KR20220072708A - Novel derivatives of alverine and use thereof - Google Patents

Abstract

The present invention relates to novel alberine derivatives and uses thereof. When myoblasts are treated with the alberine derivative of the present invention or a pharmaceutically acceptable salt thereof, differentiation into myotube cells is promoted. Therefore, the alberine derivative or a pharmaceutically acceptable salt thereof according to the present invention can be usefully used for promoting differentiation of myoblasts and preventing or treating diseases related to muscle weakness.

Description

Novel alberine derivatives and uses thereof

The present invention relates to novel alberine derivatives and uses thereof.

Diseases that weaken muscle strength include sarcopenia, which progresses with aging, and muscular dystrophy, a degenerative myopathy in which muscle fibers are necrotic due to genetic mutations.

Sarcopenia is a symptom of a decrease in muscle strength due to a decrease in muscle mass during aging. Sarcopenia not only decreases muscle mass, but also changes the type of muscle fibers. Such sarcopenia causes senility and functional impairment in the elderly. In addition, muscular dystrophy leads to necrosis and degeneration of muscle fibers due to deficiency or mutation of muscle-related genes, resulting in disability or death. In addition, cachexia is a muscle weakness disease that occurs frequently in cancer patients. In cancer patients, due to an increase in metabolism, the decomposition and metabolic reaction of skeletal muscle is excessive, resulting in a decrease in muscle mass and a decrease in muscle strength. Since this metabolic imbalance eventually reaches a stage where it cannot be recovered even by ingesting food, active treatment such as drug therapy is required.

Sarcopenia treatment drugs under development have been reported to have very limited efficacy in clinical trials, so there is no clear drug candidate for sarcopenia until now (Ju Yeon Kwak, Ki-Sun Kwon, Ann Geriatr Med Res . 98 -104, 2019). Therefore, only an auxiliary treatment strategy through exercise training and nutritional supplementation is known as a safe and reliable method to have a positive effect. Exercise training can exert beneficial effects on skeletal muscle through various mechanisms, including mTORC1 activation, reduction of oxidative stress, inhibition of inflammation, and increased mitochondrial production (Jiayu Yin et al. , Theranostics , 4019-4029, 2019). Although exercise training and nutritional supplementation show improvement in strength, muscle function, and muscle mass, their efficacy is limited and it is difficult to apply to the elderly who cannot exercise due to physical disabilities or whose nutrient absorption rate is low.

On the other hand, sarcopenia is a multifactorial disease with various causes, and there is no unified diagnostic standard because there is a large difference in muscle mass according to characteristics such as race, age, and sex. Recently, the 2019 EWGSOP further classified sarcopenia into three groups based on muscle mass, strength and physical function; probable-sarcopenia, confirmed sarcopenia, severe sarcopenia (AJ Cruz-Jentoft et al. , Age Aging , 601, 2019). Although the molecular and biological mechanisms that cause sarcopenia have not been clearly elucidated, mitochondrial synthesis disorders, oxidative stress, inflammation, and reduction of stem cells are thought to be the causes of sarcopenia (Jessica Hiu-tung Lo et al ., J Orthop Translat , 28-52, 2020).

Satellite cells are stem cells that exist in adult muscle and are necessary for skeletal muscle regeneration after injury. The number of satellite cells in mouse muscle appears to decrease during aging. In humans, there is a decrease in satellite cells in type II muscle. This decrease in satellite cells may contribute to the development of sarcopenia and is associated with poor muscle regeneration in aging animals (Carlson et al. , J Gerontol , B224-233, 2001).

So far, studies on the improvement of skeletal muscle hypertrophy and regeneration ability have been focused on the regulation of differentiation from myoblasts by myogenic regulatory factors (MRFs) such as MyoD, Mef2, and myogenin (Sabourin LA and Rudnicki MA., Clin Genet . 2000 Jan; 57(1):16-25.). Since the improvement of myoblast differentiation and regeneration ability by drug therapy causes muscle hypertrophy or improves muscle function, many attempts have been made to solve the decrease in muscle function due to aging by using it (Francesca Riuzzi et al. , J Cachexia) . Sarcopenia Muscle , 1255-1268, 2018, Mary F. O'Leary, Sci Rep. 12991, 2017, Jin-A Kim and Seong Min Kim, BMC Complement Altern Med , 287, 2019). That is, a method for improving the function of satellite cells and a method for increasing the pool may be a strategy for suppressing the occurrence of sarcopenia and improving the muscle regeneration ability. Accordingly, it is necessary to develop a substance that can promote the differentiation of myoblasts.

Ju Yeon Kwak, Ki-Sun Kwon, Ann Geriatr Med Res. 98-104, 2019 Jiayu Yin, Xiang Lu, Zhiyuan Qian, Weiting Xu and Xiang Zhou, Theranostics, 4019-4029, 2019 AJ Cruz-Jentoft, J.P. Baeyens, JM Bauer, Y. Boirie, T. Cederholm, F. Landi, Age Aging, 601, 2019 Jessica Hiu-tung Lo, Kin PongU, TszlamYiu, J Orthop Translat, 28-52, 2020 Carlson et al., J Gerontol, B224-233, 2001 Sabourin LA, Rudnicki MA., Clin Genet. 2000 Jan; 57(1):16-25. Francesca Riuzzi, Guglielmo Sorci, Cataldo Arcuri, J Cachexia Sarcopenia Muscle, 1255-1268, 2018, Mary F. O'Leary, Sci Rep. 12991, 2017, Jin-A Kim, Seong Min Kim, BMC Complement Altern Med, 287, 2019

Accordingly, the present inventors have endeavored to develop a substance capable of increasing muscle mass and effectively recovering muscle function by promoting the differentiation of myoblasts. As a result, the present invention was completed by confirming that an alberine derivative or a pharmaceutically acceptable salt thereof promotes differentiation of myoblasts and restores muscle strength.

In order to solve the above problems, one aspect of the present invention provides a compound represented by Formula 1 or a pharmaceutically acceptable salt thereof.

Another aspect of the present invention provides a pharmaceutical composition for preventing or treating muscle weakness-related diseases comprising the compound or a pharmaceutically acceptable salt thereof as an active ingredient.

Another aspect of the present invention provides a composition for promoting differentiation of myoblasts in vitro comprising the compound or a pharmaceutically acceptable salt thereof as an active ingredient.

Another aspect of the present invention provides a method for promoting differentiation of myoblasts, comprising treating the compound or a pharmaceutically acceptable salt thereof in vitro.

Another aspect of the present invention provides a method for preparing myotube cells, comprising the step of differentiating the myoblasts by treating the compound or a pharmaceutically acceptable salt thereof in vitro.

Another aspect of the present invention provides a food composition for preventing or improving muscle weakness-related diseases, comprising the compound or a pharmaceutically acceptable salt thereof.

