RU2188011C2 - Osteogenesis-activating pharmaceutical composition - Google Patents

Abstract

FIELD: pharmacology. SUBSTANCE: pharmaceutical composition contains nonpeptide osteogenesis- activating optically active substance and copolymer of lactic and glycolic acids to be safely used as a more efficient prophylactic therapeutic preparation for different osseous diseases (for example, osseous fractures). EFFECT: higher efficiency. 10 cl, 3 dwg, 7 tbl

Description

 The invention relates to a pharmaceutical composition having increased osteogenesis activating osteogenic activity, comprising a non-peptide osteogenic activating optically active substance and a biodegradable copolymer, which is an agent useful in the treatment and / or prevention of bone diseases.

BACKGROUND OF THE INVENTION
Bone diseases (for example, bone fractures) can occur in all categories of people due to various causes in sports and in road traffic accidents. And, since it takes a long time to cure them, bone diseases introduce significant difficulties into the patient's daily normal life. In recent years, the number of patients with osteoporosis has increased due to an aging population. So, the frequency of bone fractures of the extremities associated with osteoporosis has increased markedly in proportion to this. In particular, a fracture of the femoral neck makes long-term hospitalization necessary and often leads to internal complications, including dementia due to long-term hospitalization, and creates great social and economic problems. An essential task is to enable early discharge of patients with bone fracture from the hospital.

 Bone fracture healing is a form of wound healing characterized by local manifestation and progression. Typically, various local factors work well at the fracture site to activate in vivo healing. Such factors include peptide-type bioactive substances, such as bone morphogenetic proteins (BMP) and transforming growth factors (TGF), which have been reported to activate osteogenesis in animal models "Proceedings of the National Acagemy of Sciences, USA, vol. 87 , pp. 2220-2224 (1990), and Endocrinology, vol. 124, pp. 2991-2993 (1989). "

As for the non-peptide osteogenic activating substance, for example, prostaglandin A ₁ derivatives, vitamin D ₃ derivatives, benzylphosphonic acid derivatives, phenolsulfophthalenic acid derivatives have been reported.

 The above peptide-type bioactive substances are peptides or proteins with molecular weights of more than 5000 and they are rapidly metabolized in vivo and are not stable. With this in mind, some drugs were obtained in an attempt to obtain satisfactory stability, but all of them did not achieve sufficient osteogenesis-activating activity, and there are no drugs that are satisfactory in terms of quality, etc. "Clinical Orthopedics and Related Research, vol. 178, pp. 274-285." Also, the aforementioned non-peptidic osteogenesis activating substances are not clinically effective in activating osteogenesis of activity for treating bone fractures.

 For these reasons, there is a great need for a high-quality agent for the treatment of bone disease, which is highly stable, safe and active, clinically effective in the long-term treatment of bone fractures.

Description of the invention
An extensive study was carried out to solve these problems and it was found that an agent for treating bone disease containing a non-peptide osteogenesis-activating optically active substance and a biodegradable copolymer can unexpectedly serve to activate bone fractures with stronger osteogenic stimulating activity of a non-peptide osteogenic activating substance than when administered separately. Further research was conducted based on this finding and the present invention was created.

где R₃ представляет низшую алкильную группу; и R₄ и R₅ независимо представляют низшую алкильную группу или связаны вместе с образованием низшей алкиленовой группы и сополимер молочной кислоты и гликолевой кислоты,
(2) фармацевтической композиции по п.(1), в которой R₃, R₄ и R₅ независимо представляют C_1-6-алкильную группу,
(3) фармацевтической композиции по п.(1), в которой соединение представляет собой (2R, 4S)-(-)-N-[4-(диэтоксифосфорилметил)фенил]-1,2,4,5-тетрагидро-4-метил-7,8-метилендиокси-5-оксо-3-бензотиепин-2-карбоксамид,
(4) фармацевтической композиции по п.(1), в которой весовое отношение указанного сополимера относительно непептидного остеогенного активирующего оптически активного вещества является приблизительно 1-100-кратным,
(5) фармацевтической композиции по п.(2), которая дополнительно содержит фосфорную кислоту или ее соль,
(6) фармацевтической композиции по п.(5), в которой фосфорная кислота или ее соль представляет собой фосфат натрия,
(7) фармацевтической композиции по п.(2), в которой относительное содержание (2R, 4S)-(-)-N-[4-(диэтоксифосфорилметил)фенил] -1,2,4,5-тетрагидро-4-метил-7,8-метилендиокси-5-оксо-3-бензотиепин-2-карбоксамида в расчете на указанный сополимер равно приблизительно 5-30% (масс.) и относительное содержание фосфата натрия в расчете на (2R,4S)-(-)-N-[4-(диэтоксифосфорилметил)фенил] -1,2,4,5-тетрагидро-4-метил-7,8-метилендиокси-5-оксо-3-бензотиепин-2-карбоксамид и данный сополимер равно приблизительно 0,1-20% (масс.).Thus, this invention relates to:
(1) a pharmaceutical composition comprising a non-peptide osteogenesis activating optically active substance of the formula:

where R ₃ represents a lower alkyl group; and R ₄ and R ₅ independently represent a lower alkyl group or are linked together to form a lower alkylene group and a copolymer of lactic acid and glycolic acid,
(2) the pharmaceutical composition according to (1), in which R ₃ , R ₄ and R ₅ independently represent a C _1-6 alkyl group,
(3) the pharmaceutical composition according to (1), wherein the compound is (2R, 4S) - (-) - N- [4- (diethoxyphosphorylmethyl) phenyl] -1,2,4,5-tetrahydro-4- methyl 7,8-methylenedioxy-5-oxo-3-benzothiepin-2-carboxamide,
(4) the pharmaceutical composition according to (1), wherein the weight ratio of said copolymer relative to the non-peptide osteogenic activating optically active substance is about 1-100-fold,
(5) the pharmaceutical composition according to (2), which further comprises phosphoric acid or a salt thereof,
(6) the pharmaceutical composition according to (5), wherein the phosphoric acid or its salt is sodium phosphate,
(7) the pharmaceutical composition according to (2), wherein the relative content of (2R, 4S) - (-) - N- [4- (diethoxyphosphorylmethyl) phenyl] -1,2,4,5-tetrahydro-4-methyl -7,8-methylenedioxy-5-oxo-3-benzothiepine-2-carboxamide based on the indicated copolymer is approximately 5-30% (mass.) And the relative content of sodium phosphate based on (2R, 4S) - (-) -N- [4- (diethoxyphosphorylmethyl) phenyl] -1,2,4,5-tetrahydro-4-methyl-7,8-methylenedioxy-5-oxo-3-benzothiepin-2-carboxamide and this copolymer is approximately 0, 1-20% (mass.).

