Leucine derivatives, compositions comprising the same, and applications thereof

Abstract

To provide cosmetic compositions, pharmaceutical compositions, quasi-drug compositions and food compositions with excellent skin improving effect, fatigue improving effect and / or menopausal symptom ameliorating effect in female.SOLUTION: A composition of the invention that contains glycyl-L-leucine peptide and / or L-leucyl-glycine peptide or a salt thereof as an effective ingredient exhibits an excellent effect in improvement of skin elasticity, skin moisturizing, prevention of skin aging, amelioration of fatigue, anti-inflammation, anti-oxidation and / or amelioration of female menopause symptoms. The composition of the invention containing glycyl-L-leucine peptide and / or L-leucyl-glycine peptide or salt thereof is, being safe for skin, usable as a cosmetic raw material, pharmaceutical ingredient, quasi-drug raw material or functional food material with excellent effect of interest, and can be contained as an index component or effective component.SELECTED DRAWING: Figure 1

Description

[Technical field]

[0001] The present invention relates to a leucine derivative, a composition containing the same, and uses thereof, and more particularly to novel cosmetic, pharmaceutical and food uses of a leucine derivative, as well as a cosmetic composition, a pharmaceutical composition, a quasi-drug composition and a food composition each containing the leucine derivative. [Background technology]

[0002] Aging is a life phenomenon, and all living organisms begin aging from birth. With improvements in sanitation and health care, human life expectancy has been significantly extended, and the proportion of the elderly population is steadily increasing (Kang Tae-jin, BioWave 2011, Vol. 13, No. 3(1)). Aging occurs throughout the body, and the overall physical function declines, as well as the physiological function of organs declines, as does lifestyle-related diseases such as cardiovascular disease and diabetes (Choi Hyun-cheol, Journal of the Korean Society of Dermatology, 1997; 35: 292-302).

[0003] The skin is the outermost organ of the body and serves various functions, such as preventing the loss of water, electrolytes, and proteins, as well as regulating body temperature, providing cutaneous sensation, and providing an immune system.

[0004] Like other organs, skin also ages as we get older. Skin aging can be divided into intrinsic skin aging and photoaging, and intrinsic skin aging is characterized by functional changes rather than visible morphological changes. Photoaging caused by chronic UV exposure overlaps with intrinsic aging and brings about significant structural and physiological changes, although it is not systemic or inevitable. Aged epidermis not only loses its inherent functions, such as the development of skin wrinkles, loss of barrier function, and immune abnormalities, but also has an aesthetic impact on appearance. Although skin aging cannot be stopped, various attempts are being made to prevent and delay skin aging.

[0005] Collagen is the main matrix protein produced by skin fibroblasts and exists in the extracellular matrix. Its important functions include mechanical toughness of the skin, resistance and binding of connective tissue, maintenance of cell adhesion, and induction of cell division and differentiation (during organism growth or wound healing). Collagen decreases with age and photoaging caused by exposure to ultraviolet rays, and collagen decrease is accelerated by collagenase enzyme activity, which breaks down collagen. This is known to be closely related to the formation of wrinkles in the skin.

[0006] Elastin is also a component of skin that is involved in skin elasticity and plays an important role in wrinkle formation. A sudden increase in the activity of elastase, an elastin-degrading enzyme, is known to be one of the causes of wrinkle formation. Elastase is the only enzyme that can degrade elastin, and it is known that inhibiting it can fundamentally reduce wrinkles.

[0007] Meanwhile, collagen and elastin fibers are matrix proteins that play an important role in retaining moisture in the dermis layer. These matrix proteins adsorb moisture and increase the moisture-retaining capacity of the structures they form, thereby enabling the skin to maintain an appropriate moisture content, and thus contributing to the maintenance of skin elasticity.

[0008] In addition, elastic fibers produced by dermal fibroblasts are a component of skin that is important in wrinkle formation, as they are involved in skin elasticity by cross-linking with collagen. The deficiency and decomposition of elastic fibers are known to be one of the main factors in the formation of wrinkles and loss of elasticity in the skin.

[0009] Currently, retinoids, adenosine, animal placenta-derived proteins, chlorella extract, etc. are known as wrinkle-improving cosmetics. The most widely known retinol is a substance that promotes collagen synthesis and inhibits elastase enzymes, but it is unstable and has safety issues such as irritation and redness when applied to the skin, so the amount of use is limited. In addition, chlorella extract, etc. are known to have little effect and are difficult to expect substantial improvement effects on skin wrinkles.

[0010] Therefore, there is an urgent need to develop an ingredient that is safe for the body, contains active ingredients that are safe and stable for the skin, and is more effective than substances currently known to have the effects of improving skin elasticity, moisturizing, or anti-aging.

[0011] Meanwhile, fatigue is one of the most common diseases in modern society and is a state of reduced physical performance (Uthayathas et al., 2007). Fatigue can be caused by excessive stress (Uthayathas et al., 2007) or abnormalities in immune function and antioxidant system (Cleare, 2003). Blood biomarkers such as blood urea nitrogen (BUN), alanine aminotransaminase (ALT), lactate dehydrogenase (LDH), and glucose are fatigue-related indicators (Klimas et al., 2012). LDH and creatine kinase (CK) levels are generally known as markers related to muscle damage (Coombes and McNaughton, 2000). Malate dehydrogenase (MDH) is also involved in the oxidative metabolism of fatigue (Callister et al., 2004). Fatigue is also induced by the occurrence of immune abnormalities in the oxidative stress pathway and various inflammatory pathways (Liu et al., 2000). After excessive exercise, reactive oxygen species are generated, which further exacerbates fatigue (Powers et al., 1999). To reduce such reactive oxygen species, superoxide dismutase (SOD) and catalase are produced (Liu et al., 2000). Nitric oxide (NO) is increased in patients with chronic fatigue (Suarez et al., 2010), and inflammatory cytokines are also related to fatigue symptoms (Liburt et al., 2010).

[0012] Currently, much research is being conducted into treating fatigue, but no suitable treatment has yet been found.

[0013] The causes of fatigue can be divided into a lack or inability to use energy sources stored in the body, an inflammatory reaction in the body due to metabolic action, abnormalities in the antioxidant system, accumulation of fatigue substances, and imbalance of homeostasis in the body, but it is known that most of the causes are due to the synergistic effects of many factors. In order to recover from such fatigue, it is necessary to provide a sufficient energy source, rest, and inhibit and remove the production of fatigue substances in the body, but in today's busy society, it is not easy to take in sufficient nutrition and rest, so there is a need to develop functional materials for fatigue recovery.

[0014] Meanwhile, menopause in women is the cessation of menstruation that occurs when the genetically determined lifespan of the ovaries, which lasts for about 50 years after birth, comes to an end, signifying the loss of reproductive ability and is a physiological change rather than a pathological phenomenon. The current average life expectancy of Korean women is 81.2 years (2011, Statistics Korea), and if we assume that the average age of menopause for Korean women, as determined by the Korean Society of Obstetrics and Gynecology, is 50 years, this means that women will spend more than one-third of their lives in a state of depletion of female hormones (Kim Seong-cheol, Pharmaceutical Information Institute).

[0015] When a woman reaches menopause, changes occur throughout the body, including the vascular system, musculoskeletal system, urogenital system, and cranial nerves, due to the imbalance and decrease in female hormone secretion. In other words, various diseases such as vasomotor symptoms and psychological symptoms such as facial flushing, night sweats, sleep disorders, fatigue, depression, anxiety, concentration disorders, memory disorders, pain during intercourse due to atrophy of the urogenital system, frequent urination, loss of skin elasticity due to collagen loss, sagging breasts, cardiovascular and musculoskeletal symptoms, and dementia are accompanied. Although menopausal symptoms vary from person to person, it has been reported that the more symptoms a woman experiences, the more severe the symptoms are, and the longer the period, the more the quality of life of the woman decreases, and menopausal symptoms are likely to progress to chronic diseases along with physical aging.

[0016] Menopausal symptoms can be treated by hormone therapy, drug therapy, exercise therapy, and diet therapy, but the female hormone therapy that is widely used medically increases the risk of developing diseases such as breast cancer, and if used for a long period of time, it may increase the incidence of uterine cancer, thrombotic vascular diseases, gallbladder diseases, and high blood pressure. Therefore, research is being continuously conducted into treatments that are safe for the human body and provide excellent therapeutic effects. [Prior art documents] [Non-patent literature]

[0017] [Non-Patent Document 1] Choi Hyun-cheol, Journal of the Korean Society of Dermatology, 1997;35:292-302 Summary of the Invention [Problem to be solved by the invention]

[0018] Therefore, the problem to be solved by the present invention is to solve the above problems and provide a new active ingredient that has few side effects and is safe for the human body, while having excellent effects of improving skin elasticity, moisturizing, or preventing skin aging, i.e., such a useful use of the active ingredient.

