MXPA06009002A - Sustained release preparation - Google Patents

Abstract

A sustained release preparation which comprises a combination of a microcapsule capable of sustainedly releasing a GnRH agonist or its salt over a long time with another microcapsule capable of sustainedly releasing the GnRH agonist or its salt within a short time.

Description

CONTROLLED RELEASE PREPARATIONS Field of the Invention The present invention relates to a novel controlled release preparation comprising a combination of a microcapsule that gradually releases a GnRH agonist or a salt thereof for a long period and a microcapsule that gradually releases a GnRH agonist or salt. of this for a short period.

Background of the Invention The hormone that releases the luteinizing hormone known as GnRH (or LHRH) is released from the hypothalamus, and inhibits a receptor of the pituitary gland. LH (luteinizing hormone) and FSH (follicle stimulating hormone) that are released by means of this, act on the gonad to synthesize the steroid hormones. However, it has been revealed that, when a compound that has strong hormonal activity releasing luteinizing hormone is released, the number of receptors available decreases and the formation of steroid hormones derived from the gonads is inhibited. Using this, a compound having GnRH activity is applied as a remedy for sex hormone dependent diseases such as prostate cancer, prostatomegaly, endometriosis, Ref .: 174512 hyoid iorna, metrofibroma, precocious puberty, and breast cancer. Such compounds having GnRH activity include, specifically, leuprorelin acetate, buserelin acetate and goserelin acetate, and controlled release preparations containing them. They sell as a remedy for the aforementioned diseases. Preparations containing these compounds have been used initially as a preparation that is administered once a day, after that, they have been formulated in controlled release preparations. Currently, these are sold as a controlled release preparation for one month, three months or four months. These controlled release preparations are disclosed in EP190833 and EP442671. As described above, compounds having GnRH activity are widely used as a remedy for prostate cancer. In diseases that progress slowly, in particular, prostate cancer, the extension of a period of release of a preparation is preferable not only from a point of view of improvement in the therapeutic effect and improvement in the QOL (quality of life) of patients, but also from a medical economics point of view due to the reduction in times of care as a patient who is not hospitalized.

Objective of the Invention However, it is difficult to produce a preparation that exhibits stable controlled release over a long term. In particular, when long-term controlled release is emphasized, an amount of drug is released at an early stage of administration is deficient and the manifestation of the effect of an early stage of administration is released in some cases. In addition, when an amount of drug is released at an early stage of administration is excessive, an amount of the drug release is deficient in a later stage of a sustained release period, and it is difficult to retain the stable controlled release over a long period of time. period in some cases.

Brief Description of the Invention In order to achieve the objective mentioned above, the current inventors studied intensively. Unexpectedly, they found that combining the controlled release preparations of the GnRH agonist having different controlled release terms can increase an amount of the release at an early stage of administration and provide the stable controlled release over a long period. The present inventors conducted other studies based on these discoveries, which originated the termination of the present invention. That is, the present invention is related to. [1] A controlled release preparation comprising a combination of a microcapsule that gradually releases a GnRH agonist or a salt thereof for a long period and a microcapsule that gradually releases a GnRH agonist or a salt thereof for a short period; [2] The preparation according to [1], wherein the GnRH agonist or a salt thereof is a peptide represented by the formula: -oxo-Pro-His-Trp-Ser-Tyr-Y-Leu-Arg-Pro-Z [wherein Y represents a selected residue of DLeu, DAla, DTrp, DSer (tBu), D2Nal and DHis (ImBzl), and Z represents? H-C2H5 or Gly-NH2] or a salt thereof; [3] The preparation according to [1], where the agonist GnRH or a salt thereof is an acetate of a peptide represented by the formula: 5-oxo-Pro-His-Trp-Ser-Tyr-Y-Leu-Arg-Pro-Z; [4] The preparation according to [1], where the long period is 5 months or longer, and the short period is shorter than 5 months; [5] The preparation according to [1], where the long period is 5 months or more and 8 months or less, and the short period is 1 week or longer and shorter than 5 months; [6] The preparations according to [1], wherein the microcapsule is a microcapsule containing a polymer of lactic acid or a polymer of lactic acid-glycolic acid as a base; [7] The preparation according to [1], wherein a combination ratio of the microcapsule that gradually releases a GnRH agonist or a salt thereof for a short period and the microcapsule that gradually releases a GnRH agonist or a - salt of this for a long period is 1: about to 1: approximately 20 expressed as a ratio of a weight of the GnRH agonist or a salt thereof contained in each microcapsule; [8] The preparation according to [1], wherein: the microcapsule that gradually releases the GnRH agonist or a salt thereof for a long period is: a microcapsule containing (i) a GnRH agonist or a salt thereof, and (ii) a lactic acid polymer having a weight average molecular weight of from about 18,000 to about 30,000; and the microcapsule that gradually releases a GnRH agonist or a salt thereof for a short period is: (1) a microcapsule containing (i) a GnRH agonist or a salt thereof, and (ii) a lactic acid-acid polymer glycolic (75/25) (% mol)) having a weight average molecular weight of about 8,000 to about 12,000, or (2) a microcapsule containing (i) a GnRH agonist or a salt thereof, and (ii) ) a lactic acid polymer having a weight average molecular weight of from about 13,000 to about 18,000; [9] The preparation according to [1], wherein: the microcapsule that gradually releases a GnRH agonist or a salt thereof for a long period is: a microcapsule containing (i) a GnRH agonist or a salt thereof, and (ii) a lactic acid polymer having a weight average molecular weight of from about 15,000 to about 50000 wherein a content of a polymer having a weight average molecular weight is 5000 or less is about 5% or minor in weight; and the microcapsule that gradually releases a GnRH agonist or a salt thereof for a short period is: (1) a microcapsule containing (i) a GnRH agonist or a salt thereof, and (ii) a lactic acid-acid polymer glycolic wherein a weight average molecular weight (Mw) is from about 8,000 to about 11,500, and a ratio of a weight average molecular weight (Mw) to a number average molecular weight (Mn) is greater than 1.9, and a molar ratio of the composition of lactic acid to glycolic acid is 99.9 / 0.1 to 60/40, and which does not contain a substance that retains the drug, or (2) a microcapsule that has an order of zero release of an agonist GnRH or a salt thereof for 2 months, and which is prepared by microencapsulating a W / O emulsion prepared with an internal aqueous phase solution containing a GnRH agonist or a salt thereof of about 20 to 70% by weight, and a solution with oil phase that contains, as a a substance controlling the release, a copolymer or a homopolymer wherein a ratio of lactic acid / glycolic acid is from 80/20 to 100/0, and a weight average molecular weight of from about 7,000 to about 30,000; [10] The controlled release preparation according to any of [1] to [9], which gradually releases a GnRH agonist or a salt thereof for a long period; [11] The controlled release preparation according to [10], where the long period is 5 months or longer; [12] An agent for preventing or treating prostate cancer, prostatomegaly, endometriosis, hysteromy, etrofibroma, precocious puberty, dysmenorrhea or breast cancer, or a contraceptive agent, comprising a controlled release preparation as defined in [1]; [13] A process for producing the controlled release preparation as defined in [1], wherein it comprises mixing a microcapsule that gradually releases a GnRH agonist or a salt thereof for a long period and a microcapsule which gradually releases an agonist GnRH or a salt thereof for a short period; [14] A method for preventing or treating prostate cancer, prostatomegaly, endometriosis, hysteromyoma, metrofibroma, precocious puberty, dysmenorrhea or breast cancer, or preventive conception, which comprises administering an effective amount of the controlled release preparation as defined in [1] to a mammal; and [15] The use of a controlled release preparation as defined in [1] to produce a preventive agent or remedy for prostate cancer, prostatomegaly, endometriosis, hysteromyoma, metrofibroma, precocious puberty, dysmenorrhea, or breast cancer. , or contraceptive agent. By combining the microcapsules having different controlled release terms, which gradually release a GnRH agonist or a salt thereof, a preparation can be obtained which has an increasing amount of drug that is released at an early stage of administration and exhibits a release Stable controlled for a long period.

Brief Description of the Figures Fig. 1 is a graph showing the results of Experimental Example 1. • Denotes a blood concentration of peptide A when combo B is administered, and O denotes a blood concentration of peptide A when MC # 2 is administered. Fig. 2 is a graph showing the results of Experimental Example 2. • Denotes a blood concentration of peptide A when combo A is administered, and O denotes a blood concentration of peptide A when administered -MC # 2. Fig. 3 is a graph showing the results of Experimental Example 3. • Denotes a blood concentration of peptide A when combo D is administered, and O denotes a blood concentration of peptide A when MC # 2 is administered.

Detailed Description of the Invention The present invention will be described in detail below. Examples of the GnRH agonist include GnRH agonists which are effective in hormone-dependent diseases, in particular, sex hormone-dependent cancer (eg, prostate cancer, uterine cancer, brecancer, tumor in the pituitary gland etc.), sex hormone-dependent diseases such as prostatomegaly, endometriosis, hysteromyoma, precocious puberty, dysmenorrhea, amenorrhea, premenstrual syndrome, multilocular ovarian syndrome, recurrence to cancer after the operation, dwarfism, Alzheimer's disease , menopausal disorder, indeterminate disease, cancer metsis, and calcium metabolic bone disorder «phosphorus, and contraception (or infertility when the rebound effect is used after cessation of administration). In addition, examples include GnRH agonists that are effective in benign or malignant tumors that are independent of sex hormones but sensitive to GnRH. Examples of this GnRH agonist include, for example, peptides and the like described in Treatment with GnRH analogs, Controversies and Perspectives [published by the Parthenon Publisihing Group Ltd., 1996], JP-A-03-503165, JP-A-03 -101695, JP-A-07-97334 and JP-A-08-259460. As a specific example of the GnRH agonist, a physiologically active peptide is used represented by the general formula [I] 5-oxo-Pro-His-Trp-Ser-Tyr-Y-Leu-Arg-Pro-Z [I] [wherein Y represents a residue selected from a selected residue of DLeu, DAla, DTrp, DSer (tBu), D2Nal and DHis (ImBzl), and Z represents? H-C2H5 or Gly-? H2] or a salt thereof. In particular, a peptide is preferred wherein Y is DLeu and Z is NH-C2Hs or a salt thereof (ie, a peptide represented by 5-oxo-Pro-His-Trp-Ser-Tyr-DLeu-Leu-Arg). -Pro-NH-C2H5 or a salt thereof, in particular, this acetate (leuprorelin acetate: manufactured by Takeda Chemical Industries, Ltd.)). A peptide employed as the GnRH agonist may be the pharmacologically acceptable salt. When the peptide has a basic group such as an amino group, examples of this salt include salts with inorganic acids (eg, hydrochloric acid, sulfuric acid, nitric acid, boric acid, etc.), and organic acids (e.g. eg, carbonic acid, bicarbonic acid, succinic acid, acetic acid, propionic acid, trifluoroacetic acid, etc.). When the peptides have an acid group such as a carboxyl group, examples include salts with inorganic bases (eg, alkali metals such as sodium, potassium, etc., alkaline earth metals such as calcium, magnesium, etc.), and organic bases (e.g., organic amines such as triethylamine, etc., basic amino acids such as arginine, etc.). The peptides can form a complex metal compound (eg, copper complex, zinc complex etc.). These peptides or a salt thereof may be produced by the methods described in the publications mentioned above or gazettes, or methods based thereon. In addition to the leuprorelin mentioned above (leuprorelin acetate), preferred examples of the GnRH agonist include, (1) Goserelin (US-A-4100274, JP-A-52-136172), (2) Bus er ina (USP No. 4,024,248, German Patent No. 2438352, JP-A-51-41359), (3) Triptorelin.

