WO2020017921A1 - Pharmaceutical composition, for preventing or treating sarcopenia, containing salicornia europaea extract - Google Patents

Abstract

According to an embodiment of the present invention, provided are a pharmaceutical composition, for preventing or treating sarcopenia, containing a Salicornia europaea extract as an active ingredient, and a food composition, for preventing or alleviating sarcopenia, containing a Salicornia europaea extract as an active ingredient.

Description

Pharmaceutical composition for the prevention or treatment of myopathy containing hamwort extract

The present invention relates to a pharmaceutical composition for the prevention or treatment of myopathy, which contains a seaweed extract as an active ingredient.

The concept of sarcopenia began with the introduction of the term 'sarcopenia' by Irwin Rosenberg in 1989, where the Greek word "sarco" means muscle and "penia" means reduced. Muscular dystrophy refers to the decrease in muscle strength associated with aging with a decrease in muscle mass. "Muscle" here means skeletal muscle and has nothing to do with smooth muscle. In other words, myotropia mainly refers to a loss of skeletal muscle mass distributed in the extremities, and muscle loss due to acute diseases such as cachexia and flu, which are significant muscle loss states in the late stages of malignant tumors. wasting, or primary muscle disease.

Recently, the prevalence of osteoporosis and muscular dystrophy is increasing rapidly as the age group of 65 and older increases rapidly. It is estimated that a gradual decrease in muscle mass occurs after the 40s and decreases by 8% every 10 years until the 70s. After that, it is known that a more rapid decrease may occur and may occur up to 15% every 10 years. have. Many follow-up studies have shown that the physiological changes that occur in older people vary and, in general, decrease in muscle mass and bone density with age.

On the other hand, there are three major methods of treatment of muscular dystrophy. The first is exercise. Exercise has been reported to increase skeletal muscle protein synthesis in the short term and increase muscle strength or mobility in older adults. However, it is inadequate for long-term treatment. Second, testosterone or anabolic steroid can be used as a drug treatment, but this induces masculinity in females, and in men, side effects such as prostate symptoms. Other approved regimens include dehydroepiandrosterone (DHEA) and growth hormone, which have been reported to be therapeutic in areas containing SARMs (Selective Androgen Receptor Modulators). In addition, although diet is known as a treatment, nutritional assessments indicate that malnutrition and modern eating habits are inadequate to maintain adequate total body mass.

Myostatin is a polypeptide growth factor belonging to the superfamily of TGF-β. TGF-β has a large amount of isoforms, which are known to be involved in proliferation, apoptosis, differentiation, bone formation and maintenance of cells (Massague & Chen, 2000). Myostatin belongs to growth differentiation factor (GDF) No. 8, is involved in the growth and development of tissues, and acts by activating the Smad signaling system. In addition, the p21 gene has been reported to inhibit the proliferation of cell cycle and progenitor cells, thereby affecting bone formation and regeneration. Myostatin is produced mainly in skeletal muscle cells, causing muscle loss and muscle strength by the method of self-secretion. In addition, myostatin inhibits the expression of IGF-1 or Follistatin, which is involved in muscle hypertrophy, and causes protein synthesis and proliferation in myoblasts. It is known to suppress.

On the other hand, the seaweed grows about 10-30cm in height, and branches are split facing each other. The stem is dark green or yellow at first, but turns red in autumn, and is thick and fleshy like a small cactus. The leaves degenerate and are almost invisible and have a small piece of membranous scales. Three flowers bloom at upper nodes of branches from August to September. The skin is fleshy and has 1 to 2 stamens and 2 pistils. Fruits are enclosed in skins with pores and have black seeds. It is an annual herb that grows in groups near the tidal flats where seawater enters or in inland saltwater. In oriental medicine, it removes toxins and stool that accumulate in the body and has excellent treatment effect for various incurable diseases such as cancer, uterine fibroid, sinusitis, hypertension, hypotension, low back pain, diabetes, bronchial asthma, hypothyroidism, hyperthyroidism, skin disease, arthritis, etc. It is known to have.

Against this background, the present inventors have made diligent efforts to discover substances capable of treating myotropin by inhibiting myostatin expression causing muscle loss and muscle strength. The present invention has been completed by discovering that it can be used for the prevention or treatment of myopathy by inhibiting the increase.

It is an object of the present invention to provide a pharmaceutical composition for the prevention or treatment of myopathy, which contains a seaweed extract as an active ingredient.

Another object of the present invention relates to a food composition for preventing or improving myopathy, which contains a seaweed extract as an active ingredient.

However, the problem to be solved by the present invention is not limited to the above-mentioned problem, another task that is not mentioned will be clearly understood by those skilled in the art from the following description.

According to one embodiment of the present invention, there is provided a pharmaceutical composition for the prevention or treatment of myotropia, which contains a seaweed extract as an active ingredient.