Another aspect of the present invention provides a composition for strengthening muscles comprising the compound or a pharmaceutically acceptable salt thereof as an active ingredient.

When myoblasts are treated with the alberine derivative of the present invention or a pharmaceutically acceptable salt thereof, differentiation into myotube cells is promoted. Therefore, the alberine derivative according to the present invention or a pharmaceutically acceptable salt thereof can be usefully used for promoting differentiation of myocytes and preventing or treating diseases related to muscle weakness.

1 is a photograph taken by fluorescence staining of myotube cells differentiated by treating myoblasts with DMSO, insulin or alberine citrate.
2 is a graph showing the area of eMyHC fluorescence staining of myotube cells differentiated by treating myoblasts with DMSO, insulin or alberine citrate.
3 is a schematic diagram of an experiment schedule using animals to confirm the muscle strength improvement effect of alberine citrate.
Figure 4 is a graph showing the measured value of the grip force of the mouse according to the administered concentration of alberine citrate.
5 is a diagram showing the running time of mice in the normal group and the experimental group.
Figure 6 is a graph showing the time the mice of the normal group and the experimental group were hanging on the rotarod (Rotor-rod).
7 is a graph showing the muscle mass of the splenic muscle (GA) and tibialis anterior muscle (TA) obtained from mice in the normal group and the experimental group.
8 is a schematic diagram of an experiment schedule using sarcopenia-induced animals to confirm the muscle strength improvement effect of alberine citrate.
9 is a graph showing the running time of mice in a normal group, a control group and an experimental group.
10 is a graph showing the muscle mass of the splenic muscle (GA) and tibialis anterior muscle (TA) obtained from mice in a normal group, a control group and an experimental group.
11 is a photograph taken by fluorescence staining of myotube cells differentiated by treating myoblasts with DMSO, insulin, alberine citrate, or 4-hydroxy alverine (4HA).
12 is a graph showing the area of eMyHC fluorescence staining of myotube cells differentiated by treating myoblasts with DMSO, insulin, alberine citrate, or 4-hydroxy alberine (4HA).
13 is a photograph taken by fluorescently staining myotube cells inducing muscle atrophy using alberine citrate (AC) or 4-hydroxy alberine (4HA) as a cancer cell culture medium (CM).
14 is a graph showing the diameter of myotube cells inducing muscle atrophy with alberine citrate (AC) or 4-hydroxy alberine (4HA) as a cancer cell culture medium (CM).
15 is a photograph taken by fluorescence staining of myotube cells differentiated by treating myoblasts with DMSO, alberine, and 27 kinds of alberine derivatives, respectively.
16 is a graph showing the area of myotube cells differentiated by treating myoblasts with DMSO, alberine, and 27 alberine derivatives, respectively.

Hereinafter, the present invention will be described in detail.

One aspect of the present invention provides a compound represented by the following formula (1) or a pharmaceutically acceptable salt thereof:

[Formula 1]

In Formula 1,

R _{1 ,} R _{2 ,} R ₄ and R ₅ are each independently hydrogen, halogen, —CF ₃ , hydroxy, cyano, C _1-10 alkyl, C _1-10 alkoxy, C _1-4 haloalkyl, C _1-10 haloalkoxy, C _2-4 alkenyl, C _2-4 alkynyl, -NHR _a -NHC(=O)R _b , -NHC(=O)NHR _c , aminoC _1-10 alkoxy, C _1-10 alkenyloxy or C _6-10 aryl-C _1-10 alkoxy;

R ₃ is C _1-10 alkyl, C _2-12 alkenyl, C _1-4 haloalkyl, C _2-4 alkenyl or C _2-4 alkynyl;

R _a to each R _c is independently hydrogen, C _1-6 alkyl, C _1-6 alkoxy, C _1-4 haloalkyl, C _2-4 alkenyl, C _2-4 alkynyl or C _6-10 aryl;

n and m are each independently an integer of 1 to 10,

with the proviso that when n and m are 2, R ₃ is ethyl and R _{1 ,} R ₄ and R ₅ are all hydrogen, R ₂ is neither hydrogen nor hydroxy.

As used herein, the term “halo” or “halogen” means F, Cl, Br or I unless otherwise specified.

The term “alkyl,” unless otherwise specified, refers to a linear or branched saturated hydrocarbon moiety. For example, "C _1-10 alkyl" refers to alkyl having a backbone of 1 to 10 carbons. Specifically, C _1-10 alkyl is methyl, ethyl, n-propyl, i-propyl, n-butyl, i-butyl, t-butyl, n-pentyl, i-pentyl, t-pentyl, sec-pentyl, neopentyl , hexyl, heptyl, octyl, nonyl, decyl, and the like.

The term "alkoxy", unless otherwise specified, refers to a group having the formula -O-alkyl, wherein the alkyl group as defined above is attached to the parent compound through an oxygen atom. The alkyl portion of the alkoxy group has 1 to 20 carbon atoms (ie, C ₁ -C ₂₀ alkoxy), 1 to 12 carbon atoms (ie, C ₁ -C ₁₂ alkoxy), or 1 to 6 carbon atoms (ie, C 1 -C 12 alkoxy). C ₁ -C ₆ alkoxy). Examples of suitable alkoxy groups include methoxy (-O-CH ₃ or -OMe), ethoxy (-OCH ₂ CH ₃ or -OEt), t-butoxy (-OC(CH ₃ ) ₃ or -O-tBu) etc.

The term “haloalkyl” refers to an alkyl substituted with one or more halogens. Specifically, haloalkyl may be an alkyl in which two or more halogens of the same kind are substituted or two or more kinds of halogens are substituted.

The term “haloalkoxy” means alkoxy substituted with one or more halogens.

The term “aminoalkoxy” refers to alkoxy substituted with one or more amino groups.

The term “amino” refers to —NR ₂ , wherein each “R” is independently selected from H, alkyl, aryl, and the like, and typical amino groups include, but are not limited to, —NH ₂ , —N(CH ₃ ) ₂ , -NH(CH ₃ ), -N(CH ₂ CH ₃ ) ₂ , -NH(CH ₂ CH ₃ ), -NH (substituted or unsubstituted benzyl), -NH (substituted or unsubstituted phenyl), etc. may include

The term “alkenyl” is a hydrocarbon having at least one region of unsaturation, ie, a normal, secondary, tertiary or cyclic carbon atom having a carbon-carbon, sp ² double bond. For example, an alkenyl group has 2 to 20 carbon atoms (ie, C ₂ -C ₂₀ alkenyl), 2 to 12 carbon atoms (ie, C ₂ -C ₁₂ alkenyl), or 2 to 6 carbon atoms. (ie, C ₂ -C ₆ alkenyl). Examples of suitable alkenyl groups are ethylene or vinyl (-CH=CH ₂ ), allyl (-CH ₂ CH=CH ₂ ), cyclopentenyl (-C ₅ H ₇ ), and 5-hexenyl (-CH ₂ CH). ₂ CH ₂ CH ₂ CH=CH ₂ ), but is not limited thereto.