(US-A-4010125, JP-A-52-31073), (4) Naf arel ina US-A-4234571, JP-A-55-164663, JP-A-63-264498, JP-A-64-25794), (5) Histrelin (6) Deslorelin (US-A-4569967, US-A-4218439), (7) Meterelin (PCT WO 91/18016), (8) Gonadrelin (German Patent No. 2213737) and the salts thereof. A microcapsule that gradually releases an agonist GnRH or a salt of this during a short period (from here in "further, simply referred to as" short-period controlled release microcapsule "in some cases), a microcapsule is used that gradually releases a GnRH agonist or a salt thereof for a period of less than 5 months, and uses a microcapsule that gradually releases a GnRH agonist or a salt thereof preferably for a period of about 1 week or more and less than about 5 months, more preferably over a period of about 2 weeks or longer and about 4 months or less, more preferably over a period about 3 weeks or more and about 4 months or less, even more preferably over a period of about 1 month or more and about 3 months or less, more particularly preferred over a period of about 1 month or more and about 2 months or smaller, more preferably for a period of about 1 month to about 3 months, as a microcap If a GnRH agonist or a salt thereof is gradually released over a long period (hereinafter referred to simply as a "long-period controlled release microcapsule" in some cases), a microcapsule is used that gradually releases an agonist GnRH or a salt thereof for a period of 5 months or more, and a microcapsule is used that gradually releases an agonist GnRH or a salt thereof preferably for a period of about 5 months or more and less than 2 years, more preferably over a period of about 5 months or more and one year and 6 months or less, more preferably over a period of 5 months or greater and about 1 year or less, even more preferably during a period of 5 months or more and 8 months or less, is more particularly preferred over a period of 5 months or longer and 6 months or less, more preferably during a period of about 6 months. In the present invention, by "properly combining the" short-period controlled release microcapsule "and the" long period controlled release microcapsule "the microcapsules mentioned above each have a period of continuous release. may combine the use of (i) a microcapsule that gradually releases the GnRH agonist or a salt thereof for about 1 month or about 3 months and (ii) a microcapsule that releases the agonist GnRH or a salt thereof for approximately 6 months. months Specifically, the use of (i) the controlled release microcapsule is described in EP190833 or EP442671 and (ii) the controlled release microcapsule described in WO03 / 002092. As used herein, "combined use" can be the successful administration of a preparation containing the short-period controlled release microcapsule (hereafter, simply referred to as "short-period controlled release preparation" in some cases) and a preparation containing the long-period controlled release microcapsule (hereinafter, simply referred to as "long-period controlled release preparation" in some cases), or the simultaneous administration after mixing the preparation of controlled release of short period and controlled release preparation of long period (in this case, the formulation is included in a preparation after mixing the short-period controlled release microcapsule and the long-period controlled release microcapsule).

Administration of the long-term controlled release preparation after a certain period (eg, after a few hours to a few days) after the administration of the short-period controlled release preparation is also included. However, in the present, "administration" is concerned so that the controlled release periods of the drugs of a short-period controlled release preparation and a long-period controlled release preparation, and the administration of another, are overlapped. preparation after the passage of a period of controlled release of a drug from a preparation is not included in the "combined use" herein. For example, it is not included when a controlled release preparation of one month and a controlled release preparation of three months is used, the case where the controlled release preparation of three months is administered after one month after administration of the drug. controlled release preparation for one month.

A combination ratio of a short-period controlled release preparation and a long-period controlled release preparation is expressed as a weight ratio of a GnRH agonist, usually from 1 to 40 of the long-period controlled release preparation by short-period controlled release preparation, preferably from 5 to 20 of the long-period controlled release preparation by short-period controlled release preparation, more preferably from 7 to 18 (particularly, from 9 to 16) of the release preparation controlled long-period by short-period controlled release preparation, more preferably from 7 to 15 of the long-period controlled release preparation by short-period controlled release preparation, more preferably from 9 to 12 of the controlled release preparation of long period by short-term controlled release preparation (c When mixing a short-period controlled release microcapsule and long-period controlled release microcapsule formulated in a preparation, the combination ratio can be determined by replacing the "short-period controlled release preparation" with "controlled release microcapsule. of short period ", and" preparation of controlled release of long period "by" microcapsule of controlled release of long period ").

The controlled release preparation of the present invention comprises a combination of a long-period controlled release microcapsule and a microcapsule. Short-term controlled release can gradually release a GnRH agonist or a salt thereof for a long period. A long period refers to, for example, 5 months or more, preferably 5 months or more and less than 2 years, more preferably 5 months or more and one year and 6 months or less, more preferably 5 months or more and 1 year or less, more preferably 5 months or more and 8 months or less, more preferably 5 months or more and 6 months or less, more preferably about 6 months. The GnRH agonist, preferably a peptide represented by the formula 5-oxo-Pro-His-Trp-Ser-Tyr-DLeu-Leu-Arg-Pro-NH-C2H5 or a salt thereof (hereinafter, simply referred to as "leuprorelin or a salt thereof" in some cases), more preferably leuprorelin acetate is administered as a controlled release microcapsule, more preferably as an injectable comprising the controlled release microcapsule. The preparation can be produced by combining leuprorelin or a salt thereof, more preferably leuprorelin acetate together with the known pharmaceutically acceptable carrier, excipient, excipient, carrier, antiseptic, stabilizer and binder in the generally acceptable unit dosage form required in the practice of the pharmacy. It is used as an aqueous solution for injection, for example an isotonic solution containing physiological saline, glucose and other auxiliary agents (eg, D-sorbitol, D-mannitol, sodium chloride). An appropriate solubility agent such as alcohol can be used together (eg, ethanol), polyalcohol (eg, - propylene glycol, polyethylene glycol), and a non-ionic surfactant (eg, Polysorbate 80 (TM), HCO-50). As an oily solution, for example, sesame oil and soybean oil are used, and solubilizing agent such as benzyl benzoate and benzyl alcohol can be used together. The preparation can be mixed with, for example, a buffer, sodium acetate buffer), a sedative agent (eg, benzalkonium chloride, procaine chloride etc.), a stabilizer (eg, serum albumin). of human, polyethylene glycol, etc.), a preservative (eg, benzyl alcohol, phenol, etc.), and an antioxidant. The prepared injectable solution is usually filled in an appropriately sealed container such as a vial and a vial. Specifically, a controlled release preparation (particularly, controlled release microcapsule) containing the GnRH agonist (preferably leuprorelin or a salt thereof, more preferably leuprorelin acetate) can be produced according to methods known per se, for example, from according to the method described in EP190833, EP442671, and WO03 / 002091 for a short-period controlled release preparation (or short-period controlled release microcapsule), or according to the method described in WO03 / 002092, for a release preparation long period controlled (or long period controlled release microcapsule). The preparation of the present invention can be produced by mixing a microcapsule which generally releases a GnRH agonist or a salt thereof for a long period with a microcapsule which gradually releases a GnRH agonist or a salt thereof for a short period. That is, a controlled release preparation of the present invention comprising a combination of a microcapsule that gradually releases a GnRH agonist or a salt thereof for a long period and a microcapsule that gradually releases a GnRH agonist or a salt thereof during a period of time. short period can be prepared separately by producing a microcapsule that gradually releases a GnRH agonist or a salt thereof for a long period and a microcapsule that gradually releases the GnRH agonist or a salt thereof for a short period, the appropriate mixture with a ratio of proper mixing, and formulating the mixture. The mixture of a long-period controlled release microcapsule and a short-period controlled release microcapsule can be developed before or after a primary drying step and a secondary drying step described below. However, when administered separately with an interval, it is not necessary to mix a microcapsule that gradually releases a GnRH agonist or a salt thereof for a long period and a microcapsule that gradually releases a GnRH agonist or a salt thereof during a period of time. short period, and each microcapsule can be formulated in a preparation for administration before each use. Alternatively, two controlled release preparations comprising short-period and long-period controlled release microcapsules, respectively, can be mixed in the administration. An example of a process for producing the controlled release microcapsule will be described below. First, a GnRH agonist (preferably leuprorelin or a salt thereof, more preferably acetate) is dissolved in water at about 20 to 70% (w / w), preferably 25 to 65% (w / w), more preferably at 35 to 60% (w / w), and if necessary, a substance that retains the drug such as gelatin and basic amino acid is dissolved or suspended in it, to obtain a solution with internal aqueous phase. To the internal aqueous phase solution can be added carbonic acid, acetic acid, oxalic acid, citric acid, phosphoric acid, hydrochloric acid, sodium hydroxide, arginine, lysine or a salt thereof as a pH adjusting agent to retain the stability and solubility of a GnRH agonist (preferably leuprorelin or a salt thereof), more preferably leuprolein acetate). In addition, albumin, gelatin, citric acid, sodium ethylenediamine tetraacetate, dextrin, sodium bisulfite, and polyol compound such as polyethylene glycol can be added as a stabilizer for a GnRH agonist (preferably leuprorelin or a salt thereof, more preferably sodium acetate). leuprorelin, or the esters of para-oxybenzoic acid (methylparaben, propylparaben, etc.), or benzyl alcohol, chlorobutanol, and thimerosal-as a preservative are generally used.

In the internal aqueous phase solution obtained in this way, it is added to a solution (oil phase) containing a high molecular weight polymer (polymer), and an emulsion process was carried out to make an emulsion of the W / O type. As the emulsion process, the known dispersion methods are used, for example, intermittent vibration method, method using a mixer such as a stirrer of the propellant type and a stirrer of the turbine type, colloidal milling method, homogenizing method, and ultrasound irradiation method. Then, the W / O type method prepared in this way is subjected to a microemulsion step, where the water drying method or phase separation method can be applied. When a microcapsule is produced by the water drying method, the W / O emulsion is added to a third aqueous phase to form a three phase emulsion of the W / O / W type, then a solvent is evaporated in an oil phase to prepare a microcapsule. An emulsifier can be added to the aqueous phase of the external phase. For example, it can generally be any emulsifier until a stable O / W type emulsion is formed, and examples include an anionic surfactant (sodium oleate, sodium stearate, sodium lauryl sulfate, etc.), an ionic surfactant (ester) of polyoxyethylene sorbitan fatty acid [Tween 80, Tween 60, Powder Back], polyoxyethylene ricino oil derivative [HCO-60, HCO-50, Nikko Chemicals Co., Ltd.]), and polyvinylpyrrolidone, polyvinyl alcohol, carboxymethylcellulose, lecithin and gelatin. One of these can be used alone, or some of these can be used in combination. A concentration of the emulsifier with use can be appropriately selected from a range of about 0.01% to 20%, more preferably, about 0.05 to 10%. To evaporate a solvent from an oil phase, a method is generally adopted. In the method, evaporation is developed by gradual evacuation while stirring with a stirrer of the propellant type of a magnetic stirrer, or by regulating the degree of vacuum using a rotary evaporator. In this case, while the solidification of a high molecular weight polymer is carried out to a certain degree, an emulsion of the type W / 0- / W is gradually made with the purpose of making the dropout of a solvent completely possible. shorten the necessary time. The microcapsule thus obtained is separated by centrifugation or filtration, washed with distilled water several times a free GnRH agonist (preferably leuprorelin or a salt thereof, more preferably leuprorelin acetate), a drug that retains the substance and an adhering emulsifier. to a surface of a microcapsule, they are dispersed again in distilled water and lyophilized. After this, an agent which retains the aggregation such as sugar is added. Water and an organic solvent are adsorbed in a microcapsule under reduced pressure with heating, if necessary. In lyophilization, specifically, sugar is added to the microcapsule thus obtained from about 2 to about 60% by weight relative to the microcapsule, followed by lyophilization. This step is called the primary drying step. The material is then heated appropriately in a range from the glass transition temperature of a polymer to a temperature of about 40 ° C higher than the glass transition temperature. This step is called the secondary drying step. Example of the sugar to be used includes D-mannitol, sodium alginate, fructose, dextran, dextrin, sucrose, D-sorbitol, lactose, glucose, maltose, starches, and trehalose. These sugars can be used alone, or can be used appropriately when mixed. Among these, D-mannitol is particularly preferred, which is freeze-dried easily and has little toxicity. The method for adding sugar includes, but is not limited to, dispersing a microcapsule well in an aqueous sugar solution, and simply adding to the microcapsule before mixing them with a mixer., etc. An amount of a sugar to be added is preferably about 5 to about 40% by weight relative to a microcapsule. When the microcapsule has already been mixed with the sugar, for example sugar is used in or before the water is dried or spray dried and then mixed, sugar can be added in view of the amount of this so that the total amount is present. within the range mentioned above. The lyophilization can be carried out according to the known method. A heating temperature of the secondary drying step is preferably in a range from the glass transition temperature of a polymer to a temperature of 20 ° C higher than the glass transition temperature. The heating temperature is selected so that the temperature of a product is generally in a range of about 30 to about 60 ° C. Here, a vitreous transition temperature refers to an intermediate point of the glass transition temperature obtained when it is raised to a heating rate of 10 or 20 ° C per minute using a differential scanning calorimeter (DSC). The heating time of the secondary drying step is not particularly limited, but is usually from about 1 to about 240 hours, preferably about 10 to about 120 hours, more preferably about 20 to about 72 hours. The heating temperature, the heating time, the degree of drying, and the heating method are determined by the diameter of the particle, the stability, glass transition temperature, the melting point, the ease of deformation of the microcapsule , the stability of a drug contained in it, the type and amount of sugar added to it, and a degree of dispersion of the microcapsule. By means of this heating, the water and the organic solvent in the microcapsule can be completely eliminated. When the microcapsule is produced by means of a phase separation method, a coacervating agent is gradually added to the W / O emulsion under stirring to precipitate and molt a high molecular weight polymer. The coacervating agent may be of the polymer type, of the mineral oil or vegetable oil type type which is compatible with a solvent of the high molecular weight polymer and does not dissolve an encapsulating polymer. Such coacervating agent includes silicone oil, sesame oil, soybean oil, corn oil, cottonseed oil, coconut oil, linseed oil, mineral oil, n-hexane, and n-heptane. These can be used-when mixing two or more types. - - The microcapsule thus obtained is collected by filtration, washed with heptane repeatedly to eliminate the coacervating agent. Then, by the same method as the water drying method, a free drug is removed, and the solvent is resorbed. In order to prevent aggregation of the particles during washing, an agent can be added to prevent aggregation. The microcapsule obtained above is milled slightly as necessary, passed through a screen to remove a part of the microcapsule that is too large. A diameter of a microcapsule is in a range of about 0.5 to 1000 μm, more preferably it is desired that the particle diameter be in the range of about 2 to 500 μm. When used as an injectable suspension, the particle diameter is in a range that the dispersion and the needle penetration property are satisfied, it is desirable that the particle diameter is, for example, in a range of about 2 to about 100 μm. As the high molecular weight polymer, a biodegradable polymer is used, for example, a polymer, a copolymer or a mixture thereof which is synthesized from one or more types of α-hydroxycarboxylic acid such as α-hydroxymonocarboxylic acid ( e.g., glycolic acid, lactic acid, etc.), α-hydroxydicarboxylic acid (eg, malic acid), and α-hydroxytricarboxylic acid (eg, citric acid) and has a free carboxyl group; poly (α-cyanoacrylic acid ester); polyamino acid (e.g., poly (β-benzyl-L-glutamic acid), etc.); maleic anhydride copolymer (e.g., maleic acid-styrene copolymer, etc.).

