Exercise effect-mimicking agent

Abstract

The present disclosure aims to provide an exercise effect-mimicking agent capable of producing effects similar to those of exercise without actually exercising. An exercise effect-mimicking agent containing at least one compound selected from the group consisting of (A) an α-adrenergic antagonist, (B) a flavonoid, (C) a dopamine D2 receptor antagonist, and (D) an NF-κB signaling pathway inhibitor.

Description

Exercise effect mimicking agent

[0001] The present disclosure relates to an exercise effect mimicking agent and the like.

[0002] In modern society, due to work busyness, stress, etc., people are lacking in exercise, and the risk of lifestyle diseases is increasing. Lifestyle diseases include obesity, hypertension, diabetes, etc., and all of them can be prevented or improved by reviewing daily habits. Exercise significantly promotes overall health and affects many parts of the body. That is, even without administering or ingesting drugs, exercise is known to have non-pharmacological therapeutic effects on various types of diseases other than those called the above-mentioned lifestyle diseases, such as neurodegenerative diseases, musculoskeletal disorders, cardiovascular diseases, etc. Therefore, exercise can be regarded as a drug that combines a plurality of different drugs into one tablet, so-called "polypill". And in recent years, evidence supporting the effectiveness of exercise in skeletal muscle, circulatory system, immune system, brain nerves, bones, cardiovascular system, abdominal organs, etc. is increasing. However, despite the many advantages of exercise, the elderly, patients, etc. often have difficulty participating in exercise. Also, there are people who have difficulty performing exercise such as physical activities, such as those who dislike exercise or those who cannot exercise due to physical problems.

[0003] Therefore, the development of exercise mimicking drugs (exercise pills) that can obtain the same effects as exercising without actually exercising has been carried out. For example, AMP-activated protein kinase (AMPK), activator AICAR, peroxisome proliferator, activator receptor delta (PPARδ) receptor agonist GW501516, REV-ERBα agonist, SR9009, and sirtuin 1 (SIRT1) activator are known to be compounds that activate specific molecular targets in skeletal muscle stimulated by exercise (for example, Non-Patent Document 1, 2, etc.). However, these compounds can only reproduce specific aspects of exercise, and moreover, they tend to over-activate specific molecules, so there are also known compounds with side effects such as carcinogenesis and lactic acidosis (for example, Non-Patent Document 3, etc.). Therefore, the development of compounds with high safety and having an exercise effect mimicking action without exercising has attracted attention.

[0004] W. Fan, et al., ERRγ promotes angiogenesis, mitochondrial biogenesis and oxidative remodeling in PGC1α / β-deficient muscle., Cell Rep., 2018, Volume 22, Issue 10, p2521-2529.E. Woldt, et al., Rev-erb-α modulates skeletal muscle oxidative capacity by regulating mitochondrial biogenesis and autophagy. Nat. Med., 2013, Aug;19(8), p.1039-46.W. Fan, RM Evans, Exercise mimetics: Impact on health and performance. Cell Metab., 2017, Feb 7;25(2), p.242-247.

[0005] This disclosure aims to provide an exercise effect mimic agent that can produce effects similar to those of exercise without actually exercising.

[0006] To solve the above problems, the inventors conducted extensive research and discovered that administering a specific compound from among existing drugs or compounds to mice can produce desirable biological responses that are normally observed during exercise, without the mice having to exercise themselves. This led to the completion of this disclosure.

[0007] In other words, the present disclosure is as follows: Item 1. An exercise effect mimicking agent comprising at least one compound selected from the group consisting of (A) an α-adrenergic antagonist, (B) a flavonoid, (C) a dopamine D2 receptor antagonist, and (D) an NF-κB signaling pathway inhibitor. Item 2. The exercise effect mimicking agent according to Item 1, wherein the exercise effect mimicking agent comprises as an active ingredient a compound that mimics an exercise reference signature, which is a change in the gene expression pattern that occurs in vivo due to exercise. Item 3. The exercise effect mimicking agent according to Item 2, wherein the active ingredient is a compound that shows a connectivity score of 50 or more with respect to the exercise reference signature, which is a change in the gene expression pattern that occurs in vivo due to exercise, based on CMap (Connectivity Map) analysis. Item 4. An exercise effect mimic according to item 2 or 3, wherein the exercise reference signature, which is a change in the gene expression pattern produced in vivo by the exercise, is determined by multi-omics analysis including RNA sequencing analysis. Item 5. An exercise effect mimic according to any one of items 1 to 4, which mimics the effects of aerobic exercise and / or resistance exercise (muscle strengthening exercise). Item 6. An exercise effect mimic according to any one of items 1 to 5, wherein the exercise effect mimic is the (A) α-adrenergic antagonist, and the (A) α-adrenergic antagonist is at least one compound selected from the group consisting of doxazosin, prazosin, terazosin, tamsulosin, silodosin, alfuzosin, naphtopidil, bunazosin, indramin, urapidil, phentolamine, trazoline, phenoxybenzamine, and dibenamine. Item 7. The exercise effect mimicry agent according to any one of claims 1 to 5, wherein the exercise effect mimicry agent is doxazosin and mimics the effects of resistance exercise. Claim 8. The exercise effect mimicry agent according to any one of claims 1 to 5, wherein the exercise effect mimicry agent is the (B) flavonoid, and the (B) flavonoid contains a flavone as an active ingredient. Claim 9. The exercise effect mimicry agent according to claim 8, wherein the flavone is apigenin and mimics the effects of aerobic exercise.Item 10. The exercise effect mimicry agent according to any one of items 1 to 5, wherein the exercise effect mimicry agent is the (C) dopamine D2 receptor antagonist, and the (C) dopamine D2 receptor antagonist is at least one compound selected from the group consisting of sulpiride, amisulpiride, levosulpiride, tiapride, metoclopramide, rimosapride, mosapride, and nemonapride. Item 11. The exercise effect mimic agent is the (D) NF-κB signaling pathway inhibitor, and the (D) NF-κB signaling pathway inhibitor is BMS-345541 (CAS number: 445430-58-0), Bay11-7082 (CAS number: 19542-67-7), IMD-0354 (CAS number: 978-62-1), IMD-1041 (CAS number: 10736) An exercise effect mimic agent according to any one of claims 1 to 5, which is at least one selected from the group consisting of 66-73-5), IMD-2560, TPCA1 (CAS number: 507475-17-4), BOT-64 (CAS number: 113760-29-5), SC-514 (CAS number: 354812-17-2), IKK-16 (CAS number: 1186195-62-9), ertiprotafib (CAS number: 251303-04-5), and Bay65-1942 (HCl salt) (CAS number: 600734-06-3). Claim 12. A skeletal muscle function enhancer comprising an exercise effect mimic agent according to any one of claims 1 to 5. Claim 13. An energy metabolism enhancer comprising an exercise effect mimicking agent described in any one of items 1 to 5. Item 14. A chondrocyte proliferation or differentiation inducer comprising an exercise effect mimicking agent described in any one of items 1 to 5. Item 15. A gene expression enhancer comprising an exercise effect mimicking agent described in any one of items 1 to 5, wherein the gene whose expression is to be promoted is a gene that improves mitochondrial function in muscle tissue. Item 16. The gene expression enhancer according to item 15, wherein the gene that improves mitochondrial function in muscle tissue is PGC-1α. Item 17. An oral composition or food / beverage composition comprising an exercise effect mimicking agent described in any one of items 1 to 5. Item 18. A pharmaceutical composition comprising an exercise effect mimicking agent described in any one of items 1 to 5.Item 19. A supplement containing an exercise effect mimic agent as described in any one of items 1 to 5.

[0008] Item 1A. An exercise effect mimicry composition comprising at least one compound selected from the group consisting of (A) an α-adrenergic antagonist, (B) a flavonoid, (C) a dopamine D2 receptor antagonist, and (D) an NF-κB signaling pathway inhibitor. Item 2A. The composition according to Item 1A, wherein the composition contains as an active ingredient a compound having a connectivity score greater than 0 in gene expression signature matching analysis (including CMap (Connectivity Map) analysis) with respect to an exercise reference signature (a reference signature based on multi-omics analysis including RNA sequencing analysis of skeletal muscle after exercise intervention). Item 3A. The composition according to Item 2A, wherein the connectivity score is 50 or greater. Item 4A. The composition according to Item 2A, wherein the exercise reference signature is determined by multi-omics analysis including RNA sequencing analysis. Item 5A. The composition according to claim 2A, wherein the composition mimics the effects of aerobic exercise and / or resistance exercise. Claim 6A. The composition according to claim 1A, wherein (A) comprises at least one selected from doxazosin, prazosin, terazosin, tamsulosin, alfuzosin, silodosin, indramin, naphtopidil, urapidil and bunazosin. Claim 7A. The composition according to claim 1A, wherein (B) comprises a flavone, the flavone being apigenin. Claim 8A. The composition according to claim 1A, wherein (C) is at least one selected from sulpiride, amisulpiride, levosulpiride, tiapride, metoclopramide, rimosapride, mosapride and nemonapride. Claim 9A. The composition according to claim 1A, wherein (D) is at least one selected from BMS-345541, BAY11-7082, IMD-0354, IMD-1041, IMD-2560, TPCA-1, BOT-64, SC-514, IKK-16, ertiprotafib and BAY65-1942 (HCl salt). Claim 10A. The composition according to claim 1A, wherein the composition is an oral composition, a food or beverage composition or a pharmaceutical composition. Claim 11A. A method for mimicking the effects of exercise in a subject, comprising administering a therapeutically effective amount of a compound in the composition according to claim 1A to a subject in need thereof.Item 12A. The method according to item 11A, wherein the composition exhibits a positive connectivity score to an exercise reference signature evaluated by an exercise multi-omics signature evaluation method. Item 13A. The method according to item 12A, wherein the exercise reference signature includes a group of hub genes identified after exercise, and the positive connectivity score to the hub gene group is 50 or higher. Item 14A. The method according to item 12A, wherein the compound is a (B) flavonoid containing apigenin, and the exercise reference signature mimics an aerobic exercise-like signature. Item 15A. The method according to item 12A, wherein the compound is an (A) α-adrenergic antagonist containing doxazosin, and the exercise reference signature mimics a resistance exercise-like signature. Item 16A. A method for improving skeletal muscle function, promoting energy metabolism, or preventing muscle atrophy in the subject, comprising administering a therapeutically effective amount of the compound in the composition described in item 1A to the subject. Item 17A. A method for identifying a compound having the ability to mimic the effects of exercise, comprising: obtaining a gene expression profile induced by a candidate compound; comparing the gene expression profile with an exercise reference signature obtained from skeletal muscle after exercise intervention; and selecting the candidate compound as a compound having exercise effect mimicry when it shows a positive connectivity score to the exercise reference signature. Item 18A. The method according to item 17A, wherein the exercise reference signature includes a gene expression pattern obtained by RNA-seq analysis of skeletal muscle after aerobic exercise intervention and / or resistance exercise intervention. Item 19A. The method according to item 17A, wherein the exercise reference signature includes a group of hub genes identified by network analysis of the RNA-seq analysis. Item 20A. The method according to claim 17A, wherein the selection step includes selecting the candidate compound when the connectivity score is 50 or higher.

