KR102652809B1 - Composition for improving skin conditions - Google Patents

Abstract

The present invention relates to a composition for improving skin condition. 11-keto-beta-boswellic acid according to the present invention increases the expression of antibacterial peptides in keratinocytes, improves skin immunity and improves skin defense through antibacterial effects, increases skin moisture content, and increases the concentration of hyaluronic acid in fibroblasts. It promotes the production of hyaluronic acid, increases the expression of hyaluronic acid synthase, and improves skin elasticity, improving skin moisturization and skin elasticity. At the same time, it is non-cytotoxic and has no side effects on the human body, so it can be used in pharmaceuticals, cosmetics, food, or external skin applications. It can be used in formulation manufacturing.

Description

Composition for improving skin conditions}

The present invention relates to a composition for improving skin condition that improves skin defense, improves skin moisturization, and improves skin elasticity.

Recently, extrinsic skin diseases are increasing significantly due to industrial development and changes in lifestyle patterns. In particular, as the living environment of urban residents changes recently, the opportunity to be exposed to exogenous pathogens, such as mites living in carpets at home, bacteria and fungi that reproduce depending on humidity and temperature, and viruses in public places, is increasing. To combat these exogenous pathogens, immunity in the skin, the outermost organ of the human body, is becoming important. Among the skin's defense mechanisms, antibacterial peptides are substances responsible for in vivo defense techniques other than physical defense. There are two main types of antibacterial peptides: cathelicidin and defensin. Cathelicidin exhibits a wide range of antibacterial activities and has various immunomodulatory functions. Among them, LL-37 is the only known cathelicidin expressed in humans, and its production is determined by factors such as the presence of bacteria, secretion of host cytokines, amount of oxygen available in the body, and vitamin D. LL-37 plays a role in causing a biological immune response to external tissue wounds or infections. It has direct antibacterial activity against bacteria, fungi, and viruses, as well as chemotaxis against neutrophils (eosinophils), monocytes, and T cells, and endothelial cells. induces the proliferation of In particular, LL-37 present in the skin plays an antigen-suppressing function by acting as an immediate defense when foreign antigens penetrate.

Defensin is one of the most studied antibacterial peptides, and is largely divided into α-defensin and β-defensin depending on its structural characteristics. Among them, β-defensin is a peptide substance expressed in the mucosal epithelium of the skin, lungs, organs, kidneys, genitals, etc. To date, there are six types of human-derived β-defensins, namely human β-defensin-1 (hBD1). , human β-defensin-2 (hBD2), human β-defensin-3 (hBD3), human β-defensin-4 (hBD4), human β-defensin-5 (hBD5) and human β-defensin. -6(hBD6) has been isolated and identified. In particular, while hBD1 is constantly expressed in epidermal cells, hBD2 is expressed increased at sites of infection or physical damage, and is known to play an important role in regulating systemic immune and inflammatory responses. In addition, hBD3 has been reported to be highly expressed in skin lesions of patients with psoriasis disease and is inducibly expressed by cytokines and bacterial products. hBD4 shows a more restricted distribution compared to hBD1, hBD2, or hBD3, and its expression can be upregulated by bacterial infection, but not by inflammatory factors that upregulate hBD2 and hBD3. hBD5 and hBD6 are the most recently identified family members located primarily in the epithelium. In addition, β-defensin has recently been shown to be involved in not only local infection defense but also adaptive immunity by causing chemotactic migration of cells such as dendritic cells, T lymphocytes, and monocytes. It is known.

Additionally, sensitive skin is prone to skin immune diseases such as atopy and psoriasis when exposed to harmful environments. Conventionally, the improvement of skin immune diseases such as atopy and psoriasis has been mainly solved by maintaining moisturization using ceramide, but there is a problem with moisturizing that the moisturizing power decreases over time and the skin condition returns to its original state. A method of producing antibacterial peptides externally and injecting them into the skin can be considered, but since direct penetration into the skin is difficult and there is a risk of autoimmune reactions, the method of promoting the production of antibacterial peptides by skin cells themselves is recommended. It could be a way to increase immunity from the outside and have antibacterial properties.

In addition, the epidermis, located in the outermost layer of the skin, performs a protective function to defend against various external physical, chemical and mechanical stimuli and prevent excessive dissipation of body moisture through the skin. This protective function is possible through the normal formation and maintenance of the stratum corneum, which is composed of keratinocytes. Keratinocytes are cells that are formed through gradual changes in form and function as basal cells, which continuously proliferate in the lowermost layer of the epidermis (stratum basale), move to the stratum corneum. After a certain period of time, old keratinocytes are shed from the skin, and new keratinocytes from the lowest layer of the epidermis take over their functions, repeating the process of epidermis differentiation or keratinization. During this keratinization process, keratinocytes produce Natural Moisturizing Factor (NMF) and intercellular lipids such as ceramide, cholesterol, and fatty acids, and the stratum corneum acts as a barrier to the outside, forming a skin barrier. It has the function of

In this regard, dry skin, which is considered one of the major diseases in modern society, is one of the symptoms caused by abnormal skin barrier function. Recently, it has been increasing due to various factors such as environmental pollution, increase in dry environments such as apartments and high-rise buildings, increase in social stress, excessive bathing culture unique to Korea, and skin aging. Cases are also continuously increasing.