[0019] Another object of the present invention is to provide a new active ingredient which has few side effects and is safe for the human body while having excellent fatigue-improving, inflammation-improving or antioxidant effects, that is, to provide such useful uses of the active ingredient.

[0020] Another object of the present invention is to provide a new active ingredient having an excellent effect of improving menopausal symptoms, that is, to provide such a useful use of the active ingredient.

[0021] In other words, the problem to be solved by the present invention is to provide a composition containing the active ingredient having excellent efficacy, and further a composition containing the active ingredient having excellent efficacy as an active ingredient or an indicator ingredient. [Means for solving the problem]

[0022] In order to solve the above problems, the present invention provides a composition for improving skin elasticity, moisturizing skin, or preventing skin aging, preferably a cosmetic composition, pharmaceutical composition, quasi-drug composition, or food composition (preferably a functional food composition), which contains glycyl-L-leucine peptide and / or L-leucylglycine peptide; or a salt thereof as an active ingredient.

[0023] In other words, the present invention provides new uses of glycyl-L-leucine peptide and / or L-leucylglycine peptide; or a salt thereof, such as a cosmetic composition, a pharmaceutical composition, a quasi-drug composition or a food composition, as well as new uses for improving skin elasticity, moisturizing skin or preventing skin aging.

[0024] The present invention provides a method for improving skin elasticity, moisturizing skin, or preventing skin aging by applying glycyl-L-leucine peptide and / or L-leucylglycine peptide; or a salt thereof to the skin.

[0025] The glycyl-L-leucine peptide and / or the L-leucylglycine peptide can be an active ingredient or an indicator ingredient.

[0026] The inventors conducted extensive research to develop a peptide or a natural material containing the peptide that is effective in improving skin, and as a result, they confirmed that a composition containing glycyl-L-leucine peptide and / or L-leucylglycine as an active ingredient is effective in improving skin elasticity, moisturizing skin, and preventing skin aging, thereby completing the present invention.

[0027] The present inventors have experimentally confirmed that a composition containing glycyl-L-leucine and / or L-leucylglycine or a salt thereof as an active ingredient has a skin improving effect by promoting hyaluronic acid synthesis, promoting procollagen synthesis, and inhibiting elastase.

[0028] In the present invention, "improving skin elasticity" means reducing the degree of skin sagging or stretching. Also, the elasticity means maintaining the elasticity of the skin in a state where elastin and collagen are sufficiently present.

[0029] In the present invention, "skin moisturizing" means increasing the moisture of the skin and maintaining a moist state. The skin moisturizing effect also helps to improve skin wrinkles and increase elasticity.

[0030] In the present invention, "anti-aging of skin" means preventing and alleviating the occurrence of skin wrinkles, loss of elasticity, etc. As an example, skin aging can be suppressed by inhibiting the activity of elastase.

[0031] In another embodiment of the present invention, the present invention provides a composition for improving fatigue, improving inflammation or antioxidant properties, preferably a functional health food composition, a cosmetic composition, a pharmaceutical composition or a quasi-drug composition, comprising glycyl-L-leucine peptide and / or L-leucylglycine peptide; or a salt thereof as an active ingredient.

[0032] In other words, the present invention provides new uses of glycyl-L-leucine peptide and / or L-leucylglycine peptide; or salts thereof as functional health food compositions, cosmetic compositions, pharmaceutical compositions, or quasi-drug compositions, as well as new uses for improving fatigue, improving inflammation, or as an antioxidant.

[0033] The present invention provides a method for improving fatigue, inflammation or preventing oxidation by treating glycyl-L-leucine peptide and / or L-leucylglycine peptide; or a salt thereof.

[0034] The glycyl-L-leucine peptide and / or the L-leucylglycine peptide can be an active ingredient or an indicator ingredient.

[0035] The inventors experimentally confirmed that glycyl-L-leucine peptide and / or L-leucylglycine peptide; or a salt thereof prevents accumulation of lactate and creatine kinase (CK) in muscle in a mouse model with fatigue induced by exercise, and found that it is effective in improving fatigue, leading to the completion of the present invention. In addition, the inventors experimentally confirmed that the glycyl-L-leucine peptide and / or L-leucylglycine peptide; or a salt thereof reduces the secretion of tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) in spleen cells, thereby confirming an inflammation improving effect, and experimentally confirmed that the glycyl-L-leucine peptide and / or L-leucylglycine peptide; or a salt thereof increases the level of superoxide dismutase in blood, thereby confirming an antioxidant effect.

[0036] The term "improving fatigue" as used herein means to eliminate accumulated fatigue or to remove fatigue symptoms, and fatigue means a state in which the strength or sensitivity of cells, muscles, or organs is temporarily reduced due to physical or mental activity. If mental and physical activity is continued without being able to recover from fatigue, fatigue may accumulate and lead to overwork or chronic fatigue, which may lead to exercise disorders and even health disorders. Fatigue occurs due to excessive physical activity, and the physiological mechanism of fatigue is not limited to this, but it is due to the consumption of energy sources such as glucose and glycogen by physical activity. The higher the intensity of exercise, the faster muscle glycogen is consumed, and the lower the intensity, the less energy is consumed. However, even with low-intensity exercise, the consumption increases over time, causing fatigue. Another cause of fatigue is the accumulation of metabolic intermediates such as lactic acid and carbon dioxide in the body. Carbohydrates used as an energy source for the human body are broken down into glucose in the body, and the remaining portion is stored in the liver and muscles in the form of glycogen and used as an energy source when needed. When glycogen is broken down during exercise, if there is a sufficient supply of oxygen, it is completely broken down to generate a large amount of energy and sustain the exercise, but if there is insufficient oxygen supply, lactic acid is generated and accumulated. If lactic acid accumulates, chemical reactions do not proceed smoothly, causing fatigue. In the present invention, the prevention of the accumulation of lactic acid and creatine kinase (CK) in the muscles was experimentally confirmed in a mouse model in which fatigue was induced by exercise, and the fatigue-improving effect of the peptide was confirmed.

[0037] The term "improving inflammation" as used herein means not only suppressing inflammation but also removing and / or alleviating inflammation that has occurred. Here, inflammation is one of the defense responses of biological tissues to a certain stimulus, and refers to a complex lesion that simultaneously involves three types of symptoms: tissue degeneration, circulatory disorders and exudation, and tissue proliferation. More specifically, inflammation is part of innate immunity, and like other animals, human innate immunity recognizes cell surface patterns that are specifically present on pathogens. Phagocytes recognize cells with such surfaces as non-self and attack the pathogens. If pathogens break through the physical barriers of the body and invade, an inflammatory response occurs. The inflammatory response is an immunological defense mechanism that attempts to repair and regenerate the damaged area when the body is damaged by many factors, but if it occurs excessively or persistently, it induces acute or chronic inflammatory diseases. This can also be the cause of diseases such as circulatory disorders, multiple sclerosis, and Parkinson's disease. Inflammation occurs through many mechanisms in the body, including hormone secretion, cytokines, and C-reactive protein (CRP), and involves a large number of substances. Among them, various cytokines secreted by immune cells regulate the immune system, and some of them can induce inflammation. Therefore, the expression level of intracellular cytokines is an indicator of the activation of inflammatory responses. In the present invention, the peptide's inflammation-improving effect was confirmed experimentally by confirming that it reduced the secretion of tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) in spleen cells in a mouse model with fatigue induced by exercise.

[0038] The term "antioxidant" as used herein means to suppress oxidative stress. Substances that induce oxidative stress may be generated in the human body. When oxygen used as an energy source in the body is incompletely reduced, reactive oxygen species are released into the cytoplasm. Such reactive oxygen species damage surrounding cells and may further induce tissue damage due to the progression of inflammation. Such reactive oxygen species are generated under normal circumstances, but if the body does not have an antioxidant defense mechanism, human life cannot be sustained. Therefore, the human body has various forms of antioxidant defense systems to counter normal oxidative stress. However, if oxidative stress exceeds the normal level, the body will be seriously damaged, so measures against oxidative stress are necessary. Antioxidants are natural or synthetic substances that function to prevent or suppress oxidative stress. To evaluate the antioxidant power of these substances in the body, the content of antioxidants in the body, such as superoxide dismutase (SOD), catalase, peroxidase, and glutathione peroxidase (GPx), is evaluated by various methods. These substances mainly function to remove reactive oxygen species. In the present invention, the antioxidant effect of the peptide was confirmed by experimentally confirming that it increased the blood superoxide dismutase (SOD) level in a mouse model in which fatigue was induced by exercise.

