Carrier composition

Abstract

The present invention provides a carrier composition that is less likely to cause filter clogging during sterilizing by filtration in formulation. The carrier composition according to the present invention is used in a pharmaceutical preparation produced by mixing with an active ingredient in a form dissolved in a solvent and then sterilizing by filtration. The carrier composition comprises: a hyaluronic acid derivative component including hyaluronic acid and a hyaluronic acid derivative having a steryl group introduced therein; and a cryoprotectant. The hyaluronic acid derivative has a repeating unit represented by general formula (I). The ratio of the introduced steryl group to disaccharide repeating units derived from the hyaluronic acid and the hyaluronic acid derivative is 25-60%. The carrier composition has a cryoprotectant content of 6-1750 parts by mass relative to 10 parts by mass of the hyaluronic acid derivative component and is a freeze-dried product.

Description

Carrier Composition

[0001] The present invention relates to a carrier composition. This application claims priority to Japanese Patent Application Nos. 2024-064159 and 2024-064002, filed on April 11, 2024, the contents of which are incorporated herein by reference.

[0002] In recent years, biopharmaceuticals, which are pharmaceuticals containing proteins, peptides, or nucleic acids as active ingredients, have been put into practical use, and their number continues to increase year by year. Biopharmaceuticals can fulfill unmet medical needs that traditional small molecule drugs could not. However, they have issues such as poor absorption from the digestive tract or mucous membranes, instability in the body, and a short half-life in the blood. As a result, biopharmaceuticals require frequent injections, which places a significant burden on both patients and medical professionals. Therefore, there is a need for sustained-release drug carriers, i.e., carriers for sustained-release drug delivery systems, that can encapsulate biopharmaceuticals without impairing their pharmacological activity and gradually release the active ingredient in the body.

[0003] Under these circumstances, Patent Document 1 proposes a carrier for sustained-release drug delivery systems, which is made of a hyaluronic acid derivative with excellent safety. This hyaluronic acid derivative spontaneously aggregates in aqueous solution, and can efficiently encapsulate drugs, especially biopharmaceuticals, while maintaining their biological activity. It aggregates under physiological salt concentration, or disperses under physiological salt concentration, and has good blood retention. This hyaluronic acid derivative can be used as a carrier that can efficiently encapsulate many drugs while maintaining pharmacological activity, and as a blood sustained-release carrier and targeting carrier with excellent blood retention, especially when biopharmaceuticals are used as active ingredients. Furthermore, it is said that it can also be used as a sustained-release carrier for local areas, such as subcutaneous, that can sustainably release drugs.

[0004] International Publication No. 2010 / 053140

[0005] The carrier for sustained-release drug delivery systems made of hyaluronic acid derivatives described in Patent Document 1 has the problem that during formulation, the carrier is dissolved in a solvent, and then mixed with an active ingredient, and the resulting mixture is easily clogged in the filter during the sterilization filtration process.

[0006] The present invention has been made in view of the above circumstances, and provides a carrier composition that can suppress filter clogging during sterile filtration in formulation.

[0007] That is, the present invention includes the following aspects: (1) A carrier composition used in a formulation obtained by mixing an active ingredient in a form dissolved in a solvent and then sterilizing and filtering the resulting carrier composition, the carrier composition comprising: a hyaluronic acid derivative component containing hyaluronic acid and a hyaluronic acid derivative having a steryl group introduced therein; and a cryoprotectant, the hyaluronic acid derivative having a repeating unit represented by the following general formula (I), the introduction rate of the steryl group relative to the repeating units of disaccharides derived from the hyaluronic acid and the hyaluronic acid derivative being 25% or more and 60% or less, the content of the cryoprotectant being 6 parts by mass or more and 1750 parts by mass or less relative to 10 parts by mass of the hyaluronic acid derivative component, and the carrier composition being a lyophilized product.

[0008]

[0009] (In the formula, R １ , R ２ , R ３ , and R ４ are each independently a hydrogen atom, C １－６ Alkyl, formyl and C １－６ alkylcarbonyl; Z represents a direct bond or a peptide linker consisting of any amino acid residues of 2 to 30; X １ is represented by the following formula: ｂ -R, -NR ｂ -COO-R, -NR ｂ -CO-R, -NR ｂ -CO-NR ｃ -R, -COO-R, -O-COO-R, -SR, -CO-Y ａ-SR, -O-CO-Y ｂ -SR, -NR ｂ -CO-Y ｂ -SR, and -S-SR, a group selected from the group consisting of groups represented by R ａ , R ｂ and R ｃ are each independently a hydrogen atom, C １－２０ Alkyl, Amino C ２－２０ Alkyl and hydroxy C ２－２０ alkyl, where the alkyl portion of the group is selected from the group consisting of —O— and —NR ｆ - may be inserted with a group selected from the group consisting of; R ｆ is a hydrogen atom, C １－１２ Alkyl, Amino C ２－１２ Alkyl and hydroxy C ２－１２ alkyl, the alkyl portion of which may be interrupted by a group selected from the group consisting of -O- and -NH-; R is a steryl group; Y is C ２－３０ Alkylene, or -(CH ２ CH ２ O) ｍ -CH ２ CH ２ -, where the alkylene is -O-, -NR ｇ - and -S-S- may be inserted; R ｇ is a hydrogen atom, C １－２０ Alkyl, Amino C ２－２０ Alkyl and hydroxy C ２－２０ alkyl, the alkyl portion of which may be inserted with a group selected from the group consisting of —O— and —NH—; Y ａ is C １－５ is alkylene; Y ｂ is C ２－８ Alkylene or C ２－８ alkenylene; and m is an integer of 1 or more and 100 or less.

[0010] (2) The carrier composition according to (1), wherein the content of the cryoprotectant is 6 parts by mass or more and 80 parts by mass or less relative to 10 parts by mass of the hyaluronic acid derivative component. (3) The molecular weight of the cryoprotectant is 200 or more. The carrier composition according to (1) or (2). (4) The carrier composition according to (1) or (2), wherein the cryoprotectant is a sugar or a sugar alcohol. (5) The carrier composition according to (4), wherein the sugar is one or more selected from the group consisting of sucrose, trehalose, and glucose. (6) The carrier composition according to (4) or (5), wherein the sugar alcohol is sorbitol. (7) The carrier composition according to any one of (1) to (6), wherein R is a cholesteryl group or a phytosteryl group. (8) The carrier composition according to any one of (1) to (7), wherein R is a cholesteryl group. (9) The carrier composition according to any one of (1) to (8), wherein the molecular weight of the hyaluronic acid derivative component is 1,000 or more and less than 20,000. (10) The carrier composition according to any one of (1) to (9), wherein, after the carrier composition is pulverized in a mortar, 30% by mass or more of the fraction passing through a sieve with a mesh size of 2.0 mm is measured by the dry sieving test method specified in JIS K 0069:1992. (11) A carrier composition used in a formulation obtained by mixing an active ingredient in a form dissolved in a solvent and then sterilizing and filtering the mixture, the carrier composition comprising: a hyaluronic acid derivative component containing hyaluronic acid and a hyaluronic acid derivative having a steryl group introduced therein; and a cryoprotectant; wherein the hyaluronic acid derivative has a repeating unit represented by the following general formula (I); the introduction rate of the steryl group relative to the repeating units of disaccharides derived from the hyaluronic acid and the hyaluronic acid derivative is 0.1% or more and less than 25%; the content of the cryoprotectant is 28 parts by mass or more and 1750 parts by mass or less per 10 parts by mass of the hyaluronic acid derivative component; and the salt content is 0.001 parts by mass or more and less than 0.500 parts by mass per 10 parts by mass of the hyaluronic acid derivative component; and the carrier composition is a freeze-dried product. (In the formula, R １ , R ２ , R ３ , and R ４ are each independently a hydrogen atom, C １－６Alkyl, formyl and C １－６ alkylcarbonyl; Z represents a direct bond or a peptide linker consisting of any amino acid residues of 2 to 30; X １ is represented by the following formula: ｂ -R, -NR ｂ -COO-R, -NR ｂ -CO-R, -NR ｂ -CO-NR ｃ -R, -COO-R, -O-COO-R, -SR, -CO-Y ａ -SR, -O-CO-Y ｂ -SR, -NR ｂ -CO-Y ｂ -SR, and -S-SR, a group selected from the group consisting of groups represented by R ａ , R ｂ and R ｃ are each independently a hydrogen atom, C １－２０ Alkyl, Amino C ２－２０ Alkyl and hydroxy C ２－２０ alkyl, where the alkyl portion of the group is selected from the group consisting of —O— and —NR ｆ - may be inserted with a group selected from the group consisting of; R ｆ is a hydrogen atom, C １－１２ Alkyl, Amino C ２－１２ Alkyl and hydroxy C ２－１２ alkyl, the alkyl portion of which may be interrupted by a group selected from the group consisting of -O- and -NH-; R is a steryl group; Y is C ２－３０ Alkylene, or -(CH ２ CH ２ O) ｍ -CH ２ CH ２ -, where the alkylene is -O-, -NR ｇ - and -S-S- may be inserted; R ｇ is a hydrogen atom, C １－２０ Alkyl, Amino C ２－２０ Alkyl and hydroxy C ２－２０alkyl, the alkyl portion of which may be inserted with a group selected from the group consisting of —O— and —NH—; Y ａ is C １－５ is alkylene; Y ｂ is C ２－８ Alkylene or C ２－８ (12) The carrier composition according to (11), wherein the content of the cryoprotectant is 28 parts by mass or more and 185 parts by mass or less relative to 10 parts by mass of the hyaluronic acid derivative component. (13) The molecular weight of the cryoprotectant is 200 or more. (14) The carrier composition according to (11) or (12), wherein the cryoprotectant is a sugar or a sugar alcohol. (15) The carrier composition according to (14), wherein the sugar is sucrose or maltose. (16) The carrier composition according to (14), wherein the sugar alcohol is sorbitol. (17) The carrier composition according to any one of (11) to (16), wherein R is a cholesteryl group or a phytosteryl group. (18) The carrier composition according to any one of (11) to (17), wherein R is a cholesteryl group. (19) The carrier composition according to any one of (11) to (18), wherein the molecular weight of the hyaluronic acid derivative component is 1,000 or more but less than 1,000,000. (20) The carrier composition according to any one of (11) to (19), wherein, after the carrier composition is pulverized in a mortar, 30% by mass or more of the particles pass through a 2.0 mm sieve according to the dry sieving test method specified in JIS K 0069: 1992. (21) A method for producing a carrier composition, the method comprising: a step (A) of adjusting the concentration of the hyaluronic acid derivative component in an aqueous solution to 4 mg / mL or more and 100 mg / mL or less; a step (B) of adding a cryoprotectant to the aqueous solution after the step (A); and a step (C) of freeze-drying the aqueous solution containing the hyaluronic acid derivative component and the cryoprotectant. (22) The method according to (21), wherein the hyaluronic acid derivative has a repeating unit represented by the following general formula (I): (In the formula, R １ , R ２ , R３ , and R ４ are each independently a hydrogen atom, C １－６ Alkyl, formyl and C １－６ alkylcarbonyl; Z represents a direct bond or a peptide linker consisting of any amino acid residues of 2 to 30; X １ is represented by the following formula: ｂ -R, -NR ｂ -COO-R, -NR ｂ -CO-R, -NR ｂ -CO-NR ｃ -R, -COO-R, -O-COO-R, -SR, -CO-Y ａ -SR, -O-CO-Y ｂ -SR, -NR ｂ -CO-Y ｂ -SR, and -S-SR, a group selected from the group consisting of groups represented by R ａ , R ｂ and R ｃ are each independently a hydrogen atom, C １－２０ Alkyl, Amino C ２－２０ Alkyl and hydroxy C ２－２０ alkyl, where the alkyl portion of the group is selected from the group consisting of —O— and —NR ｆ - may be inserted with a group selected from the group consisting of; R ｆ is a hydrogen atom, C １－１２ Alkyl, Amino C ２－１２ Alkyl and hydroxy C ２－１２ alkyl, the alkyl portion of which may be interrupted by a group selected from the group consisting of -O- and -NH-; R is a steryl group; Y is C ２－３０ Alkylene, or -(CH ２ CH ２ O) ｍ -CH ２ CH ２ -, where the alkylene is -O-, -NR ｇ - and -S-S- may be inserted; R ｇ is a hydrogen atom, C１－２０ Alkyl, Amino C ２－２０ Alkyl and hydroxy C ２－２０ alkyl, the alkyl portion of which may be inserted with a group selected from the group consisting of —O— and —NH—; Y ａ is C １－５ is alkylene; Y ｂ is C ２－８ Alkylene or C ２－８alkenylene; and m is an integer of 1 or more and 100 or less. (23) The method according to (21) or (22), wherein the introduction rate of the steryl group relative to the repeating units of disaccharides derived from the hyaluronic acid and the hyaluronic acid derivative is 25% or more and 60% or less. (24) The method according to (23), wherein the content of the cryoprotectant is 6 parts by mass or more and 1750 parts by mass or less relative to 10 parts by mass of the hyaluronic acid derivative component. (25) The method according to (21) or (22), wherein the introduction rate of the steryl group relative to the repeating units of disaccharides derived from the hyaluronic acid and the hyaluronic acid derivative is 0.1% or more and less than 25%. (26) The method according to (25), wherein the content of the cryoprotectant is 28 parts by mass or more and 1750 parts by mass or less relative to 10 parts by mass of the hyaluronic acid derivative component. (27) The molecular weight of the cryoprotectant is 200 or more. The method according to any one of (21) to (26). (28) The method according to any one of (21) to (26), wherein the cryoprotectant is a sugar or a sugar alcohol. (29) The method according to (28), wherein the sugar is one or more selected from the group consisting of sucrose, trehalose, and glucose. (30) The method according to (28) or (29), wherein the sugar alcohol is sorbitol. (31) The method according to (22), wherein R is a cholesteryl group or a phytosteryl group. (32) The method according to (31), wherein R is a cholesteryl group. (33) The method according to any one of (21) to (32), wherein the molecular weight of the hyaluronic acid derivative component is 1,000 or more and less than 20,000. (34) The method according to any one of (21) to (33), wherein, after the carrier composition is crushed in a mortar, 30% by mass or more of the powder passes through a sieve having an opening of 2.0 mm according to the dry sieving test method specified in JIS K 0069:1992.

[0011] The carrier composition of the above embodiment can prevent clogging of the filter during sterile filtration in the formulation.

[0012] Cholesteryl 6-aminohexylcarbamate hydrochloride obtained in step 1 of Production Example 1 １1H-NMR spectrum of the tetrabutylammonium (TBA) salt of hyaluronic acid (HA) obtained in step 2 of Production Example 1. １ 1H-NMR spectrum of the hyaluronic acid derivative (HA-C) into which 6-aminohexyl carbamate obtained in step 3 of Production Example 1 has been introduced. ６ -Chol) １ 1 is a H-NMR spectrum. It is a graph showing the relationship between the filtration resistance (Pa s / m) and the filtrate volume (mL) measured by a texture analyzer using each of the carrier compositions obtained in Examples 1 to 4, 8 to 10 and Comparative Examples 1 to 2, and 6 to 8. It is a graph showing the relationship between the filtration resistance (Pa s / m) and the filtrate volume (mL) measured by a texture analyzer using each of the carrier compositions obtained in Examples 5 to 7 and Comparative Examples 3 to 5. It is a graph showing the relationship between the filtration resistance (Pa s / m) and the filtrate volume (mL) measured by a texture analyzer using each of the carrier compositions obtained in Examples 11 and 12. ...-b. ６ -Chol) １ 1H-NMR spectrum. A graph showing the relationship between the filtration resistance (Pa s / m) and the filtrate volume (mL) measured by a texture analyzer using each carrier composition obtained in Examples 1-b to 3-b, 7-b to 10-b and Comparative Examples 1-b to 2-b, 6-b to 9-b. A graph showing the relationship between the filtration resistance (Pa s / m) and the filtrate volume (mL) measured by a texture analyzer using each carrier composition obtained in Examples 4-b to 6-b and Comparative Examples 3-b to 5-b. A graph showing the relationship between the filtration resistance (Pa s / m) and the filtrate volume (mL) measured by a texture analyzer using each carrier composition obtained in Examples 11-b to 12-b. The tetrabutylammonium (TBA) salt of hyaluronic acid (HA) obtained in step 2 in Production Example 1-c. １ 1H-NMR spectrum of the hyaluronic acid derivative (HA-C) into which 6-aminohexyl carbamate obtained in step 3 of Production Example 1-c has been introduced. ６ -Chol) １1H-NMR spectrum. It is a graph showing the relationship between the filtration resistance (Pa s / m) and the filtrate volume (mL) measured by a texture analyzer using each carrier composition obtained in Examples 1-c to 3-c, 9-c and Comparative Examples 1-c to 2-c, 8-c to 10-c. It is a graph showing the relationship between the filtration resistance (Pa s / m) and the filtrate volume (mL) measured by a texture analyzer using each carrier composition obtained in Examples 4-c to 6-c and Comparative Examples 3-c to 5-c. It is a graph showing the relationship between the filtration resistance (Pa s / m) and the filtrate volume (mL) measured by a texture analyzer using each carrier composition obtained in Examples 7-c to 8-c and Comparative Examples 6-c to 7-c.

[0013] The following describes in detail an embodiment of the present invention (hereinafter referred to as "the present embodiment"); however, the present invention is not limited to this embodiment, and various modifications are possible without departing from the spirit of the present invention.

[0014] The terms used in this specification will be explained below.

[0015] As used herein, "C １－２０ The term "alkyl" means a straight or branched chain alkyl group having from 1 to 20 carbon atoms, for example, methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, isobutyl, tert-butyl, etc. １－４ alkyl," and further includes n-pentyl, 3-methylbutyl, 2-methylbutyl, 1-methylbutyl, 1-ethylpropyl, n-hexyl, 4-methylpentyl, 3-methylpentyl, 2-methylpentyl, 1-methylpentyl, 3-ethylbutyl, 2-ethylbutyl, and the like. １－２０ Alkyl has 1 to 12 carbon atoms. １－１２ Alkyl, C having 1 to 6 carbon atoms １－６ Alkyl groups are also included.

[0016] As used herein, "C １－６ The term "alkylcarbonyl" refers to the alkyl moiety of the aforementioned C １－６It means an alkylcarbonyl group, for example, acetyl, propionyl, n-propylcarbonyl, isopropylcarbonyl, n-butylcarbonyl, sec-butylcarbonyl, isobutylcarbonyl, tert-butylcarbonyl, etc. １－４ alkylcarbonyl".

[0017] As used herein, "amino C ２－２０ The term "alkyl" means a linear or branched alkyl having from 2 to 20 carbon atoms and having an amino group as a substituent, and for example, the amino group may be located on the terminal carbon atom of the alkyl group. ２－２０ Alkyl includes amino C having 2 to 12 carbon atoms. ２－１２ Alkyl is also included.

