Hyaluronic acid derivative pharmaceutical composition and pharmaceutical composition

Abstract

The present invention is a hyaluronic acid derivative pharmaceutical composition containing an active ingredient suspended in an aqueous carrier containing a hyaluronic acid derivative, the active ingredient being microparticles having a molecular weight of 100 g / mol or more and 8000 g / mol or less. The microparticles are preferably micron particles or nano particles.

Description

Technical Field

[0001] The present invention relates to a hyaluronic acid derivative pharmaceutical composition and a pharmaceutical composition. This application claims priority based on Special Application No. 2022-115840 filed in Japan on July 20, 2022, and Special Application No. 2022-143474 filed in Japan on September 9, 2022, and the contents of which are cited here. Background Art

[0002] In recent years, the number of biopharmaceuticals with proteins, peptides, and nucleic acids as active ingredients has continued to increase year by year. Biopharmaceuticals can meet the scarce medical needs that cannot be met by existing low-molecular-weight drugs.

[0003] However, biopharmaceuticals are difficult to be absorbed by the digestive tract, mucous membranes, etc., and in addition, there are problems such as instability in the body and a short half-life in the blood. Therefore, biopharmaceuticals need to be administered by frequent injections, which is a heavy burden for both patients and medical personnel. Therefore, there is a need for a drug substrate (sustained-release drug delivery system substrate) that can slowly release the active ingredient in the body after encapsulating the biopharmaceutical without losing its pharmacological activity.

[0004] Against such a background, Patent Document 1 proposes a sustained-release drug delivery system substrate composed of a hyaluronic acid derivative having excellent safety. The hyaluronic acid derivative disclosed in patent documentation 1 associates spontaneously in aqueous solution, can be in the state of keeping its biological activity, medicine, particularly biopharmaceutical efficiently.Thus, coagulate under normal saline concentration (or also can disperse under normal saline concentration), in addition retention is good in blood.This hyaluronic acid derivative, particularly when biopharmaceutical is used as active ingredient, can be used as the carrier that can be in the state of maintaining pharmacological activity, blood retention excellent blood slow-release carrier and the targeting carrier of good efficiency in enclosing a large amount of medicines, blood, also can be used as the local (such as subcutaneous etc.) slow-release carrier of persistent sustained-release medicine. Prior art literature Patent Literature

[0005] Patent Document 1: International Publication No. 2010 / 053140 Summary of the invention Problem that the invention aims to solve

[0006] The sustained-release drug matrix is ​​required to maintain the concentration of the active ingredient in the body for a long period of time and slowly release the active ingredient. In addition, it is required to suppress abnormal initial release (initial burst).

[0007] In view of the above circumstances, the present invention aims to provide a hyaluronic acid derivative pharmaceutical composition and a pharmaceutical composition which can maintain the concentration of the active ingredient in the body for a long time and reduce the abnormal initial release amount (initial burst release). Means to solve the problem

[0008] That is, the present invention includes the following embodiments. [1] A hyaluronic acid derivative pharmaceutical composition comprising an active ingredient suspended in an aqueous carrier, wherein the aqueous carrier contains a hyaluronic acid derivative, and the active ingredient is a microparticle having a molecular weight of not less than 100 g / mol and not more than 8000 g / mol. [2] According to the hyaluronic acid derivative pharmaceutical composition described in [1], the microparticles are preferably microparticles or nanoparticles. [3] The hyaluronic acid derivative pharmaceutical composition according to [1] or [2], wherein the hyaluronic acid derivative contains one or more repeating units represented by the following general formula (I). [Chemistry 1] (Where R 1 , R 2 , R 3 and R 4 Each independently consists of hydrogen atoms, C 1-6 Alkyl, formyl and C 1-6 A group selected from the group consisting of alkylcarbonyl groups. Z represents a direct bond or a peptide linker consisting of 2 to 30 arbitrary amino acid residues. X 1 For-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 A group selected from the group consisting of groups represented by -SR and -SSR. R a , R b and R c Each independently consists of hydrogen atoms, C 1-20 Alkyl, amino C 2-20 Alkyl and hydroxyl C 2-20 A group selected from the group consisting of alkyl groups.a , R b and R c The alkyl moiety can also be inserted by -O- and -NR f - A group selected from the group consisting of R f Composed of hydrogen atoms, C 1-12 Alkyl, amino C 2-12 Alkyl and hydroxyl C 2-12 A group selected from the group consisting of alkyl groups. f The alkyl portion may be interrupted by a group selected from the group consisting of -O- and -NH-. R is a hydrophobic group or a hydrophilic group. Y is C 2-30 Alkylene or -(CH2CH2O) m -CH2CH2-. The alkylene group of Y can also be inserted by -O-, -NR g -and -SS-. R g Composed of hydrogen atoms, C 1-20 Alkyl, amino C 2-20 Alkyl and hydroxyl C 2-20 A group selected from the group consisting of alkyl groups. g The alkyl portion may be interrupted by a group selected from the group consisting of -O- and -NH-. Y a C 1-5 Alkylene. Y b C 2-8 Alkylene or C 2-8 Alkenylene. m is an integer greater than or equal to 1 and less than or equal to 100.) [4] The hyaluronic acid derivative pharmaceutical composition according to [3], wherein R is a hydrophobic group. [5] The hyaluronic acid derivative pharmaceutical composition according to [4], wherein the hydrophobic group is a cholesteryl group. [6] The hyaluronic acid derivative pharmaceutical composition according to [1] or [2], wherein the hyaluronic acid derivative is precipitated or salted out in vivo or under physiological conditions. [7] The hyaluronic acid derivative pharmaceutical composition according to any one of [1] to [6], wherein the active ingredient is a poorly water-soluble drug. [8] The hyaluronic acid derivative pharmaceutical composition according to any one of [1] to [7], wherein the active ingredient is a cyclic polypeptide. [9] The hyaluronic acid derivative pharmaceutical composition according to [8], wherein the cyclic polypeptide comprises a ring size of 4 to 49 amino acids.

[10] The hyaluronic acid derivative pharmaceutical composition according to [8] or [9], wherein the cyclic polypeptide contains a non-natural amino acid.

[11] The hyaluronic acid derivative pharmaceutical composition according to any one of [8] to

[10] , wherein the cyclic polypeptide is composed of 8 or more and 20 or less amino acids.

[12] The hyaluronic acid derivative pharmaceutical composition according to any one of [1] to

[11] , wherein the administration site is subcutaneous, intramuscular, intramedullary, intraarticular, intracerebral, intravitreal, ocular surface, nose, lung, or mouth.

[13] The hyaluronic acid derivative pharmaceutical composition according to any one of [1] to

[12] , wherein the administration method is injection, eye drops or application. Effects of the Invention

[0009] According to the present invention, a hyaluronic acid derivative pharmaceutical composition and a pharmaceutical composition can be provided which can maintain the concentration of the active ingredient in the body for a long time and reduce the abnormal initial release amount (initial burst release). BRIEF DESCRIPTION OF THE DRAWINGS

[0010] [ Figure 1 ] represents the X-ray powder diffraction peak of cyclosporine. [ Figure 2 ] is a graph showing the results of changes in cyclosporine concentration in plasma when a cyclosporine suspension preparation is administered. [ Figure 3 ] is a graph showing the results of changes in cyclosporine concentration in plasma when a cyclosporine suspension preparation is administered. [ Figure 4 ] are photographs showing the results of the evaluation of precipitation in the vitreous body. [ Figure 5 ] is a graph showing the results of changes in cyclosporine concentration in plasma when a cyclosporine suspension preparation is administered. [ Figure 6 ] is a graph showing the results of changes in cyclosporine concentration in plasma when a cyclosporine suspension preparation is administered. [ Figure 7 ] is a graph showing the results of changes in cyclosporine concentration in plasma when a cyclosporine suspension preparation is administered. [ Figure 8 ] is a graph showing the results of changes in cyclosporine concentration in plasma when a cyclosporine suspension preparation is administered. [ Fig. 9 ] is a graph showing the results of changes in cyclosporine concentration in plasma when a cyclosporine suspension preparation is administered. [ Fig.10 ] is a graph showing the results of changes in cyclosporine concentration in plasma when a cyclosporine suspension preparation is administered. DETAILED DESCRIPTION

[0011] An embodiment of the present invention (hereinafter referred to as “the present embodiment”) will be described in detail below, but the present invention is not limited thereto and various modifications can be made without departing from the gist of the invention.

[0012] The following is a description of the terms used in this specification.

[0013] The “C 1-20 The term "alkyl" refers to a linear or branched alkyl group having 1 to 20 carbon atoms, such as methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, isobutyl, tert-butyl, etc. 1-4 "Alkyl" 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, etc. In C 1-20 The alkyl group also includes a C 1-12 Alkyl, C 1-6 alkyl.

[0014] The “C 1-6 The term "alkylcarbonyl" means that the alkyl portion is a C 1-6 The alkylcarbonyl of the alkyl group includes, for example, acetyl, propionyl, n-propylcarbonyl, isopropylcarbonyl, n-butylcarbonyl, sec-butylcarbonyl, isobutylcarbonyl, tert-butylcarbonyl, etc. 1-4 "Alkylcarbonyl".

[0015] The term "amino C" used in this specification 2-20 The term "alkyl" refers to a linear or branched alkyl group having 2 or more and 20 or less carbon atoms and having an amino group as a substituent. For example, the amino group may be located at the carbon atom at the terminal of the alkyl group. 2-20 The alkyl group also includes amino groups having 2 or more and 12 carbon atoms. 2-12 alkyl.

[0016] The term "hydroxyl C 2-20 The term "alkyl" refers to a linear or branched alkyl group having 2 or more and 20 or less carbon atoms and having a hydroxyl group as a substituent. For example, the hydroxyl group may be located on the carbon atom at the terminal of the alkyl group. 2-20 The alkyl group also includes a hydroxyl group having 2 or more and 12 carbon atoms. 2-12 alkyl.

[0017] The “C 2-30The term "alkylene" refers to a linear or branched divalent saturated hydrocarbon group having 2 or more and 30 carbon atoms, such as ethylene, propylene, etc., and also includes C 2-20 Alkylene, C 2-8 Alkylene, "-(CH2) n -" group (wherein n is 2 or more and 30 or less, preferably 2 or more and 20 or less, more preferably 2 or more and 15 or less).

[0018] The “C 1-5 The term "alkylene" refers to a linear or branched divalent saturated hydrocarbon group having 1 to 5 carbon atoms, and includes, for example, methylene, ethylene, and propylene.

[0019] The term "C 2-8 "Alkenylene" refers to a linear or branched divalent saturated hydrocarbon group having 2 or more and 8 carbon atoms and containing one or more double bonds, such as -CH=CH-, -C(CH3)=CH-, 2-butene-1,4-diyl, 2,4-heptadiene-1,6-diyl, 2,4,6-octatriene-1,8-diyl, etc. When geometric isomers exist, each isomer and a mixture thereof are also included.

[0020] <Hyaluronic acid derivative pharmaceutical composition> The present embodiment is a pharmaceutical composition of a hyaluronic acid derivative comprising an active ingredient suspended in an aqueous carrier containing a hyaluronic acid derivative.

[0021] <Aqueous carrier containing hyaluronic acid derivative> In the present specification, the aqueous carrier containing a hyaluronic acid derivative is an aqueous carrier containing a hyaluronic acid derivative described later. In this specification, "aqueous carrier" refers to a pharmaceutically acceptable carrier, which may include: substances approved by the Pharmaceutical and Medical Devices Administration (PMDA) of Japan and the Food and Drug Administration (FDA) of the United States as pharmaceutical excipients for injection preparations, or substances included in the Japanese Pharmacopoeia, the United States Pharmacopoeia and other well-known pharmacopoeias as pharmaceutical excipients for injection preparations for animals, more specifically for humans.

[0022] Specifically, the aqueous carrier is preferably a sterile liquid, such as water or oil, or an emulsion (such as oil / water or water / oil emulsion, etc.).

[0023] Water includes water, water for injection, physiological saline solution, phosphate buffer, phosphate buffer containing sucrose and sucrose aqueous solution, glucose aqueous solution, maltose aqueous solution, trehalose aqueous solution and glycerol aqueous solution, an aqueous solution containing polyethylene glycol, an aqueous solution containing polysorbate, an aqueous solution containing poloxamer, an aqueous solution containing cyclodextrin, an aqueous solution containing polyvinyl pyrrolidone, and an aqueous solution containing carboxymethyl cellulose.

[0024] Oils include those of petroleum, animal, vegetable, and synthetic origin, such as peanut oil, soybean oil, mineral oil, sesame oil, and the like.

[0025] In this embodiment, the aqueous carrier is preferably one or more selected from the group consisting of water, water for injection, physiological saline solution, aqueous buffer, phosphate buffer, phosphate buffer containing sucrose, sucrose aqueous solution, glucose aqueous solution and glycerol aqueous solution. Among them, the aqueous carrier is more preferably water, phosphate buffer, phosphate buffer containing sucrose or sucrose aqueous solution.

[0026] The aqueous carrier may contain diluents, supplements, excipients, stabilizers, isotonic agents, buffers, pH adjusters, preservatives, bactericides or antimicrobial agents, thickeners, etc., and solvents for administration together with the compound.

[0027] In the present specification, "containing an active ingredient suspended in an aqueous carrier containing a hyaluronic acid derivative" means that the active ingredient particles are not dissolved in the aqueous carrier containing a hyaluronic acid derivative, but are dispersed and floated in the aqueous carrier containing a hyaluronic acid derivative. In this specification, the term "suspended" means suspended in a broad sense, and means that a dispersoid is dispersed in a dispersion medium. One embodiment of a suspension is a dispersion of solid particles in a liquid. In this specification, the dispersed substance can be solid, liquid or gas, and the dispersion medium can be liquid. In addition, liquid-liquid phase separation in which the dispersion medium and the liquid dispersed substance are dispersed immiscibly also belongs to an embodiment of suspension.

[0028] The active ingredient particles may be uniformly suspended in the aqueous carrier or non-uniformly suspended in the aqueous carrier, but are preferably uniformly suspended.

[0029] Whether the active ingredient particles are suspended in an aqueous carrier containing a hyaluronic acid derivative depends not only on the upper limit of the solubility of the aqueous carrier, but also on the particle size of the particles. Because the particles are relatively large, all the particles precipitate instantly (for example, within 1 second) and do not float in the aqueous carrier, which does not belong to the situation of "suspending in an aqueous carrier". From the viewpoint of more stably "suspending in an aqueous carrier", the particle size of the particles is preferably in the range of 1nm to 1000μm, more preferably 5nm to 100μm, further preferably 10nm to 10μm, further preferably 50nm to 10μm, further preferably 50nm to 5μm, further preferably 50nm to 1μm, and most preferably 50nm to 500nm.

