphenylacetic acid (215.5 g, 0.80 mole) was divided into twoequal portions, and each was dissolved in glacial AcOH (1.5 L) in a 2.5 L pressure bottle. 5% Pd/C (10 g, 0.0048 mole) was added to each, and each mixture was shaken at ambient 25 température under hydrogen on a Parr apparatus. After 2.5 hr, the mixtures were filtered toremove the catalyst, and the filter pads were washed with EtOAc. The combined filtrâteswere concentrated to give the title compound (215 g, quantitative) as a heavy yellow oilwhich crystallized on standing: *H NMR (250 MHz, CDCI3) δ 7.05 - 7.35 (m, 6 H), 6.77(dd, J = 8.3, 2.7 Hz, 1 H), 6.71 (d, J = 2.7 Hz, 1 H), 4.00 (s, 2 H), 3.76 (s, 3 H), 3.54 (s, 2 30 H). d) 10,1 l-Dihydro-3-methoxy-5H-dibenzo[a,d]cyclohepten-10-one A solution of 2-benzyl-4-methoxyphenylacetic acid (215 g of crude material thatcontained 204.6 g (0.80 mole) of pure material) in CH2CI2 (1 L) was stirred under argon at 35 ambient température, and DMF (1 mL) was added, followed by oxalyl chloride (400 mL,4.59 mole). The oxalyl chloride was added over 1 hr, dropwise initially to control thevigorous gas évolution. The solution was stirred for 16 h at ambient température and then -41 - 011341 ΙΟ was concentrated to give the crade acid chloride (207.7 g, 0.756 mol, 95 %) as a yeîlowliquid. This liquid was dissolved in CH2CI2 to a total volume of 500 mL, and the solutionand AICI3 (100.8 g, 0.756 mol) were added concurrently over 1 hr to CH2CI2 (3.7 L) withstirring under argon at ambient température. The température was 28°C at the completionof the addition. The reaction mixture was stirred for 16 h at ambient température, duringwhich time a solid precipitated. H2O (1 L) was added, initially dropwise, over a period of30 min. The mixture was then separated and the organic phase was washed sequentiallywith H2O (1 L) and 5 % aqueous NaHCC>3 (1 L). The CH2CI2 solution was then s. concentrated to give a yellow solid (175.3 g). Recrystallization from EtOAc/hexane gavethe title compound (128 g, 71%): mp 107 - 109°C. e) Ethyl (±)-10,l l-dihydro-10-hydroxy-3-methoxy-5H-dibenzo[a,d]cycloheptene-10-acetate A 1.0 M solution of lithium bis(trimethylsilyl)amide in hexanes (1282 mL, 1.282 15 mole) was added to THF (4.0 L) at -70°C under argon, then EtOAc (146 mL, 1.49 mole)was added dropwise over 20 min. The reaction mixture was allowed to stir for 15 min, thenΝ,Ν,Ν’,Ν’-tetramethylethIylenediamine (378 mL, 2.5 mole) was added over 20 min. Thereaction mixture was stirred for 10 min, then a solution of 10,1 l-dihydro-3-methoxy-5H-dibenzo[a,d]cyclohepten-10-one (119.2 g, 0.50 mol) in anhydrous THF (1.26 L) was added 20 dropwise over 40 min. The température was maintained below -65°C during ail of theseadditions. The reaction mixture was stirred for 20 min at -65 to -70°C and then was pouredinto saturated aqueous NH4CI (6.2 L) with vigorous stirring. The organic layer wasseparated and the aqueous phase was extracted with EtOAc (2 x 1 L). The combinedorganic extracts were washed with H2O (2 x 1 L) and then were concentrated to give a light 25 brown oil (175 g). Thin-layer chromatography (20 % EtOAc/hexanes) showed Rf 0.5major (desired product) and Rf 0.7 minor (recovered ketone). The crade product waschromatographed on silica gel (2 kg, 10 % EtOAc/hexanes) to afford the title compound(101 g, 61 %) as a yellow oil: ^H NMR (250 MHz, CDCI3) δ 7.63 (d, J = 7.7 Hz, 1 H), 7.00 - 7.30 (m, 4 H), 6.80 (d, J = 2.6 Hz, 1 H), 6.69 (dd, J = 8.2, 2.6 Hz, 1 H), 3.95 - 4.35 30 (m, 2 H), 4.07 (s, 2 H), 3.76 (s, 3 H), 3.68 (s, 1 H), 3.64 (d, J = 14.2 Hz, 1 H), 3.35 (d, J = 14.2 Hz, 1 H), 2.79 (d, J = 16.0 Hz, 1 H), 2.66 (d, J = 16.0 Hz, 1 H), 1.22 (t, J = 7.2 Hz, 3H). f) Ethyl (±)-10,l l-dihydro-3-methoxy-5H-dibenzo[a,d]cycloheptene-10-acetate 35 Ethyl (±)-10,l l-dihydro-10-hydroxy-3-methoxy-5H-dibenzo[a,d]cycloheptene-10- acetate (101 g, 0.31 mole) was dissolved in glacial acetic acid (1.8 L) and 12 N HCl (28.5mL, 0.34 mole) was added. The mixture was placed in a 2.5 L pressure bottle containing -42- 011341 5% Pd/C (20 g, 0.0094 mole), and the resulting mixture was shaken at 35°C underhydrogen on a Parr hydrogénation apparatus equipped with a jacket heater. After 18 hr, thereaction was cooled to ambient température, and the catalyst was removed by filtration.
The filtrate was concentrated to give a light yellow oil (85.1 g). This was chromatographed 5 on silica gel (2 kg, step-gradient with 5 % to 10 % EtOAc/hexanes) to afford the titlecompound (69.1 g, 72%) as an oil: ÏH NMR (250 MHz, CDCI3) δ 7.05 - 7.22 (m, 4 H), 7.01 (d, J = 8.2 Hz, 1 H), 6.76 (d, J = 2.7 Hz, 1 H), 6.67 (dd, J = 8.2, 2.7 Hz, 1 H), 4.30 (d, J= 15.0 Hz, 1 H), 4.11 - 4.25 (m, 2 H), 3.85 (d, J = 15.0 Hz, 1 H), 3.70 - 3.90 (m, 1 H), 3.77(s, 3 H), 3.31 (dd, J = 15.0,4.1 Hz, 1 H), 2.93 (dd, J = 15.0,9.2 Hz, 1 H), 2.64 (dd, J = 15.6, 10 5.0 Hz, 1 H), 2.52 (dd, J = 15.6, 9.3 Hz, 1 H), 1.27 (t, J = 7.1 Hz, 3 H). g) Ethyl (+)-10,l l-dihydro-3-hydroxy-5H-dibenzo[a,d]cycloheptene- 10-acetate A solution of ethyl (±)-10,l l-dihydro-3-methoxy-5H-dibenzo[a,d]cycloheptene-10- acetate (8.5 g, 0.027 mole) in CH2CI2 (150 mL) was chilled to -10°C with stirring under 15 argon. Ethanethiol (10.7 mL, 0.144 mole) was added, followed by AICI3 (20.6 g, 0.154mole) in two portions over 15 min. An exotherm increased the température to 0°Cfollowing the additions, and the température was then increased to 25°C using a water bath.The réaction mixture was stirred at 25 to 30°C for 2.25 hr, at which point it was poured intoice-H2O. The organic layer was separated, methanol (100 mL) was added, and the mixture 20 was extracted with CH2CI2 (2 x 50 mL). The combined CH2CI2 extracts were washedwith H2O (250 mL) and then were concentrated to give a viscous oil (8.6 g). This wastaken up in Et2Û (150 mL) and the ether was boiled off while replacing it with hexane.
The desired phénol first separated as an oil which crystallized on stirring at ambienttempérature. Two crops of solid were collected to afford the title compound (7.1 g, 89 %): 25 mp 110 - 112°C.
Préparation 11 HPLC séparation of the enantiomers of ethyl (i)-10.1 l-dihvdro-3-hydroxv-5H- 30 dibenzoia,dlcycloheptene-10-acetate a) Ethyl (R)-(+)-10,1 l-dihydro-3-hydroxy-5H-dibenzo[a,d]cycloheptene- 10-acetate andethyl (S)-(-)-10,1 l-dihydro-3-hydroxy-5H-dibenzo[a,d]cycloheptene-10-acetate
Ethyl (±)-10,l l-dihydro-3-hydroxy-5H-dibenzo[a,d]cycloheptene-10-acetate was 35 resolved into its enantiomers using the following conditions: Daicel Chiralcel OJ® column(21.2 x 250 mm), 20% éthanol in hexane mobile phase, 15 mL/min flow rate, uv détectionat 254 nm, 140 mg injection; Ir for ethyl (S)-(-)-10,1 l-dihydro-3-hydroxy-5H- -43- 011341 dibenzo[a,d]cycîohexer.e-ÎO-acetate — 10.4 min.; îr forethyl (R)-(+)-iO,l î-dihydro-3-hydroxy-5H-dibenzo[a,d]cyclohexene-10-acetate = 13.1 min.
Préparation 12 5
Préparation of ethyl (±)-10,l l-dihydro-7-fluoro-3-hydroxy-5H-dibenzora,d1cycloheptene-10-acetate a) 1 -(3-Fluoropheny l)-6-methoxy-1 -indanol 10 According to the procedure of Préparation 10 (a), except substituting 3- fluorophenylmagnesium bromide for the phenylmagnesium bromide, the title compoundwas obtained as an amber oil: MS (ES) m/e 276.0 (M + H)+. b) 1 -(3-Fluoropheny l)-6-methoxy indene 15 According to the procedure of Préparation 10 (a), except substituting l-(3- fluorophenyl)-6-methoxy-l-indanol for the 6-methoxy-l-phenyl-l-indanol, the titlecompound was obtained as a colorless oil following silica gel chromatography (4%EtOAc/hexanes): MS (ES) m/e 241.1 (M + H)+. 20 c) 2-(3-Fluorobenzoyl)-4-methoxyphenylacetic acid
According to the procedure of Préparation 10 (b), except substituting 2-(3- fluorophenyl)-6-methoxyindene for the 6-methoxy-l-phenylindene, the title compound wasobtained as a white solid: MS (ES) m/e 289.2 (M + H)+. 25 d) 2-(3-Fluorobenzyl)-4-methoxyphenylacetic acid
According to the procedure of Préparation 10 (c), except substituting 2-(3- fluorobenzoyl)-4-methoxyphenylacetic acid for the 2-benzoyl-4-methoxyphenylacetic acid,the title compound was obtained as a colorless oil: MS (ES-) m/e 273.2 (M - H)'. 30 e) 10,1 l-Dihydro-7-fluoro-3-methoxy-5H-dibenzo[a,d]cyclohepten-10-one
According to the procedure of Préparation 10 (d), except substituting 2-(3- fluorobenzyl)-4-methoxyphenylacetic acid for the 2-benzyl-4-methoxyphenylacetic acid,the title compound was obtained as a white solid: Mp 129 - 130°C; MS (ES) m/e 279.2 (M+ Na)+. 35 011341 -44- f) Ethyl (±)-10,l l-dihydro-7-fluoro-10-hydroxy-3-methoxy-5H-dibenzo[a,d]cycloheptene-10-acetate
According to the procedure of Préparation 10 (e), except substituting 10,11-dihydro-7-fluoro-3-methoxy-5H-dibenzo[a,d]cyclohepten-10-one for the 10,1 l-dihydro-3- 5 methoxy-5H-dibenzo[a,d]cyclohepten-10-one, the title compound was obtained followingsilica gel chromatography (8% EtOAc/hexanes): MS (ES) m/e 362.2 (M + NH4)+. g) Ethyl (±)-10,1 l-dihydro-7-fluoro-3-methoxy-5H-dibenzo[a,d]cycloheptene- 10-acetate
According to the procedure of Préparation 10 (f), except substituting ethyl (±)- 10 10,11 -dihydro-7-fluoro-10-hydroxy-3-methoxy-5H-dibenzo[a,d]cycloheptene-10-acetate for the ethyl (±)-10,l l-dihydro-10-hydroxy-3-methoxy-5H-dibenzo[a,d]cycloheptene-10-acetate, the title compound was obtained as a colorless oil following silica gelchromatography (10% EtOAc/hexanes): MS (ES) m/e 329.2 (M + H)+. 15 h) Ethyl (±)-10,l l-dihydro-7-fluoro-3-hydroxy-5H-dibenzo[a,d]cycloheptene-10-acetateAccording to the procedure of Préparation 10 (g), except substituting ethyl (±)- 10,1 l-dihydro-7-fluoro-3-methoxy-5H-dibenzo[a,d]cycloheptene-10-acetate for the ethyl(±)-10,ll-dihydro-3-methoxy-5H-dibenzo[a,d]cycloheptene-10-acetate, the title compoundwas obtained as a white solid following silica gel chromatography (1% MeOH/CH2Cl2): 20 MS (ES) m/e 315.0 (M + H)+, 332.0 (M + NH4)+.
Préparation 13
Préparation of ethyl (±)-10,l l-dihydro-2-(dimethylamino)methvl-7-fluoro-3-hydroxv-5H- 25 dibenzoi a.dlcycloheptene-10-acetate a) Ethyl (±)-10,1 l-dihydro-2-(dimethylamino)methyl-7-fluoro-3-hydroxy-5H-dibenzo[a,d]cycloheptene-10-acetate
To a solution of ethyl (±)-10,11 -dihydro-7-fluoro-3-hydroxy-5H-dibenzo
30 [a,d]cycloheptene-10-acetate (0.4 g, 1.33 mmol) in 95% éthanol containing 2 M dimethylamine in MeOH (1.0 mL) was added 37% aqueous formaldéhyde solution (0.5mL) at RT under argon. After 20 hr, the reaction was heated to reflux for 5 hr, then wasconcentrated on the rotavap. The residue was partitioned between H2O and Et2Û, and thelayers were separated. The aqueous layer was extracted with Et2Û, and the combined 35 organic layers were washed with brine, dried (MgSO4), and concentrated on the rotavap togive the title compound (330 mg, 67%) as a colorless oil: NMR (400 MHz, CDCI3) δ 7.20 (m, 1 H), 6.88 (m, 2 H), 6.67 (s, 2 H),4.25 (d, J = 15.1 Hz, 1 H), 4.18 (q, 2 H), 3.78 -45- 011341 (m, 1 H), 3.74 (d, J — 15. ! Hz, 1 H), 3.55 (s, 2 H), 3.20 (dd, ï H), 2.80 (dd, 1 H), 2.60 (dd, 1H), 2.53 (dd, 1 H), 2.29 (S, 6 H), 1.27 (t, 3 H); MS(ES) m/e 372.3 (M + H)+.
Préparation 14 5
Préparation of ethvl (±)-10,l l-dihvdro-3-hvdroxy-2-methyl-5H-dibenzo[a,dlcycloheptene- 10-acetate a) Ethyl (±)-10,l l-dihydro-2-formyl-3-methoxy-5H-dibenzo[a,d]cycIoheptene-10-acetate 10 POCI3 ( 17 mL) was added dropwise to a solution of ethyl (±)-10,11 -dihydro-3- methoxy-5H-dibenzo[a,d]cycloheptene-10-acetate (1.0 g, 3 mmol) in dry DMF (40 mL) atRT under argon, and the dark solution was heated at 90°C for 48 hr. The reaction wasconcentrated on the rotavap and the residue was partitioned between H2O and EtOAc. Theorganic layer was separated, dried (MgSO4), and concentrated on the rotavap. The residue 15 was reconcentrated from xylenes (to remove any remaining DMF) then was chromatographed on silica gel (7% EtOAc in hexanes) to afford the title compound (230mg, 21%) as a colorless oil: MS (ES) m/e 339.3 (M + H)+. b) Ethyl (±)-10,11 -dihydro-3-methoxy-2-methyl-5H-dibenzo[a,d]cycloheptene-10-acetate 20 A mixture of ethyl (±)-10,11 -dihydro-2-formyl-3-methoxy-5H- dibenzo[a,d]cycloheptene-10-acetate (220 mg, 0.65 mmol), 10% Pd/C (90 mg), glacialHOAc (15 mL), and conc HCl (2 mL) was shaken at RT under hydrogen (60 psi). After 20hr, the mixture was filtered through celite®, and the filtrate was concentrated to afford thetitle compound (200 mg, 95%) as a colorless oil: MS (ES) m/e 325.2 (M + H)+. 25 c) Ethyl (±)-10,1 1-dihydro-3-hydroxy-2-methyl-5H-dibenzof a,d]cycloheptene- 10-acetate
To dry CH2CI2 (30 mL) cooled in an ice bath was added diethylsulfide (0.38 mL,3.3 mmol) followed by AICI3 (438 mg, 3.3 mmol). To this solution was added dropwise asolution of ethyl (±)-10,l l-dihydro-3-methoxy-2-methyl-5H-dibenzo[a,d]cycloheptene-10- 30 acetate (200 mg, 0.6 mmole) in dry CH2CI2 (6 mL), and the resulting mixture was stirredat RT for 2 hr. The reaction was quenched with 1.0 N HCl (10 mL), and the layers wereseparated. The organic layer was dried (MgSC>4) and concentrated on the rotavap to givethe title compound (100 mg, 56%) as a colorless oil: MS (ES) m/e 311.2 (M + H)+. -46- 011341
Préparation 15
Préparation of ethyl (+)-10,1 l-dihvdro-3-hydroxy-6-methyl-5H-dibenzofa,dkvcloheptene- 10-acetate 5 a) 6-Methoxy-1 -(2-methylphenyl)-1 -indanol
According to the procedure of Préparation 10 (a), except substituting 2-methylphenylmagnesium bromide for the phenylmagnesium bromide, the title compoundwas obtained as an oil: MS (ES) m/e 277.0 (M + Na)+. 10 b) 6-Methoxy- l-(2-methylphenyl)indene
According to the procedure of Préparation 10 (a), except substituting 6-methoxy-l-(2-methylphenyl)-1 -indanol for the 6-methoxy-l-phenyl-l-indanol, the title compound wasobtained as a colorless oil following silica gel chromatography (3% EtOAc/hexanes): MS 15 (ES) m/e 237.2 (M + H)+. c) 4-Methoxy-2-(2-methylbenzoyl)phenylacetic acid
According to the procedure of Préparation 10 (b), except substituting 6-methoxy-l-(2-methylphenyl)indene for the 6-methoxy-l-phenylindene, the title compound was 20 obtained as a viscous oil: MS (ES) m/e 285.3 (M + NH4)+. d) 4-Methoxy-2-(2-methylbenzyl)phenylacetic acid
According to the procedure of Préparation 10 (c), except substituting 4-methoxy-2-(2-methylbenzoyl)phenylacetic acid for the 2-benzoyl-4-methoxyphenylacetic acid, the title 25 compound was obtained as a viscous oil: MS (ES) m/e 288.2 (Μ + NH4)4·. e) 10,1 l-Dihydro-3-methoxy-6-methyl-5H-dibenzo[a,d]cyclohepten- 10-one
According to the procedure of Préparation 10 (d), except substituting 4-methoxy-2-(2-methylbenzyl)phenylacetic acid for the 2-benzyl-4-methoxyphenylacetic acid, the title 30 compound was obtained as a white solid following silica gel chromatography (6%EtOAc/hexanes): MS (ES) m/e 253.0 (M + H)+. f) Ethyl (±)-10,11 -dihydro-10-hydroxy-3-methoxy-6-methy 1-5H-dibenzo[a,d]cy cloheptene-10-acetate 35 According to the procedure of Préparation 10 (e), except substituting 10,11- dihydro-3-methoxy-6-methyl-5H-dibenzo[a,d]cyclohepten- 10-one for the 10,1 l-dihydro-3- 011341 -47- methoxy-5H-dibenzo[a,d]cyc!ohepten-!0-one, the title compound was obîained foilowingsilica gel chromatography (8% EtOAc/hexanes): MS (ES) m/e 358.2 (M + NH4)+. g) Ethyl (±)-10,l l-dihydro-3-methoxy-6-methyl-5H-dibenzo[a,d]cycloheptene-l 0-acetate 5 According to the procedure of Préparation 10 (f), except substituting ethyl (±)- 10,1 l-dihydro-10-hydroxy-3-methoxy-6-methyl-5H-dibenzo[a,d]cycloheptene-10-acetatefor the ethyl (±)-10,l l-dihydro-10-hydroxy-3-methoxy-5H-dibenzo[a,d]cycloheptene-10-acetate, the title compound was obtained as a colorless oil foilowing silica gelchromatography (5% EtOAc/hexanes): MS (ES) m/e 325.3 (M + H)+. 10 h) Ethyl (±)-10,l l-dihydro-3-hydroxy-6-methyl-5H-dibenzo[a,d]cycloheptene-10-acetate
According to the procedure of Préparation 10 (g), except substituting ethyl (±)- 10,1 l-dihydro-3-methoxy-6-methyl-5H-dibenzo[a,d]cycloheptene-10-acetate for the ethyl(±)-10,l l-dihydro-3-methoxy-5H-dibenzo[a,d]cycIoheptene-10-acetate, the title compound 15 was obtained as a white solid after trituration with MeOH: MS (ES) m/e 311.2 (M + H)+.