The term “alkenyloxy” refers to a group having the formula —O-alkenyl, wherein the alkenyl group as defined above is attached to the parent compound through an oxygen atom.

The term “alkynyl” is a hydrocarbon having at least one region of unsaturation, i.e., a normal, secondary, tertiary or cyclic carbon atom having a carbon-carbon, sp triple bond. For example, an alkynyl group may have 2 to 20 carbon atoms (ie, C ₂ -C ₂₀ alkynyl), 2 to 12 carbon atoms (ie, C ₂ -C ₁₂ alkynyl), or 2 to 6 carbon atoms. (ie, C ₂ -C ₆ alkynyl). Examples of suitable alkynyl groups include, but are not limited to, acetylenic (-C≡H), propargyl (-CH ₂ C≡H), and the like.

The term "aryl" means an aromatic hydrocarbon radical derived by the removal of one hydrogen atom from six carbon atoms of a parent aromatic ring system. For example, an aryl group can have 6 to 20 carbon atoms, 6 to 14 carbon atoms, or 6 to 12 carbon atoms.

The term “substitution” refers to the replacement of a hydrogen atom in a molecular structure with a substituent such that the compound is chemically stable from such substitution without exceeding the valence on the designated atom. For example, "group A is substituted with substituent B" means that a hydrogen atom bonded to an atom such as carbon constituting the backbone of group A is replaced with substituent B, so that group A and substituent B form a covalent bond can do.

In one embodiment of the compound represented by Formula 1, alverine and 4-hydroxy alverine may be excluded.

Specifically, the alberine has a structure as shown in Formula 2 below.

[Formula 2]

In this case, in the alberine, in Formula 1, n and m are 2, R ₃ is ethyl, R ₁ , R ₂ , This corresponds to the case where both R ₄ and R ₅ are hydrogen.

In addition, the 4-hydroxy alberine has a structure as shown in Formula 3 below.

[Formula 3]

In this case, the 4-hydroxy alberine corresponds to a case in which n and m are 2 in Formula 1, R ₃ is ethyl, 3 of R ₁ , R ₂ , R ₄ and R ₅ are hydrogen, and 1 is hydroxy. Specifically, when 3 of R ₁ , R ₂ , R ₄ and R ₅ are hydrogen and 1 hydroxy, i) when R ₂ , R ₄ and R ₅ are all hydrogen and R ₁ is hydroxy, ii) when R ₁ , R ₄ and R ₅ are all hydrogen and R ₂ is hydroxy iii) when R ₁ , R ₂ and R ₅ are all hydrogen and R ₄ is hydroxy and iv) R ₁ , R ₂ and R ₄ may both be hydrogen and R ₅ may be hydroxy.

According to one embodiment, in Formula 1, R ₁ , R ₂ , R ₄ and R ₅ are each independently hydrogen, halogen, hydroxy, C _1-10 alkyl, C _1-10 alkoxy, C _1-10 haloalkoxy, -NHR _a -NHC(=O)R _b , -NHC (=O)NHR _c , aminoC _1-10 alkoxy, C _1-10 alkenyloxy or C _6-10 aryl-C _1-10 alkoxy, R ₃ is C _1-10 alkyl or C _2-12 alkenyl, R _a to each R _c is independently hydrogen, C _1-6 alkyl, C _1-6 alkoxy or C _6-10 aryl; n and m may each independently be an integer of 1 to 7.

According to another embodiment, in Formula 1, R ₁ and R ₂ are each independently hydrogen, halogen, hydroxy, C _1-10 alkyl, C _1-10 alkoxy, C _1-10 haloalkoxy, -NHR _{a ,} aminoC _1-10 alkoxy, C _1-10 alkenyloxy or C _6-10 aryl-C _1-10 alkoxy, R ₃ is C _1-10 alkyl, R ₄ and R ₅ are each independently; hydrogen, hydroxy, -NHR _a , -NHC(=O)R _b or -NHC(=O)NHR _c , and R _a to each R _c is independently hydrogen, C _1-4 alkyl, C _1-6 alkoxy or C _6-10 aryl; n and m may each independently be an integer of 1 to 5.

According to another embodiment, in Formula 1, R ₁ and R ₂ are each independently hydrogen, halogen, hydroxy, C _1-6 alkyl, C _1-6 alkoxy, C _1-10 haloalkoxy, — NHR _a , aminoC _1-10 alkoxy, C _1-10 alkenyloxy or C _6-10 aryl-C _1-6 alkoxy, R ₃ is C _1-6 alkyl, R ₄ is hydrogen, halogen, hydroxy , -NHR _a , -NHC(=O)R _b or -NHC(=O)NHR _c , R ₅ is hydrogen; R _a is hydrogen or C _1-6 alkoxy; R _b is hydrogen or C _1-6 alkyl; R _c is hydrogen or C _6-10 aryl; n and m may each independently be an integer of 1 to 3.

According to specific examples, specific examples of the compound represented by Formula 1 are as follows:

1) 4-(3-ethyl(3-phenylpropyl)amino)propyl)aniline;

2) N-ethyl-3-(4-methoxyphenyl)-N-(3-phenylpropyl)propan-1-amine;

3) N-ethyl-3-phenyl-N-(3-(p-tolyl)propyl)propan-1-amine;

4) 3-(3-(ethyl(3-phenylpropyl)amino)propyl)phenol;

5) N-methyl-3-phenyl-N-(3-phenylpropyl)propan-1-amine;

6) 4,4'-((ethylazanediyl)bis(propane-3,1-diyl))diphenol;

7) 4-(3-(methyl(3-phenylpropyl)amino)propyl)phenol;

8) 4-(3-((3-(4-(benzyloxy)phenyl)propyl)(ethyl)amino)propyl)aniline;

9) N-(4-(3-((3-)4-(benzyloxy)phenyl)propyl)(ethyl)amino)propyl)phenyl)acetamide;

10) 4-(3-(ethyl(phenethyl)amino)propyl)phenol;

11) 4-(3-(ethyl(3-phenylpropyl)amino)propyl)-2-fluorophenol;

12) 2-bromo-4-(3-(ethyl(3-phenylpropyl)amino)propyl)phenol;

13) 1-(4-(3-((3-(4-(benzyloxy)phenyl)propyl)(ethyl)amino)propyl)phenyl)-3-phenylurea;

14) ethyl(4-phenylbutyl)(3-phenylpropyl)amine;

15) ethylbis(4-phenylbutyl)amine;

16) (4-phenylbutyl)(3-phenylpropyl)propylamine;

17) 3-(4-butoxyphenyl)-N-ethyl-N-(3-phenylpropyl)propan-1-amine;

18) 3-(4-(but-3-en-1-yloxy)phenyl)-N-ethyl-N-(3-phenylpropyl)propan-1-amine;

