Rhamnose derivative

Abstract

The present invention relates to a rhamnose derivative or a pharmaceutically or cosmetically acceptable salt thereof, and a pharmaceutical or cosmetic composition comprising same. The rhamnose derivative exhibits wound healing effects, as well as skin elasticity enhancement, wrinkle alleviation, skin barrier strengthening, and skin moisturizing effects, and thus can be effectively used in the pharmaceutical and cosmetics fields.

Description

Rhamnose derivatives

[0001] The present invention relates to novel rhamnose derivatives or acceptable salts thereof, and novel pharmaceutical or cosmetic uses thereof.

[0002]

[0003] Rhamnose is a naturally occurring monosaccharide, an aldose with six carbon atoms, and a deoxy sugar in which one hydroxyl group is replaced by a hydrogen. Its chemical formula is C6H 12 It is O5 and can be classified as methylpentose or 6-deoxyhexose. L-rhamnose is moderately soluble in water, with a solubility of 0.1 g / mL (Sigma-Aldrich).

[0004] The skin is the organ with the largest surface area in our body. Directly exposed to the external environment, the skin serves the function of protecting the body from various harmful factors. However, as the skin becomes increasingly damaged by increasing age, ultraviolet radiation, external pollutants, and stress, and as its protective function against these factors weakens, its ability to protect and proliferate cells declines. In this case, the body initiates a skin regeneration process for the damaged skin. Cell division and migration occur, and through these actions, the wound heals. Therefore, wound healing measurements that analyze cell division and migration are associated with skin regeneration (Gerontology (2013) 59 (2): 159-164. Age-Related Aspects of Cutaneous Wound Healing: A Mini-Review) (Pharmaceuticals 2023, 16(2), 235, Skin Wound Healing and Anti-Wrinkle-Promoting In Vitro Biological Activities of Caragana sinica Flower Absolute and Its Chemical Composition). Furthermore, for cells to divide, their functions and activities must operate normally. Therefore, analyzing the relative number of cells is associated with cell activity (Aging Clin Exp Res1, 3-15 (1989). Cell proliferation, cell death and aging).

[0005] Human skin consists of the epidermis and the dermis. Fibroblasts exist in the dermis and produce collagen, a major component of the extracellular matrix. Collagen is an important protein that accounts for approximately 30% of the total body protein weight; it has a rigid triple helix structure and supports the mechanical rigidity and intercellular adhesion of the skin. Collagen decreases due to natural aging or photoaging caused by ultraviolet (UV) irradiation, and this is known to be directly related to the formation of wrinkles on the skin.

[0006] UV promotes collagen degradation by stimulating the expression of MMP-1, one of the collagen-degrading enzymes. The reduction of collagen caused by MMP-1 decreases skin elasticity and promotes wrinkle formation. Therefore, demonstrating efficacy in reducing MMP-1 expression is associated with wrinkle improvement (Photochemistry and Photobiology, 2003, 78(1): 43-48, Matrix Metalloproteinase-1 is the Major Collagenolytic Enzyme Responsible for Collagen Damage in UV-irradiated Human Skin).

[0007] The epidermis is located at the outermost layer and performs a defense against various external physical and chemical stimuli, as well as a protective function that prevents the excessive loss of moisture from the body. These defense and protective functions are made possible by the normal formation and maintenance of the stratum corneum, which is composed of keratinocytes. Keratinocytes are cells formed through a gradual process in which cells that have continuously proliferated in the lowest layer of the epidermis migrate to the stratum corneum.

[0008] These keratinocytes undergo repeated processes of differentiation or keratinization. Through this process, old keratinocytes are shed from the skin, and new keratinocytes rising from the lowest layer take over their function. During this process, keratinocytes form moisturizing factors and intercellular lipids, enabling the stratum corneum to function as a protective barrier against external factors.

[0009] Tight junctions (TJs) play a crucial role in this function as a barrier. Tight junctions are intercellular contact structures that seal the spaces between individual cells of epithelial sheets or interstratifying epithelia, such as the epidermis, enabling the collective separation of tissue compartments. Representative molecules constituting tight junctions, such as Claudin and Occludin, bind to actin filaments on the outer walls of skin cells, serving as connection points between cells to strengthen the binding capacity of the skin barrier. This enhanced binding force prevents the invasion of pathogens from the outside of the skin and reduces moisture loss from within (Ther Deliv. 2012 Nov;3(11):1297-327. Tight junctions in skin: new perspectives) (Journal of Investigative Dermatology (2014) 135, e29. Assessing the In Vivo Epidermal Barrier in Mice: Dye Penetration Assays).

