Anti-saccharification application of osmundone

Abstract

The invention provides an anti-saccharification application of osmundone. The anti-saccharification of the present invention is an end product that inhibits non-enzymatic glycosylation reactions. The anti-saccharification of the invention may also be the inhibition of glycosylation end products. The invention also relates to application of osmundone in preparation of an external skin preparation, a medicine and a health food with an anti-saccharification effect. The external skin preparation is selected from face cream, emulsion, gel, toning lotion, essence, a mask, eye cream, aerosol cleaning foam, spray, shower gel or facial cleanser.

Description

Technical Field

[0001] The invention relates to the field of natural drug chemistry, and in particular to the anti-glycation application of porphyrin, and the application of porphyrin in the preparation of skin external preparations, medicines and health foods with anti-glycation effects. Background Art

[0002] As we age, our metabolism slows down, life and work pressures increase, and with irregular lifestyles and eating habits, the sugar we consume will not be metabolized immediately and will accumulate in the body. In the absence of enzymes, free sugars react with collagen in the dermis to form some reversible primary "glycosylation products", which then form highly fluorescent, cross-linked and irreversible brown non-enzymatic glycation end products (AGEs). The whole process is called non-enzymatic glycosylation (NEG), and AGEs can further directly cross-link biological macromolecules in the body, causing functional damage.

[0003] When elastin and collagen in the skin participate in non-enzymatic glycation reactions to generate AGEs, a cross-linked product of collagen and elastin is formed. As collagen cross-linking gradually increases, the solubility of collagen decreases, insoluble collagen accumulates, and the permeability of skin tissue decreases, resulting in barriers to the exchange of nutrients and metabolites between cells, which ultimately leads to decreased skin elasticity, sagging, and wrinkles. In addition, glycated collagen turns yellow and becomes brittle, so as we age, AGEs accumulate in large quantities and the skin gradually turns yellow. In addition, when the receptors of AGEs (AGER) are activated, they induce the body to produce a large number of free radicals, which adds a large burden to the antioxidant system in the body, further damages the skin tissue, stimulates the production of melanin, and puts the skin into a vicious cycle. Therefore, in recent years, the anti-glycation theory has become a hot mechanism and target for skin beauty. The anti-glycation effect is not only closely related to skin aging, but also closely related to the dark yellow color of the skin.

[0004] At present, the representative anti-glycation ingredients include aminoguanidine, carnosine, lipoic acid, niacinamide, quercetin and curcumin, etc. The mechanism is mainly to block the development of AGEs and inhibit the generation of AGEs. Among them, aminoguanidine is a drug known to effectively prevent the glycation process, and carnosine is a recognized effective anti-glycation ingredient in the skin field.

[0005] In recent years, natural Chinese herbal anti-glycation inhibitors are still a hot topic that people are pursuing. According to searches in recent years, Chinese patent application 201810568900.4 discloses an anti-glycation black tea extract and a preparation method thereof. Chinese patent application 202210159009.1 discloses the anti-glycation use and detection method of Bauhinia dahurica extract. Chinese patent application 202111648102.0 discloses a Chinese herbal extract anti-glycation. At the same time, there are many other patents for anti-glycation compositions, involving bamboo leaf lily, mangosteen, rhodiola rosea dandelion, etc. However, none of these documents involve the anti-glycation research of violaceous ketone.

[0006] Osmundacetone, Chinese name 3,4-dihydroxybenzylideneacetone, English name Osmundacetone, Cas number 123694-03-1 or 37079-84-8, molecular formula C 10 H 10 O3, exists in Osmanthus fragrans and Inonotus obliquus. The structural formula of Osmanthus fragrans is:

[0007] .

[0008] Modern synthesis routes basically use 3,4-dihydroxybenzaldehyde or 3,4-bis(methoxymethoxy)benzaldehyde as raw materials. Modern pharmacology shows that purpurogenol can regulate blood pressure and lower cholesterol.

