Hyaluronic acid derivative pharmaceutical composition for oral absorption

Abstract

To provide a hyaluronic acid derivative pharmaceutical composition for oral absorption excellent in drug absorbability.SOLUTION: An orally absorbable hyaluronic acid derivative pharmaceutical composition comprising (A) a hyaluronic acid derivative having an introduced steryl group and (B) an active ingredient, wherein the content of the (B) active ingredient relative to 100 parts by mass of the (A) hyaluronic acid derivative is 0.1 parts by mass or more and less than 10,000 parts by mass, and the average molecular weight of the (A) hyaluronic acid derivative is 6,000 or more and 20,000 or less.SELECTED DRAWING: None

Description

[Technical Field]

[0001] The present invention relates to a pharmaceutical composition that uses a hyaluronic acid derivative as a carrier and has improved oral absorbability. [Background technology]

[0002] In recent years, biopharmaceuticals, which are pharmaceuticals that use proteins, peptides, or nucleic acids as active ingredients, have been put into practical use, and their number is increasing year by year. Biopharmaceuticals can fulfill unmet medical needs that could not be met by conventional small molecule drugs.

[0003] However, biopharmaceuticals have the drawbacks of being poorly absorbed through the digestive tract or mucous membranes, being unstable in the body, and having a short half-life in the blood. This necessitates frequent administration by injection, placing a significant burden on both patients and medical professionals. Therefore, there is a demand for drug matrices (sustained-release drug delivery system matrices) that can encapsulate biopharmaceuticals without impairing their pharmacological activity and gradually release the active ingredient in the body.

[0004] For example, Patent Document 1 discloses a hyaluronic acid derivative that spontaneously associates in an aqueous solution and can efficiently encapsulate drugs, particularly biopharmaceuticals, while retaining their biological activity, as a base material for a sustained-release drug delivery system.

[0005] Patent Document 2 discloses that dietary fiber enhances the absorption efficiency of hyaluronic acid itself. When dietary fiber reaches the intestine, intestinal bacteria that produce hyaluronidase grow in the intestine, and the decomposition of hyaluronic acid is promoted, which in turn promotes the absorption of the decomposed hyaluronic acid.

[0006] Patent Document 3 discloses that a hyaluronic acid derivative modified with a cationic group and polyethylene glycol and further having a hydrophobic group introduced therein has excellent adhesion to mucous membranes and mucosal permeability.The example in this document describes that the intraocular penetration of sorafenib after instillation into rabbits was improved.However, since cationic groups are prone to nonspecific adsorption with various intracellular biomolecules, drug bases having cationic groups have a high risk of exhibiting strong cytotoxicity. [Prior art documents] [Patent documents]

[0007] [Patent Document 1] International Publication No. 2010 / 053140 [Patent Document 2] Japanese Patent Application Publication No. 2019-071878 [Patent Document 3] International Publication No. 2019 / 098393 [Non-patent literature]

[0008] [Non-Patent Document 1] Weiss et al., Macromolecules, 2023, vol.56, p.7286-7299. Summary of the Invention [Problem to be solved by the invention]

[0009] The oral absorption of drugs from drug carriers made of hyaluronic acid derivatives reported so far has not necessarily been specifically examined.

[0010] The present invention has been made in view of the above circumstances, and aims to provide a pharmaceutical composition of hyaluronic acid derivative for oral absorption that has excellent oral absorption ability of drugs. [Means for solving the problem]

[0011] That is, the present invention includes the following aspects. [1] A pharmaceutical composition of a hyaluronic acid derivative for oral absorption, comprising (A) a hyaluronic acid derivative having a steryl group introduced therein and (B) an active ingredient, wherein the content of the active ingredient (B) per 100 parts by mass of the hyaluronic acid derivative (A) is 0.1 parts by mass or more but less than 10,000 parts by mass, and the average molecular weight of the hyaluronic acid derivative (A) is 6,000 or more but 20,000 or less. [2] The pharmaceutical composition of the hyaluronic acid derivative for oral absorption according to [1], wherein the (A) hyaluronic acid derivative and the (B) active ingredient form a solid dispersion. [3] The pharmaceutical composition of an orally absorbable hyaluronic acid derivative according to [2] above, wherein the active ingredient (B) is amorphous. [4] The pharmaceutical composition of the hyaluronic acid derivative for oral absorption according to [1], wherein the (A) hyaluronic acid derivative and the (B) active ingredient form nanoparticles. [5] A pharmaceutical composition of a hyaluronic acid derivative for oral absorption according to any one of [1] to [4], wherein the content of the active ingredient (B) relative to 100 parts by mass of the hyaluronic acid derivative (A) is 10 parts by mass or more and 250 parts by mass or less. [6] The pharmaceutical composition of any one of [1] to [5] for oral absorption of a hyaluronic acid derivative, wherein the introduction rate of a steryl group in the hyaluronic acid derivative (A) is 5% or more and less than 50%. [7] The pharmaceutical composition of hyaluronic acid derivative for oral absorption according to any one of [1] to [6] above, wherein the (B) active ingredient is at least one selected from proteins, peptides, and poorly water-soluble drugs. [8] The pharmaceutical composition of any one of [1] to [7] for orally absorbable hyaluronic acid derivatives, wherein the hyaluronic acid derivative (A) has one or more repeating units represented by the following general formula (I):

[0012] [ka]

[0013] (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 -NR 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 It is a group selected from the group consisting of groups represented by -SR and -SSR. 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 R is a group selected from the group consisting of alkyl. a , R b and R c The alkyl portion of is -O- and -NR f A group selected from the group consisting of - may be inserted. R f is a hydrogen atom, C 1-12 Alkyl, Amino C 2-12 Alkyl and Hydroxy C 2-12 R is a group selected from the group consisting of alkyl. f The alkyl portion of the formula (I) may be inserted with a group selected from the group consisting of -O- and -NH-. R is a steryl group. Y is C 2-30 Alkylene, or -(CH2CH2O) m -CH2CH2-. Here, the alkylene of Y is -O-, -NR gA group selected from the group consisting of - and -SS- may be inserted. R g is a hydrogen atom, C 1-20 Alkyl, Amino C 2-20 Alkyl and Hydroxy C 2-20 R is a group selected from the group consisting of alkyl. g The alkyl portion of the formula (I) may be inserted with a group selected from the group consisting of -O- and -NH-. Y a is C 1-5 It is alkylene. Y b is C 2-8 Alkylene or C 2-8 It is alkenylene. m is an integer between 1 and 100. [9] The oral hyaluronic acid derivative pharmaceutical composition according to [8], wherein the steryl group is a cholesteryl group.

[10] A pharmaceutical composition of a hyaluronic acid derivative for oral absorption according to any one of [1] to [9], in which absorption of the active ingredient (B) is promoted in at least a part of the stomach, small intestine (duodenum, jejunum, ileum), and large intestine. [Effects of the Invention]

[0014] According to the present invention, it is possible to provide a pharmaceutical composition of a hyaluronic acid derivative for oral absorption that has excellent oral absorption ability of a drug. [Brief explanation of the drawings]

[0015] [Figure 1] 1 is a graph showing the results of XRD analysis of a solid preparation (Example 1-3) containing a hyaluronic acid derivative and cyclosporine (CyA). [Figure 2] 1 is a graph showing the results of XRD analysis of a solid preparation (Example 1-4) containing a hyaluronic acid derivative and cyclosporine (CyA). [Figure 3] 1 is a graph showing the results of XRD analysis of a solid preparation (Examples 1-5) containing a hyaluronic acid derivative and cyclosporine (CyA). [Figure 4]1 is a graph showing the time course of plasma concentration of cyclosporine (CyA). [Figure 5] 1 is a graph showing the area under the serum drug concentration-time curve (AUCall) of CyA (0 to infinity). [Figure 6] 1 is a graph showing the plasma concentration of ARV-825 over time. [Figure 7] 1 is a graph showing the area under the serum drug concentration-time curve (AUCall) of ARV-825 (0 to infinity). DETAILED DESCRIPTION OF THE INVENTION

[0016] Below, we will explain in detail the embodiment of the present invention (hereinafter referred to as the ``present embodiment''), but the present invention is not limited to this and various modifications are possible within the scope of the gist of the present invention.

[0017] The terms used in this specification will be explained below.

[0018] The term "oral absorption" as used herein means that the active ingredient (B) is absorbed in at least a part of the stomach, small intestine (duodenum, jejunum, ileum), or large intestine and transferred into the blood.

[0019] The term "oral absorption enhancement" as used herein means that (A) the hyaluronic acid derivative contributes to enhancing the oral absorption of (B) the active ingredient.

[0020] 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, iso-propyl, n-butyl, sec-butyl, iso-butyl, tert-butyl, etc. 1-4alkyl," 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, carbon number 1 to 6, C 1-6 Alkyl groups are also included.

[0021] As used herein, "C 1-6 The term "alkylcarbonyl" refers to the alkyl moiety of the already mentioned C 1-6 It means an alkylcarbonyl group, for example, acetyl, propionyl, n-propylcarbonyl, isopropylcarbonyl, n-butylcarbonyl, sec-butylcarbonyl, isobutylcarbonyl, tert-butylcarbonyl, etc. 1-4 alkylcarbonyl".

[0022] As used herein, "amino acid 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, for example, the amino group may be located on the terminal carbon atom of the alkyl group. 2-20 Alkyl includes amino C with 2 to 12 carbon atoms. 2-12 Alkyl is also included.

[0023] 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 has 2 to 12 carbon atoms and hydroxy C 2-12 Alkyl is also included.

[0024] As used herein, "C 2-30The 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 with 2 to 8 carbon atoms 2-8 Alkylene, group -(CH2) 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).

[0025] 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.

[0026] As used herein, "C 2-8 The term "alkenylene" means a linear or branched, divalent saturated hydrocarbon group containing from 2 to 8 carbon atoms and containing one or more double bonds, and includes, for example, -CH=CH-, -C(CH3)=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.

[0027] <Hyaluronic acid derivative pharmaceutical composition for oral absorption> This embodiment is a hyaluronic acid derivative pharmaceutical composition for oral absorption, which comprises (A) a hyaluronic acid derivative into which a steryl group has been introduced and (B) an active ingredient, wherein the content of the (B) active ingredient per 100 mass parts of the (A) hyaluronic acid derivative into which a steryl group has been introduced is 0.1 mass parts or more and less than 10,000 mass parts, and the average molecular weight of the (A) hyaluronic acid derivative is 6,000 to 20,000. Hereinafter, "oral absorption hyaluronic acid derivative pharmaceutical composition" may be abbreviated as "pharmaceutical composition".

[0028] (B) The active ingredient may be crystalline or amorphous. Amorphous refers to solid molecules, atoms, and / or ions that are not crystalline. Amorphous solids do not exhibit a distinct X-ray diffraction pattern. They can also be identified by the absence of endothermic peaks characteristic of poorly soluble drug crystals through thermal analysis such as differential scanning calorimetry (DSC).

[0029] In the pharmaceutical composition of this embodiment, the hyaluronic acid derivative may form a complex with the active ingredient (hereinafter, sometimes referred to as an "active ingredient-hyaluronic acid derivative complex"). Specifically, a complex may be formed through hydrophobic interaction between the steryl group in the hyaluronic acid derivative and the hydrophobic portion of the active ingredient. The form of the complex is not particularly limited and is determined appropriately based on the type of hyaluronic acid derivative and the active ingredient, specifically their chemical structure, physicochemical properties such as solubility, hydrophobicity, and ease of crystallization, and the content ratio of the hyaluronic acid derivative to the active ingredient. For example, the complex may have a cylindrical structure in which the active ingredient and hydrophobic portion, such as the steryl group, are present in the center and hydrophilic portions, such as portions derived from hyaluronic acid in the hyaluronic acid derivative, are present in the outer periphery, i.e., the active ingredient is encapsulated or encapsulated in the hyaluronic acid derivative. For example, the hyaluronic acid derivative and the active ingredient form nanoparticles.

[0030] Furthermore, the hyaluronic acid derivative and the active ingredient do not necessarily form a complex, and physical contact between the drugs may be suppressed. In other words, the hyaluronic acid derivative and the active ingredient may form a solid dispersion. In this application, "solid dispersion" refers to a solid state in which the active ingredient is dispersed in the hyaluronic acid derivative. A preferred example of the pharmaceutical composition of this embodiment is a solid dispersion composition in which an amorphous form of the active ingredient (B) is dispersed in a solid state of the hyaluronic acid derivative polymer (A). In particular, it is preferable that the amorphous active ingredient is distributed in a molecularly dispersed state within the hyaluronic acid derivative polymer (A). In this case, it is more preferable that the hyaluronic acid derivative polymers are entangled with each other, suppressing crystallization of the active ingredient, and maintaining the active ingredient in an amorphous (non-crystalline structure) state. A preparation containing a solid dispersion may have higher solubility in an aqueous medium than a crystalline active ingredient upon dissolution. Therefore, when the active ingredient (B) in the pharmaceutical composition of this embodiment is a solid dispersion, it is advantageous in that it can be easily made into a preparation with excellent absorbability in the stomach and intestinal tract.