Préparation 16
Préparation of ethyl (ί)-ΙΟ,Ι l-dihydro-3-i3-(4-nitro-l-oxopyridin-2-ylamino)-l- 20 propy loxvl-5H-dibenzoia,dlcvcloheptene-10-acetate a) Ethyl (±)-10,1 l-dihydro-3-[3-(4-nitro- l-oxopyridin-2-ylamino)- l-propyloxy]-5H-dibenzo[a,d]cycloheptene-10-acetate A solution of 2-[(3-hydroxy-l-propyl)amino]-4-nitropyridine-N-oxide (0.85 g, 4 25 mmole) and diethyl azodicarboxylate (0.63 mL, 4 mmole) in anhydrous DMF (10 mL) wasadded dropwise to a solution of ethyl (±)-!0,l l-dihydro-3-hydroxy-5H-dibenzo[a,d]cycloheptene-10-acetate (0.59 g, 2 mmole) and triphenylphosphine (1.10 g, 4.2mmole) in anhydrous DMF (10 mL) at RT under argon.; After 23 hr, the reaction wasconcentrated and the residue was reconcentrated from xylenes (2 x). Silica gel 30 chromatography (gradient: 1:1 EtOAc/hexanes, then EtOAc, then 5% MeOH in 1:1
EtOAc/CHCl3) gave crude title compound. Unchanged (±)-10,l l-dihydro-3-hydroxy-5H-dibenzo[a,d]cycloheptene-10-acetate could be recovered from the 1:1 EtOAc/hexanesfractions. Rechromatography of the crude title compound (3% MeOH in 1:1EtOAc/CHCl3) gave clean title compound (0.72 g, 73%) as a yellow foam: TLC (10% 35 MeOH in 1:1 EtOAc/CHCl3) Rf 0.59; NMR (250 MHz, CDC13) δ 8.19 (d, J = 7.1 Hz, 1H), 7.46 (d, J = 2.9 Hz, 1 H), 7.35 (dd, J = 7.1, 2.9 Hz, 1 H), 7.00 - 7.30 (m, 5 H), 7.00 (d, J= 8.2 Hz, 1 H), 6.81 (d, J = 2.6 Hz, 1 H), 6.70 (dd, J = 8.2, 2.6 Hz, 1 H), 4.29 (d, J = 15.1 011341 -48-
Hz, 1 H), 4.18 (q, J = 7.1 Hz, 2 H), 4.08 (t, J = 5.5 Hz, 2 H), 3.86 (d, J = 15.1 Hz, 1 H), 3.72- 3.90 (m, 1 H), 3.59 (q, J = 6.3 Hz, 2 H), 3.30 (dd, J = 15.0,4.2 Hz, 1 H), 2.93 (dd, J = 15.0, 9.3 Hz, 1 H), 2.64 (dd, J = 15.6, 5.1 Hz, 1 H), 2.51 (dd, J = 15.6, 9.3 Hz, 1 H), 2.10 - 2.30 (m, 2 H), 1.27 (t, J = 7.1 Hz, 3 H); MS (ES) m/e 492 (M + H)+. 5
Préparation 17
Préparation of ethvl (S)-10,l l-dihvdro-3-[3-(4-nitro-l-oxopyridin-2-ylamino)-l- propyloxy 1-5H-dibenzof a.dlc vcloheptene-10-acetate 10 a) Ethyl (S)-10,11 -dihydro-3-[3-(4-nitro-1 -oxopyridin-2-ylamino)-1 -propyloxy]-5H-dibenzo[a,d]cycloheptene-l O-acetate
According to the procedure of Préparation 16, except substituting ethyl (S)-10,11-dihydro-3-hydroxy-5H-dibenzo[a,d]cycloheptene-l 0-acetate for the ethyl (±)-10,11 - 15 dihydro-3-hydroxy-5H-dibenzo[a,d]cycloheptene-10-acetate, the title compound was prepared: MS (ES) m/e 492 (M + H)+.
Préparation 18 20 Préparation of 10.1 l-Dihydro-3-methoxv-5H-dibenzofa.dlcvclohepten-10-one a) 2-Benzyl-4-methoxyphenylacetic acid A solution of 2-benzoyl-4-methoxyphenylacetic acid (13.0 g, 0.048 mol), prepared by the method of J. Med. Chem. 1981,24,998, in glacial acetic acid (600 mL) was treated 25 under argon with 4.3 g. of 10% Pd/C and hydrogenated at 50 psi for 17 hours. The mixturewas filtered using celite® and the filtrate was concentrated and reconcentrated from tolueneand methylene chloride to give 14.2 of the title compound: NMR (400 MHz, CDCI3) δ 3.52 (s, 2H), 3.75 (s, 3H), 4.0 (s, 3H), 6.7 (m, 2H), 7.15 (m, 6H). 30 b) l0,ll-Dihydro-3-methoxy-5H-dibenzo[a,d]cyclohepten-10-one A solution of 2-benzyl-4-methoxyphenylacetic acid (14.2 g, 0.055m) in benzene (120 mL) and thionyl chloride (28 mL) was refluxed for 1 hour and concentrated. The acidchloride was dissolved in dry methylene chloride (40 mL), and the solution was addeddropwise under argon to a solution of A1C13 (14.7 g, 0.11 mol) in methylene chloride (600 35 mL). The reaction was stirred under an argon atmosphère for 2.5 hours at room température, then was quenched with ice-water (200 mL). The layers were separated, andthe organic phase was washed sequentially with 10% NaOH solution, water, and dil. HCl. -49- 011341
The resulting solution was diluted with ether (200 mL), dried over MgSO4, andconcentrated. The solid residue was triturated with ether/hexane (1:1) and 935 g of thetitle compound was collected by filtration: Mp 105 - 106°C; ’H NMR (400 MHz, CDCI3)δ 3.72 (s, 3H), 4.1 (s, 2H), 4.2 (s, 2H), 6.7 (d, 1H), 6.82 (s, 1H), 7.30 (m, 4H), 8.1 (d, 1H). 5
Préparation 19
Préparation of ethyl (±)-10,l l-dihydro-3-methoxy-5H-dibenzo[a,dlcvclohéptene-10-acetate 10 a) Ethyl (±) 3-(3-methoxyphenyl)indeneacetate
To a cold solution of 3-(3-methoxyphenyl)indene (4 g, 18 mmol), prepared by the
method of J. Med. Chem. 1981, 24, 998, in THF (15 mL) at 0°Cwas added dropwise asolution of LiN(TMS)2 (20 mL, IM in THF) over 5 min. The resulting solution was addeddropwise to a solution of ethyl bromoacetate (3.34 g, 20 mmol) in THF (15 mL) at -78°C 15 over 30 min. After 2.5 h, the mixture was quenched with saturated ammonium chloridesolution and the layers were separated. The organic layer was dried over MgSO4 andconcentrated to give the crude product which was purified by column chromatography(SiO2/2-4% EtOAc/hexane) to give title compound (1.1g): ^H NMR (400 MHz, CDCI3) 5 1.30 (t, 3H), 2.50 (m, 1H), 2.85 (m, 1H), 3.85 (s, 3H), 4.0 (m, 1H), 4.20 (q, 2H), 6.6 (s, 1H), 20 6.9 (m, 1H), 7.2 (s, 1H), 7.35 (m, 6H). b) Ethyl (±) 3-[(3-methoxybenzoyl)]phenylsuccinate A solution of ethyl (±) 3-(3-methoxyphenyl)indeneacetate (1.1 g, 3.6 mmol) inacetone (30 mL) was treated.with 4% aqueous solution of osmium tetroxide (0.5 mL) 25 followed by a dropwise addition of 1.2 M Jones reagent (5 mL, 6 mmol) according to theliterature procedure (J. Org. Chem. 1993, 58, 4745). After stirring ovemight at roomtempérature, the dark reaction mixture was quenched with isopropanol (2.5 mL), followedby sodium bisulfite (0.9 g) and water (30 mL). The prodùct was extracted with ethylacetate, washed with brine, dried over MgSC>4, and concentrated to give a solid residue. 30 Trituration with 1:1 ether / hexane gave 0.76 g of the title compound: 1H NMR (400 MHz,CDCI3) δ 1.18 (t, 3H), 2.90 (m, 1H), 3.3 (m, 1H), 3.92 (s, 3H), 4.1 (q, 2H), 4.4 (m, 1H), 4.4(d, 1H), 7.25 (m, 2H), 7.5 (m, 6H). c) Ethyl (±) 3-[(3-methoxybenzyl)]phenylsuccinate 35 A mixture of ethyl (±) 3-[(3-methoxybenzoyl)]phenylsuccinate (0.76 g., 2.1 mmol) and 10% Pd/C (0.6 g) in glacial acetic acid (35 mL) was hydrogenated at 50 psi for 17hours. The mixture was filtered using celite® and the filter pad was washed with acetic -50- 011341 acid. The filtrate was concentrated and reconcentrated from toluene and methylenechloride to give 0.65 g of the title compound: *H NMR (400 MHz, CDCI3) δ 1.20 (t, 3H), 2.20 (m, 1H), 3.0 (m, 1H), 3.74 (s, 3H), 4.1 (q, 2H), 4.18 (q, 2H),4.4 (d, 1H), 6.2 (m, 2H),7.22 (m, 6H). 5 d) Ethyl (±)-10,l l-dihydro-3-methoxy-l l-oxo-5H-dibenzo[a,d]cycloheptene-10-acetate
To a magnetically stirred solution of ethyl (±) 3-[(3-methoxybenzyl)]phenylsuccinate (0.65 g, 1.9 mmol) in dry methylene chloride (10 mL)were added DMF (0.2 mL) and oxalyl chloride (0.2 mL, 2.28 mmol). After 1.5 h, the 10 solution was added dropwise to a suspension of aluminum chloride (0.6 g, 4.5 mmol) in drymethylene chloride (15 mL). The mixture was quenched after 2 h with ice water, the layerswere separated, and the aqueous layer was extracted with methylene chloride. Thecombined organic layers were dried over MgSC>4 and concentrated. The residue waspurifïed by column chromatography (SiO2/2-4% EtOAc/hexane) to give title compound 15 (0.3 g): !h NMR (400 MHz, CDCI3) δ 1.28 (t, 3H), 2.88 (m, 1H), 3.55 (m, 1H), 3.84 (s, 3H), 3.88 (d, 1H), 4.18 (q, 2H), 4.85 (d, 1H), 4.95 (m, 1H), 5.8 (m, 2H), 7.22 (m, 4H), 8.1(s, 1H). e) Ethyl (±)-10,l l-dihydro-3-methoxy-5H-dibenzo[a,d]cycloheptene-10-acetate 20 A mixture of ethyl (±)-10,11 -dihydro-3-methoxy-11 -oxo-5H-dibenzo[a,d]cycloheptene-10-acetate (0.3 g., 0.93 mmol) and 10% Pd/C (0.3 g) in glacial acetic acid (25 mL) washydrogenated at 50 psi for 18 hours. The mixture was filtered using celite® and washedwith acetic acid. The filtrate was concentrated and reconcentrated from toluene andmethylene chloride to give 0.25 g of the title compound: NMR (400 MHz, CDCI3) δ 25 1.28 (t, 3H), 2.60 (m, 2H), 2.90 (m, 1H), 3.30 (m, 1H), 3.80 (s, 3H), 3.85 (d, 1H), 4.18 (q, 2H), 4.30 (d, 1H), 6.70 (m, 2H), 7.0 (d, 1H), 7.22 (m, 4H).
Préparation 20 30 Préparation of ethyl (i)-10,l l-dihvdro-3-hydroxy-dibenzofb.f1oxepine-10-acetate a) 4-Methoxy-2-phenoxyacetophenone
According to the procedure of Harris, T. W. et al. (7. Med. Chem. 1982, 25(7), 855 - 858), 2-fluoro-4-methoxyacetophenone (1.00 g, 5.95 mmole) was reacted with phénol to 35 give the title compound ( 1.27 g) as an oil: lH NMR (300 MHz, CDCI3) δ 7.90 (d, J = 8.8Hz, 1 H), 7.35 (m, 2 H), 7.20 (m, 1 H), 7.05 (m, 2 H), 6.70 (dd, J = 2.4, 8.8 Hz, 1 H), 6.35(d, J = 2.4 Hz, 1 H), 3.75 (s, 3 H), 2.61 (s, 3 H). -51 - 01 1341 b) 2-(4-Methoxy-2-phenoxyphenyl)-1 -morpholin-4-ylethan-1 -thione
According to the procedure of Harris, T. W., et al. (7. Med. Chem. 1982, 25(7), 855- 858), 4-methoxy-2-phenoxyacetophenone (1.69 g, 6.98 mmole), sulfur (0.36 g, 11.2 5 mmole), and morpholine (0.98 mL, 11.2 mmole) were reacted to give the title compound(1.24 g) as a white solid: MS (ES) m/e 344.0 (M + H)+. c) 2-(4-Methoxy-2-phenoxyphenyl)acetic acid
To a solution of 2-(4-methoxy-2-phenoxyphenyI)-l-morpholin-4-yIethan-l-thione 10 (0.35 g, 1.02 mmole) in i-PrOH (15 mL) and H2O (15 mL) was added KOH (0.57 g, 10.2 mmole). The reaction was heated at reflux for 18 hr, then was cooled to RT, diluted withH2O, and washed with Et2O. The aqueous layer was acidified to pH ~ 4 with conc. HCland was extracted with CHCI3. The combined extracts were dried over MgSC>4 andconcentrated to give the title compound (0.22 g) as a white solid. This was used without 15 further purification: MS (ES) m/e 259.0 (M + H)+. d) 3-Methoxydibenzo[b,f]oxepin-10-one A solution of 2-(4-methoxy-2-phenoxyphenyl)acetic acid (594 mg, 2.3 mmole) inthionyl chloride (10 mL) was heated at reflux for 30 min, then was concentrated to dryness, 20 and the residue was reconcentrated from toluene. The resulting residue was dissolved indry CH2CI2 (3 mL), and the solution was added dropwise at RT to a suspension of AICI3(673 mg, 5.06 mmole)in dry CH2CI2 (4 mL) in a flame-dried flask under argon. Afterstirring for 2.5 hr, the mixture was diluted with CH2CI2 (10 mL) and washed sequentiallywith 1.0 N NaOH and brine. Drying (MgSO4), concentration, and flash chromatography on 25 silica gel (5% EtOAcZhexanes) gave the title compound (264 mg, 48%) as a light yellowoil: !h NMR (300 MHz, CDCI3) δ 3.80 (s, 3 H), 4.02 (s, 2 H), 6.74 - 8.08 (m, 7 H). e) Ethy 1 (±)-10,11 -dihydro-10-hydroxy-3-methoxydibeiizo[b,f]oxepine-10-acetate
Anhydrous EtOAc (0.94 mL, 9.6 mmole) was added dropwise to a solution of 30 lithium bis(trimethylsilyl)amide (1.0 M in THF, 7 mL, 7 mmole) in dry THF (7 mL) in aflame-dried flask at -78 °C under argon. After 0.5 hr, TMEDA (2.4 mL, 16 mmole) wasadded. After another 5 min, a solution of 3-methoxydibenzo[b,f]oxepin- 10-one (760 mg,3.2 mmole) in THF (2 mL) was added dropwise over 3 min. Additional dry THF (0.4 mL)was used in transfer. The reaction was stirred at -78 °C to -40 °C for 1 hr, then was 35 quenched with saturated NH4CI (10 mL). The mixture was warmed to RT and extractedwith EtOAc. Drying (MgSO4), concentration, and flash chromatography on silica gel (10%EtOÀc/hexanes) gave the title compound as a clear oil: *H NMR (300 MHz, CDCI3) δ 011341 -52- 1.14- 1.2O(t,3 H), 1.21 - 1.30 (m, 1 H), 2.62-2.68 (dd, 1 H), 2.94 - 3.02 (dd, 1 H), 3.24 - 3.30 (dd, 1 H), 3.40 - 3.46 (dd, 1 H), 3.40 - 3.46 (dd, 1 H), 3.78 (s, 3 H), 4.08 - 4.18 (m, 2H), 6.60 - 7.26 (m, 6 H), 7.64 - 7.68 (dd, 1 H). 5 f) Ethyl (±)-10,1 l-dihydro-3-methoxydibenzo[b,f]oxepine-10-acetate
Boron trifluoride etherate (0.48 mL, 3.9 mmole) was added to a solution of ethyl (±)-10,l l-dihydro-10-hydroxy-3-methoxydibenzo[b,f]oxepine-10-acetate (690 mg, 1.95mmole) and triethylsilane (0.62 mL, 3.9 mmole) in dry CH2CI2 at 0 °C under argon. After20 min, the reaction was quenched with 5% NaHCOj, and the mixture was extracted with 10 CH2CI2. Drying (MgSO4) and concentration gave a yellow oil. This was dissolved inabsolute éthanol (20 mL) and 10 % Pd/C (413 mg, 0.39 mmole) was added. The mixturewas hydrogenated for 3 hr at 50 psi on a Parr hydrogénation apparatus. The catalyst wasremoved by filtration through celite®, and the filtrate was concentrated to afford the titlecompound (523 mg, 86%) as a clear oil: 1H NMR (300 MHz, CDCI3) δ 7.18-6.58 (m, 7 15 H), 4.18 - 4.08 (m, 2 H), 3.80 (s, 3 H), 3.80 - 3.74(m, 1 H), 3.40 - 3.30 (dd, 1 H), 2.98 - 2.84 (dd, 1 H), 2.74 - 2.62 (dd, 1 H), 2.60 - 2.52(m, 1 H), 1.32 - 1.20 (t, 3 H). g) Ethyl (±)-10,l l-dihydro-3-hydroxydibenzo[b,f]oxepine-10-acetate A solution of ethyl (±)-10,l l-dihydro-3-methoxydibenzo[b,f]oxepine-10-acetate 20 (523 mg, 1.68 mmole) in CH2CI2 (6.8 mL) was added dropwise to a cold solution of BBrç in CH2CI2 (1.0 M, 6.7 mL, 6.7 mmole) at 0 °C under argon. The reaction was stirred for20 min, then CH3OH (7 mL) was added carefully. The mixture was concentrated and theresidue was flash chromatographed on silica gel (15 - 20% EtOAc/hexanes) to afford thetitle compound (407 mg, 89%) as a pale yellow oil: MS (ES) m/e 299 (M + H)+. 25
Préparation 21
Préparation of 2-f(3-bromo-l-propyl)aminol-4-methylpyridine-N-oxide hydrobromide 30 a) 2-[(3-Bromo-l-propyl)amino]-4-methylpyridine-N-oxide hydrobromide A solution of SOBrç (5.0 mL, 64.5 mmole) in CH2CI2 (20 mL) was added dropwise over 15-20 min to a solution of 2-((3-hydroxy-l-propyl)amino]-4-methylpyridine-N-oxide (10.0 g, 54.87 mmole) in CH2CI2 (100 mL) at 0 °C. The reactionwas warmed to RT and stirred for 2 hr, then Et2Û (200 mL) was added slowly. The 35 solvents were decanted away from the gummy precipitate, and the precipitate was washedwith additional (several times). The resulting brownish-yellow residue solidified on standing in a refrigerator ovemight. This solid was collected and washed with -53- 011341
Et2Û to afford the titie compound (55.07 g) as a yellow soiid. Additionai titie compound(2.05 g) was obtained as white needles by concentration of the combined organic layers.The total yield of titie compound was 17.89 g (96%): MS (ES) m/e 245 and 247 (M + H)+. 5 The following compounds illustrate methods for preparing the biologically active compounds of this invention from intermediate compounds such as described in theforegoing Préparations.