The monomer linkage form can be either random, block and grafted. When the α-hydroxymonocarboxylic acid, α-hydroxydicarboxylic acid or α-hydroxytrricarboxylic acid has an optical active center in a molecule, any of the D-, L- and DL- isomers can be used. Among these, a lactic acid-glycolic acid polymer (hereinafter referred to as poly (lactide-co-glycolide), poly (lactic acid-co-glycolic acid) or a copolymer of lactic acid-glycolic acid in some In addition, unless otherwise indicated, the homopolymer (polymer) and copolymer of lactic acid and glycolic acid are collectively called in. In addition, the homopolymer of lactic acid called lactic acid polymer, polylactic acid or polylactide is preferred, and glycolic acid homopolymer called glycolic acid polymer, polyglycolic acid, or polyglycolide in some cases) and poly (a-cyanoacrylic acid). More preferable is a polymer of lactic acid glycolic acid, and more preferably it is a polymer of lactic acid glycolic acid having a free carboxyl group at the end.A biodegradable polymer can be a salt Examples of the salt include salts with inorganic bases (e.g., alkali metal such as sodium and potassium, alkaline earth metals such as calcium and magnesium) and organic bases (e.g., organic amines such as triethylamine, basic amino acids such as arginine), and salts and salts of complexes with transition metals (eg, zinc, iron, copper, etc.) When the lactic acid-glycolic acid polymer is used as a biodegradable polymer, its composition ratio (% mol ) is preferably about 100/0 to about 40/60, more preferably about 100/0 to about 50/50 In the case of the microcapsule with controlled release that releases the order of zero agonist GnRH for 2 months, a lactic acid homopolymer (lactic acid polymer) having a composition ratio of 100/0 is also preferably used. An optical isomer ratio of lactic acid which is one of the minimum repeating units of the "lactic acid-glycolic acid polymer" is preferably such that the D-isomer / L-isomer (% mol / mol) is in the range from about 75/25 to about 25/75"." By this the polymer D-isomer / L-isomer is generally used. { % mol / mol), in particular, the polymer having a range of about 60/40 to about 30/70. A weight average molecular weight of the "lactic acid-glycolic acid polymer" or "lactic acid polymer" is generally about 3,000 to about 100,000, preferably about 3,000 to about 60,000, more preferably about 3,000 to about 50,000. In the present invention, for example, a preparation using a lactic acid-glycolic acid polymer (75/25 (mol%)) with a weight-average molecular weight of 8,000 to 12,000 or a polymer of lactic acid having a weight average molecular weight of 13,000 to 18,000 as a base for a controlled release microcapsule with a short period, and a preparation using a lactic acid polymer with a weight average molecular weight of 18,000 to 30,000 as a base for a microcapsule controlled release with long period. Dispersion (weight average molecular weight / number average molecular weight) usually is preferably about 1.2 to about 4.0, particularly preferably "about 1.5 to about 3.5, an amount of a free carboxyl group of the lactic acid-glycolic acid "or" lactic acid polymer "is usually preferably about 20 to about 1000 μmol (micromoles), preferably about 40 to about 1000 μmol (micromoles) per unit mass (grams) The weight average molecular weight, number average molecular weight and dispersion are molecular weights in terms of polystyrene as measured by gel permeation chromatography (GPC) using polystyrenes having molecular weights known as the standard substance., the dispersion. The standard substance includes, for example, the following combinations: (1) polystyrenes having known average molecular weight weights of about 500, about 1,000, about 3,000, about 5,000, about 10,000, about 20,000, about 50, 000 and about 100,000, (hereinafter standard substance A) (2) 10 types of polystyrenes having known average molecular weight weights of about 500, about 1,000, about 2,500, about 5,000, about 10,000 , about 20,000, about 50,000, about 100,000, about 200,000 and about 400,000 (hereinafter, standard substance B) (3) 8 types of polystyrenes having known average molecular weights of 98900, 37200, 17100 , 9490, 5870, 2500, 1051 and 495 (from hereinafter, standard substance C). The measurement is developed by using a GPC device (HLC-8020GPC manufactured by Tosoh Corporation; a detector is a differential refractometer) and a GPC column (manufactured by Tosoh Corporation, a column where TSK gel G4000HHR, the TSK gel G3000HHR, the TSKgel G2000HHR and TSKgel G1000HHR connect from a sample inlet into a order from a higher exclusion limit), and using tetrahydrofuran as the mobile phase. The flow rate is 1.0 ml / min. The amount of the above free carboxyl group refers to that which is obtained by the labeling method (hereinafter referred to as the "amount of carboxyl group by the labeling method"). Specifically, the case of polylactic acid is as follows. W mg of polylactic acid is dissolved in 2 ml of 5N hydrochloric acid / acetonitrile solution (v / v = 4/96), 2 ml of 0.O1 M o-nitrophenylhydrazine hydrochloride (ONPH) solution (acid) hydrochloric acid / acetonitrile / ethanol = 1.02 / 35/15 5N), 2 ml of solution hydrochloride l-ethyl-3- (3- dimethylaminopropyl) -carbodimida (pyridine / ethanol = 4v / 96v) and reacted at 40 ° C for 30 minutes, and the solvent was distilled off. The residue was washed with water (4x), dissolved in 2 ml of acetonitrile, and 1 ml of potassium hydroxide ethanolic solution of 0.5 mol / L was added and reacted at 60 ° C for 30 minutes. The reaction solution was diluted with 1.5N aqueous sodium hydroxide solution to Y ml, and 544 nm of absorbance A (/ cm) was measured using an aqueous solution of sodium hydroxide or a control. On the other hand, when an aqueous solution of DL-lactic acid was used as a standard substance, an amount of its free carboxyl group Cmol / L was obtained by alkaline titration, and 544 nm of absorbance of the lactic acid hydrazide can be obtained in The O? PH labeling method is B (/ cm), a molar amount of the free carboxyl group per unit mass (grams) of a polymer by means of the following equation: [COOH] (mol / g) = (AYC) / (WB) Alternatively, although the "carboxyl group amount" is also obtained by dissolving the biodegradable polymer in a mixed solvent of toluene-acetone-methanol, and titrating the carboxyl group in this solution with an alcoholic solution of potassium hydroxide using phenoftalein as an indicator (hereinafter, a value obtained by this method is referred to as the "amount of carboxyl group by the alkaline titration method"). However, the reaction competes with the hydrolysis of the polyester of the main chain during the titration, as a result, there is a possibility that the endpoint of the titration is not so clear, and therefore it is desirable to determine by the labeling method . The "lactic acid-glycolic acid polymer" or "lactic acid polymer" can be produced, for example by polycondensation by dehydration without catalyst of lactic acid and glycolic acid, or lactic acid (JP-A-61 28251) or polymerization with opening the ring using a catalyst of a cyclic diester compound such as lactide, and glycolide, or lactide (Encyclopedic Handbook of Biomaterials and Bioengineering Part A: Materials, Volume 2, Marcel Dekker, Inc., 1995). The polymer obtained by the known ring opening polymerization method does not necessarily have a free carboxyl group at the end of the resulting polymer, but by subjecting to a hydrolyzing reaction described in EP-A-0839525, it can be modified into a polymer that it has a degree of an amount of a carboxyl group per unit mass, and this too - can be used. The "lactic acid-glycolic acid polymer having a free carboxyl group at the end" or "lactic acid polymer having a free carboxyl group at the end" above can be produced by the same process as the known process (e.g. , see the method of polycondensation with dehydration without catalyst, of JP-A-61-28521, JP-A-10-182496, JP-A-2000-234016) or a process Vimilar. More specifically, as a long-period controlled release microcapsule, a microcapsule (A) - described, for example, in WO 03/002092, containing (i) a GnRH agonist or a salt thereof and (ii) a polymer of Lactic acid having a weight average molecular weight of about 1500 to about 50000 where a content of a polymer having a weight average molecular weight of not more than 5000 is used is not more than 5% by weight. In the microcapsule (A), a content of a GnRH agonist or a salt thereof is, for example, about 0.001 to about 50% (w / w), preferably about 0.02 to about 40% (w / w), more preferably about 0.1 to about 30% (w / w), more preferably about 0.1 to about 24% (w / w), particularly preferably about 3 to about 24% (w / w), more preferably around 14 to about 24% (w / w) in relation to the entire preparation.