[0030] In addition, the period during which the active ingredient is "suspended in the aqueous carrier" is preferably more than one year from the perspective of the drug. Even if sedimentation occurs once, if it can be uniformly dispersed again to a state of "suspended in the aqueous carrier" by stirring before administration, it can also be determined to be "containing an active ingredient suspended in an aqueous carrier containing a hyaluronic acid derivative."

[0031] For example, the active ingredient can be determined to be solid by visual observation or microscopic observation. Unlike liquids and gases, solids can maintain a certain shape. In addition, solids can also be determined by the difficulty in deformation.

[0032] In the present embodiment, whether the active ingredient is suspended in the aqueous carrier containing the hyaluronic acid derivative can be determined by the turbidity test method of the general test method recorded in the Japanese Pharmacopoeia. As the turbidity test method, it can be judged by visual method or photoelectric photometry such as transmitted light determination method, scattered light determination method, and transmission scattering method. When it is a medicine in a suspended state, it is recorded in the drug manual, for example. It is not particularly limited, such as being recorded as a suspension, a suspension injection, an aqueous suspension injection, and a suspension eye drop.

[0033] It is more preferred that the active ingredient particles do not settle in the aqueous carrier.

[0034] When the microparticles can maintain a dispersed state and suspend in an aqueous carrier, when administered into a living body, the hyaluronic acid derivative gels or precipitates at a physiological saline concentration, and the microparticles can be retained and carried in the hyaluronic acid derivative gel or precipitate, thereby not diffusing from the administration site and being stably retained. As a result, the amount and surface area of ​​the active ingredient can be locally maintained, a stable long-term sustained release is exerted, and abnormal initial release amount is reduced.

[0035] The active ingredient particles are uniformly maintained in the aqueous carrier without settling, and preferably precipitate together with the hyaluronic acid derivative when the hyaluronic acid derivative is gelled or precipitated at a physiological saline concentration.

[0036] For example, when only active ingredient microparticles are administered in vivo without using hyaluronic acid derivatives, the microparticles diffuse in vivo after administration, and the initial release amount is easily abnormal, and the long-term sustained release function can not be brought into play. In addition, the microparticles can not maintain a dispersed state, and coagulation occurs between the microparticles and the surface area decreases. As a result, the dissolution rate of the active ingredient decreases, and sufficient blood concentration can not be reached, so the expected drug effect can not be obtained. In addition, when not using hyaluronic acid derivatives for administration, the microparticles are not retained in the administration site, so the existence state of the microparticles of each administration is different, which can cause differences.

[0037] As a conventional technique, a method for forming a complex of a hyaluronic acid derivative and an active ingredient is known. “Forming a complex of a hyaluronic acid derivative and an active ingredient” means holding the active ingredient in a physically cross-linked region based on the hydrophobic interaction of the hyaluronic acid derivative.

[0038] When forming a complex of a hyaluronic acid derivative and an active ingredient, the amount of the active ingredient adsorbed is limited to 1 part by mass to a maximum of 100 parts by mass relative to 100 parts by mass of the hyaluronic acid derivative. On the other hand, the hyaluronic acid derivative pharmaceutical composition of the present invention is not particularly limited as long as the active ingredient is suspended in an aqueous carrier.

[0039] When forming the complex of hyaluronic acid derivative and active ingredient, active ingredient is solubilized into liquid state, therefore even if hyaluronic acid derivative gelates or precipitates after being administered in vivo, it will be released earlier and transferred to blood. In addition, due to the interaction of active ingredient, when being administered in vivo, the precipitation of hyaluronic acid derivative decreases, for example, when the hyaluronic acid derivative in the pharmaceutical composition is not completely precipitated 5%, the active ingredient forming the complex with the hyaluronic acid derivative is rapidly transferred to blood, i.e., becomes the main cause of initial burst release. On the other hand, the active ingredient of the hyaluronic acid derivative pharmaceutical composition of the present invention is solid state, not only can not damage the precipitation performance of hyaluronic acid derivative when being administered in vivo, and even when the hyaluronic acid derivative in the pharmaceutical composition is not completely precipitated 5%, the active ingredient slowly dissolves and absorbs because of solubility rate-limiting, and initial burst release can be suppressed to the greatest extent.

[0040] <Hyaluronic acid derivatives> The hyaluronic acid derivative used in the present embodiment is preferably a hyaluronic acid derivative having one or more repeating units represented by the following general formula (I).

[0041] [Chemistry 2]

[0042] (Where R 1, R 2 , R 3 and R 4 Each independently consists of hydrogen atoms, C 1-6 Alkyl, formyl and C 1-6 A group selected from the group consisting of alkylcarbonyl. Z represents a direct bond or a peptide linker composed of 2 or more and 30 or less arbitrary amino acid residues. X 1 For-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 A group selected from the group consisting of groups represented by -SR and -SSR. R a , R b and R c Each independently consists of hydrogen atoms, C 1-20 Alkyl, amino C 2-20 Alkyl and hydroxyl C 2-20 A group selected from the group consisting of alkyl. a , R b and R c The alkyl moiety can also be inserted by -O- and -NR f - A group selected from the group consisting of R f Composed of hydrogen atoms, C 1-12 Alkyl, amino C 2-12 Alkyl and hydroxyl C 2-12 A group selected from the group consisting of alkyl groups. f The alkyl portion may be interrupted by a group selected from the group consisting of -O- and -NH-. R is a hydrophobic group or a hydrophilic group. Y is C 2-30 Alkylene or -(CH2CH2O) m -CH2CH2-. The alkylene group of Y can also be inserted by -O-, -NR g -and -SS-. R g Composed of hydrogen atoms, C 1-20 Alkyl, amino C 2-20 Alkyl and hydroxyl C2-20 A group selected from the group consisting of alkyl groups. g The alkyl portion may be interrupted by a group selected from the group consisting of -O- and -NH-. Y a C 1-5 Alkylene. Y b C 2-8 Alkylene or C 2-8 Alkenylene. m is an integer greater than or equal to 1 and less than or equal to 100.)

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

[0044] [Chemistry 3]

[0045] (Where R 1 , R 2 , R 3 and R 4 Each independently consists of hydrogen atoms, C 1-6 Alkyl, formyl and C 1-6 A group selected from the group consisting of alkylcarbonyl. X is -NR a -Y-NR b -COO-R represents a hydrophobic group. a and R b Each independently consists of hydrogen atoms and C 1-6 A group selected from the group consisting of alkyl. R is a hydrophobic group or a hydrophilic group. Y is C 2-30 Alkylene or -(CH2CH2O) m -CH2CH2-, m is an integer of 1 to 100. )

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

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

[0048] [Repeating unit (I)] The group "-ZN(R a)YX 1 " includes the following 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 (wherein mz is an integer of 2 to 30, R 8 is a hydrogen atom or a methyl group, and R and m are as defined in the present specification. A group selected from the group consisting of the groups represented by ).

[0049] As this group, -NH-(CH2) is preferred. mz -NH-COO-R; -NH-(CH2CH2O) m -CH2CH2-NH-COO-R and -NH-(CH2) mz -SSR (wherein mz, ​​R and m are as defined in the present specification).

[0050] (Z) In the general formula (I), Z is preferably directly bonded (directly bonded). In addition, in other embodiments, when Z is a peptide linker, X 1 Preferably -NRb -COO-R. Further, in other embodiments, Z may also be -NH-[CH(-Z a )-CONH] n-1 -CH(-Z a )-CO-, wherein n is an integer of 2 to 30, and Z a Each independently represents H2N-CH(-Z a )-COOH in an α-amino acid. The peptide linker forms a bond with the carboxyl group of the glucuronic acid moiety at the N-terminus and with -N(-R a )-YX 1 The amino acids that can be used as amino acid residues of the peptide linker include, for example, 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, valine and other natural (L-type) amino acids and their dextrorotatory (D-type) amino acids, and all α-amino acids including synthetic amino acids can be used. That is, as Z a , for example, -CH3, H2NC(NH)NH(CH2)3-, H2NCOCH2-, etc. In addition, n Zs may be the same or different. n is an integer of 2 to 30, preferably 2 to 10, more preferably 2 to 4. Preferred examples of peptide linkers include -Gly-Phe-Leu-Gly-, -Asn-Phe-Phe-, -Phe-Phe-, Phe-Gly-, etc.

[0051] Wherein the so-called "joint" can use any peptide joint or synthetic compound joint that can be introduced by genetic engineering, but in the hyaluronic acid derivative (A) of the present embodiment, it is preferably a peptide joint. The length of the peptide joint is not particularly limited, and those skilled in the art can appropriately select it according to the purpose, and the preferred length is more than 2 amino acids (the upper limit is not particularly limited, usually less than 30 amino acids, preferably less than 20 amino acids), and 15 amino acids are particularly preferred. The peptide joints contained in the (A) hyaluronic acid derivative can all use peptide joints of the same length, or peptide joints of different lengths can be used.

[0052] (Y) In the general formula (I), Y is preferably -(CH2) n1 -and-(CH2CH2O) m1-CH2CH2- (wherein n1 is an integer from 2 to 20, preferably an integer from 2 to 15, more preferably an integer from 2 to 12, and further preferably an integer from 2 to 6. m1 is an integer from 1 to 4). Specifically, -(CH2)2-, -(CH2)6-, -(CH2)8-, -(CH2) 12 In addition, from the viewpoint of achieving high solubility in pure water or at low salt concentrations and showing high precipitation forming ability at physiological saline concentrations, Y is preferably -(CH2)2-, -(CH2)6-, -(CH2)8- and -(CH2) 12 -, more preferably -(CH2)6-.

[0053] Y can 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-, or -(CH2CH2O)2-CH2CH2-SS-(CH2CH2O)2-CH2CH2-.

[0054] (Y a ) As Y a , preferably -CH2- or -CH2-CH2-.

[0055] (Y b ) As Y b , preferably -CH2-CH2-, -CH(CH3)CH2-, 2-butene-1,4-diyl, 2,4-heptadiene-1,6-diyl or 2,4,6-octatriene-1,8-diyl, more preferably -CH2-CH2- or -CH(CH3)CH2-.

[0056] As the group "-ZN(R a )YX 1Specific examples of "" include: -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-NH-(CH2)3-NH-COO-cholesteryl, -NH-(CH2)4-N(-(CH2)3-NH2)-COO-cholesteryl, -NH-(CH2)3-NH-(CH2)4-N(-(CH2)3 -NH2)-COO-cholesteryl, -NH-(CH2)3-NH-(CH2)4-N(-(CH2)3-NH-(CH2)3-NH2)-COO-cholesteryl, -NH-(CH2)3-NH-(CH2)4-N(-(CH2)3-NH2)-CO-NH-cholesteryl, -NH-(CH2)3-NH-(CH2)4-N(-(CH2)3-NH2)-CO-cholesteryl, -NH-(CH2)3-NH-(CH2)4-N(-(CH2)3-NH2)-CO-cholesteryl, -NH-(CH2)3-NH-(CH2)4-N(-(CH2)3-NH2)-cholesteryl, etc. Preferred groups include "-ZN(R a )YX 1 ”, R a , R b and R c is a hydrogen atom, and Y is a straight-chain C 2-30 Alkylene or -(CH2CH2O) m -CH2CH2-,Y a A straight chain C 1-5 Alkylene or Y b A straight chain C 2-8 Alkylene or straight chain C 2-8 Alkenylene.

[0057] (R) R is a hydrophilic group or a hydrophobic group, preferably a hydrophobic group. As the hydrophilic group, there can be mentioned: derivatives with a polyethylene glycol skeleton, derivatives with a polyethyleneimine skeleton, derivatives with a polylysine skeleton, derivatives with a polyallylamine skeleton, derivatives with a natural amino acid skeleton, derivatives with a non-natural amino acid skeleton, and derivatives with a polydimethylsiloxane skeleton.

[0058] As the hydrophobic group, a steryl group is preferred.

[0059] The term "steryl" used in this specification sheets is not particularly limited as long as it is a group with a steroid skeleton. Wherein as steroid, specifically include: cholesterol, cholestanol, campesterol, ergosterol, stigmasterol, coprostanol, stigmasterol, sitosterol, lanosterol, ergosterol, sagostanol, bile acid, testosterone, estradiol, progesterone, cortisol, cortisone, aldosterone, corticosterone, deoxycorticosterone etc. As steryl, cholesteryl, stigmasteryl, lanosteryl, ergosteryl etc. can be included, wherein preferably cholesteryl (especially cholesteryl-5-ene-3β-base).

[0060] [Repeating unit (Ia)] In the general formula (Ia), X is preferably -NH-(CH2)2-NH-COO-cholesteryl, -NH-(CH2)6-NH-COO-cholesteryl, -NH-(CH2) 12 -NH-COO-cholesterol or -NH-(CH2CH2O)2-CH2CH2-NH-COO-cholesterol, more preferably -NH-(CH2)2-NH-COO-cholesterol, -NH-(CH2)6-NH-COO-cholesterol or -NH-(CH2CH2O)2-CH2CH2-NH-COO-cholesterol.

[0061] The hyaluronic acid derivative may further contain a repeating unit represented by the general formula (II) in addition to the repeating unit (I) (hereinafter, sometimes referred to as "repeating unit (II)").

[0062] [Chemistry 4]

[0063] (Where R 1a , R 2a , R 3a and R 4a Each independently consists of hydrogen atoms, C 1-6 Alkyl, formyl and C 1-6 A group selected from the group consisting of alkylcarbonyl. a Composed of hydroxyl and -OQ + The group selected from the group consisting of + is the counter cation.)

[0064] When the hyaluronic acid derivative contains two or more repeating units (II), the repeating units may be the same or different. In other embodiments, the hyaluronic acid derivative may be a hyaluronic acid derivative consisting essentially of the repeating unit (I), the repeating unit (Ia) and the repeating unit (II).

[0065] [Repeating unit (II)] In the general formula (II), as long as Q + There is no particular limitation on the counter cation that can form a salt with a carboxyl group in water. When the counter cation is divalent or higher, it forms a salt with multiple carboxyl groups according to the valence. Examples of counter cations include metal ions such as lithium ions, sodium ions, rubidium ions, cesium ions, magnesium ions, and calcium ions; Formula: N + R j R k R l R m (Where R j , R k , R l and R m are independently selected from hydrogen atoms and C 1-6 alkyl group) represented by ammonium ions, etc. + It is preferably a sodium ion, a potassium ion or a tetraalkylammonium ion (for example, a tetra-n-butylammonium ion). j , R k , R l and R m Preferably from C 1-6 The same group selected from the group consisting of alkyl groups is preferably n-butyl.

[0066] R 1 , R 2 , R 3 and R 4 and R 1a , R 2a , R 3a and R 4a Preferably, all of them are hydrogen atoms. a and R b Both are preferably hydrogen atoms.

[0067] Among them, the hyaluronic acid derivative is preferably a hyaluronic acid derivative substantially composed of repeating units (I) and repeating units (II). For the hyaluronic acid derivative, among the repeating units of the disaccharide composed of D-glucuronic acid and N-acetyl-D-glucosamine contained in the derivative, for example, more than 80%, preferably more than 90%, and more preferably more than 95% are repeating units (I) and repeating units (II). The hyaluronic acid derivative can be composed only of repeating units (I) and repeating units (II).

