Whitening application of cat's eye grass phenol D

Abstract

The present invention provides the whitening application of cat's eye phytophenol D. The present invention also relates to the application of cat's eye phytophenol D in the preparation of skin external preparations, medicines and health foods with whitening effects.

Description

Technical Field

[0001] The invention relates to the field of natural drug chemistry, and in particular to the whitening application of cat's eye grass phenol D, and the application of cat's eye grass phenol D in the preparation of a skin external preparation or medicine with whitening effect. Background Art

[0002] In recent years, as people's aesthetic level of skin appearance has improved, whitening cosmetics have been favored by more and more oriental women. Melanin deposition in human skin is regulated by multiple mechanisms, including transcription regulation, melanin synthesis regulation, melanin transport regulation, melanin distribution, degradation regulation, etc. At present, the main regulatory pathway of whitening preparations is to achieve whitening effects by inhibiting tyrosinase activity during melanin synthesis.

[0003] Melanin is a biological pigment produced in melanocytes. The deposition of melanin in the skin can cause skin dullness and other related problems. Inhibiting the production of melanin is an important way for the cosmetics industry to achieve whitening effects at this stage. In the process of melanin production, tyrosinase plays a major role. Tyrosinase is a type 3 multifunctional copper-containing glycoprotein located in the membrane of melanosomes and bound. Tyrosinase is only produced by melanocytes. After it is produced and processed in the endoplasmic reticulum and Golgi body, it is transported to the melanosomes and synthesized into melanin in the melanosomes. Tyrosinase is widely present in microorganisms, animals, plants and humans. It is an important rate-limiting enzyme involved in the melanogenesis process. Its expression level and activity determine the speed and output of melanin synthesis. Inhibiting the activity of tyrosinase can improve the tyrosinase metabolism of pigment cells in the skin, prevent the formation of pigmentation, and play a role in preventing the formation of freckles, chloasma, age spots, etc. and whitening. Therefore, regulating melanin synthesis by inhibiting tyrosinase is the main way to prevent pigmentation, and the pace of finding more effective tyrosinase inhibitors has never stopped.

[0004] At present, commonly used tyrosinase inhibitors mainly include kojic acid, hydroquinone, phenethylresorcinol and other resorcinol derivatives, which have good inhibitory ability on tyrosinase activity. Among them, phenethylresorcinol (i.e. 377) has been used as a common whitening ingredient in the field of skin and added to a variety of whitening skin care products. For example, CN201811150824.1 discloses a tyrosinase inhibitor, rutinone and rutinone, in a cream, a facial cleanser or a whitening lotion. Application, CN201811150810.X discloses a tyrosinase inhibitor, gingerol A, in a cream, a facial cleanser or a whitening lotion. Application, CN202110367742.8 discloses the application of snow lotus fermentation in inhibiting tyrosinase activity and melanin production, and CN202210382090.X discloses proanthocyanidins with the activity of inhibiting melanin production and its preparation method and use. There are also some patents for whitening compositions, involving purslane extract, β-glucan, arbutin, niacinamide, mulberry yellow, scutellaria extract, hesperidin, ginkgo leaf extract, rhodiola rosea extract, etc., but none of them involve whitening research on monomer compounds derived from Artemisia annua. It can be seen that the research on monomer compounds derived from Artemisia annua in the field of whitening is almost blank, and it may become a hot spot for exploration in the future.

[0005] Chrysosplenol D is also known as 3,6,7-trimethylquercetin, English name Chrysosplenol D, CAS No. 14965-20-9, molecular weight: 360.318, molecular formula: C 18 H 16 O8. The structural formula of cat's eye grass phenol D is:

[0006]

[0007] Catseye tannin D is a methoxylated flavonoid compound and the flavonoid compound with the highest content in Artemisia annua. In vitro activity shows that catseye tannin D has a selective synergistic effect on the antimalarial effect of artemisinin; in vitro studies have shown that catseye tannin D can induce cell cycle aberrations and apoptosis of breast cancer cells; in vivo studies have reported that catseye tannin D has a protective effect on LPS-induced systemic inflammatory response syndrome (SIRS) in mice. At present, most of the research on catseye tannin D is focused on the fields of anti-malaria, anti-inflammation and cell apoptosis, and there are few reports on its application in the field of skin, and no research on its whitening effect has been seen.