A lactic acid polymer is preferably a polymer wherein a content of a polymer having a molecular weight of not more than 5000 is not more than about % by weight and a content of a polymer having a molecular weight of not more than 3000 is not more than 1.5% by weight, more preferably a polymer wherein the content of a polymer having a molecular weight not greater than 5000 is not greater than 5% by weight, a content of a polymer having a molecular weight of not more than 3000 is not greater than about 1.5% by weight, and a content of a polymer having a molecular weight not greater than 1000 is not greater than about 0.1% by weight. A weight average molecular weight of a lactic acid polymer is preferably 15,000 to 40000, more preferably about 15,000 to about 30,000, more preferably about 17,000 to about 30,000. The weight average molecular weight in this case can be determined, for example, using the standard substance B. A short-period controlled release microcapsule to be used for this, for example, (1) a microcapsule (B) containing (i) a GnRH agonist or a salt thereof, and (ii) a lactic acid-glycolic acid polymer wherein a weight average molecular weight (Mw) is from about 8,000 to about 11,500, and a ratio of a weight average molecular weight (Mw) to an average molecular weight in number (Mn) is greater than 1.9, and a molar ratio of the composition of lactic acid to glycolic acid is 99.9 / 0.1 to 60/40, and which does not contain a substance that retains the drug, or (2) a microcapsule described in EP44267Í (C) having an order of zero release of a GnRH agonist or a salt thereof for 2 months, and which is prepared by microencapsulating a W / O emulsion prepared with an internal aqueous phase solution containing a GnRH agonist or a salt of this in approximate 20 to 70% by weight, and an oil phase solution containing, as a substance controlling the release, a copolymer or a homopolymer wherein a ratio of lactic acid / glycolic acid is 80/20 to 100/0, and the weight average molecular weight is from about 7,000 to about 30,000. The microcapsule (B) is a novel microcapsule, preferably, it is a microcapsule produced by mixing (i) a solution containing a GnRH agonist or a salt thereof and does not contain a substance that retains the drug and (ii) a regulated solution. about 25 to about 35 ° C, which contains a polymer of lactic acid-glycolic acid or a salt thereof where a weight average molecular weight (Mw) is from about 8,000 to about 11,500, a ratio of the weight average molecular weight (Mw) to the number average molecular weight (Mn) is greater than 1.9, and a molar ratio of the lactic acid to acid composition glycolic is 99.9 / 0.1 to 60/40 to produce an emulsion of the W / O type at about 25 to about 35 ° C, cooling this emulsion to about 15 about 20 ° C, dispersing the emulsion of the W / O type at an aqueous phase to produce an emulsion of the W / O / W type with the drying of water. The lactic acid-glycolic acid polymer or a salt thereof is used in the microcapsule (B) is a polymer of lactic acid-glycolic acid or a salt thereof where the weight average molecular weight (Mw) is about 8,000 about 11,500, a ratio of the weight average molecular weight (Mw) of the lactic acid-glycolic acid polymer to the number average molecular weight (Mn) of the lactic acid-glycolic acid polymer is greater than 1.9, a molar ratio of the composition of lactic acid to glycolic acid is 99.9 / 0.1 to 60/40. Examples of the salt of the lactic acid-glycolic acid polymer include salts with inorganic bases (eg, alkali metal such as sodium and potassium, alkaline earth metals such as calcium and magnesium) and organic bases (eg, organic amines such as triethylamine, basic amino acids such as arginine), and salts and complex salts with transition metals (eg, zinc, iron, copper, etc.). A ratio (Mw / Mn) weight average molecular weight (Mw) of the lactic acid-glycolic acid polymer with the number average molecular weight (Mn) of the lactic acid-glycolic acid polymer is preferably about 1.95 to about 4.0, about 2.0 to about 3.5, more preferably about 2.3 to about 3.1. A compositional ratio (% mol) of the lactic acid-glycolic acid polymer is preferably 99/1 to 60/40, more preferably 90/10 to 60/40, more preferably 80/20 to 60/40, more particularly preferably 80/20 to 70/30, inter alia, 75/25 is preferred. The weight average molecular weight of the lactic acid-glycolic acid polymer is usually about 8,000 to about 11,500, preferably about 9,000 to about 11,500, more preferably about 9,500 to approximately 11,000. As used herein, a weight average molecular weight, a number average molecular weight, and the dispersion refers to a molecular weight (weight average and number average) in terms of polystyrene as measured by .gel permeation chromatography. (GPC) using a few types of polyesters having weight average molecular weights as the standard substance, and the dispersion was calculated. A column and a mobile phase can be appropriately selected in the measurement. A copolymer of lactic acid-glycolic acid is dissolved in dichloromethane, water is added and divided. The number-average molecular weight can be calculated by titrating a layer of dichloromethane with an ethanolic solution of potassium hydroxide using an automatic titration device, and calculating a terminal carboxylic acid amount. Thereafter, this is expressed as a number-average molecular weight by the quantification of the terminal group. An average molecular weight in number by the quantification of the terminal group is an absolute value, while a number average molecular weight per measurement with GPC is a relative value that varies depending on the test or analysis conditions (eg, a type of mobile phase, a type of column, a standard substance, the selection of the width of sliding a baseline, etc.), therefore, it is difficult to digitize it primarily. However, for example, in a polymer synthesized from lactic acid and glycolic acid by the method of polycondensation with dehydration without catalyst and having a free carboxyl group at the end, a number average molecular weight measured by GPC and an average molecular weight in number by quantification of the terminal group are approximately consistent. In the case of this glycolic acid polymer, approximately consistent means that a number average molecular weight by quantification of the terminal group is in a range of about 0.2 to about 1.5-fold the number average molecular weight measured by GPC , preferably in the range of about 0.3 to about 1.2-fold. A GPC method of Reference Example 5 is a method GPC measuring using 8 types of standard polystyrene products (standard substance C) having a weight average molecular weight (Mw) of 98900, 37200, 17100, 9490, 5870, 2500, 1051 and 495 evaluated by a GPC method. The weight average molecular weight and number average molecular weight of the lactic acid-glycolic acid polymer or a salt thereof is used in the microcapsule (B) can be measured, for example, using a GPC method of Reference Example 5. More specifically, the following lactic acid-glycolic acid polymers are preferably used: (1) copolymer of lactic acid-glycolic acid (lactic acid / glycolic acid = 75/25, Mw = approximately 10300, Mn = approximately 4000, ratio of Mw / Mn = 2.6 (one value by the GPC method (old method) of Reference Example 5)) (2) copolymer of lactic acid-glycolic acid (lactic acid / glycolic acid = 75/25, Mw = approximately 10,400, Mn) = approximately 4100, ratio of Mw / Mn = 2.5 (a ppr value for the GPC method (old method) of the Reference Example )) 'Although a rate of degradation and / or elimination of the lactic acid-glycolic acid polymer varies greatly depending on the composition or molecular weight of the polymer. Generally, while decreasing the glycolic acid fraction, "the degradation and / or elimination is slower, therefore, a release period can be extended by reducing the glycolic acid fraction or by increasing the molecular weight. The release can be shortened by increasing the glycolic acid fraction, or by reducing the molecular weight.In order to obtain a controlled release preparation of the long period type (eg, 1 to 12 months, preferably 1 to 6 months), it prefers a lactic acid-glycolic acid polymer having a composition ratio and a weight-average molecular weight in the aforementioned range When a lactic acid-glycolic acid polymer is selected that degrades more rapidly than the lactic acid polymer -glycolic acid having the composition ratio and the weight average molecular weight in the aforementioned range, it is difficult to control ol of liberation at an early stage.

Conversely, when the lactic acid-glycolic acid polymer is selected that degrades more slowly than the "lactic acid-glycolic acid" polymer having the composition ratio and the weight-average molecular weight in the aforementioned range, it is easily generated a period during which an effective amount of a drug is not released A lactic acid-glycolic acid polymer can be produced, for example, by polycondensation with dehydration without lactic acid catalyst and glycolic acid (JP-A-61- 28521) or polymerization with ring opening using a catalyst in a cyclic form such as lactide and glycolide (Encyclopedic Handbook of Biomaterials and Bioengineering Part A: Materials, Volume 2, Marcel Dekker, Inc., 1995). Although a polymer is synthesized by ring-opening polymerization it is a polymer having no carboxyl group, a polymer can also be used where it is ultimately converted to a free carboxyl group by chemical treatment of the polymer (J. Controlled-Release, Vol. 41, pp. 249-257, 1996). The lactic acid-glycolic acid polymer having a carboxyl group at the end can be produced by the known process (for example, polycondensation method with dehydration without catalyst, see JP-A-61-28521) without any problem and, in addition , a polymer having a free carboxyl group at the site can be produced, not limited to its terminus by the known process (for example, see W094 / 15587). "" While the lactic acid-glycolic acid polymer from which it has ultimately been converted to a free carboxyl group by chemical treatment after ring-opening polymerization, those which are commercially available, for example, from Boehringer, can be used.

Ingelheim KG. In addition, the hydrolysis of the lactic acid-glycolic acid polymer produced by polymerization with ring opening in the presence of an acid or a base is developed according to the known method. In addition, hydrolysis is developed in the presence of water. Here, the acid includes inorganic acids such as hydrochloric acid, nitric acid, sulfuric acid and phosphoric acid, and organic acids such as lactic acid, acetic acid, tartaric acid, citric acid and succinic acid. The base includes alkali metal hydroxide such as sodium hydroxide and potassium hydroxide, and alkali metal carbonate such as sodium carbonate and potassium carbonate. When hydrolysis is developed in the presence of a base, the release of a GnRH agonist or a salt thereof from the controlled release microcapsule is affected by a remaining amount of the base. Therefore, it is preferable to develop the hydrolysis in the presence of an acid. - - Hydrolysis generally develops in a solvent that has no adverse effect on the reaction. Such a solvent includes alcohols such as methanol, ethanol and propanol, ethers such as tetrahydrofuran, dioxane, diethyl ether and diisopropyl ether, water and a mixture of solvents thereof. Alternatively, an excessive amount of the above acid or base can be used as a solvent. A temperature of the hydrolysis is, for example, from about 0 to about 100 ° C, preferably about 10 to about 100 ° C. Since the time required for hydrolysis is different depending on the weight average molecular weight of the polyacetic acid produced by ring opening polymerization, the type of acid or base, the type of solvent, and the temperature, the time may properly determined by collecting a portion of the lactic acid-glycolic acid polymer during the hydrolysis, and measuring the weight average molecular weight of the lactic acid-glycolic acid polymer. The time required "for the hydrolysis is not particularly limited, but is for example, about 1 hour to about 10 days, preferably about 10 hours to about 5 days.The lactic acid-glycolic acid polymer produced by the polymerization with opening of ring can produce only a controlled release microcapsule having great initial release, but in a polymer of lactic acid-hydrolyzed glycolic acid, that is, the lactic acid-glycolic acid polymer used in the present invention, can produce a microcapsule of sustained release having small initial release It is preferable that the lactic acid-glycolic acid polymer is further subjected to a purification step.A purification step is carried out by dissolving the lactic acid-hydrolyzed glycolic acid polymer, emptying the solution resulting in water or a mixed solution of water and a solvent organic soluble in water, and separating by precipitation the lactic acid-glycolic acid polymer. The organic solvent includes, for example, halogenated hydrocarbons (eg, dichloromethane, chloroform, chloroethane, dichloroethane, trichloroethane, carbon tetrachloride.), Ketones (eg, ketones, etc.), ethers (e.g. ., tetrahydrofuran, ethyl ether, isopropyl ether, etc.), esters (e.g., ethyl acetate, butyl acetate, etc.), and aromatic hydrocarbons (e.g., benzene, toluene, xylene, etc.). ). An amount of the organic solvent used is, for example, about 3 to about 20 times (w / v) relative to the hydrolyzed polylactic acid. The water-soluble organic solvent includes, for example, acetone, methanol, ethanol, tetrahydrofuran and acetonitrile. An amount of water or a mixed solution of water and a water-soluble organic solvent to be used is not particularly limited, but is generally an excessive amount greater than hydrolysed polylactic acid. A temperature in a purification step is usually around 0 to about 90 ° C, preferably around 20 to about 70 ° C. In the purification step described above, water-soluble low molecular weight compounds are removed (e.g., compounds having a weight average molecular weight of about 1,000 or less). When a lactic acid-glycolic acid polymer obtained by means of the purification step is used, the extraction ratio (trap ratio) of a GnRH agonist or a salt thereof can be improved with the production of a controlled release microcapsule, and a controlled release preparation having a strong reduced initial release can be produced. Further, bringing the lactic acid-glycolic acid polymer produced by the ring-opening polymerization to hydrolysis and the purification step allows to produce the lactic acid-glycolic acid polymer substantially free of a harmful catalyst (e.g. of zinc, such as zinc oxide and tin compound such as stannous octanoate) used in polymerization with ring opening. A substance that retains drug is a substance that has a characteristic that is soluble in water but sparingly soluble in an organic solvent in an oily phase, it becomes a semi-solid that easily becomes highly viscous in the state where it dissolves in water , or the viscosity is markedly increased by some extrinsic factor such as a temperature, a pH, a metal ion (e.g., Cu2 +, Al3 +, Zn +, etc.), an organic acid (e.g., tartaric acid, citric acid, tannic acid, etc.), or a salt thereof, and a gum condensation agent (eg, gutaraldehyde, acetoaldehyde, etc.), to form a semisolid or solid matrix, as an example of the substance which retains the drug, natural or synthetic gums or high molecular weight compounds are used.The natural gum includes acacia gum, gum arabic, Irish moss, Baraya gum, tragacanth gum, guayaca gum, xanthan gum, and seed gum of the white acacia. Natural high molecular weight composition includes a protein such as casin, gelatin, collagen, albumin (e.g., human albumin serum), globulin and fibrin, and a carbohydrate such as cellulose, dextrin, pectin, starch, agar agar, and tomorrow. These may be as they are, or they may be a partially chemically modified synthetic gum, for example, the above-mentioned natural gum which has been esterified or etherified (e.g., methylcellulose, ethylcellulose, carboxymethylcellulose, gelatin succinate, etc.). ), or hydrolyzed (eg, sodium alginate, sodium pectinate etc.), or a salt thereof. The high molecular weight synthetic compound includes, for example, polyvinyl compound (eg, polyvinylpyrrolidone, polyvinyl alcohol, polyvinyl methyl ether, polyvinyl ether, etc.), polycarboxylic acid (eg, polyacrylic acid, acid. polymethacrylic, Carbopol, (Goodrich), etc.), a polyethylene compound (e.g., polyethylene glycol, etc.), a polysaccharide (e.g., polysucrose, polyglucose, polylactose, etc.) and a salt thereof . In addition, this substance that can proceed in the condensation or cross-linking by the extrinsic factor mentioned above originates a high molecular weight compound is also included. Among these compounds, inter alia, gelatin, albumin, pectin or agar-agar, in particular, gelatin compounds to these.