[0031] In the pharmaceutical composition of this embodiment, when hyaluronic acid derivative and active ingredient form nanoparticles, the average particle size of the nanoparticles is not particularly limited, but can be 10nm or more, preferably 20nm or more, more preferably 30nm or more.In the pharmaceutical composition of this embodiment, the average particle size of the structure can be 10μm or less, preferably 5μm or less, more preferably 1μm or less, even more preferably 0.5μm or less, can be 250nm or less, can be 100nm or less.By having the average particle size within the above numerical range, particle dispersion stability can be expected, and the active ingredient-hyaluronic acid derivative complex can be stably present in vivo, and the effect of hyaluronic acid derivative as a drug carrier can be more strongly exerted.

[0032] In the pharmaceutical composition of this embodiment, when the hyaluronic acid derivative and the active ingredient form a solid dispersion, the average particle size of the solid dispersion when dispersed in a solvent is not particularly limited, but can be 10 nm or more, preferably 20 nm or more, and more preferably 30 nm or more. In the pharmaceutical composition of this embodiment, the average particle size of the structure can be 100 μm or less, preferably 50 μm or less, more preferably 10 μm or less, even more preferably 5 μm or less, and can be 1 μm or less, and can be 500 nm or less. When the average particle size is within the above numerical range, particle dispersion stability can be expected, and a solid dispersion consisting of the active ingredient and the hyaluronic acid derivative can be stably dispersed in vivo, thereby more effectively demonstrating the effect of the hyaluronic acid derivative as a drug carrier. In addition, the amorphous nature of the active ingredient can be confirmed, for example, by differential scanning calorimetry (DSC) or X-ray diffraction (XRD).

[0033] In the present specification, the "average particle size of nanoparticles or solid dispersions" refers to the z-average particle size measured by dynamic light scattering. The z-average particle size can be measured by dynamic light scattering using, for example, a solution in which nanoparticles or solid dispersions are dissolved or dispersed in phosphate buffer (10 mM) to a concentration of 1 mg / mL. Alternatively, particle size may be measured by laser diffraction.

[0034] Next, the components of this embodiment will be described in detail below.

[0035] <Hyaluronic acid derivatives with steryl groups> The pharmaceutical composition of this embodiment comprises (A) a hyaluronic acid derivative into which a steryl group has been introduced, and the average molecular weight of the (A) hyaluronic acid derivative is 6,000 or more and 20,000 or less. Hereinafter, "(A) a hyaluronic acid derivative into which a steryl group has been introduced" may be referred to as "(A) hyaluronic acid derivative".

[0036] The steryl group may be directly bound to hyaluronic acid or may be bound via a linker. The "linker" referred to here is a group in which a group reactive with a carboxy group and a group reactive with a steryl group are linked by a chain group (spacer). Examples of the group reactive with a carboxy group and the group reactive with a steryl group include an amino group and a hydroxyl group. Examples of the chain group include a chain hydrocarbon group, a polyethylene glycol (PEG) chain, any peptide linker that can be introduced by genetic engineering, or a synthetic compound linker. The length of the linker is not particularly limited and can be appropriately selected by those skilled in the art depending on the purpose. Examples of the chain hydrocarbon group include C 1-30 Alkylene is preferred, C 1-10 Alkylene is more preferred, C 1-6 Alkylene is more preferred. As the PEG group, a group in which the number of linked ethylene glycol groups is 1 to 15 is preferred, a group in which the number is 1 to 10 is more preferred, and a group in which the number is 1 to 5 is even more preferred. As the peptide linker, two or more amino acids (the upper limit is not particularly limited, but usually 30 or less amino acids, preferably 20 or less amino acids) are used, and 15 amino acids are particularly preferred. As the hyaluronic acid derivative of this embodiment, it is preferred that at least a portion of the carboxyl groups in the glucuronic acid moiety are linked to steryl groups via a linker having a chain-like hydrocarbon group.

[0037] (A) When the hyaluronic acid derivative has steryl groups, the steryl groups in the hyaluronic acid derivative self-associate in water, and a single molecule or multiple molecules associate to form a nano-sized hydrogel.

[0038] [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, with cholesteryl groups (particularly cholest-5-en-3β-yl groups) being preferred.

[0039] [Steryl group introduction rate] (A) The introduction rate of steryl groups into the hyaluronic acid derivative (hereinafter sometimes simply referred to as "steryl group introduction rate") is preferably 5% or more and less than 50%, more preferably 10% or more and 45% or less, even more preferably 15% or more and 45% or less, and even more preferably 25% or more and 45% or less.

[0040] When the steryl group introduction rate is within the above range, the hyaluronic acid derivative (A) can strongly interact with the hydrophobic portion of the poorly water-soluble drug. Also, when the steryl group introduction rate is within the above range, the hyaluronic acid derivative-drug complex, which is the complex of the hyaluronic acid derivative (A) and the drug in the pharmaceutical composition, improves the stability of the formulation and aggregates and precipitates under physiological salt concentrations, enabling sustained release of the drug.

[0041] The steryl group introduction rate is 1 It can be measured by H-NMR measurement. 1 The H-NMR spectrum can be calculated based on the following formula using the integral value of the peak derived from the steryl group of (A) the hyaluronic acid derivative and the integral value of the peak derived from the acetyl group of N-acetyl-D-glucosamine contained in (A) the hyaluronic acid derivative (COCH3, 1.6 ppm to 2.0 ppm, 3H).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.

[0042] [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

[0043] The average molecular weight of (A) hyaluronic acid derivative is sufficient as long as it is 6,000 (6k) or more and 20,000 (20k) or less, and can be adjusted appropriately according to the purpose and dosage form.For example, from the viewpoint of improving the sustained release function derived from the diffusion delay in oral administration, (A) hyaluronic acid derivative with a relatively large molecular weight is preferred.On the other hand, when the final dosage form is a solution preparation, from the viewpoint of syringeability, (A) hyaluronic acid derivative with a relatively small molecular weight is preferred.

[0044] (A) The average molecular weight of hyaluronic acid derivative is preferably 7k or more and 18k or less, more preferably 8k or more and 16k or less.By making the weight-average molecular weight of hyaluronic acid derivative be above-mentioned lower limit value or more, it can suppress the increase of viscosity, and can dissolve the hyaluronic acid derivative in pharmaceutical composition at higher concentration.

[0045] The "average 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). The weight-average molecular weight of the hyaluronic acid derivative can generally be adjusted by using a raw material having a corresponding molecular weight.

[0046] Specific examples of preferred (A) hyaluronic acid derivatives include hyaluronic acid derivatives having one or more repeating units represented by the following general formula (I) (hereinafter, sometimes referred to as "repeating unit (I)").

[0047] [ka]

[0048] (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 -NR 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 It is a group selected from the group consisting of groups represented by -SR and -SSR. 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 R a , R b and R c The alkyl portion of is -O- and -NR f A group selected from the group consisting of - may be inserted. R f is a hydrogen atom, C 1-12 Alkyl, Amino C 2-12 Alkyl and Hydroxy C 2-12 R is a group selected from the group consisting of alkyl. f The alkyl portion of the formula (I) may be inserted with a group selected from the group consisting of -O- and -NH-. R is a steryl group. Y is C 2-30 Alkylene, or -(CH2CH2O) m -CH2CH2-. Here, the alkylene of Y is -O-, -NR g A group selected from the group consisting of - and -SS- may be inserted. R g is a hydrogen atom, C 1-20 Alkyl, Amino C 2-20 Alkyl and Hydroxy C 2-20 R is a group selected from the group consisting of alkyl. g The alkyl portion of the formula (I) may be inserted with a group selected from the group consisting of -O- and -NH-. Y a is C 1-5 It is alkylene. Y b is C 2-8 Alkylene or C 2-8 It is alkenylene. m is an integer between 1 and 100.

[0049] The hyaluronic acid derivative (A) 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)").

[0050] [ka]

[0051] (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 X is a group selected from the group consisting of -NR a -Y-NR b -COO-R is a hydrophobic group. a and R b are each independently a hydrogen atom and C 1-6alkyl; R is a steryl group; Y is C 2-30 Alkylene, or -(CH2CH2O) m -CH2CH2-, and m is an integer of 1 or more and 100 or less.

[0052] 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.

[0053] 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 to an amide or ester, or may form a salt.

[0054] Repeating Unit The group "-ZN(R a )YX 1 " is the formula: -NH-(CH2) mz -NH-R; -NH-(CH2) mz -NH-COO-R; -NH-(CH2CH2O) m -CH2CH2-NH-COO-R; -NH-(CH2) mz -COO-R; -NH-(CH2CH2O) m -CH2CH2-COO-R, -NH-(CH2) mz -O-COO-R; -NH-(CH2CH2O) m -CH2CH2-O-COO-R, -NH-(CH2) mz -SR; -NH-(CH2CH2O) m -CH2CH2-SR; -NH-(CH2) mz -O-CO-CH(R 8 )-CH2-SR; -NH-(CH2) mz -NHCO-CH(R 8 )-CH2-SR; -NH-(CH2CH2O) m -CH2CH2-NHCO-CH(R 8 )-CH2-SR; -NH-(CH2CH2O) m -CH2CH2-O-CO-CH(R 8 )-CH2-SR; -NH-(CH2) mz -SSR; and -Z-NR a -Y-NR b -COO-R (where mz is an integer between 2 and 30, and R 8 is a hydrogen atom or a methyl group, and R and m are as defined above in this specification. The group includes a group selected from the group consisting of groups represented by the formula: Examples of the group include: -NH-(CH2) mz -NH-COO-R; -NH-(CH2CH2O) m -CH2CH2-NH-COO-R; and -NH-(CH2) mz -SSR (wherein mz, ​​R, and m are as defined previously in this specification.) A group selected from the group consisting of: is preferred.

[0055] (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 Preferably, Z is -COO-R. In another embodiment, Z is -NH-[CH(-Z a )-CONH] n-1 -CH(-Z a)-CO-, where n is an integer of 2 or more and 30 or less, and Z a are each independently H2N-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 )-YX 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. That is, Z a Examples of such linkers include -CH3, H2NC(NH)NH(CH2)3-, H2NCOCH2-, and the like. Furthermore, n Zs may be the same or different. n is an integer of 2 to 30, preferably 2 to 10, and more preferably 2 to 4. Preferred examples of peptide linkers include -Gly-Phe-Leu-Gly-, -Asn-Phe-Phe-, -Phe-Phe-, Phe-Gly-, and the like.

[0056] (Y) In the general formula (I), Y is —(CH) n1 -and-(CH2CH2O) m1 A group selected from the group consisting of -CH2CH2- (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, -(CH2)2-, -(CH2)6-, -(CH2)8-, -(CH2) 12-, or -(CH2CH2O)2-CH2CH2- is preferred. From the viewpoint of realizing high solubility in pure water or at low salt concentrations and exhibiting high precipitate-forming ability at physiological salt concentrations, Y is preferably -(CH2)2-, -(CH2)6-, -(CH2)8-, or -(CH2) 12 A group selected from the group consisting of - is preferred, with -(CH2)6- being more preferred.

[0057] Y may be, for example, -CH2CH2O-CH2CH2-SS-CH2CH2O-CH2CH2-, -(CH2CH2O)2-CH2CH2-SS-CH2CH2O-CH2CH2-, -CH2CH2O-CH2CH2-SS-(CH2CH2O)2-CH2CH2-, -(CH2CH2O)2-CH2CH2-SS-(CH2CH2O)2-CH2CH2-, -(CH2CH2O)2-CH2CH2-SS-(CH2CH2O)2-CH2CH2-, and the like.

[0058] (Y a ) Y a is preferably -CH2- or -CH2-CH2-.

[0059] (Y b ) Y b As the alkyl group, -CH2-CH2-, -CH(CH3)CH2-, 2-butene-1,4-diyl, hepta-2,4-diene-1,6-diyl, or octa-2,4,6-triene-1,8-diyl is preferred, and -CH2-CH2- or -CH(CH3)CH2- is more preferred.

[0060] Group "-ZN(R a )YX 1" is exemplified by -NH-(CH2)2-NH-CO-cholesteryl, -NH-(CH2)4-NH-(CH2)3-NH-(CH2)3-NH-COO-cholesteryl, -NH-(CH2)3-NH-(CH2)4-NH-(CH2)3-NH-COO-cholesteryl, -NH-(CH2)4-NH-(CH2)3-NH-COO-cholesteryl, -NH-(CH2)4-NH-(CH2)3-NH-COO-cholesteryl, -NH-(CH2)4-N(-(CH2)3-NH2)-COO-cholesteryl, -NH-(CH2)3-NH-(CH2)4-N(-(C -NH-(CH)-NH-(CH)-COO-cholesteryl, -NH-(CH)-NH-(CH)-N(-(CH)-NH-(CH)-NH)-COO-cholesteryl, -NH-(CH)-NH-(CH)-N(-(CH)-NH)-CO-NH-cholesteryl, -NH-(CH)-NH-(CH)-N(-(CH)-NH)-CO-cholesteryl, -NH-(CH)-NH-(CH)-N(-(CH)-NH)-cholesteryl, and the like are preferred. a )YX 1 " as R a , R b and R c is a hydrogen atom, and Y is a linear C 2-30 Alkylene or -(CH2CH2O) m -CH2CH2-, and Y a However, linear C 1-5 alkylene, or Y b However, linear C 2-8 Alkylene or linear C 2-8 It is alkenylene.