19) 3-(4-(4-bromobutoxy)phenyl)-N-ethyl-N-(3-phenylpropyl)propan-1-amine;

20) 4-(4-(3-(ethyl(3-phenylpropyl)amino)propyl)phenoxy)butan-1-amine;

21) 4-(3-(ethyl(3-(4-methoxyphenyl)propyl)amino)propyl)-2-fluorophenol;

22) 4-(3-((3-(4-(benzyloxy)-3-fluorophenyl)propyl)(ethyl)amino)propyl)aniline;

23) N-(4-(3-((3-(4-(benzyloxy)-3-fluorophenyl)propyl)(ethyl)amino)propyl)phenyl)acetamide;

24) 3-(4-(benzyloxy)-3-fluorophenyl)-N-methyl-N-(3-phenylpropyl)propan-1-amine;

25) 4-(3-(ethyl(3-(p-tolyl)propyl)amino)propyl)-2-fluorophenol;

26) N-ethyl-3-phenyl-N-(3-(4-(3-phenylpropyl)phenyl)propyl)propan-1-amine; and

27) N-ethyl-3-phenyl-N-(3-(4-((5-phenylpentyl)oxy)phenyl)propyl)propan-1-amine.

As used herein, the term “pharmaceutically acceptable salt” refers to a salt prepared according to a method conventional in the art, and the preparation method is known to those skilled in the art. Specifically, the pharmaceutically acceptable salts include, but are not limited to, salts derived from the following pharmacologically or physiologically acceptable inorganic acids and organic acids and bases. Examples of suitable acids include hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, perchloric acid, fumaric acid, maleic acid, phosphoric acid, glycolic acid, lactic acid, salicylic acid, succinic acid, toluene-p-sulfonic acid, tartaric acid, acetic acid, citric acid, methanesulfonic acid, formic acid , benzoic acid, malonic acid, naphthalene-2-sulfonic acid, benzenesulfonic acid, and the like. Salts derived from suitable bases may include alkali metals such as sodium, or potassium, alkaline earth metals such as magnesium.

Another aspect of the present invention provides a pharmaceutical composition for preventing or treating muscle weakness-related diseases, comprising the compound represented by Formula 1 or a pharmaceutically acceptable salt thereof as an active ingredient. The compound represented by Formula 1 or a pharmaceutically acceptable salt thereof is the same as described above.

The muscle weakness-related disease refers to any disease that can occur due to muscle weakness, and is a disease in which the differentiation ability of myoblasts is reduced or weakened due to a decrease in myocytes in the body or a decrease in satellite cell activity. It means a disease for which improvement or treatment can be expected. Specifically, the muscle weakness-related disease may be sarcopenia, muscular atrophy, muscular dystrophy, or cachexia.

As used herein, the term “sarcopenia” refers to a symptom of a decrease in muscle strength due to a decrease in muscle mass during aging. As the cause of the decrease in muscle mass in sarcopenia, the decrease in the activity of satellite cells is considered to be an important cause. Satellite cells are activated by stimuli such as exercise or injury to proliferate into myoblasts, and when differentiation proceeds, they fuse with other cells to form multinucleated muscle fibers.

As used herein, the term "muscular atrophy" refers to a symptom in which the muscles of the extremities gradually atrophy in an almost symmetrical manner. It can induce progressive degeneration of motor nerve fibers and cells in the spinal cord, leading to amyotrophic lateral sclerosis (ALS) and spinal progressive muscular atrophy (SPMA).

As used herein, the term “muscular dystrophy” refers to degenerative myopathy characterized by necrosis of muscle fibers regardless of the central nervous system and peripheral nervous system. The clinical symptoms of muscular dystrophy and muscular atrophy are slightly different. Muscular dystrophy mainly occurs in childhood and muscular atrophy occurs in adolescence. Also, muscular dystrophy occurs in the proximal muscle and muscular atrophy occurs in the distal muscle. The symptoms of muscle stiffness are not present in muscular dystrophy, but are present in muscular atrophy, and although muscular dystrophy is hereditary, hereditary tendency in muscular atrophy is rare.

As used herein, the term "cachexia" refers to a high degree of general weakness that can be seen in the terminal stages of cancer, tuberculosis, hemophilia, and the like. In addition, cachexia is regarded as a kind of poisoning state caused by various organ disorders throughout the body. Symptoms of cachexia include muscle weakness, rapid emaciatedness, anemia, lethargy, and yellowing of the skin. Causes of cachexia include malignant tumors, Basedo's disease, and hypopituitarism. Bioactive substances such as tumor necrosis factor (TNF) produced by macrophages were also found to be factors that enhance cachexia.

The concentration of the compound represented by Formula 1 or a pharmaceutically acceptable salt thereof in the pharmaceutical composition may be 10 μg/ml to 180 μg/ml. Specifically, the concentration of the compound represented by Formula 1 or a pharmaceutically acceptable salt thereof is 20 μg/ml to 160 μg/ml, 40 μg/ml to 140 μg/ml or 60 μg/ml to 120 μg/ml can be

In addition, the dosage of the pharmaceutical composition may be administered based on the dosage of the active ingredient, the compound represented by Formula 1 or a pharmaceutically acceptable salt thereof. Specifically, the dosage of the compound represented by Formula 1 or a pharmaceutically acceptable salt thereof may be administered in a range of 60 mg to 240 mg per day, and if the dosage is large, it may be administered 1 to 3 times a day. . Preferably, the dosage of the compound represented by Formula 1 or a pharmaceutically acceptable salt thereof is 60 mg to 120 mg, and may be administered once or twice a day.

The pharmaceutical composition may further include a pharmaceutically acceptable carrier. The carriers are commonly used in the manufacture of pharmaceuticals, and include lactose, dextrose, sucrose, sorbitol, mannitol, starch, acacia gum, calcium phosphate, alginate, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrol money, cellulose, water, syrup, methylcellulose, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, mineral oil, and the like.

The pharmaceutical composition may further include a pharmaceutically acceptable additive selected from the group consisting of lubricants, wetting agents, sweetening agents, flavoring agents, emulsifying agents, suspending agents, preservatives, and combinations thereof.

The pharmaceutical composition may be appropriately administered to a subject according to a conventional method, administration route, and dosage used in the art. Specifically, the pharmaceutical composition may be selected at an appropriate dosage and number of administration according to methods known in the art, and the dosage and number of administrations of the pharmaceutical composition actually administered depend on the type of symptom to be prevented or treated, administration It may be appropriately determined by various factors such as route, sex, health status, diet, age and weight of the individual, and the severity of the disease.

Another aspect of the present invention provides a composition for promoting differentiation of myoblasts in vitro comprising the compound represented by Formula 1 or a pharmaceutically acceptable salt thereof as an active ingredient. The compound represented by Formula 1 or a pharmaceutically acceptable salt thereof is the same as described above.

