Composition for preventing and treating muscle diseases containing Magnolia officinalis extract as an active ingredient

Abstract

The present invention relates to a pharmaceutical composition for treating muscle diseases, comprising Magnolia officinalis extract, which contains various ingredients in addition to magnolol and honokiol, and does not interfere with anti-tumor functions, while suppressing weight loss, muscle mass loss, grip strength loss and muscle fiber damage caused by cisplatin, and promoting the restoration of damaged muscle fibers through macrophage polarization from M1 to M2. Therefore, it can be usefully used in the prevention and treatment of muscle diseases, particularly muscle diseases caused by anti-cancer drugs.

Description

[Technical field]

[0001] The present invention relates to a pharmaceutical composition for treating muscular diseases, comprising an extract of Magnolia officinalis. [Background technology]

[0002] Muscles make up about 40% of the human body, and maintaining the proper amount of muscle is essential to maintain the functional capabilities of the human body and prevent metabolic diseases. They are broadly divided into smooth muscle, cardiac muscle, and skeletal muscle, and skeletal muscle accounts for a significant portion of the human body and promotes skeletal movement. Skeletal muscle is the largest organ in the human body, accounting for 40-50% of the total body weight, and plays an important role in various metabolic functions in the body, including energy homeostasis and heat generation. From the age of 40, human muscle decreases by more than 1% every year, and by the age of 80, 50% of maximum muscle mass has been reduced, and muscle loss in old age is recognized as the most important factor in the decline of overall physical function.

[0003] Muscle diseases cause gradual impairment of walking and mobility due to weakening of skeletal muscles, making activities of daily living (ADL) difficult and making it impossible to live independently. In addition, it leads to cardiopulmonary dysfunction and other complications. Among these, cachexia is a syndrome that often accompanies chronic diseases such as cancer, tuberculosis, AIDS, and chronic obstructive pulmonary disease, and refers to a catabolic state of the body's metabolism accompanied by persistent loss of appetite and weight loss, resulting in malnutrition, metabolic imbalance, and loss of muscle and fat. Unlike other chronic disease patients, cancer patients are characterized by not only cachexia but also side effects of various anti-cancer therapies used to treat cancer (Fearon K. et al., Lancet Oncol 2011;12(5):489-495).

[0004] Cachexia occurs in 50-80% of patients with gastrointestinal and lung cancers, and the mortality rate due to cachexia is 20-30%. Cancer cachexia is characterized by muscle loss and weight loss due to increased inflammatory and metabolic catabolic responses caused by various cytokines, and occurs when muscle wasting causes weight loss of more than 5% within 12 months. These changes reduce the response rate to anticancer chemotherapy and radiation therapy, make it difficult to progress with effective anticancer treatment, and reduce the patient's quality of life and shorten survival. Muscle loss is one of the most prominent characteristics of cachexia, and is known to be due to increased protein catabolism and decreased protein production due to the overactivation of various cytokines. Cachexia is a condition that includes muscle loss (sarcopenia), and there is a lot of overlap between the two conditions. Most patients with cachexia have muscle loss (sarcopenia), but not all patients who show muscle loss show cachexia symptoms. Clinically, muscle loss (sarcopenia) can be said to be a precursor to cachexia. Inflammatory cytokines that act in cachexia affect insulin and testosterone, which regulate muscle metabolism, causing abnormalities in muscle protein synthesis (Ryu, Seung-wan, J. Clin. Nutr. 2017;9:2-6).

[0005] Sarcopenia, one of the characteristics of cachexia, occurs when the motor nerves that induce skeletal muscle contraction degenerate, causing skeletal muscle contraction to not proceed, or when the expression of proteins involved in muscle contraction in skeletal muscle is reduced or deformed, causing normal skeletal muscle contraction to not proceed, and in the long term, the motor nerves or skeletal muscle are transformed into fibrous tissue, which can be caused by a variety of causes, including aging, hormonal abnormalities, nutritional deficiencies, lack of physical activity, inflammation, and degenerative diseases, among which cancer and anti-cancer chemotherapy treatment are known to be the main causes. Currently, it is known that exercise, protein, and calorie supplementation are effective for sarcopenia, but these are not very effective for patients and elderly people, who account for the majority of sarcopenia patients, and there is a dire need for sarcopenia treatments. However, the currently used treatments for sarcopenia are still at the clinical trial level, and there are currently no drugs that have been approved by the FDA for their direct effect of improving sarcopenia and increasing muscle mass. Therefore, there are efforts to develop selective androgen receptor modulators, activin receptor antagonists, fast skeletal muscle troponin inhibitors, etc. as sarcopenia treatment agents, but they are currently at the stage of initial clinical trials. Currently, the sarcopenia treatment method mainly involves suppressing muscular atrophy caused by the degeneration or progressive mutation of muscle cells, which is a type of sarcopenia. For example, WO2007 / 088123 discloses a muscular atrophy treatment agent containing a nitroxy derivative as an active ingredient, and WO2006 / 081997 discloses a muscular atrophy treatment agent containing atralic acid or its derivative as an active ingredient. However, these treatment agents containing compounds as active ingredients act not only on skeletal muscles affected by muscular atrophy, but also on visceral muscles or cardiac muscles that are not related to muscular atrophy, and therefore may induce various side effects, large or small, and are not used for practical treatment.On the other hand, hormone preparations have significantly fewer side effects than compound preparations and are gentle on the body due to their characteristics, so the development of drugs for treating muscle atrophy or sarcopenia using hormone preparations is accelerating.

[0006] Muscle atrophy can also occur due to factors such as damage to muscle tissue in the absence of mechanical stimulation such as reduced muscle use, muscle destruction due to direct injury or physical factors, impaired muscle cell recovery due to aging, and impaired muscle use due to damage to the nerves that regulate muscle function (Booth FW., J Appl Physiol Respir Environ Exerc Physiol., 1982). In general, disuse atrophy occurs when the muscles in the affected area and surrounding areas are not used for a long period of time due to an injury or accident, leading to a loss of muscle strength and gradual atrophy. It can also occur in the form of myasthenia gravis, which is caused by diseases of the muscles themselves; dystrophy (progressive muscular dystrophy, myotonic muscular dystrophy, Duchenne type, Becker type, limb-girdle type, facioscapulohumeral type); spinal muscular atrophy (Werdnig-Hoffmann type, Kugelberg disease, Welander disease), amyotrophic lateral sclerosis (ALS): Lou Gehrig's disease, and spinal-bulbar muscular atrophy (Kennedy disease), which is caused by inflammation in the muscles themselves and damage to the nerves that control the muscles.

