JPWO2006075690A1 - Stable pharmaceutical composition - Google Patents

Abstract

[Summary] [Problem] To provide a stable pharmaceutical composition containing an EP4 agonist. [Solution] Methyl 4-{[2-((1R, 2R, 3R) -3-hydroxy-2-{(1E, 3S) -3-hydroxy-4- [3- (methoxy) having EP4 activity] Methyl) phenyl] but-1-enyl} -5-oxocyclopentyl) ethyl] sulfanyl} butanoate in an aqueous solution containing a cyclodextrin inclusion compound is mixed with a saccharide and, optionally, a pH adjuster, to a pH of about 3 to After adjusting to about 6.5, freeze-drying is performed. The resulting lyophilized product is stable during storage and does not produce degradation products, so it can be used in the clinical setting as a higher quality injectable preparation. [Selection figure] None

Description

  The present invention relates to (1) methyl 4-{[2-((1R, 2R, 3R) -3-hydroxy-2-{(1E, 3S) -3-hydroxy-4- [3- (methoxymethyl) phenyl]. ] But-1-enyl} -5-oxocyclopentyl) ethyl] sulfanyl} butanoate, a solvate thereof, or an inclusion compound thereof, and (2) a saccharide, and optionally (3) a pH regulator. The present invention relates to a stable pharmaceutical composition, a method for producing the same, and a use thereof.

  Methyl 4-{[2-((1R, 2R, 3R) -3-hydroxy-2-{(1E, 3S) -3-hydroxy-4- [3- (methoxymethyl) phenyl] but-1-enyl} -5-oxocyclopentyl) ethyl] sulfanyl} butanoate is a compound known as a 5-thia-ω-substituted phenyl-prostaglandin E derivative having EP4 activity, which is a preservative, wetting agent, emulsifier, dispersion Injectable for parenteral administration, various diseases, by dissolving, suspending or emulsifying in a sterile aqueous or non-aqueous solvent together with adjuvants such as agents, stabilizers, and solubilizers For example, it is described that it is used as a preventive and / or therapeutic agent for bone loss diseases (International Publication No. 00/03980, International Publication No. 01/37877). Issue pamphlet). In addition, these publications include methyl 4-{[2-((1R, 2R, 3R) -3-hydroxy-2-{(1E, 3S) -3-hydroxy-4- [3- (methoxymethyl) in advance. Sterile solid compositions containing) phenyl] but-1-enyl} -5-oxocyclopentyl) ethyl] sulfanyl} butanoate are prepared and dissolved in sterilized or sterile distilled water for injection or other solvents at the time of use. It is described that it may be used.

On the other hand, it is generally known that a compound having a prostaglandin E (PGE) -like structure is unstable to heat or the like and produces a decomposition product having a prostaglandin A (PGA) -like structure. . Therefore, various techniques are used in pharmaceutical preparations containing a compound having a PGE-like structure (for example, PGE ₁ etc.) in order to prevent the formation of PGA-like degradation products. For example, JP-A-49-66816 discloses a method in which vitamin C or citric acid is added to an aqueous solution of a cyclodextrin clathrate compound of PGE or a cyclodextrin clathrate compound of a PGE analog and freeze-dried (JP-A No. 49-66816). JP-A-49-0666816), JP-A-54-103844 discloses a method in which oligosaccharide is added to cyclodextrin of PG or a PG-like compound or citric acid solution and freeze-dried (JP-A-54-86). No. 103844). Further, for example, JP-A-57-156460 discloses a stabilizing composition comprising PG or a PG-like compound, cyclodextrin, and oligosaccharide (see JP-A-57-156460). -240835 discloses a PGE1-containing injection composition (see JP-A-11-240835) in which PGE1 is dissolved in a sodium citrate / citrate buffer, sterile filtered and freeze-dried.

  Methyl 4-{[2-((1R, 2R, 3R) -3-hydroxy-2-{(1E, 3S) -3-hydroxy-4- [3- (methoxymethyl) phenyl] but-1-enyl} Although -5-oxocyclopentyl) ethyl] sulfanyl} butanoate has a PGE-like structure in part, a report on its chemical stability, including the presence or absence of degradation products having a PGA-like structure, There was nothing, and the degradation product was not known at all.

We have methyl 4-{[2-((1R, 2R, 3R) -3-hydroxy-2-{(1E, 3S) -3-hydroxy-4- [3- (methoxymethyl) phenyl]. As a result of studies to produce an injectable preparation containing but-1-enyl} -5-oxocyclopentyl) ethyl] sulfanyl} butanoate as an active ingredient, methyl 4-{[2-((1R, 2R, 3R) Α-cyclo of -3-hydroxy-2-{(1E, 3S) -3-hydroxy-4- [3- (methoxymethyl) phenyl] but-1-enyl} -5-oxocyclopentyl) ethyl] sulfanyl} butanoate denoted by the aqueous solution and it containing the dextrin clathrate compound for lyophilization in a freeze-dried product produced during the storage period, as in the case of the PGE _1, having a PGA-like structure It found that decomposition products are generated, further in addition thereto, have found that a novel degradation products of sulfoxide and the like is produced.

  In general, storage stability of pharmaceutical products is important from the viewpoint of safety and effectiveness, and it is said that degradation of the main active substance during the storage period and generation of decomposition products accompanying it should be suppressed as much as possible. . This means that methyl 4-{[2-((1R, 2R, 3R) -3-hydroxy-2-{(1E, 3S) -3-hydroxy-4- [3- (methoxymethyl) phenyl] buta- The same applies to an injectable preparation comprising 1-enyl} -5-oxocyclopentyl) ethyl] sulfanyl} butanoate as an active ingredient. In order to use this as a pharmaceutical product, the above-mentioned decomposition product must be produced during the storage period. It was necessary to suppress as much as possible.

International Publication No. 00/03980 Pamphlet International Publication No. 01/37877 Pamphlet JP 49-0666816 A JP 54-103844 A JP 57-156460 A Japanese Patent Laid-Open No. 11-240835

  The subject of the present invention is methyl 4-{[2-((1R, 2R, 3R) -3-hydroxy-2-{(1E, 3S) -3-hydroxy-4- [3- (methoxymethyl) phenyl]]. Buta-1-enyl} -5-oxocyclopentyl) ethyl] sulfanyl} butanoate is used as an active ingredient to produce degradation products during storage, that is, production of degradation products such as degradation products having a PGA-like structure and sulfoxide compounds. An object of the present invention is to provide a high-quality pharmaceutical composition, particularly an injectable preparation, which is stable in any aspect of reduction, purity and form.

  As a result of intensive studies to solve the above problems, the present inventors have found that methyl 4-{[2-((1R, 2R, 3R) -3-hydroxy-2-{(1E, 3S)- An aqueous solution containing an α-cyclodextrin inclusion compound of 3-hydroxy-4- [3- (methoxymethyl) phenyl] but-1-enyl} -5-oxocyclopentyl) ethyl] sulfanyl} butanoate and a saccharide; As a lyophilized product that suppresses the formation of degradation products during the storage period by adding a pH adjusting agent as desired, adjusting the pH to a range of about 3 to about 6.5, and subjecting the aqueous solution to lyophilization. I found out that it can be provided. In addition, it has been found that by reducing the water content in the lyophilized product, a stable composition in which the generation of decomposition products can be further suppressed can be provided, and the present invention can be further improved by further study based on these findings. completed.

That is, the present invention
[1] (1) Methyl 4-{[2-((1R, 2R, 3R) -3-hydroxy-2-{(1E, 3S) -3-hydroxy-4- [3- (methoxymethyl) phenyl] Butane-1-enyl} -5-oxocyclopentyl) ethyl] sulfanyl} butanoate, its solvate, or their inclusion compounds, and (2) a saccharide, and optionally (3) a pH regulator A stable pharmaceutical composition comprising:
[2] The composition according to the above [1], which is a freeze-dried product;
[3] The composition according to the above [1], wherein the inclusion compound is an α-cyclodextrin inclusion compound;
[4] The composition according to the above [1], wherein the saccharide is lactose or maltose;
[5] The composition according to [4], wherein the saccharide is maltose having a pH of 4.5 to 6.5;
[6] Methyl 4-{[2-((1R, 2R, 3R) -3-hydroxy-2-{(1E, 3S) -3-hydroxy-4- [3- (methoxymethyl) phenyl] buta-1 -Enyl} -5-oxocyclopentyl) ethyl] sulfanyl} butanoate The composition according to [1] above, comprising 500 parts by weight to 50,000 parts by weight of saccharides;
[7] The composition according to the above [1], wherein the pH regulator is one or more selected from organic acids, metal hydroxides and organic acid metal salts;
[8] The composition according to [7] above, wherein the pH regulator is (1) citric acid and sodium hydroxide, or (2) citric acid and sodium citrate;
[9] (1) Methyl 4-{[2-((1R, 2R, 3R) -3-hydroxy-2-{(1E, 3S) -3-hydroxy-4- [3- (methoxymethyl) phenyl] An α-cyclodextrin inclusion compound of but-1-enyl} -5-oxocyclopentyl) ethyl] sulfanyl} butanoate, and (2) maltose at pH 4.5 to 6.5, and (3) citric acid and A stable pharmaceutical composition comprising sodium hydroxide;
[10] The composition according to [9] above, which is a freeze-dried product;
[11] Methyl 4-{[2-((1R, 2R, 3R) -3-hydroxy-2-{(1E, 3S) -3-hydroxy-4- [3- (methoxymethyl) phenyl] buta-1 -Ethyl} -5-oxocyclopentyl) ethyl] sulfanyl} butanoate The composition according to [9] above, containing 1000 to 10,000 parts by weight of maltose having a pH of 4.5 to 6.5;
[12] Methyl 4-{[2-((1R, 2R, 3R) -3-hydroxy-2-{(1E, 3S) -3-hydroxy-4- [3- (methoxymethyl) phenyl] buta-1 -Enyl} -5-oxocyclopentyl) ethyl] sulfanyl} butanoate The composition according to [11] above, containing 5000 parts by weight of maltose having a pH of 4.5 to 6.5;
[13] The composition according to the above [2] or [10], which is substantially free of water or has a water content of 1% or less;
[14] After storage at 60 ° C for 2 weeks, methyl 4-{[2-((1R, 2R, 3R) -3-hydroxy-2-{(1E, 3S) -3-hydroxy-4- [3- The stable composition according to [2] or [10], wherein the residual ratio of (methoxymethyl) phenyl] but-1-enyl} -5-oxocyclopentyl) ethyl] sulfanyl} butanoate is 85% or more;
[15] The stable composition according to [14], wherein the residual ratio is 95% or more;
[16] The stable composition according to the above [2] or [10], which has a form substantially the same as the initial state after storage at 60 ° C. for 2 weeks;
[17] The composition according to [1], [2], [9] or [10], which is an injectable preparation;
[18] The composition according to [1], [2], [9] or [10] above, which is a preventive, therapeutic and / or progression inhibitor for EP4-mediated diseases;
[19] The composition of the above-mentioned [18], wherein the EP4-mediated disease is a bone disease;
[20] The composition of the above-mentioned [19], wherein the bone disease is a vertebral fracture.
[21] An injection container filled with the composition according to [2] or [10];
[22] The container according to [21], which is a vial;
[23] 1 to 12 μg methyl 4-{[2-((1R, 2R, 3R) -3-hydroxy-2-{(1E, 3S) -3-hydroxy-4- [3- (Methoxymethyl) phenyl] but-1-enyl} -5-oxocyclopentyl) ethyl] sulfanyl} butanoate, the container according to [21] above;
[24] 2 μg, 5 μg, 6 μg or 10 μg of methyl 4-{[2-((1R, 2R, 3R) -3-hydroxy-2-{(1E, 3S) -3-hydroxy-4 per unit form] -[3- (methoxymethyl) phenyl] but-1-enyl} -5-oxocyclopentyl) ethyl] sulfanyl} butanoate;
[25] (1) Methyl 4-{[2-((1R, 2R, 3R) -3-hydroxy-2-{(1E, 3S) -3-hydroxy-4- [3- (methoxymethyl) phenyl] Butane-1-enyl} -5-oxocyclopentyl) ethyl] sulfanyl} butanoate, its solvate, or their inclusion compounds, and (2) a saccharide, and optionally (3) a pH regulator A method for producing a stable freeze-dried product, wherein the aqueous solution having a pH of 3 to 6.5 is subjected to freeze-drying;
[26] (1) Methyl 4-{[2-((1R, 2R, 3R) -3-hydroxy-2-{(1E, 3S) -3-hydroxy-4- [3- (methoxymethyl) phenyl] Butane-1-enyl} -5-oxocyclopentyl) ethyl] sulfanyl} butanoate, its solvate, or their inclusion compound, and (2) a saccharide, and optionally (3) a pH regulator Methyl 4-{[2-((1R, 2R, 3R) -3 in the lyophilized product], wherein the aqueous solution having a pH of 3 to 6.5 is subjected to lyophilization to obtain a lyophilized product. A method for stabilizing -hydroxy-2-{(1E, 3S) -3-hydroxy-4- [3- (methoxymethyl) phenyl] but-1-enyl} -5-oxocyclopentyl) ethyl] sulfanyl} butanoate;
[27] Prevention and treatment of an EP4-mediated disease in a mammal, comprising administering an effective amount of the composition described in [1], [2], [9] or [10] to the mammal And / or a method for inhibiting progression;
[28] Use of the composition according to the above [1], [2], [9] or [10] for producing a preventive, therapeutic and / or progression inhibitor for EP4-mediated diseases;
[29] Per unit form, (1) methyl 4-{[2-((1R, 2R, 3R) -3-hydroxy-2-{(1E, 3S) -3-hydroxy-4- [3- ( Methyl 4-{[2-((1R, 2R, 3R) -3-hydroxy-2-] containing 1 μg to 20 μg as methoxymethyl) phenyl] but-1-enyl} -5-oxocyclopentyl) ethyl] sulfanyl} butanoate An α-cyclodextrin inclusion compound of {(1E, 3S) -3-hydroxy-4- [3- (methoxymethyl) phenyl] but-1-enyl} -5-oxocyclopentyl) ethyl] sulfanyl} butanoate, and ( 2) Methyl 4-{[2-((1R, 2R, 3R) -3-hydroxy-2-{(1E, 3S) -3-hydroxy-4- [3- (methoxymethyl) fe [Lu] but-1-enyl} -5-oxocyclopentyl) ethyl] sulfanyl} butanoate is filled with a stable freeze-dried product having a water content of 1% or less and containing 1000 parts by weight to 10,000 parts by weight of lactose A vial of
[30] Per unit form, (1) methyl 4-{[2-((1R, 2R, 3R) -3-hydroxy-2-{(1E, 3S) -3-hydroxy-4- [3- ( Methoxy 4-{[2-((1R, 2R, 3R) -3-hydroxy-2-{((1R, 2R, 3R)]) containing 10 μg as methoxymethyl) phenyl] but-1-enyl} -5-oxocyclopentyl) ethyl] sulfanyl} butanoate 1E, 3S) -3-hydroxy-4- [3- (methoxymethyl) phenyl] but-1-enyl} -5-oxocyclopentyl) ethyl] sulfanyl} butanoate, an α-cyclodextrin inclusion compound, and (2) Methyl 4-{[2-((1R, 2R, 3R) -3-hydroxy-2-{(1E, 3S) -3-hydroxy-4- [3- (methoxymethyl) phenyl] buta −1-enyl} -5-oxocyclopentyl) ethyl] sulfanyl} butanoate A vial filled with a stable lyophilized product containing 5000 parts by weight of lactose and having a water content of 1% or less;
[31] (1) Methyl 4-{[2-((1R, 2R, 3R) -3-hydroxy-2-{(1E, 3S) -3-hydroxy-4- [3- (methoxymethyl) phenyl] Butane-1-enyl} -5-oxocyclopentyl) ethyl] sulfanyl} butanoate, its solvate, or their inclusion compounds, and (2) a saccharide, and optionally (3) a pH regulator An aqueous solution having a pH of 3 to 6.5; and [32] methyl 4-{[2-((1R, 2R, 3R) -3-hydroxy-2-{(1E, 3S) -3-hydroxy- The water according to [31] above, wherein the concentration of 4- [3- (methoxymethyl) phenyl] but-1-enyl} -5-oxocyclopentyl) ethyl] sulfanyl} butanoate is 3 μg / mL to 20 μg / mL. Liquid;
Etc.

