Whitening composition, whitening product, preparation method and application

Abstract

The invention provides a whitening composition, a whitening product, a preparation method and application. The invention provides application of the milk exosome and nicotinamide in improving the autophagy ability of tranexamic acid activated melanocytes. The whitening composition provided by the invention is prepared from the following components in parts by weight: 1 to 50 parts of milk exosome, 5 to 40 parts of tranexamic acid and 0.5 to 30 parts of nicotinamide. The provided whitening composition is an active composition capable of activating the autophagy function of melanocytes of a human body, melanin synthesis in the melanocytes can be reduced by inducing generation of autophagy-related protein and activating an autophagy system, dullness of the skin is improved, the skin color is brightened through an internal channel, and the product is free of irritation and high in safety.

Description

Technical Field

[0001] The present invention relates to the technical field of cosmetics, and in particular to a whitening composition, a whitening product, a preparation method and an application thereof. Background Art

[0002] Melanin is a protein present in the basal layer of everyone's skin. Melanin is formed by a series of complex physiological processes in organisms and is mainly used to protect human skin cells. Therefore, when the skin reacts to external light, inflammation, oxidation, pollution, etc., the pre-existing melanin in the melanocytes is immediately redistributed, and then the melanin production in the epidermal melanocytes increases, and the melanin is transferred to the epidermal keratinocytes, forming the skin problems we see, such as spots and uneven skin color. Skin whitening requires regulating the melanin in the skin.

[0003] Currently, whitening products mainly reduce melanin content by inhibiting melanin synthesis, transport and metabolism. However, chemicals or drugs that can play a whitening role often need to be in high concentrations and large doses to be effective, which is very irritating to the skin. This strong stimulation may make the skin sensitive, causing the stratum corneum to become thinner, and then cause other new skin problems. How to achieve excellent whitening effects while reducing the irritation of whitening products is a technical problem that needs to be solved in the field of whitening product development. Summary of the invention

[0004] In view of the problems in the prior art, a whitening composition, a whitening product, a preparation method and an application are provided.

[0005] In some embodiments, provided is the use of milk exosomes and nicotinamide in improving the ability of tranexamic acid to activate melanocyte autophagy.

[0006] In some embodiments, a whitening composition is provided, comprising the following components, measured in parts by weight: 1 to 50 parts of milk exosomes, 5 to 40 parts of tranexamic acid, and 0.5 to 30 parts of niacinamide.

[0007] In some embodiments, the whitening composition comprises the following components in parts by weight: 3 to 25 parts of milk exosomes, 10 to 35 parts of tranexamic acid, and 0.8 to 15 parts of niacinamide.

[0008] In some embodiments, the method for preparing milk exosomes comprises the following steps: collecting whey in milk, mixing the collected whey with water, adjusting the pH to acidic to precipitate protein, separating the solid and liquid, collecting the supernatant, centrifuging the collected supernatant to collect the precipitate, and obtaining milk exosomes.

[0009] In some embodiments, the method for preparing milk exosomes satisfies one or more of the following characteristics:

[0010] (1) In the step of collecting whey from milk, the milk is centrifuged at 2° C. to 10° C. and a centrifugal force of 2000 g to 3000 g for 20 min to 30 min to separate the fat layer and the whey, and the whey is collected;

[0011] (2) In the step of mixing the collected whey with water, the volume ratio of the whey to the water is 1:(1-3);

[0012] (3) In the step of adjusting the pH to acidic to precipitate protein, the pH is adjusted to 4.58-4.74;

[0013] (4) In the solid-liquid separation step, the solid-liquid separation is performed by centrifugation at 2°C to 10°C and a centrifugal force of 10,000 g to 12,000 g for 40 min to 60 min;

[0014] (5) In the step of collecting the precipitate by centrifugation, the supernatant is centrifuged at 2°C to 10°C and a centrifugal force of 100,000 g to 150,000 g for 60 min to 90 min to collect the precipitate.

[0015] In some embodiments, the whitening composition further comprises one or more of a polyol and water.

[0016] In some embodiments, the whitening composition satisfies one or more of the following characteristics:

[0017] (1) The whitening composition contains 30 to 60 parts of polyol;

[0018] (2) The whitening composition contains 10 to 24 parts of water;

[0019] (3) The polyol includes one or more of pentanediol, ethylhexylglycerin, glycerol, hexanediol, and butylene glycol.

[0020] In some embodiments, a method for preparing a whitening composition is provided, comprising the following steps:

[0021] Mixing milk exosomes, tranexamic acid and niacinamide to prepare the whitening composition;

[0022] Wherein, when the whitening composition further includes one or more of polyols and water, the step of mixing milk exosomes, tranexamic acid and niacinamide further includes the step of mixing one or more of polyols and water.

[0023] In some embodiments, provided is the use of the whitening composition or the whitening composition prepared by the preparation method in preparing a whitening product.

