Whitening composition suitable for sensitive skin staying up late and application thereof

Abstract

The invention provides a whitening composition suitable for sensitive skin staying up late. According to the present invention, the red, yellow and black skin color problem of people with sensitive skin staying up late is solved, and on the basis of inhibition of tyrosinase and inhibition of melanin synthesis, the anti-oxidation, anti-saccharification, inflammatory factor inhibition, skin rhythm regulation and other aspects are combined, such that skin erosion caused by the outside during the day is resisted, skin circadian rhythm is improved, and skin aging is inhibited. The composition effectively regulates and controls the damage of staying up late to the skin, improves the self-repairing ability of the skin at night, and forms an efficient whitening composition with daily prevention and night repairing synergistic whitening. Compared with a common whitening composition, the whitening composition is higher in whitening efficiency, and skin color problems such as red, yellow and black skin and the like are improved more comprehensively; the whitening composition provided by the invention is stable in property, is suitable for various types of cosmetic dosage forms, can be widely applied to various whitening cosmetics, and makes contributions to developing new fields related to skin whitening and expanding the market.

Description

Technical Field

[0001] The invention belongs to the technical field of cosmetics, and particularly relates to a whitening composition suitable for use on sensitive skin that stays up late, and an application thereof. Background Art

[0002] Staying up late has become a normal state due to the work and life patterns of modern people. Staying up late can cause a series of problems that affect the skin color, such as dull skin and yellow face. Currently, there are limited products on the market for staying up late skin, and there are insufficient products for long-term improvement of skin problems caused by staying up late, especially for dull skin, yellow skin, roughness and dullness caused by staying up late. Scientists have found that staying up late can cause skin circadian rhythm disorders, abnormal expression of clock factors such as BMAL1, BMAL1 knockout reduces the transcription of PER1, and PER1 silencing can induce phosphorylation of MITF, the master regulator of melanin production, thereby stimulating melanin production and increasing melanocyte activity. Experiments have shown that staying up late (sleeping only 4 hours a night for 5-6 consecutive days) will make the skin yellowness value higher and higher, and it will not recover immediately due to sleep recovery. At the same time, the rhythm disorder caused by staying up late can cause barrier damage and inflammatory response, making the skin sensitive. In the long run, it will eventually become sensitive skin, which is susceptible to external stimuli, leading to inflammatory pigmentation, making the skin dull.

[0003] The color of normal skin is determined by the content and distribution of various pigments, including hemoglobin (oxidized and reduced), carotene or carotenoids, collagen and melanin in the dermis, and is related to the thickness of the skin and the scattering of light on the skin surface. The main determining factor is the content and distribution of melanin.

[0004] Researchers have conducted a lot of research on whitening issues and have a relatively in-depth understanding of the mechanism of melanin production and deposition. Many results have been published on inhibiting melanin production. Currently, whitening products mostly inhibit melanin production or have relatively mature solutions for melanin transport and metabolism.

[0005] However, with the changes in the social environment and the fact that staying up late has become a common phenomenon, the daily erosion of the skin by the external environment and the disruption of skin rhythm caused by staying up late for a long time have caused many products to be effective in vitro but have poor clinical effects. Currently, there are limited products on the market for skin that stays up late, and there is a lack of products for long-term improvement of skin problems caused by staying up late, especially for the dull, yellow, rough and dull skin caused by staying up late. Most of the existing whitening products are designed from the perspective of melanin synthesis, transport and metabolism. There are few that simultaneously target sensitive skin caused by staying up late and solve the problem from the internal mechanism. There are comprehensive whitening products suitable for sensitive skin caused by staying up late, such as dark, red and dark yellow skin. There is a lack of overall comprehensive consideration, resulting in existing whitening products that only treat the symptoms but not the root cause.

[0006] Therefore, how to scientifically screen effective ingredients and how to reasonably build formulas to maximize whitening efficiency will become the key to solving the problem of whitening sensitive skin caused by staying up late. Summary of the invention

[0007] In view of this, the technical problem to be solved by the present invention is to provide a whitening composition suitable for sensitive skin caused by staying up late and its application. The present invention aims at the problems of dull, uneven and yellow skin tone in people with sensitive skin who stay up late, and provides a whitening composition from multiple aspects such as anti-oxidation + anti-glycation + anti-inflammatory (synergistic whitening, improving whitening efficiency), inhibiting tyrosinase, inhibiting melanin production, etc., which comprehensively solves the problems of red, yellow and black skin tone and achieves whitening effects from multiple dimensions.

[0008] The present invention provides a whitening composition suitable for use on sensitive skin caused by staying up late, comprising:

[0009] 0.005-5 parts by weight of skin biorhythm regulating factor;

[0010] Ascorbyl glucoside 0.02-3 parts by weight;

[0011] 0.01 to 2 parts by mass of potassium 4-methoxysalicylate;

[0012] 0.001 to 1 parts by weight of thyme extract;

[0013] Carnosine 0.002-1 parts by weight;

[0014] 0.001-1 parts by weight of Kava extract;

[0015] 0.01 to 2 parts by mass of citrus fruit extract.

[0016] Preferably, the whitening composition comprises:

[0017] Skin biorhythm regulating factor 0.005 to 1 parts by weight;

[0018] Ascorbyl glucoside 0.02-2 parts by mass;

[0019] 0.01 to 1 parts by mass of potassium 4-methoxysalicylate;

[0020] 0.001 to 1 parts by weight of thyme extract;

[0021] Carnosine 0.002-0.2 parts by mass;

[0022] 0.001-0.6 parts by weight of Kava extract;

[0023] 0.01 to 1 part by weight of citrus fruit extract.

[0024] Preferably, the whitening composition comprises:

[0025] 0.5 parts by weight of skin biorhythm regulator;

[0026] Ascorbyl glucoside 1.8 parts by mass;

[0027] 0.8 parts by mass of potassium 4-methoxysalicylate;

[0028] 1 part by weight of thyme extract;

[0029] Carnosine 0.15 mass parts;

[0030] 0.6 parts by weight of Kava extract;

[0031] 1 part by weight of citrus fruit extract;

[0032] Or, include:

[0033] Skin biorhythm regulating factor 0.015 parts by mass;

[0034] Ascorbyl glucoside 0.02 parts by mass;

[0035] 0.02 parts by mass of potassium 4-methoxysalicylate;

[0036] 0.01 parts by mass of thyme extract;

[0037] Carnosine 0.015 mass parts;

[0038] 0.01 parts by weight of Kava extract;

[0039] 0.01 parts by mass of citrus fruit extract;

[0040] Or, include:

[0041] Skin biorhythm regulating factor 0.005 parts by mass;

[0042] Ascorbyl glucoside 0.02 parts by mass;

[0043] 0.01 parts by mass of potassium 4-methoxysalicylate;

[0044] 0.01 parts by mass of thyme extract;

[0045] Carnosine 0.002 mass parts;

[0046] 0.006 parts by weight of Kava extract;

[0047] 0.01 parts by mass of citrus fruit extract.

[0048] Preferably, the whitening composition and the skin biorhythm regulating factor include a combination of yeast glycoprotein and amino acids, adenosine, and biorhythm peptides.

[0049] Preferably, the whitening composition and the skin biorhythm regulating factor include:

[0050] 0.002 to 2 parts by weight of a combination of yeast glycoprotein and amino acids;

[0051] 0.001 to 1 parts by weight of adenosine;

[0052] 0.002 to 2 parts by mass of biorhythm peptide.

[0053] Preferably, in the whitening composition, the kava extract is an extract of at least one of the leaves, roots and stems of Piper methysticum.

[0054] The present invention also provides an application of the whitening composition in whitening products.

[0055] The present invention also provides a cosmetic comprising the whitening composition.

[0056] Preferably, the whitening composition is added in an amount of 0.0049% to 20% in cosmetics.

[0057] Preferably, the cosmetic comprises:

[0058] 0.0049% to 20% whitening composition;

[0059] 2% to 10% moisturizer;

[0060] The balance is water.