[0018] As used herein, "hydroxy C ２－２０ The term "alkyl" means a linear or branched alkyl group having from 2 to 20 carbon atoms and having a hydroxy group as a substituent, for example, the hydroxy group may be located on the terminal carbon atom of the alkyl group. ２－２０ Alkyl includes hydroxy groups having 2 to 12 carbon atoms. ２－１２ Alkyl is also included.

[0019] As used herein, "C ２－３０ The term "alkylene" means a linear or branched divalent saturated hydrocarbon group having from 2 to 30 carbon atoms, and includes, for example, ethylene, propylene, etc., and has from 2 to 20 carbon atoms. ２－２０ Alkylene, C having 2 to 8 carbon atoms ２－８ Alkylene, group "-(CH ２ ) ｎ -" (where n is 2 or more and 30 or less, preferably 2 or more and 20 or less, and more preferably 2 or more and 15 or less).

[0020] As used herein, "C １－５ The term "alkylene" means a straight or branched chain saturated divalent hydrocarbon radical having from 1 to 5 carbon atoms and includes, for example, methylene, ethylene, propylene, and the like.

[0021] The term "C" referred to herein ２－８ The term "alkenylene" means a linear or branched, divalent saturated hydrocarbon group having from 2 to 8 carbon atoms and containing one or more double bonds, and examples thereof include -CH=CH-, -C(CH ３ )=CH-, 2-butene-1,4-diyl, hepta-2,4-diene-1,6-diyl, octa-2,4,6-triene-1,8-diyl, etc. When geometric isomers exist, each isomer and a mixture thereof are also included.

[0022] <<Carrier Composition>> Generally, a "carrier composition" is a composition used to encapsulate an active ingredient in the carrier composition, thereby solubilizing a poorly soluble active ingredient, inhibiting aggregation, imparting sustained release of the active ingredient, maintaining the activity of the active ingredient, delivering the active ingredient to target tissues or target cells in the body, etc. The carrier composition of this embodiment can be used in a formulation obtained by mixing the active ingredient in a form dissolved in a solvent and then sterilizing and filtering it. The carrier composition of this embodiment does not contain, for example, an active ingredient.

[0023] The carrier composition of this embodiment is a freeze-dried product, and contains a hyaluronic acid derivative component, which includes hyaluronic acid and a hyaluronic acid derivative with a steryl group introduced therein, and a cryoprotectant, and the hyaluronic acid derivative component and the cryoprotectant are mixed in an aqueous solution and then freeze-dried.That is, each powder particle of the freeze-dried product contains the hyaluronic acid derivative component and the cryoprotectant.

[0024] The hyaluronic acid derivative has a repeating unit represented by the following general formula (I):

[0025]

[0026] In the formula, R １ , R ２ , R ３ , and R ４ are each independently a hydrogen atom, C １－６ Alkyl, formyl and C １－６ alkylcarbonyl; Z represents a direct bond or a peptide linker consisting of any amino acid residues of 2 to 30; X １is represented by the following formula: ｂ -R, -NR ｂ -COO-R, -NR ｂ -CO-R, -NR ｂ -CO-NR ｃ -R, -COO-R, -O-COO-R, -SR, -CO-Y ａ -SR, -O-CO-Y ｂ -SR, -NR ｂ -CO-Y ｂ -SR, and -S-SR, a group selected from the group consisting of groups represented by R ａ , R ｂ and R ｃ are each independently a hydrogen atom, C １－２０ Alkyl, Amino C ２－２０ Alkyl and hydroxy C ２－２０ alkyl, where the alkyl portion of the group is selected from the group consisting of —O— and —NR ｆ - may be inserted with a group selected from the group consisting of; R ｆ is a hydrogen atom, C １－１２ Alkyl, Amino C ２－１２ Alkyl and hydroxy C ２－１２ alkyl, the alkyl portion of which may be interrupted by a group selected from the group consisting of -O- and -NH-; R is a steryl group; Y is C ２－３０ Alkylene, or -(CH ２ CH ２ O) ｍ -CH ２ CH ２ -, where the alkylene is -O-, -NR ｇ - and -S-S- may be inserted; R ｇ is a hydrogen atom, C １－２０ Alkyl, Amino C ２－２０ Alkyl and hydroxy C ２－２０ alkyl, the alkyl portion of which may be inserted with a group selected from the group consisting of —O— and —NH—; Y ａ is C １－５ is alkylene; Yｂ is C ２－８ Alkylene or C ２－８ alkenylene; and m is an integer of 1 or more and 100 or less.

[0027] In one aspect of the present invention, the introduction rate of the steryl group relative to the repeating units of the disaccharide derived from the hyaluronic acid and the hyaluronic acid derivative is 25% or more and 60% or less, preferably 25% or more and 55% or less, more preferably 25% or more and 50% or less, and even more preferably 25% or more and 45% or less (hereinafter, the carrier composition of this aspect is referred to as the "first carrier composition of this embodiment"). With the steryl group introduction rate within the above range, the hyaluronic acid derivative component forms stable microparticles even under physiological salt concentrations, and has the property of being stably dispersed in water. Furthermore, with the steryl group introduction rate within the above range, the hyaluronic acid derivative component can be complexed with an active ingredient such as a drug and administered to the body, for example, intravenously, to produce a preparation containing a high concentration of a poorly soluble active ingredient, a solution preparation in which the active ingredient is stably dispersed, a blood-sustained release preparation, a preparation in which the activity of the active ingredient is maintained for a long period of time, and a preparation targeting the target tissue or cell, etc.

[0028] In one aspect of the present invention, the introduction rate of the steryl group relative to the repeating unit of the disaccharide derived from the hyaluronic acid and the hyaluronic acid derivative is 0.1% or more but less than 25%, preferably 5% or more but less than 25%, more preferably 6% or more but less than 22%, and even more preferably 6% or more but less than 20% (hereinafter, the carrier composition of this aspect is referred to as the "second carrier composition of this embodiment"). When the introduction rate of the steryl group is within the above range, the hyaluronic acid derivative component has the property of being well dissolved in pure water or under low salt concentration, and flocculating and forming precipitate under physiological salt concentration. In addition, when the introduction rate of the steryl group is within the above range, the hyaluronic acid derivative component can be complexed with an active ingredient and administered to the body, for example, by subcutaneous administration, to obtain a precipitation-type sustained-release preparation that takes advantage of the characteristic of flocculating after administration.

[0029] The steryl group introduction rate is １ It can be measured by H-NMR measurement. １The integral value of the peak derived from the steryl group of the hyaluronic acid derivative in the H-NMR spectrum and the peak derived from the acetyl group of N-acetyl-D-glucosamine contained in the hyaluronic acid derivative (COCH ３ , 1.6 ppm or more and 2.0 ppm or less, 3H) and the integral value of Ｈ represents the number of hydrogen atoms corresponding to the peak. Specifically, the measurement can be carried out, for example, according to the method described in the Examples below.

[0030] "Steryl group introduction rate" (%) = [(steryl group-derived peak integral value × 3 / n Ｈ ) / (peak integral value derived from the acetyl group of N-acetyl-D-glucosamine) × 100

[0031] In the first carrier composition of this embodiment, the content of the cryoprotectant is 6 parts by mass or more and 1750 parts by mass or less, preferably 6 parts by mass or more and 1000 parts by mass or less, more preferably 6 parts by mass or more and 100 parts by mass or less, even more preferably 6 parts by mass or more and 80 parts by mass or less, particularly preferably 8 parts by mass or more and 80 parts by mass or less.In the second carrier composition of this embodiment, the content of the cryoprotectant is 28 parts by mass or more and 1750 parts by mass or less, preferably 28 parts by mass or more and 1000 parts by mass or less, more preferably 28 parts by mass or more and 200 parts by mass or less, even more preferably 28 parts by mass or more and 185 parts by mass or less, particularly preferably 30 parts by mass or more and 185 parts by mass or less, relative to 10 parts by mass of the hyaluronic acid derivative component. When the content of the cryoprotectant is equal to or greater than the lower limit, as shown in the examples described below, when the lyophilized carrier composition is dissolved in a solvent and sterile filtered, the filtration resistance does not increase significantly, and sterile filterability can be improved. That is, filter clogging during sterile filtration during formulation can be suppressed. On the other hand, when the content is equal to or less than the upper limit, the aqueous solution after dissolving the lyophilized carrier composition becomes a more hypotonic solution, and pharmaceutical additives can be further added during formulation.

[0032] The carrier composition of the present embodiment may contain salt, but from the viewpoint of suppressing the aggregation and precipitation of hyaluronic acid derivative components in the preparation before being administered to living body, its content is the amount that becomes a concentration that is sufficiently lower than physiological salt concentration when dissolved in water.In addition, "physiological salt concentration" here means the salt concentration at the administration site of the animal that is the administration target.For example, the physiological salt concentration in human is the concentration of sodium chloride of about 150mM.In addition, "low salt concentration" means a concentration that is sufficiently lower than physiological salt concentration.

[0033] The salt may be derived from the raw materials of the hyaluronic acid derivative component and the cryoprotectant, or may be added separately to adjust the osmotic pressure of the solution when dissolved in a solvent such as water. Examples of the salt that can be contained in the carrier composition of this embodiment include, but are not limited to, sodium salt, potassium salt, magnesium salt, calcium salt, etc.

[0034] Therefore, by having the above-mentioned configuration, the carrier composition of this embodiment can suppress filter clogging during sterile filtration during formulation, which was a problem with conventional carrier compositions containing hyaluronic acid derivative components.

[0035] Next, each of the components contained in the carrier composition of this embodiment will be described in detail below.

[0036] <Hyaluronic Acid Derivative Component> The hyaluronic acid derivative component includes hyaluronic acid and a hyaluronic acid derivative.

[0037] In the hyaluronic acid derivative, the steryl group may be directly bound to the hyaluronic acid or may be bound via a linker.

[0038] The "linker" referred to here can be any peptide linker or synthetic compound linker that can be introduced by genetic engineering.The length of peptide linker is not particularly limited, and those skilled in the art can appropriately select it according to purpose, but the preferred length is 2 amino acids or more, while the upper limit is not particularly limited, but it is usually 30 amino acids or less, preferably 20 amino acids or less, and particularly preferably 15 amino acids.The carbon number of synthetic compound linker is not particularly limited, and those skilled in the art can appropriately select it according to purpose, but the preferred carbon number is 2 or more, while the upper limit is not particularly limited, but it is usually 20 or less, preferably 12 or less.The linkers contained in hyaluronic acid derivative may all be the same length or carbon number linkers, or may be different lengths or carbon number linkers.

[0039] [Steryl Group] The term "steryl group" used herein is not particularly limited as long as it is a group having a steroid skeleton. Specific examples of steroids include cholesterol, cholestanol, campestanol, ergostanol, stigmastanol, coprostanol, stigmasterol, sitosterol, lanosterol, ergosterol, cimialenol, bile acids, testosterone, estradiol, progesterone, cortisol, cortisone, aldosterone, corticosterone, and deoxycortisterone. Examples of steryl groups include cholesteryl, stigmasteryl, lanosteryl, and ergosteryl groups. Of these, cholesteryl groups (particularly, cholest-5-en-3β-yl groups) and phytosteryl groups are preferred, with cholesteryl groups being more preferred.

[0040] The hyaluronic acid derivative has one or more repeating units represented by the following general formula (I) (hereinafter, sometimes referred to as "repeating unit (I)").

[0041]

[0042] In the formula, R １ , R ２ , R ３ , and R ４ are each independently a hydrogen atom, C１－６ Alkyl, formyl and C １－６ alkylcarbonyl; Z represents a direct bond or a peptide linker consisting of any amino acid residues of 2 to 30; X １ is represented by the following formula: ｂ -R, -NR ｂ -COO-R, -NR ｂ -CO-R, -NR ｂ -CO-NR ｃ -R, -COO-R, -O-COO-R, -SR, -CO-Y ａ -SR, -O-CO-Y ｂ -SR, -NR ｂ -CO-Y ｂ -SR, and -S-SR, a group selected from the group consisting of groups represented by R ａ , R ｂ and R ｃ are each independently a hydrogen atom, C １－２０ Alkyl, Amino C ２－２０ Alkyl and hydroxy C ２－２０ alkyl, where the alkyl portion of the group is selected from the group consisting of —O— and —NR ｆ - may be inserted with a group selected from the group consisting of; R ｆ is a hydrogen atom, C １－１２ Alkyl, Amino C ２－１２ Alkyl and hydroxy C ２－１２ alkyl, the alkyl portion of which may be interrupted by a group selected from the group consisting of -O- and -NH-; R is a steryl group; Y is C ２－３０ Alkylene, or -(CH ２ CH ２ O) ｍ -CH ２ CH ２ -, where the alkylene is -O-, -NR ｇ - and -S-S- may be inserted; R ｇ is a hydrogen atom, C １－２０ Alkyl, Amino C ２－２０ Alkyl and hydroxy C２－２０ alkyl, the alkyl portion of which may be inserted with a group selected from the group consisting of —O— and —NH—; Y ａ is C １－５ is alkylene; Y ｂ is C ２－８ Alkylene or C ２－８ alkenylene; and m is an integer of 1 or more and 100 or less.

[0043] In particular, the hyaluronic acid derivative preferably includes a hyaluronic acid derivative having one or more repeating units represented by the following general formula (Ia) (hereinafter, sometimes referred to as "repeating unit (Ia)").

[0044]

[0045] In the formula, R １ , R ２ , R ３ , and R ４ are each independently a hydrogen atom, C １－６ Alkyl, formyl and C １－６ alkylcarbonyl; X is selected from the group consisting of -NR ａ -Y-NR ｂ is a hydrophobic group represented by —COO—R; R ａ and R ｂ are each independently a hydrogen atom and C １－６ alkyl; R is a steryl group; Y is C ２－３０ Alkylene, or -(CH ２ CH ２ O) ｍ -CH ２ CH ２ -, and m is an integer between 1 and 100.

[0046] Here, when the hyaluronic acid derivative contains two or more repeating units (I) or two or more repeating units (Ia), the repeating units may be the same or different.

[0047] The hyaluronic acid derivative may be modified at a position other than the repeating unit (I) or the repeating unit (Ia), for example, the hydroxy group may be modified by —O(C １－６ alkyl), -O(formyl), -O(C １－６ The carboxy group may be converted into an amide or ester, or may form a salt.

[0048] [Repeating unit (I)] The group "-Z-N(R ａ ) Y-X １ " is a group represented by the following formula: -NH-(CH ２ ) ｍｚ -NH-R; -NH-(CH ２ ) ｍｚ -NH-COO-R; -NH-(CH ２ CH ２ O) ｍ -CH ２ CH ２ -NH-COO-R; -NH-(CH ２ ) ｍｚ -COO-R; -NH-(CH ２ CH ２ O) ｍ -CH ２ CH ２ -COO-R, -NH-(CH ２ ) ｍｚ -O-COO-R; -NH-(CH ２ CH ２ O) ｍ -CH ２ CH ２ -O-COO-R, -NH-(CH ２ ) ｍｚ -SR; -NH-(CH ２ CH ２ O) ｍ -CH ２ CH ２ -SR; -NH-(CH ２ ) ｍｚ -O-CO-CH(R ８ )-CH ２ -SR; -NH-(CH ２ ) ｍｚ -NHCO-CH(R ８ )-CH２ -SR; -NH-(CH ２ CH ２ O) ｍ -CH ２ CH ２ -NHCO-CH(R ８ )-CH ２ -SR; -NH-(CH ２ CH ２ O) ｍ -CH ２ CH ２ -O-CO-CH(R ８ )-CH ２ -SR; -NH-(CH ２ ) ｍｚ -S-S-R; and -Z-NR ａ -Y-NR ｂ -COO-R; where mz is an integer of 2 or more and 30 or less, and R ８ is a hydrogen atom or a methyl group, and R and m are as defined previously in this specification; and ２ ) ｍｚ -NH-COO-R; -NH-(CH ２ CH ２ O) ｍ -CH ２ CH ２ -NH-COO-R; and -NH-(CH ２ ) ｍｚ Preferred are groups selected from the group consisting of: -S-S-R; where mz, R and m are as previously defined herein.

[0049] (Z) In general formula (I), Z is preferably a direct bond. In another embodiment, when Z is a peptide linker, X １ Ha-NR ｂ In another embodiment, Z is preferably —NH—[CH(-Z ａ )-CONH] ｎ－１ -CH(-Z ａ )-CO-, where n is an integer of 2 to 30, and Z ａ are each independently H２ N-CH(-Z ａ The peptide linker is attached to the carboxy group of the glucuronic acid moiety at the N-terminus and to the group —N(—R )—COOH at the C-terminus. ａ )-Y-X １ Examples of amino acids that can be used as amino acid residues of the peptide linker include α-amino acids, such as natural (L-form) amino acids such as alanine, arginine, asparagine (Asn), aspartic acid, cysteine, glutamine, glutamic acid, glycine (Gly), histidine, isoleucine, leucine (Leu), lysine, methionine, phenylalanine (Phe), proline, serine, threonine, tryptophan, tyrosine, and valine, as well as D-forms thereof, and all α-amino acids, including synthetic amino acids, can be used. ａ Examples of the group include -CH ３ , H ２ NC(NH)NH(CH ２ ) ３ -, H ２ NCOCH ２ -, etc. Furthermore, n Z's may be the same or different. n is an integer of 2 or more and 30 or less, preferably 2 or more and 10 or less, and more preferably 2 or more and 4 or less. Preferred examples of peptide linkers include -Gly-Phe-Leu-Gly-, -Asn-Phe-Phe-, -Phe-Phe-, Phe-Gly-, etc.

[0050] (Y) In the general formula (I), Y is —(CH ２ ) ｎ１ - and - (CH ２ CH ２ O) ｍ１ -CH ２ CH ２ - (where n1 is an integer of 2 or more and 20 or less, preferably an integer of 2 or more and 15 or less, more preferably an integer of 2 or more and 12 or less, and even more preferably an integer of 2 or more and 6 or less; m1 is an integer of 1 or more and 4 or less) is preferred. Specifically, -(CH ２ ) ２ -, -(CH ２) ６ -, -(CH ２ ) ８ -, -(CH ２ ) １２ - or -(CH ２ CH ２ O) ２ -CH ２ CH ２ From the viewpoint of realizing high solubility in pure water or under low salt concentrations, Y is preferably -(CH ２ ) ２ -, -(CH ２ ) ６ -, -(CH ２ ) ８ - and - (CH ２ ) １２ - is preferably a group selected from the group consisting of -(CH ２ ) ６ - is more preferable.

