Classifications

• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07D—HETEROCYCLIC COMPOUNDS
  • C07D267/00—Heterocyclic compounds containing rings of more than six members having one nitrogen atom and one oxygen atom as the only ring hetero atoms
  • C07D267/02—Seven-membered rings
  • C07D267/08—Seven-membered rings having the hetero atoms in positions 1 and 4
  • C07D267/12—Seven-membered rings having the hetero atoms in positions 1 and 4 condensed with carbocyclic rings or ring systems
  • C07D267/16—Seven-membered rings having the hetero atoms in positions 1 and 4 condensed with carbocyclic rings or ring systems condensed with two six-membered rings
  • C07D267/20—[b, f]-condensed
• • 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
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P27/00—Drugs for disorders of the senses
  • A61P27/02—Ophthalmic agents
• • 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
  • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
  • A61P31/12—Antivirals
• • 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

Definitions

• the present invention relates to dibenzo-azepine de ⁇ vatives, their synthesis, and their use as ⁇ v integ ⁇ n receptor antagonists. More particularly, the compounds of the present invention are antagonists of the integ ⁇ n receptors ccv ⁇ 3, ocv ⁇ 5, and/or ⁇ v ⁇ and are useful for inhibiting bone resorption, treating and preventing osteoporosis, and inhibiting vascular restenosis, diabetic retinopathy, macular degeneration, angiogenesis, atherosclerosis, inflammation, wound healing, viral disease, tumor growth, and metastasis.
• Integ ⁇ n receptors are heterodime ⁇ c transmembrane receptors through which cells attach and communicate with extracellular matnces and other cells.
• the compounds herein are useful for inhibiting bone resorption.
• Bone resorption is mediated by the action of cells known as osteoclasts.
• Osteoclasts are large mult ucleated cells of up to about 400 mm in diameter that resorb mineralized tissue, chiefly calcium carbonate and calcium phosphate, in vertebrates.
• Osteoclasts are actively motile cells that migrate along the surface of bone, and can bind to bone, secrete necessary acids and proteases, thereby causing the actual resorption of mineralized tissue from the bone. More specifically, osteoclasts are believed to exist in at least two physiological states, namely, the secretory state and the migratory or motile state.
• osteoclasts are flat, attach to the bone matrix via a tight attachment zone (sealing zone), become highly pola ⁇ zed, form a ruffled border, and secrete lysosomal enzymes and protons to resorb bone.
• the adhesion of osteoclasts to bone surfaces is an important initial step in bone resorption.
• osteoclasts migrate across bone matrix and do not take part in resorption until they again attach to bone ⁇ v Integ ⁇ ns are involved osteoclast attachment, activation and migration
• the most abundant ⁇ v integrin in osteoclasts e.g., in rat, chicken, mouse and human osteoclasts, is an integ ⁇ n receptor known as ⁇ v ⁇ 3, which is thought to interact in bone with mat ⁇ x proteins that contain the RGD sequence Antibodies to ⁇ v ⁇ 3 block bone resorption in vitro indicating that this integrin plays a key role in the resorptive process.
• osteoporosis hypercalcemia of malignancy
• osteopenia due to bone metastases
• pe ⁇ odontal disease hyperparathyroidism
• pe ⁇ articular erosions in rheumatoid arth ⁇ tis Paget's disease
• immobilization-induced osteopenia and glucocorticoid-induced osteoporosis. All of these conditions are characterized by bone loss, resulting from an imbalance between bone resorption, i.e. breakdown, and bone formation, which continues throughout life at the rate of about 14% per year on the average.
• the rate of bone turnover differs from site to site; for example, it is higher in the trabecular bone of the vertebrae and the alveolar bone in the jaws than in the cortices of the long bones.
• the potential for bone loss is directly related to turnover and can amount to over 5% per year vertebrae immediately following menopause, a condition which leads to increased fracture ⁇ sk.
• In the United States there are currently about 20 million people with detectable fractures of the vertebrae due to osteoporosis
• ⁇ v ⁇ 3 ligands have been found to be useful in treating and/or inhibiting restenosis, i.e. recurrence of stenosis after corrective surgery on the heart valve, atherosclerosis, diabetic retinopathy, macular degeneration, and angiogenesis, i.e. formation of new blood vessels, and inhibiting viral disease.
• ⁇ v ⁇ 3 antagonists which inhibit angiogenesis can be useful in the treatment of cancer by inhibiting tumor growth (See, e.g., Brooks et al., Cell, 79:1157-1164 (1994), which is incorporated by reference herein in its entirety).
• compounds of this invention can also inhibit neovascula ⁇ zation by acting as antagonists of the integ ⁇ n receptor, ⁇ v ⁇ 5.
• a monoclonal antibody for ⁇ v ⁇ 5 has been shown to inhibit VEGF-induced angiogenesis in rabbit cornea and the chick cho ⁇ oallantoic membrane model (See M.C F ⁇ edlander, et.al., Science 270: 1500-1502 (1995), which is incorporated by reference herein in its entirety).
• compounds that antagonize ⁇ v ⁇ 5 are useful for treating and preventing macular degeneration, diabetic retinopathy, tumor growth, and metastasis
• compounds of the instant invention can inhibit angiogenesis and inflammation by acting as antagonists of the integ ⁇ n receptor, ⁇ v ⁇ 6, which is expressed dunng the later stages of wound healing and remains expressed until the wound is closed (See Ch ⁇ stofidou-Solomidou, et al., "Expression and Function of Endothehal Cell ⁇ v Integ ⁇ n Receptors in Wound-Induced Human Angiogenesis in Human Skin/SCLD Mice Chimeras, Amencan Journal of Pathology, Vol. 151, No. 4, pp. 975-983 (October 1997), which is incorporated by reference herein in its entirety).
• ⁇ v ⁇ 6 plays a role in the remodeling of the vasculature du ⁇ ng the later stages of angiogenesis. Also, ⁇ v ⁇ 6 participates in the modulation of epithelial inflammation and is induced m response to local injury or inflammation (See Xiao-Zhu Huang, et al., "Inactivation of the Integ ⁇ n ⁇ 6 Subunit Gene Reveals a Role of Epithelial Integ ⁇ ns in Regulating Inflammation in the Lungs and Skin," Journal of Cell Biology, Vol. 133, No.4, pp 921-928 (May 1996), which is incorporated by reference herein in its entirety). Accordingly, compounds that antagonize ⁇ v ⁇ 6 are useful in treating or preventing cancer by inhibiting tumor growth and metastasis.
• certain compounds of this invention antagonize both the ⁇ v ⁇ 3 and ⁇ v ⁇ 5 receptors.
• These compounds referred to as “dual ⁇ v ⁇ 3/ ⁇ v ⁇ 5 antagonists,” are useful for inhibiting bone resorption, treating and preventing osteoporosis, and inhibiting vascular restenosis, diabetic retinopathy, macular degeneration, angiogenesis, atherosclerosis, inflammation, viral disease, tumor growth, and metastasis.
• certain compounds of this invention are useful as mixed ⁇ v ⁇ 3, ⁇ v ⁇ 5, and ⁇ v ⁇ receptor antagonists.
• Peptidyl as well as peptidomimetic antagonists of the ⁇ v ⁇ 3 integ ⁇ n receptor have been descnbed both in the scientific and patent literature For example, reference is made to W.J. Hoekstra and B.L. Poulter, Curr. Med. Chem.
• the ⁇ v ⁇ 3 integ ⁇ n receptor recognizes the Arg-Gly-Asp (RGD) t ⁇ peptide sequence in its cognate mat ⁇ x and cell surface glycoproteins (see J. Samanen, et al., "Vascular Indications for Integ ⁇ n ⁇ v Antagonists," Curr. Pharmaceut. Design 3: 545-584 (1997)).
• RGD Arg-Gly-Asp