[0061] Compared with the prior art, the present invention provides a whitening composition suitable for staying up late for sensitive skin, including: 0.005-5 mass parts of skin biorhythm regulator; 0.02-3 mass parts of ascorbyl glucoside; 0.01-2 mass parts of potassium methoxysalicylate; 0.001-1 mass parts of European thyme extract; 0.002-1 mass parts of carnosine; 0.001-1 mass parts of kava pepper extract; 0.01-2 mass parts of citrus fruit extract. The present invention solves the red, yellow and black skin color problems of people with sensitive skin who stay up late. On the basis of inhibiting tyrosinase and inhibiting melanin synthesis, it combines anti-oxidation, anti-glycation, inhibition of inflammatory factors, and regulation of skin rhythms to resist the erosion of the skin by the outside world during the day, improve the circadian rhythm of the skin, effectively regulate the damage to the skin caused by staying up late, and at the same time enhance the self-repair ability of the skin at night, forming a high-efficiency whitening composition of day-defense and night-repair synergistic whitening, and its synergistic efficiency and whitening effect are verified by biochemical experiments, cell tests, zebrafish tests, and human efficacy evaluation. Compared with general whitening compositions, this whitening composition has higher whitening efficiency and more comprehensive improvement on skin color problems such as red, yellow, and black skin. The whitening composition provided by the present invention has stable properties and is suitable for various types of cosmetic dosage forms. It can be widely used in various whitening cosmetics and contributes to the development of new fields related to skin whitening and the expansion of the market. BRIEF DESCRIPTION OF THE DRAWINGS

[0062] Figure 1 The following is a diagram showing the skin status after using the samples for different periods of time. DETAILED DESCRIPTION

[0063] The present invention provides a whitening composition suitable for use on sensitive skin caused by staying up late, comprising:

[0064] 0.005-5 parts by weight of skin biorhythm regulating factor;

[0065] Ascorbyl glucoside 0.02-3 parts by weight;

[0066] 0.01 to 2 parts by mass of potassium 4-methoxysalicylate;

[0067] 0.001 to 1 parts by weight of thyme extract;

[0068] Carnosine 0.002-1 parts by weight;

[0069] 0.001-1 parts by weight of Kava extract;

[0070] 0.01 to 2 parts by mass of citrus fruit extract.

[0071] The whitening composition provided by the present invention has the functions of protecting the skin from environmental damage during the day that causes the production of melanin, and regulating the skin's biological rhythm at night to promote the expression of rhythmic factors, thereby improving the skin's repair ability, improving dullness, and achieving a comprehensive whitening effect around the clock.

[0072] The whitening composition provided by the present invention, which is suitable for use on sensitive skin caused by staying up late, comprises: 0.005 to 5 parts by weight of a skin biorhythm regulating factor, which may be 0.005, 0.01, 0.02, 0.03, 0.04, 0.05, 0.7, 0.9, 1, 2, 3, 4, 5, or any value between 0.005 and 5 parts by weight.

[0073] The skin biorhythm regulating factor is a raw material composition capable of regulating skin rhythm and activating skin repair ability, including a composition of yeast glycoprotein and amino acids, adenosine, and biorhythm peptides. In some specific embodiments of the present invention, the skin biorhythm regulating factor is composed of three raw materials including a composition of yeast glycoprotein and amino acids, adenosine, and biorhythm peptides:

[0074] In some specific embodiments of the present invention, the skin biorhythm regulating factor comprises:

[0075] 0.002 to 2 parts by weight of a combination of yeast glycoprotein and amino acids;

[0076] 0.001 to 1 parts by weight of adenosine;

[0077] 0.002 to 2 parts by mass of biorhythm peptide.

[0078] Specifically, the skin biorhythm regulating factor includes 0.002 to 2 parts by mass of a composition of yeast glycoprotein and amino acids, and can be 0.002, 0.005, 0.01, 0.05, 0.1, 0.2, 0.3, 0.4, 0.5, 0.7, 0.9, 1, 1.1, 1.3, 1.5, 1.7, 1.9, 2, or any value between 0.01 and 2 parts by mass.

[0079] The INCI name of the composition of yeast glycoprotein and amino acids is: glycoprotein, glutamic acid, valine, threonine and water. The composition of yeast glycoprotein and amino acids comprises 3% glycoprotein, 3% glutamic acid, 0.55% valine, 0.55% threonine and the balance water.

[0080] The skin biorhythm regulating factor also includes 0.01 to 1 parts by mass of adenosine, which can be 0.01, 0.05, 0.1, 0.2, 0.3, 0.4, 0.5, 0.7, 0.9, 1, or any value between 0.01 and 1 parts by mass. Adenosine is an endogenous nucleoside that is found throughout human cells, and its full name is adenine nucleotide. Adenosine is a metabolite of adenosine triphosphate (ATP), which is the main substance that provides energy for human life activities. After dephosphorylation, it is converted into ADP, and then dephosphorylated to AMP, and then a phosphate is removed to become adenosine. In this process, a large amount of energy is released, and it is vividly called "life fuel".

[0081] By adding additional adenosine to the skin biorhythm regulator, cells are encouraged to synthesize more ATP, thereby generating more energy, promoting cell repair and metabolism. In cosmetics, adenosine can inhibit inflammatory responses by activating its receptors, and has the effects of promoting angiogenesis, promoting collagen synthesis, and accelerating wound healing.

[0082] The skin biorhythm regulating factor also includes 0.01 to 2 parts by mass of biorhythm peptides, which can be 0.01, 0.05, 0.1, 0.2, 0.3, 0.4, 0.5, 0.7, 0.9, 1, 1.1, 1.3, 1.5, 1.7, 1.9, 2, or any value between 0.01 and 2 parts by mass. The INCI name of the biorhythm peptide is glutamine ethyl imidazole. It can imitate the activation of circadian rhythm genes by sunlight, promote the expression of rhythmic genes, and adjust the reasonable state between the physiological activity of skin cells and environmental constraints. It helps repair skin cells from multiple dimensions such as improving cell defense, optimizing cell renewal, and improving skin microcirculation.

[0083] The skin biorhythm regulator is a composition of yeast glycoprotein and various amino acids extracted by fermenting brewer's yeast using biotechnology under aerobic conditions. It has the effect of providing enough energy to restore the skin to a normal state and helping cells to self-renew and self-repair. The mechanism of action is to promote mitochondrial oxygen consumption to increase the level of ATP, improve metabolism, and stimulate the expression of key genes to improve aerobic respiration, anaerobic glycolysis, and mitochondrial protection and regeneration.

[0084] The skin biorhythm regulator works from three dimensions: biological clock gene regulation + stimulation of the cell's own energy + additional energy supplementation. The various components cooperate with each other and promote each other, playing a good synergistic role. It is a combination with strong late-night skin care effects.

[0085] The whitening composition provided by the present invention, which is suitable for use on sensitive skin caused by staying up late, also includes 0.02 to 3 parts by mass of ascorbyl glucoside, which may be 0.02, 0.03, 0.04, 0.05, 0.1, 0.5, 1, 1.5, 1.8, 2, 2.5, 3, or any value between 0.02 and 3 parts by mass.

[0086] Ascorbyl glucoside (AA2G) is a less irritating ingredient among all VC and its derivatives. Ascorbyl glucoside can reduce melanin through reduction, and can effectively protect the skin from UV damage. It also has physiological activities such as anti-scurvy, antioxidant, and antiviral. It also has the effects of whitening, maintaining skin elasticity, and preventing skin inflammation and photoaging.

[0087] The whitening composition suitable for staying up late for sensitive skin provided by the present invention also includes 0.01 to 2 parts by mass of potassium 4-methoxysalicylate, which can be 0.01, 0.02, 0.03, 0.04, 0.05, 0.06, 0.07, 0.08, 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 1, 1.2, 1.4, 1.5, 1.6, 1.8, 2, or any value between 0.01 to 2 parts by mass. The whitening mechanism of potassium 4-methoxysalicylate (4MSK) covers multiple dimensions, including: inhibiting tyrosinase activity, inhibiting B16 cell melanin, anti-oxidation, inhibiting the decrease of loricrin caused by UVB, thereby reducing spots, etc. Studies have shown that 4MSK at a concentration of 1% can inhibit tyrosinase activity by 96.7%, 4MSK at a concentration of 0.1% can inhibit B16 cell melanin by 98.5%, 4MSK at a concentration of 0.6% can inhibit ABTS+ production by 93.2%, and 4MSK at a concentration of 1.0% can inhibit the reduction of loricrin caused by UVB, with an inhibitory activity of 7%.