[0039] In yet another embodiment of the present invention, the present invention provides a composition for alleviating, improving, or treating menopausal symptoms, preferably a functional health food composition, a cosmetic composition, a pharmaceutical composition, or a quasi-drug composition, comprising glycyl-L-leucine peptide and / or L-leucylglycine peptide; or a salt thereof as an active ingredient.

[0040] In other words, the present invention provides new uses of glycyl-L-leucine peptide and / or L-leucylglycine peptide; or salts thereof as functional health food compositions, cosmetic compositions, pharmaceutical compositions, or quasi-drug compositions, as well as for the alleviation, improvement, or treatment of menopausal symptoms.

[0041] The present invention provides a method for alleviating, ameliorating or treating menopausal symptoms by administering glycyl-L-leucine peptide and / or L-leucylglycine peptide; or a salt thereof.

[0042] The glycyl-L-leucine peptide and / or the L-leucylglycine peptide can be an active ingredient or an indicator ingredient.

[0043] In the present invention, the term "menopausal symptoms" refers to symptoms and diseases that appear in women before and after menopause due to a decrease in estrogen secretion caused by ovarian aging, etc. It is also called "menopausal syndrome" or "menopausal symptoms". Examples of menopausal or menopausal symptoms include, but are not limited to, facial flushing, sweating, nervousness, depression, dizziness, fatigue, joint pain, muscle pain, headache, chest palpitations, feeling of formication, sweating during sleep, sleep disorders, dry skin, vaginal dryness, vaginal atrophy, lower urethral atrophy, pain during intercourse, vaginitis, cystitis, pain during urination, urgent urination, concentration disorders, memory disorders, anxiety, irritability, memory decline, and osteoporosis. In addition, cardiovascular disorders such as heart disease, high blood pressure, and stroke are also included.

[0044] In the present invention, the term "prevention" refers to any action of administering the composition of the present invention to suppress or delay the target symptoms.

[0045] In the present invention, the term "treatment" refers to any action of administering the composition of the present invention to improve or cure a target symptom or disease.

[0046] In the present invention, "amelioration" refers to any action of administering the composition of the present invention to improve or favorably change the target symptoms compared to before administration.

[0047] In order to solve the above problems and achieve the object of the present invention, the present invention provides a composition comprising a compound represented by the following Chemical Formula 1 and / or a compound represented by the following Chemical Formula 2; or a salt thereof.

[0048] The compound represented by the following Chemical Formula 1 and / or the compound represented by the following Chemical Formula 2 may be an active ingredient or an indicator ingredient.

[0049] [ka]

[0050] The compound represented by Chemical Formula 1 has the molecular formula CH 16 N2O3 has a molecular weight of 188.22 and is called glycyl-L-leucine. It is a dipeptide composed of two amino acids, glycine and L-leucine, and is abbreviated as Gly-Leu or GL.

[0051] The present invention is not particularly limited by the method for obtaining the glycyl-L-leucine, and it may be derived from a natural product, chemically synthesized by a method known in the art to which the present invention pertains, or a commercially available substance may be used. Preferably, a substance extracted from the placenta of a mammal such as a pig, cow, sheep, or horse may be used, and more preferably, a substance extracted from the placenta of a pig may be used.

[0052] [ka]

[0053] The compound represented by Chemical Formula 2 has the molecular formula CH 16 N2O3 has a molecular weight of 188.22 and is called L-leucylglycine. It is a dipeptide composed of two amino acids, L-leucine and glycine, and is abbreviated as Leu-Gly or LG.

[0054] The present invention is not particularly limited by the method for obtaining the L-leucylglycine, and it may be derived from a natural product, chemically synthesized by a method known in the art to which the present invention belongs, or a commercially available substance may be used. Preferably, a substance extracted from the placenta of a mammal such as a pig, cow, sheep, or horse may be used, and more preferably, a substance extracted from the placenta of a pig may be used.

[0055] In the cosmetic composition, pharmaceutical composition, quasi-pharmaceutical composition or food composition according to the present invention, the content of the compound represented by Chemical Formula 1 and / or the compound represented by Chemical Formula 2 or a salt thereof is 0.00000001 to 100 wt % based on the total weight of the cosmetic composition, pharmaceutical composition, quasi-pharmaceutical composition or food composition, that is, the compound represented by Chemical Formula 1 and / or the compound represented by Chemical Formula 2 can be ingested or applied as it is.

[0056] Preferably, in the present invention, when the active ingredient is a mixture of the compound represented by Chemical Formula 1 and the compound represented by Chemical Formula 2, the components may be mixed in a weight ratio of 1:0.05 to 20, preferably 1:0.1 to 10. The present inventors have experimentally confirmed the effects of each component as well as a synergistic effect when the active ingredients are mixed in a weight ratio of 1:1 within the above range.

[0057] In the present invention, the compound represented by Chemical Formula 1 and / or the compound represented by Chemical Formula 2 used as an active ingredient can be used in the form of a salt. In this case, if the composition according to the present invention is a food, it can be used in the form of a salt that is not harmful to health and exhibits the intended effect of the present invention, if it is a cosmetic composition, it can be used in the form of a cosmetically acceptable salt, and if it is a pharmaceutical or quasi-drug composition, it can be used in the form of a pharmaceutical acceptable salt. The salt of the compound represented by Chemical Formula 1 and / or the compound represented by Chemical Formula 2 is a salt prepared by adding an acid (e.g., a non-toxic or less toxic organic acid or inorganic acid) or a base to the compound represented by Chemical Formula 1 and / or the compound represented by Chemical Formula 2.

[0058] Examples of the salts include, but are not limited to, sodium salts, salicylates, benzoates, calcium salts, citrates, sulfites and bisulfites, aluminum salts, hydrochlorides, permanganates, potassium salts, acetates, bromates, nitrates, carbonates, bicarbonates, thiocyanates, ferrous salts, thiosulfates, ferric salts, cupric salts, lactates, magnesium salts, sulfates, phosphates, bromides, zinc salts, iodides, and succinates.

[0059] The compound represented by Chemical Formula 1 and / or the compound represented by Chemical Formula 2 of the present invention; or a salt thereof can exist in a non-solvated form as well as a solvated form including a hydrate, an ethanol solvate, etc. The compound represented by Chemical Formula 1 and / or the compound represented by Chemical Formula 2 of the present invention; or a salt thereof can exist in a crystalline or amorphous form, and all such physical forms are within the scope of the present invention.

[0060] The present invention provides a food composition for improving skin elasticity, moisturizing skin, or preventing skin aging, comprising a compound represented by Chemical Formula 1 and / or a compound represented by Chemical Formula 2; or a salt thereof as an active ingredient.

[0061] The present invention provides a food composition for alleviating fatigue, alleviating inflammation or providing antioxidant properties, comprising a compound represented by Chemical Formula 1 and / or a compound represented by Chemical Formula 2; or a salt thereof as an active ingredient.

[0062] The present invention provides a food composition for improving, preventing or treating menopausal symptoms, comprising a compound represented by Chemical Formula 1 and / or a compound represented by Chemical Formula 2 or a salt thereof as an active ingredient. Preferably, the food composition may be a functional food.

[0063] The term "functional food" as used herein means a food product obtained by adding the compound represented by Formula 1 and / or the compound represented by Formula 2 to food materials such as beverages, teas, spices, gums, confectioneries, etc., or by encapsulating, powdering, or suspending the same, which has a specific health effect when ingested, and which has the advantage of being free of side effects that may occur when taking medicines for a long period of time, since it is made from food materials, unlike general medicines. The functional food of the present invention thus obtained can be ingested daily, and is therefore very useful, as it is expected to have excellent effects on improving skin elasticity, moisturizing skin or preventing skin aging, relieving fatigue, relieving inflammation, and alleviating, preventing, or treating antioxidant and / or menopausal symptoms.

