Inhibiting osteoclast-mediated bone resorption using substituted phenyl derivatives

Abstract: Compounds of the general formula Y is an integer of from 0 to 6 is chosen from: 0, SO₂, -CONH-, -NHCO-, -CH₂- or R¹ is substituted or unsubstituted mono- or polycyclic saturated hetero-cyclic ring system having 1, 2 or 3 heteroatoms where said heteroatoms are independently chosen from N, O and S and said substituents are chosen from the group comprising R²; R² is C₁₋₆ alkyl, aryl C₀₋₆alkyl, hydroxy C₀₋₆alkyl, C₁₋₆ alkoxy C₀₋₆alkyl, carboxy C₀₋₆alkyl oxo, halogen, CF₃, C₀₋₄alkylamino-C₀₋₆alkyl or C₀₋₄dialkylamino-C₀₋₆alkyl; R³ is C₁₋₈ alkyl or cycloalkyl, aryl C₀₋₄alkyl, hydroxy C₀₋₄alkyl C₁₋₄alkyloxy C₀₋₄alkyl, carboxy C₀₋₄alkyl, halogen, CF₃ or hydrogen; R⁴ is hydrogen, C₁₋₆alkyl, aryl C₀₋₄alkyl or C₁₋₆alkylcarbonyloxymethyl; and R⁵ is C₁₋₆alkyl, aryl C₀₋₄alkyl or heterocycly C₀₋₄alkyl and the pharmaceutically acceptable salts thereof; are used for the manufacture of a medicament for treating osteoporosis by inhibiting the bone resorption activity of osteoclasts.

Description: BACKGROUND OF THE INVENTION This invention relates to a use for inhibiting bone resorption that is mediated by the action of a class of cells known as osteoclasts, involving compounds that compete with osteoclasts for the osteoclasts' site of activity. Osteoclasts are multinucleated cells of up to 400 µm in diameter that resorb mineralized tissue (chiefly, calcium carbonate and calcium phosphate) in vertebrates. They are actively motile cells that migrate along the surface of bone. They can bind to bone, secrete necessary acids and proteases and thereby cause the actual resorption of mineralized tissue from the bone. In the present invention, N-heterocyclicalkyl-substituted phenyl derivatives are used for the manufacture of a medicament for inhibiting the bone resorption activity of mammalian osteoclasts. These compounds have the general structural formula wherein n is an integer of from 0 to 6; Y is chosen from 0, SO₂, -CONH, -NHCO-, CH₂ or R¹ is an unsubstituted or substituted saturated mono-or polycyclic hetero-cyclic ring system having 1, 2 or 3 heteroatoms where said heteroatoms are chosen from O, N or S and wherein said substituents are independently chosen from R²; R² is C₁₋₆ alkyl, aryl C₀₋₆ alkyl, hydroxyC₀₋₆alkyl, C₁₋₄ alkyloxyC₀₋₆alkyl, carboxyC₀₋₆alkyl, oxo, halogen, CF₃, C₀₋₄alkylaminoC₀₋₆alkyl or C₀₋₄dialkylaminoC₀₋₆alkyl; R³ is hydrogen, C₁₋₆ alkyl, hydroxyC₀₋₄alkyl C₁₋₄ alkyloxyC₀₋₄alkyl carboxyC₀₋₄alkyl halogen or CF₃; R⁴ is hydrogen, C₁₋₆ alkyl, aryl C₀₋₄ alkyl, C₁₋₆ alkylcarbonyloxymethyl; and R⁵ is C 1-alkyl , aryl C₀₋₄alkyl or heterocyclylC₀₋₄alkyl and the pharmaceutically acceptable salts thereof. A preferred group of compounds used according to the present invention are those defined for the general structural formula shown below wherein: n is an integer from 0 to 6, Y is chosen from O, -NHCO-, -CONH- or CH₂; R¹ is an unsubstituted or substituted 5 or 6-membered heterocyclic ring system having 1, 2 or 3 heteroatoms where said heteroatoms are independently chosen from N or O and wherein said substituents are C₁₋₆ alkyl, aryl C₁₋₃ alkyl, hydroxy, C₁₋₄ alkoxy or oxo; R² and R³ are independently hydrogen, C₁₋₆ alkyl, arylC₀₋₄alkyl, hydroxyC₀₋₂alkyl, C₁₋₄ alkoxyC₀₋₂alkyl, carboxyC₀₋₃alkyl or oxo; R⁴ is H or C₁₋₄ alkyl, and the pharmaceutically acceptable salts thereof. A more preferred group of compounds of the present invention are those adjusted for the general structural formula shown below wherein: n is an integer of from 1 to 6; Y is chosen from: O, -NHCO-, -CONH- or CH₂; R¹ is an unsubstituted or substituted 5 or 6-membered heterocyclic ring having 1 or 2 heteroatoms where said heteroatom is N and wherein said substituents are C₁₋₃ alkyl, hydroxy or oxo; and R² is C₁₋₆ alkyl or aryl C₀₋₄alkyl, and the pharmaceutically acceptable salts thereof. The compounds are useful in the treatment of poultry and mammals, including man. The current major bone diseases of public concern are osteoporosis, hypercalcemia of malignancy, osteopenia due to bone metastases, periodontal disease, hyperparathyroidism, periarticular erosions in rheumatoid arthritis, Paget's disease, immobilization-induced osteopenia, and glucocorticoid treatment. All these conditions are characterized by bone loss, resulting from an imbalance between bone resorption (breakdown) and bone formation, which continues throughout life at the rate of about 14% per year on the average. However, 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 in vertebrae immediately following menopause, a condition which leads to increased fracture risk. There are currently 20 million people with detectable fractures of the vertebrae due to osteoporosis-in the U.S. In addition, there are 250,000 hip fractures per year attributed to osteoporosis, which are associated with a 12% mortality rate within the first two years and 30% of the patients require nursing home care after the fracture. All the conditions listed above would benefit from treatment with agents which inhibit bone resorption. SUMMERY OF THE INVENTION By this invention there is provided a