[0051] Y is, for example, —CH ２ CH ２ O-CH ２ CH ２ -S-S-CH ２ CH ２ O-CH ２ CH ２ -, -(CH ２ CH ２ O) ２ -CH ２ CH ２ -S-S-CH ２ CH ２ O-CH ２ CH ２ -, -CH ２ CH ２ O-CH ２ CH ２ -S-S-(CH ２ CH ２ O) ２ -CH ２ CH ２ -, -(CH ２ CH ２ O) ２ -CH ２ CH ２ -S-S-(CH ２ CH ２ O) ２ -CH２ CH ２ - etc.

[0052] (Y ａ ) Y ａ As the group, -CH ２ - or -CH ２ -CH ２ - is preferred.

[0053] (Y ｂ ) Y ｂ As the group, -CH ２ -CH ２ -, -CH(CH ３ ) CH ２ -, 2-butene-1,4-diyl, hepta-2,4-diene-1,6-diyl or octa-2,4,6-triene-1,8-diyl is preferred, and -CH ２ -CH ２ - or -CH(CH ３ ) CH ２ - is more preferable.

[0054] Group “-Z-N(R ａ ) Y-X １ " is exemplified by -NH-(CH ２ ) ２ -NH-CO-cholesteryl, -NH-(CH ２ ) ４ -NH-(CH ２ ) ３ -NH-(CH ２ ) ３ -NH-COO-cholesteryl, -NH-(CH ２ ) ３ -NH-(CH ２ ) ４ -NH-(CH ２ ) ３ -NH-COO-cholesteryl, -NH-(CH ２ ) ４ -NH-(CH ２ ) ３ -NH-COO-cholesteryl, -NH-(CH ２ ) ４ -N(-(CH ２ ) ３ -NH ２ )-COO-cholesteryl, -NH-(CH ２ )３ -NH-(CH ２ ) ４ -N(-(CH ２ ) ３ -NH ２ )-COO-cholesteryl, -NH-(CH ２ ) ３ -NH-(CH ２ ) ４ -N(-(CH ２ ) ３ -NH-(CH ２ ) ３ -NH ２ )-COO-cholesteryl, -NH-(CH ２ ) ３ -NH-(CH ２ ) ４ -N(-(CH ２ ) ３ -NH ２ )-CO-NH-cholesteryl, -NH-(CH ２ ) ３ -NH-(CH ２ ) ４ -N(-(CH ２ ) ３ -NH ２ )-CO-cholesteryl, -NH-(CH ２ ) ３ -NH-(CH ２ ) ４ -N(-(CH ２ ) ３ -NH ２ )-cholesteryl, etc. Preferred groups include "-Z-N(R ａ ) Y-X １ " as R ａ , R ｂ and R ｃ is a hydrogen atom, and Y is a linear C ２－３０ Alkylene or -(CH ２ CH ２ O) ｍ -CH ２ CH ２ - and Y ａ But linear C １－５ alkylene, or Y ｂ But linear C ２－８ Alkylene or linear C ２－８ It is alkenylene.

[0055] [Repeating unit (Ia)] In general formula (Ia), X is —NH—(CH ２ ) ２ -NH-COO-cholesteryl, -NH-(CH ２ ) ６ -NH-COO-cholesteryl, -NH-(CH ２ ) １２ -NH-COO-cholesteryl or -NH-(CH ２ CH ２ O) ２ -CH ２ CH ２ -NH-COO-cholesteryl is preferred, and -NH-(CH ２ ) ２ -NH-COO-cholesteryl, -NH-(CH ２ ) ６ -NH-COO-cholesteryl or -NH-(CH ２ CH ２ O) ２ -CH ２ CH ２ --NH--COO-cholesteryl is more preferred.

[0056] In addition to the repeating unit (I), the hyaluronic acid derivative may further contain a repeating unit represented by general formula (II) (hereinafter, sometimes referred to as "repeating unit (II)").

[0057]

[0058] In the formula, R １ａ , R ２ａ , R ３ａ , and R ４ａ are each independently a hydrogen atom, C １－６ Alkyl, formyl and C １－６ alkylcarbonyl; X ａ is hydroxy and -O-Q ＋ wherein Q is selected from the group consisting of ＋ is the countercation.

[0059] Here, when hyaluronic acid derivative contains two or more repeating units (II), these repeating units may be the same or different.In another embodiment, hyaluronic acid derivative may be the hyaluronic acid derivative that is essentially composed of repeating units (I), repeating units (Ia) and repeating units (II).

[0060] [Repeating unit (II)] In general formula (II), Q ＋ is not particularly limited as long as it is a counter cation that forms a salt with a carboxy group in water, and when it is divalent or higher, it forms a salt with multiple carboxy groups depending on the valence. Examples of the counter cation include metal ions such as lithium ion, sodium ion, rubidium ion, cesium ion, magnesium ion, and calcium ion; ＋ R ｊ R ｋ R ｌ R ｍ (In the formula, R ｊ , R ｋ , R ｌ and R ｍ are each independently a hydrogen atom and C １－６ Among them, ammonium ions represented by Q ＋ is preferably a sodium ion, a potassium ion, or a tetraalkylammonium ion (for example, a tetra-n-butylammonium ion). ｊ , R ｋ , R ｌ and R ｍ is C １－６ Preferably, they are identical groups selected from the group consisting of alkyl, and preferably n-butyl groups.

[0061] R １ , R ２ , R ３ , and R ４ , and R １ａ , R ２ａ , R ３ａ , and R ４ａ are preferably all hydrogen atoms. ａ and R ｂ are preferably all hydrogen atoms.

[0062] Among these, the hyaluronic acid derivative is preferably a hyaluronic acid derivative substantially consisting of repeating units (I) and (II). In the hyaluronic acid derivative, for example, 80% or more, preferably 90% or more, and more preferably 95% or more of the repeating units of the disaccharide consisting of D-glucuronic acid and N-acetyl-D-glucosamine contained in the derivative are repeating units (I) and (II). The hyaluronic acid derivative may be composed only of repeating units (I) and (II).

[0063] In the first carrier composition of this embodiment, the content of the hyaluronic acid derivative component is not particularly limited, but can be, for example, 0.57 parts by mass or more and 62.5 parts by mass or less, 0.99 parts by mass or more and 62.5 parts by mass or less, 9.1 parts by mass or more and 62.5 parts by mass or less, 10.8 parts by mass or more and 62.5 parts by mass or less, 11.1 parts by mass or more and 62.5 parts by mass or less, or 11.1 parts by mass or more and 55.5 parts by mass or less, per 100 parts by mass of the carrier composition.

[0064] In the second carrier composition of this embodiment, the content of the hyaluronic acid derivative component is not particularly limited, but can be, for example, 0.57 parts by mass or more and 26.3 parts by mass or less, 0.99 parts by mass or more and 26.3 parts by mass or less, 4.8 parts by mass or more and 26.3 parts by mass or less, 4.8 parts by mass or more and 25.0 parts by mass or less, or 5.1 parts by mass or more and 25.0 parts by mass or less, per 100 parts by mass of the carrier composition.

[0065] [the manufacturing method of hyaluronic acid derivatives] As the manufacturing method of hyaluronic acid derivatives, for example, the carboxyl group of glucuronic acid is converted into amide, and steryl group is introduced, thereby obtaining hyaluronic acid derivatives.In addition, by adjusting the compound amount of the compound that has steryl group that reacts with raw material hyaluronic acid or its derivative, the introduction rate of steryl group can be adjusted.

[0066] Specific examples of the method for converting the carboxy group of glucuronic acid into an amide and introducing a hydrophobic group, for example, a steryl group, include ion-exchanging a starting material, hyaluronic acid or a derivative thereof, preferably hyaluronic acid or a derivative thereof composed only of the repeating unit (II), with a tetraalkylammonium salt (for example, a tetrabutylammonium (TBA) salt), and then condensing the hyaluronic acid salt with a compound of the formula: "HNR ａ -Y-NR ｂ -R, NHR ａ -Y-NR ｂ -COO-R, HNR ａ -Y-NR ｂ -COO-R, HNR ａ -Y-NR ｂ -CO-R, HNR ａ -Y-NR ｂ -CO-NR ｃ -R, HNR ａ -Y-COO-R, HNR ａ -Y-O-COO-R, HNR ａ -Y-S-R, HNR ａ -Y-CO-Y ａ -S-R, HNR ａ -Y-O-CO-Y ｂ -S-R, HNR ａ -Y-NR ｂ -CO-Y ｂ -S-R, HNR ａ -Y-S-S-R or -Z-NR ａ -Y-NR ｂ -COO-R (in the formula, R ａ , R ｂ , R ｃ , Y, Y ａ , Y ｂ , Z and R are as defined above) with an amine having introduced therein a steryl group, particularly cholesteryl, represented by the formula (I)

[0067] The condensing agent that can be used in the above reaction is not particularly limited, and examples thereof include 4-(4,6-dimethoxy-1,3,5-triazine)-4-methylmorpholinium (DMT-MM), N,N'-carbonyldiimidazole (CDI), N,N'-dicyclohexylcarbodiimide (DCC), N-ethoxycarbonyl-2-ethoxy-1,2-dihydroquinoline (EEDQ), 2-benzotriazole-1,1,3,3-tetramethyluronium tetrafluoroborate (TBTU), 3,4-dihydro-3-hydroxy-4-oxo-1,2,3-benzotriazine (HODhbt), benzotriazole-1-oxy-tris-pyrrolidino-phosphonium hexafluorophosphate (PyBOP), and benzotriazole-1-yl-oxy-tris(dimethylamino)phosphonium. Examples include hexafluorophosphate (BOP), 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDC), and N-hydroxysuccinimide (NHS).

[0068] In particular, although not limited to, DMT-MM is preferred in that the reaction proceeds highly efficiently even in a mixed solvent of water and an organic solvent. Furthermore, by using DMT-MM as a condensing agent, in a system in which a large number of hydroxy groups coexist, it is possible to suppress the formation of ester bonds and to highly selectively form amide bonds between amino groups and carboxy groups. The use of this condensing agent can, for example, prevent the reaction of the solvent alcohol with the carboxy group of the hyaluronic acid moiety, or prevent the formation of undesired crosslinks due to intramolecular or intermolecular bonding between the carboxy group and hydroxy groups simultaneously present in the hyaluronic acid moiety.

[0069] Examples of solvents used in the steryl group introduction reaction include water, DMSO, methanol, ethanol, propanol, butanol, isopropanol, polyhydric alcohols, acetonitrile, DMF, THF, dichloromethane, chloroform, hexane, diethyl ether, ethyl acetate, and mixed solvents thereof. The polyhydric alcohol may be a dihydric alcohol or a trihydric alcohol. Examples of dihydric alcohols include ethylene glycol, diethylene glycol, propylene glycol, dipropylene glycol, neopentyl glycol, 1,4-butanediol, and 1,6-hexanediol. Examples of trihydric alcohols include glycerin and trimethylolpropane.

[0070] Alternatively, the starting hyaluronic acid or a derivative thereof may be ion-exchanged with a tetraalkylammonium salt (e.g., tetrabutylammonium (TBA) salt), and the hyaluronic acid salt may be reacted with a spacer moiety in a solvent in the presence of a suitable condensing agent (protection and deprotection reactions may be carried out as necessary), converting the carboxy group (-COOH) of the starting hyaluronic acid or a derivative thereof, followed by reaction with a suitable reagent. Examples of combinations of groups derived from carboxy groups and reaction reagents are shown below. -CONR ａ -Y-NR ｂ H + Hal-R; -CONR ａ -Y-NR ｂ H + Hal-COOR; -CONR ａ -Y-NR ｂ H + HOCO-R; -CONR ａ -Y-NR ｂ H + Hal-CO-R; -CONR ａ -Y-NR ｂ -COOH + HNR ｃ -R; -CONR ａ -Y-NR ｂ -CO-NR ｃ H + Hal-R; -CONR ａ -Y-NR ｂ H + HOCO-NR ｃ -R; -CONR ａ -Y-NRｂ H + Hal-CO-NR ｃ -R; -CONR ａ -Y-COOH + HO-R; -CONR ａ -Y-OH + Hal-COO-R; -CONR ａ -Y-OCOOH + HO-R; -CONR ａ -Y-OCOOH + Hal-R; -CONR ａ -Y-OCO-Hal + HO-R; -CONR ａ -Y-SH + Hal-R; -CONR ａ -Y-Hal + HS-R; -CONR ａ -Y-CO-Y ａ -Hal + HS-R; -CONR ａ -Y-CO-Y ａ -SH + Hal-R; -CONR ａ -Y-O-CO-CH=CH ２ + HS-R; -CONR ａ -Y-NR ｂ -CO-CH(CH ３ ) = CH ２ + HS-R; -CONR ａ -Y-SH + HS-R; -COZ-OH + HNR ａ -Y-NR ｂ -COO-R; -COZ-NR ａ -Y-NR ｂ H + Hal-COO-R; where R ａ , R ｂ , R ｃ , Y, Y ａ , Y ｂ and Z are as previously defined herein, and Hal represents a halogen atom selected from the group consisting of a fluorine atom, a chlorine atom, a bromine atom and an iodine atom.

[0071] Examples of reaction modes include dehydrohalogenation reactions, condensation reactions, dehydration reactions, nucleophilic addition reactions such as Michael addition, and oxidative disulfide formation reactions. These are well-known reactions, and can be appropriately selected by those skilled in the art and carried out under preferred reaction conditions. When the conversion product or reactant has a carboxy group, it may be converted into an N-hydroxysuccinimide (hereinafter also referred to as "NHS") ester and then reacted.

[0072] Another method is to react 2-aminoethyl 2-pyridyl disulfide with the carboxyl group of the starting hyaluronic acid or its derivative to prepare a hyaluronic acid derivative into which a spacer having a mercapto group modified with a leaving group is introduced at the end, and then to subject this to a nucleophilic substitution reaction with thiocholesterol to form a disulfide bond.

[0073] Another example is a method in which a hyaluronic acid or derivative thereof in which a part of a spacer is introduced into the carboxy group and a part of a spacer is introduced into the steryl group are prepared and then reacted. Some specific examples have been described above, but when -S-S- is inserted into Y, another example is a method in which a hyaluronic acid derivative in which a spacer having a terminal mercapto group is introduced into the carboxy group of hyaluronic acid and a steryl group in which a spacer having a terminal mercapto group is introduced are prepared, and then these are oxidatively reacted to form a disulfide bond. In this case, one mercapto group can be reacted with 2-mercaptopyridine to form a disulfide, and then substituted with the other mercapto group.

[0074] After preparing the hyaluronic acid derivative, other substituents may be further introduced. For example, in a hyaluronic acid derivative essentially consisting of the repeating unit (I) and the repeating unit (II), 0.1% to 99.5%, preferably 40% to 65%, of the carboxy groups may be replaced with -CO-X ｚ , [where X ｚ is the following group: -NH-(CH ２ ) ｐ１ —O—CO—C(R １７ ) = CH ２ -NH-(CH ２ ) ｐ１-O-CO-CH(R １７ )-CH ２ -S-CH ２ -CH(OH)-CH(OH)-CH ２ -SH; -NH-(CH ２ ) ｐ１ -SH; -NH-(CH ２ ) ｐ１ -NH-CO-C(R １７ )=CH ２ ; -NH-(CH ２ ) ｐ１ -NH-C(=NH)-(CH ２ ) ３ -SH; -NH-(CH ２ ) ｐ１ -NH-CO-(CH ２ ) ｒ -SH; -NH-(CH ２ ) ｐ１ -NH-CO-CH(R １７ )-CH ２ -S-CH ２ -CH(OH)-CH(OH)-CH ２ -SH; -NH-(CH ２ ) ｐ１ -NH-CO-CH(NH ２ )-CH ２ -SH; -NH-(CH ２ ) ｐ１ -NH-CO-CH(NH ２ )-(CH ２ ) ２ -SH; -NH-NH-CO-(CH ２ ) ４ -CO-NH-NH-C(=NH)-(CH ２ ) ３ -SH; -NH-(CH ２ -CH ２ -O) ｑ -CH ２ -CH ２ -O-CO-C(R １７ )=CH ２ ; -NH-(CH ２ -CH ２ -O) ｑ -CH ２ -CH ２ -O-CO-CH(R １７)-CH ２ -S-CH ２ -CH(OH)-CH(OH)-CH ２ -SH; -NH-(CH ２ -CH ２ -O) ｑ -CH ２ -CH ２ -SH; -NH-(CH ２ -CH ２ -O) ｑ -CH ２ -CH ２ -NH-CO-C(R １７ )=CH ２ ; -NH-(CH ２ -CH ２ -O) ｑ -CH ２ -CH ２ -NH-C(=NH)-(CH ２ ) ３ -SH; -NH-(CH ２ -CH ２ -O) ｑ -CH ２ -CH ２ -NH-CO-(CH ２ ) ｒ -SH; -NH-(CH ２ -CH ２ -O) ｑ -CH ２ -CH ２ -NH-CO-CH(R １７ )-CH ２ -S-CH ２ -CH(OH)-CH(OH)-CH ２ -SH; -NH-(CH ２ -CH ２ -O) ｑ -CH ２ -CH ２ -NH-CO-CH(NH ２ )-CH ２ -SH; -NH-(CH ２ -CH ２ -O) ｑ -CH ２ -CH ２ -NH-CO-CH(NH ２ )-(CH ２ ) ２-SH; -NH-CH(CO ２ H)-(CH ２ )-SH; -NH-CH(CO ２ H)-(CH ２ ) ２ -SH; and -NH-CH(CO ２ H)-(CH ２ ) ２ -CONH-CH(CONH-CH ２ -CO ２ H) —CH ２ -SH; where R １７ is a hydrogen atom or C １－６ wherein p1 is an alkyl group selected from the group consisting of: p1 is an integer of 2 or more and 10 or less, q is an integer of 1 or more and 200 or less, and r is an integer of 1 or more and 3 or less;

[0075] <Cryoprotective Agent> The cryoprotective agent is not particularly limited as long as it is known as a "cryoprotective agent" or a "lyoprotective agent," and examples thereof include sugars, sugar alcohols, dextran, polyethylene glycol, propylene glycol, glycerin, glycerol, polyvinylpyrrolidone, dimethyl sulfoxide, etc. Among these, sugars or sugar alcohols are preferred, and sugars are more preferred.

[0076] The molecular weight of cryoprotectant is not particularly limited, and can be suitably selected.The molecular weight of cryoprotectant is, for example, more than 100, preferably more than 200, more preferably more than 250, and even more preferably more than 300.When the molecular weight of cryoprotectant is above the lower limit, the aqueous solution after dissolving the carrier composition as freeze-dried product becomes more hypotonic, and can further incorporate pharmaceutical additives during formulation.In addition, the molecular weight of cryoprotectant is, for example, below 1,000, preferably below 800, more preferably below 600, and even more preferably below 400.When the molecular weight of cryoprotectant is below the upper limit, if necessary, during formulation, it is easier to separate and remove only the cryoprotectant from the aqueous solution after dissolving the carrier composition as freeze-dried product, without removing hyaluronic acid derivative components, by using membrane separation techniques such as ultrafiltration membrane.

