Novel lipid accumulation inhibitor and therapeutic agent for lipid storage disease containing same

Abstract

The object of the present invention is to provide a novel pharmaceutical composition for use in the treatment or prevention of lysosomal diseases, particularly Niemann-Pick disease. According to the present invention, there is provided a pharmaceutical composition for the treatment or prevention of a lysosomal disease, comprising as an active ingredient a modified γ-cyclodextrin in which a monovalent group derived from galacturonic acid is bonded to a sugar residue of the γ-cyclodextrin via an amide bond.

Description

NOVEL LIPID ACCUMULATION INHIBITOR AND THERAPEUTIC AGENT FOR LIPID STORAGE DISEASE CONTAINING SAME

[0001] The present invention relates to a novel inhibitor of lipid accumulation, particularly cholesterol accumulation. The present invention also relates to a therapeutic agent which comprises the lipid accumulation inhibitor for a disease associated with lipid accumulation-related diseases, particularly cholesterol accumulation-related diseases.

[0002] When an enzyme associated with lysosomes, which are one of the intracellular organelles, is genetically deficient or mutated, substances that should normally be degraded or transported accumulate intracellularly as foreign bodies. Congenital metabolic disorders caused by such phenomena are known as lysosomal diseases. Lysosomal diseases have different names and symptoms depending on the enzyme that is deficient, and approximately 50 types of diseases are currently known. Examples of lysosomal diseases include cholesterol storage diseases and sphingolipidoses, which are characterized by the accumulation of lipids called sphingolipids, including Niemann-Pick disease, Gaucher disease, GM1 gangliosidosis, GM2 gangliosidosis, Fabry disease, and the like.

[0003] Niemann-Pick disease type C (NPC) is one of the congenital lysosomal diseases caused by abnormalities in NPC1, a membrane protein responsible for intracellular lipid transport, primarily cholesterol, or NPC2, a secretory protein coexisting with NPC1 in endosomes. In Niemann-Pick disease type C (NPC), deficiency of the cholesterol transport proteins NPC1 and NPC2 leads to the accumulation of free cholesterol and the like in the central nervous system and organs. Patients develop neurological disorders (ataxia, seizures, and psychomotor developmental delay) and hepatic impairment in early childhood, resulting in death by around age 20. The approved drug Miglustat has limited efficacy and is ineffective against cholesterol accumulation. No other effective therapeutic drugs are available.

[0004] Clinical trials of 2-hydroxypropyl-β-cyclodextrin (HPBCD) are underway in Europe and the United States. However, HPBCD is known to be ototoxic, which hampers its application to patients (Non-Patent Literature 1). Furthermore, in Japan, HPBCD has been associated with severe lung damage, and systemic administration has been discontinued. Therefore, there is an urgent need to develop a drug with excellent efficacy and safety.

[0005] The present inventors have previously found and reported that 2-hydroxypropyl-γ-cyclodextrin (HPGCD) has cholesterol accumulation-reducing effect equivalent to HPBCD, but has milder side effects than HPBCD (Non-Patent Literature 2, Patent Literature 1). However, HPGCD still has auditory toxicity (ototoxicity), although it is much weaker than HPBCD (Non-Patent Literature 3).

[0006] Furthermore, cyclodextrin derivatives have been reported in which the hydroxyl group at the 6-position of the sugar residues of γ-cyclodextrin is linked to the anomeric hydroxyl group of glucose or maltose via a glucosidic bond (the 6-position carbon of a sugar residue of cyclodextrin is linked to the 1-position carbon of the glucosyl or maltosyl group via -O-) (Non-Patent Literature 4). It was reported that γ-cyclodextrin derivatives linked to a glucosyl or maltosyl group exhibited significantly reduced ototoxicity when administered subcutaneously to mice compared with HPBCD or HPGCD, but retained equivalent ototoxicity when administered intracerebroventricularly (Non-Patent Literature 4).

[0007] GM1 gangliosidosis is caused by mutations in lysosomal β-galactosidase, a glycoside hydrolase. GM1 gangliosidosis is a disease in which deficiency of β-galactosidase leads to the accumulation of glycolipids such as GM1-ganglioside and asialo GM1-ganglioside, which are its substrates, in the brain, visceral organs (liver and spleen) or the like, and to the accumulation of mucopolysaccharides such as keratan sulfate in the bones. There are three types of GM1 gangliosidosis: infantile type (type 1), which develops in early infancy and is characterized by widespread central nervous system disorders including spastic paraplegia, cherry-red spots on the fundus, hepatosplenomegaly, and bone abnormalities; juvenile type (type 2), which develops in early childhood and progresses with central nervous system disorders; and adult type (type 3), which develops in school-age and presents with symptoms such as dysarthria and is characterized primarily by extrapyramidal symptoms. GM2 gangliosidosis is caused by the accumulation of a lipid called GM2 ganglioside in nerve cells due to a deficiency of hexosaminidase A, a glycoside hydrolase. Tay-Sachs disease and Sandhoff disease fall into this category.

[0008] Enzyme replacement therapy has been the main treatment for these diseases to date, but there are problems including the fact that enzyme preparations do not easily reach the central nervous system, making them ineffective in treating the nervous system including the brain, and the need for lifelong intravenous infusion of expensive enzyme preparations. Therefore, novel therapeutic drugs for these lysosomal diseases are desired.

[0009] Various cyclodextrin derivatives have been developed for the purposes of improving the solubility of cyclodextrin in water or organic solvents, making it insoluble in water, modifying polymer surfaces, adding new properties, and the like. Among these, branched cyclodextrins in which sugars are attached to cyclodextrin in a branched structure are known. Specifically, branched cyclodextrins with branched structures derived from sugars such as glucose, maltose, maltooligosaccharide, galactose, and mannose are known (Non-Patent Literatures 5 to 7, Patent Literatures 2 to 5). These branched cyclodextrins are synthesized by enzymatic reactions, and because a glycosidic bond is formed by dehydration condensation between a hydroxyl group of one of the glucose residues constituting the cyclodextrin and the hydroxyl group attached to the 1-position carbon of the above sugars, and thus none of them possesses reducing properties.

[0010] Furthermore, novel cyclodextrin derivatives having a branched structure in which an amino group of aminated α-cyclodextrin, β-cyclodextrin, or γ-cyclodextrin is bonded to the carboxyl group of uronic acid via an amide bond, thereby having a branched structure derived from uronic acid in the cyclodextrin and possessing reducing properties, and a method for producing the same, have also been reported (Patent Literature 6).