[0008] The present invention unexpectedly discovered that cat's eye grass phenol D can significantly inhibit the activity of tyrosinase, reduce the melanin content in melanocytes, and has a whitening effect. Summary of the invention

[0009] On the one hand, the present invention relates to the whitening application of cat's eye grass phenol D.

[0010] In a preferred embodiment, the concentration of cat's eye tannin D is at least 0.0001 mg / mL, preferably at least 0.001 mg / mL. In a preferred embodiment, the concentration of cat's eye tannin D is 0.001 mg / mL to 10 mg / mL.

[0011] In a preferred embodiment, the whitening is to inhibit tyrosinase activity, inhibit melanin production, inhibit spot production or a combination thereof.

[0012] On the other hand, the present invention provides the use of cat's eye grass phenol D in the preparation of a skin external preparation or medicine with whitening effect.

[0013] In a preferred embodiment, the content of cat's eye grass phenol D in the skin external preparation or medicine is 0.0001-20% by weight, preferably 0.001-1% by weight.

[0014] In a preferred embodiment, the skin external preparation is selected from: facial cream, lotion, gel, toner, essence, facial mask, eye cream, aerosol cleansing foam, spray, shower gel, or facial cleanser. DETAILED DESCRIPTION

[0015] The invention relates to the whitening effect of cat's eye phytophenol D. It is firstly discovered that cat's eye phytophenol D can be used as a functional additive to prepare a skin external preparation or medicine to achieve the whitening and brightening skin care effect.

[0016] In order to provide a more concise description, some quantitative expressions given herein are not modified by the term "about". It should be understood that, whether or not the term "about" is explicitly used, each quantity given herein is intended to refer to the actual given value, and is also intended to refer to the approximate values ​​of these given values ​​that can be reasonably inferred by ordinary technicians in the field, including the approximate values ​​of these given values ​​caused by experimental and / or measurement conditions.

[0017] To provide a more concise description, some quantitative expressions herein are described as a range of about X amount to about Y amount. It should be understood that when describing a range, the range is not limited to the upper and lower limits described, but should include the entire range of about X amount to about Y amount or any amount therebetween.

[0018] The cat's eye chloranol D described herein may optionally be in the form of a finished product package. In one embodiment, the package is a container such as a plastic, metal or glass tube or a wide-mouthed bottle containing cat's eye chloranol D. The product may additionally have a package such as a plastic or cardboard box for storing the container. In one embodiment, the product contains cat's eye chloranol D and has instructions for guiding the user to apply cat's eye chloranol D to the skin to treat the skin. Such instructions may be printed on the container, a label insert, or any other packaging.

[0019] As used herein, "topical application" means applying or spreading directly onto the external skin, scalp, or hair, for example, using the hands or an application device such as a wipe, roller, or spray.

[0020] As used herein, "cosmetically acceptable" means that the ingredients described by the term are suitable for use in contact with tissue (eg, skin or hair) without undue toxicity, incompatibility, instability, irritation, allergic response, or the like.

[0021] Catseye phytosan D

[0022] The present invention relates to the whitening effect of the compound cat's eye grass phenol D. In the study, it was found that cat's eye grass phenol D has an excellent whitening effect, can effectively inhibit the activity of tyrosinase, a key enzyme in melanin synthesis, and inhibit the production of melanin in melanocytes. Therefore, cat's eye grass phenol D can be used as an effective additive for preparing skin topical preparations or medicines to help improve whitening, brightening, and yellowing problems.

[0023] In whitening applications, the concentration of cat's eye tannin D is at least 0.0001 mg / mL, preferably at least 0.001 mg / mL. In a preferred embodiment, the concentration of cat's eye tannin D is 0.001 mg / mL to 10 mg / mL.

[0024] In some embodiments, the concentration of cat's eye tannin D is 0.001 mg / mL to 5 mg / mL. In some embodiments, the concentration of cat's eye tannin D is 0.001 mg / mL to 1 mg / mL, preferably 0.01 mg / mL to 1 mg / mL, more preferably 0.05 mg / mL to 1 mg / mL.

[0025] In some embodiments, the concentration of cat's eye tannin D is 0.018 mg / mL to 0.072 mg / mL. In other embodiments, the concentration of cat's eye tannin D is preferably 0.001125 mg / mL to 0.00225 mg / mL.

[0026] Skin external preparations or medicines containing physostigmol D

[0027] The cat's eye grass phenol D of the present invention can be used as a functional additive in skin external preparations or medicines.