The term "extracorporeal" used in the present invention refers to a state in which, for example, a part of a living body, such as a cell or tissue, is removed and released 'out of the living body'. Specifically, the in vitro may mean "in vitro" for testing a part of a living body under artificial conditions.

The myoblast means an undifferentiated muscle cell. The myoblasts form multinuclear myotubes through fusion of mononuclear myoblasts during differentiation. In the late stage when differentiation is almost finished, the expression of myosin heavy chain (MyHC, Myosin Heavy Chain) increases.

The composition for promoting differentiation may be a medium for differentiation of DMEM containing serum, but may be included without limitation as long as it is a medium or composition capable of promoting differentiation of myoblasts. In addition, the composition may further include additional substances necessary for cell culture or differentiation.

The concentration of the compound represented by Formula 1 or a pharmaceutically acceptable salt thereof in the composition for promoting differentiation may be 0.01 μM to 100 μM. Specifically, the concentration of the compound represented by Formula 1 or a pharmaceutically acceptable salt thereof is 0.1 μM to 50 μM, 0.1 μM to 25 μM, 0.5 μM to 25 μM, 0.1 μM to 10 μM, 0.5 μM to 10 μM, 1 μM to 10 μM, 0.1 μM to 5 μM, 0.5 μM to 5 μM, 1 μM to 5 μM, 0.1 μM to 2 μM, 0.5 μM to 2 μM or 1 μM to 2 M. In an embodiment of the present invention, C2C12 cells, which are mouse-derived myoblasts, were treated at a concentration of 1 μM.

Another aspect of the present invention provides a method for accelerating the differentiation of myoblasts, comprising treating the compound represented by Formula 1 or a pharmaceutically acceptable salt thereof to the myoblasts in vitro. The method for promoting differentiation of myoblasts may be performed in vitro or ex vivo .

Another aspect of the present invention provides a method for producing myotube cells, comprising the step of differentiating the myoblast cells by treating the compound represented by Formula 1 or a pharmaceutically acceptable salt thereof in vitro. The method for preparing the myotube cells may be performed in vitro or ex vivo .

Another aspect of the present invention provides a food composition for preventing or improving muscle weakness-related diseases, comprising the compound represented by Formula 1 or a pharmaceutically acceptable salt thereof. When the food composition is used as a food additive, it may be added as it is or used together with other foods or food ingredients, and may be appropriately used according to a conventional method.

The food composition may be used before or after the onset of muscle weakness-related disease in order to prevent or improve muscle weakness-related diseases, simultaneously with or separately from a drug for disease treatment. Specifically, the food composition is characterized in that it promotes differentiation of myoblasts. The muscle weakness-related disease is the same as described above in the pharmaceutical composition.

Another aspect of the present invention provides a composition for strengthening muscles comprising the compound represented by Formula 1 or a pharmaceutically acceptable salt thereof or a food pharmaceutically acceptable salt thereof as an active ingredient. The composition for strengthening the muscle may be used as a pharmaceutical composition or a food composition.

The strengthening of the muscle means an increase in muscle mass, strengthening of muscle recovery, and reduction of muscle fatigue. The composition for strengthening the muscle can increase the total muscle mass by increasing the muscle mass through the ability to differentiate myoblasts into muscle cells, and thus can also reduce muscle fatigue. In addition, because muscle cells can be replaced quickly, it can heal quickly for muscle damage. The composition for strengthening the muscles of the present invention may be used as a feed or feed additive.

Hereinafter, preferred examples are presented to help the understanding of the present invention. However, the following examples are only provided for easier understanding of the present invention, and the contents of the present invention are not limited by the following examples.

Experimental Example 1. Confirmation of the differentiation promoting effect of alberine citrate in myoblasts

Experimental Example 1.1. of the myoblast strain C2Cl2 culture

C2Cl2 (American Type Culture Collection, CRL-1772??) cells are myoblast cell lines obtained from C3H mice, and are used for myoblast differentiation studies. The C2C12 cells were cultured using a culture medium, and then cultured using a differentiation medium when differentiation was induced. At this time, DMEM supplemented with 10% fetal bovine serum was used as a growth media (GM, growth media), and 5% horse serum (HS) as a differentiation media (DM) was used. This added DMEM was used.

Experimental Example 1.2. Promote differentiation of myoblast line

In order to confirm the differentiation-promoting effect of alberine citrate on myoblasts, C2C12 cells were dispensed in the culture medium of Example 1.1 and cultured for 24 hours, and then DMSO, insulin or alberine citrate was added to the C2C12 cells, respectively. (Alverine citrate) was treated and differentiation was induced for 3 days. Thereafter, differentiation into myotube cells and changes in the thickness and diameter of myotube cells were observed through fluorescence staining using an antibody against myosin H chain (hereinafter, MyHC). At this time, DMSO, insulin, and alberine citrate used in the experiment were all purchased from Sigma-Aldrich. The group treated with insulin was used as a positive control.

Specifically, 5×10 ⁵ C2C12 cells were dispensed per plate in a 6-well-plate treated with the culture medium of Example 1.1, and after 24 hours, it was replaced with DMEM medium supplemented with 5% horse serum. Differentiation was induced. Thereafter, DMSO, insulin (1.72 μM) or alberine citrate (0.01 μM, 0.1 μM, 1 μM or 10 μM) were treated, respectively. After 3 days, the medium was removed, washed with a phosphate buffer solution (1X PBS), and treated with paraformaldehyde (4%) to fix the cells at room temperature for 15 minutes. Then, after washing 3 times with phosphate buffer solution (1X PBS), PBS containing 0.3% triton X-100 was treated as a permeabilization buffer, and then reacted at room temperature for 10 minutes.

After washing 3 times with phosphate buffer solution (1X PBS), it was treated with PBST (blocking buffer, PBS containing 0.5% Tween 20) containing 2% bovine serum albumin and reacted for 30 minutes to prevent non-specific Antibody binding was inhibited. After washing 3 times with a phosphate buffer solution (1X PBS), 100 μl of a primary antibody against MYH3 (SC-20641, Santa Cruz Biotechnology) diluted at a 1:500 ratio was added and reacted at room temperature for 1 hour. Then, it was washed 3 times with phosphate buffer solution (1X PBS), and 100 μl of a secondary antibody (Goat anti-Rabbit IgG-HRP) diluted at a ratio of 1:5,000 was added and reacted at room temperature for 1 hour. After 1 hour, in order to stain the cell nucleus, it was washed 3 times with a phosphate buffer solution (1X PBS), diluted with DAPI staining reagent in the blocking buffer, and then reacted at room temperature for 10 minutes. After that, it was washed 3 times with a phosphate buffer solution (1X PBS), and the washed cover glass was compared and analyzed by measuring absorbance and taking a fluorescence photograph at a wavelength of 450 nm using a fluorescence microscope. The stained area of the anti-MyHC antibody was analyzed using imageJ (Java-based image processing program).