[0011] WO2015 / 083736JP2020-176063AJP61-92592AJP10-36406AJP08-107794AJP2020-37673A

[0012] Daniel S. Ory et al., Intrathecal 2-hydroxypropyl-β-cyclodextrin decreases neurological disease progression in Niemann-Pick disease type C1: a non-randomised, open-label, phase 1-2 trial, Lancet, 2017 Oct 14; 390(10104):1758-1768.Soga et al., HPGCD outperforms HPBCD as a potential treatment for Niemann-Pick disease type C during disease modeling with iPS cells, Stem Cells, 2015 Apr; 33(4):1075-88.Cristin D. Davidson et al., Efficacy and ototoxicity of different cyclodextrins in Niemann-Pick C disease, Ann Clin Transl Neurol., 2016 Apr 20; 3(5):366-80.Yamada et al., Different solubilizing ability of cyclodextrin derivatives for cholesterol in Niemann-Pick disease type C treatment, Clin Transl Med., 2023 Aug; 13(8):e1350.Application Technology of Cyclodextrins, CMC Publishing Co., Ltd., February 2008, pp. 262-268.Encyclopedia of Starch Science, Asakura Publishing Co., Ltd., March 2003, pp. 479-483.Oligosaccharides I - Starch-related Oligosaccharides, Japan Confectionery and Innovative Food Ingredients Research Center, March 2015, pp. 61-76.

[0013] An object of the present invention is to provide a novel lipid accumulation inhibitor and / or a therapeutic agent for lipid storage disease, which has an effect of reducing accumulation of lipid, particularly cholesterol, equal to or greater than that of 2-hydroxypropyl-γ-cyclodextrin (HPGCD), reported by the present inventors, and which has fewer side effects than HPGCD.

[0014] As a result of extensive research into cyclodextrin derivatives, the present inventors have discovered reduced cyclodextrins having a group derived from galacturonic acid (referred to as "RGalγ-CD" or "GalGCD") (RGal6γCD and RGal3γCD, in which groups derived from galacturonic acid are attached to the 6-position and 3-position of the sugar) and having the cholesterol accumulation-reducing effect equivalent to HPGCD but with fewer side effects, thereby completing the present invention. The present invention includes the following. [1] A pharmaceutical composition for treating or preventing a lysosomal disease, comprising a modified γ-cyclodextrin as an active ingredient, wherein in the modified γ-cyclodextrin, a monovalent group derived from galacturonic acid is bonded to a sugar residue of the γ-cyclodextrin via an amide bond. [2] The pharmaceutical composition according to [1], wherein in the modified γ-cyclodextrin, a monovalent group derived from galacturonic acid is bonded to the 6-position carbon and / or the 3-posirion carbon of a sugar residue of the γ-cyclodextrin via an amide bond. [3] The pharmaceutical composition according to [1], wherein the modified γ-cyclodextrin is a compound represented by the following formula (1):

[0015] (In the formula, m is 0 to 7, n is 0 to 8, o is 0 to 8, and m+n+o=8, and n R1and R2groups each independently represent a hydroxyl group or the following formula (a): -NH-Z (a) [wherein Z represents a monovalent group formed by removing a hydroxy group (-OH) from the carboxy group (-COOH) of galacturonic acid], and o R3, R4and R5groups each independently represent a hydroxy group or the formula (a), preferably o R3and R4groups each independently represent a hydroxyl group or the formula (a), and o R5group represents the formula (a). here, in the formula (1), when n+o is 2 or more, the arrangement of structures (A), (B), and (C) below is arbitrary, and n (B)s may be the same or different, and o (C)s may be the same or different.)

[0016] . [4] The pharmaceutical composition according to [1], wherein the modified γ-cyclodextrin is a compound represented by the following formula (2):

[0017] (In the formula, m is 0 to 7, n is 1 to 8, and m+n=8, and n R1and R2groups each independently represent a hydroxyl group the following formula (a): -NH-Z (a) [wherein Z represents a monovalent group formed by removing the hydroxy group (-OH) from the carboxy group (-COOH) of galacturonic acid.], here, in formula (2), when n is 2 or more, the arrangement of structure (A) and (B) below is arbitrary, and n Bs may be the same or different.)

[0018] or by the following formula (3):

[0019] (In the formula, m is 0 to 7, o is 1 to 8, and m+o=8, and o R3, R4and R5groups each independently represent a hydroxy group or the following formula (a): -NH-Z (a) [wherein, Z represents a monovalent group formed by removing the hydroxy group (-OH) from the carboxy group (-COOH) of galacturonic acid.], provided that not all of R3, R4, and R5are hydroxy groups, preferably o R3and R4groups each independently represent a hydroxyl group or the formula (a), and o R5group represents the formula (a), here, in formula (3), when o is 2 or more, the arrangement of structure (A) and (C) below is arbitrary, and o (C)s each may be the same or different.)

[0020] . [5] The pharmaceutical composition according to [4], wherein the modified γ-cyclodextrin is represented by the formula (2), wherein n is 1, R1is the formula (a), and R2is OH. [6] The pharmaceutical composition according to [4], wherein the modified γ-cyclodextrin is represented by the formula (3), wherein o is 1, R3and R4are OH, and R5is the formula (a). [7] The pharmaceutical composition according to any one of [1] to [6], wherein the lysosomal disease is a lipid storage disease (e.g., cholesterol storage disease, sphingolipidosis, or lysosomal acid lipase deficiency). [8] The pharmaceutical composition according to [7], wherein the lipid storage disease is selected from the group consisting of Niemann-Pick disease type C, GM1 gangliosidosis, and GM2 gangliosidosis. [9] The pharmaceutical composition according to any one of [1] to [8], wherein the pharmaceutical composition is administered parenterally.

[0010] The pharmaceutical composition according to any one of [1] to [8], wherein the pharmaceutical composition is administered as an injection.

[0021]

[0011] A method for treating or preventing a lysosomal disease, comprising administering to a subject in need thereof a therapeutically effective amount of a modified γ-cyclodextrin in which a monovalent group derived from galacturonic acid is bonded to a sugar residue of γ-cyclodextrin via an amide bond.

[0012] The method according to

[0011] , wherein the modified γ-cyclodextrin is one in which a monovalent group derived from galacturonic acid is bonded via an amide bond to the 6-position carbon and / or the 3-position carbon of a sugar residue of γ-cyclodextrin.

[0013] The method according to

[0011] , wherein the modified γ-cyclodextrin is represented by the formula (1) (the definitions in the formula are the same as above).