  (1) Methyl 4-{[2-((1R, 2R, 3R) -3-hydroxy-2-{(1E, 3S) -3-hydroxy-4- [3- (methoxymethyl) phenyl] ] But-1-enyl} -5-oxocyclopentyl) ethyl] sulfanyl} butanoate, a solvate thereof, or an inclusion compound thereof, and (2) a saccharide, and optionally (3) a pH regulator. The stable pharmaceutical composition (hereinafter sometimes abbreviated as the composition of the present invention) containing methyl 4-{[2-((1R, 2R, 3R) -3-hydroxy, which is an active ingredient]. -2-{(1E, 3S) -3-hydroxy-4- [3- (methoxymethyl) phenyl] but-1-enyl} -5-oxocyclopentyl) ethyl] sulfanyl} butanoate (hereinafter 5-thia-PG Conversion And may be abbreviated as a compound)), a saccharide, and optionally a stable pharmaceutical composition containing a pH regulator, and other additives. It is not limited by the presence or absence of a solvent, the presence or absence of a solvent, the shape (solid, liquid) of a composition, etc.

  The composition of the present invention is stable. In particular, as described below using the terms “purity stability” and “morphological stability”, it is stable and excellent in both purity and form, so that it is used as a pharmaceutical preparation, particularly as an injectable preparation. It is preferable to use it. The form of the injectable preparation may be a liquid composition generally called a concentrated injection or concentrated injection, or a solid composition such as a lyophilized product. In the present specification, the preparation for injection means an injection precursor capable of preparing an injection that can be administered to a patient by using a solution and / or a diluted solution at the time of use.

  In the case where the composition of the present invention is solid, an injection can be prepared using a diluent after having once made a high-concentration liquid using a solution. Of course, a high-concentration liquid prepared using a solution is also a composition of the present invention. On the other hand, when the composition of the present invention is a liquid, an injection can be prepared by diluting it directly with a diluent. Furthermore, when the composition of the present invention is a liquid, it can be made into a solid composition (lyophilized product) of the present invention by subjecting the raw material to a treatment such as lyophilization. The composition of the present invention is preferably a solid composition, particularly a lyophilized product. In the present specification, the injection may be a liquid injection composition that can be administered parenterally directly to a patient. The injection prepared using the composition of the present invention may be in the form of, for example, an aqueous injection, a non-aqueous injection, a suspension injection, an emulsion injection, an infusion preparation and the like. The prepared injection can be applied to tissues such as intramuscular, intradermal, subcutaneous, intravenous, intraarterial, intraperitoneal, and spinal cord.

  In the present invention, methyl 4-{[2-((1R, 2R, 3R) -3-hydroxy-2-{(1E, 3S) -3-hydroxy-4- [3- (methoxymethyl) phenyl] buta- 1-enyl} -5-oxocyclopentyl) ethyl] sulfanyl} butanoate has the formula (I)

(Where

Represents the α-configuration,

Represents the β-configuration. )
It is a compound shown by these.

  Examples of the solvate of 5-thia-PG compound include solvates such as water and alcohol solvents (for example, ethanol). Solvates preferably have low toxicity and water solubility.

  The inclusion compound of 5-thia-PG compound or a solvate thereof means a compound in which 5-thia-PG compound or a solvate thereof becomes a guest compound and is included in the host compound. As the host compound, any compound that can include a 5-thia-PG compound or a solvate thereof can be used without particular limitation. For example, unimolecular host compounds (eg, cyclodextrins, crown compounds, cyclophanes, azacyclophanes, calixarenes, cyclotriperatrilenes, spherands, cavitands, cyclic oligopeptides, etc.), multimolecular host compounds (eg, Urea, thiourea, deoxycholic acid, perhydrotriphenylene, tri-o-thymotide, etc.), polymeric host compounds (eg, cellulose, starch, chitin, chitosan, polyvinyl alcohol, etc.), inorganic host compounds (eg, interlayers) Compound, zeolite, Hoffman complex, etc.). Of these host compounds, monomolecular host compounds, particularly hydrophilic monomolecular host compounds are preferred, and cyclodextrin is particularly preferred. Examples of cyclodextrins include α-cyclodextrin, β-cyclodextrin, and γ-cyclodextrin. In addition to these, some of these hydroxyl groups may be substituted with other functional groups such as alkyl groups, allyls. You may use the cyclodextrin derivative etc. which changed into group, an alkoxy group, an amide group, a sulfonic acid group, etc. As the host compound in the present invention, α-cyclodextrin is suitable.

  In the present invention, 5-thia-PG compounds, solvates thereof, or clathrate compounds thereof are not limited to those that are substantially pure and single substances, and are within the scope acceptable as an active pharmaceutical ingredient. If present, it may contain impurities (for example, a by-product derived from the production process, a solvent, a raw material, or a decomposition product). The content of impurities acceptable as an active pharmaceutical ingredient varies depending on the impurities contained therein. For example, in the α-cyclodextrin inclusion compound of 5-thia-PG compound, the amount of each related substance is about 1.0. %, The total amount of related substances is preferably about 4.0% or less, and the water content is preferably about 5.0% or less.

  The 5-thia-PG compound or a solvate thereof can be obtained by a method known per se, for example, the method described in WO 00/03980, Comprehensive Organic Transformations: A Guide to Functional Group Preparations, Second Edition (Richard C. Larock, John Wiley and Sons Inc., 1999) [Comprehensive Organic Transforms: A Guide to Fundamental Production & Lr. Preparations, 2nd Edition, 2nd Ed. Inc., 1999)] alone or in combination. It can be produced by there. In addition, the produced 5-thia-PG compound or a solvate thereof is obtained by a conventional purification means, for example, distillation under normal pressure or reduced pressure, high performance liquid chromatography using silica gel or magnesium silicate, thin layer chromatography. Alternatively, it can be purified by column chromatography or a method such as washing and recrystallization.

  An inclusion compound of a 5-thia-PG compound or a solvate thereof and a host compound is obtained by subjecting a 5-thia-PG compound or a solvate thereof to a host compound by a method known per se, for example, publication of a British patent application. It can be produced by subjecting it to the methods described in the specification of No. 1351238 and British Patent Application No. 1419221.

  In the present invention, the saccharide can be used without particular limitation as long as it is a saccharide generally used in the production of injectable preparations. For example, glucose, galactose, mannose, lactose, maltose, palatinose, trehalose, raffinose, erulose, melecitose and the like are preferable, for example, lactose, maltose and the like are more preferable, for example, maltose and the like are particularly preferable. These saccharides may be crystalline or non-crystalline, and may be hydrated or anhydrous. In particular, if these sugars listed in the Japanese Pharmacopoeia, for example, lactose, lactose monohydrate (4-O-β-Galactopyranosyl-α-D-glucopyranose monohydrate) is maltose, Maltose monohydrate (4-O-α-D-Glucopyranosyl-β-D-glucopyranose monohydrate) is preferred. In addition, maltose monohydrate may be referred to as crystalline maltose. Maltose having a pH of 4.5 to 6.5 means maltose having a pH of 4.5 to 6.5 in a solution obtained by dissolving 1.0 g of maltose in 10 mL of water. For example, maltose (maltose monohydrate) listed in the Japanese Pharmacopoeia is included in maltose having a pH of 4.5 to 6.5.

  The amount of saccharide added in the composition of the present invention is not particularly limited. For example, the amount is preferably about 100 parts by weight to about 100,000 parts by weight with respect to 1 part by weight of 5-thia-PG compound. More preferred is 500 parts by weight to about 50000 parts by weight, and particularly preferred is about 1000 parts by weight to about 10,000 parts by weight. In particular, it is preferable to add about 5000 parts by weight of saccharide to 1 part by weight of 5-thia-PG compound. In addition, these saccharides may be used alone or in combination of two or more.

  In the present invention, the pH adjuster can be used without particular limitation as long as it is a pH adjuster generally used in the production of injectable preparations. For example, organic acids such as citric acid, tartaric acid, oxalic acid, acetic acid, lactic acid (especially divalent or trivalent organic acids (for example, citric acid, tartaric acid, oxalic acid, etc.)), for example, hydrochloric acid, phosphoric acid, etc. Inorganic acids such as sodium hydroxide, potassium hydroxide, lithium hydroxide and other metal hydroxides (especially alkali metal or alkaline earth metal hydroxides) such as sodium citrate, potassium tartrate, acetic acid Organic acid metal salts such as sodium (especially alkali metal or alkaline earth metal salts of divalent or trivalent organic acids) and the like are preferable. These pH regulators may be used alone or in combination of two or more. In the present invention, for example, organic acid metal salts, organic acids, metal hydroxides, or combinations thereof are more preferable. For example, organic acid metal salts (for example, sodium citrate) and organic acids (for example, A combination of a metal hydroxide (for example, sodium hydroxide) and an organic acid (for example, citric acid) is particularly preferable. In particular, a combination of a metal hydroxide (for example, sodium hydroxide) and an organic acid (for example, citric acid) is preferable.