[0007] Such muscle loss is the most common symptom in anti-cancer patients, and occurs when muscles are exposed to an inflammatory state due to the toxicity of the cancer itself or the anti-cancer drug. Muscle loss limits patients' activities and anti-cancer treatment, significantly reducing their quality of life and even causing death, so research into controlling this loss is increasing.

[0008] Meanwhile, macrophages are polarized into anti-inflammatory M2 macrophages and pro-inflammatory M1 macrophages. Of these, M1 macrophages produce pro-inflammatory cytokines such as IL-1, TNF-α, and IL-6, which induce muscle wasting and increase protein degradation through myofiber lysis, while M2 macrophages secrete various anti-inflammatory cytokines such as TGF-β and IL-10 to induce muscle recovery and regeneration and promote muscle fiber synthesis. The balance between M1 and M2 macrophage populations is important for muscle recovery, as macrophage-derived IGF-1 shifts the polarity to M2a and M2c phenotypes, which are macrophage populations that express high levels of matrix metalloproteinase-8 (MMP-8), CD163, and CD206 and are involved in extracellular matrix remodeling and muscle healing, promoting muscle recovery and protecting against muscular dystrophy. [Prior art documents] [Patent documents]

[0009] [Patent Document 1] WO 2007 / 088123-2 [Patent Document 2] WO 2007 / 081997-2 Summary of the Invention [Problem to be solved by the invention]

[0010] An object of the present invention is to provide a pharmaceutical composition for preventing or treating muscle diseases.

[0011] Another object of the present invention is to provide a pharmaceutical composition for preventing or treating side effects of anticancer drugs.

[0012] Another object of the present invention is to provide a food composition for preventing or improving muscle diseases.

[0013] Another object of the present invention is to provide a composition for promoting differentiation of anti-inflammatory macrophages.

[0014] It is also an object of the present invention to provide a method for treating muscle disorders.

[0015] It is also an object of the present invention to provide an application for using the pharmaceutical composition for the prevention and treatment of muscle diseases.

[0016] Another object of the present invention is to provide a method for treating diseases caused by side effects of anticancer drugs.

[0017] Another object of the present invention is to provide a use for producing a pharmaceutical composition for preventing and treating side effects of anti-cancer drugs. [Means for solving the problem]

[0018] To achieve the above object, the present invention provides a pharmaceutical composition for preventing or treating muscular diseases, comprising an extract of Magnolia officinalis as an active ingredient.

[0019] The present invention also provides a pharmaceutical composition for preventing or treating side effects of anti-cancer drugs, comprising an extract of Magnolia officinalis as an active ingredient.

[0020] The present invention also provides a food composition for preventing or improving muscle diseases, which contains Magnolia officinalis extract as an active ingredient.

[0021] The present invention also provides a composition for promoting differentiation of anti-inflammatory macrophages, comprising an extract of Magnolia officinalis as an active ingredient.

[0022] The present invention also provides a method for treating a muscular disease, comprising the step of administering a pharmacopoeias magnolia extract in ...

[0023] The present invention also provides a use of the Magnolia officinalis extract for the manufacture of a pharmaceutical composition for the prevention and treatment of muscle diseases.

[0024] The present invention also provides a method for treating anti-cancer drug side effects, comprising the step of administering a pharmacopoeias magnolia extract in a pharmacopoeias effective amount to an individual having anti-cancer drug side effects.

[0025] In addition, the present invention provides a use of the Magnolia officinalis extract for the manufacture of a pharmaceutical composition for preventing and treating side effects of anti-cancer drugs. Effect of the Invention

[0026] According to the present invention, the Magnolia extract contains various ingredients in addition to magnolol and honokiol, and does not interfere with anti-tumor functions, while suppressing weight loss, muscle loss, grip strength loss and muscle fiber damage caused by cisplatin, and promoting the restoration of damaged muscle fibers through macrophage polarization from M1 to M2, so it can be usefully used in the prevention and treatment of muscle diseases, especially muscle diseases caused by anti-cancer drugs. [Brief description of the drawings]

