KR19990022514A - Bone regeneration promoting pharmaceutical composition - Google Patents

Abstract

The present invention provides a pharmaceutical composition containing a non-peptide bone regeneration-promoting substance and a biodegradable polymer, which composition can be safely used as a prophylactic / treatment agent for various bone diseases (especially fractures).

Description

Bone regeneration promoting pharmaceutical composition

Bone disease (eg, fractures) can occur in all kinds of people due to various causes of sports and traffic accidents. And, because it takes a long time to treat these bone diseases, it significantly interferes with the normal life of the patient. Recently, the number of osteoporosis patients is increasing with the aging of the population. Thus, accidents of limb fractures associated with osteoporosis have increased significantly in proportion. Thigh neck fractures, in particular, require long-term inpatient treatment and often lead to complications in the body, including dementia caused by long-term inpatient treatment, which is a major social and economic problem. Discharge of fracture patients early from the hospital is an urgent task.

Fracture therapy is an injury treatment characterized by local incidence and progress. In general, various local factors act on the fracture site to enhance treatment in vivo. Such factors include peptide-type bioactive substances such as bone morphogenetic protein (BMP) and mutant growth factor (TGF). This substance is reported to promote bone regeneration in animal models of the National Academy of Sciences Bulletin, USA, Vol. 87, pp. 2200-2224 (1990), and Endocrinology, Vol. 124, 2991-2993 (1989).

As non-peptide-based bone regeneration-promoting substances, for example, prostaglandin A ₁ derivatives, vitamin D ₃ derivatives, benzylphosphinic acid derivatives, and phenolsulfophthalenoic acid derivatives have been reported.

Bioactive materials of the above mentioned peptide types are peptides or proteins whose molecular weight exceeds 5000 and lack stability because they are rapidly metabolized in vivo. In view of this, attempts have been made to prepare a formulation with satisfactory stability, but all have failed to achieve sufficient bone regeneration promoting activity, and no satisfactory formulation of such quality has been obtained (clinical orthopedic and related fields, Vol. 278). , Pages 274-285). In addition, the aforementioned non-peptide bone regeneration enhancing agents have not been clinically effective in terms of bone regeneration enhancing activity for treating fractures.

For this reason, there is a strong demand for high quality drugs for the treatment of fractures that are very stable, safe, active and clinically effective in the long-term treatment of fractures.

The present invention relates to a pharmaceutical composition having improved bone regeneration enhancing activity containing a non-peptide bone regeneration enhancing substance and a biodegradable polymer which is a medicament useful for the treatment and / or prevention of bone diseases.

The present inventors have conducted extensive research to solve this problem, and as a result, the bone of the non-peptide bone regeneration-promoting substance than the non-peptide bone regeneration-promoting substance and the biodegradable bone disease therapeutic agent containing biodegradable polymers is unexpectedly administered alone. It has been found that enhanced regenerative activity can satisfactorily improve fracture treatment. Based on these findings, the inventors continued to study to complete the present invention.

Therefore, the present invention

(1) a pharmaceutical composition containing a non-peptide bone regeneration-promoting substance and a biodegradable polymer,

(2) the pharmaceutical composition of (1) further containing phosphoric acid or a salt thereof,

(3) the pharmaceutical composition of the above (1), wherein the non-peptide bone regeneration-promoting substance is a nonsteroidal system,

(4) the pharmaceutical composition of (1) used for topical administration,

(5) the pharmaceutical composition of (1) used for the promotion of fracture treatment,

(6) the pharmaceutical composition of (1), which is a sustained release preparation,

(7) The pharmaceutical composition of (1), wherein the non-peptide bone regeneration-promoting substance is a compound represented by the following formula (I) or a salt thereof:

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, and X is -CH (OH)-or -CO-, k is 0 or 1, k 'is 0, 1 or 2),

(8) Ring A is halogen, C _1-10 alkyl, C _1-10 alkoxy, —O— (CH ₂ ) _n —O—, where n is 1 to 3 and C _1-10 alkylthio The pharmaceutical composition of the above (7), which is a benzene ring which may be substituted with one or two substituents selected.

(9) B is -CON (R ₁ ) (R ₂ ), wherein R ₁ and R ₂ are independently a hydrogen atom, an optionally substituted hydrocarbon group or an optionally substituted 5-7 membered heterocyclic group Pharmaceutical composition of the above (7),

(10) R ₁ is a hydrogen atom or a C _1-10 alkyl group, R ₂ is (i) halogen, C _1-6 alkoxy, mono- or di-C _1-6 alkoxyphosphoryl, mono- or di-C _{1 -6} alkoxyphosphoryl-C _1-3 alkyl,

Wherein p is 2 to 4 or a phenyl or phenyl-C _1-3 alkyl group which may be substituted by C _1-6 alkoxycarbonyl, or (ii) one or two nitrogen atoms which may be substituted by phenyl or The pharmaceutical composition of the above (9), which is a 5- or 6-membered heterocyclic group containing one nitrogen atom and one sulfur atom,

(11) the pharmaceutical composition of the above (7), wherein R is a hydrogen atom, a C _1-6 alkyl group, or a phenyl group;

(12) the pharmaceutical composition of (7), wherein k is 1 and k 'is 0;

(13) The pharmaceutical composition of the above (1), wherein the non-peptide bone regeneration enhancing substance is an optically active compound represented by the following general formula (II).

(Wherein R ₃ is a lower alkyl group and R ₄ and R ₅ are independently lower alkyl groups or combine with each other to form a lower alkylene group)

(14) The pharmaceutical composition of the above (13), wherein R ₃ , R ₄ and R ₅ are independently a C _1-6 alkyl group.

(15) The compound is (2R, 4S)-(-)-N- [4- (diethoxyphosphorylmethyl) phenyl] -1,2,4,5-tetrahydro-4-methyl-7,8- The pharmaceutical composition of the above (1), which is methylenedioxy-5-oxo-3-benzothiefine-2-carboxamide,

(16) the pharmaceutical composition of (1), wherein the weight ratio of the biodegradable polymer to the non-peptide bone regeneration enhancing substance is about 1 to 100 times;

(17) (2R, 4S)-(-)-N- [4- (diethoxyphosphorylmethyl) phenyl] -1,2,4,5-tetrahydro-4-methyl-7,8-methylenedioxy The pharmaceutical composition according to the above (1), which contains -5-oxo-3-benzothiine-2-carboxamide and a biodegradable polymer.

(18) the pharmaceutical composition of (17) further containing phosphoric acid or a salt thereof;

(19) The pharmaceutical composition of (18), wherein the phosphoric acid or a salt thereof is sodium phosphate,

(20) (2R, 4S)-(-)-N- [4- (diethoxyphosphorylmethyl) phenyl] -1,2,4,5-tetrahydro-4-methyl-7 for biodegradable polymers, The content ratio of 8-methylenedioxy-5-oxo-3-benzothiene-2-carboxamide is about 5 to 30% (w / w), and (2R, 4S)-(-)-N- [4 -(Diethoxyphosphorylmethyl) phenyl] -1,2,4,5-tetrahydro-4-methyl-7,8-methylenedioxy-5-oxo-3-benzothiene-2-carboxamide and The pharmaceutical composition of the above (17), wherein the content ratio of sodium phosphate to biodegradable polymer is about 0.1 to 20% (w / w),

(21) The pharmaceutical composition of (17), wherein the biodegradable polymer is a lactic acid-glycolic acid copolymer,

(22) The pharmaceutical composition of (21) above 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.

(23) The pharmaceutical composition of (1), wherein the biodegradable polymer is an aliphatic polyester,

(24) The pharmaceutical composition of (23), wherein the aliphatic polyester is a lactic acid-glycolic acid copolymer,

(25) the pharmaceutical composition of the above (1) in the form of a suspension,

(26) the pharmaceutical composition of (1), which is used for injection,

(27) the use of the pharmaceutical composition according to the above (1) for the manufacture of a medicament for the treatment or prevention of bone diseases,

(28) the use of biodegradable polymers for enhancing bone regeneration enhancing activity,

(29) A method for treating or preventing a bone disease in a mammal, comprising administering an effective amount of the pharmaceutical composition according to (1) to a subject.

(30) the method according to (29) above, wherein the bone disease is fracture;

(31) An agent for treating or preventing bone diseases with enhanced bone regeneration enhancing activity, which contains a non-peptide bone regeneration-promoting substance and a biodegradable polymer.

Useful non-peptide bone regeneration enhancing agents of the present invention include (2R, 4S)-(-)-N- [4- (diethoxyphosphorylmethyl) phenyl] -1,2 described in US 5,071,841, US 5,158,943, and JP 5294960. Sulfur-containing heterocyclic compounds or salts thereof, such as 4,5-tetrahydro-4-methyl-7,8-methylenedioxy-5-oxo-3-benzothiepin-2-carboxamide, described in EP 625522 Penzopyran derivatives or salts thereof, such as N- (4-dimethoxyphosphorylmethylphenyl) -4-oxo-4H-1-benzopyran-2-carboxamide, diethyl 4- (7) described in WO 96/01267 Phosphonic acid derivatives or salts thereof, such as cyclohexyl-3,4-dihydro-2-naphthalenecarboxamide) benzylphosphonate, Journal of Pharmacology and Experimental Therapeutics, Vol. 258, pp. 1120-1126 (1991) Prostaglandin A ₁ derivative described in the, Bioorganic Medicinal Chemistry Letters, vitamin D ₃ derivative described in Volume 3, pages 1815-1819 (1993), benzylfo described in EP 524023 Sulfonic acid derivatives, bisphosphonic acid described in Bone Vol. 13, pages 249-255 (1992), and vitamin K ₂ derivatives described in Biochemical and Biophysical Research Communications, Vol. 187, pages 814-820 (1992).

The pharmaceutical composition of the present invention may contain one or more of the non-peptide bone regeneration enhancing substances described above as the active ingredient.

In the above-mentioned non-peptide bone regeneration enhancing substances, compounds represented by the following formula (I) or salts thereof are preferably used in the present invention.

A compound of formula (I)

[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, and X is -CH (OH)-or -CO-, k is 0 or 1 and k 'is 0, 1 or 2.

In the general formula (I), substituents of the substituted benzene represented by ring A include a halogen atom, a nitro group, an optionally substituted alkyl group, an optionally substituted hydroxyl group, an optionally substituted thiol group, an optionally substituted amino group, an Practical groups, mono- or di-alkoxyphosphoryl groups, phosphono groups, optionally substituted aryl groups, optionally substituted aralkyl groups, and optionally substituted aromatic heterocyclic groups. One to four, preferably one or two, of the same or different substituents may be present in the benzene ring.

Halogen atoms include fluorine, chlorine, bromine, and iodine.