  The addition amount of the pH regulator in the composition of the present invention is not particularly limited, but in order to make the composition of the present invention a stable composition, for example, when the composition of the present invention is a liquid, the pH is about 3 It is preferable to add so that it is within the range of about 6.5 to about 6.5, preferably about 3.5 to about 5, particularly preferably about 4. Also, for example, when the composition of the present invention is a solid, the concentration of 5-thia-PG compound is about 1 μg / mL to about 50 μg / mL, more preferably about 3 μg / mL to about 20 μg / mL, particularly preferably When dissolved in water for injection to a pH of about 16 μg / mL to about 17 μg / mL (eg, 10 μg / 0.6 mL), the pH is about 3 to about 6.5, preferably about 3.5 to about It is preferred to add such an amount that is in the range of 5, particularly preferably about 4.

  In addition to the saccharides and pH adjusters described above, other additives generally used in the manufacture of injectable preparations may be appropriately added to the composition of the present invention. Examples of such additives include 70 v / v% N-hydroxyethyl lactamide aqueous solution, D-sorbitol, D-mannitol, DL-methionine, L-aspartic acid, L-alanine, L-arginine, L-glutamic acid. , L-lysine, potassium L-glutamate, sodium L-glutamate, L-cystine, L-cysteine, L-histidine, L-methionine, N, N-dimethylacetamide, ascorbic acid, acetyltryptophan sodium, aminoethylsulfone Acid, aminoacetic acid, gum arabic, gum arabic powder, alpha thioglycerin, albumin, inositol, ethanol, ethyl urea, ethylenediamine, calcium sodium edetate, sodium edetate, oleic acid, sodium caprylate, carmellosna Lithium, xylitol, disodium citrate, glycerin, calcium gluconate, sodium gluconate, magnesium gluconate, creatinine, chlorobutanol, gentisic acid ethanolamide, succinic acid, sesame oil, sodium chondroitin sulfate, sodium salicylate, diethanolamine, diethylenetriaminepentaacetic acid Sorbitan sesquioleate, gelatin, gelatin hydrolyzate, sorbitan fatty acid ester, soybean oil, thioglycolic acid, sodium thioglycolate, potassium thiocyanate, sodium thiomalate, sodium thiosulfate, camellia oil, dextran 40, dextran 70, Sodium desoxycholate, triethanolamine, sodium formaldehyde, sulfoxylate, nicotinic acid Mido, ethyl paraoxybenzoate, butyl paraoxybenzoate, propyl paraoxybenzoate, methyl paraoxybenzoate, hydroxypropylcellulose, castor oil, potassium pyrosulfite, sodium pyrosulfite, phenol, butylhydroxyanisole, propylene glycol, sodium heparin, benzyl Alcohol, polyoxyethylene (160) polyoxypropylene (30) glycol, polyoxyethylene castor oil, polyoxyethylene hydrogenated castor oil, polyoxyethylene hydrogenated castor oil 50, polyoxyethylene hydrogenated castor oil 60, polysorbate 80, macrogol 400, macrogol 4000, meglumine, sodium methanesulfobenzoate, monoethanolamine, aluminum monostearate, monolaurin Acid polyoxyethylene sorbitan (20E.O.), peanut oil, sodium sulfite, sodium hydrogen sulfite, benzoic acid, sodium benzoate, benzyl benzoate, aluminum chloride, sodium chloride, benzalkonium chloride, benzethonium chloride, magnesium chloride, Zinc chloride, zinc chloride solution, stannous chloride, ferric chloride, arginine hydrochloride, cysteine hydrochloride, lysine hydrochloride, fructose, dried aluminum gel, dried sodium sulfite, dilute hydrochloric acid, highly purified egg yolk lecithin, calcium oxide, zinc oxide, odor Calcium chloride, sodium bromide, ammonium acetate, sodium acetate, zinc acetate, aluminum hydroxide, purified gelatin, purified soybean lecithin, purified soybean oil, purified sucrose, purified egg yolk lecithin, water for injection, calcium carbonate, lactic acid, ethyl lactate, Lactated sodium Liquid, urea, concentrated glycerin, sodium absolute ethanol, pyrophosphate anhydride, maleic anhydride, stannous chloride, anhydrous sodium acetate, sulfate, aluminum potassium sulfate, potassium sulfate, magnesium sulfate and the like. These additives are generally blended in a proportion usually used for injectable preparations. Moreover, these additives may be used independently and may be used in combination of 2 or more types. It is easy for those skilled in the art, and these additives may be used according to the purpose of use, as described in “Pharmaceutical Additives Dictionary” (edited by Japan Pharmaceutical Additives Association) published in 2000. E.g. stabilizers, surfactants, buffers, solubilizers, antioxidants, antifoaming agents, isotonic agents, emulsifiers, suspending agents, preservatives, soothing agents, solubilizers, solubilizers It can be properly used as an auxiliary agent.

  The addition amount of other additives in the composition of the present invention is not particularly limited, but in order to give preferable stability to the composition of the present invention, the addition amount is determined using the same index as the above-mentioned pH regulator. Just do it. Specifically, for example, when the composition of the present invention is a liquid, the pH is in the range of about 3 to about 6.5, preferably about 3.5 to about 5, particularly preferably about 4. It is preferable to add. Also, for example, when the composition of the present invention is a solid, the concentration of 5-thia-PG compound is about 1 μg / mL to about 50 μg / mL, more preferably about 3 μg / mL to about 20 μg / mL, particularly preferably When dissolved in water for injection to a pH of about 16 μg / mL to about 17 μg / mL (eg, 10 μg / 0.6 mL), the pH is about 3 to about 6.5, preferably about 3.5 to about It is preferred to add such an amount that is in the range of 5, particularly preferably about 4.

  The pH adjusting agent and other additives are optionally added to the composition of the present invention. If the 5-thia-PG compound is stable only by adding a saccharide, the pH adjusting agent and other additives are added. Addition of the agent is not essential. Whether or not a pH adjusting agent or other additives are added can be determined as follows. For example, when the composition of the present invention comprising a 5-thia-PG compound and a saccharide is a liquid, the pH is from about 3 to about 6.5, preferably from about 3.5 to about 5, particularly preferably about 4. If it is within the range, addition of a pH adjusting agent and other additives is not essential. For example, when the composition of the present invention containing a 5-thia-PG compound and a saccharide is a solid, the concentration of the 5-thia-PG compound is about 1 μg / mL to about 50 μg / mL, more preferably about 3 μg. PH is about 3 to about 6 when dissolved with water for injection so as to be about 16 μg / mL to about 17 μg / mL (for example, 10 μg / 0.6 mL). 0.5, preferably from about 3.5 to about 5, particularly preferably in the range of about 4, the addition of pH adjusters and other additives is not essential. Specifically, as described in the Examples below, when the composition of the present invention which is an aqueous solution is prepared using lactose as the saccharide, the pH is in the range of about 4 to about 5, so that the pH adjuster The addition of other additives is not essential.

  As described above, the composition of the present invention includes a solid (for example, freeze-dried product) and a liquid (for example, an aqueous solution for producing a freeze-dried product or an aqueous solution in which a freeze-dried product is redissolved). However, the degree of stability differs depending on the two, and the production method and preferred use form thereof are different. Hereinafter, the case of an aqueous solution (particularly an aqueous solution for producing a lyophilized product) and the case of a lyophilized product will be specifically described as examples. It may be abbreviated as “dried product”.

  The aqueous solution of the present invention is prepared by adding (1) a 5-thia-PG compound, a solvate thereof, or an inclusion compound thereof, and (2) a saccharide to water, which is a solvent, and (3) a pH if desired. It is prepared by adding a regulator and (4) other additives. In the preparation of the aqueous solution of the present invention, addition and mixing operations are carried out according to ordinary pharmaceutical methods. For example, an aqueous solution containing (1) a 5-thia-PG compound, a solvate thereof, or an inclusion compound thereof, (2) a saccharide, (3) a pH regulator, and (4) other additives is prepared. When weighing, each may be weighed and mixed and then dissolved in water, or after weighing and dissolved in water without mixing. The order of addition is not particularly limited. It is also possible to prepare an aqueous solution of the present invention by preparing an aqueous solution in which each component is dissolved separately and mixing the aqueous solutions. In addition, when using an inclusion compound of 5-thia-PG compound or a solvate thereof as a solute, it is not necessary to use a compound prepared beforehand as an inclusion compound of 5-thia-PG compound or a solvate thereof. A 5-thia-PG compound or a solvate thereof and a host compound can be added separately to form an inclusion compound in an aqueous solution. That is, for example, even a 5-thia-PG compound, a host compound, a saccharide, and optionally a pH adjuster separately added to water as a solvent are included in the aqueous solution of the present invention. Needless to say, the freeze-dried product prepared as described below using the aqueous solution of the present invention is included in the freeze-dried product of the present invention.

  The concentration of the solute in the aqueous solution of the present invention is not particularly limited, but the concentration of the 5-thia-PG compound in the aqueous solution is preferably, for example, about 1 μg / mL to about 50 μg / mL, and about 3 μg / mL to about 20 μg / mL. mL is more preferred. In particular, about 16 μg / mL to about 17 μg / mL (for example, 10 μg / 0.6 mL) is preferable. Preferable amounts of addition of sugars, pH regulators and other additives are as described above.

  The aqueous solution of the present invention is stable and stable not only in its form but also in terms of purity. In other words, it is excellent not only in form stability that there is no change in appearance over time but also in purity stability. Purity stability can be shown, for example, using an index called a residual rate. The residual ratio refers to the ratio of the amount of 5-thia-PG compound after storage to the amount of 5-thia-PG compound before storage. The aqueous solution of the present invention has, for example, a residual ratio of 5-thia-PG compound of about 80% or more even after storage for 24 hours under conditions of 25 ° C. and 60% relative humidity (RH). 80% to about 100%), preferably about 85% or more (about 85% to about 100%), more preferably about 90% or more (about 90% to about 100%), particularly preferably about 93% or more (about 93% or more to about 100%) (purity stability). An aqueous solution having a particularly high residual ratio, for example, a residual ratio of 5-thia-PG compound of about 99% or more (about 99% to about 100% after storage for 24 hours under conditions of 25 ° C. and 60% relative humidity (RH)) ) To adjust the pH to about 4 to about 5.5 using crystalline maltose, citric acid and sodium citrate as additives, or (2) crystalline maltose and citric acid Is used as an additive to adjust the pH to about 3.5 to about 4.5, or (3) the pH is about 4 to about 5 using crystalline maltose, citric acid and sodium hydroxide as additives. Adjust it. Further, when lactose is used as an additive, the pH is in the range of about 4 to about 5, so that the same effect can be obtained without adding a pH adjuster.

  Since the aqueous solution of the present invention is stable, it is sterilized as it is, filled into an appropriate injection container, and if desired, applied to a light-shielding package (for example, an injection container containing the freeze-dried product of the present invention). It can be used as a concentrated injection by applying light-shielding packaging, etc., which will be described later as packaging, but it should be freeze-dried to obtain a freeze-dried product in order to obtain a stable injectable preparation for a long period of time. Is preferred. In particular, the aqueous solution of the present invention does not rapidly decompose the active ingredient 5-thia-PG compound even if it remains in the aqueous solution until it is freeze-dried after preparation. High aqueous solution can be provided.