[0088] The whitening composition suitable for staying up late for sensitive skin provided by the present invention also includes 0.001 to 1 parts by mass of European thyme extract, which can be 0.001, 0.005, 0.01, 0.5, 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1, or any value between 0.001 and 1 parts by mass. European thyme extract inhibits the activity of intracellular kinesin and prevents the transfer of melanin from pigment cells to epidermal keratinocytes, thereby reducing the content of melanin on the surface of the skin and achieving a whitening effect. The experimental results show that European thyme extract can significantly reduce the transfer of fluorescent particles in cells and inhibit melanin synthesis. 3D skin models and human clinical trials have confirmed its whitening effect.

[0089] The whitening composition suitable for staying up late for sensitive skin provided by the present invention also includes 0.002 to 1 parts by mass of carnosine, which can be 0.002, 0.005, 0.01, 0.015, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1, or any value between 0.002 and 1 parts by mass. Carnosine (L-Carnosine), scientific name β-alanyl-L-histidine, is a dipeptide composed of two amino acids, β-alanine and L-histidine, a crystalline solid, which participates in the formation of human muscle and eye and brain lens structure. It has the effects of anti-oxidation, anti-glycosylation, and anti-protein cross-linking at the molecular level, and the effects of delayed aging and immune activation at the cellular level.

[0090] The specific mechanism of action is as follows:

[0091] Carnosine has antioxidant properties and is a natural antioxidant. Carnosine has been proven to remove free radicals produced during oxidative stress, which are more damaging to the skin than sunlight. Free radicals are highly active atoms or groups of atoms in the human body that can oxidize other substances in the human body, causing excessive oxidation of fatty acids in cell membranes to form reactive oxygen free radicals (ROS) and α-β unsaturated aldehydes. Carnosine has a significant inhibitory effect on lipid oxidation caused by free radicals and metal ions. Its use in cosmetics can prevent skin aging and whiten the skin.

[0092] Carnosine has a good anti-glycation effect. Carbonylated proteins and advanced glycation end products (AGEs) are signals of cell aging. They accumulate abnormally in cells and accelerate cross-linking with other proteins. Carnosine can react with sugar before sugar erodes the skin, replacing the body's proteins in the body to react with sugar, thereby helping to protect proteins from being glycated and achieving an anti-glycation effect.

[0093] The whitening composition suitable for staying up late for sensitive skin provided by the present invention also includes 0.001 to 1 parts by mass of kava extract, which can be 0.001, 0.003, 0.005, 0.006, 0.008, 0.01, 0.02, 0.04, 0.05, 0.06, 0.08, 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1.0, or any value between 0.001 and 1 parts by mass. The kava extract is an extract of at least one of the leaves, roots, and stems of kava.

[0094] The main mechanism of action of the kava extract is to act on multiple pathway receptors of the nervous system (TRPV1, GABAA, CB1) through the synergistic effect of multiple kavalactones, thereby reducing nerve sensitivity. Specifically, kava extract has the following effects: inhibiting the production of TRPV1 receptor protein, relieving the discomfort after skin stimulation; activating the binding of GABA and GABAA receptors, enhancing the inhibition of nerve pain transmission, and playing a sedative role; affinity with CB1 receptors, playing an anti-anxiety effect. Through the above-mentioned multiple mechanisms of action, kava extract can reduce nerve sensitivity, thereby alleviating skin sensitivity symptoms and exerting a soothing and calming effect.

[0095] The whitening composition suitable for staying up late for sensitive skin provided by the present invention also includes 0.01 to 2 parts by mass of citrus fruit extract, which can be 0.01, 0.03, 0.05, 0.07, 0.09, 0.1, 0.3, 0.5, 0.7, 0.9, 1, 1.1, 1.3, 1.5, 1.7, 1.9, 2, or any value between 0.01 and 2 parts by mass. Citrus fruit extract is a skin "fire extinguisher" derived from natural plants and suitable for sensitive skin. The skin capsaicin receptor TRPV1 channel can be regulated to reduce the skin's discomfort to external stimuli by inhibiting the expression, activation and effect of TRPV1 receptors. It has been proven in human tests that it can effectively block the tingling discomfort caused by lactic acid and improve the skin's tolerance, thereby achieving the purpose of soothing the skin.

[0096] In some specific embodiments of the present invention, the whitening composition suitable for use on sensitive skin due to staying up late comprises:

[0097] Skin biorhythm regulating factor 0.005 to 1 parts by weight;

[0098] Ascorbyl glucoside 0.02-2 parts by mass;

[0099] 0.01 to 1 parts by mass of potassium 4-methoxysalicylate;

[0100] 0.001 to 1 parts by weight of thyme extract;

[0101] Carnosine 0.002-0.2 parts by mass;

[0102] 0.001-0.6 parts by weight of Kava extract;

[0103] 0.01 to 1 part by weight of citrus fruit extract.

[0104] In some preferred embodiments of the present invention, the whitening composition suitable for use on sensitive skin due to staying up late comprises:

[0105] 0.5 parts by weight of skin biorhythm regulator;

[0106] Ascorbyl glucoside 1.8 parts by mass;

[0107] 0.8 parts by mass of potassium 4-methoxysalicylate;

[0108] 1 part by weight of thyme extract;

[0109] Carnosine 0.15 mass parts;

[0110] 0.6 parts by weight of Kava extract;

[0111] 1 part by weight of citrus fruit extract;

[0112] In some preferred embodiments of the present invention, the whitening composition suitable for use on sensitive skin due to staying up late comprises:

[0113] Skin biorhythm regulating factor 0.015 parts by mass;

[0114] Ascorbyl glucoside 0.02 parts by mass;

[0115] 0.02 parts by mass of potassium 4-methoxysalicylate;

[0116] 0.01 parts by mass of thyme extract;

[0117] Carnosine 0.015 mass parts;

[0118] 0.01 parts by weight of Kava extract;

[0119] 0.01 parts by mass of citrus fruit extract;

[0120] In some preferred embodiments of the present invention, the whitening composition suitable for use on sensitive skin due to staying up late comprises:

[0121] Skin biorhythm regulating factor 0.005 parts by mass;

[0122] Ascorbyl glucoside 0.02 parts by mass;

[0123] 0.01 parts by mass of potassium 4-methoxysalicylate;

[0124] 0.01 parts by mass of thyme extract;

[0125] Carnosine 0.002 mass parts;

[0126] 0.006 parts by weight of Kava extract;

[0127] 0.01 parts by mass of citrus fruit extract.

[0128] The main determinant of skin color is the content and distribution of melanin. The melanin synthesis pathway is that melanocytes exist in the basal layer of the epidermis of the skin, with dendritic protrusions on the surface that extend all the way to the surrounding keratinocytes. Its cytoplasm contains melanosomes, which contain tyrosinase (TYR). Stimulated by ultraviolet rays, emotions, stress and other factors, TYR is activated and reacts with tyrosine in the blood with the help of oxygen to produce gray DOPA, which then generates black-gray dopaquinone (DQ), which is further oxidized, catalyzed and polymerized through different pathways to form melanin particles. Under the promotion of cell metabolism, it is transferred to the surface of the skin and cannot be metabolized in time, so it gradually accumulates and precipitates to form spots. The melanin metabolism pathway is that after the melanosomes are transferred from melanocytes to keratinocytes, as the keratinocytes differentiate and move upward to the epidermis, the protein structure of the melanosomes will be gradually degraded by lysosomes, and finally excreted from the body as the keratinocytes fall off.

[0129] Therefore, the main mechanism of whitening is to interfere with the production of pigments and the transfer process of pigment particles, which usually includes inhibiting tyrosinase activity, reducing tyrosinase synthesis, reducing tyrosinase migration, and producing specific toxicity to melanocytes.