[0010] Hyaluronic acid, produced by keratinocytes, is a moisturizing factor classified as a type of glycosaminoglycan. It is a high-molecular-weight polysaccharide consisting of repeating glucuronic acid and N-acetylglucosamine residues. As a major component of the extracellular matrix, hyaluronic acid is closely related to skin moisture content. Hyaluronic acid has the property of gelling by binding with large amounts of water, and functionally, it is associated with moisture retention and the maintenance of skin elasticity. Hyaluronic acid synthase is a hyaluronic acid synthesis enzyme present in cells, such as HAS1, HAS2, and HAS3, and hyaluronic acid of different lengths is formed depending on the type. Hyaluronic acid produced by cells acts as a humectant, attracting moisture and increasing the moisture content in the skin (Curr Med Chem. 2009;16(14):1718-45. Role, metabolism, chemical modifications and applications of hyaluronan).

[0011] In addition, aquaporins are pumps for the transport of water and glycerol expressed in skin cells, and they play a role in increasing moisture levels through even delivery to the skin layers (Journal of Investigative Dermatology (2008) 128, 2145-2151; Roles of Aquaporin-3 in the Epidermis).

[0012]

[0013] In the present invention, a novel rhamnose derivative that did not previously exist was synthesized. Then, the present invention was completed by preparing a pharmaceutical composition and a cosmetic composition containing the rhamnose derivative and confirming the skin-improving effect thereof.

[0014]

[0015] The present invention aims to provide a novel rhamnose derivative or a pharmaceutically or cosmetically acceptable salt thereof.

[0016] In addition, the present invention aims to provide a pharmaceutical composition for wound treatment comprising a rhamnose derivative or a pharmaceutically acceptable salt thereof.

[0017] In addition, the present invention aims to provide a cosmetic composition for improving skin elasticity or wrinkles, strengthening the skin barrier, or moisturizing the skin, comprising a derivative or a cosmetically acceptable salt thereof.

[0018]

[0019] The present invention provides a rhamnose derivative represented by Formula 1, or a salt thereof acceptable for pharmaceutical or cosmetic use.

[0020]

[0021] [Chemical Formula 1]

[0022]

[0023] In the above chemical formula 1,

[0024] R1 is -(CH2)n-R2 or -(CH) m -(R3)2 and,

[0025] R2 is a hydroxyl group, carboxyl group, amide group, amine group, ester group, nitrile group, or acetylmorpholine, and

[0026] R3 is -COOH or -COO - M + And,

[0027] M is Na, K, or Li, and

[0028] n and m are each independently integers from 1 to 8.

[0029]

[0030] In addition, the present invention provides a pharmaceutical composition for wound treatment comprising the aforementioned rhamnose derivative or a pharmaceutically acceptable salt thereof.

[0031]

[0032] In addition, the present invention provides a cosmetic composition for improving skin elasticity or wrinkles, strengthening the skin barrier, or moisturizing the skin, comprising the aforementioned rhamnose derivative or a cosmetically acceptable salt thereof.

[0033]

[0034] The novel rhamnose derivative according to the present invention can provide wound healing effects, improve skin elasticity, improve skin wrinkles, strengthen the skin barrier, or provide skin moisturizing effects, so it can be usefully used in fields such as pharmaceuticals and cosmetics.

[0035]

[0036] The present invention will be described in more detail below.

[0037] Each description and embodiment disclosed herein may be applied to each other description and embodiment. That is, all combinations of the various elements disclosed herein fall within the scope of the invention. Furthermore, the scope of the invention is not to be limited by the specific descriptions provided below.

[0038] When a part is said to "include" a certain component, this means that, unless specifically stated otherwise, it does not exclude other components but rather allows for the inclusion of additional components.

[0039] The present invention provides a rhamnose derivative represented by the following chemical formula 1, or a pharmaceutically or cosmetically acceptable salt thereof.

[0040]

[0041] [Chemical Formula 1]

[0042]

[0043] In the above chemical formula 1,

[0044] R1 is -(CH2)n-R2 or -(CH) m -(R3)2 and,

[0045] R2 is a hydroxyl group, carboxyl group, amide group, amine group, ester group, nitrile group, or acetylmorpholine, and

[0046] R3 is -COOH or -COO - M + And,

[0047] M is Na, K, or Li, and

[0048] n and m are each independently integers from 1 to 8.

[0049] In the present invention, “the pharmaceutically or cosmetically acceptable salts thereof” may include salts prepared by adding a base to a rhamnose derivative. The addition salts available for preparing the pharmaceutically or cosmetically acceptable salts include, but are not limited to, one or more salts selected from the group consisting of sodium, potassium, calcium, ammonium, magnesium, and organic aminos.