Example 1 10
Préparation of (±)-10.11-dihvdro-3-[4-(pyridin-2-vlamino)-l-butvll-5H- dibenzofa,d1cycloheptene- 10-acetic acid a) 4-(2-Tetrahydropyranyloxy)- 1-tributylstannyl- 1-butyne 15 A solution of n-butyllithium in hexanes (1.6 M, 18.8 mL, 30 mmole) was added in a stream over 2 min to a solution of 2-(3-butynyloxy)tetrahydro-2//-pyran (4.7 mL, 30mmole) in dry THF (60 mL) at 0°C under argon. After 0.5 hr, tributyltin chloride (8.1 mL,30 mmole) was added ail at once, and the reaction was warmed to RT. After 3 hr, thereaction was diluted with hexanes (300 mL) and washed sequentially with H2O (2 x 60 20 mL), 10% KF (2 x 30 mL), and saturated brine (60 mL). Drying (Na2SO4), concentration,and silica gel chromatography (3% EtOAc/hexanes) gave the titie compound (3.58 g, 27%)as a nearly colorless oil: TLC (5% EtOAc/hexanes) Rf 0.37; NMR (400 MHz, CDCI3)Ô 4.66 (narrow t, 1 H), 3.75 - 3.96 (m, 2 H), 3.49 - 3.62 (m, 2 H), 2.56 (app t, 2 H), 1.76 -1.91 (m, 1 H), 1.65 - 1.78 (m, 1 H), 1.42 - 1.65 (m, 10 H), 1.22 - 1.41 (m, 6 H), 0.82 - 1.08 25 (m, 15 H). b) Ethyl (±)-10,ll-dihydro-3-[4-(2-tetrahydropyranyloxy)-l-butyn-l-yl]-5H-dibenzo[a,d]cycloheptene-10-acetate A mixture of ethyl (±)-10,l l-dihydro-3-(trifluoromethanesulfonyloxy)-5H- 30 dibenzo[a,d]cycloheptene-10-acetate (1.34 g, 3.13 mmole), 4-(2-tetrahydropyranyloxy)-l- tributylstannyl-1-butyne (1.66 g, 3.76 mmole), LiCl (398 mg, 9.39 mmole),bis(triphenylphosphine)palladium dichloride (110 mg, 0.094 mmole), and anhydrousdioxane (31 mL) was heated at reflux under argon. After 1.5 hr, the reaction wasconcentrated to remove most of the dioxane, and the residue was taken up in Et2Û (100 35 mL). 10% KF (50 mL) was added and the mixture was stirred briskly for 0.5 hr. Theaqueous layer was removed and the Et2Û layer was filtered through a mixture of celite®and MgSC>4. The filtrate was concentrated and the residue was chromatographed on silica -54- 011341 gel ( 10% EtOAc/hexanes) to afford the title compound (1.12g, 83%) as a pale yellow oil;TLC (20% EtOAc/hexanes) Rf 0.40; !H NMR (400 MHz, CDC13) δ 7.21 - 7.30 (m, 1 H),7.06 -7.20 (m, 5 H), 7.00 (d, J = 7.8 Hz, 1 H), 4.69 (t, J = 3.6 Hz, 1 H), 4.31 (d, J = 15.2 Hz, 1 H), 4.11 - 4.23 (m, 2 H), 3.76 - 3.97 (m, 4 H), 3.59 - 3.68 (m, 1 H), 3.48 - 3.57 (m, 1 H), 5 3.34 (dd, J = 15.2, 4.1 Hz, 1 H), 2.97 (dd, J = 15.2, 9.5 Hz, 1 H), 2.70 (t, J = 7.3 Hz, 2 H), 2.65 (dd, J= 15.7,4.8 Hz, 1 H), 2.51 (dd, J= 15.7, 9.5 Hz, 1 H), 1.78- 1.92 (m, l H), 1.68-1.78 (m, 1 H), 1.44 - 1.68 (m, 4 H), 1.27 (t, J = 7.1 Hz, 3 H); MS (ES) m/e 455 (M + Na)+. c) Ethyl (±)-10,l l-dihydro-3-[4-(2-tetrahydropyranyloxy)-l-butyl]-5H- 10 dibenzo[a,d]cycloheptene- 10-acetate A mixture of ethyl (±)-10,l l-dihydro-3-[4-(2-tetrahydropyranyloxy)-l-butyn-l-yl]-5H-dibenzo[a,d]cycloheptenë-10-acetate (1.2 g, 2.77 mmole), 10% Pd/C (0.3 g, 0.28mmole), and EtOAc (28 mL) was shaken at RT under hydrogen (50 psi) on a Parrapparatus. After 3 hr, the reaction was fîltered through celite® and the filtrate was 15 concentrated. Silica gel chromatography ( 10% EtOAc/hexanes) gave the title compound(1.06 g, 88%) as a colorless oil: TLC (20% EtOAc/hexanes) Rf 0.51; ^H NMR (400 MHz,CDCI3) δ 7.05 - 7.20 (m, 4 H), 6.92 - 7.03 (m, 3 H), 4.53 - 4.60 (m, 1 H), 4.34 (d, J = 15.1Hz, 1 H), 4.12 - 4.26 (m, 2 H), 3.80 - 3.90 (m, 3 H), 3.71 - 3.80 (m, 1 H), 3.44 - 3.53 (m, 1H), 3.35 - 3.44 (m, 1 H), 3.33 (dd, J = 15.1, 4.1 Hz, 1 H), 2.95 (dd, J = 15.1, 9.4 Hz, 1 H), 20 2.65 (dd, J = 15.5,4.9 Hz, 1 H), 2.49 - 2.61 (m, 3 H), 1.77 - 1.90 (m, 1 H), 1.45 - 1.77 (m, 9 H), 1.27 (t, J = 7.1 Hz, 3 H); MS (ES) m/e 459 (M + Na)+. d) Ethyl (±)-10,l l-dihydro-3-(4-hydroxy-l-butyl)-5H-dibenzo[a,d]cycloheptene-lO-acetate 25 A solution of ethyl (±)Ί0,1 l-dihydro-3-[4-(2-tetrahydropyranyloxy)-l-butyl]-5H- dibenzo[a,d]cycloheptene-10-acetate (456.0 mg, 1.04 mmole) and p-toluenesulfonic acidmonohydrate (60 mg, 0.31 mmole) in absolute EtOH (10 mL) was stirred at RT. After 2 hr,the reaction was quenched with 5 % NaHCC>3 (1 mL) and concentrated to remove theEtOH. The residue was diluted with H2O (2 mL) and extracted with CH2CI2. Drying 30 (MgSO4), concentration, and silica gel chromatography (1:1 EtOAc/hexanes) gave the titlecompound (342.4 mg, 93%) as a colorless oil: TLC (1:1 EtOAc/hexanes) Rf 0.49; ^HNMR (250 MHz, CDCI3) δ 6.85 - 7.25 (m, 7 H), 4.34 (d, J = 15.1 Hz, 1 H), 4.08 - 4.30 (m, 2 H), 3.75 - 3.95 (m, 2 H), 3.53 - 3.72 (m, 2 H), 3.33 (dd, J = 15.1, 4.1 Hz, 1 H), 2.95 (dd, J= 15.1, 9.4 Hz, 1 H), 2.40 - 2.75 (m, 4 H), 1.45 - 1.80 (m, 4 H), 1.27 (t, J = 7.1 Hz, 3 H); 35 MS (ES) m/e 353 (M + H)+. -55- 5 011341 e) Ethy! (±)-10,l !-dihydro-3-[4-(N-phtha!idim;do)-!-bÎ;ty!]-5K-dibenzoLa,d]cycÎohepicne- 10-acetate
Diethyl azodicarboxylate (0.2 mL, 1.26 mmole) was added dropwise to a solutionof ethyl (±)-10,l l-dihydro-3-(4-hydroxy-l-butyl)-5H-dibenzo[a,d]cycloheptene-10-acetate(0.37 g, 1.05 mmole), triphenylphosphine (0.33 g, 1.26 mmole), and phthalimide (0.19 g,1.26 mmole) in anhydrous THF (10 mL) at RT under argon. After 23 hr, the reaction wasconcentrated on tbe rotavap. Silica gel chromatography (30% EtOAc/hexanes) gave thetitle compound (0.35 g, 70 %) as a colorless oil: MS (ES) m/e 504.3 (M + Na)+. 10 f) Ethyl (±)-10,11 -dihydro-3-(4-amino-1 -butyl)-5H-dibenzo[a,d]cycloheptene-10-acetateHydrazine monohydrate (0.11 g, 2.18 mmole) was added to a solution of ethyl (±)- 10,11 -dihydro-3-[4-(N-phthalidimido)-1 -butyl]-5H-dibenzo[a,d]cycloheptene-10-acetate(0.35 g, 0.73 mmole) in absolute EtOH (10 mL) and toluene (2 mL) at RT. The reactionwas stirred at RT for 17 hr, then was fïltered, and the filter pad was washed with toluene. 15 Concentration on the rotavap gave the title compound (0.23 g, 90%) as a colorless solid:MS (ES) m/e 352.3 (M + H)+. g) Ethyl (±)-10,ll-dihydro-3-[4-(l-oxopyridin-2-ylamino)-l-butyl]-5H-dibenzo[a,d]cycloheptene-10-acetate 20 A mixture of 2-chloropyridine-N-oxide hydrochloride (0.31 g, 1.88 mmole), ethyl (±)-10,l l-dihydro-3-(4-amino-l-butyl)-5H-dibenzo[a,d]cycloheptene-l 0-acetate (0.22 g,0.63 mmole), and NaHCÛ3 (0.26 g, 3.13 mmole) in zcrr-amyl alcohol (6 mL) was heated atreflux for 21 hr. The reaction mixture was diluted with CH2CI2 (100 mL) and fïltered, andthe filtrate was concentrated on the rotavap. Silica gel chromatography (1:9:5 25 MeOH/CH2Cl2/EtOAc) gave the title compound (82 mg, 30%) as a yellow oil: MS (ES)m/e 445.2 (M + H)+. h) Ethyl (±)-10,ll-dihydro-3-[4-(pyridin-2-ylamino)-l-butyl]-5H-dibenzo[a,d]cycloheptene-10-acetate 30 A mixture of ethyl (±)-10,l l-dihydro-3-[4-(l-oxopyridin-2-ylamino)-l-butyl]-5H- dibenzo[a,d]cycloheptene-10-acetate (0.07 g, 0.16 mmole), 10% Pd/C (0.08 g, 0.075mmole), cyclohexene (0.16 mL, 1.6 mmole), and isopropanol (4 mL) was heated at refluxunder argon for 14 hr, then the catalyst was removed by filtration through celite®. Thefilter pad was washed with isopropanol and MeOH, and the filtrate was concentrated on the 35 rotavap to give the title compound (0.046g, 69%) as a clear oil: MS (ES) m/e 429.3 (M +H)+. -56- 011341 i) Ethyl (±)-10,l l-dihydro-3-[4-(pyridin-2-ylamino)-l-butyl]-5H-dibenzo[a,d]cycloheptene-10-acetic acid A mixture of ethyl (±)-10,l l-dihydro-3-[4-(pyridin-2-ylamino)-l-butyl]-5H-dibenzo[a,d]cycloheptene-l 0-acetate (46 mg, 0.11 mmole) and 1.0 N LiOH (0.66 mL, 0.66 5 mmole) in THF (3 mL) and H2O (3 mL) was stirred at RT. After 24 hr, the reactionmixture was concentrated on the rotavap, and the residue was diluted with H2O (5 mL).
The solution was cooled in an ice bath, and 1.0 N AcOH was added slowly to give a whiteprecipitate. Chromatography on C-18 YMC (45% CH3CN/H2O containing 0.1% TFA)gave the title compound ( 13 mg, 21 %) as a white solid: MS (ES) m/e 401.3 (M + H)+. 10 Anal. Calcd for C26H28N2O2 · 0.75 H2O · 1.5 CF3CO2H: C, 59.54; H, 5.31 ; N, 4.72.Found: C, 59.69; H, 5.31 ; N, 4.72.
Example 2 15 Préparation of (±)-10,l l-dihvdro-3-f3-(4-ethoxvpvridin-2-ylamino)-l-propyloxvl-5H- dibenzoia,dlcycloheptene- 10-acetic acid a) Ethyl (±)-10,11 -dihydro-3-[3-(4-ethoxy-1 -oxopyridin-2-ylamino)-1 -propyloxy]-5H-dibenzo[a,d]cycloheptene-10-acetate 20 Ethyl (±)-10,1 l-dihydro-3-[3-(4-nitro-l-oxopyridin-2-ylamino)-l-propyloxy]-5H- dibenzo[a,d]cycloheptene-10-acetate (0.67 g, 1.36 mmole), 1.0 M NaOEt in éthanol (6.8mL, 6.8 mmole), and absolute éthanol (6.8 mL) were combined, and the mixture waswarmed in an oil bath preset at 70°C. A dark solution was produced, which was warmedfor 10 min, then the oil bath was removed, and the solution was allowed to stir for an 25 additional 5-7 min without extemal heating. The resulting solution was cooled in ice, andthe reaction was quenched with glacial acetic acid (0.47 mL, 8.2 mmole). The mixture wasconcentrated and the residue was partitioned between CH2CI2 (10 mL) and half-saturatedNH4CI (10 mL). The layers were separated and the aqueous layer was extracted withCH2CI2 (2x10 mL). The combined organic layers were dried (MgSC>4) and concentrated, 30 and the residue was reconcentrated from toluene to leave a reddish-orange oil. Silica gelchromatography (5% MeOH/CHCl3) gave the title compound (601.1 mg, 90%) as a yellowoil: TLC (5% MeOH/CHCl3) Rf 0.36; 1h NMR (25QMHz, CDC13) δ 7.95 (d, J = 7.1 Hz, 1H), 6.88 - 7.30 (m, 6 H), 6.77 (d, J = 2.6 Hz, 1 H), 6.67 (dd, J = 8.2, 2.6 Hz, 1 H), 5.95 - 6.20 (m, 2 H), 4.28 (d, J = 15.0 Hz, 1 H), 4.18 (q, J = 7.2 Hz, 2 H), 4.04 (t, J = 5.6 Hz, 2 H), 35 3.65 - 4.00 (m, 4 H), 3.46 (q, J = 6.5 Hz, 2 H), 3.30 (dd, J = 15.0, 4.2 Hz, 1 H), 2.93 (dd, J = 15.0, 9.2 Hz, 1 H), 2.65 (dd, J = 15.6, 5.0 Hz, 1 H), 2.52 (dd, J = 15.6, 9.4 Hz, 1 H), 1.95 -2.25 (m, 2 H), 1.10- 1.45 (m, 6 H); MS (ES) m/e 491 (M + H)+. - 57 - 011341 b) Ethyl (±)-10,l l-dihydro-3-[3-(4-ethoxypyridin-2-ylamino)-l-propyloxy]-5H-dibenzo[a,d]cycloheptene-10-acetate A mixture of ethyl (±)-10,l l-dihydro-3-[3-(4-ethoxy-l-oxopyridin-2-ylamino)-I- 5 propyloxy]-5H-dibenzo[a,d]cycloheptene-l0-acetate (601.1 mg, 1.23 mmole), cyclohexene(1.2 mL, 12.3 mmole), 10% Pd/C (130 mg, 0.012 mmole), and absolute éthanol (12.3 mL)was heated at reflux under argon. After 23.5 hr, the reaction was hot-filtered throughcelite® and the filter pad was washed with éthanol. The filtrate was concentrated and theresidue was reconcentrated from toluene. Silica gel chromatography (5% MeOH in T. 1 10 EtOAc/CHClj) gave the title compound (528.1 mg, 90%) as a light yellow oil; TLC (10%MeOH in EtOAc/CHCl3) Rf 0.67; JH NMR (400 MHz, CDC13) δ 7.89 (d, J = 5.8 Hz, 1 H),7.05 - 7.18 (m, 4 H), 6.99 (d, J = 8.2 Hz, 1 H), 6.77 (d, J = 2.6 Hz, 1 H), 6.66 (dd, J = 8.2, 2.6 Hz, 1 H), 6.17 (dd, J = 5.8, 2.1 Hz, 1 H), 5.86 (d, J = 2.1 Hz, 1 H), 4.73 (br t, 1 H), 4.28(d, J = 14.9 Hz, 1 H), 4.11 - 4.25 (m, 2 H), 4.04 (t, J = 5.9 Hz, 2 H), 3.98 (q, J = 7.0 Hz, 2 15 H), 3.83 (d, J = 14.9 Hz, 1 H), 3.76 - 3.85 (m, 1 H), 3.43 (q, J = 6.4 Hz, 2 H), 3.30 (dd, J = 15.0,4.1 Hz, 1 H), 2.93 (dd, J = 15.0, 9.2 Hz, 1 H), 2.64 (dd, J = 15.6, 4.8 Hz, 1 H), 2.52(dd, J = 15.6, 9.5 Hz, 1 H), 2.01 - 2.11 (m, 2 H), 1.37 (t, J = 7.0 Hz, 3 H), 1.27 (t, J = 7.0Hz, 3 H); MS (ES) m/e 475 (M + H)+. 20 c) (±)-10,l l-Dihydro-3-[3-(4-ethoxypyridin-2-ylamino)-l-propyloxy]-5H-dibenzo(a,d]cycloheptene-10-acetic acid 1.0 N NaOH (1.7 mL, 1.7 mmole) was added dropwise to a solution of ethyl (±)- 10,1 l-dihydro-3-[3-(4-ethoxypyridin-2-ylamino)-l-propyloxy]-5H-dibenzo[a,d]cycloheptene-10-acetate (528.1 mg, 1.11 mmole) in absolute éthanol (11 mL) 25 at RT, and the solution was warmed in an oil bath preset at 45°C. After 20 hr, the reactionwas concentrated, and the residue was reconcentrated from H2O. The resulting residue wasdissolved in H2O (10 mL) and the solution was filtered. The pH was adjusted to 7 with 1.0N HCl, and the mixture was stirred briskly to convert thé initially-formed gummyprecipitate into a solid. Trituration with a glass rod and a spatula aided in this 30 transformation. The pH of the resulting mixture was readjusted to 7, and the solid wascollected and washed with plenty of H2O. The filtrate was concentrated and the residuewas dissolved in H2O with the aid of a little 1.0 N NaOH. The pH was adjusted to 7 toafford a small second crop. The crops were combined and dried in vacuum (40 - 50°C) toafford the title compound (453.7 mg, 82%) as an off-white solid; HPLC (Hamilton PRP- 35 1®, 45% CH3CN/H2O containing 0.1 % TFA) k’ = 1.32; 1H NMR (400 MHz, DMSO-d6) δ 7.78 (d, J = 6.6 Hz, 1 H), 7.35 - 7.65 (m, 1 H), 7.02 - 7.22 (m, 4 H), 6.97 (d, J = 8.3 Hz, 1H), 6.82 (d, J = 2.4 Hz, 1 H), 6.68 (dd, J = 8.3, 2.4 Hz, 1 H), 6.29 (dd, 1 H), 6.15 (narrow d, -58- 011341 I H), 4.20 (d, J = 14.6 Hz, 1 H), 3.93 - 4.12 (m, 4 H), 3.89 (d, J = 14.6 Hz, 1 H), 3.60 - 3.71(m, 1 H), 3.30 - 3.50 (m, 2 H), 3.20 (dd, J = 15.1,4.1 Hz, 1 H), 2.83 (dd, J = 15.1, 10.1 Hz,
1 H), 2.60 (dd, J = 16.0, 5.3 Hz, 1 H), 2.48 (dd, J = 16.0, 8.9 Hz, 1 H, partially obscured byresidual solvent signal), 1.90 - 2.05 (m, 2 H), 1.30 (t, J = 6.9 Hz, 3 H); MS (ES) m/e 447 (M 5 + H)+. Anal. Calcd for C27H30N2O4 · 1.5 HCl: C, 64.70; H, 6.33; N, 5.59. Found: C, 64.53; H, 6.14; N, 5.31.