[0024] In some embodiments, a whitening product is provided, wherein the whitening product comprises the whitening composition or the whitening composition prepared by the preparation method.

[0025] In some embodiments, the mass percentage of the whitening composition in the whitening product is 1% to 20%.

[0026] In some embodiments, the whitening product further comprises one or more of a moisturizer, a thickener, a skin conditioner, a preservative and water.

[0027] In some embodiments, the whitening product includes one or more of lotion, essence, gel, emulsion, cream, facial cleanser and facial mask.

[0028] The whitening composition provided in the present application is an active composition that can activate the autophagy function of human melanocytes, can reduce melanin synthesis in melanocytes by inducing the production of autophagy-related proteins and activating the autophagy system, and improves dull skin and brightens skin tone through internal pathways. The product is non-irritating and highly safe. Using it in the development of cosmetics has broad market application prospects. BRIEF DESCRIPTION OF THE DRAWINGS

[0029] In order to more clearly illustrate the technical solutions in the implementation methods and examples of the present application and to more completely understand the present application and its beneficial effects, the following is a brief introduction to the drawings required for use in the description of the implementation methods or examples. Obviously, the drawings described below are only some embodiments of the present application, and those skilled in the art can obtain other drawings based on these drawings without creative work.

[0030] Figure 1 The effects of the whitening compositions of Examples 1-2 and Comparative Examples 1-2 on the relative expression rate of LC3 protein;

[0031] Figure 2 The effects of the whitening compositions of Examples 1-2 and Comparative Examples 1-2 on the relative expression rate of Beclin-1 protein;

[0032] Figure 3The figures are VISIA diagrams of the skin status of volunteers using the whitening compositions of Examples 1 to 2 and Comparative Examples 1 to 2, wherein A1 is a VISIA diagram of the skin status of volunteers using the whitening composition of Example 1 on day 0, A2 is a VISIA diagram of the skin status of volunteers using the whitening composition of Example 1 on day 28, B1 is a VISIA diagram of the skin status of volunteers using the whitening composition of Example 2 on day 0, B2 is a VISIA diagram of the skin status of volunteers using the whitening composition of Example 2 on day 28, C1 is a VISIA diagram of the skin status of volunteers using the whitening composition of Comparative Example 1 on day 0, C2 is a VISIA diagram of the skin status of volunteers using the whitening composition of Comparative Example 1 on day 28, D1 is a VISIA diagram of the skin status of volunteers using the whitening composition of Comparative Example 2 on day 0, and D2 is a VISIA diagram of the skin status of volunteers using the whitening composition of Comparative Example 2 on day 28. DETAILED DESCRIPTION

[0033] In order to facilitate the understanding of the present invention, the present invention will be described more fully below with reference to the relevant drawings. The preferred embodiments of the present invention are given in the drawings. However, the present invention can be implemented in many different forms and is not limited to the embodiments described herein. On the contrary, the purpose of providing these embodiments is to make the understanding of the disclosure of the present invention more thorough and comprehensive.

[0034] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as those commonly understood by those skilled in the art to which the present invention belongs. The terms used herein in the specification of the present invention are only for the purpose of describing specific embodiments and are not intended to limit the present invention. The term "and / or" used herein includes any and all combinations of one or more of the related listed items.

[0035] Unless otherwise specified or incompatible herewith, the terms and phrases used herein shall have the following meanings:

[0036] The terms "and / or", "or / and", and "and / or" used in this application include any one of two or more related listed items, and also include any and all combinations of related listed items, and the arbitrary and all combinations include any two related listed items, any more related listed items, or a combination of all related listed items. It should be noted that when at least three items are connected by at least two conjunctions selected from "and / or", "or / and", and "and / or", it should be understood that in this application, the technical solution undoubtedly includes technical solutions that are all connected by "logical and", and undoubtedly includes technical solutions that are all connected by "logical or". For example, "A and / or B" includes three parallel solutions of A, B and "a combination of A and B".

[0037] In the present application, "plurality", "multiple", "multiple times", "multiples", etc., unless otherwise specified, refer to a number greater than 2 or equal to 2. For example, "one or more" means one or greater than or equal to two.

[0038] The terms "combination thereof", "any combination thereof", "any combination thereof" and the like used in this application include all suitable combinations of any two or more of the listed items.

[0039] In this application, the "suitable" mentioned in "suitable combination", "suitable method", "any suitable method", etc. is based on the ability to implement the technical solution of this application, solve the technical problems of this application, and achieve the expected technical effects of this application.

[0040] In the present application, "preferred", "better", "more preferred" and "suitable" are only used to describe implementation methods or examples with better effects. It should be understood that they do not constitute limitations on the scope of protection of the present application.

[0041] In the present application, "further", "further", "particularly" and the like are used for descriptive purposes to indicate differences in content, but should not be construed as limiting the scope of protection of the present application.