However, the use of the above-mentioned moisturizers is mostly only a temporary relief of symptoms rather than a fundamental treatment, and does not show sufficient effectiveness in treating dry skin, including atopic dermatitis, and skin barrier dysfunction. Accordingly, there is an urgent need to develop a material that fundamentally regenerates the damaged skin barrier.

Hyaluronic acid is a type of glycosaminoglycans and is a chain-shaped polymer polysaccharide in which glucuronic acid and N-acetylglucosamine residues are repeatedly linked. It has the property of forming a gel by combining with a large amount of water, so it has high viscosity and elasticity. Hyaluronic acid is a major component of the extracellular matrix and has been reported to be involved in water retention, maintenance of intercellular space, storage and diffusion of cell growth factors and nutrients, as well as cell division, differentiation, and migration. there is.

It has been reported that more than 50% of the hyaluronic acid present in the mammalian body is distributed in the skin, especially in the intercellular spaces of the epidermis and the connective tissue of the dermis. This hyaluronic acid is known to be mainly synthesized by keratinocytes and fibroblasts. . The amount of hyaluronic acid in human skin has been reported to decrease with aging, and the decrease in hyaluronic acid in the skin is believed to be one of the direct causes of decreased skin elasticity and moisture content due to aging ( Biochem Biophys Acta 279, 265-275; Carbohydr Res 159, 127-136; Int J Dermatol 33, 119-122). In addition, hyaluronic acid is known to be involved in maintaining the structure of the stratum corneum and skin barrier function ( J Cosmet Dermatol . 2007 Jun, 6(2), 75-82).

However, hyaluronic acid, which has the above effects, has a large molecular weight and is not easily absorbed into the skin. In addition, a method of injecting hyaluronic acid into the skin using an injection is currently being performed, but this method not only causes great irritation, but a method of increasing hyaluronic acid synthesis within skin cells is more effective. Therefore, research is being actively conducted on methods to increase the production of hyaluronic acid in the human body, but no notable research results are yet known.

Therefore, the development of ingredients that are safe when applied to the human body, have stable active ingredients, and, above all, are superior to existing substances in improving skin immunity for atopic dermatitis and psoriasis, enhancing skin defense through enhancing antibacterial properties, improving skin moisturization, and enhancing skin elasticity. It is urgently needed.

Ann Dermatol. Vol. 24(2), pp. 126-135 (2012.04.26) Br J Dermatol. Vol. 169(3), pp. 587-593 (2013. 09) Biochem Biophys Acta, vol. 279, pp. 265-275 (1972) Carbohydr Res, vol. 159, pp. 127-136 (1987) Int J Dermatol, vol. 33, pp. 119-122 (1994) J Cosmet Dermatol., vol. 6(2), pp. 75-82 (2007. 06)

The purpose of the present invention is to provide a use of a compound represented by the following formula (1), which has fewer side effects such as skin irritation when applied to the human body and can improve skin conditions such as improving skin defense, improving skin moisturization, and improving skin elasticity:

[Formula 1]

As a means to solve the above problems, the present invention provides a pharmaceutical, cosmetic, food, or external skin preparation formulation for improving skin defense, improving skin moisturization, and improving skin elasticity, comprising a compound represented by the following Chemical Formula 1 as an active ingredient: Provided is a composition:

[Formula 1]

The present invention also provides a method for enhancing skin defense, improving skin moisturization, and improving skin elasticity, comprising the step of applying the compound of Formula 1 or a cosmetically acceptable salt thereof to the skin of an individual.

11-keto-beta-boswellic acid according to the present invention increases the expression of antibacterial peptides in keratinocytes, improves skin immunity and improves skin defense through antibacterial effects, increases skin moisture content, and increases the concentration of hyaluronic acid in fibroblasts. It promotes the production of hyaluronic acid, increases the expression of hyaluronic acid synthase, and improves skin elasticity, improving skin moisturization and skin elasticity. At the same time, it is non-cytotoxic and has no side effects on the human body, so it can be used in pharmaceuticals, cosmetics, food, or external skin applications. It can be used in formulation manufacturing.

Hereinafter, the configuration of the present invention will be described in detail.

In order for ingredients to improve skin immunity and antibacterial power to improve skin defense, improve skin moisturization, and improve skin elasticity to have excellent effects when applied to actual skin, they must be highly active at low concentrations to improve skin defense, improve skin moisturization, and promote skin elasticity. It has excellent ability to penetrate and be absorbed through the skin, has low volatility so that it can remain for a sufficient period of time to improve skin defense, improve skin moisturization, and improve skin elasticity, and the active ingredient remains stable in the composition or on the skin, It is desirable that it is easy to formulate into pharmaceuticals, cosmetics, food, external skin preparations, etc., and is also safe for the skin. However, among known components, components that satisfy all of the above characteristics are rare. For example, some ingredients that enhance skin defense, improve skin moisturization, and improve skin elasticity are excellent in enhancing skin defense, improving skin moisturizing, and improving skin elasticity even at low concentrations in in vitro experiments, but their ability to penetrate and absorb through the skin is low, so they are not effective in improving skin elasticity. It is difficult to apply to the skin. Other active ingredients have low hydrophilicity, making them difficult to formulate into medicines, cosmetics, or foods. In addition, some ingredients that enhance skin defense, improve skin moisturization, and improve skin elasticity may lose their effectiveness before application to the skin because the active ingredients are decomposed or transformed into other compounds when exposed to heat, light, or oxygen.