Example 3 10 Préparation of (ί)-ΙΟ,Ι l-dihvdro-3-f2-i2-(ethvlamino)thiazol-4-yll-l-ethoxyl-5H- dibenzofa,d]cvcloheptene- 10-acetic acid a) Ethyl (±)-10,1 l-dihydro-3-[2-[2-(ethylamino)thiazol-4-yl]-l-ethoxy]-5H-dibenzo[a,d]cycloheptene-10-acetate 15 A solution of 2-(ethylamino)-4-thiazoleethanol (0.33 g, 1.9 mmole) and diethyl azodicarboxylate (0.30 mL, 1.9 mmole) in anhydrous DMF (5 mL) was added dropwiseover 5 min to a solution of ethyl (±)-10,l l-dihydro-3-hydroxy-5H- dibenzo[a,d]cycloheptene-10-acetate (296.4 mg, 1 mmole) and triphenylphosphine (525mg, 2 mmole) in anhydrous DMF (5 mL) at RT. The reaction was kept cool in a RT water 20 bath during the addition. After 16 hr, the reaction was concentrated and the residue wasreconcentrated from xylenes (2 x). Silica gel chromatography (20% EtOAc/hexanes) gavethe title compound (145.0 mg, 32%) as a yellow oil: TLC (1:1 EtOAc/hexanes) Rf 0.60; ’H NMR (250 MHz, CDCI3) δ 7.00 - 7.30 (m, 4 H), 6.98 (d, J = 8.2 Hz, 1 H), 6.77 (d, J = 2.6 Hz, 1 H), 6.68 (dd, J = 8.2, 2.6 Hz, 1 H), 6.21 (s, 1 H), 5.00 - 5.25 (m, 1 H), 4.04 - 4.38 25 (m, 5 H), 3.81 (d, J = 15.1 Hz, 1 H), 3.70 - 3.90 (m, 1 H), 3.13 - 3.40 (m, 3 H), 2.99 (t, J = 6.7 Hz, 2 H), 2.92 (dd, J = 14.9, 9.3 Hz, 1 H), 2.64 (dd, J = 15.6, 5.0 Hz, 1 H), 2.51 (dd, J =15.6, 9.3 Hz, 1 H), 1.27 (t, J = 7.2 Hz, 3 H); MS (ES) m/e 451 (M + H)+. b) (±)-10,1 l-Dihydro-3-[2-[2-(ethylamino)thiazol-4-yl]-1 -ethoxy]-5H- 30 dibenzo[a,d]cycloheptene-10-acetic acid 1.0 N LiOH (0.32 mL, 0.32 mmole) was added dropwise to a solution of ethyl (±)- 10,1 l-dihydro-3-[2-[2-(ethylamino)thiazol-4-yl]-l-ethoxy]-5H-dibenzo[a,d]cycloheptene-10-acetatè ( 145.0 mg, 0.32 mmole) in THF (2.4 mL) and H2O (0.48 mL) at 0°C. Theresulting pinkish-orange two-phase mixture was stirred at 0°C for 10 min, during which 35 time the color faded to orangish-yellow, then was warmed to RT. After 1.5 hr, a little moreH2O (5 drops) was added, and the reaction was stirred for 42 hr, then was cooled to 0°Cand neutralized with TFA (0.025 mL). The THF was removed on the rotavap, and the 011341 -59- 5 resulting oily residue was düuted with 0.1% TFA/CH3CN to give a homogeneous solution.ODS chromatography (gradient: 40% CH3CN/H2O containing 0.1% TFA, then 45%CH3CN/H2O containing 0.1% TFA) gave fractions containing the title compound. Thesewere pooled, and the CH3CN was removed on the rotavap. The resulting aqueous mixturewas made basic at 0°C to afford a homogeneous solution. Careful acidification to pH 4 - 5with 1.0 N HCI gave a solid precipitate, which was collected, washed with plenty of H2O,and dried to afford the title compound (80.9 mg, 51%) as an off-white solid: HPLC(Hamilton PRP-1®, 45% CH3CN/H2O containing 0.1% TFA) k’ = 0.89; NMR (400MHz, CD3OD) δ 7.02 - 7.18 (m, 4 H), 7.00 (d, J = 8.3 Hz, 1 H), 6.79 (d, J = 2.6 Hz, 1 H), 10 15 6.68 (dd, J = 8.3, 2.6 Hz, 1 H), 6.45 (s, 1 H), 4.26 (d, J = 14.9 Hz, 1 H), 4.20 (t, J = 6.4 Hz,2 H), 3.87 (d, J = 14.9 Hz, 1 H), 3.68 - 3.80 (m, 1 H), 3.34 (q, J = 7.3 Hz, 2 H, partiallyobscured by residual solvent signal), 3.30 (dd, 1 H, obscured by residual solvent signal),2.99 (t, J = 6.4 Hz, 2 H), 2.92 (dd, J = 15.0, 9.4 Hz, 1 H), 2.62 (dd, J = 15.9, 5.0 Hz, 1 H),2.47 (dd, J = 15.9, 9.3 Hz, 1 H), 1.27 (t, J = 7.3 Hz, 3 H); MS (ES) m/e 423 (M + H)+.Anal. Calcd for C24H26N2O3S · 0.67 CF3CO2H: C, 61.03; H, 5.39; N, 5.62. Found: C, 61.21; H, 5.36; N, 5.60.
Example 4 20 Préparation of (S)-10,l l-dihvdro-3-i3-(pyridin-2-vlamino)-l-propyloxvl-5H- dibenzoia,dlcycloheptene-10-acetic acid a) Ethyl (S)-10,l l-dihydro-3-[3-(l-oxopyridin-2-ylamino>l-propyloxy]-5H-dibenzo[a,d]cycloheptene-10-acetate 25 To a stirred solution of ethyl (S)-10,11 -dihydro-3-hydroxy-5H- dibenzo[a,d]cycloheptene-10-acetate (35 g, 118 mmol) in dry THF (1.1 L) and dry DMF(600 mL) under argon were added 2-[(3-hydroxy-l-propyl)amino]pyridine-N-oxide (29.4 g,175 mmol) and triphenylphosphine (45.9 g, 175 mmol). After ail solids had completelydissolved (~1 h), the reaction was cooled to 0 °C in an ice bath and diisopropyl
30 azodicarboxylate (36.4 mL, 95%, 175 mmol) was added via syringe. The reaction wasallowed to warm slowly to RT and was stirred for 18 h. Concentration and flashchromatography on silica gel (95:5 CHCl3/MeOH) followed by a second purification byflash chromatography on silica gel (80:20:5 CHCl3/EtOAc/EtOH) gave the title compound(37.66 g, 71%) as a pale yellow solid foam: ^H NMR (400 MHz, DMSO-dg) δ 8.08 (dd, J 35 = 6.3, 1.1 Hz, 1H), 7.29 (t, 1H), 7.19-7.06 (m, 5H), 6.97 (d, J = 8.3 Hz, 1H), 6.84 (d, J = 2.5, 1H), 6.79 (dd, J = 8.5, 1.6 Hz, 1H), 6.69 (dd, J = 8.3, 2.6 Hz, 1H), 6.57 (m, 1H), 4.17(d, J = 14.7 Hz, 1H), 4.13-4.07 (m, 2H), 4.00 (t, 2H), 3.91 (d, J = 14.7 Hz, 1H), 3.66 (m, 011341 -60- 1H), 3.39 (t, 2H), 3.19 (dd, J = 15.1,4.5 Hz, 1H), 2.85 (dd, J = 15.1, 10.0 Hz, 1H),2.65 (dd, J = 15.8,5.4 Hz, 1H), 1.99 (m, 2H), 1.18 (t, 3H); MS (ES) m/e 447.3 (M + H)+. b) Ethyl (S)-10,l l-dihydro-3-[3-(pyridin-2-ylamino)-l-propyloxy]-5H- 5 dibenzo[a,d]cycloheptene- 10-acetate
To a stirred solution of ethyl (S)-10,1 l-dihydro-3-[3-(l-oxopyridin-2-ylamino)-l-propyloxy]-5H-dibenzo[a,d]cycloheptene-10-acetate (37.66 g, 84 mmol) in isopropanol(700 mL) were added 10% palladium on activated carbon (18 g, 16.9 mmol, carefully pre-wetted in isopropanol under argon) and cyclohexene (85 mL, 839 mmol). The reaction was 10 then heated to reflux under argon in an oil bath set at 90 °C . After 6 h an additionalamount of 10% palladium on activated carbon (18 g, 84 mmol, carefully pre-wetted inisopropanol under argon) and cyclohexene (85 mL, 839 mmol) were added. After anadditional 18 h the reaction was hot-filtered through celite®, and the filter pad was washedwith 1:1 MeOH/CHCl3 (600 mL). The filtrate was concentrated under vacuum and the 15 residue was purified by flash chromatography on silica gel (95:5 CHC^/MeOH) to give thetitle compound (29.2 g, 81%) as a pale yellow oil: ^H NMR (400 MHz, DMSO-d^) Ô 7.94(dd, J = 5.4, 1.9 Hz, 1H), 7.35-7.31 (m, 1H), 7.18 (d, J = 7.2 Hz, 1 H), 7.14-7.06 (m, 3H),6.97 (d, J = 8.3 Hz, 1H), 6.83 (d, J = 2.6, 1H), 6.68 (dd, J = 8.3, 2.6 Hz, 1H), 6.54 (t, 1H),6.44 (m, 2H), 4.17 (d, J = 14.6 Hz, 1H), 4.13-4.02 (m, 2H), 4.00 (t, 2H), 3.91 (d, J = 14.7 20 Hz, 1H), 3.66 (m, 1H), 3.35 (m, 2H), 3.19 (dd, J = 15.1, 4.4 Hz, 1H), 2.86 (dd, J = 15.1, 10.1 Hz, 1H), 2.65 (dd, J = 15.8, 5.4 Hz, 1H), 2.55 (dd, J = 15.8, 8.7 Hz, 1H), 1.93 (m, 2H),1.18 (t, 3H); MS (ES) m/e 431.4 (M + H)+. c) (S)-10,l l-Dihydro-3-[3-(pyridin-2-ylamino)-l-propyloxy]-5H- 25 dibenzo[a,d]cycloheptene-10-acetic acid
To a stirred solution of ethyl (S)-10,1 l-dihydro-3-[3-(pyridin-2-ylamino)-l-propyloxy]-5H-dibenzo[a,d]cycloheptene-10-acetate (29.20 g, 68 mmol) in dioxane (350mL) was added aqueous 1.0 N NaOH (110 mL, 110 mmol). The cloudy reaction wasstirred at 50 °C in an oil bath for 24 h, then the resulting homogeneous solution was 30 neutralized with aqueous 1.0 N HCl (110 mL, 110 mmol). The solution was concentratedto near dryness by rotary évaporation to precipitate out the product. The supematant wasdecanted off and the remaining gummy solid was drièd under vacuum and redissolved in1:1 methanol/CHC^. The clear solution was then reconcentrated by rotary évaporation andthoroughly dried under vacuum. The remaining solid was triturated with a small volume of 35 water, filtered and dried under vacuum to give the title compound (26.85 g, 94%) as an off-white powder: HPLC (Hamilton PRP-1®, 35% CH3CN/H2O containing 0.1 % TFA) k’ =2.88; !h NMR (400 MHz, DMSO-d6) δ 7.94 (dd,J = 4.7, 1.6 Hz, 1H), 7.38 (m, 1H), 7.18 -61 - 011341 (d, J = 7.3 Hz, 1H), 7.14 (d, J = 3.9 Hz, 2H), 7.08 (m, IH), 6.97 (d, i = 8.4 Hz, iH), 6.83 (d,J = 8.6 Hz, 1H), 6.78 (br s, 1H), 6.68 (dd, J = 8.3, 2.6 Hz, 1H), 6.50 (d, J = 8.3 Hz, 1H), 6.47 (dd, 1H), 4.20 (d, J = 14.6 Hz, 1H), 4.00 (t, 2 H), 3.88 (d, J = 14.6 Hz, 1H), 3.67 (m, 1 H), 3.37 (m, 1 H), 3.20 (dd, J = 15.2, 4.4 Hz, 1H), 2.83 (dd, J = 15.2, 10.1 Hz, 1H), 2.60(dd, J = 15.9, 5.3 Hz, 1H), 2.50 (dd, 1H), 1.95 (m, 2 H); MS (ES) m/e 403.3 (M + H)+.Anal. Calcd for C25H26N2O3 · H2O: C, 71.41; H, 6.71; N, 6.66. Found; C, 71.21; H,6.53; N, 6.54.