[0061] Repeating unit (Ia) In the general formula (Ia), X is -NH-(CH2)2-NH-COO-cholesteryl, -NH-(CH2)6-NH-COO-cholesteryl, -NH-(CH2) 12-NH-COO-cholesteryl or -NH-(CH2CH2O)2-CH2CH2-NH-COO-cholesteryl is preferred, and -NH-(CH2)2-NH-COO-cholesteryl, -NH-(CH2)6-NH-COO-cholesteryl or -NH-(CH2CH2O)2-CH2CH2-NH-COO-cholesteryl is more preferred.

[0062] [Repeating units incorporating maleimide groups] (A) The hyaluronic acid derivative may contain, in addition to the repeating unit (I), a structural unit into which a maleimide group (N-maleimide group) has been introduced. The maleimide group may be directly bonded to the hyaluronic acid, or may be bonded via a linker. The linkers connected to the maleimide groups contained in one molecule of the hyaluronic acid derivative may all be the same linker, or may be linkers of different lengths or types.

[0063] The "linker" referred to here is a group in which a group reactive with a carboxy group and a group reactive with an NH group in a maleimide group are linked by a chain group. Examples of the group reactive with a carboxy group include an amino group and a hydroxyl group, and examples of the group reactive with an NH group in a maleimide group include a carboxy group and a hydroxyl group. Examples of the chain group include the same group as the chain group in the linker that links the hyaluronic acid derivative and the steryl group. In the hyaluronic acid derivative of this embodiment, it is preferred that at least a part of the carboxyl group in the glucuronic acid moiety and the hydroxyl group in the N-acetylglucosamine moiety are linked to the NH group in the maleimide group by a linker having a chain hydrocarbon group.

[0064] An example of a repeating unit into which a maleimide group has been introduced is a repeating unit represented by general formula (II) (hereinafter, sometimes referred to as "repeating unit (II)").

[0065] [ka]

[0066] (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 2 is a maleimide group. R d is a hydrogen atom, C 1-20 Alkyl, Amino C 2-20 Alkyl and Hydroxy C 2-20 alkyl, and R d The alkyl portion of is -O- and -NR f A group selected from the group consisting of - may be inserted. R f is a hydrogen atom, C 1-12 Alkyl, Amino C 2-12 Alkyl and Hydroxy C 2-12 R is a group selected from the group consisting of alkyl. f The alkyl portion of the formula (I) may be inserted with a group selected from the group consisting of -O- and -NH-. Y is C 2-30 Alkylene, or -(CH2CH2O) m -CH2CH2-. Here, the alkylene of Y is -O-, -NR g A group selected from the group consisting of - and -SS- may be inserted. R g is a hydrogen atom, C 1-20 Alkyl, Amino C 2-20 Alkyl and Hydroxy C 2-20 R is a group selected from the group consisting of alkyl. g The alkyl portion of the formula (I) may be inserted with a group selected from the group consisting of -O- and -NH-. m is an integer between 1 and 100.

[0067] Here, when the hyaluronic acid derivative (A) contains two or more repeating units (II), the repeating units may be the same or different. In another embodiment, the hyaluronic acid derivative (A) may be a hyaluronic acid derivative consisting essentially of the repeating unit (I), the repeating unit (Ia) and the repeating unit (II).

[0068] The repeating unit (II) is particularly a repeating unit represented by the general formula (II), in which Z is a direct bond and R d is a hydrogen atom or C 1-6 alkyl and Y is C 2-30 Alkylene or -(CH2CH2O) m -CH2CH2- and X 2 is a maleimide group; Z is a direct bond, and R d is a hydrogen atom or C 1-6 alkyl and Y is C 2-30 is alkylene, and X 2 is a maleimide group; Z is a direct bond and R d is a hydrogen atom and Y is C 2-30 is alkylene, and X 2 is a maleimide group; Z is a direct bond and R d is a hydrogen atom and Y is C 2-10 is alkylene, and X 2 It is more preferable that the repeating unit is a maleimide group.

[0069] In the pharmaceutical composition of this embodiment, when the maleimide group is introduced into the hyaluronic acid derivative (A) in the present embodiment, the introduction rate of the maleimide group in the hyaluronic acid derivative (hereinafter, sometimes simply referred to as "maleimide group introduction rate") is not particularly limited, but is preferably 1.0% or more, more preferably 3.0% or more, more preferably 5.0% or more, and even more preferably 10% or more. The introduction rate of the maleimide group in the hyaluronic acid derivative (A) is preferably 25% or less, more preferably 20% or less.

[0070] The maleimide group introduction rate is 1 It can be measured by H-NMR measurement. 1 The H-NMR spectrum is obtained by measuring a solution in which the hyaluronic acid derivative of this embodiment is dissolved in a heavy solvent 0.02N DCl DMSO-d6 / D2O mixed solution (2N DCl D2O:DMSO-d6=1:99). 1 The introduction rate of cholesteryl groups and maleimide groups to the hyaluronic acid unit is calculated using the integral value of the peak (COCH3, 1.6-2.0 ppm; 3H) derived from the acetyl group of the glucosamine portion of the hyaluronic acid derivative in the H-NMR spectrum, the integral value of the peak (CH3, 0.7 ppm; 3H) derived from the methyl group in the cholesteryl group, and the integral value of the peak (-CH=CH-, 6.9 ppm; 2H) derived from the maleimide group, according to the following formula: Note that the peak (5H) derived from cholesteryl groups overlaps with the peak near 1.6-2.0 ppm, which includes the peak derived from the acetyl group of the glucosamine moiety. Therefore, the value calculated by subtracting 5 / 3 times the integral value of the peak derived from the cholesteryl group methyl (0.7 ppm) from the integral value of the peak near 1.6-2.0 ppm (i.e., [[peak integral value (1.6-2.0 ppm)] - [peak integral value (0.7 ppm)] × 5 / 3]) is used as the integral value (corrected value) of the acetyl group derived from hyaluronic acid to calculate the introduction rate. First, the cholesteryl group introduction rate (%) is calculated using the above-mentioned formula for calculating the steryl group introduction rate.

[0071] Furthermore, the maleimide group introduction rate (%) can be calculated based on the following formula using the integral value of the peak derived from the maleimide group and the integral value of the peak derived from the cholesteryl group (COCH, 1.6 ppm to 2.0 ppm, 3H): Specifically, it can be measured according to the method described in the Examples below.

[0072] [Maleimide group introduction rate (%)] = ([Integrated value of peaks derived from maleimide groups] × 3) / ([Integrated value of peaks derived from cholesteryl groups] × 2) × [Cholesteryl group introduction rate (%)]

[0073] The content of the hyaluronic acid derivative (A) relative to the total amount of the pharmaceutical composition of this embodiment is preferably 1 mg / mL or more and less than 50 mg / mL, more preferably 3 mg / mL or more and 45 mg / mL or less, and even more preferably 5 mg / mL or more and 40 mg / mL or less.

[0074] (Method of producing hyaluronic acid derivatives) The hyaluronic acid derivative (A) contained in the pharmaceutical composition of this embodiment can be obtained, for example, by converting the carboxy group of glucuronic acid to an amide and introducing a steryl group directly or via a linker into at least a portion of the hyaluronic acid derivative. Alternatively, the hyaluronic acid derivative can be obtained by converting the carboxy group of glucuronic acid to an amide, introducing a steryl group directly or via a linker into at least a portion of the hyaluronic acid derivative, and introducing a maleimide group directly or via a linker into the remaining portion of the hyaluronic acid derivative. The steryl group introduction rate can be controlled by adjusting the amount of the compound having a steryl group to be reacted with the raw material hyaluronic acid or its derivative. The maleimide group introduction rate can be controlled by adjusting the amount of the compound having a maleimide group to be reacted. Methods for introducing maleimide groups or steryl groups into the raw material hyaluronic acid can be carried out by appropriately modifying the methods described in, for example, JP 2021-123597 A, JP 2022-013861 A, JP 2022-044579 A, ​​etc.

[0075] (A) When both a steryl group and a maleimide group are introduced into the hyaluronic acid derivative, the raw material hyaluronic acid may be reacted with a compound having a steryl group, and the resulting reaction product may be reacted with a compound having a maleimide group; the raw material hyaluronic acid may be reacted with a compound having a maleimide group, and the resulting reaction product may be reacted with a compound having a steryl group; or the raw material hyaluronic acid may be added to the reaction system together with a compound having a maleimide group and a compound having a steryl group, and reacted with the resulting reaction product.

[0076] A specific example of a method for converting the carboxy group of glucuronic acid to an amide and introducing a steryl group or maleimide group is to ion-exchange the starting hyaluronic acid or a derivative thereof with a tetraalkylammonium salt (e.g., tetrabutylammonium (TBA) salt), and then react the hyaluronic acid salt with an amine having a steryl group (particularly a cholesteryl group) in a solvent in the presence of a suitable condensing agent.

[0077] 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).

[0078] In particular, although not limited to, DMT-MM is preferred because the reaction proceeds efficiently even in a mixed solvent of water and an organic solvent.In addition, by using DMT-MM as a condensing agent, in a system where a large number of hydroxyl groups coexist, it is possible to suppress the formation of ester bonds and highly selectively form amide bonds between amino groups and carboxyl groups.The use of this condensing agent can, for example, prevent the reaction of the solvent alcohol with the carboxyl group of the hyaluronic acid moiety, or prevent the intramolecular or intermolecular bonding of the carboxyl group and the hydroxyl group simultaneously present in the hyaluronic acid moiety, resulting in the formation of undesired crosslinks.

[0079] 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 mixtures 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.

[0080] In the production of hyaluronic acid derivatives into which both steryl and maleimide groups have been introduced, the pH of the reaction system is preferably acidic during the steryl group introduction reaction, whereas under basic conditions, succinimide groups may be generated by the deactivation of the maleimide groups.

[0081] Alternatively, the starting hyaluronic acid or its derivative can be ion-exchanged with a tetraalkylammonium salt (e.g., tetrabutylammonium (TBA) salt), and then react this hyaluronic acid salt with a spacer moiety in a solvent in the presence of a suitable condensing agent (at this time, protection and deprotection reactions can be carried out as necessary), converting the carboxyl group (-COOH) of the starting hyaluronic acid or its derivative, and then reacting it with a suitable reagent. Examples of the combination of groups derived from carboxyl 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 bH + 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-NR b 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=CH2+ HS-R; -CONR a -Y-NR b -CO-CH(CH3)=CH2+ 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 (In the formula, 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).

[0082] 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 reaction product has a carboxy group, it may be converted into an N-hydroxysuccinimide (hereinafter also referred to as "NHS") ester and then reacted.

[0083] Another method is to react the carboxyl group of the raw material hyaluronic acid or its derivative with 2-aminoethyl 2-pyridyl disulfide to prepare a hyaluronic acid derivative having a spacer with a mercapto group modified with a leaving group at the end, and then subject this to a nucleophilic substitution reaction with thiocholesterol to form a disulfide bond.

[0084] Furthermore, the method of preparing the carboxyl group of hyaluronic acid or its derivatives that introduces a part of spacer, and the method of introducing a part of spacer into steryl group, and then reacting them.Some specific examples have been mentioned above, but when -SS- is inserted into Y, the carboxyl group of hyaluronic acid can also be prepared by introducing the spacer that has mercapto group at the end of the hyaluronic acid derivative, and the steryl group that has mercapto group at the end of the spacer, and then oxidatively react them to form disulfide bond.At this time, one mercapto group can be reacted with 2-mercaptopyridine to form disulfide, and then be replaced with the other mercapto group.