[0014] The method according to

[0013] , wherein the modified γ-cyclodextrin is represented by the formula (2) (the definitions in the formula are the same as above) or the formula (3) (the definitions in the formula are the same as above).

[0015] The method according to

[0013] , wherein the modified γ-cyclodextrin is represented by the formula (2), in which n is 1, R1is the formula (a), and R2is OH.

[0016] The method according to

[0013] , wherein the modified γ-cyclodextrin is represented by the formula (3), in which o is 1, R3and R4are OH, and R5is the formula (a).

[0017] The method according to any one of

[0011] to

[0016] , wherein the lysosomal disease is a lipid storage disease (e.g., cholesterol storage disease, sphingolipidosis, or lysosomal acid lipase deficiency, particularly Niemann-Pick disease, Gaucher disease, GM1 gangliosidosis, GM2 gangliosidosis, or Fabry disease).

[0018] The method according to any one of

[0011] to

[0016] , wherein the lysosomal disease is selected from the group consisting of Niemann-Pick disease type C, GM1 gangliosidosis, and GM2 gangliosidosis.Advantageous Effect of Invention

[0022] The pharmaceutical composition of the present invention, which contains as an active ingredient, γ-cyclodextrin in which a monovalent group derived from galacturonic acid is amide-bonded to a sugar residue of γ-cyclodextrin (GalGCD), is effective in treating or preventing lysosomal diseases, particularly lipid storage diseases (e.g., Niemann-Pick disease, GM1 gangliosidosis, and GM2 gangliosidosis). GalGCD contained in the pharmaceutical composition of the present invention is useful because it has lower ototoxicity than conventional cyclodextrin derivatives such as HPBCD and HPGCD.

[0023] Figure 1 shows the results of measuring the total protein mass when various concentrations of γ-cyclodextrin derivatives were added. From the left, the results are for Gal6GCD, Gal3GCD, Glc6GCD, and Glc3GCD. The same applies to Figures 2 to 4. Figure 2 is a graph showing the results of the concentration-dependent response of GalGCD addition on the effect of improving intracellular cholesterol (total cholesterol) accumulation in Npc1-deficient CHO cells. Data represent mean ± SEM (N = 3), with * indicating p < 0.05 (compared to wild-type (WT)), and # indicating p < 0.05 (compared to Npc1 null). The upper row shows the total cholesterol accumulation rate in nmol / mg protein, and the lower row shows the rate as a percentage relative to WT set at 100%. Figure 3 is a graph showing the results of the concentration-dependent response of GalGCD addition on the effect of reducing intracellular cholesterol (free cholesterol) accumulation in Npc1-deficient CHO cells. Data represent mean ± SEM (N = 3), with * indicating p < 0.05 (compared to WT) and # indicating p < 0.05 (compared to Npc1 null). The upper row shows the free cholesterol accumulation rate in nmol / mg protein, and the lower row shows the rate as a percentage relative to WT set at 100%. Figure 4 is a graph showing the results of the concentration-dependent response of GalGCD addition on the effect of improving intracellular cholesterol accumulation (ratio of total cholesterol to esterified cholesterol) in Npc1-deficient CHO cells. Data represent mean ± SEM (N = 3), with * indicating p < 0.05 (compared to WT) and # indicating p < 0.05 (compared to Npc1 null). The upper row shows the ratio of total cholesterol to esterified cholesterol, and the lower row shows the percentage relative to WT set at 100%. Figure 5 shows the results of the concentration-dependent response of GalGCD addition on the effect of improving intracellular free cholesterol accumulation in neural stem cells differentiated from NPC-derived iPS cells. Data represent mean ± SEM (N = 3), with * representing p < 0.05, ** representing p < 0.01, and *** representing p < 0.001 (compared to compound-untreated NPCs). Panel A shows the reducing effect of GalGCD on cholesterol accumulation, visualized by Filipin staining of free cholesterol. Panel B is a graph of the degree of reduction in panel A. Figure 6 shows the results of evaluating the effect of GalGCD on reducing LC3B-II protein expression (A) and insoluble p62 (B) using neural stem cells induced from the NPC-iPS cell line. A and B are the results of Western blotting, and C is a graph showing protein expression levels after normalization to tubulin expression. Normal indicates neural stem cells induced from an iPS cell line derived from a normal individual, and NPC indicates neural stem cells induced from an iPS cell line derived from an NPC patient. “-“ indicates no addition of GalGCD or HPGCD. Figure 7 shows the results of measuring auditory brainstem response (ABR) after repeated intracerebroventricular administration of GalGCD (A) and HPGCD (B) to normal adult mice. Each dot represents the result of an individual mouse.

[0024] The present invention will be described below, taking exemplary embodiments as examples, along with preferred methods and materials that can be used in carrying out the present invention, but the present invention is not limited to the embodiments described below. Unless otherwise specified, all technical and scientific terms used herein have the same meaning as commonly understood by those skilled in the art to which the present invention belongs. Furthermore, any materials and methods equivalent or similar to those described herein can also be used in carrying out the present invention. Furthermore, all publications and patents cited in this specification in connection with the present invention are incorporated herein by reference and constitute a part of this specification, for example, to describe methods, materials, and the like that can be used in the present invention.

[0025] In this specification, the expression "A to B" for a numerical range means a numerical range including the endpoints A and B. The same applies to "A through B". In addition, in this specification, the term "about" is used to mean a tolerance of ±10%.

[0026] 1. Modified γ-cyclodextrin The pharmaceutical composition of the present invention contains as an active ingredient, γ-cyclodextrin in which a monovalent group derived from galacturonic acid is amide-bonded to a sugar residue of γ-cyclodextrin (GalGCD). GalGCD used in the present invention is a compound represented by the following formula (1).

[0027]

[0028] In the formula (1), m is 0 to 7, n is 0 to 8, o is 0 to 8, and m+n+o=8. That is, at least either n or o is 1 or greater. n R1and R2groups each independently represent a hydroxyl group or the following formula (a): -NH-Z (a). In formula (a), Z represents a monovalent group formed by removing the hydroxy group (-OH) from the carboxy group (-COOH) of galacturonic acid. o R3, R4and R5groups each independently represent a hydroxyl group or the formula (a). It is preferable that o R3and R4groups each independently represent a hydroxyl group or the formula (a), and o R5group represents the formula (a). In addition, in formula (1), when n+o is 2 or more, the structures (A), (B), and (C) below may be arranged arbitrarily, and n (B)s may be the same or different, and o (C)s may be the same or different.