[0068] In the hyaluronic acid derivative of the present embodiment, the introduction rate of the steryl group relative to the repeating unit of the disaccharide derived from the hyaluronic acid derivative (hereinafter referred to as "steryl group introduction rate") is preferably 0.1% or more and less than 50%, more preferably 5% or more and less than 45%, further preferably 10% or more and less than 40%, and particularly preferably 15% or more and less than 35%. By introducing the steryl group within the above range, the formulation stability of the hyaluronic acid derivative pharmaceutical composition containing the active ingredient particles in the pharmaceutical composition is improved, and aggregation and precipitation occur at the concentration of physiological saline, so that the drug can be released slowly.

[0069] Available through 1 The steryl introduction rate can be measured by H-NMR measurement. 1 The integral value of the peak derived from the steryl group of the hyaluronic acid derivative in the H-NMR spectrum and the integral value of the peak derived from the acetyl group of N-acetyl-D-glucosamine contained in the hyaluronic acid derivative (COCH3, 1.6 ppm or more and 2.0 ppm or less, 3H) are calculated based on the following formula. H Specifically, it can be measured, for example, by the method described in the Examples described below.

[0070] [Steryl introduction rate] (%) = [(integral value of the peak derived from steryl group × 3 / n H ) / (peak integration value of acetyl group derived from N-acetyl-D-glucosamine)]×100

[0071] The weight average molecular weight of the hyaluronic acid derivative is not particularly limited, but from the viewpoint of increasing the number of steryl groups introduced per molecule of the hyaluronic acid derivative to form a complex with the active ingredient, and from the viewpoint of increasing molecular entanglement and improving blood retention, hyaluronic acid derivatives with a larger molecular weight are preferred.

[0072] The weight average molecular weight of such a hyaluronic acid derivative is preferably 4000 (4k) to 1,000,000 (1,000k), more preferably 5k to 500k, further preferably 7k to 300k, particularly preferably 7k to 100k.

[0073] Since the weight average molecular weight of the hyaluronic acid derivative is above the above lower limit, it is possible to further improve molecular entanglement and further improve retention in the blood. On the other hand, since the weight average molecular weight of the hyaluronic acid derivative is above the above lower limit, it is possible to suppress the rise in viscosity and dissolve a higher concentration of the hyaluronic acid derivative in a pharmaceutical composition. The weight average molecular weight of the hyaluronic acid derivative can usually be adjusted by using a raw material with a corresponding molecular weight.

[0074] In more detail, from the viewpoint of viscosity and dispersibility, the weight average molecular weight of the hyaluronic acid derivative is preferably 100k or less, further preferably 50k or less, particularly preferably 20k or less, and most preferably 15k or less. The unit of molecular weight is Da. From the viewpoint of manufacturing the hyaluronic acid derivative, it is preferably 4k to 20k, particularly preferably 6k to 16k, and most preferably 8k to 12k.

[0075] From the viewpoint that the hyaluronic acid derivative strongly exhibits the properties of hyaluronic acid, for example, in order to be strongly recognized by the CD44 receptor, the molecular weight is preferably 100 k or more, particularly preferably 200 k or more, and most preferably 300 k or more.

[0076] From the viewpoint of controlling the viscosity and precipitation rate of the hyaluronic acid derivative, it is preferably 8 k or more, particularly preferably 20 k or more, and most preferably 30 k or more.

[0077] Herein, the "molecular weight of the hyaluronic acid derivative" is the weight average molecular weight determined by size exclusion chromatography with multi-angle light scattering detector (SEC-MALS).

[0078] [Method for producing hyaluronic acid derivatives] Hyaluronic acid derivatives, for example, can be manufactured by the method described in International Publication No. 2010 / 053140 if the R is a hydrophilic group. When the R is a hydrophilic group, an amino group is introduced into the terminal of the derivative having a hydrophilic group by any method, and then a condensing agent such as DMT-MM (4-(4,6-dimethoxy-1,3,5-triazine-2-yl)-4-methylmorpholine hydrochloride) is used to manufacture the hyaluronic acid derivative by condensation reaction with the carboxyl group (-COOH or -COONa) of hyaluronic acid. In addition, when the terminal of the compound to be introduced has a hydroxyl group (-OH), any condensing agent can be used to produce it by dehydration condensation. In addition, the hydroxyl group of hyaluronic acid and the functional group of the introduced group can also be reacted to form a chemical bond. As the introduction method other than the condensing agent, it can be enumerated: enzyme reaction, dehydration condensation based on heating, the method using a metal catalyst, etc.

[0079] In the present specification, the hyaluronic acid derivative is preferably precipitated or salted out in vivo or under physiological conditions. Whether or not a hyaluronic acid derivative is precipitated or salted out in vivo or under physiological conditions can be confirmed by the following method. First, a hyaluronic acid derivative dissolved to a desired concentration is added to a phosphate buffer or PBS containing BSA. Then, the mixture is incubated at 37°C and centrifuged. If solid-liquid separation can be performed, it is determined that the hyaluronic acid derivative can be precipitated or salted out in the physiological saline. The incubation time can be set arbitrarily.

[0080] The concentration of the phosphate buffer used is, for example, a final concentration of 1×PBS or PBS containing 0.8% BSA. The centrifugal conditions during centrifugal separation can be changed appropriately, and one example is 9800 g.

[0081] In one embodiment of the present invention, 150 μL of 4×PBS is added to 450 μL of a hyaluronic acid derivative dissolved to a desired concentration, and the mixture is incubated under the above conditions. The supernatant of the hyaluronic acid derivative precipitated by centrifugation is further quantified using size exclusion chromatography (SEC), thereby determining the proportion of the hyaluronic acid derivative precipitated or salted out under physiological conditions.

[0082] In one embodiment of the present invention, the hyaluronic acid derivative can be confirmed to precipitate or salt out under physiological saline conditions by actually administering the compound to a living body or administering the compound to a collected tissue to confirm precipitation.

[0083] The proportion of the precipitate is preferably 10 to 100% of the total amount of the dissolved hyaluronic acid derivative, more preferably 40% or more, 50% or more, 60% or more, more preferably 70% or more, further preferably 80% or more, and most preferably 90% or more.

[0084] When using hyaluronic acid derivatives that precipitate or salt out in vivo or under physiological conditions, since the aggregation or diffusion of active ingredient particles contained in the pharmaceutical composition can be inhibited by gelation after administration into the body, it is relatively easy to provide a hyaluronic acid derivative pharmaceutical composition that can maintain the concentration of the active ingredient in the body for a long time without deviation and reduce the abnormal initial release amount (initial burst release).

[0085] <Active Ingredients> The active ingredient is not particularly limited as long as it is fine particles having a molecular weight of 100 g / mol or more and 8000 g / mol or less. The molecular weight of the microparticles is preferably from 200 g / mol to 8000 g / mol, more preferably from 200 g / mol to 5000 g / mol, further preferably from 300 g / mol to 4000 g / mol, further preferably from 500 g / mol to 4000 g / mol, particularly preferably from 700 g / mol to 3000 g / mol, further particularly preferably from 900 g / mol to 3000 g / mol, particularly preferably from 1000 g / mol to 3000 g / mol, and most preferably from 1000 g / mol to 2000 g / mol.

[0086] When the molecular weight of the microparticles is within the above range, the microparticles can be formed into nanoparticles or microparticles, and can float freely in an aqueous carrier without sedimentation.

[0087] In this specification, the molecular weight of active ingredient microparticles is a value calculated from the molecular formula of the compound.

[0088] In this embodiment, the microparticles are preferably microparticles or nanoparticles. Micronized particles are particles having a particle size of 1 μm or more and less than 1000 μm as measured by laser diffraction. Nanoparticles refer to particles having a particle size of 1 nm or more and less than 1000 nm as measured by dynamic light scattering (DLS).

[0089] As the active ingredient that can be used in this embodiment, there is no particular limitation as long as it is the above-mentioned microparticles, and examples thereof include: low molecular weight compounds, medium molecular weight compounds, pharmaceutically active peptides or proteins, and nucleic acids. Among them, low molecular weight compounds, medium molecular weight compounds, pharmaceutically active peptides or proteins are more preferred. Low molecular weight compounds, medium molecular weight compounds, and pharmaceutically active peptides are even more preferred. The medium molecular weight compound can be a cyclic polypeptide.

[0090] The microparticles may be crystalline or non-crystalline (amorphous).

[0091] The applicable diseases of the pharmaceutical composition of the present embodiment are not particularly limited, and can be widely used for preventing and treating diseases known at present or diseases to be discovered in the future. It can be a chronic disease or an acute disease. Examples of diseases known at present include: cancer, infectious diseases, immune diseases, inflammatory relaxation, allergic diseases, skin diseases, hypertension, diabetes, neurological diseases, hereditary diseases, cardiovascular diseases, cerebrovascular diseases, respiratory diseases, eye diseases, ear diseases, bone and joint diseases, pain, etc.

[0092] When the active ingredient in the present invention is a poorly water-soluble drug, the present invention can be used more effectively. A poorly water-soluble drug refers to a drug classified as slightly soluble, slightly poorly soluble, poorly soluble, extremely poorly soluble, or almost insoluble among drugs defined as extremely soluble, readily soluble, slightly soluble, slightly poorly soluble, poorly soluble, extremely poorly soluble, or almost insoluble in the terminology indicating solubility in the 17th revised Japanese Pharmacopoeia.

[0093] Specifically, the active ingredient may be a poorly water-soluble drug having a solubility in water of 1 mg / mL or less. Even if the pharmaceutical composition of the present invention is such a poorly water-soluble drug, it can be administered at a high dose without using an organic solvent, reducing the amount of highly toxic surfactant used, or without using a highly toxic surfactant at all.

[0094] In this embodiment, the poorly water-soluble drug may be a poorly water-soluble peptide. The poorly water-soluble peptide may be a peptide having acidic amino acids, basic amino acids, or neutral amino acids. The pH of the aqueous carrier may be appropriately selected in consideration of the isoelectric point.

[0095] In this embodiment, the active ingredient is preferably a cyclic polypeptide. The size of the ring is preferably a cyclic polypeptide consisting of 4 to 49 amino acids, more preferably 6 to 30, and most preferably 8 to 25 amino acids. The cyclic polypeptide may also contain natural amino acids or non-natural amino acids, among which non-natural amino acids are preferred. The number of amino acids constituting the cyclic polypeptide is preferably 8 or more and 20 or less.

[0096] [Low molecular weight compounds] Low molecular weight compounds refer to compounds with a molecular weight of less than about 500, and examples thereof include: anticancer agents (e.g., alkylating agents, antimetabolites, alkaloids, etc.), immunosuppressants, anti-inflammatory agents (steroids, non-steroidal anti-inflammatory agents, etc.), anti-rheumatic agents, antibacterial agents (β-lactam antibiotics, aminoglycoside antibiotics, macrolide antibiotics, tetracycline antibiotics, new quinolone antibiotics, sulfonamides, etc.), etc.

[0097] In addition, as an active ingredient, when the above-mentioned hyaluronic acid derivative has a steryl group, it is preferred to use an active ingredient with high hydrophobicity, i.e., poor water solubility, from the perspective of being able to fully exert the interaction with the steryl group. In addition, "poor water solubility" means that the amount of water required to dissolve 1g of solute in the 17th revision of the Japanese Pharmacopoeia is 30mL or more.

[0098] Examples of the poorly water-soluble solid active ingredient include: antipyretic analgesics such as rebamipide, ibuprofen, benzoic acid, ethsalamide, caffeine, camphor, quinine, calcium gluconate, dimethylhexanol, sulfonamide, theophylline, theobromine, riboflavin, toluenepropanol, phenobarbital, aminophylline, thiosemicarbazide, quercetin, rutin, salicylic acid, theophylline sodium salt, pyrabital, quinine hydrochloride, irgapyrin, digoxin, griseofulvin, phenacetin, etc., nervous system drugs, sedatives and hypnotics, muscle relaxants, hypotensive agents, antihistamines, etc.; acetylspiramycin, ampicillin, erythromycin, guitaramycin Antibiotics such as chloramphenicol, triacetyloleandomycin, nystatin, and colistin sulfate; steroid hormones such as methyltestosterone, methylandrostenediol, progesterone, estradiol benzoate, ethinylestradiol, deoxycorticosterone acetate, cortisone acetate, hydrocortisone, hydrocortisone acetate, and prednisolone; non-steroidal yolk hormones such as dienystrol, hexestrol, diethylstilbestrol, diethylstilbestrol dipropionate, and chlorethoxyquin; other fat-soluble vitamins and other pharmaceutical active ingredients listed in the Japanese Pharmacopoeia, the Standards for Drugs Outside the Japanese Pharmacopoeia, the United States Pharmacopoeia (USP), the United States National Formulary (NF), and the European Pharmacopoeia (EF). One of these active ingredients may be used, or two or more may be used in combination.

[0099] The active ingredient can also be a water-insoluble oily or liquid component. As a water-insoluble oily or liquid active ingredient, for example, teprenone, indomethacin farnesate, menatetrenone, phytomenadione, vitamin A oil, amyl alcohol, vitamin D, vitamin E and other vitamins, DHA (docosahexaenoic acid), EPA (eicosapentaenoic acid), liver oil and other higher unsaturated fatty acids, coenzyme Q, orange oil, lemon oil, peppermint oil and other oil-soluble flavorings such as "Japanese Pharmacopoeia", "Japanese Pharmacopoeia Extra-Pharmacopoeia Standards", "United States Pharmacopoeia (USP)", "National Formulary of the United States (NF)", "European Pharmacopoeia (EP)" and other pharmaceutical active ingredients recorded. Although there are various homologues and derivatives in vitamin E, as long as it is liquid at room temperature, there is no particular limitation, for example, dl-α-tocopherol, acetic acid dl-α-tocopherol, d-α-tocopherol, acetic acid d-α-tocopherol, etc. can be mentioned. One type selected from these active ingredients may be used alone, or two or more types may be used in combination.

[0100] The active ingredient may also be a semi-solid active ingredient that is poorly soluble in water. Examples of the semi-solid active ingredient that is poorly soluble in water include: Chinese herbal medicines or crude drug extracts such as earthworm, licorice, cinnamon bark, peony root, peony bark, valerian, sansho, dried ginger, tangerine peel, ephedra, nandina domestica, cherry peel, polygala, platycodon, plantago, plantain, lycoris, polygala, Fritillaria, fennel, phellodendron, coptis root, zedoaria, chamomile, gentian, bezoar, bear gall, adenophora, ginger, atractylodes, clove, tangerine peel, atractylodes, bamboo ginseng, carrot, kudzu root soup, cassia twig soup, fragrant susan, purple hu guizhi soup, small purple hu soup, small green dragon soup, ophiopogon soup, pinellia and magnolia bark soup, and ephedra soup; oyster meat extract, propolis and propolis extract, coenzyme Q, etc. One selected from these active ingredients may be used, or two or more may be used in combination.