[0009] Item 1B. A motor effect mimicking agent comprising at least one compound selected from the group consisting of (A) an α-adrenergic antagonist, (B) a flavonoid, (C) a dopamine D2 receptor antagonist, and (D) an NF-κB signaling pathway inhibitor. Item 2B. The agent according to Item 1B, wherein the agent comprises a compound having a connectivity score of at least 50 to a motor reference signature based on CMap (Connectivity Map) analysis. Item 3B. The agent according to Item 1B or Item 2B, wherein the motor reference signature is determined by multi-omics analysis including RNA sequencing analysis. Item 4B. The agent according to any one of claims 1B to 3B, wherein (A) is at least one selected from doxazosin, prazosin, terazosin, tamsulosin, alfuzosin, silodosin, indramin, naphtopidil, urapidil, and bunazosin. Claim 5B. The agent according to any one of claims 1B to 4B, wherein (B) comprises a flavone, the flavone being apigenin. Claim 6B. The agent according to any one of claims 1B to 5B, wherein (C) is at least one selected from sulpiride, amisulpiride, levosulpiride, tiapride, metoclopramide, rimosapride, mosapride, and nemonapride. Claim 7B. The agent according to any one of claims 1B to 6B, wherein (D) is at least one selected from BMS-345541, BAY11-7082, IMD-0354, IMD-1041, IMD-2560, TPCA-1, BOT-64, SC-514, IKK-16, ertiprotafib and BAY65-1942 (HCl salt). Claim 8B. An oral composition, food or beverage composition or pharmaceutical composition comprising the agent according to any one of claims 1B to 7B. Claim 9B. An oral composition, food or beverage composition or pharmaceutical composition comprising the agent according to any one of claims 1B to 7B as an active ingredient, for use in improving skeletal muscle function. Claim 10B. An oral composition, food or beverage composition, or pharmaceutical composition containing an active ingredient described in any of items 1B to 7B, for use in the prevention or treatment of disuse muscle atrophy.Item 11B. An oral composition, food or beverage composition, or pharmaceutical composition containing an agent described in any of Items 1B to 7B as an active ingredient, for use in reducing the severity of osteoarthritis of the knee. Item 12B. A food composition containing apigenin described in Item 5B as an active ingredient, for use in improving exercise endurance.

[0010] Item 1C. An exercise effect mimicking agent comprising at least one compound selected from the group consisting of (A) an α-adrenergic antagonist, (B) a flavonoid, (C) a dopamine D2 receptor antagonist, and (D) an NF-κB signaling pathway inhibitor. Item 2C. The exercise effect mimicking agent according to Item 1C, wherein the (A) α-adrenergic antagonist is at least one compound selected from the group consisting of doxazosin, prazosin, terazosin, tamsulosin, silodosin, alfuzosin, naphtopidil, bunazosin, indramin, urapidil, phentolamine, trazoline, phenoxybenzamine, and dibenamine. Item 3C. The exercise effect mimicking agent according to Item 1C, wherein the (B) flavonoid is at least one compound selected from the group consisting of flavones, isoflavones, and flavanols. Item 4C. The exercise effect mimicry agent according to item 1C, wherein the (B) flavonoid is at least one compound selected from the group consisting of apigenin, chrysin, and luteolin. Item 5C. The exercise effect mimicry agent according to item 1C, wherein the (C) dopamine D2 receptor antagonist is at least one compound selected from the group consisting of sulpiride, amisulpiride, levosulpiride, tiapride, metoclopramide, rimosapride, mosapride, and nemonapride. Item 6C. The exercise effect mimicry agent according to item 1C, wherein the (D) NF-κB signaling pathway inhibitor is an IKKβ inhibitor. Item 7C. The aforementioned IKKβ inhibitors are BMS-345541 (CAS number: 445430-58-0), Bay11-7082 (CAS number: 19542-67-7), IMD-0354 (CAS number: 978-62-1), IMD-1041 (CAS number: 1073666-73-5), IMD-2560, TPCA1 (CA An exercise effect mimic agent according to item 6C, which is at least one selected from the group consisting of S number: 507475-17-4, BOT-64 (CAS number: 113760-29-5), SC-514 (CAS number: 354812-17-2), IKK-16 (CAS number: 1186195-62-9), ertiprotafib (CAS number: 251303-04-5), and Bay65-1942 (HCl salt) (CAS number: 600734-06-3).Item 8C. A skeletal muscle function enhancer comprising the exercise effect mimicry agent described in Item 1C. Item 9C. An energy metabolism enhancer comprising the exercise effect mimicry agent described in Item 1C. Item 10C. A chondrocyte proliferation or differentiation inducer comprising the exercise effect mimicry agent described in Item 1C. Item 11C. A gene expression enhancer comprising the exercise effect mimicry agent described in Item 1C, wherein the gene whose expression is promoted is a gene that improves mitochondrial function in muscle tissue. Item 12C. The gene expression enhancer according to Item 11C, wherein the gene that improves mitochondrial function in muscle tissue is PGC-1α. Item 13C. A pharmaceutical product comprising the exercise effect mimicry agent described in Item 1C. Item 14C. An oral composition comprising the exercise effect mimicry agent described in Item 1C. Item 15C. A food and beverage composition comprising the exercise effect mimicry agent described in Item 1C. Item 16C. Use for the manufacture of exercise effect mimics using at least one compound or salt thereof selected from the group consisting of (A) an α-adrenergic antagonist, (B) a flavonoid, (C) a dopamine D2 receptor antagonist, and (D) an NF-κB signaling pathway inhibitor.

[0011] According to this disclosure, it is possible to provide an exercise effect mimic agent that can produce effects similar to those of exercise even without actually exercising.

[0012] Figure 1 shows the results of the Connectivity Map (CMap). Figure 1(1) shows the connectivity scores of the top compounds that cause changes in gene expression in RNA sequencing (hereinafter also referred to as "RNA-seq") analysis after aerobic exercise. Figure 1(2) shows the connectivity scores of the top compounds that cause changes in gene expression in RNA-seq analysis after resistance exercise. Figure 1(3) shows the connectivity scores of the top 10 compounds that cause changes in the hub gene in both RNA-seq analysis and RRBS (Reduced Representation Bisulfite Sequencing) analysis after aerobic exercise. Figure 1(4) shows the connectivity scores of the top 10 compounds that cause changes in the hub gene in both RNA-seq analysis and RRBS analysis after resistance exercise. Figure 1(5) shows the connectivity scores of the top 10 compounds via well-changing genes in RNA-seq analysis after aerobic exercise. Figure 1(6) shows the connectivity scores of the top 10 compounds via well-changing genes in RNA-seq analysis after resistance exercise. Figure 2 is a schematic diagram illustrating the experimental timeline of compound administration in healthy young mice. Figure 3 shows grip strength after administration of low or high concentrations of doxazosin for two weeks. Figure 4 shows grip strength after administration of low or high concentrations of apigenin for two weeks. Figure 5 shows the distance run in an endurance test after administration of low concentrations of sulpiride for two weeks (P = 0.058). Figure 6 shows the tibialis anterior (TA) muscle weight after administration of low or high concentrations of BMS-345541 for two weeks. Figure 7 is a schematic diagram illustrating the experimental schedule using apigenin or doxazosin in combination with moderate exercise. Figure 8 shows the distance run in the endurance test for each group during the 4-week intervention period. Figure 9(1) is a representative image of the Western blotting analysis of PGC-1α and GAPDH in the calf muscle of mice during the apigenin administration experiment, and Figure 9(2) shows the relative quantification of protein expression using the results of Western blotting analysis during the apigenin administration experiment (P = 0.066).Figure 10(1) is a photograph of a representative SDH staining image after apigenin administration (scale bar, 200 μm), and Figure 10(2) is a diagram showing the SDH activity level during the apigenin administration experiment (P = 0.088). Figure 11 is a diagram showing grip strength during the doxazosin administration experiment. Figure 12(1) is a photograph of a representative SDH staining image during the doxazosin administration experiment (scale bar, 200 μm), and Figure 12(2) is a diagram showing the SDH activity level during the doxazosin administration experiment. Figure 13(1) is a photograph of a representative immunohistochemical image of type I and type II fibers of mouse gastrocnemius muscle (scale bar, 100 μm), and Figure 13(2) is a diagram showing the quantification of type II muscle fiber area (S) during the doxazosin administration experiment (P = 0.078). Figure 14 is a schematic diagram showing the experimental timeline of administration of apigenin or doxazosin to HLU mice. Figure 15 shows the soleus muscle weight in the HLU experiment. Figure 16(1) is a photograph of immunohistochemical staining of the gastrocnemius muscle in the HLU experiment, and Figure 16(2) shows the cross-sectional area of ​​type I fibers (P = 0.069). Figure 17(1) is a μCT image (scale bar, 100 μm) in the HLU experiment, Figure 17(2) shows the bone volume / tissue volume ratio (BV / TV) in the HLU experiment, Figure 17(3) shows the number of cancellous bones (Tb.N), and Figure 17(4) shows the cancellous bone spacing (Tb.Sp). Figure 18 is a schematic diagram showing the experimental timeline of administration of apigenin or doxazosin to traumatic osteoarthritis of the knee. Figure 19 is a photograph (scale bar, 1 mm) of safranin-O / fast green (SOFG) stained images in a traumatic osteoarthritis of the knee experiment. Figure 20(1) shows the OARSI score in the traumatic osteoarthritis of the knee experiment, and Figure 20(2) shows the SOFG staining intensity (%).

[0013] Exercise Effect Mimicry Agent The exercise effect mimicry agent of this disclosure contains at least one compound selected from the group consisting of (A) α-adrenergic antagonists, (B) flavonoids, (C) dopamine D2 receptor antagonists, and (D) NF-κB signaling pathway inhibitors. The active ingredient contained in the exercise effect mimicry agent of this disclosure may hereinafter be referred to as "compound (1)". The exercise effect mimicry agent of this disclosure may hereinafter be referred to as "the agent of this disclosure". The exercise effect mimicry agent of this disclosure can use one or more compounds from the compound group (A) to (D) belonging to these categories as components. The exercise effect mimicry agent of this disclosure is a known agent and can be said to be a compound whose safety has been confirmed. In addition, for example, (B) flavonoids are compounds with high safety. In particular, flavonoids contained in food are known to have relatively low toxicity.

[0014] (A) Examples of α-adrenergic antagonists include α1-selective antagonists or non-selective α-antagonists, and specific examples include, but are not limited to, doxazosin, prazosin, terazosin, tamsulosin, silodosin, alfuzosin, naphtopidil, bunazosin, indramin, urapidil, phentolamine, trazoline, phenoxybenzamine, dibenamine, or pharmaceutically acceptable salts thereof.