[0009] For example, Chinese patent application 201780074399.X discloses a composition containing osfera ketone or a pharmaceutically acceptable salt thereof for preventing or treating bone diseases, and reports that Qi Zitong controls oil to improve symptoms of bone diseases and activates osteoblast differentiation. The patent also mentions that "the preferred content of the food composition of the present invention is 0.01-50wt%". At the same time, the patent application also shows that in all tests of human toxicity, irritation, carcinogenicity, environmental toxicity, etc. by many U.S. toxic substance regulatory agencies, purpurogenol was determined to be harmless and safe (Note: TSCA: not listed; CLEAN WATER ACT (CWA): not listed; SARA313: not listed; MARINE POLLUTANT: not listed; RIGHT TO KNOW LIST (NEW JERSEY): not listed; RIGHT TO KNOW LIST (MASSACHUSETTS): not listed; RIGHT TO KNOW LIST (PENNSYLVANIA): not listed; ILLINOISTOXIC AIR CONTAMINANTS: not listed; CLEAN AIR ACT (CAA): not listed; DHS CHEMICALS OF INTEREST: not listed; CALIFORNIA PROPOSITION 65: not listed). PROP65): Not listed; OSHA: Not listed; CALIFORNIAPROP 65TOXICITY TYPE(CANCER, DEVELOPMENTAL, FEMALE, MALE): None; OSHA HAZARD CLASS(CARCINOGEN, CORROSIVE, FLAMABLE, REACTIVE, TOXIC): None, etc.).

[0010] At present, no relevant reports on the application of porphyrin in cosmetics have been found in the literature. The present invention unexpectedly found that porphyrin has excellent anti-glycosylation effect and can effectively reduce the content of AGEs, the end product of non-enzymatic glycosylation reaction. Summary of the invention

[0011] The invention provides the anti-glycation application of porphyrone.

[0012] In a preferred embodiment, porphyrin is used at a concentration of at least 0.1 μg / mL, more preferably at least 0.5 μg / mL.

[0013] In a preferred embodiment, anti-glycation is to inhibit the end products of non-enzymatic glycosylation reactions. In a preferred embodiment, anti-glycation is to inhibit the end products of glycation.

[0014] The invention also relates to the application of porphyrin in the preparation of medicines, health foods or skin external preparations with anti-glycation effects.

[0015] In a preferred embodiment, the content of porphyrin in the medicine, health food, or external skin preparation is 0.00001-20% by weight, more preferably 0.0001-5% by weight.

[0016] 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.

[0017] BRIEF DESCRIPTION OF THE DRAWINGS

[0018] Figure 1 The results of the anti-glycation test of porphyrone are shown.

[0019] Figure 2A The fluorescence staining image of the anti-glycation efficacy of the blank group is shown. Upright fluorescence microscope Leica DM2500, magnification 200 times.

[0020] Figure 2B The negative control group is shown in fluorescence staining of anti-glycation efficacy. Upright fluorescence microscope Leica DM2500, magnification 200 times.

[0021] Figure 2C The anti-glycation efficacy fluorescence staining image of the positive control group is shown. Upright fluorescence microscope Leica DM2500, magnification 200 times.

[0022] Figure 2D The fluorescence staining diagram of the anti-glycation efficacy of the 0.178 μg / mL porphyrin test group is shown, where ZYS represents porphyrin. Upright fluorescence microscope Leica DM2500, magnification 200 times.

[0023] Figure 2E The fluorescence staining diagram of the anti-glycation efficacy of the 0.89 μg / mL porphyrin test group is shown, where ZYS represents porphyrin. Upright fluorescence microscope Leica DM2500, magnification 200 times. DETAILED DESCRIPTION

[0024] The present invention relates to the anti-glycation effect of osfera ketone, and for the first time finds that osfera ketone can be used as a functional additive to prepare a skin external preparation to achieve skin care effects such as whitening, brightening, yellowing, and anti-aging. In some embodiments, osfera ketone can be used as a functional additive to prepare medicines or health foods.

[0025] 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.

[0026] 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.

[0027] Osorbitrone described herein may optionally be in the form of finished product packaging. In one embodiment, the packaging is a container such as a plastic, metal or glass tube or a wide-mouthed bottle containing osorbitrone. The product may additionally have a packaging such as a plastic or cardboard box for storing the container. In one embodiment, the product comprises osorbitrone and has instructions for guiding the user to apply osorbitrone to the skin to treat the skin. Such instructions may be printed on the container, a label insert or any other packaging.

