CA2334815A1 - Composition for treating cartilage disease - Google Patents

Abstract

This invention provides a pharmaceutical composition comprising a compound o f formula (I) wherein ring A is an optionally substituted benzene ring; R is a hydrogen atom or an optionally substituted hydrocarbon group; B is an optionally esterified or amidated carboxyl group; X is -CH(OH)- or -CO-; k i s 0 or 1; and k' is 0, 1 or 2, or its salt, which can be advantageously used a s an agent for prevention and/or treatment of a cartilage disease.

Description

DESCRIPTION
COMPOSITION FOR TREATING CARTILAGE DISEASE
Technical Field The present invention relates to a pharmaceutical composition for prevention and/or treatment of a cartilage disease, which comprise benzothiepine derivatives having excellent chondrogenesis promoting effect, cartilage destruction suppressing effect and cartilage cell differentiation induction promoting effect.
Background Art An articular disorder is a disease whose major lesion is a degeneration of an articular cartilage.
Cartilage is the organization composed by collagen and proteoglycan. Due to various causes, the synthesizing ability of proteoglycan in this cartilage organization declines , and proteoglycan starts to be released from the organization. The release of type-I collagenase (metalloprotease I) is simultaneously increased, and collagen of the cartilage organization is resolved. The destruction of the cartilage organization proceeds due to a series of these responses. And it undergoes, depending on the stage of the lesion, a hyperplasia of a synovial membrane, a destruction of a subcartilaginous bone, a hyperplasia or a neoplasia of a circumarticular cartilage, which are followed by a deformation of the cartilage, which may lead to dysfunction in a serious case. While the articular disorder occurs most frequently in a knee joint , it occurs also in the joints of elbows, thighs, legs and fingers . Among the articular diseases , the disease which is observed in the largest number of patients is an osteoarthritis, and it is considered to occur increasingly in an elderlies-dominating society in near future, since one of its causes is considered to be the aging of a human .
For treating this , an analgesic antiinflammatory agent or a hyaluronic acid formulation is employed to remedy the pain due to cartilage degeneration or subcartilaginous bone destruction. However, all therapeutic methods are only nosotropic, and exhibit no sufficient effects.
Suppression of cartilage destruction, promotion of chondrogenesis and induction of cartilage cell differentiation are considered to be effective in prevention and treatment of a cartilage disease.
In the field of therapeutic and prophylactic agents against cartilage diseases which are no more than nosotropic currently, a novel cartilage disease preventing and/or treating agent which is rather radical and excellent in terms of the characteristics required in a useful pharmaceutical (e. g., such as stability, absorption, bioavailability) is demanded.
Disclosure of Invention We made much effort to develop a pharmaceutical capable of exerting a direct effect on a cartilage cell to suppress a cartilage destruction and also capable of promoting cartilaginous osteoanagenesis, and finally discovered that Compound (I):
(CH2)k-~B ~ (I) A ~ S(=~~k X~R
wherein ring A is an optionally substituted benzene ring; R is a hydrogen atom or an optionally substituted hydrocarbon group; B is an optionally esterified or amidated carboxyl group; X is -CH(OH)- or -CO-; k is 0 or 1; and k' is 0, 1 or 2, and its salt exhibits excellent chondrogenesis promoting effect, cartilage destruction suppressing effect and cartilage cell differentiation induction promoting effect, and then we made a further effort based on these findings, whereby establishing the present invention.
Accordingly, the present invention relates to:
(1) a pharmaceutical composition for prevention and/or treatment of a cartilage disease, which comprises a compound (I) of the formula:
CCHz)k-~B
(I) X--~g wherein ring A is an optionally substituted benzene ring;
R is a hydrogen atom or an optionally substituted hydrocarbon group; B is an optionally esterified or amidated carboxyl group ; X is -CH ( OH ) - or -CO- ; k is 0 or 1; and k' is 0, 1 or 2, or its salt, (2) a pharmaceutical composition according to (1), wherein the ring A is a benzene ring which may be substituted by 1 or 2 substituents selected from the group consisting of a halogen, a C1_lo alkyl, a C1_lo alkoxy, an alkylenedioxy group of the formula : -O- ( CHZ } ~-O- wherein n is an integer from 1 to 3 and a Cl_lo alkylthio group; R is a hydrogen atom, a C1_6 alkyl or a phenyl group ; B is -CON ( Rl ) ( Rz ) wherein R1 is a hydrogen atom or a C1_lo alkyl group and R~ is a phenyl or a phenyl-C1_3 alkyl group which those groups may be substituted by a halogen, a Cl_6 alkoxy, a mono- or di-C1_6 alkoxyphosphoryl, a mono- or di-C1_6 alkoxyphosphoryl-C1_ 3 alkyl wherein two alkyl groups of these di-C1_6 alkoxy group may bind together to form a Cl_6 alkylene group and C1_6 alkoxycarbonyl group, (3) a pharmaceutical composition according to (1), wherein the compound is an optically active compound ( II ) of the formula;

/Og4 _.., ,~~cONA ~ ~ ~2P\ 5 _. ' C ;~ s a 0 g3 wherein R' is a Cl_6 alkyl group; and R' and RS are independently a C1_6 alkyl group or bind together to form a C1_6 alkylene group, (4) a pharmaceutical composition according to (3}, wherein R,, R~ and RS are independently a C1_4 alkyl group, (5) a pharmaceutical composition according to (3), wherein the optically active compound (II) is (2R,4S)-(-)-N-[4-(diethoxyphosphorylmethyl}phenyl)-1,2,4,5-tetrahydro-4-methyl-7,8-methylenedioxy-5-oxo-3-benzothiepine-2-carboxamide, (6) a pharmaceutical composition according to (1), which is in the form of a sustained-release preparation comprising a biodegradable polymer, (7) a pharmaceutical composition according to (1), which is for local administration, (8) a pharmaceutical composition according to (1), which is for oral administration, (9) a pharmaceutical composition according to (1), which is for injection, ( 10 ) a pharmaceutical composition according to ( 1 ) , which is for potent cartilage destruction suppressing agent, chondorogenesis promoting agent or cartilage cell differentiation induction promoting agent, (11) a pharmaceutical composition according to(1}, which is for proteoglycan synthesis promoting agent, type II collagen synthesis promoting agent, metalloprotease I
release suppressing agent or proteoglycan release suppressing agent, (12) a pharmaceutical composition according to (I), which is for proteoglycan synthesis promoting agent or metalloprotease I release suppressing agent, (13) a pharmaceutical composition according to (1), wherein the cartilage disease is a cartilage defect, 5 chronic rheumatoid arthritis and osteoarthritis, (14) a method for prevention and/or treatment of a cartilage disease, which comprises administrating Compound ( I ) or its salts defined in ( 1 ) to mammal in need, and (15) use of Compound (I) or its salts defined in ( 1 ) for manufacturing a medicament for prevention and/or treatment of a cartilage disease.
With respect to the Compound (I) or its salt, the substituent of the substituted benzene represented by ring A is exemplified by a halogen atom, a nitro group, an optionally substituted alkyl group, an optionally substituted hydroxyl group, an optionally substituted mercapto group, an optionally substituted amino group, an acyl group, a mono- or di-alkoxyphosphoryl group, a phosphono group, an optionally substituted aryl group, an optionally substituted aralkyl group and an optionally substituted aromatic heterocyclic group. Of these substituents , 1 to 4 , preferably 1 or 2 , whether identical or not, may be present on the benzene ring.
The halogen atom includes fluorine, chlorine, bromine and iodine.
The alkyl group of the optionally substituted alkyl group includes an alkyl group having 1 to 10 carbon atoms such as methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, tert-butyl, pentyl, isopentyl, neo-pentyl, hexyl, heptyl, octyl, nonyl and decyl, and a cycloalkyl group having 3 to 7 carbon atoms such as cyclopropyi, cyclobutyl, cyclohexyl and cycloheptyl. These alkyl groups may be substituted by 1 to 3 substituents selected from a halogen atom (e. g., fluorine, chlorine, bromine, iodine), a hydroxyl group, an alkoxy group having 1 to 6 carbon atoms (e. g., methoxy, ethoxy, propoxy, butoxy, hexyloxy) , a mono- or di-C1_6 alkoxyphosphoryl group ( a . g .
methoxyphosphoryl,ethoxyphosphoryl,dimethoxyphosphoryl, diethoxyphosphoryl) and a phosphono group.
The substituted alkyl group includes trifluoromethyl, trifluoroethyl, trichloromethyl, hydroxymethyl, 2-hydroxyethyl, methoxyethyl, 1-methoxyethyl, 2-methoxyethyl, 2,2-diethoxyethyl, 2-diethoxyphosphorylethyl, phosphonomethyl and so on.
The substituted hydroxyl group includes alkoxy group , an alkenyloxy group , an aralkyloxy group , an acyloxy group , an aryloxy group and so on . Preferable alkoxy groups is an alkoxy group having 1 to 10 carbon atoms ( e. g. , methoxy, ethoxy,propoxy,butoxy, tert-butoxy,pentyloxy, hexyloxy, heptyloxy, nonyloxy) and a cycloalkoxy group having 4 to 6 carbon atoms (e. g., cyclobutoxy, cyclopentoxy, cyclohexyloxy}. Preferable alkenyloxy group is an alkenyloxy group having 2 to 10 carbon atoms such as allyloxy, crotyloxy, 2-pentenyloxy, 3-hexenyloxy, 2-cyclopentenylmethoxy and 2-cyclohexenylmethoxy.
Preferable aralkyloxy group is an aralkyloxy group having 7 to 19 carbon atoms, with greater preference given to a Ce-a aryl-Ci_, alkyloxy group ( a . g . , benzyloxy, phenethyloxy). Preferable acyloxy group is an alkanoyloxy group such as one having 2 to 10 carbon atoms (e. g., acetyloxy, propionyloxy, n-butyryloxy, hexanoyloxy).
Preferable aryloxy group is aryloxy group having 6 to 14 carbon atoms (e.g., phenoxy, biphenyloxy). Further, these substituted hydroxy groups may be substituted by 1 to 3 substituents selected from the above-mentioned halogen atom, a hydroxyl group, an alkoxy group having 1 to 6 carbon atoms , a mono- or di-C1_6 alkoxyphosphoryl group, a phosphono group, etc. Specifically, the substituted hydroxyl group includes trifluoromethoxy, 2,2,2-trifluoroethoxy, difluoromethoxy, 2-methoxyethoxy, 4-chlorobenzyioxy and 2-(3,4-dimethoxyphenyl)ethoxy, and so on.