As can be seen in the examples below, 11-keto-beta-boswellic acid has no cytotoxicity and therefore has no side effects on the human body, maintains stability within the composition, and increases the expression of antibacterial peptides in keratinocytes, thereby improving skin immunity. It improves skin defense through improvement and antibacterial activity, increases skin moisture content, promotes the production of hyaluronic acid in fibroblasts, increases the expression of hyaluronic acid synthase, and at the same time improves skin elasticity, compared to conventional active ingredients. Since it has excellent skin moisturizing and skin elasticity enhancement effects, it can be used as an active ingredient in pharmaceuticals, cosmetics, foods, and external skin preparations to improve skin defense, improve skin moisturization, and enhance skin elasticity.

Accordingly, the present invention provides a composition for improving skin defense, improving skin moisturization, and improving skin elasticity, comprising a compound represented by the following formula (1) as an active ingredient.

[Formula 1]

The compound name of the compound of Formula 1 is also called 11-keto-beta-boswellic acid.

The compound of Formula 1 can be synthesized and used, or a commercially available compound can be used.

The compositions of the present invention can be used for preparing pharmaceutical formulations.

Accordingly, the composition of the present invention may include a pharmaceutically acceptable salt of the compound of Formula 1 above.

Pharmaceutically acceptable salts of the compound of Formula 1 may be acid addition salts formed using organic acids or inorganic acids, and the organic acids include, for example, formic acid, acetic acid, propionic acid, lactic acid, butyric acid, isobutyric acid, and trifluoroacetic acid. , malic acid, maleic acid, malonic acid, fumaric acid, succinic acid, succinic acid monoamide, glutamic acid, tartaric acid, oxalic acid, citric acid, glycolic acid, glucuronic acid, ascorbic acid, benzoic acid, phthalic acid, salicylic acid, anthranilic acid, dichloroacetic acid, aminooxy acetic acid. , benzenesulfonic acid, p-toluenesulfonic acid, and methanesulfonic acid-based salts, and inorganic acids include, for example, hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, nitric acid, carbonic acid, and boric acid-based salts. Preferably it may be in the form of hydrochloride or acetate, and more preferably in the form of hydrochloride.

The above-mentioned acid addition salts can be prepared by a) mixing the compound of formula 1 and the acid directly, b) dissolving and mixing one of them in a solvent or hydrous solvent, or c) mixing the compound of formula 1 in a solvent or water. It is prepared by a general salt preparation method of placing an acid in a solvent and mixing them.

Apart from the above, additional salt forms available include gaba salt, gabapentin salt, pregabalin salt, nicotinate salt, adipate salt, hemimalonate, cysteine salt, acetylcysteine salt, methionine salt, arginine salt, lysine salt, ornithine salt, Aspartate, etc.

The composition may further contain one or more active ingredients that exhibit the same or similar functions. For example, it may contain ingredients known to improve skin defense, improve skin moisturization, and improve skin elasticity. If additional skin defense enhancement, skin moisturizing, and skin elasticity enhancement ingredients are included, the skin defense enhancement, skin moisturization, and skin elasticity enhancement effects of the composition of the present invention can be further enhanced. When adding the above ingredients, skin safety due to combined use, ease of formulation, and stability of the active ingredients can be taken into consideration. In one embodiment of the present invention, the composition contains substances that inhibit tyrosinase enzyme activity such as kojic acid and arbutin, hydroquinone, and vitamin-C (L-Ascorbic) known in the art. acid) and their derivatives, or various plant extracts, etc. may be included alone or in combination of two or more. Additional ingredients may be included in an amount of 0.0001% to 10% by weight based on the total weight of the composition, and the content range may be adjusted according to requirements such as skin safety and ease of formulation of the compound of Formula 1. .

Additionally, the composition of the present invention may further include a pharmaceutically acceptable carrier.

Pharmaceutically acceptable carriers may contain various ingredients such as buffers, sterile water for injection, normal saline or phosphate buffered saline, sucrose, histidine, salts and polysorbates.

The composition of the present invention can be administered orally or parenterally, and can be administered in the form of a general pharmaceutical preparation, for example, various oral and parenteral formulations during clinical administration. When formulated, commonly used fillers and extenders are used. It can be prepared using diluents or excipients such as binders, wetting agents, disintegrants, and surfactants.

Solid preparations for oral administration include tablets, pills, powders, granules, capsules, etc. These solid preparations include at least one excipient in the pharmaceutical composition of the present invention, such as starch, calcium carbonate, It can be prepared by mixing sucrose, lactose, gelatin, etc.

In addition to simple excipients, lubricants such as magnesium styrate talc are also used. Liquid preparations for oral use include suspensions, oral solutions, emulsions, syrups, etc. In addition to the commonly used simple diluents such as water and liquid paraffin, various excipients such as wetting agents, sweeteners, fragrances, and preservatives may be included. .

Preparations for parenteral administration include sterilized aqueous solutions, non-aqueous solutions, suspensions, emulsions, freeze-dried preparations, and suppositories. Non-aqueous solvents and suspensions may include propylene glycol, polyethylene glycol, vegetable oil such as olive oil, and injectable ester such as ethyl oleate. As a base for suppositories, witepsol, macrogol, tween 61, cacao, laurin, glycerogeratin, etc. can be used.