[0042] In this application, "optionally", "optional", and "optional" mean optional or dispensable, that is, any one of the two parallel schemes of "yes" or "no". If multiple "options" appear in a technical solution, unless otherwise specified and there is no contradiction or mutual restriction, each "optional" is independent.

[0043] In the present invention, in the "first aspect", "second aspect", "third aspect", "fourth aspect", etc., the terms "first", "second", "third", "fourth", etc. are only used for descriptive purposes and cannot be understood as indicating or implying relative importance or quantity, nor can they be understood as implicitly indicating the importance or quantity of the indicated technical features. Moreover, "first", "second", "third", "fourth", etc. only serve the purpose of non-exhaustive enumeration and description, and it should be understood that they do not constitute a closed limitation on quantity.

[0044] In the present application, the technical features described in an open manner include closed technical solutions composed of the listed features, and also include open technical solutions containing the listed features.

[0045] In the present application, when it comes to numerical intervals (i.e., numerical ranges), unless otherwise specified, the optional numerical distribution is considered continuous within the above numerical interval, and includes the two numerical endpoints (i.e., the minimum and maximum values) of the numerical range, and each numerical value between the two numerical endpoints. Unless otherwise specified, when the numerical interval only refers to the integers within the numerical interval, it includes the two endpoint integers of the numerical range, and each integer between the two endpoints. In this article, it is equivalent to directly listing each integer, such as t is an integer selected from 1 to 10, indicating that t is any integer selected from the group of integers consisting of 1, 2, 3, 4, 5, 6, 7, 8, 9 and 10. In addition, when multiple ranges are provided to describe features or characteristics, these ranges can be combined. In other words, unless otherwise specified, the ranges disclosed herein should be understood to include any and all subranges included therein.

[0046] The temperature parameters in this application, unless otherwise specified, are allowed to be either constant temperature treatment or to vary within a certain temperature range. It should be understood that the constant temperature treatment allows the temperature to fluctuate within the accuracy range controlled by the instrument. Fluctuations within the range of ±5°C, ±4°C, ±3°C, ±2°C, and ±1°C are allowed.

[0047] In the present application, % (w / w) and wt% both represent weight percentage, % (v / v) refers to volume percentage, and % (w / v) refers to mass volume percentage.

[0048] The "room temperature" in this application generally refers to 5°C to 30°C, preferably 25±5°C.

[0049] Skin whitening is a care project from the inside out. Currently, most whitening products focus on the synthesis, transport and metabolism of melanin to whiten the skin. However, the characteristics of chemicals or drugs that can achieve whitening effects are high concentration, high dosage and strong stimulation. Therefore, these strong stimulations may cause the skin to become sensitive, the skin stratum corneum is weak, and other new skin problems may occur. Therefore, by improving the body's own skin metabolism level to enhance the metabolism of melanin, the stimulation of excessive external whitening substances on the skin can be reduced.

[0050] During the autophagy process in the human body, some damaged proteins or organelles are wrapped by autophagic vesicles with a double membrane structure and sent to lysosomes for degradation and recycling. The cellular autophagy system plays an important role in removing waste proteins (misfolded or aggregated proteins) and eliminating dysfunctional organelles (including mitochondria, endoplasmic reticulum and peroxisomes). Under stress conditions, autophagy can be activated to form autophagosomes that engulf and degrade melanosomes. By binding to lysosomes, they can engulf black bodies and reduce melanin levels. When the external stimulus disappears, the synthesis of melanin will be reduced or terminated through regulation. At this time, melanin or melanosomes are cellular garbage that can be eliminated or should be eliminated under certain conditions. The main function of autophagy is to remove cellular garbage, and inducing autophagy can reduce melanin levels. In general, the goal of cells is to maintain homeostasis. Homeostasis is a kind of balance. For melanin homeostasis, it is to adjust melanin to the specific concentration required by cells in a specific environment by synthesizing or degrading melanin, and autophagy is a tool to achieve homeostasis. Then solving pigmentation problems through autophagy is a safe and effective method.

[0051] In some embodiments, provided is the use of milk exosomes and nicotinamide in improving the ability of tranexamic acid to activate melanocyte autophagy.

[0052] Tranexamic acid can activate the autophagy of melanocytes. The inventors of the present application have found that nicotinamide and milk exosomes can improve the efficiency of tranexamic acid in activating the autophagy of melanocytes. Milk exosomes contain a wealth of information and functional substances, such as more than 300 known miRNAs. Milk exosomes are rich in extracellular vesicles secreted by mammary epithelial cells, have a phospholipid bilayer membrane structure similar to that of cell membranes, and contain active substances such as proteins and nucleic acids. Studies have shown that miR-2478 encapsulated in milk exosomes can participate in signal transduction in vivo through a variety of cellular pathways. In the present invention, if milk exosomes are replaced with ordinary milk extracts, their function of improving the autophagy efficiency of tranexamic acid will be greatly reduced. At the same time, nicotinamide is added. Nicotinamide is a component of coenzyme I (NAD+) and coenzyme II (NADP+), and coenzymes are key substances in the process of cellular energy metabolism. These coenzymes play a key role in the redox reaction in cells, thereby participating in the energy metabolism process, helping to improve energy and generate efficiency.