[0015] In this specification, there are no particular limitations on pharmaceutically acceptable salts, and examples include salts with alkali metals (sodium, potassium, etc.); salts with alkaline earth metals (magnesium, calcium, etc.); salts with organic bases (pyridine, triethylamine, etc.); salts with amines; salts with organic acids (acetic acid, formic acid, propionic acid, fumaric acid, maleic acid, succinic acid, tartaric acid, citric acid, malic acid, oxalic acid, benzoic acid, methanesulfonic acid, etc.); and salts with inorganic acids (hydrochloric acid, phosphoric acid, hydrobromic acid, sulfuric acid, nitric acid, etc.).

[0016] The compound (1) contained in the agent of this disclosure may be in the form of a solvate or in the form of a solvate of a salt of compound (1). The solvate is not particularly limited and examples include hydrates and ethanol solvates.

[0017] In this disclosure, one or more compounds from the group of compounds belonging to these can be used as component (A).

[0018] Alpha-adrenergic antagonists are substances that act on alpha-adrenergic receptors. These receptors are part of the sympathetic nervous system and are known to bind to norepinephrine, adrenaline, etc., to trigger various physiological responses. Alpha-adrenergic receptors are known to play an important role in blood pressure regulation, vasoconstriction, and vasodilation. Alpha-adrenergic antagonists are also called alpha-adrenergic receptor blockers, and alpha-1 adrenergic receptor blockers and alpha-2 adrenergic receptor blockers are known.

[0019] (B) The flavonoid is not particularly limited and includes, for example, flavones, isoflavones, flavanols, flavonols, flavanones, flavonolignans, and pharmaceutically acceptable salts thereof. In this disclosure, one or more compounds belonging to these categories can be used as component (B).

[0020] Flavones are known compounds that have 2,3-didehydroflavan-4-one as their basic structure. Specific examples of flavones include apigenin, chrysin, baicalein, 2,3-didehydroflavan-4-one, luteolinidin, luteolin, apigeninidin, and pharmaceutically acceptable salts thereof. From the viewpoint of further enhancing the exercise effect mimicry effect, apigenin, chrysin, baicalein, etc., are preferred.

[0021] Isoflavones are known compounds that have 3-phenylchromone as their basic structure. Specific examples of isoflavones include daidzein, genistein, and pharmaceutically acceptable salts thereof, with daidzein being preferred.

[0022] Flavanols (flavan-3-ols) are a group of compounds known to have 2-phenyl-3,4-dihydro-2H-chromen-3-ol as their basic structure. Specific examples of flavanols include catechin, epicatechin, epigallocatechin, epicatechin gallate, epigallocatechin gallate, theaflavin, etc., with catechin being preferred.

[0023] Flavonols are a group of compounds known to have 3-hydroxyflavone as their basic structure. Specific examples of flavonols include 3-hydroxyflavone, quercetin, kaempferol, myricetin, and pharmaceutically acceptable salts thereof.

[0024] Flavanones are known compounds (including glycosides) that have 2-phenylchromanone as their basic structure. Specific examples of flavanones include naringenin, hesperidin, naringin, ponsirin, sakuranin, and pharmaceutically acceptable salts thereof.

[0025] Flavonolignans are a group of compounds known for having a basic structure consisting of a flavonoid moiety and a lignan moiety. Specific examples of flavonolignans include silybinin, isosilibinin, silycristin, silydiane, and mixtures thereof such as silymarin, as well as pharmaceutically acceptable salts thereof.

[0026] (C) The dopamine D2 receptor antagonist is not particularly limited, and for example, known dopamine D2 receptor antagonists can be used. Specifically known dopamine D2 receptor antagonists include sulpiride, risperidone, paliperidone, haloperidol, olanzapine, quetiapine, ziprasidone, aripiprazole, iloperidone, fluphenazine, chlorpromazine, morindone, perphenazine, thioridazine, mesolidazine, lacloprid, amisulpiride, domperidone, and pharmaceutically acceptable salts thereof. Among these, the dopamine D2 receptor antagonist is preferably sulpiride, risperidone, paliperidone, aripiprazole, or a salt thereof. In this disclosure, one or more compounds belonging to this group can be used as component (C).

[0027] The dopamine D2 receptor is a type of receptor that binds to the neurotransmitter dopamine. Dopamine D2 receptors are mainly found in the central nervous system and are known to be involved in motor control, motivation, learning, and hormone secretion regulation.

[0028] A "dopamine D2 receptor antagonist" refers to a compound that acts on the dopamine D2 receptor and, as a result, inhibits its function in transmitting signals within cells. More specifically, dopamine D2 receptor antagonists include, but are not limited to, competitive antagonists that bind to the dopamine D2 receptor at the same binding site (active site) as agonists such as dopamine (for example, reversibly) but do not activate the dopamine D2 receptor; non-competitive antagonists that bind to the dopamine D2 receptor at a different binding site than the agonist and allosterically suppress the activation of the dopamine D2 receptor or inhibit the binding of the agonist to the dopamine D2 receptor; non-competitive antagonists that bind to the dopamine D2 receptor to which the agonist has bound and inhibit its activation; partial antagonists; inverse antagonists; etc.

[0029] (D) There are no particular limitations on the NF-κB signaling pathway inhibitors, and examples include IKKβ inhibitors. In this disclosure, one or more compounds belonging to this group can be used as component (D). NF-κB (nuclear factor-kappa B) is a protein complex that acts as a transcription factor and is a hetero or homodimer of a protein belonging to the Rel family. The NF-κB signaling pathway is known to be one of the transcription factors that play an important role in immune responses, inflammatory responses, etc., and this pathway is known to be involved in various biological processes such as cell survival, proliferation, differentiation, inflammation, and stress response. IKKβ inhibitors are drugs that suppress the activity of IκB kinase β (IKKβ). IKKβ is an important component of the NF-κB signaling pathway and is involved in inflammatory responses, immune responses, etc.

[0030] The IKKβ inhibitors mentioned above are not particularly limited and include, for example, BMS345541 (CAS number: 445430-58-0), Bay11-7082 (CAS number: 19542-67-7), IMD-0354 (CAS number: 978-62-1), IMD-1041 (CAS number: 1073666-73-5), IMD-2560, Examples include TPCA1 (CAS number: 507475-17-4), BOT-64 (CAS number: 113760-29-5), SC-514 (CAS number: 354812-17-2), IKK-16 (CAS number: 1186195-62-9), Eltiprotafib (CAS number: 251303-04-5), Bay65-1942 (HCl salt) (CAS number: 600734-06-3), and pharmaceutically acceptable salts thereof.

[0031] The exercise effect mimicry agent of this disclosure contains as an active ingredient a compound that mimics an exercise reference signature, which is a change in the gene expression pattern that occurs in a living organism due to exercise. Specifically, the exercise effect mimicry agent of this disclosure contains as an active ingredient a compound that, based on CMap (Connectivity Map) analysis, exhibits a connectivity score with the exercise reference signature, which is a change in the gene expression pattern that occurs in a living organism due to exercise, that is usually greater than 0 (indicates a positive value), preferably 50 or more, more preferably 70 or more, and particularly preferably 80 or more.

[0032] The movement reference signature, which is the change in gene expression patterns that occurs in vivo due to the aforementioned movement, can be determined by multi-omics analysis, including RNA sequencing analysis. Multi-omics analysis is a method that integrates and analyzes multiple omics information within a living organism, such as genomes, transcripts, proteins, and metabolites. This makes it possible to understand life phenomena or elucidate disease mechanisms in a more comprehensive way than can be achieved with a single omics analysis.

[0033] The exercise multi-omics signature evaluation method is a method for evaluating exercise-specific biological response patterns (signatures) by comprehensively analyzing multiple omics data (genome, transcriptome, proteome, metabolome, etc.) to determine the changes in the body caused by exercise. Gene expression signature matching methods may be utilized in this evaluation method. Gene expression signature matching methods are techniques that identify a series of gene expression patterns (signatures) associated with the state of a specific cell, disease, etc., and search for cells or drugs that have patterns similar to those signatures. Examples of gene expression signature matching methods include, but are not limited to, CMap (Connectivity Map) analysis. Gene expression signature matching methods equivalent to CMap analysis can also be used. The exercise effect mimic agent of this disclosure is confirmed to have a connectivity score greater than 0 (showing a positive value) to the exercise reference signature by the exercise multi-omics signature evaluation method. A preferred connectivity score is 50 or higher, more preferably 70 or higher, and particularly preferably 80 or higher. The upper limit of the connectivity score is 100.

[0034] An exercise reference signature refers to the changes in gene expression patterns that occur in a living organism due to exercise. Specifically, it is fingerprint data that quantifies and directs molecular changes that occur in a living organism (mainly skeletal muscle) due to exercise, using them as a template. It is called a "reference signature" or "reference signature" because it serves as a standard (reference) for matching with compound profiles in CMap, etc. In this specification, "exercise reference signature" refers to a set of genes in which differential expression or differential methylation was observed based on RNA-seq (including RRBS as necessary) data obtained from skeletal muscle after exercise intervention, and is a ranking list (or vector) to which the direction of increase or decrease for each gene and an effect size index are assigned.

[0035] Examples of the exercise reference signatures include signatures obtained from RNA-seq analysis in skeletal muscle after aerobic and / or resistance exercise, signatures containing hub gene groups identified by RNA-seq network analysis, and signatures containing genes that commonly change after exercise as determined by RNA-seq and RRBS. RNA-seq analysis is an analytical method that measures gene expression levels, while RRBS (Reduced Representation Bisulfite Sequencing) is an analytical method that measures epigenetic (DNA methylation) changes. By combining these two methods, important genes can be identified.

[0036] As used herein, a "hub gene" refers to a gene that, in a co-expression or PPI (protein-protein interaction) network constructed based on RNA-seq data after exercise intervention, is in the top X% (e.g., top 5%) of at least one of its degree centrality, betweenness centrality, near-centrality, or eigenvector centrality.

[0037] The Connectivity Score (MCS) in CMap quantifies the similarity between the gene expression changes induced in cells by a specific compound and known gene expression profiles. The score is generally expressed in the range of -100 to 100. A positive value greater than 0 indicates similarity to a reference profile. The exercise effect mimics of this disclosure preferably have a positive Connectivity Score of 50 or higher relative to the exercise reference signature.

[0038] "Moderate exercise" refers to exercise at an intensity that increases the anabolic response of skeletal muscles, for example, aerobic exercise at approximately 70-95% VO2. ２ This includes, but is not limited to, intensities equivalent to the maximum, or in resistance exercises, intensities equivalent to a load of approximately 120% of body weight.

[0039] The exercise effect-mimicking agent of the present disclosure can be used as a skeletal muscle function improver, an energy metabolism promoter, a gene expression promoter, and the like. Further, the exercise effect-mimicking agent of the present disclosure can be used as a skeletal muscle weight increasing agent, a muscle atrophy inhibitor, a preventive or ameliorating agent for muscle atrophy, a muscle strength increasing agent, a preventive or ameliorating agent for bone atrophy, a preventive or severity reducing agent for osteoarthritis deformans, a preventive or ameliorating agent for disuse syndrome, a fat accumulation inhibitor, a heat production promoter, a muscle differentiation promoter, an exercise endurance improver, a skeletal muscle fiber composition changing agent, an energy metabolism enhancer, a basal metabolic rate increasing agent, an organ function improver, and the like.