[0028] The content of cat's eye grass phenol D in the skin external preparation or medicine is 0.0001-20% by weight, preferably 0.001-1% by weight.

[0029] In some embodiments, the drug is selected from the group consisting of tablets, capsules, emulsions, suspensions, powders, granules, solutions, and various pharmaceutical dosage forms known in the art. Different dosages are added according to different types of dosage forms.

[0030] In some embodiments, the skin topical agent is selected from the group consisting of: facial cream, lotion, gel, toner, essence, facial mask, eye cream, aerosol (cleansing foam), spray, shower gel and facial cleanser. Different amounts are added according to different types of preparations.

[0031] The skin external preparation is a general concept for all ingredients usually used on the outside of the skin, for example, it can be a cosmetic composition. The cosmetic composition can be a basic cosmetic, a facial makeup cosmetic, a body cosmetic, a hair care cosmetic, etc., and there is no special restriction on its dosage form, which can be reasonably selected according to different purposes. The cosmetic composition also contains different media or matrix excipients allowed by the cosmetics level according to different dosage forms and purposes.

[0032] The skin topical preparation comprising cat's eye grass phenol D can be topically applied to human skin and / or hair. The skin topical preparation may also include a cosmetically acceptable topical carrier, which may be about 50% by weight to about 99.99% by weight of the skin topical preparation (such as about 80% by weight to about 99% by weight of the skin topical preparation). In a preferred embodiment of the present invention, the cosmetically acceptable topical carrier includes water. The cosmetically acceptable topical carrier may include one or more selected from wetting agents, emollients, grease, wetting agents and similar substances. In one embodiment, the cosmetically acceptable topical carrier includes a substrate such as a nonwoven fabric or a membrane material.

[0033] Skin topical agents or medicines can be prepared into a variety of product types, including but not limited to lotions, creams, gels, sticks, sprays, ointments, clean liquid lotions and solid soaps, shampoos and hair conditioners, hair fixatives, pastes, foams, powders, mousse, shaving creams, wipes, patches, hydrogels, film-forming products, facial masks and skin films, films and cosmetics such as foundation and mascara. These product types can contain several types of cosmetically acceptable topical carriers, including but not limited to solutions, suspensions, emulsions (e.g., microemulsions and nanoemulsions), gels, solids and liposomes.

[0034] The skin topical preparation or medicine containing cat's eye phytosan D can be formulated as a solution. The solution usually contains an aqueous solvent or an organic solvent (e.g., about 50% to about 99.99% or about 90% to about 99% of a cosmetically acceptable aqueous solvent or organic solvent). Examples of suitable organic solvents include propylene glycol, polyethylene glycol, polypropylene glycol, glycerol, 1,2,4-butanetriol, sorbitol esters, 1,2,6-hexanetriol, ethanol, and mixtures thereof.

[0035] Skin topical preparations can be formulated into solutions containing emollients. Such skin topical preparations preferably contain from about 2% to about 50% of one or more emollients. As used herein, "emollient" refers to a substance used to prevent or alleviate dryness, for example, by preventing the loss of skin moisture through the skin. Examples of emollients include, but are not limited to, vegetable oils, mineral oils, aliphatic esters, etc.

[0036] Lotions can be prepared from such solutions.Lotions typically contain from about 1% to about 20% (e.g., from about 5% to about 10%) of one or more emollients and from about 50% to about 90% (e.g., from about 60% to about 80%) water.

[0037] Another type of product that can be formulated from a solution is a cream. Creams typically contain from about 5% to about 50% (e.g., from about 10% to about 20%) of one or more emollients and from about 45% to about 85% (e.g., from about 50% to about 75%) water.

[0038] Although it is preferred that the skin external preparation or medicine containing catsulphur D contains water, it may alternatively be anhydrous or an ointment containing no water but organic and / or silicone solvents, oils, lipids and waxes. The ointment may contain a simple base of animal or vegetable oil or semisolid hydrocarbon. The ointment may contain about 2% to about 10% of one or more emollients and about 0.1% to about 2% of one or more thickeners.

[0039] The topical agent for skin application can be formulated as an emulsion. If the topical carrier is an emulsion, about 1% to about 10% (e.g., about 2% to about 5%) of the topical carrier contains one or more emulsifiers. The emulsifier can be nonionic, anionic or cationic. Examples of suitable emulsifiers include those commonly identified as suitable emulsifiers in the field of personal care and cosmetic formulations.