[0064] When the compound represented by Chemical Formula 1 and / or the compound represented by Chemical Formula 2 or a salt thereof is used as a food additive, the compound represented by Chemical Formula 1 and / or the compound represented by Chemical Formula 2 or a salt thereof may be added as is or may be used together with other foods or food ingredients, and may be used appropriately by a conventional method. The amount of the active ingredient to be mixed may be appropriately determined depending on the purpose of use (prevention, health, or therapeutic treatment). In general, when producing a food or beverage, the composition of the present invention is added in an amount of 15 parts by weight or less, preferably 10 parts by weight or less, based on the raw material. However, in the case of long-term intake for the purpose of health and hygiene or health regulation, the amount may be within the above range, and since there is no problem in terms of safety, the active ingredient may be used in an amount greater than the above range. There is no particular limitation on the type of the food. Examples of foods to which the substance can be added include meats, sausages, breads, chocolates, candies, snacks, confectionery, pizza, ramen and other noodles, gums, dairy products including ice cream, various soups, beverages, teas, energy drinks, alcoholic beverages and vitamin complexes, including all foods in the usual sense.

[0065] When the food is a beverage, the content may be 1 to 30 g, preferably 3 to 20 g, based on 100 ml. The composition may also contain additional ingredients that are commonly used in food compositions and can improve smell, taste, vision, etc. For example, vitamins A, C, D, E, B1, B2, B6, B12, niacin, biotin, folate, panthotenic acid, etc. may also be included. Minerals such as zinc (Zn), iron (Fe), calcium (Ca), chromium (Cr), magnesium (Mg), manganese (Mn), copper (Cu), etc. may also be included. Amino acids such as lysine, tryptophan, cysteine, and valine may also be included. In addition, food additives such as preservatives (potassium sorbate, sodium benzoate, salicylic acid, sodium dehydroacetate, etc.), bactericides (bleaching powder, high bleaching powder, sodium hypochlorite, etc.), antioxidants (butyl hydroxyanisole (BHA), butyl hydroxytoluene (BHT), etc.), colorants (tar dyes, etc.), color formers (sodium nitrite, sodium nitrite, etc.), bleaching agents (sodium sulfite), seasonings (MSG (monosodium glutamate), etc.), sweeteners (dulcin, cyclamate, saccharin, sodium, etc.), flavors (vanillin, lactones, etc.), leavening agents (alum, potassium D-hydrogen tartrate, etc.), strengtheners, emulsifiers, thickeners (thickening agents), coating agents, gum base agents, foam inhibitors, solvents, and improvers can be added. The additives are selected according to the type of food and used in appropriate amounts. The ratio of such additives is not particularly important, but is generally selected within the range of 0.01 to 0.8 parts by weight per 100 parts by weight of the composition of the present invention.

[0066] The food of the present invention can be produced by a method commonly used in the art, and can be produced by adding raw materials and ingredients commonly added in the art. In addition, unlike general medicines, since the food is made from raw materials, it has the advantage of being free of side effects that may occur when taking medicines for a long period of time, and is highly portable.

[0067] In another aspect, there is provided a cosmetic composition for improving skin elasticity, moisturizing skin, or preventing skin aging, comprising a compound represented by Chemical Formula 1 and / or a compound represented by Chemical Formula 2; or a cosmetically acceptable salt thereof as an active ingredient.

[0068] In another aspect, there is provided a cosmetic composition for alleviating fatigue, alleviating inflammation or providing antioxidant properties, comprising a compound represented by Chemical Formula 1 and / or a compound represented by Chemical Formula 2; or a salt thereof as an active ingredient.

[0069] In another aspect, there is provided a cosmetic composition for improving, preventing or treating menopausal symptoms, comprising a compound represented by Chemical Formula 1 and / or a compound represented by Chemical Formula 2; or a salt thereof as an active ingredient.

[0070] Cosmetics containing the composition as an active ingredient of the cosmetic composition can be prepared in the form of a general emulsion or solubilization formulation, for example, a toner such as a softening toner or a nutritious toner, a milky lotion such as a facial lotion or a body lotion, a cream such as a nutritious cream, a moisture cream, or an eye cream, an essence, a cosmetic ointment, a spray, a gel, a pack, a sunscreen, a makeup base, a liquid type, a solid type, or a spray type foundation, a powder, a makeup remover such as a cleansing cream, a cleansing lotion, or a cleansing oil, a cleansing foam, a soap, a body wash, or the like, but are not limited thereto.

[0071] When the composition is commercialized as a cosmetic product, in the case of wash-off type cosmetics such as makeup removers and cleansers in which the active ingredient remains on the skin for a short time, the composition of the present invention may be contained in a relatively high concentration, whereas in the case of leave-on type cosmetics such as lotions, milks, creams, essences, in which the active ingredient remains on the skin for a long time, the composition of the present invention may be contained in a lower concentration than in wash-off type cosmetics.

[0072] In addition, the cosmetic composition of the present invention may further comprise auxiliary agents commonly used in the field of cosmetology, such as fatty substances, organic solvents, solubilizers, thickening and gelling agents, emollients, antioxidants, suspending agents, stabilizers, foaming agents, fragrances, surfactants, water, ionic or non-ionic emulsifiers, fillers, sequestering and chelating agents, preservatives, vitamins, blocking agents, moisturizing agents, essential oils, dyes, pigments, hydrophilic or lipophilic active agents, lipid vesicles, or any other ingredients commonly used in cosmetics.

[0073] The cosmetic composition of the present invention may further contain one or more cosmetically acceptable carriers that are commonly incorporated into skin cosmetics.

[0074] The cosmetically acceptable carrier contained in the cosmetic composition of the present invention varies depending on the dosage form. When the dosage form of the present invention is an ointment, paste, cream or gel, the carrier component can be animal oil, vegetable oil, wax, paraffin, starch, tragacanth, cellulose derivative, polyethylene glycol, silicone, bentonite, silica, talc, zinc oxide or a mixture thereof.

[0075] When the dosage form of the present invention is a powder or spray, lactose, talc, silica, aluminum hydroxide, calcium silicate, polyamide powder or mixtures thereof can be used as the carrier component, and especially when it is a spray, it can further contain a propellant such as hydrochlorofluorocarbon, propane / butane or dimethyl ether.

[0076] When the dosage form of the present invention is a solution or emulsion, a solvent, solubilizer, or emulsifier is used as a carrier component, such as water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl benzoate, propylene glycol, 1,3-butyl glycol oil, in particular cottonseed oil, peanut oil, corn germ oil, olive oil, castor oil, and sesame oil, glycerin fatty acid esters, polyethylene glycol, or fatty acid esters of sorbitan.

[0077] When the dosage form of the present invention is a suspension, the carrier component may be a liquid diluent such as water, ethanol or propylene glycol, a suspending agent such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester and polyoxyethylene sorbitan ester, microcrystalline cellulose, aluminum metahydroxide, bentonite, agar or tragacanth.

[0078] When the dosage form of the present invention is a soap, alkali metal salts of fatty acids, fatty acid hemiester salts, fatty acid protein hydrolysates, isethionates, lanolin derivatives, fatty alcohols, vegetable oils, glycerol, sugars, etc. can be used as carrier components.

[0079] In another aspect, the present invention provides a pharmaceutical composition for improving skin elasticity, moisturizing skin, or preventing skin aging, comprising a compound represented by Chemical Formula 1 and / or a compound represented by Chemical Formula 2; or a pharma- ceutical acceptable salt thereof, as an active ingredient.

[0080] In another aspect, the present invention provides a pharmaceutical composition for improving fatigue, improving inflammation, or antioxidant properties, comprising a compound represented by Chemical Formula 1 and / or a compound represented by Chemical Formula 2; or a salt thereof as an active ingredient.

[0081] In another aspect, the present invention provides a pharmaceutical composition for improving, preventing or treating menopausal symptoms, comprising a compound represented by Chemical Formula 1 and / or a compound represented by Chemical Formula 2; or a salt thereof, as an active ingredient.

[0082] The composition may have any one of the dosage forms selected from the group including a solution, a gel, an emulsion, a suspension, a microemulsion, a microcapsule, a liposome, a cream, a lotion, an ointment, an aerosol, a spray, a paste, and a patch.

[0083] The pharmaceutical compositions of the present invention may be used in a single formulation or may be prepared and used in a combined formulation by further including a drug recognized to have anti-inflammatory or antioxidant effects, and may be prepared in unit dose form or in multi-dose containers by formulating with a pharma- ceutically acceptable carrier or excipient.

[0084] The term "pharmaceutical acceptable carrier" as used herein means a carrier or diluent that does not stimulate the organism and does not inhibit the biological activity and properties of the compound to be injected. The type of the carrier that can be used in the present invention is not particularly limited, and any pharmaceutical acceptable carrier that is commonly used in the art can be used. Non-limiting examples of the carrier include saline, sterile water, Ringer's solution, buffered saline, albumin injection solution, dextrose solution, maltodextrin solution, glycerol, ethanol, etc. These can be used alone or in combination of two or more. The carrier can include a non-naturally occurring carrier.