[0040] The skeletal muscle function improver containing the exercise effect-mimicking agent of the present disclosure contains at least one compound selected from the group consisting of the above (A) α-adrenergic antagonist, (B) flavonoid, (C) dopamine D2 receptor antagonist, and (D) NF-κB signal transduction pathway inhibitor.

[0041] The energy metabolism promoter containing the exercise effect-mimicking agent of the present disclosure contains at least one compound selected from the group consisting of the above (A) α-adrenergic antagonist, (B) flavonoid, (C) dopamine D2 receptor antagonist, and (D) NF-κB signal transduction pathway inhibitor.

[0042] The gene expression promoter containing the exercise effect-mimicking agent of the present disclosure contains at least one compound selected from the group consisting of the above (A) α-adrenergic antagonist, (B) flavonoid, (C) dopamine D2 receptor antagonist, and (D) NF-κB signal transduction pathway inhibitor. The gene in the gene expression promoter is not particularly limited, and examples thereof include PGC-1α and the like.

[0043] The exercise effect-mimicking agent, skeletal muscle function improver, chondrocyte proliferation or differentiation inducer, energy metabolism promoter, or gene expression promoter of the present disclosure can be used for applications such as pharmaceuticals, oral compositions, food and beverage compositions, and the like.

[0044] The "exercise effect mimicking agent" refers to an agent that can achieve the same effects as those obtained from exercising without actually exercising. Here, the exercise effect mimicking agent can be alternatively referred to as an exercise mimicking drug, an exercise effect mimicking drug, an exercise mimicking effect enhancer, an exercise pill, etc.

[0045] More specifically, an agent that can achieve the same effects as those obtained from exercising without actually exercising means an agent that can provide, without performing exercise, the biological responses that are originally observed when exercising, such as improvement of skeletal muscle function, promotion of energy metabolism, promotion of gene expression, increase in skeletal muscle weight, suppression of muscle atrophy, prevention or improvement of muscle atrophy, increase in muscle strength, prevention or improvement of bone atrophy, prevention or reduction of the severity of osteoarthritis, prevention or improvement of disuse syndrome, suppression of fat accumulation, promotion of heat production, promotion of muscle differentiation, improvement of exercise endurance, change in skeletal muscle fiber composition, enhancement of energy metabolism, increase in basal metabolic rate, improvement of the function of the locomotor organs, etc., or can provide the same effects as normal exercise with less exercise.

[0046] Exercise significantly enhances overall health and affects many parts of the body. For example, the parts on which the exercise effect mimicking agent of the present disclosure acts are not particularly limited, and examples include skeletal muscle, circulatory system, immune system, cranial nerves, bone, cardiovascular system, abdominal organs, etc. Therefore, the exercise effect mimicking agent of the present disclosure can improve various symptoms or diseases such as metabolic syndrome and neurodegenerative diseases.

[0047] Exercise can be classified according to various criteria. Generally, examples of the types of exercise include aerobic exercise, anaerobic exercise, etc.

[0048] Aerobic exercise is a type of exercise that improves cardiopulmonary function and includes activities such as running, cycling, and swimming. Aerobic exercise is known to improve endurance and promote heart and lung health. Generally, aerobic exercise is known to enhance mitochondrial function, oxidative phosphorylation capacity, and capillary density. Aerobic exercise significantly enhances estrogen signaling pathways related to mitochondrial biosynthesis and insulin sensitivity, and upregulates major mitochondrial genes, including PGC-1α and NRF1. Here, aerobic exercise is also referred to as running, walking, etc.

[0049] Anaerobic exercise is a type of exercise that strengthens muscles, involving short bursts of high-intensity exercise, or exercise that generates energy without using oxygen. Examples include sprints and high-intensity interval training (HIIT).

[0050] The above-mentioned anaerobic exercises also include resistance exercises. Resistance exercises (sometimes called resistance training) are exercises that strengthen muscles by applying load, and include weightlifting, push-ups, squats, etc. This is often considered a part of anaerobic exercise. Generally, resistance exercises are known to promote protein synthesis, muscle hypertrophy, and muscle strength. Resistance exercises are also called muscle strengthening exercises.

[0051] Exercise intensity is extremely important; below the optimal level, no benefits are obtained, and too much intensity can have adverse effects. Therefore, a "moderate" level of activity is necessary to obtain the maximum benefits.

[0052] In this specification, “moderate” exercise is defined as exercise intensity that optimizes the response of skeletal muscles. Moderate refers to the exercise intensity at which muscle synthesis is maximized in the balance between muscle synthesis and breakdown.

[0053] Skeletal muscle facilitates crosstalk with other organs such as the brain, intestines, bones, and fat through the secretion of myokines.

[0054] PGC-1α is a molecule identified as a transcriptional coactivator that binds to the transcription factor PPARγ, and it regulates the expression of many genes involved in energy production, heat consumption, and other processes. When PGC-1α is introduced into white adipocytes, brown adipocyte-like changes occur, such as enhanced mitochondrial biosynthesis and increased UCP1 expression. PGC-1α activates the transcription factors NRF (nuclear respiratory factor) 1 / NRF2, and NRF activates mitochondrial biosynthesis by promoting the transcription of TFAM (mitochondria transcription factor A). Furthermore, PGC-1α, a central regulator of mitochondrial biosynthesis, controls various reactions in skeletal muscle after aerobic exercise, including angiogenesis, antioxidant activity, gluconeogenesis, lipid metabolism, autophagy, and anti-inflammatory processes. Furthermore, PGC-1α is thought to play an important role in the transformation of skeletal muscle through exercise training, as it activates mitochondrial biosynthesis in skeletal muscle and induces slow-twitch muscle characteristics such as enhanced fatty acid oxidation or an increase in type I MHC (Myosin heavy chain) (Wataru Ogawa et al., Latest Medicine, 70, 2015, pp. 580-586).

[0055] The exercise effect mimicking agent of this disclosure may be a fat accumulation inhibitor, a thermoproduction promoter, a muscle differentiation promoter, a skeletal muscle weight increase agent, an exercise endurance enhancer, a skeletal muscle fiber composition alteration agent, an energy metabolism enhancer, a basal metabolic rate increase agent, a musculoskeletal function improver, or a muscle atrophy inhibitor.

[0056] The effects of the exercise effect mimic agents of this disclosure can be confirmed by examining whether a corresponding biological response is obtained in vivo or in vitro, or by examining the gene expression level.

[0057] The exercise effect mimicking agents of this disclosure have, for example, exercise effect mimicking effects, skeletal muscle function improvement effects, energy metabolism promotion effects, gene expression promotion effects, skeletal muscle weight increase effects, muscle atrophy suppression effects, muscle atrophy prevention or improvement effects, muscle strength increase effects, bone atrophy prevention or improvement effects, knee osteoarthritis prevention or severity reduction effects, disuse syndrome prevention or improvement effects, fat accumulation suppression effects, heat production promotion effects, muscle differentiation promotion effects, exercise endurance improvement effects, skeletal muscle fiber composition change effects, energy metabolism enhancement effects, basal metabolic rate increase effects, and musculoskeletal function improvement effects. Here, "effect" can be rephrased as "effect". These effects can be confirmed, for example, by measuring muscle strength (grip strength), running distance, tibialis anterior (TA) muscle weight, PGC-1α, succinate dehydrogenase (SDH) activity, type II fiber cross-sectional area, soleus muscle weight, type I fiber cross-sectional area, bone volume / tissue volume ratio (BV / TV), cancellous bone number (Tb.N), cancellous bone spacing (Tb.Sp), osteoarthritis severity index (OARSI) score, safranin-O / fast green (SOFG) staining intensity, etc., as shown in the examples below. The agents disclosed herein can be described as muscle strength enhancers, distance-increasing agents, endurance enhancers, tibialis anterior (TA) muscle weight increase agents, PGC-1α expression increase agents, succinate dehydrogenase (SDH) activity enhancers, type II fiber cross-sectional area increase agents, soleus muscle weight loss inhibitors, soleus muscle weight increase agents, type I fiber cross-sectional area increase agents, bone volume / tissue volume ratio (BV / TV) increase agents, cancellous bone number (Tb.N) increase agents, cancellous bone spacing (Tb.Sp) decrease agents, osteoarthritis severity index (OARSI) score reducers, safranin-O / fast green (SOFG) staining intensity enhancers, etc.

[0058] The agent disclosed herein can be administered directly to animals, including humans, and can also be used as a reagent. Furthermore, the agent disclosed herein can be incorporated as an ingredient in pharmaceuticals, oral compositions, food and beverages, etc. By incorporating the agent disclosed herein into an ingredient, animals, including humans, can ingest it.

[0059] The exercise-mimicking agent of this disclosure may contain a therapeutically effective amount of the above compound (1) and / or a salt thereof per unit dose. "Therapeutically effective amount" means the amount of drug effective for the treatment or prevention of a target disease. In the agent of this disclosure, the therapeutically effective amount of the above compound (1) and / or a salt thereof may be an amount effective for treatment or prevention. The therapeutically effective amount may be appropriately determined depending on the patient's symptoms, weight, age, and sex, as well as the dosage form and method of administration of the pharmaceutical composition. The exercise-mimicking agent of this disclosure may contain 0.01 to 2000 mg of the above compound (1) and / or a salt thereof per unit dose. For example, the agent of this disclosure can be added to 1,000 mL of pharmaceutical or food, preferably in dry weight amounts of 10 mg to 25,000 mg, more preferably 100 mg to 10,000 mg. Furthermore, in the agent of this disclosure, the content of compound (1) and / or its salt may be 0.01 to 90% by mass, 0.05 to 80% by mass, or 0.1 to 60% by mass.

[0060] The agents of this disclosure can be diluted with a solvent (water, organic solvent, etc.), for example. The concentration is not particularly limited. For example, low or high concentrations of the agents of this disclosure can be prepared using a 5% (v / v) aqueous solution of dimethyl sulfoxide (DMSO), for example. The concentration is not particularly limited. For example, in the case of (A) an α-adrenergic antagonist, the low concentration range is usually less than 5 mg / kg, preferably 0.01 mg / kg or more and less than 5 mg / kg, more preferably 0.1 mg / kg or more and 4 mg / kg or less, and particularly preferably 0.5 mg / kg to 3 mg / kg. The high concentration range is usually 5 mg / kg or more, preferably 5 mg / kg or more and less than 100 mg / kg, more preferably 7 mg / kg or more and 50 mg / kg or less, and particularly preferably 8 mg / kg to 25 mg / kg. Furthermore, in the case of (B) flavonoids, (C) dopamine D2 receptor antagonists, or (D) NF-κB signaling pathway inhibitors, the low concentration range is usually less than 20 mg / kg, preferably less than 10 mg / kg, preferably 0.1 mg / kg to 4 mg / kg, and more preferably 0.5 mg / kg to 3 mg / kg. The high concentration range is usually 20 mg / kg or more, preferably 20 mg / kg to less than 300 mg / kg, more preferably 22 mg / kg to 200 mg / kg, and more preferably 23 mg / kg to 100 mg / kg.