[0027] [Figure 1] FIG. 1 shows HPLC analysis of the components of Magnolia officinalis extract. [Diagram 2] Figure showing weight loss and food intake changes following administration of Magnolia extract in a cisplatin-induced sarcopenic mouse model: A: weight change; B: weight change compared to the control group; C: colon length; D: average daily food intake; Control (PBS): PBS-administered control group; CIS: cisplatin-administered group; CIS+MAG: cisplatin and magnolol-administered group; and CIS+MC: cisplatin and Magnolia extract-administered group (50, 100 and 200 mg / kg). [Diagram 3]Figure 1 shows the muscle wasting suppression effect of Magnolia extract administration in a cisplatin-induced sarcopenic mouse model: A: limb muscle weight; B: representative limb image (scale bar 0.5 cm); C: TA muscle weight; D: grip strength measured by behavioral skill test in forelimbs and all muscles; E: H&E staining of TA muscle (scale bar 200 μm); F: CSA (cross-sectional area) analysis; Control (PBS): PBS-treated control group; CIS+PBS: cisplatin and PBS-treated group; CIS+MAG: cisplatin and magnolol-treated group; and CIS+MC: cisplatin and Magnolia extract-treated group (50, 100 and 200 mg / kg). [Figure 4] This figure shows that Magnolia officinalis extract regulates M1 and M2 macrophage polarization in skeletal muscle of a mouse model of sarcopenia induced by cisplatin, as confirmed by mRNA expression of M1-specific and M2-specific genes: A and B: M1 macrophage-specific markers; C-F: M2 macrophage-specific markers; Control (PBS): PBS-treated group; CIS+PBS: cisplatin and PBS-treated group; CIS+MAG: cisplatin and magnolol-treated group; and CIS+MC: cisplatin and Magnolia officinalis extract-treated group (50, 100 and 200 mg / kg). [Diagram 5]Figure 1 shows the effect of Magnolia officinalis extract on increasing macrophage number and IGF-1 expression in TA muscle of a mouse model of cisplatin-induced sarcopenia: A: IHC images showing CD68 (macrophage, green), IGF-1 (red), DAPI (nuclei, blue), and a merged image of cells expressing both CD68 and IGF-1 (white arrow) in TA muscle (Magnification: 40×, scale bar 50 μm); B: Number of IGF-I, CD68, and DAPI positive cells; C: IGF-1 mRNA expression level; D: IGF-1 protein expression level; Control (PBS): PBS-treated group; CIS+PBS: cisplatin and PBS-treated group; CIS+MAG: cisplatin and magnolol-treated group; and CIS+MC: cisplatin and Magnolia officinalis extract-treated group (50, 100, and 200 mg / kg). [Figure 6] Figures confirming the effect of Kobo extract in increasing the percentage of M2a and M2c macrophages in a cisplatin-induced sarcopenic mouse model: A and B: FACS dot plots of CD206 and CD163 in CD11b+F4 / 80+ and CD45+ cells (CD206-CD163-: M1 macrophages; CD206+CD163-: M2a macrophages; and CD206+CD163+: M2c macrophages); C-E: M1 macrophages, M2a macrophages and M2c macrophages graphs (shown as % of CD11b+F4 / 80+ cells); F: CD4+ / CD8 ratio of CD45+ cells in spleen cells; Control (PBS): PBS-treated control group; CIS+PBS: cisplatin and PBS-treated group; CIS+MAG: cisplatin and magnolol-treated group; and CIS+MC: cisplatin and Magnolia extract-treated group (50, 100 and 200mg / kg). [Figure 7] This figure confirms the effect of Magnolia officinalis extract on the antitumor activity of cisplatin: A: tumor size; B: tumor image; C: tumor weight; Control: control group; CIS: cisplatin-administered group; and CIS+MC: cisplatin and Magnolia officinalis extract-administered group. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0028] Hereinafter, the present invention will be described in detail with reference to the accompanying drawings. However, the following embodiments are presented as examples of the present invention, and if it is determined that a detailed description of a well-known technology or configuration may obscure the gist of the present invention, the detailed description may be omitted and the present invention is not limited thereby. The present invention is subject to various modifications and applications within the scope of the claims and the scope of equivalents analyzed therefrom.

[0029] In addition, the terminology used in this specification is used to appropriately express the preferred embodiment of the present invention, and may vary depending on the intention of the user or operator, or the practice of the field to which the present invention belongs. Therefore, the definition of the term should be based on the overall content of this specification. Throughout the specification, when a part "includes" a certain element, this does not mean that other elements are excluded, but that other elements may be further included, unless otherwise specified.

[0030] Throughout this specification, "%" used to indicate the concentration of a particular substance is (w / w)% for solid / solid, (w / v)% for solid / liquid, and (v / v)% for liquid / liquid, unless otherwise stated.

[0031] In one aspect, the present invention relates to a pharmaceutical composition for preventing or treating a muscular disease, comprising an extract of Magnolia officinalis as an active ingredient.

[0032] In one embodiment, the muscle disease may be a muscle disease caused by muscle loss, muscle tissue damage, muscle wasting or muscle degeneration, or may be a muscle disease caused by cancer.

[0033] In one embodiment, the muscle disease may be any one or more selected from the group consisting of muscular atrophy, myopathy, muscle degeneration, myasthenia, muscular injury, dystrophinopathy, myopathy, muscular dystrophy, cachexia, and sacopenia.

[0034] In one embodiment, the extract may be extracted with one or more solvents selected from the group consisting of water, organic solvents, subcritical fluids, and supercritical fluids, and the organic solvent may be any one selected from the group consisting of lower alcohols having 1 to 4 carbon atoms, hexane (n-hexane), ether, glycerol, propylene glycol, butylene glycol, ethyl acetate, methyl acetate, dichloromethane, chloroform, ethyl acetate, acetone, methylene chloride, cyclohexane, petroleum ether, benzene, and mixtures thereof; In one embodiment, the Magnolia officinalis extract may be an ethanol extract, more preferably a 70% ethanol extract.

[0035] In one embodiment, magnolol and honokiol may be contained in a ratio of 2:1 to 4:1 (w / w).

[0036] In one embodiment of the present invention, the Magnolia extract contains various components in addition to magnolol and honokiol (see Figure 1), and has a significant therapeutic effect on muscle diseases, particularly muscle diseases induced by administration of anti-cancer drugs, at concentrations exceeding 50 mg / kg, compared to administration of magnolol alone.

[0037] In one embodiment, the composition may contain Magnolia officinalis extract at a concentration of 60-500 mg / kg.

[0038] In one implementation, the composition can inhibit weight loss, muscle mass loss, grip strength loss, or muscle fiber damage.

[0039] The term "extract" used in the present invention means an active ingredient separated from a natural product, i.e., a substance exhibiting a desired activity. The extract may be obtained by an extraction process using water, an organic solvent, or a mixture thereof, and includes all forms of the extract, its dried powder, or formulations using these. The extract also includes fractions of the extract obtained by the extraction process. The method of extracting the extract is not particularly limited, and may be, for example, a method such as stirring extraction, shaking extraction, hot water extraction, cold immersion extraction, reflux cooling extraction, or ultrasonic extraction. The extraction solvent may be a polar solvent such as water or a C1-C4 lower alcohol, or a non-polar solvent such as hexane, chloroform, dichloromethane, or ethyl acetate, or a mixture of two or more of these.

[0040] The composition of the present invention can be prepared as a pharmaceutical composition for preventing or treating muscle diseases by containing not only Magnolia officinalis extract but also other active ingredients having the same or similar functions, or by containing other active ingredients having different functions from the above-mentioned ingredients.

[0041] In one aspect, the present invention relates to a pharmaceutical composition for preventing or treating side effects of anti-cancer drugs, comprising an extract of Magnolia officinalis as an active ingredient.

[0042] In one embodiment, the anticancer drug may be cyclophosphamide, methotrexate, 5-fluorouracil, doxorubicin, mustine, vincristine, procarbazine, prednisolone, bleomycin, vinblastine, dacarbazine, etoposide, cisplatin, epirubicin, cisplatin, capecitabine, or oxaliplatin, and is more preferably cisplatin.