According to one side, the seaweed extract may include laminarin (laminarin) or fucoidan (fucoidan).

According to one side, the pharmaceutical composition may inhibit the expression of myostatin mRNA or protein.

According to one side, the myostatin mRNA or protein expression may be due to bleomycin (bleomycin).

According to one side, the myostatin mRNA or protein expression may be due to UV.

According to one side, the pharmaceutical composition may inhibit the expression of MURF1 (Muscle RING-finger protein-1) mRNA or protein.

According to another embodiment of the present invention, there is provided a food composition for preventing or ameliorating myotropia, which contains seaweed as an active ingredient.

The pharmaceutical composition for the prevention or treatment of myotropia containing the persimmon of the present invention as an active ingredient inhibits the increase in the production of myostatin protein and mRNA expression which directly affect muscle loss and muscle strength, and thus, more fundamental It may have a prophylactic or therapeutic effect of myopathy.

It is to be understood that the effects of the present invention are not limited to the above effects, and include all effects deduced from the configuration of the invention described in the detailed description or claims of the present invention.

Figure 1 confirms the effect of inhibiting myostatin expression increased by UV during laminarin and fucoidan treatment.

Figure 2 confirms the effect of inhibiting the increased myostatin expression by bleomycin during laminarin and fucoidan treatment.

Figure 3 confirms the effect of inhibiting the expression of MURF1 (Muscle RING-finger protein-1) increased by UV during laminarin and fucoidan treatment.

Hereinafter, exemplary embodiments will be described in detail with reference to the accompanying drawings. However, various changes may be made to the embodiments so that the scope of the patent application is not limited or limited by these embodiments. It is to be understood that all changes, equivalents, and substitutes for the embodiments are included in the scope of rights.

The terminology used herein is for the purpose of description and should not be construed as limiting. Singular expressions include plural expressions unless the context clearly indicates otherwise. In this specification, terms such as "comprise" or "have" are intended to indicate that there is a feature, number, step, operation, component, part, or combination thereof described on the specification, and one or more other features. It is to be understood that the present invention does not exclude the possibility of the presence or the addition of numbers, steps, operations, components, components, or a combination thereof.

Unless defined otherwise, all terms used herein, including technical or scientific terms, have the same meaning as commonly understood by one of ordinary skill in the art. Terms such as those defined in the commonly used dictionaries should be construed as having meanings consistent with the meanings in the context of the related art, and shall not be construed in ideal or excessively formal meanings unless expressly defined in this application. Do not.

In addition, in the description with reference to the accompanying drawings, the same components will be given the same reference numerals regardless of the reference numerals and duplicate description thereof will be omitted. In the following description of the embodiment, if it is determined that the detailed description of the related known technology may unnecessarily obscure the gist of the embodiment, the detailed description thereof will be omitted.

According to one embodiment of the present invention, there is provided a pharmaceutical composition for the prevention or treatment of myotropia, which contains a seaweed extract as an active ingredient.

As used herein, the term " Salicorniaherbacea " is the name of the herbal medicine of Sungcorn, it means halophyte belonging to the family of Sukjumok.

As used herein, the term "extract" refers to an extract obtained by the extraction process of the plant, a diluent or concentrate of the extract, a dried product obtained by drying the extract, a crude or purified product of the extract, a mixture thereof, or the like, It includes the extract itself and extracts of all formulations that can be formed using the extract. The seaweed extract of the present invention may preferably be prepared and used in the form of a dry powder after extraction. In addition, the seaweed extract, can be extracted from the natural, hybrid or variety plants of the seaweed, can be extracted from plant tissue culture, and includes an extract extracted from the site of leaves, stems, roots, flowers, fruits and the like.

The seaweed extract may include laminarin (laminarin) or fucoidan (fucoidan).

As used herein, the term "laminarin" is a carbohydrate in the cell wall of brown algae, a hypotonic polysaccharide corresponding to starch of higher plants.

As used herein, the term "fucoidan" is a viscous polysaccharide contained in brown algae, a substance called fucose (Fucose) combined with a basic sugar and a sulfate group.

As used herein, the term "prevention" means any action that inhibits or delays the development of myopathy by administration of the composition.

As used herein, the term "treatment" means any action that improves or beneficially changes the symptoms of myotropia by administration of the composition.

Laminarin and fucoidan contained in the pharmaceutical composition of the present invention can inhibit myostatin mRNA or protein expression and MURF1 (Muscle RING-finger protein-1) protein expression, in particular bleomycin treatment or UV stimulation Increase in myostatin expression may be reduced (see FIGS. 1 to 3). The bleomycin is an anti-tumor antibiotic and is used as an effective anticancer agent for squamous cell carcinoma or malignant lymphoma, or is known as an aging-inducing factor.

