Treatment agent for cancer cachexia or sarcopenia, containing nanoparticles derived from polymerized cyclic nitroxide radical compound as active ingredient

Abstract

Disclosed in the present invention is a cancer cachexia prophylactic or therapeutic agent which is a formulation containing, as an active ingredient, nano-sized particles consisting essentially of a graft polymer, or a base-added salt thereof, in which a residue of a cyclic nitroxide radical compound, a residue of a trialkoxyalkylsilane, and a poly(ethylene glycol) chain are inserted into a side chain of a poly(styrene-maleic anhydride) copolymer constituting the basic skeleton. When orally ingested, the prophylactic or therapeutic agent remains in the gastrointestinal tract for a long period of time with virtually no transfer into the blood and can reduce the symptoms of cancer cachexia or sarcopenia. Thus, the present disclosure provides a pharmaceutical formulation capable of effectively preventing or treating cancer cachexia or sarcopenia.

Description

A treatment for cancer cachexia or sarcopenia containing nanoparticles derived from polymerized cyclic nitroxide radical compounds as an active ingredient

[0001] The present invention relates to an agent for treating (preventing or treating) cancer cachexia or sarcopenia, which contains, as an active ingredient, nanoparticles derived from a polymerized cyclic nitroxide radical compound or a base addition salt thereof. More specifically, the present invention relates to an agent for preventing or treating cancer cachexia or sarcopenia, which contains, as an active ingredient, nanoparticles derived from an amphiphilic graft polymer having a poly(styrene-maleic anhydride) copolymer (PSMA) as a basic skeleton and having cyclic nitroxide radicals, poly(ethylene glycol) chains, and alkoxysilane groups introduced into its side chains.

[0002] The present inventors have discovered that block copolymers containing a segment having a repeating unit in which a cyclic nitroxide radical compound is introduced into the side chain and a poly(ethylene glycol) (PEG) segment, or nanoparticles formed from such copolymers in an aqueous medium, can be useful for preventing or treating diseases caused by the production or presence of excessive reactive oxygen species (ROS) in the body by utilizing the redox function that the nitroxide radical can exert in a biological environment (see, for example, WO 2009 / 133647 A1). Furthermore, the present inventors have provided an amphiphilic graft polymer in which alkoxysilane groups for forming chemical crosslinks between the cyclic nitroxide radical compound and the poly(ethylene glycol) chain are introduced into poly(styrene-co-maleic anhydride) copolymer (hereinafter sometimes abbreviated as PSMA) via the maleic anhydride repeating unit, in order to provide nanoparticles containing residues derived from a polymerized cyclic nitroxide radical compound (see, for example, JP 2019-123773 A). The present inventors have clarified that the graft polymer can provide nano-sized particles (so-called core-shell micelles, hereinafter also referred to as nanoparticles, which contain the site associated with the chemical crosslinking as a main core and a shell mainly composed of PEG chains) that are stably retained in an aqueous medium and exhibit dispersion stability.

[0003] Furthermore, the present inventors have developed nanoparticles or nano-sized particles (hereinafter referred to as siSMAPo) formed from the graft polymer. ＴＮ ) is a high molecular weight antioxidant that distributes only in the intestinal tract after oral administration and does not diffuse throughout the body. It has been discovered and published that it protects the intestinal environment by selectively eliminating reactive oxygen species in the intestines of experimental animals damaged by chronic restraint stress (hereinafter sometimes abbreviated as CRS), and furthermore, it suppresses physiological and behavioral depression-related symptoms in CRS (see Yutaka IKEDA, Naoki SAIGO, Yukio NAGASAKI, Biomaterials 295 (2023) 122053). The authors of this Biomaterials (including the present inventor) are the same as those of the aforementioned siSMAPo ＴＮ In light of the characteristics of siSMAPo ＴＮ We believe that this compound selectively scavenges excess reactive oxygen species in the intestine and is a novel antidepressant candidate that can be used to treat depression mediated by the gut-brain axis.

[0004] Meanwhile, numerous studies have reported that the gut interacts with various organs and influences their function in relation to the gut-brain axis. These interactions are variously referred to as the gut-liver axis, gut-lung axis, and other terms, including the gut-brain axis, but in all cases, the concept is that the intestinal environment interacts with distant organs via the autonomic nervous system, hormones, cytokines, and intestinal bacterial metabolites. Based on this concept, it is believed that controlling the intestinal environment can also control the function of various organs. This strategy is unique in that it does not directly target the disease site, but targets the intestine, improving the intestinal environment and utilizing the body's inherent immune system homeostasis to treat disease. In fact, probiotics, prebiotics, and fecal transplants have been attempted to treat various diseases. However, these attempts have had limited effectiveness, and fecal transplants have also caused severe side effects.

[0005] As discussed therein, the above-mentioned Biomaterials is, to the best of the inventors' knowledge, the first report to reveal that CRS-induced depressive symptoms can be prevented through the elimination of intestinal reactive oxygen species. Based on the concept that the intestinal environment interacts with distant organs in the body via the autonomic nervous system, hormones, cytokines, intestinal bacterial metabolites, etc., as mentioned above, if it is possible to approach the prevention or treatment of diseases or injuries other than those mediated by the gut-brain axis, then the above-mentioned siSMAPo could be used for various diseases for which probiotics, prebiotics, fecal transplants, etc. have been attempted to control the intestinal environment. ＴＮ may be effective.