[0043] In one embodiment, the anti-cancer drug side effect disease may be a muscle disease caused by an anti-cancer drug side effect, and is more preferably any one or more selected from the group consisting of muscle atrophy, muscle degeneration, muscle damage, muscular dystrophy, cachexia, and sarcopenia caused by an anti-cancer drug side effect, and is even more preferably cachexia or sarcopenia caused by an anti-cancer drug side effect.

[0044] In one embodiment, the composition can inhibit weight loss, muscle mass loss, grip strength loss, or muscle fiber damage caused by anti-cancer drugs.

[0045] In one implementation, the composition can be administered separately, simultaneously or sequentially with the anti-cancer drug.

[0046] In one embodiment, the pharmaceutical composition for preventing or treating anti-cancer drug side effects can be used as an anti-cancer adjuvant.

[0047] In one embodiment, the composition can promote the restoration of muscle fibers damaged by anti-cancer drugs through macrophage polarization from M1 to M2.

[0048] In the present invention, the term "prevention" means any action of suppressing or delaying the occurrence, spread and recurrence of muscle disease or anti-cancer drug side effect disease by administering the pharmaceutical composition of the present invention, and "treatment" means any action of improving or favorably changing the symptoms of muscle disease or anti-cancer drug side effect disease by administering the composition of the present invention. Those with ordinary skill in the art to which the present invention pertains should be able to refer to materials presented by the Korean Medical Association, etc., to know the exact criteria for diseases for which the composition of the present application is effective, and to judge the degree of improvement, enhancement and treatment.

[0049] The term "therapeutically effective amount" used in combination with the active ingredient in the present invention means an amount effective for preventing or treating muscle disease or anticancer drug side effect disease, and the therapeutically effective amount of the composition of the present invention may vary depending on several factors, such as the administration method, the target site, and the condition of the patient. Therefore, the dosage when used in the human body must be determined as an appropriate amount by considering both safety and efficacy. It is also possible to estimate the amount used in humans from the effective amount determined through animal experiments. These matters to be considered when determining the effective amount are described, for example, in Hardman and Limbird, eds., Goodman and Gilman's The Pharmacological Basis of Therapeutics, 10th ed.(2001), Pergamon Press; and EW Martin ed., Remington's Pharmaceutical Sciences, 18th ed.(1990), Mack Publishing Co.

[0050] The pharmaceutical composition of the present invention is administered in a pharmacologic effective amount. The term "pharmacologic effective amount" used in the present invention means an amount sufficient to treat a disease with a reasonable benefit / risk ratio applicable to medical treatment and not to cause side effects, and the effective dose level can be determined based on factors including the patient's health condition, the type of muscle disease or anticancer drug side effect disease, the cause of onset and severity of the muscle disease or anticancer drug side effect disease, the activity of the drug, the sensitivity to the drug, the administration method, administration time, administration route and excretion rate, treatment period, drugs used in combination or simultaneously, and other factors well known in the medical field. The composition of the present invention can be administered as an individual therapeutic agent or in combination with other therapeutic agents, and can be administered sequentially or simultaneously with conventional therapeutic agents, and can be administered singly or multiple times. Taking all of the above factors into consideration, it is important to administer an amount that can obtain the maximum effect with the minimum amount without side effects, which can be easily determined by those skilled in the art.

[0051] The pharmaceutical composition of the present invention may contain carriers, diluents, excipients, or combinations of two or more thereof that are commonly used in biological preparations. The term "pharmaceutical acceptable" as used herein means that the composition exhibits the property of being non-toxic to cells or humans exposed to the composition. The carrier is not particularly limited as long as it is suitable for in vivo delivery of the composition, and may be, for example, a compound listed in the Merck Index, 13th ed., Merck & Co. Inc., saline, sterile water, Ringer's solution, buffered saline, dextrose solution, maltodextrin solution, glycerol, ethanol, or a mixture of one or more of these components, and other common additives such as antioxidants, buffers, bacteriostatic agents, etc. may be added as necessary. In addition, the composition may be formulated into an injectable dosage form such as an aqueous solution, suspension, emulsion, etc., pills, capsules, granules, or tablets by additionally adding a diluent, dispersant, surfactant, binder, and lubricant. Furthermore, formulations can be suitably prepared according to each disease or ingredient using a method appropriate to the art or a method disclosed in Remington's Pharmaceutical Sciences (Mack Publishing Company, Easton PA, 18th, 1990).

[0052] In one embodiment, the pharmaceutical composition may be in one or more dosage forms selected from the group including oral dosage forms, topical preparations, suppositories, sterile injection solutions and sprays, with oral or injection dosage forms being more preferred.

[0053] The term "administration" as used in the present invention means providing a given substance to an individual or patient in any suitable manner, and can be administered parenterally (e.g., intravenously, subcutaneously, intraperitoneally, or locally in the form of an injection) or orally depending on the intended method, and the dosage range varies depending on the patient's weight, age, sex, health condition, food, administration time, administration method, excretion rate, and severity of disease. Liquid preparations for oral administration of the composition of the present invention include suspensions, oral liquids, emulsions, syrups, etc., which may contain various excipients such as wetting agents, sweeteners, flavoring agents, preservatives, etc. in addition to water and liquid paraffin, which are commonly used simple diluents. Preparations for parenteral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized preparations, suppositories, etc. The pharmaceutical composition of the present invention can be administered by any device that can transfer the active substance to target cells. Preferred administration methods and preparations are intravenous injections, subcutaneous injections, intradermal injections, intramuscular injections, drip injections, etc. Injectables can be prepared using aqueous solvents such as physiological saline and Ringer's solution, and non-aqueous solvents such as vegetable oils, higher fatty acid esters (e.g., ethyl oleate, etc.), and alcohols (e.g., ethanol, benzyl alcohol, propylene glycol, glycerin, etc.), and can contain pharmaceutical carriers such as stabilizers to prevent deterioration (e.g., ascorbic acid, sodium bisulfite, sodium pyrosulfite, BHA, tocopherol, EDTA, etc.), emulsifiers, buffers for pH adjustment, and preservatives to prevent microbial growth (e.g., phenylmercuric nitrate, thimerosal, benzalkonium chloride, phenol, cresol, benzyl alcohol, etc.).