  The freeze-dried product of the present invention can be produced by filling the aqueous solution of the present invention into an injection container and subjecting it to freeze-drying. The filling amount of the aqueous solution of the present invention into the injection container is not particularly limited, but for example, about 0.5 mL to about 1.5 mL, more preferably about 0.5 mL to about 1.0 mL, particularly preferably about 0.6 mL. Or it is preferable to fill about 1.0 mL, especially about 0.6 mL of aqueous solution. The content of 5-thia-PG compound per unit for injection (for example, 1 vial, 1 ampoule, etc.) is not particularly limited, but for example, about 0.1 μg to about 20 μg, preferably about 1 μg to about 20 μg, More preferred is a content of about 1 μg to about 12 μg, and particularly preferred is a content of about 2 μg, about 5 μg, about 6 μg, or about 10 μg. In addition, it can be set as the injection container which hold | maintained sterility by giving to the sterilization operation similar to a general injection ampoule, a vial, a syringe, etc. in the arbitrary processes in these processes. Further, if desired, before filling into these containers, an operation such as filtration with a dustproof filter (for example, 0.45 μm methylcellulose membrane, 0.45 μm nylon 66 membrane, 0.45 μm polyvinylidene fluoride membrane, etc.) may be performed. Good. Specific sterilization methods used for the sterilization operation include, for example, gas sterilization method, hot water immersion sterilization method, hot water shower sterilization method, filtration sterilization method, irradiation sterilization method (for example, electron beam sterilization method, ultraviolet sterilization method, γ-ray sterilization method), high-pressure steam sterilization (autoclave) method and the like. The filtration sterilization method is, for example, after preparing the aqueous solution of the present invention by the above-described method and before filling it into an appropriate container such as an ampoule, vial, syringe, etc. Membrane, 0.22 μm nylon 66 membrane, 0.22 μm polyvinylidene fluoride membrane, etc.). The gas sterilization method is performed on injection containers such as ampoules, vials, and syringes before filling with the aqueous solution of the present invention. Hot water immersion sterilization method, hot water shower sterilization method, irradiation sterilization method and high-pressure steam sterilization method, for example, prepare the aqueous solution of the present invention by the above-mentioned method, and fill it into appropriate containers such as ampoules, vials, syringes, etc. Done after. The high-pressure steam sterilization is preferably performed, for example, at 100 ° C. to 125 ° C. for 5 minutes to 30 minutes. As the sterilization method in the present invention, for example, a filtration sterilization method, a high-pressure steam sterilization method, and the like are preferable, and a filtration sterilization method (a filtration sterilization method using a filter having a pore diameter of 0.22 μm) is particularly preferable.

  In the present invention, the injection container may be in any form as long as it is sealable and can maintain the sterility of the contents, but generally it is filled with an injectable preparation or injection. Vials and ampoules used in the above are preferred. It is also possible to fill a syringe with a lyophilized solution (for example, distilled water for injection or physiological saline) to obtain a prefilled syringe (double chamber type syringe). These injection containers may be made of any material. For example, glass and plastic materials are preferable for vials and ampoules, and plastic materials are preferable for double-chamber syringes. . Further, by combining these materials, for example, a glass container whose inner surface is coated with a plastic material or a plastic container whose inner surface is coated with a glass material can be used.

  The glass material should just be a glass material which can be used for the container for a pharmaceutical filling. When a glass container is used, the inner surface of the container may be coated with silicon or treated with silicon dioxide. Silicon coating is performed using a silicon-based coating agent (for example, dimethyl silicone oil, methyl phenyl silicone oil, methyl hydrogen silicone oil, etc.) and the like, and the thickness of the coating is about 100 μm or less, preferably It is performed by subjecting to a known method, for example, a heat deposition method, a plasma enhanced chemical vapor deposition method, a plasma pulse chemical vapor deposition method, or the like so as to be about 15 μm to about 50 μm or less. The treatment of silicon dioxide is carried out by subjecting it to a known method such as a silicote treatment, a wave plasma chemical vapor phase treatment or the like. In addition, glass containers whose inner surfaces have been treated with silicon dioxide in advance (for example, silicote ampoules, silicote vials (made by Shioya Glass, Fuji glass), type I plus ampules, type I plus vials (made by SCHOTT), etc.) It may be used.

  The plastic material may be any plastic material that can be used for a container for filling pharmaceutical products, for example, polyethylene, polypropylene, polystyrene, polymethylpentene, polyester, polyamide, polycarbonate, or a cyclic olefin compound or a crosslinked polycyclic hydrocarbon. A polymer resin of a compound can be used without particular limitation.

  When an ampule is used as an injection container, the ampule is filled with the aqueous solution of the present invention, freeze-dried, and then sealed by sealing the opening.

  When a vial is used as an injection container, such a vial is filled with the aqueous solution of the present invention, subjected to freeze-drying, and then sealed by combining a rubber stopper with an opening and, if desired, an aluminum cap.

  When a syringe (double-chamber type syringe) is used as an injection container, the syringe is filled with the aqueous solution of the present invention in one chamber (for example, the tip side of the syringe) and subjected to lyophilization, and then the remaining chamber. One chamber (for example, the plunger rod side) is filled with a lyophilized solution (for example, distilled water for injection or physiological saline), and the tip for attaching the injection needle is made of rubber or Sealing is performed by sealing with a plastic part and sealing the plunger rod with a rubber or plastic gasket or plunger rod.

  A preferable injection container used for filling the aqueous solution of the present invention includes a vial. In particular, a glass vial is preferable, and a glass vial whose inner surface is coated with silicon is particularly preferable.

  Freeze-drying is performed by the steps of preliminary freeze-drying, vacuum primary drying and secondary drying. The pre-lyophilization is performed by freezing at about −50 ° C. to about −40 ° C. for about 2 hours to about 5 hours, preferably about −50 ° C. to about −45 ° C. for about 2.5 hours to about 4.5 hours. Done. Vacuum primary drying is a process performed subsequent to preliminary freeze-drying. In the primary vacuum drying, the shelf temperature is raised to about 0 ° C. to about 20 ° C. under the condition of a vacuum degree of about 5 Pascals to about 20 Pascals, and the temperature is about 5 hours to about 10 hours, preferably about 0 ° C. The temperature is raised to about 15 ° C., and vacuum drying is performed at that temperature for about 6 hours to about 9 hours. Secondary drying is a process performed subsequent to vacuum primary drying. Secondary drying is performed by raising the temperature from about 0 ° C. to about 20 ° C. to about 25 ° C. to about 55 ° C., preferably from about 35 ° C. to about 50 ° C., optionally from about 5 ° C. to about 15 ° C. The drying step is preferably performed at a temperature of about 10 ° C. to about 15 ° C. and is performed for about 7 hours to about 10 hours, preferably about 7.5 hours to about 9.5 hours.

  The injection container containing the freeze-dried product of the present invention thus produced is provided with a light-shielding package as desired. Such a packaging can be used without particular limitation as long as it is a light-shielding packaging that is generally used. Specifically, a bag made of a material that suppresses the transmission of light of a specific wavelength, a bag made of a light shielding material such as plastic or aluminum, or a shrink package (for example, a shrink label) or a blister package using a light shielding plastic. Etc. can be used. These light-shielding packagings can be further improved in light-shielding properties by being used in combination.

  Since the 5-thia-PG compound used in the present invention is a substance unstable to light, it is preferable to provide a light-shielding package for the injection container containing the lyophilized product of the present invention. . For example, it can be shielded from light by putting it in a box or the like, but it is particularly preferable to apply shrink packaging (for example, a shrink label) using light-shielding plastic.

  The lyophilized product of the present invention preferably has a low water content, for example, about 2% or less, preferably about 1% or less, more preferably about 0.8% or less, particularly preferably about 0.5% or less. Those having a water content are preferred. Of course, in the freeze-dried product of the present invention, it is preferable that the water content is low, and it is particularly preferable that the water content is substantially free, ie, below the detection limit. In the lyophilized product of the present invention, the residual ratio of the 5-thia-PG compound can be increased by lowering the water content.

  The freeze-dried product of the present invention is stable and excellent in purity stability. The lyophilized product of the present invention has a residual ratio of 5-thia-PG compound of about 70% or more (about 70% to about 70% to about 70% or more) even when the lyophilized product is produced and stored at 60 ° C. for 2 weeks. 100%), preferably about 75% or more (about 75% to about 100%), more preferably about 80% or more (about 80% to about 100%), particularly preferably about 85% or more (about 85% to about 100%). 100%). That is, since the 5-thia-PG compound which is an active ingredient is not rapidly decomposed after the lyophilized product of the present invention is manufactured, a lyophilized product having a high residual rate can be provided as an injectable preparation for clinical use. Is possible. A freeze-dried product having a particularly high residual rate, for example, a freeze-dried product having a residual rate of 5-thia-PG compound of about 95% or more (about 95% to about 100%) after storage for 2 weeks at 60 ° C. (1) Crystalline maltose, citric acid and sodium citrate are added in combination as additives, and the pH is about 4 to about 4.5, preferably about 4 to about 4.3, particularly preferably about 4 The lyophilized product is prepared using the aqueous solution of the present invention adjusted to about 4.1, or (2) crystalline maltose, citric acid and sodium hydroxide are added in combination as an additive, and the pH is about 4 to about A freeze-dried product may be produced using the aqueous solution of the present invention adjusted to 5, preferably about 4 to about 4.6.

  The freeze-dried product of the present invention is also excellent in form stability. The freeze-dried product of the present invention is substantially the same as before storage (initial stage), for example, after the freeze-dried product is manufactured, and does not substantially change in shape even when stored at about 60 ° C. for about 2 weeks, preferably about 4 weeks. The same (morphological stability). Shape change refers to appearance changes such as shrinkage, melting, deliquescence and / or dissolution of a freeze-dried product over time, for example, initially spongy, cottony and / or spongey, white or extremely The lyophilized cake showing a light pale yellow color tone may be crushed and changed into a yellow color tone that is crushed and / or semi-dissolved over time.

  Hereinafter, a more specific method for producing the freeze-dried product of the present invention will be described.

  The lyophilized product of the present invention is prepared by, for example, adding an α-cyclodextrin inclusion compound of 5-thia-PG compound and a saccharide (for example, crystalline maltose, etc.) in an appropriate amount of distilled water for injection according to a usual pharmaceutical method. Dissolving, adding a pH adjusting agent (eg, citric acid and sodium hydroxide) to adjust the pH to about 3 to about 6.5, preferably about 3.5 to about 5, particularly preferably about 4, and After adding an appropriate amount of distilled water for injection to adjust the liquid volume, the solution is sterilized by filtration using a sterilization filter or the like, and the above-mentioned injection container (preferably, a glass vial or the like whose inner surface is coated with silicon) is 1 The 5-thia-PG compound per unit form is, for example, about 0.1 μg to about 20 μg, preferably about 1 μg to about 20 μg, more preferably about 1 μg to about 12 μg, particularly preferably about 2 μg, about 5 μg, about [mu] g, was filled to contain about 10 [mu] g, then subjected to freeze-drying of the can be produced by sealing. In addition, the unit form here means the form used as the unit at the time of dispensing, such as 1 vial, 1 ampule, etc., for example.

  In the present invention, the method for stabilizing 5-thia-PG compound is a method for suppressing the decomposition of 5-thia-PG compound in the freeze-dried product of the present invention, and derived from 5-thia-PG compound. This is a method for suppressing the generation of decomposition products. Specifically, (1) adding 5-thia-PG compound used for the production of the lyophilized product as an inclusion compound with α-cyclodextrin, (2) pH of the aqueous solution used for the production of the lyophilized product Saccharide and optionally pH adjusting agent to adjust to about 3 to about 6.5, preferably about 3.5 to about 5, particularly preferably about 4, and / or (3) lyophilization This is done by lowering the moisture content of the product. By satisfying any of these, preferably all, it is possible to more effectively suppress the decomposition of the 5-thia-PG compound in the lyophilized product.

  The composition of the present invention provides methyl 4-{[2-((1R, 2R, 3R) -3-hydroxy-2-{(1E, 3S) -3-hydroxy-] associated with storage of 5-thia-PG compounds. Suppressed production of 4- [3- (methoxymethyl) phenyl] but-1-enyl} -5-oxocyclopentyl) ethyl] sulfanyl} butanoate degradation product (hereinafter sometimes simply referred to as degradation product) It is. Here, as the decomposition product, for example, the formula (IA)

(Wherein all symbols have the same meaning as described above), formula (IB)

(Wherein all symbols have the same meaning as described above), formula (IC)

(Wherein all symbols have the same meaning as described above), formula (ID)

(Wherein all symbols have the same meaning as described above), formula (IE)

(Wherein all symbols have the same meaning as described above), and the like. In particular, the compounds represented by formula (IA) and formula (IB) are the main decomposition products. The compounds represented by the formulas (IA), (IB), (IC), (ID), and (IE) are methyl 4-{[2-(( 1R, 2S) -2-{(1E, 3S) -3-hydroxy-4- [3- (methoxymethyl) phenyl] but-1-enyl} -5-oxocyclopent-3-en-1-yl) Ethyl] sulfanyl} butanoate (IA), methyl 4-{[2-((1R, 2R, 3R) -3-hydroxy-2-{(1E, 3S) -3-hydroxy-4- [3- ( Methoxymethyl) phenyl] but-1-enyl} -5-oxocyclopentyl) ethyl] sulfinyl} butanoate (IB), methyl 4-{[2-((1R, 2R, 3R) -3-hydroxy-2- {(1E, 3S) -3-hydroxy-4- [3- (methoxy Methyl) phenyl] but-1-enyl} -5-oxocyclopentyl) ethyl] sulfonyl} butanoate (IC), methyl 4-{[2-((1R, 2S) -2-{(1E, 3S)- 3-hydroxy-4- [3- (methoxymethyl) phenyl] but-1-enyl} -5-oxocyclopent-3-en-1-yl) ethyl] sulfinyl} butanoate (ID), methyl 4- {[2-((1R, 2S) -2-{(1E, 3S) -3-hydroxy-4- [3- (methoxymethyl) phenyl] but-1-enyl} -5-oxocyclopent-3- En-1-yl) ethyl] sulfonyl} butanoate (IE). These names are obtained by using a computer program that mechanically generates an IUPAC name, ACD / NAME ^™ (Advanced Chemistry Development).

