Skin care composition as well as preparation method and application thereof

Abstract

The embodiment of the invention provides a skin care composition as well as a preparation method and application thereof, the skin care composition comprises alkyl gallate, tryptophan and mannitol, and the concentration of the alkyl gallate is 0.1 mu mol / L-4 mmol / L. The skin care composition can be prepared into skin care compositions in various dosage forms, is used for skin care and repair, has the effects of sunscreen, anti-inflammation, wrinkle removal and the like, and is simple and convenient to prepare, safe, high in yield and suitable for industrial production.

Description

Technical Field

[0001] The embodiments of the present application belong to the technical field of skin care, and particularly relate to a skin care composition, a preparation method thereof, and an application thereof. Background Art

[0002] Research shows that various factors, including external factors such as sunlight exposure, allergies, soot and dust, and internal factors such as genetic, cell aging caused by age growth, slowed metabolism, and weakened immune system, can all lead to skin aging. Among them, ultraviolet rays and inflammatory cytokines are the main external factors that cause damage or apoptosis of various skin cells, such as keratinocytes, fibroblasts, and macrophages. The apoptosis of these cells will cause the epidermis of the skin to become thinner and the content of collagen and elastin to decrease, thereby generating wrinkles or triggering skin inflammation. Therefore, there is a need to provide a skin care composition that can simultaneously reduce ultraviolet damage, promote skin cell proliferation, and reduce inflammatory responses, so as to restore or maintain the strength and elasticity of the skin, and delay the rate of skin relaxation and aging. Summary of the Invention

[0003] In view of this, the embodiments of the present application provide a skin care composition, a preparation method thereof, and an application thereof. The skin care composition includes alkyl gallate, tryptophan, and mannitol. Among them, tryptophan can improve the survival rate of skin cells under ultraviolet irradiation, mannitol can inhibit the inflammatory response of monocytes in the skin, and alkyl gallate itself can promote the growth of skin cells and secrete collagen and elastin. At the same time, it can also cooperate with tryptophan and mannitol to improve their anti-ultraviolet activity and anti-inflammatory activity. Applying this skin care composition to the skin surface can reduce ultraviolet damage, maintain or restore the strength and elasticity of the skin, prevent skin inflammation, and delay the rate of skin relaxation and aging.

[0004] In a first aspect, the embodiments of the present application provide a skin care composition, including alkyl gallate, tryptophan, and mannitol. The concentration of alkyl gallate in the skin care composition is 0.1 μmol / L - 4 mmol / L.

[0005] In the embodiments of the present application, the concentration of tryptophan in the skin care composition is 0.1 mmol / L - 1 mol / L, and the concentration of mannitol in the skin care composition is 0.1 mmol / L - 1 mol / L.

[0006] In the embodiments of the present application, in the skin care composition, the molar ratio of alkyl gallate to tryptophan is 1:(5 - 100); the molar ratio of alkyl gallate to mannitol is 1:(50 - 2000).

[0007] In the embodiments of the present application, the alkyl gallate includes one or more of ethyl gallate, propyl gallate, octyl gallate, and dodecyl gallate.

[0008] In the embodiments of the present application, the skin care composition further includes other active ingredients and / or adjuvants.

[0009] In the embodiments of the present application, the other active ingredients include one or more of a moisturizer, a whitening agent, an oil control agent, a repair agent, and an anti-aging agent; the adjuvants include one or more of an emulsifier, a buffer, a pH regulator, a thickening agent, a penetration enhancer, a preservative, and a fragrance.

[0010] In the embodiments of the present application, the dosage form of the skin care composition includes one or more of an aqueous solution, an oil, a mud, an emulsion, an ointment, a cream, a gel, a foam, and an aerosol.

[0011] In a second aspect, an embodiment of the present application provides a method for preparing a skin care composition, including: dissolving an alkyl gallate, tryptophan, and mannitol in a solvent to obtain the skin care composition, wherein the concentration of the alkyl gallate in the skin care composition is 0.1 μmol / L - 4 mmol / L.

[0012] In a third aspect, an embodiment of the present application further provides an application of the skin care composition provided in the first aspect of the present application or the skin care composition provided in the third aspect of the present application in the preparation of a product for skin care and improving skin conditions.

[0013] In the embodiments of the present application, the skin conditions include sun damage, wrinkles, inflammation, skin redness, skin dullness, and damaged skin barrier.

[0014] The skin care composition provided by the embodiment of the present application can simultaneously reduce ultraviolet damage, promote skin cell proliferation, and reduce inflammatory reactions, and has obvious effects on restoring or maintaining the strength and elasticity of the skin. Description of the Drawings

[0015] In order to more clearly illustrate the technical solutions in the embodiments of the present application or the prior art, the following will briefly introduce the drawings required for use in the description of the embodiments or the prior art. The specific embodiments described herein are only used to explain the present application and are not used to limit the present application.

[0016] Figure 1 It is the cell survival rate of keratinocytes after ultraviolet irradiation detected by the CCK-8 method, wherein Figure 1 (a) is the cell survival rate of keratinocytes in tryptophan culture medium, (b) is the cell survival rate of keratinocytes in histidine culture medium, (c) is the cell survival rate of keratinocytes in mannitol culture medium, and (d) is the cell survival rate of keratinocytes in tryptophan culture medium and MIX culture medium;

[0017] Figure 2 It is the flow cytometry detection result of keratinocytes after ultraviolet irradiation, whereinFigure 2 (a) shows the cell viability distribution of keratinocytes in the control group, (b) shows the cell viability distribution of keratinocytes in tryptophan culture medium, (c) shows the cell viability distribution of keratinocytes in histidine culture medium, and (d) shows the cell viability distribution of keratinocytes in mannitol culture medium;

[0018] Figure 3 represents the cell survival rates of keratinocytes and fibroblasts after ultraviolet irradiation detected by the CCK-8 method. Among them, Figure 3 (a) shows the cell survival rate of keratinocytes after 5 minutes of ultraviolet irradiation, (b) shows the cell survival rate of keratinocytes after 10 minutes of ultraviolet irradiation, (c) shows the cell survival rate of fibroblasts after 5 minutes of ultraviolet irradiation, and (d) shows the cell survival rate of fibroblasts after 10 minutes of ultraviolet irradiation;

[0019] Figure 4 represents the flow cytometry detection results of keratinocytes after ultraviolet irradiation. Among them, Figure 4 (a) shows the cell viability distribution in PBS culture medium containing different concentrations of propyl gallate, (b) shows the cell viability distribution in tryptophan culture medium containing different concentrations of propyl gallate, and (c) shows the cell viability distribution in MIX2 culture medium containing different concentrations of propyl gallate;

[0020] Figure 5 represents the flow cytometry detection results of fibroblasts after ultraviolet irradiation. Among them, Figure 5 (a) shows the cell viability distribution in PBS culture medium containing different concentrations of propyl gallate, (b) shows the cell viability distribution in tryptophan culture medium containing different concentrations of propyl gallate, and (c) shows the cell viability distribution in MIX2 culture medium containing different concentrations of propyl gallate;

[0021] Figure 6 represents the cell proliferation rates of keratinocytes and fibroblasts. Among them, Figure 6 (a) shows the cell proliferation rate of keratinocytes in MIX culture medium containing different concentrations of propyl gallate, and (b) shows the cell proliferation rate of fibroblasts in MIX culture medium containing different concentrations of propyl gallate;

[0022] Figure 7 represents the secretion amounts of collagen and elastin of fibroblasts. Among them, Figure 7 (a) shows the secretion amount of type I collagen, and (b) shows the secretion amount of elastin;