Alkyl groups of optionally substituted alkyl groups include methyl, ethyl, propyl, isopropyl, butyl, isobutyl, s-butyl, t-butyl, pentyl, isopentyl, neopentyl, hexyl, heptyl, octyl, nonyl and decyl; And cycloalkyl having 3 to 7 carbon atoms such as an alkyl group having 1 to 10 carbon atoms, cyclopropyl, cyclobutyl, cyclohexyl and cycloheptyl. These alkyl groups are halogen atoms (eg fluorine, chlorine, bromine, iodine), hydroxyl groups, alkoxy groups having 1 to 6 carbon atoms (eg methoxy, ethoxy, propoxy, butoxy, hexyloxy), mono or It may also be substituted with 1 to 3 substituents selected from di-C _1-6 alkoxyphosphoryl groups (eg, methoxyphosphoryl, ethoxyphosphoryl, dimethoxyphosphoryl, diethoxyphosphoryl) and phosphono groups.

Substituted alkyl groups include trifluoromethyl, trifluoroethyl, trichloromethyl, hydroxymethyl, 2-hydroxy-ethyl, methoxyethyl, 1-methoxyethyl, 2-methoxyethyl, 2,2 -Diethoxyethyl, 2-diethoxyphosphorylethyl, phosphonomethyl, etc. are mentioned.

As a substituted hydroxyl group, an alkoxy group, an alkenyloxy group, an aralkyloxy group, an acyloxy group, an aryloxy group, etc. are mentioned. Preferred alkoxy is an alkoxy group having 1 to 10 carbon atoms (e.g., methoxy, ethoxy, propoxy, butoxy, t-butoxy, pentyloxy, hexyloxy, heptyloxy, nonyloxy) and 4 to 6 carbon atoms. Cycloalkoxy groups (for example, cyclobutoxy, cyclopentoxy, cyclohexyloxy) are mentioned. Preferred alkenyloxy groups are alkenyloxy groups having 2 to 10 carbon atoms such as allyloxy, crotyloxy, 2-pentenyloxy, 3-hexenyloxy, 2-cyclopentenylmethoxy and 2-cyclohexenylmethoxy Can be mentioned. Preferred aralkyloxy groups are aralkyloxy having 6 to 19 carbon atoms, more preferably C _6-14 aryl-C _1-4 alkyloxy groups (eg benzyloxy, phenethyloxy). Preferred acyloxy groups are alkanoyloxy groups of 2 to 10 carbohydrates (eg, acetyloxy, propionyloxy, n-butyryloxy, hexanoyloxy). Preferred aryloxy groups are aryloxy groups having 6 to 14 carbon atoms (eg, phenoxy, biphenyloxy). In addition, these groups may be substituted by 1 to 3 substituents selected from the aforementioned halogen atoms, hydroxyl groups, alkoxy groups having 1 to 6 carbon atoms, mono- or di-C _1-6 alkoxyphosphoryl groups and the like. Substituted hydroxyl groups include trifluoromethoxy, 2,2,2-trifluoroethoxy, difluoromethoxy, 2-methoxyethoxy, 4-chlorobenzyloxy and 2- (3,4- Dimethoxyphenyl) ethoxy etc. are mentioned.

Substituted thiol groups include alkylthio groups, aralkylthio groups, and acylthio groups. Preferred alkylthio groups are alkylthio groups having 1 to 10 carbon atoms (eg, methylthio, ethylthio, propylthio, butylthio, pentylthio, hexylthio, heptylthio, nonylthio) and cycloalkylthio groups having 4 to 6 carbon atoms ( Cyclobutylthio, cyclopentylthio, cyclohexylthio). Preferred aralkylthio groups are aralkylthio having 7 to 19 carbon atoms, more preferably C _6-14 aryl-C _1-4 alkylthio groups such as benzylthio and phenethylthio. Preferred acylthio groups are alkanoylthio groups having 2 to 10 carbon atoms (eg, acetylthio, propionylthio, n-butyrylthio, hexanoylthio). In addition, these substituted thiol groups may be substituted by 1 to 3 substituents selected from the aforementioned halogen atoms, hydroxyl groups, alkoxy groups having 1 to 6 carbon atoms, mono- or di-C _1-6 alkoxyphosphoryl groups, and the like. . Specifically substituted thiol groups are trifluoromethylthio, 2,2,2-trifluoroethylthio, 2-methoxyethylthio, 4-chlorobenzylthio, 3,4-dichlorobenzylthio, 4-fluoro Benzylthio, 2- (3,4-dimethoxyphenyl) ethylthio, and the like.

As a substituent of the substituted amino group, the above-mentioned alkyl group of 1 to 10 carbon atoms, alkenyl group of 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 ), One or two identical or different substituents selected from aryl groups having 6 to 14 carbon atoms (eg phenyl, naphthyl) and aralkyl groups having 7 to 19 carbon atoms (eg benzyl) may be used.

These substituents may be substituted by the aforementioned halogen atoms, alkoxy groups having 1 to 6 carbon atoms, mono or di-C _1-6 alkoxyphosphoryl groups, phosphono groups and the like. Specifically, substituted amino groups include methylamino, dimethylamino, ethylamino, diethylamino, dibutylamino, diallylamino, cyclohexylamino, phenylamino, N-methyl-N-phenylamino, N-methyl- N- (4-chlorobenzyl) amino, N, N-di (2-methoxyethyl) amino, etc. are mentioned.

As said acyl group, the organic carboxylic acid acyl group and sulfonic acid acyl group which have a C1-C6 hydrocarbon group (for example, methyl, ethyl, n-propyl, hexyl, phenyl) are mentioned. Useful organic carboxylic acyl groups include formyl, C _1-10 alkyl-carbonyl groups (e.g., acetyl, propionyl, butyryl, valeryl, pivaloyl, hexanoyl, octanoyl, cyclobutanecarbonyl, cyclohexancar Carbonyl, cycloheptancarbonyl), C _2-10 alkenyl-carbonyl group (e.g. crotonyl, 2-cyclohexenecarbonyl), C _6-14 aryl-carbonyl group (e.g. benzoyl), C _{7-19 aralkyl-} Carbonyl groups (eg benzylcarbonyl, benzhydrylcarbonyl), 5- or 6-membered aromatic heterocyclic carbonyl groups (eg nicotinoyl, 4-thiazolylcarbonyl) and 5- or 6-membered aromatic heterocyts And click acetyl groups (eg, 3-pyridylacetyl, 4-thiazolylacetyl). Useful sulfoneacyl groups having 1 to 6 carbon atoms include methanesulfonyl and ethanesulfonyl.

These acyl groups may be substituted with 1 to 3 substituents selected from the aforementioned halogen atoms, hydroxyl groups, alkoxy groups having 1 to 6 carbon atoms, amino groups and the like. Specifically, examples of the substituted acyl group include trifluoroacetyl, trichloroacetyl, 4-methoxybutyryl, 3-cyclohexyloxypropionyl, 4-chlorobenzoyl, 3,4-dimethoxybenzoyl, and the like. .

As mono- or di-alkoxyphosphoryl groups, mono such as methoxyphosphoryl, ethoxyphosphoryl, propoxyphosphoryl, isopropoxyphosphoryl, butoxyphosphoryl, pentyloxyphosphoryl and hexyloxyphosphoryl _{Such as} C _1-6 alkoxyphosphoryl groups, dimethoxyphosphoryl, diethoxyphosphoryl, dipropoxyphosphoryl, diisopropoxyphosphoryl, dibutoxyphosphoryl, dipentyloxyphosphoryl and dihexyloxyphosphoryl And di-C _1-6 alkoxyphosphoryl groups, among which dimethoxyphosphoryl, diethoxyphosphoryl, dipropoxyphosphoryl, diisopropoxyphosphoryl, ethylenedioxyphosphoryl, dibutoxyphosphoryl Di-C _1-6 alkoxyphosphoryl groups such as the like are preferable.

Examples of the aryl group of an optionally substituted aryl group include aryl groups having 6 to 14 carbon atoms such as phenyl, naphthyl and anthryl. These aryl groups may be substituted by one to three substituents selected from alkyl, halogen atoms, hydroxyl groups, alkoxy groups having 1 to 6 carbon atoms and the like mentioned above. Specifically, substituted aryl groups include 4-chlorophenyl, 3,4-dimethoxyphenyl, 4-cyclohexylphenyl, and 5,6,7,8-tetrahydro-2-naphthyl.

Aralkyl groups of the optionally substituted aralkyl group include aralkyl groups having 7 to 19 carbon atoms such as benzyl, naphthylethyl and trityl. These aralkyl groups may be substituted on the aromatic ring by one to three substituents selected from the aforementioned alkyl groups of 1 to 10 carbon atoms, halogen atoms, hydroxyl groups, alkoxy groups of 1 to 6 carbon atoms, and the like. Specifically, substituted aralkyl groups include 4-chlorobenzyl, 3,4-dimethoxybenzyl, 4-cyclohexylbenzyl and 5,6,7,8-tetrahydro-2-naphthylethyl.

Aromatic heterocyclic groups of optionally substituted aromatic heterocyclic groups include those having from 1 to 4 atoms of nitrogen, oxygen and / or sulfur such as furyl, thienyl, imidazole, thiazolyl, oxazolyl and thidiazolyl 5-6 membered aromatic heterocyclic group is mentioned. These aromatic heterocyclic groups may be substituted by one to three substituents selected from alkyl, halogen atoms, hydroxyl groups, alkoxy groups having 1 to 6 carbon atoms and the like mentioned above.

If two alkyl groups are present as substituents adjacent to each other on the benzene ring A, they may be bonded together to form an alkylene group represented by the formula-(CH ₂ ) _m where m is an integer from 3 to 5 For example trimethylene, tetramethylene, pentamethylene).

When two alkoxys are present as substituents adjacent to each other on the benzene ring A, they are bonded together to form an alkylenedioxy group represented by the formula -O- (CH ₂ ) _n -O where n is an integer from 1 to 3. It may also form (eg methylenedioxy, ethylenedioxy, trimethylenedioxy).

In these cases, 5 to 7 membered rings are formed with the carbon atoms of the benzene ring.

In formula (I), R is a hydrogen atom or an optionally substituted hydrocarbon group.

The optionally substituted hydrocarbon group represented by R includes the aforementioned alkyl group (preferably an alkyl group having 1 to 10 carbon atoms), an alkylene group (preferably an alkylene group having 2 to 10 carbon atoms), and an aryl group (preferably having 6 to 6 carbon atoms). An aryl group of 14) and an aralkyl group (preferably an aralkyl group having 7 to 19 carbon atoms). Useful substituents on the hydrocarbon group include the aforementioned 5-6 membered aromatic heterocyclic groups, halogen atoms, di-C _1-6 alkoxyphosphoryl groups and phosphono groups.