In this specification, 'skin defense' is the ability to protect the skin from external stimuli and toxins, and the stimuli are a broad concept that includes all stimuli harmful to the skin, such as bacteria, viruses, ultraviolet rays, dryness, and wounds. More specifically, it includes enhancing skin defense by improving skin immunity and enhancing antibacterial power.

In this specification, 'skin immunity' refers to antibacterial peptides that play a role in causing a biological immune response against external pathogens such as external tissue wounds or infections, and have direct antibacterial activity against bacteria, fungi, viruses, etc., as well as neutrophils (eosinophils). ), exhibits chemotaxis to monocytes and T cells, induces the proliferation of endothelial cells, and in particular, antibacterial peptides present in the skin serve as an immediate defense upon penetration of foreign antigens, thereby serving an antigen-suppressing function.

In this specification, the term 'antibacterial' means that antibacterial peptides exist in an inactivated form in specific neutrophil granules against various microorganisms, including bacteria, fungi, and viruses, and are then cleaved and secreted out of the cell in an activated form. It refers to the function of destroying the cell membrane of a microorganism by binding the cationic portion of the antibacterial peptide to the anionic phospholipid of the microbial cell membrane.

In this specification, the term 'moisturizing effect' refers to suppressing or inhibiting the decrease in skin moisture or increasing the moisture content of the skin to smooth the skin surface and provide gloss. Alternatively, it means maintaining body homeostasis by appropriately controlling moisture loss (water evaporation).

In this specification, 'skin elasticity' is expressed by elastic fibers composed of elastin present in the dermal layer. These elastic fibers have a very low elastic modulus like rubber, so they are easily deformed even by a small force, and the force When removed, it easily returns to its original shape. When hyaluronic acid, which connects elastin and collagen, decreases, skin elasticity decreases. Therefore, the 'elasticity enhancement effect' refers to an increase in the elasticity of the skin, suppressing or inhibiting the loss of skin elasticity, or alleviating already reduced elasticity.

In this specification, 'improvement' refers to all actions in which the various skin conditions mentioned above are improved or beneficially changed due to the administration of a composition containing an active ingredient.

In this specification, 'skin' refers to the tissue that covers the body surface of an animal, and is the broadest concept that includes not only the tissue that covers the body surface such as the face or body, but also the scalp and hair.

In the present invention, the 'effective amount' refers to increasing the expression of antibacterial peptides in keratinocytes, improving skin immunity and enhancing skin defense through antibacterial effects, increasing skin moisture, and increasing the production of hyaluronic acid in fibroblasts. It refers to the amount of a compound that can improve skin condition by promoting the expression of hyaluronic acid synthase and improving skin elasticity, thereby improving skin moisturization and improving skin elasticity. When the composition of the present invention contains an effective amount of the compound of Formula 1, it can provide desirable effects of improving skin defense, improving skin moisturization, and improving skin elasticity. The effective amount of the compound of Formula 1 included in the composition of the present invention will vary depending on the form in which the composition is commercialized, the method in which the compound is applied to the skin, and the time it remains on the skin. For example, when the composition is commercialized as a pharmaceutical formulation, it may contain the compound of Formula 1 at a higher concentration than when it is commercialized as a cosmetic product that is routinely applied to the skin. Therefore, the daily dosage is 0.1 to 100 mg/kg, preferably 30 to 80 mg/kg, more preferably 50 to 60 mg/kg, based on the amount of the compound of Formula 1, and 1 to 60 mg/kg per day. Can be administered 6 times.

The composition of the present invention can be used alone or in combination with surgery, radiation therapy, hormone therapy, chemical therapy, and methods using biological response modifiers.

In addition, the composition for improving skin defense, improving skin moisturization, and improving skin elasticity of the present invention can be used to prepare a skin external preparation formulation.

When the compound of Formula 1 is used as an external skin agent, fatty substances, organic solvents, solubilizers, thickeners and gelling agents, softeners, antioxidants, suspending agents, stabilizers, foaming agents, fragrances, and surfactants are added. , water, ionic or non-ionic emulsifiers, fillers, sequestering agents and chelating agents, preservatives, vitamins, blocking agents, humectants, essential oils, dyes, pigments, hydrophilic or lipophilic actives, lipid vesicles or in external preparations for the skin. It may contain adjuvants commonly used in the field of dermatology, such as any other ingredients used. Additionally, the ingredients may be introduced in amounts commonly used in the field of dermatology.

When the compound of Formula 1 is provided in a formulation for external skin application, it may have a formulation such as an ointment, patch, gel, cream, or spray, but is not limited thereto.

Additionally, the composition for improving skin defense, improving skin moisturization, and improving skin elasticity of the present invention can be used to manufacture cosmetic formulations.

The cosmetic formulation may be in the form of a general emulsified formulation or solubilized formulation. For example, lotion such as flexible lotion or nourishing lotion, emulsion such as facial lotion, body lotion, cream, essence, cosmetic ointment, spray, gel, pack, sunscreen, makeup base, liquid such as nutritional cream, moisture cream, eye cream, etc. It may have a formulation such as foundation type, solid type or spray type, powder, makeup remover such as cleansing cream, cleansing lotion, cleansing oil, cleansing foam, soap, body wash, etc.