[0053] In some embodiments, a whitening composition is provided, comprising the following components, measured in parts by weight: 1 to 50 parts of milk exosomes, 5 to 40 parts of tranexamic acid, and 0.5 to 30 parts of niacinamide.

[0054] In some embodiments, the whitening composition comprises the following components in parts by weight: 3 to 25 parts of milk exosomes, 10 to 35 parts of tranexamic acid, and 0.8 to 15 parts of niacinamide.

[0055] It can be understood that the whitening composition provided, in parts by weight, includes 1 to 50 parts of milk exosomes, which means that the weight parts of milk exosomes can be, for example, 1 part, 2 parts, 3 parts, 4 parts, 5 parts, 6 parts, 7 parts, 8 parts, 9 parts, 10 parts, 11 parts, 12 parts, 13 parts, 14 parts, 15 parts, 16 parts, 17 parts, 18 parts, 19 parts, 20 parts, 21 parts, 22 parts, 23 parts, 24 parts, 25 parts, 26 parts, 27 parts, 28 parts, 29 parts, 30 parts, 31 parts, 32 parts, 33 parts, 34 parts, 35 parts, 36 parts, 37 parts, 38 parts, 39 parts, 40 parts, 41 parts, 42 parts, 43 parts, 44 parts, 45 parts, 46 parts, 47 parts, 48 ​​parts, 49 parts, 50 parts, etc., or a range composed of any two of the foregoing values.

[0056] It can be understood that the whitening composition provided, measured in parts by weight, includes 5 to 40 parts of tranexamic acid, which means that the weight parts of tranexamic acid can be, for example, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, etc., or a range selected from any two of the foregoing values.

[0057] It can be understood that the whitening composition provided, in parts by weight, includes 0.5 parts to 30 parts of niacinamide, which means that the weight parts of niacinamide can be, for example, 0.5 parts, 1 parts, 2 parts, 3 parts, 4 parts, 5 parts, 6 parts, 7 parts, 8 parts, 9 parts, 10 parts, 11 parts, 12 parts, 13 parts, 14 parts, 15 parts, 16 parts, 17 parts, 18 parts, 19 parts, 20 parts, 21 parts, 22 parts, 23 parts, 24 parts, 25 parts, 26 parts, 27 parts, 28 parts, 29 parts, 30 parts, etc., or a range selected from any two of the foregoing values.

[0058] The whitening composition provided contains milk exosomes, tranexamic acid and niacinamide, which is an active composition that can activate the autophagy function of human melanocytes. It can reduce melanin synthesis in melanocytes by inducing the production of autophagy-related proteins and activating the autophagy system, and improve dull skin and brighten skin tone through internal pathways. The product is non-irritating and highly safe. It can be used in the development of cosmetics and has broad market application prospects.

[0059] In the whitening composition of the present invention, tranexamic acid can activate the autophagy ability of melanocytes, and nicotinamide and milk exosomes can improve the efficiency of tranexamic acid in activating the autophagy of melanocytes. Milk exosomes contain rich information and functional substances, such as more than 300 known miRNAs. Milk exosomes are rich in extracellular vesicles secreted by mammary epithelial cells, have a phospholipid bilayer membrane structure similar to that of cell membranes, and contain active substances such as proteins and nucleic acids. Studies have shown that miR-2478 encapsulated in milk exosomes can participate in signal transduction in vivo through a variety of cellular pathways. In the present invention, if milk exosomes are replaced with ordinary milk extracts, the function of improving the autophagy efficiency of tranexamic acid will be greatly reduced. At the same time, nicotinamide is added, which is a component of coenzyme I (NAD+) and coenzyme II (NADP+), and coenzymes are key substances in the process of cellular energy metabolism. These coenzymes play a key role in the redox reaction in cells, thereby participating in the energy metabolism process, helping to improve energy and generate efficiency.

[0060] The milk exosomes in the whitening composition provided are obtained by skimming and removing protein from fresh milk or pasteurized milk to obtain whey, which is concentrated through multiple centrifugation and ultrafiltration processes to obtain a milk extract rich in milk exosome particles.

[0061] In some embodiments, the method for preparing milk exosomes comprises the following steps: collecting whey in milk, mixing the collected whey with water, adjusting the pH to acidic to precipitate protein, separating the solid and liquid, collecting the supernatant, centrifuging the collected supernatant to collect the precipitate, and obtaining milk exosomes.

[0062] In some embodiments, the method for preparing milk exosomes comprises the following steps:

[0063] S100: collects whey from milk;

[0064] S200: mixing the collected whey with water, adjusting the pH to acidic to precipitate protein, separating the solid and the liquid, and collecting the supernatant;

[0065] S300: The collected supernatant is centrifuged to collect the precipitate to obtain milk exosomes.