• a benzazepine nucleus has been employed among others by Genentech and SmithKline Beecham as a conformationally constrained Gly-Asp mimetic to elaborate nonpeptide ⁇ v ⁇ 3 integ ⁇ n receptor antagonists substituted at the N-termmus with heterocyclic arginme mimetics (see R.M.
• Keenan et al "Discovery of Potent Nonpeptide Vitronectin Receptor ( ⁇ v ⁇ 3) Antagonists," J. Med. Chem. 40: 2289-2292 (1997); R.M. Keenan et al., "Benzimidazole De ⁇ vatives As Arginme Mimetics in 1,4-Benzod ⁇ azep ⁇ ne Nonpeptide Vitronectin Receptor ( ⁇ v ⁇ 3) Antagonists," Bioorg. Med. Chem. Lett. 8- 3165-3170 (1998); and R.M. Keenan et al., "Discovery of an Imidazopy ⁇ dine-
• Patents assigned to SmithKlme Beecham that disclose such benzazepme- based ⁇ v ⁇ 3 integ ⁇ n receptor antagonists include WO 96/00574, WO 96/00730, WO 96/06087, WO 96/26190, WO 97/24119, WO 97/24122, WO 97/24124, and WO 98/15278 and to Genentech include WO 97/34865.
• the dibenzocycloheptene nucleus has also been employed as a Gly-Asp mimetic to afford ⁇ v ⁇ 3 antagonists (see WO 97/01540 and WO 98/30542 both assigned to SmithKline Beecham).
• ⁇ v ⁇ 3 antagonists see WO 97/01540 and WO 98/30542 both assigned to SmithKline Beecham.
• ⁇ v ⁇ 3 antagonists see WO 97/01540 and WO 98/30542 both assigned to SmithKline Beecham.
• ⁇ v integrin receptor antagonists that display improved potency, pharmacodynamic, and pharmacokinetic properties, such as oral bioavailability and significant duration of action.
• Such compounds would prove to be useful for the treatment, prevention, or suppression of various pathologies enumerated above that are mediated by ⁇ v binding and cell adhesion and activation.
• the present invention relates to compounds of formula I
• U and V each independently represent N or CR6, provided that no more than one U represents N and no more than one V represents N;
• X is O; S(O)0-2; NR4; or CR1R2;
• Y is selected from the group consisting of
• Z is selected from the group consisting of
• a 5- or 6-membered monocychc aromatic or nonaromatic ⁇ ng system having 1, 2, 3 or 4 heteroatoms selected from the group consisting of N, O, and S wherein the non-aromatic ⁇ ng nitrogen atoms are unsubstituted or substituted with one R ' substituent and the ⁇ ng carbon atoms are unsubstituted or substituted with one or two R6 substituents, and
• a 9- to 14-membered polycychc ⁇ ng system wherein one or more of the ⁇ ngs is aromatic, and wherein the polycychc ⁇ ng system has 1, 2, 3 or 4 heteroatoms selected from the group consisting of N, O, and S, and wherein the non-aromatic ring nitrogen atoms are unsubstituted or substituted with one R substituent and the ⁇ ng carbon atoms are unsubstituted or substituted with one or two R" substituents;
• R! and R ⁇ are each independently selected from the group consisting of hydrogen, halo, Ci-8 alkyl, C 2 -8 alkenyl, C2-8 alkynyl, C3-8 cycloalkyl, C3-8 cycloheteroalkyl, C3-8 cycloalkyl Ci-6 alkyl,
• Ci-6 alkyl aryl, aryl C ⁇ . ⁇ alkyl, ammo C[. alkyl, Ci-4 acylamino Ci-6 alkyl,
• Ci-6 alkyl 1-2 amino Ci-6 alkyl, hydroxy C ⁇ _6 alkyl,
• Ci-6 alkylthio Ci-6 alkyl, hydroxycarbonyl C 1-6 alkyl,
• each R3 is independently selected from the group consisting of hydrogen, halo, aryl,
• Ci-6 alkylthio aryl Ci-6 alkylthio, aminocarbonyl,
• each R 4 is independently selected from the group consisting of hydrogen
• any of the alkyl groups of R 4 are either unsubstituted or substituted with one to three R ⁇ substituents;
• R5 is selected from the group consisting of hydrogen, Ci-8 alkyl, aryl, aryl C ⁇ - ⁇ alkyl,
• each R" is independently selected from the group consisting of hydrogen, halo,
• Ci-6 alkyl C3- cycloalkyl, C3-8 cycloheteroalkyl, C3-8 cycloalkyl Ci-6 alkyl, C3-8 cycloheteroalkyl Ci- alkyl, aryl, aryl Ci-6 alkyl, amino, amino C ⁇ _6 alkyl, Ci-4 acylammo, Ci-4 acylammo Ci- alkyl, (Ci-6 alkyl) 1-2 amino, (Ci- alkyl)i-2 ammo Ci- alkyl,
• Ci-6 alkyl-S(O)i- 2 Ci-6 alkyl-S(O)i- 2 , (Ci-6 alkyl) ⁇ _2 aminocarbonyl, Ci-6 alkyloxycarbonylamino, (C ⁇ _6 alkyl)i-2 aminocarbonyloxy, (aryl Ci- alky l-2 amino,
• R is selected from the group consisting of hydrogen, Ci-8 alkyl, C2-8 alkenyl, C 2 -8 alkynyl, C3-8 cycloalkyl,
• the present invention also relates to pharmaceutical compositions comp ⁇ sing the compounds of the present invention and a pharmaceutically acceptable earner
• the present invention also relates to methods for making the pharmaceutical compositions of the present invention
• the present invention also relates to methods for eliciting an ⁇ v integ ⁇ n receptor antagonizing effect in a mammal in need thereof by admmiste ⁇ ng the compounds and pharmaceutical compositions of the present invention
• the present invention also relates to methods for inhibiting bone resorption, restenosis, atherosclerosis, inflammation, viral disease, diabetic retinopathy, macular degeneration, angiogenesis, wound healing, tumor growth, and metastasis by admmiste ⁇ ng the compounds and pharmaceutical compositions of the present invention
• the present invention also relates to methods for treating osteoporosis by administering the compounds and pharmaceutical compositions of the present invention
• the present invention relates to compounds useful as ⁇ v integ ⁇ n receptor antagonists.
• Representative compounds of the present invention are descnbed by structural formula I:
• U and V each independently represent N or CR6, provided that no more than one U represents N and no more than one V represents N;
• X is O; S(O)0-2; NR 4 ; or CR1R2,
• Y is selected from the group consisting of
• any methylene (CH2) carbon atom in Y, other than in R 4 can be substituted by one or two R ⁇ substituents;
• Z is selected from the group consisting of
• a 9- to 14-membered polycychc nng system wherein one or more of the ⁇ ngs is aromatic, and wherein the polycychc nng system has 1, 2, 3 or 4 heteroatoms selected from the group consisting of N, O, and S, and wherein the non-aromatic nng nitrogen atoms are unsubstituted or substituted with one R substituent and the ⁇ ng carbon atoms are unsubstituted or substituted with one or two R ⁇ substituents;
• R! and R2 are each independently selected from the group consisting of hydrogen, halo,
• Ci-6 alkyl 1-2 amino C ⁇ _6 alkyl, hydroxy Ci-6 alkyl, Ci-6 alkoxy Ci-6 alkyl,
• each R3 is independently selected from the group consisting of hydrogen, halo, aryl, C 1-8 alkyl,
• each R 4 is independently selected from the group consisting of hydrogen
• any of the alkyl groups of R4 are either unsubstituted or substituted with one to three R ⁇ substituents;
• R5 is selected from the group consisting of hydrogen
• each R6 is independently selected from the group consisting of hydrogen, halo,
• R IS selected from the group consisting of hydrogen, Ci-8 alkyl, C2-8 alkenyl, C2-8 alkynyl, C3-8 cycloalkyl,
• C3-8 cycloheteroalkyl C3-8 cycloalkyl Ci-6 alkyl, C3-8 cycloheteroalkyl Ci-6 alkyl, arylCi-6 alkyl, arylcarbonyl,
• U and V are preferably CR6
• X is preferably oxygen or sulfur. More preferably X is oxygen.
• Y is preferably selected from the group consisting of
• Y is selected from the group consisting of
• Y is preferably selected from the group consisting of
• any methylene (CH 2 ) carbon atom in Y, other than in R 4 can be substituted by one or two R ⁇ substituents.
• Z is preferably is preferably selected from the group consisting of
• R7 is as defined above and the ⁇ ng carbon atoms are unsubstituted or substituted with one or two R6 substituents as defined above More preferably Z is selected from the group consisting of
• R7 is as defined above and the ring carbon atoms are unsubstituted or substituted with one or two R6 substituents as defined above Most preferably Z is