[0040] Lotions and creams can be formulated as emulsions. Typically such lotions contain 0.5% to about 5% of one or more emulsifiers. Such creams typically contain about 1% to about 20% (e.g., about 5% to about 10%) of one or more emollients; about 20% to about 80% (e.g., 30% to about 70%) of water; and about 1% to about 10% (e.g., about 2% to about 5%) of one or more emulsifiers.

[0041] Oil-in-water and water-in-oil type single emulsion skin care preparations, such as lotions and creams, are well known in the cosmetics field and can be used in the present invention. Multiphase emulsion skin external preparations (such as water-in-oil-in-water type and oil-in-water-in-oil type) can also be used in the present invention. Generally, such single-phase emulsions or multiphase emulsions contain water, emollients and emulsifiers as their basic ingredients.

[0042] Skin topical preparations containing cat's eye grass phenol D can also be formulated into gels (e.g., aqueous gels, alcoholic gels, alcohol / water gels or oily gels using suitable gelling agents). Suitable gelling agents for aqueous gels and / or alcoholic gels include, but are not limited to, natural gums, acrylic acid and acrylate polymers and copolymers, and cellulose derivatives (e.g., hydroxymethyl cellulose and hydroxypropyl cellulose). Suitable gelling agents for oils (e.g., mineral oils) include, but are not limited to, hydrogenated butylene / ethylene / styrene copolymers and hydrogenated ethylene / propylene / styrene copolymers. Such gelling agents typically contain between about 0.1% and 5% by weight of such gelling agents.

[0043] The skin external preparation or medicine containing ophiopogon D can also be formulated into a solid preparation (eg, a wax-based stick, a bar soap, a powder, or a wipe containing the powder).

[0044] In addition to the above-mentioned components, the external preparation for skin which can be used in the present invention may contain various other oil-soluble substances and / or water-soluble substances at levels conventionally established in its technical field for use in external preparations for skin for use on skin and hair.

[0045] The skin topical preparation of the present invention may contain additional components commonly found in skin care compositions, such as moisturizers, skin regulators, emulsifiers, preservatives, antioxidants, fragrances, chelating agents, etc., as long as they are physically and chemically compatible with other components in the skin topical preparation and do not affect the effects of the cat's eye grass phenol D of the present invention.

[0046] In some embodiments of the skin topical preparation of the present invention, one or more preservatives may be used. Suitable preservatives include p-hydroxyacetophenone, C1-C4 alkyl parahydroxybenzoate and phenoxyethanol. Based on the total weight of the composition, the amount of the preservative is about 0.5 to about 2 weight %, preferably about 0.5 to 1 weight %.

[0047] In one example of the skin external preparation of the present invention, one or more antioxidants can be used. Suitable antioxidants include butylated hydroxytoluene (BHT), ascorbyl palmitate (BHA), butylated hydroxyanisole, phenyl-α-naphthylamine, hydroquinone, propyl gallate, nordihydroguaiaretic acid, vitamin E or vitamin E derivatives, vitamin C and its derivatives, calcium pantothenate, green tea extract and mixed polyphenols, and mixtures of the above-mentioned substances. The antioxidant used is about 0.02 to 0.5 weight % of the total weight of the composition, and more preferably from about 0.002 to 0.1 weight % of the amount used.

[0048] In one example of the skin external preparation of the present invention, one or more emollients can be used to act as lubricants through their ability to remain on the skin surface or in the stratum corneum to reduce flaking and improve the appearance of the skin. Typical emollients include fatty esters, fatty alcohols, mineral oils, polyether silicone copolymers, and the like. Examples of suitable emollients include, but are not limited to, polypropylene glycol ("PPG")-15 stearyl ether, PPG-10 cetyl ether, Steareth-10, Oleth-8, PPG-4 lauryl ether, vitamin E acetate, lanolin, cetyl alcohol, cetearyl ethylhexanoate, cetearyl alcohol, glyceryl stearate, hydroxystearate octyl, dimethyl polysiloxane, and combinations thereof. Cetyl alcohol, cetearyl ethylhexanoate, cetearyl alcohol, glyceryl stearate, and combinations thereof are preferred. When used, the emollient is used in an amount ranging from about 0.1 to about 30% by weight, preferably about 1 to about 30% by weight, based on the total weight of the composition.