As a result, it was confirmed that myoblasts treated with insulin and alberine citrate were more differentiated into myotubes than myoblasts treated with DMSO alone. In particular, as the treatment concentration of alberine citrate increased, the diameter of myotube cells also increased ( FIGS. 1 and 2 ).

Experimental Example 2. Confirmation of muscle strength improvement effect of alberine citrate using normal mice

In order to confirm the muscle strength improvement effect of alberine citrate, after administration of alberine citrate to mice, exercise abilities such as grip strength, running, and balance coordination were measured. After measuring exercise capacity, muscle mass was measured. Specifically, alberine citrate was administered to 10-week-old C57BL/6 male mice at 17 mg/kg/day (AC-L), 50 mg/kg/day (AC-M), or 150 mg/day for 4 weeks. 200 μl per mouse was orally administered at a concentration of kg/day (AC-H), which was set as an experimental group (FIG. 3). 10-week-old C57BL/6 male mice were purchased from Daehan Biolink.

Experimental Example 2.1. grip force measurement

Grip strength was measured using a mouse grip force meter (bioseb, USA). Specifically, the mouse was placed on the wire mesh connected to the instrument panel to observe the strength of the grip, and the strength of the mouse gripping the wire mesh was measured while grabbing the tail and dragging it downward. At this time, the measurement was repeated 5 times in succession, and the average value of the 5 measurement values was calculated.

As a result, the grip strength of the mice of the experimental group was measured to be higher than that of the mice of the normal group. Through this, it was confirmed that alberine citrate was effective in improving muscle strength (FIG. 4).

Experimental Example 2.2. measuring running ability

Running ability was measured using a self-made treadmill. Specifically, first, an electrical stimulus was allowed to flow at the starting point, causing a sense of rejection. It was acclimatized for 10 minutes at a speed of 8 m/min before measurement. After the mice in each group were placed in an isolated lane and allowed to run, the time until the mice became tired was measured. To judge whether the mouse was tired, if the time to stay outside the lane without running for more than 10 seconds passed, it was judged that the mouse was tired and the time was recorded. This experiment could not be repeated on the same mouse. Mice were placed on a trade mill and started at a speed of 8 rpm, accelerated by 2 rpm every 10 minutes, and run at a maximum speed of 20 rpm. In addition, starting with no slope, the slope was increased by 5 degrees after 30 minutes.

As a result, the running time of the mice of the experimental group increased more than that of the mice of the normal group, and in particular, the running time of the experimental group treated with alberine citrate at a concentration of 150 mg/kg/day was significantly increased ( FIG. 5 ).

Experimental Example 2.3. Measurement of balance ability

The balance control ability was measured through the Rota-Rod Test. The rotor rod device consists of 4 test zones with a shaft diameter of 3 cm and a lane width of 9 cm, and consists of 5 rotatable circular partitions with a diameter of 60 cm and a circular rod. The measurement started with a rotation speed of 10 rpm and accelerated until it reached a maximum speed of 40 rpm for 5 minutes, and the time the mouse remained in the rota rod device without hanging from the circular rod was measured. After resting for 15 minutes, the process of measuring again was repeated 3 times to calculate the average value of the measured values.

As a result, the time the mice of the experimental group hung on the round rod was increased than the time that the normal mice hung on the circular rod. The time there was also increased proportionally (FIG. 6).

Experimental Example 2.4. muscle mass measurement

To measure the muscle mass, the gastrocnemius (GA) and tibialis anterior (TA) of the hind legs of each group of mice were excised and the weight was measured. As a result, the muscle mass of the splenic and tibialis anterior muscles of the mice of the experimental group increased more than that of the mice of the normal group. In particular, as the concentration of alberine citrate treatment increased, the muscle mass of the splenic and tibialis muscles of the mice of the experimental group also increased (FIG. 7) .

Experimental Example 3. Confirmation of muscle strength improvement effect of alberine citrate using sarcopenia-induced mice

In order to confirm the muscle strength improvement effect of alberine citrate, after administration of alberine citrate to sarcopenia-induced mice, running ability was measured. After measuring running ability, muscle mass was measured. Specifically, the hind legs of 10-week-old C57BL/6 male mice were fixed using a surgical stapler (Autosuture Royal 35W stapler). After 5 days, the stapler was removed and there was a recovery period of 3 days. Then, running ability was measured. Alberine was orally administered at a concentration of 17 mg/kg/day (AC-L), 50 mg/kg/day (AC-M) or 150 mg/kg/day (AC-H) at 200 μl per mouse, This was set as the experimental group. Sarcopenia-induced mice that were not treated with anything were set as a control group ( FIG. 8 ). 10-week-old C57BL/6 male mice were purchased from Daehan Biolink.

Experimental Example 3.1. measuring running ability

Running ability was measured in the same manner as in Example 2.2. As a result, in the case of the control group mice, the running time was reduced by about 10 minutes compared to the normal group mice. On the other hand, the running time of mice in the experimental group increased in proportion to the treatment concentration of alberine citrate, and in particular, in the case of the experimental group administered orally with 150 mg/kg/day (AC-H) of alberine, the running time compared to normal mice. increased (Fig. 9).

Experimental Example 3.2. muscle mass measurement

To measure the muscle mass, the splenic spleen (GA) and tibialis anterior (TA) muscles of the hind legs of each group were excised and their weights were measured. As a result, the weight of the splenic muscle and tibialis anterior muscle was significantly reduced in the control group compared to the normal group. However, in the case of the experimental group, as the concentration of alberine citrate treatment increased, the muscle mass of the splenic and tibialis anterior muscles of the mice of the experimental group also increased ( FIG. 10 ).

Experimental Example 4. Confirmation of the differentiation promoting effect of 4-hydroxy alberine in myoblast cells

In order to confirm the differentiation-promoting effect of 4-hydroxy alberine in myoblasts, DMSO, insulin (1.72 µM), alberine citrate (0.01 µM, 0.1 µM, 1 µM or 10 µM) were used in the same manner as in Example 1. , 4-hydroxyalberine (0.01 μM, 0.1 μM, 1 μM or 10 μM) was treated, respectively, and the differentiation into myotube cells and changes in the thickness and diameter of myotube cells were observed. In this case, the alberine and 4-hydroxy alberine are shown in Table 1 below.

As a result, it was confirmed that myoblasts treated with alberine citrate or 4-hydroxy alberine were more differentiated into myotubes than myoblasts treated with DMSO or insulin ( FIGS. 11 and 12 ). 4-hydroxy alberine had a superior differentiation promoting effect than alberine at the same concentration (FIG. 12).