[0029]

[0030] In one embodiment, the GalGCD of the present invention is a compound represented by the following formula (2):

[0031] In the formula (2), m is 0 to 7, n is 1 to 8, and m+n=8. n R1and R2groups each independently represent a hydroxyl group or the following formula (a): -NH-Z (a) In formula (a), Z represents a monovalent group formed by removing the hydroxy group (-OH) from the carboxy group (-COOH) of galacturonic acid. Here, in formula (2), when n is 2 or more, the structure (A) and (B) below may be arranged arbitrarily, and the n (B)s may be the same or different.

[0032]

[0033] In one preferred embodiment, the GalGCD of the present invention is represented by the formula (2), in which n is 1, R1is the formula (a), and R2is OH.

[0034] In one embodiment, the GalGCD of the present invention is a compound represented by the following formula (3):

[0035] In the formula (3), m is 0 to 7, o is 1 to 8, and m+o=8. o R3, R4and R5groups each independently represent a hydroxyl group or a group represented by the following formula (a): -NH-Z (a) In formula (a), Z represents a monovalent group formed by removing the hydroxy group (-OH) from the carboxy group (-COOH) of galacturonic acid. It is preferable that o R3and R4groups each independently represent a hydroxyl group or the formula (a), and o R5group represents the formula (a). Here, in formula (3), when o is 2 or more, the structures (A) and (C) below may be arranged arbitrarily, and the o (C)s may be the same or different.

[0036]

[0037] In one preferred embodiment, the GalGCD of the present invention is represented by the formula (3), in which o is 1, R3and R4are OH, and R5is the formula (a).

[0038] In the GalGCD used in the present invention, the group represented by formula (a) is bonded to the 6-position and / or 3-position and / or 2 of a sugar residue of γ-cyclodextrin, and one, two, or three groups represented by formula (a) may be bonded to the same sugar residue. In the GalGCD of the present invention, the group represented by formula (a) may be present in one sugar residue of cyclodextrin, or in a plurality of sugar residues, or even in all eight sugar residues. When two or more groups represented by formula (a) are present in the GalGCD, the bonding positions of the groups represented by formula (a) to the sugar residues are selected from 6-position, 3-position and 2-position, and these positions may be the same or different. For example, when three groups represented by formula (a) are present in the GalGCD, the bonding positions thereof to the sugar residues may all be the same, two may be the same, or all may be different. Furthermore, when two or more sugar residues having a group represented by formula (a) bonded thereto are present in GalGCD, the positions of these sugar residues within the GCD having eight sugar residues are arbitrary.

[0039] The GalGCD used in the present invention has the group represented by the formula (a) preferably at 6-position or 3-position, more preferably at 6-position.

[0040] In the GalGCD used in the present invention, in formula (1) above, n+o is 1 to 8, preferably 1 to 3, more preferably 1 or 2, and even more preferably 1. For example, a GalGCD in which n is 1, o is 0, and the formula (a) is bonded to the 6-position carbon is represented by the following formula (4).

[0041]

[0042] The GalGCD, in which n is 0, o is 1, and the formula (a) is bonded to the 3-positiion carbon is represented by the following formula (5).

[0043]

[0044] The GalGCD of the present invention can be produced by condensing, in the presence of a condensing agent, the carboxyl group of galacturonic acid with the amino group of amino-modified γ-cyclodextrin (aminated GCD) obtained by substituting a hydroxyl group of a sugar residue of γ-cyclodextrin (GCD) with an amino group. That is, the GalGCD of the present invention can be produced by condensing, in the presence of a condensing agent, an aminated cyclodextrin, which is obtained by aminating one or more hydroxyl groups at 6-position, 3-position or 2-position of one or more sugar residues constituting the cyclodextrin, with galacturonic acid, to produce GalGCD represented by formula (1). For example, the above production method can produce GalGCD having a structure in which the amino group(s) of aminated GCD and the carboxy group at the 6-position carbon of galacturonic acid are bonded via an amide bond. It is sufficient if the aminated GCD has an amino group added to one or more of the carbon at 6-position, 3-position or 2-position of a sugar residue of GCD. GCD in which amino groups have been added to different sugar residues can be used, and GCD in which amino groups have been added to a plurality of carbon atoms within the same sugar residue can also be used. The number of amino groups is also not particularly limited, and GCD in which a plurality of amino groups have been added to one sugar residue or a plurality of sugar residues can be used. The formula (4) represents the GalGCD in which an aminated GCD obtained by substituting the hydroxyl group at 6-position of one sugar residue of GCD with an amino group and the carboxyl group at the 6-position carbon of galacturonic acid are bonded via an amide bond, and the formula (5) represents the GalGCD in which an aminated GCD obtained by substituting the hydroxyl group at 3-position of one sugar residue of GCD with an amino group and the carboxyl group at the 6-positon carbon of galacturonic acid are bonded via an amide bond.

[0045] The GalGCD of the present invention can be produced using a method known in the art, for example, by referring to Patent Literature 7 (JP 2020-37673 A). Specifically, it can be produced as follows. Aminated GCD can be prepared by tosylating GCD, azidating the resulting tosylated GCD, and aminating the resulting azidated GCD. For example, the hydroxyl group at 6-position of GCD can be converted to an amino group by tosylating the hydroxyl group with, for example, p-toluenesulfonyl chloride (tosyl chloride). The tosylated hydroxyl group can then be converted to an azido group with sodium amide, and finally the azido group can be reduced with triphenylphosphine to obtain aminated GCD. The tosylated hydroxyl group can also be converted to an amino group more simply by reacting it with aqueous ammonia to obtain aminated GCD, but other synthesis methods are also possible. Furthermore, aminated GCD can also be prepared by chlorinating GCD, azidating the resulting chlorinated GCD, and aminating the resulting azidated GCD. Alternatively, aminated GCD with a specific degree of substitution can be synthesized by after tosylation or chlorination, separating tosylated or chlorinated GCD with a specific degree of substitution by liquid chromatography, followed by azidation and amination. Furthermore, various aminated GCDs available commercially as reagents can also be purchased and used. Aminated GCD salts, in which the aminated GCD is in the form of a salt with an acid such as hydrochloric acid, may also be used.