[0006] From the above viewpoint, siSMAPo ＴＮ Cancer cachexia was selected as a disease or injury for which the agent may be effective, because its onset mechanism and test animal model are fundamentally different from CRS. Therefore, a primary object of the present invention is to provide a new, more effective agent or method for treating (preventive or therapeutic) cancer cachexia.

[0007] Cancer cachexia is one of the complications seen in cancer patients and is considered synonymous with malnutrition accompanied by cancer-related inflammation. Its main symptoms are loss of appetite, weight loss, and loss of muscle mass, and it is observed in 50% to 80% of cancer patients. Currently, there is one drug (anamorelin hydrochloride) that targets cancer cachexia, but although appetite has been improved, no improvement in muscle strength has been confirmed.

[0008] Therefore, instead of using mice damaged by CRS as a model experimental animal, the present inventors used a cancer cachexia model created by subcutaneously transplanting mouse colon cancer cells, and ＴＮ The siSMAPo-containing aqueous solution was administered ad libitum. ＴＮ The effects of administration were examined.

[0009] As a result, the siSMAPo ＴＮIt was confirmed that the treatment group showed the following improvements compared to the untreated group: (a) a significant suppression of the decrease in quadriceps muscle mass, a typical skeletal muscle, and muscle mass was maintained, which are the main symptoms or biological parameters associated with cancer cachexia; (b) a significant improvement in muscle strength; and (c) a significant suppression of the increase in the expression level of atrogin-1, a type of ubiquitin ligase that plays a central role in muscle protein degradation.

[0010] Then, siSMAPo ＴＮ It can be said that this study has specifically confirmed that siSMAPo significantly improves the main symptoms of cancer cachexia, which is a disease or injury that is different from depression mediated by the gut-brain axis. ＴＮ Based on the aforementioned concept that intestinal bacteria interact with each other via intestinal bacterial metabolites, it has been confirmed that it is possible to control the functions of various organs by controlling the intestinal environment. Therefore, for example, siSMAPo can be used for the prevention or treatment of sarcopenia, which is caused by an increase in systemic inflammation associated with aging, a decrease in muscle mass, and a decline in muscle strength and physical function. ＴＮ is presumed to be valid.

[0011] siSMAPo ＴＮ is not limited to those prepared by particle formation treatment from a graft polymer: siPSMS(T)-g-PEG (with structural formula) produced according to the <Reaction Scheme> described below (Production Example 1: Production of Graft Polymer and Preparation of Nanoparticles from the Graft Polymer), and the object of the present invention can be achieved even if the graft polymer is nano-sized particles prepared from a graft polymer represented by Formula I or a base addition salt thereof, which is generally defined in Aspect 1 of the present application, as long as they are nano-sized particles that exhibit dispersion stability in an aqueous medium and can control the intestinal environment by, for example, scavenging excess intestinal reactive oxygen species. For example, the graft polymer represented by Formula I is amphiphilic and can disperse the siSMAPo ＴＮThis is because it is understood that nano-sized particles substantially equivalent to

[0012] In light of the above background, the subject matter or invention disclosed in this specification mainly has the following features or aspects.

[0013] Aspect 1: A preventive or therapeutic agent for cancer cachexia or sarcopenia, comprising, as an active ingredient, nano-sized particles essentially consisting of a graft polymer represented by the following formula I or a base addition salt thereof. Formula I: In the formula, a+b+c+d is an integer from 8 to 1400, n is an integer from 8 to 1400, a+b+c+d:n is in a ratio of 1:1 to 5, a+b+d:c is in a ratio of 1 to 60:1, a+b+c+d is a set of repeating units that can be represented as a whole by m, the repeating units of m and repeating units with the subscript n are mutually included randomly or alternately, and in m, repeating units with the symbols a, b, c, and d are each included randomly, with the proviso that repeating units with the symbol d are zero (or not present), or, if present, can account for up to 15% of the total number of a+b+c+d.

[0014] (1) In the repeating unit marked with the subscript a, L １ -TEMPO, L １ is NH or O, and TEMPO is a residue of a cyclic nitroxide radical compound selected from the group consisting of 2,2,6,6-tetramethylpiperidin-1-oxyl-4-yl, 2,2,5,5-tetramethylpyrrolidin-1-oxyl-3-yl, 2,2,5,5-tetramethylpyrrolin-1-oxyl-3-yl, 2,4,4-trimethyl-1,3-oxazolidine-3-oxyl-2-yl, 2,4,4-trimethyl-1,3-thiazolidine-3-oxyl-2-yl, and 2,4,4-trimethyl-imidazolindin-3-oxyl-2-yl.

[0015] (2) In the repeating unit marked with subscript b, R ｓｉ is expressed as follows: where R ３ is C １ -C３ where r is an integer of 1 to 6, and H in NH is C １ -C ６ It may be substituted by an alkyl group of the formula:

[0016] (3) In the repeating unit marked with subscript c, L ２ -PEG-A, L ２ is O or NR, where R is a hydrogen atom or C １ -C ６ and PEG is an alkyl group represented by the formula: where p is an integer of 1 to 6, q is an integer of 5 to 500, and A is: A1: unsubstituted or substituted C １ -C １２ represents an alkoxy group, and when substituted, the substituent is a formyl group, a group represented by the formula R ａ R ｂ CH- (where R ａ and R ｂ is independent, C １ -C ４ Alkoxy or R １ and R ２ Let's get together -OCH ２ CH ２ O-, -O(CH ２ ) ３ O- or -O(CH ２ ) ４ A2: a group of the following formula: The repeating units marked with c account for 2% to 15% of the total units of the graft polymer represented by formula I.