[0085] After preparing the hyaluronic acid derivative, other substituents may be introduced. For example, 0.1% to 95.0%, preferably 10% to 60%, of the carboxy groups in the hyaluronic acid derivative may be replaced with -CO-X z , where X z is the following group: -NH-(CH2) p1 -O-CO-C(R 17 )=CH2; -NH-(CH2) p1 -O-CO-CH(R 17 )-CH2-S-CH2-CH(OH)-CH(OH)-CH2-SH; -NH-(CH2) p1 -SH; -NH-(CH2) p1 -NH-CO-C(R 17 )=CH2; -NH-(CH2) p1 -NH-C(=NH)-(CH2)3-SH; -NH-(CH2) p1 -NH-CO-(CH2) r -SH; -NH-(CH2) p1 -NH-CO-CH(R 17 )-CH2-S-CH2-CH(OH)-CH(OH)-CH2-SH; -NH-(CH2) p1 -NH-CO-CH(NH2)-CH2-SH; -NH-(CH2) p1 -NH-CO-CH(NH2)-(CH2)2-SH; -NH-NH-CO-(CH2)4-CO-NH-NH-C(=NH)-(CH2)3-SH; -NH-(CH2-CH2-O) q -CH2-CH2-O-CO-C(R 17 )=CH2; -NH-(CH2-CH2-O) q -CH2-CH2-O-CO-CH(R 17 )-CH2-S-CH2-CH(OH)-CH(OH)-CH2-SH; -NH-(CH2-CH2-O) q -CH2-CH2-SH; -NH-(CH2-CH2-O) q -CH2-CH2-NH-CO-C(R 17 )=CH2; -NH-(CH2-CH2-O) q -CH2-CH2-NH-C(=NH)-(CH2)3-SH; -NH-(CH2-CH2-O) q -CH2-CH2-NH-CO-(CH2) r -SH; -NH-(CH2-CH2-O) q -CH2-CH2-NH-CO-CH(R 17 )-CH2-S-CH2-CH(OH)-CH(OH)-CH2-SH; -NH-(CH2-CH2-O) q -CH2-CH2-NH-CO-CH(NH2)-CH2-SH; -NH-(CH2-CH2-O) q -CH2-CH2-NH-CO-CH(NH2)-(CH2)2-SH; -NH-CH(CO2H)-(CH2)-SH; -NH-CH(COH)-(CH)-SH; and -NH-CH(CO2H)-(CH2)2-CONH-CH(CONH-CH2-CO2H)-CH2-SH (where R 17 is a hydrogen atom or C 1-6 p1 is an alkyl group, p2 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. It can also be converted to

[0086] (B) Active ingredient The pharmaceutical composition of this embodiment comprises (A) a hyaluronic acid derivative and (B) an active ingredient.The active ingredient (B) in the pharmaceutical composition of this embodiment is not particularly limited, as long as it is a drug for humans or animals (a substance administered for the diagnosis, treatment, or prevention of disease) or a substance used as an active ingredient thereof.

[0087] The diseases for which the pharmaceutical composition of this embodiment is applicable are not particularly limited, and can be widely used for the prevention and treatment of currently known diseases or diseases that will be discovered in the future. These diseases may be chronic or acute. Examples of currently known diseases include cancer, infectious diseases, immune diseases, inflammatory diseases, allergic diseases, skin diseases, hypertension, diabetes, neurological diseases, genetic diseases, cardiovascular diseases, cerebrovascular diseases, respiratory diseases, eye diseases, ear diseases, and bone and joint diseases.

[0088] (B) The active ingredient is not particularly limited, and various substances such as pharmaceutically active peptides or proteins, nucleic acids, low molecular weight compounds, and medium molecular weight compounds can be used. A low molecular weight compound refers to a compound other than nucleic acids, peptides, and proteins, which has a molecular weight of less than about 500. A medium molecular weight compound refers to a compound other than nucleic acids, peptides, and proteins, which is neither a low molecular weight compound (organic compounds with a molecular weight of up to about 500) nor a high molecular weight compound (such as proteins with a molecular weight of 10,000 or more), but has a molecular weight of about 500 to 5,000.

[0089] [Pharmaceutically active peptides or proteins] A pharmaceutically active peptide or protein refers to one that, when administered to a subject in a therapeutically effective amount, has a positive or beneficial effect on the subject's condition or pathology. Preferred pharmaceutically active peptides or proteins have curative or palliative properties and can be administered to ameliorate, alleviate, relieve, reverse, delay the onset of, or reduce the severity of one or more symptoms of a disease or disorder. A pharmaceutically active peptide or protein may also have preventative properties and can be used to delay the onset of a disease or reduce the severity of such a disease or pathology. The term "pharmaceutically active peptide or protein" encompasses a full-length protein or polypeptide and may also refer to a pharmaceutically active fragment thereof. The term also encompasses pharmaceutically active analogs of the peptide or protein.

[0090] In one embodiment, the pharmaceutically active peptide has, for example, 8 to 120 amino acids, preferably 8 to 80 amino acids, more preferably 15 to 80 amino acids, even more preferably 16 to 80 amino acids, even more preferably 23 to 80 amino acids, still more preferably 23 to 60 amino acids, and particularly preferably 23 to 50 amino acids.

[0091] In one embodiment, the molecular weight of the pharmaceutically active peptide is preferably about 500 to 5000. A peptide with a molecular weight of about 500 to 5000 is a linear or cyclic peptide with about 5 to 50 amino acid residues. Cyclic peptides are preferred as peptides.

[0092] Examples of pharmaceutically active proteins include, but are not limited to, cytokines and immune system proteins such as immunoactive compounds (e.g., interleukins, colony-stimulating factors (CSF), granulocyte colony-stimulating factors (G-CSF), granulocyte-macrophage colony-stimulating factors (GM-CSF), erythropoietin, tumor necrosis factors (TNF), interferons, integrins, addressins, seletins, homing receptors, T cell receptors, immunoglobulins, antibodies, hormones (insulin, thyroid hormones, catecholamines, gonadotropins, stimulating hormones, prolactin, oxytocin, dopamine, bovine somatotropin, leptin, etc.), growth hormones (e.g., human growth hormone), growth factors (e.g., epidermal growth factor, nerve growth factor, insulin-like growth factor, etc.), growth factor receptors, enzymes (tissue plasminogen activator, streptokinase, cholesterol biosynthetic enzymes or degradative enzymes, steroidogenic enzymes, kinases, phosphodiesterases, methylases, demethylases, dehydrogenases, cellulases, proteases, lipases, phospholipases, aromatase, cytochromes, adenylate cyclase or guanylate cyclase, neuramidase, etc.), receptors (steroid hormone receptors, peptide receptors), binding proteins (growth hormone binding proteins or growth factor binding proteins, etc.), transcription factors and translation factors, tumor growth suppressor proteins (for example, proteins that inhibit angiogenesis), structural proteins (collagen, fibroin, fibrinogen, elastin, tubulin, actin, myosin, etc.), blood proteins (thrombin, serum albumin, factor VII, factor VIII, insulin, factor IX, factor X, tissue plasminogen activator, protein C, von Willebrand factor, antithrombin III, glucocerebrosidase, erythropoietin, modified factor VIII, anticoagulant factors), etc.

[0093] Examples of pharmaceutically active proteins include antigens (cancer antigens, antigens derived from infectious diseases, autoantigens in immune diseases, etc.). Cancer antigens are antigens that are highly expressed on, and in some cases exclusively by, cancer cells. Cancer antigens can be expressed within or on the surface of cancer cells. The infectious disease-derived antigen is not particularly limited as long as it is an infectious pathogen and an antigen derived from an infectious pathogen.The infectious pathogen includes viruses, bacteria, fungi, nematodes, etc.The infectious disease pathogen-derived antigen is not particularly limited as long as it is at least a part of the various components constituting the pathogen, and examples thereof include live vaccines, inactivated whole particles, parts thereof, protein subunits, proteins, peptides, etc.Among them, protein subunits, proteins, or peptides are preferred from the viewpoint of complexation with hyaluronic acid derivatives. Antigens for immune diseases are not particularly limited as long as they contain an epitope of a target protein of the immune disease. Examples of immune diseases include, but are not limited to, psoriasis vulgaris, ankylosing spondylitis, rheumatoid arthritis, psoriatic arthritis, axial spondyloarthritis, Crohn's disease, ulcerative colitis, bronchial asthma, chronic urticaria, hay fever, atopic dermatitis, etc.

[0094] [Nucleic acid] Nucleic acids include DNA and RNA, such as short interfering RNA (siRNA), double-stranded RNA (dsRNA), microRNA (miRNA), short hairpin RNA (shRNA), and nucleic acid aptamers.

[0095] [Low and medium molecular weight compounds] Examples of low molecular weight compounds or medium molecular weight compounds include anticancer agents (e.g., alkylating agents, antimetabolites, alkaloids, etc.), immunosuppressants, anti-inflammatory agents (steroids, non-steroidal anti-inflammatory agents, etc.), antirheumatic agents, antibacterial agents (β-lactam antibiotics, aminoglycoside antibiotics, macrolide antibiotics, tetracycline antibiotics, new quinolone antibiotics, sulfa drugs, etc.).

[0096] (B) As active ingredient, because it can fully exert the interaction with the steryl group of the above-mentioned hyaluronic acid derivative, it is preferred to have high hydrophobicity, that is, poorly water-soluble.In addition, " poorly water-soluble" refers to the drug that is classified as very soluble, easily soluble, slightly soluble, slightly poorly soluble, poorly soluble, extremely poorly soluble or almost insoluble in the solubility terms that are listed in the 17th revised Japanese Pharmacopoeia, among which very soluble, easily soluble, slightly soluble, poorly soluble, poorly soluble, extremely poorly soluble or almost insoluble.

[0097] Specifically, the active ingredient (B) may be a poorly water-soluble drug having a solubility in water of 1 mg / mL or less. The pharmaceutical composition of the present embodiment can solubilize such poorly water-soluble drugs at high concentrations without using organic solvents and while reducing the amount of highly toxic surfactants used.

[0098] The molecular weight of the (B) active ingredient is preferably 200 or more, more preferably 300 or more, and even more preferably 400 or more, 500 or more, 600 or more, 700 or more, 800 or more, or 900 or more. The molecular weight of the (B) active ingredient may be 1000 or more, 1100 or more, or 1200 or more. The molecular weight of the (B) active ingredient is preferably 10,000 or less, more preferably 8,000 or less, and even more preferably 7,000 or less, 6,000 or less, or 5,000 or less.

[0099] (B) The molecular weight of the active ingredient is a value calculated from the molecular formula of the compound.

[0100] Examples of poorly water-soluble solid active ingredients include antipyretics and analgesics such as acetaminophen, ibuprofen, benzoic acid, ethenzamide, caffeine, camphor, quinine, calcium gluconate, dimethylcaprol, sulfamine, theophylline, theopromine, riboflavin, mephenesin, phenobarbital, aminophylline, thioacetazone, quercetin, rutin, salicylic acid, theophylline sodium salt, pyrapital, quinine hydrochloride, irgapyrin, digitoxin, griseofulvin, and phenacetin, nervous system drugs, sedatives and hypnotics, muscle relaxants, blood pressure sclerosing agents, and antihistamines; acetylspiramycin, ampicillin, erythromycin, xatamycin, chloramphenicol, triacetate, etc. Active pharmaceutical ingredients listed in the Japanese Pharmacopoeia, USP, NF, and EP include antibiotics such as tyluoleandomycin, nystatin, and colistin sulfate; steroid hormones such as methyltestosterone, methylandrostrone diol, progesterone, estradiol benzoate, ethinylestradiol, deoxycorticosterone acetate, cortisone acetate, hydrocortisone, and prednisolone; nonsteroidal yolk hormones such as dienstrol, hexastrol, diethylstilbesterol, diethylstilbesterol dibromohydratate, and chlorotrianisene; and other fat-soluble vitamins. These active ingredients may be used alone or in combination.

[0101] (B) The active ingredient may be in a water-insoluble oil or liquid form. Examples of water-insoluble oil or liquid active ingredients include teprenone, indomethacin farnesyl, menatetrenone, phytonadione, vitamin A oil, phenipentol, vitamin D, vitamin E, and other vitamins; higher unsaturated fatty acids such as DHA (docosahexaenoic acid), EPA (eicosapentaenoic acid), and liver oil; coenzyme Q; and oil-soluble flavorings such as orange oil, lemon oil, and peppermint oil, all of which are listed in the Japanese Pharmacopoeia, USP, NF, and EP. Vitamin E includes various homologs and derivatives, and is not particularly limited as long as it is liquid at room temperature. Examples include dl-α-tocopherol, dl-α-tocopherol acetate, d-α-tocopherol, and d-α-tocopherol acetate. One or more of these active ingredients may be used alone or in combination.

[0102] (B) The active ingredient may be a poorly water-soluble semi-solid active ingredient. Examples of poorly water-soluble semi-solid active ingredients include Chinese herbal or crude drug extracts such as earth dragon root, licorice, cinnamon bark, peony root, moutan pea, valerian, Japanese pepper, ginger, tangerine peel, ephedra, nandina fruit, scutellaria, onion root, platycodon, rhododendron, rhododendron bark, rhododendron japonica, garlic, seneca, fritillary root, fennel, Phellodendron bark, coptis, zedoary, chamomile, gentian, bezoar, animal gall, siberian rhizome, ginger, atractylodes rhizome, clove, tangerine peel, atractylodes rhizome, chikusetsuninjin, ginseng, pudding, keishito, kososan, shihukeishito, xiaozihuto, shoseiryuto, bakumondoto, hangehouboto, and maoto; oyster meat extract, propolis and propolis extract, and coenzyme Q. One of these active ingredients may be used, or two or more of them may be used in combination.

[0103] (B) The poorly water-soluble drug that is the active ingredient is preferably a poorly water-soluble peptide or a compound having a site that targets E3 ligase.

[0104] (B) The poorly water-soluble peptide serving as an active ingredient may contain acidic, basic, or neutral amino acids, with basic or neutral amino acids being preferred. During complexation, the pH of the solution may be appropriately selected taking into consideration the isoelectric point. Furthermore, the amino acids constituting the poorly water-soluble peptide may consist solely of natural amino acids, or unnatural amino acids, or may contain both.