[0046] The condensing agent used in the production of GalGCD of the present invention may be any agent capable of forming the amide bond, and any commonly used condensing agent may be used. Specifically, examples of condensing agents that can be used include 1H-benzotriazol-1-yloxytris(dimethylamino)phosphonium hexafluorophosphate (BOP reagent), 1-hydroxybenzotriazole (HOBt reagent), 1H-benzotriazol-1-yloxytripyrrolidinophosphonium hexafluorophosphate (PyBOP reagent), N,N-dicyclohexylcarbodiimide (DCC reagent), 1-[3-(dimethylamino)propyl]-3-ethylcarbodiimide (WSC reagent), N,N'-diisopropylcarbodiimide (DIC reagent), 4-(4,6-dimethoxy-1,3,5-triazin-2-yl)-4-methylmorpholinium chloride (DMT-MM reagent), and O-(benzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate (HBTU reagent).

[0047] The condensation reaction using a condensing agent in the production of GalGCD of the present invention may be appropriately adjusted depending on the properties of the condensing agent used, and, for example, in the case of the BOP reagent, the reaction may be carried out at room temperature in N,N-dimethylformamide (DMF) for 3 hours.

[0048] 2. Pharmaceutical Compositions The pharmaceutical composition of the present invention, which comprises GalGCD as an active ingredient, can be used as a therapeutic agent for lysosomal diseases that are lipid storage diseases. Examples of lysosomal diseases that are lipid storage diseases include cholesterol storage disease, sphingolipidosis, and lysosomal acid lipase deficiency. Cholesterol storage diseases are diseases accompanied by the accumulation of cholesterol, and examples thereof include Niemann-Pick Disease Type C, Wolman Disease, Cholesterol Ester Storage Disease (CESD), Tangier Disease, and Familial Hypercholesterolemia. The pharmaceutical composition of the present invention is particularly preferably used for Niemann-Pick Disease Type C. Examples of sphingolipidoses include GM1 gangliosidosis, GM2 gangliosidosis, Gaucher disease, Fabry disease, etc. The pharmaceutical composition of the present invention is preferably used for GM1 gangliosidosis and GM2 gangliosidosis.

[0049] The modified γ-cyclodextrin, which is the active ingredient of the pharmaceutical composition of the present invention, is the GalGCD, and the pharmaceutical composition of the present invention reduces the accumulation of lipid, particularly cholesterol, and / or suppresses disorder of the autophagy pathway.

[0050] "Reducing cholesterol accumulation" or "accumulation of cholesterol is reduced" means, for example, that when administered to NPC model cells (e.g., neural stem cells induced from iPS cells derived from an NPC patient) at a concentration of, for example, 1 mM, the accumulation of free cholesterol in the cells is reduced, preferably by 10% or more, 20% or more, 30% or more, 40% or more, 45% or more, or about 50% compared to when administration is not carried out. Alternatively, "reducing cholesterol accumulation" means, for example, when administered to NPC model cells at a concentration of, for example, 1 mM, the amount of esterified cholesterol (the ratio of esterified cholesterol to total cholesterol) in the cells is increased, preferably by 1.1-fold or more, 1.3-fold or more, 1.5-fold or more, 1.8-fold or more, 2-fold or more, 2.2-fold or more, 2.4-fold or more, or about 2.5-fold compared to when administration is not carried out. In one embodiment, the pharmaceutical composition of the present invention exhibits the effect of "reducing cholesterol accumulation" to the same extent as a pharmaceutical composition containing the same concentration of 2-hydroxypropyl-β-cyclodextrin (HPBCD) or 2-hydroxypropyl-γ-cyclodextrin (HPGCD).

[0051] "Suppressing disorder of the autophagy pathway" means that when administered to NPC model cells (e.g., neural stem cells induced from iPS cells derived from an NPC patient) at a concentration of, for example, 1 mM, the autophagosome volume is reduced, preferably by 5% or more, 10% or more, 15% or more, 20% or more, 25% or more, or about 30% compared to when administration is not carried out. In one aspect, the pharmaceutical composition of the present invention exhibits the function of "suppressing disorder of the autophagy pathway" to the same extent as a pharmaceutical composition containing the same concentration of 2-hydroxypropyl-β-cyclodextrin (HPBCD) or 2-hydroxypropyl-γ-cyclodextrin (HPGCD).

[0052] The GalGCD has lower toxicity than HPBCD, HPGCD, glucosyl-GCD, and maltosyl-GCD, in particular those for ototoxicity is problematic. Without limitation, "low ototoxicity" means, for example, that the effect on the hearing threshold due to administration is lower than that of HPBCD, HPGCD, glucosyl-GCD, and maltosyl-GCD, and preferably means that the sound pressure threshold is significantly lower over a wide range of frequencies (from low to high frequency).

[0053] The subjects to which the pharmaceutical composition of the present invention is administered include, for example, humans and non-human mammals (e.g., guinea pigs, rats, mice or other rodents, rabbits, cats, dogs, pigs, sheep, goats, cows, horses, donkeys, and non-human primates), and preferably humans.

[0054] The pharmaceutical composition of the present invention can be in the form of a parenteral administration form, preferably an injectable preparation, but is not limited thereto. The parenteral administration agent (e.g., an injectable preparation) of the present invention can be administered intravenously, intramuscularly, subcutaneously, etc., or can also be administered directly into the cerebral ventricles. The pharmaceutical composition of the present invention can be in the form of either an aqueous preparation or a lyophilized preparation, preferably an aqueous injection or a lyophilized injection to be reconstituted at the time of use.

[0055] The composition of the present invention may contain sugars, preservatives, stabilizers, and antistatic agents commonly used in injections. The composition of the present invention may also contain a pharmacologically acceptable pH adjuster. The pH adjuster used in the present invention is not particularly limited as long as it is a pharmacologically acceptable substance that can be used for pharmaceutical purposes, but is preferably sodium hydroxide, carbonate buffer, phosphate buffer, citrate buffer, acetate buffer, or hydrochloric acid. These pH adjusters may be used alone or in combination of two or more. The composition of the present invention may further contain an osmotic pressure adjuster or isotonicity agent, and may contain at least one selected from, for example, sodium chloride, dextrose and the like.

[0056] The effective dose of the pharmaceutical composition of the present invention can be appropriately selected depending on the type of disease, severity of the disease, treatment plan, administration route, body weight, age, sex, and (genetic) racial background of the patient. However, the pharmaceutically effective amount is generally determined based on factors such as clinically observed symptoms and the degree of progression of the disease. The route of administration is not particularly limited, and examples include intravenous infusion, intracerebroventricular infusion, intrathecal infusion, subarachnoid infusion, lumbar puncture injection, and nasal administration. Depending on the administration route, the daily dose is, for example, about 1 mg / kg to about 2 g / kg, preferably about 5 mg / kg to about 500 mg / kg. Lower doses are administered for intracerebroventricular infusion, intrathecal infusion, subarachnoid infusion, and lumbar puncture injection compared to intravenous infusion. The administration may be a single dose or a plurality of doses, or may be continuously administered over a period of time by infusion, etc., but is preferably administered over several hours or more, for example, several to about 10 hours, by infusion. The administration may be daily or intermittent, and can be selected appropriately depending on the condition of the subject, but intermittent administration is preferred. For example, in the case of intracerebroventricular injection, about 1 mg / kg to about 50 mg / kg, preferably about 10 mg / kg to about 30 mg / kg, can be administered weekly or every other week for several months to several years. The dosage can be adjusted appropriately, taking into account the condition of the subject (e.g., the onset of ototoxicity, etc.).