[0017] (4) In the repeating unit marked with subscript d, R １ and R ２ together represent —O— to form a cyclic anhydride residue, or each represent OH.

[0018] Aspect 2: The preventive or therapeutic agent for cancer cachexia or sarcopenia according to Aspect 1, wherein the preventive or therapeutic agent is orally administered.

[0019] Aspect 3: The preventive or therapeutic agent for cancer cachexia or sarcopenia according to Aspect 1, wherein the nano-sized particles consist of a graft polymer represented by Formula I or a base addition salt thereof.

[0020] Aspect 4: The agent for preventing or treating cancer cachexia or sarcopenia according to aspect 1, wherein the nano-sized particles are R in the repeating unit marked with the subscript b ｓｉ and a tetraalkoxysilane (wherein the alkoxy is C) for forming a chemical crosslink with １ -C ４ The preventive or therapeutic agent as described above, comprising a component derived from a compound selected from the group consisting of hydroxypropyl methyl, ...

[0021] Aspect 5: The agent for preventing or treating cancer cachexia or sarcopenia according to aspect 1, wherein the nano-sized particles are R in a repeating unit marked with a subscript b. ｓｉ or R ｓｉ The preventive or therapeutic agent as described above, comprising nano-sized silica adsorbed via

[0022] Aspect 6: The agent for preventing or treating cancer cachexia or sarcopenia according to Aspect 1, wherein TEMPO in the repeating unit with the subscript a is represented by the following formula: wherein R' is a methyl group.

[0023] Aspect 7: Nano-sized particles consisting essentially of a graft polymer represented by the following formula I or a base addition salt thereof for use in the prevention or treatment of cancer cachexia or sarcopenia: Formula I: In the formula, a+b+c+d is an integer from 8 to 1400, n is an integer from 8 to 1400, a+b+c+d:n is in a ratio of 1:1 to 5, a+b+d:c is in a ratio of 1 to 60:1, a+b+c+d is a group of repeating units that can be represented as a whole by m, the repeating units of m and repeating units with the subscript n are mutually randomly included, and in m, repeating units with the symbols a, b, c and d are each randomly included, with the proviso that the repeating units with the symbol d are zero (or not present), or, if present, can account for up to 15% of the total number of a+b+c+d.

[0024] (1) In the repeating unit marked with the subscript a, L １ -TEMPO, L １ is NH or O, and TEMPO is a residue of a cyclic nitroxide radical compound selected from the group consisting of 2,2,6,6-tetramethylpiperidin-1-oxyl-4-yl, 2,2,5,5-tetramethylpyrrolidin-1-oxyl-3-yl, 2,2,5,5-tetramethylpyrrolin-1-oxyl-3-yl, 2,4,4-trimethyl-1,3-oxazolidine-3-oxyl-2-yl, 2,4,4-trimethyl-1,3-thiazolidine-3-oxyl-2-yl, and 2,4,4-trimethyl-imidazolindin-3-oxyl-2-yl. (

[0025] (2) In the repeating unit marked with subscript b, R ｓｉ is expressed as follows: where R ３ is C １ -C ３ where r is an integer of 1 to 6, and H in NH is C １ -C ６ It may be substituted by an alkyl group of the formula:

[0026] (3) In the repeating unit marked with subscript c, L ２ -PEG-A, L ２ is O or NR, where R is a hydrogen atom or C １ -C ６ and PEG is an alkyl group represented by the formula: where p is an integer of 1 to 6, q is an integer of 5 to 500, and A is A1: unsubstituted or substituted C １ -C １２ represents an alkoxy group, and when substituted, the substituent is a formyl group, a group represented by the formula R ａ R ｂ CH- (where R ａ and R ｂ is independent, C １ -C ４ Alkoxy or R １ and R ２ Let's get together -OCH ２ CH ２ O-, -O(CH ２ ) ３ O- or -O(CH ２ ) ４ A2: a group of the following formula: and the repeating units account for 2% to 15% of the total units of the graft polymer represented by formula I.

[0027] (4) In the repeating unit marked with subscript d, R １ and R ２ together represent —O— to form a cyclic anhydride residue, or each represent OH.

[0028] Aspect 8: A method for preventing or treating cancer cachexia or sarcopenia, comprising the step of administering an effective amount of nano-sized particles consisting essentially of a graft polymer represented by the following formula I or a base addition salt thereof to a patient suffering from or likely to suffer from cancer cachexia or sarcopenia: Formula I: In the formula, a+b+c+d is an integer from 8 to 1400, n is an integer from 8 to 1400, a+b+c+d:n is in a ratio of 1:1 to 5, a+b+d:c is in a ratio of 1 to 60:1, a+b+c+d is a group of repeating units that can be represented as a whole by m, the repeating units of m and repeating units with the subscript n are mutually randomly included, and in m, repeating units with the symbols a, b, c and d are each randomly included, with the proviso that the repeating units with the symbol d are zero (or not present), or, if present, can account for up to 15% of the total number of a+b+c+d.

[0029] (1) In the repeating unit marked with the subscript a, L １ -TEMPO, L １ is NH or O, and TEMPO is a residue of a cyclic nitroxide radical compound selected from the group consisting of 2,2,6,6-tetramethylpiperidin-1-oxyl-4-yl, 2,2,5,5-tetramethylpyrrolidin-1-oxyl-3-yl, 2,2,5,5-tetramethylpyrrolin-1-oxyl-3-yl, 2,4,4-trimethyl-1,3-oxazolidine-3-oxyl-2-yl, 2,4,4-trimethyl-1,3-thiazolidine-3-oxyl-2-yl, and 2,4,4-trimethyl-imidazolindin-3-oxyl-2-yl.