Preferred examples of R include unsubstituted alkyl groups having 1 to 6 carbon atoms such as methyl, ethyl, propyl, isopropyl, butyl, isobutyl, s-butyl, t-butyl, pentyl, neopentyl and hexyl.

In the formula (I), B is a carboxyl group which is optionally esterified or amidated.

The esterified carboxyl group represented by B is an alkoxycarbonyl group, preferably a C _1-10 alkoxy-carbonyl group (eg methoxycarbonyl, ethoxycarbonyl, propoxycarbonyl, butoxycarbonyl), aryloxy-carbonyl group And preferably C _6-14 aryloxy-carbonyl group (e.g. phenoxycarbonyl) and aralkyloxycarbonyl group, preferably C _7-19 aralkyloxy-carbonyl group (e.g. benzyloxycarbonyl group).

The amidated carboxyl group represented by B is represented by the formula -CON (R ₁ ) (R ₂ ), wherein R ₁ and R ₂ are independently a hydrogen atom, an optionally substituted hydrocarbon group or an optionally substituted 5 to 7 membered hetero group. Optionally substituted carbamoyl groups represented by cyclic group).

As the hydrocarbon group of the optionally substituted hydrocarbon group represented by R ₁ or R ₂ , the above-mentioned alkyl group, preferably an alkyl group having 1 to 10 carbon atoms (eg, methyl, ethyl, propyl, isopropyl, butyl, isobutyl, s -Butyl, t-butyl, pentyl, isopentyl, neopentyl, hexyl, heptyl, octyl, nonyl, decyl), alkenyl groups, preferably alkenyl groups having 2 to 10 carbon atoms (e.g. allyl, vinyl, 2-pentene- 1-yl, 3-penten-1-yl, 2-hexen-1-yl, 3-hexen-1-yl, 2-cyclohexenyl, 2-cyclopentenyl, 2-methyl-2-propene-1 -Yl, 3-methyl-2-buten-1-yl), an aryl group, preferably an aryl group having 6 to 14 carbon atoms (e.g., phenyl, naphthyl, anthryl) and an aralkyl group, preferably having 7 to 7 carbon atoms And 19 aralkyl groups (eg benzyl, naphthyl, trityl). These hydrocarbon groups are halogen atoms (eg fluorine, chlorine, bromine, iodine), hydroxyl groups, alkoxy groups having 1 to 6 carbon atoms (eg methoxy, ethoxy, propoxy, butoxy, t-butoxy, pentyloxy , Hexyloxy), an amino group which may be substituted by an alkyl group having 1 to 6 carbon atoms (eg, amino, methylamino, ethylamino, dimethylamino, diethylamino, dipropylamino), substituted by C _1-10 acyl group Carbamoyl groups (eg, carbamoyl, methylcarbamoyl, dimethylcarbamoyl, diethyl) which may be substituted by substituted amino groups (eg, acetylamino, propionylamino, benzoylamino), alkyl groups having 1 to 6 carbon atoms Carbamoyl), C _1-6 alkoxy-carbonyl groups (eg methoxycarbonyl, ethoxycarbonyl, propoxycarbonyl), mono- or di-alkoxyphosphoryl groups (eg dimethoxyphosphoryl, diethoxy Phosphoryl, ethylenedi Mono or di-C _1-6 alkoxyphosphoryl, such as oxyphosphoryl), mono or di-alkoxyphosphorylalkyl groups (e.g., methoxyphosphorylmethyl, ethoxyphosphorylmethyl, methoxyphosphorylethyl, ethoxyphosph) Mono or di-C _1-6 alkoxyphosphoryl-C _1-3 alkyl groups such as forylethyl, dimethoxyphosphorylmethyl, diethoxyphosphorylmethyl, dimethoxyphosphorylethyl, diethoxyphosphorylethyl), part:

(Wherein p is an integer of 2 to 4), a phosphono group, and 1 to 3 substituents selected from the aforementioned aromatic heterocyclic groups may be substituted.

As the 5-7 membered heterocyclic group of the optionally substituted 5-7 membered heterocyclic group represented by R ₁ or R ₂ , a 5-7 membered heterocyclic group containing sulfur, nitrogen or oxygen atoms, 2 5 or 6 membered heterocyclic groups containing 4 to 4 nitrogen atoms, and 5 or 6 membered heterocyclic groups containing 1 or 2 nitrogen atoms and sulfur or oxygen atoms. These heterocyclic groups may be condensed with six-membered rings containing up to two nitrogen atoms, benzene rings or five-membered rings containing sulfur atoms.

As the substituent of the optionally substituted 5 to 7-membered heterocyclic group represented by R ₁ or R ₂ is, it may have the same substituent substituted hydrocarbon group is used one to four dogs indicated by R ₁ or R _2.

Preferred examples of the 5 to 7 membered heterocyclic group represented by R ₁ or R ₂ include 2-pyridyl, pyrimidyl, pyrazinyl, pyridazinyl, pyrazolyl, imidazolyl, thiazolyl, oxazoline, pyrido [2,3-d] pyrimidyl, benzopyranyl, 1,8-naphthyridyl, quinolyl, thieno [2,3-b] pyridyl, tetrazolyl, thiadiazolyl, oxadiazolyl, triazinyl, Triazolyl, thienyl, pyrrolyl, pyrrolinyl, furyl, pyrrolidinyl, benzothienyl, indolyl, imidazolidinyl, piperidyl, piperidino, piperazinyl, morpholinyl and morpholino Can be mentioned.

Partial-NR ₁ (R ₂ ) may combine with R ₁ and R ₂ to form a 5 to 7 membered ring. Such rings include morpholine, piperidine, thiomorpholine, homopiperidine, piperidine, pyrrolidine, thiazolidine and azepine.

Substituted alkyl groups as preferred examples of the optionally substituted hydrocarbon group represented by R ₁ or R ₂ 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-thi Nil) 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-dimethyl Carbamoylpentyl, ethoxycarbonylmethyl, isopropoxycarbonylethyl, t-butoxycarbonylpropyl, 2-diethoxyphosphorylethyl, 3-dipropoxyphosphorylpropyl, 4-dibutoxyphosphorylbutyl , Ethylenedioxyphosphorylmethyl, 2-phosphonoethyl and 3-foam Phosphonopropyl. Preferably substituted aralkyl groups are 4-chlorobenzyl, 3- (2-fluorophenyl) propyl, 3-methoxybenzyl, 3,4dimethoxyphenethyl, 4-ethylbenzyl, 4- (3-trifluoro Romethylphenyl) butyl, 4-acetylaminobenzyl, 4-dimethylaminophenethyl, 4-diethoxyphosphorylbenzyl and 2- (4-dipropoxyphosphorylmethylphenyl) ethyl. Preferred substituted aryl groups include 4-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-trifluoro Ethoxy) phenyl, 4-propionylphenyl, 4-cyclohexanecarbonylphenyl, 4-dimethylaminophenyl, 4-benzoylaminophenyl, 4-diethoxycarbamoylphenyl, 4-t-butoxycarbonylphenyl, 4-diethoxyphosphorylphenyl, 4-diethoxyphosphorylmethylphenyl, 4- (2-diethoxyphosphorylethyl) phenyl, 2-diethoxyphosphorylmethylphenyl, 3-diethoxyphosphorylmethylphenyl, 4-dipropoxy Phosphorylphenyl, 4- (2-phosphonoethyl) phenyl, 4-phosphonomethylphenyl and 4-phosphonophenyl. Preferably substituted 5-7 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-isooxazolyl, 4- (4-chlorophenyl) -5-methyl-2-oxazolyl, 3-phenyl-1,2,4-thiadiazol-5-yl , 5-methyl-1,3,4-thiadiazol-2-yl, 5-acetylamino-2-pyrimidyl, 3-methyl-2-thienyl, 4,5-dimethyl-2-furanyl and 4 -Methyl-2-morpholinyl is mentioned.

For Formula I, Ring A is preferably selected from ① halogen atoms, ② optionally substituted alkyl groups, ③ optionally substituted hydroxyl groups, ④ optionally substituted thiol groups and / or ⑤ optionally substituted amino groups Benzene rings which may be substituted with one or more, preferably one or two substituents.

More preferably, ring A is a halogen atom mentioned above, an alkyl group having 1 to 10 carbon atoms (more preferably 1 to 5 carbon atoms), an alkoxy group having 1 to 10 carbon atoms (more preferably 1 to 5 carbon atoms), 1 selected from an alkylenedioxy group represented by -O- (CH ₂ ) _n -O (wherein n is an integer of 1 to 3) and / or an alkylthio group having 1 to 10 carbon atoms (more preferably 1 to 5 carbon atoms). Or a benzene ring which may be substituted by two substituents.

More preferably ring A is a benzene ring which may be substituted with an alkylenedioxy group represented by the formula -O- (CH ₂ ) _n -O (wherein n is an integer from 1 to 3).

B is preferably an alkoxy-carbonyl group or a formula -CON (R ₁ ) (R ₂ ) wherein R ₁ and R ₂ are hydrogen atoms, optionally substituted hydrocarbon groups or optionally substituted 5-7 membered heterocyclic It is.

In R ₁ and R ₂ , R ₁ is preferably a hydrogen atom or an alkyl group having 1 to 10 carbon atoms (eg, methyl, ethyl, propyl), and R ₂ is preferably a halogen atom (eg, fluorine, chlorine, Bromine), C _1-6 alkoxy (eg methoxy, ethoxy), mono or di-alkoxyphosphoryl (preferably mono or di-C _1-6 alkoxyphosphoryl, such as diethoxyphosphoryl), mono or Di-alkoxyphosphorylalkyl (preferably mono or di-C _1-6 alkoxyphosphoryl-C _1-3 alkyl, such as diethoxyphosphorylmethyl), or C _1-6 alkoxycarbonyl (eg, methoxycarbon Phenyl or phenyl-C _1-3 alkyl group which may be substituted by carbonyl, ethoxycarbonyl), or 5 or 6 containing 1 or 2 nitrogen atoms or nitrogen and sulfur atoms. Membered heterocyclic group (such as pyridyl).

More preferred examples of R ₁ and R ₂ include those in which R ₁ is a hydrogen atom and R ₂ is phenyl substituted with mono or di-C _1-6 alkoxyphosphoryl-C _1-3 alkyl (eg, 4-diethoxy Phosphorylmethylphenyl).

In formula (I), X is -CH (OH)-or -CO-, preferably -CO-.

In formula I, k is 0 or 1, k 'is 0, 1 or 2, preferably k is 1 and k' is 0.

R is preferably a hydrogen atom, an alkyl group having 1 to 6 carbon atoms (eg, methyl, ethyl) or a phenyl group.