[0105] (B) It is preferable that at least one of the nitrogen atoms constituting the amide bond of the poorly water-soluble peptide serving as the active ingredient has a methyl group. The hydrophobicity imparted by methylation is expected to enhance the interaction with the hydrophobic portion of the hyaluronic acid derivative, resulting in greater solubilization. It is also expected that the enhanced interaction will improve the stability of the formulation.

[0106] (B) The water-insoluble peptide serving as an active ingredient preferably comprises at least one selected from cyclic peptides and long-chain peptides, and more preferably is a cyclic peptide.The cyclic and rigid backbone maximizes the interaction with the hydrophobic part in the hyaluronic acid derivative, and forms a stable complex structure with the hyaluronic acid derivative.The cyclic size is preferably a cyclic peptide consisting of 4 to 49 amino acids, more preferably 6 to 30, and most preferably 8 to 25.

[0107] The compound having an E3 ligase targeting moiety is not particularly limited as long as it has the moiety. For example, it may be a compound consisting of only an E3 ligase targeting moiety, or a compound in which the E3 ligase targeting moiety has been modified in various ways. For example, a compound in which an E3 ligase targeting moiety and a moiety that binds to a target protein to be degraded by E3 ligase are linked directly or indirectly via a linker is preferred. Such compounds include proteolysis-targeting chimeric protein compounds (PROTACs).

[0108] Compounds having a site that targets E3 ligase include dBET1 (CAS number: 1799711-21-9), dBET6 (CAS number: 1950634-92-0), CC-885 (CAS number: 1010100-07-8), ARV-110 (CAS number: 2222112-77-6), ARV-825 (CAS number: 1818885-28-7), DT2216 (CAS number: 2365172-42-3), CC220 (CAS number: 1323403-33-3), ARCC-4 (CAS number: 1973403-00-7), and MZ-1 (CAS number: 1797406-69-9).

[0109] In this embodiment, the active ingredient (B) is not particularly limited, but may be, for example, a compound of Class 1, Class 2, Class 3, or Class 4 of the Biopharmaceutical Classification System (BCS), more preferably a compound of Class 2 (low solubility) or Class 4 (low solubility and low bioavailability).

[0110] In this embodiment, the content of the (B) active ingredient per 100 parts by mass of the (A) hyaluronic acid derivative is 0.1 parts by mass or more but less than 10,0000 parts by mass, preferably 10 parts by mass or more but less than 10,000 parts by mass, preferably 10 parts by mass or more but less than 5,000 parts by mass, more preferably 10 parts by mass or more but less than 1,000 parts by mass, even more preferably 10 parts by mass or more but less than 500 parts by mass, even more preferably 10 parts by mass or more but less than 250 parts by mass, and particularly preferably 10 parts by mass or more but less than 100 parts by mass.

[0111] In this embodiment, when the (A) hyaluronic acid derivative and the (B) active ingredient form nanoparticles, the content of the (B) active ingredient per 100 parts by mass of the (A) hyaluronic acid derivative is 0.1 parts by mass or more but less than 10,000 parts by mass, preferably 0.1 parts by mass or more but less than 30 parts by mass, more preferably 5 parts by mass or more but less than 25 parts by mass, and even more preferably 10 parts by mass or more but less than 20 parts by mass.

[0112] In this embodiment, when the (A) hyaluronic acid derivative and the (B) active ingredient form a solid dispersion, the content of the (B) active ingredient per 100 parts by mass of the (A) hyaluronic acid derivative is 0.1 parts by mass or more but less than 10,0000 parts by mass, preferably 10 parts by mass or more but less than 100,000 parts by mass, more preferably 10 parts by mass or more but less than 1,000 parts by mass, more preferably 20 parts by mass or more but less than 1,000 parts by mass, even more preferably 20 parts by mass or more but less than 500 parts by mass, even more preferably 30 parts by mass or more but less than 300 parts by mass, even more preferably 30 parts by mass or more but less than 250 parts by mass, and particularly preferably 30 parts by mass or more but less than 200 parts by mass. When the content of the active ingredient (B) per 100 parts by mass of the hyaluronic acid derivative (A) is below the upper limit, the active ingredient becomes amorphous, improving the dissolution rate of the active ingredient. On the other hand, when the content is above the lower limit, the contact area between the active ingredient and water increases, improving the dissolution rate of the active ingredient. Here, the dissolution rate of the active ingredient (B) in the formulation can be verified and the ratio can be optimized using the dissolution test method specified by the Japanese Pharmacopoeia First Fluid (JP1) or Japanese Pharmacopoeia Second Fluid (JP2).

[0113] The active ingredient (B) may be formulated as a single ingredient, or as a combination of two or more ingredients.

[0114] The pharmaceutical composition of this embodiment can solubilize the active ingredient (B) without using an organic solvent when formulating the active ingredient, thereby reducing the need for conventional, highly toxic solubilizing agents. Note that the desired effect may be achieved by a mechanism different from the above-mentioned mechanism. In other words, the pharmaceutical composition of a hyaluronic acid derivative for oral absorption of this embodiment can also be said to be a composition that does not contain organic solvents.

[0115] <Other additives> The pharmaceutical composition of this embodiment may consist solely of (A) a hyaluronic acid derivative and (B) an active ingredient, or may contain other additives in addition to these. The other additives are not particularly limited as long as they are pharmacologically acceptable, and may be selected appropriately depending on the dosage form. Examples of such additives include water or other physiologically acceptable liquids (e.g., saline, phosphate-buffered saline (PBS)), vehicles, buffers, surfactants, pH adjusters, isotonicity agents, thickeners, preservatives, stabilizers, cryoprotectants, excipients, binders, disintegrants, and polymers commonly used in pharmaceuticals. These additives may be used alone or in combination of two or more of the same or different additives.

[0116] Examples of buffering agents include Tris, sodium citrate hydrate, sodium acetate hydrate, sodium bicarbonate, dry sodium carbonate, sodium carbonate, magnesium sulfate, trometamol, boric acid, borax, sodium hydrogen phosphate hydrate, sodium dihydrogen phosphate, potassium dihydrogen phosphate, and histidine.

[0117] The surfactant can also be used as an association promoter for the hyaluronic acid derivative (A). As the association promoter, a surfactant containing at least four ether structures (ROR) and having four or more carbon atoms is preferred. Examples of the surfactant include polysorbate (having five or more ether structures and having ten or more carbon atoms), polyoxyethylene fatty acid ester, sorbitan fatty acid ester, polyethylene castor oil, etc.

[0118] Examples of pH adjusters include hydrochloric acid and sodium hydroxide.

[0119] Examples of isotonic agents include potassium chloride, calcium chloride, sodium chloride, concentrated glycerin, glucose, D-mannitol, and the like.

[0120] Examples of thickening agents include carboxyvinyl polymer, povidone, polyvinyl alcohol (partially saponified), hydroxyethyl cellulose, hydroxypropyl cellulose, hydroxypropylmethyl cellulose, hypromellose, methyl cellulose, and glycerin.

[0121] Examples of preservatives include benzalkonium chloride, methyl parahydroxybenzoate, propyl parahydroxybenzoate, chlorobutanol, sorbic acid, and alkylpolyaminoethylglycine.

[0122] Examples of stabilizers include sodium edetate hydrate and polyvinylpyrrolidone (povidone).

[0123] The cryoprotectant is not particularly limited as long as it is known as a "cryoprotectant" or "lyoprotectant," and examples thereof include disaccharides, sorbitol, dextran, polyethylene glycol, propylene glycol, glycerin, glycerol, polyvinylpyrrolidone, dimethyl sulfoxide, etc.

[0124] Disaccharide is not particularly limited, and for example, can be 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, primeverose etc.Among them, sucrose, trehalose, maltose or lactose are preferred, because they are widely used as cryoprotective agent.In addition, sucrose is more preferred, because of its use as pharmaceutical additive and from the viewpoint of more effectively suppressing the increase in particle size of the microparticles formed by hyaluronic acid derivative during freeze-drying.

[0125] Examples of excipients include cellulose, starch acrylate, L-aspartic acid, aminoethylsulfonic acid, aminoacetic acid, candy (powder), gum arabic, powdered gum arabic, alginic acid, sodium alginate, pregelatinized starch, pumice granules, inositol, ethylcellulose, ethylene vinyl acetate copolymer, sodium chloride, olive oil, kaolin, cacao butter, casein, fructose, pumice granules, carmellose, carmellose sodium, hydrated silicon dioxide, dry yeast, dried aluminum hydroxide gel, dried sodium sulfate, dried magnesium sulfate, agar, and kaolin. Ingredients: cetethane powder, citric acid, sodium citrate, disodium citrate, glycerin, calcium glycerophosphate, sodium gluconate, L-glutamine, clay, clay 3, clay granules, croscarmellose sodium, crospovidone, magnesium aluminosilicate, calcium silicate, magnesium silicate, light anhydrous silicic acid, light liquid paraffin, cinnamon powder, genmai koji, synthetic aluminum silicate, synthetic hydrotalcite, sesame oil, wheat flour, wheat germ flour, rice flour, rice starch, potassium acetate, calcium acetate, cellulose acetate phthalate, safflower Ingredients: oil, white beeswax, zinc oxide, titanium oxide, magnesium oxide, β-cyclodextrin, dihydroxyaluminum aminoacetate, 2,6-di-butyl-4-methylphenol, dimethylpolysiloxane, tartaric acid, potassium hydrogen tartrate, calcined gypsum, sucrose fatty acid ester, magnesium alumina hydroxide, aluminum hydroxide gel, aluminum hydroxide-sodium bicarbonate coprecipitate, magnesium hydroxide, squalane, stearyl alcohol, stearic acid, calcium stearate, polyoxyethylene stearate, magnesium stearate, dimethicone Hardened oil, refined gelatin, refined shellac, refined white sugar, refined white sugar spherical granules, cetostearyl alcohol, polyethylene glycol 1000 monocetyl ether, gelatin, sorbitan fatty acid ester, tricalcium phosphate, soybean oil, soybean unsaponifiables, soybean lecithin, skim milk powder, talc, ammonium carbonate, calcium carbonate, magnesium carbonate, neutral anhydrous sodium sulfate, low-substituted hydroxypropyl cellulose, dextran, dextrin, natural aluminum silicate, tragacanth powder, silicon dioxide, calcium lactate, Perfiller 101, white shellac,White petrolatum, Hakudo, white sugar, white sugar and starch spherical granules, naked barley leaf extract powder, dried naked malt leaf green juice powder, honey, paraffin, potato starch, semi-digested starch, human serum albumin, hydroxypropyl starch, hydroxypropyl cellulose, hydroxypropyl methylcellulose phthalate, hydroxypropyl methylcellulose phthalate, phytic acid, glucose, glucose hydrate, partially pregelatinized starch, pullulan, propylene glycol, powdered reduced maltose syrup, pectin, bentonite, sodium polyacrylate, polyoxyethylene alkyl ether, polyoxyethylene hydrogenated castor oil, polyoxyethylene (105) polyoxypropylene (5) glycol, polyoxyethylene (160) polyoxypropylene (30) glycol, polystyrene Examples of suitable soluble calcium carbonates include sodium phosphate, polysorbate 80, polyvinyl acetal diethylaminoacetate, polyvinylpyrrolidone, polyethylene glycol, maltose, starch syrup, isopropyl myristate, anhydrous lactose, anhydrous calcium hydrogen phosphate, anhydrous calcium phosphate granules, magnesium aluminometasilicate, methylcellulose, cottonseed flour, cottonseed oil, Japan wax, aluminum monostearate, glycerin monostearate, sorbitan monostearate, medicinal charcoal, peanut oil, aluminum sulfate, calcium sulfate, granular cornstarch starch, liquid paraffin, dl-malic acid, calcium hydrogen phosphate, calcium hydrogen phosphate, calcium hydrogen phosphate granules, sodium hydrogen phosphate, potassium dihydrogen phosphate, calcium dihydrogen phosphate, and sodium dihydrogen phosphate.

[0126] Examples of binders include sugars such as sucrose, fructose, lactose or lactose hydrate, fructooligosaccharides, glucose, palatinose, maltose, reduced maltose, powdered sugar, powdered candy, isomerized lactose, and honey sugar; sugar alcohols such as mannitol, xylitol, maltitol, erythritol, sorbitol, and lactitol; water-soluble polysaccharides such as gelatin, pullulan, carrageenan, locust bean gum, agar, glucomannan, xanthan gum, tamarind gum, pectin, sodium alginate, and gum arabic; celluloses such as hydroxypropyl cellulose and methyl cellulose; starches such as corn starch, potato starch, rice starch, pregelatinized starch, and starch paste; synthetic polymers such as polyvinylpyrrolidone, carboxyvinyl polymer, and polyvinyl alcohol; and inorganic compounds such as calcium hydrogen phosphate, calcium carbonate, synthetic hydrotalcite, and magnesium aluminosilicate.