(8) the pharmaceutical composition according to (1), wherein the lactic acid / glycolic acid ratio is approximately 90 / 10-50 / 50 wt.% And the weight average molecular weight is approximately 8000-50000,
(9) the pharmaceutical composition according to (1) in the form of a suspension,
(10) the pharmaceutical composition according to (1) used for injection, etc.

Useful non-peptide osteogenic activating optically active substances of the present invention include sulfur-containing heterocyclic compounds such as (2R, 4S) - (-) - N- [4- (diethoxyphosphorylmethyl) phenyl] -1,2,4,5-tetrahydro-4-methyl -7,8-methylenedioxy-5-oxo-3-benzothiepin-2-carboxamide or its salts described in US5071841, US5158943 and JP529460, benzopyran derivatives such as N- (4-dimethoxyphosphorylmethylphenyl) -4-oxo-4H-1 -benzopyran-2-carboxamide or its salts described in EP625522, phosphonic acid derivatives such as diethyl-4- (7-cyclohexyl-3,4-dihydro-2-naphthalene boxamide) benzyl phosphonate or its salts described in W096 / 01267, prostaglandin derivatives A ₁ described in Journal of Pharmacology and Experimental Therapeutics, vol. 258, pp. 1120-1126 (1991), vitamin D ₃ derivatives described in Bioorganic and Medicinal Chemistry Letters, vol. 3, pp. 1815-1819 (1993), benzylphosphonic acid derivatives described in EP524023, bisphosphonic acid derivatives described in Bone, vol.13, pp.249-255 (1992), and vitamin K ₂ derivatives described in Biochemical and Biophysical Research Communicatios, vol. 187, pp. 814-820 (1992).

 The pharmaceutical composition of the present invention may contain one or more non-peptide osteogenic activating agents described above as an active ingredient.

 In the aforementioned non-peptide osteogenesis activating optically active substances, a compound represented by the formula (I) or a salt thereof is preferably used for the present invention.

где R₃ представляет низшую алкильную группу; R₄ и R₅ независимо представляют низшую алкильную группу или связываются вместе с образованием низшей алкиленовой группы.An optically active compound represented by formula (I):

where R ₃ represents a lower alkyl group; R ₄ and R ₅ independently represent a lower alkyl group or bind together to form a lower alkylene group.

может обозначать группу:

где р равно целому числу от 2 до 4.Examples of the lower alkyl group represented by R ₃ , R ₄ or R ₅ in the formula (I) are alkyl groups having 1-6 (preferably 1-4) carbon atoms, such as methyl, ethyl, propyl, isopropyl, butyl, isobutyl , sec-butyl, tert-butyl, pentyl, isopentyl, neopentyl and hexyl. R ₄ and R ₅ can bind together to form a lower alkylene group. In this case, part of the molecule:

may indicate a group:

where p is an integer from 2 to 4.

Preferred groups for R ₃ , R ₄ and R ₅ include alkyl groups having 1-4 carbon atoms, such as methyl and ethyl. Compound (I) is an optically active compound (2R, 4S) of the configuration and essentially does not contain the compound (2S, 4R) of the configuration. Compound (I) with an optical purity close to 100% is preferred.

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

 The salt of the non-peptide osteogenic activating compound of this invention is preferably a pharmaceutically acceptable salt. 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 hydroxides of alkali metals (eg, sodium, potassium) and alkaline earth metals (eg, calcium, magnesium), such organic bases include trimethylamine, triethylamine, pyridine, picoline, N, N-dibenzylethylenediamine and diethanolamine such inorganic acids include hydrochloric acid, hydrobromic acid, hydroiodic acid, phosphoric acid, nitric acid and sulfuric acid, such organic acids include formic acid, acetic acid, trifluoroacetic 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.

где R₃ имеет значения, приведенные выше, или реакционноспособного производного по карбоксильной группе или его соли с соединением формулы (III):

где R₄ и R₅ имеют значения, приведенные выше, его реакционноспособным производным по аминогруппе или его солью.The non-peptide osteogenic activating optically active substance of the present invention can be obtained, for example, by a well-known method (for example, according to US5071841, US5158943 described above) and by the method described below, or its modification. For example, compound (I) or a salt thereof can be prepared by reacting an optically active compound of formula (II):

where R ₃ has the meanings given above, or a reactive derivative of a carboxyl group or its salt with a compound of formula (III):

where R ₄ and R ₅ have the meanings given above, its reactive derivative at the amino group or its salt.

 Preferred reactive derivatives of the amino group on compound (III) include tautomeric isomers of the imino or enamino form of Schiff bases resulting from the reaction of compound (III) and a carbonyl compound such as an aldehyde (eg, acetaldehyde) or ketone (eg, acetone); silyl derivatives resulting from the interaction of compound (III) and a silyl compound such as bis (trimethylsilyl) acetamide, mono (trimethylsilyl) acetamide or bis (trimethylsilyl) urea; and derivatives resulting from the interaction of compound (III) and phosphorus trichloride or phosgene.

 Preferred reactive derivatives of the carboxy group on compound (II) include acid halides, acid anhydrides, activated amides and activated esters, which are prepared according to conventional methods. Preferred reactive derivatives include acid chlorides; acid azides; mixed acid anhydrides with substituted phosphoric acid, such as dialkylphosphoric acid, phenylphosphoric acid, diphenylphosphoric acid, dibenzylphosphoric acid or halogenated phosphoric acid, dialkylphosphoric acid, sulfuric acid, thiosulfuric acid, sulfuric acid, sulfonic acid, sulfonic acid, sulfonic acid such as acetic acid, propionic acid, butyric acid, isobutyric acid, pivalic acid, pentanoic acid, isopentanoic acid one or trichloroacetic acid or aromatic carboxylic acid such as benzoic acid; symmetric acid anhydrides; activated amides with imidazole, 4-substituted imidazole, dimethylpyrazole, triazole or tetrazole; activated esters such as cyanomethyl ether, methoxymethyl ether, dimethyliminomethyl ether, vinyl ether, propargyl ether, p-nitrophenyl ether, trichlorophenyl ether, pentachlorophenyl ether, mesylphenyl ether, phenylthiophenyl ether, phenyl ether, phenyl ether, ether pyranyl ether, pyridyl ether, piperidyl ether or 8-quinolylthioether; and esters of N-hydroxy compounds such as N, N-dimethylhydroxylamine, 1-hydroxy-2- (1H) -pyridone, N-hydroxysuccinimide, N-hydroxyphthalimide, 1-hydroxy-1H-benzo-triazole and N-hydroxy-5 norbornene-2,3-dicarboxyimide. These reactive derivatives may optionally be selected according to the type of compound (II).