[0023] Figure 8 represents the CD54 expression of macrophages in different culture media, Figure 8(a) shows the CD54 expression of macrophages in PBS+PG culture medium, (b) shows the CD54 expression of macrophages in TRP+PG culture medium, (c) shows the CD54 expression of macrophages in HIS+PG culture medium, (d) shows the CD54 expression of macrophages in MNT+PG culture medium, (e) shows the CD54 expression of macrophages in MIX2+PG culture medium;

[0024] Figure 9 is the histogram of the CD54 expression rate of macrophages in different culture media;

[0025] Figure 10 is the CD86 expression of macrophages in different culture media, Figure 10 (a) shows the CD86 expression of macrophages in PBS+PG culture medium, (b) shows the CD86 expression of macrophages in MIX1+PG culture medium, (c) shows the CD86 expression of macrophages in MIX2+PG culture medium, (d) shows the CD86 expression of macrophages in MIX3+PG culture medium,

[0026] Figure 11 is the histogram of the CD86 expression rate of macrophages in different culture media;

[0027] Figure 12 is the comparison diagram of the skin cream provided by an embodiment of the present application for skin tests, where, Figure 12 (a) shows the ear posterior skin image of a 62-year-old Asian male, (b) shows the ear posterior skin image of the 62-year-old Asian male after using the skin cream for two months;

[0028] Figure 13 is the comparison diagram of the skin cream provided by an embodiment of the present application for skin tests, where, Figure 12 (a) shows the back skin image of a 61-year-old Asian female after being exposed to the sun for 4 hours without any measures, (b) shows the back skin image of the 61-year-old Asian female after being exposed to the sun for 4 hours and applying the skin cream;

[0029] Figure 14 is the comparison diagram of the skin cream provided by an embodiment of the present application for skin tests, where, Figure 14 (a) shows the back skin image of a 61-year-old Asian female after being exposed to the sun for 4 hours without any measures, (b) shows the back skin image of the 61-year-old Asian female 12 hours after applying the skin cream after sun exposure;

[0030] Figure 15 is the comparison diagram of the skin cream provided by an embodiment of the present application for skin tests, Figure 15Among them, (a) is an image of allergic dermatitis of the facial skin of an Asian female aged 28, (b) is an image of the facial skin of the 28-year-old Asian female 4 hours after the first application of the skin care cream, (c) is an image of the facial skin of the 28-year-old Asian female 4 hours after the second additional application of the skin care cream, and (d) is an image of the facial skin of the 28-year-old Asian female 4 hours after the third additional application of the skin care cream. Detailed implementation manner

[0031] The technical solutions in the embodiments of the present invention will be clearly and completely described below. Obviously, the described embodiments are only a part of the embodiments of the present invention, rather than all the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those of ordinary skill in the art without creative efforts shall fall within the protection scope of the present invention.

[0032] Unless otherwise specified, the raw materials used in the embodiments of the present invention are all commercially available products.

[0033] Keratinocytes (HaCaT): Belonging to the main cells of the epidermis layer, accounting for more than 90% of epidermal cells, they can form a skin barrier, produce keratin to enhance the mechanical strength of the skin, and at the same time participate in wound repair and the transmission of immune signals.

[0034] Fibroblasts (HFF-1): Belonging to the core cells of the dermis layer, they can synthesize collagen, elastic fibers, and hyaluronic acid, maintain the skin structure and elasticity, and participate in wound healing and inflammation regulation.

[0035] Macrophages: Belonging to the immune mononuclear-macrophage system, they are differentiated from bone marrow hematopoietic stem cells. When the skin is infected or damaged, they migrate to the inflammatory or damaged site of the skin through single blood cells. The macrophages exemplified in this application are differentiated from THP-1 cells.

[0036] CD54: Also known as intercellular adhesion molecule-1 or ICAM-1, belonging to the immunoglobulin superfamily, it is a transmembrane glycoprotein that can help immune cells migrate and aggregate. CD54 is expressed at a low level in the resting state and is upregulated by inflammatory factors. At the site of infection or inflammation, endothelial cells highly express CD54 to recruit leukocytes to exude from blood vessels, and its overexpression may lead to out-of-control inflammation.

[0037] CD86: Also known as B7-2, belonging to the B7 family, it is a transmembrane co-stimulatory molecule that is expressed at a low level in the resting state and can inhibit the over-activation of T cells to maintain immune tolerance. Its overexpression may lead to out-of-control inflammation.

[0038] The skin is generally divided into three layers, including the epidermis, dermis, and subcutaneous tissue. The epidermis is the outer protective layer, mainly containing keratinocytes, melanocytes, Langerhans cells, and Merkel cells; the dermis is the middle layer composed of the papillary layer and reticular layer, mainly containing fibroblasts, macrophages, and mast cells, which are embedded with collagen and elastic fibers (connective tissue) to jointly maintain the structure and elasticity of the skin; the subcutaneous tissue is the deepest layer, mainly including adipocytes with functions such as storing fat, regulating temperature, and absorbing impact force.

[0039] Due to aging and numerous external factors, skin tissue changes. First, the most common exogenous factor is the cumulative damage caused by ultraviolet (UV) irradiation. Studies have shown that 80% of skin laxity and wrinkles are due to apoptosis of keratinocytes and damaged barriers caused by photoaging; second, the main endogenous factor is natural aging, which leads to a decline in fibroblast activity, resulting in weakened cell division ability and reduced gene expression of collagen synthesis, manifested as reduced skin elasticity or even collapse; in addition, in the face of a complex external environment, skin allergies such as redness, itching, desquamation, and blisters often occur. This is an abnormal immune response of the body to external stimuli, which is caused by immune cells such as macrophages leading the initiation, amplification, and resolution of allergic reactions by releasing specific inflammatory factors. CD54 and CD86 are key molecules for immune cell adhesion and co-stimulation, and these two molecules are significantly upregulated during inflammation or immune activation. In the field, the anti-inflammatory effects of skin care products or drugs are often reflected based on their expression changes. Therefore, the main directions for developing skin care products to delay skin aging should be to resist UV damage, maintain skin cell activity, and control inflammatory reactions.

[0040] An embodiment of the present application provides a skin care composition, including alkyl gallate, tryptophan, and mannitol, wherein the concentration of alkyl gallate in the skin care composition is 0.1 μmol / L - 4 mmol / L.

[0041] The skin care composition provided by the embodiment of the present application uses alkyl gallate, tryptophan, and mannitol as the main active ingredients. Among them, tryptophan can improve the survival rate of epidermal cells and dermal cells under UV irradiation, mannitol has an inhibitory effect on the release of inflammatory factors by macrophages in the skin, and alkyl gallate itself can promote the proliferation of dermal cells and secrete collagen and elastin. At the same time, alkyl gallate can also enhance the anti-UV activity of tryptophan and synergistically with mannitol further inhibit the expression of inflammatory factors. Applying the skin care composition with these three substances as active ingredients to the skin can effectively reduce UV damage, maintain skin elasticity, and reduce allergic reactions.