Compound (I) is preferably an optically active compound represented by the following formula II:

[Formula II]

(Wherein R ₃ is a lower alkyl group and R ₄ and R ₅ are independently a lower alkyl group or combine with each other to form a lower alkylene group).

Lower alkyl groups represented by R ₃ , R ₄ and R ₅ in Formula II have carbon numbers such as methyl, ethyl, propyl, isopropyl, butyl, isobutyl, s-butyl, t-butyl, pentyl, isopentyl, neopentyl and hexyl The alkyl group of 1-6 (preferably 1-4) is mentioned. R ₄ and R ₅ may be bonded to each other to form a lower alkylene group. In this case, part: Part: (p is an integer from 2 to 4)

It may also indicate.

Preferred groups of R ₃ , R ₄ and R ₅ include alkyl groups having 1 to 4 carbon atoms such as methyl and ethyl.

Compound II is an optically active compound having a (2R, 4S) configuration and contains substantially no compound having a (2S, 4R) configuration. Preference is given to compounds II having an optical purity of almost 100%.

Most preferably compound II is, for example, (2R, 4S)-(-)-N- [4- (diethoxyphosphorylmethyl) phenyl] -1,2,4,5-tetrahydro-4-methyl-7, 8-methylenedioxy-5-oxo-3-benzothiene-2-carboxamide (hereinafter also referred to as compound A).

Salts of non-peptide bone regeneration enhancing substances of the present invention are preferably pharmaceutically acceptable salts. Pharmaceutically acceptable salts include salts with inorganic bases, salts with organic bases, and salts with basic or acidic amino acids. Inorganic bases capable of forming such salts include alkali metals (eg sodium, potassium) and alkaline earth metals (eg calcium, magnesium), and such organic bases include trimethylamine, triethylamine, pyridine, picoline, N, N-dibenzylethylenediamine and diethanolamine, and such inorganic acids include hydrochloric acid, bromic acid, iodic acid, phosphoric acid, nitric acid and sulfuric acid, and such organic acids include formic acid, acetic acid and trifluoro Roacetic acid, oxalic acid, tartaric acid, fumaric acid, maleic acid, methanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid and citric acid, and such basic or acidic amino acids include arginine, lysine, aspartic acid and glutamic acid.

Non-peptide bone regeneration enhancing substances of the present invention can be prepared, for example, by commonly known methods (e.g., US 5071841, US 5158943 described above) and by the methods described below or by methods of improvement thereof. For example, compound I or a salt thereof may be cyclized to a compound of formula III or a salt thereof and, if necessary, subjected to oxidation and / or hydrolysis, or to hydrolysis and additional amidation, and then to the reaction product. After oxidation, it may be prepared by reduction if necessary.

(Wherein B 'is an esterified carboxyl group, Y is a hydroxy group or a halogen atom and other symbols are as defined above).

For example, compound II is prepared by reacting a reactive derivative or a salt thereof in an optically active compound represented by the following formula (IV) or a carboxyl group thereof with a compound represented by the following formula (V), an amino group thereof or a salt thereof.

Wherein R ₃ is the same as defined above

Wherein R ₄ and R ₅ are the same as defined above.

Preferred reactive derivatives of the amino groups on compound V include Schiff base type imino or enamine form tautomeric isomers resulting from the reaction of compound V with a carbonyl compound such as an aldehyde (eg acetaldehyde) or a ketone (eg acetone); Silyl derivatives resulting from the reaction of compound V with a silyl compound such as bis (trimethylsilyl) acetamide, mono (trimethylsilyl) acetamide or bis (trimethylsilyl) urea; And derivatives resulting from the reaction of compound V with phosphorus trichloride or phosgene.

Preferred reactive derivatives of the carboxyl groups on compound IV are acid halides, acid anhydrides, active amides and active esters obtained by conventional methods. Preferred reactive derivatives include acid chlorides; Acid azide; Substituted phosphoric acids such as dialkylphosphoric acid, phenylphosphoric acid, diphenylphosphoric acid, dibenzyl phosphoric acid or halogenated phosphoric acid, sulfonic acids such as sulfuric acid, thiosulfic acid, sulfuric acid, methanesulfonic acid, acetic acid, propionic acid, butyric acid, isobutyric acid, pi Aliphatic carboxylic acids such as carboxylic acid, petanoic acid, isopentanoic acid or trichloroacetic acid, or mixed acid anhydrides with aromatic carboxylic acids such as benzoic acid; Symmetric acid anhydrides; Imidazole, 4-substituted amidazole, dimethylpyrazole, triazole or tetrazole with activated amide; Cyanomethyl ester, methoxymethyl ester, dimethyliminomethyl ester, vinyl ester, propargyl ester, p-nitrophenyl ester, trichlorophenyl ester, pentachlorophenyl ester, mesylphenyl ester, phenylazophenyl ester, phenyl Active esters such as thio esters, p-nitrophenyl esters, p-clasylthio esters, carboxymethylthio esters, pyranyl esters, pyridyl esters, piperidyl esters, or 8-quinolylthio esters; And N, N-dimethylhydroxylamine, 1-hydroxy-2- (1H) -pyridone, N-hydroxysuccinimide, N-hydroxyphthalimide, 1-hydroxy-1H-benzotriazole And esters with N-hydroxy compounds such as N-hydroxy-5-norobornene-2,3-dicarboxyimide.

These reactive derivatives may be arbitrarily selected depending on the type of compound IV.

Preferred salts of the reactive derivatives of compound IV or V are alkali metal salts such as sodium and potassium salts, alkaline earth metal salts such as calcium and magnesium salts, ammonium salts, and trimethylamine salts, triethylamine salts, pyridine salts, blood Organic base salts such as choline salt, dicyclohexylamine salt and N, N-dibenzylethylenediamine salt.

The reaction is usually carried out with water, alcohols such as methanol, ethanol, acetone, dioxane, acetonitrile, chloroform, methylene chloride, ethylene chloride, tetrahydrofuran, ethyl acetate, N, N-dimethylformamide or pyridine. It is usually carried out in a solvent, but may be carried out in other organic solvents so long as it does not interfere with the reaction. The solvent may be used in admixture with water. In the above reaction, when compound IV or V is used in the form of a free acid or salt thereof, the reaction is selected from N, N'-dicyclohexylcarbodiimide; N-cyclohexyl-N'-morpholinoethylcarbodiimide; N-cyclohexyl-N '-(4-diethylaminocyclohexyl) carbodiimide; N, N'-diethylcarbodiimide; N, N'-diisopropylcarbodiimide; N-ethyl-N '-(3-dimethylaminopropyl) carbodiimide; N, N'-carbonylbis (2-methylimidazole); Pentamethyleneketene-N-cyclohexylimine; Diphenylketene-N-cyclohexylimine; Ethoxyacetylene; 1-alkoxy-1-chloroethylene; Trialkyl phosphites; Ethyl polyphosphate; Isopropyl polyphosphate; Phosphorus oxychloride; Diphenylphosphoryl azide; Thionyl chloride; Oxalyl chloride; Lower alkyl haloformates such as ethyl chloroformmate or isopropyl chloroformmate; Triphenylphosphine; 2-ethyl-7-hydroxybenzisooxazolium salt; 2-ethyl-5- (m-sulfophenyl) isooxazolium hydroxide intermolecular salt; N-hydroxybenzotriazole; 1- (p-chlorobenzenesulfonyloxy) -6-chloro-1H-benzotriazole; Or in the presence of conventional condensing agents, such as Wilsmeier reagent prepared by reacting thionyl chloride, phosgene, trichloromethyl chloroformate or phosphorus oxychloride with N, N'-dimethylformamide. Also preferred are condensing agents such as N, N'-dicyclohexylcarbodiimide in the presence of N-hydroxybenzotriazole or N-hydroxy-5-norbornene-endo-2,3-dicarboxyimide. How to use. The reaction is also carried out in the presence of an inorganic or organic base such as an alkali metal hydrogen carbonate tri (lower) alkylamine, pyridine, N- (lower) -alkylmorpholine or N, N-di (lower) alkylbenzylamine. It may also be performed. The reaction temperature is not limited, but the reaction is usually carried out under heating conditions (-10 to 120 ° C) under cooling. The reaction time is usually about 0.5 to 100 hours, preferably about 1 to 50 hours.

Compound (II) thus obtained may be separated and purified by known separation and purification means such as concentration, concentration under reduced pressure, solvent extraction, crystallization, recrystallization, redissolution and chromatography.

Starting compound IV can be prepared, for example, by optically splitting the racemate of compound IV as disclosed in US 5158943. Specifically, the optically active compound prepares a salt of the racemic of Compound IV and a salt of an optically active base (e.g., optically active α-methylbenzylamine, burucine, quinine, synconin) and determines the solubility difference between the diastereomers obtained. Partial crystallization is repeated on the basis to obtain partially soluble salts in pure form, which are then prepared by performing acid treatment.

Other optically active compounds esterify the racemates of compound IV in optical form based on differences in physical properties between the diastereomers obtained and esterification of optically active alcohols (e.g., optically active methyl lactate, methyl mandelate). It can be prepared by preparing an ester and then performing hydrolysis.

Biodegradable polymers of the present invention are polymers that are soluble or insoluble in water and decompose in vivo. Examples of such polymers include one or more α-hydroxycarboxylic acids (eg, lactic acid, glycolic acid, 2-hydroxybutyric acid, 2-hydroxyvaleric acid, 2-hydroxy-3-methylbutyric acid, 2-hydroxy). Capronic acid, 2-hydroxyisocaproic acid, 2-hydroxycaprylic acid), hydroxydicarboxylic acid (such as malic acid) and hydroxytricarboxylic acid (such as malic acid), lactic acid caprolactone, valero Polymers such as lactones, copolymers and mixtures thereof and derivatives thereof (eg block polymers of polylactic acid, polyglycolic acid and polyethylene glycol), poly-α-cyanoacrylates, poly-β-hydroxybutyric acid, polyalkyl Ethylene oxalate (e.g. polytrimethylene oxalate, polytetramethylene oxalate), polyortho-ester, polyortho- carbonate, polycarbonate (e.g. polyethylene carbonate, polyethylene propylene) Carbonates), polyamino acids (eg, poly-γ-benzyl-L-glutamic acid, poly-L-alanine, poly-γ-methyl-L-glutamic acid), hyaluronate, polystyrene, polymethacrylic acid, acrylic acid-metha Krylic acid copolymer, polyamino acid, dakin stearate, ethyl cellulose, acetyl cellulose, nitrocellulose, maleic anhydride copolymer, ethylene-vinyl acetate copolymer, polyvinyl acetate, polyacrylamide, collagen, gelatin, fibrin and hydroxy Cihatate.