[0130] The present invention aims to solve the problem of red, yellow and black skin color of people with sensitive skin who stay up late. On the basis of inhibiting tyrosinase and melanin synthesis, it combines anti-oxidation, anti-glycation, inhibition of inflammatory factors, and regulation of skin rhythm, to resist external erosion of the skin during the day, improve the skin's circadian rhythm, effectively regulate the damage of staying up late to the skin, and at the same time enhance the skin's self-repair ability at night, forming a synergistic whitening effect of day protection and night repair.

[0131] The composition provided by the present invention combines daytime defense and nighttime repair with the inhibition of melanin production, thereby improving the whitening effect in multiple dimensions from solving the problem of red, yellow and black skin.

[0132] The present invention also provides an application of the whitening composition in whitening products.

[0133] The present invention also provides a cosmetic comprising the whitening composition. The present invention has no particular restrictions on the product form of the cosmetic, which may be a toner, emulsion, cream, mask, essence, or gel.

[0134] In the present invention, the amount of the whitening composition added to the cosmetic is 0.0049% to 20%, which can be 0.0049%, 0.005%, 0.007%, 0.009%, 0.01%, 0.05%, 0.07%, 0.1%, 0.5%, 0.7%, 1%, 3%, 5%, 7%, 9%, 10%, 11%, 13%, 15%, 17%, 19%, 20%, or any value between 0.0049% and 20%.

[0135] In some specific embodiments of the present invention, the cosmetic comprises:

[0136] 0.0049% to 20% whitening composition;

[0137] 2% to 10% moisturizer;

[0138] The balance is water.

[0139] In the present invention, the cosmetic comprises 0.0049% to 20% of the whitening composition, which may be 0.0049%, 0.005%, 0.007%, 0.009%, 0.01%, 0.05%, 0.07%, 0.1%, 0.5%, 0.7%, 1%, 3%, 5%, 7%, 9%, 10%, 11%, 13%, 15%, 17%, 19%, 20%, or any value between 0.0049% and 20%.

[0140] The cosmetics also include 2% to 10% of a moisturizer, which may be 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, or any value between 2% and 10%. Preferably, the moisturizer is selected from one or more of 1,3-butylene glycol, glycerol, trehalose, betaine, xylitol, glyceryl glucoside, propylene glycol, glyceryl polyether-26, diglycerol, dipropylene glycol, and dipropylene glycol.

[0141] In addition to the above raw materials, the cosmetic also includes a balance of water.

[0142] In some specific embodiments of the present invention, the cosmetics may further include other types of additives, such as one or more of preservatives, thickeners, emulsifiers, and emollients, as needed. The types and amounts of the above additives must comply with the requirements of the Cosmetic Safety Technical Specifications.

[0143] In the present invention, the preservative is preferably one or more of p-hydroxyacetophenone and 1,2-hexanediol.

[0144] In order to further understand the present invention, the whitening composition suitable for sensitive skin caused by staying up late and its application provided by the present invention are described below in conjunction with embodiments. The protection scope of the present invention is not limited by the following embodiments.

[0145] Examples 1 to 3

[0146] 1. Formulation

[0147] Table 1 Recipe

[0148]

[0149] The composition of yeast glycoprotein and amino acids includes 3% glycoprotein, 3% glutamic acid, 0.55% valine, 0.55% threonine and the balance water.

[0150] 2. Preparation process

[0151] Dissolve the potassium 4-methoxysalicylate in the above raw materials with an appropriate amount of water at 80°C.

[0152] The remaining raw materials are mixed at 40-45° C., and the dissolved 4-methoxysalicylic acid potassium aqueous solution is added and mixed evenly.

[0153] Test Example 1

[0154] 1 Antioxidant Experiment

[0155] 1.1 Experimental Principle

[0156] Biological oxidation is the process in which hydrogen and electrons removed from metabolites are oxidized to synthesize water and release energy through a series of enzymatic reactions in the body. Redox reactions need to be carried out continuously in the life process to maintain the dynamic balance of oxidation. More than 95% of the oxygen that enters the body is converted into energy through the oxidation of food and reduced to water itself, and the remaining less than 5% of oxygen is reduced to oxygen free radicals (ROS). Under normal circumstances, the generation and removal of free radicals and reactive oxygen species are in a dynamic balance, and the presence of appropriate free radicals and reactive oxygen species is essential to the body. When the internal and external factors of the body break the dynamic balance of generation and removal, resulting in the large-scale generation and accumulation of free radicals and reactive oxygen species in the body, it will cause cell aging, apoptosis and even necrosis.

[0157] 1,1-Diphenyl-2-trinitrophenylhydrazine (DPPH for short) is a stable, long-lived free radical. Its ethanol solution is dark purple and has strong absorption near 517nm. When a free radical scavenger is present, the light absorption of the DPPH ethanol solution is weakened due to pairing with its single electron. The degree of fading of the DPPH ethanol solution is linearly related to the number of electrons it accepts, which can be used to evaluate the sample's ability to scavenge free radicals, that is, the strength of its antioxidant activity. Specifically, the strength of the antioxidant ability can be expressed by the scavenging rate. The larger the scavenging rate, the stronger the antioxidant activity.

[0158] 1.2 Experimental procedures

[0159] ① Prepare DPPH solution: Accurately weigh a certain amount of DPPH and use anhydrous ethanol to prepare a 0.1mM DPPH solution (the absorbance measured by an ELISA reader is about 0.7). The preparation should be done in a dark place and the prepared solution should be stored in a dark place.

[0160] ② Prepare experimental group solutions: Prepare sample solutions of a certain concentration according to Table 2.

[0161] Table 2 Experimental design

[0162] Experimental Group Solution 1 0.15% Carnosine aqueous solution 2 1.8% Ascorbyl Glucoside Aqueous Solution 3 0.8% Potassium 4-methoxysalicylate aqueous solution 4 0.15% Carnosine + 1.8% Ascorbyl Glucoside + 0.8% Potassium 4-Methoxysalicylate Aqueous Solution

[0163] 1.3 Experimental Results and Analysis

[0164] Experimental results analysis method:

[0165] Sample (sample group): In a 96-well plate, each group has 3 wells, and 100uL sample solution + 100uL DPPPH alcohol solution is added to each well;

[0166] Blank group: sample solution 100uL + anhydrous ethanol 100uL;

[0167] Control group: 100uL water + 100uL anhydrous ethanol.

[0168] The plate was placed at room temperature and protected from light for 30 minutes, and then the absorbance at 517 nm was measured using a spectrophotometer, and the average value was taken to calculate the DPPH clearance rate of each group of samples using the following formula.

[0169] DPPPH clearance rate = [1-(Asample-Ablank) / Acontrol] * 100%

[0170] Among them, Asample is the average absorbance value of the sample group;

[0171] Ablank is the absorbance of the blank group;

[0172] Acontrol is the absorbance of the control group.

[0173] Table 3 Experimental results

[0174]

[0175] It can be seen from the above experimental results that carnosine, ascorbyl glucoside and potassium 4-methoxysalicylate all have certain antioxidant effects. The DPPH scavenging rate of carnosine is 20.18%, the DPPH scavenging rate of ascorbyl glucoside is 16.52%, and the DPPH scavenging rate of potassium 4-methoxysalicylate is 14.56%. The DPPH scavenging rate of the combination of the three raw materials is 41.12%, which is significantly improved compared with the antioxidant effect of any single raw material, and has good antioxidant effect.

[0176] 2 Anti-glycation experiment

[0177] 2.1 Experimental Principle

[0178] Glycation refers to the combination of a carbohydrate molecule with other molecules such as proteins, lipids or DNA. Glycation is divided into enzymatic glycation and non-enzymatic glycation. Enzymatic glycation usually refers to glycosylation, such as the formation of a glycosidic bond in a sugar nucleotide molecule during the production of glycoproteins, while non-enzymatic glycation is what we call glycation in the skin, which refers to the direct chemical reaction between reducing sugars and proteins. This reaction causes the protein to cross-link and produce advanced glycation end products, commonly known as AGEs (Advanced Glycation Endproducts). The dark brown AGEs accumulate on the surface of the skin and cause the skin to become dull and yellow.