use of compounds according to the invention for the manufacture of a medicament for the treatment of mammals suffering from a bone condition caused or mediated by increased bone resorption. The compounds according to the invention, including the pharmaceutically acceptable salts thereof, inhibit the activity of mammalian osteoclasts. DETAILED DESCRIPTION OF THE INVENTION The term "pharmaceutically acceptable salts" shall mean non-toxic salts of the compounds of this invention which are generally prepared by reacting the free base with a suitable organic or inorganic acid. Representative salts include the following salts: Acetate Benzenesulfonate Benzoate Bicarbonate Bisulfate Bitartrate Borate Bromide Calcium Edetate Camsylate Carbonate Chloride Clavulanate Citrate Dihydrochloride Edetate Edisylate Estolate Esylate Fumarate Gluceptate Gluconate Glutamate Glycollylarsanilate Hexylresorcinate Hydrabamine Hydrobromide Hydrochloride Hydroxynaphthoate Iodide Isothionate Lactate Lactobionate Laurate Malate Maleate Mandelate Mesylate Methylbromide Methylnitrate Methylsulfate Mucate Napsylate Nitrate Oleate Oxalate Pamaote Palmitate Pantothenate Phosphate/diphosphate Polygalacturonate Salicylate Stearate Subacetate Succinate Tannate Tartrate Teoclate Tosylate Triethiodide Valerate Additionally included are suitable cations such as alkali metal and alkaline earth cations. The term "pharmacologically effective amount" shall mean that amount of a drug or pharmaceutical agent that will elicit the biological or medical response of a tissue, system or animal that is being sought by a researcher or clinician. The term "bone resorption activity" shall mean the process by which osteoclasts solubilize bone minerals and increase the activity of enzymes that degrade bone matrix. The term "alkyl" shall mean straight or branched chain alkane, alkene or alkyne with one or more degrees of unsaturation at any position. In the schemes and examples below, various reagent symbols have the following meanings: The term "oxo" shall mean The term "heterocyclyl" shall mean a 5 or 6-membered mono or polycyclic ring system containing 1, 2, 3 or 4 heteroatoms chosen from O, N or S. The term "halogen" shall mean F, Cl, Br or I. The term "aryl" shall mean phenyl or benzyl. BOC: t-butoxycarbonyl Pd-C: Palladium on activated carbon catalyst. DMF: Dimethylformamide DMSO: Dimethylsulfoxide CBZ: Carbobenzyloxy EtOAc: ethyl acetate THF: tetrahydrofuran HOAc: acetic acid CHCl₃: chloroform MeOH: methanol CH₃CN: acetonitrile TFA: Trifluoroacetic acid BOP: Benzotriazol-4-yloxy-tris(dimethylamino)phosphonium hexafluorophosphate 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, elixers, tinctures, suspensions, syrups and emulsions. Likewise, they may also be administered in intravenous (bolus or infusion), intraperitoneal, subcutaneous or intramuscular form, all using forms well known to those of ordinary skill in the pharmaceutical arts. The dosage regimen utilizing the compounds of the present invention is selected in accordance with a variety of factors including type, species, age, weight, sex and medical condition of the patient; the severity of the condition to be treated; the route of administration; the renal and hepatic function of the patient; and the particular compound or salt thereof employed. An ordinarily skilled physician or veterinarian can readily determine and prescribe the effective amount of the drug required to prevent, counter or arrest the progress of the condition. Oral dosages of the present invention, when used for the indicated effects, will range between about 0.01 mg per kg of body weight per day (mg/kg/ day) to about 100 mg/kg/day and preferably 1.0-100 mg/kg/day and most preferably 1.0 to 20 mg/kg/day. Intravenously, the most preferred doses will range from about 1 to about 10 mg/kg/minute during a constant rate infusion. Advantageously, compounds of the present invention may be administered in a single daily dose, or the total daily dosage may be administered in divided doses of two, three or four times daily. Furthermore, preferred compounds for the present invention can be administered in intranasal form via topical use of suitable intranasal vehicles, or via transdermal routes, using those forms of transdermal skin patches well known to those of ordinary skill in that art. To be administered in the form of a transdermal delivery system, the dosage administration will, of course, be continuous rather than intermittant throughout the dosage regimen. In the use according to the present invention, the compounds herein described in detail can form the active ingredient, and are typically administered in admixture with suitable pharmaceutical diluents, excipients or carriers (collectively referred to herein as 'carrier' materials) suitably selected with respect to the intended form of administration, that is, oral tablets, capsules, elixirs, syrups and the like, and consistent with conventional pharmaceutical practices. For instance, for oral administration in the form of a tablet or capsule, the active drug component can be combined with an oral, non-toxic, pharmaceutically acceptable, inert carrier