Experimental Example 5. Cachexia treatment effect of alberine citrate and 4-hydroxy alberine

In order to confirm the cachexia therapeutic effect of alberine citrate and 4-hydroxy alberine, an experiment was conducted to mimic the environment in which tumors were created by treating cancer cell conditioned medium (CM) as an in vitro approach. First, a cancer cell culture medium was prepared using the C26 cancer cell line. In this case, RPMI1640 medium supplemented with 10% fetal bovine serum was used as a cancer cell culture medium (GM, Growth media). The C26 cancer cell line was cultured using a 100 mm cell culture plate and the cancer cell culture medium. At this time, the medium was removed when the cell growth degree (cell confluency) reached 90%. Thereafter, after washing with a phosphate buffer solution (1X PBS), the differentiation medium prepared in the same manner as in Example 1 was treated. After 24 hours, CM (Conditioned medium), a culture medium of C26 cancer cell line, was obtained, and was used after filtration with Bottle Top Filters (Thermo, PES, 1L).

5×10 ⁵ C2C12 cells per plate were dispensed in 6-well-plates treated with the culture medium of Example 1.1, and after 24 hours, they were replaced with DMEM medium supplemented with 5% horse serum to induce differentiation. did After 4 days, the differentiated myotube cells were treated with a differentiation medium supplemented with 33% CM, and myotube cell atrophy was induced for 3 days. At this time, as the experimental group, alberine citrate (AC) and 4-hydroxy alverine (4HA) were treated at a concentration of 1 μM to evaluate the inhibitory effect of myotube cell atrophy. DMSO was used as a negative control, and ursolic acid (UA) was used as a positive control. In the same manner as in Example 1.2, the expression level of the myosin heavy chain was measured through fluorescence staining using an antibody against myosin H chain (hereinafter, MyHC).

As a result, it was confirmed that the myotube cells of the negative control were atrophied by CM. On the other hand, it was confirmed that CM-induced myotube cell atrophy was inhibited by treatment with alberine citrate or 4-hydroxy alberine ( FIGS. 13 and 14 ).

Example 1. Confirmation of the effect of promoting differentiation of myoblasts of alberine derivatives

Example 1.1. of the myoblast strain C2Cl2 culture

C2Cl2 (American Type Culture Collection, CRL-1772??) cells are myoblast cell lines obtained from C3H mice, and are used for myoblast differentiation studies. The C2C12 cells were cultured using a culture medium, and then cultured using a differentiation medium when differentiation was induced. At this time, DMEM supplemented with 10% fetal bovine serum was used as a growth media (GM, growth media), and 5% horse serum (HS) as a differentiation media (DM) was used. This added DMEM was used.

Example 1.2. Promote differentiation of myoblast line

In order to confirm the differentiation-promoting effect of the alberine derivative of myoblasts, C2C12 cells were dispensed in the culture medium of Example 1.1 and cultured for 24 hours, and then DMSO (Sigma-Aldrich), alverine (alverine, Sigma-Aldrich) or 27 kinds of alberine derivatives were treated at a concentration of 1 μM and differentiation was induced for 4 days. Thereafter, differentiation into myotube cells and changes in the thickness and diameter of myotube cells were observed through fluorescence staining using an antibody against myosin H chain (hereinafter, MyHC). At this time, 27 kinds of alberine derivatives were produced by requesting synthesis from Korea Research Institute of Bioscience and Biotechnology and JD Bioscience, and alberine and 27 kinds of alberine derivatives 1 to 27 used in the experiment are shown in Table 2 below.

Specifically, 5×10 ⁵ C2C12 cells were dispensed per plate in a 6-well-plate treated with the culture medium of Example 1.1, and after 24 hours, it was replaced with DMEM medium supplemented with 5% horse serum. Differentiation was induced. Thereafter, DMSO, alberine, or 27 kinds of alberine derivatives were treated at a concentration of 1 μM. After 4 days, the medium was removed, washed with a phosphate buffer solution (1X PBS), and treated with paraformaldehyde (4%) to fix the cells at room temperature for 15 minutes. Then, after washing 3 times with phosphate buffer solution (1X PBS), PBS containing 0.3% triton X-100 was treated as a permeabilization buffer, and then reacted at room temperature for 10 minutes.

After washing 3 times with phosphate buffer solution (1X PBS), PBST (blocking buffer, PBS containing 0.5% Tween 20) containing 2% bovine serum albumin was treated and reacted for 30 minutes to prevent non-specific Antibody binding was inhibited. After washing 3 times with phosphate buffer solution (1X PBS), 100 μl of a primary antibody against MYH3 (SC-20641, Santa Cruz Biotechnology) diluted at a 1:500 ratio was added and reacted at room temperature for 1 hour. Then, it was washed 3 times with phosphate buffer solution (1X PBS), and 100 μl of a secondary antibody (Goat anti-Rabbit IgG-HRP) diluted at a 1:5,000 ratio was added and reacted at room temperature for 1 hour. After 1 hour, in order to stain the cell nucleus, it was washed 3 times with a phosphate buffer solution (1X PBS), diluted with a DAPI staining reagent in the blocking buffer, and then reacted at room temperature for 10 minutes. Then, it was washed 3 times with a phosphate buffer solution (1X PBS), and the washed cover glass was compared and analyzed by measuring absorbance and taking a fluorescence photograph at a wavelength of 450 nm using a fluorescence microscope.

As a result, it was confirmed that all myoblasts treated with 27 kinds of alberine derivatives promoted the differentiation into myotubes similar to myoblasts treated with alberine compared to myoblasts treated with DMSO alone (Fig. 15 and Fig. 16).

Claims (13)

A compound represented by the following formula (1) or a pharmaceutically acceptable salt thereof:
[Formula 1]