• R7 is as defined above and the ring carbon atoms are unsubstituted or substituted with one or two R6 substituents as defined above.
• R! and R2 are preferably independently selected from the group consisting of hydrogen, C 1-6 alkyl, C3-8 cycloalkyl, C3-8 cycloheteroalkyl, and aryl -3 alkyl.
• R* and R 2 are independently selected from hydrogen and C 1-3 alkyl.
• each R ⁇ is preferably selected from the group consisting of hydrogen, aryl,
• R3 is hydrogen or oxo.
• each R 4 is preferably selected from the group consisting of hydrogen, C 1-8 alkyl, aryl Ci- alkyl, C3_8 cycloalkyl, Ci-4 alkoxy Ci- alkyl, Ci- alkylsulfonyl, arylCi-6 alkylsulfonyl,
• Ci-6 alkoxycarbonyl aryl Ci-6 alkoxycarbonyl, C ⁇ _6 alkylcarbonyl, arylcarbonyl, aryl C 1-6 alkylcarbonyl,
• R 4 is selected from the group consisting of hydrogen
• Ci-4 alkoxy Ci-4 alkyl Most preferably R4 is hydrogen or methyl.
• R ⁇ is preferably selected from the group consisting of hydrogen, methyl, and ethyl. More preferably, R ⁇ is hydrogen.
• each R6 is preferably selected from the group consisting of hydrogen, cyano, halo,
• R7 is preferably hydrogen, Ci-3 alkyl, or aryl C ⁇ _3 alkyl. More preferably R7 IS hydrogen.
• salts of the compounds of this invention refer to non-toxic “pharmaceutically acceptable salts.”
• Other salts may, however, be useful in the preparation of the compounds according to the invention or of their pharmaceutically acceptable salts.
• Salts encompassed within the term "pharmaceutically acceptable salts" refer to non-toxic salts of the compounds of this invention which are generally prepared by reacting the free base with a suitable organic or inorganic acid.
• Representative salts include the following: acetate, benzenesulfonate, benzoate, bicarbonate, bisulfate, bitartrate, borate, bromide, calcium, camsylate, carbonate, chloride, clavulanate, citrate, dihydrochloride, edetate, edisylate, estolate, esylate, fumarate, gluceptate, gluconate, glutamate, glycollylarsanilate, hexylresorcinate, hydrabamine, hydrobromide, hydrochloride, hydroxynaphthoate, iodide, isothionate, lactate, lactobionate, laurate, malate, maleate, mandelate, mesylate, methylbromide, methylnitrate, methylsulfate, mucate, napsylate, nitrate, N-methylglucamine ammonium salt, oleate, oxalate, pam
• suitable pharmaceutically acceptable salts thereof may include alkali metal salts, e.g., sodium or potassium salts; alkaline earth metal salts, e.g., calcium or magnesium salts; and salts formed with suitable organic ligands, e.g., quaternary ammonium salts.
• the compounds of the present invention can have chiral centers and can thus occur as racemates, racemic mixtures, single enantiomers, diastereomeric mixtures, and individual diastereomers, with all isomeric forms being included in the present invention. Therefore, where a compound is chiral, the separate enantiomers or diastereomers, substantially free of the other, are included within the scope of the invention; further included are all mixtures of the two enantiomers.
• tautomers Some of the compounds described herein may exist with different points of attachment of hydrogen, referred to as tautomers. Such an example may be a ketone and its enol form, known as keto-enol tautomeis The individual tautomers as well as mixtures thereof are encompassed within the compounds of the present invention
• Compounds of the present invention may be separated into diastereoisomeric pairs of enantiomers by, for example, fractional crystallization from a suitable solvent, for example, methanol or ethyl acetate or a mixture thereof
• a suitable solvent for example, methanol or ethyl acetate or a mixture thereof
• the pair of enantiomers thus obtained may be separated into individual stereoisomers by conventional means, for example, by the use of an optically active acid as a resolving agent, or by HPLC using a chiral stationary phase
• any enantiomer of a compound of the present invention may be obtained by stereospecific synthesis using optically pure starting materials or reagents of known configuration.
• polymorphs and hydrates of the compounds of the instant invention are also included within the scope of the invention.
• the present invention includes within its scope prodrugs of the compounds of this invention
• prodrugs will be functional de ⁇ vatives of the compounds of this invention which are readily convertible in vivo into the required compound.
• the term "admiwel ⁇ ng” shall encompass the treatment of the va ⁇ ous conditions descnbed with the compound specifically disclosed or with a compound which may not be specifically disclosed, but which converts to the specified compound in vivo after administration to the patient.
• Conventional procedures for the selection and preparation of suitable prodrug de ⁇ vatives are descnbed, for example, in "Design of Prodrugs," ed H. Bundgaard, Elsevier, 1985, which is incorporated by reference herein in its entirety.
• Metabolites of these compounds include active species produced upon introduction of compounds of this invention into the biological milieu
• the term "therapeutically effective amount” shall mean that amount of a drug or pharmaceutical agent that will elicit the biological or medical response of a tissue, system, animal or human that is being sought by a researcher or clinician.
• ⁇ v integ ⁇ n receptor antagonist refers to a compound which binds to and antagonizes either the ⁇ v ⁇ 3 receptor, the ⁇ v ⁇ 5 receptor, or the ⁇ v ⁇ receptor, or a compound which binds to and antagonizes combinations of these receptors (for example, a dual ⁇ v ⁇ 3/ ⁇ v ⁇ 5 receptor antagonist)
• alkyl shall mean straight or branched chain alkanes of one to ten total carbon atoms, or any number within this range (i.e., methyl, ethyl, 1- propyl, 2-propyl, n-butyl, s-butyl, t-butyl, etc.)
• alkenyl shall mean straight or branched chain alkenes of two to ten total carbon atoms, or any number within this range
• alkynyl shall mean straight or branched chain alkynes of two to ten total carbon atoms, or any number within this range
• cycloalkyl shall mean cyclic ⁇ ngs of alkanes of three to eight total carbon atoms, or any number within this range (i.e., cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl or cyclooctyl).
• cycloheteroalkyl shall mean a 3- to 8- membered fully saturated heterocyclic nng containing one or two heteroatoms chosen from N, O, or S.
• cycloheteroalkyl groups include, but are not limited to, pipe ⁇ dmyl, pyrro dinyl, azetidinyl, morphohnyl, and piperazinyl.
• alkoxy refers to straight or branched chain alkoxides of the number of carbon atoms specified (e.g., C ⁇ _5 alkoxy), or any number within this range (i.e., methoxy, ethoxy, etc.).
• alkylthio refers to straight or branched chain alkylsulfides of the number of carbon atoms specified (e.g., -5 alkylthio), or any number withm this range (i.e., methylthio, ethylthio, etc.).
• aryl refers to a monocychc or polycychc system comp ⁇ sing at least one aromatic ring, wherein the monocy c or polycychc system contains 0, 1, 2, 3, or 4 heteroatoms chosen from N, O, or S, and wherein the monocyhc or polycyhc system is either unsubstituted or substituted with one or more groups independently selected from hydrogen, halogen, Ci-io alkyl, C3-8 cycloalkyl, aryl, aryl Ci-8 alkyl, ammo, ammo-Ci-8 alkyl, Ci-3 acylammo, Ci-3 acylamino-Ci- 8 alkyl, Ci-6 alkylammo, Ci-6 alkylamino Ci-8 alkyl, Ci-6 dialkylamino, Ci-6 d ⁇ alkylam ⁇ no-Ci-8 alkyl, Cj-4 alkoxy, C ⁇ _4 alkoxy C -6 alkyl, Ci-4 alky