[0054] The term "individual" used in the present invention means any animal, including humans, monkeys, cows, horses, goats, pigs, chickens, turkeys, quails, cats, dogs, mice, rats, rabbits, or guinea pigs, who have or may develop the muscle disease or anticancer drug side effect disease, and the pharmaceutical composition of the present invention can be administered to an individual to effectively prevent or treat the disease. The pharmaceutical composition of the present invention can be administered in parallel with an existing therapeutic agent.

[0055] The pharmaceutical composition of the present invention may further include a pharma- ceutically acceptable additive, and examples of the pharma-ceutically acceptable additive include starch, gelatinized starch, microcrystalline cellulose, lactose, povidone, colloidal silicon dioxide, calcium hydrogen phosphate, lactose, mannitol, candy, gum arabic, pregelatinized starch, corn starch, powdered cellulose, hydroxypropyl cellulose, opadry, sodium starch glycolate, carnauba wax, synthetic aluminum silicate, stearic acid, magnesium stearate, aluminum stearate, calcium stearate, sucrose, dextrose, sorbitol, and talc. The pharma-ceutically acceptable additive according to the present invention is preferably included in an amount of 0.1 to 90 parts by weight based on the composition, but is not limited thereto.

[0056] In one aspect, the present invention relates to a food composition for preventing or improving a muscle disease, comprising Magnolia officinalis extract as an active ingredient.

[0057] In one embodiment, the muscle disease may be a muscle disease caused by cancer or a muscle disease caused by anti-cancer drug treatment.

[0058] In one embodiment, the muscle disease may be any one or more selected from the group consisting of muscular atrophy, myopathy, muscle degeneration, myasthenia, muscular injury, dystrophinopathy, myopathy, muscular dystrophy, cachexia, and sacopenia.

[0059] When the composition of the present invention is used as a food composition, the composition can be added as it is or can be used together with other foods or food ingredients, and can be used appropriately by a conventional method. The composition can contain a food-scientifically acceptable food supplement additive in addition to the active ingredient, and the amount of the active ingredient to be mixed can be appropriately determined depending on the purpose of use (prevention, health or therapeutic treatment).

[0060] The term "food supplement additive" used in the present invention means a component that can be added to food as a supplement, and can be appropriately selected and used by those skilled in the art as an additive to produce health functional food of various dosage forms. Examples of food supplement additives include various nutrients, vitamins, minerals (electrolytes), flavors such as synthetic flavors and natural flavors, colorants and fillers, pectinic acid and its salts, alginic acid and its salts, organic acids, protective colloid thickeners, pH regulators, stabilizers, preservatives, glycerin, alcohol, carbonation agents used in carbonated drinks, etc., but the above examples do not limit the types of food supplement additives of the present invention.

[0061] The food composition of the present invention may include a functional health food. The term "functional health food" used in the present invention refers to a food manufactured and processed in the form of tablets, capsules, powders, granules, liquids, pills, etc., using raw materials or ingredients having useful functions for the human body. Here, "functionality" means that it regulates nutrients for the structure and function of the human body, or obtains useful effects for health applications such as physiological action. The functional health food of the present invention can be manufactured by a method commonly used in the ordinary technical field, and during the manufacturing process, it can be manufactured by adding raw materials and ingredients that are commonly added in the ordinary technical field. In addition, the dosage form of the functional health food can also be manufactured without any restrictions as long as it is a dosage form recognized as a functional health food. The food composition of the present invention can be manufactured in various dosage forms, and unlike general medicines, it has the advantage of being made of food as a raw material and having no side effects that may occur when taking medicines for a long period of time, and is highly portable. The functional health food of the present invention can be taken as an adjuvant to enhance the effect of anticancer drugs.

[0062] In addition, there is no limit to the type of health food in which the composition of the present invention can be used. In addition, the composition containing the Magnolia officinalis extract of the present invention as an active ingredient can be prepared by mixing other appropriate auxiliary ingredients and known additives that can be included in health functional foods as selected by those skilled in the art. Examples of foods that can be added include meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, gums, dairy products including ice cream, various soups, beverages, tea, energy drinks, alcoholic beverages, and vitamin complexes, and can be prepared by adding the extract of the present invention to soups, teas, jellies, and juices made as the main ingredient.

[0063] In one aspect, the present invention relates to a composition for promoting anti-inflammatory macrophage differentiation, comprising an extract of Magnolia officinalis as an active ingredient.

[0064] In one embodiment, the composition can induce M2 macrophage polarization and increase expression of IGF-1 (IGF-1-like growth factor-1) expression.

[0065] In one embodiment, the composition comprises a polypeptide that binds to M1 macrophages (CD206 + CD163 - ) population and reduced M2a macrophages (CD206 + CD163 - ) population and M2c macrophages (CD206 + CD163 + ) population can be increased.

[0066] In one aspect, the present invention relates to a method for treating a muscular disorder, comprising administering a pharmacopoeias magnolia extract in ...

[0067] In one aspect, the present invention relates to the use of an extract of Magnolia officinalis for the manufacture of a pharmaceutical composition for the prevention and treatment of muscle diseases.

[0068] In one aspect, the present invention relates to a method for treating anti-cancer drug side effect disorders, comprising the step of administering a pharmacopoeias magnolia extract in a pharmacopoeias effective amount to an individual having anti-cancer drug side effect disorders.

[0069] In one embodiment, the anticancer drug side effect disease may be a muscle disease, and more preferably is any one or more selected from the group consisting of muscle atrophy, muscle degeneration, muscle damage, muscular dystrophy, cachexia and sarcopenia.

[0070] In one aspect, the present invention relates to use of an extract of Magnolia officinalis for the manufacture of a pharmaceutical composition for the prevention and treatment of side effects of anti-cancer drugs.

[0071] The present invention will be described in more detail with reference to the following examples, which are provided for illustrative purposes only and are not intended to limit the scope of the present invention.

[0072] Example 1. Preparation of Magnolia officinalis extract

[0073] 1000mL of ethanol:water (7:3, v / v) mixed solvent (70% ethanol) was mixed with 200g of Magnolia officinalis (Rehder et Wilson) (Sunchen, Korea), extracted twice in a reflux system at 70℃, filtered, concentrated under reduced pressure at 40~45℃, and freeze-dried for 3 days to produce Magnolia officinalis extract (MC) with a yield of 10.91% (21.82g).