[0101] [Medium molecular weight compounds] In this specification, "medium molecule" is neither a low molecule (an organic compound with a molecular weight of about 500) nor a high molecule (a protein with a molecular weight of 10,000 or more), but refers to a peptide, a macrolide compound, and a natural product or their derivatives with a molecular weight of about 500 to 5,000. Preferably, it is a peptide with a molecular weight of about 500 to 2,000, i.e., a linear or cyclic polypeptide with about 5 to 20 amino acid residues. As a peptide, a cyclic polypeptide is preferred, and the details thereof are described below. Macrolide compounds are a general term for compounds that are macrolides and have 12 or more rings. For example, FK506, rapamycin, etc. are mentioned.

[0102] [Nucleic Acids] Examples of nucleic acids include DNA and RNA, and include short-chain interfering RNA (siRNA), double-stranded RNA (dsRNA), microRNA (miRNA), short-chain hairpin RNA (shRNA), and aptamers.

[0103] The active ingredient may be a Rho kinase inhibitor, an endothelin A receptor inhibitor, a transmembrane conductance regulator (CFTR) regulator, a TRPV1 inhibitor, an NK1 receptor inhibitor, a purine receptor inhibitor, an angiotensin receptor inhibitor, a peroxisome proliferator-responsive receptor, a P2Y receptor inhibitor, a VEGF inhibitor, or the like, or may be an active ingredient having two of these inhibitory effects at the same time.

[0104] The content of the active ingredient in the pharmaceutical composition of the present embodiment depends on the structure of the active ingredient, and can be 0.001 mass parts or more and 10,000 mass parts or less, preferably 0.1 mass parts or more and 8000 mass parts or less, more preferably 1.0 mass parts or more and 6000.0 mass parts or less, further preferably 15 mass parts or more and 4000.0 mass parts or less, particularly preferably 30 mass parts or more and 2000 mass parts or less, and most preferably 100 mass parts or more and 1000 mass parts or less, relative to 100 mass parts of the hyaluronic acid derivative.

[0105] Alternatively, in the pharmaceutical composition of the present embodiment, the content of the active ingredient is not particularly limited. For example, relative to 100 parts by mass of the pharmaceutical composition, it is preferably 0.0001 parts by mass or more and 30.0 parts by mass or less, more preferably 0.001 parts by mass or more and 10 parts by mass or less, and further preferably 0.01 parts by mass or more and 3 parts by mass or less.

[0106] <Other additives> The pharmaceutical composition of the present embodiment can be administered individually, or can also be administered in a common manner together with a pharmacologically acceptable carrier. When used in conjunction with a pharmacologically acceptable carrier, for example, the above-mentioned hyaluronic acid derivative and the above-mentioned active ingredient can also be mixed, and adjuvants and water as required or physiologically acceptable liquids (such as normal saline, aqueous ethanol, phosphate buffered saline (PBS)) etc. in addition thereto, physiologically acceptable buffer, excipient, carrier (vehicle), preservative, stabilizer, adhesive, freeze-drying aid etc. can also be contained.

[0107] The aqueous carrier of the pharmaceutical composition of this embodiment may contain one or more surfactants. Among them, nonionic surfactants are preferred, and nonionic surfactants suitable for use as preparations described in this specification can be selected from, for example, polysorbates, poloxamers, polyoxyethylene castor oil derivatives (e.g., Cremophor EL, Cremophor RH60), bile salts, lecithin, 12-hydroxystearic acid-polyethylene glycol copolymers (e.g., Solutol HS15), tyloxapol, etc.

[0108] The pharmaceutical composition of this embodiment may include one or more water-soluble polymers. A water-soluble polymer is a pharmaceutically acceptable polymer that can be dissolved or dispersed in water. For example, a water-soluble polymer may also have the function of strengthening the viscosity of a suspension (suspension) and stabilizing the effective ingredients of microparticles or nanoparticles to counteract the potential harmful effects of particle aggregation or other formulation components. The water-soluble polymer applicable in this embodiment may be selected from plant gums, such as alginate, pectin, guar gum and xanthan gum, chemical starch, polyvinyl pyrrolidone (PVP), hydroxypropyl methylcellulose (HPMC), methylcellulose and other cellulose derivatives, such as sodium carboxymethylcellulose, hydroxypropyl cellulose, etc. The pharmaceutical composition of this embodiment may contain poloxamers such as poloxamer 188 as a water-soluble polymer. Poloxamer 188 is both a polymer and a surfactant. In other embodiments, the pharmaceutical composition of this embodiment may contain povidone K17 as a water-soluble polymer.

[0109] The pharmaceutical composition of this embodiment may contain one or more cellulose-based polymers. According to patent WO2022210784A1, it is reported that an aqueous suspension with improved redispersibility can be provided. By further mixing zinc salts or silver salts, an aqueous suspension with further improved dispersibility can be provided, and an appropriate selection can be made from the combination considering the compatibility with drugs or other additives. "Zinc salt" refers to a suspension containing zinc ions (Zn 2+ ) as a salt (compound of zinc and organic acid or inorganic acid) or zinc oxide. For example, zinc halides such as zinc chloride, zinc bromide or zinc fluoride, zinc sulfate, zinc acetate, zinc phosphate, zinc carbonate, zinc hydroxide, zinc citrate, zinc lactate, lead gluconate, zinc oxide or hydrates thereof. "Silver salt" refers to a compound containing silver ions (Ag + ) as a salt (compound of silver and organic acid or inorganic acid) constituting ions or silver oxides. Examples thereof include silver nitrate, silver bromide, silver oxide, silver acetate, silver carbonate, silver citrate, silver lactate, silver phosphate, silver oxalate, silver thiosulfate, silver protein, or hydrates thereof.

[0110] Examples of the buffer include Tris, sodium phosphate, potassium phosphate, histidine, boric acid buffer, and citric acid.

[0111] As a boric acid buffer, there is no particular limitation as long as it is a pharmaceutically acceptable buffer, for example, any one or two of boric acid and borates can be cited. As boric acid, there is no particular limitation as long as it is a pharmaceutically acceptable boric acid, for example, orthoboric acid, metaboric acid, tetraboric acid, etc. can be cited. As a borate, there is no particular limitation as long as it is a pharmaceutically acceptable borate, and metal salts such as sodium salts, potassium salts, calcium salts, magnesium salts, and aluminum salts can be cited; organic amine salts such as triethylamine, triethanolamine, morpholine, piperazine, and pyrrolidine can be cited. Boric acid or its salt can be used alone or in combination of two or more. In addition, as a suitable embodiment of the boric acid buffer, a combination of boric acid and borax can be cited.

[0112] When using boric acid and borax in combination, the ratio of boric acid to borax is not particularly limited. For example, the ratio of borax to 100 parts by mass of boric acid is 10 to 300 parts by mass, preferably 10 to 250 parts by mass, more preferably 30 to 100 parts by mass, and particularly preferably 40 to 60 parts by mass.

[0113] Examples of the preservative include benzalkonium chloride, methylparaben, propylparaben, chlorobutanol, sorbic acid, and alkylpolyaminoethylglycine.

[0114] Examples of the stabilizer include edetate sodium hydrate and polyvinyl pyrrolidone (povidone).

[0115] Examples of the isotonic agent include conventional sodium chloride, glycerin, glucose, mannitol, sorbitol, and sucrose, among which sucrose is preferred.

[0116] Examples of the antioxidant include ascorbic acid, ascorbic acid salts such as sodium ascorbate, tocopherol, sodium sulfite, potassium sulfite, magnesium sulfite, calcium sulfite, sodium hydrogen sulfite, potassium hydrogen sulfite, magnesium hydrogen sulfite, calcium hydrogen sulfite, sodium pyrosulfite, potassium pyrosulfite, calcium pyrosulfite, sodium thiosulfate, and sulfites such as sodium hydrogen sulfite.

[0117] As a viscosity modifier, a cellulose derivative, polyethylene glycol (Macrogol), etc. may be contained.

[0118] The pharmaceutical composition of this embodiment is not particularly limited as long as it is within the range of drugs allowed as artificial tears, and may contain aminoethanesulfonic acid, sodium chondroitin sulfate, potassium L-aspartate, magnesium L-aspartate, potassium L-aspartate / magnesium (equal mixture), sodium bicarbonate, sodium carbonate, potassium chloride, glucose, and propylene glycol.

[0119] The pharmaceutical composition of this embodiment may be a pharmaceutical preparation, and the preparation may be in the form of a solid, semisolid, or liquid. In the case of a solid, the form may be powder, granules, pills, pellets, tablets, capsules, etc. Among them, a freeze-dried powder is preferred as a solid. In the case of a semisolid, the form of a gel or the like is exemplified. In the case of a liquid, the form may be a suspension in which a powder is diluted or suspended in a buffer solution such as water or phosphate buffered saline (PBS).

[0120] For example, the usual pH range for preparing injectable drugs is about 2 to about 12. The pH of the pharmaceutical composition of some embodiments of the present invention may be in a range close to physiological pH. For example, in a specific embodiment, the pharmaceutical composition described in this specification is prepared to have a pH value in the range of pH 4 to 10 and 5 to 9.

[0121] <Method for producing pharmaceutical composition> The pharmaceutical composition of this embodiment can be produced by appropriately mixing the above-mentioned hyaluronic acid derivative and the above-mentioned active ingredient, and is preferably produced by the method described below.

[0122] That is, the method for producing a pharmaceutical composition according to the present embodiment is a method for producing a pharmaceutical composition containing a hyaluronic acid derivative and an active ingredient, and includes the following steps. A preparation step of dispersing the active ingredient in water for injection or water containing a surfactant or a polymer to prepare a dispersion containing the active ingredient; and A mixing step of selecting an arbitrary mixing ratio of the dispersion and the aqueous phase containing the hyaluronic acid derivative and mixing them.

[0123] All along, according to the ratio of the oil phase dissolving active ingredient in the organic solvent and the aqueous phase containing hyaluronic acid derivative, the oil phase can hinder the interaction between the steryl of hyaluronic acid derivative, and particles disintegrate or partially condense, etc., and are difficult to maintain the particle diameter of hyaluronic acid derivative. Therefore, there is the situation of the complex of the hyaluronic acid derivative and active ingredient of the smaller size hyaluronic acid derivative or the hyaluronic acid derivative comprising partly condensed particles after the particle disintegration in the pharmaceutical composition. In the manufacture method of the pharmaceutical composition of the present embodiment, owing to not using the oil phase, the preparation maintaining the association of hyaluronic acid derivative can be prepared. Due to the stable gelation, precipitation when being administered to the organism, it is excellent as the in situ gelation sustained-release matrix.

[0124] Next, each step of the method for producing the pharmaceutical composition according to the present embodiment will be described in detail below.

[0125] [Active ingredient microparticle preparation process] The method for producing the active ingredient can be adjusted according to the known method for purifying micron particles or nanoparticles. In this embodiment, micron or nano pulverization can be performed according to the known pulverization technology. In the present embodiment, one or more known wet crushing techniques, supercritical or compressed fluid technology, high temperature or high pressure homogenization, emulsification technology, evaporative precipitation, anti-solvent precipitation, microprecipitation, cryogenic technology, complexation technology, ultrasonic treatment technology or solid dispersion technology can be used to carry out micron or nanometer crushing. In order to separate the nanoparticles generated by aqueous or solvent dispersion technology, spray drying and freeze drying, filtration separation can be used after processing.

[0126] In this embodiment, for the method of obtaining micron or nanoparticles of an active ingredient, the active ingredient can be added to a dispersion medium to obtain a coarse dispersion, and then a dispersion of micron or nanoparticles of the active ingredient can be obtained by wet grinding; or the active ingredient can be dry-ground in advance and then added to the dispersion medium to obtain a dispersion of micron or nanoparticles of the active ingredient. The micron or nanoparticles of the active ingredient in the dispersion are preferably nanoparticles of the active ingredient obtained by wet pulverization.

[0127] The apparatus used for wet pulverization is not particularly limited, and examples thereof include a high-pressure homogenizer, a roll mill equipped with a plurality of rolls, a colloid mill, a pulverizer, a ball mill, and a bead mill.

[0128] The bead mill may be any of a batch type device, a circulation type device and a continuous type device, or a combination thereof. A batch type device refers to a device in which the entire amount of the treatment liquid is placed in a bead mill container together with a pulverizing medium and then pulverized. A circulation type device refers to a device in which the treatment liquid is circulated between a tank and a bead mill container for treatment. A continuous type device refers to a device in which the treatment liquid passes through a plurality of bead mill containers continuously.

[0129] The diameter of beads used in the bead mill is preferably 0.03 mm to 5 mm, more preferably 0.1 mm to 3 mm, further preferably 0.1 mm to 1 mm, particularly preferably 0.1 mm to 0.5 mm, and further preferably 0.1 mm to 0.3 mm.

[0130] When the diameter of the beads used in the bead mill is within the above range, the dispersion and the beads can be easily separated after the wet pulverization treatment, and the effective ingredient can be efficiently micronized.

[0131] Types of beads include glass beads, low-alkali glass beads, alkali-free glass beads, zirconia / silica ceramic beads, yttria-stabilized zirconia beads, silicon nitride beads, alumina beads, high-purity alumina beads, titania beads, etc. From the perspective of the results of use in pharmaceutical manufacturing, yttria-stabilized zirconia beads are preferred. In addition, the yttria-stabilized zirconia beads are sometimes simply referred to as zirconia beads.

[0132] When preparing a dispersion of active ingredient micron or nanoparticles, one or more selected from the group consisting of surfactants, polymers and sugars (including sugar alcohols) can be added to the dispersion medium, and the active ingredient can be added to the solution or suspension. Alternatively, after adding the active ingredient to the dispersion medium, one or more selected from the group consisting of surfactants, polymers and sugars (including sugar alcohols) can be added.

[0133] When the dispersion of active ingredient micron or nanoparticles contains either or both of a surfactant and a polymer, the mass ratio of the active ingredient to either or both of the surfactant and the polymer in the dispersion, calculated as active ingredient / surfactant / polymer, is preferably 1 / 1 / 1 to 1 / 0.01 / 0.01, and more preferably 1 / 0.75 / 0.75 to 1 / 0.1 / 0.1.

[0134] The effective ingredient may also be formulated as a eutectic of micro- or nano-particles by wet pulverization or the like.

[0135] In this embodiment, the active ingredient may be suspended in a suspension medium containing one or more carriers, excipients and diluents, or both.

[0136] From the viewpoint of ease of manufacture, micron particles are preferred. In addition, in vivo, when it is preferable not to be engulfed by phagocytes, micron particles are preferred. From the viewpoint of dissolution rate, since the surface area is small, the dissolution rate can be reduced. When the solubility of the compound is high and it is difficult to release it for a long time, there is a situation where the sustained release period can be extended by using micron particles.