These biodegradable polymers may be in the form of homopolymers, two or more copolymers, or mixtures thereof.

The polymerization reaction may be of random, block or graft type. Preferred biodegradable polymers include aliphatic polyesters.

In view of biodegradability and biocompatibility, polymers and copolymers synthesized from at least one α-hydroxycarboxylic acid are preferred.

Specifically, a copolymer synthesized from one or more types of lactic acid, glycolic acid, 2-hydroxybutyric acid, 2-hydroxyvaleric acid, or the like or a mixture thereof is used.

The biodegradable copolymers of the present invention can be prepared by commonly known methods such as those described in EP 172636 or by improved methods thereof.

The above-mentioned α-hydroxycarboxylic acid may be D-, L- or D, L-configuration, with D, L-configuration being preferred.

Homopolymers of the above-mentioned α-hydroxycarboxylic acids include homopolymers of lactic acid, glycolic acid, and 2-hydroxybutyric acid. Preferred α-hydroxycarboxylic acid is lactic acid. Copolymers of the above-mentioned α-hydroxycarboxylic acid include copolymers of glycolic acid and other α-hydroxycarboxylic acids. Preferred α-hydroxycarboxylic acids are lactic acid and 2-hydroxybutyric acid.

Specifically, useful copolymers include lactic acid-glycolic acid copolymers and 2-hydroxybutyric acid-glycolic acid copolymers, and lactic acid-glycolic acid copolymers are preferred.

The average molecular weight of these biodegradable polymers of the present invention is preferably selected in the range of about 2,000 to 800,000, more preferably about 5,000 to 200,000.

The weight average molecular weight of the lactic acid homopolymer (hereinafter referred to as polylactic acid) is preferably about 5,000 to 100,000, more preferably about 6,000 to 50,000. Polylactic acid can be synthesized, for example, by known production methods described in EP 172636.

The content ratio of lactic acid and glycolic acid in the lactic acid-glycolic acid copolymer is preferably about 100/0 to 50/50 (w / w), 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.

Lactic acid-glycolic acid copolymers can be synthesized, for example, by known production methods described in EP 172636. The copolymer can preferably be synthesized by catalytic glass dehydration polymerization condensation reaction.

In the 2-hydroxybutyric acid-glycolic acid copolymer, the content ratio is preferably about 40 to 70 mol% of glycolic acid, and 2-hydroxybutyric acid corresponds to the remainder. 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. 2-hydroxybutyric acid-glycolic acid copolymers can be synthesized, for example, by known production methods described in EP 172636. The copolymer can preferably be synthesized by catalytic glass dehydration polymerization condensation reaction.

The 2-hydroxybutyric acid-glycolic acid copolymer described above may also be used in admixture with polylactic acid. When the 2-hydroxybutyric acid-glycolic acid copolymer is used in combination 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 context of the present invention, the weight average molecular weight is defined based on polystyrene measured by gel permeation chromatography (GPC). Chloroform was used as a mobile phase, and it measured by GPC column KF804Lx2 (made by Showa Denko) and RI detector L-330 (made by Hitachi).

The amount of biodegradable polymer is variable depending on the pharmacologically active strength of the non-peptide bone regeneration enhancing substance, the rate and time of drug release from the biodegradable polymer, etc. as long as the desired purpose can be achieved. For example, the biodegradable polymer is used in an amount of about 0.2 to 10,000 times (weight ratio), preferably about 1 to 1,000 times, more preferably about 1 to 100 times, based on the amount of bioactive material.

The pharmaceutical composition of the present invention may be prepared by conventional methods for preparing the pharmaceutical composition. For example, it can be prepared by dispersing a non-peptide bone regeneration promoting material in a biodegradable polymer, or by filling a non-peptide bone regeneration promoting material in a preformed hollow biodegradable polymer. Specifically, useful methods include underwater drying, phase separation, spray drying and improved methods thereof.

The form of the pharmaceutical composition of the present invention obtained by the above-mentioned preparation method may be, for example, in the form of fine particles, spheres, rods, needles, pellets, films or creams, but the form is not limited so long as the desired purpose can be achieved.

In the present invention, the pharmaceutical composition of the fine particles is also called microcapsules or microspheres.

Examples of methods for preparing microcapsules are as follows:

(1) underwater drying method (o / w method)

In this method, a solution of an organic solvent containing a biodegradable polymer is first prepared. The organic solvent used to prepare the pharmaceutical composition of the present invention preferably has a boiling point of 120 ° C. or lower. Such organic solvents include halogenated hydrocarbons (eg dichloromethane, chloroform, chloroethane, dichloroethane, trichloroethane, carbon tetrachloride), aliphatic esters (eg ethyl ester, butyl ester), ethers (eg ethyl ether, isopropyl ether) ) And aromatic hydrocarbons (eg, benzene, toluene, xylene).

These solvents may be used in combination of two or more kinds in appropriate proportions. The organic solvent is preferably dichloromethane or acetonitrile, more preferably dichloromethane. The concentration of the biodegradable polymer of the organic solvent solution may vary depending on the molecular weight of the biodegradable polymer and the type of organic solvent, etc., but is usually 0.01 to 80% (w / w), preferably about 0.1 to 70% (w / w), more preferably about 1 to 60% (w / w).

Non-peptide bone regeneration enhancing substances are added to the organic solvent solution containing the biodegradable polymer thus obtained and dissolved and lyophilized or vacuum dried if necessary. The amount of the non-peptide bone regeneration promoting substance may vary depending on the type of drug, the mechanism of action on bone regeneration, the duration of the effect, etc., but it is about 0.001 to 90% (w / w) based on the concentration of the biodegradable polymer of the organic solvent solution. ), Preferably about 0.01 to 80% (w / w), more preferably about 0.1 to 50% (w / w).

The organic solvent solution thus prepared is then added to the water phase to form the o / w emulsion using a turbine type mechanical stirrer or the like. The volume of the aqueous phase is usually selected in the range of about 1 to 10,000 times, preferably about 2 to 5,000 times, more preferably about 5 to 2,000 times the volume of the oil phase.

Emulsifiers may also be added to the aqueous phase. Emulsifiers can be any as long as they can form stable o / w emulsions. Examples of such emulsifiers include anionic emulsifiers, nonionic emulsifiers, polyoxyethylene castor oil derivatives, polyvinylpyrrolidone, polyvinyl alcohol, carboxymethyl cellulose, lecithin, gelatin and hyaluronic acid. These may be used in combination as appropriate. The concentration of the emulsifier in the water phase is preferably about 0.001 to 20% (w / w), more preferably about 0.01 to 10% (w / w), even more preferably about 0.05 to 5% (w / w) to be.

Evaporating the solvent from the oil phase includes evaporating the solvent under normal or slowly reduced pressure during agitation using a propeller stirrer or magnetic stirrer, or evaporating the solvent while adjusting the degree of vacuum using a rotary evaporator or the like. It can be achieved by a method commonly used. The obtained microcapsules are washed several times with, for example, water or heptane to remove free non-peptide bone regeneration enhancing substances, emulsifiers and the like adhering to the surface of the microcapsules and then separated by centrifugation or filtration. After that, the microcapsules are again dispersed in distilled water and then lyophilized.

In the o / w method described above, microcapsules may be prepared by the s / o / w method in which the non-peptide bone regeneration enhancing substance is dispersed in an organic solvent solution containing a biodegradable polymer.

(2) underwater drying method (w / o / w method)

In this method, the non-peptide bone regeneration enhancing substance or salt thereof is first dissolved or dispersed in water to obtain the specific concentration in order to obtain an internal aqueous phase, and if necessary protein (eg gelatin), seaweed (eg agar), Drug-containing substances such as polysaccharides (eg alginic acid), synthetic high molecular weight substances (eg polyvinyl alcohol), basic amino acids (eg arginine, lysine) and the like are added to dissolve or suspend. As a pH adjuster for maintaining the stability and solubility of non-peptide bone regeneration enhancing substances or salts thereof, organic acids such as acetic acid, oxalic acid or citric acid, inorganic acids such as carbonate or phosphoric acid, alkali metal hydroxides such as sodium hydroxide, basic amino acids such as arginine or lysine Or salts thereof (eg, salts with organic acids such as acetic acid, oxalic acid, citric acid, salts with inorganic acids such as carbonic acid, phosphoric acid and hydrochloric acid) may be supplemented to the internal aqueous phase. As stabilizers of non-peptide bone regeneration enhancing substances, proteins (eg albumin, gelatin), starch derivatives (eg dextrin, pullulan), organic acids (eg citric acid), ethylenediaminetetraacetic acid alkali metal salts (eg sodium ethylene Diaminetetraacetic acid salt), hydrogen sulfite alkali metal salt (eg sodium hydrogen sulfite), synthetic high molecular weight material (eg polyethylene glycol) and the like may be added. Common preservatives p-oxybenzoate (eg methyl paraben, propyl paraben), benzyl alcohol, chlorobutanol and thimerosal may also be added.

The amount of the non-peptide bone regeneration enhancing substance may vary depending on the type of drug, the mechanism of action during bone regeneration, or the duration of effect, but is about 0.001 to 90% (w / w), preferably about 0.01 to 80% (w / w), more preferably about 0.1 to 50% (w / w).

The obtained internal water phase is added to a solution containing a biodegradable polymer (oil phase) and emulsified to obtain a w / o emulsion.

Such emulsification is achieved by known dispersion methods including intermittent vibration methods, methods using mixers such as propeller shakers or turbine shakers, colloid mill methods, homogenizer methods and ultrasonic dispersion methods. The solution described above (oil phase) containing the biodegradable polymer is prepared by dissolving the biodegradable polymer in an organic solvent. The solvent may be any solvent as long as it has a boiling point of 120 ° C. or less and is not miscible with water. Such organic solvents include halogenated hydrocarbons (eg dichloromethane, chloroform, chloroethane, dichloroethane, trichloroethane, carbon tetrachloride), aliphatic esters (eg ethyl acetate, butyl acetate), ethers (eg ethyl ether, isopropyl ether) ) And aromatic hydrocarbons (eg, benzene, toluene, xylene). These solvents may be used in combination of two or more kinds in appropriate proportions.

The prepared w / o emulsion is then added to the aqueous phase and the w / o / w emulsion is obtained, followed by evaporation of the oily solvent to obtain microcapsules. The specific procedure for the preparation is the same as that described in (1) above.

(3) Phase separation method

In this method, the dropleting agent is slowly added to the w / o emulsion described above with stirring to precipitate solidify the biodegradable polymer. Droplets can be mixed with solvents of biodegradable polymers, and if the polymer, mineral oil or vegetable oil does not dissolve the solvent for encapsulation, silicone oil, vegetable oil and fat (e.g. sesame oil, soybean oil, corn oil, Cottonseed oil, coconut oil, flaxseed oil), mineral oil, hydrocarbons (eg n-hexane, n-heptane) may be used. These may be used in combination of two or more kinds.