[0074] Example 2. Analysis of Magnolia Extract

[0075] The components of the ethanol extract of Magnolia officinalis prepared in Example 1 were analyzed using HPLC, and magnolol and honokiol present in the ethanol extract of Magnolia officinalis were analyzed at Dong-eul Herbal Medicine Analysis Center (Busan, Korea). Specifically, magnolol and honokiol were analyzed by ODS (octadecyl silica)-chromatography using a DHAC-12-01 (Agilent, Santa Clara, CA, USA) equipped with a UV-vis spectrophotometer (289 nm). Magnolol and honokiol were analyzed by isocratic elution using acetonitrile:distilled water:acetic acid (70:30:1, v / v / v) as the mobile phase components at 20°C using an OSAKASODA C18 column (5um particle size, 4.6×250mm; Osaka, Japan), with injection volume and flow rate of 10μL and 0.3mL / min, respectively. Magnolol and honokiol standards were detected at 289nm for quantitative HPLC analysis. In addition, the freeze-dried powder (0.1g) of the Magnolia ethanol extract prepared in Example 1 was dissolved in methanol (100mL), sonicated for 20 minutes, filtered, and then used for ODS-HPLC analysis. Magnolol and honokiol standards were also dissolved in 100% methanol at a concentration of 100mg / L, respectively, and HPLC analysis was performed.

[0076] As a result, it was confirmed that the magnolol and honokiol contents in the magnolia ethanol extract (MC) were 2.1% and 0.8%, respectively (Figure 1).

[0077] Example 3. Inhibitory effect of Magnolia extract on cisplatin-mediated weight loss

[0078] In order to confirm the effect of Magnolia Magnolia extract on muscle atrophy and rapid weight loss caused by cisplatin, a common chemotherapy anticancer treatment, mice were administered cisplatin to induce weight loss, and the effect was confirmed by administering Magnolia Magnolia ethanol extract (MC) of Example 1. Specifically, mice (C57BL / 6, 7-8 weeks old) (Jackson Laboratory, Gyeonggi Raon Bio) were randomly housed in cages in a regulated environment with a 12-hour light-dark cycle and free access to food and water, and then cisplatin (Sigma-Aldrich, P4394; St. Louis, MO, USA) (2.5 mg / kg) dissolved in saline at 1 mg / mL was intraperitoneally injected once a day on days 1-5 and 26-30 (sarcopenic mouse model), and MC (50, 100 and 200 mg / kg) was orally administered every 3 days (total of 12 times). The control and negative control groups (CIS+PBS) were orally administered PBS every 3 days, and the positive control group was intraperitoneally injected with magnolol (purity 95%) (Sigma (m3445; Sigma-Aldrich, St. Louis, MO, USA) (10 mg / kg) dissolved in DMSO at 10 mM every 3 days (total 12 times). All experimental procedures were carried out by the Kyung Hee Institutional Animal Care and Use Center (CACHC). The study was conducted with the approval of the FDA (KHUASP(SE)-20-529). As a result, cisplatin administration induced rapid weight loss in mice from the 15th day, which was significantly alleviated by MC or magnolol administration, showing rapid weight recovery (Figure 2). This significant difference between the CIS(cisplatin)+PBS and CIS+MC groups continued for more than 6 days, and the mice's weight decreased by more than 30%, and the experiment was terminated on the 42nd day (Figure 2A). In particular, the group administered MC at 100 or 200 mg / kg showed a more significant suppression of cisplatin-induced weight loss than the magnolol-administered group. In addition, the results of checking the weight change on the last and first days showed that the mice administered cisplatin showed a significant weight loss, while the MC or magnolol-administered groups significantly reduced the weight loss induced by cisplatin (Figure 2B).To determine whether this weight loss was related to impaired digestion and appetite, colon length and average daily food intake were measured. Results showed that there were no differences in colon length and average daily food intake between the groups (Figures 2C and D), indicating that the cisplatin-induced weight loss was not related to abnormal digestive or appetite changes.

[0079] Example 4. Cisplatin-induced skeletal muscle wasting alleviation effect of Magnolia extract

[0080] 4-1. Confirmation of muscle mass loss suppression effect

[0081] To confirm the effect of Magnolia officinalis extract on cisplatin-induced skeletal muscle wasting, the limb weight and tibialis anterior (TA) muscle mass of each group were measured in experiment 42. The results showed that both the MC and magnolol groups significantly alleviated the limb weight and TA muscle mass losses induced by cisplatin, and this effect was significantly higher in the MC group than in the magnolol group (Figure 3A-C).

[0082] 4-2. Confirmation of the effect of suppressing the decline in grip strength

[0083] To confirm the effect of Magnolia extract on the reduction in grip strength caused by cisplatin, muscle strength of all limbs and forelimbs was measured using a digital force gauge (DS2-5N; IMADA Inc., Northbrook, IL, USA). Muscle strength was measured by pulling the tail of the mouse downwards five times and recording the peak tension when the mouse placed the limb down. The muscle strength was measured three times at 10-min intervals, and the average of the five values ​​was used for calculation. The results showed that the pulling force power was significantly increased in the MC (100 and 200 mg / kg) and magnolol groups compared to the cisplatin group, and this effect was significantly higher in the MC 100 mg / kg or 200 mg / kg groups compared to the magnolol group (Figure 3D).

[0084] 4-3. Confirmation of muscle fiber damage suppression effect

[0085] To confirm the effect of Magnolia officinalis extract on muscle fiber damage caused by cisplatin, the muscle fiber morphology of skeletal muscle was examined through H&E (hematoxylin and eosin) staining. Specifically, TA muscle tissue slices were fixed in 4% formalin for 24 hours, embedded in paraffin, and sectioned at a thickness of 4 μm. The sections were treated as described by Lee et al., H&E stained to evaluate the pathological changes of the tissue, and imaged with an Eclipse Ci-L microscope (Nikon, Tokyo, Japan). The results showed that muscle fiber deformation (muscle fiber damage) was prevented in both the 100 and 200 mg / kg MC groups, unlike the 50 mg / kg MC group (Figure 3E), confirming that Magnolia officinalis extract prevents or inhibits muscle loss and damage in mice with cisplatin-induced sarcopenia.

