A kit for resisting skin aging

Abstract

The present invention relates to an anti-skin aging kit, belonging to the field of medical cosmetology technology. The kit provided by the present invention comprises a combination kit of a cocrystal of nicotinamide and a liquid comprising oligopeptides and the like. The kit provided by the present invention can eliminate, alleviate and / or improve the aging phenomenon of the skin, such as dryness, roughness, pigmentation, fine lines and the like.

Description

Technical Field

[0001] The present invention belongs to the field of medical cosmetology technology, and specifically relates to an anti-skin aging kit, and more specifically relates to a combination kit comprising a cocrystal of niacinamide and a liquid comprising oligopeptides and the like; the kit can be used to eliminate, alleviate and / or improve skin aging phenomena, such as dryness, roughness, pigmentation, fine lines and the like, and has anti-aging and other uses. Background Art

[0002] With age and / or changes in the environment, human skin and other organs or tissues will age, showing dry skin, rough skin, seborrheic keratosis, pigmentation and other phenomena or conditions. With the development of society, people's living standards have improved, and the pursuit of beauty has also increased. Therefore, anti-aging, improving skin health, especially eliminating, reducing skin pigmentation, reducing fine lines or preventing the formation of fine lines and other anti-aging methods are the most direct and fast way to pursue beauty, health and anti-aging.

[0003] The external color of the skin, especially the darker and darker color and spots, is related to the content and distribution of melanin; melanin is produced by melanocytes and transported to the stratum corneum of the skin through a series of physiological processes, showing as dark skin color or spots, and most of the melanin is finally excreted from the body as the stratum corneum falls off. Therefore, according to the formation, transportation and metabolism (excretion) process of skin melanin, the method of eliminating and reducing pigmentation may include: inhibiting the generation of melanin, such as controlling exogenous stimuli such as reducing ultraviolet radiation, or adding tyrosine structural analogs, inhibiting the binding of tyrosinase to tyrosine, inhibiting the activity of tyrosinase, etc.; or inhibiting the transport of melanin to keratinocytes; or accelerating the transfer of melanin to the stratum corneum, and strengthening the metabolism of the stratum corneum, thereby accelerating the metabolism of melanin. Among these possible methods, there are a variety of possible raw materials and their combinations, however, due to the mutual influence or restriction of various factors (such as the large side effects of hydroquinone, the poor solubility of azelaic acid, the difficulty in preparing a suitable preparation, the poor stability of vitamin C, and the high irritation of niacinamide), high-performance superior products are still scarce in the market.

[0004] At present, there are a variety of anti-aging methods and products, among which external products such as cosmetics are more convenient than products that need to be injected or taken orally. There are also many products on the market that focus on reducing pigmentation and fine lines. However, these products are limited by the performance of the raw materials themselves or the differences in formulas, and have more or less defects, such as insufficient safety and stability of the products, insufficient efficacy of the products to achieve the desired purpose, difficulty in obtaining raw materials for the products, poor operability and controllability of the products in production, etc. Therefore, it is still necessary and difficult to study external products that can achieve the desired elimination, reduction of skin pigmentation, reduction of fine lines or prevention of fine line formation in terms of efficacy, safety and stability. Summary of the invention

[0005] Based on the current problems and needs, the present invention provides a kit that can eliminate, alleviate and / or improve skin aging phenomena, such as dryness, roughness, pigmentation, fine lines, etc.; and also provides a kit with better stability, lower irritation and higher compliance.

[0006] Therefore, the present invention provides a kit, which comprises: a solid and a liquid; the solid is composed of a cocrystal of hydroxytyrosol and nicotinamide and trehalose, and the molar ratio of hydroxytyrosol to nicotinamide in the cocrystal is 1:1; the liquid comprises: oligopeptide-1, oligopeptide-2, oligopeptide-5, arginine / lysine polypeptide, acetyl octapeptide-3, dipeptide diaminobutyryl benzylamide diacetate, glycerol glucoside, oat β-glucan, p-hydroxyacetophenone, 1,3-butanediol, 1,2-hexanediol and water; the solid and the liquid are in a physically separated state before use.

[0007] The kit provided by the present invention adopts a physical separation state of solid and liquid before use, which is conducive to maintaining each material. Before use, the solid and the liquid are mixed and dissolved, and then applied to the facial skin or other skin surfaces in need.

[0008] Hydroxytyrosol, chemically known as 3,4-dihydroxyphenylethanol, has a strong antioxidant effect, but due to the presence of multiple hydroxyl groups in its structure, it has a low melting point and strong hygroscopicity, which is not conducive to the preparation of preparations. Niacinamide can inhibit the transport of melanin to keratinocytes, reduce the transfer of melanin to surface cells, and promote the shedding of keratinocytes containing melanin. It is often used in cosmetics, but it is more toxic and irritating at higher levels, and its whitening effect is difficult to meet the requirements at lower levels; in addition, niacinamide raw materials often contain a certain amount of nicotinic acid, and niacinamide is easily converted into hydrochloric acid under light, high temperature, acidic conditions, etc., and niacin will cause undesirable effects such as redness, swelling and stinging when used on the skin. Although hydroxytyrosol and niacinamide are commonly used chemical raw materials, they have disadvantages such as poor stability and high irritation in their own performance. When hydroxytyrosol, niacinamide and other auxiliary materials are mixed to form liquid or semi-solid external preparations, the products are easily unstable and discolor, stratify and / or increase impurities. In this case, in order to solve the problems of poor stability, high irritation and difficulty in effective absorption of the products, the method commonly used in the existing methods is to add a large amount of stabilizing ingredients and auxiliary ingredients that reduce irritation. However, the use of these large amounts of auxiliary ingredients easily leads to disadvantages such as complicated formulas, reduced safety, relatively reduced content of main functional ingredients and weakened product efficacy.