[0049] In one example of the skin external preparation of the present invention, one or more moisturizers can be used. Moisturizers are also called humectants, which help to improve the effect of emollients, reduce flaking, remove the scales of the composition and improve the touch of the skin. Polyols can be used as moisturizers, including but not limited to glycerol, polyalkylene glycols, alkylene polyols and derivatives thereof, including butylene glycol, propylene glycol, dipropylene glycol, polyglycerol, polyethylene glycol and derivatives thereof, sorbitol, hydroxypropyl sorbitol, hexylene glycol, 1,3-dibutylene glycol, 1,2,6-hexanetriol, ethoxylated glycerol, propoxylated glycerol, and combinations thereof. When used, the amount of moisturizer is about 0.1 to about 20% by weight, preferably about 1 to about 15% by weight, based on the total weight of the composition.

[0050] In an example of the skin external preparation of the present invention, one or more emulsifiers can be used. Emulsifiers can be used within the scope of effective stabilizing amount. Preferably, based on the total weight of the composition, the emulsifier is used in an amount of about 1.0 to about 10.0 weight %, more preferably about 3.0 to about 6.0 weight %. Any emulsifier compatible with the components in the composition can be used. Suitable emulsifiers include stearic acid, cetyl alcohol, glyceryl stearate, lecithin, stearyl alcohol, Steareth-2, Steareth-20, acrylic acid (ester) class / C10-30 alkyl alcohol acrylate cross-linked polymer, and combinations thereof.

[0051] In one example of the skin external preparation of the present invention, one or more pH regulators can be used. The pH regulator useful in the skin external preparation of the present invention includes tromethamine. When used, the amount of the pH regulator based on the total weight of the composition is about 0.1 to about 2 weight %, preferably about 0.1 to about 1 weight %.

[0052] In a specific embodiment of the present invention, the skin topical agent comprises acrylates / C10-30 alkyl acrylate crosspolymer, glycerin, p-hydroxyacetophenone, glyceryl stearate and lecithin, cetearyl alcohol, cetearyl ethylhexanoate, tromethamine or a combination thereof.

[0053] Additional cosmetic active agents

[0054] In some embodiments, the topical skin preparation may also include additional cosmetic active agents. As used herein, "cosmetic active agents" are compounds (such as synthetic compounds or compounds isolated from natural sources or natural extracts) that have cosmetic or therapeutic effects on skin or hair, including but not limited to anti-acne agents, oil control agents, antimicrobial agents, anti-inflammatory agents, antifungal agents, antiparasitic agents, topical analgesics, sunscreens, photoprotectants, antioxidants, keratolytic agents, surfactants, moisturizers, nutrients, vitamins, energy enhancers, antiperspirants, astringents, deodorants, curing agents, anti-hard cuticle agents, and agents for hair and / or skin conditioning.

[0055] In one embodiment, these cosmetic actives are selected from (but not limited to): hydroxy acids, benzoyl peroxide, D-panthenol, octyl methoxycinnamate, titanium dioxide, octyl salicylate, homosalate, avobenzone, carotenoids, free radical scavengers, spin traps, amines, retinoids such as retinol and retinyl palmitate, ceramides, polyunsaturated fatty acids, essential fatty acids, enzymes, enzyme inhibitors, minerals, hormones such as estrogen, steroids such as hydrocortisone, 2-dimethylaminoethanol, copper salts such as copper chloride, copper-containing peptides such as Cu:Gly-His-Lys, coenzyme Q10, peptides, amino acids such as proline, vitamins, lactobionic acid, acetyl-CoA, niacin, riboflavin, thiamine, ribose, electron transfer substances such as NADH and FADH2, and other plant extracts such as aloe, feverfew, oatmeal, and their derivatives and mixtures. The cosmetic active agent is generally present in an amount of about 0.001% to about 20%, for example about 0.005% to about 10%, such as about 0.01% to about 5%, by weight of the topical skin preparation of the present invention.

[0056] Examples of vitamins include, but are not limited to, vitamin A, vitamin B (e.g., vitamin B3, vitamin B5, and vitamin B12), vitamin C, vitamin K, and different forms of vitamin E (e.g., α, β, γ, or Δ tocopherol) or mixtures thereof and derivatives thereof.