[0061] Optional Components The agents of this disclosure may contain other components in addition to compound (1) or a salt thereof. Other components include, for example, pharmaceutically acceptable carriers. "Pharmacologically acceptable carrier" means a carrier that does not inhibit the physiological activity of the active ingredient and does not exhibit substantial toxicity to the recipient. "Substantial toxicity" means that the ingredient does not exhibit toxicity to the recipient at the effective dose or dosage in which it is normally used. In the agents of this disclosure, a pharmaceutically acceptable carrier is a carrier that does not inhibit the pharmacological action of compound (1) or a salt thereof and does not exhibit substantial toxicity to the recipient. A pharmaceutically acceptable carrier includes any known pharmaceutically acceptable component that is typically considered an inactive ingredient. Pharmaceutically acceptable carriers are not particularly limited and include, for example, solvents (e.g., organic solvents such as water, DMSO, and ethanol), diluents, vehicles, excipients, flow promoters, binders, granulators, dispersants, suspending agents, wetting agents, lubricants, disintegrants, solubilizers, stabilizers, emulsifiers, and fillers. Pharmaceutically acceptable carriers may be used individually or in combination of two or more types.

[0062] The agent of this disclosure may contain other components in addition to the above components. The other components are not particularly limited, and any components commonly used in the pharmaceutical field may be used without particular restriction. Examples of other components include pharmaceutical excipients other than those mentioned above. Examples of pharmaceutical excipients include, but are not limited to, preservatives (e.g., antioxidants), chelating agents, flavoring and odor-masking agents, sweeteners, thickeners, buffering agents, and coloring agents. The agent of this disclosure may contain active ingredients other than compound (1) or its salt. Examples of active ingredients include, but are not limited to, antibiotics, anti-inflammatory agents, antipyretics, and analgesics. The other components may be used alone or in combination of two or more.

[0063] The dosage form and method of administration of the preparations disclosed herein are not particularly limited and can be any dosage form commonly used for pharmaceutical preparations. The pharmaceutical composition of this embodiment may be an oral preparation or a parenteral preparation. Examples of oral preparations include tablets, coated tablets, pills, powders, granules, capsules, syrups, fine granules, liquids, drops, emulsions, etc. Examples of parenteral preparations include injections, suppositories, nasal sprays, enteral preparations, inhalants, etc. Pharmaceutical compositions of these dosage forms can be formulated according to standard methods (for example, methods described in the Japanese Pharmacopoeia).

[0064] The administration route of the agent of this disclosure is not particularly limited and can be administered orally or parenterally. Parenteral administration includes intravenous administration, intranasal administration, subcutaneous administration, intradermal administration, intramuscular administration, intraperitoneal administration, enteral administration, etc.

[0065] The agents of this disclosure can be administered in a therapeutically effective amount of compound (1) and / or its salt. For example, the pharmaceutical composition of this embodiment can be administered in an effective single dose of 0.01 to 2000 mg of compound (1) and / or its salt per kg of body weight of the recipient. The effective dose or dosage may be 0.05 to 1500 mg / kg, 0.1 to 1000 mg / kg, 0.2 to 500 mg / kg, or 0.3 to 300 mg / kg.

[0066] The agent of this disclosure may be administered as a single dose or as a repeated dose. In the case of repeated administration, the administration interval may be appropriately determined based on the patient's symptoms, weight, age, and sex, as well as the dosage form and method of administration of the pharmaceutical composition. For example, the administration interval may be every few hours, two to three times a day, once a day, once every two to three days, once a week, once a month, once every few months, etc.

[0067] The food and beverages used are preferably foods for specified health uses, foods with nutritional function claims, health foods, functional foods, health supplements, etc. The agent disclosed herein may further contain food additives, such as sweeteners, colorants, preservatives, thickeners and stabilizers, antioxidants, bittering agents, acidulants, emulsifiers, fortifiers, processing agents, flavorings, etc. The agent disclosed herein can also be used by incorporating it into beverages such as soft drinks or other foods.

[0068] Food and beverages may have effects such as mimicking exercise effects, inhibiting fat accumulation, promoting heat production, promoting muscle differentiation, increasing skeletal muscle weight, improving exercise endurance, altering skeletal muscle fiber composition, enhancing energy metabolism, increasing basal metabolic rate, improving musculoskeletal function, and inhibiting muscle atrophy. Therefore, food and beverages may be labeled to indicate that they have these effects.

[0069] Preferred forms of food and beverages include supplements, candies, jellies, tablets, drinks, soups, noodles, rice crackers, Japanese sweets, frozen desserts, baked goods, and other food and beverage products. Preferably, packaged beverages such as fruit juices, vegetable juices, fruit and vegetable juices, tea beverages, coffee beverages, and sports drinks are included.

[0070] The agents of this disclosure may be in the form of, for example, a food composition, a supplement, or a pharmaceutical composition, and specifically, dosage forms such as liquids (including extracts and syrups), jellies, powders, granules, capsules (hard capsules, soft capsules), and tablets.

[0071] For example, when the agent of the present disclosure is added to a beverage to make a food or beverage, the agent of the present disclosure can be added in an amount of, for example, 10 mg to 25,000 mg, more preferably 100 mg to 10,000 mg, on a dry weight basis, per 1,000 mL of beverage.

[0072] When the agent of this disclosure is incorporated into food or beverages, the amount ingested can be adjusted as appropriate depending on the application, but the compounds (A) to (D) above can preferably be 10 mg to 25,000 mg per dose, more preferably 100 mg to 10,000 mg per dose, and even more preferably 100 mg to 1,000 mg per dose. The number of doses is not particularly limited, but preferably 1 to 3 times a day, and the number of doses may be increased or decreased as needed.

[0073] The number of doses is not particularly limited, but is preferably 1 to 3 times a day, and the number of doses may be increased or decreased as needed. Since the agent of this disclosure has the above-described effects, a pharmaceutical composition containing it can be used for the treatment or prevention of diseases in which the effects of the agent of this disclosure may be related to treatment or prevention. Such diseases include, for example, muscular dystrophy, low back pain, amyotrophic lateral sclerosis (ALS), spinal muscular atrophy (SMA), etc., in which effects such as muscle differentiation promotion and skeletal muscle weight increase may be related to treatment or prevention. Therefore, the agent of this disclosure can also provide a method for treating or preventing these diseases.

[0074] The target population for administration of the agents disclosed herein is not particularly limited. The target population is preferably mammals, which may be humans or other mammals. Examples of non-human mammals include non-human primates (such as monkeys, rhesus macaques, crab-eating macaques, marmosets, orangutans, and chimpanzees), rodents (such as mice, rats, hamsters, and guinea pigs), lagomorphs (such as rabbits), ungulates (such as pigs, cattle, goats, horses, and sheep), and carnivores (such as dogs and cats).

[0075] The agent disclosed herein may have potential applications in various fields, such as preventing bedridden patients due to muscle weakness in elderly people who cannot exercise, preventing muscle weakness in hospitalized patients with fractures, etc., preventing muscle weakness in outer space (low gravity), suppressing muscle weakness due to genetic diseases (e.g., muscular dystrophy), and weight loss by improving basal metabolic rate more efficiently with less exercise.

[0076] The present disclosure will be described in more detail below with reference to examples, but the technical scope of the present disclosure is not limited to these examples.

[0077] Test Example 1 (Connectivity Map) To identify compounds with potential exercise-mimicking effects, Connectivity Map (CMap), a web-based tool designed to screen compounds that modulate disease or physiological gene expression profiles, was used. Using a pharmacogenetic approach with CMap, compounds that can mimic gene expression changes observed after moderate exercise were identified. The gene expression changes due to moderate exercise were compared with the gene expression profiles of various compounds. Specifically, based on the gene expression profiles after moderate exercise, three gene lists were entered into the Connectivity Map (CMap) online tool: all genes altered by RNA-seq, hub genes identified by RNA-seq, and genes altered by RNA-seq and RRBS. RNA-seq is an analytical method that measures gene expression levels, and RRBS is an analytical method that measures epigenetic (DNA methylation) changes. By combining these two methods, important genes can be identified. CMap is a gene expression database and analysis tool developed by the Broad Institute in the United States. CMap is available via the hyperlink "http: / / scads.jfcr.or.jp / db / cmap_howto.html". These signature genes are used in CMap to query similar gene expression profiles of compound-treated cells. Table 1 and Figure 1 show the compounds or drugs ranked for connectivity score and specificity from the 2837 compounds or drugs entered into CMap.

[0078] Regarding the Connectivity Score, the Connectivity Score is a numerical representation in CMap of the similarity between the gene expression changes caused by a specific compound within a cell and known gene expression profiles. The Connectivity Score can be measured, for example, by the following procedure, but is not limited to this.

[0079] Measurement Procedure (Connectivity Score) (1) Obtaining Reference Signature: RNA is extracted from skeletal muscle (human or mouse) after aerobic / resistance exercise intervention, and RNA-seq is performed. Alignment: STAR (v2.7 or higher), Count: featureCounts, Normalization: DESeq2 (v1.3x). Significance determination: |log2 fold change| ≥ 0.58 (equivalent to 1.5 times), FDR < 0.05. The top-ranking improvement group and top-ranking decline group are sorted into a maximum of 150 genes each (including ties of the same rank), and a directional ranking list is created. (2) Compound Profile: The compound-inducible gene expression profile used for comparison is Connectivity Map compatible data (e.g., L1000 platform or equivalent). Cell line, concentration, and processing time follow the standard conditions provided by the vendor. (3) Similarity evaluation: GSEA-based connectivity (τ value or equivalent score) is used and linearly normalized to a range of -100 to +100. (4) Judgment criteria: A "positive connectivity score" means greater than +0, preferably +50 or higher, more preferably +70 or higher, and especially preferably +80 or higher. The test is performed with three or more independent replicates, and the judgment is made using the median. (5) Reproducibility: Even when data from mammals such as humans or rodents such as mice are used to create the reference signature, the judgment can be performed using the same procedure.

[0080] The score is expressed in the range of -100 to 100 and is interpreted as follows: • Positive score (0 to 100): Indicates that the gene expression changes caused by the compound are similar to known profiles. A higher score indicates a stronger similarity. • Negative score (-100 to 0): Indicates that the gene expression changes caused by the compound are inversely related to known profiles. A lower score indicates a stronger inverse correlation. • Score near 0: Suggests that the effect of the compound is not related to, or is unrelated to, known profiles. • Interpretation of a score near 0 (suggests low or no relevance). Note that (1) the gene score for aerobic exercise is an analysis based on all genes whose expression changed after exercise (analysis based on genes that changed after aerobic exercise). (2) The gene score for resistance exercise is an analysis based on all genes whose expression changed after exercise (analysis based on genes that changed after resistance exercise). (3) The hub gene score for aerobic exercise is an analysis based on hub genes (analysis based on important genes (hub genes) identified by network analysis after aerobic exercise). (4) The hub gene score for resistance exercise is an analysis based on hub genes (analysis based on hub genes identified after resistance exercise). (5) The analysis is based on genes that commonly changed in RNA-seq and RRBS analysis (analysis based on genes that commonly changed in both RNA-seq (gene expression analysis) and RRBS (DNA methylation analysis) during aerobic exercise). (6) The analysis is based on genes that commonly changed in RNA-seq and RRBS analysis (analysis based on similar common genes during resistance exercise).