 Preferred salts of the reactive derivatives of compound (II) or (III) include basic salts, for example alkali metal salts such as sodium salt and potassium salt, alkaline earth metal salts such as calcium and magnesium salt, ammonium salt, and organic base salts, such as a trimethylamine salt, triethylamine salt, pyridine salt, picoline salt, dicyclohexylamine salt and dibenzylethylenediamine salt.

This reaction is usually carried out in a conventional solvent, such as water, alcohol (e.g. methanol, ethanol), acetone, dioxane, acetonitrile, chloroform, methylene chloride, ethylene chloride, tetrahydrofuran, ethyl acetate, N, N-dimethylformamide or pyridine, but it can be carried out in any other organic solvent, if it does not interfere with the reaction. These common solvents may be used in admixture with water. In this reaction, when using compound (II) or (III) in the form of a free acid or its salt, the reaction is preferably carried out in the presence of a conventional condensing agent, such as 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); pentamethylene ketene-N-cyclohexylimine; diphenylketene-N-cyclohexylimine; ethoxyacetylene; 1-alkoxy-1-chloroethylene; trialkylphosphite; ethyl polyphosphate; isopropyl polyphosphate; phosphorus oxychloride; diphenylphosphoryl azide; thionyl chloride; oxalyl chloride; lower alkyl haloformate such as ethyl chloroformate or isopropyl chloroformate; triphenylphosphine; 2-ethyl-7-hydroxybenzisoxoxazolium salt; intramolecular salt of 2-ethyl-5- (m-sulfophenyl) isoxazolium hydroxide; N-hydroxy-benzotriazole; 1- (p-chlorobenzenesulfonyloxy) -6-chloro-1H-benzotriazole or a Wilsmeier reagent obtained by reacting N, N'-dimethylformamide with thionyl chloride, trichloromethyl chloroformate or phosphorus oxychloride. A method using a condensing agent such as N, N'-dicyclohexylcarbodiimide in the presence of N-hydroxybenzotriazole or N-hydroxy-5-norbornene-endo-2,3-dicarboxyimide is also desirable. This reaction can also be carried out in the presence of an inorganic or organic base such as alkali metal bicarbonate, tri (lower) alkylamine, pyridine, N- (lower) alkylmorpholine or N, N-di (lower) alkylbenzylamine. Although the reaction temperature is not limited, this reaction is usually carried out under cooling - heating (-10 -120 ^o C). The reaction time is usually about 0.5-100 hours, preferably about 1-50 hours.

 Compound (I) thus obtained can be isolated and purified by known separation and purification methods, such as concentration, concentration under reduced pressure, solvent extraction, crystallization, recrystallization, re-dissolution and chromatography.

 The starting compound (II) can be obtained, for example, by optical resolution of the racemate of compound (II) described in US5158943. In particular, an optically active compound is prepared by preparing a racemate salt of compound (II) and an optically active base (e.g., α-methylbenzylamine, brucin, quinine, cichonin), repeating fractional crystallization based on the difference in solubility between the resulting diastereoisomers to obtain a slightly soluble salt in its pure form followed by acid treatment. Another optically active compound can be obtained by esterification of the racemate of compound (II) with an optically active alcohol (e.g., optically active methyl lactate, methyl mandelate), obtaining the other ester in pure form based on the difference in physical properties between the resulting diastereoisomers, followed by hydrolysis.

 The copolymer of lactic acid and glycolic acid according to this invention is a copolymer that is soluble or insoluble in water and is degradable in vivo.

 The polymerization may be random polymerization, block copolymerization or graft polymerization.

 The average molecular weight of these copolymers of this invention is preferably selected from the range of from about 2000 to 800000, more preferably from about 5000 to 200000.

 The relative content of lactic acid and glycolic acid in the lactic acid-glycolic acid copolymer is preferably about 100 / 0-50 / 50 by weight, and more preferably about 90 / 10-50 / 50 by weight. The weight average molecular weight of the lactic acid-glycolic acid copolymer is preferably about 5000 to 100000, more preferably about 8000-50000. The lactic acid-glycolic acid copolymer can be synthesized by a well-known production procedure, such as described in EP172636. This copolymer is preferably synthesized by dehydrogenation polycondensation in the absence of a catalyst.

 In this application, weight average molecular weight is defined as weight average weight based on polystyrene, measured by gel permeation chromatography (GPC). The measurements were carried out using a KF804L • 2 GPC column (manufactured by Showa Denko) and an RI L-3300 monitor (manufactured by Hitachi Ltd.) with chloroform as the mobile phase.

 The amount of this copolymer varies in accordance with the pharmacological activity of the non-peptide osteogenesis of the activating substance, the speed and duration of the release of the drug from the copolymer, and so on, until the desired goal is achieved. For example, the copolymer is used in about 0.2-10000-fold amounts (weight ratio), preferably about 1-1000-fold amounts, more preferably about 1-100-fold amounts, relative to the amount of bioactive substance.

 The pharmaceutical composition of the present invention can be obtained by conventional methods for preparing the pharmaceutical composition, for example, it can be obtained by dispersing a non-peptide activating substance in a given copolymer or placing a non-peptide osteogenic activating substance in a preformed hollow copolymer. In particular, the methods used include a method of drying in water, a method of phase separation, a method of spray drying and their modification.

 The pharmaceutical composition of the present invention obtained by the method described above may be in the form of, for example, small particles, spheres, sticks, needles, granules, films or creams, but its form is not limited only to these forms to achieve the desired goal.

 In this application, the pharmaceutical composition of fine particles is also called microcapsules or microspheres.

 Examples of methods for producing microcapsules are described below.

 (1) A method of drying in water (m / v, i.e., a method for producing an oil-in-water emulsion).

In this method, a solution in an organic solvent containing a copolymer is first prepared. The organic solvent used to prepare the pharmaceutical composition of this 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 ether, butyl ether), ethers (e.g. ethyl ether, isopropyl ether) and aromatic hydrocarbons (e.g. benzene, toluene, xylene). These solvents can be used in combination of two or more types of solvents in suitable ratios. A preferred solvent is dichloromethane or acetonitrile, more preferably dichloromethane. The concentration of the copolymer in an organic solvent solution is usually selected in the range of from about 0.01 to 80 wt. %, preferably about 0.1-70 wt.% and more preferably about 1-60 wt.%, although it varies depending on the molecular weight of the copolymer and the type of solvent, etc.