The microcapsule (B) may contain microparticles (which is a microsphere) containing a GnRH agonist or a salt thereof and a lactic acid-glycolic acid polymer or a salt thereof. Examples of the microparticle include a microcapsule containing a core of a GnRH agonist or a salt thereof in a - particle, a polynuclear microcapsule containing many nuclei of a GnRH agonist or a salt thereof in a particle, and a microparticle in wherein a GnRH agonist of molecular form or a salt thereof is dissolved or dispersed in a polymer of lactic acid-crude glycolic acid as a solid solution. The content of a GnRH agonist, or a salt thereof in the microcapsule (B) is different depending on the type of GnRH agonist or a salt thereof, the desired pharmacological effect, and the duration of the effect, and is for example about 0.1. at about 50% (w / w), preferably around 0.1 to about 30% (w / w), preferably around 5 to about 24% (w / w). A process for producing the microcapsule (B) will be described in detail below. The microcapsule (B) is produced by mixing (i) a solution containing a GnRH agonist or a salt thereof, and does not contain a substance that retains the drug and (ii) a solution, regulated at about 25 to about 35 ° C, which contains a polymer of lactic acid-glycolic acid or a salt thereof (hereinafter, abbreviated as biodegradable polymer) wherein the weight average molecular weight (Mw) is about 8,000 to about 11,500, a ratio of the weight average molecular weight (Mw) and the number average molecular weight (Mn) is greater than 1.9, and a molar ratio of the composition of lactic acid to glycolic acid is from 99.9 / 0.1 to 60/40, to produce a W / O type emulsion at about 25 to about 35 ° C (primary emulsion), cooling this emulsion to about 15 about 20 ° C, dispersing the W / O type emulsion in an aqueous phase to produce an W / O / W type emulsion (secondary emulsion), and holding the emulsion of the emulsion. Type W / O / W to water drying. An W / O type emulsion containing a solution containing a GnRH agonist or a salt thereof, and does not contain a substance that retains the drug as an internal aqueous phase, and a solution, regulated at about 25 to about 35 ° C, which contains a biodegradable polymer as an oil phase can be produced as follows. First, a GnRH agonist or a salt thereof is dissolved in water (preferably distilled water for injection) at a concentration of about 0.001 to about 90% (w / w), preferably about 0.01 to about 80% (w / w) ) more preferably around 1 to about 70% (w / w), particularly preferably about 50%, to form an internal aqueous phase. To the internal aqueous phase can be added carbonic acid, acetic acid, oxalic acid, citric acid, phosphoric acid, hydrochloric acid, sodium hydroxide, arginine, lysine, and a salt thereof as a pH adjusting agent to maintain stability and solubility of a GnRH agonist or a salt thereof. In addition, albumin, gelatin, trehalose, citric acid, ethylenediamine sodium tetraacetate, dextrin, cyclodextrin can be added as a stabilizer. (a-, β-, β- and a derivative thereof (eg, maltosyl β-cyclodextrin, β-cyclodextrin sulfobutyl ether, etc.), sodium bisulfite, a polyol compound such as polyethylene glycol, acid ester polyoxyethylene sorbitan fatty acid [eg, Tween 80, Tween 60, (Kao Corporation, Japan)] a surfactant such as polyoxyethylene ricino oil derivative [eg, HCO-60, HCO-70, (Nikko Chemicals Co., Ltd.]), esters of para-oxybenzoic acid (eg, methylparaben, propylparaben, etc.), benzyl alcohol, chlorobuthanol, and thimerosal for a GnRH agonist or a salt thereof. obtained thus, and the solution (oil phase) containing the biodegradable polymer regulated at about 25 to about 35 ° C, and the resulting mixture is subjected to the emulsification step to prepare the W / O type emulsion. As the solution (oil phase) containing the biodegradable polymer, a solution is used in which the biodegradable polymer is dissolved in an organic solvent. The organic solvent can be a solvent that has a boiling point of about 120 ° C or less, is hydrophobic, and dissolves the biodegradable polymer. The solvent includes halogenated hydrocarbons (eg, dichloromethane (methylene chloride), chloroform, chloroethane, dichloroethane, trichloroethane, carbon tetrachloride.), Fatty acid esters (eg, ethyl acetate, butyl acetate , etc.), ethers (e.g., ethyl ether, isopropyl ether, etc.), and aromatic hydrocarbons (e.g., benzene, toluene, xylene, etc.). Alternatively, two or more types of these organic solvents can be used when mixing with an appropriate ratio. The organic solvent is preferably methyl chloride. A concentration of a biodegradable polymer in the organic solvent is different depending on the type and molecular weight of the biodegradable polymer, and the type of organic solvent, and usually about 0.01 to about 90% (w / w), preferably about 0.1 to about 80% (w / w), more preferably about 1 to about 70% (w / w), preferably preferably about 35%.

In order to change the compatibility with an aqueous phase, and the distribution of an organic solvent in an external aqueous phase, and the volatilization of an organic solvent, a partially hydrophilic organic solvent such as ethanol, acetonitrile, acetone and acetone can be added to the oil phase. tetrahydrofuran. In addition, in order to dissolve or stabilize a GnRH agonist or a salt thereof in the interior, a surfactant may be added as the fatty acid ester of a sugar. The oil phase thus obtained is generally used after removing the bacteria or powders by means of filtration with a filter. In addition, depending on the stability of the biodegradable polymer, a solution containing a biodegradable polymer can be stored in a sealed container at room temperature or in cold places. A mixing ratio of a solution containing a GnRH agonist or a salt thereof and does not contain a substance that retains the drug, and a solution of a biodegradable polymer is such that the latter is from about 0.1 to about 1000 parts by weight , preferably about 1 to about 100 parts by weight, more preferably about 1 to about 20 parts by weight, particularly preferably about 10 parts by weight per 1 part by weight of the former.

It is different depending on the type of GnRH agonist or a salt thereof, the desired pharmacological effect and the duration or effect, the mixing may develop such that the ratio of a GnRH agonist or a salt thereof to a biodegradable polymer is approximately 0.01. around 50% (w / w), preferably around 0.5 to about 40% (w / w), more preferably around 0.1 to about 30% (w / w), particularly preferably around 10%. An emulsion step is developed by the known dispersion method, for example, an intermittent vibration method, which method uses a mixer such as a propeller-type stirrer and a turbine-type stirrer, colloidal milling method, homogenizer method, and method of ultrasound irradiation. A solution that contains a GnRH agonist or a salt of this and does not contain a substance that. The drug is retained and a solution containing the biodegradable polymer is mixed at a temperature of about 25 to about 35 ° C, preferably about 27 to about 33 ° C. By this temperature adjustment, a controlled release microcapsule having better spherical property and / or needle penetration property can be produced. A preferred aspect of the emulsion step will be described. For example, first, a solution containing a biodegradable polymer is added to a container containing a solution containing a GnRH agonist or a salt thereof and does not contain a substance that retains the drug, and the container is vibrated or rocked , by means of this it develops of coarse emulsion. In the coarse emulsion, it is preferred that the temperature of a mixture of the solution containing a GnRH agonist or a salt thereof and does not contain a substance that retains the drug, and a temperature of the solution containing the biodegradable polymer is adjusted to about 25 to 35 ° C, preferably about 27 to 33 ° C. Since a goal of the coarse emulsion is generally to facilitate an emulsification step (precise emulsion) of the next step, and a time of agitation and vibration and the number of swings is not particularly defined. Therefore, when the precise emulsion is uniformly developed, the coarse emulsion step can be omitted. Then, the mixture after the coarse emulsion is subjected to the emulsification step (precise emulsion) with a stirrer of the propellant type. In the precise emulsion, it is preferred that a temperature of a mixture of the solution containing a GnRH agonist or a salt thereof and does not contain a substance that retains the drug, and a temperature of the solution containing the biodegradable polymer is adjusted to approximately 25 to 35 ° C, preferably around 27 to 33 ° C. By this temperature adjustment, a controlled release microcapsule having better spherical property and / or needle penetration property can be produced. An emulsion time in the precise emulsion can be selected depending on the properties of a GnRH agonist or a salt thereof, and a biodegradable polymer, generally the emulsion is developed with a range of about "0.1 to about 60 minutes. An oil phase which will be mixed in relation to the volume of an internal aqueous phase is from about 1 to about 1000 times, preferably about 2 to about 100 times, more preferably about 3 to about 10 times. The resulting W / O emulsion is generally about 10 to about 10,000 cp, preferably about 100 to about 5,000 cp, particularly about 500 to about 2,000 cp at about 12 to 25 ° C. It is preferred that the emulsion type W / O obtained by the precise emulsion is cooled in a water bath or the like at about 0 to about 18 ° C, and a temperature of The W / O emulsion is adjusted to about 0 to about 30 ° C, preferably about 10 to about 25 ° C, more preferably about 15 to about 20 ° C. Then, the W / O type emulsion thus obtained is dispersed in an aqueous phase (hereinafter, abbreviated as exterior aqueous phase) to produce an emulsion of the W / O / W type, and the emulsion of the W / O type / W is subjected to a water drying to produce a controlled release microcapsule. An emulsifier can be added to the external aqueous phase. As the emulsifier, any emulsifier can be used until it forms a generally stable O / W emulsion, and examples include an anionic surfactant (eg, sodium oleate, sodium stearate, sodium lauryl sulfate, etc.), a non-ionic surfactant. (eg, Tween 80, Tween 60, HCO.-70, etc.), polyvinyl alcohol, polyvinylpyrrolidone, and gelatin. These emulsifiers can be used by mixing two or more types with an appropriate ratio. In the case of the process of the present invention, polyvinyl alcohol is preferably used as an emulsifier. A concentration of an emulsifier in an exterior aqueous phase is, for example, about 0.001 to about 20%, preferably about 0.01 to about 10%, more preferably about 0.05 to about 5%, particularly preferably about 0.1%. . An osmotic pressure adjusting agent can be added to the outer aqueous phase. As an agent that adjusts the osmotic pressure, any agent exhibiting an osmotic pressure when placed in an aqueous solution can be used. Examples of the agent that adjusts the osmotic pressure include polyhydric alcohols, monohydric alcohols, monosaccharides, disaccharides, oligosaccharides, amino acids or derivatives thereof, and sodium chloride. As polyhydric alcohols, for example, trihydric alcohols such as glycerin, pentahydric alcohols such as arabitol, xylitol, adonitol and hexahydric alcohols such as mannitol, sorbitol, and dulcitol can be used. inter alia, hexahydric alcohols are preferred and, in particular, mannitol is suitable. Examples of the monohydric alcohols include methanol, ethanol and isopropyl alcohol and, among these, ethanol is preferred. As the monosaccharides, for example, pentose such as arabinose, xylose, ribose and 2-deoxyribose, and hexose such as glucose, fructose, galactose, maas, sorbose, rhamnose, mucosa are used, among these, earthy is preferred. As oligosaccharide, for example, trisaccharides such as maltotriose and raffinose saccharides, and tetrasaccharides such as stachyose are used and, among these, trisaccharides are preferred. As a derivative of monosaccharides, disaccharides and oligosaccharides, for example glucosamine, galactosamine are used, glucuronic acid and galacturonic acid. Like the amino acids, any amino acid can be used so they are L-isomers, and examples include glycine, leucine and arginine. Among these, L-arginine is preferred. These osmotic pressure regulators can be used alone, or they can be used when mixing them. These osmotic pressure regulators are used at such a concentration that an osmotic pressure of the outer aqueous phase "becomes about 1/50 to about 5 times, preferably 1/25 to about 3 times, more preferably 1/12 to about 2 times an osmotic pressure of a physiological saline solution.Specifically, in the case where the osmotic pressure regulator is a non-ionic substance, a concentration of the osmotic pressure regulator in the outer aqueous phase is approximately 0.01 to around 60% (p / p), preferably about 0.01 to about 40% (w / w), most preferably about 0.05 to about 30% (w / w), preferably about 0.5 to about 1.5% (w / w). In the case where the osmotic pressure regulator is an ionic substance, a concentration is obtained by dividing the concentration mentioned above by the total ionic valence used. A concentration of the osmotic pressure regulator to be added need not be greater than the solubility, and a part of the regulator may be in the dispersed state. By adding the osmotic pressure regulator to the outer aqueous phase, the dispersion of the microcapsule produced can be improved. Its degree is not particularly limited, but for example, it is preferable that about 400 to 700 mg of the microcapsule can be dispersed in 1.5 ml of a dispersing medium for injection in less than 2 minutes. The removal of an organic solvent can be carried out according to the known method. Examples of this method include a method of removing the solvent under a normal temperature or gradually reduced pressure while stirring with a stirrer of the propulsion type or a magnetic stirrer, and a method of removing the solvent using a rotary evaporator while regulating the degree of vacuum and a temperature. The microcapsule that is released controlledly thus obtained is collected by centrifugation, filtration or a wet cyclone, etc., it is washed with distilled water several times to eliminate a free GnRH agonist or a salt thereof, a substance that retains the drug and an emulsifier which adheres to the surface of a microcapsule. Then, the washed microcapsule is dried under reduced pressure, or redispersed in distilled water before it is lyophilized to remove an organic solvent.