The pharmaceutical composition of the present invention may be prepared by a method known in the pharmaceutical field for use as a medicament, and may be mixed with pharmaceutically acceptable carriers, excipients, diluents, stabilizers, preservatives, etc. to powders, granules, tablets, It may be prepared and used in the form of a capsule or injection. In addition, the composition may be prepared in a sustained release formulation including a base used for sustained release purposes in addition to the active ingredient so that the release of the active ingredient occurs slowly.

Pharmaceutically acceptable carriers may further include, for example, carriers for oral administration or carriers for parenteral administration. Carriers for oral administration may include lactose, starch, cellulose derivatives, magnesium stearate, stearic acid and the like. In addition, it may include various drug delivery materials used for oral administration to the peptide formulation. In addition, carriers for parenteral administration may include water, suitable oils, saline, aqueous glucose, glycols, and the like, and may further include stabilizers and preservatives. Suitable stabilizers include antioxidants such as sodium hydrogen sulfite, sodium sulfite or ascorbic acid. Suitable preservatives include benzalkonium chloride, methyl- or propyl-paraben and chlorobutanol. The pharmaceutical composition of the present invention may further include a lubricant, a humectant, a sweetener, a flavoring agent, an emulsifier, a suspension agent, and the like, in addition to the above components.

The diluent may be a non-aqueous solvent such as propylene glycol, polyethylene glycol, vegetable oils such as olive oil and peanut oil, or brine (preferably 0.8% saline), water containing a buffer medium (preferably 0.05M phosphate buffer) An aqueous solvent, such as these, etc. are mentioned, but it is not limited to this.

The excipients include starch, glucose, lactose, sucrose, gelatin, malt, rice, flour, chalk, silica gel, sodium stearate, glycerol monostearate, talc, sodium chloride, anhydrous skim milk, glycerol, propylene, glycol, water , Ethanol and the like, but is not limited thereto.

The stabilizer may include carbohydrates such as sorbitol, mannitol, starch, sucrose, dextran, glutamate, and glucose, and proteins such as animal, vegetable or microbial proteins such as milk powder, serum albumin and casein, but are not limited thereto. no.

The preservative may include thimerosal, merthiolate, gentamicin, neomycin, nistatin, amphotericin B, tetracycline, penicillin, streptomycin, polymyxin B, and the like, but is not limited thereto.

The pharmaceutical compositions of the invention can be administered to any mammal, including humans, by any means, for example orally or parenterally. Parenteral administration methods include intravenous, intramuscular, intraarterial, intramedullary, intradural intracardiac, transdermal, subcutaneous, intraperitoneal, intranasal, intestinal, topical, sublingual or intrarectal administration no.

The pharmaceutical composition of the present invention may be formulated into a preparation for oral or parenteral administration according to the route of administration as described above.

The total effective amount of the pharmaceutical composition of the present invention may be administered to a patient in a single dose, and may be administered by a divided therapeutic regimen administered in multiple doses for a long time. The pharmaceutical composition of the present invention may vary the content of the active ingredient depending on the extent of the disease. This may be determined in consideration of various factors such as the formulation method, the number of administration routes, as well as the age, weight, health condition, symptoms of the disease, administration time and method of the patient. In view of this, one of ordinary skill in the art will be able to determine appropriate effective dosages of the compositions of the present invention. The pharmaceutical composition according to the present invention is not particularly limited in terms of dosage form, route of administration and method of administration as long as the effect of the present invention is shown.

According to another embodiment of the present invention, there is provided a food composition for preventing or improving muscular dystrophy, which contains the persimmon extract as an active ingredient.

As used herein, the term “improvement” means any action that at least reduces the parameters associated with the condition being treated, for example, the extent of symptoms.

In addition, when the food composition of the present invention is used as a food additive, the composition may be added as it is or used with other food or food ingredients, and may be appropriately used according to a conventional method. Generally, in the preparation of food or beverages, the compositions of the invention are added in an amount of up to 15% by weight, preferably up to 10% by weight relative to the raw materials. However, in the case of long-term intake for the purpose of health and hygiene or for the purpose of health control, it may be below the above range, and since there is no problem in terms of safety, the active ingredient may be used in an amount above the above range.

There is no particular limitation on the kind of the food. Examples of the food to which the substance can be added include dairy products including meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, gum, ice cream, various soups, drinks, tea, drink, Alcoholic beverages and vitamin complexes, and the like and include all of the health foods in the conventional sense.

Hereinafter, the present invention will be described in more detail with reference to Examples. The following examples are described for the purpose of illustrating the present invention, but the scope of the present invention is not limited thereto.