[0050] In one embodiment, R2 may be a hydroxyl group, a carboxyl group, an amide group, a nitrile group, or acetylmorpholine.

[0051] In one specific example, R3 is -COO - M + It can be, specifically, it can be -COONa.

[0052] In addition, in one embodiment, n may be an integer from 1 to 4, specifically 1, 2, or 3.

[0053] In the present invention, the rhamnose derivative represented by Chemical Formula 1 may be one of the compounds represented by Chemical Formulas 2 to 8 below.

[0054] [Chemical Formula 2]

[0055]

[0056] [Chemical Formula 3]

[0057]

[0058] [Chemical Formula 4]

[0059]

[0060] [Chemical Formula 5]

[0061]

[0062] [Chemical Formula 6]

[0063]

[0064] [Chemical Formula 7]

[0065]

[0066] [Chemical Formula 8]

[0067]

[0068]

[0069] In addition, the present invention provides a pharmaceutical composition for wound treatment comprising the aforementioned rhamnose derivative or a pharmaceutically acceptable salt thereof.

[0070] In this invention, "wound" encompasses all damage to a living organism and refers to a state in which a living organism is damaged; it is also referred to as a wound.

[0071] The "wound healing" process is divided into the acute, repair, and scarring phases. The acute phase, also known as the exudative phase, is a stage where a series of reactions occur to remove tissue destruction or foreign substances from the damaged area; this is accompanied by inflammatory and blood coagulation responses. The repair phase, also known as the proliferative phase, is a stage where new blood vessels are formed and the damaged area expands to facilitate recovery. During this phase, active cell proliferation or the active synthesis of collagen and proteoglycans—intercellular substances found in granulation tissue, a type of connective tissue—occurs, allowing epidermal cells to gain mobility and proliferate through cell division to regenerate epidermal tissue. The scarring phase is a stage where active cell proliferation slows down, and the physical strength of the damaged area increases as collagen fibers cross-link. Ultimately, the vascular system atrophies, and tissue distinct from the surrounding normal tissue takes its place at the damaged site. Wound healing occurs through the repetition of these stages.

[0072] In one embodiment, the wound may be a wound in which the epidermis; dermis; epidermis and dermis; or the epidermis, dermis and subcutaneous fat layer of the skin is damaged. Additionally, the wound may include cuts, incisions (e.g., surgical incisions), abrasions, lacerations or lacerations, fractures, contusions, burns, or amputations.

[0073] The above wound may be selected from a group consisting of chronic wounds, acute wounds, surgical wounds, orthopedic wounds, traumatic wounds, combat wounds, and combinations thereof, or may be a wound caused by other diseases. In this case, the disease may be selected from fibrosis, diabetes mellitus, diabetic ulcers, autoimmune skin diseases, abrasions, lacerations, incisions, contusions, bruises, punctures, excisions, burns, ulcers, pressure sores, or combinations thereof.

[0074] In this invention, “treatment” refers to any act of alleviating, reducing, stopping, or reversing desired symptoms by administering the composition of this invention.

[0075] The above “pharmaceutical composition” refers to a composition used for the purpose of diagnosing, treating, alleviating, managing, or preventing diseases in animals, including humans.

[0076] In one embodiment, in a wound healing assay analysis in which a rhamnose derivative was administered in the above content range, it was confirmed that the rhamnose derivative exhibited an enhanced wound healing effect through significantly enhanced growth activity of skin fibroblasts.

[0077] In one embodiment, a rhamnose derivative, or a pharmaceutically acceptable salt thereof, may be included in an amount of 0.00001 wt% to 20 wt%, 0.0001 wt% to 20 wt%, 0.001 wt% to 20 wt%, 0.005 wt% to 20 wt%, 0.01 wt% to 20 wt%, 0.05 wt% to 20 wt%, 0.1 wt% to 20 wt%, 0.00001 wt% to 10 wt%, 0.0001 wt% to 10 wt%, 0.001 wt% to 10 wt%, 0.005 wt% to 10 wt%, 0.01 wt% to 10 wt%, 0.05 wt% to 10 wt%, or 0.1 wt% to 10 wt% based on the total weight of the pharmaceutical composition. Specifically, it may be included in an amount of 0.001 to 5 weight%, 0.01 to 5 weight%, 0.1 to 5 weight%, 0.1 to 4 weight%, 0.1 to 3 weight%, 0.1 to 2 weight%, or 0.1 to 1 weight%. Excellent wound healing effects may be achieved within the above content range. If included below the content range, cell activity induction at the wound site may be low, resulting in no wound healing effect or a slow healing speed; if included above the above content range, problems such as skin irritation may occur.