[0086] Example 5. Polarization and activation effect of M1 and M2 macrophages in skeletal muscle

[0087] Since the restoration of damaged muscle fibers requires the activation of M2 macrophages, we confirmed by qPCR (real-time quantitative PCR) whether the administration of Magnolia extract changes the expression of macrophage-specific marker genes, including Arg-1, NOS2, TNF-α, MRC1, TGF-β, and CD163, in skeletal muscle. Specifically, total RNA was extracted from TA muscle using easy-BLUE TM Total RNA Extraction Kit (17061; iNtRON, Biotechnology, Jungwon, Korea), and cDNA was synthesized using Maxime RT-PCR PreMix Kit (25131; iNtRON Biotechnology, Jungwon, Korea). Real-time qPCR was then performed using the primer set (5'→3') for each gene below and SensiFAST SYBR No-ROX Kit (BIO-98020; Medison bioline, Rome, Italy), and the results were compared with standard 2 δδCtmethod and normalized with GAPDH (glyceraldehyde-3-phosphate dehydrogenase): Nos2 (Forward: CAC CTT GGA GTT CAC CCA GT, Reverse: ACC ACT CGT ACT TGG GAT GC), Igf-1 (Forward: CTA CCA AAA TGA CCG CAC CT, Reverse: CAC GAA CTG AAG AGC ATC CA), Tgfb1(Forward:CAA GGA AGG TTG GCA TTT GT, Reverse:AGG TAA CGC CAG GAA TTG CA), Tnfa(Forward:GCT GAG CTC AAA CCC TGG TA, Reverse:CCG GAC TCC GCA AAGTCT AA), Mrc1(Forward:CAA GGA AGG TTG GCA TTT GT, Reverse:CCT TTC AGT CCT TTG CAA GC), Arg-1(Forward:GGC TGG TCT GCT TGA GAA AC, Reverse:CTT TTC CCA CAG ACC TTG GA), Cd163(Forward:TGG TGT GCA GGG AAT TAC AA, Reverse:ATC CCT GCT GTG GGT ACA AG) and Gapdh(Forward:ACC CAG AAG ACT GTG GAT GG, Reverse:CAC ATT GGG GGT AGG AAC AC).

[0088] The results showed that the expression of M1 macrophage-specific marker genes (TNF-α and iNOS) was increased in the cisplatin-treated group, but was significantly decreased by MC treatment (Figures 4A and B). At the same time, the expression levels of M2 macrophage-specific marker genes (CD206, Arg-1, TGF-β, and CD163) were significantly increased in the MC-treated group compared to the cisplatin-treated group (Figures 4C-F). This confirmed that Magnolia extract regulates M1 and M2 macrophage polarization in the skeletal muscle microenvironment.

[0089] Example 6. Effect of increasing macrophage count and IGF-1 expression in skeletal muscle

[0090] 6-1. Confirmation of macrophage count in skeletal muscle

[0091] We examined whether administration of Magnolia officinalis extract affects IGF-1 (IGF-1-like growth factor-1) expression and the number of macrophages in skeletal muscle, which shifts the polarization of macrophages from M2a to M2c (macrophages involved in extracellular matrix remodeling and muscle healing). Specifically, the TA muscle tissue sections of Example 4-3 were incubated overnight with primary antibodies, Rett anti-mouse CD68 antibody (1:250; MCA1957GA; Bio-Rad, Contra Costa County, CA, USA) and rabbit anti-mouse IGF-1 antibody (1:500; 40657; Abcam, Cambridge, UK), and then incubated for 30 minutes with secondary antibodies, Alexa Fluor 488-conjugated goat anti-Rett immunoglobulin IgG (A11006; Invitrogen, Carlsbad, CA, USA) and Alexa Fluor 594-conjugated goat anti-rabbit IgG (A32740; Invitrogen, Carlsbad, CA, USA). The sections were then mounted using mounting media containing DAPI (4',6-diamidino-2-phenylindole) (H-1200, Vector, Torrance, CA, USA) and imaged under a confocal microscope (FV10C-PSU; Olympus Corporation, Tokyo, Japan). The numbers of IGF-I (red), CD68 (macrophage) (green), and DAPI (blue) positive cells per image area were counted in a minimum of five random fields.

[0092] The results showed that the number of cells expressing both CD68 and IGF-1 increased in both the MC-administered and magnolol-administered groups (FIGS. 5A and B).

[0093] 6-2. Confirmation of IGF-1 expression in skeletal muscle

[0094] The IGF-1 mRNA expression level in the TA muscle tissue slices of Example 4-3 was confirmed by qPCR, and the IGF-1 protein expression level was confirmed by ELISA. Specifically, qPCR was performed as in Example 5, and ELISA was performed by dissolving the TA muscle tissue slices in PRO-PREP Protein Extraction Solution (17081; iNtRON Biotechnology, Jungwon, Korea) containing a protease inhibitor cocktail, and then homogenizing them using a mechanical homogenizer (Precellys R24; Bertin, Montigny-le-Bretonneux, France). Then, ELISA analysis was performed using an IGF-1 ELISA kit (DY791; R&D Systems, Minneapolis, MN, USA).

[0095] The results showed that both IGF-1 mRNA and protein expression were significantly increased in the MC-administered group compared with the cisplatin-administered group (Fig. 5C and D).

[0096] Through this, it was confirmed that Magnolia officinalis extract increases the expression of macrophage-derived IGF-1 in skeletal muscles, shifting polarization to the M2a and M2c phenotypes, thereby promoting muscle recovery after cisplatin-induced skeletal muscle injury and protecting against muscular dystrophy.