[0042] In the embodiments of the present application, the concentration of alkyl gallate in the skin care composition is 0.1 μmol / L - 4 mmol / L. In some embodiments of the present application, the concentration of alkyl gallate in the skin care composition can be, for example, but not limited to, 0.1 μmol / L, 1 μmol / L, 10 μmol / L, 20 μmol / L, 30 μmol / L, 40 μmol / L, 50 μmol / L, 60 μmol / L, 70 μmol / L, 80 μmol / L, 90 μmol / L, 100 μmol / L, 500 μmol / L, 800 μmol / L, 1 mmol / L, 2 mmol / L, 2.5 mmol / L, 3 mmol / L, 4 mmol / L. As the core active ingredient of the skin care composition provided by the present application, alkyl gallate has the functions of promoting the proliferation of dermal cells and secreting collagen to maintain skin elasticity, and reducing the expression of inflammatory factors to inhibit skin inflammatory reactions. Controlling its concentration within the above range can ensure that its content is sufficient to exert better skin care effects, while avoiding adverse reactions caused by excessive amounts. In some specific embodiments of the present application, the concentration of alkyl gallate in the skin care composition is 50 μmol / L - 100 μmol / L.

[0043] In the embodiments of the present application, the structural formula of alkyl gallate is shown in formula (Ⅰ), wherein R is an alkyl group with 2 - 12 carbon atoms. In some embodiments of the present application, the number of carbon atoms in the R group can be 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12.

[0044]

[0045] In some specific embodiments of the present application, the alkyl gallate includes one or more of ethyl gallate (EG), propyl gallate (PG), octyl gallate (OG), and lauryl gallate (LG). Among them, the structure of ethyl gallate is shown in formula (Ⅰ-1), the structure of propyl gallate is shown in formula (Ⅰ-2), the structure of octyl gallate is shown in formula (Ⅰ-3), and the structure of lauryl gallate is shown in formula (Ⅰ-4).

[0046]

[0047] In some embodiments of the present application, the alkyl gallate further includes derivatives of alkyl gallate, and the present application does not limit this derivative, which can be a substance in which alkyl gallate is substituted by groups such as halogen, oxygen, sulfur, nitrogen, glycosyl, and amino acid.

[0048] In the embodiments of the present application, the structure of tryptophan is shown in Formula (II), and the concentration of tryptophan in the skin care composition is 0.1 mmol / L - 1 mol / L. In some embodiments of the present application, the concentration of tryptophan in the skin care composition can be, for example, but not limited to, 0.1 mmol / L, 0.5 mmol / L, 1 mmol / L, 2 mmol / L, 3 mmol / L, 4 mmol / L, 5 mmol / L, 7 mmol / L, 9 mmol / L, 10 mmol / L, 20 mmol / L, 50 mmol / L, 60 mmol / L, 70 mmol / L, 80 mmol / L, 90 mmol / L, 0.1 mol / L, 0.5 mol / L, 0.8 mol / L, 1 mol / L. As the core active ingredient of the skin care composition provided by the present application, tryptophan can reduce the mortality rate of skin cells under ultraviolet irradiation, reduce skin damage caused by ultraviolet rays, and controlling its concentration within the above range can ensure that its content is sufficient to exert a better anti-ultraviolet effect while avoiding adverse reactions caused by excessive amounts. In some specific embodiments of the present application, the concentration of tryptophan in the skin care composition is 1 mmol / L - 4 mmol / L.

[0049]

[0050] In the embodiments of the present application, the structure of mannitol is shown in Formula (III), and the concentration of mannitol in the skin care composition is 0.1 mmol / L - 1 mol / L. In some embodiments of the present application, the concentration of mannitol in the skin care composition can be, for example, but not limited to, 0.1 mmol / L, 1 mmol / L, 2 mmol / L, 5 mmol / L, 10 mmol / L, 15 mmol / L, 20 mmol / L, 25 mmol / L, 30 mmol / L, 50 mmol / L, 60 mmol / L, 0.1 mol / L, 0.12 mol / L, 0.5 mol / L, 0.6 mol / L, 0.7 mol / L, 0.8 mol / L, 1 mol / L. As the core active ingredient of the skin care composition provided by the present application, mannitol helps to reduce the expression of inflammatory factors and inhibit skin inflammatory reactions, enhancing the ability of the skin to resist external stimuli. Controlling its concentration within the above range can ensure that its content is sufficient to exert a better anti-inflammatory effect while avoiding adverse reactions caused by excessive amounts. In some specific embodiments of the present application, the concentration of mannitol in the skin care composition is 15 mmol / L - 60 mmol / L.

[0051]

[0052] In the embodiments of the present application, in the skin care composition, the molar ratio of alkyl gallate to tryptophan is 1:(5 - 100). In some embodiments of the present application, the molar ratio of alkyl gallate to tryptophan can be, for example but not limited to, 1:5, 1:8, 1:10, 1:15, 1:20, 1:25, 1:30, 1:40, 1:50, 1:60, 1:70, 1:80, 1:90, 1:100. Tryptophan itself has anti-ultraviolet activity, specifically manifested as reducing the mortality rate of skin cells under ultraviolet irradiation, thereby preventing sunburn and slowing down skin relaxation. Among them, skin cells include epidermal cells and dermal cells, such as keratinocytes and fibroblasts. On this basis, introducing alkyl gallate can further enhance the protective function of tryptophan on skin cells. Therefore, controlling the molar ratio of alkyl gallate to tryptophan within the above range can ensure that there is a sufficient amount of alkyl gallate to further stimulate the anti-ultraviolet activity of tryptophan, and the two produce a synergistic effect to endow the skin care composition with better protective efficacy, while avoiding adverse effects caused by excessive alkyl gallate. In some specific embodiments of the present application, the molar ratio of alkyl gallate to tryptophan in the skin care composition is 1:(10 - 80).

[0053] In the embodiments of the present application, in the skin care composition, the molar ratio of alkyl gallate to mannitol is 1:(50 - 2000). For example, it can be, but not limited to, 1:50, 1:100, 1:150, 1:200, 1:250, 1:300, 1:350, 1:500, 1:600, 1:800, 1:1000, 1:1200, 1:1400, 1:1600, 1:1800, 1:2000. Both alkyl gallate and mannitol can inhibit the inflammatory reaction caused by the activation of immune monocytes, specifically manifested as reducing the CD54 expression and CD86 expression of monocytes such as macrophages. Controlling the molar ratio of alkyl gallate to mannitol within the above ratio range can ensure that the two play an anti-inflammatory role without interfering with each other, and even produce an effect greater than the superposition of the effects of the two. In some specific embodiments of the present application, the molar ratio of alkyl gallate to mannitol is 1:(150 - 1200).

[0054] In some specific embodiments of the present application, in the skin care composition, the molar ratio of alkyl gallate, tryptophan to mannitol is 1:(10 - 80):(150 - 1200).

[0055] In the embodiments of the present application, the skin care composition further includes a solvent. The solvent is used to disperse, mix and dissolve alkyl gallate, tryptophan and mannitol, and no reaction occurs between the solvent and alkyl gallate, tryptophan, mannitol.

[0056] In the embodiments of the present application, the pH value of the skin care composition is 2-9. For example, it can be, but is not limited to, 2, 3, 4, 4.2, 4.5, 4.7, 4.9, 5.2, 5.5, 5.7, 6, 6.2, 6.5, 6.8, 7, 7.2, 7.5, 7.7, 7.9, 8, 9, etc. In some embodiments of the present application, the pH value of the skin care composition can be 4-8, 5-8, 5-7.9, 5.1-7.9, 6-7.9, 6-7.6, 6.1-7.6, 6.2-7.6, 6.3-7.6, 6.4-7.6, 6.5-7.5, 7-7.4, etc. In some specific embodiments of the present application, the pH value of the skin care composition is 6.5-7.5, which belongs to the neutral range and is more suitable for preservation without affecting the efficacy. In some other specific embodiments of the present application, the pH value of the skin care composition is 4.5-6, which belongs to the weak acidic range and is closer to the natural weak acidic environment of the skin, which is beneficial to maintaining the skin barrier function, enhancing stability, and optimizing the effect of active ingredients.