[0078] The composition of the present invention may be administered in a pharmaceutically effective amount. The pharmaceutically effective amount refers to an amount sufficient to treat a disease with a reasonable benefit / risk ratio applicable to medical treatment and that does not cause side effects. The effective dose level may be determined based on factors including the patient's health status, type and severity of the disease, drug activity, sensitivity to the drug, method of administration, time of administration, route of administration and elimination rate, duration of treatment, drugs used in combination or concurrently, and other factors well known in the medical field. Specifically, it may be 0.1 mg / kg to 100,000 mg / kg per day, more specifically 1 mg / kg to 10,000 mg / kg. The pharmaceutical composition of the present invention may be administered once a day or divided into several doses. Therefore, the above dosage does not limit the scope of the present invention in any way.

[0079] The above composition may be administered to mammals such as rats, mice, livestock, and humans via various routes, such as parenteral or oral, and all modes of administration are expected, for example, by oral, rectal or intravenous, intramuscular, subcutaneous, intrauterine dura mater, or intracerebroventricular injection.

[0080] The pharmaceutical composition of the present invention may further include pharmaceutically acceptable carriers, excipients, diluents, and / or auxiliary components depending on its formulation, method of use, and purpose of use.

[0081] In addition, the present invention provides a quasi-drug composition for wound treatment comprising the aforementioned rhamnose derivative or a pharmaceutically acceptable salt thereof.

[0082] In the present invention, "quasi-drug" refers to articles used for the purpose of diagnosing, treating, improving, alleviating, managing, or preventing diseases in humans or animals, and which have a milder effect than pharmaceuticals. For example, according to the Pharmaceutical Affairs Act, quasi-drugs are defined as products excluding those used for pharmaceutical purposes, and may be products used for the treatment or prevention of diseases in humans or animals, or products that have a mild effect on the human body or do not act directly on it. For example, they may be selected from the group consisting of disinfectants, detergents, kitchen detergents, cleaning detergents, wet wipes, detergents, soaps, hand washes, hair detergents, hair softeners, and ointments, but are not limited thereto.

[0083]

[0084] In addition, the present invention provides a cosmetic composition for improving skin elasticity or wrinkles comprising the aforementioned rhamnose derivative or a cosmetically acceptable salt thereof.

[0085] In addition, the present invention provides a cosmetic composition for skin barrier strengthening or skin moisturizing comprising a rhamnose derivative or a cosmetically acceptable salt thereof.

[0086] In the present invention, “skin elasticity enhancement” means alleviating the degree of sagging or stretching of the skin, and furthermore, said elasticity may mean maintaining the elasticity of the skin in a state where collagen is sufficiently present.

[0087] In the present invention, “wrinkle improvement” refers to inhibiting or hindering the formation of wrinkles on the skin, or alleviating wrinkles that have already formed.

[0088] In the present invention, “skin barrier strengthening” means restoring the barrier function of the stratum corneum located on the outermost part of the skin.

[0089] In the present invention, “skin moisturization” refers to increasing moisture in the skin and maintaining a moist state. The skin moisturizing effect can also help improve wrinkles and increase skin elasticity.

[0090] The rhamnose derivative of the present invention inhibits the expression of the MMP-1 gene, which is involved in the collagen degradation process in the dermis of the skin, thereby enhancing skin elasticity and inhibiting wrinkle formation. In addition, it can exhibit skin barrier strengthening and moisturizing effects by enhancing the expression of tight junction genes responsible for the binding between epidermal cells and enhancing the expression of HAS3 (Hyaluronic acid Synthase 3) genes that induce hyaluronic acid synthesis.

[0091] In one embodiment, a rhamnose derivative, or a cosmetically acceptable salt thereof, may be included in an amount of 0.00001 wt% to 20 wt%, 0.0001 wt% to 20 wt%, 0.001 wt% to 20 wt%, 0.005 wt% to 20 wt%, 0.01 wt% to 20 wt%, 0.05 wt% to 20 wt%, 0.1 wt% to 20 wt%, 0.00001 wt% to 10 wt%, 0.0001 wt% to 10 wt%, 0.001 wt% to 10 wt%, 0.005 wt% to 10 wt%, 0.01 wt% to 10 wt%, 0.05 wt% to 10 wt%, or 0.1 wt% to 10 wt% based on the total weight of the cosmetic composition. Specifically, it may be included in an amount of 0.001 to 5 weight%, 0.01 to 5 weight%, 0.1 to 5 weight%, 0.1 to 4 weight%, 0.1 to 3 weight%, 0.1 to 2 weight%, or 0.1 to 1 weight%. If a rhamnose derivative, or a cosmetically acceptable salt thereof, is included in an amount lower than the above range, the desired effect cannot be obtained, and if it is included in an amount higher than the above range, a problem may arise that it may cause skin irritation.