 The non-peptide osteogenic activating substance is added and dissolved in the solution thus obtained in an organic solvent containing the copolymer, if necessary after lyophilization or drying in vacuo. The amount of non-peptide osteogenic activating agent is from about 0.001 to 90% by weight, preferably from about 0.1-80% by weight, and more preferably from about 0.1-50% by weight, based on the concentration of the copolymer in the organic solvent solution, although the amount varies depending on the type of drug, its mechanism of action on osteogenesis, duration of action, etc.

 The solution thus obtained in an organic solvent is then added to the aqueous phase to obtain an oil-in-water emulsion using a turbine drive mixer or the like. The volume of the aqueous phase is usually selected from a range of from about 1 to 10,000 times, preferably from about 2 to 5,000 times, and more preferably from about 5 to 2,000 times relative to the volume of the oil phase.

 An emulsifier may be added to the aqueous phase. It can be any emulsifier if it is able to form a stable oil-in-water emulsion. Examples of such emulsifiers include anionic surfactants, nonionic surfactants, castor oil polyoxyethylene derivatives, polyvinylpyrrolidone, polyvinyl alcohol, carboxymethyl cellulose, lecithin, gelatin and hyaluronic acid. They can be used in a suitable combination. The concentration of emulsifier in the aqueous phase is preferably approximately 0.001-20 wt.%, More preferably approximately 0.01-10 wt. % and even more preferably approximately 0.05-5 wt.%.

 Evaporation of the solvent from the oil phase can be carried out by generally accepted methods, including by the method in which the solvent is evaporated under normal or gradually decreasing pressure while stirring with a propeller stirrer or magnetic stirrer, etc., and by the method in which the solvent is evaporated at bringing the vacuum to the desired value using a rotary evaporator, etc. The obtained microcapsules are separated by centrifugation or filtration, after which they are washed, for example, with water or heptane, several times to remove free non-peptide osteogenic activating substances, emulsifier, etc., adhered to the surface of the microcapsules. Then the microcapsules are again dispersed in distilled water, etc. and lyophilized.

 In the above m / v method, microcapsules can be obtained by the r / m / b method (solvent / oil / water), in which the non-peptide osteogenic activating substance is dispersed in an organic solvent solution containing this copolymer.

(2) Method of drying in water (v / m / v method)
In this method, the non-peptide osteogenic activating substance or its salt is first dissolved or dispersed in water to obtain the above concentration to obtain the internal aqueous phase, if necessary, with dissolution or suspension, adding a drug-retaining substance such as protein (e.g. gelatin), seaweed (e.g. agar), polysaccharide (e.g. alginic acid), synthetic high molecular weight substance (e.g. polyvinyl alcohol), basic amino acid (e.g. arginine, Isigny) 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 their salt (for example, 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 to maintain stability and solubility of the non-peptide osteogenic activating optically active substance or its salt. As a stabilizer of a non-peptide osteogenic activating optically active substance, a protein (e.g., albumin, gelatin), a starch derivative (e.g., dextrin, pullulan, etc.), an organic acid (e.g., citric acid), an alkali metal salt of ethylenediaminetetraacetic acid can be added. (e.g. sodium ethylene diamine tetraacetate), an acidic alkali metal salt of sulphurous acid (e.g. sodium bisulfite), a synthetic high molecular weight substance (e.g., polyethylene glycol) or the like. There may also be Bavleny conventional preservatives such as p-hydroxybenzoates (e.g., methylparaben, propylparaben), benzyl alcohol, chlorobutanol and thimerosal. An additional amount of non-peptide osteogenic activating substance is approximately 0.001-90 wt.%, Preferably approximately 0.01-80 wt. %, more preferably about 0.1-50 wt. %, although this amount varies depending on the type of drug, its mechanism of action on osteogenesis or duration of action, etc.

The resulting internal aqueous phase is added to the solution (oil phase) containing this copolymer, followed by emulsification treatment to obtain an emulsion of the type water-in-oil. Emulsification is achieved by known dispersion methods, which include a discontinuous shaking method, a method using a mixer, such as a propeller mixer or a turbine mixer, a method using a colloid mill, a method using a homogenizer, and an ultrasonic treatment method. The above-described solution (oil phase) containing this copolymer is a solution obtained by dissolving the copolymer in an organic solvent. This solvent may be any solvent if its boiling point does not exceed about 120 ^{° C.} and if it does not mix with water. Such solvents include halogenated hydrocarbons (e.g., dichloromethane, chloroform, chloroethane, dichloroethane, trichloroethane, carbon tetrachloride), aliphatic esters (e.g. ethyl acetate, butyl acetate), ethers (e.g. ethyl ether, isopropyl ether) and aromatic hydrocarbons ( e.g. benzene, toluene, xylene). These solvents can be used in combination of two or more types in suitable proportions.

 Then, the resulting water-in-oil emulsion is added to the aqueous phase to obtain a water / oil / water emulsion from which the solvent of the oil phase is evaporated to give microcapsules. The specific method for this preparation is identical to the method described in (1) above.

(3) Phase Separation Method
In this method, a coacervating agent is gradually added to the above-described water / oil emulsion with stirring to precipitate and solidify the copolymer. Silicone oils, vegetable oils and fats (e.g., sesame oil, soybean oil, corn oil, cottonseed oil, linseed oil), mineral oils, hydrocarbons (e.g. n-hexane, n-heptane) can be used as a coacervating agent, if they are polymer compounds of mineral oil or vegetable oil that can be mixed with a solvent of the copolymer and which do not dissolve the copolymer to encapsulate. They can be used in combination of two or more types of coacervating agents.

 The resulting microcapsules, after filtering and separating, are repeatedly washed with heptane, etc., to remove the coacervating agent. Then the free drug and solvent are removed using the same drying method in water. Antiflocculants can be added during washing to prevent flocculation of particles: 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 .

(4) Spray Drying Method
To obtain microcapsules in this method, the above-described water-in-oil emulsion is sprayed through a spray nozzle into a spray drying oven to evaporate the organic solvent and water in the form of small droplets in a short time and microcapsules are obtained. Examples of a spray nozzle are a two-component nozzle, a high pressure nozzle, and a rotary disk nozzle. To prevent flocculation of the microcapsules through another nozzle, an aqueous solution of the anticoagulant described above can be sprayed simultaneously with the atomization of a water-in-oil emulsion. The microcapsules thus obtained can be heated under reduced pressure to facilitate the removal of the water and solvent contained therein.