[0179] Advanced glycation end products (AGEs) are the end products of non-enzymatic glycosylation reactions (Maillard reactions). They refer to stable covalent adducts generated by the spontaneous reaction of macromolecules such as proteins, lipids or nucleic acids with glucose or other reducing monosaccharides without the participation of enzymes. AGEs have two sources in the body. One is the synthesis of excess sugar and protein in the body, and the other is the intake of AGEs in food through eating. The impact of AGEs on the skin is mainly reflected in the fact that it can cause skin aging and dull complexion. This experiment used a photoaging model in a high-sugar culture medium to explore the anti-AGES ability of carnosine.

[0180] Based on the principle of glycation reaction, relevant methods can be summarized: on the one hand, in the first step of glycation reaction, a substance that can replace the binding of protein and sugar molecules is found to form competitive inhibition, thereby preventing the protein from being glycosylated to form glycated protein. On the other hand, the glycated protein that has been glycosylated is deglycosylated to prevent the glycated protein from further oxidation and cross-linking, thereby reducing the production of glycation end products AGEs.

[0181] 2.2 Experimental methods

[0182] a) Cell culture: First, HSF cells (human skin fibroblasts) are cultured in DMEM high-glucose medium containing 10% fetal bovine serum to a suitable density, usually at 37°C and 5% CO2. It is recommended to replace fresh medium 2-3 times a week for cell culture, and use trypsin digestion for 1-2 minutes during subculturing, and subculture at a ratio of 1:2 to 1:4;

[0183] Take two bottles of HSF cells that have been cultured well, add 0.25% trypsin digestion solution, digest to make the attached cells fall off, count 1-4x 10^6 cells / mL, and make a cell suspension;

[0184] b) Establishment of UV light aging model: 100,000 cells / well of the cell suspension were appropriately diluted and inoculated on a 6-well plate. When the cells grew to about 80%, a UV light aging model was established.

[0185] Grouped as follows

[0186] Table 4

[0187] Serial number Experimental Group concentration 1 Blank control group + no UV irradiation / 2 Blank control group + UV irradiation / 3 DMEM high glucose medium containing 10% fetal bovine serum / 4 Skin biorhythm regulating factor + UV irradiation in the ratio of Example 1 0.5％ 5 Carnosine 100PPM 6 Skin biorhythm regulator factor + carnosine in the ratio of Example 1 0.5%+100PPM

[0188] After repeated washing with 1000ml PBS until colorless, 800μL PBS was added to the experimental group and irradiated under 80J / cm UV light, with the distance between the light source and the culture bottle being 15cm. After irradiation, the experimental group discarded PBS, added culture medium to dilute to 1200μL, and continued to incubate in a 37℃, 5% CO2 incubator for 48h;

[0189] c) scraping off the cells with a cell scraper, mixing by pipetting, ultrasonically disrupting the cell suspension for 30 seconds, and centrifuging at 10,000 g for 15 minutes to collect the cell supernatant to obtain the sample solution;

[0190] d) Follow the instructions of the Advanced Glycation End Product Detection Kit to detect its content.

[0191] 2.3 Experimental Results

[0192] Table 5 AGEs content

[0193]

[0194]

[0195] As shown in Table 5, compared with the blank control group, the AGEs content increased after UV radiation, and the AGES generation inhibition rates of adding 0.5% skin biorhythm regulator and 100PPM carnosine were 3.85% and 14.1%, respectively, indicating that 100ppm carnosine can significantly reduce the AGES content after UV radiation and has anti-glycation effect, while the skin biorhythm regulator has no obvious anti-glycation effect. After the combination of the two, the anti-glycation ability increased significantly, and the inhibition rate reached 50%, which was 35.9% higher than that of single carnosine, and the two had a synergistic effect. This may be related to the fact that the glycoproteins and amino acids in the biorhythm regulator have the function of protecting the protein structure in the cells.

[0196] 3 Anti-inflammatory experiments

[0197] 3.1 Experimental Principle

[0198] Inflammatory response is a biological response of the body to harmful stimuli such as pathogens, damaged cells or UV radiation. The inflammatory pathway consists of inducers, sensors, mediators and target tissues: various stimuli act on the skin as inducers, and Toll-like receptors (TLRs) and other sensors are expressed on sentinel cells such as macrophages, dendritic cells and mast cells, which sense and detect inducers. Sentinel cells and skin cells release mediators such as cytokines, chemokines, bioactive amines, eicosanoids and proteolytic cascade products (bradykinin). Inflammatory mediators act on various target tissues such as blood vessels and tissues, causing changes in their functional state and discomfort symptoms such as redness, swelling, heat and pain. Inflammatory response will lead to the production of inflammatory factors, which act on melanocyte receptors to produce melanin; at the same time, inflammatory response will promote the production of endothelin and stimulate melanocytes to produce melanin.

[0199] Tumor necrosis factor-α (TNF-α) is an inflammatory cytokine found in the tumor inflammatory microenvironment. It is mainly secreted by monocytes / macrophages and can initiate chronic inflammatory responses. TNF-α regulates inflammatory responses by activating two inflammatory signaling pathways, nuclear transcription factor KB (NF-KB) and activated protein 1 (AP-1). It is an important inflammatory factor and is involved in the pathological damage of certain autoimmune diseases. During the process of skin inflammation, TNF-α inflammatory factors are also secreted. When the inflammatory effect slows down, the inflammatory factors will also decrease.

[0200] There are the following anti-inflammatory methods for the inflammatory response pathway: first, reduce stimulation to avoid initiating an inflammatory response; block inflammatory signals to prevent cells from sensing and detecting inducers; inhibit macrophage activation and mast cell degranulation to reduce inflammatory mediators; regulate the secretion of inflammatory factors and reduce the release of cytokines such as TNF-α.

[0201] The basic principle of ELISA is to fix a certain concentration of antigen or antibody on the surface of a polystyrene microplate by physical adsorption, add the sample to be tested, and indirectly reflect the presence or amount of the antigen or antibody to be tested by the color depth of the enzyme label. Therefore, the anti-inflammatory effect of the test substance can be reflected by detecting the effect of the test substance on the production of TNF-α through ELISA.

[0202] Optical density (OD) is a parameter that measures the intensity of a substance's absorption of light of a specific wavelength in a solution. The optical density value is correlated with the concentration of the light-absorbing substance in the solution (for ELISA kits using the competitive method, the optical density value is negatively correlated with the concentration of the light-absorbing substance in the solution; for ELISA kits using the sandwich method, the optical density value is positively correlated with the concentration of the light-absorbing substance in the solution).

[0203] Bacterial lipopolysaccharide (LPS) binds to the antigen recognition receptors on the surface of macrophages, inducing macrophages to secrete multiple cytokines such as TNF-α. TNF-α can activate three signaling pathways, namely Caspase, JNK and transcription factor NF-kB, to achieve its biological functions such as cytotoxicity, antiviral, immunomodulation and apoptosis, and is one of the important inflammatory factors. LPS-induced RAW264.7 is a classic cell model for studying inflammatory factors. By comparing the difference in the secretion of TNF-a by RAW264.7 after administration of the negative control and the test substance, the effect of the test substance on inhibiting the secretion of TNF-a was evaluated.

[0204] The determination of TNF-a content adopts enzyme-linked immunosorbent assay (ELISA), and the specific principle is: TNF-a specifically binds to TNF-a antibody coated on the ELISA plate, and then binds to anti-TNF-a antibody with substrate labeling. After the substrate is catalyzed by the enzyme, a colored product is generated. The TNF-a content is positively correlated with the color depth of the colored product. The OD value is measured at a wavelength of 450nm using the enzyme label to calculate the TNF-a content.

[0205] 3.2 Experimental procedures

[0206] ① Experimental design

[0207] Table 6 Experimental design

[0208]

[0209]

[0210] ② Experimental methods

[0211] (1) Cell digestion and inoculation

[0212] Use trypsin / EDTA. The specific digestion concentration and dosage should be determined according to the characteristics of laboratory cells: the trypsin concentration should be 0.25%, the trypsin dosage for T25 culture flask is 1mL, and the trypsin dosage for T75 culture flask is 3mL.