[0077] Preferred examples of sugars include monosaccharides and disaccharides, and specific examples include monosaccharides such as glucose, fructose, mannose, and galactose; and disaccharides such as sucrose, lactulose, lactose, maltose, trehalose, cellobiose, kojibiose, nigerose, isomaltose, isotrehalose, neotrehalose, sophorose, laminaribiose, gentiobiose, turanose, maltulose, palatinose, gentiobiulose, mannobiose, melibiose, melibiulose, neolactose, galactosucrose, scillabiose, neohesperidose, rutinose, rutinulose, vicianose, xylobiose, and primeverose. Among these, glucose, sucrose, or trehalose is preferred in the first carrier composition of this embodiment, and disaccharides are preferred, with sucrose or maltose being more preferred in the second carrier composition of this embodiment.

[0078] Examples of sugar alcohols include sorbitol, erythritol, xylitol, lactitol, mannitol, etc. Among these, sorbitol is preferred.

[0079] Among these, glucose, sucrose, or trehalose are preferred in the first carrier composition of this embodiment because they are widely used as cryoprotectants, and sucrose, maltose, or sorbitol are preferred in the second carrier composition of this embodiment, with sucrose being more preferred because of its proven use as a pharmaceutical additive.

[0080] <Method for producing carrier composition> The carrier composition of this embodiment can be obtained by a production method including, for example, the following steps (A) to (C): step (A) of adjusting the concentration of the hyaluronic acid derivative component in the aqueous solution to 4 mg / mL or more and 100 mg / mL or less; step (B) of adding a cryoprotectant to the aqueous solution after step (A); and step (C) of freeze-drying the aqueous solution containing the hyaluronic acid derivative component and the cryoprotectant.

[0081] Each step will be described in detail below.

[0082] [Step (A)] In step (A), adjust the concentration of hyaluronic acid derivative components in aqueous solution to be more than 4mg / mL and less than 100mg / mL.By the concentration of hyaluronic acid derivative components being more than above-mentioned lower limit, as shown in the example below, when the carrier composition of freeze-dried body is dissolved in solvent and sterilized filtered, filtration resistance does not increase significantly, and can make sterilization filterability good.On the other hand, by the concentration of hyaluronic acid derivative components being less than above-mentioned upper limit, can prevent the poor drying caused by the thawing of the aqueous solution that comprises hyaluronic acid derivative components and cryoprotectant in the step (C) below.

[0083] From the viewpoint of ensuring the fluidity of aqueous solution, the concentration of hyaluronic acid derivatives components is preferably below 100mg / mL, more preferably below 50mg / mL, and even more preferably below 25mg / mL.On the other hand, from the viewpoint of suppressing the clogging of filter during sterilization filtration during preparation, in the first carrier composition of this embodiment, the concentration of hyaluronic acid derivatives components is preferably above 4mg / mL, and more preferably above 7mg / mL.In addition, in the second carrier composition of this embodiment, the concentration of hyaluronic acid derivatives components is preferably above 4.0mg / mL, and more preferably above 5.0mg / mL.

[0084] The aqueous solution of hyaluronic acid derivative components can be prepared by, for example, dissolving or dispersing the hyaluronic acid derivative components in solid or powder form in a solvent that contains water, so that the concentration of hyaluronic acid derivative components is within the above-mentioned numerical range.Alternatively, when the concentration of hyaluronic acid derivative components in the aqueous solution of hyaluronic acid derivative components is lower than the above-mentioned numerical range, it can be concentrated, and when the concentration of hyaluronic acid derivative components in the aqueous solution of hyaluronic acid derivative components is higher than the above-mentioned numerical range, it can be diluted.

[0085] The method for preparing the aqueous solution of the hyaluronic acid derivative component so that the concentration of the hyaluronic acid derivative component falls within the above-mentioned numerical range is not particularly limited, and examples thereof include a method of adjusting the concentration of the raw material hyaluronic acid or its derivative when reacting hyaluronic acid or its derivative with a compound having a steryl group, and a method of adjusting the amount of the hyaluronic acid derivative component added when producing the aqueous solution of the hyaluronic acid derivative component.

[0086] The method of concentration operation is not particularly limited, and for example, microfiltration, ultrafiltration, reverse osmosis, vacuum evaporation, freeze concentration, reverse osmosis concentration, etc. can be used. Among them, from the viewpoint of preventing the decomposition of the hyaluronic acid derivative, it is preferable to use microfiltration, ultrafiltration, or reverse osmosis.

[0087] The method of dilution is not particularly limited, as long as it is a method of using aqueous solution to dilute composition and adjust the concentration of hyaluronic acid derivative components.As the aqueous solution for dilution, for example, water, phosphate buffer solution, histidine buffer solution, citrate buffer solution, glucose aqueous solution, sucrose aqueous solution, maltose aqueous solution etc. can be used.Among them, from the viewpoint of preventing hyaluronic acid derivative from flocculating and precipitating, water or the aqueous solution that does not contain inorganic salt is preferred.In addition, the aqueous solution for dilution can be used with the aqueous solution of cryoprotectant, and the step (B) described below can be carried out simultaneously.

[0088] The concentration of the hyaluronic acid derivative in the aqueous solution of the hyaluronic acid derivative component can be measured, for example, by size exclusion chromatography (SEC). Specifically, it can be measured by the method described in the Examples below.

[0089] [Step (B)] In step (B), add cryoprotectant to the aqueous solution after step (A).By adding cryoprotectant, the existence of cryoprotectant can inhibit the formation of ice crystals when the aqueous solution is frozen, and the conformational change of hyaluronic acid derivatives components caused by contact with the hydrophobic surface of ice crystals can be suppressed, so that the dispersion stability of the particles formed by hyaluronic acid derivatives components is not impaired.

[0090] In step (B), the cryoprotectant may be added in a solid state or in a state dissolved in a solvent such as water.

[0091] In the first carrier composition of this embodiment, the amount of cryoprotectant added is 6 parts by mass or more and 1750 parts by mass or less, preferably 6 parts by mass or more and 1000 parts by mass or less, more preferably 6 parts by mass or more and 100 parts by mass or less, even more preferably 6 parts by mass or more and 80 parts by mass or less, particularly preferably 8 parts by mass or more and 80 parts by mass or less.In the second carrier composition of this embodiment, the amount of cryoprotectant added is 28 parts by mass or more and 1750 parts by mass or less, preferably 28 parts by mass or more and 1000 parts by mass or less, more preferably 28 parts by mass or more and 200 parts by mass or less, even more preferably 28 parts by mass or more and 185 parts by mass or less, even more preferably 30 parts by mass or more and 185 parts by mass or less, particularly preferably 30 parts by mass or more and 80 parts by mass or less. When the amount of cryoprotectant added is equal to or greater than the lower limit, as shown in the examples described below, when the lyophilized carrier composition is dissolved in a solvent and sterile filtered, the filtration resistance does not increase significantly, and sterile filterability can be improved. That is, filter clogging during sterile filtration during formulation can be suppressed. On the other hand, when the amount is equal to or less than the upper limit, the aqueous solution after dissolving the lyophilized carrier composition becomes a more hypotonic solution, and pharmaceutical additives can be further added during formulation.

[0092] As the cryoprotectant, the same ones as those exemplified above in the "cryoprotectant" can be used.

[0093] [Step (C)] In step (C), an aqueous solution containing a hyaluronic acid derivative component and a cryoprotectant (hereinafter, sometimes referred to as "aqueous solution containing a hyaluronic acid derivative component and a cryoprotectant") is freeze-dried.

[0094] The freeze-drying method is not particularly limited, as long as it freezes an aqueous solution containing a hyaluronic acid derivative component and a cryoprotectant, and then reduces the pressure while maintaining the frozen state, thereby sublimating and removing the water content in the aqueous solution. For example, step (C) can include the following steps (C1) to (C4). In this case, if it is desired to increase the dryness of the freeze-dried product, it is preferable to include step (C4). Conversely, if it is desired to retain the bound water in the freeze-dried product, it is preferable not to include step (C4). Step (C1) (also referred to as the "addition step"): adding an aqueous solution containing a hyaluronic acid derivative component and a cryoprotectant to a container in the device; Step (C2) (also referred to as the "pre-freezing step"): cooling and pre-freezing the aqueous solution; Step (C3) (also referred to as the "primary drying step"): reducing the pressure while maintaining the aqueous solution in a frozen state to remove ice crystals contained in the aqueous solution; and Step (C4) (also referred to as the "secondary drying step"): further increasing the temperature after the primary drying is completed to increase the dryness of the composition.

[0095] The apparatus used for freeze-drying is not particularly limited, and for example, a commercially available freeze-dryer can be used. Among them, from the viewpoint of controlling the degree of vacuum, a freeze-dryer capable of monitoring the degree of vacuum within the apparatus during freeze-drying is preferred, and from the viewpoint of controlling the product temperature, a freeze-dryer capable of monitoring the product temperature within the apparatus during freeze-drying is preferred.

[0096] In the method for producing a carrier composition, step (C) preferably includes one or more steps selected from the group consisting of steps (Ca) and (Cb) shown below: step (Ca) of adjusting the thickness of the aqueous solution containing a hyaluronic acid derivative component and a cryoprotectant at the start of lyophilization to 10 mm or more and 50 mm or less; step (Cb) of adjusting the degree of vacuum to 5 Pa or more and less than 100 Pa during lyophilization.

[0097] [Step (Ca)] In step (Ca), the thickness of the aqueous solution containing a hyaluronic acid derivative component and a cryoprotectant is adjusted to 10 mm or more and 50 mm or less. Here, the "thickness of the aqueous solution" refers to the vertical thickness of the aqueous solution containing a hyaluronic acid derivative component and a cryoprotectant placed in a container before freezing.

[0098] As thickness adjustment method, for example, in the above-mentioned step (C1) (introducing step), the method of adjusting the amount of sample solution to be introduced into container can be mentioned.The thickness of aqueous solution is preferably 10mm or more and 50mm or less, more preferably 10mm or more and 40mm or less, even more preferably 10mm or more and 30mm or less, particularly preferably 10mm or more and 20mm or less, and most preferably 13mm or more and 17mm or less.By making the thickness of aqueous solution above-mentioned lower limit value or more, it can suppress the increase in particle size of the microparticles formed by hyaluronic acid derivative component during freeze-drying.On the other hand, by making the thickness of aqueous solution below above-mentioned upper limit value, it can more effectively prevent the drying failure caused by the melting of hyaluronic acid derivative component and the aqueous solution containing cryoprotectant during freeze-drying.

[0099] [Step (Cb)] In step (Cb), the degree of vacuum is adjusted to 5 Pa or more and less than 100 Pa during freeze-drying.

[0100] Specifically, in step (C-b), the pressure in the container is reduced from atmospheric pressure to a vacuum, and then the pressure is reduced to a vacuum of 5 Pa or more and less than 100 Pa (hereinafter sometimes referred to as a "specific vacuum range") while maintaining the aqueous solution in a frozen state. Among these, step (C-b) is preferably performed in step (C3) (primary drying step). It is also preferable to maintain the vacuum within a specific vacuum range at least until the product temperature matches the shelf temperature.

[0101] The degree of vacuum is preferably 5 Pa or more and less than 100 Pa, more preferably 5 Pa or more and 40 Pa or less, even more preferably 10 Pa or more and 40 Pa or less, and particularly preferably 20 Pa or more and 40 Pa or less.By making the degree of vacuum equal to or greater than the lower limit, it can suppress the increase in the particle size of the microparticles that hyaluronic acid derivative components form during freeze-drying.On the other hand, by making the degree of vacuum less than the upper limit, it can more effectively prevent the poor drying caused by the melting of the aqueous solution that contains hyaluronic acid derivative components and cryoprotectant during freeze-drying.

[0102] Examples of methods for controlling the degree of vacuum include a method of adjusting the amount of aqueous solution to be added in the above step (C1) (adding step), a method of adding water or another aqueous solution to a container separate from the aqueous solution to adjust the total amount of liquid in the device, etc. Alternatively, a vacuum control function of the freeze dryer to be used may be used.

[0103] <Characteristics of the carrier composition> [Average particle size] In the first carrier composition of this embodiment, the average particle size of the spherical structures containing a complex of a cryoprotectant and a hyaluronic acid derivative can be 25 nm or more and 75 nm or less. In the second carrier composition of this embodiment, the average particle size of the spherical structures containing a complex of a cryoprotectant and a hyaluronic acid derivative can be 50 nm or more and 150 nm or less. The average particle size of the carrier composition of this embodiment can be measured, for example, by DLS (Dynamic Light Scattering), a nanotracking particle measuring device, size exclusion chromatography, high performance liquid chromatography, electron microscopy, etc. More specifically, for example, the hyaluronic acid derivative is diluted with 10 mM phosphate buffer containing 10 w / v% sucrose to a concentration of 1 mg / mL using a DLS device and measured.

[0104] [Molecular weight] The molecular weight of hyaluronic acid derivative component is not particularly limited, but it is preferable to select the molecular weight according to the function that hyaluronic acid derivative is expected to have.For example, in the first carrier composition of this embodiment, when final dosage form is solution preparation (liquid preparation) and expects good syringeability, the hyaluronic acid derivative with relatively small molecular weight is preferred, and the molecular weight of this hyaluronic acid derivative is preferably 1,000 or more and less than 20,000.In addition, in the second carrier composition of this embodiment, when expecting the sustained release function derived from the diffusion delay in local administration, or when expecting the ability to form precipitates under physiological salt concentration, the hyaluronic acid derivative with relatively large molecular weight is preferred.The molecular weight of this hyaluronic acid derivative is preferably 1,000 or more and less than 1,000,000, more preferably 5,000 or more and less than 300,000, more preferably 10,000 or more and less than 120,000, and particularly preferably 20,000 or more and less than 50,000. The molecular weight of the hyaluronic acid derivative can generally be adjusted by using a raw material having a corresponding molecular weight.

[0105] The "molecular weight of the hyaluronic acid derivative" referred to here is the weight-average molecular weight determined by size exclusion chromatography multi-angle light scattering detector (SEC-MALS). Specifically, it can be measured according to the method described in the Examples below.

[0106] [Filtration Resistance] The first carrier composition of the present embodiment has a filtration resistance of 1.8 × 10 when the filtrate volume is increased from 0 mL to 15 mL, as measured under the following conditions: ９ It is preferable that the viscosity is less than Pa·s / m, and 1.5×10 ９ It is more preferable that the viscosity is 8.0×10 Pa·s / m or less. ８ It is more preferable that the viscosity is 4.0×10 Pa·s / m or less. ８ The second carrier composition of the present embodiment has a filtration resistance of 1.8×10 to 15 mL of filtrate volume, measured under the following conditions: ９ It is preferable that the viscosity is less than 1.6×10 Pa·s / m. ９It is more preferable that the viscosity is 1.4×10 Pa·s / m or less. ９ It is more preferable that the viscosity is not more than Pa·s / m. When the filtration resistance from 0 mL to 15 mL of filtrate is less than the upper limit or not more than the upper limit, the sterile filterability can be improved. In other words, filter clogging during sterile filtration in formulation can be suppressed.

[0107] Specifically, the filtration resistance until the filtrate volume reaches 15 mL is measured by first dissolving the hyaluronic acid derivative component contained in the first carrier composition of this embodiment in water to a concentration of 12 mg / mL, or by dissolving the hyaluronic acid derivative component contained in the second carrier composition of this embodiment in water to a concentration of 5.0 mg / mL, followed by stirring overnight. Then, a syringe filter (Merck; SLGP033NS; Millex-GP, 0.22 μm, PES, 33 mm; membrane area 4.5 cm) is placed in a Terumo syringe (Terumo; SS-20ESZ; capacity 20 mL) containing 15 mL to 20 mL of the aqueous solution of the carrier composition. ２ ) is attached to the test piece and placed in a texture analyzer, and the indentation test force (N) is measured under the following measurement conditions.

[0108] (Measurement conditions) Texture analyzer: Shimadzu Corporation EZ Test series EZ-SX Jig: Shimadzu Corporation Shimadzu Autograph Syringe push-out test force test jig 346-57828 Syringe push-in speed: 10 mm / min

[0109] Next, the filtration resistance is calculated using the measured indentation test force (N) according to the following formula.

[0110] (Calculation formula for filtration resistance) "Filtration pressure (Pa)" = Test force (N) / Filtration area (m ２ ) "Filtration speed (m / s)" = filtration amount (m ３ ) / filtration area (m ２ ) time (s) derivative "filtration resistance (Pa m / s)" = filtration pressure (Pa) / filtration rate (m / s) = test force (N) / (syringe cross-sectional area (m ２ ) × syringe pushing speed (m / s)

[0111] [2 mm sieve under-sieve mass fraction after pulverization process] The carrier composition of this embodiment may be in the form of a cake or a powder. After pulverization in a mortar, the carrier composition of this embodiment has a 2.0 mm mesh size passing sieve according to the dry sieving test method specified in JIS K 0069:1992 (hereinafter also referred to as "2 mm sieve under-sieve mass fraction after pulverization process") of preferably 30 mass% or more, more preferably 40 mass% or more, even more preferably 70 mass% or more, and particularly preferably 95 mass% or more. When the 2 mm sieve under-sieve mass fraction after pulverization is the above lower limit or more, the carrier composition is pulverized in a mortar to form a powder, which makes it easier to handle.

[0112] Specifically, the method for measuring the mass fraction (%) under a 2 mm sieve after the pulverization step involves first weighing 100 to 110 mg of the lyophilized product and placing it in a magnetic mortar (medium-capacity magnetic mortar set, 130 g, porcelain; 1-6022-02; AS ONE). Next, the product is placed in a stirrer (magnetic mortar stirrer MMPS-T1; 1-6016-11; AS ONE) and pulverized at 60 rpm for 1 minute. The pulverized sample is then recovered and its mass is measured to confirm that the recovered mass is 70% or more of the mass weighed in the mortar. Next, dry sieving is performed using a 2.0 mm sieve (manufactured by SANPO) based on the dry sieving test method specified in JIS K 0069:1992. The masses of the sample on and under the sieve are measured using an electronic balance. The undersieve mass fraction is then calculated using the following formula: (Formula for calculating undersieve mass fraction) "Undersieve mass fraction (%)" = undersieve sample mass (mg) / (on-sieve sample mass (mg) + undersieve sample mass (mg)) x 100

[0113] Pharmaceutical Composition The carrier composition of the above embodiment can be prepared as a pharmaceutical composition by mixing the carrier composition with an active ingredient in a dissolved form in a solvent and then sterilizing and filtering the mixture. That is, in one embodiment, the present invention provides a pharmaceutical composition obtained by sterilizing and filtering a mixture of the active ingredient and the carrier composition.