[0028] 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.

[0029] 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.

[0030] Porphyrone

[0031] The present invention relates to the anti-glycation effect of the compound porphyrin, and it is found in the study that porphyrin can effectively reduce the generation of AGEs, the end product of non-enzymatic glycation reaction, and has excellent anti-glycation effect. Therefore, porphyrin can be used as an effective additive for preparing medicines, health foods or skin topical preparations to achieve skin care effects such as whitening, brightening, yellowing, and anti-aging.

[0032] In anti-glycation applications, porphyrin is used at a concentration of at least 0.1 μg / mL, more preferably at least 0.5 μg / mL.

[0033] In some embodiments, osfera ketone is used at a concentration of 0.1-20 μg / mL. In some embodiments, osfera ketone is used at a concentration of 0.1-10 μg / mL, preferably 0.1-5 μg / mL, preferably 0.1-1 μg / mL. In a specific embodiment, osfera ketone is used at a concentration of 0.89 μg / mL.

[0034] Medicines, health foods or skin topical preparations containing porphyrin

[0035] The porphyrin of the present invention can be used as an effective additive in medicines, health foods or external skin preparations.

[0036] In some embodiments, the content of porphyrin in the medicine, health food or skin external preparation is 0.00001-20% by weight, more preferably 0.0001-5% by weight.

[0037] 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.

[0038] Health food is also called functional food. It has the function of regulating human body functions, but is not for the purpose of treating diseases, and is suitable for consumption by specific groups of people. In some embodiments, the health food can be in powder form.

[0039] 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.

[0040] Skin external preparations are a general concept for all ingredients that are usually applied to the outside of the skin, such as cosmetic compositions. The cosmetic compositions may include basic cosmetics, facial makeup cosmetics, body cosmetics, hair care cosmetics, etc., and there is no particular restriction on the dosage form, which can be reasonably selected according to different purposes. The cosmetic compositions may also contain different media or matrix excipients permitted by cosmetics according to different dosage forms and purposes.

[0041] The skin topical preparation comprising porphyrone can be topically applied to human skin and / or hair. The skin topical preparation can also include a cosmetically acceptable topical carrier, which can 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 comprises 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 comprises a substrate such as a nonwoven fabric or a membrane material.

[0042] Skin topical agents can be prepared into a variety of product types, including but not limited to lotions, creams, gels, sticks, sprays, ointments, cleansing liquid lotions and solid soaps, shampoos and hair conditioners, hair fixatives, pastes, foams, powders, mousses, 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.

[0043] The topical skin preparation containing porphyrin can be formulated as a solution. The solution generally 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.

[0044] 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.

[0045] 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.

[0046] 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.

[0047] Although it is preferred that the topical skin preparation containing porphyrin contains water, alternatively the topical skin preparation may be anhydrous or an ointment containing no water but organic and / or silicone solvents, greases, lipids and waxes. The ointment may contain a simple base of animal or vegetable oils or semisolid hydrocarbons. The ointment may contain from about 2% to about 10% of one or more emollients and from about 0.1% to about 2% of one or more thickeners.

[0048] 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.

[0049] 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.

[0050] 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.

[0051] The skin topical preparation comprising osfera ketone can also be formulated into a gel (e.g., an aqueous gel, an alcoholic gel, an alcohol / water-type gel or an oily gel using a suitable gelling agent). 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.

[0052] The external skin preparation containing porphyrin may also be formulated into a solid preparation (eg, a wax-based stick, a bar soap, a powder, or a wipe containing the powder).

[0053] 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.

[0054] The skin topical preparation of the present invention may contain additional components commonly found in skin care compositions, such as emollients, 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 porphyrin of the present invention.

[0055] 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 %.

[0056] 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.

[0057] 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.

[0058] 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.

[0059] 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.

[0060] 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 %.

[0061] 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.

[0062] Additional cosmetic active agents

[0063] 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.