[0092] The cosmetic composition according to the present invention may be provided in any formulation suitable for topical application and may be prepared in a formulation selected from the group consisting of solutions, topical ointments, creams, foams, nourishing lotions, softening lotions, packs, softening waters, emulsions, makeup bases, essences, soaps, liquid cleansers, bath additives, sunscreen creams, sun oils, suspensions, emulsions, pastes, gels, lotions, powders, soaps, surfactant-containing cleansing oils, powder foundations, emulsion foundations, wax foundations, patches, and sprays, but is not limited thereto.

[0093] In addition, the cosmetic composition of the present invention may further include one or more cosmetically acceptable carriers that are incorporated into general skin cosmetics, and may appropriately incorporate, for example, oils, water, surfactants, moisturizers, lower alcohols, thickeners, chelating agents, pigments, preservatives, fragrances, etc., as conventional ingredients, but is not limited thereto.

[0094] The above carrier can be introduced in an amount commonly used in the fields of cosmetics or dermatology.

[0095]

[0096] In addition, the present invention relates to the wound healing use of the aforementioned rhamnose derivative or a pharmaceutically acceptable salt thereof.

[0097] In addition, the present invention relates to the use of the aforementioned rhamnose derivative, or a cosmetically acceptable salt thereof, for improving skin elasticity or wrinkles.

[0098] In addition, the present invention relates to the use of the aforementioned rhamnose derivative, or a cosmetically acceptable salt thereof, for strengthening the skin barrier or moisturizing the skin.

[0099]

[0100] In addition, the present invention relates to a wound treatment method comprising the step of applying a pharmaceutical composition containing an effective amount of the aforementioned rhamnose derivative or a pharmaceutically acceptable salt thereof to the skin of a subject in need thereof.

[0101] In addition, the present invention relates to a method for improving skin elasticity or wrinkles, comprising the step of applying a cosmetic composition containing an effective amount of the aforementioned rhamnose derivative or a cosmetically acceptable salt thereof to the skin of a subject who requires it.

[0102] In addition, the present invention relates to a method for strengthening the skin barrier or moisturizing the skin, comprising the step of applying a cosmetic composition containing an effective amount of the aforementioned rhamnose derivative or a cosmetically acceptable salt thereof to the skin of a subject who requires it.

[0103]

[0104] The present invention will be described in detail below through examples. The following examples are merely illustrative of the present invention and do not limit the scope of the present invention to the following examples. These examples are provided to ensure that the disclosure of the present invention is complete and to fully inform those skilled in the art of the scope of the invention, and the present invention is defined only by the scope of the claims.

[0105]

[0106] Preparation Example 1. Synthesis of 2-((3,4,5-trihydroxy-6-methyltetrahydro-2H-pyran-2-yl)oxy)acetic acid

[0107] 1 g of pentaacetylamnose was dispersed in 6 mL of dichloromethane (DCM), and 0.5 g of methyl glycolate was dissolved in 6 mL of DCM and added. After cooling the mixture to 0°C, 0.6 mL of boron trifluoride etherate (BF3-Et2O) solution was slowly added. The reaction mixture was heated to 30°C and stirred at this temperature for 6 hours. After the reaction was complete, 5 mL of distilled water and 5 mL of DCM were added to the reaction solution to stop the reaction, and the organic phase was separated. The organic phases were combined and sequentially washed with saturated sodium bicarbonate (NaHCO3). After drying with anhydrous sodium sulfate (Na2SO4), the solvent was removed by vacuum concentration. The crude product was purified by silica gel column chromatography (PE / EA=3:1, v / v) to obtain a clean intermediate.

[0108] 10 ml of sodium methoxide (NaOMe) solution (5 mol / L in methanol (MeOH)) was added, the intermediate was dissolved, and the mixture was stirred at room temperature for 30 minutes. Then, the solution was neutralized with Amberlite IRA-120 resin and filtered to remove the solvent.