[0009] After research, the inventors found that the co-crystal form of hydroxytyrosol and niacinamide can effectively solve the problem of poor stability of the raw materials, and can improve their solubility and bioavailability, and more efficiently exert their functional effects; at the same time, adding trehalose can make the co-crystal more stable, and make the absorption of hydroxytyrosol and niacinamide more controllable after the use of the co-crystal, and the low irritation can effectively reduce.

[0010] Adding an appropriate amount of trehalose to the solid can further improve the stability of the eutectic and reduce the irritation of hydroxytyrosol and nicotinamide. The addition of an appropriate amount of trehalose not only improves the stability of the eutectic, but also makes the kit have a moisturizing and soothing effect, reduces the damage of epidermal cells due to external stimuli, significantly reduces irritation, and significantly improves tolerance. The level of trehalose has an important influence on the stability, irritation and use compliance of the kit. Although its content is too high, it helps to improve stability and reduce irritation, but it will greatly reduce the use compliance, and there is a strong skin stickiness and foreign body feeling when using it. If the content is too low, it cannot play a good role in improving stability and reducing irritation. In order to enhance the stability, efficacy, reduce irritation and improve use compliance of the kit, the mass ratio of the eutectic to trehalose needs to be controlled within an appropriate range. According to an embodiment of the present invention, the mass ratio of the eutectic to trehalose can be 1:0.5-1:5. In some embodiments, the mass ratio of the eutectic to trehalose is 1:1-1:5, which is conducive to reducing irritation. In some embodiments, the mass ratio of the co-crystal to trehalose is 1:2-1:4, which is beneficial to enhance the efficacy of the kit, reduce irritation, and improve compliance. In some embodiments, the mass ratio of the co-crystal to trehalose is 1:3, which is beneficial to enhance the efficacy of the kit, reduce irritation, and improve compliance.

[0011] While using trehalose to improve stability and efficacy and reduce and control the irritation of the kit, in order to further reduce and control irritation, the content of nicotinic acid in the co-crystal can be controlled. The higher the purity of the co-crystal, the more conducive it is to the efficacy of the kit and to reduce irritation and side effects. The inventors found that in the co-crystal or solid, the content of nicotinic acid is controlled to be less than 80ppm, which is more conducive to reducing irritation and facilitating production and quality control; when the nicotinic acid content is higher than 80ppm, the efficacy and stability of the co-crystal or solid are not only affected, but also the irritation and side reaction incidence rate of the kit after use increases, and the intensity of irritation and side effects also increases. In some embodiments, the content of nicotinic acid in the co-crystal or solid is less than 50ppm, which is more conducive to reducing irritation and improving compliance with use.

[0012] The efficacy of the kit is not only related to the components and their composition, but also to the content ratio of each component. In the present invention, in order to make the kit provide both efficient anti-aging effects and undesirable effects such as high stability and low irritation, it is necessary to control each component within a certain content range. According to an embodiment of the present invention, the ratio of the mass (mg) of the solid to the volume (mL) of the liquid can be 10mg / mL-200mg / ml. In some embodiments, the ratio of the mass (mg) of the solid to the volume (mL) of the liquid is 20mg / mL-200mg / ml, 40mg / mL-200mg / ml, 40mg / mL-100mg / ml, or 60mg / mL, or 80mg / mL, or 120mg / ml. In some embodiments, the ratio of the mass (mg) of the solid to the volume (mL) of the liquid is 20mg / mL-120mg / mL, which is conducive to providing both efficient anti-aging effects and maintaining high stability and low irritation. In some embodiments, the ratio of the mass (mg) of the solid to the volume (mL) of the liquid is 80 mg / mL, which is beneficial for providing efficient anti-aging effects while maintaining high stability and low irritation as well as a good user experience.

[0013] The liquid includes a variety of peptides and solvents, etc., which can dissolve the solids and allow hydroxytyrosol and niacinamide to be better absorbed; the combination of these peptides and solvents is beneficial to providing nutrients to the skin, helping to moisturize and absorb moisture for the skin, stimulate and promote the formation of elastin, and can improve skin elasticity and reduce fine lines. Moreover, these liquids and the solids can play a synergistic role after being combined. Compared with a single solid aqueous solution or the liquid used alone, the combined reagent has lower irritation and stronger effects on improving pigmentation, improving fine lines and other aging conditions, and is more conducive to anti-aging.

[0014] According to an embodiment of the present invention, in the liquid, based on the total mass of the liquid, the mass fraction of the oligopeptide-1, oligopeptide-2, oligopeptide-5 or arginine / lysine polypeptide does not exceed 0.002%, the mass fraction of the acetyl octapeptide-3 does not exceed 0.04%, or the mass fraction of the dipeptide diaminobutyryl benzylamide diacetate does not exceed 0.1%; if the content of these components is too high, the safety risk and quality control difficulty are greatly increased, and it is not suitable for the kit of the present invention; if it is too low, it is difficult to play the corresponding role.

[0015] In some embodiments, based on the total mass of the liquid, the mass fraction of oligopeptide-1 in the liquid is 0.0001%, the mass fraction of oligopeptide-2 is 0.00002%, the mass fraction of oligopeptide-5 is 0.00004%, the mass fraction of arginine / lysine polypeptide is 0.0002%, the mass fraction of acetyl octapeptide-3 is 0.003%, and the mass fraction of dipeptide diaminobutyryl benzylamide diacetate is 0.008%.