The substituted mercapto group includes an alkylthio group, an aralkylthio group and an acylthio group.
Preferable alkylthio group is an alkylthio group having 1 tolOcarbon atoms(e.g.,methylthio,ethylthio,propylthio, butylthio, pentylthio, hexylthio, heptylthio, nonylthio) and a cycloalkylthio group having 4 to 6 carbon atoms ( a . g . , cyclobutylthio, cyclopentylthio, cyclohexylthio).
Preferable aralkylthio group is an aralkylthio group having 7 to 19 carbon atoms , more preferably a C6_1~ aryl-C1_, alkylthio group such as benzylthio and phenethylthio.
Preferable acylthio group is alkanoylthio group such as one having 2 to 10 carbon atoms (e. g., acetylthio, propionylthio, n-butyrylthio, hexanoyithio). Further, these substituted mercapto groups may be substituted by 1 to 3 substituents selected from the above-mentioned halogen atom, a hydroxyl group, an alkoxy group having 1 to 6 carbon atoms , a mono- or di-Cl_6 alkoxyphosphoryl group , a phosphono group, etc. Specifically, the substituted mercapto group includes tri-fluoromethylthio, 2,2,2-trifluoroethylthio, 2-methoxyethylthio, 4-chlorobenzylthio, 3,4-dichlorobenzylthio, 4-fluorobenzylthio, 2-(3,4-dimethoxyphenyl)ethylthio, and so on.
As substituents of the substituted amino group, there may be used 1 or 2 of identical or different substituents selected from the above-mentioned alkyl group having 1 to 10 carbon atoms , an alkenyl group having 2 to 10 carbon atoms (e.g., allyl, vinyl, 2-penten-1-yl, 3-penten-1-yl, 2-hexen-1-yl, 3-hexen-1-yl, 2-cyclohexenyl, 2-cyclopentenyl, 2-methyl-2-propen-1-yl, 3-methyl-2-buten-1-yl ) , an aryl group having 6 to 14 carbon atoms ( a . g .
phenyl , naphthyl ) and an aralkyl group having 7 to 19 carbon atoms (e.g. benzyl, phenetyl). These substituents may be substituted by the above-mentioned halogen atom, an alkoxy group having 1 to 6 carbon atoms, a mono- or di-C1_6 alkoxyphosphoryl group, a phosphono group, etc.
Specifically, the substituted amino group includes methylamino, dimethylamino, ethylamino, diethylamino, dibutylamino,diallylamino,cyclohexylamino,phenylamino, N-methyl-N-phenylamino, N-methyl-N-(4-chlorobenzyl)amino and N,N-di(2-methoxyethyl)amino, and so on.
The acyl group includes an organic carboxylic acid acyl group and a sulfonic acid acyl group with a hydrocarbon group having 1 to 6 carbon atoms ( a . g . , Cl_6 alkyl group such as methyl, ethyl, n-propyl, hexyl) or phenyl. Useful organic carboxylic acyl group is formyl, a C1_lo alkyl-carbonyl group (e. g., acetyl, propionyl, butyryl, valeryl, pivaloyl, hexanoyl, octanoyl, cyclobutanecarbonyl, cyclohexanecarbonyl, cycloheptanecarbonyl), a Cz_lo alkenyl-carbonyl group (e.g., crotonyl, 2-cyclohexenecarbonyl), a C6_1, aryl-carbonyl group (e. g., benzoyl), a C,_19 aralkyl-carbonyl group (e. g., benzylcarbonyl, benzhydrylcarbonyl), a 5- or 6-membered aromatic heterocyclic carbonyl group (e. g, nicotinoyl, 4-thiazolylcarbonyl) or a 5- or 6-membered aromatic heterocyclic acetyl group (e.g., 3-pyridylacetyl, 4-thiazolylacetyl ) . Useful sulfonic acyl group having 1 to 6 carbon atoms are methanesulfonyl and ethanesulfonyl, and so on. These acyl groups may be substituted by 1 to 3 substituents selected from the above-mentioned halogen atom, a hydroxyl group, an alkoxy group having 1 to 6 carbon atoms, an amino group, etc. Specifically, the substituted acyl group includes trifluoroacetyl, trichloroacetyl, 4-methoxybutyryl, 3-cyclohexyloxypropionyl, 4-chlorobenzoyl and 3,4-dimethoxybenzoyl, and so on.
The mono- or di-alkoxyphosphoryl group includes a mono-C1_6 alkoxyphosphoryl group such as methoxyphosphoryl , ethoxyphosphoryl, propoxyphosphoryl, isopropoxyphosphoryl, butoxyphosphoryl, pentyloxyphosphoryl and hexyloxyphosphoryl, and a di-C1_6 alkoxyphosphoryl group such as dimethoxyphosphoryl, diethoxyphosphoryl, dipropoxyphosphoryl, diisopropoxyphosphoryl, dibutoxyphosphoryl, dipentyloxyphosphoryl and dihexyloxyphosphoryl, with preference given to a di-CI_6 alkoxyphosphoryl group such as dimethoxyphosphoryl, diethoxyphosphoryl, dipropoxyphosphoryl, diisopropoxyphosphoryl, ethylenedioxyphosphoryl, dibutoxyphosphoryl, etc.
The aryl group of the optionally substituted aryl group includes an aryl group having 6 to 14 carbon atoms such as phenyl, naphthyl and anthryl. These aryl groups may be substituted by 1 to 3 substituents selected from the above-mentioned alkyl group having 1 to 10 carbon atoms, a halogen atom, a hydroxyl group, an alkoxy group having 1 to 6 carbon atoms, etc. Specifically, the substituted aryl group includes 4-chlorophenyl, 3,4-dimethoxyphenyl, 4-cyclohexylphenyl and 5,6,7,8-tetrahydro-2-naphthyl.
The aralkyl group of the optionally substituted aralkyl group includes aralkyl group having 7 to 19 carbon atoms such as benzyl, naphthylethyl and trityl. These aralkyl groups may be substituted by 1 to 3 substituents selected from the above-mentioned alkyl group having 1 to 10 carbon atoms , a halogen atom, a hydroxyl group, an alkoxy group having 1 to 6 carbon atoms , etc . on the aromatic ring .
Specifically, the substituted aralkyl group includes 4-chlorobenzyl, 3,4-dimethoxybenzyl, 4-cyclohexylbenzyl and 5,6,7,8-tetrahydro-2-naphthylethyl.
The aromatic heterocyclic group of the optionally substituted aromatic heterocyclic group includes a 5- or 6-membered aromatic heterocyclic group having 1 to 4 atoms of nitrogen, oxygen and/or sulfur, such as furyl, thienyl, imidazolyl, thiazolyl, oxazolyl and thiadiazolyl. These aromatic heterocyclic groups may be substituted by 1 to 3 substituents selected from the above-mentioned alkyl group having 1 to 10 carbon atoms, a halogen atom, a hydroxyl group, an alkoxy group having 1 to 6 carbon atoms, etc.
Provided that two alkyl groups are present as mutually adjoining substituents on the benzene ring A, they may bind together to form an alkylene group represented by the formula : - ( CHZ ) m- wherein m is an integer from 3 to 5 (e. g., trimethylene, tetramethylene, pentamethylene).
Provided that two alkoxy groups are present as mutually 5 adjoining substituents on the benzene ring A, they may bind together to form an alkylenedioxy group represented by the formula: -O- ( CHZ ) "-O- wherein n is an integer from 1 to 3 (e.g.,methylenedioxy,ethylenedioxy,trimethylenedioxy).
In these cases, a 5- to ?-membered ring is formed in 10 cooperation with carbon atoms of the benzene ring.
With respect to the Compound (I), R is a hydrogen atom or an optionally substituted hydrocarbon group.
The hydrocarbon group of the optionally substituted hydrocarbon group represented by R is exemplified by the above-mentioned alkyl group (preferably an alkyl group having 1 to 10 carbon atoms such as methyl, ethyl, propyl, isopropyl,butyl,isobutyl,sec-butyl,tert-butyl,pentyl, iso-pentyl, neo-pentyl and hexyl), an alkenyl group (preferably an alkenyl group having 2 to 10 carbon atoms) , an aryl group (preferably an aryl group having 6 to 14 carbon atoms) and an aralkyl group (preferably an aralkyl group having ? to 19 carbon atoms ) . Useful substituents on the hydrocarbon group include the above-mentioned 5- or 6-membered aromatic heterocyclic groupsuch as furyl,thienyl, imidazolyl, thiazolyl, oxazolyl and thiadiazolyl , a halogen atom, a di-C1_6 alkoxyphosphoryl group and a phosphono group.
With respect to the Compound ( I ) , B is an optionally esterified or amidated carboxyl group.
The esterified carboxyl group represented by B is exemplified by an alkoxycarbonyl group, preferably a C1_lo alkoxy-carbonyl group (e. g., methoxycarbonyl, ethoxycarbonyl, propoxycarbonyl, butoxycarbonyl), an aryloxycarbonyl group, preferably C6_1, aryloxycarbonyl group (e. g., phenoxycarbonyl), and an aralkyloxycarbonyl group , preferably a C,_19 aralkyloxy-carbonyl group ( a . g . , benzyloxycarbonyl).
The amidated carboxyl group represented by B is exemplified by an optionally substituted carbamoyl group represented by the formula: -CON( R1 ) ( RZ } wherein R1 and R2 independently are a hydrogen atom, an optionally substituted hydrocarbon group or an optionally substituted 5- to 7-membered heterocyclic group.
The hydrocarbon group of the optionally substituted hydrocarbon group represented by R1 or RZ is exemplified by the above-mentioned alkyl group, preferably an alkyl group having 1 to 10 carbon atoms (e. g., methyl, ethyl, propyl, isopropyl,butyl,isobutyl,sec-butyl,tert-butyl, pentyl, isopentyl, neopentyl, hexyl, heptyl, octyl, nonyl, decyl}, an alkenyl group, preferably an alkenyl group having 2 to 10 carbon atoms (e.g., allyl, vinyl, 2-penten-1-yl, 3-penten-1-yl, 2-hexen-1-yl, 3-hexen-1-yl, 2-cyclohexenyl, 2-cyclopentenyl, 2-methyl-2-propen-1-yl, 3-methyl-2-buten-1-yl), an aryl group, preferably aryl group having 6 to 14 carbon atoms ( a . g . , phenyl , naphthyl , anthryl) , and an aralkyl group, preferably aralkyl groups having 7 to 19 carbon atoms ( a . g . , benzyl , naphthylethyl , trityl). These hydrocarbon groups may be substituted by 1 to 3 substituents selected from ( i ) a halogen atom ( a . g . , fluorine , chlorine , bromine , iodine ) , ( ii ) a hydroxyl group , (iii} an alkoxy group having 1 to 6 carbon atoms (e. g., methoxy, ethoxy, propoxy, butoxy, text-butaxy, pentyloxy, hexyloxy), (iv) an amino group which may be substituted by an alkyl group having 1 to 6 carbon atoms such as methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, tert-butyl,pentyl,iso-pentyl,neo-pentyl and hexyl(e.g., amino, methylamino, ethylamino, dimethylamino, diethylamino, dipropylamino), (v) an amino group which may be substituted by an acyl group such as a C1_lo alkanoyl group (e.g., acetylamino, propionylamino, benzoylamino), (vi) a carbamoyl group which may be substituted by an alkyl group having 1 to 6 carbon atoms (e. g., carbamoyl, i2 methylcarbamoyl, dimethylcarbamoyl, diethylcarbamoyl), ( vii ) a C1_6 alkoxy-carbonyl group ( a . g . , methoxycarbonyl , ethoxycarbonyl, propoxycarbonyl), (viii) a mono- or di-alkoxyphosphoryl group (e. g. a mono- or di-C1_s alkoxyphosphoryl group such as dimethoxyphosphoryl, diethoxyphosphoryl, ethylenedioxyphosphoryl), (ix) a mono- or di-alkoxyphosphorylalkyl group (e.g. a mono- or di-Cl_6 alkoxyphosphoryl-C1_3 alkyl group such as methoxyphosphorylmethyl, ethoxyphosphorylmethyl, methoxyphosphorylethyl, ethoxyphosphorylethyl, dimethoxyphosphorylmethyl, diethoxyphosphorylmethyl, dimethoxyphosphoryethyl, diethoxyphosphorylethyl), (x) a moiety:

-CH2 - P ~ (CHZ) P
0 0~
wherein p is an integer from 2 to 4 , ( xi ) a phosphono group, (xii) the above-mentioned aromatic heterocyclic group, etc.
The 5- to 7-membered heterocyclic group of the optionally substituted 5- to 7-membered heterocyclic group represented by R1 or R2 is exemplified by a 5- to 7-membered heterocyclic group containing a sulfur, nitrogen or oxygen atom, 5- or 6-membered heterocyclic groups containing 2 to 4 nitrogen atoms, and 5- or 6-membered heterocyclic groups containing 1 or 2 nitrogen atom( s ) and a sulfur or oxygen atom. These heterocyclic groups may be condensed with a 6-membered ring containing 2 or fewer nitrogen atoms, a benzene ring or a 5-membered ring containing a sulfur atom.
As a substituent of the substituted 5- to 7-membered heterocyclic group represented by Rl and Rz, there may be used 1 to 4 of the same substituents as those for the substituted hydrocarbon group represented by R1 and RZ

above.
Preferable examples of the 5- to 7-membered heterocyclic group represented by R1 and RZ include 2-pyridyl, pyrimidyl, pyrazinyl, pyridazinyl, pyrazolyl, imidazolyl,thiazolyl,oxazolyl,tetrazolyl,thiadiazolyl, oxadiazolyl, triazinyl, triazolyl, thienyl, pyrrolyl, pyrrolinyl, furyl, pyrrolidinyl, benzothienyl, indolyl, imidazolidinyl, piperidyl, piperidino, piperazinyl, morpholinyl, morpholino, pyrido[2,3-d]pyrimidyl, benzopyranyl, 1,8-naphthyridyl, quinolyl, thieno[2,3-b]pyridyl.
The moiety: -N(R1) (R2) may form a 5- to 7-membered ring by binding together with Rl and R2. Such rings include morpholine, piperidine, thiomorpholine, homopiperidine, piperidine, pyrrolidine, thiazolidine and azepine.
The substituted alkyl group as preferable examples of the optionally substituted hydrocarbon group represented by R' and R2 include trifluoromethyl, trifluoroethyl, difluoromethyl, trichloromethyl, 2-hydroxyethyl, 2-methoxyethyl, 2-ethoxyethyl, 2,2-dimethoxyethyl, 2,2-diethoxyethyl, 2-pyridylmethyl, 3-pyridylmethyl, 4-pyridylmethyl, 2-(2-thienyl)ethyl, 3-(3-furyl)propyl, 2-morpholinoethyl, 3-pyrrolylbutyl, 2-piperidinoethyl, 2-(N,N-dimethylamino)ethyl, 2-(N-methyl-N-ethylamino)ethyl, 2-(N,N-diisopropylamino)ethyl, 5-(N,N-dimethylamino)pentyl, N,N-dimethylcarbamoylethyl,N,N-dimethylcarbamoylpentyl, ethoxycarbonylmethyl, isopropoxycarbonylethyl, tert-butoxy-carbonylpropyl, 2-diethoxyphosphorylethyl, 3-dipropoxyphosphorylpropyl, 4-dibutoxyphosphorylbutyl, ethylenedioxyphosphoryimethyl, 2-phosphonoethyl and 3-phosphonopropyl. The preferable substituted aralkyl groups include 4-chlorobenzyl, 3-(2-fluorophenyl)propyl, 3-methoxybenzyl, 3,4-dimethoxyphenethyl, 4-ethylbenzyl, 4-(3-trifluoromethylphenyl)butyl, 4-acetylaminobenzyl, 4-dimethylaminophenethyl, 4-diethoxy-phosphorylbenzyl and 2-(4-dipropoxyphosphorylmethylphenyl)ethyl. The preferable substituted aryl groups include4-chlorophenyl, 4-cyclohexylphenyl, 5,6,7,8-tetrahydro-2-naphthyl, 3-trifluoromethylphenyl, 4-hydroxyphenyl, 3,4,5-trimethoxyphenyl, 6-methoxy-2-naphthyl, 4-(4-chlorobenzyloxy)phenyl, 3,4-methylenedioxyphenyl, 4-(2,2,2-trifluoroethoxy)phenyl, 4-propionylphenyl, 4-cyclohexanecarbonylphenyl, 4-dimethyl-aminophenyl, 4-benzoylaminophenyl, 4-diethoxycarbamoylphenyl, 4-tert-i0 butoxycarbonylphenyl, 4-diethoxyphosphorylphenyl, 4-diethoxyphosphorylmethylphenyl, 4-(2-diethoxyphosphorylethyl)phenyl, 2-diethoxyphosphorylmethylphenyl, 3-diethoxyphosphorylmethylphenyl, 4-dipropoxyphosphorylphenyl, 4-(2-phosphonoethyl}phenyl, 4-phosphonomethylphenyl and 4-phosphonophenyl. The preferable substituted 5- to 7-membered heterocyclic groups include 5-chloro-2-pyridyl, 3-methoxy-2-pyridyl, 5-methyl-2-benzothiazolyl, 5-methyl-4-phenyl-2-thiazolyl, 3-phenyl-5-isoxazolyl, 4-(4-chlorophenyl}-5-methyl-2-oxazolyl, 3-phenyl-1,2,4-thiadiazol-5-yl, 5-methyl-1,3,4-thiadiazol-2-y1,5-acetylamino-2-pyrimidyl, 3-methyl-2-thienyl, 4,5-dimethyl-2-furanyl and 4-methyl-2-morpholinyl.
With respect to Compound ( I ) , ring A is preferably a benzene ring which may be substituted by 1 or more, more preferably 1 or 2 substituents selected from halogen atoms , an optionally substituted alkyl group, an optionally substituted hydroxyl group, an optionally substituted mercapto group and/or an optionally substituted amino group.
More preferably, ring A is a benzene ring which may be substituted by 1 or 2 substituents selected from the above-mentioned halogen atom, an alkyl group having 1 to 10 carbon atoms ( furthermore preferably 1 to 5 carbon atoms ) , an alkoxy group having 1 to 10 carbon atoms (furthermore preferably 1 to 5 carbon atoms), an alkylenedioxy group represented by the formula: -O-(CHZ)n-O- wherein n is an integer from 1 to 3, and/or an alkylthio group having 1 to 10 carbon atoms (furthermore preferably 1 to 5 carbon 5 atoms).
Most preferably, ring A is a benzene ring which may be substituted by an alkylenedioxy group represented by the formula : -O- ( CHZ ) n-O- wherein n is an integer from 1 to 3 .
R is preferably a hydrogen atom, an alkyl group 10 having 1 to 6 carbon atoms ( a . g . methyl , ethyl ) or a phenyl group.
B is preferably an alkoxycarbonyl group or a group represented by the formula : -CON ( Rl ) ( RZ ) wherein Rl and R2 independently are a hydrogen atom, an optionally 15 substituted hydrocarbon group or an optionally substituted 5- to 7-membered heterocyclic group.
With respect to R1 and RZ above , R1 is preferably a hydrogen atom or an alkyl group having 1 to 10 carbon atoms ( a . g . methyl , ethyl , propyl ) , and RZ is preferably a phenyl or phenyl-C1_3 alkyl group which may be substituted by a halogen atom (e. g. fluorine, chlorine, bromine), a C1_6 alkoxy (e.g. methoxy, ethoxy), a mono- or di-alkoxyphosphoryl (preferablly a mono- or di-C1_s alkoxyphosphoryl such as dimethoxyphosphoryl, diethoxyphosphoryl), a mono- or di-alkoxyphosphorylalkyl (preferablly a mono- or di-Cl_6 alkoxyphosphoryl-C1_3 alkyl such as dimethoxyphosphorylmethyl, diethoxyphosphorylmethyl) wherein dialkyl groups of these di-C1_6 alkoxy group may bind together to form a Cl_6 alkylene group or a C1_6 alkoxycarbonyl (e. g. methoxycarbonyl, ethoxycarbonyl) , or a 5- or 6-membered heterocyclic group ( a . g . pyridyl ) which may be substituted by a phenyl and that contains 1 or 2 nitrogen atom( s ) or a nitrogen atom and a sulfur atom.
More preferable example of R1 and Rz is a hydrogen atom, and a phenyl group substituted by a mono- or di-C1_6 alkoxyphosphoryl-C1_, alkyl, respectively (e.g. 4-diethoxyphosphorylmethylphenyl).
With respect to Compound (I), X is -CH(OH)- or -CO-, preferablly -CO-.
With respect to Compound (I), k is 0 or 1, and k' is 0, 1 or 2, preferablly k is 1, and k' is 0.
Compound (I) is preferably an optically active benzothiepine derivertives represented by the formula (II):
/Og4 _..
~CONH ~ ~ Cgzp ''' ,. 11\Ogs_ (II) C ,t s o 0 g3 wherein R' is a C1_6 alkyl group; and R' and R' are independently a C1_6 alkyl group or bind together to form a Cl_6 alkylene group .
The C1_6 alkyl group represented by R', R' or RS in the Compound (II) is exemplified by alkyl groups such as methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl,tert-butyl,pentyl,isopentyl,neo-pentyl and hexyl, and preferably a Cl_~ alkyl group. R4 and RS may bind together to form a C1_6 alkylene group. In this case, a moiety:
-p~0g 4 .',~~'~
n\
a may represent a moiety:

P\ /(CHZ)p wherein p is an integer from 2 to 4.

Preferable groups for R', R' and RS include alkyl groups having 1 to 4 carbon atoms such as methyl and ethyl.
The compound (II) is an optically active compound of the (2R,4S) configuration, and contains substantially no compound of the (2S,4R) configuration. The compound ( II ) of which optical purity is nearly 100% is preferable.
Most preferably, the compound ( II ) is, for example, (2R,4S)-(-)-N-[4-(diethoxyphosphorylmethyl)phenyl]-1,2,4,5-tetrahydro-4-methyl-7,8-methylenedioxy-5-oxo-3-benzothiepine-2-carboxamide {hereinafter also referred to as compound A ) or its salt . The compound A is represented as below.
CONH ~ ~ CHIP (0) (OC2H5) 2 S
p -CH3 The salt of Compound (I) is preferably a pharmaceutically acceptable salt. Pharmaceutically acceptable salts include salts with inorganic or organic bases , salts with inorganic or organic acids , and basic or acidic amino acids. Inorganic basic salts include alkali metal salts (e.g., sodium salts, potassium salts) and alkaline earth metal salts (e. g., calcium salts, magnesium salts). Such organic basic salts include the salts with trimethylamine, triethylamine, pyridine, picoline, N,N-dibenzyl-ethylenediamine or diethanolamine. Such inorganic acidic salts include the salts with hydrochloric acid, hydrobromic acid, hydroiodic acid, phosphoric acid, nitric acid and sulfuric acid. Such organic acidic salts include the salts with formic acid, acetic acid, trifiuoroacetic acid, oxalic acid, tartaric acid, fumaric acid, malefic acid, methanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid and citric acid. Such the salts with basic or acidic amino acids include the salts with arginine, lysine, aspartic acid and glutamic acid.
Compound (I) or its salt included in the pharmaceutical composition of the present invention can be produced by the method described in Japanese laid-open patent applications 232880/1991 (corresponding to EP-A-0376197),364179/1992(corresponding to EP-A-0460488), and 231569/1996(corresponding to EP-A-0719782) or a modification thereof.
It has been known that Compound (I) exhibits an excellent alkaline phosphatase inducing activity, and that it shows strong osteogenesis promoting effect (Japanese laid-open patent application 231569/1996, and so on).
However, it has not been known that Compound (I) normalizes the condition of the cartilage lesion by excellent chondrogenesis promoting effect, cartilage destruction suppressing effect and cartilage cell differentiation induction promoting effect. The present invention is made based on this fact. In this point the present invention is totally different from the above mentioned prior art. Therefore the pharmaceutical composition comprising Compound (I) of the present invention is useful for preventing or treating the cartilage disease, that it is especially effective for preventing or treating such disease before sickness reaches bone itself . Therefore the composition can be used not only for treating early stage of a cartilage defect, chronic rheumatoid arthritis and osteoarthritis, but also for preventing these desieses.
Since Compound (I) has potent cartilage destruction suppressing effect, chondorogenesis promoting effect, cartilage cell differentiation induction promoting effect, proteoglycan synthesis promoting effect, type II collagen synthesis promoting effect, metalloprotease I release suppressing effect and proteoglycan release suppressing WO 99!65474 PCT/JP99/03154 effect, and is more excellent in terms of clinically useful characteristics such as stability, absorption, bioavailability, it can be used for prevention and treatment of a cartilage destruction in a joint in any of various cartilage diseases such as cartilage defect, chronic rheumatoid arthritis involving a cartilage, osteoarthritis of a knee involving a cartilage as well as disorders related thereto, in mammals (e. g., human, rat, mouse, cat, dog, rabbit, cattle, pig, etc.).
The pharmaceutical composition comprising Compound ( I ) of the present invention can be administered orally or non-orally, as formulated with a pharmaceutically acceptable carrier, in the form of solid preparations such as tablets, capsules, granules and powders, or liquid preparations such as syrups and in~ectable preparations.
Pharmaceutically acceptable carriers are various organic or inorganic carrier substances in common use as pharmaceutical materials. They include excipients, lubricants, binders and disintegrants for solid preparations; and solvents, dissolution aids, suspending agents, isotonizing agents, buffers and soothing agents for liquid preparations. Other pharmaceutical additives such as preservatives, antioxidants, stabilizing agents, coloring agents and sweetening agents may be used as necessary.
Preferable excipients include lactose, sucrose, D-mannitol, starch, crystalline cellulose and light silicic anhydride.
Preferable lubricants include magnesium stearate, calcium stearate, talc and colloidal silica.
Preferable binders include binding cellulose, pregelatinized starch, sucrose, D-mannitol, dextrin, hydroxypropyl cellulose, hydroxypropylmethyl cellulose and polyvinylpyrrolidone.
Preferable disintegrants include starch, carboxymethyl cellulose, carboxymethyl cellulose calcium, croscarmellose sodium and carboxymethyl starch sodium, low-substituted hydroxypropyl cellulose.
Preferable solvents include water for injection, alcohol, propylene glycol, macrogol, sesame oil and corn 5 oil.
Preferable dissolution aids include polyethylene glycol, propylene glycol, D-mannitol, benzyl benzoate, ethanol,tris-aminomethane,cholesterol, triethanolamine, sodium carbonate and sodium citrate.
10 Preferable suspending agents include surfactants such as stearyltriethanolamine, sodium lauryl sulfate, lauryl-aminopropionic acid, lecithin, benzalkonium chloride, benzethonium chloride and monostearic glycerol;
and hydrophilic polymers such as polyvinyl alcohol, 15 polyvinylpyrrolidone, carboxymethyl cellulose sodium, methyl cellulose, hydroxymethyl cellulose, hydroxyethyl cellulose and hydroxypropyl cellulose.
Preferable isotonizing agents include sodium chloride, glycerol and D-mannitol.
20 Preferable buffers include buffer solutions of phosphates, acetates, carbonates and citrates.
Preferable soothing agents include benzyl alcohol.
Preferable preservatives include p-oxybenzoic acid esters, chlorobutanol, benzyl alcohol, phenethyl alcohol, dehydroacetic acid and sorbic acid.
Preferable antioxidants include sulfites and ascorbic acid.
The pharmaceutical composition of the present invention can be produced by dissolving or dispersing Compound ( I ) into an appropriate solvent and forming into microcapsules, spheres, rods, needles, pellets, films or the like, by an appropriate method.
In addition, the pharmaceutical composition of the present invention can also be a sustained-release preparation comprising a biodegradable polymer dispersed Compound ( I ) by a method such as that described in Japanese laid-open patent application 231569/1996.
The biodegradable polymer of the present invention is a polymer that is poorly soluble or insoluble in water and degradable in vivo in an appropriate period for treatment. Examples of such polymers include fatty acid polyesters such as polymers, copolymers and their mixture of one or more kinds of c~ -hydroxycarboxylic acids ( a . g . , lactic acid, gly-colic acid, 2-hydroxybutyric acid, 2-hydroxyvaleric acid, 2-hydroxy-3-methylbutyric acid, 2-hydroxycaproic acid, 2-hydroxyisocaproic acid, 2-hydroxycaprylic acid), hydroxydi-carboxylic acids (e. g., malic acid) and hydroxytricarboxyl-is acids (e. g., malic acid), lactic acid caprolactones, valerolactones, etc., and derivatives thereof ( a . g . , block polymers of polylactic acid, polyglycolic acid and polyethylene glycol), poly-cx-cyanoacrylates, polyalkylene oxalates (e. g., polytrimethylene oxalate, polytetramethy-lene oxalate), polyortho-esters, polyortho-carbonates, polycarbonates (e. g., polyethylene carbonate, polyethylene-propylene carbonate), polyamino acids (e.g., poly-r-benzyl-L-glutamic acid, poly-L-alanine, poly-?'-methyl-L-glutamic acid), hyarulonates, polystyrene, polymethacrylic acid, acrylic acid-methacrylic acid copolymers,polyamino acids, dakin stearate, ethyl cellulose, acetyl cellulose, vitro-cellulose, malefic anhydride copolymers, collagen, gelatin, fibrin and hydroxyapatite.
These biodegradable polymers may be in the form of homopolymers or copolymers of two or more kinds , or these mixtures.
Polymerization may be of the random, block or graft type.
Preferable biodegradable polymers in-clude aliphatic polyesters.
From the viewpoint of biodegradability and biocompati-bility, polymers and copolymers synthesized from one or more kinds of cx-hydroxycarboxylic acids are preferred. Specifically, copolymers synthesized from one or more kinds of lactic acid, glycolic acid, 2-hydroxybutyric acid, 2-hydroxyvaleric acid etc., or mixtures thereof are used.
The biodegradable copolymer for the present invention can be produced by commonly known methods such as that described in Japanese laid-open patent application 28521/1986 (EP172636), or a modification thereof.
Although the above-mentioned c~-hydroxycarboxylic acids may be of the D-, L- or D,L-configuration, the D,L-configura-tion is preferred.
Homopolymers of the above-mentioned a-hydroxycarboxylic acids include homopolymers of lactic acid, glycolic acid and 2-hydroxybutyric acid. The preferable a-hydroxycarboxylic acid is lactic acid.
Copolymers of the above-mentioned cx-hydroxycarboxylic acids include copolymers of glycolic acid and the other c~
-hydroxycarboxylic acids. Preferable cx-hydroxycarboxylic acids are lactic acid and 2-hydroxybutyric acid. Specifically, useful copolymers in-clude lactic acid-glycolic acid copolymers and 2-hydroxy-butyric acid-glycolic acid copolymers, with preference given to lactic acid-glycolic acid copolymers , etc.
The average molecular weight of these biodegradable polymers for the present invention is preferably chosen from the range of about 2 , 000 to 800 , 000 , more preferably about 5,000 to 200,000.
The weight-average molecular weight of a lactic acid homopolymer (hereinafter also referred to as polylactic acid) is preferably about 5 , 000 to 100 , 000 , more preferably about 6 , 000 to 50 , 000 . A polylactic acid can , for example, be synthesized by commonly known production methods such as that described in Japanese laid-open patent application 28521/1986 (EP172636).
The content ratio of lactic acid and glycolic acid in a lactic acid-glycolic acid copolymer is preferably about 100/0 to 50/50 (w/w) , and more preferably about 90/10 to 50/50 (w/w) . The weight-average molecular weight of the lactic acid-glycolic acid copolymer is preferably about 5,000 to 100,000, more preferably about 8,000 to 50,000.
The lactic acid-glycolic acid copolymer can be synthesized by a commonly known production method such as that described in Japanese laid-open patent application 28521/1986 (EP172636). The copolymer is preferably synthesized by catalyst-free dehydration polymerization condensation.
With respect to the 2-hydroxybutyric acid-glycolic ac-id copolymer, the content ratio is preferably such that glycolic acid accounts for about 40 to 70 mol%, and 2-hydroxybutyric acid accounts for the remaining portion.
The weight-average molecular weight of the 2-hydroxybutyric acid-glycolic acid copolymer is preferably about 5,000 to 100,000, more preferably about 8,000 to 50,000. The 2-hydroxybutyric acid-glycolic acid copolymer can be synthe-sized by a commonly known production method such as that described in Japanese laid-open patent application 28521/1986 (EP172636). The copolymer is preferably synthesized by catalyst-free dehydration polymerization condensation.
The above-described 2-hydroxybutyric acid-glycolic ac-id copolymer may be used in mixture with polylactic acid. When the 2-hydroxybutyric acid-glycolic acid copolymer is used in mixture with polylactic acid, the mixing ratio of 2-hydroxybutyric acid/glycolic acid is about 10/90 to 90/10 ( % by weight ) , preferably about 25/75 to 75/25 (% by weight).
In the present specification, weight-average molecular weight is defined as that based on polystyrene measured by gel permeation chromatography (GPC).
Measurements were taken using a GPC column KF804L X2 ( produced by Showa Denko ) and an RI monitor L-3300 ( produced by Hitachi Ltd.) with chloroform as a mobile phase.