In Formula 1,
R _{1 ,} R _{2 ,} R ₄ and R ₅ are each independently hydrogen, halogen, —CF ₃ , hydroxy, cyano, C _1-10 alkyl, C _1-10 alkoxy, C _1-4 haloalkyl, C _1-10 haloalkoxy, C _2-4 alkenyl, C _2-4 alkynyl, -NHR _a , -NHC(=O)R _b , -NHC(=O)NHR _c , aminoC _1-10 alkoxy, C _1-10 alkenyloxy or C _6-10 aryl-C _1-10 alkoxy;
R ₃ is C _1-10 alkyl, C _2-12 alkenyl, C _1-4 haloalkyl, C _2-4 alkenyl or C _2-4 alkynyl;
R _a to each R _c is independently hydrogen, C _1-6 alkyl, C _1-6 alkoxy, C _1-4 haloalkyl, C _2-4 alkenyl, C _2-4 alkynyl or C _6-10 aryl;
n and m are each independently an integer of 1 to 10,
with the proviso that when n and m are 2, R ₃ is ethyl and R _{1 ,} R ₄ and R ₅ are all hydrogen, R ₂ is neither hydrogen nor hydroxy. The method of claim 1,
In Formula 1,
R _{1 ,} R _{2 ,} R ₄ and R ₅ are each independently hydrogen, halogen, hydroxy, C _1-10 alkyl, C _1-10 alkoxy, C _1-10 haloalkoxy, -NHR _a -NHC (=O )R _b , —NHC(=O)NHR _c , aminoC _1-10 alkoxy, C _1-10 alkenyloxy or C _6-10 aryl-C _1-10 alkoxy;
R ₃ is C _1-10 alkyl or C _2-12 alkenyl,
R _a to each R _c is independently hydrogen, C _1-6 alkyl, C _1-6 alkoxy or C _6-10 aryl;
n and m are each independently an integer of 1 to 7, a compound or a pharmaceutically acceptable salt thereof. According to claim 1,
In Formula 1,
R ₁ and R ₂ are each independently hydrogen, halogen, hydroxy, C _1-10 alkyl, C _1-10 alkoxy, C _1-10 haloalkoxy, -NHR _a, aminoC _1-10 alkoxy, C _{1 10} alkenyloxy or C _6-10 aryl-C _1-10 alkoxy;
R ₃ is C _1-10 alkyl,
R ₄ and R ₅ are each independently hydrogen, hydroxy, C _1-10 alkoxy, -NHR _a , -NHC(=O)R _b or -NHC(=O)NHR _c ;
R _a to each R _c is independently hydrogen, C _1-4 alkyl, C _1-6 alkoxy or C _6-10 aryl;
n and m are each independently an integer of 1 to 5, a compound or a pharmaceutically acceptable salt thereof. According to claim 1,
In Formula 1,
R ₁ and R ₂ are each independently hydrogen, halogen, hydroxy, C _1-6 alkyl, C _1-6 alkoxy, C _1-6 haloalkoxy, —NHR _a , aminoC _1-10 alkoxy, C _1-10 alkenyloxy or C _6-10 aryl-C _1-6 alkoxy;
R ₃ is C _1-6 alkyl,
R ₄ is hydrogen, halogen, hydroxy, C _1-6 alkoxy, -NHR _a , -NHC(=O)R _b or -NHC(=O)NHR _c ;
R ₅ is hydrogen;
R _a is hydrogen or C _1-6 alkoxy;
R _b is hydrogen or C _1-6 alkyl;
R _c is hydrogen or C _6-10 aryl;
n and m are each independently an integer of 1 to 3, a compound or a pharmaceutically acceptable salt thereof. According to claim 1,
The compound represented by the formula (1)
1) 4-(3-ethyl(3-phenylpropyl)amino)propyl)aniline;
2) N-ethyl-3-(4-methoxyphenyl)-N-(3-phenylpropyl)propan-1-amine;
3) N-ethyl-3-phenyl-N-(3-(p-tolyl)propyl)propan-1-amine;
4) 3-(3-(ethyl(3-phenylpropyl)amino)propyl)phenol;
5) N-methyl-3-phenyl-N-(3-phenylpropyl)propan-1-amine;
6) 4,4'-((ethylazanediyl)bis(propane-3,1-diyl))diphenol;
7) 4-(3-(methyl(3-phenylpropyl)amino)propyl)phenol;
8) 4-(3-((3-(4-(benzyloxy)phenyl)propyl)(ethyl)amino)propyl)aniline;
9) N-(4-(3-((3-)4-(benzyloxy)phenyl)propyl)(ethyl)amino)propyl)phenyl)acetamide;
10) 4-(3-(ethyl(phenethyl)amino)propyl)phenol;
11) 4-(3-(ethyl(3-phenylpropyl)amino)propyl)-2-fluorophenol;
12) 2-bromo-4-(3-(ethyl(3-phenylpropyl)amino)propyl)phenol;
13) 1-(4-(3-((3-(4-(benzyloxy)phenyl)propyl)(ethyl)amino)propyl)phenyl)-3-phenylurea;
14) ethyl(4-phenylbutyl)(3-phenylpropyl)amine;
15) ethylbis(4-phenylbutyl)amine;
16) (4-phenylbutyl)(3-phenylpropyl)propylamine;
17) 3-(4-butoxyphenyl)-N-ethyl-N-(3-phenylpropyl)propan-1-amine;
18) 3-(4-(but-3-en-1-yloxy)phenyl)-N-ethyl-N-(3-phenylpropyl)propan-1-amine;
19) 3-(4-(4-bromobutoxy)phenyl)-N-ethyl-N-(3-phenylpropyl)propan-1-amine;
20) 4-(4-(3-(ethyl(3-phenylpropyl)amino)propyl)phenoxy)butan-1-amine;
21) 4-(3-(ethyl(3-(4-methoxyphenyl)propyl)amino)propyl)-2-fluorophenol;
22) 4-(3-((3-(4-(benzyloxy)-3-fluorophenyl)propyl)(ethyl)amino)propyl)aniline;
23) N-(4-(3-((3-(4-(benzyloxy)-3-fluorophenyl)propyl)(ethyl)amino)propyl)phenyl)acetamide;
24) 3-(4-(benzyloxy)-3-fluorophenyl)-N-methyl-N-(3-phenylpropyl)propan-1-amine;
25) 4-(3-(ethyl(3-(p-tolyl)propyl)amino)propyl)-2-fluorophenol;
26) N-ethyl-3-phenyl-N-(3-(4-(3-phenylpropyl)phenyl)propyl)propan-1-amine; and
27) a compound selected from the group consisting of N-ethyl-3-phenyl-N-(3-(4-((5-phenylpentyl)oxy)phenyl)propyl)propan-1-amine, or a pharmaceutically acceptable compound thereof possible salts. A pharmaceutical composition for preventing or treating muscle weakness-related diseases, comprising the compound of any one of claims 1 to 5 or a pharmaceutically acceptable salt thereof as an active ingredient. 7. The method of claim 6,
The muscle weakness-related disease is sarcopenia (sarcopenia), muscular atrophy (muscular atrophy), muscular dystrophy (muscular dystrophy) or cachexia (cachexia), muscle weakness-related disease prevention or treatment pharmaceutical composition. A composition for promoting differentiation of in vitro myoblasts comprising the compound of any one of claims 1 to 5 or a pharmaceutically acceptable salt thereof as an active ingredient. A method for promoting differentiation of myoblasts, comprising the step of treating the compound of any one of claims 1 to 5 or a pharmaceutically acceptable salt thereof in vitro. [Claim 6] A method for producing myotube cells, comprising the step of differentiating the myoblasts by treating the compound of any one of claims 1 to 5 or a pharmaceutically acceptable salt thereof in vitro. A food composition for preventing or improving muscle weakness-related diseases, comprising the compound of any one of claims 1 to 5 or a pharmaceutically acceptable salt thereof. 12. The method of claim 11,
The muscle weakness-related disease is sarcopenia, muscle atrophy (muscular atrophy), muscular dystrophy (muscular dystrophy) or cachexia (cachexia), the food composition. [Claim 6] A composition for strengthening muscle strength comprising the compound of any one of claims 1 to 5 or a pharmaceutically acceptable salt thereof as an active ingredient.

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