[0064] 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.

[0065] 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.

[0066] Examples of hydroxy acids include, but are not limited to, glycolic acid, lactic acid, malic acid, salicylic acid, citric acid, and tartaric acid.

[0067] 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.

[0068] How to use

[0069] The skin preparation of the present invention can be topically applied to the skin of a mammal of one or more skins that need to be treated. In one embodiment, the skin preparation can be applied to the skin that needs to be treated for whitening, brightening, yellowing, and anti-aging. The skin preparation can be applied to the skin that needs such treatment according to a suitable treatment regimen, such as monthly, weekly, every other day, daily, twice a day, etc.

[0070] 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.

[0071] Example

[0072] 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.

[0073] The porphyrin used in the examples of the present invention was provided by the Institute of Chinese Materia Medica, China Academy of Chinese Medical Sciences.

[0074] Example 1: Preparation of porphyrone

[0075] Weigh 1.78 mg of porphyrin ketone and dilute it to a 10 mL volumetric flask with dimethyl sulfoxide (Sigma, DMSO) to obtain a 0.178 mg / mL porphyrin ketone solution for later use.

[0076] Example 2: Preparation of porphyrone

[0077] Weigh 8.9 mg of porphyrin ketone and dilute to 10 mL volumetric flask with DMSO to obtain 0.89 mg / mL porphyrin ketone solution for later use.

[0078] Example 3: Preparation of porphyrone

[0079] Weigh 5.34 mg of porphyrin ketone and dilute to 10 mL volumetric flask with DMSO to obtain 0.534 mg / mL porphyrin ketone solution for later use.

[0080] Example 4: Preparation of porphyrone

[0081] Weigh 17.8 mg of porphyrin ketone and dilute to 10 mL volumetric flask with DMSO to obtain a 1.78 mg / mL porphyrin ketone solution for later use.

[0082] Example 5: Evaluation of the anti-glycation properties of porphyrone

[0083] 5.1 Biochemical experiments

[0084] Glycation reaction refers to the excess sugar being free in the blood, attached to biomacromolecules such as proteins, undergoing condensation, rearrangement, and chain extension reactions to generate highly reactive glycation intermediates; continued action to generate irreversible glycation end-products (AGEs). In the biochemical experiment, glucose solution was mixed with bovine serum albumin (BSA) solution, aminoguanidine was used as a positive control, and phosphate buffer was used as a blank control. The mixed samples were reacted at 55°C in the dark for 4 days, and the fluorescence intensity of the solution product was measured by a fluorescence microplate reader under the conditions of an excitation wavelength of 370nm and an emission wavelength of 440nm, and the inhibition rate was calculated. The anti-glycation effect of the sample was judged by the inhibition rate.

[0085] 5.1.1 Experimental methods:

[0086] (1) Preparation of 2× saccharification reaction solution: Weigh 8 g of bovine serum albumin (Sigma, M26HS122527) and 24 g of glucose (Sigma, SLBR0902V), and use 100 mL of 0.1 mol / L PBS buffer solution (Boster, WH0621P201) to prepare a mixed solution containing 80 mg / mL bovine serum albumin and 240 mg / mL glucose. Filter through a 0.22 μm filter membrane and set aside.

[0087] (2) Preparation of 2× sample solutions: Example 1 was prepared using PBS buffer to prepare 0.0356 mg / mL, 0.0178 mg / mL, 0.0089 mg / mL, 0.00445 mg / mL, and 0.002225 mg / mL solutions for later use.

[0088] (3)Perform the operations as shown in the table below.

[0089] Table 1

[0090] Unit(mL) A B C D 2× saccharification reaction solution 1 0 1 0 2× sample / reference solution 1 1 0 0 PBS / sample solvent 0 1 1 2

[0091] After mixing evenly with a vortex oscillator (Shanghai Qiqian Electronic Technology, MZX-2500), incubate in a 55°C constant temperature incubator (Tianjin Test Instrument Co., Ltd., DH400BII) for 4 days. PBS or sample solvent was used instead of the sample as a negative control or solvent control, 10 mg / mL aminoguanidine hydrochloride (Aladdin, G1909194) was used as a positive control, and PBS was used instead of the glycosylation reaction solution as a control system. After the reaction was completed, the incubated solution was cooled to room temperature for determination, and 200 μL of the reaction solution was taken in turn and added to a 96-well plate. The AGEs inhibition rate was detected and calculated using a fluorescent microplate reader (BioTek, Epoch) at an excitation wavelength of 370 nm and an emission wavelength of 440 nm. The results are expressed as Mean±SD.