such as lactose, starch, sucrose, glucose, methyl cellulose, magnesium stearate, dicalcium phosphate, calcium sulfate, mannitol, sorbitol and the like; for oral administration in liquid form, the oral drug components can be combined with any oral, non-toxic, pharmaceutically acceptable inert carrier such as ethanol, glycerol, water and the like. Moreover, when desired or necessary, suitable binders, lubricants, 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. Lubricants used in these dosage forms include sodium oleate, sodium stearate, magnesium stearate, sodium benzoate, sodium acetate, sodium chloride and the like. Disintegrators.include, without limitation, starch, methyl cellulose, agar, bentonite, xanthan gum and the like. The compounds of the present invention can also be administered in the form of liposome delivery systems, such as small unilamellar vesicles, large unilamellar vesicles and multilamellar vesicles. Liposomes can be formed from a variety of phospholipids, such as cholesterol, stearylamine or phosphatidylcholines. Compounds of the present invention may also be delivered by the use of monoclonal antibodies as individual carriers to which the compound molecules are coupled. The compounds of the present invention may also be coupled with soluble polymers as targetable drug carriers. Such polymers can include polyvinylpyrrolidone, pyran copolymer, polyhydroxypropylmethacrylamidephenol, polyhydroxyethylaspartamidephenol, or polyethyleneoxide-polylysine substituted with palmitoyl residues. Furthermore, 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, polyglycolic acid, copolymers of polylactic and polyglycolic acid, polyepsilon caprolactone, polyhydroxy butyric acid, polyorthoesters, polyacetals, polydihydropyrans, polycyanoacrylates and crosslinked or amphipathic block copolymers of hydrogels. The compounds used in the method of the present invention were prepared according to the procedure of the following schemes and examples, using appropriate materials and are further exemplified by the following specific examples. The most preferred compounds are any or all of those specifically set forth in these examples. These compounds are not, however, to be construed as forming the only genus that is considered as the invention, and any combination of the compounds or their moieties may itself form a genus. The following examples further illustrate details for the preparation of the compounds of the present invention. Those skilled in the art will readily understand that known variations of the conditions and processes of the following preparative procedures can be used to prepare these compounds. All temperatures are degrees Celsius unless otherwise noted. EXAMPLE 1 This example is for illustrative purposes, as it is cross-referenced in subsequent examples describing preparation of compounds according to the invention. 2-S-(N-Benzyloxycarbonylamino)-3-[4-(3-t-butyl-aminopropyloxy)phenyl]propionic acid (1-2) Compound 1-1, 0.4 g, 1.1 mmole, was refluxed in t-butylamine (20 ml) and acetonitrile (20 mL) for three days. The reaction was evaporated to dryness, the residue dissolved in water, and extracted with ether. The aqueous layer was then acidified to pH 4-5 and a precipitate formed. The solid was collected and dried to yield 1-2. Rf = 0.8 in 9:1 EtOH/NH₄OH, ninhydrin stain. 300 MHz ¹H NMR (D₂O + NaOH) δ 7.4 (bs, 2H), 7.2 (bs, 4H), 6.85 (d, J = 8.55, 2H), 5.2 (d, J = 12.8 Hz, 1H), 5.0 (d, J = 12.8 Hz, 1H), 4.3 (dd, J = 4.0, 9.6 Hz, 1H), 4.0 (bs, 2H), 3.2(dd, J = 4.0, 13.6 Hz, 1H), 2.8 (dd, J = 9.6 Hz, 13.6 Hz, 1H), 2.65 (t, J = 7.3 Hz, 2H), 1.8 (m, 2H), 1.09 (s, 9H), mass spec (FAB) m/e = 429 (m + 1) C, H, N analysis C₂₄H₃₂N₂O₅ 0.65 H₂O Calculated C = 65.47, H = 7.62, N = 6.36 Found C = 65.52, H = 7.54, N = 6.27 MW = 440.244 EXAMPLE 2 This example is for illustrative purposes, as it is cross-referenced in subsequent examples describing preparation of compounds according to the invention. 2-S-(N-Benzyloxycarbonylamino)-3-[4-(3-chloro-propyloxy)phenyl]propionic acid (1-1) N-CBZ-tyrosine (3 g, 9.9 mmole) (Bachem Chemical Supply, California), was dissolved in DMF and treated with NaH (50% dispersion in oil, 0.95 g, 19.8 mmole) for 1 hour. Then 1,3-bromochloropropane (1 ml, 9.9 mmole) was added and the reaction stirred for 16 hours. The DMF was removed in vacuo and the residue dissolved in water, acidified to pH 3, and extracted with ethyl acetate. The ethyl acetate layer was dried with MgSO₄, filtered and evaporated. Column chromatography (SiO₂, 97:3:1 CHCl₃/CH₃OH/HOAc) yielded 1-1 as a yellow oil. R f = 0.3 in 97:3:1 CHCl₃/CH₃OH/HOAc ninhydrin stain ¹H NMR (300MHz, CDCl₃) δ 7.3 (bs, 5H), 7.03 (d, J = 8.3, 2H), 6.8 (d, J = 8.3, 2H), 5.2 (d, J = 8Hz, 1H), 5.05 (bs, 2H) 4.65 (m, 1H), 4.05 (t, J = 5.7 Hz, 2H), 3.73 (t, J = 6.3 Hz, 2H), 3.1 (m, 2H), 2.2 (m, 2H). EXAMPLE 3 2-S-(N-Benzyloxycarbonylamino)-3-[4-(3-N-pyrolidinyl-propyloxy)phenyl]propionic acid (1-4) Treatment of compound 1-1 with pyrrolidine in CH₃CN at reflux for three days provided crude 1-4. This was purified by flash chromatography on silica gel eluting with 9:1:1 EtOH/H₂O/NH₄OH to give pure 1-4 (R