Example 5 10
Préparation of (±)-10,l l-dihydro-3-i2-(6-aminopyridin-2-yl)-I-ethoxyl-5H- dibenzoia,d1cycloheptene-10-acetic acid a) Ethyl (±)-10,l l-dihydro-3-[2-(6-aminopyridin-2-yl)-l-ethoxy]-5H-15 dibenzo[a,d]cycloheptene-10-acetate A solution of 6-amino-2-pyridylethanol (0.23 g, 1.68 mmole) and diethylazodicarboxylate (0.26 mL, 1.68 mmole) in anhydrous DMF (5 mL) was added dropwise toa solution of ethyl (±)-10,l l-dihydro-3-hydroxy-5H-dibenzo[a,d]cycloheptene-10-acetateand triphenylphosphine (0.48 g, 1.82 mmole) in anhydrous DMF (5 mL) at RT. After 1 hr, 20 the reaction was concentrated and the residue was purified by flash chromatography onsilica gel (1:1 EtOAc/hexanes) to afford the title compound (0.030 g): MS (ES) m/e 417(M + H)+. b) (±)-10,ll-Dihydro-3-[2-(6-aminopyridin-2-yl)-l-ethoxy]-5H-dibenzo[a,d]cycloheptene- 25 10-acetic acid A solution of ethyl (±)-10,1 l-dihydro-3-[2-(6-aminopyridin-2-yl)-l-ethoxy]-5H-dibenzo[a,d]cycloheptene-10-acetate (0.030 g, 0.072 mmole) and 1.0 N NaOH (0.14 mL,0.14 mmole) in MeOH (2 mL) was stirred at RT ovemight, then was concentrated. Theresidue was dissolved in H2O and the pH of the solution was adjusted to 7 with 1.0 N HCl. 30 Concentration and chromatography on a C-18 Bond Elute column (10:9:1 CH3CN/H2O/TFA) gave the title compound (0.013 g): MS (ES) m/e 389 (M + H)+. 011341 -62-
Example 6
Préparation of (R)-10,l l-dihvdro-3-[3-(pvridin-2-ylamino)-l-propyloxvl-5H- dibenzofa.dlcycloheptene-10-acetic acid 5 a) Ethyl (R)-10,l l-dihydro-3-[3-(l-oxopyridin-2-ylamino)-l-propyloxy]-5H-dibenzo[a,d]cycloheptene-lO-acetate A solution of 2-[(3-hydroxy-l-propyl)amino]pyridine-N-oxide (0.70 g, 4 mmole)and diethyl azodicarboxylate (0.65 mL, 4 mmole) in anhydrous DMF (20 mL) was added 10 dropwise over 10 min to a solution of ethyl (R)-10,ll-dihydro-3-hydroxy-5H- dibenzo[a,d]cycloheptene-l 0-acetate (0.45 g, 2 mmole) and triphenylphosphine (1.2 g, 4mmole) in anhydrous DMF (8 mL) at RT under argon. After 23.5 hr, the reaction wasconcentrated on the rotavap, and the residue was reconcentrated from xylenes to removeresidual DMF. Silica gel chromatography (1-4% CH3OH/CH2CI2) gave the title 15 compound (0.50 g, 74 %) as a yellow oil: MS (ES) m/e 447 (M + H)+. b) Ethyl (R)-10,l l-dihydro-3-[3-(pyridin-2-ylamino)-l-propyloxy]-5H-dibenzo[a,d]cycloheptene-10-acetate A mixture of ethyl (R)-10,1 l-dihydro-3-[3-(l-oxopyridin-2-ylamino)-l- 20 propyloxy]-5H-dibenzo[a,d]cycloheptene-l 0-acetate (0.5 g, 1 mmole), 10% Pd/C (0.25 g, 0.2 mmole), cyclohexene (2 mL, 20 mmole), and isopropanol (10 mL) was heated at refluxfor 18 hr, then the catalyst was removed by filtration through celite®. Silica gelchromatography (0.5-2% CH3OH/CH2CI2) gave the title compound (0.4 g, 83%) as a lightyellow oil: MS (ES) m/e 431 (M + H)+. 25 c) (R)-10,1 l-Dihydro-3-[3-(pyridin-2-ylamino)-1 -propyloxy]-5H-dibenzo[a,d]cycloheptene- 10-acetic acid A mixture of ethyl (R)-10,l l-dihydro-3-[3-(pyridin-2-ylamino)-l-propyloxy]-5H-dibenzo[a,d]cycloheptene-l 0-acetate (0.4 g, .93 mmole) and 1.0 N NaOH (1.1 mL, 1.1
30 mmole) in absolute EtOH (10 mL) was warmed in an oil bath set at 50°C. After 18 hr, thereaction was concentrated on the rotavap and the residue was dissolved in H2O. Theaqueous solution was adjusted to pH 4 with 3 N HCl, and the solid precipitate was collectedand washed with H2O. The material was dried in high vacuum at 40°C to afford the titlecompound (0.36 g, 96%) as a nearly colorless solid: [ajg +50.8° (c = 0.12, CH3OH); MS 35 (ES) m/e 403 (M + H)+. Anal. Calcd for C25H26N2O3 · 0.5 H2O: C, 72.97; H, 6.61 ; N,6.80. Found: C, 73.09; H, 6.38; N, 6.58. -63- 011341
Exampie 7
Préparation of (S)-10,l l-dihydro-3-f2-[6-(methylamino)pvridin-2-yll-l-ethoxvl-5H- dibenzofa.dlcycloheptene-10-acetic acid5 a) Ethyl (S)-10,1 l-dihydro-3-[2-[6-(methylamino)pyridin-2-yl]-l-ethoxy]-5H-dibenzo(a,d]cycloheptene-l 0-acetate
According to the procedure of Example 6 (a), except substituting 6-(methylamino)-2-pyridylethanol for the 2-[(3-hydroxy-l-propyl)amino]pyridine-N-oxide, and ethyl (S)- 10 10,1 l-dihydro-3-hydroxy-5H-dibenzo[a,d]cycloheptene-10-acetate for ethyl (R)-10,l 1- dihydro-3-hydroxy-5H-dibenzo[a,d]cycloheptene-10-acetate, the title compound wasobtained as colorless oil following silica gel chromatography (0.2 - 2% MeOH/CH2Cl2):MS (ES) 431.2 (M + H)+. 15 b) (S)-10,1 l-Dihydro-3-[2-[6-(methylamino)pyridin-2-yl]-l-ethoxy]-5H-dibenzo[a,d]cycloheptene-10-acetic acid
Ethyl (S)-10,1 l-dihydro-3-[2-[6-(methylamino)pyridin-2-yl]-l-ethoxy]-5H-dibenzo[a,d]cycloheptene-10-acetate (80 mg, 0.18 mmol) was dissolved in THF (4 mL),and a solution of LiOH · H2O (35 mg, 0.84 mmol) in H2O (4 mL)was added. The solution 20 was stirred at RT for 72 hr, then was diluted with ether (10 mL). The supematant was decanted and the solid was suspended in H2O. Careful acidification to pH 4 with 3 N HClgave the title compound as a white solid: MS(ES) 403 (M + H)+. Anal. Calcd forc25h26n2°3 ’ °·75 h20: c« 72.18; H, 6.66; N, 6.73. Found: C, 72.44; H, 6.52; N, 6.71. 25 Example 8
Préparation of (±)-10,l l-dihydro-3-i3-(3,4.5,6-tetrahvdropyrimidin-2-ylamino)-l- propyloxvl-5H-dibenzora,d1cycloheptene-10-acetic acid 30 * * * * 35 * 30 a) Ethyl (±)-10,1 l-dihydro-3-[3-(4-nitrobenzyloxycarbonyl)amino-l-propyloxy]-5H- dibenzo[a,d]cycloheptene-10-acetate
According to the procedure of Example 6 (a), except substituting 3-(4- nitrobenzyloxycarbonylamino)-l-propanol for the 2-[(3-hydroxy-l-propyl)amino]pyridine- N-oxide, and ethyl (±)-10,ll-dihydro-3-hydroxy-5H-dibenzo[a,d]cycloheptene-l 0-acetate 35 for ethyl (R)-10,l l-dihydro-3-hydroxy-5H-dibenzo[a,d]cycloheptene-10-acetate, the title compound was obtained as amber oil: MS (ES) 533.3 (M + H)+. 011341 -64- b) Ethyl (±)-10,l l-dihydro-3-(3-amino-l-propyloxy)-5H-dibenzo[a,d]cycloheptene-10-acetate A mixture of ethyl (±)-10,l l-dihydro-3-[3-(4-nitrobenzyloxycarbonylamino)-l-propyloxy]-5H-dibenzo[a,d]cycloheptene-10-acetate (1.6 g, 3 mmol), 10% palladium on 5 charcoal (0.8 g, 1 mmol), and éthanol (50 mL) was shaken under H2 (48 psi) for 3 hr, thenthe catalyst was removed by filtration through celite®. The filtrate was concentrated togive the title compound (1.2 g, 100%) as a yellow oil: MS (ES) 348.2 (M + H)+. c) Ethyl (±)-10,l l-dihydro-3-[3-(pyrimidin-2-ylamino)-l-propyloxy ]-5H- 10 dibenzo[a,d]cycloheptene-10-acetate A mixture of ethyl (±)-10,1 1-dihydro-3-(3-amino-l-propyloxy )-5H-dibenzo[a,d]cycloheptene-10'-acetate (0.4 g, 1 mmole), sodium bicarbonate (0.5 g, 6mmole), 2-bromopyrimidine (0.34 g, 2 mmole) and éthanol (10 mL) was heated at refluxunder argon for 18 hr. The solution was then decanted and concentrated. The residue was 15 purified by chromatography on silica gel (0.2 - 2% MeOH/Cl^Clj) to give the titlecompound (0.17 g, 34%) as a pale yellow oil: MS (ES) 432.3 (M + H)+. d) Ethyl (+)-10,1 l-dihydro-3-[3-(3,4,5,6-tetrahydropyrimid-2-ylamino)-l-propyloxy]-5H-dibenzo[a,d]cycloheptene- 10-acetate 20 A mixture of ethy 1 (±)-10,11 -dihydro-3-[3-(pyrimidin-2-ylamino)-1 -propyloxy ]- 5H-dibenzo[a,d]cycloheptene-10-acetate (0.17g, 0.38 mmol), 10% palladium on charcoal(O.O85g, 0.08 mmol), 4 M HCl in dioxane (0.1 mL, 0.4 mmol) and éthanol (5 mL) wasshaken under H2 (48 psi) for 6 hr, then the catalyst was removed by Filtration throughcelite®. The filtrate was concentrated to give the title compound (0.19 g) as a yellow oil: 25 MS (ES) 436.3 (M + H)+. e) (±)-10,1 l-Dihydro-3-[3-(3,4,5,6-tetrahydropyrimid-2-ylamino)- l-propyloxy]-5H-dibenzo[a,d]cycloheptene-10-acetic acid A solution of ethyl (±)-10,l l-dihydro-3-[3-(3,4,5,6-tetrahydropyrimid-2-ylamino)- 30 l-propyloxy]-5H-dibenzo[a,d]cycloheptene-10-acetate (0.17g, 0.36 mmol), lithium hydroxide monohydrate (0.042g, 1 mmol), THF (3 mL), and water (10 mL) was stirred atroom température for 20 hr, then was concentrated. The residue was dissolved in water,and the solution was brought to pH 4 with 3 N HCl. The resulting solution was kept in therefrigerator ovemight, then the supematant was decanted away from the solid. The solid 35 was dried in vacuum to give the title compound (0.145 g, 91%) as a tan solid: MS (ES)408.3 (M + H)+. Anal. Calcd for C24H29N3O3 · 1.3 HCl: C, 63.37; H, 6.71; N, 9.23.Found: C, 63.67; H, 6.84; N, 9.46. -65 - 011341
Example 9
Préparation of (±)-10,l l-dihvdro-3-F3-(isoquinoline-l-vlamino)-l-propyloxv1-5H- 5 dibenzofa,d1cycloheptene-10-acetic acid a) Ethyl (±)-10,l l-dihydro-3-[3-(l-oxoisoquinoline-l-yIamino)-l-propyloxy]-5H-dibenzo[a,d]cycloheptene-10-acetate s
According to the procedure of Example 6 (a), except substituting l-[(3-hydroxy-l-10 propyl)amino]-isoquinoline N-oxide for the 2-[(3-hydroxy-l-propyl)amino]pyridine-N- oxide, and substituting ethyl (±)-10,l l-dihydro-3-hydroxy-5H-dibenzo[a,d]cycloheptene-10-acetate for the ethyl (R)-10,1 l-dihydro-3-hydroxy-5H-dibenzo[a,d]cycloheptene-10-acetate, the title compound was prepared as a pale yellow oil: MS (ES) m/e 497.2 (M +H)+. 15 b) Ethyl (+)-10,1 l-dihydro-3-[3-(isoquinoline-l-ylamino)-l-propyloxy]-5H-dibenzo[a,d]cycloheptene-10-acetate
According to the procedure of Example 6 (b), except substituting ethyl (±)-10,l 1-dihydro-3-[3-(l-oxoisoquinoline-l-ylamino)-l-propyloxy]-5H-dibenzo[a,d]cycloheptene- 20 10-acetate for the ethyl (R)-10,l l-dihydro-3-[3-(l-oxopyridin-2-ylamino)-l-propyIoxy]-5H-dibenzo[a,d]cycloheptene-10-acetate, the title compound was prepared as a clear oil:MS (ES) m/e 481.3(M + H)+. c) (±)-10,ll-Dihydro-3-[3-(isoquinoline-l-ylamino)-l-propyloxy]-5H- 25 dibenzo[a,d]cycloheptene-10-acetic acid
According to the procedure of Example 6 (c), except substituting ethyl (±)-10,l 1-dihydro-3-[3-(isoquinoline-l-ylamino)-l-propyloxy]-5H-dibenzo[a,d]cycloheptene-10-acetate for the ethyl (R)-10,1 l-dihydro-3-[3-(pyridin-2-ylamino)-l-propyloxy]-5H-dibenzo[a,d]cycloheptene-10-acetate, the title compound was prepared as an amber solid: 30 MS (ES) m/e 453.2(M + H)+. Anal. Calcd for C29H28N2O3 · 1.3 TFA · 0.25 H2O: C,62.71; H, 4.96; N, 4.63. Found: C, 62.45; H, 4.92; N, 4.41. -66- 011 341
Example 10
Préparation of (±)-10,l l-dihvdro-3-i3-i4-(ethvlthio)pvridin-2-vlaminol-l-propyloxvl-5H- dibenzola.dlcycloheptene-10-acetic acid 5 a) Ethyl (±)-10,l l-dihydro-3-[3-[4-(ethylthio)-l-oxopyridin-2-ylamino]-l-propyloxy]-5H-dibenzo[a,d]cycloheptene-10-acetate A solution of ethyl (±)-10,ll-dihydro-3-[3-(4-nitro-l-oxopyridin-2-ylamino)-l-propyloxy]-5H-dibenzo[a,d]cycloheptene-10-acetate (300 mg, 0.61 mmol) and sodium 10 thioethylate (145 mg, 1.22 mmol) in DMF (5 mL) was warmed at 70°C for 3h. The solventwas removed on the rotavap and the residue was purified by silica gel chromatography (2 -6% CH3OH/CH2CI2) to give the title compound (90 mg) as an orange oil: MS (ES) m/e507.3(M + H)+. 15 b) Ethyl (±)-10,1 l-dihydro-3-[3-[4-(ethylthio)pyridin-2-ylamino]-l-propyloxy]-5H-dibenzo[a,d]cycloheptene-10-acetate A mixture of ethyl (±)-10,ll-dihydro-3-[3-[4-(ethylthio)-l-oxopyridin-2-ylamino]-l-propyloxy]-5H-dibenzo[a,d]cycloheptene-10-acetate (60 mg, 0.119 mmol), Fe powder(70 mg), and glacial acetic acid (2 mL) was heated at 100°C for 1.5 h. The mixture was 20 cooled to RT and diluted with H2O and EtOAc, and the pH was adjusted to 7 - 8 with solidNa2CC>3. The layers were separated, and the aqueous layer was extracted with EtOAc.
The combined organic layers were washed with H2O, dried (MgSO4), and concentrated togive the title compound (60 mg) as a yellow oil: MS (ES) m/e 491.3(M + H)+. 25 c) (±)-10,l l-Dihydro-3-[3-[4-(ethylthio)pyridin-2-ylamino]-l-propyloxy]-5H-dibenzo[a,d]cycloheptene- 10-acetic acid
According to the procedure of Example 6 (c), except substituting ethyl (+)-10,11-dihydro-3-[3-[4-(ethylthio)pyridin-2-ylamino]-l-propyloxy]-5H-dibenzo[a,d]cycloheptene-10-acetate for the ethyl (R)-10,1 l-dihydro-3-[3-(pyridin-2-ylamino)-l-propyIoxy]-5H- 30 dibenzo[a,d]cycloheptene-10-acetate, the title compound was prepared as a yellow: ’HNMR (400 MHz, DMSO-d6) δ 7.77 - 7.76 (d, 1 H), 7.17 - 7.15 (d, 1 H), 7.13 - 7.12 (d, 2H), 7.08 - 7.07 (m, 1 H), 6.96 - 6.94(d, 1 H), 6.81 - 6.80 (s, 1 H), 6.68 - 6.67 (d, 1 H), 6.52(s, 1 H), 6.35 - 6.33 (d, 2 H), 6.30 (s, 1 H), 4.20 - 4.16 (d, 1 H), 3.99 - 3.96 (t, 2 H), 3.89 -3.85 (d, 1 H), 3.65 - 3.63 (m, 1 H), 3.36 - 3.32 (m, 2 H), 3.22 - 3.15 (m, 1 H), 2.96 - 2.90 35 . (m, 2 H); 2.85 - 2.78 (m, 1 H), 2.62 - 2.56(m, 2 H), 1.94 - 1.90(m, 2 H), 1.26 - 1.22 (t, 3 H);MS (ES) m/e463.4(M + H)+. -61 - 011341
Exampie i i
Préparation of (±)-10,l l-dihvdro-2-methyl-3-f3-(pvridin-2-vlamino)-l-propyloxv1-5H- dibenzofa,dlcycloheptene-10-acetic acid 5 a) Ethyl (±)-10,l l-dihydro-2-methyl-3-[3-[N-(rerr-butoxycarbonyl)-N-(l-oxopyridin-2-yl)amino]-l-propyloxy]-5H-dibenzo[a,d]cycloheptene-10-acetate
NaH (60% dispersion in minerai oil, 0.14 g, 0.37 mmol) was added to a solution ofethyl (±)-10,l l-dihydro-3-hydroxy-2-methyl-5H-dibenzo[a,d]cycloheptene-10-acetate (100 10 mg, 0.32 mmol) in DMSO (2 mL) under argon, and the reaction was stirred at RT for 0.5hr. A solution of 2-[N-(3-methanesulfonyloxy-l-propyl)-N-(rm-butoxycarbonyl)amino]pyridine-N-oxide (160 mg, 0.4 mmole) in DMSO (1 mL) was thenadded dropwise. The reaction was stirred at RT under argon for 18 hr, then was quenchedwith water (20 mL) and extracted with EtOAc. Drying (MgSO^, concentration, and silica 15 gel chromatography (1% MeOH/CH2Cl2) gave the title compound (85 mg, 42%) as acolorless oil: MS (ES) m/e 561.3 (M + H)+. b) Ethyl (±)-10,11 -dihydro-2-methyl-3-[3-( 1 -oxopyridin-2-ylamino)-1 -propyloxy]-5H-dibenzo[a,d]cycloheptene-10-acetate 20 TFA (0.16 g, 1.4 mmol) was added dropwise to a solution of ethyl (±)-10,l 1- dihydro-2-methyl-3-[3-[N-(reri-butoxycarbonyl)-N-(l-oxopyridin-2-yl)amino]-l-propyloxy]-5H-dibenzo[a,d]cycloheptene-l 0-acetate (80 mg, 0.14 mmole) in dry CH2CI2(3 mL). The reaction was stirred for 5 hr then was concentrated on the rotavap to afford thetitle compound (60 mg, 43%) as a colorless oil: MS (ES) m/e 461.1 (M + H)+. 25 c) Ethyl (±)-10,l l-dihydro-2-methyl-3-[3-(pyridin-2-ylamino)-l-propyloxy]-5H-dibenzo[a,d]cycloheptene-10-acetate
According to the procedure of Example 6 (b), except substituting ethyl (±)-10,l 1-dihydro-2-methyl-3-[3-( 1 -oxopyridin-2-y lamino)-1 -propyloxy]-5H- 30 dibenzo[a,d]cycloheptene-l0-acetate for the ethyl (R)-10,l l-dihydro-3-[3-(l-oxopyridin-2- ylamino)-l-propyloxy]-5H-dibenzo[a,d]cycloheptene-10-acetate, the title compound wasprepared as an off-white solid: MS (ES) m/e 417.3 (M + H)+. d) (±)-10,l l-Dihydro-2-methyl-3-[3-(pyridin-2-ylamino)-l-propyloxy]-5H- 35 dibenzo[a,d]cycloheptene-10-acetic acid
According to the procedure of Example 6 (c), except substituting ethyl (±)-10,11-dihydro-2-methyl-3-(3-(pyridin-2-ylamino)-l-propyloxy]-5H-dibenzo[a,d]cycloheptene- 011341 -68- 10-acetate for the ethyl (R)-10,1 l-dihydro-3-[3-(pyridin-2-ylamino)-l-propyloxy]-5H-dibenzo[a,d]cycloheptene-10-acetate, the title compound was obtained as an off whitesolid: lH NMR (400 MHz, CDC13) δ 7.75 (d, 1 H), 7.65 (t, 1 H), 7.15 (m, 3 H), 7.05 (m, 1H), 6.83 (s, 1 H), 6.7 (d, 1 H), 6.65 (m, 1 H), 6.60 (s, 1 H), 4.25 (d, J = 15.1 Hz, 1 H), 4.05 5 (t, 2 H), 3.80 (m, 1 H), 3.75 (d, J = 15.1 Hz, 1 H), 3.50 (t, 2 H), 3.25 (dd, 1 H), 2.85 (dd, 1 H), 2.68 (dd, 1 H), 2.60 (dd, 1 H), 2.15 (t, 2 H), 2.10 (s, 3 H); MS (ES) m/e 417.3 (M + H)+.
Example 12 10
Préparation of (+)-10,1 l-dihvdro-2-(dimethylamino)methyl-7-fluoro-3-i3-(pyridin-2- vlamino)-l-propyloxy1-5H-dibenzoia,d1cvcloheptene-10-acetic acid a) Ethyl (+)-10,1 l-dihydro-2-(dimethylamino)methyl-7-fluoro-3-[3-(l-oxopyridin-2- 15 ylamino)-1 -propyloxy]-5H-dibenzo[a,d]cycloheptene-10-acetate
According to the procedure of Example 6 (a), except substituting ethyl (±)-10,l 1-dihydro-2-(dimethylamino)methyl-7-fluoro-3-hydroxy-5H-dibenzo[a,d]cycloheptene-10-acetate for the ethyl (R)-10,1 l-dihydro-3-hydroxy-5H-dibenzo[a,d]cycloheptene-10-acetateethyl, the title compound was obtained following silica gel chromatography (gradient: 1:1 20 EtOAc hexanes, then EtOAc, then 20% MeOH/CH2Cl2, then 30% MeOH/CH2Cl2): MS(ES) m/e 522.3 (M + H)+. b) Ethyl (±)-10,11 -dihydro-2-(dimethylamino)methyl-7-fluoro-3-[3-(pyridin-2-ylamino)-1 -propyloxy]-5H-dibenzo[a,d]cycloheptene- 10-acetate 25 According to the procedure of Example 6 (b), except substituting ethyl (±)-10,11 -
dihydro-2-(dimethylamino)methyl-7-fluoro-3-[3-(l-oxopyridin-2-ylamino)-l-propyloxy]-5H-dibenzo[a,d]cycloheptene-10-acetate for the ethyl (R)-10,l l-dihydro-3-[3-(l-oxopyridin-2-ylamino)-l-propyloxy]-5H-dibenzo[a,d]cycloheptene- 10-acetate, the titlecompound was obtained following silica gel chromatography (10% MeOH/CH2Cl2): MS 30 (ES) m/e 506.2 (M + H)+. c) (±)-10,11 -Dihydro-2-(dimethylamino)methyl-7-fluoro-3-[3-(pyridin-2-ylamino)-1 -propyloxÿ]-5H-dibenzo[a,d]cycloheptene- 10-acetic acid
Saponification was conducted according to the procedure of Example 6 (c), except 35 substituting ethyl (±)-10,11 -dihydro-2-(dimethylamino)methyl-7-fluoro-3-(3-(pyridin-2- y lamino)-l-propyloxy]-5H-dibenzo[a,d]cycloheptene- 10-acetate for the ethyl (R)-10,11 -dihydro-3-[3-(pyridin-2-ylamino)-l-propyloxy]-5H-dibenzo[a,d]cycloheptene-10-acetate. -69- 011341
The réaction was acidified with giaeiai HOAc, and the cruae product was desalted bychromatography on XAD-2 resin to afford the title compound as a white solid: MS (ES)m/e 478.3 (M + H)+. Anal. Calcd for C30H36FN3O5 · 1.25 H2O: C, 64.32; H, 6.92; N,7.50. Found: C, 63.87; H, 6.47; N, 7.96.