[0137] From the viewpoint of dispersion stability and ease of administration, nanoparticles are preferred. Nanoparticles with smaller size and higher dispersibility are usually easier to administer than micron particles. From the viewpoint of dissolution rate, since the surface area is larger, the dissolution rate can be increased. In particular, for the purpose of increasing the dissolution rate of a compound that is poorly soluble in water, nanoparticles are preferred.

[0138] Preparation process of aqueous carrier The aqueous carrier of the present embodiment is characterized in that it contains a hyaluronic acid derivative. At this time, the hyaluronic acid derivative is dissolved in a pharmaceutically acceptable medium at any concentration for use. For example, the above-mentioned buffers such as water for injection and phosphate buffer are preferred. As a buffer, there is no particular restriction as long as it is a compound with a buffering capacity that adjusts the pH of the composition within the range of 4 to 10. As an example of a buffer, there can be cited: phosphates such as acetates such as sodium acetate, sodium dihydrogen phosphate, disodium hydrogen phosphate, potassium dihydrogen phosphate, dipotassium hydrogen phosphate, ε-aminocaproic acid, sodium glutamate, amino acid salts, boric acid and its salts, and mixtures thereof. Among them, water for injection, phosphate buffer, water for injection containing sucrose, and phosphate buffer containing sucrose are particularly preferred.

[0139] The aqueous carrier of the present embodiment may contain a pH adjuster. Examples of the pH adjuster include hydrochloric acid, citric acid, phosphoric acid, acetic acid, tartaric acid, sodium hydroxide, potassium hydroxide, sodium carbonate, and sodium bicarbonate.

[0140] If necessary, an acid such as an organic acid or an inorganic acid may be mixed. These pH adjusters may be used alone or in any combination of two or more.

[0141] The pH of the aqueous carrier of the present embodiment is not particularly limited as long as it is within a range acceptable as a drug, and is, for example, 4.0 to 9.0, preferably 4.0 to 8.8, and more preferably 6.5 to 8.8.

[0142] The aqueous carrier of the present embodiment may contain a chelating agent, and examples of the chelating agent include edetate disodium, edetate trisodium, edetate tetrasodium, diethylenetriaminepentaacetic acid, and mixtures thereof.

[0143] The aqueous carrier of the present embodiment may contain an isotonic agent, which may include conventional sodium chloride, glycerol, glucose, mannitol, sorbitol, and sucrose. The isotonic agent may be added after the active ingredient is suspended.

[0144] Other isotonic agents include glucose, lactose, mannitol, and calcium or magnesium compounds such as CaCl2. According to Japanese Patent Gazette No. 4758893, it is recorded that the antibacterial and antiseptic effects can be enhanced by mixing glycerol at a concentration (2 to 2.5% (v / v)) of substantially isotonic pressure. Therefore, not only the function as an isotonic agent, but also the antibacterial and antiseptic effects can be expected. The content of glycerol is, for example, 0.01 to 10% (w / v), preferably 0.05 to 5% (w / v), more preferably 0.1 to 3.0% (w / v), further preferably 0.3 to 3.0% (w / v), and particularly preferably 0.3 to 2.5% (w / v).

[0145] The aqueous carrier of the present embodiment may be mixed with a base as needed. There are no particular limitations on the base as long as it is pharmaceutically acceptable, and examples thereof include sodium hydroxide, potassium hydroxide, monoethanolamine, diethanolamine, triethanolamine, tromethamine, and meglumine.

[0146] The aqueous carrier of the present embodiment may be further mixed with one or more thickeners as necessary, and there are no particular limitations as long as the thickeners are pharmaceutically acceptable. Examples of thickeners for aqueous carriers in the present invention include: cellulose polymers (methyl cellulose, ethyl cellulose, hydroxyethyl cellulose, hydroxypropyl methyl cellulose, etc.), vinyl polymers (polyvinyl pyrrolidone, polyvinyl alcohol, etc.), sugars (hyaluronic acid, its salts and other mucopolysaccharides, gellan gum, sodium alginate, dextran, cyclodextrin and other polysaccharides), oxyalkylene polymers (polyoxyethylene polyoxypropylene block polymers), etc. The molecular weight of the thickener in the present invention can be, for example, from a number average molecular weight of 0.5×10 4 ~100×10 4 Select the range on the left or right.

[0147] The aqueous carrier of the present embodiment may further contain one or more preservatives as necessary, and there are no particular limitations as long as the preservatives are pharmaceutically acceptable. Examples of the preservative for the aqueous vehicle of the present invention include benzalkonium chloride, benzethonium chloride, chlorhexidine gluconate, parabens such as methylparaben / ethylparaben, benzyl alcohol, phenethyl alcohol, sorbic acid or its salts, thimerosal, and chlorobutanol.

[0148] Mixing process In the mixing step, the dispersion of the particles obtained in the preparation step and the aqueous phase containing the hyaluronic acid derivative are mixed in any proportion. For example, the mixing ratio of the dispersion of the particles and the aqueous phase containing the hyaluronic acid derivative is preferably 0.01:100 (v / v) to 100:100 (v / v). The greater the ratio of the aqueous phase containing the hyaluronic acid derivative, the more difficult it is for the active ingredient to produce a concentration gradient in the aqueous carrier, and the easier it is to disperse evenly. The greater the ratio of the dispersion of the particles, the more the dosage of the active ingredient can be increased.

[0149] In the mixing step, the concentration of the hyaluronic acid derivative dissolved in the aqueous phase is not particularly limited. The concentration of the hyaluronic acid derivative is preferably 0.01 parts by mass to 10 parts by mass, more preferably 0.1 parts by mass to 10 parts by mass, and further preferably 0.5 parts by mass to 5 parts by mass, relative to 100 parts by mass of the aqueous phase.

[0150] After the mixing step, the concentration of the hyaluronic acid derivative in the pharmaceutical composition of the present embodiment is not particularly limited. The concentration of the hyaluronic acid derivative is preferably 0.01 parts by mass to 10 parts by mass, more preferably 0.05 parts by mass to 7.5 parts by mass, and further preferably 0.3 parts by mass to 5 parts by mass, relative to 100 parts by mass of the pharmaceutical composition.

[0151] In the mixing process, the temperature is not particularly limited. From the viewpoint of reducing the viscosity of the aqueous carrier, it is preferably implemented at more than 50°C. From the viewpoint of the stability of the active ingredient, the temperature is preferably set to less than 50°C, more preferably more than 10°C and less than 55°C, and further preferably more than 15°C and less than 40°C. Alternatively, the optimal temperature can be appropriately set according to the structure and characteristics of the active ingredient. In addition, for example, it can also include a process of adjusting the temperature of the dispersion of the microparticles to 15-25°C, adjusting the temperature of the aqueous carrier containing the hyaluronic acid derivative to 40-50°C, and mixing two or more solutions with different temperatures.

[0152] The mixing step is not limited to the batch method, but the dispersion of the active ingredient and the aqueous carrier may be mixed by a flow method.

[0153] The material and shape of the reactor in the flow method may be selected as those applicable to the production method of the present invention without particular limitation. For example, the inner diameters of the pipes through which the dispersion and the aqueous carrier pass may be different.

[0154] The material and shape of the tube in the flow method can be selected without particular limitation as long as they are applicable to the method for producing the pharmaceutical composition of this embodiment. Examples thereof include tubes made of Teflon (registered trademark), stainless steel tubes, glass tubes, and plastic tubes.

[0155] In the mixing step, the time is not particularly limited, and can be, for example, 30 minutes to 90 hours, or 1 hour to 80 hours.

[0156] Drying process After the mixing step, a drying step may be performed to dry the solution containing the active ingredient and the hyaluronic acid derivative to obtain a dried product. The drying method may be the same as the method exemplified in the above-mentioned "method for producing hyaluronic acid derivatives". For example, freeze drying, spray drying, reduced pressure drying machine, and supercritical drying may be used.

[0157] <Administration Method> The subjects to whom the pharmaceutical composition of the present embodiment can be administered include mammals classified as humans (monkeys, marmosets, mice, rats, cows, horses, cats, dogs, pigs, sheep, goats, rabbits, etc.).

[0158] The administration route of the pharmaceutical composition of the present embodiment is not particularly limited, and a currently known administration route can be appropriately used depending on the intended use, the tissue site to be treated, and the like. For example, the administration route includes: subcutaneous administration, intramuscular administration, intravenous administration, intraarterial administration, intramedullary administration, intracerebral administration, intraarticular administration, intraperitoneal administration, intravaginal administration, intracapsular administration, intrarectal administration, intravitreal administration, periocular administration, intradermal administration, intraperitoneal administration, intranasal administration, transbronchial administration, pulmonary administration, transdermal administration, sublingual administration, oral administration, oral administration, and eye drop administration. Among them, subcutaneous administration, intramuscular administration, intravitreal administration, intramedullary administration, and intraarticular administration are preferred. In addition, for topical administration, it may be in the form of powder, cream, ointment or eye drops. Examples of injection administration include subcutaneous injection, intramuscular injection, intravenous injection, intraarterial injection, intramedullary injection, intraarticular injection, intraperitoneal injection, intravitreal injection, periocular injection, intradermal injection, and intratumor injection.

[0159] The administration site is preferably subcutaneous, intramuscular, intramedullary, intraarticular, intracerebral, intravitreal, ocular surface, nose, lung, and mouth. More preferably, it is subcutaneous, intramuscular, intramedullary, intraarticular, intravitreal, and ocular surface.

[0160] The administration method is preferably injection, eye drops or application.

[0161] When dripping the eye, by using hyaluronic acid derivatives, it is possible to reduce the initial burst release of the drug, suppress the coagulation that may occur between the drugs in the preparation storage and after administration, reduce the difference in drug efficacy, and then adhere to and be retained in the corneal epithelium under physiological conditions to release the drug for a longer time, reducing the number of eye drops. It is further believed that the drug can be promoted to be taken up by corneal epithelial cells, promote the drug to be taken up by conjunctival epithelial cells, etc. In addition, there is high moisture retention, viscosity, and biological adaptability derived from hyaluronic acid, and inflammation, pain, pressure, damage, etc. during contact preparation can be alleviated, and a preparation with high safety is provided. According to the above content, by effectively exerting the effect of the drug, a tear layer stabilization effect, a treatment effect of corneal epithelial disorder, a treatment effect of meibomian gland insufficiency, and an excellent pharmaceutical composition for suppressing pain effect can be provided.

[0162] When administered into the knee joint, the drug has good compatibility with hyaluronic acid present in the synovial fluid and can effectively suppress the initial burst release of the drug due to the precipitation of hyaluronic acid derivatives. Furthermore, by administering the drug together with high molecular weight hyaluronic acid (Artz, Suvenyl, etc.) used in existing drugs for OA treatment, the hyaluronic acid derivative precipitate containing the drug can be retained in the knee joint for a longer period of time.

[0163] In the pharmaceutical composition of this embodiment, when administered parenterally, the dosage can be appropriately selected in consideration of the type of the subject (including age, gender, etc.). For example, in humans (assuming a body weight of 60 kg), the amount of the active ingredient can be more than 0.01 μg and less than 100 mg, more than 0.1 μg and less than 50 mg, or more than 1 μg and less than 25 mg each time.

[0164] The administration frequency may be a single administration of the above-mentioned dose, or multiple administrations of the above-mentioned dose once every 1 week, 2 weeks, 3 weeks, 4 weeks, 1 month, 2 months, 3 months or half a year, 2 or more times, or administration to 2 or more sites in 1 administration.

[0165] <Other Implementation Methods> In one embodiment, the present invention provides a method for preventing or treating 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, bone and joint diseases, and pain diseases, comprising administering an effective amount of the above-mentioned pharmaceutical composition to a patient or animal. In addition, the "effective amount" referred to herein includes an effective amount for prevention or treatment, that is, an amount suitable for preventing or treating the onset of the above-mentioned diseases.

[0166] In one embodiment, the present invention provides a composition containing the above-mentioned active ingredient-hyaluronic acid derivative complex, which is used to prevent or treat one or more diseases selected from the group consisting of cancer, infectious diseases, immune diseases, inflammatory relaxation, allergic diseases, skin diseases, hypertension, diabetes, neurological diseases, genetic diseases, cardiovascular diseases, cerebrovascular diseases, respiratory diseases, eye diseases, ear diseases, bone and joint diseases, and pain diseases.

[0167] In one embodiment, the present invention provides a use of the above-mentioned mixture of an active ingredient and a hyaluronic acid derivative for preparing a pharmaceutical composition.

[0168] In one embodiment, the present invention provides a method for long-term sustained release of an active ingredient in vivo or in vitro, comprising administering a composition containing the active ingredient and a hyaluronic acid derivative.

[0169] One embodiment of the present invention is a pharmaceutical composition comprising an active ingredient suspended in an aqueous carrier containing a substrate that precipitates or salts out in vivo or under physiological conditions, wherein the active ingredient is a particle having a molecular weight of more than 200 g / mol and less than 5000 g / mol.

[0170] The “substrate that precipitates or salts out in vivo or under physiological conditions” is, for example, a hyaluronic acid derivative, poloxamer, unmodified chitosan or a chitosan derivative, or a peptide analog. In one embodiment of the present invention, the substrate that is precipitated or salted out in vivo or under physiological conditions is preferably a hyaluronic acid derivative, and preferably a hyaluronic acid derivative pharmaceutical composition containing an active ingredient suspended in an aqueous carrier. When hyaluronic acid derivatives are used as substrates, inflammation is not likely to occur at the administration site, and the biocompatibility is excellent. Furthermore, the ionic crosslinking produced by the carboxylic acid of the hyaluronic acid derivatives and the divalent salt in the body, and the high-density crosslinking produced between the modified steryl groups, can more stably retain the drug than other substrates, and can bring about the effects of suppressing the initial burst release and long-term sustained release.

[0171] In one embodiment of the present invention, whether or not a substrate is precipitated or salted out in vivo or under physiological conditions can be confirmed by the following method. First, the substrate dissolved to a desired concentration is added to a phosphate buffer or a phosphate buffer containing 0.8% BSA. Then, it is incubated at 37°C and centrifuged. If solid-liquid separation is possible, it can be determined that the substrate is precipitated or salted out in the physiological saline. The incubation time can be arbitrarily selected.

[0172] The concentration of the phosphate buffer is, for example, 1×PBS (about 10 mM PB, 150 mM NaCl) at the final concentration. The pH is near neutral, usually in the range of 7.1 to 7.7. The centrifugal conditions during centrifugal separation can be changed appropriately, and as an example, 9800 g is used.

[0173] In one embodiment of the present invention, when 150 μL of 4×PBS (final concentration 1×PBS) is added to 450 μL of chitosan or peptide analogs dissolved to the desired concentration, incubate under the above conditions, and further quantify the supernatant of the hyaluronic acid derivative precipitated by centrifugation by using size exclusion chromatography (SEC) or reversed phase chromatography (HPLC), the ratio of chitosan or peptide analogs precipitated or salted out under physiological conditions can be quantified. Any one of SEC or HPLC can be used for quantification, and can be appropriately selected.