In addition to the compound of Formula 1, the cosmetics include fatty substances, organic solvents, solubilizers, thickeners and gelling agents, softeners, antioxidants, suspending agents, stabilizers, foaming agents, fragrances, surfactants, and water. , ionic or non-ionic emulsifiers, fillers, sequestering and chelating agents, preservatives, vitamins, blocking agents, wetting agents, essential oils, dyes, pigments, hydrophilic or lipophilic active agents, lipid vesicles or commonly used in cosmetics. It may contain auxiliaries commonly used in the field of cosmetology, such as any other ingredients.

The cosmetic formulation may contain a relatively high concentration of the compound of Formula 1 in the case of wash-off type cosmetics such as makeup removers and detergents in which the active ingredient stays on the skin for a short period of time. On the other hand, in the case of leave-on type cosmetics such as lotions, emulsions, creams, and essences, where the active ingredients stay on the skin for a long period of time, the compound of Formula 1 is used at a lower concentration than wash-off type cosmetics. It would be okay to include it. Although not limited thereto, in one embodiment of the present invention, the composition may include the compound of Formula 1 in an amount of 0.0001% by weight to 10% by weight (preferably 0.0001% by weight to 1% by weight) based on the total weight of the composition. You can. If the composition of the present invention contains less than 0.0001% by weight of the compound of Formula 1, sufficient skin defense enhancement, skin moisturizing, and skin elasticity enhancement effects cannot be expected, and if it contains more than 10% by weight, allergies, etc. This is to prevent unwanted reactions or skin safety issues.

Additionally, the composition for improving skin defense, improving skin moisturization, and improving skin elasticity of the present invention can be used for manufacturing food formulations.

The food formulation refers to a food manufactured by adding the compound of Chemical Formula 1 to food materials such as beverages, teas, spices, gum, and confectionery, or by encapsulating, powdering, or suspending it.

Since the food formulation can be consumed on a daily basis, it is very useful as it can be expected to enhance skin defense, improve skin moisturization, and improve skin elasticity.

When using the compound of Formula 1 as a food additive, the compound of Formula 1 can be added as is or used together with other foods or food ingredients, and can be used appropriately according to conventional methods. The mixing amount of the active ingredient can be appropriately determined depending on the purpose of use (prevention, health, or therapeutic treatment). Generally, 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 materials. However, in the case of long-term intake for the purpose of health and hygiene or health control, the amount 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 are no special restrictions on the types of foods above. Examples of foods to which the above substances can be added include meat, sausages, bread, chocolate, candies, snacks, confectionery, pizza, ramen, other noodles, gum, dairy products including ice cream, various soups, beverages, tea, drinks, etc. These include alcoholic beverages and vitamin complexes, and include all health foods in the conventional sense.

When the food formulation is a beverage, it may contain various flavoring agents or natural carbohydrates as additional ingredients, like regular beverages. The above-mentioned natural carbohydrates include monosaccharides such as glucose and fructose, disaccharides such as maltose and sucrose, polysaccharides such as dextrin and cyclodextrin, and sugar alcohols such as xylitol, sorbitol, and erythritol. As a sweetener, natural sweeteners such as thaumatin and stevia extract or synthetic sweeteners such as saccharin and aspartame can be used. The proportion of the natural carbohydrate is generally about 0.01 to 0.04 g, preferably about 0.02 to 0.03 g, per 100 mL of the composition of the present invention.

In addition to the above, food formulations include various nutrients, vitamins, electrolytes, flavors, colorants, pectic acid and its salts, alginic acid and its salts, organic acids, protective colloidal thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohol, carbonated drinks. It may contain carbonating agents used in. Additionally, the food formulation may contain pulp for the production of natural fruit juices, fruit juice drinks and vegetable drinks. These ingredients can be used independently or in combination. The ratio of these additives is not very important, but is generally selected in the range of 0.01 to 0.1 parts by weight per 100 parts by weight of the composition of the present invention.

The present invention also relates to a method for enhancing skin defense, improving skin moisturization, and improving skin elasticity, comprising the step of applying the compound of Formula 1 or a cosmetically acceptable salt thereof to the skin of an individual.

The subject includes, but is not limited to, mammals such as rats, pigs, and humans.

Hereinafter, the present invention will be described in detail by examples. However, the following examples only illustrate the present invention, and the content of the present invention is not limited to the following examples.

<Reference Example 1> 11-keto-beta-boswellic acid information

[Formula 1]

CAS No.: 17019-92-0

Where to buy: Sigma Aldrich

<Example 1> Cytotoxicity evaluation

The compound of Formula 1 was added to the culture medium of normal human keratinocytes and cytotoxicity was evaluated at the cellular level.

For this purpose, the compound of Formula 1 was treated at different concentrations and cell viability was confirmed using a spectrophotometer. Human-derived keratinocytes were placed in a 24-well test plate at a rate of 1 × 10 ⁵ cells, allowed to attach, and cultured for 24 hours. When the cells reached 90% or more, they were treated with the compound of Formula 1 at concentrations of 2 ㎍/mL and 5 ㎍/mL, respectively. did. After culturing for 24 hours, the medium was removed, 200 μl of medium and 20 μl of CCK8 solution were added to each well, and cultured in an incubator at 37°C and 5% CO ₂ for 20 minutes, and then the absorbance was measured at 450 nm. The effect of the compound of Formula 1 on cell survival was evaluated based on the absorbance of untreated wells, and the experiment was repeated three times.

sample absorbance Survival rate (%) Control group (no additives) 0.595 ± 0.055 - Compound of Formula 1
(2 ㎍/mL) 0.599 ± 0.119 100% Compound of Formula 1
(5 ㎍/mL) 0.589 ± 0.043 98%

As shown in Table 1, it was confirmed that the compound of Formula 1 did not cause cytotoxicity at the experimental concentration. This means that the compound of Formula 1 has a low possibility of causing skin irritation and is a safe substance. In addition, it was found that the compound of Formula 1 was harmless to keratinocytes even at a concentration of 5 μg/mL.