 In addition to obtaining the above-described microcapsules, a pharmaceutical composition according to this invention can be obtained by dissolving a non-peptidic osteogenic activating optically active substance dispersed in this copolymer and forming this solution into spheres, needles, granules, films or the like. using the appropriate method for this. A non-peptide osteogenic activating optically active substance dispersed in a copolymer is obtained, for example, according to the method described in US3773919.

 In addition, the pharmaceutical composition of the present invention can also be obtained by spraying to a suitable particle size a non-peptide osteogenic activating optically active substance dispersed in the copolymer according to a method such as described in JP62234656 using a turbine-type jet mill sprayer or an ultrasonic jet sprayer. In particular, a non-peptide osteogenic activating optically active substance is added to and dissolved in an organic solvent containing a copolymer. Then, the solid solution obtained by vacuum drying is coarsely ground and sieved, followed by removal of the solvent, after which coarse particles are ground to a controlled particle size using an ultrasonic jet spray to obtain a pharmaceutical composition of the present invention.

 The pharmaceutical composition of the present invention can be used in the form of microcapsules as such, or formulated in various dosage forms with microcapsules, spheres, sticks, needles, balls, films or creams as starting material. The pharmaceutical composition of the present invention may contain phosphoric acid or a salt thereof (e.g., sodium phosphate, potassium phosphate) in an amount of about 0-30%. The pharmaceutical composition of the present invention may also be administered as a parenteral agent for topical administration (for example, as injectable preparations for intramuscular, subcutaneous, administration to organs or joints, etc., solid preparations, such as permanently administered preparations, granules and powders liquid preparations such as suspensions and ointments).

 The injectable preparation can be prepared as an aqueous suspension by suspending the microcapsules in water together with a dispersing agent (for example, surfactants such as Tween 80 and HCO-60, polysycharides such as carboxymethyl cellulose, sodium alginate and hyaluronic acid and polysorbate), preservative (e.g. methylparaben, propylparaben), an isotonic agent (e.g. sodium chloride, mannitol, sorbitol, glucose), a buffer (e.g. calcium carbonate), a pH adjusting agent (e.g. sodium phosphate, potassium phosphate) etc., and it can be prepared as an oily suspension by dispersing the microcapsules in a vegetable oil, such as sesame oil or corn oil, with or without phospholipid such as lecithin or a moderate chain fatty acid triglyceride (e.g. MIGLYOL 812).

 Phosphate can increase the osteogenic activating activity of the pharmaceutical composition according to this invention.

 The concentration of sodium phosphate or potassium phosphate in such an injectable preparation is about 0.1 mM to 500 mM, preferably about 1 mM-100 mM.

 A preferred embodiment of the invention is the following.

(A) a lactic acid-glycolic acid copolymer in which the ratio of lactic acid / glycolic acid is about 90 / 10-50 / 50 by weight and the weight average molecular weight is about 8000-50000,
(B) (2R, 4S) - (-) - N- [4- (diethoxyphosphorylmethyl) phenyl] -1,2,4,5-tetrahydro-4-methyl-7,8-methylenedioxy-5-oxo-3- benzothiepin-2-carboxamide and (C) sodium phosphate.

 The relative content (B) based on (A) is approximately 30% by weight.

 The relative content of (C) based on (A) and (B) is approximately 0.1-20 wt.%.

 When using microcapsules, for example, as an injectable suspension, their particle size is selected in the range of about 0.1-300 μm (average particle diameter) if they satisfy the requirements regarding the degree of dispersion and the ability to pass through the syringe needle. Preferably, the particle size is about 1-150 microns, more preferably about 2-100 microns.

 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 due to the fact that such a composition will cause less pain in a patient when administered through an injection needle for subcutaneous or intramuscular injection.

 The pharmaceutical composition of the present invention is preferably an injectable preparation.

 Methods for producing microcapsules in the form of a sterile preparation include, but are not limited to, a method in which the entire preparation process is sterile, a method in which gamma radiation is used as the sterilizing agent, and a method in which an antiseptic agent is added.

The pharmaceutical composition of this invention can be used to prevent and treat bone diseases (e.g., bone fractures, refractures, osteoporosis, osteomalacia, Behcet's syndrome (triple symptom complex), ankylosing spondylitis, rheumatoid arthritis and destruction of joint tissue caused by deforming gonarthritis, and genus) , to restore bone tissue after surgery for myeloma (Caler's disease), lung cancer, breast cancer, etc. and for the regeneration of periodontal tissue in periodontopathy, since this composition exhibits a property of sustained (delayed) release with increased activity of a non-peptide osteogenic activating substance and has a time of prolonged release from 1 week to 3 months depending on the type and content of the copolymer, etc. The pharmaceutical composition of the present invention is particularly effective in the case of patients with bone fractures, as these patients usually have a fixed affected part covered with a plaster cast and wish to more readily accelerate recovery through a single administration than through multiple administrations. A sustained release preparation consisting of the pharmaceutical composition of the present invention can be used in combination with other active agents. For example, in the case of the compound represented by formula (I), calcium preparations (e.g., calcium carbonate), calcitonin, vitamin D (e.g., alfacalcidol), sex hormone (e.g., estrogen, estradiol, can be used as an osteogenic activating agent for combination with it) ), prostaglandin A ₁ , bisphosphonic acid, ipriflavone, a fluorine compound (e.g. sodium fluoride), vitamin K ₂ , BMP (bone morphogenetic protein), FGF (fibroblast growth factor), PDGF (platelet growth factor), TGF-β (transforming growth factor-β), IGF-1 (ins Uline-like growth factor-1), IGF-2 (insulin-like growth factor-2), PTH (parathyroid hormone) and so on.

 With low toxicity, the pharmaceutical composition of this invention can be safely used for mammals (for example, for humans, cattle, horses, pigs, cats, mice, rats, rabbits).

 It is expected that the pharmaceutical composition of the present invention can serve as a safe drug with high efficacy, providing a constant effect of the drug with low toxicity and meeting the requirements of prevention and treatment of bone diseases, restoration of damaged bone tissue and regeneration of periodontal tissue in case of periodontitis, etc. because this composition constantly releases the drug over a long period of time. For example, when using the pharmaceutical composition of the present invention for the treatment of bone fractures (e.g., femoral neck fractures), it can effectively exhibit its osteogenic activating effect locally and significantly reduce the recovery time, which is usually 2-6 months after a bone fracture. Therefore, patients quickly return to normal social life and various complications caused by bone fractures in old age can also be prevented.