[0057] Examples of hydroxy acids include, but are not limited to, glycolic acid, lactic acid, malic acid, salicylic acid, citric acid, and tartaric acid.

[0058] Examples of antioxidants include, but are not limited to, water-soluble antioxidants such as sulfhydryl compounds and their derivatives (e.g., sodium metabisulfite and N-acetyl-cysteine), lipoic acid and dihydrolipoic acid, resveratrol, lactoferrin and ascorbic acid and ascorbic acid derivatives (e.g., ascorbyl palmitate and ascorbic acid polypeptide). Oil-soluble antioxidants suitable for use in the skin topical preparations of the present invention include, but are not limited to, butylated hydroxytoluene, retinoids (e.g., retinol and retinyl palmitate), tocopherols (e.g., tocopherol ethyl ester), tocotrienols and ubiquinone. Natural extracts containing antioxidants suitable for use in the skin topical preparations of the present invention include, but are not limited to, extracts containing flavonoids and isoflavones and their derivatives (e.g., genistein and di-zein), extracts containing resveratrol, and the like. Examples of such natural extracts include grape seeds, green tea, pine bark, and propolis.

[0059] How to use

[0060] The skin topical preparation of the present invention can be topically applied to the skin of a mammal of one or more skins in need of treatment. In one embodiment, the skin topical preparation can be applied to the skin in need of whitening and brightening. The skin topical preparation can be applied to the skin in need of such treatment according to a suitable treatment regimen, such as monthly, weekly, every other day, daily, twice a day, etc.

[0061] In certain embodiments, the skin topical preparations of the present invention can also be used to treat other needs related to the skin. For example, the skin topical preparations of the present invention can be used to treat hyperpigmentation after inflammation, to reduce pore size, to reduce sebum production, and to alleviate scars. In certain other embodiments, the skin topical preparations of the present invention can be applied simultaneously with a mechanical or physical epidermal exfoliation treatment (e.g., microdermabrasion treatment) or within a few hours of the treatment, or simultaneously with a chemical peel or keratolytic agent such as salicylic acid. In certain other embodiments, the skin topical preparations of the present invention can be applied to mucous membranes or other tissues such as vaginal tissue, oral tissue, or eye tissue. In certain other embodiments, the skin topical preparations of the present invention can be applied to mild wounds or postoperative sites to promote healing, to insect bites, to poison ivy skin disorders or similar skin conditions, or generally used to relieve itching.

[0062] Example

[0063] The present invention is further described below in conjunction with specific examples. It is necessary to point out that the examples are only used to further illustrate the present invention and cannot be construed as limiting the scope of the present invention. Those skilled in the art can make some non-essential improvements and adjustments based on the content of the present invention described above. The test methods in the following examples that do not specify specific conditions are usually based on conventional conditions or the conditions recommended by the manufacturer. Unless otherwise stated, all percentages and parts are by weight.

[0064] The cat's eye phytophenol D used in the present invention has a purity of more than 90%, is derived from Artemisia annua, and is studied by the Chinese Academy of Chinese Medical Sciences.

[0065] Example 1: Preparation of cat's eye grass phenol D

[0066] Weigh 36.0 mg of cat's eye phytosan D, dilute to 100 mL volumetric flask with dimethyl sulfoxide (Sigma, DMSO) to obtain a 0.36 mg / mL cat's eye phytosan D solution for later use.

[0067] Example 2: Preparation of cat's eye grass phenol D

[0068] Measure 3 mL of the solution of Example 1 and dilute it to 4 mL with DMSO to obtain a 0.27 mg / mL cat's eye grass phenol solution for later use.

[0069] Example 3: Preparation of cat's eye grass phenol D

[0070] Measure 2 mL of the solution in Example 1 and dilute it to 4 mL with DMSO to obtain a 0.18 mg / mL cat's eye grass phenol solution for later use.

[0071] Example 4: Preparation of cat's eye grass phenol D

[0072] Measure 1 mL of the solution in Example 1 and dilute it to 4 mL with DMSO to obtain a 0.09 mg / mL cat's eye grass phenol solution for later use.

[0073] Example 5: Preparation of cat's eye grass phenol D

[0074] Weigh 18 mg of cat's eye tannin D, dilute to a 10 mL volumetric flask with DMSO to obtain a 1.8 mg / mL cat's eye tannin D solution for later use.