[0030] (2) In the repeating unit marked with subscript b, R ｓｉ is expressed as follows: where R ３ is C １ -C ３ where r is an integer of 1 to 6, and H in NH is C １ -C ６ It may be substituted by an alkyl group of the formula:

[0031] (3) In the repeating unit marked with subscript c, L ２ -PEG-A, L ２ is O or NR, where R is a hydrogen atom or C １ -C ６ and PEG is an alkyl group represented by the formula: where p is an integer of 1 to 6, q is an integer of 5 to 500, and A is: A1: unsubstituted or substituted C １ -C １２ represents an alkoxy group, and when substituted, the substituent is a formyl group, a group represented by the formula R ａ R ｂ CH- (where R ａ and R ｂ is independent, C １ -C ４ Alkoxy or R １ and R ２ Let's get together -OCH ２ CH２ O-, -O(CH ２ ) ３ O- or -O(CH ２ ) ４ A2: a group of the following formula: and the repeating units account for 2% to 15% of the total units of the graft polymer represented by formula I.

[0032] (4) In the repeating unit marked with subscript d, R １ and R ２ together represent —O— to form a cyclic anhydride residue, or each represent OH.

[0033] According to the disclosure of this specification, a use invention is disclosed for using nano-sized particles essentially consisting of the graft polymer represented by the formula I in the prevention or treatment of cancer cachexia or sarcopenia. The effects of this use invention are based on the fact that the particles are suitable for oral administration, stably remain in the intestinal tract, exert redox function, and selectively eliminate excess reactive oxygen species in the intestine, thereby effectively suppressing disturbances in the intestinal environment.

[0034] 1 is a conceptual diagram of nanoparticles formed by self-assembly from the copolymer disclosed herein. 2 is a graphical representation of body weight change in Test Example 1 (1). 3 is a graphical representation of tumor volume change in Test Example 1 (2). 4 is a graphical representation of the results of muscle strength measurement in Test Example 1 (3). 5 is a graphical representation of the results of blood IL-6 level measurement in Test Example 2 (1). 6 is a graphical representation of the results of quadriceps muscle weight measurement in Test Example 2 (2). 7 is a graphical representation of the results of Western blotting regarding Atrogin 1 expression levels in Test Example 2 (3) and the analytical results thereof (b). Detailed Description of the Invention

[0035] Nanoparticles essentially consisting of the graft polymer represented by Formula I or a base addition salt thereof are generally described in the above-mentioned Japanese Patent Publication No. 2019-123773, and specific and typical examples can be prepared according to the description in Biomaterials 295(2023)122053 (hereinafter sometimes abbreviated as Biomaterials'053) and in Preparation Example 1 described later in this specification. Therefore, the specific and typical nanoparticles disclosed in this specification are understood to have substantially the same physicochemical characteristics in light of both the above descriptions. This publication and Biomaterials'053 are hereby incorporated by reference, and all matters described therein are incorporated by reference herein.

[0036] Terms used herein, particularly technical terms, have the meanings commonly used in the art unless otherwise defined.

[0037] The terms "comprise" and "comprising" mean that any components other than the listed components may be included. The terms "essentially consisting of" and "consist essentially of" mean that the listed components are the most important components and may further include other components as long as the intended purpose of the resulting product is not adversely affected. For example, in the present invention, when referring to nano-sized particles essentially consisting of a graft polymer represented by Formula I or a base addition salt thereof, other components than the graft polymer or a base addition salt thereof may be included as long as the other components do not adversely affect or alter the properties of the nano-sized particles. Examples of such components include, but are not limited to, the "tetraalkoxysilane" described in Aspect 4 and the nano-sized silica described in Aspect 5. On the other hand, Test Example 4 of JP 2019-123773 A discloses particles equivalent to the nano-sized particles, and the particles contain the anticancer drug sorafenib. However, this anticancer drug alters the properties of the nano-sized particles, and therefore is not a component that can be included in Aspect 1. "Consist" or "consisting of" means that only the components described can be included.

[0038] The "base addition salt" in the "graft polymer or its base addition salt" refers to a salt form formed when the carboxyl groups present in the graft polymer are paired with an alkali or base capable of neutralizing the acidity derived therefrom. Such alkali or base includes alkali metals, ammonia, aliphatic primary amines, for example, C １ ~C １２ alkylamines, basic amino acids such as lysine, ornithine, and arginine.

[0039] The terms "alkyl," by themselves or as part of another substituent, mean, unless structurally inconsistent with the compound or portion thereof in which they are used, a straight-chain, branched-chain, or cyclic saturated hydrocarbon radical, or combinations thereof, having the specified number of carbon atoms (e.g., C １ -C １２ By alkyl is meant alkyl having 1 to 12 carbon atoms. Non-limiting examples of this include groups such as methyl, ethyl, n-propyl, isopropyl, n-butyl, t-butyl, isobutyl, sec-butyl, cyclohexyl, (cyclohexyl)methyl, cyclopropylmethyl, homologs and isomers thereof (e.g., n-pentyl, n-hexyl, n-heptyl, n-octyl, etc.).

[0040] The terms "nano-sized" or "nano" are not limited to the nano-sized range, but refer to, for example, an average diameter of 5 nm to 500 nm, preferably 10 nm to 100 nm, and more preferably 30 nm to 100 nm, and nano-sized micelles or nanoparticles are used interchangeably. The size is the average diameter value that can be determined by dynamic light scattering (DLS) in an aqueous medium (water or physiological saline).