 The dose of the pharmaceutical composition of this invention may be an effective amount of a non-peptide osteogenic activating optically active substance, however, depending on the type and content of this non-peptide osteogenic activating optically active substance, the release time of the drug, a particular animal, etc. For example, when using the pharmaceutical composition of the present invention in the form of microcapsules for treating a bone site with a fracture can be administered at a dose of approximately flax from 0.01 to 500 mg, preferably about 5-50 mg, based on the content of the active ingredient (e.g., compound (I)) per adult patient (weighing 50 kg) per dose, once a week to once per month.

Brief Description of Drawings
FIG. 1 is a graphical depiction of the mineral content (mg) of the fibula 2 weeks after the administration of Compound A in the form of microcapsules and placebo microcapsules (control) to rats.

 FIG. 2 is a graphical representation of a temporary change in the retention rate of compound A at the injection site in rats receiving subcutaneous implantation of a tablet containing compound A in the form of microcapsules in the back. The abscissa indicates the time (weeks) after administration. The ordinate shows the remaining dose (%) of compound A.

 FIG. 3 is a graphical representation of the temporal change in the concentration of compound A in the blood of rats that received subcutaneous implantation in the backs of compound A microcapsules.

 The abscissa indicates the time (weeks) after administration. The ordinate shows the concentration (μg / ml) of compound A in the blood.

BEST MODE FOR CARRYING OUT THE INVENTION
The invention is further described in more detail using the following cited as reference examples, working examples and test examples, which are not intended to limit the invention. In working examples and test examples, room temperature refers to a temperature between approximately 0 and 30 ^o C.

Examples
Reference example 1
Preparation of (R) -α-methoxycarbonylbenzyl ether (2R, 4S) - (-) - 1,2,4,5-tetrahydro-4-methyl-7,8-methylenedioxy-5-oxo-3-benzothiepin-2-carboxylic acid
A solution of 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide hydrochloride (12.59 g) in dichloromethane (200 ml) was added dropwise to a solution of (±) -t-1,2,4,5-tetrahydro-4-methyl -7,8-methylenedioxy-5-oxo-3-benzothiepin-2-carboxylic acid (15.34 g) and methyl- (R) - (-) - mandelic acid (18.19 g) in N, N-dimethylformamide (DMF) (200 ml) at 0 ^{° C.} followed by the addition of 4-dimethylaminopyridine (DMAP) (3.34 g). After stirring at 0 ^{° C.} for 1 hour and at room temperature for 15 hours, the mixture was poured onto water and extracted with ethyl acetate. The ethyl acetate layer was washed with water and dried (MgSO ₄ ), after which the solvent was evaporated. The remaining crystals were collected by filtration, washed with ether-hexane and recrystallized twice from ethyl acetate-hexane to give the title compound (4.09 g, 17% yield).

Melting point: 140-141 ^o C.

Optical rotation [α] _D (23 ^° C): -244.2 ^° C (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-benzothiepin-2-carboxylic acid
Mixture of (R) -α-methoxycarbonylbenzyl ether (2R, 4S) - (-) - 1,2,4,5-tetrahydro-4-methyl-7,8-methylenedioxy-5-oxo-3-benzothiepin-2-carboxylic the acid obtained in Reference Example 1 (4.18 g), acetic acid (45 ml) and concentrated hydrochloric acid (30 ml) were stirred for 30 minutes under reflux. The reaction mixture was poured into water (800 ml). The resulting crystalline material was collected by filtration and dissolved in ethyl acetate (150 ml). The ethyl acetate layer was washed with water and dried (MgS0 ₄ ), after which the solvent was evaporated. The remaining crystals were collected by filtration, washed with hexane and recrystallized from ethyl acetate-hexane to give the title compound (1.62 g, 59% yield) in the form of colorless needle crystals.

Melting point: 194-195 ^o C.

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

Раствор гидрохлорида 1-этил-3-(3-диметиламинопропил)карбодиимида (0,39 г) в дихлорметане (7 мл) добавляли по каплям к раствору (2R,4S)-(-)-1,2,4,5-тетрагидро-4-метил-7,8-метилендиокси-5-оксо-3-бензотиепин-2-карбоновой кислоты (0,47 г), полученной в справочном примере 2, и диэтил 4-аминобензилфосфоната (0,41 г) в N,N-диметилформамиде (ДМФ) (7 мл) при 0^oС с последующим добавлением 1-гидроксибензотриазола (HOBt) (0,28 г). После перемешивания при 0^oС в течение 1 часа и при комнатной температуре в течение 15 часов смесь выливали на воду и экстрагировали этилацетатом. Слой этилацетата промывали водой и сушили (MgSO₄), после чего растворитель выпаривали. Оставшиеся кристаллы собирали фильтрованием и перекристаллизовывали из смеси этилацетат-гексан и смеси метанол-гексан с получением соединения А (0,37 г, выход 44%) в форме бесцветных призм.Reference example 3
Preparation of (2R, 4S) - (-) - N- [4- (diethoxyphosphorylmethyl) phenyl] -1,2,4,5-tetrahydro-4-methyl-7,8-methylenedioxy-5-oxo-3-benzothiepin 2-carboxamide (compound A)

A solution of 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide hydrochloride (0.39 g) in dichloromethane (7 ml) was added dropwise to a solution of (2R, 4S) - (-) - 1,2,4,5-tetrahydro -4-methyl-7,8-methylenedioxy-5-oxo-3-benzothiepin-2-carboxylic acid (0.47 g) obtained in Reference Example 2, and diethyl 4-aminobenzylphosphonate (0.41 g) in N, N-dimethylformamide (DMF) (7 ml) at 0 ^{° C.} followed by the addition of 1-hydroxybenzotriazole (HOBt) (0.28 g). After stirring at 0 ^{° C.} for 1 hour and at room temperature for 15 hours, the mixture was poured onto water and extracted with ethyl acetate. The ethyl acetate layer was washed with water and dried (MgSO ₄ ), after which the solvent was evaporated. The remaining crystals were collected by filtration and recrystallized from ethyl acetate-hexane and methanol-hexane to give compound A (0.37 g, 44% yield) in the form of colorless prisms.

Melting point: 181-182 ^o C.