• aryl examples include, but are not limited to, phenyl, naphthyl, py ⁇ dyl, quinolyl, pyrrolyl, pyrazolyl, pyrazinyl, pynmidyl, lmidazolyl, tetrazolyl, benzofuranyl, benzothiophenyl, benzimidazolyl, benzthiazolyl, benzoxazolyl, mdolyl, thienyl, furyl, dihydrobenzofuryl, benzo(l,3) dioxolane, oxazolyl, isoxazolyl, oxadiazolyl, thiazolyl, and isothiazolyl, which are either unsubstituted or substituted with one or more groups independently selected from hydrogen, halogen, Ci_io alkyl, C3-8 cycloalkyl, aryl, aryl Ci-8 alkyl, amino, amino Ci-8 alkyl, Ci-3
• the aryl group is unsubstituted, mono-, di- or tn-substituted with one to three of the above-named substituents; most preferably, the aryl group is unsubstituted, mono- or di-substituted with one to two of the above-named substituents
• alkyl or aryl or either of their prefix roots appears in a name of a substituent (e.g , aryl Ci-6 alkyl), it shall be interpreted as including those limitations given above for "alkyl” and "aryl.”
• Designated numbers of carbon atoms e.g., Ci-8 shall refer independently to the number of carbon atoms in an alkyl or cyclic alkyl moiety or to the alkyl portion of a larger substituent in which alkyl appears as its prefix root.
• arylalkyl and “alkylaryl” include an alkyl portion where alkyl is as defined above and to include an aryl portion where aryl is as defined above
• arylalkyl include, but are not limited to, benzyl, fluorobenzyl, chlorobenzyl, phenylethyl, phenylpropyl, fluorophenylethyl, chlorophenylethyl, thienylmethyl, thienylethyl, and thienylpropyl.
• alkylaryl examples include, but are not limited to, toluene, ethylbenzene, propylbenzene, methylpy ⁇ dine, ethylpy ⁇ dine, propylpy ⁇ dine and butylpy ⁇ dme.
• two R! substituents when on the same carbon atom, can be taken together with the carbon atom to which they are attached to form a carbonyl group
• two R ⁇ substituents when on the same carbon atom, can be taken together with the carbon atom to which they are attached to form a carbonyl group.
• the limitation, that in the resultant compound the carbon atom or atoms at which R3 IS attached is itself attached to no more than one heteroatom, does not apply
• two R ⁇ substituents, when on the same carbon atom can be taken together with the carbon atom to which they are attached to form a cyclopropyl group
• halo shall include lodo, bromo, chloro, and fluoro.
• oxy means an oxygen (O) atom.
• thio means a sulfur (S) atom.
• O oxygen
• S sulfur
• N nitrogen.
• substituted shall be deemed to include multiple degrees of substitution by a named substitutent. Where multiple substituent moieties are disclosed or claimed, the substituted compound can be independently substituted by one or more of the disclosed or claimed substituent moieties, singly or plurally. By independently substituted, it is meant that the (two or more) substituents can be the same or different
• Ci-5 alkylcarbonylammo Ci- alkyl substituent is equivalent to
• va ⁇ ous substituents i.e. X, Y, Z, R , R2, R3 ? R4 anc ⁇ R5, and the subsc ⁇ pts m, n, and p are to be chosen in conformity with well-known pnnciples of chemical structure connectivity
• Representative compounds of the present invention typically display submicromolar affinity for the integ ⁇ n receptors, particularly the ⁇ v ⁇ 3, ⁇ v ⁇ 5, and/or ⁇ v ⁇ receptors.
• Compounds of this invention are therefore useful for treating mammals suffering from a bone condition caused or mediated by increased bone resorption, who are in need of such therapy.
• Pharmacologically effective amounts of the compounds, including pharamaceutically acceptable salts thereof, are administered to the mammal, to inhibit the activity of mammalian osteoclasts.
• the compounds of the present invention are administered in dosages effective to antagonize the ⁇ v ⁇ 3 receptor where such treatment is needed, as, for example, in the prevention or treatment of osteoporosis.
• the ⁇ v mteg ⁇ n receptor antagonizing effect is an ⁇ v ⁇ 3 antagonizing effect
• An illustration of the invention is the method wherein the ⁇ v ⁇ 3 antagonizing effect is selected from inhibition of bone resorption, restenosis, angiogenesis, diabetic retinopathy, macular degeneration, inflammation, viral disease, tumor growth, or metastasis.
• the ⁇ v ⁇ 3 antagonizing effect is the inhibition of bone resorption
• an example of the invention is the method wherein the ⁇ v integnn receptor antagonizing effect is an ⁇ v ⁇ 5 antagonizing effect. More specifically, the ⁇ v ⁇ 5 antagonizing effect is selected from inhibition of restenosis, angiogenesis, diabetic retinopathy, macular degeneration, inflammation, tumor growth, or metastasis
• Illustrating the invention is the method wherein the ⁇ v integ ⁇ n receptor antagonizing effect is a dual ⁇ v ⁇ 3/ ⁇ v ⁇ 5 antagonizing effect. More particularly, the dual ⁇ v ⁇ 3/ ⁇ v ⁇ 5 antagonizing effect is selected from inhibition of bone resorption, restenosis, angiogenesis, diabetic retinopathy, macular degeneration, inflammation, viral disease, tumor growth, or metastasis.
• Illustrating the invention is the method wherein the ⁇ v integ ⁇ n receptor antagonizing effect is an ⁇ v ⁇ antagonizing effect. More particularly, the ⁇ v ⁇ antagonizing effect is selected from inhibition of angiogenesis, inflammatory response, or wound healing
• Illustrating the invention is the method wherein the ⁇ v ⁇ 3 antagonizing effect is selected from inhibition of bone resorption, inhibition of restenosis, inhibition of angiogenesis, inhibition of diabetic retinopathy, inhibition of macular degeneration, inhibition of atherosclerosis, inflammation, viral disease, or inhibition of tumor growth and metastasis.
• the ⁇ v ⁇ 3 antagonizing effect is the inhibition of bone resorption.
• a pharmaceutical composition comp ⁇ smg any of the compounds descnbed above and a pharmaceutically acceptable earner is a pharmaceutical composition comp ⁇ smg any of the compounds descnbed above and a pharmaceutically acceptable earner.
• a pharmaceutical composition made by combining any of the compounds descnbed above and a pharmaceutically acceptable earner is a process for making a pharmaceutical composition compnsmg combining any of the compounds descnbed above and a pharmaceutically acceptable carrier
• the condition is selected from bone resorption, osteoporosis, restenosis, diabetic retinopathy, macular degeneration, angiogenesis, atherosclerosis, inflammation, viral disease, cancer, tumor growth, and metastasis More preferably, the condition is selected from osteoporosis and cancer. Most preferably, the condition is osteoporosis.
• the invention is a method of eliciting an ⁇ v integrin antagonizing effect in a mammal in need thereof, comp ⁇ sing admmiste ⁇ ng to the mammal a therapeutically effective amount of any of the compounds or any of the pharmaceutical compositions descnbed above.
• the ⁇ v integnn antagonizing effect is an ⁇ v ⁇ 3 antagonizing effect, more specifically the ⁇ v ⁇ 3 antagonizing effect is selected from inhibition of bone resorption, inhibition of restenosis, inhibition of atherosclerosis, inhibition of angiogenesis, inhibition of diabetic retinopathy, inhibition of macular degeneration, inhibition of inflammation, inhibition of viral disease, or inhibition of tumor growth or metastasis.
• the ⁇ v ⁇ 3 antagonizing effect is inhibition of bone resorption.
• the ⁇ v mtegnn antagonizing effect is an ⁇ v ⁇ 5 antagonizing effect, an ⁇ v ⁇ antagonizing effect, or a mixed ⁇ v ⁇ 3, ⁇ v ⁇ 5, and ⁇ v ⁇ antagonizing effect.
• Examples of ⁇ v ⁇ 5 antagonizing effects are inhibition of restenosis, atherosclerosis, angiogenesis, diabetic retinopathy, macular degeneration, inflammation, or tumor growth
• Examples of ⁇ v ⁇ antagonizing effects are inhibition of angiogenesis, inflammatory response, and wound healing