During a preparation step in order to prevent the aggregation of the particles, an agent for the prevention of aggregation can be used. As an agent for the prevention of aggregation, for example, water-soluble polysaccharides such as mannitol, lactose, glucose, and starches (eg, corn starch), amino acids such as glycine, and proteins such as fibrin are used. Collagen Among these, el-mannitol is preferred. A quantity of aggregation preventing agents such as mannitol can be added usually is from 0 to about 24% by weight relative to the total microcapsule. It is preferred that the controlled release microcapsule of the present invention contains an excipient. It is desired that the excipient be low in toxicity even when administered to a living body, be easily dried as by lyophilization and, dissolve rapidly when administered to a living body, or dissolve when used. Such an excipient includes, for example, sugar, cellulose derivative, amino acid, protein, polyacrylic acid derivative, organic salt, and inorganic salt. These excipients may be used when mixing two or more types with an appropriate ratio. Herein, examples of the sugar include D-mannitol, sodium alginate, fructose, dextran, dextrin, sucrose, D-sorbitol, lactose, glucose, maltose, starches, and trehalose. Examples of the cellulose derivative include carboxymethylcellulose, hydropropylmethylcellulose, ethylcellulose, hydroxymethylcellulose, hydroxypropylcellulose, cellulose acetate phthalate, hydroxypropylmethylcellulose phthalate, and hydroxymethylcellulose acetate succinate. Examples of amino acids include glycine, alanine, tyrosine, arginine and lysine. Examples of the protein include fibrin, collagen and albumin. Examples of the polyacrylic acid derivative include sodium polyacrylate, and methacrylic acid / acrylic acid copolymer (Eudragit, made by Rohm Pharma, Germany). Examples of the organic salt include citrate, sodium, sodium tartrate, sodium carbonate, and potassium carbonate. Examples of the inorganic salt include sodium chloride, potassium chloride, sodium phosphate, and potassium phosphate. As an excipient, water-soluble polymers are used in which a polymer as a base for a controlled release microcapsule does not dissolve, for example polyvinylpyrrolidone, and polyvinyl alcohol in addition to the aforementioned excipients. The excipient is preferably sugar and, inter alia, D-mannitol is preferred which is freeze-dried easily and has low toxicity. An amount of the excipient used is determined by solubility of the excipient, and toxicity, viscosity, dispersion, and stability of a solution obtained by dissolving the excipient. The excipient is used so that, when a controlled release microcapsule is dried, a content of the excipient in the dry controlled release microcapsule is for example about 0.5 to about 99% (w / w), preferably about 1 to about 90. % (w / w), more preferably around 2 to about 60% (w / w). When D-mannitol is used as the excipient, it is preferred that a content of the excipient in a dried controlled release microcapsule is about 2 to about 40% (w / w), preferably about 15% (w / w). By adding these excipients, the following effects are obtained: 1) the frequency of contact and collision of particles during and after the drying of a controlled release microcapsule is reduced, and by this the uniformity of the particles in lyophilization is retained , 2) it becomes possible to dry a controlled release microcapsule at a temperature of a glass transition point or higher, and by this to remove the water or a complete organic solvent, 3) the stability of the microcapsule is improved. controlled release over time, and by means of this a controlled release microcapsule is obtained which is better in dispersion, and is not limited to storage in cold places, for example, it has a long term use limit at room temperature. The microcapsule (B) containing an excipient can be produced, for example, by mixing an excipient and a microcapsule obtained by the water drying method mentioned above. The microcapsule can be a microcapsule that has been dried under reduced pressure after washing, or it can be a microcapsule that has been redispersed in distilled water after washing, and lyophilized. A mixing method is not particularly limited, for example, mixing is carried out using a mixer. In addition, the microcapsule (B) containing an excipient can also be produced by using an aqueous solution of the excipient in an outer aqueous phase with the production of an emulsion of the W / O / W type used in a water-drying method. The microcapsule (B) containing an excipient is preferably produced by washing a microcapsule obtained by a drying method in water, dispersing the washed microcapsule in distilled water where an excipient has been dissolved or suspended, and the dispersion is subjected to lyophilization or dried under reduced pressure. Alternatively, the washed microcapsule can be dispersed in distilled water, and an excipient can be dissolved or suspended in the resulting dispersion, followed by lyophilization or drying under reduced pressure. inter alia, a uniform mixture is obtained by dispersing the washed microcapsule in distilled water where an excipient has been dissolved, or dissolved in a dispersion obtained by dispersing the washed microcapsule in distilled water, and subjecting it to lyophilization. In addition, the water and an organic solvent can be completely removed in a microcapsule and, at the same time, the controlled release can be improved by heating a microcapsule obtained by the aforementioned water drying method, if desired, at a temperature of one hour. glass transition temperature (Tg) of a polymer used as a base or higher and at this temperature where each particle in the microcapsule does not adhere to each other. In this case, it is preferred that the organic solvent is removed at less than about 1000 ppm, preferably less than about 500 ppm, more preferably less than about 100 ppm. A vitreous transition temperature refers to an intermediate point glass transition temperature obtained when it rises with a heating ratio of 10 to ° C per minute using a differential scanning calorimeter (DSC).

Not limiting the regulation of heating is preferred after the optional addition of an excipient, and after lyophilization or drying under reduced pressure of a microcapsule. For example, heating can be carried out after subdivision. When a heating temperature is lower than a vitreous transition temperature of a polymer used as a base, the removal of water or an organic solvent is insufficient in some cases. On the other hand, when the heating temperature is too high, the risk of melting or deformation of a microcapsule, the degradation and deterioration of a GnRH agonist or a salt thereof is increased. Thus, a heating temperature can not be defined unconditionally , but can be determined inappropriately in view of the physical properties of a polymer used as a base (eg, molecular weight, stability etc.), an average particle diameter and a heating time of a GnRH agonist or a salt of this, and a microcapsule, and a degree of drying, and a method of heating a microcapsule. A heating temperature in a range from a vitreous transition temperature of a polymer used as a base to a temperature of about 40 ° C higher than the vitreous transition temperature, preferably from the glass transition temperature of the polymer at a temperature, is preferred. about 35 ° C higher than the glass transition temperature, more preferably from the vitreous transition temperature of the polymer at a temperature around 25 ° C higher than the glass transition temperature, particularly preferably from the glass transition temperature of the glass transition temperature. polymer at a temperature around 20 ° C higher than the glass transition temperature. A heating time is different depending on a heating temperature and an amount of a microcapsule to be treated and, generally, is about 6 to about 120 hours, more preferably 12 to about 96 hours after a microcapsule has reached by itself a predetermined temperature. In addition, an upper limit of a heating time is not particularly limited to an amount of a remaining organic solvent and the moisture reaches an acceptable or lower value. However, under the condition of a vitreous or greater transition temperature, the microcapsule is softened and deformed by physical contact of the microcapsules or loading to lamination of the microcapsule. It is therefore preferred that the heating be terminated quickly when a remnant of organic solvent and moisture reach an acceptable or lower value. A heating method is not particularly limited, but any method can be used while it can uniformly heat a microcapsule. Preferred examples of the heating method include a method for developing heating and drying - with a lyophilization device or a device of constant temperature and reduced pressure at a low pressure. A particle diameter of the microcapsule (B) is sufficient until it is in a range that satisfies its dispersion and water penetration properties, and is for example 0.1 to about 1000 μm, preferably about 1 to about 300 μm, more preferably about 5 to about 150 μm as expressed by an average diameter. The microcapsule (B) is excellent in the property of dissolution because. a production dissolution rate is high, for example a concentration of methylene chloride remaining in a preparation after finishing a water drying step (eg, after 3 hours) is generally about 2,000 ppm to about 20,000 ppm. In addition, the microcapsule (B) has excellent characteristics of a slow sedimentation rate. The sedimentation rate can be determined, for example, by filling 50 mg of powder of the microcapsule (B) in a bottle, suspending in 5 ml of a dispersion medium, dispersing approximately 40 μl of the resulting suspension in 5 ml of a dispersion medium, and measuring the NTU with a turbidimeter. The microcapsule (B) has a characteristic that, it leaves the turbidity immediately after the suspension which is at 100%, a time until a turbidity of 50% is long. In the microcapsule (C), a ratio of lactic acid / glycolic acid composition is preferably 90/10 to 100/0, particularly preferably 100/0, A weight average molecular weight of a copolymer or a homopolymer is preferably about 7,000 to about 25,000 when a ratio of lactic acid / glycolic acid is 100/0, about 7,000 to about 30,000 when the ratio is 90/10, and about 12,000 to about 30,000 when the ratio is 80/20 A weight average molecular weight in this case can be determined using, for example, the standard substance A described above.A concentration of a GnRH agonist or a salt thereof in a solution with internal aqueous phase is generally about 20%. at 70% (w / w), preferably around 25 to 65% (w / w), more preferably from 35 to 60% (w / w) A concentration of a copolymer or a homopolymer in a solution with fa inner aqueous is generally around 0.5 to 90% (w / w), preferably around 2 to 60% (w / w). A period of duration in which a GnRH agonist or a salt thereof is released in the order of zero is preferably 2 months or more and 4 months or less, more preferably about 3 months. As the long-period controlled release microcapsule (A), specifically, a microcapsule (MC) # 2 produced in Reference Example 2 described below is used. As the short-period controlled release microcapsule (B), specifically, a microcapsule (MC) # 1 produced in Reference Example 1 described below is used. As the short-period controlled release microcapsule (C), specifically, a microcapsule (MC) # 3 produced in Reference Example 3 described below is used. In order to formulate the microcapsule in injectable solution, a controlled release injectable solution is obtained which can currently be used in formulating the microcapsule together with a dispersant (e.g., Tween 80, HCO-60, carboxymethylcellulose, sodium alginate, etc. .), a preservative (eg, methylparaben, propylparaben, etc.), and an isotonic agent (eg, sodium chloride, mannitol, sorbitol, glucose, etc.), in an aqueous suspension, or suspending the microcapsule together with a vegetable oil such as sesame oil and corn oil to obtain an oily suspension. As the controlled release preparation of the present invention, a combination of a preparation obtained by formulating a microcapsule which gradually releases a GnRH agonist or a salt thereof from a long period into a controlled release preparation, in particular, can be used. a preparation such as the controlled release injectable as described above, and a preparation obtained by formulating a microcapsule which gradually releases a GnRH agonist or a salt thereof for a short period, in a controlled release preparation, in particular, a preparation as the controlled-release injectable described above. In addition, a preparation obtained by formulating microcapsules obtained by mixing a microcapsule that gradually releases a GnRH agonist or a salt thereof for a long period and a microcapsule that gradually releases a GnRH agonist or a salt thereof, in a controlled release preparation, in particular a preparation such as the controlled release injectable described above. The preparation containing a GnRH agonist (preferably leuprorelin or a salt thereof), more preferably leuprolein acetate) (preferably, a preparation containing a controlled release microcapsule containing leuprorelin or a salt thereof (preferably leuprorelin acetate) can be administered subcutaneously, directly and easily, intramuscularly or intravascutaneously (preferably subcutaneously and intramuscularly) as an injectable, etc. Similarly, the controlled release preparation of the present invention can be administered directly and easily, intravenously or intramuscularly (preferably subcutaneously and intramuscularly) as an injectable, etc. In the present invention, it is administered when a preparation is obtained by formulating a mixture of a short-time controlled release microcapsule and a long-time controlled release microcapsule in a preparation, or a preparation obtained by mixing a short-time controlled release preparation and a release preparation. Controlled long time in a preparation, this can be administered easily, such as subcutaneously, intramuscularly or intravenously (preferably subcutaneously or intramuscularly) as an injectable, etc. In addition, when a short-time controlled release preparation and a long-time controlled release preparation are administered separately, they can be administered directly and easily subcutaneously, intramuscularly or intravenously (preferably subcutaneously and intramuscularly). Usually, the same route of administration is selected, but the short-time controlled release preparation and the long-time controlled release preparation can occasionally also be administered by separate routes such as subcutaneously and intramuscularly. A dose of the preparation is varied depending on the content and dosage form of a GnRH agonist (preferably leuprorelin or a salt thereof, more preferably leuprorelin acetate), the duration of a GnRH agonist (preferably, leuprorelin or a salt thereof), more preferably leuprorelin acetate), and an animal to be administered [e.g., warm-blooded mammal (e.g., human, mouse, rat, rabbit, sheep, pig, cow, horse, etc.)] , and may be an effective amount as a medicine of the GnRH agonist (preferably, leuprorelin or a salt thereof, more preferably leuprorelin acetate). For example, the daily dose for the warm-blooded mammal can be appropriately selected from a range of 0.01 mg to 100 mg / kg of body weight, preferably around 0.02 mg to 50 mg / kg of body weight, more preferably 0.05 mg to 20 mg. mg / kg of body weight. When the preparation is administered as an injectable, usually, about 0.01 to 50 mg, preferably about 0.1 to 20 mg, more preferably - about 0.1 to 15 mg of a GnRH agonist (preferably, leuprorelin or a salt thereof, more preferably leuprorelin acetate) can be administered subcutaneously or intramuscularly to an adult patient with prostate cancer (weight 60 kg) per day. Further, when administered as an injectable containing controlled release microcapsule containing the GnRH agonist (preferably, leuprorelin or a salt thereof, more preferably leuprorelin acetate), a dose is different depending on the period of controlled release of the drug from the drug. microcapsule controlled release. For example, when administered approximately once a month, usually about 0.01 to 25 mg, preferably about 0.1 to 15 mg, more preferably about 0.1 to 10 mg of a GnRH agonist (preferably, leuprorelin or a salt thereof), more preferably leuprorelin acetate) can be administered subcutaneously or intramuscularly to an adult patient with prostate cancer (weight 60 kg) for one time. For example, when administered approximately once every 3 months, usually around 0.1 to 75 mg, preferably about 0.1 to 45 mg, more preferably about 1 to 30 mg of a GnRH agonist (preferably, leuprorelin or a salt thereof, more preferably leuprorelin acetate) can be administered subcutaneously or intramuscularly to an adult patient with prostate cancer (weight 60 kg) for once. For example, when administered about once every 6 months, usually about 0.2 to 150 mg, preferably about 0.2 to 90 mg, more preferably about 2 to 60 mg of a GnRH agonist (preferably, leuprorelin or a salt thereof). , more preferably leuprorelin acetate) can be administered subcutaneously or intramuscularly to an adult patient with prostate cancer (weight 60 kg) for one time. Also in the case of other animals, an amount obtained by converting into a 60 kg by weight amount may be administered, and may be administered by appropriately increasing or decreasing the aforementioned dose depending on the term of continuous release.