f = 0.81, ninhydrin stain). ¹H NMR (300 MHz. CDCl₃) δ 7.3 (bs, 5H), 7.0 (d, J = 8.1Hz, 2H), 6.7 (d, J = 8.1Hz, 2H), 5.5 (d, J = 7.4Hz, 1H), 5.0 (bs, 2H), 4.5 (m, 1H), 3.8 (m, 2H), 3.75 (bs, 1H), 3.4 (m, 2H), 3.18 (t, J = 8.5Hz, 2H), 3.1 (bs, 2H), 2.8 (bs, 1H), 2.2-1.8 (m, 6H). C, H, N analysis C₂₄H₃₀N₂O₅·0.25CH₂Cl₂ Calcd: C, 65.05; H, 6.87; N, 6.26. Found: C, 65.28; H, 6.78; N, 6.27. EXAMPLE 4 2-S-(N-Benzyloxycarbonylamino)-3-[4-(4-piperazinyl)-butyloxyphenyl]propionic acid (1-7) Treatment of N-CBZ-L-tyrosine with sodium hydride in DMF followed by 1,4-dibromobutane, as described for the preparation of 1-1, provided the corresponding butyl analog. Treatment of this with 1,4-piperazine in refluxing acetonitrile for three days gave crude 1-7 as a precipitate from the reaction mixture. Reverse phase HPLC purification gave pure 1-7. ¹H NMR (300 MHz, CD₃OD) δ 7.3 (m, 5H), 7.23 (d, 2H), 6.83 (d, 2H), 5.0 (bs, 2H), 4.35 (dd, 1H), 4.0 (t, 2H), 3.6 (bs, 8H), 3.1 (dd, 1H), 2.85 (dd, 1H), 2.00-1.8 (m, 4H). C, H, N analysis C₂₆H₃₅N₃O₅·1.2H₂O Calcd: C, 63.57; H, 7.67; N, 8.56. Found: C, 63.33; H, 7.28; N, 8.55. MW = 491.206 EXAMPLE 5 2-S-(N-Benzyloxycarbonyl)-3-[4-(4-methylpiperazin-1-yl)-propyloxyphenyl]propanoic acid (1-9) Treatment of 1-1 with N-methylpiperazine as described for 1-2 gave crude 1-9. This was purified by column chromatography on silica gel eluting with 9:1:1 C₂H₅OH/H₂O/NH₄OH to give pure 1-9 as the TFA salt. ¹H NMR (300 MHz, D₂O) δ 7.5 (3H, m), 7.4 (2H, d), 7.0 (2H, d), 5.18 (1H, d), 5.05 (1H, d), 4.5 (1H, m), 4.2 (2H, t), 3.8 (8H, s), 3.6 (2H, t), 3.3 (1H, m), 3.1 (3H, s), 3.0 (1H, m), 2.4 (2H, m). Analysis for C₂₅H₃₃N₃O₅·2.3CF₃CO₂H Calcd: C, 49.52; H, 4.96; N, 5.85. Found: C, 49.42; H, 4.98; N, 6.01. EXAMPLE 6 2-S-(N-Benzyloxycarbonylamino)-3-[4-(4-piperazin-1-yl)pentyloxyphenyl]propionic acid (1-11) 1-10 (0.658 g, 1.42 mmole), was dissolved in 30 mL CH₃CN and 1,4-piperazine (1.22 g, 14.16 mmole) was added. This solution was stirred at room temperature for 4 days. The solvent was then removed and the residue was dry packed on a silica gel column and eluted with 18:1:1 C₂H₅OH/H₂O/NH₄OH to give pure 1-11 as a white solid. ¹H NMR (300 MHz, CD₃OD) δ 1.52 (4H, m), 1.77 (2H, m), 2.40 (2H, t), 2.59 (4H, m), 2.80-2.94 (1H, m), 3.01-3.12 (5H, m), 3.94 (2H, m), 4.21 (1H, m), 6.76 (2H, d), 7.09 (2H, d). Analysis for C₂₆H₃₅N₃O₅·1.2 H₂O Calcd: C, 63.57; H, 7.67; N, 8.56 Found: C, 63.33; H, 7.28; N, 8.55 EXAMPLE 7 This example is for illustrative purposes, as it is cross-referenced in subsequent examples describing preparation of compounds according to the invention. 2-S-(N-Benzyloxycarbonylamino)-3-[4-(6-N-t-butyloxy-carbonylaminohexyloxy)phenyl]propionic acid (2-1) N-CBZ-L-tyrosine (15.0 g, 0.045 moles) was dissolved in 75 mL DMF and added at 0-10°C to a suspension of sodium hydride (2.16 g, 0.09 moles) in (-SOUTH-). The resulting suspension was stirred at 0-10°C for 1.0 hour and then 6-(N-t-butyloxycarbonylamino)hexyl bromide (12.6 g, 0.045 moles) in 25 mL DMF was added dropwise at 0-5°C and the clear, dark reaction mixture was stirred at room temperature overnight. After solvent removal, the residue was taken up in EtOAc and this was made acidic with 10% KHSO₄ solution. The organic phase was separated, washed with brine, dried (Na₂SO₄) and the solvent removed to give an oil. This was purified by column chromatography on silica gel eluting with 98:2:1 CHCl₃/CH₃OH/HOAc to give pure 2-1 as a clear oil. ¹H NMR (300 MHz CD₃OD) δ 1.45 (15H, m), 1.75 (2H, m), 2.80-3.15 (6H, m), 3.91 (2H, t), 4.38 (1H, m), 4.95 (6H, m), 6.79 (2H, d), 7.10 (2H, d), 7.28 (5H, m). EXAMPLE 8 This example is for illustrative purposes, as it is cross-referenced in subsequent examples describing preparation of compounds according to the invention. 2-S-(N-Benzyloxycarbonylamino)-3-[4-(6-aminohexyloxy-phenyl)]propionic acid hydrochloride (2-2) Compound 2-1 (51.4 mg, 0.1 mmole) was dissolved in 20 mL EtOAc and cooled to -20°C under N₂. HCl gas was bubbled into this solution for 10 minutes as the temperature rose to -5°C. The reaction mixture was stoppered and stirred at 0 to -5°C for 1 hour. The solvent was then removed in the rotary evaporator and the residue was triturated with ether to give 2-2 as a white solid. Rf = 0.4 (SiO₂, 9:1:1 EtOH/NH₄OH, H₂O). ¹H NMR (300 MHz, CD₃OD) δ 1.45 (6H, m), 1.73 (4H, m), 2.90 (3H, m), 3.13 (1H, m), 3.95 (2H, m), 4.30 (1H, bs), 6.77 (2H, d), 7.10 (2H, d), 7.32 (4H, m). Analysis for C₂₃H₃₁N₂O₅Cl·0.5 H₂O Calcd: C, 60.05; H, 7.01; N, 6.09 Found: C, 60.08; H, 7.06; N, 6.09 EXAMPLE 9 This example is for illustrative purposes, as it is cross-referenced in subsequent examples describing preparation of compounds according to the invention. 