• Additional examples of the invention are methods of inhibiting bone resorption and of treating and/or preventing osteoporosis in a mammal in need thereof, comp ⁇ sing administe ⁇ ng to the mammal a therapeutically effective amount of any of the compounds or any of the pharmaceutical compositions decnbed above
• Additional illustrations of the invention are methods of treating hypercalcemia of malignancy, osteopenia due to bone metastases, pe ⁇ odontal disease, hyperparathyroidism, pe ⁇ articular erosions in rheumatoid arthntis, Paget's disease, immobilization-induced osteopenia, and glucocorticoid treatment in a mammal in need thereof, comp ⁇ sing administe
• Still further exemplifying the invention is the use of any of the compounds described above in the preparation of a medicament for the treatment and/or prevention of bone resorption, tumor growth, cancer, restenosis, atherosclerosis, diabetic retinopathy, macular degeneration, inflammation, viral disease, and/or angiogenesis.
• compositions further compnsmg an active ingredient selected from the group consisting of a ) an organic bisphosphonate or a pharmaceutically acceptable salt or ester thereof, b.) an estrogen receptor modulator, c ) a cytotoxic/antiprohferative agent, d.) a matnx metalloprotemase inhibitor, e ) an inhibitor of epi dermal-den ved, fibroblast-de ⁇ ved, or platelet- den ved growth factors, f.) an inhibitor of VEGF, g.) an inhibitor of Flk-1/KDR, Flt-1, Tck/T ⁇ e-2, or T ⁇ e-1, h.) a cathepsin K inhibitor, and l.) a prenylation inhibitor, such as a farnesyl transferase inhibitor or a geranylgeranyl transferase inhibitor or a dual farnesyl/geranylgeranyl transferase inhibitor; and mixtures thereof.
• an active ingredient selected from the group consisting
• the active ingredient is selected from the group consisting of: a ) an organic bisphosphonate or a pharmaceutically acceptable salt or ester thereof, b.) an estrogen receptor modulator, and c.) a cathepsin K inhibitor; and mixtures thereof.
• Nonhmiting examples of such bisphosphonates include alendronate, etidronate, pamidronate, nsedronate, ibandronate, and pharmaceutically acceptable salts and esters thereof.
• a particularly preferred bisphosphonate is alendronate, especially alendronate monosodium t ⁇ hydrate
• Nonhmitmg examples of estrogen receptor modulators include estrogen, progesterm, estradiol, droloxifene, raloxifene, and tamoxifene.
• Nonhmiting examples of cytotoxic/antiprohferative agents are taxol, vinc ⁇ stine, vinblastme, and doxorubicin.
• Cathepsin K formerly known as cathepsin 02, is a cysteme protease and is described in PCT International Application Publication No WO 96/13523, published May 9, 1996; U.S. Patent No. 5,501,969, issued March 3, 1996; and U.S Patent No. 5,736,357, issued Apnl 7, 1998, all of which are incorporated by reference herein in their entirety.
• Cysteine proteases specifically catheps s, are linked to a number of disease conditions, such as tumor metastasis, inflammation, arth ⁇ tis, and bone remodeling. At acidic pH's, cathepsms can degrade type-I collagen.
• Cathepsin protease inhibitors can inhibit osteoclastic bone resorption by inhibiting the degradation of collagen fibers and are thus useful in the treatment of bone resorption diseases, such as osteoporosis.
• the present invention is also directed to combinations of the compounds of the present invention with one or more agents useful in the prevention or treatment of osteoporosis.
• the compounds of the instant invention may be effectively administered in combination with effective amounts of other agents such as an organic bisphosphonate, an estrogen receptor modulator, or a cathepsin K inhibitor.
• Additional illustrations of the invention are methods of treating tumor growth or metastasis in a mammal in need thereof, comp ⁇ sing administe ⁇ ng to the mammal a therapeutically effective amount of a compound descnbed above and one or more agents known to be cytotoxic/antiprohferative. Also, the compounds of the present invention can be administered in combination with radiation therapy for treating tumor growth and metastasis.
• the integnn ⁇ v ⁇ 3 antagonist compounds of the present invention may be effectively administered in combination with a growth hormone secretagogue in the therapeutic or prophylactic treatment of disorders in calcium or phosphate metabolism and associated diseases.
• diseases include conditions which can benefit from a reduction in bone resorption.
• a reduction in bone resorption should improve the balance between resorption and formation, reduce bone loss or result in bone augmentation.
• a reduction in bone resorption can alleviate the pain associated with osteolytic lesions and reduce the incidence and/or growth of those lesions
• These diseases include: osteoporosis (including estrogen deficiency, immobilization, glucocorticoid-induced and senile), osteodystrophy, Paget ' s disease, myositis ossificans, Bechterew's disease, malignant hypercalcemia, metastatic bone disease, penodontal disease, cholelithiasis, nephro thiasis, urohthiasis, u ⁇ nary calculus, hardening of the artenes (sclerosis), arth ⁇ tis, bursitis, neuritis and tetany.
• osteoporosis including estrogen deficiency, immobilization, glucocorticoid-induced and senile
• osteodystrophy Paget ' s disease
• myositis ossificans Bechte
• ⁇ v ⁇ 3 receptor antagonist of the present invention and a growth hormone secretagogue, optionally including a third component comp ⁇ sing an organic bisphosphonate, preferably alendronate monosodium t ⁇ hydrate.
• the individual components of the combination can be administered separately at different times dunng the course of therapy or concurrently in divided or single combination forms.
• the instant invention is therefore to be understood as embracing all such regimes of simultaneous or alternating treatment, and the term "administe ⁇ ng" is to be interpreted accordingly. It will be understood that the scope of combinations of the compounds of this invention with other agents useful for treating integ ⁇ n -mediated conditions includes in p ⁇ nciple any combination with any pharmaceutical composition useful for treating osteoporosis.
• composition is intended to encompass a product compnsmg the specified ingredients in the specified amounts, as well as any product which results, directly or indirectly, from combination of the specified ingredients in the specified amounts.
• the compounds of the present invention can be administered in such oral dosage forms as tablets, capsules (each of which includes sustained release or timed release formulations), pills, powders, granules, elixirs, tinctures, suspensions, syrups and emulsions. Likewise, they may also be administered in intravenous (bolus or infusion), intrape ⁇ toneal, topical (e.g., ocular eyedrop), subcutaneous, intramuscular or transdermal (e.g., patch) form, all using forms well known to those of ordinary skill in the pharmaceutical arts. An effective but non-toxic amount of the compound desired can be employed as an ⁇ v ⁇ 3 antagonist.
• the dosage regimen utilizing the compounds of the present invention is selected in accordance with a variety of factors including type, species, age, weight, sex and medical condition of the patient; the seventy of the condition to be treated, the route of administration; the renal and hepatic function of the patient; and the particular compound or salt thereof employed.
• An ordinanly skilled physician, vete ⁇ na ⁇ an or clinician can readily determine and presc ⁇ be the effective amount of the drug required to prevent, counter or arrest the progress of the condition