[0066] In some embodiments, in the method for preparing milk exosomes, in step S100, the milk is centrifuged at 2°C to 10°C and a centrifugal force of 2000g to 3000g for 20 to 30 minutes to separate the fat layer and the whey, and the whey is collected.

[0067] In some embodiments, in the method for preparing milk exosomes, in step S200, the volume ratio of whey to water is 1:(1-3), for example, the volume ratio of whey to water can be 1:1, 1:2, 1:3 or a range selected from any two of the foregoing ratios.

[0068] In some embodiments, in the method for preparing milk exosomes, in step S200, the pH is adjusted to 4.58-4.74, for example, the pH is adjusted to 4.58, 4.59, 4.60, 4.61, 4.62, 4.63, 4.64, 4.65, 4.66, 4.67, 4.68, 4.69, 4.70, 4.71, 4.72, 4.73, 4.74 or a range selected from any two of the foregoing values.

[0069] In some embodiments, in the method for preparing milk exosomes, in step S200, solid-liquid separation is performed by centrifugation at a centrifugal force of 10000 g to 12000 g for 40 min to 60 min at 2° C. to 10° C.

[0070] In some embodiments, in the method for preparing milk exosomes, in step S300, the collected supernatant is centrifuged at 2° C. to 10° C. and a centrifugal force of 100,000 to 150,000 g for 60 min to 90 min to collect the precipitate.

[0071] In some embodiments, the method for preparing milk exosomes comprises the following steps:

[0072] S100: Whey collection: centrifuge the milk at 2°C~10°C and a centrifugal force of 2000g~3000g for 20 min~30 min to separate the fat layer from the whey and collect the whey;

[0073] S200: supernatant collection: add water to the collected whey, with the volume ratio of whey to water being 1:(1-3), adjust the pH to 4.58-4.74 with acid solution to precipitate protein, then centrifuge at 2°C-10°C and a centrifugal force of 10,000-12,000 g for 40-60 minutes, and collect the supernatant after centrifugation;

[0074] S300: Collection of milk exosomes: Centrifuge the system obtained in step S200 twice, take the supernatant, and centrifuge at 2°C~10°C and a centrifugal force of 100000g~150000g for 60min~90min to obtain the precipitate to obtain milk exosomes.

[0075] In some embodiments, the whitening composition further comprises one or more of a polyol and water.

[0076] In some embodiments, the whitening composition contains 30 to 60 parts of polyol.

[0077] In some embodiments, the whitening composition contains 10 parts to 24 parts of water.

[0078] In some embodiments, the polyol includes one or more of pentylene glycol, ethylhexylglycerin, glycerol, hexylene glycol, and butylene glycol.

[0079] In some embodiments, a method for preparing a whitening composition is provided, comprising the following steps: mixing milk exosomes, tranexamic acid and niacinamide to prepare a whitening composition;

[0080] Wherein, when the whitening composition further includes one or more of polyols and water, the step of mixing milk exosomes, tranexamic acid and niacinamide further includes the step of mixing one or more of polyols and water.

[0081] In some embodiments, a use of a whitening composition in preparing a whitening product is provided.

[0082] In some embodiments, a whitening product is provided, comprising a whitening composition.

[0083] In some embodiments, the mass percentage of the whitening composition in the whitening product is 1% to 20%. For example, the mass percentage of the whitening composition in the whitening product can be 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20% or a range selected from any two of the foregoing values.

[0084] In some embodiments, the whitening product further comprises one or more of a moisturizer, a thickener, a skin conditioner, a preservative and water.

[0085] In some embodiments, the moisturizer in the whitening product comprises one or more of polyol, sodium hyaluronate, polyamino acid polysaccharide condensate, β-glucan, sodium PCA, and betaine.

[0086] In some embodiments, the polyol includes one or more of pentylene glycol, ethylhexylglycerin, glycerol, hexylene glycol, and butylene glycol.

[0087] In some embodiments, the thickener in the whitening product comprises one or more of cellulose, starch, sodium polyacrylate, sodium alginate, carbomer, acrylates, and polysaccharides.

[0088] In some embodiments, the skin conditioning agent in the whitening product comprises one or more of allantoin, dipotassium glycyrrhizinate, vitamin C, lipoic acid, glabridin, ascorbic acid derivatives, α-arbutin, nonapeptide-1, astaxanthin, and phenethylresorcinol.

[0089] In some embodiments, the preservatives in the whitening product include one or more of parabens, chlorphenesin, phenoxyethanol, potassium sorbate, and sodium benzoate.

[0090] In some embodiments, the whitening product includes one or more of a lotion, an essence, a gel, an emulsion, a cream, a facial cleanser, and a facial mask.

[0091] The present invention is further explained below in conjunction with specific embodiments, but the present invention is not limited to these embodiments.