Example 13
Préparation of (S)-10,l l-dihydro-3-f3-(4-methylpyridin-2-vlamino)-l-propyloxy1-5H- dibenzofa.dlcvcloheptene-10-acetic acid 10 a) Ethyl (S)-10,1 l-dihydro-3-[3-(4-methyl-l-oxopyridin-2-ylamino)-l-propyloxy]-5H-dibenzo[a,d]cycloheptene- 10-acetate A solution of 2-[(3-hydroxy-l-propyl)amino]-4-methylpyridine-N-oxide (1.72 g,9.45 mmole) and diethyl azodicarboxylate ( 1.49 mL, 9.45 mmole) in anhydrous DMF (50 15 mL) was added dropwise over 10 min to a solution of ethyl (S)-10,l l-dihydro-3-hydroxy-5H-dibenzo[a,d]cycloheptene- 10-acetate ( 1.4 g, 4.72 mmole) and triphenylphosphine (2.60g, 9.92 mmole) in anhydrous DMF (50 mL) at RT under argon. After 19 hr, the reactionwas concentrated on the rotavap, and the residue was reconcentrated from xylenes toremove residual DMF. Silica gel chromatography (gradient: 30% EtOAc/hexanes (0.5 L), 20 then EtOAc (1 L), then 5% MeOH/CHCl3) gave the title compound (1.31 g, 60 %) as ayellow oil: MS (ES) m/e 461.3 (M + H)+. b) Ethyl (S)-10,11 -dihydro-3- [3-(4-methylpyridin-2-ylamino)-1 -propy loxy]-5H-dibenzo[a,d]cycloheptene-10-acetate 25 A mixture of ethyl (S)-10,l l-dihydro-3-[3-(4-methyl-l-oxopyridin-2-ylamino)-l- propyloxy]-5H-dibenzo[a,d]cycloheptene-10-acetate (0.86 g, 1.87 mmole), 10%Pd/C (0.86g, 0.81 mmole), cyclohexene (1.89 mL, 18.7 mmole), and isopropanol (20 mL) was heatedat reflux under argon for 19 hr, then the catalyst was renioved by filtration through celite®.Silica gel chromatography (1:9:10 MeOH/CH2Cl2/EtOAc) gave the title compound (0.65 30 g, 78%) as a clear oil: MS (ES) m/e 445.2 (M + H)+. c) (S)-10,11 -Dihydro-3-[3-(4-methylpyridin-2-ylamino)-1 -propyloxy]-5H-dibenzo[a,d]cycloheptene-10-acetic acid
A mixture of ethyl (S)-10,ll-dihydro-3-[3-(4-methylpyridin-2-ylamino)-l-35 propyloxy]-5H-dibenzo[a,d]cycloheptene- 10-acetate (2.08 g, 4.69 mmole) and 1.0 N NaOH (7.0 mL, 7.0 mmole) in absolute EtOH (45 mL) was warmed in an oil bath set at 45°C.After 18 hr, the reaction was concentrated on the rotavap and the pH was adjusted to 7 with 011341 -ΊΟ- 1.0 N HCl. The solid precipitate was collected and washed with H2O. Drying ovemightafforded the title compound (1.61 g, 82%) as a nearly colorless solid: MS (ES) m/e 417.4(M + H)+. Anal. Calcd for C26H28N2O3 · 1.0 H2O: C, 71.87; H, 6.96; N, 6.45. Found: C, 71.63; H, 6.96; N, 6.30. 5
Example 14
Préparation of (S)-10,l l-dihvdro-3-f3-f4-(2-propyloxv)pyridin-2-vlamino1-l-propyloxv1- 5H-dibenzoia,dlcycloheptene- 10-acetic acid 10 a) Isopropyl (S)-10,ll-dihydro-3-[3-[4-(2-propyloxy)-l-oxopyridin-2-ylamino]-l-propyloxy]-5H-dibenzo[a,d]cycloheptene-l 0-acetate A mixture of ethyl (S)-10,1 l-dihydro-3-[3-(4-nitro-l-oxopyridin-2-y lamino)-1-propyloxy]-5H-dibenzo[a,d]cycloheptene-10-acetate (0.2 g, 0.4 mmole) and sodium 15 isopropoxide (0.067 g, 0.8 mmole) in isopropanol (5 mL) was heated at 80°C for 3.5 hr, then more sodium isopropoxide (0.05 g, 0.6 mmole) was added, and the reaction was stirredat RT ovemight. Concentration and silica gel chromatography (gradient: 5% - 15%MeOH/CH2Cl2) gave the title compound (0.106 g, 52%) as a light brown oil: MS (ES)519.3 (M + H)+. 20 b) Isopropyl (S)-10,l l-dihydro-3-[3-[4-(2-propyloxy)pyridin-2-ylamino]-l-propyloxy]-5H-dibenzo[a,d]cycloheptene-10-acetate
According to the procedure of Example 13 (b), except substituting isopropyl (S)-10,1 l-dihydro-3-[3-[4-(2-propyloxy)-l-oxopyridin-2-y lamino]-l-propyloxy]-5H- 25 dibenzo[a,d]cycloheptene-l0-acetate for the ethyl (S)-10,1 l-dihydro-3-[3-(4-methyl-l- oxopyridin-2-ylamino)-l-propyloxy]-5H-dibenzo[a,d]cycloheptene-10-acetate, the titlecompound was obtained as slightly yellow oil following silica gel chromatography (5%MeOH/CH2Cl2): MS (ES) 503.4 (M + H)+. 30 c) (S)-10,1 l-Dihydro-3-[3-[4-(2-propyloxy)pyridin-2-ylamino]-l-propyloxy]-5H-dibenzo[a,d]cycloheptene- 10-acetic acid
According to the procedure of Example 13 (c), except substituting isopropyl (S)-10,1 l-dihydro-3-[3-[4-(2-propy loxy)pyridin-2-y lamino]-1 -propyloxy]-5H-dibenzo[a,d]cycloheptene-10-acetate for the ethyl (S)-10,l l-dihydro-3-[3-(4- 35 methylpyridin-2-ylamino)-l-propyloxy]-5H-dibenzo[a,d]cycloheptene-l0-acetate, the titlecompound was obtained as white powder: MS (ES) 461.3 (M + H)+. Anal. Calcd forC28H32N2°4 · °·96 HC!: C, 67.86; H, 6.70; N, 5.65. Found: C, 68.26; H, 6.86; N, 5.25. -71 - 011341
Example 15
Préparation of (S)-10.1 l-dihvdro-3-i3-(4-chloropyridin-2-vlamino)-l-propvloxvl-5H- 5 dibenzora.dlcvcloheptene-10-acetic acid a) Ethyl (S)-10,l l-dihydro-3-[3-(4-chloro-l-oxopyridin-2-ylamino)-l-propyloxy]-5H-dibenzo[a,d]cycloheptene-10-acetate A solution of ethyl (S)-10,l l-dihydro-3-[3-(4-nitro-l-oxopyridin-2-ylamino)-l-10 propyloxy]-5H-dibenzo[a,d]cycloheptene-10-acetate (0.47 g, 0.96 mmole) in acetyl chloride (7 mL, 98 mmole) was heated at reflux for 1 hr. The reaction mixture was pouredonto ice (50 g), and the pH was adjusted to 8.0 using saturated NaHCO3 (caution: bubblesviolently!). The mixture was extracted with CH2CI2 (2 x 100 mL), and the combinedorganic layers were washed sequentially with H2O (50 mL) and brine (50 mL). Drying 15 (MgSC>4) and concentration gave the title compound: MS (ES) 481.2 (M + H)+. b) Ethyl (S)-10,1 l-dihydro-3-[3-(4-chloropyridin-2-ylamino)-l-propyloxy]-5H-dibenzo[a,d]cycloheptene-10-acetate A mixture of ethyl (S)-10,1 l-dihydro-3-[3-(4-chloro-l-oxopyridin-2-ylamino)-l-20 propyloxy]-5H-dibenzo[a,d]cycloheptene-l0-acetate (0.13 g, 0.27 mmole) and 2.0 M PCI3 in CH2CI2 (8 mL, 16 mmole) was heated at reflux for 22 hr. The reaction mixture wascooled and poured onto ice (200 g), and the pH was adjusted to 12 using 40% NaOH.CH2CI2 (2 x 100 mL) extraction, drying (MgSO^, concentration, and silica gelchromatography (4% MeOH/CH2Cl2) gave the title compound (93 mg, 74%) as light 25 yellow oil: MS (ES) 465.3 (M + H)+. c) (S)-10,1 l-Dihydro-3-[3-(4-chloropyridin-2-ylamino)-l-propyloxy]-5H-dibenzo[a,d]cycloheptene-10-acetic acid
According to the procedure of Example 13 (c), except substituting ethyl (S)-10,l 1-30 dihydro-3-[3-(4-chloropyridin-2-ylamino)-l-propyloxy]-5H-dibenzo[a,d]cycloheptene-10- acetate for the ethyl (S)-10,l l-dihydro-3-[3-(4-methylpyridin-2-ylamino)-l-propyloxy]-5H-dibenzo[a,d]cycloheptene-10-acetate, the title compound was obtained as off-whitepowder: MS (ES) 437.2 (M + H)+. Anal. Calcd for C25H25N2O3 · 1.0 HCl: C, 63.43; H,5.54; N, 5.92. Found: C, 63.11; H, 5.82; N, 5.62. 35 -72- 011341
Example 16
Préparation of (S)-10.1 l-dihydro-3-[3-i4-(dimethylamino)pyridin-2-ylaminol-l- propyloxvl-5H-dibenzo[a,dlcvcloheptene-10-acetic acid 5 a) Ethyl (S)-10,1 l-dihydro-3-(3-(4-chloro- l-oxopyridin-2-ylamino)-l-propyloxy]-5H-dibenzo[a,d]cycloheptene-10-acetate A solution of ethyl (±)-10,l l-dihydro-3-[3-(4-nitro-l-oxopyridin-2-ylamtno)-l-propyloxy]-5H-dibenzo[a,d]cycloheptene-10-acetate (0.47 g, 0.96 mmole) in acetyl 10 chloride (7 mL, 98 mmole) was heated at reflux for 1 hr. The reaction mixture was pouredonto ice (50 g), and the pH was adjusted to 8.0 using saturated NaHCOj (caution: bubblesviolently!). The mixture was extracted with CH2CI2 (2 x 100 mL), and the combinedorganic layers were washed sequentially with H2O (50 mL) and brine (50 mL). Drying(MgSOzj.) and concentration gave crude title compound which was carried forward without 15 further purification. MS (ES) 481.3 (M + H)+. b) Ethyl (S)-10,11 -dihydro-3-[3-[4-(dimethylamino)-1 -oxopyridin-2-ylamino]-1 -propy loxy]-5H-dibenzo[a,d]cycloheptene-10-acetate A mixture of ethyl (S)-10,1 l-dihydro-3-[3-(4-chloro-l-oxopyridin-2-y lamino)-1-
20 propyloxy]-5H-dibenzo[a,d]cycloheptene-10-acetate (0.96 mmole) and 2.0 M dimethylamine in MeOH (3 mL, 6 mmole) was refluxed for 16 hr. Concentration and silicagel chromatography (7% MeOH/CH2Cl2) gave the title compound (0.049 g, 10 %) as alight brown powder. MS (ES) 490.3 (M + H)+. Unchanged ethyl (S)-10,1 l-dihydro-3-[3-(4-chloro-l-oxopyridin-2-ylamino)-l-propyloxy]-5H-dibenzo[a,d]cycloheptene-10-acetate 25 was also recovered from the chromatographie purification. c) Ethyl (S)-10,l l-dihydro-3-[3-[4-(dimethylamino)pyridin-2-ylamino]-l-propyloxy]-5H-dibenzo[a,d]cycloheptene-10-acetate
According to the procedure of Example 13 (b), except substituting ethyl (S)-10,11- 30 dihydro-3-[3-[4-(dimethylamino)-1 -oxopyridin-2-y lamino]-1 -propyloxy]-5H- dibenzo[a,d]cycloheptene-l 0-acetate for the ethyl (S)-10,ll-dihydro-3-[3-(4-methyl-l-oxopyridin-2-ylamino)-l-propyloxy]-5H-dibenzo[a,d}cycloheptene-10-acetate, the titlecompound was obtained as white powder following silica gel chromatography (8%MeOH/CH2Cl2): MS (ES) 474.3 (M + H)+. 35 011341 -n- d) (S)-!O,! !-Dihydro-3-[3-[4-(dimethylamino)pyridin-2-ylaminoj-l-propyioxy]-5H-dibenzo[a,d]cycloheptene-10-acetic acid
According to the procedure of Example 13 (c), except substituting ethyl (S)-10,l 1-dihydro-3-[3-[4-(dimethylamino)pyridin-2-ylamino]-l-propyloxy]-5H- 5 dibenzo[a,d]cycloheptene-10-acetate for the ethyl (S)-10,1 l-dihydro-3-[3-(4- methylpyridin-2-ylamino)-1 -propyloxy]-5H-dibenzo[a,d]cycloheptene-10-acetate, the titlecompound was obtained as white powder: MS (ES) 446.2 (M + H)+. Anal. Calcd forC27H31N3O3 · 0.5 H2O · 1.0 HCl: C, 66.04; H, 6.77; N, 8.56. Found: C, 65.96; H, 6.60;N, 8.26. 10
Example 17
Préparation of (S)-10.1 l-dihvdro-3-i3-(4-ethoxvpyridin-2-ylamino)-l-propyloxvl-5H- dibenzofa,dlcycloheptene-l 0-acetic acid 15 a) Ethyl (S)-10,l l-dihydro-3-[3-(4-ethoxy-l-oxopyridin-2-ylamino)-l-propyloxy]-5H-dibenzo[a,d]cycloheptene-10-acetate
According to the procedure of Example 2 (a), except substituting ethyl (S)-10,11-dihydro-3-[3-(4-nitro-1 -oxopyridin-2-y Iamino)-1 -propyloxy]-5H- 20 dibenzo[a,d]cycloheptene-l0-acetate (496.9 mg, 1.01 mmol) for the ethyl (±)-10,l 1-dihydro-3- [3-(4-nitro-1 -oxopy ridin-2-y Iamino)-1 -propyloxy]-5H-dibenzo[a,d]cycloheptene-l 0-acetate, and using 0.53 M NaOEt (4.0 mL, 2.12 mmol) andabsolute éthanol (10 mL) in the displacement reaction, the title compound (456.2 mg, 92%)was prepared: MS (ES) m/e 491 (M + H)+. 25 b) Ethyl (S)-10,l l-dihydro-3-[3-(4-ethoxypyridin-2-ylamino)-l-propyloxy]-5H-dibenzo[a,d]cycloheptene-10-acetate
According to the procedure of Example 2 (b), except substituting ethyl (S)-10,11-dihydro-3-[3-(4-ethoxy-l-oxopyridin-2-ylamino)-l-propyloxy]-5H- 30 dibenzo[a,d]cycloheptene-l 0-acetate (456.2 mg, 0.93 mmole) for the ethyl (±)-10,l 1-dihydro-3-[3-(4-ethoxy-l-oxopyridin-2-ylamino)-l-propyloxy]-5H-dibenzo[a,d]cycloheptene-l 0-acetate, the title compound (475.2 mg, quantitative) wasprepared: MS (ES) m/e 475 (M + H)+. 011341 - 74- c) (S)-10,11 -Dihydro-3-[3-(4-ethoxypyridin-2-ylamino)-1 -propy loxy ]-5H-dibenzo[a,d]cycloheptene-10-acetic acid 1.0 N NaOH (2.0 mL, 2.0 mmole) was added to a solution of ethyl (S)-10,11 -dihydro-3-[3-(4-ethoxypyridin-2-ylamino)-l-propyloxy]-5H-dibenzo[a,d]cycloheptene-10- 5 acetate (475.2 mg, 1.0 mmol) in absolute éthanol (10 mL), and the solution was warmed at50°C in an oil bath. After 20 hr, the reaction was concentrated and the aqueous residue wascooled to 0°C in an ice bath. 1.0 N aqueous HCl (2.0 mL, 2.0 mmole) was slowly addedwith stirring. An opaque solid residue precipitated and was collected on a sintered glassfunnel. Drying in a vacuum desiccator ovemight gave the title compound (452.6 mg, 83%): 10 MS (ES) m/e 447 (M + H)+. Anal. Calcd for C27H3oN204 · 0.20 H2O 1.75 HCl: C,63.10; H, 6.30; N, 5.45. Found: C, 63.10; H, 5.98; N, 5.38.