[0174] In one embodiment of the present invention, when 150 μL of 4×PBS containing 3.2% BSA (1×PBS containing 0.8% BSA at a final concentration) is added to 450 μL of chitosan or peptide analogs dissolved to a desired concentration, incubation is carried out under the above conditions, and the supernatant of the hyaluronic acid derivative precipitated by centrifugation is further quantified by using size exclusion chromatography (SEC) or reverse phase chromatography (HPLC), thereby the ratio of chitosan or peptide analogs precipitated or salted out under physiological conditions can be quantified. Any one of SEC or HPLC can be used for quantification, and can be appropriately selected.

[0175] In one embodiment of the present invention, the hyaluronic acid derivative can be confirmed to precipitate or salt out under physiological saline conditions by actually administering the compound to a living body or administering the compound to a collected tissue to confirm precipitation.

[0176] The proportion of the precipitate is preferably 10 to 100% of the total amount of the dissolved hyaluronic acid derivative, preferably 40% or more, 50% or more, 60% or more, more preferably 70% or more, further preferably 80% or more, and most preferably 90% or more.

[0177] The base material precipitated or salted out in vivo or under physiological conditions gels after in vivo administration. Through this action, the active ingredient particles contained in the pharmaceutical composition are difficult to condense or diffuse. As a result, it is easy to provide a pharmaceutical composition that can stably maintain the active ingredient concentration in vivo for a long time and reduce the abnormal initial release amount (initial burst release). Example

[0178] Hereinafter, the present invention will be described in detail by way of Examples, but these are not intended to limit the scope of the present invention to the Examples.

[0179] <Example 1> The hyaluronic acid derivative pharmaceutical composition was prepared according to the following method.

[0180] [Preparation of cyclosporine A suspension] 720 mg of cyclosporin A ((manufactured by 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. Further, 10 g of 0.2 mm ZrO2 beads (manufactured by Nikkato Co., Ltd.) were added, and 14.4 mL of 1% polysorbate 80 was added. A stirring bar was added, and a magnetic stirrer was used to stir for 11 days for fine grinding. The suspension of cyclosporin A was taken out using a syringe, placed in a 15 mL conical tube, centrifuged at 10,000 g for 60 minutes, and the supernatant was discarded. Furthermore, 10 times the amount of water for injection was added, and the mixture was centrifuged at 10,000 g for 60 minutes, and the supernatant was discarded to obtain a suspension of cyclosporin. The cyclosporin suspension was diluted 200 times with a 0.05% polysorbate 80 aqueous solution, and the particle size of the nanosized cyclosporin was measured using a DLS device (ELSZ2000 manufactured by Otsuka Electronics Co., Ltd.), and the average particle size was 171 nm, and the polydispersity index was 0.12. The cyclosporin concentration was 117.7 mg / mL under the following conditions.

[0181] (Reversed Phase Chromatography Analysis Conditions) HPLC device: Japan Spectroscopic HPLC-EXTREMA HPLC quantitative conditions: reverse phase chromatography analysis conditions Chromatographic column: InertSustain C18 particle size 5μm × inner diameter 4.6mm × length 150mm Column temperature: 40°C Mobile phase: Eluent A 0.1% TFA / acetonitrile Eluent B 0.1% TFA / water Mobile phase ratio: eluent A / eluent B = 9 / 1 Flow rate: 1mL / min Injection volume: 30μL Detector: UV (210nm)

[0182] The cyclosporin suspension obtained above was vacuum dried, and the obtained powder was placed on an anti-reflection sample stand and measured under the following conditions. The obtained X-ray powder diffraction peaks are as follows: Figure 1 As shown, it is crystalline.

[0183] (XRD measurement conditions) Measuring device: Rigaku Ultima-IV X-ray source: Cu-Kα Excitation voltage: voltage 40kV, current 40mA Optical system: centralized optical system Cu-K β-ray filter: Ni foil Absorber: No Detector: Dtex (high sensitivity detector) Measurement method: θ / 2θ method Slit: DS = 1°, SS = release, RS = release, vertical slit = 10 mm 2θ / θ scanning: 2θ=2~65°(0.02° / step, 10° / min)

[0184] The cyclosporin prepared above is a nanoparticle with a molecular weight of 1202.6 g / mol and a particle size of 171 nm.

[0185] [Preparation of hyaluronic acid nanogel] As described in Examples 1 and 2 of International Publication No. 2010 / 053140, a hyaluronic acid nanogel into which a cholesteryl group is introduced (weight average molecular weight 35 kDa, cholesteryl group introduction rate 19%) is synthesized. The synthesized hyaluronic acid nanogel may be described as "HA derivative".

[0186] Phosphate buffer was added to the HA derivative dissolved to a concentration of 6 mg / mL. After that, the mixture was incubated at 37°C and centrifuged to separate the solid and liquid. This was used to determine whether the HA derivative was precipitated or salted out in the physiological saline.

[0187] The cyclosporin A suspension prepared in the above [Preparation of cyclosporin A suspension] was mixed with a 12 mg / mL aqueous solution of the HA derivative (35k19) synthesized in the above [Preparation of hyaluronic acid nanogel], and a 10% sucrose solution was further added. The hyaluronic acid derivative pharmaceutical compositions of Examples 1-1 and 1-2 were prepared in this way. The prepared samples are shown in Table 1. In the following table, "CyA concentration" represents the concentration of cyclosporin A. The final sucrose concentration was 8.7%.

[0188] [Table 1]

[0189] <Comparative Example 1-1> As Comparative Example 1-1, cyclosporin A powder was dissolved in a 10% α-cyclodextrin / 10% sucrose solution at a ratio of 0.5 mg / mL.

[0190] <Comparative Example 1-2> A 200 mg / mL cyclosporin A ethanol solution was mixed with a 12 mg / mL aqueous solution of the HA derivative (35k19) synthesized in the above [Preparation of hyaluronic acid nanogel] and stirred overnight. Sucrose was added in the form of powder as a 10% sucrose solution. The final cyclosporin concentration was 1.0 mg / mL and the 35k19 concentration was 9 mg / mL.

[0191] <Comparative Examples 1-3> The cyclosporin A suspension prepared in the above [Preparation of cyclosporin A suspension] was mixed with a 1% polysorbate 80 aqueous solution and a 10% sucrose solution. The final cyclosporin concentration was 10 mg / mL, the polysorbate 80 concentration was 0.05%, and the sucrose concentration was 8.7%.

[0192] <Pharmacokinetic Test> Each sample of Example 1-1, Example 1-2, and Comparative Examples 1-1 to 1-3 prepared in Example 1 was subcutaneously administered to normal rats (SD, 6 weeks old, male) in the dosage shown in Table 2 using a syringe with a 25G needle.

[0193] [Table 2]

[0194] After administration, blood was collected from the jugular vein using a syringe treated with heparin for a certain period of time, and EDTA-K2 was added as a protease inhibitor.

[0195] The plasma of the obtained blood was separated and measured by LC-MS / MS. The changes in the concentration of cyclosporin in the plasma when various cyclosporin suspension preparations were administered were as follows: Figure 2 and Figure 3 shown.

[0196] Depend on Figure 2 It is found that the abnormality in the initial release amount was observed in Comparative Examples 1-1 and 1-3, whereas the abnormality in the initial release amount was not observed in Examples 1-1 and 1-2. Further, by Figure 2 It was found that Example 1-1 and Example 1-2 maintained a blood concentration of 10 ng / mL or more even after 28 days from administration, and exhibited long-term sustained release.

[0197] On the other hand, in Comparative Example 1-3, the blood concentration was less than 7 ng / mL after 28 days after administration, and the long-term sustained release effect could not be confirmed. According to this result, even if the active ingredient is suspended, but not suspended in an aqueous carrier containing a hyaluronic acid derivative, the active ingredient particles diffuse in the body, so it is easy to produce abnormal initial release and cannot exert long-term sustained release.

[0198] In Comparative Example 1-1, in which the active ingredient was not suspended and the aqueous carrier did not contain a hyaluronic acid derivative, cyclosporin was not detected in the blood after 7 days. That is, almost no sustained-release effect was confirmed. According to this result, since α-cyclodextrin did not gel or precipitate at a physiological saline concentration after subcutaneous administration, it can be seen that in situ gelation and precipitation like hyaluronic acid derivatives are necessary for long-term sustained release.

[0199] In addition, Figure 3 It can be seen that in Comparative Example 1-2, in which the active ingredient and the hyaluronic acid derivative formed a complex and the active ingredient was not suspended, the initial release amount was abnormal, and the blood concentration was less than 7 ng / mL 10 days after administration, and no long-term sustained release effect was confirmed. According to the present results, when the active ingredient is not suspended in an aqueous carrier containing a hyaluronic acid derivative, the hyaluronic acid derivative undergoes in situ gelation at a physiological saline concentration, but a long-term sustained release effect is not obtained.

[0200] In the present specification, sustained release is evaluated by mean residence time (MRT (Mean Residual Time)), and more specifically, when the following formula (A) is satisfied, it is evaluated as "capable of maintaining the concentration of the active ingredient in the body for a long period of time". (MRT (time) of the hyaluronic acid derivative pharmaceutical composition) ÷ (MRT (time) of the active ingredient alone) ≥ 1.2 ···Formula (A) When the sustained release property was evaluated according to formula (A), Example 1-1 was 3.8 relative to Comparative Example 1-3; and Example 1-2 was 2.8 relative to Comparative Example 1-3, which showed that the sustained release property was obvious.

[0201] In the present specification, the abnormal initial release amount (initial burst release) is evaluated based on the maximum blood concentration Cmax, and more specifically, when the following formula (B) is satisfied, it is evaluated as "the abnormal initial release amount (initial burst release) can be reduced". {(maximum blood concentration Cmax [ng / mL] of the hyaluronic acid derivative pharmaceutical composition) ÷ (dosage of the active ingredient [mg / kg])} ÷ {(Cmax [ng / mL] of the active ingredient alone) ÷ (dosage of the active ingredient [mg / kg])} ≤ 0.8 ···Formula (B)

[0202] When the initial burst release was evaluated according to formula (B), Example 1-1 was 0.21 and Example 1-2 was 0.30 relative to Comparative Example 1-3, which shows that the abnormal initial release amount can be significantly reduced.

[0203] In the above, the following pharmacokinetic parameters were subjected to non-compartmental analysis using WinNonlin Ver.8.3 (manufactured by Pharsight). The values ​​are shown in Table 3. Mean residence time (MRT), unit: time Half-life (T1 / 2), unit: time Maximum blood concentration (Cmax), unit: ng / mL

[0204] [Table 3]

[0205] <Experimental Example 1: Sedimentation Evaluation> Using a syringe with a 30G needle, Example 1-1 and Example 1-2 prepared in Example 1 were added dropwise to a 0.8% BSA / PBS solution to form a precipitate mixed with solid cyclosporin. On the other hand, when Comparative Sample 1-3 prepared in Comparative Example 1-3 was added dropwise to the above solution, no precipitate was formed.

[0206] <Experimental Example 2: Sedimentation Evaluation> The influence of the type of hyaluronic acid derivative was verified.

[0207] [Preparation of hyaluronic acid nanogel] As described in Examples 1 and 2 of International Publication No. 2010 / 053140, a hyaluronic acid nanogel into which a cholesteryl group was introduced (weight average molecular weight: 10 kDa, introduction rate of cholesteryl group: 30%) was synthesized.

[0208] The cyclosporin A suspension prepared in Example 1 [Preparation of cyclosporin A suspension] was mixed with a 30 mg / mL aqueous solution of the HA derivative (10k30) synthesized in the above [Preparation of hyaluronic acid nanogel], and then adjusted to the concentration shown in Table 4 below using water for injection. The hyaluronic acid derivative pharmaceutical composition of Example 2-1 was prepared in this way. The prepared samples are shown in Table 4. In the following table, "CyA concentration" refers to the concentration of cyclosporin A.

[0209] [Table 4]

[0210] When Example 2-1 was added dropwise to a 0.8% BSA / PBS solution using a syringe with a 30G needle, a precipitate mixed with solid cyclosporin was formed.

[0211] <Experimental Example 3: Evaluation of precipitation in vitreous body> The vitreous humor was extracted from a pig eyeball (Cosmo Bio). Example 1-1 prepared in Example 1 was administered to the vitreous humor using a syringe. The results are shown in FIG. Figure 4 shown. like Figure 4 As shown, it was confirmed that a precipitate was formed in which cyclosporin in a solid state was mixed.

[0212] <Example 4: Intravitreal Administration> The hyaluronic acid derivative pharmaceutical composition was prepared according to the following method.

[0213] [Preparation of cyclosporine A suspension] A suspension of cyclosporin was prepared in the same manner as in Example 1. When measured using a DLS device (ELSZ2000 manufactured by Otsuka Electronics), the average particle size was 171 nm and the polydispersity index was 0.12. When the cyclosporin concentration was quantified under the following conditions, the concentration was 117.7 mg / mL.

[0214] (Reversed Phase Chromatography Analysis Conditions) HPLC device: Japan Spectroscopic HPLC-EXTREMA HPLC quantitative conditions: reverse phase chromatography analysis conditions Chromatographic column: InertSustain C18 particle size 5μm × inner diameter 4.6mm × length 150mm Column temperature: 40°C Mobile phase: Eluent A 0.1% TFA / acetonitrile Eluent B 0.1% TFA / water Mobile phase ratio: eluent A / eluent B = 9 / 1 Flow rate: 1mL / min Injection volume: 30μL Detector: UV (210nm)

[0215] [Preparation of hyaluronic acid nanogel] As described in Examples 1 and 2 of International Publication No. 2010 / 053140, a hyaluronic acid nanogel into which a cholesteryl group was introduced (weight average molecular weight: 35 kDa, introduction rate of cholesteryl group: 19%) was synthesized.

[0216] The cyclosporin A suspension prepared in the above [Preparation of cyclosporin A suspension] was mixed with a 12 mg / mL aqueous solution of the HA derivative (35k19) synthesized in the above [Preparation of hyaluronic acid nanogel] containing 10% sucrose, and a 10% sucrose solution was further added. In this way, the hyaluronic acid derivative pharmaceutical composition of Example 4-1 containing 40 mg / mL CyA was prepared. The final sucrose concentration was 6.6%.

[0217] <Comparative Example 4-1> The cyclosporin A suspension prepared in the above [Preparation of cyclosporin A suspension] was used and mixed with a 0.05% polysorbate aqueous solution at a CyA concentration of 40 mg / mL, and a 10% sucrose solution was further added to prepare Comparative Example 4-1. The final sucrose concentration was 6.1%.