[0016] Based on the total mass of the liquid, in the liquid, the mass fraction of the glycerol glucoside may be 0.5%-5%, and the mass fraction of the oat β-glucan may be 0.5%-5%.

[0017] Based on the total mass of the liquid, in the liquid, the mass fraction of p-hydroxyacetophenone may be 0.1%-1%, the mass fraction of 1,3-butanediol may be 1%-10%, and the mass fraction of 1,2-hexanediol may be 0.1%-2%.

[0018] In some embodiments, based on the total mass of the liquid, in the liquid, the mass fraction of the glycerol glucoside is 1%, the mass fraction of the oat β-glucan is 1%, the mass fraction of the 1,3-butanediol is 5%, the mass fraction of the 1,2-hexanediol is 0.6%, and the mass fraction of the p-hydroxyacetophenone is 0.5%.

[0019] According to an embodiment of the present invention, the liquid of the kit optionally contains soluble collagen cross-linked polymer, sodium hyaluronate or zinc hyaluronate, 1,2-pentanediol, glycerol, glycerophosphoinositol choline salt, phenoxyethanol or ethylhexylglycerol; the liquid may contain one or more of these ingredients, or may not contain one or more of these ingredients.

[0020] In some embodiments, the liquid may contain soluble collagen cross-linked polymers in an amount of 0.01%-0.2% by mass based on the total mass of the liquid. In some embodiments, the liquid may contain soluble collagen cross-linked polymers in an amount of 0.15% by mass based on the total mass of the liquid.

[0021] Sodium hyaluronate or zinc hyaluronate can make the kit have a moisturizing effect, which is beneficial to moisturizing and improving dryness and roughness, and can be used as a thickener. In some embodiments, based on the total mass of the liquid, the liquid can contain sodium hyaluronate or zinc hyaluronate, and the mass fraction can be 0.01%-0.1%. In some embodiments, the liquid contains sodium hyaluronate, and the mass fraction is 0.05%. The sodium hyaluronate or zinc hyaluronate molecular weight can be 1 million Daltons-1.5 million Daltons.

[0022] In some embodiments, the liquid may contain 1,2-pentanediol in an amount of 0.01%-0.5% by mass based on the total mass of the liquid. In some embodiments, the liquid may contain 1,2-pentanediol in an amount of 0.1% by mass based on the total mass of the liquid.

[0023] In some embodiments, the liquid may contain glycerol in an amount of 0.2%-5% by mass based on the total mass of the liquid. In some embodiments, the liquid may contain glycerol in an amount of 2.2% by mass based on the total mass of the liquid.

[0024] In some embodiments, the liquid may contain glycerophosphoinositol choline salt in a mass fraction of 0.01%-0.5% based on the total mass of the liquid. In some embodiments, the liquid contains glycerophosphoinositol choline salt in a mass fraction of 0.2% based on the total mass of the liquid.

[0025] In some embodiments, the liquid may contain phenoxyethanol or ethylhexylglycerin at a mass fraction of 0.0001%-0.05% based on the total mass of the liquid. In some embodiments, the liquid contains phenoxyethanol or ethylhexylglycerin at a mass fraction of 0.002% based on the total mass of the liquid.

[0026] In some embodiments, based on the total mass of the liquid, the mass fraction of oligopeptide-1 in the liquid is 0.0001%, the mass fraction of oligopeptide-2 is 0.00002%, the mass fraction of oligopeptide-5 is 0.00004%, the mass fraction of arginine / lysine polypeptide is 0.0002%, the mass fraction of acetyl octapeptide-3 is 0.003%, and the mass fraction of dipeptide diaminobutyryl benzylamide diacetate is 0.008%; the mass fraction of the glycerol glucoside is 1%, and the mass fraction of the oat β-glucan is 1%; the mass fraction of p-hydroxyacetophenone is 0.5%, the mass fraction of 1,3-butanediol is 5%, and the mass fraction of 1,2-hexanediol is 0.6%.

[0027] In some embodiments, based on the total mass of the liquid, the mass fraction of oligopeptide-1 in the liquid is 0.0001%, the mass fraction of oligopeptide-2 is 0.00002%, the mass fraction of oligopeptide-5 is 0.00004%, the mass fraction of arginine / lysine polypeptide is 0.0002%, the mass fraction of acetyl octapeptide-3 is 0.003%, and the mass fraction of dipeptide diaminobutyryl benzylamide diacetate is 0.008%; the mass fraction of the glycerol glucoside is 1%, and the mass fraction of the swallow The mass fraction of wheat β-glucan is 1%, the mass fraction of p-hydroxyacetophenone is 0.5%, the mass fraction of 1,3-butanediol is 5%, and the mass fraction of 1,2-hexanediol is 0.6%; it optionally contains 0.15% of soluble collagen cross-linked polymer, 0.05% of sodium hyaluronate, 0.1% of 1,2-pentanediol, 2.2% of glycerol, and / or 0.2% of glycerophosphoinositide choline.