[0092] The tranexamic acid used in the examples was purchased from Hunan Dongting Pharmaceutical Co., Ltd.; niacinamide was purchased from Royal DSM of the Netherlands; milk was purchased from Inner Mongolia Mengniu Dairy (Group) Co., Ltd.; and acrylic acid (esters) / C10-30 alkyl acrylate cross-linked polymer was purchased from Lubrizol.

[0093] 1. Preparation of whitening composition

[0094] Embodiment 1 A whitening composition

[0095] 30 parts of 1,2-pentanediol, 1 part of ethylhexylglycerin, 15 parts of tranexamic acid, 1 part of niacinamide, 5 parts of milk exosomes, and 53 parts of purified water were mixed and stirred to completely dissolve them to obtain a whitening composition.

[0096] The preparation method of milk exosomes is as follows:

[0097] S100: Whey collection: Take 100 ml of fresh milk, centrifuge at 8°C, 3000g for 20 min, separate the fat layer from the whey, and collect the whey;

[0098] S200: supernatant collection: add an equal volume of 100 ml ultrapure water to the above whey, adjust the pH to 4.60 with acid solution while stirring to precipitate protein, and then centrifuge in a high-speed refrigerated centrifuge at 8°C and 11000g for 50 min, and collect the supernatant after centrifugation;

[0099] S300: Collection of milk exosomes: Centrifuge the system obtained in step S200 twice, take the supernatant, centrifuge at 130,000 g for 80 min at 8°C, and take the precipitate to obtain milk exosomes.

[0100] Embodiment 2 A whitening composition

[0101] 30 parts of 1,2-pentanediol, 1 part of ethylhexylglycerin, 30 parts of tranexamic acid, 10 parts of niacinamide, 20 parts of milk exosomes and 9 parts of purified water were mixed and stirred to completely dissolve them to obtain a whitening composition.

[0102] Among them, the preparation method of milk exosomes is the same as the preparation method of milk exosomes in Example 1.

[0103] Embodiment 3 A whitening composition

[0104] 30 parts of 1,2-pentanediol, 1 part of ethylhexylglycerin, 30 parts of tranexamic acid, 1 part of niacinamide, 5 parts of milk exosomes, and 33 parts of purified water were mixed and stirred to completely dissolve them to obtain a whitening composition.

[0105] Among them, the preparation method of milk exosomes is the same as the preparation method of milk exosomes in Example 1.

[0106] Comparative Example 1 A whitening composition

[0107] 30 parts of 1,2-pentanediol, 1 part of ethylhexylglycerin, 30 parts of tranexamic acid and 39 parts of purified water were mixed and stirred to completely dissolve them, thereby obtaining a control whitening composition.

[0108] Comparative Example 2 A whitening composition

[0109] 30 parts of 1,2-pentanediol, 1 part of ethylhexylglycerin, 50 parts of tranexamic acid and 19 parts of purified water were mixed and stirred to completely dissolve them, thereby obtaining a control whitening composition.

[0110] Comparative Example 3: a whitening composition

[0111] Similar to Example 2, except that the milk exosomes are replaced with an equal mass of milk.

[0112] Comparative Example 4: a whitening composition

[0113] 30 parts of 1,2-pentanediol, 1 part of ethylhexylglycerol, 30 parts of tranexamic acid, 30 parts of milk exosomes, and 9 parts of purified water were mixed and stirred to completely dissolve them to obtain a control composition.

[0114] Comparative Example 5: a whitening composition

[0115] 30 parts of 1,2-pentanediol, 1 part of ethylhexylglycerin, 30 parts of tranexamic acid, 30 parts of niacinamide, and 9 parts of purified water were mixed and stirred to completely dissolve them, thereby obtaining a control composition.

[0116] 2. Performance Test

[0117] The composition was prepared according to the following formula: 10% of the whitening composition of Examples 1 to 3 or Comparative Examples 1 to 5, 0.15% of acrylic acid (ester) / C10-30 alkyl acrylate cross-linked polymer, 0.5% of phenoxyethanol, 5% of glycerol, 0.10% of arginine, and 84.25% of purified water were taken by weight percentage. The raw materials were stirred evenly to obtain the test compositions of Examples 1 to 3 and Comparative Examples 1 to 5.

[0118] (1) Effect of whitening composition on autophagy

[0119] Western blotting was used to detect LC3 protein and Beclin-1 protein to observe whether the whitening composition had an effect on autophagy. SPSS 22.0 software was used for statistical analysis, and the results were expressed as X±SD, indicating statistical significance.

[0120] LC3 is a protein light chain 3 and a marker of the autophagic process. Its function is mainly involved in the formation of autophagosomes. The LC3 precursor molecule is cut off the C-terminal 5-peptide by ATG4B and cleaved to form the cytoplasmic form LC3-I. It is then activated by APG7L / ATG7, transferred to ATG3 and coupled with acylethanolamine (PE) to form the membrane-bound form LC3-II, which can attach to the membrane of the autophagosome and is a structural protein of the autophagosome.