[0092]

[0093] Wherein A is the fluorescence intensity of the saccharification system of the sample group; B is the fluorescence intensity of the PBS solution of the sample group; C is the fluorescence intensity of the saccharification system of the control group; and D is the fluorescence intensity of the PBS solution of the control group.

[0094] 5.1.2. Experimental results:

[0095] Table 2

[0096]

[0097] From the AGEs inhibition experiment, it can be seen that after purpurogenol reacted with the glycosylation reaction solution in the mass concentration range of 1.1125μg / mL to 17.8μg / mL, the AGEs inhibition rate was greater than 90% compared with the control group, and P<0.05. Therefore, it is believed that purpurogenol has a very excellent ability to inhibit the formation of AGEs, and the effect is better than 10mg / mL aminoguanidine hydrochloride.

[0098] 5.2 Immunofluorescence experiments

[0099] Glycation is a non-enzymatic chemical reaction. Proteins react with carbonyl compounds (glucose, fructose, glyoxal, etc.) to form AGEs. Carboxymethyl lysine (CML) is one of the representatives of advanced glycation end products. In the experiment, glyoxal was used to induce glycation of primary human skin fibroblasts, aminoguanidine was used as a positive control, and immunofluorescence staining was used to quantitatively analyze the expression of CML. The anti-glycation effect of the sample was determined by the inhibition rate.

[0100] 5.2.1 Toxicity test:

[0101] 1) Experimental methods

[0102] Cell seeding: skin fibroblasts at 7.5×10 3 The cells were seeded into a 96-well plate at a seeding density of 100 cells / well and incubated overnight in an incubator at 37°C and 5% CO2.

[0103] Experimental groups: The experiment set up a zero-adjustment group, a solvent control group, a positive control group and Example 1.

[0104] In Example 1, five concentration gradients were set, and three replicate wells were set under each concentration gradient.

[0105] Solution preparation: Prepare the solution according to the table below.

[0106] Table 3

[0107]

[0108] Administration: Administration was performed when the cell plating rate in the 96-well plate reached 80% to 90%. 200 μL of DMEM culture medium was added to each well of the solvent control group; 200 μL of 0.04 μg / mL epidermal growth factor (EGF) was added to each well of the positive control group; 200 μL of culture medium containing the corresponding concentration of the example was added to each well of the example group; the zeroing group had no cell inoculation and only 200 μL of cell culture medium was added. After administration, the 96-well plate was placed in an incubator and cultured for 48 hours.

[0109] CCK-8 assay: According to the number of test wells required, add 0.01 mL of CCK-8 working solution to 0.1 mL of DMEM medium without fetal calf serum (FCS) per well. Take out the 96-well plate to be tested, discard the medium, add 0.1 mL of CCK-8 working solution to each well, read the OD value at 450 nm and calculate the cell survival rate.

[0110]

[0111] 2) Experimental results

[0112] Table 4

[0113]

[0114]

[0115] The results of cytotoxicity test show that when the mass concentration of purpurogenol is 0.89 μg / mL, the cell survival rate is 116.09%. Therefore, according to the MTT results, purpurogenol has no cytotoxicity within the concentration range of 0.89 μg / mL.

[0116] 5.2.2 Immunofluorescence experiments

[0117] 1) Experimental methods

[0118] Cell inoculation: skin fibroblasts at 1.5*10 3 The cells were inoculated into a 24-well plate at a density of 10 cells / well and incubated in an incubator at 37°C and 5% CO2 for 48 h.