[0057] Furthermore, the pharmaceutical composition of the present invention has excellent safety and can be administered over a long period of time. In other words, the lysosomal diseases targeted by the pharmaceutical composition of the present invention are genetic diseases, and in many cases administration is required for the patient's lifetime. Since the pharmaceutical composition of the present invention has excellent safety, it is particularly suitable for such uses. The period during which the pharmaceutical product of the present invention can be administered is not particularly limited, but the pharmaceutical composition of the present invention can be administered over a long period of time, for example, for at least several weeks or more, preferably for several months or more, and more preferably for several years or more.

[0058] As used herein, the term "therapeutically effective amount" refers to an amount of GalGCD of the present invention that is effective to produce a desired therapeutic effect, e.g., reduction of cholesterol accumulation and / or suppression of disorder of the autophagy pathway, with a reasonable benefit-to-risk ratio. The therapeutically effective amount may vary depending on the route of administration used, as known to those skilled in the art. Furthermore, the therapeutically effective amount may be appropriately determined depending on the subject being treated, the severity of the condition, the route of administration, the frequency of administration, the judgment of the prescribing physician, and other relevant factors.

[0059] As detailed in detail below, in neural stem cells induced from iPS cells derived from NPC patients, free cholesterol accumulates intracellularly, similar to that observed in patients. When GalGCD, which is the active ingredient of the pharmaceutical composition of the present invention, was examined for its effect on this accumulation, it demonstrated a similar reduction effect to that of HPGCD used as a control. Furthermore, GalGCD exhibited extremely low levels of toxicity to hearing, even after repeated intracerebroventricular administration. Furthermore, while abnormalities in autophagy function were observed in neural stem cells derived from NPC patients, GalGCD treatment improved this abnormal phenotype, confirming its effectiveness on functional abnormalities of neural cells.

[0060] The present invention will be specifically described below with reference to examples, but the present invention is not limited to the following examples. [Example 1] Synthesis of mono-6-O-p-toluenesulfonyl-γ-cyclodextrin 50 g of γ-cyclodextrin was dissolved in 500 mL of 4-methylpyridine and cooled on ice. 11 g of p-toluenesulfonyl chloride was added to the solution and allowed to react at room temperature for 15 hours. The reaction solution was added to 3.5 L of acetone, and the resulting precipitate was collected and washed with acetone. The collected precipitate was dissolved in 200 mL of boiling water, cooled to 4°C, and recrystallized. The recrystallization treatment was performed a total of three times, and then dried under reduced pressure to obtain 11.3 g of a white powder. FT-IR and NMR confirmed that the white powder was mono-6-O-p-toluenesulfonyl-γ-cyclodextrin.

[0061] [Example 2] Synthesis of 6-amino-6-deoxy-γ-cyclodextrin 9 g of mono-6-O-p-toluenesulfonyl-γ-cyclodextrin prepared in Example 1 was dissolved in 200 mL of 28% aqueous ammonia and reacted at 60°C for 16 hours. The reaction solution was added to 2.5 L of acetone, and the resulting precipitate was collected and washed with acetone. The collected precipitate was dried under reduced pressure to obtain 7.1 g of a yellow-white powder. FT-IR and NMR confirmed that the yellow-white powder was 6-amino-6-deoxy-γ-cyclodextrin.

[0062] [Example 3] Synthesis of Gal6GCD (γ-cyclodextrin having a monovalent group derived from galacturonic acid at the 6-position carbon) 3 g of 6-amino-6-deoxy-γ-cyclodextrin prepared in Example 2 was dissolved in 30 mL of dimethylformamide (DMF), and 1.3 g of 1H-benzotriazol-1-yloxytris(dimethylamino)phosphonium hexafluorophosphate (BOP reagent) and 1.5 mL of N,N-diisopropylethylamine (DIEA) were added. 0.64 g of galacturonic acid monohydrate was added to the solution, and the mixture was sealed with Ar gas and reacted at ambient temperature for 3 hours. The reaction solution was added to 500 mL of acetone, and the resulting precipitate was collected and washed with acetone and methanol. The collected precipitate was dried under reduced pressure and then fractionated by preparative high-performance liquid chromatography using an ODS column. The fractionated solution was lyophilized, and 0.97 g of a white powder was collected. NMR confirmed that the white powder was Gal6GCD (compound represented by formula (4) below).

[0063]

[0064] [Example 4] Synthesis of 3A-amino-3A-deoxy-(2AS,3AS)-γ-cyclodextrin 7.6 g of mono-2-O-p-toluenesulfonyl-γ-cyclodextrin was dissolved in 300 mL of 10% (w / w) aqueous ammonium bicarbonate solution and reacted at 60°C for 5 hours. The reaction solution was added to 4 L of acetone, and the resulting precipitate was collected and washed with acetone. The collected precipitate was dried under reduced pressure to obtain a white powder. 6.6 g of the resulting white powder was dissolved in 100 mL of 28% aqueous ammonia and reacted at 60°C for 5 hours. The reaction solution was added to 1.5 L of acetone, and the resulting precipitate was collected and washed with acetone. The collected precipitate was dried under reduced pressure to obtain 7.1 g of a white powder. The precipitate was dissolved in 300 mL of 10% (w / w) aqueous ammonium bicarbonate solution and reacted at 60°C for 5 hours. The reaction solution was added to 4 L of acetone, and the resulting precipitate was collected and washed with acetone. The collected precipitate was dried under reduced pressure to obtain 5.4 g of a yellow-white powder. The yellow-white powder was identified as 3A-amino-3A-deoxy-(2AS,3AS)-γ-cyclodextrin by FT-IR and NMR.