<Example 2> Antibacterial activity evaluation

To evaluate the antibacterial properties of the compound of Formula 1, it was added to the culture medium of normal human keratinocytes and its effect on the expression of antibacterial peptides, LL37 and hBD2 mRNA, was measured at the cellular level.

For this purpose, 5 × 10 ⁴ cells of human-derived keratinocytes were placed and attached to a 24-well test plate, cultured for 1 day, and when the cell reached 80% or more, the compound of Formula 1 was added at 2 ㎍/mL or 5 ㎍/mL. Processed. 24 hours after treatment, cells were recovered, total RNA was extracted, the expression levels of antibacterial peptides LL37 and hBD2 mRNA were measured, and the experiment was repeated three times.

sample LL37 relative expression level (fold) hBD2 relative expression amount (fold) Control group (no additives) 1 ± 0.2 1 ± 0.02 Compound of Formula 1
(2 ㎍/mL) 1.61 ± 0.18 1.26 ± 0.12 Compound of Formula 1
(5 ㎍/mL) 2.42 ± 0.08 1.91 ± 0.3

As shown in Table 2, the compound of Formula 1 was found to increase the expression of antibacterial peptides LL37 and hBD2 in skin keratinocytes in a concentration-dependent manner compared to the untreated group.

<Example 3> Evaluation of skin moisture retention ability

In order to evaluate the moisturizing effect of the compound of Formula 1, a certain amount (2 mg/cm2) of the nutritional cream of Formulation Example 2 below was applied to the inside of the lower arm of 30 healthy adults in a constant temperature and humidity room at 22°C and 50% relative humidity. Before application, The moisture content of the skin was measured 1 hour after application and 2 hours after application. In the case of the control group, a nutritional cream containing the same amount of ethanol instead of the compound of Formula 1 was prepared and applied in the same way, and normal skin without any treatment was used as the untreated group. Water retention capacity was measured by measuring the electrical conductivity of the skin surface using a Corneometer (Courage + Khazaka, GmbH, Germany).

unit
(corneometer units)(n=30) Nutritional cream from Jeje Example 2 control group
Nutrition cream Untreated group Before application 50 51 56 1 hour after application 90 79 55 2 hours after application 85 62 54

As shown in Table 3, the nutritional cream containing the compound of Formula 1 increased the moisture content of human skin and showed excellent moisture retention ability even after 2 hours of application.

<Example 4> Evaluation of hyaluronic acid concentration

Human dermal fibroblasts were cultured at 1 × 10 ⁵ cells/mL in DMEM medium containing 10% fetal bovine serum, distributed at 500 ㎕ each in 24-well plates, and cultured for 18 hours. It was washed twice with DMEM medium containing no bovine serum, and the compound of Formula 1 was added at the following concentrations. As a negative control, the same amount of ethanol was treated, and as a positive control, 100 μg/mL of glucosamine hydrochloride (Sigma-aldrich), which is known to promote hyaluronic acid production, was treated. After culturing for 24 hours, the cell culture fluid was recovered and the hyaluronic acid concentration was measured using a Hyaluronan ELISA kit (R&D Systems).

In addition, the hyaluronic acid concentration of the experimental group treated with the compound of Formula 1 was quantified based on the hyaluronic acid concentration of the no-added control group (100%) and is shown in Table 4.

Sample (n=4) Hyaluronic acid relative concentration (%) Control group (no additives) 100 Glucosamine (100 μg/mL) 153 Compound of Formula 1 (5 μg/mL) 151 Compound of Formula 1 (10 μg/mL) 167

As shown in Table 4, the compound of Formula 1 increased hyaluronic acid production in human-derived fibroblasts.

<Example 5> Evaluation of expression level of hyaluronic acid synthase

Human dermal fibroblasts were cultured at 1 × 10 ⁶ cells/ml in DMEM medium containing 10% fetal bovine serum, distributed at 3000 ㎕ in 6-well plates, and cultured for 18 hours. Washed twice with DMEM medium without bovine serum. The compound of Formula 1 was treated at the following concentrations for 24 hours. Cells were recovered, total RNA was extracted using the RNeasy mini kit (Qiagen), quantified, and 1 μg of RNA was reverse transcribed using the GeneAmp ^® RNA PCR kit (Applied Biosystems). Reverse transcription reaction was performed using Mycycler ^® PCR instrument (Biorad).

Afterwards, quantitative real-time PCR was performed using 2 ㎕ of the reverse transcription reaction solution and Taqman ^® probes (Invitrogen, USA, HS02511055_S1, HS03929097_g1) of HAS-2, HAS-3, and GAPDH (Glyceraldehyde-3-phosphate dehydrogenase). Real-time PCR was performed using the CFX96 Touch ^TM Real-Time PCR Detection System device (Biorad). The experimental results obtained through real-time PCR were calculated and expressed by the method described in Livak KJ et al. ( Methods , 2001, 25, 402-408) based on the housekeeping gene GAPDH.