[0028] In some embodiments, based on the total mass of the liquid, the mass fraction of oligopeptide-1 in the liquid is 0.0001%, the mass fraction of oligopeptide-2 is 0.00002%, the mass fraction of oligopeptide-5 is 0.00004%, the mass fraction of arginine / lysine polypeptide is 0.0002%, the mass fraction of acetyl octapeptide-3 is 0.003%, and the mass fraction of dipeptide diaminobutyryl benzylamide diacetate is 0.008%; the mass fraction of the glycerol glucoside is 1%, and the mass fraction of the oat β-glucan is 1%, the mass fraction of p-hydroxyacetophenone is 0.5%, the mass fraction of 1,3-butanediol is 5%, and the mass fraction of 1,2-hexanediol is 0.6%; optionally containing 0.15% by mass of soluble collagen cross-linked polymer, 0.05% by mass of sodium hyaluronate, 0.1% by mass of 1,2-pentanediol, 2.2% by mass of glycerol, 0.2% by mass of glycerophosphoinositol choline salt, and / or 0.002% by mass of phenoxyethanol or ethylhexylglycerol.

[0029] The kit provided by the present invention is in a physically separated state before use. When used, the solid and the liquid are mixed and dissolved, and then the obtained mixed solution can be applied to the skin surface in need. After the solid and the liquid are mixed, the obtained mixed solution needs to be used completely within no more than 48 hours, or the obtained mixed solution needs to be stored in a light-proof environment of 0°C-15°C. In order to better ensure the efficacy and ensure the safety of use, after the solid and the liquid are mixed, the obtained mixed solution can be used up in 2 times within 24 hours or less, and the remaining liquid after the first use can be stored in a light-proof environment of 0°C-15°C. In order to better ensure the efficacy and ensure the safety of use, in some embodiments, the kit provided by the present invention is a type that is discarded after one or two uses after the solid and the liquid are mixed. In some embodiments, the kit provided by the present invention is a type that is discarded 24 hours after the solid and the liquid are mixed. In some embodiments, the kit provided by the present invention is a type that is discarded 48 hours or 48 hours after the solid and the liquid are mixed.

[0030] The kit of the present invention has a good effect of eliminating, reducing and / or improving skin pigmentation and fine lines, and has a synergistic effect on promoting the formation of collagen and elastin in the skin, reducing fine lines, and anti-aging effects. It also has good stability, low irritation and high compliance in use, can effectively improve the aging phenomenon of the skin, and is easy to control quality during production, store and transfer.

[0031] On the other hand, the present invention also provides a method for preparing the kit.

[0032] A method for preparing the aforementioned kit, comprising: obtaining the solid; and

[0033] Under heating and stirring conditions, part of water, 1,3-butylene glycol and 1,2-hexanediol are mixed, and p-hydroxyacetophenone is added, and sodium hyaluronate or zinc hyaluronate is optionally added; then the remaining water, 1,3-butylene glycol and 1,2-hexanediol, oligopeptide-1, oligopeptide-2, oligopeptide-5, acetyl octapeptide-3, arginine / lysine polypeptide, dipeptide diaminobutyryl benzylamide diacetate, glycerol glucoside and oat β-glucan are added under temperature control and stirring conditions, and 1,2-pentanediol, glycerol, soluble collagen cross-linked polymer, glycerophosphoinositol choline salt, phenoxyethanol and / or ethylhexylglycerol are optionally added; the mixture is mixed evenly to obtain the liquid; optionally, the obtained liquid and solid are respectively filled into containers to obtain a kit.

[0034] The heating condition is controlled at a temperature of 35°C-95°C. In some embodiments, the heating condition is controlled at a temperature of 40°C-85°C, or 60°C-85°C; or 40°C-65°C, or 45°C-65°C. In some embodiments, the heating condition is controlled at a temperature of 85°C, or 65°C, or 45°C or 35°C.

[0035] The temperature control is to control the temperature of the mixed system to 15°C-50°C. In some embodiments, the temperature control is to control the temperature of the mixed system to 15°C-45°C, or 15°C-35°C, or 20°C-40°C, or 20°C-45°C, or 30°C-50°C.

[0036] In some embodiments, a method for preparing the aforementioned kit comprises: mixing part of water, 1,3-butanediol and 1,2-hexanediol under stirring conditions at 40°C-85°C, and then adding p-hydroxyacetophenone, and optionally adding sodium hyaluronate or zinc hyaluronate; then controlling the temperature at 15°C-50°C and adding the remaining water, 1,3-butanediol and 1,2-hexanediol under stirring conditions, and then adding oligopeptide-1, oligopeptide-2, oligopeptide-5, acetyl octapeptide-3, arginine / lysine polypeptide, dipeptide diaminobutyryl benzylamide diacetate, glycerol glucoside and oat β-glucan, and optionally adding 1,2-pentanediol, glycerol, soluble collagen cross-linked polymer, glycerophosphoinositol choline salt, phenoxyethanol and / or ethylhexylglycerol; mixing evenly to obtain the liquid.

[0037] In some embodiments, a method for preparing the aforementioned kit comprises: mixing part of water, 1,3-butanediol and 1,2-hexanediol at 60°C-85°C and stirring, and then adding p-hydroxyacetophenone, and optionally adding sodium hyaluronate or zinc hyaluronate; then controlling the temperature at 20°C-45°C and stirring, adding the remaining water, 1,3-butanediol and 1,2-hexanediol, and then adding oligopeptide-1, oligopeptide-2, oligopeptide-5, acetyl octapeptide-3, arginine / lysine polypeptide, dipeptide diaminobutyryl benzylamide diacetate, glycerol glucoside and oat β-glucan, and optionally adding 1,2-pentanediol, glycerol, soluble collagen cross-linked polymer, glycerophosphoinositol choline salt, phenoxyethanol and / or ethylhexylglycerol; mixing evenly to obtain the liquid.

[0038] The method for preparing the kit is simple, controllable, and easy to operate, and the kit can be easily obtained, and the components therein will not be degraded, denatured, or otherwise fail to meet quality requirements. DETAILED DESCRIPTION

[0039] The term “comprise” or “include” is an open expression, that is, including the contents specified in the present invention but not excluding other contents.