Example 1 Verification of the Inhibitory Effect of Myotropia Induced by UV Irradiation

C2C12 muscle progenitor cells (ATCC, US) were cultured in DMEM containing 10% FBS. Cells were seeded with 3x10 ⁵ cells in a 60-pie dish. The next day after seeding was treated with laminarin and fucoidan and left for 1 hour. After 1 hour, the growth media (10% FBS DMEM) was removed and washed with PBS. After removing PBS, the UV cross-linker (UVP CL-1000) was irradiated with UV of 3J / m² with the plate cap removed (0 day). Incubation in a CO ₂ incubator for 24 hours, the growth media was removed in the same manner PBS wash and then irradiated with UV of 3J / m² (1 day).

The next day, harvested at the same time (2 days) and Western blot was used to measure the changes in the expression of myostatin and MURF1 proteins, muscle-modulating target proteins in cell lysates. Myostatin was analyzed using Anti-GDF8 (Genetex, GTX32624), MURF1 using anti-MURF1 antibody (Abcam, ab172479), and Beta-actin antibody (Enogene E12-041-4) as a protein loading control. It was.

RNA was isolated using RNeasy Mini Kit (Qiagen, US), and cDNA was synthesized using AMV Reverse Transcripatse (Promega, US), and then the degree of myostatin mRNA expression was confirmed by cDNA by PCR. PCR conditions were initially denatured at 95 ° C. for 5 minutes,

Denaturation at 95 ° C. for 5 seconds,

Annealing at 62.8 ° C. for 10 seconds and

The final extension was carried out at 72 ° C. for 10 minutes. Detection of mRNA expression was carried out using the following primers.

As a result, it was confirmed that the expression of myostatin and MURF1 increased by UV irradiation decreased as treated with laminarin and fucoidan, as shown in FIGS.

. Example 2 Verification of Myotropy Inhibitory Effect Induced by Bleomycin

C2C12 muscle progenitor cells (ATCC, US) were cultured in DMEM containing 10% FBS. At this time, the cells were seeded with 3x10 ⁵ cells in a 60-pie dish, and then left for 1 hour by bleomycin treatment after seeding.

Then, laminarin and fucoidan were treated and left for 1 hour, and growth media (10% FBS DMEM) was removed and washed with PBS. After washing, Western blot method was used to measure the expression changes of myostatin and MURF1, the muscle protein regulatory target proteins in cell lysates. The expression of myostatin was analyzed using Anti-GDF8 (Genetex, GTX32624) and the beta-actin antibody (Enogene E12-041-4) as a protein loading control.

RNA was isolated using RNeasy Mini Kit (Qiagen, US), and cDNA was synthesized using AMV Reverse Transcripatse (Promega, US), and then the degree of myostatin mRNA expression was confirmed by cDNA by PCR. PCR conditions were initially denatured at 95 ° C. for 5 minutes,

Denaturation at 95 ° C. for 5 seconds,

Annealing at 62.8 ° C. for 10 seconds and

The final extension was carried out at 72 ° C. for 10 minutes. mRNA expression was confirmed using the same primers as described in Example 1 above.

As a result, as shown in Fig. 1 and 2, it was confirmed that the expression of myostatin increased by the bleomycin treatment decreased with the treatment of laminarin and fucoidan.

The results of the above examples, it was found that the laminarin and fucoidan components of the seaweed extract can be usefully used for the prevention or treatment of muscular dystrophy by inhibiting the expression of myostatin and MURF1, which are indicator proteins related to muscle mass control. Can be.

Although the embodiments have been described with reference to the accompanying drawings, those skilled in the art may apply various technical modifications and variations based on the above. For example, the described techniques may be performed in a different order than the described method, and / or the described components may be combined or combined in a different form than the described method, or replaced or substituted by other components or equivalents. Appropriate results can be achieved.

Therefore, other implementations, other embodiments, and equivalents to the claims are within the scope of the following claims.

Claims (7)

1. A pharmaceutical composition for preventing or treating myotropenia, which contains the extract of Hamcho as an active ingredient.
2. The method of claim 1,

   The seaweed extract comprises laminarin (laminarin) or fucoidan (fucoidan), a pharmaceutical composition for the prevention or treatment of myopathy.
3. The method of claim 1,

   A pharmaceutical composition for preventing or treating myostatin, which inhibits myostatin mRNA or protein expression.
4. The method of claim 3,

   The myostatin mRNA or protein expression is due to bleomycin (bleomycin), a pharmaceutical composition for preventing or treating myopathy.
5. The method of claim 3,

   The myostatin mRNA or protein expression is due to UV, pharmaceutical composition for the prevention or treatment of myopathy.
6. The method of claim 1,

   MURF1 (Muscle RING-finger protein-1) A pharmaceutical composition for preventing or treating myopathy, which inhibits mRNA or protein expression.
7. Food composition for the prevention or improvement of myopathy, containing the seaweed extract as an active ingredient.

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