  Since the composition of the present invention, particularly the lyophilized product, can reduce the production of the degradation product as much as possible, it is possible to provide an injectable preparation containing a 5-thia-PG compound having a high residual rate in the clinical field.

  For the residual ratio of the 5-thia-PG compound in the aqueous solution of the present invention and the freeze-dried product of the present invention, a known analysis method (for example, high performance liquid chromatography, gas chromatography, thin layer chromatography, etc.) was used. Although it can be calculated based on the measurement result, it is particularly preferable to perform measurement using a high performance liquid chromatographic method. The high performance liquid chromatographic method is performed by a known method. Specifically, it can be carried out by the method described in Examples described later. By using this method, the residual ratio of 5-thia-PG compound can be calculated by measuring the peak area of 5-thia-PG compound before and after storage.

  As described above, in the freeze-dried product of the present invention, the generation of decomposition products can be suppressed by reducing the water content. The water content in the lyophilized product can be measured using a known analysis method (for example, Karl Fischer method). Specifically, it can be carried out by the method described in Examples described later.

[Application to pharmaceutical products]
Since the composition of the present invention, particularly the lyophilized product contains a 5-thia-PG compound having EP4 activity, mammals (eg, humans, non-human animals such as monkeys, sheep, cows, It can be used as an injection for the purpose of preventing, treating and / or suppressing progression of EP4-mediated diseases in horses, dogs, cats, rabbits, rats, mice and the like. Here, the EP4-mediated disease may be any disease in which EP4 is involved in the process of formation, establishment, exacerbation, etc. of the disease or in the healing process. For example, an immune disease (for example, an autoimmune disease) (Examples: amyotrophic lateral sclerosis (ALS), multiple sclerosis, Sjogren's syndrome, rheumatoid arthritis, systemic lupus erythematosus, etc.), rejection after organ transplantation), asthma, neuronal cell death, lung injury, Liver disorder, acute hepatitis, nephritis, renal failure, hypertension, myocardial ischemia, systemic inflammatory response syndrome, burn pain, sepsis, hemophagocytic syndrome, macrophage activation syndrome, Still's disease, Kawasaki disease, burn, whole body Granuloma, ulcerative colitis, Crohn's disease, hypercytokinemia during dialysis, multiple organ failure, shock, sleep abnormalities, platelet aggregation, bone disease (eg, bone dysplasia, bone loss disease) ), And the like. Of these, ulcerative colitis and bone diseases, particularly bone loss diseases are suitable for the application of the composition of the present invention, particularly freeze-dried products. Incidentally, the bone mass reduction disease refers to a disease in which a bone mass reduction accompanied by symptoms such as bone density reduction and bone tissue degradation occurs. For example, (1) primary osteoporosis (for example, primary osteoporosis associated with aging) Osteoporosis, primary osteoporosis associated with menopause, primary osteoporosis associated with ovariectomy, vertebral fracture, etc., (2) secondary osteoporosis (eg, glucocorticoid-induced osteoporosis, hyperthyroid osteoporosis, fixation-induced osteoporosis) , Heparin-induced osteoporosis, immunosuppression-induced osteoporosis, osteoporosis due to renal failure, inflammatory osteoporosis, osteoporosis associated with Cushing syndrome, rheumatic osteoporosis, etc.), (3) cancer bone metastasis, hypercalcemia, Paget's disease, bone Bone diseases such as defects (alveolar bone defects, mandibular bone defects, sudden childhood bone defects, etc.), osteonecrosis, etc. can be mentioned. Furthermore, the composition of the present invention, particularly the lyophilized product, can be used for bone formation after bone surgery (eg, bone formation after fracture, bone formation after bone transplantation, bone formation after artificial joint surgery, post spinal fusion) Can be used as an injection for the purpose of accelerating / promoting healing of bone formation and other bone formation after bone reconstruction. It can also be used for bone graft replacement therapy.

  Among the above-mentioned bone loss diseases, vertebral fractures are mentioned as the most preferable diseases for the application of the composition of the present invention, particularly the freeze-dried product.

  The composition of the present invention, particularly a lyophilized product, is in a form that can be administered to a patient using a solution and / or diluent for the purpose of preventing, treating, and / or suppressing the progression of the above diseases, ie, an injection and And then administered in vivo. Here, the solution and / or diluent may be any aqueous solution that can be injected into a living body, preferably intravenously, such as distilled water for injection, physiological saline, sugar solution, and other general infusion solutions.

  The daily dose of the injection prepared using the composition of the present invention, particularly a freeze-dried product, varies depending on the degree of symptoms; age, sex, body weight of administration subject; timing of administration, interval, etc., and is not particularly limited. For example, when used as an injection for the purpose of prophylaxis, treatment and / or inhibition of progression of EP4-mediated diseases, the daily dose is about 4 ng to about 0.36 μg of 5-thia-PG per kg of patient body weight. It is preferred to administer the compound. In particular, when administered intravenously as a therapeutic agent for vertebral fractures, it is preferable to administer about 0.12 μg of 5-thia-PG compound per kg of the patient's body weight as a daily dose. As a more specific administration method in the case of intravenous infusion as a therapeutic agent for vertebral fractures, for example, about 0.12 μg of 5-thia-PG compound per 1 kg of a patient's body weight is divided twice a day. And a method of instilling over two hours each in the morning and afternoon.

  Injectables prepared using the composition of the present invention, particularly lyophilized products, include other drugs such as phosphodiesterase 4 inhibitor, bisphosphonate preparation, vitamin D preparation, calcium preparation, estrogen preparation, calcitonin preparation, isoflavone preparation, protein Anabolic steroid, vitamin K preparation, cathepsin K inhibitor, HMG-CoA reductase inhibitor, parathyroid hormone, growth factor, caspase-1 inhibitor, PTHrP derivative, metalloproteinase inhibitor, farnesoid X receptor agonist, It may be used in combination with estrogen agonist, progesterone agonist and the like. The administration method of these drugs used in combination is not particularly limited, and may be oral administration or parenteral administration.

  Examples of the phosphodiesterase inhibitor (PDE4) include cilomilast, roflumilast, allofylline, OPC-6535, ONO-6126, IC-485, AWD-12-281, CC-10004, CC- 1088, KW-4490, Iirimilast, ZK-117137, YM-976, BY-61-9987, CC-7085, CDC-998, MEM-1414, ND-1251, Bay19-8004, D-4396, PD-168787, Examples include Atizolam, Cipamfylline, Rolipram, NIK-616, SCH-351591, and V-11294A.

  Bisphosphonate preparations include, for example, alendronate sodium hydrate, ibandronic acid, incadronate disodium, etidronate disodium, olpadronate, Clodronate sodium hydrate, zoledronic acid, tiludronate disodium, neridronate, pamidronate disodium, pyridronate, minodronic acid water Examples include Japanese (Minodronic acid hydrate), risedronate sodium hydrate, YM175, and the like.

  Examples of vitamin D preparations include alphacalcidol, falecalcitriol, calcitriol, 1α, 25-dihydroxycholecalciferol, dihydrotaxosterol, ST-630, KDR, ST-630, ED-71, locartol (Ro44 -7190), tacalcitol, maxacalcitol and the like.

  Examples of calcium preparations include calcium chloride, calcium gluconate, calcium glycerophosphate, calcium lactate, calcium L-aspartate, and phosphoric acid. Examples include dibasic calcium phosphate.

  Estrogen preparations include, for example, Estradiol, Estradiol benzoate, Estradiol cypionate, Estradiol dipropionate, Estradiol enanthate, Estradiol hexahydrobenzoate. Art (Estradiol hexahydrobenzoate), Estradiol phenylpropionate, Estradiol undecanoate, Estradiol valerate, Estrone, Ethynyl estradiol, Mestranol, etc. Can be mentioned.

  Examples of the calcitonin preparation include, for example, calcitonin, salmon calcitonin (STH-32, SMC20-51), chicken calcitonin (MCI-536), cecalciferol, elcatonin, Examples include TJN-135.

  Examples of the isoflavone preparation include ipriflavone and the like.

  Anabolic steroids include, for example, oxymetholone (Oxymetholone), stanozolol (Stanozolol), nandrolone decanoate (Nandrolone phenylpropionate), nandrolone cyclohexylpropionate (Nandrolone cyclohexylpropionate), metenolone acetate (Metenolone acetate) acetate), mestanolone (Mestanolone), ethylestrenol (Ethylestrenol), calsterone (Calusterone) and the like.

  Examples of vitamin K preparations include menatetrenone and phytonadione.

  Examples of cathepsin K inhibitors include ONO-5334.

  Examples of HMG-CoA reductase inhibitors include pravastatin (sodium) (pravastatin (sodium)), simvastatin (simvastatin), fluvastatin (sodium) (fluvastatin (sodium)), cerivastatin (sodium) (cerivastatin (sodium)) Itavastatin, atorvastatin (calcium hydrate), lovastatin, pitavastatin (calcium), ZD-4522, and the like.

  Examples of the parathyroid hormone include dried thyroid, levothyroxine sodium, liothyronine sodium, propylthiouracil, thiamazole, and the like.

  Examples of growth factors include fibroblast growth factor (FGF), vascular endothelial growth factor (VEGF), hepatocyte growth factor (HGF), and insulin-like growth factor.

  Examples of the caspase-1 inhibitor include nitroflubiprofen and prarnacasan.

  Examples of the PTHrP derivative include hPTHrP and RS-66271.

  Examples of metalloproteinase inhibitors include ONO-4817.

  Examples of the farnesoid X receptor agonist include SR-45023A.

  Examples of the estrogen agonist include TSE-424, WJ-713 / MPA, lasofoxifene tartrate, estradiol, teriparatide acetate, osaterone acetate and the like.

  Examples of progesterone agonists include trimegestone.

  The above concomitant drugs are illustrative and are not limited thereto.

  Other drugs may be administered in combination of any two or more. In addition, the drugs used in combination include not only those found so far, but also those that will be found in the future based on the above-described mechanism.

[toxicity]
Since the toxicity of the 5-thia-PG compound used in the present invention is sufficiently low, the composition of the present invention, particularly a lyophilized product, can be used safely as a pharmaceutical product. In addition, since the composition of the present invention, in particular the lyophilized product, can minimize the production associated with the storage of degradation products that have not been confirmed to be toxic, the risk of exposure to the living body of 5-thia-PG compounds is reduced. Can be reduced.

  Since the pharmaceutical composition provided by the present invention, in particular the lyophilized product, is stable, it is stable and high-quality injectable preparation in both purity and form, suppressing the generation of degradation products during storage. As such, it can be provided to the clinic in a safe state without degrading the quality.

  Hereinafter, the present invention will be described in detail with reference to examples, but the present invention is not limited thereto.

  Moreover, you may change in the range which does not deviate from the range of this invention.

[Example 1]
Methyl 4-{[2-((1R, 2R, 3R) -3-hydroxy-2-{(1E, 3S) -3-hydroxy-4- [3- (methoxymethyl) phenyl] but-1-enyl} -5-Oxocyclopentyl) ethyl] sulfanyl} butanoate in aqueous solution containing α-cyclodextrin inclusion compound 1-1: maltose formulation [Formulation 1-1-1] Content: 10 μg / vial, additive: maltose Injection To the water, methyl 4-{[2-((1R, 2R, 3R) -3-hydroxy-2-{(1E, 3S) -3-hydroxy-4- [3- (methoxymethyl) phenyl] buta-1 Α-cyclodextrin inclusion compound of -enyl} -5-oxocyclopentyl) ethyl] sulfanyl} butanoate (9.7 mg: methyl 4-{[2-((1R, 2R, 3 ) -3-hydroxy-2-{(1E, 3S) -3-hydroxy-4- [3- (methoxymethyl) phenyl] but-1-enyl} -5-oxocyclopentyl) ethyl] sulfanyl} butanoate 2 mg) And crystalline maltose (10 g) were added to make 120 mL using water for injection. After preparing a uniform solution, the solution was filtered with a sterile filter (0.22 μm membrane made by Durapore) to produce the aqueous solution of the present invention. The solubility of each aqueous solution was clear and colorless. In addition, 3 lots are manufactured by this prescription, and pH after filtration of each aqueous solution is 6.00 (lot A-1), 6.00 (lot A-2), 5.58 (lot A-3). there were.