[0057] In the embodiments of the present application, the skin care composition further includes other active ingredients and / or adjuvants.

[0058] In some embodiments of the present application, the other active ingredients in the skin care composition include, but are not limited to, one or more of a humectant, a whitening agent, an oil control agent, a repair agent, and an anti-aging agent. The humectant can be, for example, but is not limited to, one or more of glycerin, propylene glycol, butylene glycol, glucose, sorbitol, hyaluronic acid, polyglutamic acid, collagen, and vitamins; the whitening agent can be, for example, but is not limited to, one or more of ascorbyl palmitate, niacinamide, tranexamic acid, arbutin, kojic acid, and fruit acids; the oil control agent can be, for example, but is not limited to, one or more of salicylic acid, oligopeptide-1, retinoic acid, and peony essence; the repair agent can be, for example, but is not limited to, one or more of squalane, calendula, astaxanthin, ceramide, and tripeptide-1; the anti-aging agent can be, for example, but is not limited to, one or more of retinol, peptides, hydroxyprogesterone caproate, and epidermal growth factor.

[0059] In some embodiments of the present application, the adjuvants in the skin care composition include, but are not limited to, one or more of an emulsifier, a thickener, a penetration enhancer, a preservative, a fragrance, a buffer, and a pH regulator. The emulsifier can be, for example but not limited to, one or more of cetearyl alcohol, caprylic / capric triglyceride, sodium lauryl sulfate, sodium stearoyl glutamate, quaternary ammonium salt, lecithin, xanthan gum, olive oil, beeswax, silicone emulsifier, and co-emulsifier; the thickener can be, for example but not limited to, one or more of carbomer, xanthan gum, sodium alginate, paraffin, sodium polyacrylate, acrylate copolymer; the penetration enhancer can be, for example but not limited to, one or more of diethylene glycol monoethyl ether, ethanol, and propylene glycol; the preservative can be, for example but not limited to, one or more of parabens, isothiazolinones, organic acids, and polyols; the fragrance can be, for example but not limited to, one or more of phenethyl alcohol, geraniol, limonene, linalool, essential oils, and natural extracts; the buffer can be, for example but not limited to, one or more of phosphate buffer, citric acid-sodium citrate buffer, lactic acid-sodium lactate buffer, and histidine; the pH regulator includes one or more of organic acids, inorganic acids, organic bases, inorganic bases, and buffer solutions, and can be, for example but not limited to, one or more of histidine, lactic acid, phosphoric acid, citric acid, glycolic acid, ascorbic acid, sodium hydroxide, triethanolamine, arginine, aminomethylpropanol, phosphate buffer, citric acid-sodium citrate buffer, and lactic acid-sodium lactate buffer.

[0060] In the embodiments of the present application, the skin care composition includes various common dosage forms, such as, but not limited to, aqueous solution, oil, mud, emulsion, paste, cream, gel, foam, and aerosol. Based on the solubility differences of alkyl gallates, in some embodiments of the present application, the alkyl gallate in the aqueous skin care composition is ethyl gallate, the alkyl gallate in the emulsion or cream skin care composition is propyl gallate, the alkyl gallate in the oil skin care composition is octyl gallate, and the alkyl gallate in the paste skin care composition is dodecyl gallate.

[0061] The skin care composition provided by the embodiments of the present application first utilizes the individual effects of the three core active ingredients. When the concentration of alkyl gallate in the skin care composition is 0.1 μmol / L - 4 mmol / L, it can ensure its anti-inflammatory activity and promote skin cell growth function without irritating the skin. Tryptophan can improve the survival rate of epidermal cells and dermal cells under ultraviolet irradiation, and mannitol can inhibit the release of inflammatory factors by macrophages in the skin. Thus, the skin care composition has the functions of skin maintenance and repair. Further, through the specific ratio between the components, the synergistic effect between the two is maximized, and the protective function of each component on the skin is stimulated to a greater extent, so that the skin care composition provided by the embodiments of the present application has excellent effects in aspects such as sunscreen, anti-inflammation, and anti-wrinkle.

[0062] The embodiments of the present application provide a preparation method of a skin care composition, including:

[0063] Dissolve an alkyl gallate, tryptophan and mannitol in a solvent to obtain a skin care composition, wherein the concentration of the alkyl gallate in the skin care composition is 0.1 μmol / L - 4 mmol / L.

[0064] In the embodiments of the present application, the solvent includes but is not limited to water and an acid-base balance solution. The acid-base balance solution can be, for example, but is not limited to one or more of phosphate buffer solution (PBS), citric acid-sodium citrate buffer solution, and lactic acid-sodium lactate buffer solution.

[0065] In the embodiments of the present application, the skin care composition in any of the above embodiments can be prepared by this preparation method. The preparation method of the skin care composition provided by the embodiments of the present application has the advantages of wide raw material sources, easy availability, simple operation, high safety, and high product yield, and is suitable for large-scale industrial production.

[0066] The embodiments of the present application also provide an application of the aforementioned skin care composition or the skin care composition prepared by the aforementioned preparation method in products for skin care and improving skin conditions.

[0067] In the embodiments of the present application, the application method of the skin care composition includes applying the skin care composition to the skin surface.

[0068] In some embodiments of the present application, skin care includes but is not limited to soothing, moisturizing, sun protection, anti-inflammatory, anti-allergic, anti-wrinkle and anti-aging.

[0069] In some embodiments of the present application, skin conditions include but are not limited to sun damage, wrinkles, inflammation, skin redness, skin dullness and damaged skin barrier. Improving skin conditions includes but is not limited to post-sun repair, sunburn repair, wrinkle reduction, swelling and inflammation relief, skin brightening and skin barrier repair.

[0070] In some embodiments of the present application, products for skin care and improving skin conditions include anti-wrinkle products, anti-crease products, sun protection products, post-sun repair products, anti-inflammatory products, anti-aging products, products that inhibit the expression of CD54 and CD86, etc.

[0071] In the embodiments of the present application, the skin care composition is applicable to people of all ages, including but not limited to infants, children, teenagers, adults and the elderly.

[0072] The embodiments of the present application also provide a skin care method, including applying the skin care composition in any of the above embodiments to the skin surface.

[0073] The embodiments of the present application also provide a method for improving skin conditions, including applying the skin care composition in any of the above embodiments on the skin surface.

[0074] The effects of the technical solutions of the present application are further described below through specific examples. The following examples are all carried out at room temperature.

[0075] HaCaT (human immortalized epidermal keratinocytes), HFF-1 (human skin fibroblasts), and THP-1 (human monocytes) in the examples of the present application were all purchased from ATCC (Manassas, VA, USA).

[0076] In the examples of the present application, ultraviolet irradiation was used to irradiate keratinocytes HaCaT and fibroblasts HFF-1 to simulate the cell states of human epidermal cells and dermal cells under ultraviolet irradiation, so as to verify the anti-ultraviolet activity of the skin care composition provided by the present application; macrophages formed after the differentiation of THP-1 by PMA were used to simulate the inflammatory reaction of the skin, so as to verify the anti-inflammatory activity of the skin care composition provided by the present application.