[0085] If necessary, other common additives such as antioxidants, buffers and / or bacteriostatic agents may be added and used, and diluents, dispersants, surfactants, binders, lubricants and the like may be further added to formulate the composition into injection forms such as aqueous solutions, suspensions, emulsions, pills, capsules, granules, tablets, and the like.

[0086] In addition, the pharmaceutical composition of the present invention may contain a pharma- ceutical effective amount of the compound represented by Chemical Formula 1 and / or the compound represented by Chemical Formula 2; or a pharma- ceutical acceptable salt thereof. As used herein, the term "pharma- ceutical effective amount" means an amount sufficient to improve skin elasticity, moisturize skin, or prevent skin aging, or an amount sufficient to improve fatigue, improve inflammation, or induce an antioxidant effect, or an amount sufficient to alleviate, improve, or treat menopausal symptoms, at a reasonable therapeutic benefit / risk ratio applicable to medical treatment, and may be administered in an amount of generally 0.001 to 1000 mg / kg, preferably 0.005 to 200 mg / kg, more preferably 0.01 to 100 mg / kg, once or several times a day. However, for purposes of the present invention, it is desirable to vary and apply a specific therapeutically effective amount for a particular patient depending on various factors such as the type and degree of response to be achieved, the specific composition including the use of other preparations, if applicable, the age, weight, general health condition, sex and diet of the patient, the administration time, administration route and excretion rate of the composition, the treatment period, drugs used together with or simultaneously with the specific composition, and similar factors well known in the pharmaceutical art.

[0087] In another aspect, the present invention provides a quasi-drug composition for improving skin elasticity, moisturizing skin, or preventing skin aging, comprising a compound represented by Chemical Formula 1 and / or a compound represented by Chemical Formula 2; or a pharma-ceutical salt thereof, as an active ingredient.

[0088] In another aspect, the present invention provides a quasi-drug composition for improving fatigue, improving inflammation or providing antioxidant properties, comprising a compound represented by Chemical Formula 1 and / or a compound represented by Chemical Formula 2; or a pharma-ceutical acceptable salt thereof as an active ingredient.

[0089] In another aspect, the present invention provides a quasi-pharmaceutical composition for alleviating, improving or treating menopausal symptoms, comprising a compound represented by Chemical Formula 1 and / or a compound represented by Chemical Formula 2; or a pharma-ceutical acceptable salt thereof, as an active ingredient.

[0090] The term "quasi-drug" as used herein means a fiber, rubber product or similar used for the purpose of treating, mitigating, treating or preventing a disease in humans or animals, a non-instrument or machine or similar that has a weak or no direct effect on the human body, or a preparation used for sterilization, insecticide or similar purposes for infection prevention, excluding a non-instrument, machine or device among items used for the purpose of diagnosing, treating, mitigating, treating or preventing a disease in humans or animals, and a non-instrument, machine or device among items used for the purpose of pharmacologically affecting the structure and function of humans or animals. The quasi-drug may also include skin topical agents and personal hygiene products. Preferably, the quasi-drug may be a disinfectant, shower foam, gargle, wet tissue, detergent, soap, hand wash or ointment, but is not limited thereto.

[0091] When the composition according to the present invention is used as a quasi-drug additive, the composition can be added as it is or can be used together with other quasi-drugs or quasi-drug ingredients, and can be used appropriately by a conventional method. The amount of the active ingredient to be mixed can be suitably determined according to the purpose of use.

[0092] The present invention also provides a composition for improving skin elasticity, moisturizing skin, and preventing skin aging, preferably a cosmetic composition, pharmaceutical composition, quasi-drug composition, or food composition, which contains glycyl-L-leucine peptide and / or L-leucylglycine peptide; or a salt thereof as an indicator component.

[0093] In another aspect, the present invention provides a quasi-drug composition for improving fatigue, improving inflammation or providing antioxidant properties, comprising a compound represented by Chemical Formula 1 and / or a compound represented by Chemical Formula 2; or a pharma-ceutical acceptable salt thereof as an index component.

[0094] In another aspect, the present invention provides a quasi-pharmaceutical composition for alleviating, improving or treating menopausal symptoms, comprising a compound represented by Chemical Formula 1 and / or a compound represented by Chemical Formula 2; or a pharma-ceutical acceptable salt thereof as an index component. Effect of the Invention

[0095] The composition containing the glycyl-L-leucine peptide and / or L-leucylglycine peptide or a salt thereof according to the present invention as an active ingredient exhibits excellent effects in improving skin elasticity, moisturizing skin, and preventing skin aging.

[0096] The composition containing the glycyl-L-leucine peptide and / or L-leucylglycine peptide according to the present invention or a salt thereof exhibits excellent effects in alleviating fatigue, alleviating inflammation, or antioxidation.

[0097] The composition containing the glycyl-L-leucine peptide and / or L-leucylglycine peptide according to the present invention or a salt thereof is highly effective in alleviating menopausal symptoms.

[0098] The composition containing glycyl-L-leucine peptide and / or L-leucylglycine peptide or a salt thereof according to the present invention can be used as a cosmetic ingredient, pharmaceutical ingredient, quasi-drug ingredient, or food ingredient that is safe for the skin and has excellent effects of improving skin conditions.

[0099] The composition containing the glycyl-L-leucine peptide and / or L-leucylglycine peptide according to the present invention or a salt thereof is safe and can be used as a cosmetic ingredient, pharmaceutical ingredient, functional health food ingredient, or feed ingredient, which has excellent fatigue-relieving, inflammation-relieving, or antioxidant effects.

[0100] The composition containing glycyl-L-leucine peptide and / or L-leucylglycine peptide or a salt thereof according to the present invention can be used as a cosmetic ingredient, pharmaceutical ingredient, functional health food ingredient, or feed ingredient, which is safe yet has excellent effects in alleviating menopausal symptoms. [Brief description of the drawings]

[0101] [Figure 1] 1 is a graph showing the hyaluronic acid synthesis promoting effect of glycyl-L-leucine peptide and / or L-leucylglycine peptide. [Diagram 2] 1 is a graph showing the procollagen synthesis promoting effect of glycyl-L-leucine peptide and / or L-leucylglycine peptide. [Diagram 3] 1 is a graph showing the elastase inhibitory activity of glycyl-L-leucine peptide and / or L-leucylglycine peptide. [Figure 4] 1 is a graph showing the effect of alleviating inflammation and fatigue by suppressing excessive prostaglandin E2 (PGE2) secretion according to an example of the present invention and a comparative example. [Diagram 5] 1 is a graph showing the antioxidant effect based on DPPH free radical scavenging activity according to examples of the present invention and comparative examples. [Figure 6] 1 is a graph showing the effect of improving fatigue by preventing intramuscular lactic acid accumulation according to an embodiment of the present invention and a comparative example. [Figure 7] 1 is a graph showing the effect of improving fatigue by preventing intramuscular creatine kinase (CK) accumulation according to an embodiment of the present invention and a comparative example. [Figure 8] 1 is a graph showing the inflammation-improving effect due to the reduction of blood tumor necrosis factor-α (TNF-α) according to an embodiment of the present invention and a comparative example. [Figure 9] 1 is a graph showing the inflammation-improving effect due to the reduction of interleukin-6 (IL-6) in blood according to an example of the present invention and a comparative example. [Figure 10] 1 is a graph showing the antioxidant effect by increasing blood superoxide dismutase (SOD) according to an embodiment of the present invention and a comparative example. [Figure 11]The experimental results show that the estrogen receptor activity is increased by treating the examples and comparative examples of the present invention. It was confirmed that the estrogen receptor activity was significantly increased when glycyl-L-leucine peptide and L-leucylglycine peptide were treated together. From these results, it was confirmed that the glycyl-L-leucine peptide and / or L-leucylglycine peptide of the present invention can be effectively used for the treatment, improvement, and / or prevention of menopausal disorders. [Figure 12] The results show that the glycyl-L-leucine peptide and / or L-leucylglycine peptide of the present invention inhibits osteoclast differentiation and can be effectively used to treat, improve, and / or prevent postmenopausal osteoporosis. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0102] The present invention will be described in detail below with reference to examples to aid in understanding the present invention. However, the examples according to the present invention can be modified into many other forms, and the scope of the present invention should not be construed as being limited to the following examples. The examples of the present invention are provided to more completely explain the present invention to those with average knowledge in the art.