• Oral dosages of the present invention when used for the indicated effects, will range between about 0.01 mg per kg of body weight per day (mg/kg/day) to about 100 mg/kg/day, preferably 0.01 to 10 mg/kg/day, and most preferably 0 1 to 5.0 mg/kg/day
• the compositions are preferably provided in the form of tablets containing 0.01, 0.05, 0 1, 0.5, 1.0, 2.5, 5.0, 10.0, 15.0, 25.0, 50.0, 100 and 500 milligrams of the active ingredient for the symptomatic adjustment of the dosage to the patient to be treated
• a medicament typically contains from about 0.01 mg to about 500 mg of the active ingredient, preferably, from about 1 mg to about 100 mg of active ingredient.
• the most preferred doses will range from about 0.1 to about 10 mg/kg/minute du ⁇ ng a constant rate infusion.
• compounds of the present invention may be administered in a single daily dose, or the total daily dosage may be administered m divided doses of two, three or four times daily
• preferred compounds for the present invention can be administered in mtranasal form via topical use of suitable intranasal vehicles, or via transdermal routes, using those forms of transdermal skm patches well known to those of ordinary skill in the art.
• the dosage administration will, of course, be continuous rather than intermittent throughout the dosage regimen
• the compounds herein descnbed in detail can form the active ingredient, and are typically administered m admixture with suitable pharmaceutical diluents, excipients or earners (collectively referred to herein as 'earner' matenals) suitably selected with respect to the intended form of administration, that is, oral tablets, capsules, elixirs, syrups and the like, and consistent with conventional pharmaceutical practices.
• the active drug component can be combined with an oral, non-toxic, pharmaceutically acceptable, inert earner such as lactose, starch, sucrose, glucose, methyl cellulose, magnesium stearate, dicalcium phosphate, calcium sulfate, mannitol, sorbitol and the like;
• an oral, non-toxic, pharmaceutically acceptable, inert earner such as lactose, starch, sucrose, glucose, methyl cellulose, magnesium stearate, dicalcium phosphate, calcium sulfate, mannitol, sorbitol and the like
• the oral drug components can be combined with any oral, non-toxic, pharmaceutically acceptable inert earner such as ethanol, glycerol, water and the hke
• suitable binders, lubricants, disintegrating agents and coloring agents can also be incorporated into the mixture.
• Suitable binders include starch, gelatin, natural sugars such as glucose or beta-lactose, corn sweeteners, natural and synthetic gums such as acacia, tragacanth or sodium alginate, carboxymethylcellulose, polyethylene glycol, waxes and the like
• Lub ⁇ cants used in these dosage forms include sodium oleate, sodium stearate, magnesium stearate, sodium benzoate, sodium acetate, sodium chloride and the like
• Disintegrators include, without limitation, starch, methyl cellulose, agar, bentonite, xanthan gum and the like.
• the compounds of the present invention can also be administered in the form of hposome delivery systems, such as small umlamellar vesicles, large unilamellar vesicles and multilamellar vesicles Liposomes can be formed from a va ⁇ ety of phosphohpids, such as cholesterol, stearylamme or phosphatidylcholines.
• Compounds of the present invention may also be delivered by the use of monoclonal antibodies as individual earners to which the compound molecules are coupled.
• the compounds of the present invention may also be coupled with soluble polymers as targetable drug earners.
• Such polymers can include polyvmylpyrrolidone, pyran copolymer, polyhydroxypropylmethacrylamide -phenol, polyhydroxy-ethylaspartamide-phenol, or polyethyleneoxide-polylysine substituted with palmitoyl residues.
• the compounds of the present invention may be coupled to a class of biodegradable polymers useful in achieving controlled release of a drug, for example, polylactic acid, polyglycohc acid, copolymers of polylactic and polyglycohc acid, polyepsilon caprolactone, polyhydroxy buty ⁇ c acid, polyorthoesters, polyacetals, polydihydropyrans, polycyanoacrylates and crosslinked or amphipathic block copolymers of hydrogels.
• a drug for example, polylactic acid, polyglycohc acid, copolymers of polylactic and polyglycohc acid, polyepsilon caprolactone, polyhydroxy buty ⁇ c acid, polyorthoesters, polyacetals, polydihydropyrans, polycyanoacrylates and crosslinked or amphipathic block copolymers of hydrogels.
• va ⁇ ous reagent symbols and abbreviations have the following meanings
• FABLRMS Fast-atom bombardment low-resolution mass spectrum.
• HPLC High Performance Liquid Chromatography.
• L1AIH4 Lithium aluminum chlonde.
• MgSO4 Magnesium sulfate.
• Na2C03 Sodium carbonate.
• Na2SO4 Sodium sulfate.
• NMM N-methylmorpholme. NMR: Nuclear magnetic resonance. NH4CI: Ammonium chlonde.
• Pd/C Palladium on activated carbon catalyst.
• Ph3P Tnphenylphosphine.
• novel compounds of the present invention can be prepared according to the procedures depicted below in Schemes 1 and 2 using appropriate starting matenals which are either available from commercial sources or readily made according to methods well-desc ⁇ bed in the published chemical literature.
• Alkylation of the seven- membered nng amide nitrogen is effected with an alkyl bromo- or lodoacetate the presence of an approp ⁇ ate base, such as sodium or potassium hydride, in a suitable solvent, such as tetrahydrofuran, N,N-d ⁇ methylformam ⁇ de, or dimethyl sulfoxide
• a suitable solvent such as tetrahydrofuran, N,N-d ⁇ methylformam ⁇ de, or dimethyl sulfoxide
• W is oxygen with a suitable reducing agent, such as lithium aluminum hyd ⁇ de, diborane, or borane-methyl sulfide in a suitable solvent, such as diethyl ether, tetrahydrofuran, or methylene chlonde.
• a suitable reducing agent such as lithium aluminum hyd ⁇ de, diborane, or borane-methyl sulfide in a suitable solvent, such as diethyl ether, tetrahydrofuran, or methylene chlonde.
• Z-Y represents Z-(CH2)m- -
• a phenolic intermediate such as L-5 (obtainable by demethylation of the anisole intermediate L4) using Mitsunobu-type conditions (t ⁇ phenylphosp e, dialkyl azodicarboxylate) (Synthesis 1981, 1-28; Organic Reactions 1992, 42, 335-656) in the presence of the alcohol reagent, Z(opt ⁇ onally protected)-(CH2)m- H.
• Z-Y represents Z-(CH2) ⁇ rS-
• a thiophenol precursor corresponding to L5 with an alkyl hahde such as Z(opt ⁇ onally protected)-(CH2)m-I> ⁇ n the presence of a suitable inorganic or organic base, followed by removal of any Z-protecting group and further manipulation, such as catalytic hydrogenation and/or removal of any C-termmus carboxyhc acid protecting group, such as by alkaline hydrolysis
• the thiophenol intermediate can be prepared from the phenol by means of the Newman-Karnes procedure Treatment of the Z(opt ⁇ onally protected)-(CH2)m-S- intermediate with one or two equivalents of a suitable oxidizing agent, such as MCPBA in methylene chlonde, and subsequent N- and C-termmi deprotection, give the compounds of the present invention wherein Z-Y represents Z-(CH 2 )m-S(O)- and Z-(CH 2 )m-S
• a phenolic intermediate such as 1 ⁇ 5
• benzoic acid derivative This can be accomplished, for example, by converting the phenol into its tnfluoromethanesulfonate ester followed by carbonyl insertion with carbon monoxide in the presence of potassium acetate, a palladium catalyst, such as palladium acetate, and l,l '-b ⁇ s(d ⁇ phenylphosph ⁇ no)ferrocene (dppf).