[0077] The culture media in the examples of the present application include PBS culture medium with a pH value of 7.2, tryptophan (hereinafter abbreviated as "TRP") culture medium, histidine (hereinafter abbreviated as "HIS") culture medium, mannitol (hereinafter abbreviated as "MNT") culture medium, MIX1 culture medium, MIX2 culture medium, MIX3 culture medium, and the above culture media containing propyl gallate (hereinafter abbreviated as "PG"). Among them, the PBS culture medium is used as a control group. The MIX1 culture medium includes 1 mM tryptophan, 55 mM histidine, and 15 mM mannitol. The MIX2 culture medium includes 2 mM tryptophan, 110 mM histidine, and 30 mM mannitol. The MIX3 culture medium includes 4 mM tryptophan, 220 mM histidine, and 60 mM mannitol. Among them, histidine is used as a pH regulator and buffer, and the solvent of each culture medium is phosphate buffer (disodium hydrogen phosphate + potassium dihydrogen phosphate).

[0078] The CCK-8 method was used in the examples of the present application to detect the cell viability: viability = (OD reading of the sample - OD reading of the blank sample) ÷ (OD reading of the untreated sample - OD reading of the blank sample) × 100%, and the data was expressed as mean ± standard deviation.

[0079] In the examples of the present application, a flow cytometer was used to monitor the cell state, and the counting rule is: the upper left grid represents necrotic cells, the upper right grid represents late apoptotic cells, the lower left grid represents normal living cells, and the lower right grid represents early apoptotic cells.

[0080] This application example reflects the anti-inflammatory activity of a skin care composition by detecting the CD54 expression and CD86 expression of macrophages. CD54 and CD86 are key molecules for immune cell adhesion and co-stimulation. These two molecules are significantly upregulated during inflammation or immune activation. The anti-inflammatory effect of the skin care composition can be reflected by their expression changes. If the expressions of CD54 and CD86 can be significantly reduced, it indicates that it has anti-inflammatory activity.

[0081] Example 1: Protective effects of tryptophan, histidine, and mannitol on keratinocyte HaCaT under ultraviolet irradiation

[0082] HaCaT cells were cultured and adhered for 24 hours (h), then the cell culture medium was removed. The cells were respectively stored in equal amounts in PBS culture solution, different concentrations of tryptophan culture solution (TRP concentrations were 1 mM, 2 mM, 10 mM, 20 mM), different concentrations of histidine culture solution (HIS concentrations were 28 mM, 55 mM, 110 mM, 129 mM), different concentrations of mannitol culture solution (MNT concentrations were 3 mM, 30 mM, 60 mM, 120 mM), MIX1 culture solution, and MIX2 culture solution. After being irradiated with ultraviolet rays at 30 mJ / cm 2 for 1.5 minutes (min) to 15 minutes, the cell culture medium was replaced and the cells were cultured for another 24 hours. The CCK-8 method was used to detect the survival rate of HaCaT cells, and the results are as Figure 1 shown:

[0083] Figure 1 In (a), it is the cell survival rate of keratinocytes in different concentrations of tryptophan culture solution. Compared with the control group PBS culture solution, the keratinocytes in the tryptophan culture solution had significantly higher survival rates, indicating that 1 mM tryptophan culture solution could protect keratinocytes under ultraviolet irradiation, and the tryptophan culture solutions with 2 mM and above showed almost the same ultraviolet protection activity. By two-way analysis of variance (n = 3): the p-value for the comparison between 1 mM tryptophan culture solution and PBS culture solution was less than 0.0001, the p-value for the comparison between 2 mM tryptophan culture solution and 1 mM tryptophan culture solution was less than 0.0001, the p-value for the comparison between 10 mM tryptophan culture solution and 2 mM tryptophan culture solution was 0.9652, and the p-value for the comparison between 20 mM tryptophan culture solution and 10 mM tryptophan culture solution was 0.5031;

[0084] Figure 1(b) and (c) in it are the cell survival rates of keratinocytes in culture media with different concentrations of histidine and mannitol respectively. From the data, it can be seen that under ultraviolet irradiation, neither histidine nor mannitol has a protective effect on keratinocytes. Through two-way analysis of variance (n = 3): the p-value for comparison between the 28 mM histidine culture medium and the PBS culture medium in (b) is 0.1123, and the p-value for comparison between the 129 mM histidine culture medium and the PBS culture medium is 0.1064; the p-value for comparison between the 3 mM mannitol culture medium and the PBS culture medium in (c) is 0.7227, and the p-value for comparison between the 120 mM histidine culture medium and the PBS culture medium is 0.5868;

[0085] Figure 1 (d) in it is the cell survival rate of keratinocytes in culture media with different concentrations of tryptophan, MIX1 culture medium, and MIX2 culture medium. The data shows that the survival rates of keratinocytes in MIX1 culture medium and 1 mM tryptophan culture medium are the same, and the survival rates of keratinocytes in MIX2 culture medium and 2 mM tryptophan culture medium are the same, indicating that among the three substances, only tryptophan has a protective effect on keratinocytes, and when the three coexist, histidine and mannitol have no effect on the ultraviolet protection activity of tryptophan. Through two-way analysis of variance (n = 3): the p-values for comparison between the 1 mM tryptophan culture medium and MIX1 culture medium and the PBS culture medium in (d) are both less than 0.0001, the p-values for comparison between the 2 mM tryptophan culture medium and MIX2 culture medium and the PBS culture medium are both less than 0.0001, the p-value for comparison between the 1 mM tryptophan culture medium and MIX1 culture medium is less than 0.1263, and the p-value for comparison between the 2 mM tryptophan culture medium and MIX2 culture medium is 0.5691.

[0086] It can be known from Figure 1 that tryptophan has ultraviolet protection activity on keratinocytes and can effectively reduce the mortality rate of keratinocytes under short-term or long-term ultraviolet irradiation. When tryptophan, histidine, and mannitol are used together as active ingredients of a skin care composition, the ultraviolet protection activity of tryptophan is not affected.

[0087] Furthermore, the state distribution of keratinocytes was monitored by flow cytometry through 7-AAD and Annexin-V staining to verify the above conclusion: Keratinocytes were preserved in PBS culture medium, culture media with different concentrations of tryptophan (TRP concentration was 2 mM, 10 mM), culture media with different concentrations of histidine (HIS concentration was 55 mM, 100 mM), and culture media with different concentrations of mannitol (MNT concentration was 30 mM, 60 mM). After being irradiated with 30 mJ / cm 2 ultraviolet light for 10 minutes, and then switched back to the cell culture medium for 24 hours, the active distribution state of keratinocytes in each group was monitored by flow cytometry. The results are as Figure 2 shown, among which, Figure 2(a) shows the cell viability distribution of keratinocytes in the control group. "Untreated control" refers to keratinocytes preserved in PBS solution without ultraviolet irradiation. (b) shows the cell viability distribution of keratinocytes in culture media with different concentrations of tryptophan. (c) shows the cell viability distribution of keratinocytes in culture media with different concentrations of histidine. (d) shows the cell viability distribution of keratinocytes in culture media with different concentrations of mannitol. It can be seen from Figure 2 that: for keratinocytes preserved in PBS culture medium, the number of normal living cells is 88.56% without ultraviolet treatment, and it drops to 36.12% after 10 minutes of ultraviolet irradiation; the cell viability distributions of keratinocytes preserved in histidine culture medium and mannitol culture medium are almost the same as those in PBS culture medium, indicating that regardless of the concentrations of histidine and mannitol, they cannot prevent the apoptosis process of keratinocytes under ultraviolet irradiation; when keratinocytes are preserved in 2 mM tryptophan culture medium and 10 mM tryptophan culture medium, the number of normal living cells increases to 70.31% and 93.53% respectively, proving that tryptophan has a significant ultraviolet protection effect on keratinocytes, and its protection intensity is positively correlated with the concentration of tryptophan within a certain range.