[0114] The hyaluronic acid derivative component in the carrier composition forms a nano-sized hydrogel by the self-association of the steryl groups in the hyaluronic acid derivative in water, resulting in the association of single or multiple molecules. Thus, in a formulation incorporating the carrier composition of this embodiment and an active ingredient, the active ingredient and the hyaluronic acid derivative form a complex (hereinafter, sometimes referred to as an "active ingredient-hyaluronic acid derivative complex"). Specifically, the steryl groups in the hyaluronic acid derivative and the active ingredient form a complex through hydrophobic interaction, presumably exhibiting a core-shell-like spherical structure in which the active ingredient and hydrophobic moieties, such as the steryl groups, are present in the center, while hydrophilic moieties, such as hyaluronic acid and moieties derived from hyaluronic acid in the hyaluronic acid derivative, are present in the outer periphery. In other words, it is presumed that the active ingredient is encapsulated or encapsulated in the hyaluronic acid derivative.

[0115] It is preferred that the hyaluronic acid derivative component and the active ingredient in the carrier composition are directly or indirectly bound to each other to form a conjugate, and are not in a free state. That is, it is preferred that one or more active ingredients are bound to the hyaluronic acid derivative component in the pharmaceutical composition to form a hyaluronic acid derivative-active ingredient conjugate.

[0116] When the pharmaceutical composition of this embodiment is administered into the body, the active ingredient is gradually released from the carrier composition, and can be expected to have good sustained release.The bond between the hyaluronic acid derivative component and the active ingredient can be either covalent or non-covalent, but from the viewpoint of maintaining the activity of the active ingredient, non-covalent bond is preferred.

[0117] In the pharmaceutical composition comprising the first carrier composition of this embodiment, hyaluronic acid derivative component forms stable microparticles even under physiological salt concentration, and has the property of being stable and dispersible in water, so that the pharmaceutical composition comprising the first carrier composition of this embodiment is useful as blood sustained-release preparation and target tissue or cell targeting preparation.In addition, because the hyaluronic acid derivative component has the above-mentioned property, the pharmaceutical composition comprising the first carrier composition of this embodiment can make poorly soluble active ingredient stably disperse in vivo.Therefore, the pharmaceutical composition comprising the first carrier composition of this embodiment is useful as the sustained-release agent for poorly soluble active ingredient.

[0118] In the pharmaceutical composition comprising the second carrier composition of this embodiment, hyaluronic acid derivative component has the ability to form precipitate under physiological salt concentration.Therefore, when the pharmaceutical composition comprising the second carrier composition of this embodiment is a dispersible microparticle solution, it is in a solution state before being administered into the body, but can be a precipitate-type sustained-release preparation with the feature of being in-situ aggregated after being administered into the body by administering (for example, subcutaneously).In addition, because the hyaluronic acid derivative component has the above-mentioned properties, when the pharmaceutical composition comprising the second carrier composition of this embodiment is a precipitate suspension, it can be a precipitate-type sustained-release preparation characterized by the active ingredient being difficult to burst release.In this case, since it can also be given syringeability, the size of precipitate is preferably 200 μ m or less, more preferably 100 μ m or less, and even more preferably 20 μ m or less.

[0119] When the pharmaceutical composition of this embodiment is dissolved or dispersed in solvent, it is considered that the hydrophobic interaction between the steryl group of hyaluronic acid derivative component and the active ingredient present in the system will spontaneously combine to form the conjugate of the active ingredient and hyaluronic acid derivative component.By forming this conjugate, it is expected that the storage stability of the active ingredient will improve, the biological activity will be maintained, the sustained release will improve, the solubility in water will improve, the resistance to the stimuli such as heat and light will improve, and the aggregation and precipitation will be suppressed.

[0120] <Active ingredient> The active ingredient is not particularly limited as long as it is a component that can be used as a pharmaceutical for humans or animals, and examples thereof include proteins, peptides, nucleic acids, and low molecular weight compounds.

[0121] Examples of low molecular weight compounds include anticancer agents such as alkylating agents, antimetabolites, and alkaloids; immunosuppressants; anti-inflammatory agents such as steroids and non-steroidal anti-inflammatory agents; antirheumatic agents; and antibacterial agents such as β-lactam antibiotics, aminoglycoside antibiotics, macrolide antibiotics, tetracycline antibiotics, new quinolone antibiotics, and sulfa drugs.

[0122] Examples of proteins and peptides include erythropoietin (EPO), granulocyte colony-stimulating factor (G-CSF), interferon-α, β, γ (INF-α, β, γ), thrombopoietin (TPO), serial neutrophic factor (CNTF), tumor necrosis factor (TNF), tumor necrosis factor binding protein (TNFbp), interleukin-10 (IL-10), FMS-like tyrosine kinase (Flt-3), growth hormone (GH), insulin, insulin-like growth factor-1 (IGF-1), platelet-derived growth factor (PDGF), interleukin-1 receptor agonist (IL-10), and IL-1 receptor agonist (IL-10). Examples of such antibodies include scepter antagonist (IL-1ra), brain-derived neurotrophic factor (BDNF), keratinocyte growth factor (KGF), stem cell factor (SCF), megakaryocyte growth differentiation factor (MGDF), osteoprotegerin (OPG), leptin, parathyroid hormone (PTH), basic fibroblast growth factor (b-FGF), bone morphogenetic protein (BMP), atrial natriuretic peptide (ANP), brain natriuretic peptide (BNP), C-type natriuretic peptide (CNP), glucagon-like peptide-1 (GLP-1), antigens for vaccines, antibodies, diabodies, minibodies, and fragmented antibodies.

[0123] Examples of nucleic acids include DNA, RNA, antisense nucleic acids, decoy nucleic acids, ribozymes, small interfering RNAs, and nucleic acid aptamers.

[0124] The pharmaceutical composition of this embodiment is administered, for example, orally, parenterally, intranasally, intravaginally, intraocularly, subcutaneously, intravenously, intramuscularly, intradermally, intraperitoneally, intracerebrally, or intraorally.

[0125] The pharmaceutical composition of this embodiment is preferably a liquid preparation dissolved or dispersed in a pharmaceutically acceptable solvent or the like, and more preferably a dispersible microparticle solution or a precipitated suspension, and can be used in the form of, for example, an injection, a patch preparation, a microneedle preparation, a topical ointment, eye drops, a spray, an inhalant, or the like.

[0126] When the pharmaceutical composition comprising the first carrier composition of this embodiment is a liquid, the concentration of hyaluronic acid derivative component in the pharmaceutical composition is preferably 0.01mg / mL or more and 500mg / mL or less, more preferably 0.1mg / mL or more and 200mg / mL or less, more preferably 0.2mg / mL or more and 100mg / mL or less, particularly preferably 0.5mg / mL or more and 50mg / mL or less.The concentration of hyaluronic acid derivative component in the pharmaceutical composition is above the lower limit, so that the storage stability of the active ingredient is improved, the maintenance of biological activity, the improvement of sustained release, and when the active ingredient is poorly water-soluble, the solubilization of the active ingredient in water etc. can be improved, and the dosage of the active ingredient tends to be increased.On the other hand, the concentration of hyaluronic acid derivative component in the pharmaceutical composition is below the upper limit, so that the injectability when administered into the living body using a syringe needle is improved, and the sterilization filterability tends to be improved.

[0127] When the pharmaceutical composition containing the second carrier composition of this embodiment is a liquid, the concentration of the hyaluronic acid derivative component in the pharmaceutical composition is preferably 1 mg / mL or more and 200 mg / mL or less, more preferably 4 mg / mL or more and 100 mg / mL or less, even more preferably 4 mg / mL or more and 50 mg / mL or less, and particularly preferably 4 mg / mL or more and 12 mg / mL or less.

[0128] The concentration of hyaluronic acid derivative components in pharmaceutical composition is above above-mentioned lower limit, can make the precipitation ability under physiological salt concentration more excellent, and can also increase the dosage of drug.On the other hand, the concentration of hyaluronic acid derivative components in pharmaceutical composition is below above-mentioned upper limit, can improve the injectability when using injection needle to administer into living body, and can improve the sterilization filterability.

[0129] <Method for producing pharmaceutical composition> The pharmaceutical composition of this embodiment can be produced by dissolving the carrier composition in a solvent such as water or a known buffer solution, mixing the mixture with an active ingredient, and sterilizing by filtration.

[0130] When the carrier composition and the active ingredient are mixed, pharmaceutically acceptable additives may be mixed together. Examples of pharmaceutically acceptable additives include carriers other than the above-mentioned carrier composition, solubilizers, stabilizers, isotonicity agents, buffers, pH adjusters, preservatives, bactericides or antibacterial agents, thickeners, etc.

[0131] Examples of carriers other than the carrier composition include aqueous solvents such as water and aqueous ethanol.

[0132] Examples of solubilizers include polyoxyethylene hydrogenated castor oil, polyoxyl 40 stearate, povidone, polysorbate 80, and the like.

[0133] Examples of stabilizers include sodium edetate hydrate, polyvinylpyrrolidone (povidone), polysorbate 80, and the like.

[0134] Examples of isotonic agents include potassium chloride, calcium chloride, sodium chloride, concentrated glycerin, glucose, D-mannitol, and the like.

[0135] Examples of buffering agents include sodium citrate hydrate, sodium bicarbonate, dry sodium carbonate, sodium carbonate, magnesium sulfate, trometamol, boric acid, borax, sodium hydrogen phosphate hydrate, sodium dihydrogen phosphate, and potassium dihydrogen phosphate.

[0136] Examples of pH adjusters include hydrochloric acid and sodium hydroxide.

[0137] Examples of the antiseptic, disinfectant or antibacterial agent include benzalkonium chloride, methyl parahydroxybenzoate, propyl parahydroxybenzoate, chlorobutanol, sorbic acid, alkylpolyaminoethylglycine and the like.

[0138] Examples of thickening agents include carboxyvinyl polymer, povidone, polyvinyl alcohol (partially saponified), hydroxyethyl cellulose, hydroxypropyl cellulose, hydroxypropylmethyl cellulose, hypromellose, methyl cellulose, and glycerin.

[0139] Sterile filtration is carried out once or repeatedly using a membrane with a pore size of, for example, 0.10 μm to 0.45 μm. Sterile filtration can also be carried out by connecting two or more filters having pore sizes in the above range in series. Commonly used membrane materials include cellulose derivatives such as cellulose acetate; polyvinyl chloride, PVDF, PES, nylon, etc., but other materials can also be used.

[0140] In the method for producing a pharmaceutical composition of this embodiment, the carrier composition is dissolved in a solvent and mixed with an active ingredient, and then the sterile filtration is performed, thereby preventing a significant increase in filtration resistance and improving sterile filtration performance. That is, filter clogging during sterile filtration during formulation can be suppressed. Therefore, the method for producing a pharmaceutical composition of this embodiment can also be said to be a method for suppressing filter clogging during sterile filtration during formulation, which includes mixing the carrier composition dissolved in a solvent with an active ingredient, and then sterile filtration to form a formulation.

[0141] The present invention will be described in detail below with reference to examples, but these examples are not intended to limit the scope of the present invention.

[0142] The methods for measuring and evaluating the physical properties of the "aqueous solution of hyaluronic acid derivative component" produced in the production example and the "carrier composition" produced in the examples and comparative examples are as follows.

[0143] <Methods for Measuring Physical Properties> [Physical Property 1] (Molecular Weight of Hyaluronic Acid Derivative Component) The molecular weight of the hyaluronic acid derivative component is the weight-average molecular weight determined by size exclusion chromatography multi-angle light scattering detector (SEC-MALS). A lyophilized carrier composition (20 mg of hyaluronic acid derivative component content) was dissolved in ultrapure water (10 mL) and stirred at room temperature for 12 hours or more to obtain an aqueous solution containing the hyaluronic acid derivative component (2 mg / mL of hyaluronic acid derivative component). To this aqueous solution containing the hyaluronic acid derivative component (750 μL), a 650 mM aqueous solution of hydroxypropyl-β-cyclodextrin (HP-β-CD) (750 μL) was added, mixed for 10 seconds using a shaker, and incubated at 37°C for 1 hour. The resulting sample was then subjected to SEC-MALS measurement to determine the weight-average molecular weight. The SEC-MALS measurement conditions are shown below.

[0144] (Measurement conditions) Column: TSKgel GMPWXL (manufactured by Tosoh Corporation) x 2 Column temperature: 30°C Eluent: phosphate buffered saline (pH 7.4) containing 10 mM HP-β-CD Flow rate: 1 mL / min Injection volume: 200 μL

[0145] [Physical Property 2] (Steryl Group Introduction Rate) The steryl group introduction rate of the hyaluronic acid derivative component is １ This was determined by H-NMR measurement. ６ A measurement solvent was prepared by mixing 20% ​​deuterium carbonate (99.9 v / v%, containing 0.05 v / v% trimethylsilyl (TMS), manufactured by Fujifilm Wako Co., Ltd.) and 20% deuterium carbonate (99.5 v / v%, manufactured by Fujifilm Wako Co., Ltd.) in a mass ratio of 99:1. Subsequently, a freeze-dried carrier composition (containing 2 mg of hyaluronic acid derivative components) was added to this measurement solvent (0.6 mL), and the mixture was treated in an ultrasonic bath for 30 minutes to completely dissolve the carrier composition. １ The product was subjected to H-NMR measurement. １ H-NMR measurements were carried out using a Fourier transform nuclear magnetic resonance (FT-NMR) spectrometer (ECS400, manufactured by JEOL Ltd.) at a sample temperature of 85°C. The steryl group introduction rate was evaluated by measuring the peak (COCH ３, 1.6 ppm to 2.0 ppm, 3H) and the peak derived from the methyl group in the cholesteryl group (CH ３ The introduction rate of cholesteryl groups into hyaluronic acid units was calculated from the integrals of the peaks (5H, 0.7 ppm, 3H) derived from cholesteryl groups using the following formula: Note that, because the peak derived from the acetyl group of N-acetyl-D-glucosamine overlaps with the peak derived from the cholesteryl group (5H) in the vicinity of 1.6 ppm to 2.0 ppm, which includes the peak derived from the acetyl group of N-acetyl-D-glucosamine, the value calculated by subtracting 5 / 3 times the integral of the peak derived from the cholesteryl group methyl (0.7 ppm) from the integral of the peak in the vicinity of 1.6 ppm to 2.0 ppm (i.e., integral (1.6 ppm to 2.0 ppm) - integral (0.7 ppm) × 5 / 3) was used as the integral of the peak derived from the acetyl group of N-acetyl-D-glucosamine to calculate the steryl group introduction rate.

[0146] [Steryl group introduction rate (%)] = [(Integrated value of peak derived from methyl group in cholesteryl group) / (Integrated value of peak derived from acetyl group of N-acetyl-D-glucosamine)] × 100 = [Integrated value (0.7 ppm) / {Integrated value (1.6 ppm or more and 2.0 ppm or less) - Integral value (0.7 ppm) × 5 / 3}] × 100

[0147] [Property 3] (Concentration of Hyaluronic Acid Derivative Component) The concentration of the hyaluronic acid derivative component contained in the aqueous solution of the hyaluronic acid derivative component was determined by size exclusion chromatography (SEC). The aqueous solution of the hyaluronic acid derivative component was diluted with an appropriate amount of ultrapure water to adjust the concentration of the hyaluronic acid derivative component to be in the range of 1.25 mg / mL to 2.5 mg / mL. To this aqueous solution of the hyaluronic acid derivative component (750 μL), a 650 mM hydroxypropyl-β-cyclodextrin (HP-β-CD) aqueous solution (750 μL) was added, mixed for 10 seconds using a shaker, and incubated at 37 ° C for 1 hour to prepare an unknown sample. Subsequently, lyophilized carrier composition (25 mg each) was dissolved in ultrapure water (20 mL) and ultrapure water (10 mL), respectively, and stirred at room temperature for 12 hours or more to prepare standard solutions of hyaluronic acid derivative components (concentrations of hyaluronic acid derivative components: 1.25 mg / mL and 2.5 mg / mL). 650 mM hydroxypropyl-β-cyclodextrin (HP-β-CD) aqueous solution (750 μL each) was added to the standard solutions of hyaluronic acid derivative components (750 μL each), mixed for 10 seconds using a shaker, and incubated at 37 ° C for 1 hour to prepare standard samples. The obtained unknown samples and standard samples were then filtered through a 0.45 μm syringe filter and subjected to SEC measurement, and the area value of the peak derived from the hyaluronic acid derivative components was calculated. From the area value of standard sample, make calibration curve, and calculate the concentration of hyaluronic acid derivatives that contain in unknown sample based on this calibration curve, thereby determine the concentration of hyaluronic acid derivatives that contain in the aqueous solution of hyaluronic acid derivatives.The conditions of SEC measurement are as follows:

[0148] (SEC measurement conditions) Column: TSKgel G3000SWXL (manufactured by Tosoh Corporation) Column temperature: 30°C Eluent: phosphate buffered saline (pH 7.4) containing 10 mM HP-β-CD Flow rate: 1 mL / min Injection volume: 50 μL Detector: RI

[0149] <Evaluation Methods> [Evaluation 1] (Sterile Filterability) The sterile filterability of the carrier composition was measured by dissolving the carrier composition in water (concentration of hyaluronic acid derivative component: 12 mg / mL), stirring overnight, and filtering the solution through a sterile filter. The filtration resistance was measured as follows: A Terumo syringe (manufactured by Terumo; SS-20ESZ; capacity 20 mL) containing 15 mL to 20 mL of an aqueous solution of the carrier composition was inserted into a syringe filter (manufactured by Merck; SLGP033NS; Millex-GP, 0.22 μm, PES, 33 mm; membrane area 4.5 cm). ２ The indentation test force (N) was measured under the following measurement conditions. The conditions for the texture analyzer measurement are shown below.

[0150] (Measurement conditions) Texture analyzer: Shimadzu Corporation EZ Test series EZ-SX Jig: Shimadzu Corporation Shimadzu Autograph Syringe push-out test force test jig 346-57828 Syringe push-in speed: 10 mm / min

[0151] Next, the filtration resistance was calculated using the measured indentation test force (N) according to the following formula.

[0152] (Calculation formula for filtration resistance) "Filtration pressure (Pa)" = Test force (N) / Filtration area (m ２ ) "Filtration speed (m / s)" = filtration amount (m ３ ) / filtration area (m ２ ) time (s) derivative "filtration resistance (Pa m / s)" = filtration pressure (Pa) / filtration rate (m / s) = test force (N) / (syringe cross-sectional area (m ２ ) × syringe pushing speed (m / s)

[0153] Based on the filtration resistance calculated as above, the presence or absence of a significant increase in the filtration resistance was evaluated by judging based on the following criteria.