[0121] Beclin-1 is a member of the autophagy-related protein family and is involved in regulating the formation of autophagic vesicles. It promotes the maturation of autophagic vesicles by interacting with multiple proteins, such as Vps34 and UVRAG.

[0122] Test method: Mouse melanoma cells (B16-F1) were cultured at 0.5×10 5Cells were seeded at a density of 100 μg / well in a 6-well culture plate and cultured in a cell incubator (37°C, 5% (v / v) CO2) for 24 hours. The cells were treated with 1 mg / ml of the test composition or 10 ng / ml of transforming growth factor-β (TGF-β) for 15 minutes to induce autophagy. The transforming growth factor-β treatment group was used as the positive control group. Protein samples were collected from the treated cells, and protein extracts were isolated from the cells by adding ProNA CETi lysis buffer, where the protein concentration was measured by Bradford assay. 50 μg of protein was added to 5 × loading buffer and mixed, boiled at 100°C for 10 min to completely denature the protein, and then the prepared samples were added to 8%, 12% or 15% polyacrylamide gels. After electrophoresis, the protein was transferred to a PVDF membrane, then blocked with 5% skim milk powder for 1 hour, and incubated with specific primary antibodies LC3 (dilution ratio) and Beclin-1 (dilution ratio) at 4°C overnight. The next day, wash with TBST (250 mM Tris-HCl, 1.5 mM NaCl and 0.1% Tween 20) buffer 3 times, 10 minutes each time. Dilute the corresponding HRP-conjugated secondary antibody (1:4000)) and incubate at room temperature for 2 hours, and wash the PVDF membrane with TBST buffer 3 times, 10 minutes each time. Mix the chemiluminescent reagent A solution and B solution in a 1:1 ratio and set aside. When using, draw an appropriate volume of luminescent droplets on the PVDF membrane, then develop it in a developing instrument and save the experimental pictures.

[0123] Figure 1 is the relative expression rate of LC3 protein, Figure 2 is the relative expression rate of Beclin-1 protein. Figure 1 It can be seen that compared with the level in the positive control group, the expression levels of LC3 in the mouse melanoma cells treated with the test compositions of Examples 1 to 3 and Comparative Examples 1 to 5 increased by approximately 48%, 190%, 89%, 42%, 156%, 60%, 93%, and 96%, respectively. Figure 2It can be seen that the expression levels of Beclin-1 protein in mouse melanoma cells treated with the test compositions of Examples 1 to 3 and Comparative Examples 1 to 5 increased by about 62%, 181%, 76%, 55%, 150%, 59%, 83%, and 87%, respectively, compared with the levels in the positive control group. These results show that both the Examples and Comparative Examples can induce the upregulation of autophagy-related proteins (Beclin-1 and LC3) in B16 F1 cells by activating the ERK1 / 2 signaling pathway. However, it can be seen from the comparison of Example 2 and Comparative Example 1 that the addition of milk exosomes and nicotinamide will increase the autophagy ability of tranexamic acid to activate melanocytes; compared with Example 1 and Example 3, a concentration-dependent relationship is presented. Nicotinamide and milk exosomes cooperate with each other to have a better effect than single nicotinamide or milk exosomes. Comparative Example 1 omits nicotinamide and milk exosomes relative to Example 2, and the autophagy ability of melanocytes is significantly deteriorated.

[0124] (2) Verification of skin efficacy and safety

[0125] 40 female volunteers with sensitive skin were selected, with an age distribution of 20 to 40 years old, and randomly divided into four groups A, B, C, and D, with 10 people in each group. Group A used the test composition of Example 1, Group B used the test composition of Example 2, Group C used the test composition of Comparative Example 1, and Group D used the test composition of Comparative Example 2. They were used once in the morning and evening each day, and compared after 28 days. The Derma Lab instrument was used to measure the skin's ΔL, ΔB, and ΔM values, and the data before 0 days were compared for analysis. VISIA was also used to compare the skin conditions. The results are shown in Tables 1 and Figure 3 .

[0126] Depend on Figure 3 From the VISIA comparison results, it can be seen that from the effect of volunteers' use, the data of the four groups of volunteers A, B, C, and D have all improved after 28 days of persistent use. Among them, the skin of more than 70% of the volunteers in group A has been significantly improved, and the skin glossiness has been improved. At the same time, the reduction rate of the area of ​​brown spots of volunteers is 40%, and none of the volunteers reported any discomfort symptoms. The skin of more than 80% of the volunteers in group B has been significantly improved, and the skin glossiness has been improved. At the same time, the reduction rate of the area of ​​brown spots of volunteers is 60%, and none of the volunteers reported any discomfort symptoms. The skin of more than 60% of the volunteers in group C has been significantly improved, and the skin glossiness has been improved. At the same time, the reduction rate of the area of ​​brown spots of volunteers is 10%. The skin of more than 80% of the volunteers in group D has been significantly improved, and the skin glossiness has been improved. At the same time, the reduction rate of the area of ​​brown spots of volunteers is 50%, but 1 volunteer reported discomfort.