[0097] Example 7. Macrophage subtype changing effect

[0098] To confirm the effect of Magnolia officinalis extract on the phenotypic change of M1 macrophages, spleen-derived macrophages were extracted from each group of sarcopenic mouse models in Example 3, and the macrophage subtypes were confirmed by flow cytometry. Specifically, the spleens of each group of mice were removed and separated into single cells using a 40 μm nylon mesh strainer. Then, red blood cells were lysed with Pharmlyse buffer (555899; BD bioscience, San Jose, CA, USA), and single cells were incubated with 100 mM NaCl and 100 mM NaCl for 1 h at 4 °C for 1 h. The cells were then incubated for 1 h at 4 °C for 2 h at 4 °C for 1 ... Cells were stained with PE antibody (12-1631-82; e-bioscience; Thermo Fisher Scientific, Middlesex County, MA, USA) and anti-mCD206 APC antibody (141707; BioLegend, San Diego, CA, USA) by incubation at 4°C for 1 h. Stained cells were washed twice with FACS buffer and then analyzed with FACS Lyric instruments (BD bioscience, San Jose, CA, USA). Data were analyzed with FlowJo V10 software (Treestar Inc., San Carlos, CA, USA). Unstained cells (negative control group) were used as a gating control group. The phenotype of the cells is as follows (Figure 6A): CD45 + and CD11b + F4 / 80 + Among the cells gated on the , M1 macrophages were CD206 - CD163 -M2a macrophages are CD206 + CD163 - M2c macrophages express CD206 + CD163 + cell.

[0099] As a result, treatment with Magnolia extract increased the expression of CD11b + F4 / 80 + M1 macrophages (CD206 - CD163 - ) population was significantly decreased compared to the cisplatin-treated group (Fig. 6B and C). In addition, the M2a macrophage (CD206 + CD163 - ) population and M2c macrophages (CD206 + CD163 + ) population was significantly increased (Fig. 6D and E). + / CD8 + Measurement of the T cell percentage showed no difference between the groups (Figure 6F).

[0100] Example 8. Confirmation of the effect of Magnolia extract on the antitumor activity of cisplatin

[0101] To confirm whether administration of Magnolia officinalis extract inhibits the antitumor activity of cisplatin, mouse colon cancer cell line CT26 (Korean Cell Line Bank, 80009; Seoul, Korea) was administered at 3 × 10 per mouse. 5 Mice bearing colon cancer were then subcutaneously injected with 100 mg / kg each to create a mouse model, which was then treated with cisplatin or cisplatin plus Magnolia officinalis extract (200 mg / kg) and the size (volume) and weight of the tumors were measured every 3 days. As a result, cisplatin significantly reduced tumor growth, and the group administered Magnolia officinalis extract together with cisplatin showed a similar tumor growth suppression effect to that of cisplatin alone (Figure 7), confirming that Magnolia officinalis extract does not interfere with the anti-tumor function of cisplatin.

[0102] Through this, it was confirmed that Magnolia extract has preventive and therapeutic effects on cancer cachexia, especially cisplatin-induced sarcopenia, and does not interfere with the anti-tumor effects of cisplatin.

Claims

1. A pharmaceutical composition for preventing or treating muscular diseases, comprising an extract of Magnolia officinalis as an active ingredient.

2. The pharmaceutical composition for preventing or treating a muscle disease according to claim 1, wherein the muscle disease is caused by reduced muscle function, muscle tissue damage, muscle wasting or muscle degeneration.

3. The pharmaceutical composition for preventing or treating a muscular disease according to claim 1, which is a muscular disease caused by cancer.

4. 2. The pharmaceutical composition for preventing or treating a muscular disease according to claim 1, wherein the muscular disease is at least one selected from the group consisting of muscular atrophy, myopathy, muscular degeneration, myasthenia, muscular injury, dystrophinopathy, myopathy, muscular dystrophy, cachexia, and saccopenia.

5. The pharmaceutical composition for preventing or treating a muscle disease according to claim 1, wherein the extract is extracted with one or more solvents selected from the group consisting of water, an organic solvent, a subcritical fluid, and a supercritical fluid.

6. 2. The pharmaceutical composition for preventing or treating muscular diseases according to claim 1, wherein the magnolia extract contains magnolol and honokiol in a ratio of 2:1 to 4:1 (w / w).

7. 2. The pharmaceutical composition for preventing or treating muscular diseases according to claim 1, comprising the Magnolia officinalis extract in a concentration of 60 to 500 mg / kg.

8. The pharmaceutical composition for preventing or treating a muscular disease according to claim 1, which suppresses weight loss, muscle mass loss, grip strength loss, or muscle fiber damage.

9. A pharmaceutical composition for preventing or treating side effects of anti-cancer drugs, comprising an extract of Magnolia officinalis as an active ingredient.

10. Anticancer drugs include cyclophosphamide, methotrexate, 5-fluorouracil, doxorubicin, mustine, vincristine, procarbazine, prednisolone, and bleomycin.

10. The pharmaceutical composition for preventing or treating anticancer drug side effect diseases according to claim 9, wherein the active ingredient is cisplatin, vinblastine, dacarbazine, etoposide, cisplatin, epirubicin, cisplatin, capecitabine or oxaliplatin.

11. The pharmaceutical composition for preventing or treating anticancer drug side effects according to claim 9, wherein the anticancer drug side effect disease is any one or more selected from the group consisting of muscle atrophy, muscle degeneration, muscle damage, muscular dystrophy, cachexia and sarcopenia.

12. The pharmaceutical composition for preventing or treating side effects of anticancer drugs according to claim 9, which suppresses weight loss, muscle mass loss, grip strength loss, or muscle fiber damage caused by anticancer drugs.

13. The pharmaceutical composition for preventing or treating side effects of anti-cancer drugs according to claim 9 , which is administered separately, simultaneously or sequentially with an anti-cancer drug.

14. A food composition for preventing or improving muscle diseases, comprising Magnolia officinalis extract as an active ingredient.

15. A composition for promoting differentiation of anti-inflammatory macrophages, comprising Magnolia officinalis extract as an active ingredient.

16. The composition for promoting anti-inflammatory macrophage differentiation according to claim 15, which induces M2 macrophage polarization.

17. A method for treating a muscular disease, comprising administering a pharmacopoeias extract in a pharmacopoeias medica ...

18. Use of Magnolia officinalis extract for the manufacture of a pharmaceutical composition for the prevention and treatment of muscle diseases.

19. A method for treating anti-cancer drug side effects, comprising the step of administering a pharmacopoeias extract in a pharmacopoeias to an individual suffering from anti-cancer drug side effects.

20. The method for treating anticancer drug side effect diseases according to claim 19, wherein the anticancer drug side effect disease is any one or more selected from the group consisting of muscle atrophy, muscle degeneration, muscle damage, muscular dystrophy, cachexia and sarcopenia.

21. Use of Magnolia officinalis extract for the manufacture of a pharmaceutical composition for the prevention and treatment of side effects of anti-cancer drugs.

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