[0109] 1 H NMR (DO, 400 MHz): 1.07(3H), 3.61(1H), 3.67(1H), 3.81(2H), 3.97(2H), 4.77(1H)

[0110]

[0111] Preparation Example 2. Synthesis of sodium 2-((-3,4,5-trihydroxy-6-methyltetrahydro-2H-pyran-2-yl)oxy)malonate

[0112] 1 g of pentaacetylamnose was dispersed in 6 mL of DCM, and 1.2 g of 2-hydroxymalonic acid diethyl ester was dissolved in 6 mL of DCM and added. After cooling the mixture to 0 °C, 0.6 mL of BF3-Et2O solution was slowly added. The reaction mixture was heated to 30 °C and stirred at this temperature for 6 hours. After the reaction was complete, 5 mL of distilled water and 5 mL of DCM were added to the reaction solution to stop the reaction, and the organic phase was separated. The organic phases were combined and washed sequentially with saturated NaHCO3. After drying with anhydrous Na2SO4, the solvent was removed by vacuum concentration. The crude product was purified by silica gel column chromatography (PE / EA=3:1, v / v) to obtain a clean intermediate.

[0113] 10 ml of NaOMe solution (5 mol / L in MeOH) was added, the intermediate was dissolved, and the mixture was stirred at room temperature for 30 minutes. Then, the solution was neutralized with Amberlite IRA-120 resin and filtered to remove the solvent.

[0114] 1H NMR (DO, 600 MHz): 1.27(3H), 3.44(1H), 3.74(1H), 3.95 (1H), 4.10(1H), 4.45(1H), 4.78(H)

[0115] 13 C NMR (DO, 600 MHz): 16.6, 69.2, 70.1, 72.1, 79.7, 99.6, 174.3

[0116]

[0117] Preparation Example 3. 2-((3,4,5-trihydroxy-6-methyltetrahydro-2H-pyran-2-yl)oxy)acetamide

[0118] 1 g of pentaacetylamnose was dispersed in 6 mL of DCM, and 0.5 g of methyl glycolate was dissolved in 6 mL of DCM and added. After cooling the mixture to 0°C, 0.6 mL of BF3-Et2O solution was slowly added. The reaction mixture was heated to 30°C and stirred at this temperature for 6 hours. After the reaction was complete, 5 mL of distilled water and 5 mL of DCM were added to the reaction solution to stop the reaction, and the organic phase was separated. The organic phases were combined and washed sequentially with saturated NaHCO3. After drying with anhydrous Na2SO4, the solvent was removed by vacuum concentration. The crude product was purified by silica gel column chromatography (PE / EA=3:1, v / v) to obtain a clean intermediate.

[0119] Treatment was carried out at 25°C with 10 mL of ammonia solution in MeOH until conversion was complete. Then, the solvent was evaporated under reduced pressure, and the crude residue was washed three times with 10 mL of ethyl acetate (AcOEt) to obtain a pure product.

[0120] 1 H NMR (DO, 600 MHz): 1.30(3H), 3.46(1H), 3.73(1H), 3.85(1H), 4.10(1H), 4.11(1H), 4.22(1H), 4.85(H)

[0121]

[0122] Experimental Example 1: Skin regeneration efficacy by rhamnose derivative

[0123] To confirm the skin regeneration effect following treatment with a rhamnose derivative, a wound healing assay was performed.

[0124] First, human dermal fibroblasts (CRL-1635) were purchased from ATCC and used. The human dermal fibroblasts were cultured in DMEM (Dulbecco Modified Eagle Medium-GIBCO) medium containing 10% FBS (fetal bovine serum) and 1% Penicillin / Streptomycin at 37°C in a 5% CO2 incubator.

[0125] Next, transfer the cells to a 24-well plate at a rate of 2 x 10 4 Cells were dispensed per well and cultured for one day in a 37°C, 5% CO2 incubator. Subsequently, small wounds 1 mm wide were created using an SPLScar™ scratcher (SPL, 201925), and then 1,000 ppm LGSL1, 250 ppm LGSL2, and 250 ppm LGSL3 were added to the medium. In this case, LGSL1, LGSL2, and LGSL3 were rhamnose derivatives prepared in Preparation Examples 1, 2, and 3, respectively. As a control, the same concentration of rhamnose was added to the medium and cultured. After 24 hours of culture, the wound area was measured to confirm the wound healing effect.

[0126] The analysis results are shown in Tables 1 and 2.

[0127]

[0128] Group Wound Recovery Rate (%) Increase Compared to Rhamnose (%) Untreated 100.0 - Rhamnose 138.164 - LGSL 1161.550 16.93

[0129]

[0130] Group Wound Recovery Rate (%) Increase Compared to Rhamnose (%) Untreated 100.0 - Rhamnose 104.403 - LGSL 2115.896 11.01 LGSL 3110.467 5.81

[0131]

[0132] As shown in the table above, it can be confirmed that the rhamnose derivative according to the present invention exhibits a higher wound healing rate compared to rhamnose.