The preparation comprising a combination of a short-period controlled release microcapsule and the long-period controlled release microcapsule of the present invention can be administered by converting the aforementioned dose of a GnRH agonist from a period of controlled release of the microcapsule from controlled release of long period, and appropriately distribute in the short-period controlled release microcapsule and the long-period controlled release microcapsule. When the amino acids, peptides, and protecting groups in the polypeptides described herein are expressed by abbreviations, this is based on the abbreviations of the IUPAC-IUB Commission on Biochemical Nomenclature or abbreviations conventional in the art. In addition, when the amino acid can have an optical isomer, an L-isomer is denoted unless otherwise indicated. Examples of abbreviations are shown below: Abu: Aminobutyric acid Aibu: 2-aminobutyric acid Wing: Alanine Arg: Arginine Gly: Glycine His: Histidine lie: Isoleucine Leu: Leucine Met: Methionine Nle: Norleucine Nval: Norvaline Phe: Phenylalanine Phg: Phenylglycine Pro: Proline (Pyr) Glu: Pyroglutamic acid Ser: Thrine: Threonine Trp: Tryptophan Tyr: Tyrosine Val: Valine D2Nal: Residue of D-3- (2 -nafyl) alanine DSer (tbu) O-tert-butyl- D-serine DHis (ImBzl) Ni? N-benzyl-D-histidine PAM: Phenylacetamidomethyl Boc: t-Butyloxycarbonyl Fmoc: 9-Fluorenylmethyloxycarbonyl Cl-Z: 2-Chloro-benzyloxycarbonyl ~ Br-Z 2-Bromo-benzyloxycarbonyl Bzl: Benzyl Cl2Bzl 2, 6-Dichlorobenzyl Cough: p-toluenesulfonyl HO? B:? -hydroxy-5-norbornene-2,3-dicarboximide HOBt: 1-hydroxybenzotriazole HOOBt: 3-hydroxy-3,4-dihydrogen-4-oxo-l , 2, 3-benzotriazine MEBzl: 4-Methylbenzyl Bom: Benzyloxymethyl Bum: t-butoxymethyl Trt: Trifyl DNP: Dinitrophenyl DCC: N, N '-dicyclohexylcarbodiimide EXAMPLES The following Examples that include Experimental Examples will illustrate the present invention more specifically.

Reference Example 1 Production of microcapsule (B) Weighed 119.1 g of 5-oxo-Pro-His-Trp-Ser-Tyr-DLeu-Leu-Arg-Pro-NH-C2H5 (hereinafter, abbreviated as peptide A) Acetate in a flask of the aubergine type, and 120 g of water for injection were added to completely dissolve it. To this was added 975 g of lactic acid-glycolic acid copolymer (ratio of lactic acid composition "glycolic acid = 75:25, Mw = approximately 10,400, Mn = approximately 4,100, Mw / Mn = 2.5 (value measured by the method of GPC in Reference Example 5 (value measured using standard substance C)) dissolved in 1600 g of dichloromethane, and this was stirred and emulsified with an Automimixer at approximately 5800 rpm for 10 minutes to obtain a W / O emulsion. emulsion w / O was cooled to about 19 ° C, was emptied in 200 L of an aqueous solution of polyvinyl alcohol 0.1% (w / w) (EG-40, manufactured by Nippon Synthetic Chemical Industry Co., Ltd.) which has been regulated at about 19 ° C beforehand, and stirred and emulsified at about 7000 rpm using HOMOMIC LINE FLOW (manufactured by Tokushu Kika Kogyo Co., Ltd.) to obtain a W / O / W emulsion. / O / W was stirred at room temperature around 2500 rpm for 3 hours and the dichloromethane was volatilized or diffused into an external aqueous phase to solidify an oil phase. After it was passed through a screen having 75 μm openings, a microcapsule was continuously seated with a centrifuge at about 2000 rpm and collected. The collected microcapsule was dispersed in a small amount of distilled water, and passed through a sieve having 90 μm openings, and 174.5 g of mannitol was added to dissolve it. This was lyophilized to obtain a microcapsule powder (hereinafter, MC # 1). A content of peptide A was 8.5%.

Reference Example 2 Production of the microcapsule (A) 123.3 g of the acetate of peptide A were weighed in a flask of the aubergine type, and 129.4 g of an aqueous solution of acetic acid were added to completely dissolve it. To this were added 1080 g of DL-lactic acid polymer (Mw = approximately 21,400 (value measured using standard substance B)) dissolved in 1890 g of dichloromethane, and crudely dispersed in 1890 g of dichloromethane, coarsely dispersed for about 2 minutes, and stirred and emulsified with an Automimixer at approximately 5800 rpm for 4 minutes to obtain a W / O emulsion. This W / O emulsion was cooled to about 18 ° C, was emptied in 200 L of an aqueous solution of polyvinyl alcohol 0.1% by weight (EG-40, manufactured by the Nippon Synthetic Chemical Industry Co., Ltd.) which was has been regulated at about 19 ° C in advance, and stirred and emulsified at about 7000 rp'm using HOMOMIC LI? E FLOW (manufactured by Tokushu Kika Kogyo Co., Ltd.) to obtain a W / O / W emulsion. This W / O / W emulsion was stirred at room temperature around 2500 rpm for 3 hours, the dichloromethane was volatilized or diffused into an external aqueous phase to solidify an oil phase. After it was passed through a sieve having 75 μm openings, a microcapsule was continuously seated with a centrifuge at about 2000 rpm and this was collected. The collected microcapsule was dispersed in a small amount of distilled water, and passed through a sieve having 90 μm openings, and 169.7 g of mannitol was added to dissolve it. This was lyophilized to obtain a powder of microcapsules (hereinafter, MC # 2). A content of the peptide was 7.5%.

Reference Example 3 Production of the microcapsule (C) 86.7 g of the acetate of peptide A were weighed in a Kolben of the eggplant type, and 100 g of injection water was added to completely dissolve it. To this were added 765 g of DL-lactic acid polymer (Mw = approximately 14,200 (value measured using standard substance A)) dissolved in 1280 g of dichloromethane, and stirred and emulsified with an Automimixer at approximately 5800 rpm for 13.5 minutes. to obtain a W / O emulsion. This W / O emulsion was cooled to about 15 ° C, emptied in 200 L of an aqueous solution of polyvinyl alcohol 0.1% (w / w) (EG-40, manufactured by the Nippon Synthetic Chemical Industry Co., Ltd. ) which has been regulated at about 15 ° C in advance, and stirred and emulsified at about 7000 rpm using HOMOMIC LINE FLOW (manufactured by Tokushu Kika Kogyo Co., Ltd.) to obtain a W / O / W emulsion. This W / O / W emulsion was stirred at room temperature around 2500 rpm for 3 hours, the dichloromethane was volatilized or .difundió in an external aqueous phase to solidify an oil phase. After it was passed through a sieve having 75 μm openings, a microcapsule was continuously seated with a centrifuge at about 2000 rpm and this was collected. The collected microcapsule was dispersed in a small amount of distilled water, and passed through a sieve having 90 μm openings, and 130 g of mannitol was added to dissolve it. This was lyophilized under secondary drying conditions at 50 ° C for 48 hours to obtain a microcapsule powder (hereinafter, MC # 3). A content of the peptide was 7.8%.

Reference Example 4 Production of the microcapsule (C) 14.5 g of the acetate of peptide A were weighed in a Kolben of the eggplant type, and 15.9 g of injection water were added to completely dissolve it. To this were added 123 g of DL-lactic acid polymer (Mw = approximately 1400) dissolved in 204 g of dichloromethane. it was crudely emulsified for 1 minute, and stirred and emulsified with an Automimixer at approximately 10,000 rpm for 3 minutes to obtain a W / O emulsion. This W / O emulsion was cooled to about 16 ° C, emptied into 25 L of an aqueous solution of polyvinyl alcohol 0.1% (w / w) (EG-40, manufactured by the Nippon Synthetic Chemical Industry Co., Ltd. ) which has been adjusted to about 16 ° C in advance, and stirred and emulsified at about 7000 rpm using HOMOMIC LI? E FLOW (manufactured by Tokushu Kika Kogyo Co., Ltd.) to obtain a W / O / W emulsion. This W / O / W emulsion was stirred at room temperature around 2000 rpm for 3 hours and the dichloromethane was volatilized or diffused into an external aqueous phase to solidify an oil phase. After it was passed through a sieve having 75 μm openings, a microcapsule was continuously seated with a centrifuge at about 2000 rpm and this was collected. The collected microcapsule was dispersed in a small amount of distilled water, and passed through a sieve having 90 μm openings, and 17.5 g of mannitol was added to dissolve it. This was lyophilized under secondary drying conditions at 50 ° C for a secondary drying time of 0 hours, 20 hours, 22 hours, 24 hours, 26 hours and 48 hours, to collect each microcapsule powder separately. Reference Example 5 Measurement of weight average molecular weight (Mw) of the polymer (GPC method) Approximately 0.05 g of this product was weighed, tetrahydrofuran (THF) was added to dissolve it with 5 ml, and the sample solution was obtained. Separately, each of the standard 0.1 g polystyrene products (F-10, F-2, A-5000 and A-1000) having the known molecular weights was weighed, THF was added to dissolve it to 40 ml, and A standard solution A was obtained. In addition, each of the standard polystyrene products of about 0.1 g (F-4, Fl, A-2500 and A-500) having the known molecular weights, THF was added to dissolve it to 40 ml, and a standard solution B was obtained. 100 μl of each sample solution and the standard solutions A and B were tested by means of the gel permeation chromatography method under the following conditions. A molecular weight calibration curve was produced with a molecular weight of each of the standard polystyrene products and their retention time. Then, a high peak (Hi) of an eluted compound obtained from the sample solution was measured, and its molecular weight (Mi) was obtained from its retention time and the calibration curve. A weight average molecular weight (Mw) of the present product was obtained with the following equation. [Calculation equation] 'Mw =? (HixMi) /? Hi [Test condition] Detector: differential refractometer (which has equivalent performance to the HLC-8120GPC system) Column: GuardColumn TSK HHR-L (40x6.0 mm di) Gel G4000HHR (300x7.8 mm di), Gel G3000HHR (300X7.8 mm di) Gel G2000HHR (300X7.8 mm di), and Gel GIOOOHHR (300X7.8 mm di) are connected in a series to reduce a pore diameter of a filling (or those with the lowest performance can be used). Column temperature: a constant temperature around 50 ° C Mobile phase: THF Flow rate: 1.0 ml / min [System Convenience] (1) System Development: When operating the system in 100 μl of standard solution A under the conditions mentioned above, a degree of separation between a peak of F-10 and a peak of F-2 is 2.0 or greater, and a theoretical step number, and a symmetric coefficient of both peaks are 800 steps or more and 1.5 or less, respectively. (2) Reproducibility of the test: When the test was repeated twice in lOOμl of the standard solution A under the conditions mentioned above, a relative standard deviation between the retention times of each peak is less than 3.3% or less.