The amount of biodegradable polymer is variable according to the strength of the pharmacological activity of Compound ( I ) , the speed and duration of drug release from the biodegradable polymer and so on, as long as the desired purpose is accomplished. For example, the biodegradable polymer is used in amounts about 0 . 2 to 10 , 000 times ( ratio by weight), preferably about 1 to 1,000 times, more preferably about 1 to 100 times, for the amount of the bioactive substance.
The pharmaceutical composition of the present invention can be produced by ordinary methods of producing a pharmaceutical composition, for example, it can be produced by dispersing a non-peptide osteogenetic promoting substance in a bio-degradable polymer, or by filling a non-peptide osteogenetic promoting substance in a previously shaped hollow biodegradable polymer.
Specifically, useful methods include the in-water drying method, the phase separation method, the spray drying method, and modifications thereof:
Example methods of producing microcapsules of the present invention are described below.
(1) In-water drying method (o/w method) In this method, an organic solvent solution comprising a biodegradable polymer is first prepared. The organic solvent used to produce the pharmaceutical composition of the present invention preferably has a boiling point of not higher than 120°C. Such organic solvents include halogenated hydrocarbons (e. g., dichloromethane, chloroform, chloroethane, dichloroethane, trichloroethane, carbon tetrachloride), aliphatic esters (e. g., ethyl asetate, butyl asetate), ethers (e. g., ethyl ether, isopropyl ether) and aromatic hydrocarbons (e. g., benzene, toluene, xylene). These solvents may be used in combination of two or more kinds in appropriate ratios. The organic solvent is pref-erably dichloromethane or acetonitrile. The organic solvent is more preferably dichloromethane. The concentration of biodegradable polymer in the organic solvent solution is normally chosen over the range of about 0 . O1 to 80% (w/w) , preferably about 0. 1 to 70% (w/w) , and more preferably about 5 1 to 60% (w/w), although varying depending on molecular weight of biodegradable polymer and organic solvent type, etc.
Compound ( I ) is added and dissolved into the organic solvent solution comprising the biodegradable polymer thus 10 obtained, if necessary after lyophilized or vacuum dried.
The amount of Compound (I) is about 0.001 to 90% (w/w), preferably about 0.01 to 80% (w/w), and more preferably about 0.1 to 50% (w/w), based on the concentration of biodegradable polymer in the organic solvent solution, 15 although varying depending on drug type, mechanism of action on cartilage destruction suppressing effect or chondorogenesis promoting effect, effect duration, etc..
The organic solvent solution thus prepared is then added to an aqueous phase to form an o/w emulsion using a 20 turbine type mechanical stirrer or the like. The volume of the aqueous phase is normally chosen from the range of about 1 to 10 , 000 times , preferably about 2 to 5 , 000 times , and more preferably about 5 to 2 , 000 times , for the volume of the oil phase.
25 An emulsifier may be added to the aqueous phase . The emulsifier may be any one as long as it is capable of forming a stable o/w emulsion. Examples of such emulsifiers include anionic surfactants, nonionic surfactants, polyoxyethylene castor oil derivatives, polyvinyl pyrrolidone, polyvinyl alcohol, carboxymethyl cellulose, lecithin, gelatin and hyaluronic acid. These may be used in combination as appropriate. The concentration of emulsifier in the aqueous phase is preferably about 0.001 to 20% (w/w) , more preferably about 0.01 to 10% (w/w) , and further more preferably about 0.05 to 5% (w/w).
Solvent evaporation from the oil phase can be achieved by commonly used methods , including the method in which the solvent is evaporated under normal or gradually reduced pressure during stirring using a propeller stirrer or magnetic stirrer, etc., and the method in which the solvent is evaporated while the degree of vacuum is adjusted using a rotary evaporator, etc.. The obtained microcapsules are separated by centrifugal method or filtration, after which they are washed with, for example, water or heptane, several times to remove free Compound ( I ) , emulsifier, etc. adhering to the microcapsule surface.
The microcapsules are then again dispersed in distilled water, etc. and lyophilized. To prevent particle flocculation during washing, antiflocculants: water-soluble sugars such as mannitol, lactol, glucose and starches ( e. g . , corn starch ) , amino acids such as glycine and alanine, and proteins such as gelatin, fibrin and collagen may be added.
In the above-described o/w method, microcapsules may be produced by the w/o/w method, in which Compound ( I ) is dispersed in an organic solvent solution comprising a biodegradable polymer.
(2) In-water drying method (w/o/w method) In this method, Compound ( I ) is first dissolved or dispersed in water to obtain a concentration specified above to yield an internal aqueous phase, if necessary with dissolving or suspending by adding a drug-retaining substance such as a protein ( a . g . , gelatin ) , seaweed ( a . g . , agar), polysaccharide (e. g., alginic acid), synthetic high-molecular substance (e. g., polyvinyl alcohol), basic amino acid (e.g., arginine, lysine) or the like. The internal aqueous phase may be supplemented with an organic acid such as acetic acid, oxalic acid or citric acid, an inorganic acid such as carbonic acid or phosphoric acid, an alkali metal hydroxide such as sodium hydroxide, a basic amino acid such as. arginine or lysine or a salt thereof ( a . g . , salts with organic acids such as acetic acid, oxalic acid, citric acid or salts with inorganic acids such as carbonic acid, phosphoric acid and hydrochloric acid) as a pH
regulator for keeping the stability and solubility of Compound (I) or its salt thereof. As a stabilizer for Compound ( I ) , there may be added a protein ( a . g . , albumin , gelatin), starch derivative (e. g. dextrin, pullulan), organic acid {e. g., citric acid), ethylenediaminetetraacetic acid alkali metal salt (e. g., sodium ethylenediamine-tetraacetate), sulfurous acid hydrogen alkali metal salt ( a . g. , sodium hydrogen sulfite ) , synthetic high-molecular substance (e. g., polyethylene glycol) or the like. Commonly preservatives may also be added p-oxybenzoates (e. g., methyl paraben, propyl paraben), benzyl alcohol, chlorobutanol and thimerosal.
The additional amount of compound ( I ) is about 0 . 001 to 90%
(w/w), preferably about 0.01 to 80% (w/w), and more preferably about 0.1 to 50% (w/w), although varying depending on drug type, mechanism of action on cartilage destruction suppressing effect or chondorogenesis promoting effect or effect duration, etc.
The obtained internal aqueous phase is added to a solution (oil phase) containing the biodegradable polymer, followed by emulsifying treatment, to yield a w/o emulsion.
This emulsification is achieved by a known dispersing methods which include the intermittent shaking method, the method using a mixer such as a propeller shaker or a turbine shaker, the colloidal mill method, the homogenizer method and the ultra-sonication method. The above-described solution (oil phase) containing the biodegradable polymer is a solution prepared by dissolving the biodegradable polymer in an organic solvent. This solvent may be any solvent as long as its boiling point is not higher than about 120°C and it is immiscible with water. Such solvents include halogenated hydrocarbons (e. g., dichloromethane, chloroform, chloroethane, dichloroethane, trichloroethane, carbon tetrachloride), aliphatic esters ( a . g . , ethyl acetate , butyl acetate ) , ethers ( a . g . , ethyl ether, isopropyl ether) and aromatic hydrocarbons (e. g., benzene , toluene , xylene ) . These solvents may be used in combination of two or more kinds in appropriate ratios . To prevent particle flocculation during washing, antiflocculants: water-soluble sugars such as mannitol, lactol, glucose and starches (e. g., corn starch), amino acids such as glycine and alanine, and proteins such as gelatin, fibrin and collagen may be added.
The produced w/o emulsion is then added to an aqueous phase to yield a w/o/w emulsion, from which the oil phase solvent is evaporated off, to yield microcapsules. The specific procedure for this production is the same as that described in (1) above.
(3) Phase separation method In this method, a coacervating agent is gradually added to the above-described w/o emulsion under the stirring to precipitate and solidify the biodegradable polymer. The coacervating agent can be used silicon oil, vegetable oils and fats (e.g. , sesame oil, soybean oil, corn oil, cotton seed oil, coconut oil, linseed oil), mineral oils , hydrocarbons ( a . g . , n-hexane , n-heptane ) as long as it is a polymeric, mineral oil or vegetable oil compound which can be mixed with the solvent of the biodegradable polymer and which does not dissolve the polymer for encapsulation. These may be used in combination of two or more kinds.
The obtained microcapsules are, after filtration and separation of them, repeatedly washed with heptane, etc .
to remove the coacervating agent . The free drug and solvent are then removed by using the same manner as in-water drying method. To prevent particle flocculation during washing, antiflocculants: water-soluble sugars such as mannitol, lactol, glucose and starches (e. g., corn starch), amino acids such as glycine and alanine, and proteins such as gelatin, fibrin and collagen may be added.