[0213] Under microscope, when most of the cells become round and suspended, add DMEM medium containing serum at a volume of about 2 to 3 times that of trypsin to terminate digestion, collect the cells into a centrifuge tube, and centrifuge at 800 rpm for 6 min. The speed and centrifugation time can be determined according to the characteristics of the cells in the laboratory.

[0214] After centrifugation, discard the supernatant, add a certain volume of DMEM medium containing serum to the centrifuge tube, mix the cells by pipetting with an elbow pipette, and count the cells with a cell counter or a hemocytometer.

[0215] The cells were diluted with cell culture medium to the inoculation density (the degree of confluence reached 45% to 60% 24 hours after inoculation), and inoculated into a 96-well plate with a liquid volume of 200uL per well.

[0216] After inoculation, place in a CO2 incubator and culture for 24h±2h.

[0217] (2) Induction and administration

[0218] The culture medium in the 96-well plate was discarded, and the induction and drug administration operations were carried out.

[0219] The test substance concentration of experimental groups 1-5 at 100 ng / ml and culture medium containing 100 ng / ml LPS were added to the test substance wells;

[0220] The positive control wells need to be added with culture medium containing 100 ng / ml LPS;

[0221] DMEM medium containing serum was added to the blank / solvent control wells;

[0222] 200 μL per well.

[0223] After administration, the 96-well plate was placed in a CO2 incubator for 24 h±2 h.

[0224] After the incubation, 200 μL of cell culture supernatant was collected into a 1.5 mL sterile heart tube and stored in a -80°C ultra-low temperature refrigerator.

[0225] (3) ELISA test

[0226] ELISA testing should be performed according to the operating instructions of the TNF-a ELISA test kit.

[0227] Because the expression level of TNF-a is high after LPS induction, before the formal test, the dilution ratio should be set and a preliminary experiment should be conducted to determine the dilution ratio of the ELISA test group to ensure that the ELISA test value falls within the range of the standard curve.

[0228] The calculation formula of TNF-a inhibition rate is:

[0229] Inhibition rate (%) = (1-T / C) x 100%

[0230] Where:

[0231] T--average value of TNF-α content of the tested substances;

[0232] C- Average value of TNF-a content in negative control.

[0233] 3.3 Experimental Results and Analysis

[0234] The test results are as follows:

[0235] Table 7 Secretion of inflammatory factor TNF-a

[0236] Grouping Positive Control Experimental Group 1 Experimental Group 2 Experimental Group 3 Experimental Group 4 Experimental Group 5 TNF-α inhibition rate 0.01％ 25.71％ 50.25％ 2.79％ 87.37％ 107.67％ Significance analysis (P<0.05) Not significant Significant Significant Not significant Significant Significant

[0237] From the above results, it can be seen that the positive control group has no anti-inflammatory ingredients added, so there is no anti-inflammatory effect. Both citrus fruit extract and kava extract have significant anti-inflammatory effects, and carnosine has no significant effect. Compared with the negative control group, the TNF-α content of the kava extract group decreased by 25.71%, and the inflammatory factors in the citrus fruit extract group also decreased by 34.24%, both showing significant differences. Compared with the single raw material group, the inhibition rate of the composite sample of experimental group 4 reached 87.73%; the inhibition rate of the composite sample of experimental group 5 reached 107.67%, and the experimental group 5 was significantly improved by 20.3% relative to the experimental group 4, which may be due to the synergistic effect of carnosine on the inhibition of inflammatory factors. From the experimental results, it can be seen that kava extract and citrus fruit extract have anti-inflammatory effects, especially the combination of kava extract + citrus peel extract + carnosine has significant anti-inflammatory and soothing effects, and has a synergistic effect.

[0238] 4 Tyrosinase inhibition biochemical test

[0239] 4.1 Experimental Principle

[0240] In the biosynthesis of skin melanin, tyrosinase is a key enzyme that acts on DOPA to form DOPA quinone, which spontaneously undergoes a series of reactions to finally form melanin. In a phosphoric acid solution at pH 6.8, tyrosinase can catalyze the conversion of DOPA into DOPA quinone, and the absorbance value can be measured at 475nm on a spectrophotometer. Raw materials with tyrosinase activity inhibition can reduce the conversion of DOPA into DOPA quinone, thereby reducing the absorbance value. The inhibitory effect of the raw materials on tyrosinase activity can be evaluated based on the change in absorbance value.

[0241] 4.2 Experimental methods

[0242] 4.2.1 Preparation of experimental group solutions

[0243] Table 8 Solution configuration of experimental group

[0244]

[0245] 4.2.2 Experimental operation

[0246] (1) Preparation of phosphate buffered saline (PBS): Take 250 mL of 0.2 mol / L potassium dihydrogen phosphate solution, add 118 mL of 0.2 mol / L sodium hydroxide solution, and dilute to 1000 mL with deionized water (or directly weigh 6.8045 g of potassium dihydrogen phosphate and 0.944 g of sodium hydroxide, dissolve in deionized water and dilute to 1000 mL)

[0247] (2) L-tyrosine solution: weigh 27.17 mg L-tyrosine and dilute to 100 mL volumetric flask with PBS solution to prepare 1.5 mmol / L L-tyrosine solution, which is used immediately after preparation; (Determination of monophenolase activity)

[0248] (3) Preparation of tyrosinase solution: 25KU / 18.5mg tyrosinase (Solybol) was diluted to 2KU / mL by adding 12.5mL of the above PBS. The solution was stored in aliquots at -20°C for later use. When needed, the solution was diluted to 200U / mL with the above PBS.

[0249] (4) Determination of tyrosinase activity: Use a pipette to add solutions A, B, and C according to the dosage shown in the table below to a 96-well plate, and finally solution D. Mix thoroughly and incubate at 37°C for 10 min. Measure the absorbance A of the reaction solution at 475 nm and calculate its inhibition rate on tyrosinase.

[0250] Table 9

[0251] Tyrosine(A) Sample (B) Buffer (C) Tyrosinase (D) A0 50ul \ 100ul 50ul A1 50ul \ 150ul \ Sample A 50ul 50ul 50ul 50ul ABlank 50ul 50ul 100ul \

[0252] In Table 9, A0 represents the positive control group, A1 represents the blank group, A sample represents the experimental group, and A blank represents the blank (without tyrosinase) control group of the experimental group.

[0253] 5) Calculation formula:

[0254] Inhibition rate = [1-(A sample-A blank) / (A0-A1)] × 100%

[0255] 4.3 Experimental results and analysis

[0256] Table 10 Experimental results

[0257]

[0258]

[0259] It can be seen from the above results that the single ascorbyl glucoside and European thyme extract raw materials do not have a good effect on the inhibition of tyrosinase, and the tyrosinase inhibition rate is negative, which is not conducive to the whitening effect of the overall composition. The aqueous solution of carnosine + kava extract + citrus fruit extract + skin biorhythm regulator has the effect of inhibiting tyrosinase, but it is only 0.49%. The 4-methoxysalicylic acid potassium aqueous solution has a better effect of inhibiting tyrosinase, reaching 58.53%. The aqueous solution of ascorbyl glucoside + 4-methoxysalicylic acid potassium + European thyme extract has an effect of 65.78%. Therefore, the composition of the common raw materials has excellent effect, and the tyrosinase inhibition rate reaches 82.34%, indicating that the composition has whitening effect, and is better than the three whitening agents of ascorbyl glucoside + 4-methoxysalicylic acid potassium + European thyme alone.

[0260] 5. Cell Melanin Inhibition Experiment

[0261] 5.1 Experimental Principle

[0262] In this experiment, a B16-F10 melanocyte model was established by in vitro culture, kojic acid (which can inhibit cellular melanin synthesis) was used as a positive control, and the effect of the active ingredient (composition) on the intracellular melanin content was detected to reflect the whitening effect of the composition.