[0075] Example 6: Preparation of cat's eye grass phenol D

[0076] Measure 1 mL of the solution of Example 5, dilute it to 100 mL with DMSO to obtain a 0.018 mg / mL cat's eye grass phenol solution, and then measure 1 mL of the above solution, dilute it to 8 mL with DMSO to obtain a 0.00225 mg / mL cat's eye grass phenol solution for use.

[0077] Example 7: Preparation of cat's eye grass phenol D

[0078] Take 2 mL of the solution of Example 6 and dilute it to 4 mL with DMSO to obtain a 0.001125 mg / mL cat's eye grass phenol solution for later use.

[0079] Test Example 1: Tyrosinase activity assay

[0080] 1. Test method:

[0081] The tyrosinase activity test uses L-DOPA as a substrate. In the test, a sample group (T), a sample background group (T0), an enzyme reaction group (C) and a solvent control group (C0) were set up. The sample group was the vitexin solution of Examples 2, 3, 4, and 5. Use a 96-well plate with 200 μL wells to add liquid according to the table below. Three replicate wells need to be set up for each sample well (T) of each tested concentration of each sample, and three replicate wells need to be set up for the enzyme reaction well (C). After thorough mixing, place in a 37°C incubator and incubate for 10 minutes, and use an enzyme reader to detect the absorbance value of the sample at 492 nm. Calculate the tyrosinase inhibition rate:

[0082]

[0083] Table 1: Sample dosing requirements

[0084]

[0085]

[0086] 2. Test groups and concentrations:

[0087]

[0088] Note: Due to the influence of the test system, the concentration of the prepared test sample will be further diluted 5 times during the test, so the prepared concentration is 5 times the final test concentration. The concentration involved in the test result explanation also refers to the final test concentration.

[0089] 3. Test results:

[0090] Table 2: Tyrosinase activity inhibition rate results

[0091]

[0092] As can be seen from Table 2, the tyrosinase inhibition rate of cat's eye grass phenol D in Example 1 (0.072 mg / mL) is 82.17%, the tyrosinase inhibition rate of cat's eye grass phenol D in Example 2 (0.054 mg / mL) is 48.75%, the tyrosinase inhibition rate of cat's eye grass phenol D in Example 3 (0.036 mg / mL) is 31.63%, and the tyrosinase inhibition rate of cat's eye grass phenol D in Example 4 (0.018 mg / mL) is 23.12%.

[0093] The results showed that in the in vitro tyrosinase activity inhibition experiment, 0.018-0.072 mg / mL of cat's eye grass phenol D could effectively inhibit tyrosinase activity in a concentration-dependent manner.

[0094] Test Example 2: Test on the inhibition of melanin production in melanocytes by cat's eye grass phenol D

[0095] 1. Test Principle

[0096] The production of melanin by melanocytes in the skin is the main cause of skin darkening and the formation of spots. Therefore, this test uses melanocytes (primary melanocytes, Guangdong Boxi Biological) as the test tool to evaluate the whitening effect of the sample to be tested by detecting changes in melanin content.

[0097] 2. Test Method

[0098] Press 3×10 5 Cells were inoculated into 6-well plates at a seeding density of 100 cells / well and incubated overnight in an incubator (37°C, 5% CO2). When the cell plating rate in the 6-well plate reached 40% to 60%, the cells were divided and dosed, with 3 replicates per group. After incubation in the incubator for 72 hours, the cells were digested, melanin extract was added, melanin was extracted by heat incubation, and finally added to a 96-well plate to measure the OD value at 405nm.

[0099] 2.1 Main Reagents

[0100] Human melanocytes (Guangdong Boxi Biotechnology Co., Ltd.), M-254 culture medium (Boxi Biotechnology), MTT (Sigma), DMSO (Sigma), PBS (Solebo), glabridin (Shanghai Oli), melanin standard (Sigma), sodium hydroxide (Sinopharm Group), anhydrous ethanol (Sinopharm Group), ether (Sinopharm Group), and ammonia (Sinopharm Group).

[0101] 2.2 Main Equipment

[0102] CO2 incubator (Thermo, 150I), clean bench (Sujing Antai, SW-CJ-1F), microplate reader (BioTek, Epoch), digital constant temperature water bath (Changzhou Guohua, HH-4A).

[0103] 3. Test results

[0104] Table 3: Melanin production inhibition results

[0105]

[0106] Note: Inhibition rate (%) = (melanin content of control group - melanin content of sample) / melanin content of control group.