Example 18 15 Préparation of (±)-10,l l-dihydro-7-f1uoro-3-f3-(pyridin-2-ylamino)-l-propyloxyl-5H- dibenzofa.dlcycloheptene-10-acetic acid a) Ethyl (±)-10,l l-dihydro-7-fluoro-3-[3-(l-oxopyridin-2-ylamino)-l-propy loxy ]-5H-dibenzo[a,d]cycloheptene-10-acetate 20 According to the procedure of Example 6 (a), except substituting ethyl (±)-10,11- dihydro-7-fluoro-3-hydroxy-5H-dibenzo[a,d]cycloheptene-10-acetate for the ethyl (R)-10,U-dihydro-3-hydroxy-5H-dibenzo[a,d]cycloheptene-10-acetate, the title compound wasobtained as a colorless oil following silica gel chromatography (gradient: 1:1EtOAc/hexanes, then EtOAc, then 4% MeOH/CH2Cl2): MS (ES) m/e 465.3 (M + H)+. 25 b) Ethyl (±)-10,1 l-dihydro-7-fluoro-3-[3-(pyridin-2-ylamino)-l-propyloxy]-5H-dibenzo[a,d]cy cloheptene-10-acetate
According to the procedure of Example 6 (b), except substituting ethyl (±)-10,11-dihydro-7-fluoro-3-[3-(l-oxopyridin-2-ylamino)-l-propy loxy]-5H- 30 dibenzo(a,d]cycloheptene-10-acetate for the ethyl (R)-10,ll-dihydro-3-[3-(l-oxopyridin-2-ylamino)-l-propyloxy]-5H-dibenzo[a,d]cycloheptene-l 0-acetate, the title compound wasobtained: MS (ES) m/e 449.2 (M + H)+. c) (±)-10,l l-Dihydro-7-fluoro-3-[3-(pyridin-2-ylamino)-l-propyloxy]-5H- 35 dibenzo[a,d]cycloheptene-10-acetic acid
According to the procedure of Example 6 (c), except substituting ethyl (±)-10,l 1-dihydro-7-fluoro-3-[3-(pyridin-2-ylamino)-l-propyloxy]-5H-dibenzo[a,d]cycloheptene-10- -75- 011341 acetate for the ethyl (R)-10,1 l-dihydro-3-[3-(pyridin-2-y!amir.o)-l-propyloxyj-5K-dibenzo[a,d]cycloheptene-10-acetate, the title compound was obtained: MS (ES) m/e 421.1(M + H)+. Anal. Calcd for C25H25FN2O3 · 0.5 H2O: C, 69.99; H, 6.10; N, 6.52. Found:C, 69.86; H, 5.90; N, 6.35. 5
Example 19
Préparation of (t)-10,l l-dihvdro-6-methvl-3-i3-(pvridin-2-ylamino)-l-propyloxvl-5H- dibenzofa,d1cycloheptene-10-acetic acid 10 a) Ethyl (±)-10,l l-dihydro-6-methy 1-3-(3-( 1-oxopyridin-2-ylamino)-l-propyloxy]-5H-dibenzo[a,d]cycloheptene-10-acetate
According to the procedure of Example 6 (a), except substituting ethyl(±)-10,l 1-dihydro-3-hydroxy-6-methyl-5H-dibenzo[a,d]cycloheptene-10-acetate for the ethyl (R)- 15 10,1 l-dihydro-3-hydroxy-5H-dibenzo[a,d]cycloheptene-10-acetate, the title compound was obtained as a colorless oil following silica gel chromatography (gradient: 1:1EtOAcZhexanes, then EtOAc, then 4% MeOH/CH2Cl2): MS (ES) m/e 461.3 (M + H)+. b) Ethyl (±)-10,l l-dihydro-6-methyl-3-[3-(pyridin-2-ylamino)-l-propyloxy)-5H- 20 dibenzo[a,d]cycloheptene- 10-acetate
According to the procedure of Example 6 (b), except substituting ethyl (±)-10,11 -dihydro-6-methyl-3-[3-(l-oxopyridin-2-ylamino)-l-propyloxy]-5H-dibenzo[a,d]cycloheptene-10-acetate for the ethyl (R)-10,1 l-dihydro-3-[3-(l-oxopyridin-2-ylamino)-l-propyloxy]-5H-dibenzo[a,d]cycloheptene-10-acetate, the title compound was 25 obtained following silica gel chromatography (1% MeOH/CH2Cl2): MS (ES) m/e 445.3(M + H)+. c) (±)-10,11 -Dihydro-6-methyl-3-[3-(pyridin-2-ylamino)-l -propyloxy]-5H-dibenzo[a,d]cycloheptene-10-acetic acid 30 According to the procedure of Example 6 (c), except substituting ethyl (±)-10,11 - dihydro-6-methyl-3-[3-(pyridin-2-ylamino)-l-propyloxy]-5H-dibenzo[a,d]cycloheptene-10-acetate for the ethyl (R)-10,1 l-dihydro-3-[3-(pyridin-2-ylamino)- l-propyloxy]-5H-dibenzo[a,d]cycloheptene-10-acetate, the title compound was obtained as a white solid: MS (ES) m/e 417.3 (M + H)+. Anal. Calcd for C26H28N2O3 · 1.25 H2O: C, 71.13; H, 35 7.02; N, 6.38. Found: C, 71.33; H, 6.67; N, 6.01. -76- 011341
Example 20
Préparation of (S)-10,l l-dihvdro-3-f3-(4-aminopvridin-2-vlamino)-l-propyloxyl-5H- 5 dibenzofa,dlcycloheptene-10-acetic acid a) Ethyl (S)-10,1 l-dihydro-3-[3-(4-nitro-l-oxopyridin-2-ylamino)-l-propyloxy]-5H-dibenzo[a,d]cycloheptene-10-acetate A solution of diisopropyl azodicarboxylate (1.7 mL, 8 mmole) in THF (10 mL) was 10 added dropwise to a solution of ethyl (S)-10,1 l-dihydro-3-hydroxy-5H- dibenzo[a,d]cycloheptene-10-acetate (426.5 mg, 1.5 mmole), 2-[(3-hydroxy-l-propyl)amino]-4-nitropyridine-N-oxide (1.7 g, 8 mmole), and triphenylphosphine (2.5 g, 8mmole) in anhydrous DMF (20 mL) at 0 °C under argon. The yellow solution was kept at 0°C for 10 min, then was warmed to RT. After 23 hr, the reaction was concentrated. Silica 15 gel chromatography (gradient: 30% - 100% EtOAc/hexanes) gave the title compound (2.7g, 81%) as an orange foam: MS (ES) m/e 491.8 (M + H)+. b) Ethyl (S)-10,l l-dihydro-3-[3-(4-aminopyridin-2-ylamino)-l-propyloxy]-5H-dibenzo[a,d]cyc loheptene-10- acetate 20 A mixture of ethyl (S)-10,l l-dihydro-3-[3-(4-nitro-l-oxopyridin-2-ylamino)-l- propyloxy]-5H-dibenzo[a,d]cycloheptene-10-acetate (2.7 g, 6 mmole), cyclohexene (6 mL,60 mmole), 10% Pd/C (1.2 g, 1.10 mmole), and isopropanol (30 mL) was heated at refluxunder argon for 20.5 hr, then was hot-filtered through celite®. The fïlter pad was washedwith hot EtOAc, and the combined filtrâtes were concentrated. The residue was 25 chromatographed on silica gel (5% MeOH/CHCl3) to afford the title compound (2.4 g, 98%) as a colorless foam: MS (ES) m/e 445.9 (M + H)+. c) (S)-10,1 l-Dihydro-3-[3-(4-aminopyridin-2-ylamino)-1 -propyloxy]-5H-dibenzo[a,d]cycloheptene-10-acetic acid 30 A mixture of ethyl (S)-10,1 l-dihydro-3-[3-(4-aminopyridin-2-ylamino)-l- propyloxy]-5H-dibenzo[a,d]cycloheptene-10-acetate (2.4 g, 5 mmole), LiOH · H2O (0.3 g, 7 mmole), THF (30 mL), and H2O (10 mL) was stirred at RT for 48 hr, then wasconcentrated. The residue was diluted with H2O and extracted with Et2O. The Et2Ûlayers were discarded. The aqueous layer was stirred with gentle warming under vacuum to 35 remove residual organic solvents, then was filtered. The resulting aqueous solution wasstirred at RT while the pH was slowly and carefully adjusted to 5.5 - 6.0 with 1.0 N HCI.The mixture was stirred for 0.5 hr, then the solid was collected by suction filtration and 011341 -77- washed with plenty of hBO, Drying in high vacuum at 60 °C gave ihe title compound ( î .0g, 42%) as a glassy solid: MS (ES) m/e 417.7 (M + H)+. Anal. Calcd for C25H27N3O3 ·1.4 HCI (468.554): C, 64.08; H, 6.11; N, 8.97. Found: C, 64.16; H, 6.20; N, 8.71. 5 Example 21
Préparation of (±)-10,l l-dihvdro-3-f3-(4-methylpyridin-2-vlamino)-l- propy loxvldibenzof b.floxepine-10-acetic acid 10 a) Ethyl (±)-10,l l-dihydro-3-[3-(4-methyl-l-oxopyridin-2-ylamino)-l-propyloxy]dibenzo[b,f]oxepine-10-acetate A mixture of ethyl (±)-10,l l-dihydro-3-hydroxydibenzo[b,f]oxepine-l 0-acetate(257 mg, 0.86 mmole), 2-[(3-bromo-l-propyl)amino]-4-methylpyridine-N-oxidehydrobromide (308 mg, 0.94 mmole), NaOH pellets (110 mg, 2.75 mmole), and CH3CN (4 15 mL) was stirred at RT under argon ovemight. The mixture was filtered and the solids werewashed with CH3CN. The filtrate was concentrated, and the residue was flashchromatographed on silica gel (1 - 2.5% CH3OH/CH2CI2) to afford the title compound(190 mg, 48%) as a white foam: MS (ES) m/e 462.6 (M + H)+. 20 b) Ethyl (±)-10,1 l-dihydro-3-[3-(4-methylpyridin-2-ylamino)-l-propyloxy]dibenzo[b,f]oxepine-10-acetate A mixture of ethyl (±)-10,l l-dihydro-3-[3-(4-methyl-l-oxopyridin-2-ylamino)-l-propyloxy]dibenzo[b,f]oxepine-10-acetate (183 mg, 0.4 mmole), 10% Pd/C (85 mg, 0.08mmole), cyclohexene (810 mg, 8 mmole), and isopropanol (4 mL) was heated at reflux 25 ovemight. The catalyst was removed by filtration through celite®, and the filter cake waswashed with ether. The filtrate was concentrated to afford the title compound (122 mg,68%) as a clear oil: MS (ES) m/e 446.9 (M + H)+. c) (±)-10,ll-Dihydro-3-[3-(4-methylpyridin-2-ylamino)-i-propyloxy]dibenzo[b,f]oxepine- 30 10-acetic acid A mixture of ethyl (±)-10,ll-dihydro-3-[3-(4-methylpyridin-2-ylamino)-l-propyloxy]dibenzo[b,f]oxepine-l 0-acetate (119 mg, 0.27 mmole) and 0.991 N NaOH(0.545 mL, 0.54 mmole) in absolute EtOH (2 mL) was warmed in an oil bath set at 45 °C.After 20 hr, the reaction was concentrated on the rotavap, and the residue was dissolved in 35 H2O (1.5 mL). The solution was filtered to remove insoluble material, and the filtrate wascarefully neutralized by dropwise addition of 1.0 N HCl (0.54 mL, 0.54 mmole). Theprecipitate was collected and dried in high vacuum to afford the title compound (68 mg, 011341 -78- 58%) as a white solid: MS (ES) m/e 418.9 (M + H)+. Anal. Calcd for C25H26N2O4 · 0.45HCl: C, 69.05; H, 6.13; N, 6.44. Found: C, 69.25; H, 6.27; N, 6.16.
Example 22 5
Préparation of (±)-10,l l-dihydro-3-f2-f6-(methylamino)pyridin-2-yll-l- ethoxvldibenzoib.floxepine-10-acetic acid a) Ethyl (±)-10,l l-dihydro-3-[2-[6-[N-tert-butoxycarbonyl)-N-methylamino]pyridin-2-yl]- 10 l-ethoxy]dibenzo[b,f]oxepine-10-acetate A solution of 6-[N-(tert-butoxycarbonyl)-N-methylamino]-2-pyridyIethanol (397 mg, 1.58 mmole) and diisopropyl azodicarboxylate (0.31 mL, 1.58 mmole) in anhydrousCH2CI2 (8 mL) was added dropwise over 10 min to a solution of ethyl (±)-10,l 1-dihydro-3-hydroxydibenzo[b,f]oxepine-10-acetate (186 mg, 0.63 mmole) and triphenylphosphine 15 (413 mg, 1.58 mmole) in anhydrous CH2CI2 (3.2 mL) at RT under argon. After 22 hr, the reaction was concentrated on the rotavap, and the residue was flash chromatographed onsilica gel (2 - 13% EtOAc/hexanes) to give the title compound (146 mg, 44%) as a clear oil:MS (ES) m/e 533.0 (M+ H)+. 20 b) Ethyl (+)-10,1 l-dihydro-3-[2-[6-(methylamino)pyridin-2-yl]-l-ethoxy]dibenzo[b,f]oxepine- 10-acetic acid 4 N HCl in dioxane (1.3 mL, 5.2 mmole) was added dropwise to a solution of ethyl(±)-10,l l-dihydro-3-[2-[6-[N-tert-butoxycarbonyl)-N-methylamino]pyridin-2-yl]-l-ethoxy]dibenzo[b,f]oxepine-10-acetate (140 mg, 0.26 mmole) in CH2CI2 (1.3 mL) After 25 12 hr, the mixture was concentrated, and the residue was triturated with ether to afford the title compound as a white solid: MS (ES) m/e 432.9 (M + H)+. c) (±)-10,1 l-Dihydro-3-[2-[6-(methylamino)pyridin-2-yl]- l-ethoxy]dibenzo[b,f]oxepine-10-acetic acid 30 A mixture of ethyl (±)-10,l l-dihydro-3-[2-[6-(methylamino)pyridin-2-yl]-l- ethoxy]dibenzo[b,f]oxepine-10-acetic acid (0.26 mmole) and 0.991 N NaOH (0.525 mL,0.52 mmole) in absolute EtOH (2 mL) was warmed iri an oil bath set at 50 °C. After 20 hr,the réaction was concentrated on the rotavap and the residue was dissolved in H2O (1.5mL). The solution was filtered to remove insoluble material, and the filtrate was carefully 35 neutralized by dropwise addition of 1.0 N HCl. The precipitate was collected and dried inhigh vacuum to afford the title compound (72 mg, 30% for 2 steps) as an off-white solid: 011341 - 79- MS (ES) m/e 405.0 (M + H)+. Anal. Calcd for C24H24N2O4 i .25 HCl · 0.25 H2O: C,63.42; H, 5.71; N, 6.16. Found: C, 63.35; H, 5.9; N, 6.16.
Example 23 5
Préparation of (5)-10,1 l-dihvdro-3-i3-(2-aminopvridin-4-yl)-l-propyloxy1-5//- dibenzora.Jlcycloheptene-10-acetic acid a) 3-(2-Aminopyridin-4-yl)propan-l-ol 10 A suspension of 3-(2-aminopyridin-4-yl)propanoic acid hydrochloride (0.73 g, 3.60 mmol, prepared according to WO94/14776 in THF (10 mL) was added over 45 min tolithium aluminum hydride (12 mL, 12 mmol, IM in THF) at 0 °C. The ice bath wasremoved and the reaction was allowed to stir at RT for 4.5 h. The reaction was cooled to 0°C, diluted with toluene (22 mL) and quenched by the sequential addition of H:O (0.86 mL) 15 and NaF (1.54 g). The resulting suspension was stirred at 0 °C for 45 min. The reactionmixture was filtered and the precipitate was washed with additional 10% MeOH in CHC13.The combined filtrâtes were concentrated under reduced pressure. Flash chromatography(10% MeOH/CHCl3, silica gel) gave 0.25 g of the desired material as a clear oil: MS(ES+)m/z 152.7 [M+H]\ 20 b) Ethyl (5)-10,1 l-dihydro-3-[3-(2-aminopyridin-4-yl)-l-propyloxy]-5//-dibenzo[a, J]cycloheptene-10-acetate A solution of Example l(a) (0.23 g, 1.51 mmol) and di-isopropylazadicarboxylate(0.29 mL, 1.50 mmol) in CH2C12 (7.5 mL) was added dropwise to a solution of
25 triphenylphosphine (0.39 g, 1.50 mmol) and ethyl 2-[(10S)-3-hydroxy-10,1 l-dihydro-5H-dibenzo[a,J]cyclohepten-10-yl]acetate (0.30 g, 1.00 mmol) in CH2C1, (5 mL) at 0 °C. Theice bath was removed and the reaction was allowed to warm to RT. After 18 h, the solventwas removed under reduced pressure. Flash chromatogrpàhy (50% EtOAc/hexanes to100% EtOAc, silica gel) gave 0.32 g of material that contained the desired product. A 30 second purification by flash chromatography (75% to 90% EtOAc/hexanes, silica gel) gave0.23 g of the desired material: MS(ES+) m/z 430.9 [M+H]\ c) (S)-10,ll-dihydro-3-[3-(2-aminopyridin-4-yl)-l-propyloxy]-5H-dibenzo[a,d]cycloheptene-10-acetic acid
35 The compound of Example 1 (b) (0.22 g, 0.50 mmol) was dissolved in IN NaOH (0.77 mL, 0.77 mmol), EtOH (3 mL) and THF (3 mL). After heating the reaction at 50 °Cfor 18 h, the solvent was removed under reduced pressure. The residue was dissolved in 011341 -80- Η,Ο (4 mL) and filtered. The filtrate was acidified with 30% TFA in H,O and the resultingprecipitate was collected. Préparative HPLC (Hamilton PRP-1®, 3% CH,CN/H,0 - 0.1 %TFA) gave 10 mg of the desired material as a hygroscopic solid: MS(ES+) m/z 402.6[Μ+Η]\ 5
Example 24
Parentéral Dosage Unit Composition 10 A préparation which contains 20 mg of the compound of Example l as a stérile dry powder is prepared as follows: 20 mg of the compound is dissolved in 15 mL of distilledwater. The solution is filtered under stérile conditions into a 25 mL multi-dose ampouleand lyophilized. The powder is reconstituted by addition of 20 mL of 5% dextrose in water(D5W) for intravenous or intramuscular injection. The dosage is thereby determined by the 15 injection volume. Subséquent dilution may be made by addition of a metered volume ofthis dosage unit to another volume of D5W for injection, or a metered dose may be addedto another mechanism for dispensing the drug, as in a bottle or bag for IV drip infusion orother injection-infusion System. 20 Example 25
Oral Dosage Unit Composition A capsule for oral administration is prepared by mixing and milling 50 mg of the 25 compound of Example 1 with 75 mg of lactose and 5 mg of magnésium stéarate. Theresulting powder is screened and filled into a hard gelatin capsule.
Example 26 30 Oral Dosage Unit Composition A tablet for oral administration is prepared by~ mixing and granulating 20 mg ofsucrose, 150 mg of calcium sulfate dihydrate and 50 mg of the compound of Example 1with a 10% gelatin solution. The wet granules are screened, dried, mixed with 10 mg 35 starch, 5 mg talc and 3 mg stearic acid; and compressed into a tablet. -81 - 011341
The above description fully discloses how to make and use the présent invention.However, the présent invention is not limited to the particular embodiments describedhereinabove, but includes ail modifications thereof within the scope of the followingdaims. The various references to journals, patents and other publications which are citedherein comprises the State of the art and are incorporated herein by référencé as thoughfully set forth.
Claims (33)
- -82- 011341 What is claimed is:
- 3. A compound according to claim 1 in which Y is, wherein each R" is H or Cj^alkyl.
- 4. A compound according to claim 1 in which Y is 10 and s is 1.wherein each R" independently is H or CMalkyl, 5. A compound according to claim 1 in which Y is 15, wherein G is S and each R" independently is H or Cj^alkyl.