(4) Spray drying method For producing microcapsules by this method, the above-described w/o emulsion is sprayed via a nozzle into the drying chamber of a spray drier to volatilize the organic solvent and water in the fine droplets in a very short time, and microcapsules are obtained. The nozzle is exemplified by the double-fluid nozzle, pressure nozzle and rotary disc nozzle. To prevent microcapsule flocculation, an aqueous solution of the above-described antiflocculant may be sprayed via another nozzle, while the w/o emulsion is sprayed. The microcapsules thus obtained may be warmed under reduced pressure to facilitate the removal of the water and solvent contained them.
When microcapsules are used as an injectable suspension, for instance, their particle size is chosen over the range from about 0.1 to 300 ,um of average particle diameter, as long as the requirements concerning the degree of dispersion and needle passage are met . Preferably, the particle size is about 1 to 150 /.cm, more preferably about 2 to 100 ;um.
Methods of preparing microcapsules as a sterile preparation include, but are not limited to, the method in which the entire production process is sterile, the method in which gamma rays are used as sterilant , and the method in which an antiseptic is added.
In addition to the above-described microcapsules, the sustained-release preparation of the present invention can be produced by dissolving a biodegradable polymer dispersed Compound (I) and forming the solution into spheres , rods , needles , pellets , films or the like , by an appropriate method.
In addition, the sustained-release preparation of the present invention can also be produced by pulverizing to appropriate particle size a biodegradable polymer dispersed Compound ( I ) by a method such as that described in Japanese laid-open patent applications 234656/1994, which employs a turbo counter jet mill pulverizes or an ultrasonic jet pulverizes. Specifically, Compound (I) is added to an organic solvent containing the biodegradable polymer, and dissolved therein. The solid solution 5 obtained by vacuum drying is then coarsely pulverized $nd sieved, followed by solvent removal, after which the coarse particles are pulverized to controlled particle size using an ultrasonicjet pulverizes to yieldthesustained-release preparation of the present invention.
10 On the above mentioned preparation of the pharmaceutical composition, the content ratio of Compound ( I ) based on the pharmaceutical composition is about 0 . O1 to 95% (w/w), and preferbly about 0.1 to 20% (w/w).
However, the pharmaceutical composition of the 15 present invention can be administered as an oral agent , a non-oral agent for local administration (e. g., injectable preparations of intramuscular, subcutaneous, organs or joints, etc., solid preparations such as, indwellable preparations, granules and powders, liquid preparations 20 such as suspensions, and ointments) is more preferable.
The practical injectable preparation can be prepared as aqueous suspension by suspending Compound ( I ) in water, along with a dispersing agent (e.g. , surfactants such as Tween 80 and HCO-60, polysaccharides such as 25 carboxymethyl cellulose, sodium alginate and hyarulonic acid, and polysorbate), a preservative (e. g., methyl paraben, propyl paraben), an isotonizing agent (e. g., sodium chloride,mannitol,sorbitol,glucose),buffer(e.g.
calcium carbonate), pH adjusting agent (e. g. sodium 30 phosphate, potassium phosphate), etc., and may be also prepared as an oily suspension by dispersing Compound ( I ) in a vegetable oil such as sesame oil or corn oil with or without a phospholipid such as lecithin, or a moderatelength fatty acid triglyceride (e. g., MIGLYOL
812 ) .
When the the pharmaceutical composition of the present invention is administered locally to a joint of a patient of osteoarthritis, the pharmaceutical composition may be the preperation in which Compound ( I ) is dispersed in the injectable hyalronic acid pharmaceutical composition (e.g., KAKENSEIYAKU, trade name: *ALTZ). The hyaluronic acids can be used as its pharmaceutically acceptable salts. The salts include alkali metal salts (e.g. , sodium salts , potassium salts ) and alkaline earth metal salts (e.g., calcium salts, magnesium salts ), prepherably sodium salts. The weight-average molecular weight of the hyaluronic acid or a salt thereof about 200 , 000 to 5 , 000 , 000 , preferably about 500 , 000 to 3 , 000 , 000 , more preferably about 700,000 to 2,500,000.
The concentration of Hyaluronic acid or the sodium salts in the dispersion medium dispersing Compound ( I ) is less than 1% (W/v) , preferably about 0.02 to I% (W/v) , more preferably about 0.1 to 1%(W/v) , because its viscosity is proper to administrate by injection.
The dispersion medium can include pH regulators, local anesthetics, antiiotics, dissolution aids, isotonizing agents, adsorption preventing agents, glycosaminoglycans, polysaccharides and the like, which are conventionaly used in this field.
Preferable examples include mannitol, sorbitol, sodium chloride, glycine, ammonium acetate or water-soluble protein which is substantally inactive in the body.
Preferable glycosaminoglycan include hyaluronic acid. condroitin, condroitin sulfate A, condroitin sulfate C, dermatan sulfate, heparin, heparan sulfate, and the like .
Preferable water-soluble protein include, which is disolved in water or physiological salt solution, include humanserum albmin,humanserum globulin, collagen, gelatin, etc.
Preferable pH regulators include glycine, ammonium acetate, citric acid, hydrochloric acid, sodium hydroxyde, etc.
Preferable local anesthetics include chlorobutanol, lidocine hydrochloride, etc.
Preferable antibiotics include gentamicin, etc.
Preferable dissolution aids include glycerin, polyethyleneglycol-400, etc.
Preferable isotonizing agents include mannitol, sorbitol, sodium chloride, etc.
Preferable adsorption preventing agents include polyoxyethylene sorbitan monooleate, etc.
The dose of the water-soluble protein may be about 0. 05 to 50 mg, preferably about 0. 5 to 20 mg, more preferably about 0.75 to 10 mg per a injectable preperation.
The phosphate or its salts (e.g. , sodium phospate, pottasium phosphate ) can enhance the activity of the pharmaceutical composition of the present invention. The concentration of sodium phosphate or potassium phosphate in the in~ectable preparation is about 0.1 mM to 500 mM, preferably about 1 mM to 100 mM.
The preferable preparation of the present invention is as follows.
(A) a lactic acid-glycolic acid copolymer:
wherein the ratio of lactic acid/glycolic acid is about 90/10 to 50/50 (w/w) and the weight-average molecular weight is about 8000 to 50000, (B) Compound (I}: (2R,4S)-(-)-N-[4-(diethoxyphosphorylmethyl)phenyl]-1,2,4,5-tetrahydro-4-methyl-7,8-methylenedioxy-5-oxo-3-benzothiepine-2-carboxamide, and (C) phosphoric acid or its salt: sodium phosphate.
The content ratio of ( B ) based on ( A ) is about 5 to 30% ( w/w ) . The content ratio of ( C ) based on ( A ) and (B) is about 0.1 to 20% (w/w).
The pharmaceutical composition of the present invention is preferably a suspension as described above.