[0263] 5.2 Experimental methods

[0264] 1. Cell inoculation: B16-F10 cells were inoculated into 6-well plates and cultured in a 37°C, 5% CO2 incubator for 24 h;

[0265] 2. Dilution and addition of samples to be tested: Use DMDE high-glucose complete medium (DMEM basic medium, plus 10% FBS and 1% penicillin / streptomycin (P / S) antibiotics) as the diluent to prepare sample working solutions according to Table 11 for experimental groups 1 to 6, add cells, add an equal amount of complete medium to the negative control group, and add 100 μg / mL kojic acid to the positive control, and continue to culture in a 37°C, 5% CO2 incubator;

[0266] 3. Determination of intracellular melanin content: discard the supernatant, rinse, digest and collect the cells, centrifuge and remove the supernatant. Add 1M NaOH (10% DMSO) to resuspend the cell pellet, incubate at 80℃ for 2h, transfer the supernatant to a 96-well plate, add an equal amount of 1M NaOH (10% DMSO) to the blank control wells, and measure the absorbance of each well at a wavelength of 405nm;

[0267] 4. Calculation: After subtracting the background absorption from the absorbance value of each group, the absorbance value of the negative control group is taken as 100%, and the relative content of melanin in the cells of each concentration group is calculated. The calculation formula is as follows:

[0268] Relative content of intracellular melanin (%) = OD value of sample group / OD value of negative control group × 100%;

[0269] 5. Statistical analysis: GraphPad Prism 8.0 software was used for data statistical analysis and drawing charts. The quantitative data were expressed as x±s. The differences between the groups were analyzed by one-way ANOVA. P < 0.05 was considered statistically significant.

[0270] 5.3 Experimental Results and Analysis

[0271] Table 11

[0272]

[0273] It can be seen from the above results that the single ascorbyl glucoside, potassium 4-methoxysalicylate aqueous solution and European thyme extract all have a certain inhibitory effect on the synthesis of melanin. The aqueous solution of carnosine + kava extract + citrus fruit extract + skin biorhythm regulator has the worst effect of inhibiting melanin synthesis, which is only 1.9%. The aqueous solution of ascorbyl glucoside + potassium 4-methoxysalicylate + European thyme extract + carnosine + kava extract + citrus fruit extract + skin biorhythm regulator has the best effect, reaching 98.5%; the aqueous solution of ascorbyl glucoside + potassium 4-methoxysalicylate + European thyme extract is second, which is 69.4%. The above data show that the aqueous solution of ascorbyl glucoside + potassium 4-methoxysalicylate + thyme extract + carnosine + kava extract + citrus fruit extract + skin biorhythm regulator has a whitening effect, and is better than using ascorbyl glucoside, potassium 4-methoxysalicylate and thyme alone.

[0274] 6 Zebrafish melanin inhibition experiment

[0275] 6.1 Experimental Principle

[0276] Melanin in the human body is formed by the oxidation of tyrosine to dopa through the action of tyrosinase, which is then converted into indolequinone through oxidation and decarboxylation, and finally indolequinone is polymerized into melanin. The genes related to pigment formation in zebrafish are highly similar to those in humans in terms of sequence and regulatory mechanism. Zebrafish embryos are small and transparent, which makes it easy to take pictures under a microscope, and use software analysis to quantitatively read the melanin content level. The changes in the melanin levels of fish embryos in the treatment group and the control group are tested and compared, and the melanin inhibition rate is calculated to evaluate the melanin inhibition effect of the raw materials or products.

[0277] 6.2 Experimental methods

[0278] 6.2.1 System and sample size:

[0279] Fish strain used: wild-type AB strain zebrafish.

[0280] Zebrafish age: 6 hours post fertilization (6hpf).

[0281] The sample size of each experimental group was 15 (N=10).

[0282] Adult fish rearing and breeding methods: in line with the requirements of the international AAALAC certification (certification number: 001458).

[0283] 6.2.2 Test steps:

[0284] 1. Prepare Holt-Buffer mother solution, and use Holt-Buffer culture solution to dilute the sample to be tested 100 times to the experimental group concentration. Add the concentration of the sample to be tested in Table 12 to the selected zebrafish embryos, 200 μL per well, set more than 10 duplicate wells for each concentration, and add 1 zebrafish to each well. At the same time, set up a blank control group (Holt-Buffer culture solution group) and a positive control group (culture solution is 100mM arbutin solution), place it at 28.5±0.5℃ constant temperature culture for 48h, and change the solution every 24h.

[0285] Preparation method of Holt-Buffer culture medium: Accurately weigh 7.0g of sodium chloride, 0.4g of sodium bicarbonate, 0.1g of potassium chloride, and 0.235g of anhydrous calcium chloride, add to 900mL of deionized water, stir to dissolve, then make up to 2L with deionized water, filter and sterilize with a 0.20um filter, and store at room temperature.

[0286] 2. After the treatment, observe the pigmentation state of the embryos under a microscope and take pictures. Use Image J software to analyze the gray value of the pictures and calculate the relative melanin content.

[0287] 3. Data processing: According to the calculation formula:

[0288] Relative melanin content = average melanin content of sample group / average melanin content of blank control group × 100%.

[0289] 4. GraphPad Prism 8.0 software was used for data statistical analysis and graphs were drawn. The differences between the groups were analyzed using one-way ANOVA, and the differences were considered statistically significant when P < 0.05.

[0290] 6.3 Experimental Results and Analysis

[0291] The experimental results are shown in the following table:

[0292] Table 12

[0293]

[0294] From the above results, it can be seen that the melanin inhibition rate of the simple ascorbyl glucoside aqueous solution, the 4-methoxysalicylic acid potassium aqueous solution, and the European thyme extract aqueous solution is low; the melanin inhibition rate of the aqueous solution of carnosine + kava extract + citrus fruit extract + skin biorhythm regulator is the lowest, only 3.79%. The aqueous solution of ascorbyl glucoside + potassium 4-methoxysalicylic acid + European thyme extract + carnosine + kava extract + citrus fruit extract + skin biorhythm regulator has the best effect, reaching 37.36%. The above data show that the aqueous solution of ascorbyl glucoside + potassium 4-methoxysalicylic acid + European thyme extract + carnosine + kava extract + citrus fruit extract + skin biorhythm regulator has a whitening effect, and is better than the three whitening agents of ascorbyl glucoside, potassium 4-methoxysalicylic acid, and European thyme alone.

[0295] Examples 4 to 6 - Products

[0296] 1. Formulation

[0297] Table 13

[0298]

[0299] In Table 1, preservative A is p-hydroxyacetophenone, and preservative B is 1,2-hexanediol.

[0300] 2. Preparation method:

[0301] 1. Weigh the raw materials No. 1-4, add them into the main pot in sequence, heat to 80℃-85℃, keep warm for 30 minutes, then stir at 200r / min to make the BASE;

[0302] 2. Cool down to 40℃-45℃ and add No. 5 raw materials in sequence;

[0303] 3. Stir evenly, filter with filter cloth and discharge the material.

[0304] Test Example 2

[0305] 1 Safety Test of Products in Examples 4 to 6

[0306] 1 Materials and Methods

[0307] 1. Test substance: the original cosmetic final product.

[0308] 2. Negative control: blank control.

[0309] 3. Subjects: A total of 30 subjects, including 4 males and 26 females, aged 21 to 49 years old, with an average age of 30±7.7 years old, met the voluntary inclusion criteria.

[0310] 4. Patch test method: Select qualified patch test equipment, use the closed patch test method, place about 0.020mL to 0.025mL of the test substance of Example 4 in the patch tester, apply it to the curved side of the forearm of the subject with a hypoallergenic tape, remove the test substance after 24 hours, observe the skin reaction at 0.5, 24, and 48 hours after removal, and record the results according to the skin reaction grading standard in the "Cosmetic Safety Technical Specifications 2015". The scoring criteria are shown in Table 14, and the test results are shown in Table 15.

[0311] 2 Test results and conclusions

[0312] Table 14 Skin occlusion patch test skin reaction grading standard

[0313]

[0314]

[0315] Table 15 Test results

[0316]

[0317] Among them, the control group is an experimental group in which no experimental substances are added to the spot tester.

[0318] Test conclusion: No adverse reactions were observed in 33 people in the human skin patch test, indicating that the formula has good safety.