2-S-(N-Benzyloxycarbonylamino)-3-[4-(8-N-t-butyloxy-carbonylaminooctyloxy)phenyl]propionic acid (2-5) N-CBZ-L-tyrosine·H₂O (1.5 g, 4.29 mmole) was dissolved in EtOAc/CH₂Cl₂, dried over MgSO₄, filtered and evaporated. The residue was dissolved in DMF and treated with NaH (50% dispersion in oil, 0.43 g, 8.96 mmole) for 1 hour. N-BOC-8-amino-1-bromooctane (1.33 g, 4.34 mmole) was added and the reaction was stirred for 16 hours. The DMF was removed in vacuo , the residue dissolved in water, acidified to pH 3 and extracted with EtOAc. The EtOAc layers were combined, dried and concentrated. Column chromatography (SiO₂, 97:3:1 CHCl₃/MeOH/HOAc) gave 2-5. ¹H NMR (300 MHz, CD₃OD) δ 7.3 (m, 5H), 7.1 (d, 2H), 6.78 (d, 2H), 5.0 (2q, 2H), 4.38 dd, 1H), 3.8 (m, 2H), 3.13 (dd, 1H), 3.0 (t, 2H), 2.85 (dd, 1H), 1.75 (m, 2H), 1.4 (s, 9H), 1.35 (m, 3H), 1.3 (bs, 7H). EXAMPLE 10 2-(4-N-t-Butyloxycarbonylpiperidin-4-yl)ethanol (2-10) 4-piperidine-2-ethanol (Available from Aldrich) (130 g, 1.0 mole) was dissolved in 700 mL dioxane, cooled to 0°C and treated with 3 N NaOH (336 mL, 1.0 mole), and di-t-butylcarbonate (221.8 g, 1.0 mole). The ice bath was removed and the reaction stirred overnight. The reaction was concentrated, diluted with water and extracted with ether. The ether layers were combined, washed with brine, dried over MgSO₄, filtered and evaporated to give 2-10. R f = 0.37 in 1:1 EtOAc/Hexanes, ninhydrin stain ¹H NMR (300 MHz, CDCl₃) δ 4.07 (bs, 2H), 3.7 (bs, 2H), 2.7 (t, J = 12.5 Hz, 2H), 1.8-1.6 (m, 6H), 1.51 (s, 9H), 1.1 (ddd, J = 4.3, 12.5, 12 Hz, 2H). EXAMPLE 11 Methyl 4-(4-N-t-Butyloxycarbonylpiperidin-4-yl) but-2-enoate (2-11) Oxalyl chloride (55.8 mL, 0.64 mole) was dissolved in 1 L CH₂Cl₂ and cooled to -78°C under N₂. DMSO (54.2 mL, 0.76 mole) was added dropwise. After gas evolution had ceased, 2-10 (102.5 g, 0.45 mole) dissolved in 200 mL CH₂Cl₂ was added over 20 minutes. After stirring an additional 20 minutes, triethylamine (213 mL, 1.53 mole) was added dropwise and the cold bath removed. After 1 and 1/2 hours TLC showed starting material gone. Carbomethoxytriphenylphosphorane (179 g, 0.536 mole) was added and the reaction stirred overnight. The solution was diluted with 300 mL Et₂O, extracted once with 800 mL H₂O, twice with 300 mL 10% KHSO₄ solution, then once with 300 mL brine. The organic layer was dried over MgSO₄, filtered and evaporated. Column chromatography (SiO₂, 5% EtOAc/Hexanes) yielded of pure 2-11. ¹H NMR (300 MHz, CDCl₃) δ 6.9 (ddd J = 15.6, 7,6, 7.6 Hz, 1H), 5.8 (d, J = 15.6 Hz, 1H), 4.0 (bs, 2H), 3.7 (s, 3H), 2.6 (t, J = 12.6 Hz, 2H, 2.1 (t, J = 7.4 Hz, 2H), 1.7-1.4 (m, 3H), 1.4 (s, 9H), 1.1 (m, 2H). EXAMPLE 12 4-(4-N-t-Butyloxycarbonylpiperidin-4-yl)-butyl bromide (2-12) Compound 2-11 (36.2 g, 0.128 mole), was dissolved in 500 mL EtOAc. 10% Palladium on carbon (10 g) was added as a slurry in EtOAc and the reaction was placed under H₂ (in a balloon) overnight. The reaction was filtered through Solka-Floc, the cake washed with EtOAc and the ethyl acetate evaporated to give 4-(4-N-t-butyloxycarbonylpiperidin-4-yl)-butanoate. TLC R f = 0.69 in 30% EtOAc/Hexanes. ¹H NMR (300 MHz, CDCl₃) δ 4.0 (bs, 2H), 3.6 (s, 3H), 2.60 (t, J = 12.3 Hz, 2H), 2.20 (t, J = 7.4, 2H), 1.6 (m, 4H), 1.40 (s, 9H), 1.40 (m, 1H), 1.20 (m, 2H), 1.0 (m, 2H). The butanoate ester (45.3 g, 0.159 mole) was dissolved in CH₃OH and treated with 1 N NaOH (500 mL, 0.5 mole) overnight. The solvent was removed in vacuo, water was added and the solution washed with ether, then acidified with 10% KHSO₄ solution. The aqueous layer was washed with ether, the ether layers were combined, washed with brine and dried over MgSO₄ and concentrated to give the corresponding acid as a clear oil. ¹H NMR (300 MHz, CDCl₃) δ 4.0 (bs, 2H), 2.6 (m, 2H), 2.25 (m, 2H), 1.6 (bs, 4H, 1.4 (s, 9H), 1.3-0.9 (9H). This acid (20.4 g, 0.077 moles) was treated with borane (BH₃/THF, 235 mL, 235 mmole) in THF at 0°C for 1 hour. NaOH (1N, 250 mL) was added dropwise and the solution stirred overnight. The reaction was concentrated to remove THF, extracted with ether, the ether extracts were combined, dried over MgSO₄, filtered and evaporated to give the corresponding alcohol as a colorless oil. R f = 0.7 in 2:1 ethyl acetate/hexanes. ¹H NMR (300 MHz, CDCl₃) δ 4.1 (bs, 2H), 3.6 (t, 2H), 2.65 (t, 2H), 2.1 (bs, 1H), 1.65 (bs, 2H), 1.55 (m, 2H), 1.4 (s, 9H), 1.35 (m, 3H), 1.25 (m, 2H), 1.1 (m, 2H). This alcohol (19.7 g, 76.5 mmole) was dissolved in THF and treated with triphenylphosphine (23.1 g, 88 mmol) and cooled to 0°C. Carbon tetrabromide (29.8 g, 89.9 mmol) was added in one portion, the cold bath was removed and the reaction stirred overnight. Additional triphenylphosphine (11.71 g) and carbon tetrabromide (14.9 g) was added to drive the reaction to completion. The mixture was filtered and the liquid was diluted with ether and filtered again. After solvent removal the resulting liquid was adsorbed onto SiO₂ and chromatographed with 5% EtOAc/Hexanes to yield 2-12 as a clear colorless oil. R f = 0.6 in 1:4 ethyl acetate/hexanes ¹H NMR (300 MHz, CDCl₃) δ 4.1 (bs, 2H), 3.4 (t, 2H), 2.65 (t, 2H), 1.85 (m, 2H), 1.65 (bd, 2H), 1.4 (s, 9H), 1.35 (m, 2H), 1.3 (m, 3H), 1.1 (m, 2H). EXAMPLE 13 2-S-(N-Benzyloxycarbonylamino)-3-[4-(4-N-t-butyloxycarbonylpiperidin-4-ylbutyloxy) phenyl]propionic acid (2-13) N-CBZ-L-tyrosine was alkylated with 2-12 as taught for compounds 2-5 in Example 9 to provide 2-13. R f = 0.15 in 97:3:1 CHCl₃/CH₃OH/HOAc, iodine stain. ¹H NMR (300 MHz, CDCl₃) δ 7.2 (d, J = 7.5 Hz, 2H), 7.1 (d, J = 7.5 Hz, 2H), 7.0 (d, J = 7.3 Hz, 2H), 6.8 (d, J = 7.3 Hz, 2H), 5.2 (d, J = 7.9 Hz, 1H), 