[0081]

[0082] The results showed that apigenin, doxazosin, sulpiride, and BMS-345541 exhibited high connectivity scores. Apigenin showed high positive scores (98.45 and 88.77) on hub genes for aerobic and resistance exercise, making it a promising candidate as an exercise mimetic. Although not shown in Table 1, daidzein (an isoflavone) and catechin (a flavanol), both flavonoids like apigenin, also obtained scores of 50 or higher on hub genes for resistance exercise. Therefore, these compounds are also considered to be potential complementary candidates for exercise mimicry. Based on these scores and the results of previous studies, apigenin, doxazosin, sulpiride, and BMS-345541 were selected for further evaluation, and the following tests were conducted.

[0083] <Preparation> Low-concentration and high-concentration samples containing the agent disclosed herein were both prepared using a 5% (v / v) aqueous solution of dimethyl sulfoxide (DMSO) as the solvent.

[0084] Mouse 1: Male (7 weeks old) and female (7 weeks old) C57BL / 6J mice were purchased from Nippon SLC Co., Ltd. The mice were kept in plastic cages with bedding and reared under artificially controlled conditions with a temperature of 22°C ± 1°C, humidity of 55% ± 5%, and a 12-hour light / 12-hour dark cycle, with free access to water and food.

[0085] The effectiveness of the above compound in mimicking the effects of moderate exercise was investigated according to the experimental timeline of compound administration in healthy young mice shown in Figure 2.

[0086] Test Example 2 (Muscle Strength) Following the experimental timeline in Figure 2, a low-concentration sample 1 (1 mg / kg) of compound 1 (doxazosin (Selleck Chemicals, product name: S5782)) (Example 1) and a high-concentration sample 2 (10 mg / kg) of compound 1 (doxazosin (Selleck Chemicals, product name: S5782)) (Example 2) were orally administered to the above mouse 1 (healthy young mouse, C57BL / 6J mouse) for two weeks. Subsequently, the exercise-mimicking effect of the compound was investigated using the grip strength measurement method described below. In Comparative Example 1 (control), the mice were administered only a 5% DMSO aqueous solution.

[0087] Grip Strength Measurement Method: Grip strength was evaluated 24 hours after the final session of exercise or compound administration using a digital force gauge (manufactured by IMADA Corporation). During each trial, the mouse was placed on the device and gently pulled backward until the gripped hand was released. A one-minute rest period was taken between each test, and five trials were performed for each mouse. The highest value recorded in these tests was recorded as grip strength. The results are shown in Table 2 and Figure 3 below.

[0088]

[0089] The results showed that doxazosin significantly improved the grip strength of mice compared to the control group (Comparative Example 1) at both low (Example 1) and high (Example 2) concentrations, suggesting an improvement in muscle strength (Figure 3).

[0090] Test Example 3 (Muscle Strength) Following the experimental timeline in Figure 2, a low-concentration (10 mg / kg) sample 3 (Example 3) and a high-concentration (50 mg / kg) sample 4 (Example 4) of compound 2 (apigenin (MedChemExpress, product name: HY-N1201)) were orally administered to the above mouse 1 (healthy young mouse, C57BL / 6J mouse) for two weeks. Subsequently, the exercise-mimicking effect of the compound was investigated using the grip strength measurement method shown in Test Example 2. The results are shown in Table 3 and Figure 4 below.

[0091]

[0092] As a result, apigenin significantly improved the grip strength of mice compared to the control group (Comparative Example 2) at both low (Example 3) and high (Example 4) concentrations, suggesting an improvement in muscle strength (Figure 4).

[0093] Test Example 4 (Endurance Test) Following the experimental timeline in Figure 2, a low-concentration sample 5 (Example 5) containing 10 mg / kg of compound 3 (sulpiride (MedChemExpress, product name: HY-B1019)) was orally administered to mouse 1 (healthy young mouse, C57BL / 6J mouse) for two weeks. Subsequently, the following endurance test was performed to investigate the exercise-mimicking effect of the compound.

[0094] <Endurance Test> 24 hours after the final session of exercise or compound administration, the distance covered until exhaustion (all-out) was reached and further running was measured to assess endurance. Mice were accustomed to the treadmill the day before the test by running at 10 m / min for 10 minutes on a 0% gradient. During the test, the treadmill started at an initial speed of 10 m / min and a 0% gradient. The speed was increased by 2 m / min every 2 minutes, up to a maximum of 30 m / min, and continued until the mouse was exhausted. Endurance was measured by the total distance covered (meters). The results are shown in Table 4 and Figure 5 below.

[0095]

[0096] The results showed that sulpiride, at a low concentration of 10 mg / kg (Example 5), increased the total distance mice could run on a treadmill until exhaustion compared to the control group (Comparative Example 3), demonstrating its potential to enhance endurance (P = 0.058) (Figure 5).

[0097] Test Example 5 (TA Muscle Weight) Following the experimental timeline in Figure 2, a low-concentration (10 mg / kg) sample 6 (Example 6) and a high-concentration (25 mg / kg) sample 7 (Example 7) of compound 4 (BMS-345541 (MedChemExpress)) were orally administered to the above mouse 1 (healthy young mouse, C57BL / 6J mouse) for two weeks. Subsequently, the exercise-mimicking effect of the compound was investigated using the method for measuring tibialis anterior (TA) muscle weight described below.

[0098] Method for measuring TA muscle weight: Mice were anesthetized, and TA muscles were excised using sterile techniques. The excised muscles were washed with physiological saline, excess fluid was removed, and the wet weight was measured using a high-sensitivity electronic balance. The results are shown in Table 5 and Figure 6 below.

[0099]

[0100] Results: BMS-345541 increased the weight of the tibialis anterior muscle at both low (Example 6) and high (Example 7) concentrations compared to the control (Comparative Example 4) (Figure 6). These results indicate that doxazosin, apigenin, sulpiride, and BMS-345541 may act as exercise effect mimics that can effectively simulate the comprehensive molecular response to moderate exercise.

[0101] Next, following the experimental schedule shown in Figure 7, which combined apigenin or doxazosin with moderate exercise, healthy young mice (7-week-old C57BL / 6J mice) were intervened with either the compound alone, moderate exercise alone (treadmill exercise and ladder exercise), or moderate exercise plus the compound. The effects of combining these interventions with exercise on skeletal muscle function and exercise performance were then investigated.

[0102] Treadmill Exercise (Moderate Aerobic Exercise) Treadmill exercise was performed according to the following aerobic exercise protocol. A treadmill (Muromachi Machinery Co., Ltd., MK-680) was used for the aerobic exercise intervention. During the acclimatization period, all mice ran at an intensity of 8 m / min for 5 days, starting with 15 minutes and increasing by 15 minutes each day. After a 2-day rest period, the mice exercised on the treadmill at an intensity of 10-22 m / min on a 0° incline for 60 minutes. These exercise intensities were measured against maximum oxygen uptake (VO2). ２ This corresponds to approximately 70% to 95% of the maximum oxygen concentration. Various exercise protocols of different intensities were performed, and from these, the appropriate aerobic exercise intensity of 22 m / min, which maximized muscle synthesis, was determined and performed daily for four weeks.

[0103] Ladder Exercise (Moderate Resistance) Ladder exercise was performed according to the following resistance exercise protocol. Mice were introduced to the ladder device (110 cm, 80° incline) as previously described. During the habituation period, all mice climbed from bottom to top for two consecutive days. Each session consisted of 3 repetitions x 4 sets. After habituation, ladder exercise was performed at a resistance intensity ranging from 0% to 160% of the mice's body weight. From this range, the resistance intensity of 120% was determined to be the most appropriate for muscle synthesis and was performed daily for 4 weeks.

[0104] Test Example 6 (Distance Run: Moderate Exercise) Following the experimental schedule shown in Figure 7, healthy young mice (7-week-old C57BL / 6J mice) were subjected to one of the following interventions: compound only, moderate exercise only (treadmill exercise and ladder exercise), or moderate exercise plus compound. Moderate exercise consisted of treadmill exercise (moderate aerobic exercise) and ladder exercise (moderate resistance exercise), performed in the morning and afternoon, respectively. In the case of moderate exercise plus compound, the mice were administered the compound after exercise. Four weeks after the intervention, the endurance capacity tests described in Test Example 4 above were conducted on Comparative Example 5 (placebo (no compound), non-exercise (sedentary)) group, Comparative Example 6 (placebo (no compound), exercise (exercise)) group, Example 8 (compound 2 (apigenin) present, non-exercise (sedentary)) group, and Example 9 (compound 2 (apigenin) present, exercise (exercise)) group. The results are shown in Table 6 and Figure 8 below.

[0105]

[0106] The results showed that administering apigenin for four weeks increased the distance mice ran, and this increased even further when combined with exercise, suggesting an endurance-enhancing effect (Figure 8).

[0107] Test Example 7: PGC-1α Measurement Test Following the experimental schedule shown in Figure 7, healthy young mice (7-week-old C57BL / 6J mice) were intervened with either the compound alone, moderate exercise only (treadmill exercise and ladder exercise), or moderate exercise plus the compound, similar to Test Example 6. Four weeks after the intervention, PGC-1α was measured in the following groups: Comparative Example 7 (placebo (no compound), non-exercise (Sedentary)), Comparative Example 8 (placebo (no compound), exercise (Exercise)), Example 10 (compound 2 (apigenin) present, non-exercise (Sedentary)), and Example 11 (compound 2 (apigenin) present, exercise (Exercise)) using the PGC-1α measurement method described below.

[0108] Method for Measuring PGC-1α: Calf muscles were excised, treated with lysis buffer, and then centrifuged to collect the supernatant. After measuring the protein concentration, a certain amount was separated by electrophoresis and transferred to a PVDF membrane. PGC-1α was detected using a specific antibody and visualized by chemiluminescence. Representative images of Western blotting analysis of PGC-1α and GAPDH in mouse calf muscles during apigenin administration experiments are shown in Figure 9(1). The relative quantification of protein expression using the results of Western blotting analysis during apigenin administration experiments is shown in Table 7 and Figure 9(2) below (P = 0.066).

[0109]

[0110] The results showed that four weeks of apigenin administration increased PGC-1α protein levels in skeletal muscle, but combined use with exercise did not show any significant additional effect (Figure 9(2)).