The obtained microcapsules are washed repeatedly with heptane or the like after filtration separation to remove the droplet forming agent. The free drug and the solvent are then removed in the same manner as in water drying. To prevent agglomeration of particles during cleaning, anti-agglomerates: water-soluble sugars such as mannitol, lactol, glucose and starch (eg corn starch), amino acids such as glycine and alanine, proteins such as gelatin, fibrin and collagen may be added. have.

(4) spray drying method

To prepare microcapsules by this method, the above-described w / o emulsion is sprayed through a nozzle into a drying chamber or a spray dryer to evaporate the organic solvent and water in the microdroplets in a short time to obtain microcapsules. The nozzles include dual fluid nozzles, pressure nozzles and rotating disk nozzles. In order to prevent agglomeration of the microcapsules, on the one hand the aqueous solution of the above-mentioned anti-agglomerating agent is sprayed through another nozzle while spraying the w / o emulsion. The microcapsules thus obtained are warmed under reduced pressure to facilitate removal of the water and the solvent contained therein.

In addition to the microcapsules described above, the pharmaceutical compositions of the present invention may be prepared by dispersing a biodegradable polymer in which a non-peptide bone regeneration enhancing substance is dispersed, and forming the sphere, rod, needle, pellet, film, and the like by an appropriate method. Can be prepared. Biodegradable polymers in which non-peptide bone regeneration enhancing substances are dispersed are prepared, for example, according to the method described in US 3773919.

In addition, the pharmaceutical compositions of the present invention may also be prepared by dispersing a biodegradable polymer in which a non-peptide bone regeneration enhancing material is dispersed in an appropriate particle size by the method described in JP 62234656 using a turbo counter jet mill grinder or an ultrasonic jet grinder. Can be. Specifically, the non-peptide bone regeneration enhancing substance is added to and dissolved in an organic solvent containing a biodegradable polymer.

The solid solution obtained by vacuum drying is then roughly ground and then sieved, solvent evaporated, and coarse particles are ground to a constant particle size using an ultrasonic jet mill to obtain a medicament of the present invention.

The pharmaceutical compositions of the present invention may be used in the form of their own microcapsules or may be formulated in various dosage forms with microcapsules, spheres, rods, needles, pellets, films or creams as starting materials. The pharmaceutical compositions of the present invention may contain phosphoric acid or salts thereof (eg sodium phosphate, potassium phosphate) in a proportion of 0 to 30%. Pharmaceutical compositions of the present invention may also be used in parenteral preparations for topical administration (e.g., intramuscular, subcutaneous, organ or connective injections, inwellable preparations, solid preparations such as granules and powders, liquid preparations such as suspensions, ointments May be administered.

Injectable preparations include dispersants (eg, surfactants such as Tween 80 and HCO-60, polysaccharides such as carboxymethyl cellulose, sodium alginate and hyaluronic acid, and polysorbates), preservatives (eg methyl parabens, propyl parabens), isotonic agents (E.g., sodium chloride, mannitol, sorbitol, glucose), buffers (e.g. calcium carbonate), pH adjusters (e.g. sodium phosphate, potassium phosphate) and the like can be prepared in an aqueous suspension by suspending microcapsules in water, and also lecithin It may also be prepared as an oily suspension by dispersing microcapsules in vegetable oils such as sesame oil or corn oil, with or without phospholipids such as fatty acid triglycerides (eg, MIGLYOL 812).

Phosphates may enhance bone regeneration enhancing activity of the pharmaceutical compositions of the present invention.

The concentration of sodium phosphate or potassium phosphate in the injection formulation is about 0.1 mM to 500 mM, preferably about 1 mM to 100 mM.

Preferred embodiments of the present invention are as follows.

(A) lactic acid-glycolic acid copolymer:

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) (2R, 4S)-(-)-N- [4- (diethoxyphosphorylmethyl) phenyl] -1,2,4,5-tetrahydro-4-methyl-7,8-methylenedioxy -5-oxo-3-benzothiine-2-carboxamide, and

(C) sodium phosphate.

The content ratio of (B) to (A) is about 5 to 30% (w / w).

The content ratio of (C) to (A) and (B) is about 0.1 to 20% (w / w).

When microcapsules are used, for example, as injection suspensions, their particle size is selected in the range of average particle diameters of about 0.1 to 300 μm, so long as the requirements for dispersion and needle path are met. Preferably, the particle size is about 1 to 150 μm, more preferably about 2 to 100 μm.

The pharmaceutical composition of the present invention is preferably the suspension described above.

The pharmaceutical composition of the invention is preferably in the form of fine particles. This is because the pharmaceutical composition is less painful to the patient when administered via a needle for conventional subcutaneous or intramuscular injection.

The pharmaceutical composition of the present invention is preferably an injection.

The method of preparing microcapsules as a bactericidal agent includes, but is not limited to, a method of sterilizing the entire manufacturing process, a method of using gamma rays as a bactericide, and a method of adding a preservative.

The pharmaceutical composition of the present invention exhibits sustained release while having improved activity of a non-peptide bone regeneration enhancing substance, and has a sustained period of 1 week to 3 months depending on the type and content of biodegradable polymer, such as a bone disease (eg, fracture). Fractures, osteoporosis, osteomalacia, beset bone syndrome, ankylosing spondylitis, rheumatoid arthritis, osteoarthritis caused by the modification of related arthritis) and related diseases, and prevents bone tissue after surgery such as multiple myelosis, lung cancer and breast cancer. It can be used to regenerate the periodontal tissue in the treatment of root. The pharmaceutical composition of the present invention is particularly effective in fracture patients because the affected part of the patient is fixed and covered with gypsum bandages and it is desired to improve the treatment with a single dose than with multiple doses. Sustained release formulations that make up the pharmaceutical compositions of the invention can be used in combination with other active ingredients. For example, in the case of compounds represented by formula (I) as bone regeneration enhancing substances, examples of active agents to be combined include calcium compounds (eg sodium carbonate), calcitonin, vitamin D (eg alphacalcidol), sex hormones (eg estrogens, Estradiol), prostaglandin A ₁ , bisphosphinic acid, ifriflavone, fluoride compounds (such as sodium fluoride), vitamin K ₂ , BMP (bone morphogenic protein), FGF (fibroblast growth factor), PDGF (platelet derived growth factor) ), TGF-β (modified growth factor-β), IGF-1 (insulin-like growth factor-1), IGF-2 (insulin-like growth factor-2), PTH (parathyroid hormone), and the like.

The pharmaceutical compositions of the present invention with low toxicity can be safely used in mammals (eg, humans, cattle, horses, pigs, dogs, cats, mice, mice, rabbits).

The pharmaceutical composition of the present invention is a high-efficiency, safe drug having a constant drug effect with low toxicity because of the constant release of the drug over a long period of time, preventing and treating bone diseases, repairing damaged bone tissue, and regeneration of the root tissue in the root treatment. It is expected to satisfy such requirements. For example, when the pharmaceutical composition of the present invention is used to treat fractures (eg, femoral neck fractures), it effectively exhibits its enhanced bone regeneration in the local area and can significantly shorten the duration of treatment, typically 2 to 6 months after the start of the fracture. have. Thus, the patient can quickly return to normal life and prevent complications caused by old fractures.

The dosage of the pharmaceutical composition of the present invention depends on the type and content of the non-peptide bone regeneration promoting substance, the release time of the drug and the animal of interest, but may be an effective amount of the non-peptide bone regeneration promoting substance. For example, when the pharmaceutical composition of the present invention is used in the form of microcapsules for treating fracture sites, about 0.01 to 500 mg, preferably based on the active ingredient content (such as Compound I) per adult (50 kg body weight) per administration Preferably about 5-50 mg once weekly to every three months.

Brief description of the drawings

1 is a graph showing the fibula bone mineral content (mg) 2 weeks after the administration of a microcapsule containing a compound A and a placebo microcapsule (control) to mice.

FIG. 2 is a graphical representation of the transient change in compound A retention at the site of administration of the subcapsulated microcapsules containing compound A in the rat back.

The abscissa represents the time after administration (weeks), and the ordinate represents the subcutaneous residual ratio (%) of Compound A.

Figure 3 graphically shows the transient concentration change of Compound A in rat blood in the subcutaneous transplant of microcapsules containing Compound A in the back of rats.

The abscissa represents the time (weeks) after administration, and the ordinate represents the concentration (μg / ml) of Compound A in the blood.

Best Mode for Carrying Out the Invention

The present invention will be explained in more detail by the following reference examples, working examples and test examples, which are not limiting. In the following working examples and test examples, room temperature is defined as a temperature of about 0 to 30 ° C.

Example

Reference Example 1

(2R, 4S)-(-)-1,2,4,5-tetrahydro-4-methyl-7,8-methylenedioxy-5-oxo-3-benzothiene-2-carboxylic acid (R Preparation of) -α-methoxycarbonylbenzyl ester.

A solution of 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide hydrochloride (12.59 g) in dichloromethane (200 ml) was added dropwise with N, N-dimethylformamide (DMF) (200 ml) at 0 ° C. (±) -t-1,2,4,5-tetrahydro-4-methyl-7,8-methylenedioxy-5-oxo-3-benzothiene-2-carboxylic acid (15.34 g) in Methyl (R)-(-)-mandelic acid (18.19 g) is added followed by 4-dimethylaminopyridine (DMAP) (3.34 g). After stirring for 1 hour at 0 ° C. and 15 hours at room temperature, the mixture is poured into water and extracted with ethyl acetate. The ethyl acetate layer is washed with water and dried over (MgSO ₄ ) and the solvent is evaporated off. The remaining crystals are collected by filtration, washed with ether-hexane and then recrystallized twice with ethyl acetate-hexane to give the title compound (4.09 g yield 17%).

Melting point: 140-141 ℃

Optical rotation [α] _D (23 ° C.): −244.2 ° (c = 0.50, CHCl ₃ )

Reference Example 2

Preparation of (2R, 4S)-(-)-1,2,4,5-tetrahydro-4-methyl-7,8-methylenedioxy-5-oxo-3-benzothiene-2-carboxylic acid .

(2R, 4S)-(-)-1,2,4,5-tetrahydro-4-methyl-7,8-methylenedioxy-5-oxo-3-benzothiene-2 obtained in Reference Example 1 A mixture of carboxylic acid (R) -α-methoxycarbonylbenzyl ester (4.18 g), acetic acid (45 ml) and concentrated hydrochloric acid (30 ml) is stirred for 30 minutes under reflux conditions. The reaction mixture is poured into water (800 ml). The obtained crystals are collected by filtration and dissolved in ethyl acetate (150 ml). The ethyl acetate layer is washed with water and dried over (MgSO ₄ ) and the solvent is evaporated off. The remaining crystals are collected by filtration, washed with hexane and then recrystallized with ethyl acetate-hexane to give the title compound (1.62 g yield 59%) as colorless needles.