[0065] [Example 5] Synthesis of Gal3GCD (γ-cyclodextrin having a monovalent group derived from galacturonic acid at the 3-position carbon) 2 g of 3A-amino-3A-deoxy-(2AS,3AS)-γ-cyclodextrin prepared in Example 4 was dissolved in 25 mL of DMF, and 0.82 g of BOP reagent and 0.97 mL of DIEA were added. 0.39 g of galacturonic acid monohydrate was added to the solution, and the mixture was sealed with Ar gas and reacted at ambient temperature for 3 hours. The reaction solution was added to 500 mL of acetone, and the resulting precipitate was collected and washed with acetone and methanol. The collected precipitate was dried under reduced pressure and then fractionated by preparative high-performance liquid chromatography using an ODS column. The fractionated solution was lyophilized, and 1.8 g of a white powder was recovered. NMR confirmed that the white powder was Gal3GCD (compound represented by formula (5) below).

[0066]

[0067] [Comparative Example 1] Synthesis of Glc6GCD (γ-cyclodextrin having a monovalent group derived from glucuronic acid at the 6-position carbon) 3.6 g of 6-amino-6-deoxy-γ-cyclodextrin prepared in Example 2 was dissolved in 30 mL of DMF, and 1.8 g of BOP reagent and 2.2 mL of DIEA were added. 0.81 g of glucuronic acid was added to the solution, and the mixture was sealed with Ar gas and reacted at ambient temperature for 3 hours. The reaction solution was added to 500 mL of acetone, and the resulting precipitate was collected and washed with acetone and methanol. The collected precipitate was dried under reduced pressure and then fractionated by preparative high-performance liquid chromatography using an ODS column. The fractionated solution was lyophilized, and 0.78 g of a white powder was recovered. NMR confirmed that the white powder was Glc6GCD (compound of formula (6) below).

[0068]

[0069] [Comparative Example 2] Synthesis of Glc3GCD (γ-cyclodextrin having a monovalent group derived from glucuronic acid at the 3-position carbon) 3.0 g of 3A-amino-3A-deoxy-(2AS,3AS)-γ-cyclodextrin prepared in Example 4 was dissolved in 25 mL of DMF, and 1.2 g of BOP reagent and 1.5 mL of DIEA were added. 0.54 g of glucuronic acid was added to the solution, and the mixture was sealed with Ar gas and reacted at ambient temperature for 3 hours. The reaction solution was added to 500 mL of acetone, and the resulting precipitate was collected and washed with acetone and methanol. The collected precipitate was dried under reduced pressure and then fractionated by preparative high-performance liquid chromatography using an ODS column. The fractionated solution was lyophilized, and 1.1 g of a white powder was recovered. NMR confirmed that the white powder was Glc3GCD (compound represented by formula (7) below).

[0070]

[0071] [Example 6] Effect of GalGCD on NPC model cells As NPC model cells, Npc1-deficient CHO cells (Npc1 null CHO cells: Higaki, K. et al., J. Biochem. 129, 875-880. (2001)) were used. After pre-culturing the NPC model cells for 24 hours, Gal6GCD and Gal3GCD were added to the medium at concentrations ranging from 0.001 to 10 mM. After 24 hours, the total cholesterol (TC; nmol) and free cholesterol (FC; nmol) levels per mg of protein in the cells were measured. Wild-type (WT) CHO cells were used as controls (no CDs added). Glc6GCD, Glc3GCD, hydroxypropyl-β-cyclodextrin (HP-β-CD), and hydroxypropyl-γ-cyclodextrin (HP-γ-CD) were used as reference compounds. Specifically, the NPC model cells were treated with or without each γ-cyclodextrin derivative for 24 hours and then lysed. Protein concentrations in the lysates were measured using a protein assay kit. The protein concentration results are shown in Figure 1. Addition of γ-cyclodextrin derivatives at concentrations up to 10 mM did not significantly decrease total protein levels. Cholesterol was then extracted from the lysates and the cholesterol level was measured using standard methods. A portion of the extract was incubated together with esterase to measure total cholesterol, while another portion was incubated without esterase to measure free cholesterol. Cholesterol levels were calculated in nmol / mg protein. Furthermore, cholesterol levels in Npc1-deficient CHO cells treated with γ-cyclodextrin derivatives are shown as cholesterol accumulation rates, relative to the intracellular cholesterol content of wild-type cells (set at 100). The results of total cholesterol measurements are shown in Figure 2, and the results of free cholesterol are shown in Figure 3. The ratio of esterified cholesterol to total cholesterol is shown in Figure 4. The addition of GCD did not affect cellular protein levels.

[0072] In the NPC model cells, both GlcGCD and GalGCD concentration-dependently significantly reduced total cholesterol, reduced free cholesterol, and increased the esterified cholesterol ratio, demonstrating a significant ameliorative effect on intracellular cholesterol accumulation. Furthermore, both CDs with Glc or Gal attached to the 6-position carbon of GCD and CDs with Glc or Gal attached to the 3-position carbon of GCD showed significant ameliorative effects. Specifically, administration of GlcGCD and GalGCD (1 mM) reduced intracellular free cholesterol accumulation by approximately 50% compared to non-administered cases, and increased intracellular esterified cholesterol by more than two-fold compared to non-administered cases. The ameliorative effect on intracellular cholesterol accumulation of both GlcGCD and GalGCD was comparable to that of HP-β-CD and HP-γ-CD. Furthermore, it was confirmed that even when a GalGCD concentration 20-100 times higher (10 mM) than the effective concentration (0.1-0.5 mM) at which GalGCD exhibits an improving effect was used, no significant decrease in total protein amount was observed, so it can be assumed that cytotoxicity is low.

[0073] [Example 7] Effect of GalGCD on neural stem cells induced from NPC-iPS cell line An NPC-iPS cell line prepared from a patient with an NPC1 mutation was used. Japanese republished application 2015 / 083736 discloses the production of NPC-iPS cell lines and is incorporated herein by reference in its entirely. Neural stem cells induced from iPSCs derived from NPC patients exhibit intracellular accumulation of free cholesterol, similar to that observed in patients. A normal cell line (iPS cell line produced from normal fibroblasts) and an NPC-iPS cell line were induced to differentiate into neural stem cells using known methods. After pre-culturing each of the differentiated cells, Gal6GCD and Gal3GCD were added to the medium at concentrations of 100 μM, 300 μM, and 1 mM. After 96 hours, Filipin staining was performed and then analyzed using an IN CELL ANALYZER (GE Healthcare). Filipin staining can detect intracellular free cholesterol and confirm cholesterol accumulation. HPGCD was used as a reference compound. To confirm the presence of cells, nuclei were stained with propidium iodide and Hoechst 33258. The results of nuclear staining and Filipin staining observed under a fluorescence microscope are shown in Figure 5A, and a graph of the degree of decrease in Filipin staining fluorescence is shown in Figure 5B.