[0103] Reference Example 1: Glycyl-L-leucine peptide substance information [ka] Substance name: Glycyl-L-Leucine CAS No.:869-19-2 Molecular formula: C8H 16 N2O3 Molecular weight: 188.22 Purchased from: Bachem AG (Bubendorf, Switzerland)

[0104] Reference Example 2: Substance information for L-leucylglycine peptide [ka] Substance name: L-Leucylglycine CAS No.:686-50-0 Molecular formula: C8H 16 N2O3 Molecular weight: 188.22 Purchased from: Bachem AG (Bubendorf, Switzerland)

[0105] Experimental Example 1: Promotion effect of hyaluronic acid synthesis As skin aging progresses, collagen and elastin, which are made up of fibrous tissue that act as pillars to support the structure of the skin, harden and become insoluble. The matrix is ​​a substance that fills the spaces between cells and between fibrous substances such as collagen and elastin. The matrix has a high water retention capacity, and representative examples of such matrices include hyaluronic acid and mucopolysaccharides. Hyaluronic acid is a polysaccharide that is widely distributed in intracellular tissues such as synovial fluid and skin. It is a polymer of hyaluronan, which is a combination of N-acetylglucosamine and glucuronic acid, and has moisturizing and viscous properties. If these substances decrease with the progression of aging, dry skin is induced and the elasticity of the skin decreases (Kim Yun-beom, Journal of Dong-i Medical Physiological Pathology, 2010;24(4):533~542).

[0106] In order to confirm the effect of glycyl-L-leucine peptide or L-leucylglycine peptide on hyaluronic acid synthesis, the inventors cultured normal human epidermal keratinocytes and then treated the samples. After 24 hours, the culture medium of the cultured cells was removed, and the amount of hyaluronic acid produced in the culture medium was measured using Hyaluronan DuoSet ELISA Kit (DY3614) and normalized for protein content. Retinoic acid was used as a positive control. The untreated control group was set as 100% and the test results are shown in Figure 1.

[0107] As a result, it was confirmed that glycyl-L-leucine peptide or L-leucylglycine peptide increases the production of hyaluronic acid even at low concentrations, which means that it helps moisturize the skin and improves the condition of the skin. It was also confirmed that the production of hyaluronic acid increased when glycyl-L-leucine and L-leucylglycine were treated together compared to when they were treated alone.

[0108] Experimental Example 2: Procollagen synthesis promotion effect Procollagen is a component of collagen, synthesized in fibroblasts and osteoblasts, and is finally converted into collagen by procollagen peptase. Increased procollagen is related to the binding strength and elasticity of the skin.

[0109] To confirm the effect of glycyl-L-leucine peptide or L-leucylglycine peptide in promoting procollagen synthesis, normal human primary dermal fibroblasts-neonatal were cultured and then the samples were treated. After 24 hours, the culture medium for the cultured cells was removed and the amount of procollagen was measured using a Procollagen type IC peptide (PIP) kit. The cells attached to the bottom were washed with DPBS and dissolved in 1N NaOH, and the total protein amount was measured using a BCA Protein Assay Kit to determine the amount of procollagen synthesis per protein. TGF-β was used as a positive control. The untreated control group was set as 100% and the test results are shown in Figure 2.

[0110] As a result, it was confirmed that glycyl-L-leucine and L-leucylglycine peptides increase procollagen synthesis, which is beneficial for skin elasticity. It was also confirmed that the synthesis of procollagen was increased when glycyl-L-leucine and L-leucylglycine were treated together more than when they were treated alone.

[0111] Experimental Example 3: Elastase inhibitory activity In the dermis tissue of the skin, collagen and elastin form a network structure to maintain the elasticity of the skin, but elasticity and moisture decrease due to internal and external stress such as age and ultraviolet rays, and the network structure of elastin collapses due to overexpression of elastase, causing the skin to sag and wrinkle, and skin aging progresses. Therefore, skin aging can be suppressed by inhibiting the activity of elastase, which is one of the main causes of skin aging and is an enzyme that breaks down elastin (Lee Jaewon, Journal of the Korean Society of Food and Nutrition, 2008, 21(2);143~147).

[0112] The sample and 1U / ml porcine pancreatic elastase (Sigma, E1250) dissolved in 50mM Tris-HCl buffer (pH 8.0) were added to a 96-well plate, and the mixture was incubated at room temperature for 30 minutes. After adding N-succinyl-(L-Ala)3-p-nitroanilide as a substrate, the amount of p-nitroanilide produced from the substrate was measured at 410nm. The test results are shown in Figure 3, based on the untreated control group.

[0113] As a result, it was confirmed that glycyl-L-leucine peptide, L-leucylglycine peptide, and a mixture of glycyl-L-leucine peptide and L-leucylglycine peptide (1:1) inhibit elastase activity, and that skin aging can be suppressed by inhibiting the activity of elastase, which is the main cause of skin aging.

[0114] Experimental example 4: Test tube experiment The present inventors conducted test-tube experiments prior to evaluation of the fatigue improvement, inflammation improvement or antioxidant effects, and female menopausal symptom improvement effects of a composition containing glycyl-L-leucine and / or L-leucylglycine. Glycyl-L-leucine or L-leucylglycine was provided by Bachem AG (Bubendorf, Switzerland). As shown in Table 1 below, a composite composition was prepared and used in the experiment.

[0115] [Table 1]

[0116] Experimental Example 5: Inhibition of excessive prostaglandin E2 (PGE2) secretion reduces inflammation and fatigue Cytokines are proteins secreted mainly by monocytes, lymphocytes, T cells, B cells, natural killer cells, and fibroblasts during inflammatory reactions, and act on cell membrane receptors like neurotransmitters, or on intracellular receptors like hormones to transmit information between cells. Inflammatory cytokines, such as interleukin-1β, -2, -6 (IL-1β, -2, -6), IFN-γ, and TNF-α, activate cyclooxygenase-2 (COX-2) to increase prostaglandin E2 (PGE2), activating inflammatory cells and inducing the inflammatory process (Song Hee-rim et al., Journal of the Korean Society of Psychopharmaceutical Science 2013;24:5-10). The present inventors evaluated the anti-inflammatory effect of inhibiting prostaglandin E2 (PGE2) secretion.

[0117] First, to measure prostaglandin E2 (PGE2), mouse macrophage-like cell line RAW264.7 (KCLB no. 40071, Korea Cell Line Bank, Seoul, Korea) was purchased and 2.5 × 10 6The cells were then dispensed into a 96-well microplate at a cell concentration of 100 μl / ml. The medium used was DMEM (GibcoBRL, Grand Island, NY, USA) supplemented with 10% FBS (GibcoBRL, Grand Island, NY, USA) and 1% penicillin / streptomycin (Thermo Scientific Hyclone, Waltham, MA, USA), and cultured in an environment of 5% CO2 and 37°C. The cell culture medium into which the RAW264.7 cells were dispensed was treated with 1 μg / ml of lipopolysaccharide (LPS, Sigma, St. Louis, MO, USA) to induce inflammation, and then treated according to the above Experimental Example 1. The cells were then cultured for 24 hours, centrifuged at 900 rpm and 4°C for 5 minutes, and the cell culture medium was collected to confirm the amount of prostaglandin E2 (PGE2) secreted in the culture medium. Prostaglandin E2 (PGE2) was measured using a commercially available PGE2 ELISA kit (MyBioSource Co., Ltd., San Diego, CA, USA) according to the manufacturer's instructions.

[0118] As a result, as shown in Figure 4, prostaglandin E2 (PGE2) was significantly increased in macrophages in which inflammation was induced by LPS compared to cells in which inflammation was not induced, but was again decreased in cells treated with the sample. The concentration of prostaglandin E2 (PGE2) was decreased when 10 μg / ml of glycyl-L-leucine peptide or L-leucylglycine peptide was treated (Examples 2 and 4) compared to 1 μg / ml (Examples 1 and 3). In particular, it was confirmed that the amount of prostaglandin E2 (PGE2) secreted was decreased in cells treated with the composite composition (Examples 5 and 6) compared to cells treated with glycyl-L-leucine peptide or L-leucylglycine peptide alone (Examples 1 to 4).

[0119] The results of Experimental Example 2 suggest that a composition containing glycyl-L-leucine and / or L-leucylglycine may exert an effective anti-inflammatory effect by suppressing the secretion of prostaglandin E2 (PGE2), which induces inflammation, in inflammation-induced macrophages, and may also exert a fatigue-improving function.