Melting Point: 194-195 ℃

Optical rotation [α] _D (23 ° C.): -210.8 ° (c = 0.50, CH ₃ OH)

Reference Example 3

(2R, 4S)-(-)-N- [4- (diethoxyphosphorylmethyl) phenyl] -1,2,4,5-tetrahydro-4-methyl-7,8-methylenedioxy-5- Preparation of oxo-3-benzothiepine-2-carboxamide (Compound A).

A solution of 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide hydrochloride (0.39 g) in dichloromethane (7 ml) was added dropwise at 0 ° C. with N, N-dimethylformamide (DMF) (7 ml). (2R, 4S)-(-)-1,2,4,5-tetrahydro-4-methyl-7,8-methylenedioxy-5-oxo-3-benzothiene- obtained in Reference Example 2 of the To a solution of 2-carboxylic acid (0.47 g) and diethyl 4-aminobenzylphosphonate (0.41 g) is added followed by 1-hydroxybenzotriazole (HOBt) (0.28 g).

After stirring for 1 hour at 0 ° C. and 15 hours at room temperature (25 ° C.), the mixture is poured into water and extracted with ethyl acetate. The ethyl acetate layer is washed with water and dried over (MgSO ₄ ) and the solvent is evaporated off. The remaining crystals are collected by filtration and recrystallized with ethyl-hexane acetate and methanol-hexane to give Compound A (0.37 g yield 44%) in the form of a colorless prism.

Melting Point: 181-182 ℃

Optical rotation [α] _D (23 ° C.): −187.4 ° (c = 0.50, CHCl ₃ )

Example 1

Lactic acid-glycolic acid copolymer (hereinafter referred to as PLGA) (lactic acid-glycolic acid content ratio (mol%) and average molecular weight based on GPC measurement are shown in Table 1) (manufactured by Wako Pure Chemical Industry) and lactic acid homopolymer ( A dichloromethane solution (hereinafter referred to as Solution A) of PLA) is prepared using the compounding ratio shown in Table 1.

Similarly, the dichloromethane solution of compound A (hereinafter referred to as solution B) is prepared using the combination ratios shown in Table 1. Mix solution A and B together uniformly. The mixture was poured into a 0.1% polyvinyl alcohol solution (EG-40, manufactured by Nippon Synthetic Chemicals Co., Ltd.) (hereinafter referred to as PVA solution) at a temperature of 15 ° C. in advance in a specific volume shown in Table 1, and a turbine homomixer at 7000 rpm The emulsion was prepared using to prepare an o / w emulsion, which was then stirred at room temperature for 3 hours to evaporate dichloromethane. After solidification, the oil phase is centrifuged at 2,000 rpm using a principle separator (05PR-22, Hitachi). The collected microcapsule portion is again dispersed in distilled water, then centrifuged and the released drug is washed. The collected microcapsules are again dispersed in a small amount of distilled water and then lyophilized. Microcapsules having the particle sizes indicated in Table 1 were obtained. The ratio of microcapsules included in compound A is 100%.

Microcapsules Solution A Solution B PVA liquid volume (ml) Particle size (㎛) polymer L / G content ratio (mol%) * Weight average molecular weight Polymer weight (g) Dichloromethane (ml) Compound A Weight (g) Dichloromethane (ml) Number 1 PLGA 75/25 10500 2.4 2.5 0.1 1.0 400 24 Number 2 PLGA 75/25 6500 2.4 1.0 0.1 1.0 400 50 Number three PLGA 75/25 17100 2.4 1.5 0.1 1.0 400 38 Number 4 PLA 100/0 12000 2.4 2.0 0.1 1.0 800 37 Number five PLGA 90/10 20000 2.4 2.0 0.1 1.0 400 65 Number 6 PLGA 85/15 12100 2.4 2.0 0.1 1.0 800 51 * Lactic acid / glycolic acid content ratio

Example 2

About 8 g of lactic acid-valerolactone copolymer (manufactured by Taki Chemical, PLV 2500ML, hereinafter referred to as PLV) or glycolic acid-caprolactone copolymer (manufactured by Taki Chemical, PGC 2500MG, hereinafter referred to as PGC) is placed in a centrifuge tube. And heat to about 50 ° C. in a water bath. Approximately 80 mg of Compound A is mixed into each tube and then uniformly dispersed to form an ointment and refrigerated.

Example 3

500 mg of Microcapsule No. 1 obtained in Example 1 are uniformly dispersed in two test tubes of fibrinogen solution for Tisseel (Nippon Zoki Pharmaceutical). Slowly add two test tubes of thrombin solution for Tisseel. Thereafter, each mixture is immediately sucked out with a plastic syringe. The syringe is held at 37 ° C. for 30 minutes to solidify the contents. After solidification, the contents are extruded with a syringe tip and cut into pellets of volume 200 μl using a razor blade.

Example 4

Four milligrams of Compound A are filled into bone defect filler hydroxyapatite (Boneceram P, Sumitomo Pharmaceuticals, 3 mm in diameter, 14 mm in length, pore size of 1 mm). Seal both ends of the hollow with clay.

Example 5

To microcapsule No. 1 of Example 1 containing Compound A (4%), 20% ground gelatin (manufactured by Nitta Gelatin) was added to contain microcapsules weighing 5.5 mm and weighing 125 mg. Obtain a tablet formulation.

Example 6

Compound A (containing 10% content) in the same manner as in Example 1 except that the LGA-glycolic acid content ratio of PLGA (manufactured by Wako Pure Chemical Industry) is 85/15 (mol%) and the weight average molecular weight is 14,900 Prepare microcapsule number 7. The average particle size is 31 μm.

Example 7

A dichloromethane solution containing 2.4 g PLGA (manufactured by Wako Pure Chemical Industry) with a lactic acid-glycolic acid content ratio of 85/15 (mol%) and a weight average molecular weight of 14,900 and 0.1 g of Compound A was prepared in the same manner as in Example 1. And then 0.2 g of estradiol is added. In addition, a PVA solution was added to the solution to prepare an O / W emulsion. Microcapsules No. 8 containing Compound A and estradiol are prepared. The average particle size is 27 μm.

Test Example 1

500 mg of Microcapsule No. 1 obtained in Example 1 were accurately weighed and transferred to a glass centrifuge tube and maintained at 37 ° C. in powder form. After a certain time, the microcapsules are dissolved in a small amount of acetonitrile and then quantitated by high performance liquid chromatography (hereinafter referred to as HPLC). The stability test is shown in Table 2.

Drug content was above 95% even after 4 weeks.

Date Residual rate (%) 0 99 7 97 14 97 21 95 28 96

Test Example 2

The microcapsules Nos. 2, 3 and 4 obtained in Example 1 were weighed exactly 5 mg each and transferred to a glass bottle, followed by 10.0 ml of release test solution (phosphate buffer supplemented with 10% bovine serum albumin, pH 7.0). Shake in a 37 ° C. water bath in the presence (TAITEC, incubator M-100w 115 round trip / min). Take 100 μL of each sample over time and add 100 μL of acetonitrile. After shaking, the mixture is centrifuged. The obtained supernatant is assayed by HPLC to determine the amount of Compound A released. The release test is shown in Table 3.

Microcapsules of three release patterns were obtained.

Work Residual rate (%) Number 2 Number three Number 4 0.0 0.0 0.0 0.0 0.3 81.7 8.4 5.7 1.0 92.2 18.1 10.5 2.0 91.0 27.3 14.8 3.0 88.6 36.6 - 7.0 86.4 67.6 33.3 9.0 82.9 70.8 - 11.0 82.8 74.1 -

Test Example 3

Microcapsule Nos. 3 to 6 obtained in Example 1 were dispersed in 0.3 ml of a dispersant (solution of 1.5 mg carboxymethyl cellulose in distilled water, 0.3 mg of polysorbate 20 and 15 mg of mannitol) at 25 mg each, and 22G. A needle is used to inject subcutaneously into the head of a 5 week old male SD rat (n = 5) under ether anesthesia. Mice are killed at regular intervals after administration. Microcapsules remaining at the site of administration were taken, assayed by HPLC and the amount of Compound A in the microcapsules was measured.

The results are shown in Table 4.

Various microcapsules of different release patterns were likewise obtained in vivo. Specifically, the microcapsule number 3 had one month, the microcapsule numbers 4 and 5 had one month or more, and the microcapsule number 6 had a sustained period of three weeks.

Work Residual rate (%) Number three Number 4 Number five Number 6 0 100 100 100 100 One 89 82 62 76 7 47 70 62 47 14 20 48 55 23 21 17 33 44 2 28 2 17 32 6

Test Example 4

The ointment preparation obtained in Example 2 was filled into an ointment tube. Clips for micropipette attachment were attached to the end of the tube to facilitate administration to animals. Ointment was administered to the back of 5 week old male SD rats (n = 5) under ether anesthesia. Dosage was determined based on tube weight difference before and after administration. The mice are killed at regular time intervals. The amount of residual Compound A was determined by HPLC. The results are shown in Table 5.

Ointments were obtained that released the full dose in 1-2 weeks.

Work Residual rate (%) PGC PLV 0 100 100 One 36 10 7 9 0 14 One One

Test Example 5

Under pentobarbital anesthesia, the heads of 6-week-old SD rats (n = 8) are incised and the periosteum is detached. A 4 mm diameter hole is made in the left cranial canal with a dental drill. One week after suture, 25 mg of Microcapsule No. 3 obtained in Example 1 was dispersed in 0.3 ml of a dispersant (1.5 mg carboxymethyl cellulose in distilled water, 0.3 mg of polysorbate 20 and 15 mg of mannitol), Subcutaneously inject into the right temple using a 22G needle. For control, mice administered 0.3 ml of the dispersant alone were used. Three weeks later the mice were killed. The skull is excised and subjected to soft X-ray analysis. Using the obtained photograph, the bone portion newly formed at the bone defect site is measured by image analysis.

It was shown in the microcapsule-administered group that the newly formed bone fraction was significantly increased, demonstrating the excellent bone regeneration promoting activity of the pharmaceutical composition of the present invention.