- 6. A compound according to claim 1 in which Y is 20
- 7. A compound according to claim 1 which is: (±)-10,11 -Dihydro-3-[2-(6-aminopyridin-2-yl)-1 -ethoxy ]-5H-25 dibenzo[a,d]cycloheptene-10-acetic acid; 011341 -84- (±)-10,11 -Dihydro-3-[4-(pyridin-2-y lamino)-1 -buty 1]-5H-dibenzo[a,d]cycloheptene-10-acetic acid; (±)-10,11 -Dihydro-3-[3-(4-ethoxypyridin-2-y lamino)-1 -propy loxy ]-5H-dibenzo[a,d]cycloheptene-10-acetic acid; 5 (S)-10,1 l-Dihydro-3-[3-(pyridin-2-ylamino)-l-propyloxy]-5H- dibenzo[a,d]cycloheptene-10-acetic acid; (R) -10,1 l-Dihydro-3-[3-(pyridin-2-ylamino)-l-propyloxy]-5H-dibenzo[a,d]cycloheptene- 10-acetic acid; (±)-10,l l-Dihydro-3-[3-(3,4,5,6-tetrahydropyrimidin-2-ylamino)-l-propyloxy]-5H-10 dibenzo[a,d]cycloheptene-10-acetic acid; (±)-10,1 l-Dihydro-3-[2-[2-(ethylamino)thiazol-4-yl]-l-ethoxy]-5H-dibenzo[a,d]cycloheptene- 10-acetic acid; (±)-10,11 -Dihydro-3-[3-(isoquinoline-l -ylamino)-1 -propy loxy]-5H-dibenzo[a,d]cycloheptene- 10-acetic acid; 15 (±)-10,11 -Dihydro-7-fluoro-3-[3-(pyridin-2-y lamino)-1 -propyloxy]-5H- dibenzo[a,d]cycloheptene- 10-acetic acid; (S) -10,1 l-Dihydro-3-[3-(4-methy lpyridin-2-y lamino)-1 -propyloxy]-5H-dibenzo[a,d]cycloheptene-10-acetic acid; (S)-10,11 -Dihydro-3-[3-(4-ethoxypyridin-2-ylamino)-1 -propyloxy]-5H-20 dibenzo[a,d]cycloheptene-10-acetic acid; (±)-10,1 l-Dihydro-6-methyl-3-[3-(pyridin-2-ylamino)- l-propyloxy]-5H-dibenzo[a,d]cycloheptene- 10-acetic acid; (+)-10,1 l-Dihydro-2-(dimethylamino)methyl-7-fluoro-3-[3-(pyridin-2-ylamino)-l-propyloxy]-5H-dibenzo[a,d]cycloheptene-10-acetic acid; 25 (S)-10,1 l-Dihydro-3-[3-[4-(2-propyloxy)pyridin-2-ylamino]-l-propyloxy]-5H- dibenzo[a,d]cycloheptene- 10-acetic acid; (S)-10,ll-Dihydro-3-[2-[6-(methylamino)pyridin-2-yl]-l-ethoxy]-5H-dibenzo[a,d]cycloheptene- 10-acetic acid; (S)-10,ll-Dihydro-3-[3-[4-(dimethylamino)pyridin-2-ylamino]-l-propyloxy]-5H-30 dibenzo[a,d]cycloheptene-10-acetic acid; (±)-10, ll-Dihydro-3-[3-[4-(ethylthio)pyridin-2-ylamino]-l-propy loxy ]-5H-dibenzo[a,d]cycloheptene-10-acetic acid; (S)-10,11 -Dihydro-3-[3-(4-chloropyridin-2-y lamino)-1 -propyloxy]-5H-dibenzo[a,d]cycloheptene- 10-acetic acid; 35 (±)-10,11 -Dihydro-2-methy l-3-[3-(pyridin-2-y lamino)-1 -propyloxy]-5H- dibenzo[a,d]cycloheptene- 10-acetic acid; -85- 011341 (S)-10,1 l-Dihydro-3-[3-(4-aminopyridin-2-ylarnino)-l-propy!oxy]-5K-dibenzo[a,d]cycloheptene- 10-acetic acid; (±)-10,11 -Dihydro-3-[3-(4-methylpyridin-2-ylamino)-1 -propyloxyj-dibenzo[b,f]oxepine-10-acetic acid; 5 (±)-10,11 -Dihydro-3-[2-[6-(methy lamino)pyridin-2-yl]-1 -ethoxy ]- dibenzo[b,f]oxepine-10-acetic acid; or (S)-10,11 -Dihydro-3-[3-(2-aminopyridin-4-y 1)-1 -propy loxy]-5H-dibenzo[a,i/]cycloheptene-10-acetic acid;or a pharmaceutically acceptable" sait thereof. 10
- 8. A pharmaceutical composition which comprises a compound accordïng toclaim 1 and a pharmaceutically acceptable carrier.
- 9. A pharmaceutical composition which comprises a compound accordïng to15 claim 1, an antineoplastic agent and a pharmaceutically acceptable carrier.
- 10. The pharmaceutical composition accordïng to claim 9 wherein theantineoplastic agent is topotecan.
- 11. The pharmaceutical composition accordïng to claim 9 wherein the antineoplastic agent is cisplatin.
- 12. A method of treating a disease State in which antagonism of the αγββreceptor is indicated which comprises administering to a subject in need thereof a 25 compound accordïng to claim 1.
- 13. A method of treating a disease State in which antagonism of the avB5receptor is indicated which comprises administering to a subject in need thereof acompound accordïng to claim 1. 30
- 14. A method of treating osteoporosis which comprises administering to asubject in need thereof a compound accordïng to claim 1.
- 15. A method for inhibiting angiogenesis which comprises administering to a35 subject in need thereof a compound accordïng to claim 1. -86- 011341
- 16. A method for inhibiting tumor growth or tumor metastasis which comprisesadministering to a subject in need thereof a compound according to claim 1.
- 17. A method of treating atherosclerosis or restenosis which comprisesadministering to a subject in need thereof a compound according to claim 1.
- 18. A method of treating inflammation which comprises administering to asubject in need thereof a compound according to claim 1.
- 19. A method of inhibiting tumor growth which comprises administeringstepwise or in physical combination a compound according to claim 1 and an antineoplasticagent.
- 20. The method according to claim 19 wherein the antineoplastic agent is15 topotecan.
- 21. The method according to claim 19 wherein the antineoplastic agent iscisplatin. 20(Π) wherein: 25 A is CH2 or O; R1 is H, halo or Cj.galkyl; R2 is H, Ci.6alkyl or CH2NR"R";X is O or CH2; Y-is 30-87- 01 1341 10 15 20G is NR", S or O; R’ is H, Ci-6alkyl, OCi-6alkyl, SCi-ôalkyl, NR"R" or halo;each R" independentlÿ is H or Cj^alkyl; ands is 0, 1 or 2; or a pharmaceutically acceptable sait thereof.(ΠΙ) wherein: A is CH2 or O; R* is H, halo or Cj.galkyl; R2 is H, Ci_6alkyl or CH2NR"R"; X is O or CH2; R’ is H, Ci.6alkyl, OC1.6alkyl, SCi.6alkyl, NR"R" or halo; andeach R" independentlÿ is H or Cj.galkyl;or a pharmaceutically acceptable sait thereof.
- 24, A process for preparing a compound of the formula (I) as defined in claim1, which process comprises reacting a compound of formula (IV) with a compound offormula (V):wherein Rl, R^, Y and A are as defined in formula (I), with any reactive functionalgroups protected, and L1 is OH or halo; and thereafter removing any protecting groups, and optionally forming apharmaceutically acceptable sait.
- 25. A process for preparing a compound of the formula (I) as defined in claim1, which process comprises reacting a compound of formula (IV) with a compound of 10 formula (VI):O-R’ (VI) 15 wherein Rl, R^, R’, R" and A are as defined in formula (I), with any reactive functional groups protected; and thereafter removing any protecting groups, and optionally forming apharmaceutically acceptable sait.
- 26. A process for preparing a compound of the formula (I) as defined in claim1, which process comprises reacting a compound of formula (IV) with a compound offormula (VII):(IV) (VII) 25 011341 -89- wherein Rl, r2, R" and A are as defined in formula (I), with any reactivefunctional groups protected; and thereafter removing any protecting groups, and opîionaiiy forming apharmaceutically acceptable sait. 5
- 27. A compound according to any one of daims 1 to 7 for use as amédicament.
- 28. The use of a compound of the formula (I) as defined in claim 1 in the10 manufacture of a médicament for the treatment of diseases in which antagonism of the ανβ3 receptor is indicated.
- 29. The use of a compound of the formula (I) as defined in claim 1 in themanufacture of a médicament for the treatment of diseases in which antagonism of the 15 av6g receptor is indicated.
- 30. The use of a compound of the formula (I) as defined in claim 1 in themanufacture of a médicament for the treatment of osteoporosis.
- 31. The use of a compound of the formula (I) as defined in claim 1 in the manufacture of a médicament for the inhibition of angiogenesis.
- 32. The use of a compound of the formula (I) as defined in claim 1 in themanufacture of a médicament for the inhibition of tumor growth or tumor metastasis. 25
- 33. The use of a compound of the formula (I) as defined in claim 1 in themanufacture of a médicament for the treatment of atherosclerosis or restenosis.
- 34. The use of a compound of the formula (I) as defined in claim 1 in the30 manufacture of a médicament for the treatment of inflammation.
- 35. The use of a compound of the formula (I) as defined in claim 1 and anantineoplastic agent in the manufacture of a médicament for the inhibition of tumor growthin physical combination or for stepwise administration. 35
- 36. The use according to claim 35 wherein the antineoplastic agent istopotecan. -90- 011341 37. The use according to claim 35 wherein the antineoplastic agent is cisplatin.
- 38. The use of a compound of the formula (I) as defined in claim 1 and an5 inhibitor of bone résorption in the manufacture of a médicament for the treatment of osteoporosis in physical combination or for stepwise administration.
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| US5934297P | 1997-09-19 | 1997-09-19 | |
| US6343897P | 1997-10-29 | 1997-10-29 |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| OA11341A true OA11341A (en) | 2003-12-10 |
Family
ID=26738652
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| OA1200000079A OA11341A (en) | 1997-09-19 | 2000-03-16 | Vitronectin receptor antagonists. |
Country Status (26)
| Country | Link |
|---|---|
| EP (1) | EP1025090A4 (en) |
| JP (1) | JP2001517658A (en) |
| KR (1) | KR20010024141A (en) |
| CN (1) | CN1278250A (en) |
| AP (1) | AP2000001766A0 (en) |
| AR (1) | AR015446A1 (en) |
| AU (1) | AU738433B2 (en) |
| BG (1) | BG104314A (en) |
| BR (1) | BR9812340A (en) |
| CA (1) | CA2303487A1 (en) |
| CO (1) | CO5011087A1 (en) |
| DZ (1) | DZ2609A1 (en) |
| EA (1) | EA200000336A1 (en) |
| HU (1) | HUP0003641A3 (en) |
| ID (1) | ID24162A (en) |
| IL (1) | IL135028A0 (en) |
| MA (1) | MA26547A1 (en) |
| NO (1) | NO20001407D0 (en) |
| NZ (1) | NZ503389A (en) |
| OA (1) | OA11341A (en) |
| PE (1) | PE122699A1 (en) |
| PL (1) | PL339381A1 (en) |
| SK (1) | SK4082000A3 (en) |
| TR (1) | TR200000721T2 (en) |
| TW (1) | TW513303B (en) |
| WO (1) | WO1999015508A1 (en) |
Families Citing this family (18)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CO4920232A1 (en) * | 1997-01-08 | 2000-05-29 | Smithkline Beecham Corp | DIBENZO [A, D] CYCLLOHEPTANE ACETIC ACIDS WITH VITRONECTIN RECEPTOR ANTAGONIST ACTIVITY |
| WO2000046215A1 (en) | 1999-02-03 | 2000-08-10 | Merck & Co., Inc. | Benzazepine derivatives as alpha-v integrin receptor antagonists |
| DE19936780A1 (en) | 1999-08-09 | 2001-02-15 | Basf Ag | New integrin receptor antagonists |
| US6881736B1 (en) | 1999-09-07 | 2005-04-19 | Smithkline Beecham Corporation | Vitronectin receptor antagonists |
| EG24179A (en) * | 1999-09-07 | 2008-09-28 | Smithkline Beecham Corp | Vitronectin receptor antagonists |
| US6514964B1 (en) | 1999-09-27 | 2003-02-04 | Amgen Inc. | Fused cycloheptane and fused azacycloheptane compounds and their methods of use |
| FR2806082B1 (en) * | 2000-03-07 | 2002-05-17 | Adir | NOVEL BICYCLIC ANTAGONIST VITRONECTIN RECEPTOR COMPOUNDS, PROCESS FOR THEIR PREPARATION AND PHARMACEUTICAL COMPOSITIONS CONTAINING THEM |
| EP1289959A1 (en) | 2000-06-15 | 2003-03-12 | Pharmacia Corporation | Dihydrostilbene alkanoic acid derivatives useful as vitronectin antagonists |
| JP2004513953A (en) * | 2000-10-24 | 2004-05-13 | メルク エンド カムパニー インコーポレーテッド | Dibenzoxazepine αV integrin receptor antagonist |
| CN1247258C (en) | 2001-04-24 | 2006-03-29 | 默克专利有限公司 | Combination therapy using anti-angiogenic agents and tnfa |
| GB0215867D0 (en) * | 2002-07-09 | 2002-08-14 | Glaxosmithkline Spa | Novel method and compounds |
| US20040224986A1 (en) | 2002-08-16 | 2004-11-11 | Bart De Corte | Piperidinyl targeting compounds that selectively bind integrins |
| UA87854C2 (en) | 2004-06-07 | 2009-08-25 | Мерк Энд Ко., Инк. | N-(2-benzyl)-2-phenylbutanamides as androgen receptor modulators |
| EP1973569B1 (en) | 2006-01-18 | 2013-05-22 | Merck Patent GmbH | Specific therapy using integrin ligands for treating cancer |
| US20100069302A1 (en) | 2007-07-18 | 2010-03-18 | Stefan Krueger | Specific therapy and medicament using integrin ligands for treating cancer |
| JP2012528079A (en) | 2009-05-25 | 2012-11-12 | メルク パテント ゲゼルシャフト ミット ベシュレンクテル ハフツング | Continuous administration of integrin ligand to treat cancer |
| KR20160147007A (en) | 2014-05-30 | 2016-12-21 | 화이자 인코포레이티드 | Carbonitrile derivatives as selective androgen receptor modulators |
| WO2023275715A1 (en) | 2021-06-30 | 2023-01-05 | Pfizer Inc. | Metabolites of selective androgen receptor modulators |
Family Cites Families (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CH443281A (en) * | 1964-02-18 | 1967-09-15 | Hoffmann La Roche | Process for the preparation of dibenzocycloheptatriene compounds |
| US3335148A (en) * | 1966-02-17 | 1967-08-08 | Lilly Co Eli | 9(3-pyridyl)derivative of fluorene, 9-fluorenol, xanthene, 9-xanthenol and the corresponding nonphytotoxic acid addition salts thereof |
| US5698551A (en) * | 1995-04-07 | 1997-12-16 | Novo Nordisk A/S | Heterocyclic compounds |
| JP3960482B2 (en) * | 1995-06-29 | 2007-08-15 | スミスクライン・ビーチャム・コーポレイション | Integrin receptor antagonist |
| US5659033A (en) * | 1995-09-13 | 1997-08-19 | Neurogen Corporation | N-aminoalkylfluorenecarboxamides; a new class of dopamine receptor subtype specific ligands |
| WO1998015278A1 (en) * | 1996-10-07 | 1998-04-16 | Smithkline Beecham Corporation | Method for stimulating bone formation |
| CO4920232A1 (en) * | 1997-01-08 | 2000-05-29 | Smithkline Beecham Corp | DIBENZO [A, D] CYCLLOHEPTANE ACETIC ACIDS WITH VITRONECTIN RECEPTOR ANTAGONIST ACTIVITY |
-
1998
- 1998-09-16 DZ DZ980219A patent/DZ2609A1/en active
- 1998-09-16 AR ARP980104604A patent/AR015446A1/en not_active Application Discontinuation
- 1998-09-16 PE PE1998000881A patent/PE122699A1/en not_active Application Discontinuation
- 1998-09-18 AP APAP/P/2000/001766A patent/AP2000001766A0/en unknown
- 1998-09-18 JP JP2000512816A patent/JP2001517658A/en not_active Withdrawn
- 1998-09-18 EP EP98947116A patent/EP1025090A4/en not_active Withdrawn
- 1998-09-18 ID IDW20000468A patent/ID24162A/en unknown
- 1998-09-18 CO CO98053986A patent/CO5011087A1/en unknown
- 1998-09-18 WO PCT/US1998/019466 patent/WO1999015508A1/en not_active Application Discontinuation
- 1998-09-18 MA MA25266A patent/MA26547A1/en unknown
- 1998-09-18 HU HU0003641A patent/HUP0003641A3/en unknown
- 1998-09-18 SK SK408-2000A patent/SK4082000A3/en unknown
- 1998-09-18 AU AU93972/98A patent/AU738433B2/en not_active Ceased
- 1998-09-18 NZ NZ503389A patent/NZ503389A/en not_active IP Right Cessation
- 1998-09-18 KR KR1020007002901A patent/KR20010024141A/en not_active Application Discontinuation
- 1998-09-18 IL IL13502898A patent/IL135028A0/en unknown
- 1998-09-18 TR TR2000/00721T patent/TR200000721T2/en unknown
- 1998-09-18 EA EA200000336A patent/EA200000336A1/en unknown
- 1998-09-18 CN CN98810811A patent/CN1278250A/en active Pending
- 1998-09-18 CA CA002303487A patent/CA2303487A1/en not_active Abandoned
- 1998-09-18 BR BR9812340-8A patent/BR9812340A/en not_active Application Discontinuation
- 1998-09-18 PL PL98339381A patent/PL339381A1/en unknown
- 1998-09-23 TW TW087115535A patent/TW513303B/en not_active IP Right Cessation
-
2000
- 2000-03-16 OA OA1200000079A patent/OA11341A/en unknown
- 2000-03-17 NO NO20001407A patent/NO20001407D0/en not_active Application Discontinuation
- 2000-04-07 BG BG104314A patent/BG104314A/en unknown
Also Published As
| Publication number | Publication date |
|---|---|
| DZ2609A1 (en) | 2003-03-01 |
| ID24162A (en) | 2000-07-13 |
| AU9397298A (en) | 1999-04-12 |
| CO5011087A1 (en) | 2001-02-28 |
| AR015446A1 (en) | 2001-05-02 |
| PE122699A1 (en) | 2000-02-12 |
| WO1999015508A1 (en) | 1999-04-01 |
| KR20010024141A (en) | 2001-03-26 |
| TW513303B (en) | 2002-12-11 |
| PL339381A1 (en) | 2000-12-18 |
| BR9812340A (en) | 2001-12-18 |
| HUP0003641A3 (en) | 2002-10-28 |
| BG104314A (en) | 2001-01-31 |
| NO20001407L (en) | 2000-03-17 |
| EA200000336A1 (en) | 2000-10-30 |
| CN1278250A (en) | 2000-12-27 |
| HUP0003641A2 (en) | 2001-03-28 |
| CA2303487A1 (en) | 1999-04-01 |
| NO20001407D0 (en) | 2000-03-17 |
| AU738433B2 (en) | 2001-09-20 |
| IL135028A0 (en) | 2001-05-20 |
| SK4082000A3 (en) | 2000-09-12 |
| NZ503389A (en) | 2002-03-28 |
| EP1025090A1 (en) | 2000-08-09 |
| MA26547A1 (en) | 2004-12-20 |
| EP1025090A4 (en) | 2000-11-08 |
| JP2001517658A (en) | 2001-10-09 |
| AP2000001766A0 (en) | 1998-09-18 |
| TR200000721T2 (en) | 2000-11-21 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| AU733417B2 (en) | Vitronectin receptor antagonists | |
| US6191304B1 (en) | Vitronectin receptor antagonists | |
| OA11341A (en) | Vitronectin receptor antagonists. | |
| CA2304000A1 (en) | Vitronectin receptor antagonist | |
| CA2303846A1 (en) | Vitronectin receptor antagonist | |
| EP1146874B1 (en) | Vitronectin receptor antagonist | |
| NZ517296A (en) | Vitronectin receptor antagonists | |
| US6576643B2 (en) | Vitronectin receptor antagonists | |
| US20020019387A1 (en) | Vitronectin receptor antagonist | |
| MXPA00002700A (en) | Vitronectin receptor antagonists | |
| US20020040136A1 (en) | Vitronectin receptor antagonist | |
| CZ2000979A3 (en) | Antagonist of a vitronectin receptor |