[0218] <Comparative Example 4-2> A 1000 kDa hyaluronic acid (FCH-120 manufactured by KIKKOMAN) powder was weighed in a sterile medicine bottle and dissolved in a 10% sucrose aqueous solution at a concentration of 10 mg / mL. The prepared 1000 kDa hyaluronic acid (FCH-120 manufactured by KIKKOMAN) aqueous solution, the cyclosporin A suspension prepared in the above [Preparation of cyclosporin A suspension] and the 10% sucrose aqueous solution were mixed at a ratio of 40 mg / mL CyA concentration and 6.6 mg / mL hyaluronic acid concentration to prepare Comparative Example 4-2. The final sucrose concentration was 6.6%.

[0219] <Pharmacokinetic Test> Each sample of Example 4-1 and Comparative Examples 4-1 to 4-2 prepared in Example 4 was administered into the vitreous of normal rats (SD, 6 weeks old, male) at a dose of 0.8 mg / head using a syringe with a 28G needle.

[0220] [Table 5]

[0221] After administration, blood was collected from the jugular vein using a syringe treated with heparin for a certain period of time, and EDTA-K2 was added as a protease inhibitor.

[0222] The plasma of the obtained blood was separated and measured by LC-MS / MS. The concentration of cyclosporin in plasma was changed when various cyclosporin suspension preparations were administered. Figure 5 In addition, when the preparation of Example 4-1 is administered, the concentration of cyclosporine in plasma changes as shown in Figure 6 As shown, the concentration of cyclosporin in plasma changes when the preparation of Comparative Example 4-1 is administered as shown in Figure 7 As shown, the concentration of cyclosporin in plasma changes when the preparation of Comparative Example 4-2 is administered as shown in Figure 8 shown.

[0223] Depend on Figure 5 It is found that the abnormality in the initial release amount was observed in Comparative Examples 4-1 and 4-2, whereas the abnormality in the initial release amount was not observed in Example 4-1. Further, by Figures 6 to 8 The results show that in Comparative Examples 4-1 and 4-2, when n=3, CyA could not be sustained for a long time (28 days) in all groups. On the other hand, in Example 4-1, for all administration groups with n=3, there was less deviation and long-term sustained release was confirmed.

[0224] From this result, it can be seen that even if the active ingredient is suspended after administration, the initial release amount cannot be suppressed by hyaluronic acid. Even in a high-viscosity preparation such as hyaluronic acid, hyaluronic acid does not gel or precipitate at a physiological saline concentration, which shows that in situ gelation and precipitation like hyaluronic acid derivatives are necessary for long-term sustained release.

[0225] Next, the following pharmacokinetic parameters were subjected to non-compartmental analysis using WinNonlin Ver.8.3 (manufactured by Pharsight) in the same manner as in Example 1. The values ​​are shown in Table 6. Mean residence time (MRT), unit: time Half-life (T1 / 2), unit: time Maximum blood concentration (Cmax), unit: ng / mL

[0226] [Table 6]

[0227] When the sustained release was evaluated according to formula (A), Example 4-1 was 1.7 relative to Comparative Example 4-1; Example 4-1 was 2.2 relative to Comparative Example 4-2, which showed that the sustained release was obvious.

[0228] When the initial burst release was evaluated according to formula (B), Example 4-1 was 0.37 relative to Comparative Example 4-1; Example 4-1 was 0.29 relative to Comparative Example 4-2, which shows that the initial burst release can be significantly reduced.

[0229] <Example 5: Intra-articular Administration of Drugs> The same preparation as in Example 4 was used, and the hyaluronic acid derivative pharmaceutical composition of Example 5-1, the polysorbate-containing pharmaceutical composition of Comparative Example 5-1, and the hyaluronic acid-containing pharmaceutical composition of Comparative Example 5-2 were used.

[0230] <Pharmacokinetic Test> Each sample of Example 5-1 and Comparative Examples 5-1 to 5-2 prepared in Example 4 was administered into the knee joints of normal rats (SD, 6 weeks old, male) at a dose of 0.8 mg / head using a syringe with a 28G needle.

[0231] [Table 7]

[0232] After administration, blood was collected from the jugular vein using a syringe treated with heparin for a certain period of time, and EDTA-K2 was added as a protease inhibitor.

[0233] The plasma of the obtained blood was separated and measured by LC-MS / MS. The concentration of cyclosporin in plasma was changed when various cyclosporin suspension preparations were administered. Fig. 9 shown.

[0234] Depend on Figure 5 It is found that the abnormality in the initial release amount was observed in Comparative Examples 5-1 and 5-2, whereas the abnormality in the initial release amount was not observed in Example 5-1. Further, by Figures 6 to 8 The results show that in Comparative Examples 4-1 and 4-2, when n=3, CyA could not be sustained for a long time (28 days) in all groups. On the other hand, in Example 4-1, for all administration groups with n=3, there was less deviation and long-term sustained release was confirmed.

[0235] From this result, it can be seen that even if the active ingredient is suspended after administration, the initial release amount cannot be suppressed by hyaluronic acid. Even in a high-viscosity preparation such as hyaluronic acid, hyaluronic acid does not gel or precipitate at a physiological saline concentration, which shows that in situ gelation and precipitation like hyaluronic acid derivatives are necessary for long-term sustained release.

[0236] Next, the following pharmacokinetic parameters were subjected to non-compartmental analysis using WinNonlin Ver.8.3 (manufactured by Pharsight) in the same manner as in Example 1. The values ​​are shown in Table 6. Mean residence time (MRT), unit: time Half-life (T1 / 2), unit: time Maximum blood concentration (Cmax), unit: ng / mL

[0237] [Table 8]

[0238] When the sustained release was evaluated according to formula (A), Example 5-1 was 1.2 relative to Comparative Example 5-1; Example 5-1 was 1.0 relative to Comparative Example 5-2. Since the sustained release was high in both cases, almost no difference was observed.

[0239] When the initial burst release was evaluated according to formula (B), Example 5-1 was 0.61 relative to Comparative Example 5-1; Example 5-1 was 0.72 relative to Comparative Example 5-2, which shows that the initial burst release can be significantly reduced.

[0240] The present results show that only the pharmaceutical composition using a hyaluronic acid derivative achieves both long-term sustained release and suppressed release in knee joint administration.

[0241] <Example 6: Subcutaneous administration> The hyaluronic acid derivative pharmaceutical composition was prepared according to the following method.

[0242] [Preparation of cyclosporine A suspension] A suspension of cyclosporin was prepared in the same manner as in Example 1. When measured using a DLS device (ELSZ2000 manufactured by Otsuka Electronics), the average particle size was 157 nm and the polydispersity index was 0.225. When the cyclosporin concentration was quantified under the following conditions, the concentration was 64.7 mg / mL.

[0243] (Reversed Phase Chromatography Analysis Conditions) HPLC device: Japan Spectroscopic HPLC-EXTREMA HPLC quantitative conditions: reverse phase chromatography analysis conditions Chromatographic column: InertSustain C18 particle size 5μm × inner diameter 4.6mm × length 150mm Column temperature: 40°C Mobile phase: Eluent A 0.1% TFA / acetonitrile Eluent B 0.1% TFA / water Mobile phase ratio: eluent A / eluent B = 9 / 1 Flow rate: 1mL / min Injection volume: 30μL Detector: UV (210nm)

[0244] [Preparation of hyaluronic acid nanogel] As described in Examples 1 and 2 of International Publication No. 2010 / 053140, a hyaluronic acid nanogel into which a cholesteryl group was introduced (weight average molecular weight: 35 kDa, introduction rate of cholesteryl group: 19%) was synthesized.

[0245] The cyclosporin A suspension prepared in the above [Preparation of cyclosporin A suspension] was mixed with a 12 mg / mL aqueous solution containing 10% sucrose of the HA derivative (35k19) synthesized in the above [Preparation of hyaluronic acid nanogel], and a 10% sucrose solution was further added. In this way, the hyaluronic acid derivative pharmaceutical composition of Example 6-1 containing a CyA concentration of 10 mg / mL was prepared. The prepared samples are shown in Table 9. The final sucrose concentration was 8.5%.

[0246] <Comparative Example 6-1> Comparative Example 6-1 was prepared by dissolving the cyclosporin A suspension prepared in [Preparation of cyclosporin A suspension] in a 10% sucrose aqueous solution containing 0.05% polysorbate at a CyA concentration of 10 mg / mL, and further adding 10% sucrose solution. The final sucrose concentration was 8.5%.

[0247] <Comparative Example 6-2> A powder of 35 kDa hyaluronic acid (manufactured by Bloomage Biotech) was weighed in a sterile medicine bottle and dissolved in a 10% sucrose aqueous solution at a concentration of 12 mg / mL. The 10% sucrose aqueous solution containing 35 kDa hyaluronic acid, the cyclosporin A suspension prepared in the above [Preparation of cyclosporin A suspension], and the 10% sucrose aqueous solution were mixed at a ratio of 10 mg / mL CyA concentration and 5 mg / mL hyaluronic acid concentration to prepare Comparative Example 6-2. The final sucrose concentration was 8.5%.

[0248] <Comparative Example 6-3> Poloxamer 338 (manufactured by Aldrich) powder was weighed in a sterile medicine bottle and dissolved in a 10% sucrose aqueous solution at a concentration of 150 mg / mL. The 10% sucrose aqueous solution containing Poloxamer 338, the cyclosporin A suspension prepared in the above [Preparation of cyclosporin A suspension], and the 10% sucrose aqueous solution were mixed at a ratio of 10 mg / mL for CyA and 50 mg / mL for Poloxamer 338, as Comparative Example 6-3. The final sucrose concentration was 8.5%.

[0249] <Pharmacokinetic Test> Each sample of Example 6-1 and Comparative Examples 6-1 to 6-3 prepared in Example 6 was subcutaneously administered to normal rats (SD, 6 weeks old, male) at a dose of 25 mg / kg using a syringe with a 25G needle.

[0250] [Table 9]

[0251] After administration, blood was collected from the jugular vein using a syringe treated with heparin for a certain period of time, and EDTA-K2 was added as a protease inhibitor.

[0252] The plasma of the obtained blood was separated and measured by LC-MS / MS. The concentration of cyclosporin in plasma was changed when various cyclosporin suspension preparations were administered. Fig.10 shown.

[0253] Depend on Fig.10 It is found that abnormalities in the initial release amount were observed in Comparative Examples 6-1, 6-2, and 6-3, whereas abnormalities in the initial release amount were not observed in Example 6-1.

[0254] As can be known from this result, even if the active ingredient is suspended after the administration, hyaluronic acid, temperature-responsive macromolecule Poloxamer can not suppress the initial release amount. The 50mg / mL Poloxamer338 used in embodiment 6-3 is the additive used in suspension formulation REKAMBYS (Janssen Pharmaceutical company), and concentration is also identical. These preparations are not precipitated under physiological saline concentration, and it can be known that gelation in situ, precipitation are necessary for taking into account both initial burst release and long-term sustained release as hyaluronic acid derivatives.

[0255] Next, the following pharmacokinetic parameters were subjected to non-compartmental analysis using WinNonlin Ver.8.3 (manufactured by Pharsight) in the same manner as in Example 1. The values ​​are shown in Table 10. Mean residence time (MRT), unit: time Half-life (T1 / 2), unit: time Maximum blood concentration (Cmax), unit: ng / mL

[0256] [Table 10]

[0257] When the sustained release is evaluated according to formula (A), Example 6-1 is 2.6 relative to Comparative Example 6-1; Example 6-1 is 0.8 relative to Comparative Example 6-2; Example 6-1 is 3.0 relative to Comparative Example 6-3. It can be seen that Example 6-1 has significant long-term sustained release relative to Comparative Example 6-1 and Comparative Example 6-3.

[0258] When the initial burst release was evaluated according to formula (B), Example 6-1 was 0.26 relative to Comparative Example 6-1; Example 6-1 was 0.29 relative to Comparative Example 6-2; Example 6-1 was 0.14 relative to Comparative Example 6-3. This shows that the use of hyaluronic acid derivatives can significantly reduce the abnormal initial release amount.

[0259] <Example 7> The hyaluronic acid derivative pharmaceutical composition of itraconazole was prepared according to the following method.

[0260] 2.5 mg of itraconazole (manufactured by Tokyo Chemical Industry Co., Ltd.) was added to a 96-well plate, and 0.05 mL of 0.01% polysorbate 80 was added. Furthermore, 200 mg of 0.2 mm ZrO2 beads (manufactured by Nikkato), add 0.05mL of 1% polysorbate 80. Put in a stirrer, stir for 3 days using a magnetic stirrer, and grind finely. Take a portion, dilute 200 times with 0.01% polysorbate 80, and when measured using a DLS device (manufactured by Otsuka Electronics, ELSZ2000), the average particle size is 371nm and the polydispersity index is 0.21. 0.05mL of the itraconazole suspension is mixed with 0.05mL of a 12mg / mL aqueous solution of the HA derivative (35k19) synthesized in Example 1 [Preparation of hyaluronic acid nanogel] to prepare a hyaluronic acid derivative pharmaceutical composition of itraconazole. When the pharmaceutical composition is added dropwise to a subcutaneous simulated fluid (PBS solution containing 0.8% BSA (bovine serum albumin)), it is confirmed that the hyaluronic acid derivative pharmaceutical composition is gelled. The next day, the precipitate could be visually confirmed, and the gel maintained its shape even when shaken.

[0261] <Theoretical Example 1> A pharmaceutical composition of a hyaluronic acid derivative of the macrocyclic polypeptide MK-0616 (manufactured by Merck, molecular weight 1616.0) can be prepared by the following method.

[0262] 720 mg of MK-0616 was weighed in a 30 mL glass vial, and 14.4 mL of 0.01% polysorbate 80 was added. 10 g of 0.2 mm ZrO2 beads (manufactured by Nikkato) were further added, and 14.4 mL of 1% polysorbate 80 was added. A stirring bar was placed, and the mixture was stirred for 3 days using a magnetic stirrer for fine grinding. The suspension of MK-0616 was taken out using a syringe, injected into a 15 mL conical tube, and centrifuged at 10,000 g for 60 minutes, and the supernatant was discarded.

[0263] Further, 10 times the amount of water for injection was added, centrifuged at 10,000g for 60 minutes, and the supernatant was discarded to obtain a suspension of MK-0616. 1.0 mL of the MK-0616 suspension was mixed with 1.0 mL of a 12 mg / mL aqueous solution of the HA derivative (35k19) synthesized in the above-mentioned Example 1 [Preparation of hyaluronic acid nanogel] to prepare a hyaluronic acid derivative pharmaceutical composition of MK-0616. It can be fully inferred that when the pharmaceutical composition is added dropwise to a subcutaneous simulated fluid (PBS solution containing 0.8% BSA (bovine serum albumin)), the hyaluronic acid derivative pharmaceutical composition undergoes gelation and precipitation.

[0264] <Theoretical Example 2> The hyaluronic acid derivative pharmaceutical composition of insulin crystals (molecular weight 5800) can be prepared according to the following method.