[0088] Therefore, it can be jointly verified by Figure 1 and Figure 2 that tryptophan plays a strong protective role on keratinocytes under ultraviolet irradiation, can effectively reduce cell apoptosis caused by ultraviolet irradiation, and this protective effect is not affected by histidine (pH regulator or buffer) and mannitol.

[0089] Example 2: Propyl gallate can enhance the protective effect of tryptophan on skin cells under ultraviolet irradiation

[0090] HaCaT cells and HFF-1 cells were cultured and adhered for 24 hours, then the cell culture medium was removed, and they were respectively and equally preserved in PBS culture medium, MIX1 culture medium, MIX2 culture medium and MIX3 culture medium. Different concentrations (0 to 100 μM) of propyl gallate were added, and after 5 minutes or 10 minutes of ultraviolet irradiation at 30 mJ / cm 2 , the cells were then changed back to the cell culture medium and continued to be cultured for 24 hours. The CCK-8 method was used to detect the cell survival rate, and the results are as Figure 3 shown.

[0091] Figure 3The cell survival rates of HaCaT cells after 5 minutes and 10 minutes of ultraviolet irradiation in different MIX culture media containing different concentrations of propyl gallate are shown in (a) and (b) respectively. It can be seen from (a) and (b) that the survival rate of HaCaT cells increases with the increase in the concentration of MIX culture medium. After adding propyl gallate to each MIX culture medium, the cell survival rate also increases to a certain extent, indicating that adding propyl gallate to the MIX culture medium can further enhance the protective effect on keratinocytes, and the intensities of the ultraviolet protection enhancement by propyl gallate at concentrations of 50 μM and 100 μM are almost the same. Through two-way ANOVA (n = 3): In (a), the p-value for the comparison between the MIX culture medium containing 50 μM propyl gallate and the MIX culture medium without propyl gallate is 0.0003, the p-value for the comparison between the MIX culture medium containing 100 μM propyl gallate and the MIX culture medium without propyl gallate is 0.0242, and the p-value for the comparison between the MIX culture medium containing 100 μM propyl gallate and the MIX culture medium containing 50 μM propyl gallate is 0.931; in (b), the p-value for the comparison between the MIX culture medium containing 50 μM propyl gallate and the MIX culture medium without propyl gallate is 0.0002, the p-value for the comparison between the MIX culture medium containing 100 μM propyl gallate and the MIX culture medium without propyl gallate is 0.001, and the p-value for the comparison between the MIX culture medium containing 100 μM propyl gallate and the MIX culture medium containing 50 μM propyl gallate is 0.5867;

[0092] Figure 3In (c) and (d), they are the cell survival rates of HFF-1 cells stored in different MIX culture media containing different concentrations of propyl gallate after being irradiated with ultraviolet light for 5 minutes and 10 minutes respectively. From (c) and (d), it can be seen that the survival rate of HFF-1 cells increases with the increase in the concentration of the MIX mixture. After adding propyl gallate to each MIX culture medium, the cell survival rate also increases to a certain extent. Especially for the protective enhancement effect of MIX1 culture medium, it is particularly obvious. The intensity of the ultraviolet protection enhancement effect of 50 μM and 100 μM concentrations of propyl gallate is almost the same, indicating that adding propyl gallate to the MIX culture medium can further enhance the protective effect on fibroblasts, and when PG:TRP = 1:20, this protective enhancement effect is better. Through two-way analysis of variance (n = 3): In (c), the p-value of the comparison between the MIX culture medium containing 50 μM propyl gallate and the MIX culture medium without propyl gallate is less than 0.0001, the p-value of the comparison between the MIX culture medium containing 100 μM propyl gallate and the MIX culture medium without propyl gallate is less than 0.0001, and the p-value of the comparison between the MIX culture medium containing 100 μM propyl gallate and the MIX culture medium containing 50 μM propyl gallate is 0.9719; In (d), the p-value of the comparison between the MIX culture medium containing 50 μM propyl gallate and the MIX culture medium without propyl gallate is less than 0.0001, the p-value of the comparison between the MIX culture medium containing 100 μM propyl gallate and the MIX culture medium without propyl gallate is less than 0.0001, and the p-value of the comparison between the MIX culture medium containing 100 μM propyl gallate and the MIX culture medium containing 50 μM propyl gallate is 0.0299;

[0093] Furthermore, HaCaT cells were stored in PBS culture medium, 2 mM tryptophan culture medium, MIX2 culture medium, and the above-mentioned culture media containing 100 μM propyl gallate. After being irradiated with 30 mJ / cm 2 ultraviolet light for 10 minutes and then switched back to the cell culture medium for 24 hours, the active distribution status of each group of keratinocytes was measured by flow cytometry using 7-AAD and Annexin-V staining. The results are as Figure 4 shown. As Figure 4 can be seen, the number of normal living cells of keratinocytes in PBS culture medium is 51.53%, and the number of normal living cells in PBS culture medium containing propyl gallate rises to 63.81%; compared with tryptophan culture medium, the number of normal living cells in tryptophan culture medium containing propyl gallate is 15.81% more; compared with MIX2 culture medium without propyl gallate, the number of normal living cells increases by 15.38% after adding propyl gallate.

[0094] HFF-1 cells were treated in the same way, and the activity distribution of fibroblasts in each culture medium was measured by flow cytometry. The results are as Figure 5 shown. As Figure 5 can be seen, the number of normal living cells of fibroblasts in PBS culture medium was 63.0%, and the number of normal living cells in PBS culture medium containing propyl gallate increased to 71.7%; compared with tryptophan culture medium, the number of normal living cells in tryptophan culture medium containing propyl gallate was 14.8% more; compared with MIX2 culture medium without propyl gallate, the number of normal living cells increased by 14.9% after adding propyl gallate.

[0095] This further confirmed that propyl gallate can increase the number of normal living cells of keratinocytes and fibroblasts, tryptophan helps to maintain the activity of keratinocytes and fibroblasts under ultraviolet irradiation. On this basis, propyl gallate can synergistically with tryptophan to further inhibit the apoptosis of the two cells under ultraviolet irradiation, and histidine and mannitol have no effect on the synergistic anti-ultraviolet effect of tryptophan and propyl gallate.

[0096] Therefore, from Figure 3 , Figure 4 and Figure 5 joint verification shows that propyl gallate can enhance the protective effect of tryptophan on skin cells, including epidermal cells and dermal cells, under ultraviolet irradiation, and can effectively reduce cell apoptosis caused by ultraviolet irradiation, and the synergistic protective effect of the two is not affected by histidine and mannitol.

[0097] Example 3: Propyl gallate has the function of promoting the proliferation of skin cells

[0098] HaCaT cells and HFF-1 cells were cultured and adhered for 24 hours, the cell culture medium was removed, and they were respectively stored in equal amounts in the mixture of different culture media (PBS culture medium, MIX1 culture medium, MIX2 culture medium, MIX3 culture medium) and cell culture medium. The volume ratio of the culture medium to the cell culture medium in the mixture was 20:80, and then different concentrations (0 to 100 μM) of propyl gallate (PG) were added, and the cell culture was continued for 24 hours. The CCK-8 method was used to detect the proliferation rate of cells. The results are as Figure 6 shown.