[0111] Test Example 8: SDH Activity Evaluation Test Following the experimental schedule shown in Figure 7, healthy young mice (7-week-old C57BL / 6J mice) were intervened with either the compound alone, moderate exercise only (treadmill exercise and ladder exercise), or moderate exercise plus the compound, similar to Test Example 6. Four weeks after the intervention, SDH activity was measured in Comparative Example 9 (placebo (no compound), non-exercise (Sedentary)) group, Comparative Example 10 (placebo (no compound), exercise (Exercise)) group, Example 12 (compound 2 (apigenin) present, non-exercise (Sedentary)) group, and Example 13 (compound 2 (apigenin) present, exercise (Exercise)) group using the SDH activity measurement method described below.

[0112] SDH Activity Measurement Method: Succinate dehydrogenase (SDH) activity was evaluated according to a previously described protocol. Briefly, frozen muscle sections were incubated in 0.2 M sodium phosphate buffer (pH 7.4), 0.2 M sodium succinate, and 0.4 mM tetranitroblue tetrazolium in the dark at 37°C for 30 minutes. The reaction was stopped with 0.01 N hydrochloric acid for 10 seconds. The slides were then washed twice with distilled water, mounted on glycerol-gelatin, and stored in the dark for 2 days until staining intensity could be measured. All samples were treated simultaneously with the same incubation solution to ensure all samples received the same treatment. Images were taken using an Olympus BX53-33P-DPH2 (Olympus Corporation). Microscopic images of muscle were randomly selected from each section, and SDH activity was analyzed across all muscle fibers visible in each image. The average optical density (OD) of the fibers was quantified using ImageJ software (National Institutes of Health, Bethesda, MD, USA) and was considered to represent SDH activity. A representative image of SDH staining after apigenin administration (scale bar, 200 μm) is shown in Figure 10(1). The results of the SDH activity levels during the apigenin administration experiment are shown in Table 8 and Figure 10(2) below (P = 0.088).

[0113]

[0114] The results showed that administering apigenin for four weeks increased SDH activity (OD) in muscle fibers, but it did not show a significant additional effect when combined with exercise (Figure 10 (2)).

[0115] Test Example 9: Grip Strength Test Following the experimental schedule shown in Figure 7, healthy young mice (C57BL / 6J mice, 7 weeks old) were intervened with only the compound. Four weeks after the intervention, grip strength was measured in Comparative Example 11 (placebo (no compound), non-exercising (sedentary)) and Example 14 (compound 1 (doxazosin), non-exercising (sedentary)) using the grip strength measurement method described above. The results are shown in Table 9 and Figure 11 below.

[0116]

[0117] As a result, administering doxazosin for four weeks increased grip strength (Figure 11).

[0118] Test Example 10: SDH Activity Evaluation Test Following the experimental schedule shown in Figure 7, healthy young mice (C57BL / 6J mice, 7 weeks old) were intervened with either the compound alone, moderate exercise (treadmill exercise and ladder exercise) alone, or moderate exercise + the compound, and the exercise-mimicking effect was investigated. Four weeks after the intervention, the exercise-mimicking effect of the compound was investigated in Comparative Example 12 (placebo (no compound), non-exercise (Sedentary)) group, Comparative Example 13 (placebo (no compound), exercise (Exercise)) group, Example 15 (compound 1 (doxazosin) present, non-exercise (Sedentary)) group, and Example 16 (compound 1 (doxazosin) present, exercise (Exercise)) group using the SDH activity measurement method described above. A representative SDH staining image (scale bar, 200 μm) during the doxazosin administration experiment is shown in Figure 12(1). The results of the SDH activity levels during the doxazosin administration experiment are shown in Table 10 and Figure 12(2) below.

[0119]

[0120] As a result, administration of doxazosin for four weeks increased the SDH activity (OD) of muscle fibers (Figure 12 (2)).

[0121] Test Example 11: Evaluation of Type II Fiber Cross-Sectional Area Following the experimental schedule shown in Figure 7, healthy young mice (C57BL / 6J mice, 7 weeks old) were intervened with either the compound alone, moderate exercise (treadmill exercise and ladder exercise) alone, or moderate exercise + the compound to investigate the exercise effect mimicry. Four weeks after the intervention, the cross-sectional area of ​​Type II muscle fibers was calculated by the following muscle fiber analysis for Comparative Example 14 (placebo (no compound), non-exercise (Sedentary)), Comparative Example 15 (placebo (no compound), exercise (Exercise)), Example 17 (compound 1 (doxazosin) present, non-exercise (Sedentary)), and Example 18 (compound 1 (doxazosin) present, exercise (Exercise)).

[0122] Frozen sections with a thickness of 10 μm were air-dried for 20 minutes, fixed with 4% paraformaldehyde for 15 minutes, and washed twice with 0.01 M phosphate-buffered saline (PBS). The sections were blocked at room temperature for 1 hour with mouse IgG blocking reagent (Vector Laboratories, MKB-2213a) and 10% normal goat serum (Vector Laboratories, VEC S-1000). Subsequently, the tissues were exposed to mouse-on-mouse (MOM) diluent (Vector Laboratories, BMK-2202) and 5% normal goat serum at 4°C for 18 hours. The primary antibodies used were 1:1000 dilution anti-hyposkeletal myosin heavy chain mouse monoclonal antibody (Abcam, ab11083), 1:1000 dilution anti-rapid myosin heavy chain mouse monoclonal antibody (Abcam, ab51263), and 1:1000 dilution anti-dystrophin rabbit polyclonal antibody (Abcam, ab15277). For immunofluorescence, sections were also incubated at room temperature for 1 hour with 1:500 dilution of goat anti-rabbit IgG1 (H+L) highly cross-adsorbed secondary antibody, Alexa Fluor 555 (Thermo Fisher Scientific, A21429), and 1:1000 dilution of goat anti-mouse IgG1 cross-adsorbed secondary antibody, Alexa Fluor 488 (Thermo Fisher Scientific, A21121). After washing with PBS-Tx, sealing was performed using Prolong Diamond Antifade Mountant (Thermo Fisher Scientific, P36965). Immunofluorescence images of more than 300 muscle fibers in each sample and the cross-sectional area of ​​cultured muscle fibers were measured using ImageJ. Furthermore, the number of type I and type II muscle fibers was measured in the same section, and the cross-sectional area and the proportion of muscle fibers were calculated for each type of muscle fiber. Representative immunohistochemical images of type I and type II fibers of mouse gastrocnemius muscle (scale bar, 100 μm) are shown in Figure 13(1). The results of the quantification of type II muscle fiber area (S) during the doxazosin administration experiment are shown in Table 11 and Figure 13(2) below (P = 0.078).

[0123]

[0124] The results showed that four weeks of doxazosin administration increased the cross-sectional area of ​​muscle fibers (Figure 13). This suggests that doxazosin increases grip strength and the cross-sectional area of ​​type II muscle fibers, indicating that doxazosin may promote muscle fiber hypertrophy.

[0125] The results above show that apigenin alone has an effect similar to aerobic exercise, and that this effect is further amplified when combined with exercise. Conversely, doxazosin mimics the effects of resistance exercise to some extent, but its effect is not enhanced when used in combination with exercise.

[0126] Next, we investigated the exercise-mimicking effect of the above compounds on age-related diseases. Disuse muscle atrophy was chosen as an example of an age-related disease.

[0127] Test Example 12: Soleus Muscle Weight Evaluation Test Following the experimental timeline shown in Figure 14, in which apigenin or doxazosin was administered to HLU mice, the effects of apigenin and doxazosin on bone and skeletal muscle were investigated over a four-week period using a hindlimb unloading (HLU) model of mice.

[0128] Hindlimb Weight-Bearing in Mice Hindlimb weight-bearing was performed on 7-week-old male mice as follows: In short, the mice were anesthetized with 2% inhaled isoflurane and subcutaneously injected with 0.02 mg / kg of buprenorphine to relieve pain. A sterile steel wire was inserted into the intervertebral space of the tail, and the steel wire was formed into a ring and then suspended. The tail ring was attached with a rope to a track suspended from the ceiling, allowing the animals to move freely throughout the cage. The head tilt angle was monitored daily throughout the experiment and maintained at approximately 30° so that 50% of the body weight was supported by the forelimbs.

[0129] Compound 2 (apigenin) (Example 19) and Compound 1 (doxazosin) (Example 20) were administered to hindlimb unweight-bearing (HLU) model mice, respectively. Comparative Example 16 consisted of mice that did not undergo hindlimb unweight-bearing and were not administered the compounds (control group), while Comparative Example 17 consisted of mice that underwent hindlimb unweight-bearing but were not administered the compounds (HLU group). Subsequently, the exercise-mimicking effects of the compounds were investigated in the control group (Comparative Example 16), HLU group (Comparative Example 17), HLU + apigenin group (Example 19), and HLU + doxazosin group (Example 20) using the soleus muscle weight measurement method described below.

[0130] Method for measuring soleus muscle weight: Mice were anesthetized, and the soleus muscle was extracted using sterile techniques. The extracted muscle was lightly washed with physiological saline, excess fluid was removed with a paper towel, and the wet weight was measured using a high-sensitivity electronic balance. The results are shown in Table 12 and Figure 15 below.

[0131]

[0132] As a result, apigenin (Example 19) and doxazosin (Example 20) effectively prevented the decrease in soleus muscle weight associated with HLU (Figure 15).

[0133] Test Example 13: Evaluation Test of Type I Fiber Cross-Sectional Area As shown in Figure 14, the effects of apigenin and doxazosin on bone and skeletal muscle were investigated over a four-week period using a hindlimb unweight-bearing (HLU) model of mice. Compound 2 (apigenin) (Example 21) and Compound 1 (doxazosin) (Example 22) were administered to mice in the hindlimb unweight-bearing (HLU) model, respectively. Comparative Example 18 was a control group of mice that did not undergo hindlimb unweight-bearing and were not administered the compounds, while Comparative Example 19 was a HLU group of mice that underwent hindlimb unweight-bearing but were not administered the compounds. Subsequently, the cross-sectional area of ​​type I muscle fibers was calculated for the control group (Comparative Example 18), HLU group (Comparative Example 19), HLU + apigenin group (Example 21), and HLU + doxazosin group (Example 22) using the muscle fiber analysis described in Test Example 11. Photographs of immunohistochemical staining of the gastrocnemius muscle in the HLU experiment are shown in Figure 16(1). The results for the cross-sectional area of ​​type I fibers are shown in Table 13 and Figure 16(2) below (P = 0.069).

[0134]

[0135] Results: HLU did not affect the overall changes in muscle fibers (Comparative Example 19), but in doxazosin-treated mice (Example 22), the cross-sectional area of ​​type I muscle fibers tended to be larger than in the HLU-only group (Comparative Example 19) (Figure 16(2)).