[0040] In the present invention, “consisting of…” means that inevitable impurities and / or water may be contained.

[0041] In the present invention, ppm means parts per million and is calculated based on mass.

[0042] In the present invention, "optionally" means that it may be present or absent.

[0043] In the description of this specification, the description with reference to the terms "one embodiment", "some embodiments", "some implementation methods", "specific examples", or "some examples" etc. means that the specific features, structures, materials or characteristics described in conjunction with the embodiment or example are included in at least one embodiment or example of the present invention. In this specification, the schematic representations of the above terms do not necessarily refer to the same embodiment or example. Moreover, the specific features, structures, materials or characteristics described may be combined in any one or more embodiments or examples in a suitable manner. In addition, those skilled in the art may combine and combine the different embodiments or examples described in this specification and the features of the different embodiments or examples, unless they are contradictory.

[0044] In the present invention, room temperature is ambient temperature, which is 20°C-30°C, or 22°C-28°C, or 25°C.

[0045] In order to enable those skilled in the art to better understand the technical solution of the present invention, some non-limiting embodiments are further disclosed below to further describe the present invention in detail.

[0046] In the following examples, the cocrystal is a cocrystal of hydroxytyrosol and nicotinamide, which can be purchased from the market or prepared by the method disclosed in Chinese patent CN113004198, the molar ratio of hydroxytyrosol to nicotinamide is 1:1, and the nicotinic acid is less than 50 ppm.

[0047] Example I-1: Screening of kit formulations

[0048] 1) Different liquid formulas:

[0049]

[0050] Liquid preparation method: under heating conditions of 60°C-70°C, stir and mix water (about 60% of the total amount), 1,3-butanediol (about 60% of the total amount), 1,2-hexanediol (about 60% of the total amount), and p-hydroxyacetophenone, and add sodium hyaluronate (if any); then control the temperature at 40°C-50°C, add the remaining water, 1,3-butanediol and 1,2-hexanediol; then add oligopeptide-1, oligopeptide-2, oligopeptide-5, arginine / lysine polypeptide, acetyl octapeptide-3, dipeptide diaminobutyryl benzylamide diacetate, glycerol glucoside, oat (AVENA SATIVA) β-glucan, and other components: soluble collagen cross-linked polymer, glycerophosphoinositol choline salt, 1,2-pentanediol or glycerol; mix evenly to obtain the liquid.

[0051] Mixing and usage of liquid and co-crystal: Formula 1: The solubility of the co-crystal and the product of Formula 1 is poor, and it is absorbed slowly when used topically on the back of the hand, and the skin feels sticky; Formula 2: The solubility of the co-crystal and the product of Formula 2 is good, and it is absorbed quickly when used topically on the back of the hand, but the texture is thin and not moisturizing enough; Formula 3: The solubility of the co-crystal and the product of Formula 3 is good, and it is light, smooth and moisturizing when used topically on the back of the hand; Based on the mixing with the co-crystal and the feeling of use (including but not limited to tingling, stickiness, penetration speed, etc.), it is believed that liquid Formula 3 is relatively more suitable for co-crystal.

[0052] 2) Formulas of different ratios of cocrystal and trehalose:

[0053]

[0054] According to formula 4-formula 10, the eutectic and trehalose were weighed respectively, and then mixed to obtain a solid. The obtained solid was dissolved in 1 mL of a liquid based on formula 3 to obtain a solid-liquid mixture, the properties of the obtained mixture were observed, and the obtained mixture was locally tried on the back of the hand; the results: the formula without trehalose had a relatively obvious tingling sensation; the formula with too much trehalose had a relatively large sticky sensation. Taking into account the stability of the solid (a mixture of eutectic and trehalose), the skin feeling after the solid and liquid were dissolved (including but not limited to tingling, stickiness, and penetration speed), etc., it is considered that formula 8 is more suitable.

[0055] 3) Formulations with different solid-liquid ratios

[0056]

[0057] The solid prepared according to Formula 8 and the liquid prepared according to Formula 3 were mixed and dissolved according to the formula ratio to obtain each formula. Formula 11 dissolved faster, while Formulas 14 and 15 dissolved slower and were more viscous; considering the dissolution, viscosity, skin feeling during use (including but not limited to tingling, stickiness, penetration speed) and efficacy, Formulas 11-15 were considered acceptable, and Formulas 11-13 were more suitable than Formulas 14-15.

[0058] Example I-2: Preparation of kit

[0059] Formula: Solid: eutectic 1.8g, trehalose 5.4g; Liquid: 90mL;

[0060] Composition of the liquid: Based on the total mass of the liquid, the mass fraction of oligopeptide-1 is 0.0001%, the mass fraction of oligopeptide-2 is 0.00002%, the mass fraction of oligopeptide-5 is 0.00004%, the mass fraction of arginine / lysine polypeptide is 0.0002%, the mass fraction of acetyl octapeptide-3 is 0.003%, the mass fraction of dipeptide diaminobutyryl benzylamide diacetate is 0.008%; the mass fraction of p-hydroxyacetophenone is 0.5%, the mass fraction of 1,3-butanediol is The mass fraction of the raw materials is 5%, the mass fraction of 1,2-hexanediol is 0.6%, the mass fraction of glyceryl glucoside is 1%, the mass fraction of oat β-glucan is 1%, the mass fraction of sodium hyaluronate is 0.05%, the mass fraction of soluble collagen cross-linked polymer is 0.15%, the mass fraction of 1,2-pentanediol is 0.1%, the mass fraction of glycerol is 2.2%, the mass fraction of glycerophosphoinositol choline salt is 0.2%, the mass fraction of phenoxyethanol is 0.01%, the mass fraction of ethylhexylglycerol is 0.01%, and the balance is water.