[0041] Hereinafter, nano-sized polymeric micelles or nano-sized particles will mainly be referred to as nanoparticles. The residue of a cyclic nitroxide radical compound in the definition of the repeating unit in (1) defined for Formula I above is known to be useful for preventing or treating diseases and the like that are considered to be caused by the production or presence of excessive reactive oxygen species in a living body, by utilizing the redox function that the nitroxide radical can exhibit in a living body environment when polymerized (see, for example, the various documents cited in the aforementioned WO 2009 / 133647 A1 or Biomaterials '053).

[0042] Without being bound by theory, the graft polymer represented by formula I according to the present invention is different from the block copolymer described in WO 2009 / 133647 A1 not only in the category of polymer but also in that the average particle size of nano-sized micelles or particles can be controlled by, for example, selecting the number of PEG chains, and R ｓｉ Since the residues are covalently bonded to the basic skeleton of the graft polymer, chemical cross-linking is formed via these residues when nanoparticles are formed from the graft polymer, improving stability in aqueous media or body fluids. Furthermore, when the nanoparticles are orally administered, nanomicelles can be provided that reduce the migration of nanoparticles or their constituents into the blood and extend the residence time in the digestive tract. For details of these matters, see Item 3.2. siSMAPo in 3. Results of Biomaterials '053. ＴＮ Pharmacokinetic study of siSMAPo ＴＮ ) can be referred to. The chemical crosslinking can also be carried out, if necessary, through a so-called sol-gel reaction in the presence of tetraalkoxysilane or nano-sized silica, for example, at 10°C to 30°C, which makes it possible to adjust the size of nano-sized core-shell micelles having a silica-containing core and a shell mainly composed of PEG chains, and is useful for providing nanoparticles with improved properties as described above.

[0043] In the copolymer of Formula I, a+b+c+d and n are not limited to, but may be derived from the number of units constituting each repeating unit in a random copolymer, preferably an alternating copolymer, obtained by radical copolymerization of maleic anhydride and styrene, which is the starting material for the copolymer of Formula I. For example, a+b+c+d(m) is an integer of 8 to 1400, or 8 to 200, or 15 to 50; n is an integer of 8 to 1400, or 20 to 300, or 30 to 100; and a+b+c+d:n is 1:1 to 5, or 1:1 to 3. , or in a ratio of 1:1-2, a+b+d:c is in a ratio of 1-60:1, or 1-40:1, or 1-20:1, a+b+c+d is a set of repeating units that can be represented as a whole by m, and the repeating units of m and repeating units with the subscript n are mutually randomly included, and in m, repeating units with the symbols a, b, c, and d are each randomly included, with the proviso that the repeating units with the symbol d are zero (or not present), or, if present, can account for up to 15% of the total number of a+ab+c+d.

[0044] The following description of Preparation Example 1 or a substantially equivalent example thereof in Biomaterials '053, Item 2, 2. siSMAPo ＴＮ Preparation of siSMAPo ＴＮ ), in the graft polymer of Formula I, a, b, c, d, and n can be more specifically defined as follows: n is an integer of 16 to 500, or 25 to 250, or 40 to 160, or 50; a is an integer of 6 to 210, or 8 to 60, or 10 to 40, or about 20; b is an integer of 2 to 42, or 2 to 20, or 3 to 16, or about 4; c is 1 to 20, or 2 to 15, or 2 to 10, or about 2; and d is 0 to 20, or 0 to 16, or 0 to 10, or 0.

[0045] The ratio of each side chain group or graft group, etc., in these repeating units can be adjusted by using various stoichiometric excesses of the starting compound, such as the side chain group or graft group, to be introduced relative to the maleic anhydride (or succinic anhydride) unit in the starting poly(maleic anhydride-styrene) copolymer. For example, when introducing one or both of the residues of TEMPO defined in (1) above and the alkoxysilane group defined in (2) above into the cyclic anhydride residue, selecting an equivalent ratio of the former starting compound to the latter of 20 or more to 1 allows the former residue to be introduced into substantially all of the cyclic anhydride residues. Selecting an equivalent ratio of 10 or less to 1 typically allows the introduction rate to be less than 100%, for example, 10 to 60%. For specific introduction rates, see the production examples in the above publications, which describe specific examples of the introduction of individual groups. Briefly, the product can be produced according to the reaction scheme for Production Example 1 described below.

[0046] L as defined above in (3) for formula I ２ When a protected formyl group defined as A1 or a specific protected formyl group defined as A2 of A in -PEG-A is present at the PEG chain terminus, a targeting ligand or the like can be introduced to the PEG chain terminus via the group. In particular, the latter group can easily bind to amino groups on the surface of mucous membranes or organs, thereby contributing to improving the local retention in vivo of the copolymer of Formula I.

[0047] The graft polymers of Formula I described above can be formed into nanoparticles by dialyzing a solution of the polymer in a water-miscible organic solvent (such as tetrahydrofuran (THF) or methanol) against water through a dialysis membrane, or by ultrafiltration of the aqueous solution after removing the organic solvent. The nanoparticles exhibit an average particle size of about 10 nm to about 600 nm, preferably about 15 nm to about 150 nm, and more preferably about 30 nm to about 100 nm, as measured by dynamic light scattering (DLS) in the form of an aqueous solution or dispersion. The size can be adjusted by, but is not limited to, the number of PEG chains, the content of silica or tetraalkoxysilane, and the like.