[0099] Figure 6 In (a) is the cell proliferation rate of keratinocytes in different culture media. As Figure 6As can be seen from (a), adding propyl gallate to each MIX culture medium can promote the proliferation of keratinocytes, and its proliferation-promoting effect is positively correlated with the concentration of propyl gallate. In (a), the p-value for the comparison between the three groups of MIX culture media and the PBS culture medium was 0.0003 (one-way ANOVA, n = 3), and the p-values for the comparison between the MIX culture medium containing 50 μM propyl gallate and the MIX culture medium containing 100 μM propyl gallate and the PBS culture medium were less than 0.0001 (two-way ANOVA, n = 3). The p-value for the comparison between the MIX culture medium containing 100 μM propyl gallate and the MIX culture medium containing 50 μM propyl gallate was less than 0.0001 (two-way ANOVA, n = 3).

[0100] Figure 6 (b) in shows the cell proliferation rate of fibroblasts in different culture media. From Figure 6 As can be seen from (b) in, adding propyl gallate to each MIX culture medium can promote the proliferation of fibroblasts. In (b), the p-value for the comparison between the three groups of MIX culture media and the PBS culture medium was 0.0006 (one-way ANOVA, n = 3), and the p-value for the comparison between the MIX culture medium containing 100 μM propyl gallate and the MIX culture medium was less than 0.0001 (two-way ANOVA, n = 3).

[0101] Therefore, from Figure 6 it can be seen that propyl gallate has the function of promoting the proliferation of skin cells, including promoting the growth and proliferation of keratinocytes and fibroblasts.

[0102] Example 4: Propyl gallate has the function of promoting fibroblasts to secrete collagen and elastin

[0103] HFF-1 cells were cultured and adhered for 24 hours, the cell culture medium was removed, and they were respectively stored in equal amounts in the mixtures of different culture media (PBS culture medium, MIX1 culture medium, MIX2 culture medium, MIX3 culture medium) and the cell culture medium. The volume ratio of the culture medium to the cell culture medium in the mixture was 20:80. Then, propyl gallate with a concentration of 100 μM was added to half of each type of mixture, and the cell culture was continued for 48 hours. An enzyme-linked immunosorbent assay kit was used to measure the content of type I collagen and elastin in each culture medium. The data was expressed as the mean ± standard deviation, and the results are as Figure 7 shown, where "Vehicle" represents the culture medium without PG.

[0104] Figure 7In (a), the content of type I collagen in different culture media is shown. From (a), it can be seen that the secretion amount of type I collagen by fibroblasts in PBS culture medium and the three MIX culture media is roughly the same. After adding propyl gallate, the content of type I collagen in each culture medium increases significantly. In (a), the p-value for comparison between the three MIX culture media and the PBS culture medium is 0.2493 (one-way ANOVA, n = 3), and the p-value for comparison between the MIX culture medium containing 100 μM propyl gallate and the MIX culture medium without propyl gallate is less than 0.0001 (two-way ANOVA, n = 3).

[0105] Figure 7 In (b), the content of elastin in different culture media is shown. From (b), it can be seen that the secretion amount of elastin by fibroblasts in PBS culture medium and the three MIX culture media is roughly the same. After adding propyl gallate, the content of elastin in each culture medium increases significantly. In (a), the p-value for comparison between the three MIX culture media and the PBS culture medium is 0.3168 (one-way ANOVA, n = 3), and the p-value for comparison between the MIX culture medium containing 100 μM propyl gallate and the MIX culture medium without propyl gallate is less than 0.0001 (two-way ANOVA, n = 3).

[0106] Therefore, from Figure 7 it can be seen that propyl gallate can promote fibroblasts to secrete collagen and elastin.

[0107] Example 5: Propyl gallate and mannitol can inhibit the expression of CD54 and CD86 in monocytes or macrophages

[0108] Add 200 ng / mL PMA (Phorbol 12-myristate 13-acetate) to the medium of THP-1 monocytes to induce the differentiation of monocytes into macrophages (inflammatory response model) to test the effects of each component in the skin care composition on macrophage differentiation.

[0109] THP-1 cells are induced to differentiate into macrophages by PMA, and at the same time, different culture media (2 mM tryptophan culture medium, 110 mM histidine culture medium, 30 mM mannitol culture medium, MIX1 culture medium, MIX2 culture medium, MIX3 culture medium) are added to the medium. The volume ratio of the culture medium to the cell medium is 20:80. At the same time, 100 μM propyl gallate is added to the mixture of the two, and the cell culture is continued for 48 hours. Use a flow cytometer to measure the expression of the inflammatory response marker molecules CD54 and CD86 in macrophages, and the expression levels are statistically analyzed by flow cytometry histograms. The results are as Figure 8 、 Figure 9 、 Figure 10 andFigure 11 as shown

[0110] Figure 8 shows the CD54 expression of macrophages in different culture media Figure 8 In the figure, "control" represents the image background, the black dashed line represents unstimulated control cells, the red line represents cells stimulated with PMA, the blue line represents cells stimulated with PMA and treated with the culture medium, and the black solid line represents cells stimulated with PMA and treated with the culture medium containing propyl gallate Figure 8 In (a), it reflects the CD54 expression of macrophages in PBS+PG culture medium; in (b), it reflects the CD54 expression of macrophages in TRP+PG culture medium; in (c), it reflects the CD54 expression of macrophages in HIS+PG culture medium; in (d), it reflects the CD54 expression of macrophages in MNT+PG culture medium; in (e), it reflects the CD54 expression of macrophages in MIX2+PG culture medium

[0111] Figure 9 is a histogram of the CD54 expression rate of macrophages in different culture media, which is a statistical chart based on Figure 8 and more intuitively reflects the Figure 8 detection results Figure 9 In the figure, "Vehicle" represents the culture medium without PG. The p-value for the comparison between tryptophan culture medium and PBS culture medium is 0.6608 (t-test, n = 4); the p-value for the comparison between histidine culture medium and PBS culture medium is 0.2117 (t-test, n = 4); the p-value for the comparison between mannitol culture medium and PBS culture medium is less than 0.0001 (t-test, n = 4); the p-value for the comparison between MIX2 culture medium and PBS culture medium is less than 0.0001 (t-test, n = 4); the p-value for the comparison between MIX2 culture medium and mannitol culture medium is less than 0.0001; the p-value for the comparison between each culture medium containing PG and the corresponding culture medium without PG is less than 0.0001

[0112] by Figure 8 and Figure 9It can be seen that, on the one hand, the CD54 expression of macrophages is almost the same in PBS culture medium, TRP culture medium and HIS culture medium, and the CD54 expression is significantly decreased in MNT culture medium and MIX2 culture medium; on the other hand, compared with the culture media without PG, the CD54 expression of macrophages in the corresponding culture media added with PG is greatly decreased, and even the value of the latter is less than one-half of the former; on the third hand, the CD54 expression is inhibited because MNT is contained in MIX2 culture medium, and after adding PG on this basis, PG synergizes with MNT to further decrease the CD54 expression, and its value is the lowest among all experimental groups. It can be seen from this that propyl gallate can synergize with mannitol to jointly decrease the CD54 expression of monocytes (such as macrophages), that is, it can effectively inhibit the inflammatory reaction of the skin.

[0113] Figure 10 is the CD86 expression of macrophages in different culture media, where the meanings of the black dotted line, red line, blue line and black solid line are the same as Figure 8 the same. Figure 10 in which (a) reflects the CD86 expression of macrophages in PBS+PG culture medium, (b) reflects the CD86 expression of macrophages in MIX1+PG culture medium, (c) reflects the CD86 expression of macrophages in MIX2+PG culture medium, and (d) reflects the CD86 expression of macrophages in MIX3+PG culture medium.