[0061] Preparation process:

[0062] 1) Preparing a solid: weighing the cocrystal and trehalose according to the dosage of the formula, mixing them to obtain a solid;

[0063] 2) preparing a liquid: according to the dosage of the formula, under heating conditions of 60°C-70°C, stirring and mixing: water (about 60% of the total amount), 1,3-butylene glycol (about 60% of the total amount), 1,2-hexanediol (about 60% of the total amount), sodium hyaluronate and parahydroxyacetophenone; then controlling the temperature at 40°C-45°C, adding the remaining water, 1,3-butylene glycol and 1,2-hexanediol; adding oligopeptide-1, oligopeptide-2, oligopeptide-5, arginine / lysine polypeptide, acetyl octapeptide-3, dipeptide diaminobutyryl benzylamide diacetate, glycerol glucoside, oat (AVENA SATIVA) β-glucan, soluble collagen cross-linked polymer, glycerophosphoinositol choline salt, phenoxyethanol, ethylhexylglycerol, 1,2-pentanediol and glycerol; mixing evenly to obtain the liquid;

[0064] 3) Filling the prepared solid and liquid separately to obtain a test kit. The obtained test kit has a good user experience.

[0065] Example I-3: Preparation of kit

[0066] Formula: Solid: eutectic 1.8g, trehalose 5.4g; Liquid: 90mL;

[0067] Composition of the liquid: based on the total mass of the liquid, the mass fraction of oligopeptide-1 is 0.0001%, the mass fraction of oligopeptide-2 is 0.00002%, the mass fraction of oligopeptide-5 is 0.00004%, the mass fraction of arginine / lysine polypeptide is 0.0002%, the mass fraction of acetyl octapeptide-3 is 0.003%, the mass fraction of dipeptide diaminobutyryl benzylamide diacetate is 0.008%; the mass fraction of p-hydroxyacetophenone is 0.5%, the mass fraction of 1,3-butanediol is 5%, the mass fraction of 1,2-hexanediol is 0.6%, the mass fraction of glycerol glucoside is 1%, the mass fraction of oat β-glucan is 1%, and the balance is water.

[0068] Preparation process:

[0069] 1) Preparing a solid: weighing the cocrystal and trehalose according to the dosage of the formula, mixing them to obtain a solid;

[0070] 2) preparing a liquid: according to the dosage of the formula, under heating conditions of 60°C-70°C, stirring and mixing: water (about 60% of the total amount), 1,3-butanediol (about 60% of the total amount), 1,2-hexanediol (about 60% of the total amount) and p-hydroxyacetophenone; then controlling the temperature at 40°C-45°C, adding the remaining water, 1,3-butanediol and 1,2-hexanediol; adding oligopeptide-1, oligopeptide-2, oligopeptide-5, arginine / lysine polypeptide, acetyl octapeptide-3, dipeptide diaminobutyryl benzylamide diacetate, glycerol glucoside and oat (AVENASATIVA) β-glucan; mixing evenly to obtain the liquid;

[0071] 3) Filling the prepared solid and liquid separately to obtain a test kit. The obtained test kit has a good user experience.

[0072] Example II-1: Test on the inhibition rate of tyrosine kinase activity of cocrystal

[0073] The zebrafish test method was used to first test the maximum detection concentration (MTC) of the cocrystal. On this basis, zebrafish were used to test the inhibitory tyrosine kinase activity of the cocrystal to confirm whether the cocrystal can inhibit tyrosine kinase activity and the inhibition rate. Wild-type AB strain zebrafish were used, and the fish age was 6 hours after fertilization (6 hpf). Adult fish were raised and bred according to laboratory standard feeding and breeding methods, in compliance with the requirements of the international AAALAC certification.

[0074] 1) Randomly select zebrafish in a 6-well plate, 15 per well; dissolve the eutectic in water according to the set concentration (mass fraction), and set up a normal control group (water, no eutectic), with a volume of 3 mL per well; incubate at 28 °C in the dark for 45 h; determine the MTC of the eutectic on normal zebrafish based on the death and toxicity of zebrafish. The results of the MTC experiment are shown in the table below.

[0075]

[0076] According to the experimental results, the MTC of the cocrystal to normal zebrafish was measured to be 0.25%.

[0077] 2) Randomly select zebrafish in a 6-well plate, 30 per well; dissolve the test sample in water according to the set concentration (mass fraction), and set up a normal control group (without test sample) at the same time, with a volume of 3 mL per well, and three biological replicates; incubate at 28 °C in the dark for 45 h; grind the sample with a grinder, centrifuge and take the supernatant, and use the BCA protein concentration assay kit to determine the protein concentration of each experimental group; take 250 μg of total protein from zebrafish samples, add 1 mmol / L levodopa solution at a volume ratio of 1:1, and mix; transfer the liquid to a 96-well plate, 200 μL / well, and use an enzyme reader to measure the absorbance value (OD475) at a wavelength of 475 nm; calculate the inhibition rate of the sample's inhibition of tyrosine kinase activity according to the formula, inhibition rate (%) = (OD475 of the normal control group - OD475 of the sample group) / OD475 of the normal control group * 100%; statistical analysis p < 0.05 was considered to be significantly different; the results are shown in the table below.

[0078]

[0079] Note: SymWhite-377 refers to phenylethyl resorcinol.