[0048] The resulting nano-sized particles or micelles can be isolated by separation means such as centrifugation and can be stored as a dry composition by lyophilization and reconstituted in an aqueous medium as needed.

[0049] <Pharmaceutical Formulations> The nanoparticles prepared as described above can be used as the active ingredient of the present invention to prepare a preventive or therapeutic agent for cancer cachexia or sarcopenia without incorporating a specific drug. Because the nanoparticles are soluble in water and can be reconstituted with water, the preventive or therapeutic agent can take various forms, particularly as an oral formulation. When preparing oral formulations or compositions, conventional pharmaceutical vehicles (e.g., water, glycols, oils, alcohols, flavorings, preservatives, coloring agents, etc.; for oral liquid formulations, e.g., suspensions, elixirs, and solutions) or carriers (e.g., starch, sugars, microcrystalline cellulose, diluents, granules, lubricants, binders, disintegrants, etc.; for oral solid formulations, e.g., powders, hard and soft capsules, and tablets) can be used. For example, when the oral dosage form is a tablet, it may contain a binder (e.g., tragacanth gum, acacia, corn starch, or gelatin), an excipient (e.g., disodium phosphate), a disintegrant (e.g., corn starch, potato starch, alginic acid), a lubricant (e.g., sodium stearate), or a sweetener (e.g., sucrose, lactose, or saccharin). Furthermore, since the nano-sized particles can be dissolved in an aqueous medium, they are also suitable as parenteral administration agents for intravenous injection, embedding in a local area of ​​the body, etc. As described above, in oral administration, Therefore, even if the nano-sized particles are distributed only in the intestinal tract, they can suppress systemic inflammation, etc., and therefore, even when parenterally administered, they can prevent or treat cancer cachexia or sarcopenia. Patients or individuals to whom such administration is to be given include, but are not limited to, cancer patients, particularly those with advanced gastrointestinal cancer or lung cancer, as well as cancer patients who have already developed cancer cachexia as a complication, and individuals or patients who have experienced or are at risk of experiencing a decline in skeletal muscle mass and strength due to aging.

[0050] Such formulations or compositions contain the nanoparticles as an active ingredient in an amount of at least 5% by weight. The percentage of the active ingredient in these formulations or compositions can vary and conveniently be between about 30% and about 90% by weight based on the total weight of the formulation or composition. The nanoparticles may be effective over a wide range of dosages; for example, a dose of about 0.01 mg / day to about 10,000 mg / day may be required for the treatment of an adult. More specifically, a dose of about 0.05 mg / day to about 1,000 mg / day, or about 0.1 mg / day to about 100 mg / day, can be used. The optimal dosage can be determined depending on the desired therapy, the mode of administration, the subject to be treated, the subject's weight, and the experience of the attending physician.

[0051] The present invention will be explained in more detail below with reference to specific examples, but is not limited to these.

[0052] Production Example 1: Production of graft polymer and preparation of nanoparticles from the graft polymer The graft polymer was produced according to the following reaction scheme. The resulting graft polymer (having two grafted PEG chains and R ｓｉ Group and L １ Approximately 4 and 20 groups corresponding to -TEMPO were introduced, respectively. ＴＮ See Figure 1 for a schematic diagram of the formation of

[0053] <Reaction scheme>

[0054] Poly(ethylene glycol) (CH ３O-PEG-OH (5 kDa, 15 g) was dissolved in tetrahydrofuran (THF, 50 mL) at 55°C, and butyllithium (1.6 M, 2.14 mL) was added. This solution was added to a solution of styrene-maleic anhydride copolymer (PSMA: 7.5 kDa, 22.5 g, styrene:maleic anhydride unit ratio = 67:33, obtained from Polysciences, Inc.) in THF (50 mL), and the reaction was carried out at 55°C for 30 minutes. The reaction solution was poured into t-butyl methyl ether (1 L) to precipitate the product. After filtration, the product was washed twice with t-butyl methyl ether to obtain PSMA-g-PEG (33 g).

[0055] PSMA-g-PEG (18.1 g) was added to 180 ml of methanol and stirred. Aminopropyltrimethoxysilane (APTS, 1.31 mL) and 4-amino-2,2,6,6-tetramethylpiperidine 1-oxyl free radical (NH ２ A solution of PSMA-g-PEG (5 g of TEMPO) in 20 mL of methanol was added to the PSMA-g-PEG solution and reacted at 40°C for 1 hour with stirring to give APTS residues and NH ２ A copolymer (siPSMA(T)-g-PEG) with TEMPO residues introduced therein was formed. A water / methanol mixed solution (1 / 1, 150 mL) was added to the reaction solution and the reaction was continued for another 2 hours at 40°C. After the reaction, the methanol was removed using an evaporator, and then the mixture was subjected to ultrafiltration (AKTA flux) to obtain self-assembled particles (siSMAPo ＴＮ The TEMPO concentration in the particle-containing solution was measured by an electron spin resonance (ESR) spectrometer, and the solution was diluted to 5 mM and used in the following tests.

[0056] Test Example 1: A mouse cancer cachexia model was developed by injecting mouse colon cancer cells (C26) at 1x10 ６ The treatment group received siSMAP from the day of cancer cell transplantation. ＴＮ The mice were given the solution via free access to water. Starting from 7 days after cancer cell transplantation, tumor size, body weight, and muscle strength were measured. Hereinafter, the day when measurements began is designated as day 0.

[0057] (1) The changes in body weight are shown in Figure 2. In the figure, ●: tumor-free, ▲: tumor-bearing mice (untreated group), ■: tumor-bearing mice (siSMAPo ＴＮ The graph shows the results for the test group (the group in which the cancer was transplanted) and the group in which the tumor was not treated. It can be understood that the cancer transplant group showed a higher value than the group without tumors because of the presence of tumors.