[0074] Compared to normal cell lines, NPC-iPS cell lines were confirmed to have significant intracellular cholesterol accumulation, but when Gal3GCD, Gal6GCD, and HPGCD were added, a concentration-dependent decrease in intracellular cholesterol accumulation was observed.

[0075] [Example 8] Evaluation of the effect of GalGCD on the recovery of autophagy function In patients with this disease, the expression of LC3B-II protein, which is a marker for autophagy initiation, is elevated, indicating that autophagy is activated, but the accumulation of insoluble p62 protein suggests that autophagy does not progress and is stalled. Similarly, abnormalities in the autophagy function of cells were observed in neural stem cells differentiated from NPC-iPS cell lines prepared from patients with NPC1 mutations. Using neural stem cells induced from NPC-iPS cell lines, we measured the effect of GalGCD on reducing LC3B-II protein expression (A) and insoluble p62 (B), and evaluated the effect of GalGCD on recovering autophagy function. Gal3GCD, Gal6GCD, and HPGCD were added at a concentration of 1 mM, and LC3B-II protein and insoluble p62 protein were measured 72 hours later. The results are shown in Figure 6. From the above, it was found that abnormalities in autophagy function were observed in neural stem cells derived from NPCs, but this abnormal phenotype was improved by GalGCD treatment.

[0076] [Example 9] Evaluation of ototoxicity of GalGCD To evaluate ototoxicity, Gal3GCD and Gal6GCD were repeatedly administered intracerebroventricularly to 9-week-old normal adult mice, and the auditory brainstem response (ABR) was measured. Administration was performed every two weeks at a dose of 0.029 nmol / kg, for a total of five doses, and ABR measurements were performed three days after the final administration. ABR measurements were performed in each animal at four frequencies (4, 8, 12, and 20 kHz). HPGCD was used as the reference compound and administered in the same manner. The results are shown in Figure 7.

[0077] Non-Patent Literature 4 reports that g-cyclodextrin derivatives having glucosyl groups attached thereto retain ototoxicity equivalent to that of HPGCD when administered intracerebroventricularly. On the other hand, it was found that both Gal3GCD and Gal6GCD of the present invention have significantly lower ototoxicity than HPGCD. Accordingly, it was found that the two compounds Gal3GCD and Gal6GCD have cholesterol accumulation-reducing effect and neurological function-improving effect equivalent to HPGCD, and exhibit weaker side effects of ototoxicity than HPGCD.

[0078] The forgoing detailed description is merely intended to illustrate the objects and scope of the present invention, and is not intended to limit the scope of the appended claims. Various modifications and substitutions to the described embodiments will be apparent to those skilled in the art from the teachings set forth herein without departing from the scope of the appended claims.

[0079] The present invention provides a pharmaceutical composition that can be used for the treatment or prevention of lysosomal diseases, particularly Niemann-Pick disease. The cyclodextrin derivative, which is the active ingredient of the pharmaceutical composition of the present invention, is useful because it has cholesterol accumulation-reducing effect and neurological function-improving effect equivalent to conventional cyclodextrins, and has low ototoxicity.

Claims

1. A pharmaceutical composition for treating or preventing a lysosomal disease, comprising a modified γ-cyclodextrin as an active ingredient, wherein, in the modified γ-cyclodextrin, a monovalent group derived from galacturonic acid is bonded to a sugar residue of the γ-cyclodextrin via an amide bond.

2. The pharmaceutical composition according to claim 1, wherein, in the modified γ-cyclodextrin, a monovalent group derived from galacturonic acid is bonded to the 6-position carbon and / or the 3-position carbon of a sugar residue of the γ-cyclodextrin via an amide bond.

3. The pharmaceutical composition according to claim 1, wherein the modified γ-cyclodextrin is a compound represented by the following formula (1): wherein, m is 0 to 7, n is 0 to 8, o is 0 to 8, and m+n+o=8, R1and R2groups each independently represent a hydroxyl group or the formula (a): -NH-Z (a) wherein Z represents a monovalent group formed by removing a hydroxy group (-OH) from the carboxy group (-COOH) of galacturonic acid, provided that R1 and R2 are not both hydroxyl groups, and R3, R4and R5groups each independently represent a hydroxy group or the formula (a), optionally o R3and R4groups each independently represent a hydroxyl group or the formula (a), and o R5group represents the formula (a), wherein, in the formula (1), when n+o is 2 or more, the arrangement of structures (A), (B), and (C) below is arbitrary, and each (B) may be the same or different, and each (C) may be the same or different. .

4. The pharmaceutical composition according to claim 1, wherein, the modified γ-cyclodextrin is a compound represented by the following formula (2): wherein, m is 0 to 7, n is 1 to 8, and m+n=8, and R1and R2groups each independently represent a hydroxyl group or the formula (a): -NH-Z (a) wherein Z represents a monovalent group formed by removing the hydroxy group (-OH) from the carboxy group (-COOH) of galacturonic acid, wherein, in formula (2), when n is 2 or more, the arrangement of structure (A) and (B) below is arbitrary, and each (B) may be the same or different, or by the following formula (3): wherein, m is 0 to 7, o is 1 to 8, and m+o=8, and R3, R4and R5groups each independently represent a hydroxyl group or the formula (a): -NH-Z (a) wherein, Z represents a monovalent group formed by removing the hydroxy group (-OH) from the carboxy group (-COOH) of galacturonic acid, wherein, in formula (3), when o is 2 or more, the arrangement of structure (A) and (C) below is arbitrary, and each (C) may be the same or different, .

5. The pharmaceutical composition according to claim 4, wherein the modified γ-cyclodextrin is represented by the formula (2), wherein n is 1, R1is the formula (a), and R2is OH.

6. The pharmaceutical composition according to claim 4, wherein the modified γ-cyclodextrin is represented by the formula (3), wherein o is 1, R3and R4are OH, and R5is the formula (a).

7. The pharmaceutical composition according to any one of claims 1 to 6, wherein the lysosomal disease is a lipid storage disease.

8. The pharmaceutical composition according to claim 7, wherein the lipid storage disease is selected from the group consisting of Niemann-Pick disease type C, GM1 gangliosidosis, and GM2 gangliosidosis.

9. The pharmaceutical composition according to any one of claims 1 to 6, wherein the pharmaceutical composition is administered parenterally.

10. The pharmaceutical composition according to any one of claims 1 to 6, wherein the pharmaceutical composition is administered as an injection.

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