[0127] Examples of disintegrants include celluloses such as croscarmellose sodium, carmellose, carmellose calcium, carmellose sodium, and low-substituted hydroxypropyl cellulose; starches such as carboxymethyl starch sodium, hydroxypropyl starch, rice starch, wheat starch, potato starch, pregelatinized starch, and partially pregelatinized starch; and synthetic polymers such as crospovidone and crospovidone copolymer.

[0128] Polymers generally used as pharmaceuticals are not particularly limited, and examples thereof include nonionic cellulose-based polymers, ionic cellulose-based polymers, synthetic polymer monopolymers, and synthetic polymer copolymers.

[0129] Examples of nonionic cellulose polymers include hypromellose (manufactured by Shin-Etsu Chemical Co., Ltd.: TC-5E, TC-5R, Metolose 60SH, etc.), hydroxypropyl cellulose (manufactured by Nippon Soda Co., Ltd.: NISSO HPC-L, HPC-SSL, etc.), hydroxyethyl methylcellulose, hydroxypropyl methylcellulose acetate, and hydroxyethyl cellulose acetate.

[0130] Examples of ionic cellulose polymers include hydroxypropyl methylcellulose acetate succinate (Shin-Etsu Chemical Co., Ltd.: Shin-Etsu AQOAT-AS-LF, AS-MF, AS-HG, etc.), hydroxypropyl methylcellulose succinate, hydroxypropyl cellulose acetate succinate, hydroxyethyl methylcellulose succinate, hydroxyethyl cellulose acetate succinate, hydroxypropyl methylcellulose phthalate (Shin-Etsu Chemical Co., Ltd.: HP-50, HP-55, etc.), hydroxyethyl methylcellulose acetate succinate, hydroxyethyl methylcellulose acetate phthalate, carboxyethyl cellulose, ethyl carboxymethyl cellulose, carboxymethyl cellulose, cellulose acetate phthalate, methyl cellulose acetate phthalate, ethyl cellulose acetate phthalate, hydroxypropyl cellulose acetate phthalate, hydroxypropyl methylcellulose acetate phthalate, hydroxypropyl cellulose acetate phthalate succin ... phthalate, hydroxypropyl cellulose acetate phthalate succinate, hydroxypropyl methylcellulose phthalate, hydroxypropyl cellulose acetate phthalate succinate, hydroxypropyl methylcellulose phthalate, hydroxypropyl cellulose acetate phthalate succinate, hydroxypropyl methylcellulose phthalate, hydroxypropyl cellulose acetate phthalate succinate, hydroxypropyl methylcellulose phthalate, hydroxypropyl cellulose acetate phthalate succinate, hydroxypropyl methylcellulose phthalate, hydroxypropyl cellulose acetate phthalate succ Hydroxypropyl methylcellulose acetate succinate phthalate, hydroxypropyl methylcellulose succinate phthalate, cellulose propionate phthalate, hydroxypropyl cellulose butyrate phthalate, cellulose acetate trimellitate, methylcellulose acetate trimellitate, ethylcellulose acetate trimellitate, hydroxypropyl cellulose acetate trimellitate, hydroxypropyl methylcellulose acetate trimellitate, hydroxypropyl cellulose acetate trimellitate succinate cellulose acetate, cellulose propionate trimellitate, cellulose butyrate trimellitate, cellulose acetate terephthalate, cellulose acetate isophthalate, cellulose acetate pyridine carboxylate, salicylic acid cellulose acetate, hydroxypropyl salicylic acid cellulose acetate, ethyl benzoic acid cellulose acetate, hydroxypropyl ethyl benzoic acid cellulose acetate, ethyl phthalic acid cellulose acetate, ethyl nicotinic acid cellulose acetate, and ethyl picolinic acid cellulose acetate.

[0131] Examples of synthetic polymer monopolymers include polyvinylpyrrolidone (manufactured by BASF: Kollodon K30, K90, K17, etc.), crosslinked polyvinylpyrrolidone, polyethylene glycol (manufactured by Sanyo Chemical Industries, Ltd.: Macrogol 4000, 6000, 20000, etc.), polyvinyl alcohol, polyvinyl acetate phthalate, polyvinyl acetal diethylaminoacetate, and carboxyvinyl polymer. Examples of synthetic polymer copolymers include methacrylic acid copolymers (manufactured by Rohm: Eudragit L100, L100-55, S100, L30D-55, etc.), aminoalkyl methacrylate copolymers (manufactured by Rohm: Eudragit E100, RS100, etc.), polyvinyl alcohol polyvinyl acetate copolymers, polyethylene glycol polypropylene glycol copolymers, polyethylene polyvinyl alcohol copolymers, and polyvinylpyrrolidone copolymers (manufactured by BASF: Kollodon VA64, SR, etc.).

[0132] Of the above polymers, preferred are hypromellose, hydroxypropyl cellulose, hydroxypropyl methylcellulose phthalate, hydroxypropyl methylcellulose acetate succinate, polyvinylpyrrolidone, polyvinylpyrrolidone copolymers, and methacrylic acid copolymers.

[0133] Furthermore, among polymers generally used in pharmaceuticals, examples of polymers used as coating agents for providing sustained release to active ingredients include gum arabic, agar, guar gum, cereal gum, dextran, casein, pectin, carrageenan, wax, shellac, hydrogenated vegetable oil, hydroxypropyl cellulose (Nippon Soda: HPC-SSL, -SLL, etc.), hydroxyethyl cellulose (HEC), hydroxypropyl methylcellulose (Shin-Etsu Chemical: TC-5 series, etc.), sodium carboxymethylcellulose, poly(ethylene) oxide, alkyl cellulose, ethyl cellulose (Dow Chemical: Ethocel, etc.), methyl cellulose (Shin-Etsu Chemical: Metrose, etc.), and the like. SM-25, -100, etc.), carboxymethyl cellulose (CMC), carboxymethyl ethyl cellulose (Freund Corporation: CMEC), polyethylene glycol, polyvinylpyrrolidone, cellulose acetate, cellulose acetate butyrate, cellulose acetate phthalate, cellulose acetate trimellitate, polyvinyl acetate phthalate, hydroxypropyl methylcellulose phthalate, hydroxypropyl methylcellulose acetate succinate, polyacetal diethylaminovinyl acetate, poly(alkyl methacrylate), poly(vinyl acetate), acrylic acid or methacrylic acid and the like Examples of suitable copolymers include polymers derived from the respective esters of acrylic acid or methacrylic acid, copolymers derived from acrylic acid or methacrylic acid and the respective esters (e.g., Eudragit RL100, RLP0, RL30D, RS100, RSP0, RS30D, NE30D, E100, EPO, L100, L100-55, L30D-55, S100, etc., manufactured by Rohm), polyvinyl alcohol polyethylene glycol graft copolymer (Kollicoat IR, manufactured by BASF), and polyvinyl acetal diethylaminoacetate (AEA, manufactured by Mitsubishi Chemical Foods). Preferred are ethyl cellulose, Eudragit, CMEC, AEA, and Kollicoat IR.

[0134] A polymer that is generally used as a pharmaceutical may be added to the hyaluronic acid derivative beforehand to prepare a solid dispersion, or may be added after preparing a solid dispersion with the hyaluronic acid derivative.

[0135] ≪Form≫ The pharmaceutical composition of this embodiment is a pharmaceutical composition intended to be absorbed in the stomach or intestinal tract during oral administration, and the dosage form is not particularly limited as long as it is absorbable in the stomach or intestinal tract. For example, the pharmaceutical composition of this embodiment can be formulated into a dosage form suitable for oral administration or enteral administration. Enteral administration can be via nasal administration, gastrostomy, enterostomy, etc.

[0136] The dosage form of the pharmaceutical composition of this embodiment may be any dosage form that allows oral administration or tube administration, and may be in the form of a solid, semi-solid, or liquid. In the case of solids, examples include powders, granules, pills, pellets, tablets, capsules, etc. Among these, as solids, tablets and capsule preparations in which freeze-dried powders or granules thereof are coated with an enteric coating agent are preferred. When aiming for local retention and absorption in the stomach, it is more preferable not to use an enteric coating agent. In the case of a semi-solid, examples include a gel and the like. In the case of a liquid, examples include a suspension in which the powder is diluted or suspended in water or a buffer solution such as phosphate buffer (PB) or phosphate buffered saline (PBS).

[0137] In the pharmaceutical composition of this embodiment, when (A) the hyaluronic acid derivative and (B) the active ingredient form nanoparticles, the pharmaceutical composition of this embodiment can be a suspension, granules, tablets, or capsule formulation. For example, the nanoparticles can be dispersed in water or other physiologically acceptable liquid to form a suspension, or the nanoparticles can be freeze-dried in the presence of a cryoprotectant to form a powder, which can be used as a raw material to form granules, tablets, or capsule formulations. In the pharmaceutical composition of this embodiment, when (A) the hyaluronic acid derivative and (B) the active ingredient form a solid dispersion, the pharmaceutical composition of this embodiment can be a solution, granules, tablets, or capsule formulations. For example, the solid dispersion can be dispersed or dissolved in water or other physiologically acceptable liquid to form a liquid, or the dry powder of the solid dispersion can be used as a raw material to form granules, tablets, or capsule formulations.

[0138] Capsules can be obtained by filling solid dispersions as they are, solid dispersions with additives, or solid dispersions with additives and granulated. Furthermore, lubricants and other additives may be added as needed before filling capsules. Lubricants may be added to impart fluidity to the powder and prevent mechanical adhesion during capsule filling. Other examples include microcapsules prepared by coacervation techniques or interfacial polymerization, such as hydroxymethylcellulose microcapsules, gelatin microcapsules, or poly(methyl methacrylate) microcapsules. Examples include push-fit capsules made of gelatin ("gelcaps") and soft, sealed capsules containing gelatin and a plasticizer such as glycerol or sorbitol. Push-fit capsules can contain fillers such as lactose, binders such as starch, and / or lubricants such as talc or magnesium stearate, and can optionally contain the active ingredient in a mixture with stabilizers. In soft capsules, the active compound can be dissolved or suspended in a suitable liquid, such as fatty oils, liquid paraffin, or liquid polyethylene glycol. Suitably, capsules will typically be up to 1 gram in size, for example from about 0.01 grams to about 0.8 grams in size.

[0139] Tablets can have a variety of shapes, including diamond, modified capsule, modified oval, and hexagonal, and can optionally have beveled surfaces. The specific type and amount of excipients and the selection of tableting techniques depend on the excipient's additional properties, such as compressibility, flowability, particle size, compatibility, and density. Tablets can be prepared according to methods known in the art, including direct compression, dry granulation, fluidized bed granulation, and wet granulation. Therefore, the type of excipients used varies. Wet granulation is known to be particularly suitable for providing high strength, low breakage, and suitable for commercial manufacturing scale.Suitable wet granulation tablets include granules that contain one or more fillers, binders and disintegrants.The granules are mixed with additional fillers, binders, disintegrants and / or lubricants to form a compression mixture that is compressed to form tablets.

[0140] <Method for manufacturing nanoparticles> For example, by mixing a solution (solution (A)) containing a hyaluronic acid derivative (A) with a solution (solution (B)) containing an active ingredient (B), the hyaluronic acid derivative (A) and the active ingredient (B) can form nanoparticles. Solution (A) can be prepared by dissolving the hyaluronic acid derivative (A) in water, phosphate buffer (PB), phosphate-buffered saline (PBS), or the like. Solution (B) can be prepared by dissolving the active ingredient (B) in water to which various organic solvents or solubilizing agents such as vehicles have been added. Simply mixing and stirring the two solutions can result in the formation of nanoparticles of the hyaluronic acid derivative (A) and the active ingredient (B) through interactions between the hydrophobic moieties, such as steryl groups, in the hyaluronic acid derivative (A) and the hydrophobic moieties in the active ingredient (B), or through self-assembly of the hyaluronic acid derivatives themselves.

[0141] The complex formed in the solution can also be dried and powdered.Drying methods include, for example, ventilation drying, drying in a thermostatic oven, drying under reduced pressure, hot air circulation drying, freeze-drying, etc.Among them, freeze-drying is preferred.When the complex of (A) hyaluronic acid derivative and (B) active ingredient is in the form of nanoparticles, it is preferred to further include a cryoprotectant when freeze-drying.

[0142] 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 inside the apparatus during freeze-drying is preferred, and from the viewpoint of controlling the product temperature, a tray-type freeze-dryer is preferred.

[0143] <Method for producing solid dispersion> The solid dispersion used in this embodiment can be produced by a method generally used to produce a solid dispersion of a drug, such as a solvent method, a melting method, or a mechanochemical method, with the solvent method being preferred. In the solvent method, (A) a hyaluronic acid derivative, (B) an active ingredient, and, if necessary, other pharmaceutical additives are suspended or dissolved in an organic solvent, and then the solvent is removed or the (A) hyaluronic acid derivative and (B) an active ingredient are precipitated to produce the drug.

[0144] Examples of the solvent method include evaporation, spraying, coprecipitation, filtration, freeze-drying, etc. Examples of the spraying method include fluidized bed, spray-drying, tumbling bed, stirring, and supercritical methods.