[Formulation 1-1-2] Content: 10 μg / vial, additives: maltose, citric acid Into water for injection, methyl 4-{[2-((1R, 2R, 3R) -3-hydroxy-2-{(1E , 3S) -3-hydroxy-4- [3- (methoxymethyl) phenyl] but-1-enyl} -5-oxocyclopentyl) ethyl] sulfanyl} butanoate (9.7 mg: methyl) 4-{[2-((1R, 2R, 3R) -3-hydroxy-2-{(1E, 3S) -3-hydroxy-4- [3- (methoxymethyl) phenyl] but-1-enyl}- 5-oxocyclopentyl) ethyl] sulfanyl} butanoate (2 mg) and crystalline maltose (10 g) were added, and various concentrations (0.5N, 0.05N, 0.01N) of citric acid water Adjust the pH by adding an appropriate amount of liquid, and a 120mL with water for injection. After preparing a uniform solution, the solution was filtered with a sterile filter (0.22 μm membrane made by Durapore) to produce the aqueous solution of the present invention. The solubility of each aqueous solution was clear and colorless. In addition, 6 lots are manufactured by this prescription, pH after filtration of each aqueous solution is 4.40 (lot B-1), 5.32 (lot B-2), 3.75 (lot B-3), 3.16 (Lot B-4), 3.72 (Lot B-5), 3.69 (Lot B-6).

[Formulation 1-1-3] Content: 10 μg / vial, Additive: Maltose, citric acid, sodium citrate Methyl 4-{[2-((1R, 2R, 3R) -3-hydroxy-2] Α-cyclodextrin inclusion compound of {(1E, 3S) -3-hydroxy-4- [3- (methoxymethyl) phenyl] but-1-enyl} -5-oxocyclopentyl) ethyl] sulfanyl} butanoate (9 .7 mg: methyl 4-{[2-((1R, 2R, 3R) -3-hydroxy-2-{(1E, 3S) -3-hydroxy-4- [3- (methoxymethyl) phenyl] buta-1 -Enyl} -5-oxocyclopentyl) ethyl] sulfanyl} butanoate 2 mg), crystalline maltose (10 g) and 0.01 N aqueous citric acid (10 mL). For example, to adjust the pH by adding an appropriate amount of addition 0.05N sodium citrate aqueous solution to a 120mL with water for injection. After preparing a uniform solution, the solution was filtered with a sterile filter (0.22 μm membrane made by Durapore) to produce the aqueous solution of the present invention. The solubility of each aqueous solution was clear and colorless. In addition, 3 lots are manufactured by this prescription, pH after filtration of each aqueous solution is 4.12 (lot C-1), 4.58 (lot C-2), 5.06 (lot C-3). there were.

[Formulation 1-1-4] Content: 10 μg / vial, additives: maltose, citric acid, sodium hydroxide In water for injection, methyl 4-{[2-((1R, 2R, 3R) -3-hydroxy-2 Α-cyclodextrin inclusion compound of {(1E, 3S) -3-hydroxy-4- [3- (methoxymethyl) phenyl] but-1-enyl} -5-oxocyclopentyl) ethyl] sulfanyl} butanoate (9 .7 mg: methyl 4-{[2-((1R, 2R, 3R) -3-hydroxy-2-{(1E, 3S) -3-hydroxy-4- [3- (methoxymethyl) phenyl] buta-1 -Enyl} -5-oxocyclopentyl) ethyl] sulfanyl} butanoate, 2 mg), crystalline maltose (10 g) and 0.01 N aqueous citric acid (10 mL) were added. To adjust the pH by adding an appropriate amount of addition 0.05N aqueous sodium hydroxide, it was 120mL with water for injection. After preparing a uniform solution, the solution was filtered with a sterile filter (0.22 μm membrane made by Durapore) to produce the aqueous solution of the present invention. The solubility of each aqueous solution was clear and colorless. In addition, 4 lots are manufactured by this prescription, pH after filtration of each aqueous solution is 4.11 (lot D-1), 4.58 (lot D-2), 5.09 (lot D-3), It was 5.72 (lot D-4).
1-2: Lactose formulation [Formulation 1-2-1] Content: 10 μg / vial, additive: lactose Injectable water, methyl 4-{[2-((1R, 2R, 3R) -3-hydroxy-2- Α-cyclodextrin inclusion compound of {(1E, 3S) -3-hydroxy-4- [3- (methoxymethyl) phenyl] but-1-enyl} -5-oxocyclopentyl) ethyl] sulfanyl} butanoate (9. 4 mg: methyl 4-{[2-((1R, 2R, 3R) -3-hydroxy-2-{(1E, 3S) -3-hydroxy-4- [3- (methoxymethyl) phenyl] but-1- 2 mg) and lactose (10 g) were added as enyl} -5-oxocyclopentyl) ethyl] sulfanyl} butanoate, and made up to 120 mL with water for injection. After preparing a uniform solution, the solution was filtered through a sterile filter (0.22 μm membrane made by Durapore) to produce the aqueous solution (Lot E) of the present invention. The solution of the aqueous solution was clear and colorless, and the pH was about 4 to about 5.

[Formulation 1-2-2] Content: 100 μg / vial, additive: lactose In water for injection, methyl 4-{[2-((1R, 2R, 3R) -3-hydroxy-2-{(1E, 3S) Α-cyclodextrin inclusion compound of -3-hydroxy-4- [3- (methoxymethyl) phenyl] but-1-enyl} -5-oxocyclopentyl) ethyl] sulfanyl} butanoate (94 mg: methyl 4-{[2 -((1R, 2R, 3R) -3-hydroxy-2-{(1E, 3S) -3-hydroxy-4- [3- (methoxymethyl) phenyl] but-1-enyl} -5-oxocyclopentyl) Ethyl] sulfanyl} butanoate (20 mg) and lactose (10 g) were added, and the volume was made up to 120 mL using water for injection. After preparing a uniform solution, the solution was filtered with a sterile filter (0.22 μm membrane made by Durapore) to produce the aqueous solution (Lot F) of the present invention. The solution of the aqueous solution was clear and colorless, and the pH was about 4 to about 5.

[Example 2]
0.6 mL of the aqueous solution prepared in Example 1 was filled into a silicon-coated glass vial and stored under conditions of 25 ° C. and 60% relative humidity (RH) for 24 hours. Methyl 4-{[2-((1R, 2R, 3R) -3-hydroxy-2-{(1E, 3S) -3-hydroxy-4- [3- (methoxymethyl) phenyl] in each vial after storage ] But-1-enyl} -5-oxocyclopentyl) ethyl] sulfanyl} butanoate was measured under the following conditions to evaluate the residual rate.

[Evaluation method of survival rate]
The evaluation method of the residual rate is described below, but the following notation shall conform to the general rules of the Japanese Pharmacopoeia. For example, the description “accurately” means to accurately measure up to that digit, and the description “about” means a range of ± 10%.
(1) Preparation of sample solution and standard solution <Preparation of sample solution>
For three specimens, the internal standard solution 2 described later was added to each of the samples, and methyl 4-{[2-((1R, 2R, 3R) -3-hydroxy-2-{(1E, 3S) -3-hydroxy-4] -[3- (methoxymethyl) phenyl] but-1-enyl} -5-oxocyclopentyl) ethyl] sulfanyl} butanoate and butyl 4-hydroxybenzoate were mixed so that the concentration ratio was the same as the standard solution described later . About 3 specimens, 0.4 mL was collected from each sample and mixed to prepare a sample solution.
<Preparation of standard solution>
Methyl 4-{[2-((1R, 2R, 3R) -3-hydroxy-2-{(1E, 3S) -3-hydroxy-4- [3- (methoxymethyl) phenyl] but-1-enyl} About 25 mg of -5-oxocyclopentyl) ethyl] sulfanyl} butanoate was weighed and dissolved in methanol to make exactly 100 mL. 4 mL of this liquid was accurately weighed, 4 mL of the internal standard solution 1 described later was accurately added, and methanol was further added to make exactly 100 mL. This solution was diluted with water at the time of use and used (methyl 4-{[2-((1R, 2R, 3R) -3-hydroxy-2-{(1E, 3S) -3-hydroxy-4- [3- (Methoxymethyl) phenyl] but-1-enyl} -5-oxocyclopentyl) ethyl] sulfanyl} butanoate concentration: 5 μg / mL, internal standard concentration: about 3 μg / mL).
<Preparation of internal standard solution 1>
30 mg of butyl 4-hydroxybenzoate was weighed, and methanol was added to 200 mL (butyl 4-hydroxybenzoate concentration: 150 μg / mL).
<Preparation of internal standard solution 2>
4 mL of the internal standard solution 1 was accurately weighed and methanol was added to make exactly 100 mL (butyl 4-hydroxybenzoate concentration: 6 μg / mL).
(2) High performance liquid chromatograph The sample solution was subjected to high performance liquid chromatograph under the following conditions.
<Test conditions>
Detector: ultraviolet absorptiometer (measurement wavelength: 210 nm);
Column: A stainless steel tube having an inner diameter of 4.6 mm and a length of 15 cm is filled with 5 μm of octadecylsilylated silica gel for liquid chromatography (for example, YMC-Pack ODS A302);
Column temperature: constant temperature around 50 ° C;
Mobile phase: water / acetonitrile / methanol mixed solvent (water: acetonitrile: methanol = 58: 25: 17);
Flow rate: 1.0 mL / min;
Analysis time: 40 minutes (standard solution 20 minutes);
Standard retention time: methyl 4-{[2-((1R, 2R, 3R) -3-hydroxy-2-{(1E, 3S) -3-hydroxy-4- [3- (methoxymethyl) phenyl] buta -1-enyl} -5-oxocyclopentyl) ethyl] sulfanyl} butanoate: about 15 minutes, internal standard: about 18 minutes.
(3) Calculation of residual ratio Methyl 4-{[2-((1R, 2R, 3R) -3-hydroxy-2-{(1E, 3S) -3 with respect to the peak area of the internal standard substance in the sample solution and the standard solution] It was calculated from the peak area ratio of -hydroxy-4- [3- (methoxymethyl) phenyl] but-1-enyl} -5-oxocyclopentyl) ethyl] sulfanyl} butanoate.

[result]
The obtained results are shown in Tables 1 to 4 below. In the table, the pH before storage (after filtration) and the pH after storage are described together, and the pH after storage is described as "-" for those not measured.
When crystalline maltose is used alone as an additive and the pH is in the range of about 5.5 to about 6.5, the condition of the 5-thia-PG compound is 25 ° C. and 60% relative humidity (RH). The residual rate after storage for 24 hours was about 94% to about 100% (Table 1). When a combination of crystalline maltose and citric acid is used as an additive and the pH is adjusted to about 3.5 to about 5.5, the 5-thia-PG compound at 25 ° C., 60% relative humidity (RH) The residual rate after storage for 24 hours under these conditions was about 97% to about 100% (Table 2). Further, when a combination of crystalline maltose, citric acid and sodium citrate is used as an additive and the pH is adjusted to about 4 to about 5.5, the 5-thia-PG compound at 25 ° C., 60% relative humidity (RH) The residual rate after storage for 24 hours under the conditions of) was about 99% to about 100% (Table 3). Furthermore, when a combination of crystalline maltose, citric acid and sodium hydroxide is used as an additive, and the pH is adjusted to about 4 to about 6, the 5-thia-PG compound at 25 ° C., 60% relative humidity (RH) The residual rate after storage for 24 hours under these conditions was about 98% to about 100% (Table 4).