[0136] Test Example 14: Evaluation Test of Bone Volume / Tissue Volume Ratio (BV / TV), Cancellous Bone Number (Tb.N), and Cancellous Bone Spacing (Tb.Sp) As shown in Figure 14, the effects of apigenin and doxazosin on bone and skeletal muscle were investigated over a four-week period using a hindlimb unweight-bearing (HLU) model of mice. Compound 2 (apigenin) (Examples 23, 25, 27) and Compound 1 (doxazosin) (Examples 24, 26, 28) were administered to mice in the hindlimb unweight-bearing (HLU) model. Comparative Examples 20, 22, and 24 were mice (controls) that did not undergo hindlimb unweight-bearing and were not administered the compounds, while Comparative Examples 21, 23, and 25 were mice (HLU) that underwent hindlimb unweight-bearing but were not administered the compounds. Subsequently, BV / TV, Tb.N, and Tb.Sp were calculated for the control group (Comparative Examples 20, 22, 24), HLU group (Comparative Examples 21, 23, 25), HLU + apigenin group (Examples 23, 25, 27), and HLU + doxazosin group (Examples 24, 26, 28) by the following μCT analysis.

[0137] After the μCT analysis experiment period, the tibias of HLU target mice were collected and scanned using a micro 3D X-ray CT system (Rigaku Corporation, R_mCT2) (operating with isotropic voxel resolution of 20 μm, voltage of 90 kV, current of 160 μA, and scan time of 180 seconds). To evaluate the changes in trabecular and cortical bone of the tibial diaphysis, 3D image reconstruction and data processing were performed using TRI / 3D-BON software (Ratoc). A region of interest 2 mm in length from the proximal tibial diaphysis directly below the growth plate was manually selected to analyze the microstructure of trabecular and cortical bone. In the trabecular bone, the concentration was 650 mg HA / cm². ３ In cortical bone, 750 mg HA / cm² ３The tissues were automatically separated using software with a threshold value. The cancellous bone structure was characterized by measuring BV / TV, Tb.Th, Tb.N, and Tb.Sp. μCT images (scale bar, 100 μm) from the HLU experiment are shown in Figure 17(1). The bone volume / tissue volume ratio (BV / TV) from the HLU experiment is shown in Table 14 and Figure 17(2). The cancellous bone number (Tb.N) is shown in Table 15 and Figure 17(3). The cancellous bone spacing (Tb.Sp) is shown in Table 16 and Figure 17(4).

[0138]

[0139]

[0140]

[0141] Results: Disuse bone atrophy caused by HLU was evident from the decrease in bone mass / tissue mass (BV / TV) ratio and cancellous bone number (Tb.N), and the increase in cancellous bone spacing (Tb.Sp) (Figure 17 (2), (3), and (4)). On the other hand, in mice administered doxazosin, BV / TV and Tb.N increased, but Tb.Sp decreased. This indicates that doxazosin can counteract the deterioration of cancellous bone caused by HLU.

[0142] Next, the exercise-mimicking effects of the above compounds in traumatic osteoarthritis of the knee induced by medial meniscus instability were investigated. Test Example 15: OARSI score evaluation test Following the experimental timeline for apigenin or doxazosin administration to traumatic osteoarthritis of the knee shown in Figure 18, the effects of apigenin and doxazosin on bone and skeletal muscle were investigated over four weeks using a mouse model of medial meniscus instability (DMM).

[0143] Medial meniscal destabilization osteoarthritis in mice was induced by medial meniscal destabilization (DMM) surgery in 7-week-old male mice under anesthesia. The surgery involved incising the medial capsule of the right knee and cutting the anteromedial meniscus tibiofibular ligament; the left knee joint was left untouched. Control mice underwent sham surgery on their right knee joint.

[0144] Doxazosin (Example 29) was administered to mice with medial meniscus destabilization (DMM). Comparative Example 26 consisted of mice (Sham group) that underwent a sham surgery to expose the medial meniscus but did not destabilize it, and were not administered the compound. Comparative Example 27 consisted of mice (DMM group) that underwent a medial meniscus destabilization (DMM) surgery but were not administered the compound. Subsequently, the exercise-mimicking effect of the compound was examined in the Sham group (Comparative Example 26), the DMM group (Comparative Example 27), and the DMM + doxazosin group (Example 29) using the OARSI score measurement method described below.

[0145] OARSI Score Measurement Method: Knee joint specimens were fixed with 4% paraformaldehyde, decalcified with 10% EDTA (ethylenediaminetetraacetic acid) / PBS (phosphate-buffered saline) treated with diethyl pyrocarbonate, and embedded in Super Cryoembedding Medium (SCEM, Leica) for the Kawamoto method. 5 μm serial sections were collected and stained with safranin-O / fast green. Figure 19 shows photographs of the safranin-O / fast green (SOFG) stained images (scale bar, 1 mm). Histological changes in articular cartilage in each group were described using the OARSI (Osteoarthritis Research Institute) score. The results are shown in Table 17 and Figure 20(1) below.

[0146]

[0147] As a result, the OARSI score, which indicates the severity of osteoarthritis, was significantly higher in DMM mice (Comparative Example 27), but there was no statistically significant difference between doxazosin-treated mice (Example 29) and imitation mice (Comparative Example 26) (Figure 20(1)).

[0148] Test Example 16: SOFG Staining Intensity Evaluation Test As shown in Figure 18, the effects of doxazosin were investigated using DMM mice over a period of four weeks. DMM mice were administered doxazosin (Example 30). Comparative Example 28 consisted of mice (Sham group) that underwent a sham surgery in which the medial meniscus was exposed but not destabilized, and were not administered the compound. Comparative Example 29 consisted of mice (DMM group) that underwent DMM surgery but were not administered the compound. Subsequently, the SOFG staining intensity was measured for Comparative Example 28, Comparative Example 29, and the DMM + doxazosin group (Example 30) using the SOFG staining intensity measurement method described below.

[0149] SOFG Staining Intensity Measurement Method: Knee joint specimens were fixed with 4% paraformaldehyde, decalcified with 10% EDTA / PBS treated with diethyl pyrocarbonate, and embedded in Super Cryoembedding Medium (Leica). 5 μm serial sections were collected and stained with safranin-O / fast green. The safranin-O / fast green staining intensity of articular cartilage in each group was measured by converting the microscopic images to grayscale and determining the resulting concentration. The results are shown in Table 18 and Figure 20(2) below.

[0150]

[0151] Results: SOFG staining intensity was observed in the DMM + doxazosin group (Example 30) (Figure 20(2)), which reflects that the proteoglycan content in the articular cartilage of doxazosin-treated DMM mice (Example 30) was higher than that of DMM mice (Comparative Example 29). These results indicate that doxazosin not only prevents proteoglycan loss in traumatic osteoarthritis of the knee but also suppresses the progression of the disease.

[0152] The results above show that the exercise-mimicking agents of this disclosure, including doxazosin, apigenin, sulpiride, or BMS-345541, not only promote endurance and muscle strength but also have beneficial effects on bones and joints. As described above, the exercise-mimicking agents of this disclosure were first identified using multi-omics analysis to identify the "pattern of gene changes that comprehensively occur in the body due to exercise (exercise signature)," and then found that at least one compound selected from the group consisting of (A) α-adrenergic antagonists, (B) flavonoids, (C) dopamine D2 receptor antagonists, and (D) NF-κB signaling pathway inhibitors of this disclosure best mimics that pattern. Such effects have not been described or suggested in any prior literature.

[0153] This application is accompanied by a priority claim based on Japanese Patent Application No. 2025-007925, filed on January 20, 2025. Japanese Patent Application No. 2025-007925 is incorporated herein by reference.

Claims

1. An exercise effect mimic containing at least one compound selected from the group consisting of (A) an α-adrenergic antagonist, (B) a flavonoid, (C) a dopamine D2 receptor antagonist, and (D) an NF-κB signaling pathway inhibitor.

2. The exercise effect mimicry agent according to claim 1, wherein the exercise effect mimicry agent contains as an active ingredient a compound that mimics an exercise reference signature, which is a change in the gene expression pattern that occurs in a living organism due to exercise.

3. The exercise effect mimicry agent according to claim 2, wherein the active ingredient is a compound that shows a connectivity score of 50 or more with respect to an exercise reference signature, which is a change in the gene expression pattern that occurs in vivo due to the exercise, based on CMap (Connectivity Map) analysis.

4. The exercise effect mimicry agent according to claim 2, wherein the exercise reference signature, which is a change in the gene expression pattern that occurs in vivo due to the exercise, is determined by multi-omics analysis including RNA sequencing analysis.

5. An exercise effect mimicking agent according to claim 1, which mimics the effects of aerobic exercise and / or resistance exercise (muscle strengthening exercise).

6. The exercise effect mimicry agent according to claim 1, wherein the exercise effect mimicry agent is the (A) α-adrenergic antagonist, and the (A) α-adrenergic antagonist is at least one compound selected from the group consisting of doxazosin, prazosin, terazosin, tamsulosin, silodosin, alfuzosin, naphtopidil, bunazosin, indramin, urapidil, phentolamine, trazoline, phenoxybenzamine, and dibenamine.

7. The exercise effect mimicry agent according to claim 1, wherein the exercise effect mimicry agent is doxazosin and mimics the effects of resistance exercise.

8. The exercise effect mimicry agent according to claim 1, wherein the exercise effect mimicry agent is the (B) flavonoid, and the (B) flavonoid contains flavon as an active ingredient.

9. The exercise effect mimicry agent according to claim 8, wherein the flavonoid is apigenin and mimics the effects of aerobic exercise.

10. The exercise effect mimicry agent according to claim 1, wherein the exercise effect mimicry agent is the (C) dopamine D2 receptor antagonist, and the (C) dopamine D2 receptor antagonist is at least one compound selected from the group consisting of sulpiride, amisulpiride, levosulpiride, tiapride, metoclopramide, rimosapride, mosapride, and nemonapride.

11. The exercise effect mimic agent is the (D) NF-κB signaling pathway inhibitor, and the (D) NF-κB signaling pathway inhibitor is BMS-345541 (CAS number: 445430-58-0), Bay11-7082 (CAS number: 19542-67-7), IMD-0354 (CAS number: 978-62-1), IMD-1041 (CAS number: 10736) The exercise effect mimicry agent according to claim 1, which is at least one selected from the group consisting of 66-73-5), IMD-2560, TPCA1 (CAS number: 507475-17-4), BOT-64 (CAS number: 113760-29-5), SC-514 (CAS number: 354812-17-2), IKK-16 (CAS number: 1186195-62-9), ertiprotafib (CAS number: 251303-04-5), and Bay65-1942 (HCl salt) (CAS number: 600734-06-3).

12. A skeletal muscle function improving agent comprising the exercise effect mimicking agent described in claim 1.

13. An energy metabolism enhancer comprising the exercise effect mimicking agent described in claim 1.

14. A chondrocyte proliferation or differentiation inducer comprising the exercise effect mimicking agent described in claim 1.

15. A gene expression promoter comprising the exercise effect mimicry agent described in claim 1, wherein the gene whose expression is promoted is a gene that improves mitochondrial function in muscle tissue.

16. The gene expression promoter according to claim 15, wherein the gene that enhances mitochondrial function in the muscle tissue is PGC-1α.

17. An oral composition or food / beverage composition comprising the exercise effect mimic agent described in claim 1.

18. A pharmaceutical composition comprising the exercise effect mimic agent described in claim 1.

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