[0120] Experimental Example 6: Antioxidant effect due to DPPH free radical scavenging activity DPPH (2,2-diphenyl-1-picrylhydrazyl) is a purple stable free radical that, when it reacts with an antioxidant, receives hydrogen and is reduced and decolorized. Using this principle, the antioxidant capacity of a sample can be evaluated by comparing the DPPH free radical scavenging activity of the sample. The higher the free radical scavenging activity (%), the higher the antioxidant capacity of the sample.

[0121] For the DPPH (1,1-diphenyl-2-picryl-hydrazyl) free radical scavenging activity, first, 4 mg of DPPH was dissolved in 100 ml of methanol to prepare a DPPH solution, and the composition of Experimental Example 1 was diluted with ethanol for each concentration. 1 ml of ethanol containing the composite composition was thoroughly mixed with 1 ml of DPPH solution, and then the mixture was left to stand at 37°C for 30 minutes, and the absorbance was measured at 516 nm. Ethanol was used as a control.

[0122] The antioxidant activity measured by the DPPH free radical scavenging activity method was calculated using the following formula.

[0123]

number

[0124] As a result, as shown in Figure 5, the DPPH free radical scavenging activity was increased when the samples were treated. The DPPH free radical scavenging activity was increased when glycyl-L-leucine peptide or L-leucylglycine peptide was treated at 10 μg / ml (Examples 2 and 4) compared to when glycyl-L-leucine peptide or L-leucylglycine peptide was treated at 1 μg / ml (Examples 1 and 3). In particular, it was confirmed that the DPPH free radical scavenging activity was increased in the samples of the composite composition (Examples 5 and 6) compared to the samples of glycyl-L-leucine peptide or L-leucylglycine peptide alone (Examples 1 to 4).

[0125] The results of Experimental Example 3 suggest that a composition containing glycyl-L-leucine and / or L-leucylglycine can be used as an effective antioxidant functional material by suppressing oxidative stress.

[0126] Experimental Example 7: Animal Experiments For the animal experiments, the inventors purchased male ICR mice (4 weeks old) from Daehan Biolink (Daejeon, Korea). The experimental animals were kept in a cage with 5-10 mice per cage in a breeding room maintained at a temperature of 22±1°C and humidity of 55±10%. Glycyl-L-leucine peptide or L-leucylglycine peptide, provided by Bachem AG (Bubendorf, Switzerland), was dissolved in distilled water and used. The control group was orally administered distilled water alone, and the glycyl-L-leucine peptide or L-leucylglycine peptide groups were orally administered once a day for 21 days using an oral administration sonde. The types and concentrations of the compositions administered to each group are shown in Table 2 below. Each group of mice was exercised once a week for 30 minutes for 3 weeks to induce fatigue. The exercise speed was 10 m / min for 10 minutes, 16 m / min for 10 minutes, and finally 21 m / min for 10 minutes. On the 21st day, the animals were exercised for 30 minutes at a speed of 10 m / min for the first 5 minutes, and then increased in speed by 3 m / min every 3 minutes. The animals were then sacrificed and used for analysis.

[0127] [Table 2]

[0128] Experimental Example 8: Preventing the accumulation of lactic acid in muscles improves fatigue In a situation where cells are sufficiently supplied with oxygen, the TCA cycle works smoothly, so that the blood lactate concentration is maintained in the range of 0.56 to 2 mmol / L and does not accumulate beyond that. However, in a situation where the metabolic cooperation between glycolysis and the TCA cycle is not performed at an equivalent ratio and glycolysis is relatively active, or in a situation where the oxygen supply is insufficient compared to the amount of oxygen required by cells, lactate is generated in the muscle. In addition, if the oxygen supply amount does not reach the amount of oxygen consumed by the muscle during high-intensity exercise, the lactate concentration in the muscle tissue increases, and the lactate generated at this time diffuses into the blood and is processed by the heart and liver, so if lactate accumulates due to exercise, it causes acidification in the body and induces fatigue. During exercise, phosphorylase activity involved in carbohydrate metabolism decreases, and as a result, the synthesis of glucose, which is a source of energy for exercise in an anaerobic state, is suppressed. Among various evaluations of fatigue improvement effects, the present inventors confirmed the suppression of accumulation of lactate and creatine kinase (CK) in the muscle, which indicate a fatigue state.

[0129] As a result, as shown in Figure 6, it was found that the concentration of lactic acid accumulated in the muscle was significantly reduced when glycyl-L-leucine peptide or L-leucylglycine peptide was orally administered (Examples 7 to 10) in a fatigue-induced mouse model compared to the control group (Comparative Example 2). In particular, it was confirmed that the concentration of lactic acid was reduced more when 10 mg / kg of glycyl-L-leucine peptide or L-leucylglycine peptide was ingested (Examples 8 and 10) than when 1 mg / kg / day (Examples 7 and 9) was ingested.

[0130] In other words, it was confirmed that in individuals with induced fatigue, the intake of glycyl-L-leucine peptide or L-leucylglycine peptide reduces the accumulation of lactic acid in the muscles and has the effect of improving fatigue.

[0131] Experimental Example 9: Preventing the accumulation of creatine kinase (CK) in muscles improves fatigue Creatine kinase (CK) is an enzyme found in large amounts in muscles, and when muscle cells are damaged, it is released into the blood to generate an energy source. Creatine kinase (CK) plays a role in synthesizing creatine phosphate, which is necessary for ATP resynthesis (the process of synthesizing ATP from ADP and phosphocreatine) in muscle cells during exercise when oxygen is absent.

[0132] To measure creatine kinase (CK) in muscle, fatigue was induced in mice, and muscle tissues were collected from each group of mice administered the composition and used for analysis, as in Experimental Example 4. Creatine kinase (CK) was measured using an analysis kit (Abcam, Cambridge, UK) according to the manufacturer's guidelines.

[0133] As a result, as shown in Figure 7, it was found that when glycyl-L-leucine peptide or L-leucylglycine peptide was orally administered (Examples 7 to 10) in a fatigue-induced mouse model, the concentration of creatine kinase (CK) accumulated in the muscle was significantly reduced compared to the control group (Comparative Example 2). In particular, it was confirmed that creatine kinase (CK) was reduced more when 10 mg / kg of glycyl-L-leucine peptide or L-leucylglycine peptide was ingested (Examples 8 and 10) than when 1 mg / kg / day of glycyl-L-leucine peptide or L-leucylglycine peptide was ingested (Examples 7 and 9).

[0134] In other words, it was confirmed that in individuals with induced fatigue, the intake of glycyl-L-leucine peptide or L-leucylglycine peptide reduces the accumulation of creatine kinase (CK) in the muscles and has the effect of improving fatigue.

[0135] Experimental Example 10: Inflammation improvement effect by reducing cytokines in the blood Representative cytokines include interleukin (IL), interferon (IFN), chemokine, tumor necrosis factor (TNF), and transforming growth factor (TGF), among which pro-inflammatory cytokines include interleukin-1β, -2, -6 (IL-1β, -2, -6), IFN-γ, and TNF-α, while anti-inflammatory cytokines include IL-4, IL-10, IL-11, IL-13, and TGF-β. The inventors have confirmed the inhibition of secretion of tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6), which are inflammation-inducing proteins, in the evaluation of inflammation-improving effects.

[0136] First, to measure blood tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6), fatigue was induced as in Experimental Example 4, and blood was collected from mice in each composition-administered group. The collected blood was left at room temperature for 30 minutes, and then centrifuged at 6000 rpm for 20 minutes to separate serum. The separated serum was filtered through a 0.45 μm syringe filter and used for analysis. Tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) were measured using commercially available TNF-α and IL-6 ELISA kits (Thermo Scientific Co., Waltham, MA, USA) according to the manufacturer's guidelines.

[0137] As a result, as shown in Figure 8, in a fatigue-induced mouse model, when glycyl-L-leucine peptide or L-leucylglycine peptide was orally administered (Examples 7 to 10), the blood tumor necrosis factor-α (TNF-α) concentration was significantly reduced compared to the control group (Comparative Example 2). In particular, tumor necrosis factor-α (TNF-α) was reduced more when glycyl-L-leucine peptide or L-leucylglycine peptide was administered at 10 mg / kg / day (Examples 8 and 10) than when glycyl-L-leucine peptide or L-leucylglycine peptide was administered at 1 mg / kg / day (Examples 7 and 9).