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

Example 1
A dichloromethane solution was prepared of a lactic acid-glycolic acid copolymer, hereinafter referred to as PMHC; the relative content of lactic acid-glycolic acid (mol.%) and weight average molecular weight according to GPC are shown in table 1; manufactured by Wako Pure Chemical Industry. Using the formula shown in table 1 (the solution is hereinafter referred to as solution A). Similarly, a solution in dichloromethane of compound A was also prepared using the formula shown in table 1 (the solution is hereinafter referred to as solution B). Solutions A and B were uniformly mixed
together. This mixture was introduced into a 0.1% aqueous solution of polyvinyl alcohol (EG-40 manufactured by Nippon Synthetic Chemical Industry) (hereinafter referred to as PVA solution), cooled to 15 ^{° C.} in advance, in the volume indicated in table 1, and emulsified using a homo-mixer turbine type at 7000 rpm to obtain an oil-in-water emulsion, which was then stirred at room temperature for 3 hours to evaporate dichloromethane. After curing, the oil phase was centrifuged at 2000 rpm using a centrifuge (05PR-22, Hitachi Ltd.). The collected microcapsule fraction was again dispersed in distilled water, then centrifuged, followed by washing of the released drug, etc. The collected microcapsules were again dispersed in a small amount of distilled water and then lyophilized. Received microcapsules with particle sizes shown in table 1. The content of microcapsules, including compound A, was 100%.

Example 2
500 mg of the microcapsules 1 obtained in Example 1 were uniformly dispersed in two test tubes of a Tisseel fibrinogen solution (manufactured by Nippon Zoki Pharmaceutical). A thrombin solution for Tisseel was gradually added to the two test tubes. Then each mixture was immediately filled into a plastic syringe. The syringe was allowed to stand at 37 ^{° C.} for 30 minutes to cure the contents. After curing, the contents were squeezed out of the tip of the syringe and cut with a razor into granules with a volume of approximately 200 μl.

Example 3
To the microcapsules 3 obtained in Example 1, which contain compound A (relative content 4%), 20% crushed gelatin (manufactured by Nitta Gelatin) was added to obtain a microcapsule-containing preparation in the form of a 5.5 mm diameter tablet and weighing 125 mg.

Example 4
Microcapsules 7 containing compound A (relative content 10%) were obtained in the same manner as in Example 1, except that PLGA (PMGA) had a relative content of lactic acid-glycolic acid 85/15 (mol%) and weight average molecular weight weight 14900 (Wako Pure Chemical Industry). The average particle size was 31 microns.

Example 5
A solution in dichloromethane containing 2.4 g of PLGA (PMHC) (Wako Pure Chemical Industry), the relative content of lactic acid / glycolic acid of which is 85/15 and the weight average molecular weight is 14900, and 0.1 g of compound A, were also obtained as described in Example 1. 0.2 g of estradiol was added to the solution. Then a PVA solution was added to this solution to obtain an oil-in-water emulsion. Microcapsules 8 containing compound A and estradiol were obtained.

 The average particle size was 27 microns.

Test Example 1
500 mg of the microcapsules 1 obtained in Example 1 were accurately weighed and transferred to a glass centrifuge tube and allowed to stand at 37 ^{° C.} in powder form. After a certain time, the microcapsules were dissolved in a small amount of acetonitrile and quantitatively analyzed by HPLC. Stability tests are presented in table 2.

 The drug content was more than 95% even after 4 weeks.

Test Example 2
The microcapsules 2 and 3 obtained in Example 1, 5 mg each, were precisely weighed and transferred to a glass vessel, followed by shaking at 37 ^{° C.} in a water bath (TAITEC, thermostat M-100, 115 bpm) in the presence of 10 ml test release solution (phosphate buffer supplemented with 10% bovine serum albumin, pH 7.0). To each 100 μl sample taken after a certain time, 100 μl of acetonitrile was added. After shaking, the mixture was centrifuged. The resulting supernatant was analyzed by HPLC to determine the amount of released compound A. The release tests are presented in table 3.

 Three types of different release distributions were obtained.

Test Example 3
Microcapsules 3-5 obtained in example 1, 25 mg each, were dispersed in 0.3 ml of a dispersing solution (solution of 1.5 mg of carboxymethyl cellulose, 0.3 mg of polysorbate 20 and 15 mg of mannitol in distilled water) and injected subcutaneously into the heads male SD rats (n = 5) at 5 weeks of age under ether anesthesia with a 22G injection needle. Rats were euthanized at predetermined time intervals after administration. The microcapsules remaining at the injection site were removed and analyzed by HPLC to determine the amount of compound A in the microcapsules. The results are presented in table 4.

 Also in vivo, various types of microcapsule release distribution have been obtained. In particular, it was found that microcapsules 3 have a sustained release for 1 month, microcapsules 4 for more than 1 month, and microcapsules 5 for 3 weeks.

Test Example 4
During anesthesia with pentobarbital, the heads of male SD rats aged 6 weeks were incised and the periosteum (periosteum) was separated. Then a hole was made with a diameter of 4 mm in the left cranial vault using a dental drill. 1 week after suturing, 25 mg of the microcapsules 3 obtained in Example 1 were dispersed in 0.3 ml of a dispersing agent (a solution of 1.5 mg of carboxymethyl cellulose, 0.3 mg of polysorbate 20 and 15 mg of mannitol in distilled water) and injected subcutaneously into the right temple with a 22G injection needle. For control, rats were used who received 0.3 ml of the above dispersant solution. Three weeks later, rats were euthanized. The cranial vault was cut and subjected to soft x-ray analysis. Using the obtained photograph, the area of newly formed bone tissue in the damaged area of the bone was determined by image analysis.

 A significant increase in the area of newly formed bone tissue was noted in the group with the introduction of microcapsules, which indicates excellent osteogenesis of the activating ability of the pharmaceutical composition according to this invention.

Test Example 5
During anesthesia with pentobarbital, the left limbs of male SD rats (n = 8) at the age of 6 weeks were incised and the central part of the left fibula was exposed and then cut with a cutting tool. 25 mg of the microcapsules 1 obtained in Example 1 were immersed in a section of the fibula incision followed by suturing. Two weeks later, rats were killed. The fibula was excised and analyzed using a bone mineral content analyzer (DSC-600, Aloca Co., Ltd., Tokyo) to determine bone mineral content. The bone mineral content of the untreated right fibula was also determined. The measurement for the right fibula was subtracted from the measurement for the left fibula to obtain the contents of the mineral components in the callus. To control the microcapsules, placebo without compound A was prepared according to the method used to obtain microcapsules 1, and compared with capsules containing A in the same way as in Test Example 4. These results are shown in FIG. 1.

 The administration of drug-containing microcapsules resulted in significantly increased bone mineral content, demonstrating the excellent osteogenic activating ability of the pharmaceutical composition of this invention.