[0133]

[0134] Experimental Example 2: Efficacy of reducing wrinkle-associated gene expression by rhamnose derivative

[0135] We confirmed whether rhamnose derivatives affect the expression of MMP1, a gene involved in the collagen degradation process in the dermis of the skin.

[0136] Human skin fibroblasts CRL-1635 were cultured in DMEM medium containing 10% FBS and 1% Penicillin / Streptomycin at 37°C in a 5% CO2 incubator.

[0137] Next, transfer the cells to a 6-well plate at a rate of 3 x 10 5 Cells were seeded per well and cultured for one day at 37°C in a 5% CO2 incubator. After replacing the medium with phosphate-buffered bacon (PBS), the pressure was 20 mJ / cm². 2 After treating with UVB at a certain intensity, the rhamnose derivative according to the present invention and the control rhamnose were added to the medium at the concentrations specified in Table 3. Subsequently, the culture was carried out for one day under the same conditions as before. Then, total RNA was obtained using the AccuPrep Universal RNA Extraction Kit (Bionia, K-3140), and cDNA was synthesized using a cDNA synthesis kit (Philekorea). The gene expression level was analyzed by performing qPCR using the synthesized cDNA.

[0138] The results are shown in Table 3.

[0139]

[0140] Group Gene Expression (%) Untreated 100 Rhamnose 500 ppm 55 Rhamnose 1000 ppm 62 LGSL2 500 ppm 52 LGSL2 1000 ppm 68 LGSL3 500 ppm 40 LGSL3 1000 ppm 45

[0141]

[0142] As shown in the table above, it can be confirmed that the rhamnose derivatives according to the present invention have an efficacy of reducing wrinkle-associated gene expression equivalent to or greater than that of rhamnose. In particular, LGLS3 showed the most superior efficacy in reducing expression.

[0143]

[0144] Experimental Example 3: Cell activating efficacy by rhamnose derivative

[0145] To determine whether rhamnose derivatives affect cell activity, cell activity was confirmed by measuring the relative number of cells using a CCK-8 assay.

[0146] Human skin fibroblasts CRL-1635 were cultured in DMEM medium containing 10% FBS and 1% Penicillin / Streptomycin at 37°C in a 5% CO2 incubator.

[0147] Next, transfer the cells to a 96-well plate at a ratio of 1 x 10⁻⁶ 4 Cells were seeded per well and incubated for one day at 37°C in a 5% CO2 incubator. After replacing the medium with PBS, the setting was 20 mJ / cm². 2 After treating with UVB at a certain intensity, the rhamnose derivative according to the present invention and the control rhamnose were added to the medium at the concentrations according to Table 4. Then, the culture was carried out for one day under the same conditions as before. Subsequently, the relative number of cells was measured using the Cell Counting Kit-8 cell proliferation assay kit (Dojindo Molecular Technologies Inc.).

[0148] The results are shown in Table 4.

[0149]

[0150] Group Relative Cell Count (%) Untreated 100 Rhamnose 500 ppm 110 Rhamnose 1000 ppm 99 LGSL1 500 ppm 106 LGSL1 1000 ppm 106 LGSL2 500 ppm 133 LGSL2 1000 ppm 129 LGSL3 500 ppm 128 LGSL3 1000 ppm 127

[0151]

[0152] As shown in the table above, it can be confirmed that the rhamnose derivative according to the present invention has superior cell activity efficacy compared to rhamnose.

[0153]

[0154] Experimental Example 4: Skin barrier strengthening efficacy by rhamnose derivative

[0155] To determine whether rhamnose derivatives affect cell activity, skin barrier strengthening efficacy was confirmed by measuring tight junction genes (Occludin, Claudin4, TJP1), which are markers responsible for the binding between epidermal cells.

[0156] Human keratinocytes (HaCaT, Cytion 300493) were purchased from Cytion and used. The human keratinocytes were cultured in DMEM medium containing 10% FBS and 1% Penicillin / Streptomycin at 37°C in a 5% CO2 incubator.

[0157] Next, transfer the cells to a 6-well plate at a rate of 2 x 10 5Cells were dispensed into wells and cultured for one day in a 37°C, 5% CO2 incubator. The rhamnose derivative according to the present invention and the control rhamnose were added to the medium at the concentrations specified in Tables 5, 6, and 7. Subsequently, the cultures were incubated for one day under the same conditions as before. Then, total RNA was obtained using the AccuPrep Universal RNA Extraction Kit, and cDNA was synthesized using a cDNA synthesis kit. The gene expression levels were analyzed by performing qPCR using the synthesized cDNA.