The pharmaceutical composition of the present invention is preferably in the form of fine particles . This is because said pharmaceutical composition is less likely to cause excess pain to the patient when administered through an injection needle for ordinary subcutaneous or intramuscular injection.
The pharmaceutical composition of the present invention is preferably an injectable preparation.
Methodsof preparing the pharmaceutical composition of the present invention as a sterile preparation include, but are not limited to, the method in which the entire production process is sterile, the method in which gamma rays are used as sterilant, and the method in which an antiseptic is added.
Since the pharmaceutical composition of the present invention has excellent chondrogenesis promoting effect, cartilage destruction suppressing effect and cartilage cell differentiation induction promoting effect, it can be used in prevention and treatment of a cartilage disease ( for example, chronic rheumatoid arthritis, osteoarthritis of knee. Among these diseases, a lesion fixed and covered frequently with a brace is especially applicable to a sustained-release preperation formulation according to the present invention, since it requires a promoted cure continuously only by a single administration rather than frequent administrations.
The pharmaceutical composition according to the present invention may be used also in combination with other pharmaceuticals for treating articular diseases. For example, when Compound (I) is used as a cartilage destruction preventing/cartilage cell differentiation induction promoting agent, other pharmaceuticals for treating articular diseases may be used in combination.
Examples of such pharmaceuticals employed in combination are antiinflammatory steroidal agents (e. g. , prednisolone, hydrocortisone, methylprednisolone, dexamethasone, betamethasone etc.), non-steroidal antiphlogistic/analgesic agents (e. g., indomethacin, diclofenac, loxoprofen, ibuprofen, aspirin, piroxicam, sulindac, etc.) or hyaluronic acid formulations (e. g., sodium hyaluronate, etc.).
The pharmaceutical composition according to the present invention is used as a safe and highly potent formulation suitable to be used in prevention and treatment of a cartilage disease as well as repair and regeneration of a cartilage tissue. For example, it can exert a chondrogenesis promoting effect locally and efficiently, and improves the quality of life in a patient whose routine activities are affected adversely by a pain due to the wornout or the destruction of an articular cartilage.
The dose of the pharmaceutical composition of the present invention may be an pharmacologically effective amount of Compound (I), although depending on type of Compound (I), the patient's condition, the way of administration, releasing time of the active ingredient, and subject hosts, etc. For example, when the pharmaceutical composition of the present invention is administered orally, administration dose is in the range of about 5 to about 1000 mg, preferably about 30 to about 600 mg, based on the active ingredient content (e. g., compound (I)), per adult (weighing 50 kg), and it can be administered once to three times a day dividedly. When the pharmaceutical composition of the present invention is administered non-orally, it may be administered at about 0.1 to 500 mg, preferably about 1 to 50 mg, based on the active ingredient content ( a . g . , compound ( I ) ) , per adult (weighing 50 kg) , and it can be administered once to three times a day dividedly. When the sustained-release preperation of the present invention is administered non-orally, Compound ( I ) may be released about 0 .1 to about 100 mg a week.
In addition, Compound ( I ) can safely be used because of its low toxicity. For example, 500 mg/kg/day of Compound A given orally for 2 weeks in rats caused no abnormal findings. Since Compound A is more excellent in terms of absorption especially after oral administration when 5 compared with a corresponding racemate, it can advantageously be used in an oral formulation.
Brief Description of the Drawings Figure 1 indicates the results of the examination 10 for the effect on Interleukin-1-stimulated type-I
collagenase release in rabbit cartilage cell observed in Experiment 1.
Figure 2 indicates the results of the examination for the effect on Interleukin-1-stimulated proteoglycan 15 release in rabbit cartilage cell observed in Experiment 2.
Figure 3 indicates the results of the examination for the effect on Interleukin-1-stimulated proteoglycan synthesis observed in Experiment 3.
20 Best Mode for Carrying out the Invention The present invention is hereinafter described in more detail by means of the following working examples, which are not to be construed as limitative.
Compound: (ZR,4S)-(-)-N-[4-25 (diethoxyphosphorylmethyl)phenyl]-1,2,4,5-tetrahydro-4-methyl-7,8-methylenedioxy-5-oxo-3-benzothiepine-2-carboxamide (hereinafter also referred to as Compound A) is prepared according to the method described in Example 1 of Japanese Patent Laid Open Publication 231569/1996.
Examples Example 1 About 8 g of a lactic acid-valerolactone copolymer (PLV 2500ML, produced by Taki Chemical, hereinafter also referred to as PLV) or a glycolic acid-caprolactone co-polymer (PGC 2500MG, produced by Taki Chemical, hereinafter also referred to as PGC ) was placed in a centrifugal tube and heated to about 50 °C in a water bath. About 80 mg of compound A was mixed in each tube, followed by uniform dispersion, to yield an ointment preparation, which was stored at a cold place.
Example 2 500 mg of microcapsule prepared in the same manner as Example 1 of Japanese Patent Laid Open Publication 263545/1997 was uniformly dispersed in two test tubes of fibrinogen solution for Tisseel (produced by Nippon Zoki Pharmaceutical). Thrombin solution of two test tubes for Tisseel were gradually added. Subsequently, the mixture was immediately aspirated into a plastic syringe. The syringe was kept standing at 37 °C for 30 minutes to solidify the content. After solidification, the content was extruded from the syringe tip and cut using a razor into pellets about 200 lcl in volume.
Example 3 4 mg of Compound A was filled in the bone defect filler hollow hydroxyapatite (Boneceram P, produced by Sumitomo Pharmaceuticals, 3 mm diameter, 14 mm length, 1 mm pore size). Both ends of the hollow were sealed with clay.
Example 4 To microcapsule prepared in the same manner as Example 1 of Japanese Patent Laid Open Publication 263545/1997 which contains compound A (content ratio 4%), 20~ pulverized gelatin (produced by Nitta Gelatin) was added, to yield a microcapsule-containing tablet preparation 5.5 mm in diameter and 125 mg in weight.
Example 5 Microcapsule containing compound A (content ratio 10~ ) was prepared in the same manner as Example 1 of Japanese Patent Laid Open Publication 263545/1997, except that PLGA
having a lactic acid-glycolic acid content ratio of 85/15 (mold) and weight-average molecular weight of 14,900 (produced by Wako Pure Chemical Industry). Mean particle size was 31 lam.
Example 6 A dichloromethane solution containing 2.4 g of PLGA
( produced by Wako Pure Chemical Industry ) whose the lactic acid/glycolic acid content ratio is 85/15 and the weight-average molecular weight is 14, 900 and 0.1 g of the compound A was prepared in the same manner as Example 1 of Japanese Patent Laid Open Publication 263545/1997. And 0.2 g of estradiol was added into the solution. Further PVA solution was put into the solution to obtain O/W
emulsion. Microcapsule containing the compound A and estradiol was prepared. The mean particle size was 27 a m.
Example 7 Preparation of capsule:
2 mg of hyaluronic acid (Q. P. Corporation, trade name: HATMKW7001, MW: about 2,100, 000), 2 mg of sodium chloride and 1 mg of Tween 80 ( NIKKO Chemical , trade name *RHEODOL*TW-0120 INJ NF) were weighed and dissolved in 1 ml of distilled water for injection to form a dispersion medium.
Separately, a microcapsule containing Compound A at 10 ~ was prepared by the method described in Example 6 in Japanese Patent Application Laid-Open 263545/1997. 10 mg of this microcapsule was weighed and dispersed in 0.2 ml of the dispersion medium described above to form an aqueous suspension. The suspension was used as a pharmaceutical composittion for prevention and treatment of cartilage disease which contained Compound A as an active ingredient .
Experiment 1 Effect on the release of type-I collagenase in rabbit cartilage cell stimulated by Interleukin-1 (IL-1):
A cartilage cell was prepared and cultured according to the method by Suzuki et al ( SHINSEIKAGAKUJIKKENKOZA 18 , Cell culture technology: 871-875, 1990). Thus, a Japanese WO 99/65474 PCT/.TP99/03154 white rabbit (male, 400 to 500 g) just after weaning was asphyxiated with C02 gas, and the whole body was sterilized with 70 ~ ethanol and then a thorax was isolated aseptically in a ventilated clean chamber. Each single costal cartilage/bone margin was isolated to obtain a cartilage piece free from circumferential soft tissues. This cartilage piece was cut into pellets using a scalpel, washed with Tyrode's solution free from Ca2' or Mg2', and then suspended in a Dulbeco's modification of Eagle's medium , (DMEM) containing 10 ~ calf fetal serum, and then inoculated into a 12-well microplate at the cell density of 4 x 10' cells/well. The cell was cultured for one week and the culture medium was exchanged with a serum-free medium, and then IL-1~ (30 ng/ml) and Compound A were added at predetermined concentrations . After 48 hours , the culture supernatant was recovered and the collagenase activity in the ' culture supernatant was determined using a commercial kit Type-I collagenase activity assay kit (COSMOBIO). The results are shown in Figure 1 ( * : p<0 . 05, Dunnet test ) . In Figure 1, the control is a sample containing no Compound A.
As evident from the results shown in Figure 1, Compound A exhibited type-I collagenase (metalloprotease I ) release suppressing effect, and had an inhibitory effect on a cartilage destruction due to arthritis.
Experiment 2 Effect on the release of proteoglycan in rabbit cartilage cell stimulated by Interleukin-1 A cartilage cell was prepared and cultured according to the method by Suzuki et al (SHINSEIKAGAKUJIKKENKOZA 18, Cell culture technology: 871-875, 1990). Thus, a Japanese white rabbit (male, 400 to 500 g) just after weaning was asphyxiated with COZ gas , and the whole body was sterilized with 70 ~ ethanol and then a thorax was isolated aseptically in a ventilated clean chamber. Each single costal cartilage/bone margin was isolated and circumferential soft tissues were removed as far as possible and then the margin between a bone and a cartilage was cut to obtain a cartilage piece free from other tissues. This cartilage piece was cut into pellets using a scalpel, and placed in Tyrode's solution free from Ca2' or Mgz+, and treated successively with EDTA (ethylenediaminetetraacetic acid) ( 0 . 1% , 20 minutes , twice ) , trypsin ( 0 .15% , 60 minutes ) and collagenase ( 0 . 2% , 3 hours ) at 37°C , whereby dispersing the cartilage cell. After washing the cell with the same Tyrode's solution, the cell was suspended in Dulbeco's modification of Eagle's medium (DMEM) containing 10 % calf fetal serum, and then inoculated into a 96-well microplate at the cell density of 1 x 10' cell/well. The cell was cultured for 4 days and then cultured in a serum-free/ ['SS ] sulfuric acid at 0 . 5 % ~Ci ( 1 x 106 dpm/well ) for 3 days, and then after washing twice it was cultured in [ssS]sulfuric acid-free medium. After 6 hours, IL-1~ (3 ng/ml) and Compound A were added again at predetermined concentration and then the culture was continued. 24 Hours after this addition, the culture supernatant was recovered and counted for the radioactivity. The cell was extracted with 4M guanidine hydrochloride and then subjected to the determination as described above. The results are shown in Figure 2 . In Figure 2 , the control is a sample containing no Compound A. "**" designates p<0.01 when compared with IL-1 free sample ( Student t-test ) . "##" designates p<0 . O1 when compared with IL-1-supplemented sample (Dunnet test).
As evident from the results shown in Figure 2, Compound A had a proteoglycan release suppressing effect, thus a cartilage destruction suppressing effect.
Experiment 3 Effect on proteoglycan synthesis in rabbit cartilage cell stimulated by Interleukin-1 (IL-1):
The cartilage cell prepared by the method similar to that in Experiment 1 was suspended in Dulbeco's modification of Eagle's medium (DMEM) and inoculated into a 96-well microplate at the concentration of 1 x 10' cells/well. After culturing for 4 days, the cell was treated with IL-1 (0.3 ng/ml} in the absence of serum for 24 hours . Subsequently, the cell was washed and cultured 5 for 48 hours in the presence of ['SS] sulfuric acid. After completion of the culture, the cell was washed with physiological saline, extracted with 4 M guanidine hydrochloride, and the resultant solution was counted for the radioactivity, whereby assaying the synthesis of 10 proteoglycan. The results are shown in Figure 3. In Figure 3, the control is a sample containing IL-1 (0.3 ng/ml) in the absence of test substance. Compound A was added at 10 NM simultaneously with ['SS ] sulfuric acid ( ( 1 ) in Figure 3 ) . Insulin-like growth factor ( IGF ) -I was added 15 at 10 ng/ml simultaneously with ['SS ] sulfuric acid ( ( 2 ) in Figure 3). (3) in Figure 3 indicates the addition of 10 ~.M of Compound A and 10 ng/ml of IGF-I which was simultaneous with the addition of ['SS ] sulfuric acid . In Figure 3 , the legends mean as follows: *P<0.05, **P<0.01 based on Dunnet 20 test when compared with control; ##p<0.01 based on Student t-test when compared with sample containing only IGF-I;
$$p<0.01 based on Student t-test when compared with sample containing only Compound A.
As evident from the results shown in Figure 3, 25 Compound A had proteoglycan synthesis promoting effect.
Experiment 4 A mouse cell line ATDCS (purchased from RIKAGAKU
KENKYUSHO ) was inoculated into a 6-well microplate at the density of 1 x 105 cells/well and cultured in DMEM/F-12 in 30 the presence of 5 ~ calf fetal serum and 10 ~,g/ml insulin.
On the 7th day of the culture, 10 N.M of Compound A was added and the culture was continued. The culture medium was replaced every 1 to 2 days. On the 14th day and the 21st day, the cell was recovered and subjected to the extraction 35 of mRNA using STAT60 (COSMOBIO) followed by RT-PCR method to quantify type II collagen mRNA expressed. As a control, actin mRNA expressed was determined.
The results are shown in Table 1. The control was the culture in the presence of 10 ~g/ml insulin.
Table 1 type II collagenmRNA/ actin mRNA