[0319] 2. Human efficacy test

[0320] 2.1 Soothing and repairing effect test

[0321] 2.1.1 Test Method

[0322] 33 Asian adult sensitive skin subjects aged 27 to 47 who often stay up late were screened and met the test criteria. They used Example 4 as an evaluation sample once in the morning and once in the evening for 28 consecutive days. The moisture content of the skin stratum corneum, the transepidermal water loss (TEWL) value, the skin hemoglobin content and the skin color were measured by an instrument probe on days 7, 14 and 28. Facial images were collected to analyze the skin color. The data were compared with the data before use to see if there was a significant difference, and the soothing and repairing effects were evaluated.

[0323] 2.1.2 Test results and conclusions

[0324] Table 16

[0325]

[0326]

[0327] See also Figure 1 After using the sample for 7 consecutive days, 96.97% of the subjects had a 9.56% increase in stratum corneum water content, a 6.79% decrease in TEWL value, a 3.21% decrease in skin hemoglobin content, and a 3.48% decrease in skin color (red-green) value a* value (CM-700d) of 36.9%. After using the sample for 14 consecutive days, 90.91% of the subjects had a 23.54% increase in stratum corneum water content, a 14.03% decrease in TEWL value, a 8.91% decrease in skin hemoglobin content. %, and the skin color (red-green) value a* value (CM-700d) decreased by 7.21%; after continuous use of the sample for 28 days, the water content of the stratum corneum of 93.94% of the subjects increased by 31.93%, the transepidermal water loss TEWL value decreased by 20.36%, the skin hemoglobin content decreased by 11.96%, and the skin color (red-green) value a* value (CM-700d) decreased by 7.68%, which were significantly different from those before use, indicating that the composition has short-term and long-term soothing and repairing effects.

[0328] 2 Whitening effect test

[0329] 2.1 Test Method

[0330] 33 Asian adult sensitive skin subjects aged 31 to 47 who often stay up late were screened and met the test criteria. After using the evaluation sample of Example 4 once in the morning and once in the evening for 28 consecutive days, the skin color L* value, (red-green) value a* value, (yellow-blue) value b* value, skin melanin content and skin color were measured by instrument probe, facial images were collected to analyze the spot optical density, and the ITA° value was calculated by the L* value, a* value, and b* value. At the same time, professional dermatologists evaluated and scored the skin color of the subjects before and after trying the test samples, and compared each dimension with the data before use to see whether there was a significant difference, and evaluated whether it had a whitening effect.

[0331] 2.2 Test results and conclusions

[0332] 2.2.1 Skin color L* value, (red-green) value a* value, (yellow-blue) value b* value and ITA° value

[0333] Table 17

[0334]

[0335]

[0336] 2.2.2 Skin melanin content, spot area ratio, and spot optical density

[0337] Table 18

[0338]

[0339] 2.2.3 Skin color assessment by dermatologists

[0340] Table 19

[0341]

[0342] 2.2.4 Conclusion

[0343] The samples were evaluated by 33 Asian adult subjects with sensitive skin aged 19 to 54 years old, and the results showed that the ITA° difference, MI difference and skin color visual score difference were significantly improved after 1 week, 2 weeks and 4 weeks of use compared with the negative control (P < 0.05). The regression analysis showed that the ITA° value, MI value and skin color visual score were significantly improved after 1 week, 2 weeks and 4 weeks of use compared with the negative control (P < 0.05). The combination has the effect of removing spots and whitening.

[0344] 3 Tests available for sensitive skin

[0345] 3.1 Test Method

[0346] Asian adult subjects with sensitive skin used the evaluation samples for 28 consecutive days under normal conditions. The dermatologist's safety evaluation and the subject's self-evaluation jointly determined whether it was "suitable for sensitive skin".

[0347] Safety assessment by dermatologists: Observe and evaluate the safety of samples on human skin through human trial tests. When the subjects return to the clinic, carefully inquire, check and record any adverse events that occurred during the use of the trial samples, including the discovery, time of occurrence, treatment measures and outcomes of the adverse events, and make a judgment on the relationship between the adverse events and the samples used.

[0348] Self-assessment by subjects: Subjects filled out a sample usage log based on 28 days of sample usage to record any adverse reactions to the skin caused by the samples; the evaluation time point was 1 time point in 28 days (28 days after using the samples).

[0349] 3.2 Test Results

[0350] Under the control of dermatologists, 33 Asian adult sensitive skin subjects aged 27 to 47 were observed to use the evaluation sample of Example 4 for 28 consecutive days under normal conditions to evaluate its potential to cause adverse reactions to human skin. The dermatologists' evaluation results showed that no adverse reactions were observed in the 33 subjects after using the sample, and the subjects' self-evaluation results showed that the 33 subjects did not feel uncomfortable during and after using the sample. The results meet the claim that the evaluation sample is "suitable for sensitive skin".

[0351] The above is only a preferred embodiment of the present invention. It should be pointed out that for ordinary technicians in this technical field, several improvements and modifications can be made without departing from the principle of the present invention. These improvements and modifications should also be regarded as the scope of protection of the present invention.

Claims

1. A whitening composition suitable for use on sensitive skin due to staying up late, characterized in that: include: 0.005 to 5 parts by weight of skin biorhythm regulating factor; Ascorbyl glucoside 0.02-3 parts by weight; 0.01 to 2 parts by mass of potassium 4-methoxysalicylate; 0.001 to 1 parts by weight of thyme extract; Carnosine 0.002-1 parts by weight; 0.001-1 parts by weight of Kava extract; 0.01 to 2 parts by mass of citrus fruit extract.

2. The whitening composition according to claim 1, characterized in that include: Skin biorhythm regulating factor 0.005 to 1 parts by weight; Ascorbyl glucoside 0.02-2 parts by mass; 0.01 to 1 parts by mass of potassium 4-methoxysalicylate; 0.001 to 1 parts by weight of thyme extract; Carnosine 0.002-0.2 parts by mass; 0.001-0.6 parts by weight of Kava extract; 0.01 to 1 part by weight of citrus fruit extract.

3. The whitening composition according to claim 1, characterized in that include: 0.5 parts by weight of skin biorhythm regulator; Ascorbyl glucoside 1.8 parts by mass; 0.8 parts by mass of potassium 4-methoxysalicylate; 1 part by weight of thyme extract; Carnosine 0.15 mass parts; 0.6 parts by weight of Kava extract; 1 part by weight of citrus fruit extract; Or, include: Skin biorhythm regulating factor 0.015 parts by mass; Ascorbyl glucoside 0.02 parts by mass; 0.02 parts by mass of potassium 4-methoxysalicylate; 0.01 parts by mass of thyme extract; Carnosine 0.015 parts by mass; 0.01 parts by weight of Kava extract; 0.01 parts by mass of citrus fruit extract; Or, include: Skin biorhythm regulating factor 0.005 parts by mass; Ascorbyl glucoside 0.02 parts by mass; 0.01 parts by mass of potassium 4-methoxysalicylate; 0.01 parts by mass of thyme extract; Carnosine 0.002 mass parts; 0.006 parts by weight of Kava extract; 0.01 parts by mass of citrus fruit extract.

4. The whitening composition according to claim 1, characterized in that The skin biorhythm regulating factor comprises a combination of yeast glycoprotein and amino acids, adenosine, and biorhythm peptides.

5. The whitening composition according to claim 1, characterized in that The skin biorhythm regulating factors include: 0.002 to 2 parts by weight of a combination of yeast glycoprotein and amino acids; 0.001 to 1 parts by weight of adenosine; 0.002 to 2 parts by mass of biorhythm peptide.

6. The whitening composition according to claim 1, characterized in that The kava extract is an extract of at least one of the leaves, roots and stems of kava.

7. Use of the whitening composition according to any one of claims 1 to 6 in whitening products.

8. A cosmetic, characterized in that: A whitening composition comprising any one of claims 1 to 6.

9. The cosmetic according to claim 6, characterized in that: The whitening composition is added in an amount of 0.0049% to 20% in cosmetics.

10. The cosmetic according to claim 6, characterized in that: include: 0.0049% to 20% whitening composition; 2% to 10% moisturizer; The balance is water.

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