5.1 (s, 2H), 4.6 (m, 1H), 4.01 (bd, 2H), 3.92 (t, J = 6 Hz, 2H), 3.7 (m, 2H), 2.65 (bt, 7H), 1.75-1.4 (m, 7H), 1.45 (s, 9H), 1.3 (m, 2H), 1.1 (m, 2H). EXAMPLE 14 2-S-(N-Benzyloxycarbonylamino)-3-[4-(4-piperidin-4-ylbutyloxy)phenyl]propionic acid (2-14) Compound 2-13 was deprotected as taught for compound 2-2 in Example 8. The solvent was removed on the rotary evaporator and the residue was dissolved in water and extracted with ethyl acetate. The water layer was concentrated to dryness, evaporated and the residue was chromatographed (SiO₂, 9:1:1 EtOH/H₂O/NH₄OH). A small portion was then purified further by HPLC and isolated as the TFA salt. ¹H NMR (300 MHz, CD₃OD) δ 7.3 (m, 5H), 7.1 (d, 2H), 6.8 (d, 2H), 5.0 (q, 2H), 2.93 (t, 2H), 2.85 (dd, 1H), 1.92 (bd, 2H), 1.75 (m, 2H), 1.6-1.45 (m, 3H), 1.35 (m, 4H). Mass Spec. (FAB) m/e = 455 (m + 1). 2-S-Amino-3-[4-(4-N-t-butyloxycarbonyl-piperidin-4-yl)butyloxyphenyl]propionic acid (5-1) 2-13 (2.0 g) was dissolved in 100 mL EtOH, and 0.2 g 10% Pd/C was charged. This suspension was hydrogenated at balloon pressure overnight. Solvent removal provided 5-1 as a white solid. ¹H NMR (300 MHz, CD₃OD), δ 0.97-1.12 (2H, m), 1.20-1.54 (14H, m), 1.72 (4H, m), 2.71 (2H, m), 2.90-3.00 (1H, m), 3.22 (1H, dd), 3.30 (1H, m), 3.71 (1H, m), 3.95-4.10 (4H, m), 6.88 (2H, d), 7.21 (2H, d). EXAMPLE 16 2-S-(Pentanoylamino)-3-[4-(4-N-t-butyloxy-carbonylpiperidin-4-yl)butyloxyphenyl] propanoic acid (5-2) 5-1 (1.05 g, 2.5 mmole) was added to a cold solution of 1 N NaOH (2.5 mL) in 20 mL H₂O and stirred at 0-10° for 5 minutes to give a clear solution. Then, pentanoyl chloride (0.332 g, 2.75 mmole) was added dropwise followed by NaHCO₃ (0.231 g, 2.75 mmole) and the resulting mixture was stirred vigorously at 0-10° for 1 hour. The reaction mixture was diluted with H₂O (75 mL), acidified to pH 2-3 with 10% KHSO₄ and extracted with EtOAc. This extract was filtered, washed with brine, dried (Na₂SO₄) and the solvent removed to give an oil. This was purified by flash chromatography on silica gel eluting with 97:3:1 CHCl₃/CH₃OH/HOAc to give pure 5-2 as a clear oil. ¹H NMR (300 MHz, CD₃OD) δ 0.90 (3H, t), 1.20-1.62 (16H, m), 1.72 (2H, m), 2.14 (2H, m), 2.30 (8H, m), 2.65-2.90 (4H, m), 3.30 (1H, m), 3.93 (2H, m), 4.61 (1H, m), 6.81 (2H, d), 7.12 (2H, d). EXAMPLE 17 2-S-(Pentanoylamino)-3-[4-(4-piperidinylbutyloxy)phenyl]propionic acid hydrochloride (5-3) 5-2 (0.449 g), was dissolved in 30 mL EtOAc and treated with HCl gas at -10° as described for 2-2. The resulting solid was triturated with 40 mL Et₂O to give pure 5-3 as a white solid. ¹H NMR (300 MHz, CD₃OD) δ 0.85 (3H, t), 1.19 (2H, m), 1.30-1.65 (9H, m), 1.73 (2H, m), 1.95 (2H, m), 2.15 (2H, m), 2.80-3.02 (3H, m), 3.14 (1H, dd), 3.30-3.40 (3H, m), 3.95 (2H, t), 4.61 (1H, m), 6.82 (2H, d), 7.13 (2H, d). Analysis for C₂₃H₃₆N₂O₄·HCl·0.75 H₂O Calc.: C = 60.77, H = 8.54, N = 6.16 Found: C = 60.97, H = 8.39, N = 6.06. EXAMPLE 18 2-S-(Hexanoylamino)-3-[4-(4-N-t-butyloxy-carbonylpiperidin-4-yl)butyloxyphenyl]-propionic acid (5-4) 5-1 (0.41 g) was treated with hexanoyl chloride (0.21 mL, 1.50 mmole) as described for 5-2. Crude product was purified by flash chromatography on silica gel eluting with 97:3:1 CHCl₃/CH₃OH/HOAc to give pure 5-4. ¹H NMR (300 MHz, CD₃OD) δ 0.85 (3H, t), 0.97-1.35 (8H, M), 1.37-1.53 (12H, m), 1.60-1.80 (4H, m), 2.13 (2H, t), 2.80 (2H, m), 2.85 (1H, m), 3.12 (1H, dd) 3.90 (2H, t), 4.04 (2H, d), 4.62 (1H, m), 6.80 (2H, d), 7.12 (2H, d). EXAMPLE 19 2-S-(Hexanoylamino)-3-[4-(4-piperidin-4-yl-butyloxy)phenyl]propionic acid (5-5) 5-4 (0.199 g) was dissolved in 25 mL EtOAc and treated with HCl gas as described for compound 2-2 to provide pure 5-5. ¹H NMR (300 MHz, CD₃OD) δ 0.84 (3H, t), 1.08-1.20 (4H, m), 1.35 (4H, m), 1.52 (4H, m), 1.77 (2H, m), 1.92 (2H, d), 2.16 (2H, t), 2.80-3.-2 (3H, m), 3.15 (1H, dd), 3.40-3.52 (2H, m), 3.92 (2H, t), 4.61 (1H, m), 6.81 (2H, d), 7.13 (2H, d). Analysis for C₂₆H₃₉N₂O₆F₃·0.55 H₂O·0.30 TFA Calc.: C = 55.39, H = 7.06, N = 4.86 Found: C = 55.38, H = 7.03, N = 4.85. Sample alternative protecting groups that can be used in the preparation of the present invention include benzyl ester, cyclohexyl ester, 4-nitrobenzyl ester, t-butyl ester, 4-pyridylmethyl ester, benzyloxycarbonyl, isonicotinyloxycarbonyl, O-chlorobenzyloxycarbonyl, p-nitrobenzyloxycarbonyl, p-methoxybenzyloxycarbonyl, t-butoxycarbonyl, t-amyloxycarbonyl, isobornyloxycarbonyl, adamantyloxycarbonyl, 2-(4-biphenyl)-2-propyloxycarbonyl and 9-fluorenylmethoxycarbonyl. In addition to those compounds specifically exemplified above, additional compounds of the present invention are set forth in tabular form below. These compounds are synthesized by use of the synthetic routes and methods described in the above Schemes and Examples and variations thereof well known to those of ordinary skill in the art, and not requiring undue experimentation. All variables listed in the Tables below are with reference to the following generic structure: The test procedures employed to measure the osteoclast inhibiting activity of the compounds used in the method of the present invention are described below. EXAMPLE 20 "When osteoclasts engage in