[0127] Table 1 ΔL, ΔB and ΔM values ​​of volunteers

[0128]

[0129] From the data in Table 1, we can see that the content of tranexamic acid in group A is lower than that in group C, but the skin glossiness, ΔL value and ΔB value of volunteers in group A are not much different from those in group C; compared with the high content of tranexamic acid in group D, the safety of volunteers in group B is higher than that in group D under the premise of equal effect. Therefore, we can see that niacinamide and milk exosomes can further promote the autophagy of tranexamic acid for melanin, and can achieve excellent whitening effect under the premise of reducing the amount of tranexamic acid used and reducing the irritation of tranexamic acid to the skin.

[0130] The technical features of the above-described embodiments may be arbitrarily combined. To make the description concise, not all possible combinations of the technical features in the above-described embodiments are described. However, as long as there is no contradiction in the combination of these technical features, they should be considered to be within the scope of this specification.

[0131] The above-mentioned embodiments only express several implementation methods of the present invention, and the description thereof is relatively specific and detailed, but it cannot be understood as limiting the scope of the invention patent. It should be pointed out that for ordinary technicians in this field, several variations and improvements can be made without departing from the concept of the present invention, which all belong to the protection scope of the present invention. Therefore, the protection scope of the patent of the present invention shall be based on the attached claims, and the description and drawings may be used to interpret the content of the claims.

Claims

1. Application of milk exosomes and nicotinamide in improving the ability of tranexamic acid to activate melanocyte autophagy.

2. A whitening composition, characterized in that: The composition comprises the following components by weight: 1 to 50 parts of milk exosomes, 5 to 40 parts of tranexamic acid, and 0.5 to 30 parts of nicotinamide; Optionally, the following components are included: 3 to 25 parts of milk exosomes, 10 to 35 parts of tranexamic acid, and 0.8 to 15 parts of niacinamide.

3. The whitening composition according to claim 2, characterized in that The method for preparing milk exosomes comprises the following steps: collecting whey in milk, mixing the collected whey with water, adjusting the pH to acidic to precipitate protein, separating the solid from the liquid, collecting the supernatant, centrifuging the collected supernatant to collect the precipitate, and obtaining milk exosomes.

4. The whitening composition according to claim 3, characterized in that The method for preparing milk exosomes meets one or more of the following characteristics: (1) In the step of collecting whey from milk, the milk is centrifuged at 2°C to 10°C and a centrifugal force of 2000g to 3000g for 20 min to 30 min to separate the fat layer and the whey, and the whey is collected; (2) In the step of mixing the collected whey with water, the volume ratio of the whey to the water is 1:(1-3); (3) In the step of adjusting the pH to acidic to precipitate protein, the pH is adjusted to 4.58-4.74; (4) In the solid-liquid separation step, the solid-liquid separation is performed by centrifugation at 2°C to 10°C and a centrifugal force of 10,000 g to 12,000 g for 40 min to 60 min; (5) In the step of collecting the precipitate by centrifugation, the supernatant is centrifuged at 2°C to 10°C and a centrifugal force of 100,000 g to 150,000 g for 60 min to 90 min to collect the precipitate.

5. The whitening composition according to any one of claims 2 to 4, characterized in that The whitening composition further comprises one or more of a polyol and water; Optionally, the whitening composition satisfies one or more of the following characteristics: (1) The whitening composition contains 30 to 60 parts of polyol; (2) The whitening composition contains 10 to 24 parts of water; (3) The polyol includes one or more of pentanediol, ethylhexylglycerin, glycerol, hexanediol and butanediol.

6. A method for preparing a whitening composition, characterized in that: The following steps are involved: Mixing milk exosomes, tranexamic acid and niacinamide to prepare the whitening composition according to any one of claims 2 to 5; Wherein, when the whitening composition further includes one or more of polyols and water, the step of mixing milk exosomes, tranexamic acid and niacinamide further includes the step of mixing one or more of polyols and water.

7. Use of the whitening composition according to any one of claims 2 to 5 or the whitening composition prepared by the preparation method according to claim 6 in the preparation of whitening products.

8. A whitening product, characterized in that: The whitening product comprises the whitening composition according to any one of claims 2 to 5 or the whitening composition prepared by the preparation method according to claim 6; Optionally, the mass percentage of the whitening composition in the whitening product is 1% to 20%.

9. The whitening product according to claim 8, characterized in that: The whitening product also includes one or more of a moisturizer, a thickener, a skin conditioner, a preservative and water.

10. The whitening product according to claim 8 or 9, characterized in that: The whitening product comprises one or more of lotion, essence, gel, emulsion, cream, cleanser and mask.

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