Test Example 6
25 mg of microcapsules 1 containing compound A (relative content 4%) obtained in Example 1 were injected locally into rat models with a fracture of the fibula according to the method described in Test Example 5. After 2 and 3 weeks, the bone mineral content was determined. For control, the bone mineral content was determined in the group receiving microcapsules with placebo (drug-free) and in the spontaneous healing group. These results are shown in table 5.

 It was found that the group receiving the microcapsules containing the drug had a significantly higher mineral content compared with the control groups, which indicated the excellent osteogenic activating ability of the pharmaceutical composition according to this invention.

Test Example 7
The microcapsule-containing tablet preparation obtained in Example 3 was implanted subcutaneously in the backs of male SD rats (n = 8) at 5 weeks of age under ether anesthesia. The amount of drug remaining at the injection site was determined by HPLC over time. The results are presented in figure 2. The test tablet found a sustained release property for 6 weeks.

Test Example 8
In accordance with the method of Miyamoto et al. "Shinpei Miyamoto, Hideki Yoshikava and Kunio Takaoka: The Bone, 7, 85-96 (1983)" the tibia of rabbits (3-4 kg in weight) was cut to form a 5 mm bone lesion, which was then filled with a microcapsule-containing preparation in the form tablets obtained in example 3, and then the wound was fixed outside. For control, a tablet preparation was used containing microcapsules with a placebo that did not contain Compound A. A soft X-ray radiographic analysis revealed bone healing in rabbits who were given a pill containing Compound A 2 months after surgery, whereas in rabbits treated with a pill with a placebo, healing was not seen.

Test Example 9
Microcapsules 7 containing compound A (relative content of 10%) obtained in Example 4 were injected subcutaneously into the backs of male SD rats (n = 5) at 5 weeks of age under ether anesthesia according to the method described in Test Example 2 (100 mg / kg rat body weight calculated on compound A). The concentration of the drug in the blood was determined using HPLC over time. The results are shown in FIG. Even at 4 weeks after administration, a drug concentration in the blood of 0.05-0.1 μg / ml was obtained.

Test Example 10
A rat model of fibula fracture was obtained according to the method described in Test Example 5. This fracture model was administered topically with microcapsules 7 containing compound A (10% relative content) obtained in Example 4 in the form of lyophilized powder (1 mg / rat) or suspension in the dispersing agent described in test example 4 (5 mg / 0.25 ml per rat). Two weeks later, the bone mineral content was determined according to the method described in Test Example 5, and compared with a control group receiving placebo microcapsules. The results are presented in table 7.

 It was found that, regardless of whether the drug was in the form of lyophilized powder or in the form of a suspension, the group with microcapsules containing the drug had a significantly higher content of bone minerals in comparison with the control groups, which indicated the excellent osteogenic activating ability of the pharmaceutical compositions according to this invention.

Test Example 11
A rat model of fibula fracture was obtained according to the method described in Test Example 5. Microcapsules 7 were suspended in a dispersing medium containing various concentrations of sodium phosphate, which was prepared using a solution of D-sorbitol (2.5 g), sodium chloride (0.9 g ) and carboxymethyl cellulose (0.5 g) in distilled water.

 The suspension was administered locally (5 mg / 0.25 ml per rat). Two weeks later, the bone mineral content was determined according to the method described in Test Example 5. The results are shown in Table 7.

The results of the invention
By detecting increased osteogenesis, potentiating activity with low toxicity over an extended period of time, the pharmaceutical composition according to this invention can be safely used as a prophylactic / therapeutic agent for various bone diseases in mammals, for example, bone fractures, osteoporosis, osteomalacia, Behcet syndrome, bone ankylosis spondylitis , rheumatoid arthritis, gonarthritis (gonarthritis deformans) and destruction of joint tissues, as a means of restoring ment of bone tissue after surgery for multiple myeloma, lung cancer, breast cancer, etc. and as an activator of the regeneration of periodontal tissue in case of periodontopathy, etc.

Industrial applicability
A pharmaceutical composition comprising a non-peptidic glazed optically active substance and a lactic acid / glycolic acid copolymer is a valuable agent for the treatment and / or prevention of bone diseases.

Claims (11)

1. Pharmaceutical composition containing non-peptide osteogenesis activating an optically active substance of the formula I

where R ₃ represents a lower alkyl group;
R ₄ and R ₅ independently represent a lower alkyl group or are linked together to form a lower alkylene group,
and a copolymer of lactic acid and glycolic acid. 2. The pharmaceutical composition according to claim 1, in which R ₃ , R ₄ and R ₅ independently represent a C _1-6 alkyl group.  3. The pharmaceutical composition according to claim 1, in which the compound is (2R, 4S) - (-) - N- [4- (diethoxyphosphorylmethyl) phenyl] -1,2,4,5-tetrahydro-4-methyl-7 , 8-methylenedioxy-5-oxo-3-benzothiepin-2-carboxamide.  4. The pharmaceutical composition according to claim 1, wherein the weight ratio of said copolymer to non-peptide osteogenesis of the activating optically active substance is about 1-100-fold.  5. The pharmaceutical composition according to claim 2, which further comprises phosphoric acid or its salt.  6. The pharmaceutical composition according to claim 5, where the phosphoric acid or its salt is sodium phosphate.  7. The pharmaceutical composition according to claim 2, in which the relative content of (2R, 4S) - (-) - N- [4- (diethoxyphosphorylmethyl) phenyl] -1,2,4,5-tetrahydro-4-methyl-7, 8-methylenedioxy-5-oxo-3-benzothiepin-2-carboxamide calculated on the specified copolymer is approximately 5-30 wt.% And the relative content of sodium phosphate based on (2R, 4S) - (-) - N- [4 - (diethoxyphosphorylmethyl) phenyl] -1,2,4,5-tetrahydro-4-methyl-7,8-methylenedioxy-5-oxo-3-benzothiepin-2-carboxamide and said copolymer is about 0.1-20 wt. %  8. The pharmaceutical composition according to claim 1, in which the ratio of lactic acid / glycolic acid is approximately 90 / 10-50 / 50 wt.% And srednevekovoi molecular weight is approximately 8000-50000.  9. The pharmaceutical composition according to claim 1, which is a suspension.  10. The pharmaceutical composition according to claim 1, used for injection.  11. The pharmaceutical composition according to claim 1 for the preparation of agents for the treatment or prevention of bone diseases. Priority on points:
06/05/1995 PP 1-4, 8-11;
06/04/1996 PP 5-7.

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