bone resorption, they will literally 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 cross sections (4.4 x 4.4 x 0.2 mm) of bovine femur were cut from the diaphysis with a low-speed diamond saw (Isomet, Buehler, Ltd., Lake Bluff, IL) by the method of Arnett and Dempter. Endocrinology 120:602-608. Prior to incubation with osteoclasts, slices were rehydrated in 0.1 ml complete medium 199 in a 48-well plate (Costar, Cambridge, MA) overnight in the presence of twice the desired dose of compound being tested. Osteoclasts were isolated from the long bones of 1 to 3-day-old rats (Sprague-Dawley) by adaptations of methods used by Chambers, et al. J. Cell Sci. 66:383-399. Femora, tibiae, and humeri were split and minced with scalpel blades into 2-5 ml Medium 199 (GIBCO, New York). The resulting suspension was gently pipetted (60 times with a wide-bore pipet and then aliquoted onto petri dishes (Costar) or bone slices (0.1 ml per slice). Cells were allowed to settle for 30-40 minutes at 37°C in moist CO₂-air before gentle washing and reincubation in undiluted incubation medium. Osteoclast yields varied from 300 to 1400 per rat and typically comprised 1% or less of the total cell population. Osteoclasts were counted at the day of isolation and after 1 day of incubation by phase-constrast microscopy (Nikon Diaphot). Total attached cells were counted 50-70 h after isolation with a Coulter counter (model ZM, Coulter Electronics, Inc., Hialeah, FL). Cell counts of controls varied from 3.352 x 10⁴ to 2.322 x 10⁵ per well. Counting mononuclear cells at the time of isolation was not practical because of matrix and cell debris that could not be completly eliminated. Bone slices exposed to osteoclasts for 20 h after isolation were processed for staining by ultrasonication (twofold, 15 s, Branson) in 0.25 M ammonium hydroxide before fixation (20 minutes) in 2.5% glutaraldehyde, 0.1 M cacodylate, pH 7.4 (EM Supplies, Fort Washington, PA). Samples were dehydrated in ethanol (40, 70, and 100%; 5 minutes), air dried for 2 h, and then stained for 4 minutes with filtered 1% toluidine blue and 1% borax (Sigma, St. Louis, MO). Samples used to count osteoclasts were processed as earlier without ultrasonication in ammonium hydroxide. Samples processed for scanning electron microscopy were not air dried but infiltrated for 40 minutes with 1:1 ethanol-Peldri II (Ted Pella, Inc., Redding, CA). After incubation in 100% Peldri II, solidified samples were exacuated overnight to sublimate the Peldri II. Slices were rotary shadowed with gold (DV-502A, Denton Vacuum, Cherry Hill, NJ) and then examined on a JEOL JSM 840 at 3 kV accelerating voltage. The morphology and motility of living osteoclasts were analyzed by recording phase-contrast images (Nikon, NY) in real time onto 3/4 inch video-tapes with a u-matic VCR (Model VO 5800H, Sony). A fluorescence microscope (Microphot, Nikon) was adapted for reflected light microscopy by inserting a λ/4 plate between cross polarizers in the epi mode. Fluorescence objectives of long working distance with adjustable correction collars (10x, 20x, Nikon) were fitted with rotatable λ/4 plates (Polaroid Corp., MA) mounted as the front element. Correction collars were necessary 20x objectives and higher to correct for the presence of the λ/4 plate and the absence of a coverslip. Coverslips were not used to eliminate stray reflections below the λ/4 plate. Immersion oil (Nikon) was added between the objective front lens and λ/4 plate to minimize reflections at this interface. Oil was not placed between objective and specimen. Bone slices were scanned for resorption pits by rotating the λ/4 plate 0-45° with respect to the plane of polarization in epi-tungsten illumination. Alternatively, Hg illumination (HBO 100w, Nikon) was used with the λ/4 plate fixed at 45° while intermittently viewing stained images by transmission bright-field microscopy with an NCB 10 filter (Nikon. Quantitation of resorbed areas of bone slices examined by bright-field, RLM, and SEM was achieved through digital image processing (Magiscan 2A, Joyce Loebl, New York) of video images (Newvicon or SIT, Dage-MTI, Inc. Michigan City, IN) fed through a NTSC/PAL digital standards converter (CEL P156, James Grunder and Assoc., Inc., Mission, KS). Osteoclasts were processed for immunofluorescence by briefly rinsing coverslips in buffer S (60 mM Pipes, pH 6.9; 25mM Hepes; 10mM EGTA; and 2 mM MgCl₂) at 37°C and then fixing for 2 minutes in buffer S + 10% formaldehyde, pH 7.0. Cells were permeabilized in buffer S + 0.5% Triton X-100 and then rinsed. Specimens were incubated (30 minutes) in appropriate antibody or rhodamine-phalloidine (Molecular probes, Eugene, OR) followed by fluorescein goat antirabbit antibody (Cappel). The bone slice assay is used to examine the effect of the compound of interest on the activity of isolated osteoclasts from rat long bones. The number of resorption pits formed by osteoclasts after 1 day on consecutive cross sections of bovine femur was first compared to control samples by the method of Arnett and Dempster. Endocrinology 120:602-608, and then plotted as a function of concentration of the compound of interest. This is shown at Fig. 1 The appropriateness of extrapolating data from this assay to utility and use in 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. 