[Example 3]
Methyl 4-{[2-((1R, 2R, 3R) -3-hydroxy-2-{(1E, 3S) -3-hydroxy-4- [3- (methoxymethyl) phenyl] but-1-enyl} -5-Oxocyclopentyl) ethyl] sulfanyl} butanoate Preparation of Lyophilized Product Containing α-Cyclodextrin Inclusion Compound 3-1: Maltose Formulation [Formulation 3-1-1] Content: 10 μg / vial, Additives: Maltose In water for injection, methyl 4-{[2-((1R, 2R, 3R) -3-hydroxy-2-{(1E, 3S) -3-hydroxy-4- [3- (methoxymethyl) phenyl] butane -1-enyl} -5-oxocyclopentyl) ethyl] sulfanyl} butanoate inclusion compound of α-cyclodextrin (9.7 mg: methyl 4-{[2-((1R, 2R 3 mg of 3R) -3-hydroxy-2-{(1E, 3S) -3-hydroxy-4- [3- (methoxymethyl) phenyl] but-1-enyl} -5-oxocyclopentyl) ethyl] sulfanyl} butanoate ) And crystalline maltose (10 g) were added to make 120 mL using water for injection. After preparing a uniform solution (pH 5.66), the solution was filtered through a sterile filter (Durapore 0.22 μm membrane), and each 0.6 mL was filled into a silicon-coated glass vial. The vial was cooled to −40 ° C. or lower to freeze the solution, then heated gradually at a vacuum of about 20 Pascal and dried at 10 ° C. for 7 hours. Then, the freeze-dried product (lot a-1) of the present invention was produced by final drying at 45 ° C. The appearance of the lyophilized product was a white mass or powder.

[Formulation 3-1-2] Content: 10 μg / vial, Additives: Maltose, citric acid To water for injection, methyl 4-{[2-((1R, 2R, 3R) -3-hydroxy-2-{(1E , 3S) -3-hydroxy-4- [3- (methoxymethyl) phenyl] but-1-enyl} -5-oxocyclopentyl) ethyl] sulfanyl} butanoate (9.7 mg: methyl) 4-{[2-((1R, 2R, 3R) -3-hydroxy-2-{(1E, 3S) -3-hydroxy-4- [3- (methoxymethyl) phenyl] but-1-enyl}- 5 mg as 5-oxocyclopentyl) ethyl] sulfanyl} butanoate), crystalline maltose (10 g) and 0.01 N aqueous citric acid (10 mL) to adjust pH (p 3.64), and was 120mL with water for injection. After preparing a uniform solution, the solution was filtered through a sterile filter (Durapore 0.22 μm membrane), and each 0.6 mL was filled into a silicon-coated glass vial. The vial was cooled to −40 ° C. or lower to freeze the solution, then heated gradually at a vacuum of about 20 Pascal and dried at 10 ° C. for 7 hours. Then, the freeze-dried product (lot b-1) of the present invention was produced by final drying at 45 ° C. The appearance of the lyophilized product was a white mass or powder.

[Formulation 3-1-3] Content: 10 μg / vial, additive: maltose, citric acid, sodium citrate In water for injection, methyl 4-{[2-((1R, 2R, 3R) -3-hydroxy-2 Α-cyclodextrin inclusion compound of {(1E, 3S) -3-hydroxy-4- [3- (methoxymethyl) phenyl] but-1-enyl} -5-oxocyclopentyl) ethyl] sulfanyl} butanoate (9 .7 mg: methyl 4-{[2-((1R, 2R, 3R) -3-hydroxy-2-{(1E, 3S) -3-hydroxy-4- [3- (methoxymethyl) phenyl] buta-1 -Enyl} -5-oxocyclopentyl) ethyl] sulfanyl} butanoate 2 mg), crystalline maltose (10 g) and 0.01 N aqueous citric acid (10 mL). For example, further adjust the pH by adding an appropriate amount of 0.05N aqueous sodium citrate (pH4.07, pH4.53, pH5.09) and to obtain a 120mL with water for injection. After preparing a uniform solution, the solution was filtered through a sterile filter (Durapore 0.22 μm membrane), and each 0.6 mL was filled into a silicon-coated glass vial. The vial was cooled to −40 ° C. or lower to freeze the solution, then heated gradually at a vacuum of about 20 Pascal and dried at 10 ° C. for 7 hours. Thereafter, the lyophilized product of the present invention is subjected to final drying at 45 ° C. (Lot is distinguished by pH before filtration: lot c-1 (pH 4.07), lot c-2 (pH 4.53), lot c-3 (PH 5.09)). The appearance of the lyophilized product was a white mass or powder.

[Formulation 3-1-4] Content: 10 μg / vial, additive: maltose, citric acid, sodium hydroxide In water for injection, methyl 4-{[2-((1R, 2R, 3R) -3-hydroxy-2 Α-cyclodextrin inclusion compound of {(1E, 3S) -3-hydroxy-4- [3- (methoxymethyl) phenyl] but-1-enyl} -5-oxocyclopentyl) ethyl] sulfanyl} butanoate (9 .7 mg: methyl 4-{[2-((1R, 2R, 3R) -3-hydroxy-2-{(1E, 3S) -3-hydroxy-4- [3- (methoxymethyl) phenyl] buta-1 -Enyl} -5-oxocyclopentyl) ethyl] sulfanyl} butanoate, 2 mg), crystalline maltose (10 g) and 0.01 N aqueous citric acid (10 mL) were added. Further added suitable amount of 0.05N sodium hydroxide aqueous solution adjusted pH (pH4.07, pH4.54, pH5.04) and to obtain a 120mL with water for injection. After preparing a uniform solution, the solution was filtered through a sterile filter (Durapore 0.22 μm membrane), and each 0.6 mL was filled into a silicon-coated glass vial. The vial was cooled to −40 ° C. or lower to freeze the solution, then heated gradually at a vacuum of about 20 Pascal and dried at 10 ° C. for 7 hours. Thereafter, the lyophilized product of the present invention is subjected to final drying at 45 ° C. (Lot is distinguished by pH before filtration: lot d-1 (pH 4.07), lot d-2 (pH 4.54), lot d-3) (PH 5.04)). The appearance of the lyophilized product was a white mass or powder.

[Example 4]
The freeze-dried product produced in Example 3 was (1) 1 week at 60 ° C, (2) 2 weeks at 60 ° C, (3) 3 weeks at 60 ° C, (4) It was stored for 1 month at 40 ° C. and 75% relative humidity (RH). (A) Appearance, (b) Appearance at re-dissolution, (c) pH at re-dissolution, (d) Water content, and (e) Residual rate for each sample after storage and before storage (Initial: initial) Each item of was evaluated using the following evaluation methods.
[Evaluation methods]
(A) The appearance of the freeze-dried product before and after storage of the appearance was observed.
(B) Appearance at the time of redissolving The lyophilized product before and after storage was dissolved in 0.6 mL of water for injection and the appearance was observed.
(C) pH during re-dissolution
The lyophilized product before and after storage was dissolved in 0.6 mL of water for injection, and the pH was measured.
(D) Water content The water content in the lyophilized product before storage was measured by the Karl Fischer method (volume method) using the following test conditions.
<Test conditions>
Apparatus: Moisture analyzer (Mitsubishi Chemical) CA-06
Method: Capacity method (KF)
Reagents: Dehydrated solvent (GEX), Karl Fischer reagent (SS-X) 3 mg (Mitsubishi Chemical)
(E) Residual ratio Before and after storage, freeze-dried products were used as samples, and methyl 4-{[2-((1R, 2R, 3R) -3-hydroxy-2- { The residual rate of (1E, 3S) -3-hydroxy-4- [3- (methoxymethyl) phenyl] but-1-enyl} -5-oxocyclopentyl) ethyl] sulfanyl} butanoate was measured. In addition, as for the sample solution, an appropriate amount of water and an internal standard solution 2 described later were added to each of three specimens, and methyl 4-{[2-((1R, 2R, 3R) -3-hydroxy-2- { The concentration ratio of (1E, 3S) -3-hydroxy-4- [3- (methoxymethyl) phenyl] but-1-enyl} -5-oxocyclopentyl) ethyl] sulfanyl} butanoate and butyl 4-hydroxybenzoate is It mixed so that it might become the same as the standard solution of, and it prepared by collecting 0.4 mL each from these 3 samples, and mixing.
[result]
The results obtained are shown in Tables 5 to 9 below (Table 5: Initial; Table 6: 60 ° C., 1 week; Table 7: 60 ° C., 2 weeks; Table 8: 60 ° C., 4 weeks; Table 9: 40 ° C.) , 75% RH, 1 month). When a freeze-dried product is produced using an aqueous solution containing crystalline maltose alone as an additive (when no pH regulator is added to the aqueous solution: lot a-1), after storage at 60 ° C. for 2 weeks The residual ratio of 5-thia-PG compound in the lyophilized product was about 85% to about 100% (Table 7). When a freeze-dried product is produced using an aqueous solution in which crystalline maltose and citric acid are combined as an additive and the pH is adjusted to about 3.5 to about 4 (lot b-1), The residual ratio of 5-thia-PG compound in the lyophilized product after storage for 2 weeks was about 90% to about 100% (Table 7). Further, when freeze-dried products are produced using an aqueous solution in which crystalline maltose, citric acid and sodium citrate are combined as additives and the pH is adjusted to about 4 to about 5.5 (Lot c-1, Lot c-2) In lot c-3), the residual ratio of 5-thia-PG compound in the lyophilized product after storage for 2 weeks at 60 ° C. was about 92% to about 100% (Table 7). Furthermore, when freeze-dried products are produced using an aqueous solution in which crystalline maltose, citric acid and sodium hydroxide are combined as additives and the pH is adjusted to about 4 to about 5.5 (lot d-1, lot d- 2, Lot d-3), the residual ratio of 5-thia-PG compound in the lyophilized product after storage for 2 weeks at 60 ° C. was about 93% to about 100% (Table 7). . Regarding the appearance and appearance upon re-dissolution, no change due to storage was observed in any specimen. Among the lyophilized products studied, the lyophilized product (lot lot) in which the residual ratio of 5-thia-PG compound after storage for 2 weeks at 60 ° C. was particularly good (the residual ratio of about 95% to about 100%) c-1, lot d-1, and lot d-2) are stable even when stored for a longer period of time. For example, a residual ratio of about 92% to about 100% after storage for 4 weeks at 60 ° C. (Table 8). Moreover, in the residual ratio after storage for 1 month under the conditions of 40 ° C. and 75% RH, all the freeze-dried products showed good results. In particular, crystalline maltose, citric acid and sodium citrate were used as additives. In combination, freeze-dried products (lot c-1, lot c-2, lot c-3) produced using an aqueous solution with a pH adjusted to about 4 to about 5.5, crystalline maltose, citric acid and Good results were obtained with a freeze-dried product (lot d-1) produced using an aqueous solution in which sodium hydroxide was combined and the pH was adjusted to about 4 (Table 9).

[Example 5]
[Formulation 5-1-1] Content: 500 μg / vial, additive: maltose or lactose In water for injection, methyl 4-{[2-((1R, 2R, 3R) -3-hydroxy-2-{(1E, 3S) -3-hydroxy-4- [3- (methoxymethyl) phenyl] but-1-enyl} -5-oxocyclopentyl) ethyl] sulfanyl} butanoate [alpha] -cyclodextrin inclusion compound (480 mg: methyl 4- { [2-((1R, 2R, 3R) -3-hydroxy-2-{(1E, 3S) -3-hydroxy-4- [3- (methoxymethyl) phenyl] but-1-enyl} -5-oxo Cyclopentyl) ethyl] sulfanyl} butanoate, 100 mg), crystalline maltose (10 g) or lactose (10 g), and 120 mL with water for injection It was. After preparing a uniform solution, the solution was filtered through a sterile filter (Durapore 0.22 μm membrane), and each 0.6 mL was filled into a silicon-coated glass vial. The vial was cooled to −40 ° C. or lower to freeze the solution, then heated gradually at a vacuum of about 20 Pascal and dried at 10 ° C. for 7 hours. Then, the freeze-dried product was manufactured by final drying at 45 degreeC. The appearance of the freeze-dried product was white and cracked in the cake. When maltose was used alone as an additive, the pH of the aqueous solution before filtration showed a value around 7.

[Example 6]
The freeze-dried product produced in Example 5 was stored (1) for 1 week at 60 ° C., (2) for 2 weeks at 60 ° C., and (3) for 3 weeks at 60 ° C. (A) Appearance, (b) Appearance at re-dissolution, (c) pH at re-dissolution, (d) Water content, and (e) Residual rate for each sample after storage and before storage (Initial: initial) Each item was evaluated using the evaluation method described in Example 4.
[result]
The obtained results are shown in Tables 10 to 13 below (Table 10: Initial; Table 11: 60 ° C., 1 week; Table 12: 60 ° C., 2 weeks; Table 13: 60 ° C., 4 weeks). Regarding the appearance and the appearance upon re-dissolution, no change due to storage was observed in any specimen. As for the residual rate, the results with a pH of 6.5 or less (pH during re-dissolution), that is, those using lactose showed better results.