• the tnfluoromethanesulfonate ester can be converted into its benzomtnle de ⁇ vative with copper(I) cyanide in DMF, which is then hydrolyzed to the benzoic acid under standard conditions. Amide bond formation is then earned out using standard coupling reagents, such as BOP, under conditions well-known to practitioners of the art of synthetic organic chemistry. Final N- and C-termini deprotection affords the desired final compounds of the present invention.
• the compounds illustrated in the examples below are not to be construed as forming the only genus that is considered as the invention. They serve to further illustrate details for the preparation of the compounds of the present invention Those skilled in the art will readily understand that known va ⁇ ations of the conditions and processes of the following synthetic procedures can be used to prepare these compounds. All temperatures are degrees Celsius unless otherwise noted.
• Analytical and preparative HPLC was earned out using a Waters 600E Powerlme Multi Solvent Delivery System with 0.1 mL heads with a Rheodyne 7125 injector and a Waters 990 Photodiode Array Detector with a Gilson FC203 Microfraction collector.
• analytical and preparative HPLC a Vydac peptide -protein C-18 column, 4.6 x 250 mm was used with a C-18 Brownlee modular guard column.
• the acetonit ⁇ le used for the HPLC analyses was Fisher Optima grade.
• the HPLC radiodetector used was a Beckman 170 Radioisotope detector.
• osteoclasts When osteoclasts engage in bone resorption, they can cause the formation of pits in the surface of bone that they are acting upon Therefore, when testing compounds for their ability to inhibit osteoclasts, it is useful to measure the ability of osteoclasts to excavate these resorption pits when the inhibiting compound is present
• Consecutive 200 micron thick cross sections from a 6 mm cylinder of bovine femur diaphysis are cut with a low speed diamond saw (Isomet, Beuler, Ltd , Lake Bluff, II). Bone slices are pooled, placed in a 10% ethanol solution and ref ⁇ gerated until further use.
• bovme bone slices Prior to expe ⁇ mentation, bovme bone slices are ultrasonicated twice, 20 minutes each in H2O. Cleaned slices are placed in 96 well plates such that two control lanes and one lane for each drug dosage are available. Each lane represents either tnphcate or quadruplicate cultures.
• the bone slices in 96 well plates are ste ⁇ hzed by UV irradiation. P ⁇ or to incubation with osteoclasts, the bone slices are hydrated by the addition of 0.1 ml ⁇ MEM, pH 6.9 containing 5% fetal bovine serum and 1% penicillin/streptomycin.
• a cell suspension consisting of 5 x 106/ml in ⁇ MEM containing 5% fetal bovine serum, 10 nM l,25(OH)2D3, and pencilhn-streptomycm is prepared. 200 ml aliquots are added to bovine bone slices (200 mm x 6 mm) and incubated for 2 hrs. at 37°C in a humidified 5% CO2 atmosphere. The medium is removed gently with a micropipettor and fresh medium containing test compounds is added. The cultures are incubated for 48 hrs., and assayed for c-telopeptide (fragments of the al chain of type I collagen) by Crosslaps for culture media (Herlev, Denmark).
• Bovine bone slices are exposed to osteoclasts for 20-24 hrs and are processed for staining. Tissue culture media is removed from each bone slice Each well is washed with 200 ml of H2O, and the bone slices are then fixed for 20 minutes in 2.5% glutaraldehyde, 0.1 M cacodylate, pH 7.4 After fixation, any remaining cellular debris is removed by 2 min. ultrasonication in the presence of 0.25 M NH4OH followed by 2 X 15 mm ultrasonication in H2O.
• the bone slices are immediately stained for 6-8 min with filtered 1% toluidine blue and 1% borax After the bone slices have dned, resorption pits are counted in test and control slices Resorption pits are viewed in a Microphot Fx (Nikon) fluorescence microscope using a polanzing Nikon IGS filter cube. Test dosage results are compared with controls and resulting IC50 values are determined for each compound tested The appropriateness of extrapolating data from this assay to mammalian (including human) disease states is supported by the teaching found in Sato, M., et al., Journal of Bone and Mineral Research, Vol. 5, No. l, pp.
• TBS buffer 50 mM Tris-HCl pH 7.2, 150 mM NaCl, 1% BSA, 1 mM CaC 12, 1 mM MgCt ⁇ ).
• Optiplate PACKARD
• Pu ⁇ fied integ ⁇ n receptor ⁇ v ⁇ 3 was punfied from 293 cells overexpressing ⁇ v ⁇ 3 (Duong et al., J. Bone Mm. Res., 8:S378, 1993) according to Pytela (Methods in Enzymology, 144:475,
• Binding buffer 50 mM HEPES, pH 7.8, 100 mM NaCl, 1 mM Ca 2+ Mg 2 +, 0.5 mM PMSF
• Osteoblast-hke cells (1.8 cells), originally derived from mouse calva ⁇ a, were plated in CORNF G 24 well tissue culture plates in ⁇ MEM medium containing nbo- and deoxy ⁇ bonucleosides, 10% fetal bovine serum and penicilhn- streptomycin. Cells were seeded at 40,000/well in the morning. In the afternoon, bone marrow cells were prepared from six week old male Balb/C mice as follows-
• mice were sacrificed, tibiae removed and placed in the above medium. The ends were cut off and the marrow was flushed out of the cavity into a tube with a 1 mL sy ⁇ nge with a 27.5 gauge needle. The marrow was suspended by pipetting up and down. The suspension was passed through > 100 mm nylon cell strainer. The resulting suspension was centrifuged at 350 x g for seven minutes. The pellet was resuspended, and a sample was diluted in 2% acetic acid to lyse the red cells. The remaining cells were counted in a hemacytometer The cells were pelleted and resuspended at 1 x 10" cells/mL.
• Attachment medium HBSS with 1 mg/ml heat-mactivated fatty acid free BSA and 2 mM CaCl2 3.
• Glucosaminidase substrate solution 3.75 mM p-mtrophenyl N-acetyl-beta-
• D-glucosaminide 0.1 M sodium citrate, 0.25% Triton, pH 5.0. 4 Glycine-EDTA developing solution: 50 mM glycine, 5 mM EDTA, pH 10.5.
• Non-adherent cells were removed by gently washing the plates (3X) with DPBS and then incubated with glucosaminidase substrate solution (100 ⁇ l/well), overnight at room temperature in the dark.
• glucosaminidase substrate solution 100 ⁇ l/well
• standard curve of glucosaminidase activity was determined for each expenment by adding samples of cell suspension directly to wells containing the enzyme substrate solution. 5. The next day, the reaction was developed by addition of 185 ⁇ l/well of glycine/EDTA solution and reading absorbance at 405 nm using a Molecular Devices V-Max plate reader. Average test absorbance values (4 wells per test samples) were calculated. Then, the number of attached cells at each drug concentration was quantitated versus the standard curve of cells using the Softmax program.
• an oral composition 100 mg of any of the compounds of the present invention are formulated with sufficient finely divided lactose to provide a total amount of 580 to 590 mg to fill a size O hard gel capsule
• Representative compounds of the present invention were tested and found to bind to human ⁇ v ⁇ 3 integ ⁇ n. These compounds were generally found to have IC50 values less than about 100 nM in the SPA assay. Representative compounds of the present invention were tested and generally found to inhibit > 50% the attachment of ⁇ v ⁇ 5 expressing cells to plates coated with vitronectin at concentrations of about 1 ⁇ M.
• vanous changes, modifications and substitutions can be made therein without departing from the spi ⁇ t and scope of the invention.
• effective dosages other than the preferred doses as set forth hereinabove may be applicable as a consequence of va ⁇ ations in the responsiveness of the mammal being treated for seventy of bone disorders caused by resorption, or for other indications for the compounds of the invention indicated above.