[0080] According to the experimental results, the co-crystal can significantly inhibit the activity of tyrosine kinase.

[0081] In the following examples, the liquid (essence) is the liquid prepared according to Example I-2.

[0082] Example II-2: Irritation Test

[0083] 1) Chicken embryo chorioallantoic membrane eye irritation test:

[0084] Negative control group: 0.9% NaCl aqueous solution; Positive control group: 1% SDS (sodium dodecyl sulfate) aqueous solution; Take 0.3 mL respectively;

[0085] Sample group: Immediately before use, dissolve 100 mg of the eutectic in 3 ml of liquid (essence), and take 0.3 mL;

[0086] Ginseng root extract: Immediately before use, dissolve 100 mg of ginseng root extract in 3 ml of liquid (essence), and take 0.3 mL;

[0087] For SPF fertilized chicken embryos of White Leghorn chickens, on the 8th day of incubating the chicken embryos, use an egg candler to check the eggs, discard the unfertilized and inactive chicken embryos, select the chicken embryos with well-developed blood vessels, and mark the position of the air chamber on the eggshell surface; Start the experiment on the 9th day. Carefully peel off part of the eggshell of the air chamber with a dental curved forceps, drop a few drops of normal saline on the surface of the eggshell membrane to make it fully moist, pour out the liquid, and then carefully remove the eggshell membrane with forceps to ensure that the exposed chorioallantoic membrane is intact without any damage. Take 0.3 ml of the test substance and directly apply it to the chorioallantoic membrane (CAM), spread it as much as possible to ensure that the coverage area reaches at least 50%, after acting for 3 min, gently rinse the surface of the CAM with normal saline, complete the rinsing within 30 s, pour out the liquid, and immediately observe the degree of change of each toxic effect under a stereomicroscope and give a score (ES). The scoring criteria are shown in the following table.

[0088]

[0089] Scoring calculation formula: ES = sum of the degrees of bleeding, coagulation, and vascular lysis observed in 6 chicken embryos / 3; Conclusion judgment criteria: ES ≤ 4 is mild and non-irritating, 4 < ES ≤ 12 is mild irritation, 12 < ES < 16 is moderate irritation, ES ≥ 16 is severe irritation; The experimental results are shown in the following table.

[0090]

[0091] According to the experimental results, it can be known that the kit of the present invention is mild and non-irritating under this model.

[0092] 2) Subject use test:

[0093] Sample treatment: Immediately before use every morning, dissolve 100 mg of the eutectic in 3 ml of liquid (essence), then take an appropriate amount and apply it to the test area, and store the rest in a dark environment at 0 °C - 15 °C.

[0094] 30 healthy subjects (regardless of whether they have allergic skin or not) were selected, aged 20-40 years old, with an average age of 28 years old. The forehead area (same area) was selected, and the test sample was used once in the morning and evening every day after cleansing for 7 consecutive days. Every day, observe whether there are allergic phenomena such as redness, swelling, itching, tingling, rash or other abnormalities, and analyze and record them (if severe allergies or abnormalities occur, stop using it immediately, clean the face with water, and take treatment measures when necessary). The test results are as follows: 1 person had a slight tingling sensation when using it for the first time, which disappeared soon, and there was no abnormality in subsequent use; 1 person had a slight redness on the skin when using it for the first 3 days, which disappeared soon, and there was no abnormality in subsequent use; other users had no abnormalities during the whole process.

[0095] Example II-3: Elastase inhibition test

[0096] Sample: Just before use, dissolve 240 mg of solid (containing 60 mg of cocrystal and 180 mg of trehalose) in 3 ml of liquid (essence) and take 0.3 mL.

[0097] The principle of the elastase inhibition experiment is that elastase reacts with the enzyme substrate, and the absorbance changes after the active substance is added. The inhibition rate of the elastase inhibitor can be reflected by measuring the magnitude of the absorbance change.

[0098] The experiment was designed and conducted with reference to the "In vitro test method for anti-wrinkle and firming efficacy of cosmetics (elastase inhibition method)": appropriate amount of buffer solution, sample solution or control solution, and substrate were added to a 96-well plate, incubated for 20 minutes after oscillation mixing, and then elastase solution was added, and immediately incubated for 10 minutes after oscillation. The absorbance was measured at 410 nm using an ELISA reader, and the inhibition rate of elastase by the sample to be tested was calculated after analysis and processing of the data; Elastase inhibition rate (%) = [1-(AB) / (CD)] *100%, where:

[0099] A is the absorbance of the experimental group (containing substrate, buffer solution, elastase solution, and sample solution to be tested);

[0100] B is the absorbance of the experimental blank group (containing substrate and buffer solution, but not elastase solution, and containing the sample solution to be tested);

[0101] C is the absorbance of the control group (containing substrate, buffer solution, elastase solution, but not the sample solution to be tested);

[0102] D is the absorbance of the blank group (containing substrate and buffer solution, but not containing elastase solution and sample solution to be tested);

[0103] The experimental results are shown in the table below.

[0104]

[0105] Note: p<0.05 indicates that there is a statistical difference compared with the negative control group, and p≥0.05 indicates that there is no statistical difference compared with the negative control group.

[0106] According to the experimental results, the kit of the present invention can effectively inhibit elastase and has a firming and anti-wrinkle effect.