[0138] In addition, as shown in Figure 9, when glycyl-L-leucine peptide or L-leucylglycine peptide was orally administered to a fatigue-induced mouse model, the blood interleukin-6 (IL-6) concentration was significantly reduced compared to the control group (Comparative Example 2). In particular, interleukin-6 (IL-6) was reduced more when 10 mg / kg of glycyl-L-leucine peptide or L-leucylglycine peptide was administered (Examples 8 and 10) than when 1 mg / kg of glycyl-L-leucine peptide or L-leucylglycine peptide was administered (Examples 7 and 9). In other words, it was confirmed that in individuals with induced fatigue, the intake of glycyl-L-leucine peptide or L-leucylglycine peptide reduced the concentration of inflammatory cytokines in the blood and had an inflammation-improving effect.

[0139] Experimental Example 11: Antioxidant effect by increasing blood superoxide dismutase (SOD) Superoxide dismutase (SOD) is an antioxidant enzyme that catalyzes the reaction of converting superoxide into oxygen and hydrogen peroxide (H2O2) and plays a role in removing free radicals from the blood. An increase in blood SOD reduces fatigue in muscles. To evaluate the antioxidant effect, the inventors confirmed an increase in superoxide dismutase (SOD), which is responsible for the body's antioxidant function.

[0140] To measure the level of superoxide dismutase (SOD) in blood, fatigue was induced in mice of each group administered with the composition in the same manner as in Experimental Example 4, and serum obtained from the mice was used for analysis. Superoxide dismutase (SOD) was measured using an analysis kit (Abcam, Cambridge, UK) according to the manufacturer's instructions.

[0141] As a result, as shown in Figure 10, when glycyl-L-leucine peptide or L-leucylglycine peptide was orally administered to a fatigue-induced mouse model (Examples 7 to 10), the concentration of superoxide dismutase (SOD) in the blood was significantly increased compared to the control group (Comparative Example 2). In particular, superoxide dismutase (SOD) increased more when 10 mg / kg of glycyl-L-leucine peptide or L-leucylglycine peptide was ingested (Examples 8 and 10) than when 1 mg / kg of glycyl-L-leucine peptide or L-leucylglycine peptide was ingested (Examples 7 and 8).

[0142] In other words, it was confirmed that in individuals with induced fatigue, the ingestion of glycyl-L-leucine peptide or L-leucylglycine peptide increased the concentration of superoxide dismutase (SOD) in the blood, thereby exerting an antioxidant effect.

[0143] Experimental Example 12: Estrogen receptor activity evaluation "Menopausal symptoms" refer to the many symptoms that occur in women around the time of menopause due to a decrease in estrogen secretion caused by ovarian aging and other factors.

[0144] To confirm the activity of the two dipeptides of formulas 1 and 2 against the estrogen receptor, an estrogen response element (ERE) reporter assay was performed. 293T cells were cultured in a 24-well plate using DMEM medium supplemented with 10% FBS for 24 hours, and then the medium was replaced with 500 μl of 5% charcoal-treated FBS (phenol red-free) per well and transfection was performed. 0.1 μg of ERa, 0.1 μg of ERE, and 10 ng of pRL-Tk DNA per well were transfected using Lipofectamine® reagent (Thermo Fisher Scientific). Four hours after transfection, the composition of Experimental Example 1 was treated, and 1 ppb of 17β-estradiol (E2) (DMSO 0.5 μl) was treated as a positive control, and the same amount of DMSO (0.5 μl) was treated as a negative control (control, ctrl). 24 hours after sample treatment, the luminescence level was measured using the Dual-Luciferase® Reporter Assay System (Promega), and the negative control group was set as the standard (1.0). The firefly luciferase value was corrected to the Renilla luciferase value.

[0145] As a result, as shown in Figure 11, the activity of the estrogen receptor was increased when the sample was treated. The activity of the estrogen receptor was increased when 10 μg / ml of glycyl-L-leucine peptide or L-leucylglycine peptide was treated (Examples 2 and 4) compared to when 1 μg / ml of the peptide was treated (Examples 1 and 3). In particular, it was confirmed that the activity of the estrogen receptor was increased in the samples of the composite composition (Examples 5 and 6) compared to the samples of the single glycyl-L-leucine peptide or L-leucylglycine peptide (Examples 1 to 4).

[0146] Experimental Example 13: Inhibitory effect on osteoclast differentiation During menopause, the ovaries stop secreting female hormones, which play an important role in bone metabolism, causing changes in bone turnover. Although osteoblasts regenerate the areas destroyed by osteoclasts, this is not complete, and bone loss occurs. Bone loss occurs gradually from the 30s onwards, and in men, bone density gradually decreases, resulting in a loss of about 20-30% of maximum bone mass. Meanwhile, in women, bone density decreases rapidly in their 50s, i.e., for about 10 years after menopause, and then decreases more slowly, causing osteoporosis to progress faster and more severely than in men.

[0147] In vitro, osteoclasts can be generated from macrophage precursor cells by the specific cytokine, RANKL (RANK ligand), and when stained with TRAP (tartrate-resistant acid phosphatase), the control group not treated with RANKL shows that RAW264.7 cells proliferate while maintaining their spherical shape, and show a negative reaction to TRAP staining, taking on a pale brown or ochre color.On the other hand, in the cells treated with RANKL, TRAP(+) multinucleated cells stained dark brown or reddish brown are observed.

[0148] To confirm the effect of the two dipeptides of formulas 1 and 2 on osteoclast differentiation induced by NF-κB activator ligand (RANKL), RAW264.7 cells cultured in Experimental Example 5 were used in the experiment. RAW264 cells were cultured at 2.5×10 6 The cells were dispensed at a cell concentration of 100 μl per well into a 96-well microplate at 100 cells / ml, and then cultured for 5 hours until the cells attached to the wells. The medium was removed, and a culture solution prepared by mixing 50 ng / mL of RANKL, an osteoclast differentiation factor, and the composition of Experimental Example 1 in α-MEM medium supplemented with 10% FBS was dispensed and cultured for 7 days. To evaluate the effect on osteoclast differentiation, the cultured cells were stained with TRAP, and the number of cells stained red was counted, assuming that they were osteoclasts.

[0149] As a result, as shown in Figure 12, osteoclast differentiation was suppressed when the samples were treated. The number of osteoclasts was reduced when 10 μg / ml of glycyl-L-leucine peptide or L-leucylglycine peptide was treated (Examples 2 and 4) compared to when 1 μg / ml of glycyl-L-leucine peptide or L-leucylglycine peptide was treated (Examples 1 and 3). In particular, it was confirmed that osteoclast differentiation was suppressed in the samples of the composite composition (Examples 5 and 6) compared to the samples of glycyl-L-leucine peptide or L-leucylglycine peptide alone (Examples 1 to 4).

[0150] The results of Experimental Examples 12 and 13 suggest that a composition containing glycyl-L-leucine and / or L-leucylglycine can be effectively used for treating female menopausal symptoms by increasing estrogen receptor activity and inhibiting osteoclast differentiation, which is a factor inducing osteoporosis after menopause.

[0151] From the above description, a person skilled in the art to which the present invention pertains can understand that the present invention can be implemented in other specific forms without changing the technical idea or essential features of the present invention. It should be understood that the above-described embodiments are illustrative and not limiting. Furthermore, it goes without saying that the scope of the present invention includes any modifications or variations derived from the above detailed description, the meaning and scope of the claims described below, and their equivalent concepts.

Claims

1. An agent for treating, preventing or improving menopausal symptoms in women, comprising a compound represented by the following chemical formula 1 and / or chemical formula 2 or a salt thereof in an amount effective for treating, preventing or improving menopausal symptoms in women. 【Chemistry 1】

2. 2. The agent for treating, preventing or improving menopausal symptoms in women according to claim 1, wherein the menopausal symptoms in women include at least one selected from the group consisting of facial flushing, sweating, nervousness, depression, dizziness, fatigue, joint pain, muscle pain, headache, chest palpitations, feeling of formication, sweating during sleep, sleep disorders, dry skin, vaginal dryness, vaginal atrophy, lower urethral atrophy, pain during intercourse, vaginitis, cystitis, painful urination, urgent urination, concentration disorder, memory disorder, anxiety, irritability, memory decline, dry skin, joint pain, osteoporosis, heart disease, high blood pressure and stroke.

3. A food composition comprising the agent for treating, preventing or ameliorating menopausal symptoms in women according to claim 1 or 2.

4. A cosmetic composition comprising the agent for treating, preventing or ameliorating menopausal symptoms in women according to claim 1 or 2.

5. A pharmaceutical composition comprising the agent for treating, preventing or ameliorating menopausal symptoms in women according to claim 1 or 2.

6. A quasi-pharmaceutical composition comprising the agent for treating, preventing or ameliorating menopausal symptoms in women according to claim 1 or 2.

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