[Example 7]
7-1: Lactose formulation [Prescription 7-1-1] Content: 10 μg / vial, additive: lactose Injectable water, methyl 4-{[2-((1R, 2R, 3R) -3-hydroxy-2- Α-cyclodextrin inclusion compound of {(1E, 3S) -3-hydroxy-4- [3- (methoxymethyl) phenyl] but-1-enyl} -5-oxocyclopentyl) ethyl] sulfanyl} butanoate (9. 4 mg: methyl 4-{[2-((1R, 2R, 3R) -3-hydroxy-2-{(1E, 3S) -3-hydroxy-4- [3- (methoxymethyl) phenyl] but-1- 2 mg) and lactose (10 g) were added as enyl} -5-oxocyclopentyl) ethyl] sulfanyl} butanoate, and made up to 120 mL with water for injection. After preparing a uniform solution, the solution was filtered through a sterile filter (Durapore 0.22 μm membrane), and each 0.6 mL was filled into a silicon-coated glass vial. The vial was cooled to −40 ° C. or lower to freeze the solution, then heated gradually at a vacuum of about 20 Pascal and dried at 10 ° C. for 7 hours. Then, the freeze-dried product of the present invention was produced by final drying at 45 ° C. The appearance of the lyophilized product was a white mass or powder, and the water content was 1%.

[Formulation 7-1-2] Content: 100 μg / vial, additive: lactose In water for injection, methyl 4-{[2-((1R, 2R, 3R) -3-hydroxy-2-{(1E, 3S) Α-cyclodextrin inclusion compound of -3-hydroxy-4- [3- (methoxymethyl) phenyl] but-1-enyl} -5-oxocyclopentyl) ethyl] sulfanyl} butanoate (94 mg: methyl 4-{[2 -((1R, 2R, 3R) -3-hydroxy-2-{(1E, 3S) -3-hydroxy-4- [3- (methoxymethyl) phenyl] but-1-enyl} -5-oxocyclopentyl) Ethyl] sulfanyl} butanoate (20 mg) and lactose (10 g) were added, and the volume was made up to 120 mL using water for injection. After preparing a uniform solution, the solution was filtered through a sterile filter (Durapore 0.22 μm membrane), and each 0.6 mL was filled into a silicon-coated glass vial. The vial was cooled to −40 ° C. or lower to freeze the solution, then heated gradually at a vacuum of about 20 Pascal and dried at 10 ° C. for 7 hours. Then, the freeze-dried product of the present invention was produced by final drying at 45 ° C. The appearance of the lyophilized product was a white mass or powder, and the water content was 1%.
7-2: Maltose formulation [Formulation 7-2-1] Content: 10 μg / vial, additives: maltose, citric acid, sodium hydroxide Injectable water, methyl 4-{[2-((1R, 2R, 3R) Α-cyclo of -3-hydroxy-2-{(1E, 3S) -3-hydroxy-4- [3- (methoxymethyl) phenyl] but-1-enyl} -5-oxocyclopentyl) ethyl] sulfanyl} butanoate Dextrin inclusion compound (9.7 mg: methyl 4-{[2-((1R, 2R, 3R) -3-hydroxy-2-{(1E, 3S) -3-hydroxy-4- [3- (methoxymethyl ) Phenyl] but-1-enyl} -5-oxocyclopentyl) ethyl] sulfanyl} butanoate 2 mg) and crystalline maltose (10 g) are added, The pH was adjusted to about 4.0 by adding appropriate amounts of an aqueous solution with an aqueous solution of sodium hydroxide, was 120mL with water for injection. After preparing a uniform solution, the solution was filtered through a sterile filter (Durapore 0.22 μm membrane), and each 0.6 mL was filled into a silicon-coated glass vial. The vial was cooled to −40 ° C. or lower to freeze the solution, then heated gradually at a vacuum of about 20 Pascal and dried at 10 ° C. for 7 hours. Then, the freeze-dried product of the present invention was produced by final drying at 45 ° C. The appearance of the lyophilized product was a white mass or powder, and the water content was 0.7%.

The composition of the present invention, particularly a lyophilized product, is methyl 4-{[2-((1R, 2R, 3R) useful for the prevention, treatment and / or progression inhibition of bone loss diseases such as vertebral fractures. ) -3-hydroxy-2-{(1E, 3S) -3-hydroxy-4- [3- (methoxymethyl) phenyl] but-1-enyl} -5-oxocyclopentyl) ethyl] sulfanyl} butanoate It can be provided to the clinic in a safe state without deteriorating the quality while maintaining a high residual rate of ingredients, and its applicability as a pharmaceutical is extremely high.

Claims (30)

  (1) Methyl 4-{[2-((1R, 2R, 3R) -3-hydroxy-2-{(1E, 3S) -3-hydroxy-4- [3- (methoxymethyl) phenyl] buta-1 -Enyl} -5-oxocyclopentyl) ethyl] sulfanyl} butanoate, a solvate thereof, or an inclusion compound thereof, and (2) a saccharide, and optionally (3) a pH regulator A stable pharmaceutical composition.   The composition according to claim 1, which is a freeze-dried product.   The composition according to claim 1, wherein the inclusion compound is an α-cyclodextrin inclusion compound.   The composition according to claim 1, wherein the saccharide is lactose or maltose.   The composition according to claim 4, wherein the saccharide is maltose having a pH of 4.5 to 6.5.   Methyl 4-{[2-((1R, 2R, 3R) -3-hydroxy-2-{(1E, 3S) -3-hydroxy-4- [3- (methoxymethyl) phenyl] but-1-enyl} The composition according to claim 1, comprising 500 parts by weight to 50,000 parts by weight of a saccharide with respect to 1 part by weight of -5-oxocyclopentyl) ethyl] sulfanyl} butanoate.   The composition according to claim 1, wherein the pH regulator is one or more selected from organic acids, metal hydroxides, and organic acid metal salts.   The composition according to claim 7, wherein the pH adjusting agent is (1) citric acid and sodium hydroxide, or (2) citric acid and sodium citrate.   (1) Methyl 4-{[2-((1R, 2R, 3R) -3-hydroxy-2-{(1E, 3S) -3-hydroxy-4- [3- (methoxymethyl) phenyl] buta-1 -Enyl} -5-oxocyclopentyl) ethyl] sulfanyl} butanoate inclusion compound and (2) maltose at pH 4.5 to 6.5, and (3) citric acid and sodium hydroxide A stable pharmaceutical composition comprising:   The composition according to claim 9, which is a freeze-dried product.   Methyl 4-{[2-((1R, 2R, 3R) -3-hydroxy-2-{(1E, 3S) -3-hydroxy-4- [3- (methoxymethyl) phenyl] but-1-enyl} The composition according to claim 9, comprising 1000 parts by weight to 10,000 parts by weight of maltose having a pH of 4.5 to 6.5 per 1 part by weight of -5-oxocyclopentyl) ethyl] sulfanyl} butanoate.   Methyl 4-{[2-((1R, 2R, 3R) -3-hydroxy-2-{(1E, 3S) -3-hydroxy-4- [3- (methoxymethyl) phenyl] but-1-enyl} The composition according to claim 11, comprising 5000 parts by weight of maltose having a pH of 4.5 to 6.5 per 1 part by weight of -5-oxocyclopentyl) ethyl] sulfanyl} butanoate.   The composition according to claim 2 or 10, which is substantially free of water or has a water content of 1% or less.   After storage at 60 ° C. for 2 weeks, methyl 4-{[2-((1R, 2R, 3R) -3-hydroxy-2-{(1E, 3S) -3-hydroxy-4- [3- (methoxymethyl) The stable composition according to claim 2 or 10, wherein the residual ratio of) phenyl] but-1-enyl} -5-oxocyclopentyl) ethyl] sulfanyl} butanoate is 85% or more.   The stable composition according to claim 14, wherein the residual ratio is 95% or more.   11. A stable composition according to claim 2 or 10 having a form substantially identical to the initial form after storage at 60 ° C. for 2 weeks.   The composition according to claim 1, 2, 9, or 10, which is an injectable preparation.   The composition according to claim 1, 2, 9, or 10, which is a preventive, therapeutic and / or progression inhibitor for EP4-mediated diseases.   The composition according to claim 18, wherein the EP4-mediated disease is a bone disease.   The composition according to claim 19, wherein the bone disease is a vertebral fracture.   An injection container filled with the composition according to claim 2 or 10.   The container according to claim 21, which is a vial.   1 unit to 12 μg of methyl 4-{[2-((1R, 2R, 3R) -3-hydroxy-2-{(1E, 3S) -3-hydroxy-4- [3- (methoxymethyl) per unit form The container according to claim 21, which comprises :)) phenyl] but-1-enyl} -5-oxocyclopentyl) ethyl] sulfanyl} butanoate.   2 μg, 5 μg, 6 μg or 10 μg of methyl 4-{[2-((1R, 2R, 3R) -3-hydroxy-2-{(1E, 3S) -3-hydroxy-4- [3 per unit form] The container according to claim 23, comprising-(methoxymethyl) phenyl] but-1-enyl} -5-oxocyclopentyl) ethyl] sulfanyl} butanoate.   (1) Methyl 4-{[2-((1R, 2R, 3R) -3-hydroxy-2-{(1E, 3S) -3-hydroxy-4- [3- (methoxymethyl) phenyl] buta-1 -Enyl} -5-oxocyclopentyl) ethyl] sulfanyl} butanoate, a solvate thereof, or an inclusion compound thereof, and (2) a saccharide, and optionally (3) a pH regulator A method for producing a stable freeze-dried product, comprising subjecting an aqueous solution having a pH of 3 to 6.5 to freeze-drying.   (1) Methyl 4-{[2-((1R, 2R, 3R) -3-hydroxy-2-{(1E, 3S) -3-hydroxy-4- [3- (methoxymethyl) phenyl] buta-1 -Enyl} -5-oxocyclopentyl) ethyl] sulfanyl} butanoate, a solvate thereof, or an inclusion compound thereof, and (2) a saccharide, and optionally (3) a pH regulator Methyl 4-{[2-((1R, 2R, 3R) -3-hydroxy- in a lyophilized product obtained by freeze-drying an aqueous solution having a pH of 3 to 6.5 to obtain a lyophilized product] A method for stabilizing 2-{(1E, 3S) -3-hydroxy-4- [3- (methoxymethyl) phenyl] but-1-enyl} -5-oxocyclopentyl) ethyl] sulfanyl} butanoate.   A method for preventing, treating, and / or suppressing progression of an EP4-mediated disease in a mammal, comprising administering an effective amount of the composition according to claim 1, 2, 9, or 10 to the mammal.   Use of the composition according to claim 1, 2, 9 or 10 for producing a preventive, therapeutic and / or progression inhibitor for EP4-mediated diseases.   Per unit form, (1) methyl 4-{[2-((1R, 2R, 3R) -3-hydroxy-2-{(1E, 3S) -3-hydroxy-4- [3- (methoxymethyl)] Phenyl] but-1-enyl} -5-oxocyclopentyl) ethyl] sulfanyl} butanoate containing 1 μg to 20 μg of methyl 4-{[2-((1R, 2R, 3R) -3-hydroxy-2-{(1E , 3S) -3-hydroxy-4- [3- (methoxymethyl) phenyl] but-1-enyl} -5-oxocyclopentyl) ethyl] sulfanyl} butanoate, and (2) methyl 4-{[2-((1R, 2R, 3R) -3-hydroxy-2-{(1E, 3S) -3-hydroxy-4- [3- (methoxymethyl) phenyl] Tert-1-enyl} -5-oxocyclopentyl) ethyl] sulfanyl} butanoate with a stable lyophilized product containing 1000 to 10,000 parts by weight of lactose and 1% or less water content. Vials. Per unit form, (1) methyl 4-{[2-((1R, 2R, 3R) -3-hydroxy-2-{(1E, 3S) -3-hydroxy-4- [3- (methoxymethyl)] Methyl 4-{[2-((1R, 2R, 3R) -3-hydroxy-2-{(1E, 3S) containing 10 μg as phenyl] but-1-enyl} -5-oxocyclopentyl) ethyl] sulfanyl} butanoate ) -3-hydroxy-4- [3- (methoxymethyl) phenyl] but-1-enyl} -5-oxocyclopentyl) ethyl] sulfanyl} butanoate, and (2) methyl 4- {[2-((1R, 2R, 3R) -3-hydroxy-2-{(1E, 3S) -3-hydroxy-4- [3- (methoxymethyl) phenyl] but-1-ene Nil} -5-oxocyclopentyl) ethyl] sulfanyl} butanoate A vial filled with a stable lyophilized product containing 5000 parts by weight of lactose and having a water content of 1% or less.