[0114] Figure 11 is the histogram of the CD86 expression rate of macrophages in different culture media, which is a statistical chart based on Figure 10 and more intuitively reflects the Figure 10 detection results, Figure 11 in which, "Vehicle" represents the culture medium without PG, the p values of the comparison between the three groups of MIX culture media and PBS culture medium are all less than 0.0001 (two-way ANOVA, n = 4), and the p values of the comparison between the culture media containing PG and the corresponding culture media without PG are less than 0.0001 (two-way ANOVA, n = 4).

[0115] From Figure 10 and Figure 11It can be seen that the CD86 expression of macrophages in the three groups of MIX culture media is lower than that in the PBS culture media, and the numerical relationship is PBS > MIX1 > MIX2 > MIX3, indicating that the inhibitory degree of CD86 expression is positively correlated with the concentration of mannitol in the culture media. On the other hand, compared with the culture media without propyl gallate, the CD86 expression rate in the corresponding culture media containing propyl gallate is significantly reduced, indicating that propyl gallate can also inhibit the CD86 expression of macrophages. Based on this, after adding propyl gallate to the MIX3 culture media with the highest mannitol concentration, its CD86 expression rate is the lowest among all experimental groups. It can be seen that propyl gallate can cooperate with mannitol to jointly reduce the CD86 expression of monocytes (such as macrophages).

[0116] Therefore, from Figures 8 - 11 it can be seen that both propyl gallate and mannitol can inhibit the CD54 expression and CD86 expression of monocytes or macrophages. Using these two substances as the synergistic active ingredients of the skin care composition can effectively inhibit or treat skin inflammation.

[0117] Example 6: A skin care product with propyl gallate, tryptophan and mannitol as the main active ingredients is used for skin sun protection, wrinkle removal and anti - inflammation

[0118] Using propyl gallate, tryptophan and mannitol (molar ratio 1:40:600) as the main active ingredients, adding histidine, glycerol, cetearyl alcohol, caprylic / capric triglyceride, paraffin, sorbitol, ascorbyl palmitate, butylene glycol, diethylene glycol monoethyl ether, lactic acid, glucose and sodium hydroxide as adjuvants, a neutral (pH 7 - 7.4) skin care cream is prepared.

[0119] Apply this skin care cream to the test subjects for skin testing. The specific content is as follows:

[0120] (1) A 62 - year - old Asian male, the skin behind his ear is as Figure 12 shown in (a). Apply the skin care cream to the wrinkle area behind his ear once a day for two months. After that, the skin behind his ear is as Figure 12 shown in (b). From Figure 12 it can be seen that the skin behind the ear of this 62 - year - old Asian male is loose and has obvious wrinkles. After long - term application of the skin care cream, the wrinkles behind his ear are significantly reduced and the skin elasticity is slightly restored, indicating that the skin care cream provided in this example can effectively fade wrinkles and restore skin elasticity.

[0121] (2) A 61 - year - old Asian female, divide the skin on her back into areas and apply the skin care cream and take no protective measures. After exposing her back to the sun for 4 hours, the skin area without any protective measures is as Figure 13 shown in (a), and the skin area where the skin care cream is applied is as Figure 13 shown in (b). FromFigure 13 It can be seen that after long-term exposure to ultraviolet rays, the skin without any protection measures showed severe redness, swelling and blisters, while the skin with skin cream applied only showed slight redness, which shows that the skin cream provided in this example has a sun protection function and can effectively reduce the damage to the skin caused by ultraviolet rays.

[0122] (3) The same 61-year-old Asian woman developed solar dermatitis on her back after sun exposure as shown in (a) of 14. She applied skin cream once on the dermatitis area. After 12 hours, the affected area became Figure 14 As shown in (b). Figure 14 It can be seen that the redness and swelling on the back of the 61-year-old Asian woman subsided 12 hours after applying the skin cream, indicating that the skin cream provided in this example has after-sun repair function and can effectively treat solar dermatitis.

[0123] (4) A 28-year-old Asian female with allergic dermatitis on her face Figure 15 As shown in (a), additional skin care cream is applied to the dermatitis area at intervals of 4 hours. The skin condition 4 hours after the first application of skin care cream is as follows Figure 15 As shown in (b), the skin condition 4 hours after the second application of skin care cream is as follows Figure 15 As shown in (c), the skin condition 4 hours after the third additional application of skin care cream is as follows Figure 15 As shown in (d). Figure 15 It can be seen that within 12 hours from the first application to the additional application of the skin care cream, the redness and swelling symptoms on the face of the 28-year-old Asian woman gradually subsided until the skin returned to normal, indicating that the skin care cream provided in this example has a repair function and can effectively relieve and treat skin inflammation.

[0124] Therefore, by Figures 12 - 15 It can be seen that the skin care cream provided in Example 6 of the present application has functions such as sun protection, wrinkle removal and anti-inflammatory. When applied to the skin surface, it can effectively reduce wrinkles, reduce UV damage, repair sunburn and treat inflammation.

[0125] In summary, Examples 1 to 6 of the present application implemented simulated skin cell experiments and real skin tests, which jointly proved that propyl gallate, tryptophan and mannitol have protective and synergistic protective functions on skin cells. These three substances are used as the core active ingredients of the skin care composition and can be used for daily skin care and damage repair such as sun protection, wrinkle removal and anti-inflammation.

[0126] The above is a preferred embodiment of the present application, but it should not be understood as limiting the scope of the present application. It should be pointed out that for ordinary technicians in this technical field, several improvements and modifications can be made without departing from the principles of the present application, and these improvements and modifications are also considered to be within the scope of protection of the present application.

Claims

1. A skin care composition, characterized in that, It includes alkyl gallate, tryptophan and mannitol, and the concentration of the alkyl gallate in the skin care composition is 0.1 μmol / L - 4 mmol / L.

2. The skin care composition according to claim 1, wherein In the skin care composition, the concentration of the tryptophan is 0.1 mmol / L - 1 mol / L, and the concentration of the mannitol is 0.1 mmol / L - 1 mol / L.

3. The skin care composition according to claim 1 or 2, characterized in that In the skin care composition, the molar ratio of the alkyl gallate to the tryptophan is 1:(5 - 100); the molar ratio of the alkyl gallate to the mannitol is 1:(50 - 2000).

4. The skin care composition according to any one of claims 1-3, characterized in that, The alkyl gallate includes one or more of ethyl gallate, propyl gallate, octyl gallate and dodecyl gallate.

5. The skin care composition according to any one of claims 1-4, characterized in that, The skin care composition further includes other active ingredients and / or adjuvants.

6. The skin care composition according to claim 5, wherein, The other active ingredients include one or more of a humectant, a whitening agent, an oil-control agent, a repair agent and an anti-aging agent; the adjuvants include one or more of an emulsifier, a buffer, a pH regulator, a thickener, a penetration enhancer, a preservative and an aromatic agent.

7. The skin care composition according to any one of claims 1-6, characterized in that, The dosage form of the skin care composition includes one or more of an aqueous solution, an oil, a mud, an emulsion, a cream, a gel, a foam and an aerosol.

8. A method for preparing a skin care composition, characterized in that, It includes: Dissolve alkyl gallate, tryptophan and mannitol in a solvent to obtain a skin care composition, wherein the concentration of the alkyl gallate in the skin care composition is 0.1 μmol / L - 4 mmol / L.

9. Use of the skin care composition according to any one of claims 1 - 7 or the skin care composition prepared by the preparation method of the skin care composition according to claim 8 in the preparation of a product for skin care and improving skin conditions.

10. The use according to claim 9, wherein the skin conditions include one or more of sun damage, wrinkles, inflammation, skin redness, skin dullness and impaired skin barrier.

Images