[0154] (Evaluation criteria: Examples 1 to 12, Comparative Examples 1 to 8, Examples 1-c to 9-c, Comparative Examples 1-c to 10-c) A: The filtration resistance did not increase significantly (the filtration resistance increased to 1.8 × 10 before the amount of filtrate reached 15 mL). ９(Pa·s / m)) A-: Although it corresponds to A, a slight increase in filtration resistance was observed (it corresponds to A and the filtration resistance did not reach 8.0×10 ８ B: A significant increase in filtration resistance was observed (the filtration resistance reached 1.8 × 10 before the filtrate volume reached 15 mL). ９ (Pa s / m)

[0155] (Evaluation criteria: Examples 1-b to 12-b, Comparative Examples 1-b to 9-b) A': The filtration resistance did not increase significantly (the filtration resistance was 8.0 × 10 ８ (Pa·s / m)) A'-: Although it corresponds to A', a slight increase in filtration resistance was observed (it corresponds to A' and the filtration resistance did not reach 4.0 × 10 ８ B': A significant increase in filtration resistance was observed (the filtration resistance reached 8.0 × 10 before the amount of filtrate reached 15 mL). ８ (Pa s / m)

[0156] [Evaluation 2] (Osmotic pressure) Osmotic pressure (mOsm / L) was calculated based on the following formula. In the following formula, the concentration (mg / L) of cryoprotectant is the concentration contained in the solution obtained by dissolving carrier composition in water (concentration of hyaluronic acid derivative component: 12 mg / mL).

[0157] "Osmolarity (mOsm / L)" = concentration of cryoprotectant (mg / L) / molecular weight of cryoprotectant

[0158] Based on the osmotic pressure calculated as above, the solution was evaluated as to whether it was hypotonic or not by judging it based on the following criteria.

[0159] (Evaluation criteria) A: Hypotonic solution (osmotic pressure less than 285 mOsm / L) B: Not hypotonic solution (osmotic pressure 285 mOsm / L or more)

[0160] [Evaluation 3] (Unsieved Mass Fraction After Pulverization) The unsieved mass fraction (%) after the pulverization process was evaluated by the following procedure. First, 100-110 mg of the freeze-dried product was weighed and placed in a magnetic mortar (medium-capacity magnetic mortar set 130 g porcelain; 1-6022-02; AS ONE), and then placed in a stirrer (magnetic mortar stirrer MMPS-T1; 1-6016-11; AS ONE) and pulverized at 60 rpm for 1 minute. The pulverized sample was recovered and its mass was measured. It was confirmed that the recovered mass was 70% or more of the mass weighed in the mortar. Next, dry sieving was performed using a 2.0 mm mesh sieve (SANPO) based on the JIS K 0069:1992 test method. The masses of the sample on and under the sieve were measured using an electronic balance. Then, the undersize mass fraction was calculated using the following formula.

[0161] (Formula for calculating undersieve mass fraction) "Undersieve mass fraction (%)" = undersieve sample mass (mg) / (on-sieve sample mass (mg) + undersieve sample mass (mg)) x 100

[0162] (Evaluation criteria: crushability) A: The mass fraction (%) of the particles that remain after sieving after mortar crushing is 30% or more. B: The mass fraction (%) of the particles that remain after mortar crushing is less than 30%.

[0163] <Production of aqueous solution of hyaluronic acid derivative component> [Production Example 1] (hyaluronic acid derivative component HA-C ６ Preparation of aqueous solution of hyaluronic acid derivative component HA-C ６ An aqueous solution of -Chol was prepared according to the following steps 1 to 3.

[0164] 1. Step 1: Synthesis of cholesteryl 6-aminohexylcarbamate hydrochloride Cholesteryl 6-aminohexylcarbamate hydrochloride (Chol-C ６ Hydrochloride) was synthesized according to the following steps 1-1 and then 1-2.

[0165] (1) Step 1-1: To a solution of cholesteryl chloroformate (3.37 g, 7.5 mmol) in anhydrous dichloromethane (20 mL), triethylamine (TEA, 1.05 mL) was added under an argon atmosphere and stirred. Under ice-cooling, 6-(tert-butoxycarbonyl)amino-1-aminohexane (1.12 mL, 5 mmol) was added dropwise, and the mixture was stirred for 30 minutes under ice-cooling. After the temperature was raised to room temperature, the mixture was stirred overnight. The reaction mixture was washed with ultrapure water and saturated brine, dried over anhydrous magnesium sulfate, and the solvent was distilled off under reduced pressure. The resulting residue was purified by silica gel column chromatography (eluent: ethyl acetate:n-hexane = 1:4), and the fractions containing the target compound were combined, and the solvent was distilled off under reduced pressure.

[0166] (2) Step 1-2 The obtained residue was dissolved in ethyl acetate (40 mL), and a 4N hydrochloric acid / ethyl acetate solution (40 mL) was added thereto, followed by stirring at room temperature overnight. The resulting precipitate was collected by centrifugation. The obtained solid was washed four times with ethyl acetate, dried under reduced pressure, and purified to give cholesteryl 6-aminohexylcarbamate hydrochloride (Chol-C). ６ 1.2 g of the product (hydrochloride) was obtained. １ H-NMR spectrum (CS400 manufactured by JEOL, EtOH-d ６ ) is shown in Figure 1.

[0167] 2. Step 2: Preparation of tetrabutylammonium (TBA) salt of hyaluronic acid TBA salt of hyaluronic acid (HA-TBA) was prepared according to the following Step 2-1, followed by Step 2-2.

[0168] (1) Step 2-1: DOWEX (registered trademark) 50WX-8-400 (manufactured by Aldrich) was suspended in ultrapure water, and the resin was washed with ultrapure water approximately three times by decantation. A 40 wt % aqueous solution of tetrabutylammonium hydroxide (TBA-OH) (manufactured by Aldrich) was added in an amount of approximately 1.5 molar equivalents relative to the cation exchange capacity of the resin, and the mixture was stirred for 30 minutes. The excess TBA-OH solution was removed by decantation, and the resin was further washed with excess ultrapure water to obtain a TBA salt cation exchange resin.

[0169] (2) Step 2-2: Raw material hyaluronic acid sodium salt (HA-Na) with a molecular weight of 10,000 (10k) was dissolved in ultrapure water at a concentration of 15 mg / mL. A suspension of the cation exchange resin converted into a TBA salt in step 2-1 was added in an amount of 5 times the molar equivalent of the ion exchange capacity of the resin relative to the number of moles of HA unit (unit molecular weight 401.3). After stirring for 15 minutes, the mixture was filtered using a 0.45 μm filter, and the filtrate was freeze-dried to obtain the TBA salt of hyaluronic acid (HA-TBA) as a white solid. The product １ H-NMR spectrum (ECS400 manufactured by JEOL, EtOH-d ６ ) is shown in Figure 2.

[0170] 3. Step 3: A solution of HA-TBA prepared in Step 2-2 in anhydrous DMSO (10 mg / mL) was prepared. ６ At this time, the disaccharide repeating unit (HA unit) present in HA-TBA and Chol-C ６ The molar ratio of hydrochloride is HA unit / Chol-C ６ The ratio of the HA unit to the DMT-MM was adjusted to 100 / 44. Next, 4-(4,6-dimethoxy-1,3,5-triazin-2-yl)-4-methylmorpholinium chloride (DMT-MM) was added, and the mixture was stirred at room temperature overnight. At this time, the molar ratio of the HA unit to the DMT-MM was adjusted to 100 / 48.8. The reaction solution was dialyzed (Spectrapore 3, molecular weight cutoff (MWCO): 3,500) against a 0.3 M ammonium acetate / DMSO solution, a DMSO solution, a 0.15 M NaCl aqueous solution, and ultrapure water in that order. １ H-NMR spectrum (ECS400 manufactured by JEOL, EtOH-d ６ ) is shown in Figure 3. The peak (COCH) derived from the acetyl group of N-acetyl-D-glucosamine ３ , 1.6 ppm to 2.0 ppm, 3H), a peak derived from the methyl group in the cholesteryl group (CH ３ , 0.7 ppm, 3H) was confirmed.

[0171] [Production Example 2] (Hyaluronic acid derivative component HA-C ２ Preparation of aqueous solution of hyaluronic acid derivative component HA-C２ An aqueous solution of -Chol was prepared according to the following steps 1 to 3.

[0172] 1. Step 1: Synthesis of cholesteryl 2-aminoethylcarbamate hydrochloride Cholesteryl 2-aminoethylcarbamate hydrochloride (Chol-C ２ Cholesteryl 2-aminoethylcarbamate hydrochloride (Chol-C) was synthesized in the same manner as in Step 1 of Production Example 1, except that 2-(tert-butoxycarbonyl)amino-1-aminoethane (0.79 mL, 5 mmol) was used instead of 6-(tert-butoxycarbonyl)amino-1-aminohexane, and ethyl acetate:n-hexane=1:2 was used as the eluent for silica gel column chromatography. ２ 2.3 g of the compound (hydrochloride) was obtained.

[0173] 2. Step 2: Preparation of tetrabutylammonium (TBA) salt of hyaluronic acid The TBA salt of hyaluronic acid (HA-TBA) was prepared in the same manner as in Step 2 of Production Example 1.

[0174] 3. Step 3 Chol-C ６ Chol-C instead of hydrochloride ２ The hyaluronic acid derivative component HA-C was prepared in the same manner as in Step 3 of Production Example 1, except that the hydrochloride salt was used. ２ -An aqueous solution of Chol was obtained.

[0175] [Production Example 3] (Hyaluronic acid derivative component HA-C ８ Preparation of aqueous solution of hyaluronic acid derivative component HA-C ８ An aqueous solution of -Chol was prepared according to the following steps 1 to 3.

[0176] 1. Step 1: Synthesis of cholesteryl 8-aminooctylcarbamate hydrochloride Cholesteryl 8-aminooctylcarbamate hydrochloride (Chol-C ８ Hydrochloride) was synthesized according to the following steps 1-1 and then 1-2.

[0177] (1) Step 1-1: To a solution of 8-(t-butoxycarbonyl)amino-1-aminooctane (1.21 g, 5 mmol) in anhydrous dichloromethane (100 mL) and anhydrous toluene (200 mL), TEA (0.7 mL) was added under an argon atmosphere and stirred. Under ice-cooling, a solution of cholesteryl chloroformate (2.66 g, 6 mmol) in anhydrous dichloromethane was added dropwise, and the mixture was stirred for 30 minutes under ice-cooling, then warmed to room temperature and stirred overnight. The reaction mixture was concentrated under reduced pressure, and the resulting residue was purified by silica gel column chromatography (eluent: ethyl acetate:n-hexane = 1:4). The fractions containing the target compound were combined, and the solvent was evaporated under reduced pressure.

[0178] (2) Step 1-2 The resulting residue was dissolved in dichloromethane (1.5 mL), trifluoroacetic acid (1.5 mL) was added, and the mixture was stirred at room temperature for 2 hours. The solvent was evaporated under reduced pressure, and the resulting residue was purified by silica gel column chromatography (eluent: ethyl acetate:methanol:ammonia water=9:1:0.5). The target fractions were combined, and the solvent was evaporated under reduced pressure. 4N hydrochloric acid / dioxane was added to the resulting residue, and ethyl acetate was further added. The resulting solid was collected, washed with ethyl acetate, and dried under reduced pressure to give cholesteryl 8-aminooctylcarbamate (Chol-C8) hydrochloride (0.5 g).

[0179] 2. Step 2: Preparation of tetrabutylammonium (TBA) salt of hyaluronic acid The TBA salt of hyaluronic acid (HA-TBA) was prepared in the same manner as in Step 2 of Production Example 1.

[0180] 3. Step 3 Chol-C ６ Chol-C instead of hydrochloride ８ The hyaluronic acid derivative component HA-C was prepared in the same manner as in Step 3 of Production Example 1, except that the hydrochloride salt was used. ８ -An aqueous solution of Chol was obtained.

[0181] [Production Example 4] (Hyaluronic acid derivative component HA-C １２ Preparation of aqueous solution of hyaluronic acid derivative component HA-C １２ An aqueous solution of -Chol was prepared according to the following steps 1 to 3.

[0182] 1. Step 1: Synthesis of cholesteryl 12-aminododecylcarbamate hydrochloride Cholesteryl 12-aminododecylcarbamate hydrochloride (Chol-C １２ Cholesteryl 12-aminododecylcarbamate hydrochloride (Chol-C) was synthesized in the same manner as in Step 1 of Production Example 1, except that 12-(tert-butoxycarbonyl)amino-1-aminododecane (1.59 g, 5 mmol) was used instead of 6-(tert-butoxycarbonyl)amino-1-aminohexane. １２ Hydrochloride) 1.0 g was obtained.

[0183] 2. Step 2: Preparation of tetrabutylammonium (TBA) salt of hyaluronic acid The TBA salt of hyaluronic acid (HA-TBA) was prepared in the same manner as in Step 2 of Production Example 1.

[0184] 3. Step 3 Chol-C ６ Chol-C instead of hydrochloride １２ The hyaluronic acid derivative component HA-C was prepared in the same manner as in Step 3 of Production Example 1, except that the hydrochloride salt was used. １２ -An aqueous solution of Chol was obtained.

[0185] <Production of Carrier Compositions> [Examples 1 to 4] (Production of Carrier Compositions Ta1 to Ta4) Each of the carrier compositions was produced according to the following steps (A) to (C).

[0186] 1. Step (A): Adjustment of concentration of hyaluronic acid derivative component The aqueous solution of hyaluronic acid derivative component obtained in Preparation Example 1 is concentrated by centrifugal filtration unit (Vivaspin Turbo 15, MWCO=3,000, VS15T91, manufactured by Sartorius).The concentration of the hyaluronic acid derivative component contained in the aqueous solution (concentrate) of hyaluronic acid derivative component obtained is measured by size exclusion chromatography (SEC), and it is 10mg / mL.

[0187] 2. Step (B): Addition of cryoprotectant Sucrose was added as a cryoprotectant to the aqueous solution of the hyaluronic acid derivative component obtained in step (A) (concentration of the hyaluronic acid derivative component: 10 mg / mL) so that the mass ratio to the hyaluronic acid derivative component was 8.0 (Example 1), 4.0 (Example 2), 1.0 (Example 3), or 0.8 (Example 4), and the mixture was stirred to dissolve.

[0188] 3. Step (C): Freeze-drying The aqueous solution containing the hyaluronic acid derivative component and the cryoprotectant obtained in step (B) (concentration of the hyaluronic acid derivative component: 10 mg / mL, content of the cryoprotectant: 8, 10, 40, or 80 parts by mass / 10 parts by mass of the hyaluronic acid derivative component) was freeze-dried using the freeze-dryer shown below according to the following steps (C1) to (C4).

[0189] (Freeze dryer) Manufacturer: Takara AT&T Corporation Model: TF20-80TNNNS Shelf dimensions: 400W x 625D (mm) Number of shelves: 4 Shelf spacing: 100mm Shelf area: 0.25m ２ x 4 stages = 1.00m ２ Cold trap: Dehumidification capacity 20 liters Vacuum pump: Discharge capacity 300 liters / min

[0190] (1) Step (C1): Addition of Aqueous Solution The aqueous solution containing the hyaluronic acid derivative component and the cryoprotectant obtained in step (B) (concentration of the hyaluronic acid derivative component: 10 mg / mL, content of the cryoprotectant: 8, 10, 40, or 80 parts by mass / 10 parts by mass of the hyaluronic acid derivative component) was added to a SUS metal cup (bottom area: 22.9 cm ２ The amount of the aqueous solution containing the hyaluronic acid derivative component and the cryoprotectant was adjusted so that the thickness of the aqueous solution was 15 mm. The SUS metal cup containing the aqueous solution containing the hyaluronic acid derivative component and the cryoprotectant was placed on the sample shelf of the freeze-drying device.

[0191] In addition, to adjust the degree of vacuum inside the freeze-drying apparatus during primary drying, a total of 12.6 L of distilled water was poured into four aluminum trays, and the aluminum trays containing the distilled water were also placed on the sample shelves of the freeze-drying apparatus. At this time, the thickness of the distilled water was 15 mm.

[0192] (2) Step (C2): Pre-freezing First, all doors of the device are closed to form a closed system. The shelf temperature setting of the freeze-drying device is set to 20°C and maintained for 30 minutes. Then, the shelf temperature setting is lowered from 20°C to -40°C over one and a half hours, and then maintained for 10 hours to freeze the aqueous solution containing the hyaluronic acid derivative component and the cryoprotectant.

[0193] (3) Step (C3): Primary Drying The vacuum pump (full vacuum) and cold trap (set temperature -80°C) were started, and the shelf temperature setting was increased from -40°C to -20°C over 1 hour to initiate primary drying. The shelf temperature setting was then maintained at -20°C for 9 hours. Subsequently, the shelf temperature setting was increased from -20°C to -10°C over 1 hour, and the shelf temperature setting was maintained at -10°C for 91 hours, for a total of 102 hours from the start of primary drying. In step (C3), primary drying was achieved for at least 46 hours or more, starting 30 minutes after the start of step (C3), with a vacuum level in the apparatus of 5 Pa or more and less than 40 Pa. This was thought to be due to the fact that the amount of water vapor generated in the apparatus was increased by adjusting the amount of liquid (thickness of distilled water) in the apparatus in step (C1).

[0194] (4) Step (C4): Secondary drying The shelf temperature setting was increased from -10°C to 30°C over 1 hour, and then maintained at 30°C for 5 hours to perform secondary drying. After that, the device was stopped, the door of the freeze-dryer was opened, and the samples were removed to obtain each carrier composition.

[0195] [Examples 5 to 7] (Production of carrier compositions T-a5 to T-a7) In step (A) of Example 3, each carrier composition was obtained by the same method as in Example 3, except that the aqueous solution of the hyaluronic acid derivative component obtained in Production Example 1 was replaced with the aqueous solution of the hyaluronic acid derivative component obtained in Production Example 2 (Example 5), Production Example 3 (Example 6), or Production Example 4 (Example 7).

[0196] [Examples 8 to 9] (Production of carrier compositions T-a8 to T-a9) Each carrier composition was obtained in the same manner as in Example 1, except that in step (B) of Example 1, trehalose (Example 8) or glucose (Example 9) was used as the cryoprotectant instead of sucrose.

[0197] [Example 10] (Production of carrier composition T-a10) A carrier composition was obtained in the same manner as in Examples 1 to 4, except that in step (B) of Examples 1 to 4, sucrose was added as a cryoprotectant so that the mass ratio of the sucrose to the hyaluronic acid derivative component was 8.3.

[0198] [Examples 11-12] (Production of carrier compositions T-a11 to T-a12) The aqueous solution of hyaluronic acid derivative component and sucrose (concentration of hyaluronic acid derivative component: 10 mg / mL) obtained in step (B) of Example 3 was further diluted with water for injection (Otsuka distilled water for injection; manufactured by Otsuka Pharmaceutical Co., Ltd.) to adjust the concentration of hyaluronic acid derivative to 5.4 mg / mL (Example 11) or 1.4 mg / mL (Example 12), and carrier compositions were obtained in the same manner as in Example 3.