Landscapes

• Health & Medical Sciences (AREA)
• Organic Chemistry (AREA)
• Chemical & Material Sciences (AREA)
• Life Sciences & Earth Sciences (AREA)
• General Chemical & Material Sciences (AREA)
• Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
• Medicinal Chemistry (AREA)
• Pharmacology & Pharmacy (AREA)
• Chemical Kinetics & Catalysis (AREA)
• Animal Behavior & Ethology (AREA)
• General Health & Medical Sciences (AREA)
• Public Health (AREA)
• Veterinary Medicine (AREA)
• Bioinformatics & Cheminformatics (AREA)
• Engineering & Computer Science (AREA)
• Rheumatology (AREA)
• Cardiology (AREA)
• Heart & Thoracic Surgery (AREA)
• Physical Education & Sports Medicine (AREA)
• Oncology (AREA)
• Communicable Diseases (AREA)
• Vascular Medicine (AREA)
• Pain & Pain Management (AREA)
• Urology & Nephrology (AREA)
• Ophthalmology & Optometry (AREA)
• Orthopedic Medicine & Surgery (AREA)
• Virology (AREA)
• Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
• Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
• Plural Heterocyclic Compounds (AREA)

Priority Applications (4)

Applications Claiming Priority (2)

Publications (1)

Family

ID=22391134

Family Applications (1)

Country Status (5)

Cited By (5)

Citations (3)

Family Cites Families (1)

• 2000
  • 2000-02-14 AU AU33643/00A patent/AU750584B2/en not_active Ceased
  • 2000-02-14 CA CA002362334A patent/CA2362334A1/en not_active Abandoned
  • 2000-02-14 WO PCT/US2000/003796 patent/WO2000048603A1/en not_active Application Discontinuation
  • 2000-02-14 JP JP2000599395A patent/JP2002537260A/ja not_active Withdrawn
  • 2000-02-14 EP EP00911811A patent/EP1169042A4/en not_active Withdrawn

Patent Citations (3)

Non-Patent Citations (2)

Also Published As

Similar Documents

Legal Events