[Method of operation]. Standard solution: the solution is stable at room temperature (approximately 25 ° C) within at least 24 hours after preparation. In addition, the solution is stable for at least 7 months in a refrigerator (approximately -18 ° C) after preparation.

Solution Sample: The solution is stable at room temperature (around 25 ° C) at least within 24 after the preparation. The measurement range of the area: 48 minutes (injection interval is 50 minutes) Molecular weight calibration curve: it was produced by a polygonal line. In addition to the weight average molecular weight (Mw), an average number-average molecular weight was also measured [Mn =? Hi /? (Hi / Mi)] .- [Order of injection] (1) A test was repeated twice in a standard solution A, and it was confirmed that the first test was adopted to the system development rule. A retention time of each peak was obtained, and it was confirmed that this was adapted to the reproducibility rule of the test (a relative standard deviation between the retention time of the respective peaks is 3.3% or less). (2) A standard solution was injected, and a retention time of each peak was obtained. (3) A mobile phase was injected, a remnant of all the peaks of the standard solution B injected into (2), and it was confirmed that a peak area value satisfies the specification (10% or less). (4) Measurement of the sample solution (maximum 12) (5) a mobile phase was injected, a remnant of the sample solution finally injected into (4) was verified, and it was confirmed that a value of the peak area is adapted to the rule (10% or less). (6) The standard solutions A and B are injected, and a retention time of each molecular weight is obtained. (7) A calibration curve of the molecular weight of the retention times of the standard solution A finally injected in (1) is produced, the standard solution B in (2) is injected, and the standard solutions A and B are injected in (6), and a weight average molecular weight (Mw) of the sample is calculated, provided it is confirmed that a relative deviation of the retention time [RD:% of a difference (absolute value) of an average value of any retention time relative to the average value] of the standard solution A injected finally into (1) and the standard solution A injected into (6) is 3.3% or less. When it is not adapted, all the data between the system checks are invalidated, and the test is developed again (1) (provided that it is not necessary to investigate the performance of the system).

[Reagent • test solution] Polystyrene standard products: TSK standard polystyrene / manufactured by Tosoh Corporation As standard polystyrene products, the evaluated values are used by means of the GPC method. Type Mw F-10 98900 F-4 37200 F-2 17100 F-l 9490 A-5000 5870 A-2500 2500 A-1000 1051 A-500 495 Tetrahydrofuran: for liquid chromatography, developed by Wako Puré Chemical Industries, Ltd.

EXAMPLE 1 To 0.141 g of MC # 1 (8.5% peptide A acetate content produced in Reference Example 1) was added 1,920 g of MC # 2 (7.5% peptide A acetate) produced in Reference Example 2 and the materials were mixed to prepare two types of mixed powders of microcapsules (hereinafter, combo A). After this, a combination ratio was 1 to 12 (as weight ratio of acetate of peptide A).

Example 2 To 0.184 g of MC # 3 (acetate of peptide A content 7.8%) produced in Reference Example 3, 1.20 g of MC # 2 (acetate of peptide A content 7.5%) produced in Reference Example 2 was added. they were mixed to prepare two types of mixed powders of microcapsules (hereinafter, combo B). After this, a combination ratio was from 1 to 9 (as weight ratio of acetate of peptide A).

Example 3 To 0.154 g of MC # 3 (acetate of peptide A content 7.8%) produced in Reference Example 3, was added - 1,920 g of MC # 2 (acetate of peptide A content 7.5%) produced in the Reference Example 2 and mixed to prepare two types of microcapsule blended powders (hereafter, combo C). After this, a combination ratio was 1 to 12 (as weight ratio of acetate of peptide A).

Example 4 r To 0.141 g of MC # 1 (peptide A acetate - 8.5% content) produced in Reference Example 1, 2,560 g of MC # 2 (peptide A acetate content 7.5%) produced in Example 1 was added. Reference 2 was mixed to prepare two types of mixed powders of microcapsules (hereinafter, combo D). After this, a combination ratio was 1 to 16 (as weight ratio of acetate of peptide A).

EXPERIMENTAL EXAMPLE 1 120 mg of combo B, the two mixed types of microcapsule powders produced in Example 2 (9 mg as peptide A acetate) were suspended in about 0.3 ml of a dispersing medium, the suspension was injected subcutaneously into a rat , a concentration of peptide A in the serum was measured. About 0.3 ml of a 120 mg MC # 2 dispersion medium (9 mg as peptide A acetate) was suspended, and the similar test was developed in another rat. The change in blood concentration of combo B was compared with that of type alone. The change in blood concentration up to 5 weeks after administration is shown in Fig. 1. When "blood concentrations were compared within week 1 after administration, combo B was higher than the MC # 2, both had different rates of controlled release, and the effect on the controlled release of the 2 mixed types of microcapsules was confirmed.

EXPERIMENTAL EXAMPLE 2 129 mg of combo A, the two mixed types of microcapsule powders produced in Example 1 (9.75 mg as peptide A acetate) were suspended in suspension of 0.3 ml of a dispersing medium, the suspension was injected subcutaneously into a rat. , a concentration of peptide A in the serum was measured. They were suspended in approximately 0.3 ml of a 120 mg MC # 2 dispersion medium (9 mg as peptide A acetate), the similar test was developed in another rat. The change in blood concentration of combo A was compared with that of type alone. The change in blood concentration up to 6 weeks after administration is shown in Fig. 2. When blood concentrations were compared within week 3 after administration, combo A was higher than MC # 2. , both had different rates of controlled release, and the effect on the controlled release of the 2 mixed types of microcapsules was confirmed.

EXPERIMENTAL EXAMPLE 3 169 mg of combo D, the two mixed types of microcapsule powders produced in Example 4 (12.75 mg as peptide A acetate) were suspended in approximately 0.3 ml of a dispersing medium., the suspension was injected subcutaneously into a rat, a concentration of peptide A in the serum was measured. They were suspended in approximately 0.3 ml of a 120 mg MC # 2 dispersion medium (9 mg as peptide A acetate), the similar test was developed in another rat. The change in blood concentration of combo D was compared with that of type alone. The change in blood concentration up to 6 weeks after administration is shown in Fig. 3. When the blood concentrations were compared within week 3 after administration, combo D was higher than MC # 2. , both had different rates of controlled release, and the effect on the controlled release of the 2 mixed types of microcapsules was confirmed.

Industrial application By combining the microcapsules that gradually release a GnRH agonist or a salt thereof and have different controlled release periods, an excellent preparation in controlled release can be obtained which has an increasing amount of drug that is released in one stage, early of administration, and releases a constant amount of drug over a long period. It is noted that in relation to this date, the best method known to the applicant to carry out the aforementioned invention, is that which is clear from the present description of the invention.

Claims (14)

2. Having described the invention as above, the content of the following claims is claimed as property. 1. Controlled release preparation, characterized in that it comprises a combination of a microcapsule that grlly releases a GnRH agonist or a salt thereof for a long period, and a microcapsule that grlly releases a GnRH agonist or a salt thereof for a short period. 2. Preparation according to claim 1, characterized in that the GnRH agonist or a salt thereof is a peptide represented by the formula: 5-oxo-Pro-His-Trp-Ser-Tyr-Y-Leu-Arg-Pro- Z [wherein Y represents a selected residue of DLeu, DAla, DTrp, DSer (tBu), D2Nal and DHis (ImBzl), and Z represents NH-C2H5 or Gly-NH2] or a salt thereof;
3. 3. Preparation according to claim 1, characterized in that the GnRH agonist or a salt thereof is an acetate of a peptide of the formula: 5-oxo-Pro-His-Trp-Ser-Tyr-Dlue-Leu-Arg- Pro-NH-C2H5
4. 4. Preparation according to claim 1, characterized in that the long period is 5 months or longer, and the short period is less than 5 months.
5. 5. Preparation according to claim 1, characterized in that the long period is 5 months or longer, and 8 months or less, and the short period is 1 week or more and less than 5 months.
6. 6. Preparation according to claim 1, characterized in that the microcapsule is a microcapsule containing a polymer of lactic acid or a polymer of lactic acid-glycolic acid as a base.
7. 7. Preparation according to claim 1, characterized in that a combination ratio of the microcapsule that grlly releases a GnRH agonist or a salt thereof for a short period to the microcapsule that grlly releases a GnRH agonist or a salt thereof during a period of time. long period is from 1: about 5 to: about 20 expressed as a ratio of a weight of the GnRH agonist or a salt thereof contained in each microcapsule.
8. 8. Preparation according to claim 1, characterized in that: the microcapsule that grlly releases the GnRH agonist or a salt thereof for a long period is: a microcapsule containing (i) a GnRH agonist or a salt thereof, and ( ii) a lactic acid polymer having a weight average molecular weight of about 18,000 to about 30,000; and the microcapsule that grlly releases a GnRH agonist or a salt thereof for a short period is: (1) a microcapsule containing (i) a GnRH agonist or a salt thereof, and (ii) a lactic acid-acid polymer glycolic (75/25) (% mol)) having a weight average molecular weight of about 8,000 to about 12,000, or (2) a microcapsule containing - (i) a GnRH agonist or a salt thereof, and ( ii) a lactic acid polymer having a weight average molecular weight of about 13,000 to about 18,000;
9. 9. Preparation according to claim 1, characterized in that: the microcapsule that grlly releases a GnRH agonist or a salt thereof for a long period is: a microcapsule containing (i) a GnRH agonist or a salt thereof, and ( ii) a lactic acid polymer having a weight average molecular weight of about 15,000 to about 50000 wherein a content of a polymer having a weight average molecular weight of 5000 or less is about 5% or less by weight; and the microcapsule that grlly releases a GnRH agonist or a salt thereof for a short period is: (1) a microcapsule containing (i) a GnRH agonist or a salt thereof, and (ii) a lactic acid-acid polymer glycolic wherein a weight average molecular weight (Mw) is from about 8,000 to about 11,500, and a ratio of a weight average molecular weight (Mw) to a number average molecular weight (Mn) is greater than 1.9, and a molar ratio of the composition of lactic acid to glycolic acid is 99.9 / 0.1 to 60/40, and which does not contain a substance that retains the drug, or (2) a microcapsule that has an order of zero release of a GnRH agonist or a salt thereof for 2 months, and which is prepared by microencapsulating a W-emulsion. / Or prepared from a solution with internal aqueous phase containing a GnRH agonist. or a salt thereof in about 20 to 70% by weight, and an oil phase solution containing, as a substance controlling the release, a copolymer or a homopolymer wherein a ratio of lactic acid / glycolic acid is 80 / 20 to 100/0, and a weight average molecular weight of from about 7,000 to about 30,000.
10. 10. Controlled release preparation according to any of claims 1 to 9, characterized in that it gradually releases a GnRH agonist or a salt thereof for a long period.
11. 11. Controlled release preparation according to claim 10, characterized in that the long period is 5 months or more.
12. 12. Agent for preventing or treating prostate cancer, prostatomegaly, endometriosis, hysteromyoma, metrofibroma, precocious puberty, dysmenorrhea or breast cancer, or a contraceptive agent, characterized in that it comprises a controlled release preparation according to claim 1.
13. 13. Process for producing the controlled release preparation according to claim 1, characterized in that it comprises mixing a microcapsule that gradually releases a GnRH agonist or a salt thereof for a long period and a microcapsule which gradually releases a GnRH agonist or a salt of this for a short period.
14. 14. Use of a controlled release preparation according to claim 1 to produce an agent for preventing or treating prostate cancer, prostatomegaly, endometriosis, hysteromyoma, metrofibroma, precocious puberty, dysmenorrhea or breast cancer, or contraceptive agent.