[0058] (2) The results of the changes in tumor volume are shown in Figure 3. In the figure, ▲: tumor-bearing mice (untreated group), ■: tumor-bearing mice (siSMAPo ＴＮ The figure shows the results of the test group (siSMAPo administration group). ＴＮ No tumor growth effect was observed in the treated group either.

[0059] (3) The results of muscle strength measurements are shown in Figure 4. In the figure, the muscle strength on the day of cancer transplantation is used as the standard, and the fluctuations in each mouse are graphically displayed. ●: tumor-free, ▲: cancer-bearing mice (untreated group), ■: cancer-bearing mice (siSMAPo ＴＮ The figure shows the test results for the siSMAPo administration group. ＴＮ A significant trend of improvement was confirmed in the treatment group compared with the untreated group.

[0060] Test Example 2: On the 21st day after the start of the measurements in Test Example 1, the mice were anesthetized and their blood and quadriceps muscles were collected. Interleukin 6 (IL-6) in the blood is a typical inflammatory cytokine, and its levels have been confirmed to be elevated in patients with cancer cachexia.

[0061] (1) Therefore, siSMAPo ＴＮ The administration groups were examined for changes in blood IL-6 levels. Blood was collected from the mice in heparinized tubes to obtain plasma components. The IL-6 levels in the plasma were measured using a mouse IL-6 ELISA kit (Sigma RAB308). The measurement results are shown in Figure 5. As can be seen from the figure, the amount of IL-6 in the blood increased in the cancer-implanted mice, but the amount of IL-6 in the siSMAPo ＴＮ It can be seen that the increase is significantly suppressed by administration.

[0062] (2) The weight of the collected quadriceps muscle was measured. The weight was reduced in the cancer-implanted mice, and the measurement results are shown in Figure 6. ＴＮ No significant decrease was observed in the administered group, indicating that muscle mass was maintained.

[0063] (3) Atrogin 1 is a typical ubiquitin ligase in muscle cells and plays a central role in muscle protein degradation. Muscles were collected and the expression level of Atrogin 1 was analyzed by Western blotting. The results are shown in Figure 7. In the figure, (a) is a photograph of the Western blotting data, and (b) is the analysis result. As can be seen from the figure, the expression level of Atrogin 1 was increased in the cancer transplant model mice, but the expression level of siSMAPo ＴＮ It can be seen that the increase is suppressed in the administration group.

Claims

1. A preventive or therapeutic agent for cancer cachexia or sarcopenia, comprising as an active ingredient nano-sized particles essentially consisting of a graft polymer represented by the following formula I or a base addition salt thereof. Formula I: In the formula, a+b+c+d is an integer between 8 and 1400, n is an integer between 8 and 1400, a+b+c+d:n is in a ratio of 1:1 to 5, a+b+d:c is in a ratio of 1 to 60:1, a+b+c+d is a set of repeating units that can be represented as a whole by m, the repeating units of m and the repeating units with the subscript n are mutually included randomly or alternately, and in m, the repeating units with the symbols a, b, c, and d are each included randomly, with the proviso that the repeating units with the symbol d are zero (or not present), or, if present, can account for up to 15% of the total number of a+b+c+d. (1) In the repeating units with the subscript a, L １ -TEMPO, L １ is NH or O, and TEMPO is a residue of a cyclic nitroxide radical compound selected from the group consisting of 2,2,6,6-tetramethylpiperidine-1-oxyl-4-yl, 2,2,5,5-tetramethylpyrrolidin-1-oxyl-3-yl, 2,2,5,5-tetramethylpyrrolin-1-oxyl-3-yl, 2,4,4-trimethyl-1,3-oxazolidine-3-oxyl-2-yl, 2,4,4-trimethyl-1,3-thiazolidine-3-oxyl-2-yl, and 2,4,4-trimethyl-imidazoline-3-oxyl-2-yl. (2) In the repeating unit with the subscript b, R ｓｉ is expressed as follows: where R ３ is C １ -C ３ where r is an integer from 1 to 6, and H in NH is C １ -C ６ (3) In the repeating unit marked with the subscript c, L ２ -PEG-A, L ２ is O or NR, where R is a hydrogen atom or C １ -C ６ and PEG is represented by the formula: where p is an integer from 1 to 6, q is an integer from 5 to 500, and A is A1: unsubstituted or substituted C １ -C １２ represents an alkoxy group, and when substituted, the substituent is a formyl group, ａ R ｂ CH- (where R ａ and R ｂ is independent, C １ -C ４ Alkoxy or R １ and R ２ Let's get together - OCH ２ CH ２ O-, -O(CH ２ ) ３ O- or -O(CH ２ ) ４ A2: a group of the following formula: (4) In the repeating unit with the subscript d, R １ and R ２ together represent --O-- to form a cyclic anhydride residue, or each represent OH.

2. A preventive or therapeutic agent for cancer cachexia or sarcopenia as described in claim 1, which is administered orally.

3. A preventive or therapeutic agent for cancer cachexia or sarcopenia according to claim 1, wherein the nano-sized particles are composed of a graft polymer represented by formula I or a base addition salt thereof.

4. The preventive or therapeutic agent for cancer cachexia or sarcopenia according to claim 1, wherein the nano-sized particles are R in the repeating unit marked with the subscript b. ｓｉ and a tetraalkoxysilane (wherein the alkoxy is C) for forming a chemical crosslink with １ -C ４ The aforementioned preventive or therapeutic agent, which comprises a component derived from a compound represented by the formula (I) which is an alkyloxy group.