[0107] Example II-4: Stability test

[0108] The test was designed and carried out with reference to the Guiding Principles for Stability Testing of APIs and Preparations in the Appendix of Part IV of the 2020 edition of the Chinese Pharmacopoeia. The cocrystals and the solids obtained from Formula 8 (a mixture of cocrystals and trehalose, with a mass ratio of 1:3) were taken to investigate the stability of each test sample. The specific conditions are as follows: High temperature test (high temperature 60°C): The sample was placed in a brown light-proof glass bottle, sealed, and placed at 60±2°C, 75±5%RH. Samples were taken on 0 days and 7 days respectively, and the samples were observed and tested.

[0109] Test results: The content of hydroxytyrosol in the eutectic sample decreased slightly after 7 days, while the content of hydroxytyrosol in the sample obtained by formula 8 remained unchanged, indicating that the addition of trehalose can effectively protect the stability of hydroxytyrosol in the eutectic.

[0110]

[0111] The scheme of the present invention has been described through preferred embodiments, and relevant personnel can obviously modify or appropriately change and combine the scheme or application described herein within the content and scope of the present invention to implement and apply the technology of the present invention. Those skilled in the art can refer to the content of this article and appropriately improve the conditions / parameters to implement and / or apply the technology of the present invention. It is particularly important to point out that all similar replacements and modifications are obvious to those skilled in the art, and they are all considered to be included in the present invention.

Claims

1. A kit comprising: solid and liquid; of which, The solid is composed of a co-crystal of hydroxytyrosol and nicotinamide and trehalose, the mass ratio of the co-crystal to the trehalose is 1:3, and the molar ratio of hydroxytyrosol to nicotinamide in the co-crystal is 1:1; The liquid comprises: oligopeptide-1, oligopeptide-2, oligopeptide-5, arginine / lysine polypeptide, acetyl octapeptide-3, dipeptide diaminobutyryl benzylamide diacetate, glyceryl glucoside, oat β-glucan, p-hydroxyacetophenone, 1,3-butylene glycol, 1,2-hexanediol and water; the liquid optionally contains soluble collagen cross-linked polymer, sodium hyaluronate or zinc hyaluronate, 1,2-pentanediol, glycerol, glycerophosphoinositol choline salt, phenoxyethanol and / or ethylhexylglycerin; and The solid and the liquid are in a physically separated state before use, and the ratio of the mass of the solid to the volume of the liquid is 10 mg / mL-200 mg / ml.

2. The kit according to claim 1, wherein the content of niacin in the co-crystal or solid is less than 50 ppm.

3. The kit according to claim 2, wherein the ratio of the mass of the solid to the volume of the liquid is 80 mg / ml.

4. The kit according to claim 3, wherein in the liquid, based on the total mass of the liquid, the mass fraction of the oligopeptide-1, oligopeptide-2, oligopeptide-5 or arginine / lysine polypeptide does not exceed 0.002%, the mass fraction of the acetyl octapeptide-3 does not exceed 0.04%, or the mass fraction of the dipeptide diaminobutyryl benzylamide diacetate does not exceed 0.1%; the mass fraction of the glycerol glucoside is 0.5%-5%, the mass fraction of the oat β-glucan is 0.5%-5%, the mass fraction of the p-hydroxyacetophenone is 0.1%-1%, the mass fraction of the 1,3-butanediol is 1%-10%, and the mass fraction of the 1,2-hexanediol is 0.1%-2%.

5. The kit according to claim 4, based on the total mass of the liquid, in the liquid, the mass fraction of oligopeptide-1 is 0.0001%, the mass fraction of oligopeptide-2 is 0.00002%, the mass fraction of oligopeptide-5 is 0.00004%, the mass fraction of arginine / lysine polypeptide is 0.0002%, the mass fraction of acetyl octapeptide-3 is 0.003%, and the mass fraction of dipeptide diaminobutyryl benzylamide diacetate is 0.008%; the mass fraction of the glycerol glucoside is 1%, the mass fraction of the oat β-glucan is 1%, the mass fraction of p-hydroxyacetophenone is 0.5%, the mass fraction of 1,3-butanediol is 5%, and the mass fraction of 1,2-hexanediol is 0.6%.

6. The kit according to claim 5, optionally containing: 0.15% by mass of soluble collagen cross-linked polymer, 0.05% by mass of sodium hyaluronate, 0.1% by mass of 1,2-pentanediol, 2.2% by mass of glycerol, 0.2% by mass of glycerophosphoinositol choline, and / or 0.002% by mass of phenoxyethanol or ethylhexylglycerol.

7. The kit according to claim 6, wherein the kit is in a form that is discarded after being used once or twice after the solid and the liquid are mixed, or the kit is in a form that is discarded after 24 hours or 48 hours after the solid and the liquid are mixed.

8. A method for preparing the kit according to any one of claims 1 to 7, comprising: Obtaining the solid; and Under heating and stirring conditions, part of water, 1,3-butylene glycol and 1,2-hexanediol are mixed, and p-hydroxyacetophenone is added, and sodium hyaluronate or zinc hyaluronate is optionally added; then the remaining water, 1,3-butylene glycol and 1,2-hexanediol, oligopeptide-1, oligopeptide-2, oligopeptide-5, acetyl octapeptide-3, arginine / lysine polypeptide, dipeptide diaminobutyryl benzylamide diacetate, glycerol glucoside and oat β-glucan are added under temperature control and stirring conditions, and 1,2-pentanediol, glycerol, soluble collagen cross-linked polymer, glycerophosphoinositol choline salt, phenoxyethanol and / or ethylhexylglycerol are optionally added; and the mixture is mixed evenly to obtain the liquid; The obtained liquid and solid are respectively filled into containers to obtain a test kit; Wherein, the heating condition is to control the temperature to 35°C-95°C; and the temperature control is to control the temperature of the mixed system to 15°C-50°C.

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