I4cn 2lat Control 0.0669 0.0325 IOI~M of Compound 0.1586 0.2876 A

As evident from the results shown in Table I, Compound A had type II collagen mRNA synthesis promoting effect.
Industrial Applicability The pharmaceutical composition of the present invention has potent cartilage destruction suppressing effect, chondorogenesis promoting and cartilage cell differentiation induction promoting effect, proteoglycan synthesis promoting effect, type II collagen synthesis promoting effect, metalloprotease I release suppressing effect and proteoglycan release suppressing effect, and is useful in prevention and treatment of cartilage disease.

Claims (15)

42

1. A pharmaceutical composition for prevention and/or treatment of a cartilage disease, which comprises a compound (I) of the formula:
wherein ring A is an optionally substituted benzene ring;
R is a hydrogen atom or an optionally substituted hydrocarbon group; B is an optionally esterified or amidated carboxyl group; X is -CH(OH)- or -CO-; k is 0 or 1; and k' is 0, 1 or 2, or its salt.

2. A pharmaceutical composition according to claim 1, wherein the ring A is a benzene ring which may be substituted by 1 or 2 substituents selected from the group consisting of a halogen, a C1-10 alkyl, a C1-10 alkoxy, an alkylenedioxy group of the formula: -O-(CH2)n-O- wherein n is an integer from 1 to 3 and a C1-10 alkylthio group: R
is a hydrogen atom, a C1-6 alkyl or a phenyl group; B is -CON(R1) (R2) wherein R1 is a hydrogen atom or a C1-10 alkyl group and R2 is a phenyl or a phenyl-C1-3 alkyl group which those groups may be substituted by a halogen, a C1-6 alkoxy, a mono- or di-C1-6 alkoxyphosphoryl, a mono- or di-C1-6 alkoxyphosphoryl-C1-3 alkyl wherein two alkyl groups of these di-C1-6 alkoxy group optionally may bind together to form a C1-6 alkylene group and C1-6 alkoxycarbonyl group.

3. A pharmaceutical composition according to claim 1, wherein the compound is an optically active Compound (II) of the formula;

wherein R3 is a C1-6 alkyl group; and R4 and R5 are independently a C1-6 alkyl group or bind together to form a C1-6 alkylene group.

4. A pharmaceutical composition according to claim 3, wherein R3, R4 and R5 are independently a C1-4 alkyl group.

5. A pharmaceutical composition according to claim 3, wherein the optically active compound (II) is (2R,4S)-(-)-N-[4-(diethoxyphosphorylmethyl)phenyl]-1,2,4,5-tetrahydro-4-methyl-7,8-methylenedioxy-5-oxo-3-benzothiepine-2-carboxamide.

6. A pharmaceutical composition according to claim 1, which is in the form of a sustained-release preparation comprising a biodegradable polymer.

7. A pharmaceutical composition according to claim 1, which is for local administration.

8. A pharmaceutical composition according to claim 1, which is for oral administration.

9. A pharmaceutical composition according to claim 1, which is for injection.

10. A pharmaceutical composition according to claim 1, which is for potent cartilage destruction suppressing agent, chondorogenesis promoting agent or cartilage cell differentiation induction promoting agent.

11. A pharmaceutical composition according to claim 1, which is for proteoglycan synthesis promoting agent, type II collagen synthesis promoting agent, metalloprotease I release suppressing agent or proteoglycan release suppressing agent.

12. A pharmaceutical composition according to claim 1, which is for proteoglycan synthesis promoting agent or metalloprotease I release suppressing agent.

13. A pharmaceutical composition according to claim 1, wherein the cartilage disease is a cartilage defect, chronic rheumatoid arthritis and osteoarthritis.

14. A method for prevention and/or treatment of a cartilage disease, which comprises administrating Compound (I) or its salts defined in claim 1 to mammal in need.

15. Use of Compound (I) or its salts defined in claim 1 for manufacturing a medicament for prevention and/or treatment of a cartilage disease.