[0145] Among them, spraying method is preferred, because it can remove the solvent in a short time, and can obtain the solid dispersion that (A) hyaluronic acid derivative and (B) active ingredient are in the same molecular dispersion state as when they are in the solvent.Among the spraying methods, spray drying method is preferred, because it can remove the solvent instantly, can be mass-produced continuously, and can easily obtain the desired powder properties such as particle size and bulk density.

[0146] The time required for removing the solvent is preferably short, preferably within 120 minutes, more preferably within 60 minutes, even more preferably within 10 minutes, particularly preferably within 5 minutes, and most preferably within 2 minutes. By keeping the time within the upper limit, the degree of molecular dispersion of (A) the hyaluronic acid derivative and (B) the active ingredient is high. On the other hand, if the time required to form the solid dispersion is long, fine crystals of the (B) active ingredient and polymer pores are likely to form. The time required to remove the solvent here refers to the time required to obtain a solid solid dispersion, and some of the solvent may remain. The remaining solvent can be removed by secondary drying, which will be described later.

[0147] The solvent used in the solvent method may be any pharmaceutically acceptable solvent, and examples thereof include (B) solvents in which the active ingredient dissolves, such as ethanol, methanol, 2-propanol, acetone, 2-butanone, methyl isobutyl ketone, tetrahydrofuran (THF), tetrahydropyran, 1,4-dioxane, diethyl ether, diisopropyl ether, t-butyl methyl ether, hexane, heptane, toluene, acetonitrile, methylene chloride, chloroform, carbon tetrachloride, methyl acetate, ethyl acetate, butyl acetate, acetic acid, formic acid, N,N-dimethylformamide (DMF), N,N-dimethylacetamide (DMAc), dimethyl sulfoxide (DMSO), and water. These can be used alone or as a mixed solvent of two or more types depending on the properties of the desired solid dispersion.

[0148] Among the solvent methods, the coprecipitation method is a method for precipitating (A) hyaluronic acid derivatives and (B) active ingredients. In the coprecipitation method, the (A) hyaluronic acid derivatives and (B) active ingredients are dissolved or suspended in a solvent, and then a solvent in which either or both of the (A) hyaluronic acid derivative and the (B) active ingredient are insoluble is added, or the temperature of the solvent is lowered to reduce the solubility, thereby obtaining a solid dispersion of the (A) hyaluronic acid derivatives and the (B) active ingredient. As an example of the coprecipitation method, the (A) hyaluronic acid derivatives and the (B) active ingredient are dissolved in a solvent, and then the solution is added dropwise or reversely dropped into a medium with a high salt concentration, such as 10xPBS (phosphate buffer containing 1500 mM NaCl, manufactured by Fujifilm Wako Co., Ltd., product number: ), thereby precipitating the (A) hyaluronic acid derivatives and the (B) active ingredient, thereby obtaining a solid dispersion of the (B) active ingredient. In this case, divalent salts such as CaCl2 and MgCl2 may be used, or precipitation may be performed using a trivalent salt.

[0149] The evaporation method is described in detail below. (A) A hyaluronic acid derivative and (B) an active ingredient are dissolved or suspended in a solvent. The solids concentration at this time is 0.2 to 40% by weight, preferably 1 to 20% by weight. The solvent is then removed under reduced pressure or normal pressure. The temperature at this time is selected appropriately based on the distillation time and allows the solvent to be distilled off, and is -10 to 120°C, preferably 0 to 100°C.

[0150] The production method using the spray drying method is described in detail below. (A) A hyaluronic acid derivative and (B) an active ingredient are dissolved or suspended in an organic solvent. The concentration at this time may be any concentration that allows spray drying, with a solid content of 0.1 to 80% by weight, preferably 0.5 to 50% by weight. Next, solvent removal and granulation are simultaneously carried out by spray drying. A disk-type or nozzle-type spray dryer (e.g., a pressure nozzle, a two-fluid nozzle, or a four-fluid nozzle) is used as the spray dryer. The temperatures during spray drying are preferably an inlet temperature of approximately 20 to 150°C and an outlet temperature of approximately 0 to 85°C.

[0151] If further removal of residual solvent is required after obtaining a solid dispersion by a solvent method such as evaporation or spray drying, secondary drying can be performed. The secondary drying can be performed by any drying method commonly used in pharmaceutical manufacturing, as long as it can maintain the stability of the poorly soluble drug solid dispersion.

[0152] The melting method is a method for producing a solid dispersion by heating (A) a hyaluronic acid derivative, (B) an active ingredient, and, if necessary, other pharmaceutical additives to a temperature above the melting point of either ingredient, melting them, and then cooling them. The method can be selected arbitrarily based on the equipment, temperature, and conventional methods used. Examples of equipment include extruders. A solvent may be added to lower the melting point, and the solvent is removed after melting.

[0153] The mechanochemical method is a method for producing a solid dispersion by simultaneously grinding and impacting (A) a hyaluronic acid derivative and (B) an active ingredient, and optionally other pharmaceutical additives. The machines, temperatures, and grinding conditions used can be selected as desired according to conventional methods.

[0154] The solid dispersion thus obtained can be subjected to dry granulation or wet granulation using pharmaceutical additives for tablet mixability, dissolution, sustained release, and bitterness masking. If the solid dispersion has poor mixability with pharmaceutical additives due to differences in particle size or bulk density, the particle size or bulk density can be increased by densifying the solid dispersion using a roller compactor or the like.

[0155] Pharmaceutical additives are mixed with either or both of the solid dispersion and the solid dispersion-containing particles to obtain oral pharmaceutical compositions such as tablets, capsules, powders, liquids, emulsions, or suspensions. Pharmaceutical compositions for parenteral use, such as injections, suppositories, eye drops, inhalants, and topical skin preparations, can also be obtained. These can be produced by known methods.

[0156] <Administration method> The pharmaceutical composition of this embodiment can be administered to animals classified as mammals, including humans (monkeys, marmosets, mice, rats, cows, horses, cats, dogs, pigs, sheep, goats, rabbits, etc.).

[0157] The administration route of the pharmaceutical composition of this embodiment is not particularly limited as long as it is an administration route that allows the active ingredient (B) to be absorbed from the intestinal tract, and can be appropriately selected depending on the intended use, the location of the tissue to be treated, etc. The administration route of the pharmaceutical composition of this embodiment is preferably oral administration, but may also be enteral administration via a gastrostomy, enterostomy, or nasogastric tube.

[0158] The dosage of the pharmaceutical composition of this embodiment can be appropriately selected taking into consideration the type of subject (including age, sex, etc.). Generally, for example, for a human (body weight 60 kg), the daily dosage is, in terms of the amount of (B) active ingredient (preferably, a protein, peptide, or low-molecular-weight compound), preferably 1 mg to 1000 mg, more preferably 10 mg to 500 mg; for example, 15, 20, 25, 50, 75, or 100 mg, of the active ingredient, when the unit dosage form is, for example, a tablet, capsule, or powder. Such a unit dosage form is suitable for administration once to five times daily, depending on the purpose of treatment, the progress of treatment, and other factors.

[0159] The administration frequency may be a single administration of the above-mentioned dosage, or multiple administrations of the above-mentioned dosage, such as once every day, every two days, four days, one week, two weeks, three weeks, four weeks, one month, two months, three months, or six months, etc. Alternatively, administration may be performed at two or more sites in a single administration.

[0160] Other Embodiments In one embodiment, the present invention provides a method for preventing or treating a disease, which comprises administering an effective amount of the pharmaceutical composition to a human or animal patient. Furthermore, the term "effective amount" as used herein includes an amount effective for prevention or treatment, that is, an amount suitable for preventing the onset or treatment of the above-mentioned diseases.

[0161] In one embodiment, the present invention provides a composition for the prevention or treatment of one or more diseases selected from the group consisting of cancer, infectious diseases, immune diseases, inflammatory diseases, allergic diseases, skin diseases, hypertension, diabetes, neurological diseases, genetic diseases, cardiovascular diseases, cerebrovascular diseases, respiratory diseases, eye diseases, ear diseases, and bone and joint diseases, the composition comprising the above-mentioned active ingredient-hyaluronic acid derivative complex.

[0162] In one embodiment, the present invention provides use of the above-mentioned active ingredient-hyaluronic acid derivative complex for the manufacture of a pharmaceutical composition.

[0163] In the pharmaceutical composition of this embodiment, the (A) hyaluronic acid derivative interacts with CD44-positive cells (hyaluronic acid receptors) present in the intestinal tract at absorption sites in the stomach, small intestine (duodenum, jejunum, ileum), and large intestine, or contributes to retention due to viscosity, thereby increasing the local concentration of the active ingredient in the intestinal tract, thereby potentially enhancing local medicinal efficacy. Therefore, the pharmaceutical composition of the present application may also be useful for diseases of the stomach, small intestine, and large intestine. Furthermore, the (A) hyaluronic acid derivative may contribute to inhibiting the degradation of the (B) active ingredient in the acidic environment of the stomach. Additionally, crystallization of the (B) active ingredient due to pH changes during its transition from the stomach to the intestine is often a problem, but the (A) hyaluronic acid derivative may protect against this.

[0164] Examples of gastrointestinal diseases include dyspepsia, gastroparesis, postoperative gastric ileus, functional esophageal disorder, functional gastroduodenal disorder, gastroesophageal reflux disease (GERD), gastroduodenal ulcer, and gastric ulcer. Examples of intestinal diseases include inflammatory bowel disease, representative examples of which include ulcerative colitis and irritable bowel syndrome, irritable bowel syndrome and Crohn's disease, celiac disease, GI-GVHD, gastroenteritis, duodenitis, jejunitis, ileitis, peptic ulcer, curling ulcer, appendicitis, colitis, pseudomembranous colitis, irritable bowel syndrome (irritable bowel syndrome-diarrhea predominant (IBS-D), irritable bowel syndrome-constipation predominant (IBS-C), and irritable bowel syndrome-mixed predominant (IBS-M)), multiple diverticulosis, diverticulitis, and endometriosis. [Example]

[0165] The present invention will be described in detail below with reference to examples, but it is not intended that the scope of the present invention be limited to these examples.

[0166] <Synthesis of hyaluronic acid derivatives> [Synthesis Example 1] As described in Examples 1 and 2 of International Publication No. 2010 / 053140, a hyaluronic acid derivative with cholesteryl groups introduced (weight-average molecular weight of 10 kDa, cholesteryl group introduction rate of 30%) (hereinafter, sometimes referred to as "CHHA10k30") was synthesized.

[0167] [Synthesis Example 2] As described in Examples 1 and 2 of International Publication No. 2010 / 053140, a hyaluronic acid derivative with cholesteryl groups introduced (weight-average molecular weight of 10 kDa, cholesteryl group introduction rate of 40%) (hereinafter, sometimes referred to as "CHHA10k40") was synthesized.

[0168] <Test Example 1> In Test Example 1, a formulation of cyclosporine (CyA), a poorly water-soluble peptide, was prepared.

[0169] [Comparative Example 1-1] A solution was prepared in which CyA was dissolved using polyethylene glycol 400 (PEG400) and Cremophor EL (CAS. RN: 61791-12-6) (hereinafter sometimes referred to as "CreEL") as a solubilizing agent.

[0170] The following operations were carried out at room temperature (20°C). CyA powder (Tokyo Chemical Industry Co., Ltd., product number: C2408) was dissolved at 1.0 mg / mL by adding DMSO, PEG400, CreEL, and PBS in that order. Specifically, a solution formulation was prepared using a DMSO / PEG400 / CreEL / PBS (volume ratio: 5:30:2.5:62.5) solution. The solution was visually clear. The final formulation composition is shown in Table 1.

[0171] [Example 1-1] A solution formulation using a hyaluronic acid derivative (10k HA-C6-Chol-30%) was prepared. The following operations were carried out at room temperature (20°C). The freeze-dried hyaluronic acid derivative (10k HA-C6-Chol-30%) obtained in Synthesis Example 1 was dissolved in a 10% sucrose solution by stirring for 24 hours to an isotonic solution at 10.0 mg / mL. 9.0 mg of powdered CyA (Tokyo Chemical Industry Co., Ltd., product number: C2408) was weighed into a separate vial. Next, 9.0 mL of 10 mg / mL hyaluronic acid derivative aqueous solution was added to the vial containing the powder, and the drug was complexed while stirring with a stirrer.Furthermore, the hyaluronic acid derivative aqueous solution was added until the drug residue was almost completely dissolved by visual inspection, and then the mixture was stirred for 12 hours to homogenize, and the pharmaceutical composition was prepared.The solution was visually clear.The final formulation composition is shown in Table 1.

[0172] [Example 1-2] A solution formulation using a hyaluronic acid derivative (10k HA-C6-Chol-40%) was prepared. A solution formulation using a hyaluronic acid derivative (10k HA-C6-Chol-40%) was prepared in the same manner as in Example 1-1, except that the freeze-dried hyaluronic acid derivative (10k HA-C6-Chol-40%) obtained in Synthesis Example 2 was used. The solution was visually clear. The final formulation composition is shown in Table 1.