[0199] [Comparative Examples 1-2] (Production of Carrier Compositions T-b1-T-b2) In step (B) of Examples 1-4, each carrier composition was obtained in the same manner as in Examples 1-4, except that sucrose was added as a cryoprotectant so that the mass ratio to the hyaluronic acid derivative component was 0.5 (Comparative Example 1) or 0.1 (Comparative Example 2).

[0200] [Comparative Examples 3 to 5] (Production of Carrier Compositions T-b3 to T-b5) In step (A) of Comparative Example 2, each carrier composition was obtained by the same method as in Comparative Example 2, except that the aqueous solution of the hyaluronic acid derivative component obtained in Production Example 2 (Comparative Example 3), Production Example 3 (Comparative Example 4), or Production Example 4 (Comparative Example 5) was used instead of the aqueous solution of the hyaluronic acid derivative component obtained in Production Example 1.

[0201] [Comparative Examples 6 to 7] (Production of Carrier Compositions T-b6 to T-b7) Each carrier composition was obtained in the same manner as in Comparative Example 2, except that in step (B) of Comparative Example 2, trehalose (Comparative Example 6) or glucose (Comparative Example 7) was used as the cryoprotectant instead of sucrose.

[0202] Comparative Example 8 (Production of Carrier Composition T-b8) A carrier composition was obtained in the same manner as in Comparative Example 2, except that no cryoprotectant was added in step (B) of Comparative Example 2.

[0203] The physical properties and evaluation results of each carrier composition obtained in the Examples and Comparative Examples are shown in the following Table. Also, Figures 4 to 6 are graphs showing the relationship between the filtration resistance (Pa·s / m) and the amount of filtrate (mL) measured by a texture analyzer using each carrier composition obtained in the Examples and Comparative Examples.

[0204]

[0205]

[0206] From Table 1A, Table 1B, and Figures 4 to 6, it can be seen that in carrier compositions T-a1 to T-a12 (Examples 1 to 12) in which the content of sucrose, trehalose, or glucose was within a specific range relative to 10 parts by mass of the hyaluronic acid derivative component, the filtration resistance did not increase significantly and sterile filterability was good. This suggests that these carrier compositions can suppress filter clogging during sterile filtration during formulation. In addition, the undersieve mass fraction (%) of all carrier compositions T-a1 to T-a12 (Examples 1 to 12) after mortar grinding was 30% or more.

[0207] Among them, carrier compositions T-a1 to T-a11 (Examples 1 to 11) in which the concentration of the hyaluronic acid derivative component during freeze-drying in step (C) was within the range of 4 to 100 mg / mL exhibited particularly good sterile filtration properties. This suggests that these carrier compositions can particularly suppress filter clogging during sterile filtration during formulation.

[0208] Furthermore, in carrier compositions T-a1 to T-a8 and T-a11 to T-a12 (Examples 1 to 8, Examples 11 and 12) in which the content of sucrose or trehalose was 80 parts by mass or less per 10 parts by mass of the hyaluronic acid derivative component, the aqueous solution after reconstitution of the carrier composition was a hypotonic solution. Therefore, it is expected that the solution will be a hypotonic solution at the concentration used, and that it will be possible to further incorporate pharmaceutical additives in the formulation process.

[0209] On the other hand, in carrier compositions T-b1 to T-b8 (Comparative Examples 1 to 8) in which the content of sucrose, trehalose, or glucose was less than 6 parts by mass relative to 10 parts by mass of the hyaluronic acid derivative component, a significant increase in filtration resistance was observed, and sterile filtration was poor. This suggests that these carrier compositions are prone to filter clogging during sterile filtration during formulation. In addition, in carrier compositions T-b2 to T-b8 (Comparative Examples 2 to 8) in which the content of sucrose, trehalose, or glucose was less than 5 parts by mass relative to 10 parts by mass of the hyaluronic acid derivative component, the undersieve mass fraction (%) after mortar grinding was all less than 30%.

[0210] <Production of aqueous solution of hyaluronic acid derivative component: b> [Production Example 1-b] (hyaluronic acid derivative component HA-C ６ Preparation of aqueous solution of hyaluronic acid derivative component HA-C ６ An aqueous solution of -Chol was prepared according to the following steps 1 to 3.

[0211] 1. Step 1: Synthesis of cholesteryl 6-aminohexylcarbamate hydrochloride Cholesteryl 6-aminohexylcarbamate hydrochloride (Chol-C ６ Hydrochloride) was synthesized in the same manner as described in Step 1 of Preparation 1.

[0212] 2. Step 2: Preparation of tetrabutylammonium (TBA) salt of hyaluronic acid TBA salt of hyaluronic acid (HA-TBA) was prepared in the same manner as described in Step 2 of Preparation Example 1.

[0213] 3. Step 3: A solution of HA-TBA prepared in Step 2-2 in anhydrous DMSO (10 mg / mL) was prepared.６ At this time, the disaccharide repeating unit (HA unit) present in HA-TBA and Chol-C ６ The molar ratio of hydrochloride is HA unit / Chol-C ６ The ratio of the HA unit to the DMT-MM was adjusted to 100 / 31. Next, 4-(4,6-dimethoxy-1,3,5-triazin-2-yl)-4-methylmorpholinium chloride (DMT-MM) was added and the mixture was stirred at room temperature overnight. At this time, the molar ratio of the HA unit to the DMT-MM was adjusted to 100 / 40.4. The reaction solution was dialyzed (Spectrapore 3, molecular weight cutoff (MWCO): 3,500) against a 0.3 M ammonium acetate / DMSO solution, a DMSO solution, a 0.15 M NaCl aqueous solution, and ultrapure water in that order. １ H-NMR spectrum (ECS400 manufactured by JEOL, EtOH-d ６ ) is shown in Figure 7. The peak (COCH) derived from the acetyl group of N-acetyl-D-glucosamine ３ , 1.6 ppm to 2.0 ppm, 3H), a peak derived from the methyl group in the cholesteryl group (CH ３ , 0.7 ppm, 3H) was confirmed.

[0214] [Production Example 2-b] (Hyaluronic acid derivative component HA-C ２ Preparation of aqueous solution of hyaluronic acid derivative component HA-C ２ An aqueous solution of -Chol was prepared according to the following steps 1 to 3.

[0215] 1. Step 1: Synthesis of cholesteryl 2-aminoethylcarbamate hydrochloride Cholesteryl 2-aminoethylcarbamate hydrochloride (Chol-C ２ Hydrochloride) was synthesized in the same manner as described in Step 1 of Preparation Example 2.

[0216] 2. Step 2: Preparation of tetrabutylammonium (TBA) salt of hyaluronic acid TBA salt of hyaluronic acid (HA-TBA) was prepared in the same manner as described in Step 2 of Production Example 2.

[0217] 3. Step 3 Chol-C ６ Chol-C instead of hydrochloride ２The hyaluronic acid derivative component HA-C was obtained in the same manner as in step 3 of Production Example 1-b, except that the hydrochloride salt was used. ２ -An aqueous solution of Chol was obtained.

[0218] [Production Example 3-b] (Hyaluronic acid derivative component HA-C ８ Preparation of aqueous solution of hyaluronic acid derivative component HA-C ８ An aqueous solution of -Chol was prepared according to the following steps 1 to 3.

[0219] 1. Step 1: Synthesis of cholesteryl 8-aminooctylcarbamate hydrochloride Cholesteryl 8-aminooctylcarbamate hydrochloride (Chol-C ８ Hydrochloride) was synthesized in the same manner as described in Step 1 of Preparation Example 3.

[0220] 2. Step 2: Preparation of tetrabutylammonium (TBA) salt of hyaluronic acid TBA salt of hyaluronic acid (HA-TBA) was prepared in the same manner as in Step 2 of Production Example 3.

[0221] 3. Step 3 Chol-C ６ Chol-C instead of hydrochloride ８ The hyaluronic acid derivative component HA-C was obtained in the same manner as in step 3 of Production Example 1-b, except that the hydrochloride salt was used. ８ -An aqueous solution of Chol was obtained.

[0222] [Production Example 4-b] (Hyaluronic acid derivative component HA-C １２ Preparation of aqueous solution of hyaluronic acid derivative component HA-C １２ An aqueous solution of -Chol was prepared according to the following steps 1 to 3.

[0223] 1. Step 1: Synthesis of cholesteryl 12-aminododecylcarbamate hydrochloride Cholesteryl 12-aminododecylcarbamate hydrochloride (Chol-C １２ Hydrochloride) was synthesized in the same manner as described in Step 1 of Production Example 4.

[0224] 2. Step 2: Preparation of tetrabutylammonium (TBA) salt of hyaluronic acid TBA salt of hyaluronic acid (HA-TBA) was prepared in the same manner as in Step 2 of Production Example 4.

[0225] 3. Step 3 Chol-C ６ Chol-C instead of hydrochloride １２ The hyaluronic acid derivative component HA-C was obtained in the same manner as in step 3 of Production Example 1-b, except that the hydrochloride salt was used. １２ -An aqueous solution of Chol was obtained.

[0226] <Production of Carrier Composition: b> [Examples 1-b to 3-b] (Production of Carrier Compositions Tb-a1 to Tb-a3) Each of the carrier compositions was produced according to the following steps (A) to (C).

[0227] 1. Step (A): Adjustment of the concentration of the hyaluronic acid derivative component The aqueous solution of the hyaluronic acid derivative component obtained in Production Example 1-b was concentrated using a centrifugal filtration unit (Vivaspin Turbo 15, MWCO = 3,000, VS15T91, manufactured by Sartorius). The concentration of the hyaluronic acid derivative component contained in the resulting aqueous solution of the hyaluronic acid derivative component (concentrate) was measured by size exclusion chromatography (SEC) and found to be 11.4 mg / mL.

[0228] 2. Step (B): Addition of cryoprotectant Sucrose was added as a cryoprotectant to the aqueous solution of the hyaluronic acid derivative component obtained in step (A) (concentration of the hyaluronic acid derivative component: 11.4 mg / mL) so that the mass ratio to the hyaluronic acid derivative component was 8.0 (Example 1-b), 4.0 (Example 2-b), or 1.0 (Example 3-b), and the mixture was stirred to dissolve.

[0229] 3. Step (C): Freeze-drying The aqueous solution containing the hyaluronic acid derivative component and the cryoprotectant obtained in step (B) (the concentration of the hyaluronic acid derivative component: 11.4 mg / mL, the content of the cryoprotectant: 10, 40, or 80 parts by mass / 10 parts by mass of the hyaluronic acid derivative component) was freeze-dried in the same manner as in the method described in Examples 1 to 4 above, to obtain each carrier composition.

[0230] [Examples 4-b to 6-b] (Production of carrier compositions Tb-a4 to Tb-a6) In step (A) of Example 3-b, each carrier composition was obtained by the same method as in Example 3-b, except that in place of the aqueous solution of the hyaluronic acid derivative component obtained in Production Example 1-b, the aqueous solution of the hyaluronic acid derivative component obtained in Production Example 2-b (Example 4-b), Production Example 3-b (Example 5-b), or Production Example 4-b (Example 6-b) was used.

[0231] [Examples 7-b to 9-b] (Production of carrier compositions Tb-a7 to Tb-a9) Each carrier composition was obtained in the same manner as in Example 1-b, except that in step (B) of Example 1-b, trehalose (Example 7-b), glucose (Example 8-b), or maltose (Example 9-b) was used as the cryoprotectant instead of sucrose.

[0232] [Example 10-b] (Production of carrier composition Tb-a10) A carrier composition was obtained in the same manner as in Examples 1-b to 3-b, except that in step (B) of Examples 1-b to 3-b, sucrose was added as a cryoprotectant so that the mass ratio of the sucrose to the hyaluronic acid derivative component was 8.3.

[0233] [Examples 11-b to 12-b] (Production of carrier compositions Tb-a11 to Tb-a12) The aqueous solution of the hyaluronic acid derivative component and sucrose (concentration of the hyaluronic acid derivative component: 10 mg / mL) obtained in step (B) of Example 3-b was further diluted with water for injection (Otsuka distilled water for injection; manufactured by Otsuka Pharmaceutical Co., Ltd.) to adjust the concentration of the hyaluronic acid derivative to 5.4 mg / mL (Example 11-b) or 1.4 mg / mL (Example 12-b), except that a carrier composition was obtained in the same manner as in Example 3-b.

[0234] [Comparative Examples 1-b to 2-b] (Production of Carrier Compositions Tb-b1 to Tb-b2) In step (B) of Examples 1-b to 3-b, each carrier composition was obtained in the same manner as in Examples 1-b to 3-b, except that sucrose was added as a cryoprotectant so that the mass ratio to the hyaluronic acid derivative component was 0.5 (Comparative Example 1-b) or 0.1 (Comparative Example 2-b).

[0235] [Comparative Examples 3-b to 5-b] (Production of Carrier Compositions Tb-b3 to Tb-b5) In step (A) of Comparative Example 2-b, each carrier composition was obtained by the same method as in Comparative Example 2-b, except that the aqueous solution of the hyaluronic acid derivative component obtained in Production Example 2-b (Comparative Example 3-b), Production Example 3-b (Comparative Example 4-b), or Production Example 4-b (Comparative Example 5-b) was used instead of the aqueous solution of the hyaluronic acid derivative component obtained in Production Example 1-b.

[0236] [Comparative Examples 6-b to 8-b] (Production of Carrier Compositions Tb-b6 to Tb-b8) Each carrier composition was obtained in the same manner as in Comparative Example 2-b, except that in step (B) of Comparative Example 2-b, trehalose (Comparative Example 6-b), glucose (Comparative Example 7-b), or maltose (Comparative Example 8-b) was used as the cryoprotectant instead of sucrose.

[0237] [Comparative Example 9-b] (Production of Carrier Composition Tb-b9) A carrier composition was obtained in the same manner as in Comparative Example 2-b, except that no cryoprotectant was added in step (B) of Comparative Example 2-b.

[0238] The physical properties and evaluation results of each carrier composition obtained in the Examples and Comparative Examples are shown in the following table. Also, Figures 8 to 10 are graphs showing the relationship between the filtration resistance (Pa·s / m) and the amount of filtrate (mL) measured by a texture analyzer using each carrier composition obtained in the Examples and Comparative Examples.

[0239]

[0240]

[0241] From Tables 2A, 2B, and Figures 8 to 10, it can be seen that in carrier compositions Tb-a1 to Tb-a12 (Examples 1-b to 12-b), in which the content of sucrose, trehalose, glucose, or maltose was within a specific range relative to 10 parts by mass of the hyaluronic acid derivative component, the filtration resistance did not increase significantly and the sterile filterability was good. This suggests that these carrier compositions can suppress filter clogging during sterile filtration during formulation. In addition, the undersieve mass fraction (%) of all carrier compositions Tb-a1 to Tb-a12 (Examples 1-b to 12-b) after mortar grinding was 30% or more.

[0242] Among them, carrier compositions Tb-a1 to Tb-a11 (Examples 1-b to 11-b), in which the concentration of the hyaluronic acid derivative component during freeze-drying in step (C) was within the range of 4 to 100 mg / mL, exhibited particularly good sterile filtration properties. This suggests that these carrier compositions can particularly suppress filter clogging during sterile filtration during formulation.

[0243] Furthermore, in carrier compositions Tb-a1 to Tb-a7, Tb-a9, and Tb-a11 to Tb-a12 (Examples 1-b to 7-b, Example 9-b, and Example 11-b to 12-b), in which the content of sucrose, trehalose, or maltose was 80 parts by mass or less per 10 parts by mass of the hyaluronic acid derivative component, the aqueous solution after reconstitution of the carrier composition was a hypotonic solution. Therefore, it is expected that the solution will be a hypotonic solution at the usage concentration, and that it will be possible to further incorporate pharmaceutical additives in the formulation process.

[0244] On the other hand, in carrier compositions Tb-b1 to Tb-b9 (Comparative Examples 1-b to 9-b), in which the content of sucrose, trehalose, glucose, or maltose was less than 6 parts by mass relative to 10 parts by mass of the hyaluronic acid derivative component, a significant increase in filtration resistance was observed, and sterile filtration was poor. This suggests that these carrier compositions are prone to filter clogging during sterile filtration during formulation. In addition, in carrier compositions Tb-b2 to Tb-b9 (Comparative Examples 2-b to 9-b), in which the content of sucrose, trehalose, glucose, or maltose was less than 5 parts by mass relative to 10 parts by mass of the hyaluronic acid derivative component, the undersieve mass fraction (%) after mortar grinding was all less than 30%.

[0245] <Production of aqueous solution of hyaluronic acid derivative component: c> [Production Example 1-c] (hyaluronic acid derivative component HA-C ６ Preparation of aqueous solution of hyaluronic acid derivative component HA-C ６ An aqueous solution of -Chol was prepared according to the following steps 1 to 3.

[0246] 1. Step 1: Synthesis of cholesteryl 6-aminohexylcarbamate hydrochloride Cholesteryl 6-aminohexylcarbamate hydrochloride (Chol-C ６ Hydrochloride) was synthesized in the same manner as described in Step 1 of Preparation Example 1.

[0247] 2. Step 2: Preparation of tetrabutylammonium (TBA) salt of hyaluronic acid TBA salt of hyaluronic acid (HA-TBA) was prepared according to the following Step 2-1, followed by Step 2-2.

[0248] (1) Step 2-1: DOWEX (registered trademark) 50WX-8-400 (manufactured by Aldrich) was suspended in ultrapure water, and the resin was washed with ultrapure water approximately three times by decantation. A 40 wt % aqueous solution of tetrabutylammonium hydroxide (TBA-OH) (manufactured by Aldrich) was added in an amount of approximately 1.5 molar equivalents relative to the cation exchange capacity of the resin, and the mixture was stirred for 30 minutes. The excess TBA-OH solution was removed by decantation, and the resin was further washed with excess ultrapure water to obtain a TBA salt cation exchange resin.

[0249] (2) Step 2-2: Raw material hyaluronic acid sodium salt (HA-Na) with a molecular weight of 35,000 (35k) was dissolved in ultrapure water at a concentration of 15 mg / mL. A suspension of the cation exchange resin converted into a TBA salt in step 2-1 was added in an amount of 5 times the molar equivalent of the ion exchange capacity of the resin relative to the number of moles of HA unit (unit molecular weight 401.3). After stirring for 15 minutes, the mixture was filtered using a 0.45 μm filter, and the filtrate was freeze-dried to obtain the TBA salt of hyaluronic acid (HA-TBA) as a white solid. The product １ H-NMR spectrum (ECS400 manufactured by JEOL, EtOH-d ６ ) is shown in FIG.