Test Example 6

Under pentobarbital anesthesia, the left leg of the 6-week-old SD rat (n = 8) is dissected and the center of the left fibula is incised exposed using a cutter. 25 mg of Microcapsule No. 1 obtained in Example 1 is buried in the nasal bone cutting site and sutured. Two weeks later, the rats are killed. The fibula is dissected and the bone mineral content is measured using a bone mineral analyzer (DSC-600, Aloca Co., Ltd, Tokyo). The bone mineral content of the untreated left fibula was also measured. The bone mineral content in the bone is determined by subtracting the measurement of the left fibula from the measurement of the right fibula. For control, placebo microcapsules without Compound A are prepared in the manner used to prepare microcapsule number 1 and compared with the counterpart in the same manner as in Test Example 5. The result is shown in FIG.

Administration of drug containing microcapsules significantly increases bone mineral content, demonstrating good bone regeneration enhancing activity of the pharmaceutical compositions of the present invention.

Test Example 7

25 mg of Microcapsule No. 1 containing Compound A (content 4%) obtained in Example 1 is locally administered to a fibula fracture model rat by the method described in Test Example 6. After 2-3 weeks, the bone mineral content is measured. For control, bone mineral content of placebo microcapsule (drug-free microcapsules) and spontaneous healing groups is measured. The results are shown in Table 6.

group Bone Mineral Increase (mg) 2 weeks 3 weeks Untreated 9.1 ± 1.2 7.7 ± 1.4 Placebo Microcapsules 10.2 ± 1.0 9.8 ± 1.7 Drug-containing microcapsules 15.2 ± 1.4 15.1 ± 2.2

The drug containing microcapsules group has a significantly higher bone mineral content compared to the control, demonstrating the excellent bone regeneration promoting activity of the pharmaceutical composition of the present invention.

Test Example 8

The microcapsule-containing tablet formulation obtained in Example 5 is implanted subcutaneously in the back of 5 week old male SD rats (n = 8) under ether anesthesia. The content of drug remaining at the site of administration is measured by HPLC over time. The result is shown in FIG. Subject tablets showed six weeks of sustained release properties.

Test Example 9

According to Shinpei Miyamoto, Hideki Yoshikawa, Kunio Takaoka, Goal 7, 85-96 (1983), the tibia of the rabbit (weight 3 to 4 kg) is cut and a goal of 5 mm in length After the defects are formed, the microcapsule-containing tablet formulation obtained in Example 5 is inserted and filled, and the wound site is externally fixed. For control, placebo microcapsules containing tablet formulations that do not contain Compound A are used. Soft X-rays revealed bone attachment in rats receiving a tablet containing Compound A two months after surgery, but no bone attachment in rabbits receiving placebo tablets.

Test Example 10

5 week old male SD rats under ether anesthesia by the method described in Test Example 3 (100 mg / kg for Compound A) containing Compound A (compound 10%) obtained in Example 6 It is administered subcutaneously in the back of (n = 8). Blood drug concentrations are measured by HPLC over time. The result is shown in FIG. After 4 weeks of administration, blood drug concentrations of 0.05 to 0.1 μg / ml were obtained.

Test Example 11

A rat fibula fracture model was prepared by the method described in Test Example 6, and Microcapsule No. 7 containing Compound A (content of 10%) obtained in Example 6 was subjected to lyophilized powder (1 mg / rat) or test. Topical administration is in the form of a suspension of the dispersant described in Example 5 (5 mg / 0.25 ml / rat).

Two weeks later, the bone mineral content is measured by the method described in Test Example 6 and compared to the control placebo microcapsule group.

The results are shown in Table 7.

group Bone Mineral Increase (mg) Freeze dried powder Suspension Placebo microcapsules 8.3 ± 0.7 7.6 ± 0.9 Drug-containing microcapsules 12.2 ± 0.8 10.2 ± 0.8

The drug form is a lyophilized powder or suspension, or the drug-containing microcapsules group has a significant increase in bone mineral content compared to the control group, demonstrating good bone regeneration enhancing activity of the pharmaceutical composition of the present invention.

Test Example 12

A rat fibula fracture model was prepared by the method described in Test Example 6. Various concentrations of sodium phosphate prepared by adjusting the microcapsule No. 7 in distilled water with a solution of D-sorbitol (2.5 g), sodium chloride (0.9 g), polysorbate (0.1 g) and carboxymethyl cellulose (0.5 g) It is suspended in a dispersion medium containing.

The suspension is topically administered (5 mg / 0.25 ml / rat).

After 2 weeks, the bone mineral content is measured by the same method described in Test Example 6.

The results are shown in Table 8.

Sodium Phosphate Content (mM) Bone mineral increase (mg) 0 7.0 ± 0.6 2 9.2 ± 1.4 20 9.6 ± 0.5 100 12.0 ± 1.6

Effects of the Invention

Pharmaceutical compositions of the present invention showing enhanced bone regeneration-promoting activity with low toxicity over long periods of time can be used in various bone diseases of mammals such as fractures, fractures, osteoporosis, osteomalacia, Besett's bone syndrome, ankylosing spondylitis, rheumatoid arthritis, arthritis and related diseases As a prophylactic / treatment agent for the destruction of joint tissue caused by the modification of, it can be safely used as a therapeutic agent for bone tissue after surgery such as multiple myelopathy, lung cancer, breast cancer, and as a rejuvenation promoter in the treatment of root root.

Pharmaceutical compositions containing non-peptide bone regeneration enhancing substances and biodegradable polymers are useful as agents for the treatment and / or prevention of bone diseases.

Claims (31)

A pharmaceutical composition comprising a non-peptide bone regeneration enhancing substance and a biodegradable polymer. The pharmaceutical composition according to claim 1, which further contains phosphoric acid or a salt thereof. The pharmaceutical composition according to claim 1, wherein the non-peptide bone regeneration enhancing substance is a nonsteroidal system. The pharmaceutical composition of claim 1, which is used for topical administration. The pharmaceutical composition of claim 1, which is used to improve fracture treatment. The pharmaceutical composition according to claim 1, which is a sustained release formulation. The pharmaceutical composition according to claim 1, wherein the non-peptide bone regeneration-promoting substance is a compound represented by the following formula (I) or a salt thereof: [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, and X is -CH (OH)-or -CO-, k is 0 or 1 and k 'is 0, 1 or 2. 8. The compound of claim 7, wherein ring A is halogen, C _1-10 alkyl, C _1-10 alkoxy, -O-CH ₂ ) _n -O-, wherein n is 1 to 3 and C _1-10 alkyl Pharmaceutical composition, characterized in that the benzene ring which may be substituted with one or two substituents selected from thio. 8. A compound according to claim 7, wherein B is -CON (R ₁ ) (R ₂ ), wherein R ₁ and R ₂ are independently a hydrogen atom, an optionally substituted hydrocarbon group or an optionally substituted 5-7 membered heterocy. Pharmaceutical composition, characterized in that. 10. The compound of claim 9, wherein R ₁ is a hydrogen atom or a C _1-10 alkyl group, and R ₂ is (i) halogen, C _1-6 alkoxy, mono- or di-C _1-6 alkoxyphosphoryl, mono- or di -C _1-6 alkoxyphosphoryl-C _1-3 alkyl, Wherein p is 2 to 4 or a phenyl or phenyl-C _1-3 alkyl group substituted by C _1-6 alkoxycarbonyl, or (ii) one or two nitrogen atoms or 1 substituted by phenyl Pharmaceutical composition, characterized in that it is a 5- or 6-membered heterocyclic group containing two nitrogen atoms and one sulfur atom. 8. A pharmaceutical composition according to claim 7, wherein R is a hydrogen atom, a C _1-6 alkyl group or a phenyl group. 8. A pharmaceutical composition according to claim 7, wherein k is 1 and k 'is 0. The pharmaceutical composition according to claim 1, wherein the non-peptide bone regeneration enhancing substance is an optically active compound represented by Formula II: [Formula II] (Wherein R ₃ is a lower alkyl group and R ₄ and R ₅ are independently lower alkyl groups or combine with each other to form a lower alkylene group) 14. A pharmaceutical composition according to claim 13 wherein R ₃ , R ₄ and R ₅ are independently C _1-6 alkyl groups. The compound of claim 1, wherein the compound is (2R, 4S)-(-)-N- [4- (diethoxyphosphorylmethyl) phenyl] -1,2,4,5-tetrahydro-4-methyl-7 , 8-methylenedioxy-5-oxo-3-benzothiene-2-carboxamide. The pharmaceutical composition of claim 1, wherein the weight ratio of the biodegradable polymer to the non-peptide bone regeneration enhancing material is about 1 to 100 times. The compound according to claim 1, wherein (2R, 4S)-(-)-N- [4- (diethoxyphosphorylmethyl) phenyl] -1,2,4,5-tetrahydro-4-methyl-7,8- A pharmaceutical composition comprising methylenedioxy-5-oxo-3-benzothiepin-2-carboxamide and a biodegradable polymer. 18. The pharmaceutical composition of claim 17, further comprising phosphoric acid or a salt thereof. 19. The pharmaceutical composition of claim 18, wherein the phosphoric acid or salt thereof is sodium phosphate. 18. (2R, 4S)-(-)-N- [4- (diethoxyphosphorylmethyl) phenyl] -1,2,4,5-tetrahydro-4-methyl for a biodegradable polymer. The content ratio of -7,8-methylenedioxy-5-oxo-3-benzothiefine-2-carboxamide is about 5 to 30% (w / w), and (2R, 4S)-(-)-N -[4- (diethoxyphosphorylmethyl) phenyl] -1,2,4,5-tetrahydro-4-methyl-7,8-methylenedioxy-5-oxo-3-benzothiene-2-car Pharmaceutical composition, characterized in that the content ratio of sodium phosphate to the radiamide and the biodegradable polymer is about 0.1 to 20% (w / w). 18. The pharmaceutical composition of claim 17, wherein the biodegradable polymer is a lactic acid-glycolic acid copolymer. The pharmaceutical composition of claim 21, wherein the ratio of lactic acid / glycolic acid is about 90/10 to 50/50 (w / w) and a weight average molecular weight is about 8000 to 50000. The pharmaceutical composition of claim 1 wherein the biodegradable polymer is an aliphatic polyester. A pharmaceutical composition according to claim 23 wherein the aliphatic polyester is a lactic acid-glycolic acid copolymer. A pharmaceutical composition according to claim 1 in the form of a suspension. The pharmaceutical composition of claim 1, which is used for injection. Use of a pharmaceutical composition according to claim 1 for the manufacture of a medicament for the treatment or prevention of bone diseases. Use of biodegradable polymers to enhance bone regeneration promoting activity. A method of treating or preventing bone disease in a mammal comprising administering to the subject a effective amount of the pharmaceutical composition of claim 1. 30. The method of claim 29, wherein the bone disease is fracture. A non-peptide bone regeneration promoting agent and a biodegradable polymer containing a bone disease treatment or prevention agent with improved bone regeneration promoting activity.