[0173] [Comparative Example 1-2] A solution formulation using the hyaluronic acid derivative (10k HA-C6-Chol-40%) was prepared in the same manner as in Example 1-2, except that the freeze-dried hyaluronic acid derivative (10k HA-C6-Chol-40%) obtained in Synthesis Example 2 was dissolved in 10% sucrose aqueous solution at 50.0 mg / mL. The solution was visually clear. The final formulation composition is shown in Table 1.

[0174] [Table 1]

[0175] [Comparative Example 1-3] A nanocrystal formulation of CyA was prepared. The following operations were carried out at room temperature (20°C). 720 mg of CyA (Tokyo Chemical Industry Co., Ltd., product number: C2408) was weighed into a 30 mL glass vial and 14.4 mL of 0.01% polysorbate 80 was added. 10 g of 0.2 mm ZrO2 beads (Nikkato Corporation) and 14.4 mL of 1% polysorbate 80 were then added. A rotor was added and the mixture was stirred with a magnetic stirrer for 11 days to finely pulverize the mixture. The CyA suspension was then removed with a syringe and placed in a 15 mL conical tube. The tube was centrifuged at 10,000 g for 60 minutes and the supernatant was discarded. Ten volumes of water for injection were then added, and the tube was centrifuged at 10,000 g for 60 minutes. The supernatant was discarded. The resulting cyclosporine suspension was vacuum-dried to prepare a CyA nanocrystal formulation. The final formulation composition is shown in Table 2.

[0176] [Examples 1-3] A solid formulation using a hyaluronic acid derivative (10k HA-C6-Chol-40%) was prepared. The following operations were carried out at room temperature (20°C). Hyaluronic acid derivative (10k HA-C6-Chol-40%) was dissolved in DMSO solution at 25.0 mg / mL by stirring at 75°C for 2 hours. 100 mg of powdered CyA (Tokyo Chemical Industry Co., Ltd., product number C2408) was weighed into a separate vial. It was then dissolved in DMSO solution at room temperature at 50 mg / mL. Next, 4000 μL of the 25 mg / mL hyaluronic acid derivative-containing DMSO solution was added to another vial, and 800 μL of the cyclosporine-containing DMSO solution was added to the other vial. After thorough vortex mixing, the mixture was freeze-dried. The resulting freeze-dried powder was analyzed by XRD.

[0177] (XRD measurement conditions) Measurement equipment: Rigaku Ultima-IV X-ray source: Cu-Kα Excitation voltage: 40 kV, current 40 mA Optical system: Concentrated optical system Cu-Kβ ray filter: Ni foil Absorber: None Detector: Dtex (high sensitivity detector) Measurement method: θ / 2θ method Slit: DS=1°, SS=open, RS=open, vertical slit=10mm 2θ / θ scan: 2θ=5~75° (0.02° / step, 10° / min)

[0178] As shown in Figure 1, the results of XRD analysis showed that no crystalline peaks of cyclosporine were observed. In other words, a solid formulation consisting of a pharmaceutical composition, which is a solid dispersion of cyclosporine in an amorphous state, was successfully prepared. The final formulation composition is shown in Table 2.

[0179] [Examples 1-4] A solid preparation using hyaluronic acid derivative (10k HA-C6-Chol-30%) was prepared. Except that the hyaluronic acid derivative (10k HA-C6-Chol-30%) was dissolved in DMSO solution at 25.0mg / mL, it was prepared in the same manner as the method described in Example 1-3, and XRD analysis was carried out.

[0180] As shown in Figure 2, the XRD analysis results confirmed that cyclosporine was amorphous, as in Examples 1-3. That is, a solid formulation consisting of a pharmaceutical composition, which is a solid dispersion in which cyclosporine is maintained in an amorphous state, was successfully prepared. The final formulation composition is shown in Table 2.

[0181] [Examples 1-5] A solid formulation using a hyaluronic acid derivative (10k HA-C6-Chol-30%) was prepared. The hyaluronic acid derivative (10k HA-C6-Chol-30%) was dissolved in DMSO solution at 25.0 mg / mL. 100 mg of powdered CyA (Tokyo Chemical Industry Co., Ltd., product number C2408) was weighed into a separate vial. It was then dissolved in DMSO solution at 50 mg / mL at room temperature. Next, 3000 μL of the 25 mg / mL hyaluronic acid derivative-containing DMSO solution was added to another vial, and 600 μL of the CyA-containing DMSO solution was added to the other vial. The mixture was thoroughly vortexed to homogenize. 3.6 mL of the resulting mixture was added dropwise to 30 mL of 10x PBS and allowed to stand at room temperature for 24 hours, resulting in a precipitate of a complex of the hyaluronic acid derivative and CyA. The mixture was then centrifuged at 10,000 × g for 5 minutes at 4°C to settle the precipitate, and the supernatant was removed. The precipitate was washed with 10 × PBS, then with 1 × PBS (phosphate-buffered saline), and lyophilized. The resulting lyophilized powder was subjected to XRD analysis in the same manner as in Examples 1-3.

[0182] As shown in Figure 3, the XRD analysis results confirmed that cyclosporine was amorphous, as in Examples 1-3. That is, a solid formulation consisting of a pharmaceutical composition, which is a solid dispersion in which cyclosporine is maintained in an amorphous state, was successfully prepared. The final formulation composition is shown in Table 2.

[0183] [Table 2]

[0184] <Test Example 2> A pharmacokinetic study of CyA was conducted in rats. All animals were allowed free access to food and water before administration. The solution formulation prepared in Comparative Example 1-1 and the solution formulations consisting of the pharmaceutical compositions prepared in Examples 1-1 to 1-2 and Comparative Example 1-2 were each administered intraduodenally to male Sprague-Dawley (SD) rats after undergoing duodenal cannulation surgery and at least 72 hours of recovery. In each group, 5 mg / kg of the drug was administered into the duodenum of male SD rats (6 weeks old) via cannulation. In addition, the solid formulation prepared in Comparative Example 1-3 and the solid formulations consisting of the pharmaceutical compositions prepared in Examples 1-3 to 1-5 were each dispersed in phosphate-buffered saline (PBS) and similarly administered to the duodenum of male SD rats (6 weeks old) at 5 mg / kg using cannulation.

[0185] After administration, blood samples were collected from the jugular vein over time using a heparinized syringe, and aprotinin was added as a protease inhibitor. Plasma was separated from the collected blood and measured by LC-MS / MS. The plasma concentration of CyA after administration of each formulation is shown in Figure 4.

[0186] Furthermore, the area under the plasma CyA drug concentration-time curve (0 to infinity) (AUCall) after administration of each formulation was analyzed using WinNonlin Ver. 8.3 (Pharsight), and the values ​​are shown in FIG.

[0187] As shown in Figures 4 and 5, it was revealed that the pharmaceutical composition of the hyaluronic acid derivative for oral absorption of this embodiment promotes drug absorption in the intestinal tract. Furthermore, in comparison between Example 1-1 and Comparative Example 1-2, it was confirmed that the higher the encapsulation rate of the active ingredient in the hyaluronic acid derivative, the better the drug absorption ability in the intestinal tract.

[0188] <Test Example 3> In Test Example 3, ARV-825, a compound having a site that targets E3 ligase, was formulated.

[0189] [Comparative Example 2-1] A solution formulation was prepared in the same manner as in Comparative Example 1-1, except that ARV-825 was used. The final formulation composition is shown in Table 3.

[0190] [Example 2-1] A solution formulation using a hyaluronic acid derivative (10k HA-C6-Chol-30%) was prepared. The following operations were carried out at room temperature (20°C). The freeze-dried hyaluronic acid derivative (10k HA-C6-Chol-30%) obtained in Synthesis Example 1 was dissolved at 50.0 mg / mL in a 10% sucrose aqueous solution by stirring for 12 hours. 7.2 mg of powdered ARV-825 (Medchemexpress, product number: HY-16954) was weighed into a separate vial. Next, 1.44 mL of a 50 mg / mL hyaluronic acid derivative aqueous solution was added to the vial containing the powder, followed by the addition of DMSO, PEG400, CreEL, and 10% sucrose aqueous solution in that order. The drug was complexed while stirring with a stirrer to prepare a hyaluronic acid derivative pharmaceutical composition. A solution formulation containing 10 mg / mL of hyaluronic acid derivative (10k HA-C6-Chol-30%) was prepared with a final concentration of DMSO / PEG400 / CreEL / 10% sucrose aqueous solution (5:30:2.5:62.5) (volume ratio). The final formulation composition is shown in Table 3.

[0191] [Table 3]

[0192] <Test Example 4> A pharmacokinetic study of ARV-825 was conducted in rats. The formulation prepared in Comparative Example 2-1 and the liquid formulation consisting of the pharmaceutical composition prepared in Example 2-1 were each administered intraduodenally to male SD rats after undergoing duodenal cannulation surgery and at least 72 hours of recovery. In both groups, 5 mg / kg of the formulation was administered into the duodenum of male SD rats (6 weeks old) via cannulation.

[0193] After administration, blood samples were collected from the jugular vein over time using a heparinized syringe, and aprotinin was added as a protease inhibitor. Plasma was separated from the collected blood and measured by LC-MS / MS. The plasma concentration profiles of ARV-825 and CyA (Comparative Example 1) after administration of each formulation are shown in Figure 6. In FIG. 6, Example 4-1 and Comparative Example 4-1 represent the cases where a preparation consisting of the hyaluronic acid derivative pharmaceutical composition of Example 3-1 and a solution preparation of Comparative Example 4-1 were administered, respectively.

[0194] Furthermore, the area under the plasma drug concentration-time curve (0 to infinity) (AUCall) after administration of each formulation was analyzed using WinNonlin Ver. 8.3 (Pharsight), and the values ​​are shown in FIG.

[0195] As shown in Figures 6 and 7, it has been revealed that the pharmaceutical composition of this embodiment has high intestinal absorption capacity for drugs, even for drugs with extremely low intestinal absorption capacity. This intestinal absorption capacity is thought to be due to the fact that the complex of (A) hyaluronic acid derivative and (B) active ingredient is stably maintained in the pharmaceutical composition of this embodiment, thereby maintaining supersaturation of the drug in the intestinal tract. [Industrial Applicability]

[0196] According to the present invention, it is possible to provide a pharmaceutical composition of a hyaluronic acid derivative for oral absorption that has excellent oral absorption ability of a drug.

Claims

1. (A) a hyaluronic acid derivative having a steryl group introduced therein; and (B) an active ingredient, The content of the (B) active ingredient relative to 100 parts by mass of the (A) hyaluronic acid derivative is 0.1 parts by mass or more and less than 10,000 parts by mass, The average molecular weight of the hyaluronic acid derivative (A) is 6,000 or more and 20,000 or less. A pharmaceutical composition of a hyaluronic acid derivative for oral absorption.

2. The pharmaceutical composition of claim 1, wherein the hyaluronic acid derivative (A) and the active ingredient (B) form a solid dispersion.

3. The orally absorbable hyaluronic acid derivative pharmaceutical composition according to claim 2, wherein the active ingredient (B) is amorphous.

4. 2. The pharmaceutical composition of claim 1, wherein the hyaluronic acid derivative (A) and the active ingredient (B) form nanoparticles.

5. A pharmaceutical composition of hyaluronic acid derivative for oral absorption according to any one of claims 1 to 4, wherein the content of the active ingredient (B) relative to 100 parts by mass of the hyaluronic acid derivative (A) is 10 parts by mass or more and 250 parts by mass or less.

6. The oral hyaluronic acid derivative pharmaceutical composition according to any one of claims 1 to 4, wherein the steryl group introduction rate of the hyaluronic acid derivative (A) is 5% or more but less than 50%.

7. The oral hyaluronic acid derivative pharmaceutical composition according to any one of claims 1 to 4, wherein the (B) active ingredient is at least one selected from proteins, peptides, and poorly water-soluble drugs.

8. The pharmaceutical composition of any one of claims 1 to 4, wherein the hyaluronic acid derivative (A) has one or more repeating units represented by the following general formula (I): 【Chemistry 1】 (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 -NR 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 -S-R, -NR ｂ -CO-Y ｂ It is a group selected from the group consisting of groups represented by -SR and -SSR. 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 R is a group selected from the group consisting of alkyl. ａ , R ｂ and R ｃ The alkyl portion of the 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 R is a group selected from the group consisting of alkyl. ｆ The alkyl portion of the formula may have inserted therein 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 Here, the alkylene of Y is —O—, —NR ｇ A group selected from the group consisting of - 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 R is a group selected from the group consisting of alkyl. ｇ The alkyl portion of the formula may have inserted therein a group selected from the group consisting of -O- and -NH-. Y ａ is C １－５ It is alkylene. Y ｂ is C ２－８ Alkylene or C ２－８ It is alkenylene. m is an integer of 1 or more and 100 or less.

9. The orally absorbable hyaluronic acid derivative pharmaceutical composition according to claim 8, wherein the steryl group is a cholesteryl group.

10. The pharmaceutical composition of any one of claims 1 to 4, wherein the absorption of the active ingredient (B) is promoted in at least a part of the stomach, small intestine (duodenum, jejunum, ileum), and large intestine.

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