[0250] 3. Step 3: A solution of HA-TBA prepared in Step 2-2 in anhydrous DMSO (10 mg / mL) was prepared. ６ At this time, the disaccharide repeating unit (HA unit) present in HA-TBA and Chol-C ６ The molar ratio of hydrochloride is HA unit / Chol-C ６ The ratio of the HA unit to the DMT-MM was adjusted to 100 / 20. Next, 4-(4,6-dimethoxy-1,3,5-triazin-2-yl)-4-methylmorpholinium chloride (DMT-MM) was added, and the mixture was stirred at room temperature overnight. At this time, the molar ratio of the HA unit to the DMT-MM was adjusted to 100 / 22.0. The reaction solution was dialyzed (Spectrapore 3, molecular weight cutoff (MWCO): 3,500) against a 0.3 M ammonium acetate / DMSO solution, a DMSO solution, a 0.15 M NaCl aqueous solution, and ultrapure water in that order. １ H-NMR spectrum (ECS400 manufactured by JEOL, EtOH-d ６ ) is shown in Figure 12. The peak (COCH) derived from the acetyl group of N-acetyl-D-glucosamine ３ , 1.6 ppm to 2.0 ppm, 3H), a peak derived from the methyl group in the cholesteryl group (CH ３ , 0.7 ppm, 3H) was confirmed.

[0251] [Production Example 2-c] (Hyaluronic acid derivative component HA-C ２ Preparation of aqueous solution of hyaluronic acid derivative component HA-C２ An aqueous solution of -Chol was prepared according to the following steps 1 to 3.

[0252] 1. Step 1: Synthesis of cholesteryl 2-aminoethylcarbamate hydrochloride Cholesteryl 2-aminoethylcarbamate hydrochloride (Chol-C ２ Hydrochloride) was synthesized in the same manner as described in Step 1 of Production Example 2.

[0253] 2. Step 2: Preparation of tetrabutylammonium (TBA) salt of hyaluronic acid TBA salt of hyaluronic acid (HA-TBA) was prepared in the same manner as in Step 2 of Production Example 3.

[0254] 3. Step 3 Chol-C ６ Chol-C instead of hydrochloride ２ The hyaluronic acid derivative component HA-C was obtained in the same manner as in step 3 of Production Example 1-c, except that the hydrochloride salt was used. ２ -An aqueous solution of Chol was obtained.

[0255] [Production Example 3-c] (Hyaluronic acid derivative component HA-C ８ Preparation of aqueous solution of hyaluronic acid derivative component HA-C ８ An aqueous solution of -Chol was prepared according to the following steps 1 to 3.

[0256] 1. Step 1: Synthesis of cholesteryl 8-aminooctylcarbamate hydrochloride Cholesteryl 8-aminooctylcarbamate hydrochloride (Chol-C ８ Hydrochloride) was prepared in a manner similar to that described in Step 1 of Preparation 3.

[0257] 2. Step 2: Preparation of tetrabutylammonium (TBA) salt of hyaluronic acid TBA salt of hyaluronic acid (HA-TBA) was prepared in the same manner as in the step of Production Example 3.

[0258] 3. Step 3 Chol-C ６ Chol-C instead of hydrochloride ８ The hyaluronic acid derivative component HA-C was obtained in the same manner as in step 3 of Production Example 1-c, except that the hydrochloride salt was used. ８ -An aqueous solution of Chol was obtained.

[0259] [Production Example 4-c] (Hyaluronic acid derivative component HA-C １２ Preparation of aqueous solution of hyaluronic acid derivative component HA-C １２ An aqueous solution of -Chol was prepared according to the following steps 1 to 3.

[0260] 1. Step 1: Synthesis of cholesteryl 12-aminododecylcarbamate hydrochloride Cholesteryl 12-aminododecylcarbamate hydrochloride (Chol-C １２ Hydrochloride) was synthesized in the same manner as described in Step 1 of Production Example 4.

[0261] 2. Step 2: Preparation of tetrabutylammonium (TBA) salt of hyaluronic acid TBA salt of hyaluronic acid (HA-TBA) was prepared in the same manner as in Step 2 of Production Example 4.

[0262] 3. Step 3 Chol-C ６ Chol-C instead of hydrochloride １２ The hyaluronic acid derivative component HA-C was obtained in the same manner as in step 3 of Production Example 1-c, except that the hydrochloride salt was used. １２ -An aqueous solution of Chol was obtained.

[0263] <Production of Carrier Compositions> [Examples 1-c to 3-c] (Production of Carrier Compositions Tc-a1 to Tc-a3) Each carrier composition was produced according to the following steps (A) to (C).

[0264] 1. Step (A): Adjustment of the concentration of the hyaluronic acid derivative component The aqueous solution of the hyaluronic acid derivative component obtained in Production Example 1-c was concentrated using a centrifugal filtration unit (Vivaspin Turbo 15, MWCO = 3,000, VS15T91, manufactured by Sartorius). The concentration of the hyaluronic acid derivative component contained in the resulting aqueous solution of the hyaluronic acid derivative component (concentrate) was measured by size exclusion chromatography (SEC) and found to be 5.6 mg / mL.

[0265] 2. Step (B): Addition of cryoprotectant Sucrose was added as a cryoprotectant to the aqueous solution of the hyaluronic acid derivative component obtained in step (A) (concentration of the hyaluronic acid derivative component: 5.6 mg / mL) so that the mass ratio to the hyaluronic acid derivative component was 18.5 (Example 1), 8.0 (Example 2), or 3.0 (Example 3), and the mixture was stirred to dissolve.

[0266] 3. Step (C): Freeze-drying The aqueous solution containing the hyaluronic acid derivative component and the cryoprotectant obtained in step (B) (the concentration of the hyaluronic acid derivative component: 5.6 mg / mL, the content of the cryoprotectant: 30, 80, or 185 parts by mass / 10 parts by mass of the hyaluronic acid derivative component) was freeze-dried in the same manner as in the method described in the above-mentioned [Examples 1 to 4], thereby obtaining each carrier composition.

[0267] [Examples 4-c to 6-c] (Production of carrier compositions Tc-a4 to Tc-a6) In step (A) of Example 3-c, each carrier composition was obtained by the same method as in Example 3-c, except that the aqueous solution of the hyaluronic acid derivative component obtained in Production Example 2-c (Example 4-c), Production Example 3-c (Example 5-c), or Production Example 4-c (Example 6-c) was used instead of the aqueous solution of the hyaluronic acid derivative component obtained in Production Example 1-c.

[0268] [Examples 7-c to 8-c] (Production of carrier compositions Tc-a7 to Tc-a8) Each carrier composition was obtained by the same method as in Example 3-c, except that in step (B) of Example 3-c, maltose (Example 7-c) or sorbitol (Example 8-c) was used as the cryoprotectant instead of sucrose.

[0269] [Example 9-c] A carrier composition was obtained in the same manner as in Examples 1-c to 3-c, except that in step (B) of Examples 1-c to 3-c, sucrose was added as a cryoprotectant so that the mass ratio of the sucrose to the hyaluronic acid derivative component was 20.0.

[0270] [Comparative Examples 1-c to 2-c] (Production of Carrier Compositions Tc-b1 to Tc-b2) In step (B) of Examples 1-c to 3-c, each carrier composition was obtained by the same method as in Examples 1-c to 3-c, except that sucrose was added as a cryoprotectant so that the mass ratio to the hyaluronic acid derivative component was 2.5 (Comparative Example 1-c) or 1.0 (Comparative Example 2-c).

[0271] [Comparative Examples 3-c to 5-c] (Production of Carrier Compositions Tc-b3 to Tc-b5) In step (A) of Comparative Example 2-c, each carrier composition was obtained by the same method as in Comparative Example 2-c, except that the aqueous solution of the hyaluronic acid derivative component obtained in Production Example 2-c (Comparative Example 3-c), Production Example 3-c (Comparative Example 4-c), or Production Example 4-c (Comparative Example 5-c) was used instead of the aqueous solution of the hyaluronic acid derivative component obtained in Production Example 1-c.

[0272] [Comparative Examples 6-c to 7-c] (Production of Carrier Compositions Tc-b6 to Tc-b7) Each carrier composition was obtained by the same method as in Comparative Example 2-c, except that in step (B) of Comparative Example 2-c, maltose (Comparative Example 6) or sorbitol (Comparative Example 7-c) was used as the cryoprotectant instead of sucrose.

[0273] [Comparative Example 8-c] (Production of carrier composition Tc-b8) A carrier composition was obtained in the same manner as in Comparative Example 2-c, except that in step (B) of Comparative Example 2-c, sucrose was added as a cryoprotectant so that the mass ratio to the hyaluronic acid derivative component was 0.5.

[0274] [Comparative Example 9-c] (Production of Carrier Composition Tc-b9) A carrier composition was obtained in the same manner as in Comparative Example 2-c, except that no cryoprotectant was added in step (B) of Comparative Example 2-c.

[0275] [Comparative Example 10-c] (Production of Carrier Composition Tc-b10) A carrier composition was obtained in the same manner as in Comparative Example 2-c, except that in step (B) of Comparative Example 2-c, sucrose was added as a cryoprotectant so that the mass ratio relative to the hyaluronic acid derivative component was 8.3, and sodium chloride was added so that the mass ratio relative to the hyaluronic acid derivative component was 0.18.

[0276] The physical properties and evaluation results of each carrier composition obtained in the Examples and Comparative Examples are shown in the following Table. Also, Figures 13 to 15 are graphs showing the relationship between the filtration resistance (Pa·s / m) and the amount of filtrate (mL) measured by a texture analyzer using each carrier composition obtained in the Examples and Comparative Examples.

[0277]

[0278]

[0279] From Table 3A, Table 3B, and Figures 13 to 15, the carrier compositions Tc-a1 to Tc-a9 (Examples 1-c to 9-c), in which the content of sucrose, maltose, or sorbitol is within a specific range relative to 10 parts by mass of the hyaluronic acid derivative component, and the salt content is within a specific range relative to 10 parts by mass of the hyaluronic acid derivative component, did not show a significant increase in filtration resistance and had good sterile filterability. This suggests that these carrier compositions can suppress filter clogging during sterile filtration during formulation. In addition, the carrier compositions Tc-a1 to Tc-a9 (Examples 1-c to 9-c) all had a mortar-pulverized undersieve mass fraction (%) of 30% or more.

[0280] In addition, in carrier compositions Tc-a1 to Tc-a8 (Examples 1-c to 8-c) in which the content of sucrose, maltose, or sorbitol was 185 parts by mass or less per 10 parts by mass of the hyaluronic acid derivative component, the aqueous solution after reconstitution of the carrier composition was a hypotonic solution. Therefore, it is expected that the solution will be a hypotonic solution at the use concentration, and that it will be possible to further incorporate pharmaceutical additives in the formulation process.

[0281] On the other hand, in carrier compositions Tc-b1 to Tc-b9 (Comparative Examples 1-c to 9-c), in which the content of sucrose, maltose, or sorbitol was less than 28 parts by mass relative to 10 parts by mass of the hyaluronic acid derivative component, a significant increase in filtration resistance was observed, and sterile filtration was poor. This suggests that these carrier compositions are prone to filter clogging during sterile filtration during formulation.

[0282] The carrier composition of this embodiment can prevent clogging of the filter during sterile filtration in the formulation process.

Claims

1. A carrier composition used in a formulation prepared by mixing an active ingredient in a form dissolved in a solvent and then sterilizing and filtering, the carrier composition comprising: a hyaluronic acid derivative component containing hyaluronic acid and a hyaluronic acid derivative having a steryl group introduced therein; and a cryoprotectant; the hyaluronic acid derivative has a repeating unit represented by the following general formula (I), the introduction rate of the steryl group relative to the repeating units of disaccharides derived from the hyaluronic acid and the hyaluronic acid derivative is 25% or more and 60% or less, the content of the cryoprotectant is 6 parts by mass or more and 1750 parts by mass or less per 10 parts by mass of the hyaluronic acid derivative component, and the carrier composition is a freeze-dried product. (In the formula, R １ , R ２ , R ３ , and R ４ are each independently a hydrogen atom, C １－６ Alkyl, formyl and C １－６ alkylcarbonyl; Z represents a direct bond or a peptide linker consisting of any amino acid residues of 2 to 30; X １ is represented by the following formula: ｂ -R, -NR ｂ -COO-R, -NR ｂ -CO-R, -NR ｂ -CO-NR ｃ -R, -COO-R, -O-COO-R, -SR, -CO-Y ａ -SR, -O-CO-Y ｂ -SR, -NR ｂ -CO-Y ｂ -SR, and -S-SR, a group selected from the group consisting of groups represented by R ａ , R ｂ and R ｃ are each independently a hydrogen atom, C １－２０ Alkyl, Amino C ２－２０ Alkyl and hydroxy C ２－２０ alkyl, where the alkyl portion of the group is selected from the group consisting of —O— and —NR ｆ - may be inserted with a group selected from the group consisting of; R ｆ is a hydrogen atom, C １－１２ Alkyl, Amino C ２－１２ Alkyl and hydroxy C ２－１２ alkyl, the alkyl portion of which may be interrupted by a group selected from the group consisting of -O- and -NH-; R is a steryl group; Y is C ２－３０ Alkylene, or -(CH ２ CH ２ O) ｍ -CH ２ CH ２ -, where the alkylene is -O-, -NR ｇ - and -S-S- may be inserted; R ｇ is a hydrogen atom, C １－２０ Alkyl, Amino C ２－２０ Alkyl and hydroxy C ２－２０ alkyl, the alkyl portion of which may be inserted with a group selected from the group consisting of —O— and —NH—; Y ａ is C １－５ is alkylene; Y ｂ is C ２－８ Alkylene or C ２－８ alkenylene; and m is an integer of 1 or more and 100 or less.

2. The carrier composition according to claim 1, wherein the content of the cryoprotectant is 6 to 80 parts by mass per 10 parts by mass of the hyaluronic acid derivative component.

3. The carrier composition according to claim 1 or 2, wherein the molecular weight of the cryoprotectant is 200 or more.

4. The carrier composition according to claim 1 or 2, wherein the cryoprotectant is a sugar or a sugar alcohol.

5. The carrier composition of claim 4, wherein the sugar is one or more selected from the group consisting of sucrose, trehalose, and glucose.

6. The carrier composition of claim 4, wherein the sugar alcohol is sorbitol.

7. The carrier composition of claim 1 or 2, wherein R is a cholesteryl group or a phytosteryl group.

8. The carrier composition of claim 1 or 2, wherein R is a cholesteryl group.

9. A carrier composition according to claim 1 or 2, wherein the molecular weight of the hyaluronic acid derivative component is 1,000 or more but less than 20,000.

10. A carrier composition described in claim 1 or 2, in which, after the carrier composition is crushed in a mortar, the proportion of the material passing through a sieve with a mesh size of 2.0 mm according to the dry sieving test method specified in JIS K 0069:1992 is 30 mass% or more.

11. A carrier composition used in a formulation obtained by mixing the active ingredient in a form dissolved in a solvent and then sterilizing and filtering, the carrier composition comprising: a hyaluronic acid derivative component containing hyaluronic acid and a hyaluronic acid derivative into which a steryl group has been introduced; and a cryoprotectant; the hyaluronic acid derivative has a repeating unit represented by the following general formula (I); the introduction rate of the steryl group relative to the repeating units of disaccharides derived from the hyaluronic acid and the hyaluronic acid derivative is 0.1% or more and less than 25%; the content of the cryoprotectant is 28 parts by mass or more and 1750 parts by mass or less per 10 parts by mass of the hyaluronic acid derivative component; and the salt content is 0.001 parts by mass or more and less than 0.500 parts by mass per 10 parts by mass of the hyaluronic acid derivative component; and the carrier composition is a freeze-dried product. (In the formula, R １ , R ２ , R ３ , and R ４ are each independently a hydrogen atom, C １－６ Alkyl, formyl and C １－６ alkylcarbonyl; Z represents a direct bond or a peptide linker consisting of any amino acid residues of 2 to 30; X １ is represented by the following formula: ｂ -R, -NR ｂ -COO-R, -NR ｂ -CO-R, -NR ｂ -CO-NR ｃ -R, -COO-R, -O-COO-R, -SR, -CO-Y ａ -SR, -O-CO-Y ｂ -SR, -NR ｂ -CO-Y ｂ -SR, and -S-SR, a group selected from the group consisting of groups represented by R ａ , R ｂ and R ｃ are each independently a hydrogen atom, C １－２０ Alkyl, Amino C ２－２０ Alkyl and hydroxy C ２－２０ alkyl, where the alkyl portion of the group is selected from the group consisting of —O— and —NR ｆ - may be inserted with a group selected from the group consisting of; R ｆ is a hydrogen atom, C １－１２ Alkyl, Amino C ２－１２ Alkyl and hydroxy C ２－１２ alkyl, the alkyl portion of which may be interrupted by a group selected from the group consisting of -O- and -NH-; R is a steryl group; Y is C ２－３０ Alkylene, or -(CH ２ CH ２ O) ｍ -CH ２ CH ２ -, where the alkylene is -O-, -NR ｇ - and -S-S- may be inserted; R ｇ is a hydrogen atom, C １－２０ Alkyl, Amino C ２－２０ Alkyl and hydroxy C ２－２０ alkyl, the alkyl portion of which may be inserted with a group selected from the group consisting of —O— and —NH—; Y ａ is C １－５ is alkylene; Y ｂ is C ２－８ Alkylene or C ２－８ alkenylene; and m is an integer of 1 or more and 100 or less.

12. The carrier composition described in claim 11, wherein the content of the cryoprotectant is 28 parts by mass or more and 185 parts by mass or less per 10 parts by mass of the hyaluronic acid derivative component.

13. The carrier composition according to claim 11 or 12, wherein the molecular weight of the cryoprotectant is 200 or more.

14. The carrier composition of claim 11 or 12, wherein the cryoprotectant is a sugar or sugar alcohol.

15. The carrier composition of claim 14, wherein the sugar is sucrose or maltose.

16. The carrier composition of claim 14, wherein the sugar alcohol is sorbitol.

17. The carrier composition of claim 11 or 12, wherein R is a cholesteryl group or a phytosteryl group.

18. The carrier composition of claim 11 or 12, wherein R is a cholesteryl group.

19. The carrier composition according to claim 11 or 12, wherein the molecular weight of the hyaluronic acid derivative component is 1,000 or more but less than 1,000,000.

20. A carrier composition described in claim 11 or 12, in which, after the carrier composition is crushed in a mortar, the proportion of the material passing through a sieve with a mesh size of 2.0 mm according to the dry sieving test method specified in JIS K 0069:1992 is 30 mass% or more.

21. A method for producing a carrier composition, comprising: a step (A) of adjusting the concentration of a hyaluronic acid derivative component in an aqueous solution to be 4 mg / mL or more and 100 mg / mL or less; a step (B) of adding a cryoprotectant to the aqueous solution after step (A); and a step (C) of freeze-drying the aqueous solution containing the hyaluronic acid derivative component and the cryoprotectant.

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