1, 1990. That article teaches that certain bisphosphonates have been used clinically and appear to be effective in the treatment of Paget's disease, hypercalcemia of malignancy, osteolytic lesions produced by bone metastases, and bone loss due to immobilization or sex hormone deficiency. These same bisphosphonates are then tested in the resorption pit assay described above to confirm a correlation between their known utility and positive performance in the assay. While the invention has been described and illustrated in reference to certain preferred embodiments thereof, those skilled in the art will appreciate that various changes, modifications and substitutions can be made therein without departing from the spirit and scope of the invention. For example, effective dosages other than the preferred doses as set forth hereinabove may be applicable as a consequence of variations in the responsiveness of the mammal being treated for severity of bone disorders caused by resorption, or for other indications for the compounds of the invention indicated above. Likewise, the specific pharmacological responses observed may vary according to and depending upon the particular active compound selected or whether there are present pharmaceutical carriers, as well as the type of formulation and mode of administration employed, and such expected variations or differences in the results are contemplated in accordance with the objects and practices of the present invention. It is intended, therefore, that the invention be limited only by the scope of the claims which follow and that such claims be interpreted as broadly as is reasonable.

Claims: The use of a compound of formula wherein n is an integer of from 0 to 6; Y is O, SO₂, -CONH-, -NHCO-, CH₂, or R¹ is an unsubstituted or substituted saturated monoor polycyclic heterocyclic ring system having 1, 2 or 3 heteroatoms where said heteroatoms are chosen from O, N or S and wherein said substituents are independently chosen from R²; R² is C₁₋₆ alkyl, aryl C₀₋₆ alkyl, hydroxy C₀₋₆ alkyl, C₁₋₄ alkoxy C₀₋₆ alkyl, carboxy C₀₋₆ alkyl, oxo, halogen, CF₃, C₀₋₄ alkylamino C₀₋₆ alkyl or C₀₋₄ dialkylamino C₀₋₆ alkyl; R³ is hydrogen, C₁₋₆ alkyl, hydroxy C₀₋₄ alkyl, C₁₋₄ alkoxy C₀₋₄ alkyl, carboxy C₀₋₄ alkyl, halogen or CF₃; R4 is hydrogen, C₁₋₆ alkyl, aryl C₀₋₄ alkyl, or C₁₋₆ alkylcarbonyloxymethyl; and R⁵ is C₁₋₆alkyl, aryl C₀₋₄ alkyl, or heterocyclyl C₀₋₄ alkyl, or a pharmaceutically acceptable salt thereof, for the manufacture of a medicament for inhibiting the bone resorption activity of mammalian osteoclasts. The use of a compound of formula wherein n is an integer of from 1 to 6; Y is chosen from O, -NHCO-, -CONH- or CH₂; R¹ is an unsubstituted or substituted 5 or 6-membered heterocyclic ring having 1 or 2 heteroatoms where said heteroatoms are independently chosen from O and N and wherein said substituents are C₁₋₆ alkyl, aryl C₁₋₃ alkyl, hydroxy, C₁₋₄ alkoxy or oxo, R² and R³ are independently hydrogen, C₁₋₆ alkyl, aryl C₀₋₄ alkyl, hydroxy C₀₋₂ alkyl, C₁₋₄ alkoxy C₀₋₂ alkyl or carboxy C₀₋₃ alkyl; and R⁴ is H or C₁₋₄ alkyl, or a pharmaceutically acceptable salt thereof, for the manufacture of a medicament for inhibiting the bone resorption activity of mammalian osteoclasts. The use of a compound of formula wherein n is an integer of from 1-6; Y is chosen from O, -NHCO-, -CONH- or CH₂; R¹ is an unsubstituted or substituted 5 or 6-membered heterocyclic ring having 1 or 2 heteroatoms where said heteroatom is N and wherein said substituents are C₁₋₃ alkyl, hydroxy or oxo; and R² is C₁₋₆ alkyl, aryl C₀₋₄ alkyl for the manufacture of a medicament for inhibiting the bone resorption activity of mammalian osteoclasts. The use of a compound of formula wherein n is an integer of from 1 to 6; Y is an optional substituent that, when present, is O; R¹ is an unsubstituted or substituted 5 or 6-membered saturated heterocyclic ring having 1 or 2 heteroatoms where said heteroatom is N and said substituent is alkyl; and R² is alkyl, phenyl or phenalkyl, for the manufacture of a medicament for inhibiting the bone resorption activity of mammalian osteoclasts. The use of a compound of formula wherein n is an integer of from 1 to 4; Y is an optional substituent that, when present, is O; R¹ is unsubstituted or substituted 5 or 6-membered saturated heterocyclic rings having 1 or 2 heteroatoms where said heteroatom is N and said substituent is alkyl; and R² is alkyl of 1 to 4 carbon atoms or benzyl, for the manufacture of a medicament for inhibiting the bone resorption activity of mammalian osteoclasts. The use as claimed in Claim 2, in which said compound is The use as claimed in Claim 2, in which said compound is

Inventors: EGBERTSON MELISSA S, GOULD ROBERT J, HARTMAN GEORGE D, EGBERTSON, MELISSA S., GOULD, ROBERT J., HARTMAN, GEORGE D.
Applicants: MERCK & CO INC, MERCK & CO. INC.
Application year: 1992
Main classification: A61K31
IPC codes: A61K31, A61P19, A61P3, A61P43, C07C271, C07C317, C07D211