In addition, using the mRNA expression level of the negative control group as a standard (100%), the mRNA expression level of the experimental group treated with the compound of Formula 1 was quantified and shown in Table 5.

sample HAS-2 expression level (%) HAS-3 expression level (%) Control group (no additives) 100 100 Glucosamine
(100 ㎍/mL) 126 132 Compound of Formula 1
(5 ㎍/mL) 122 134 Compound of Formula 1
(10 ㎍/mL) 134 148

As shown in Table 5, it was found that the compound of Formula 1 increased the production of hyaluronic acid by increasing the mRNA expression level of hyaluronic acid synthase.

<Example 6> Evaluation of skin elasticity improvement

To evaluate the effect of the compound of Formula 1 on improving skin elasticity, the nutritional cream of Formulation Example 2 below was applied to the neck of 30 healthy adults in a constant temperature and humidity room at 22°C and 50% relative humidity for 4 weeks. After application, the elasticity before and after was measured to compare and evaluate the effect of improving skin elasticity.

The subject was to use a nutritional cream containing the compound of Chemical Formula 1 on the left side of the subject's neck, and a nutritional cream containing the same amount of ethanol instead of the compound of Chemical Formula 1 on the right side of the neck, twice a day in the morning and evening for 4 weeks after washing the face. Afterwards, the degree of improvement in elasticity in each area was measured using a skin elasticity meter (Cutometer, C+K, Germany). For the degree of improvement, Ur/Uf was used as a parameter.

Sample (n=30) Nutritional cream from Jeje Example 2 Nutritional cream in the control group Elasticity improvement rate (%) 25.13 5.6

As shown in Table 6, the compound of Formula 1 improved the elasticity of human skin and did not show any side effects.

Below, examples of various preparations containing the compound of Formula 1 as an active ingredient are described. However, the present invention is not limited to these formulation examples, and may be formulated with various other formulations well known to those skilled in the art in addition to the formulation examples described below.

<Formulation Example 1> Preparation of pharmaceutical formulation

1. Production of powder

Ingredients: 0.001 g of compound of formula 1, 1 g of lactose

A powder was prepared by mixing the above ingredients and filling them in an airtight bubble.

2. Preparation of tablets

Ingredients: 0.2 mg of compound of formula 1, 100 mg of corn starch, 100 mg of lactose, 2 mg of magnesium stearate

After mixing the above ingredients, tablets were manufactured by tableting according to a conventional tablet manufacturing method.

3. Manufacturing of capsules

Ingredients: 0.2 mg of compound of formula 1, 100 mg of corn starch, 100 mg of lactose, 2 mg of magnesium stearate

After mixing the above ingredients, a capsule was prepared by filling a gelatin capsule according to a typical capsule manufacturing method.

4. Manufacturing of pills

Ingredients: 0.003 g of compound of formula 1, 1.5 g of lactose, 1 g of glycerin, 0.5 g of xylitol

After mixing the above ingredients, pills were prepared to weigh 4 g per pill according to a conventional method.

5. Preparation of granules

Ingredients: Compound of Formula 1 2 mg, soybean extract 50 mg, glucose 200 mg, starch 600 mg

After mixing the above ingredients, 100 mg of 30% ethanol was added to prepare granules, which were dried at 60°C and the dried granules were filled into a granule packaging bag.

<Formulation Example 2> Manufacturing of cosmetics

1. Manufacturing of flexible lotion (skin lotion)

A softening lotion containing the compound of Formula 1 as an active ingredient was prepared according to a conventional method as shown in the following composition.

Composition of softening lotion:

0.1% by weight of compound of formula 1, 1.0% by weight of beta-1,3-glucan, 2.0% by weight of butylene glycol, 2.0% by weight of propylene glycol, 0.1% by weight of carboxyvinyl polymer, 0.2% by weight of PEG-12 nonylphenyl ether, Polysorbate 80 0.4% by weight, ethanol 10.0% by weight, triethanolamine 0.1% by weight, preservative 0.05% by weight, colorant 0.05% by weight, fragrance 0.05% by weight, purified water to 100% by weight.

2. Manufacturing of nutritional lotion (milk lotion)

A nutritional lotion containing the compound of Formula 1 as an active ingredient was prepared according to a conventional method as shown below.

Composition of nourishing lotion:

0.1% by weight of compound of formula 1, 1.0% by weight of beta-1,3-glucan, 4.0% by weight of beeswax, 1.5% by weight of polysorbate 60, 1.5% by weight of sorbitan sesquioleate, 0.5% by weight of liquid paraffin, caprylic /Capric triglyceride 5.0% by weight, glycerin 3.0% by weight, butylene glycol 3.0% by weight, propylene glycol 3.0% by weight, carboxyvinyl polymer 0.1% by weight, triethanolamine 0.2% by weight, preservative 0.05% by weight, colorant 0.05% by weight, Fragrance 0.05% by weight, purified water to 100% by weight

3. Manufacturing of nutritional cream

A nutritional cream containing the compound of Formula 1 as an active ingredient was prepared according to a conventional method as follows.