[0265] Crystalline insulin zinc aqueous suspension injection is prepared according to the Japanese Pharmacopoeia (14th revised edition). Crystalline insulin zinc aqueous suspension injection can also be prepared according to Galenics of Insulin, Jens Brange, Springer Verlag, 1987 instead of the method described in the Japanese Pharmacopoeia (14th revised edition). 1.0 mL of crystalline insulin zinc aqueous suspension injection is mixed with 1.0 mL of a 12 mg / mL aqueous solution of the HA derivative (35k19) synthesized in Example 1 [Preparation of hyaluronic acid nanogel] to prepare a hyaluronic acid derivative pharmaceutical composition of insulin crystals. It can be fully inferred that when the pharmaceutical composition is added dropwise to a subcutaneous simulated fluid (PBS solution containing 0.8% BSA (bovine serum albumin)), the hyaluronic acid derivative pharmaceutical composition undergoes gelation and precipitation.

[0266] <Theoretical Example 3> The hyaluronic acid derivative pharmaceutical composition of carbamazepine-saccharin cocrystal (carbamazepine molecular weight 236.3) can be prepared according to the following method.

[0267] Carbamazepine-saccharin cocrystals were prepared according to the method described in Japanese Patent Publication No. 2007-507554. 720 mg of carbamazepine-saccharin cocrystals were weighed in a 30 mL glass vial and 14.4 mL of 0.01% polysorbate 80 was added. Further, 10 g of 0.2 mm ZrO2 beads (manufactured by Nikkato) were added and 14.4 mL of 1% polysorbate 80 was added. A stirring bar was added and stirred for 3 days using a magnetic stirrer for fine grinding. The suspension of carbamazepine-saccharin cocrystals was taken out using a syringe, injected into a 15 mL conical tube, centrifuged at 10,000 g for 60 minutes, and the supernatant was discarded. Further, 10 times the amount of water for injection was added, centrifuged at 10,000 g for 60 minutes, and the supernatant was discarded to obtain a suspension of carbamazepine-saccharin. 1.0 mL of the carbamazepine-saccharin eutectic suspension was mixed with 1.0 mL of a 12 mg / mL aqueous solution of the HA derivative (35k19) synthesized in Example 1 [Preparation of hyaluronic acid nanogel] to prepare a carbamazepine-saccharin hyaluronic acid derivative pharmaceutical composition. It can be fully inferred that when the pharmaceutical composition is added dropwise to a subcutaneous simulated fluid (PBS solution containing 0.8% BSA (bovine serum albumin)), the hyaluronic acid derivative pharmaceutical composition undergoes gelation and precipitation.

[0268] <Theoretical Example 4> The hyaluronic acid derivative pharmaceutical composition of itraconazole (molecular weight 705.6) can be prepared according to the following method.

[0269] 720 mg of itraconazole (manufactured by Tokyo Chemical Industry Co., Ltd.) was weighed in a 30 mL glass medicine bottle, and 14.4 mL of 0.01% polysorbate 80 was added. 10 g of 0.2 mm ZrO2 beads (manufactured by Nikkato Co., Ltd.) were further added, and 14.4 mL of 1% polysorbate 80 was added. A stirring bar was placed, and a magnetic stirrer was used to stir for 3 days for fine grinding. The suspension of itraconazole was taken out using a syringe, injected into a 15 mL conical tube, centrifuged at 10,000 g for 60 minutes, and the supernatant was discarded. Further, 10 times the amount of water for injection was added, centrifuged at 10,000 g for 60 minutes, and the supernatant was discarded to obtain a suspension of itraconazole. 1.0 mL of the itraconazole suspension was mixed with 3.0 mL of a 40 mg / mL aqueous solution of the HA derivative (10k30) synthesized in Example 2 to prepare a hyaluronic acid derivative pharmaceutical composition of itraconazole. It is well inferred that when the pharmaceutical composition is dropped into a subcutaneous simulated fluid (PBS solution containing 0.8% BSA (bovine serum albumin)), the hyaluronic acid derivative pharmaceutical composition is gelled and precipitated.

[0270] <Theoretical Example 5> A pharmaceutical composition of a hyaluronic acid derivative of the macrocyclic polypeptide MK-0616 (manufactured by Merck, molecular weight 1616.0) can be prepared by the following method.

[0271] Weigh 720 mg of MK-0616 in a 30 mL glass vial and add 14.4 mL of 0.01% polysorbate 80. Further add 10 g of 0.2 mm ZrO2 beads (manufactured by Nikkato) and add 14.4 mL of 1% polysorbate 80. Add a stirrer and stir for 3 days with a magnetic stirrer for fine grinding. Use a syringe to take out the suspension of MK-0616, inject it into a 15 mL conical tube, centrifuge it at 10,000 g for 60 minutes, and discard the supernatant. Further, add 10 times the amount of water for injection, centrifuge it at 10,000 g for 60 minutes, discard the supernatant, and obtain a suspension of MK-0616. 1.0 mL of the MK-0616 suspension was mixed with 3.0 mL of a 40 mg / mL aqueous solution of the HA derivative (10k30) synthesized in Example 2 [Preparation of hyaluronic acid nanogel] to prepare a hyaluronic acid derivative pharmaceutical composition of MK-0616. It can be fully inferred that when the pharmaceutical composition is added dropwise to a subcutaneous simulated fluid (PBS solution containing 0.8% BSA (bovine serum albumin)), the hyaluronic acid derivative pharmaceutical composition undergoes gelation and precipitation.

[0272] <Theoretical Example 6> The hyaluronic acid derivative pharmaceutical composition of insulin crystals (molecular weight 5800) can be prepared according to the following method.

[0273] Crystalline insulin zinc suspension injection is prepared according to the Japanese Pharmacopoeia (14th revised edition). Alternatively, the method described in the Japanese Pharmacopoeia (14th revised edition) can be replaced by preparing a crystalline insulin zinc aqueous suspension injection according to Galenics of Insulin, Jens Brange, Springer Verlag, 1987. 1.0 mL of crystalline insulin zinc aqueous suspension injection is mixed with 3.0 mL of a 40 mg / mL aqueous solution of the HA derivative (10k30) synthesized in Example 2 [Preparation of hyaluronic acid nanogel] to prepare a hyaluronic acid derivative pharmaceutical composition of insulin crystals. It can be fully inferred that when the pharmaceutical composition is added dropwise to a subcutaneous simulated fluid (PBS solution containing 0.8% BSA (bovine serum albumin)), the hyaluronic acid derivative pharmaceutical composition undergoes gelation and precipitation.

[0274] <Theoretical Example 7> The hyaluronic acid derivative pharmaceutical composition of carbamazepine-saccharin cocrystal (carbamazepine molecular weight 236.3) was prepared according to the following method.

[0275] Carbamazepine-saccharin cocrystals were prepared according to the method described in Japanese Patent Publication No. 2007-507554. 720 mg of carbamazepine-saccharin cocrystals were weighed in a 30 mL glass vial and 14.4 mL of 0.01% polysorbate 80 was added. Further, 10 g of 0.2 mm ZrO2 beads (manufactured by Nikkato) were added and 14.4 mL of 1% polysorbate 80 was added. A stirring bar was added and stirred for 3 days using a magnetic stirrer for fine grinding. The suspension of carbamazepine-saccharin cocrystals was taken out using a syringe, injected into a 15 mL conical tube, centrifuged at 10,000 g for 60 minutes, and the supernatant was discarded. Further, 10 times the amount of water for injection was added, centrifuged at 10,000 g for 60 minutes, and the supernatant was discarded to obtain a suspension of carbamazepine-saccharin. 1.0 mL of the carbamazepine-saccharin eutectic suspension was mixed with 3.0 mL of a 40 mg / mL aqueous solution of the HA derivative (10k30) synthesized in Example 2 [Preparation of hyaluronic acid nanogel] to prepare a carbamazepine-saccharin hyaluronic acid derivative pharmaceutical composition. It can be fully inferred that when the pharmaceutical composition is added dropwise to a subcutaneous simulated fluid (PBS solution containing 0.8% BSA (bovine serum albumin)), the hyaluronic acid derivative pharmaceutical composition undergoes gelation and precipitation.

[0276] <Theoretical Example 8> A pharmaceutical composition of a hyaluronic acid derivative of acetaminophen (manufactured by Tokyo Chemical Industry Co., Ltd., molecular weight 151.2) can be prepared by the following method.

[0277] 720 mg of acetaminophen was weighed in a 30 mL glass vial, and 14.4 mL of 0.01% polysorbate 80 was added. 10 g of 0.2 mm ZrO2 beads (manufactured by Nikkato) were further added, and 14.4 mL of 1% polysorbate 80 was added. A stirring bar was placed, and a magnetic stirrer was used to stir for 3 days and finely pulverize. The suspension of acetaminophen was taken out using a syringe, injected into a 15 mL conical tube, and centrifuged at 10,000 g for 60 minutes, and the supernatant was discarded.

[0278] Further, add 10 times the amount of water for injection, centrifuge at 10,000g for 60 minutes, discard the supernatant, and prepare the suspension of paracetamol. 1.0mL of this paracetamol suspension is mixed with 1.0mL of the 12mg / mL aqueous solution of the HA derivative (35k19) synthesized in the above-mentioned embodiment 1 [preparation of hyaluronic acid nanogel] to manufacture the hyaluronic acid derivative pharmaceutical composition of paracetamol. It can be fully inferred that when the pharmaceutical composition is added dropwise to the subcutaneous simulated fluid (PBS solution containing 0.8% BSA (bovine serum albumin)), the hyaluronic acid derivative pharmaceutical composition is gelled and precipitated.

[0279] <Theoretical Example 9> A pharmaceutical composition of a hyaluronic acid derivative of acetaminophen (manufactured by Tokyo Chemical Industry Co., Ltd., molecular weight 151.2) can be prepared by the following method.

[0280] 720 mg of acetaminophen was weighed in a 30 mL glass vial, and 14.4 mL of 0.01% polysorbate 80 was added. 10 g of 0.2 mm ZrO2 beads (manufactured by Nikkato) were further added, and 14.4 mL of 1% polysorbate 80 was added. A stirring bar was placed, and a magnetic stirrer was used to stir for 3 days and finely pulverize. The suspension of acetaminophen was taken out using a syringe, injected into a 15 mL conical tube, and centrifuged at 10,000 g for 60 minutes, and the supernatant was discarded.

[0281] Further, add 10 times the amount of water for injection, centrifuge at 10,000g for 60 minutes, discard the supernatant, and make a suspension of acetaminophen. 1.0mL of this acetaminophen suspension is mixed with 3.0mL of the 40mg / mL aqueous solution of the HA derivative (10k30) synthesized in the above-mentioned embodiment 2 [preparation of hyaluronic acid nanogel] to manufacture the hyaluronic acid derivative pharmaceutical composition of acetaminophen. It can be fully inferred that when the pharmaceutical composition is added dropwise to a subcutaneous simulated fluid (PBS solution containing 0.8% BSA (bovine serum albumin)), gelation and precipitation occur in the hyaluronic acid derivative pharmaceutical composition. Industrial Applicability

[0282] According to the hyaluronic acid derivative pharmaceutical composition of this embodiment, it is possible to provide a hyaluronic acid derivative pharmaceutical composition which can maintain the concentration of the active ingredient in the body for a long period of time and reduce the abnormal initial release amount (initial burst release).

Claims

1. A hyaluronic acid derivative pharmaceutical composition, comprising an active ingredient suspended in an aqueous carrier, wherein the aqueous carrier contains a hyaluronic acid derivative, and the active ingredient is a particle with a molecular weight of more than 100 g / mol and less than 8000 g / mol. 2 . The hyaluronic acid derivative pharmaceutical composition according to claim 1 , wherein the microparticles are microparticles or nanoparticles.

3. The hyaluronic acid derivative pharmaceutical composition according to claim 1 or 2, wherein the hyaluronic acid derivative contains one or more repeating units represented by the following general formula (I): In the formula, R 1 , R 2 , R 3 and R 4 Each independently consists of hydrogen atoms, C 1-6 Alkyl, formyl and C 1-6 A group selected from the group consisting of alkylcarbonyl, Z represents a direct bond or a peptide linker consisting of 2 or more and 30 or less arbitrary amino acid residues; X 1 For-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 A group selected from the group consisting of groups represented by -SR and -SSR; R a , R b and R c Each independently consists of hydrogen atoms, C 1-20 Alkyl, amino C 2-20 Alkyl and hydroxyl C 2-20 A group selected from the group consisting of alkyl; R a , R b and R c The alkyl moiety is inserted by -O- and -NR f -Selected groups in the constituted group or not inserted; R f Composed of hydrogen atoms, C 1-12 Alkyl, amino C 2-12 Alkyl and hydroxyl C 2-12 A group selected from the group consisting of alkyl groups, R f The alkyl portion is inserted with a group selected from the group consisting of -O- and -NH- or is not inserted; R is a hydrophobic group or a hydrophilic group; Y is C 2-30 Alkylene or -(CH2CH2O) m -CH2CH2-, where The alkylene insertion of Y is -O-, -NR g - and -SS- is a group selected from the group consisting of or not inserted; R g Composed of hydrogen atoms, C 1-20 Alkyl, amino C 2-20 Alkyl and hydroxyl C 2-20 A group selected from the group consisting of alkyl groups, R g The alkyl portion is inserted with a group selected from the group consisting of -O- and -NH- or is not inserted; Y a C 1-5 Alkylene; Y b C 2-8 Alkylene or C 2-8 Alkenylene, m is an integer of 1 to 100 both inclusive. The hyaluronic acid derivative pharmaceutical composition according to claim 3 , wherein R is a hydrophobic group. The hyaluronic acid derivative pharmaceutical composition according to claim 4 , wherein the hydrophobic group is a cholesteryl group.

6. The hyaluronic acid derivative pharmaceutical composition according to claim 1 or 2, wherein the hyaluronic acid derivative is precipitated or salted out in vivo or under physiological conditions.

7. The hyaluronic acid derivative pharmaceutical composition according to claim 1 or 2, wherein the active ingredient is a poorly water-soluble drug.

8. The hyaluronic acid derivative pharmaceutical composition according to claim 1 or 2, wherein the active ingredient is a cyclic polypeptide. 9 . The hyaluronic acid derivative pharmaceutical composition according to claim 8 , wherein the cyclic polypeptide comprises a ring size of 4 to 49 amino acids. 10 . The hyaluronic acid derivative pharmaceutical composition according to claim 8 , wherein the cyclic polypeptide comprises a non-natural amino acid. 11 . The pharmaceutical composition of hyaluronic acid derivatives according to claim 8 , wherein the cyclic polypeptide is composed of 8 or more and 20 or less amino acids.

12. The hyaluronic acid derivative pharmaceutical composition according to claim 1 or 2, wherein the administration site is subcutaneous, intramuscular, intramedullary, intraarticular, intracerebral, intravitreal, ocular surface, nose, lung, or mouth.

13. The hyaluronic acid derivative pharmaceutical composition according to claim 1 or 2, wherein the administration method is injection, eye drops or coating.

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