[0107] As shown in Table 3, Example 7 (0.00225 mg / mL cat's eye grass phenol D solution) can significantly inhibit the production of melanin in melanocytes, with an inhibition rate of 20.37%; Example 8 (0.001125 mg / mL cat's eye grass phenol D solution) can significantly inhibit the production of melanin in melanocytes, with an inhibition rate of 14.71%. This indicates that cat's eye grass phenol D can effectively inhibit the production of melanin in an in vitro melanocyte model at a concentration of 0.001125 to 0.00225 mg / mL, and the effect of 0.00225 mg / mL concentration is better.

[0108] The cat's eye grass phenol D of the present invention can be used as an intermediate raw material or an effective additive for the preparation of a skin topical preparation or a medicine. The skin topical preparation is preferably a cosmetic composition, including but not limited to the preparation of products in the form of cream, lotion, gel, lotion, essence, mask, eye cream, aerosol (cleansing bubble), spray, shower gel and facial cleanser. The weight percentage of cat's eye grass phenol D solution in the skin topical preparation is 0.0001%-20% (w / w). The preferred weight percentage is 0.001%-10% (w / w). The more preferred weight percentage is 0.001%-5% (w / w). The more preferred weight percentage is 0.001%-1%. The most preferred weight percentage is 0.005%-0.1% (w / w). In some embodiments, the weight percentage of cat's eye grass phenol D in the skin topical preparation is 0.0018%-0.0072% (w / w). The preferred weight percentages are 0.0036%, 0.0054%, and 0.0072% (w / w). The more preferred weight percentages are 0.0054% and 0.0072% (w / w). The most preferred weight percentage is 0.0072% (w / w).

[0109] The following are specific application examples of cat's eye grass phenol D solution in skin external preparations or medicines according to Examples 1-8, as well as the formulations and preparation methods of these dosage forms. In the following tables, "-" means no addition.

[0110] Application Example 1: Preparation of facial cream

[0111]

[0112] Application Example 2: Preparation of Emulsion

[0113]

[0114]

[0115] Application Example 3: Preparation of Gel

[0116]

[0117] Application Example 4: Preparation of lotion

[0118]

[0119] Application Example 5: Preparation of Essence

[0120]

[0121]

[0122] Application Example 6: Preparation of facial mask

[0123]

[0124] Application Example 7: Preparation of Eye Cream

[0125]

[0126] Application Example 8: Preparation of Aerosol (Cleaning Bubble)

[0127]

[0128] Application Example 9: Preparation of Spray

[0129]

[0130] Application Example 10: Preparation of Shower Gel

[0131]

[0132]

[0133] Application Example 11: Preparation of facial cleanser

[0134]

Claims

1. Non-therapeutic whitening application of cat's eye tannin D, wherein the concentration of cat's eye tannin D is 0.001 mg / mL to 1 mg / mL.

2. The use according to claim 1, characterized in that The use concentration of cat's eye grass phenol D is 0.001125 mg / mL to 0.36 mg / mL.

3. The use according to claim 1 or 2, characterized in that: The whitening is to inhibit tyrosinase activity.

4. The use according to claim 1 or 2, characterized in that: The whitening is to inhibit the production of melanin.

5. The use according to claim 1 or 2, characterized in that: The whitening is to inhibit the generation of spots.

6. Use of cat's eye grass phenol D in the preparation of a skin external preparation or medicine with whitening effect, wherein the concentration of cat's eye grass phenol D is 0.001 mg / mL to 1 mg / mL.

7. The use according to claim 6, characterized in that The content of cat's eye grass phenol D in the skin external preparation or medicine is 0.0001-20% by weight.

8. The use according to claim 7, characterized in that The content of cat's eye grass phenol D in the skin external preparation or medicine is 0.001-1% by weight.

9. The use according to claim 6, characterized in that The skin external preparation is selected from: facial cream, lotion, gel, toner, essence, facial mask, eye cream, aerosol cleansing foam, spray, shower gel, or facial cleanser.

10. The use according to any one of claims 6 to 9, characterized in that: Features The whitening is to inhibit tyrosinase activity.

11. The use according to any one of claims 6 to 9, characterized in that: Features The whitening is to inhibit the production of melanin.

12. The use according to any one of claims 6 to 9, characterized in that: The whitening is to inhibit the generation of spots.

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