Composition of nourishing cream:

0.2% by weight of compound of formula 1, 5.0% by weight of beta-1,3-glucan, 10.0% by weight of beeswax, 1.5% by weight of polysorbate 60, 2.0% by weight of PEG 60 hydrogenated castor oil, 0.5% by weight of sorbitan sesquioleate , liquid paraffin 10.0% by weight, squalane 5.0% by weight, caprylic/capric triglyceride 5.0% by weight, glycerin 5.0% by weight, butylene glycol 3.0% by weight, propylene glycol 3.0% by weight, triethanolamine 0.2% by weight, preservative 0.05% Weight%, pigment 0.05% by weight, fragrance 0.05% by weight, purified water to 100% by weight

4. Manufacturing of massage cream

A massage cream containing the compound of Formula 1 as an active ingredient was prepared according to a conventional method with the following composition.

Composition of massage cream:

0.1% by weight of the compound of Formula 1, 3.0% by weight of beta-1,3-glucan, 10.0% by weight of beeswax, 1.5% by weight of polysorbate 60, 2.0% by weight of PEG 60 hydrogenated castor oil. Sorbitan sesquioleate 0.8% by weight, liquid paraffin 40.0% by weight, squalane 5.0% by weight, caprylic/capric triglyceride 4.0% by weight, glycerin 5.0% by weight, butylene glycol 3.0% by weight, propylene glycol 3.0% by weight, Triethanolamine 0.2% by weight, preservative 0.05% by weight, colorant 0.05% by weight, fragrance 0.05% by weight, purified water to 100% by weight

5. Preparation of packs

A pack containing the compound of Formula 1 as an active ingredient was prepared according to a conventional method with the following composition.

Composition of pack:

0.2% by weight of compound of formula 1, 1.0% by weight of beta-1,3-glucan, 13.0% by weight of polyvinyl alcohol, 0.2% by weight of sodium carboxymethyl cellulose, 5.0% by weight of glycerin, 0.1% by weight of allantoin, 6.0% by weight of ethanol, blood Easy-12 nonylphenyl ether 0.3% by weight, polysorbate 60 0.3% by weight, preservative 0.05% by weight, colorant 0.05% by weight, fragrance 0.05% by weight, purified water to 100% by weight

<Formulation Example 3> Preparation of external skin preparation

1. Preparation of gel

A gel containing the compound of Formula 1 as an active ingredient was prepared according to a conventional method with the following composition.

Composition of the gel:

0.1% by weight of compound of formula 1, 0.1% by weight of beta-1,3-glucan, 0.05% by weight of sodium ethylenediamine acetate, 5.0% by weight of glycerin, 0.3% by weight of carboxyvinyl polymer, 5.0% by weight of ethanol, PEG-60 cured caster Free 0.5% by weight, triethanolamine 0.3% by weight, preservative 0.05% by weight, colorant 0.05% by weight, fragrance 0.05% by weight, purified water to 100% by weight

2. Preparation of ointment

An ointment containing the compound of Formula 1 as an active ingredient was prepared according to a conventional method with the following composition.

Composition of ointment:

0.5% by weight of compound of formula 1, 10.0% by weight of beta-1,3-glucan, 10.0% by weight of beeswax, 5.0% by weight of polysorbate 60, 2.0% by weight of PEG 60 hydrogenated castor oil, 0.5% by weight of sorbitan sesquioleate , Vaseline 5.0% by weight, liquid paraffin 10.0% by weight, squalane 5.0% by weight, shea butter 3.0% by weight, caprylic/capric triglyceride 5.0% by weight, glycerin 10.0% by weight, propylene glycol 10.2% by weight, triethanolamine 0.2% by weight %, preservative 0.05% by weight, pigment 0.05% by weight, fragrance 0.05% by weight, purified water to 100% by weight

3. Manufacturing of drugs for topical administration (gel ointment)

A gel ointment containing the compound of Formula 1 as an active ingredient was prepared according to a conventional method with the following composition.

Composition of gel ointment:

0.5% by weight of compound of formula 1, 10.0% by weight of beta-1,3-glucan, 1.5% by weight of polyacrylic acid (Carbopol 940), 5.0% by weight of isopropanol, 25.0% by weight of hexylene glycol, 1.7% by weight of triethanolamine, deionized water to 100% by weight

4. Manufacturing of drugs for topical administration (patch)

A patch containing the compound of Formula 1 as an active ingredient was prepared according to a conventional method as shown in the following composition.

Composition of patch:

0.5% by weight of compound of formula 1, 3.0% by weight of beta-1,3-glucan, 20.0% by weight of hexylene glycol, 0.7% by weight of diethylamine, 1.0% by weight of polyacrylic acid (Carbopol 934P), 0.1% by weight of sodium sulfite, poly 1.0% by weight of oxyethylene lauryl ether (E.O=9), 1.0% by weight of polyhydroxyethylene cetylstearyl ether (Cetomacrogol 1000), 2.5% by weight of viscous paraffin oil, caprylic acid ester/capric acid ester (Cetiol LC) ) 2.5% by weight, polyethylene glycol 400 3.0% by weight, deionized water to 100% by weight

Claims (6)

delete delete A cosmetic composition for moisturizing skin containing a compound represented by the following formula (1) as an active ingredient:
[Formula 1]

A food composition for improving skin moisturization comprising a compound represented by the following formula (1) as an active ingredient:
[Formula 1]

A skin external composition for moisturizing skin containing a compound represented by the following formula (1) as an active ingredient:
[Formula 1]
delete