[0158] The analysis results are shown in Tables 5, 6, and 7, respectively.

[0159]

[0160] Gene Group Relative Expression Level (%) Claudin 4 Untreated 100 Rhamnose 2000 ppm 169 LGSL1 2000 ppm 679

[0161]

[0162] Gene Group Relative Expression Level (%) Occludin Untreated 100 Rhamnose 200 ppm 86 LGSL2 200 ppm 174

[0163]

[0164] Gene Group Relative Expression Level (%) TJP1 Untreated 100 Rhamnose 200 ppm 95 LGSL3 200 ppm 131

[0165]

[0166] As shown in Tables 5, 6, and 7, it can be confirmed that the expression levels of Occludin, Claudin4, and TJP1 increase when the rhamnose derivative according to the present invention is used.

[0167] Through this, it can be confirmed that rhamnose derivatives have superior skin barrier strengthening efficacy compared to rhamnose.

[0168]

[0169] Experimental Example 5: Moisturizing Enhancement Efficacy by Rhamnose Derivative

[0170] To determine whether rhamnose derivatives affect cell activity, hyaluronic acid synthesis and intercellular water transport genes (HAS2 / 3, Aquaporin3) were measured to confirm moisturizing-enhancing efficacy.

[0171] Human keratinocyte Cytion 300493 was cultured in DMEM medium containing 10% FBS and 1% Penicillin / Streptomycin at 37°C in a 5% CO2 incubator.

[0172] Next, transfer the cells to a 6-well plate at a rate of 2 x 10 5 Cells were dispensed into wells and cultured for one day in a 37°C, 5% CO2 incubator. The rhamnose derivative according to the present invention and the control rhamnose were added to the medium at the concentrations specified in Tables 8, 9, and 10. Subsequently, the cultures were incubated for one day under the same conditions. Then, total RNA was obtained using the AccuPrep Universal RNA Extraction Kit, and cDNA was synthesized using a cDNA synthesis kit. The gene expression levels were analyzed by performing qPCR using the synthesized cDNA.

[0173] The analysis results are shown in Tables 8, 9, and 10.

[0174]

[0175] Gene Group Relative Expression Level (%) Hyaluronic acid Synthase 3 Untreated 100 Rhamnose 2000 ppm 130 LGSL1 2000 ppm 440

[0176]

[0177] Gene Group Relative Expression Level (%) Hyaluronic acid Synthase 2 Untreated 100 Rhamnose 2000 ppm 132 LGSL2 2000 ppm 169

[0178]

[0179] Gene Group Relative Expression Level (%) Aquaporin 3 Untreated 100 Rhamnose 200 ppm 113 LGSL3 200 ppm 145

[0180]

[0181] As shown in Tables 8, 9, and 10, it can be confirmed that the expression levels of Hyaluronic acid 2, Hyaluronic acid 3, and Aquaporin 3 increase when the rhamnose derivative according to the present invention is used.

[0182] Through this, it can be confirmed that rhamnose derivatives have superior moisturizing enhancement efficacy compared to rhamnose.

Claims

1. A rhamnose derivative represented by the following chemical formula 1, or a pharmaceutically or cosmetically acceptable salt thereof: [Chemical Formula 1] In the above chemical formula 1, R1 is -(CH2)n-R2 or -(CH) m -(R3)2 and, R2 is a hydroxyl group, carboxyl group, amide group, amine group, ester group, nitrile group, or acetylmorpholine, and R3 is -COOH or -COO - M + And, M is Na, K, or Li, and n and m are each independently integers from 1 to 8.

2. In Paragraph 1, A rhamnose derivative represented by Chemical Formula 1 is a rhamnose derivative that is a compound represented by Chemical Formulas 2 to 8 below, or a pharmaceutically or cosmetically acceptable salt thereof: [Chemical Formula 2] [Chemical Formula 3] [Chemical Formula 4] [Chemical Formula 5] [Chemical Formula 6] [Chemical Formula 7] [Chemical Formula 8] 3. A pharmaceutical composition for wound treatment comprising a rhamnose derivative according to claim 1, or a pharmaceutically acceptable salt thereof.

4. A cosmetic composition for improving skin elasticity or wrinkles comprising a rhamnose derivative according to claim 1, or a cosmetically acceptable salt thereof.

5. A cosmetic composition for skin barrier strengthening or skin moisturizing comprising a rhamnose derivative according to claim 1, or a cosmetically acceptable salt thereof.

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