[0119] Administration: The cells were treated with 0.5mM glyoxal to induce glycation reaction as the negative control group. At the same time, a blank group (NC), a positive control group (glyoxal + 0.1mM aminoguanidine) and a sample group (glyoxal + sample) were set up. The cells were treated for 48 hours. At the end of the experiment, the cells were rinsed once with PBS, fixed with paraformaldehyde (PFA), and then CML fluorescence staining was performed. The cells were blocked with 2.5% bovine serum albumin (BSA) for 1 hour, and then incubated with the primary antibody CML (dilution 1:100) at room temperature for 2 hours, and then the secondary antibody Alexa 568Donkey Anti-Mouse IgG (dilution 1:1000) was incubated at room temperature for 1 h, and finally the sections were sealed with an aqueous mounting medium containing diamidinophenylindole (DAPI).

[0120] Detection: Fluorescence images were taken using a fluorescence microscope (upright fluorescence microscope Leica DM2500, magnification 200 times). All results are expressed as Mean±SD.

[0121] 2) Experimental results

[0122] Table 5

[0123]

[0124] The results showed that glyoxal could induce the expression of advanced glycation end product CML in primary human dermal fibroblasts. After aminoguanidine was used as an anti-glycation positive control to treat primary human dermal fibroblasts, CML expression was significantly inhibited, with an inhibition rate of 55.58%. When the mass concentration of porphyrin was 0.178μg / mL and 0.89μg / mL, it had a certain inhibitory effect on glycation, with inhibition rates of 31.37% and 45.21%, respectively.

[0125] The osidone of the present invention can be used as an intermediate raw material or an effective additive for the preparation of medicines, health foods or skin external preparations, and the skin external preparations are preferably cosmetic compositions, including but not limited to the preparation of products in the form of creams, emulsions, gels, lotions, essences, facial masks, eye creams, aerosols (cleansing bubbles), sprays, shower gels and facial cleansers. The weight percentage in the skin external preparation is 0.000017%-10% (w / w). The preferred weight percentage is 0.000089%-5% (w / w). The more preferred weight percentage is 0.00011%-1% (w / w). The most preferred weight percentage is 0.0017%-0.5% (w / w).

[0126] The following are examples of specific applications of porphyrin in skin external preparations, medicines and health foods, as well as the formulations and preparation methods of these dosage forms. In the following tables, "-" means no addition.

[0127] Application Example 1: Preparation of facial cream

[0128]

[0129]

[0130] Application Example 2: Preparation of Emulsion

[0131]

[0132] Application Example 3: Preparation of Gel

[0133]

[0134] Application Example 4: Preparation of lotion

[0135]

[0136]

[0137] Application Example 5: Preparation of Essence

[0138]

[0139] Application Example 6: Preparation of facial mask

[0140]

[0141]

[0142] Application Example 7: Preparation of Eye Cream

[0143]

[0144]

[0145] Application Example 8: Preparation of Aerosol (Cleaning Bubble)

[0146]

[0147] Application Example 9: Preparation of Spray

[0148]

[0149] Application Example 10: Preparation of Shower Gel

[0150]

[0151] Application Example 11: Preparation of facial cleanser

[0152]

[0153]

Claims

1. Anti-glycation application of porphyrin ketone.

2. The use according to claim 1, characterized in that The concentration of porphyrone used is at least 0.1 μg / mL.

3. The use according to claim 2, characterized in that The concentration of porphyrin is at least 0.5 μg / mL.

4. The use according to claim 1, characterized in that The anti-glycation is the end product of inhibiting non-enzymatic glycosylation reaction.

5. The use according to claim 1, characterized in that The anti-glycation is to inhibit the glycation end products.

6. Application of porphyrin in the preparation of external skin preparations, medicines and health foods with anti-glycation effects.

7. The use according to claim 6, characterized in that The content of porphyrin in external skin preparations, medicines and health foods is 0.00001-20% by weight.

8. The use according to claim 7, characterized in that The content of porphyrin in external skin preparations, medicines and health foods is 0.0001-5% 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 claim 6, characterized in that The anti-glycation is the end product of inhibiting non-enzymatic glycosylation reaction.

11. The use according to claim 6, characterized in that The anti-glycation is to inhibit the glycation end products.

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