5. The preventive or therapeutic agent for cancer cachexia or sarcopenia according to claim 1, wherein the nano-sized particles are R in the repeating unit marked with the subscript b. ｓｉ or R ｓｉ The preventive or therapeutic agent as described above, which comprises nano-sized silica that is adsorbed via the 6. The preventive or therapeutic agent for cancer cachexia or sarcopenia according to claim 1, wherein the TEMPO in the repeating unit marked with subscript a is represented by the following formula: wherein R' is a methyl group.

7. Nano-sized particles essentially consisting of a graft polymer represented by the following formula I or a base addition salt thereof for use in the prevention or treatment of cancer cachexia or sarcopenia. Formula I: In the formula, a+b+c+d is an integer between 8 and 1400, n is an integer between 8 and 1400, a+b+c+d:n is in a ratio of 1:1 to 5, a+b+d:c is in a ratio of 1 to 60:1, a+b+c+d is a set of repeating units that can be represented as a whole by m, the repeating units of m and the repeating units with the subscript n are mutually randomly included, and in m, the repeating units with the symbols a, b, c, and d are each randomly included, with the proviso that the repeating units with the symbol d are zero (or not present), or, if present, can account for up to 15% of the total number of a+b+c+d. (1) In the repeating unit with the subscript a, L １ - In TEMPO, L １ is NH or O, TEMPO is a residue of a cyclic nitroxide radical compound selected from the group consisting of 2,2,6,6-tetramethylpiperidine-1-oxyl-4-yl, 2,2,5,5-tetramethylpyrrolidin-1-oxyl-3-yl, 2,2,5,5-tetramethylpyrrolin-1-oxyl-3-yl, 2,4,4-trimethyl-1,3-oxazolidine-3-oxyl-2-yl, 2,4,4-trimethyl-1,3-thiazolidine-3-oxyl-2-yl, and 2,4,4-trimethyl-imidazoline-3-oxyl-2-yl. ｓｉ is expressed as follows: where R ３ is C １ -C ３ where r is an integer from 1 to 6, and H in NH is C １ -C ６ (3) In the repeating unit marked with the subscript c, L ２ -PEG-A, L ２ is O or NR, where R is a hydrogen atom or C １ -C ６ and PEG is represented by the formula: where p is an integer from 1 to 6, q is an integer from 5 to 500, and A is A1: unsubstituted or substituted C １ -C １２ represents an alkoxy group, and when substituted, the substituent is a formyl group, ａ R ｂ CH- (where R ａ and R ｂ is independent, C １ -C ４ Alkoxy or R １ and R ２ Let's get together - OCH ２ CH ２ O-, -O(CH ２ ) ３ O- or -O(CH ２ ) ４ A2: a group of the following formula: (4) In the repeating unit marked with the subscript d, R １ and R ２ together represent --O-- to form a cyclic anhydride residue, or each represent OH.

8. A method for preventing or treating cancer cachexia or sarcopenia, comprising the step of administering an effective amount of nano-sized particles consisting essentially of a graft polymer represented by the following formula I or a base addition salt thereof to a patient suffering from or likely to suffer from cancer cachexia or sarcopenia: Formula I: In the formula, a+b+c+d is an integer between 8 and 1400, n is an integer between 8 and 1400, a+b+c+d:n is in a ratio of 1:1 to 5, a+b+d:c is in a ratio of 1 to 60:1, a+b+c+d is a set of repeating units that can be represented as a whole by m, the repeating units of m and the repeating units with the subscript n are mutually randomly included, and in m, the repeating units with the symbols a, b, c, and d are each randomly included, with the proviso that the repeating units with the symbol d are zero (or not present), or, if present, can account for up to 15% of the total number of a+b+c+d. (1) In the repeating unit with the subscript a, L １ - In TEMPO, L １ is NH or O, and TEMPO is a residue of a cyclic nitroxide radical compound selected from the group consisting of 2,2,6,6-tetramethylpiperidine-1-oxyl-4-yl, 2,2,5,5-tetramethylpyrrolidin-1-oxyl-3-yl, 2,2,5,5-tetramethylpyrrolin-1-oxyl-3-yl, 2,4,4-trimethyl-1,3-oxazolidine-3-oxyl-2-yl, 2,4,4-trimethyl-1,3-thiazolidine-3-oxyl-2-yl, and 2,4,4-trimethyl-imidazoline-3-oxyl-2-yl. ((2) In the repeating unit with the subscript b, R ｓｉ is expressed as follows: where R ３ is C １ -C ３ where r is an integer from 1 to 6, and H in NH is C １ -C ６ (3) In the repeating unit marked with the subscript c, L ２ -PEG-A, L ２ is O or NR, where R is a hydrogen atom or C １ -C ６ and PEG is represented by the formula: where p is an integer from 1 to 6, q is an integer from 5 to 500, and A is: A1: unsubstituted or substituted C １ -C １２ represents an alkoxy group, and when substituted, the substituent is a formyl group, ａ R ｂ CH- (where R ａ and R ｂ is independent, C １ -C ４ Alkoxy or R １ and R ２ Let's get together - OCH ２ CH ２ O-, -O(CH ２ ) ３ O- or -O(CH ２ ) ４ A2: a group of the following formula: (4) In the repeating unit marked with the subscript d, R １ and R ２ together represent --O-- to form a cyclic anhydride residue, or each represent OH.

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