阅读说明：本技术 一种美白保湿护肤品组合物 (Whitening and moisturizing skin care product composition ) 是由 汤涛 邓艳 宋艺 东硕 于 2020-07-02 设计创作，主要内容包括：本发明公开了一种美白保湿护肤品,包括溶媒、透皮肽和针对MATP基因的小干扰核酸,其中溶媒由下述原料制备而成：水、甘油、凝血酸、谷胱甘肽、丁二醇、牛磺酸、甲氧基水杨酸钾、甘油辛酸酯、辛酰羟肟酸、对羟基苯乙酮、肌肽、产碱杆菌多糖类、甘草酸二钾、透明质酸钠、苯氧乙醇；溶媒、透皮肽和针对MATP基因的小干扰核酸的比例为1g溶媒∶0.1-0.5mg透皮肽∶10-50ng针对MATP基因的小干扰核酸。本发明的美白保湿护肤品还可以包括2-5％的烟酰胺。(The invention discloses a whitening and moisturizing skin care product, which comprises a solvent, transdermal peptide and small interfering nucleic acid aiming at MATP gene, wherein the solvent is prepared from the following raw materials: water, glycerol, tranexamic acid, glutathione, butanediol, taurine, potassium methoxysalicylate, glyceryl caprylate, caprylyl hydroximic acid, p-hydroxyacetophenone, carnosine, Alcaligenes polysaccharide, dipotassium glycyrrhizinate, sodium hyaluronate and phenoxyethanol; the ratio of the solvent, the transdermal peptide and the small interfering nucleic acid aiming at the MATP gene is 1g of solvent, 0.1-0.5mg of transdermal peptide and 10-50ng of small interfering nucleic acid aiming at the MATP gene. The whitening and moisturizing skin care product can also comprise 2-5% of nicotinamide.)

1. A skin-whitening and moisturizing skin care product composition is characterized by comprising a skin care product solvent, transdermal peptide and small interfering ribonucleic acid aiming at MATP gene.

2. The whitening and moisturizing skin care composition according to claim 1, further comprising niacinamide.

3. The whitening and moisturizing skin care product according to claim 1, wherein the small interfering nucleic acid directed against the MATP gene is selected from the group consisting of:

MATP-siRNA 1: sense strand 5 '-GCCUACUUAUUGUGUCUdTdT-3'

Antisense strand 5 '-AGACAUUCAAAUAAGUAGGCdTdT-3'

Or MATP-siRNA 2: sense strand 5 '-GGUUCCUAUCGAGAAAGUUAdTdT-3'

Antisense chain 5 '-UAACUUCUCGAUAGAACCdTdT-3'

Or MATP-siRNA 3: sense strand 5 '-CGUCAAAUCCAGUUGAAAdTdT-3'

Antisense strand 5 '-UUCAAACUGGAUUGACGdTdT-3'

Or MATP-siRNA 4: sense strand 5 '-GCUUGAUUGCUAACCCAATdT-3'

The antisense strand is 5 '-UUGGGGUUAGCAAUCAAAGCdTdT-3'.

4. The whitening and moisturizing skin care composition as claimed in claim 1, wherein 1g of the skin care vehicle contains 0.1-0.5mg of transdermal peptide and 10-50ng of small interfering nucleic acid against MATP gene.

5. The whitening and moisturizing skin care composition as claimed in claim 2, wherein 1g of the skin care vehicle contains 0.1-0.5mg of transdermal peptide, 10-50ng of small interfering nucleic acid against MATP gene and 2-5% of niacinamide.

6. The whitening and moisturizing skin care composition according to claim 1, wherein 1 gram of the skin care vehicle comprises 0.925 gram of water, 0.05 gram of glycerin, 0.003 gram of tranexamic acid, 0.002 gram of glutathione, 0.001 gram of butylene glycol, 0.001 gram of taurine, 0.001 gram of potassium methoxysalicylate, 0.001 gram of glyceryl caprylate, 0.002 gram of caprylyl hydroxamic acid, 0.001 gram of p-hydroxyacetophenone, 0.001 gram of carnosine, 0.002 gram of alcaligenes polysaccharides, 0.003 gram of dipotassium glycyrrhizinate, 0.005 gram of sodium hyaluronate, and 0.002 gram of phenoxyethanol.

7. The whitening and moisturizing skin care composition according to any one of claims 1-2, wherein the melanin content in the skin is reduced by 20% or more after 7 days of use of the skin care composition.

8. A preparation method of a whitening and moisturizing skin care product composition is characterized in that 92.5 parts by weight of water, 5 parts of glycerol, 0.3 part of tranexamic acid, 0.2 part of glutathione, 0.1 part of butanediol, 0.1 part of taurine, 0.1 part of potassium methoxysalicylate, 0.1 part of glyceryl caprylate, 0.2 part of caprylyl hydroximic acid, 0.1 part of p-hydroxyacetophenone, 0.1 part of carnosine, 0.2 part of alcaligenes polysaccharides, 0.3 part of dipotassium glycyrrhizinate, 0.5 part of sodium hyaluronate and 0.2 part of phenoxyethanol are added into a water phase pot of a homogenizing and emulsifying machine to be stirred at 400 revolutions per minute, and after being stirred for twenty minutes, the transdermal peptide and small interfering nucleic acid aiming at MATP gene are added, wherein the proportion of the three solvents is 1 g: 0.1-0.5mg transdermal peptide: 10-50ng of small interfering nucleic acid against MATP gene, and stirring was continued for another ten minutes.

9. A preparation method of a whitening and moisturizing skin care product composition is characterized in that 92.5 parts by weight of water, 5 parts of glycerol, 0.3 part of tranexamic acid, 0.2 part of glutathione, 0.1 part of butanediol, 0.1 part of taurine, 0.1 part of potassium methoxysalicylate, 0.1 part of glyceryl caprylate, 0.2 part of caprylyl hydroximic acid, 0.1 part of p-hydroxyacetophenone, 0.1 part of carnosine, 0.2 part of alcaligenes polysaccharides, 0.3 part of dipotassium glycyrrhizinate, 0.5 part of sodium hyaluronate and 0.2 part of phenoxyethanol are added into a water phase pot of a homogenizing and emulsifying machine to be stirred at 400 revolutions per minute, and after being stirred for twenty minutes, the transdermal peptide, the small interfering nucleic acid aiming at MATP gene and the nicotinamide are added, wherein the proportion of the four solvents is 1 g: 0.1-0.5mg transdermal peptide: 10-50ng of small interfering nucleic acid against MATP gene: 2-5% nicotinamide, and stirring for another ten minutes.

Technical Field

The invention relates to the technical field of cosmetics, and particularly relates to a whitening and moisturizing skin care product composition and a preparation method thereof.

Background

Melanin (melanin) is produced by melanocytes of the skin, hair follicles, iris, and retina. When stimulated by factors such as ultraviolet rays and the like, the melanocyte can synthesize a large amount of tyrosinase which is necessary for melanin biosynthesis, tyrosine is converted into dopaquinone under the catalysis of the tyrosinase, the dopaquinone is further oxidized to form dopachrome, and finally the melanin is formed. Excessive melanin causes skin to become black, and non-uniformly distributed melanin causes chloasma and freckle to be generated. The earliest method of skin whitening was a peeling bleaching method. Different peeling bleaching formulas can carry out peeling in different degrees, and the common formula is some acids and alcohols, has certain dangerousness and is not suitable for being used as a manufacturing method of common cosmetics. It is then gradually recognized that the difference in dark and light skin color is related to the level of tyrosinase activity in melanocytes, and thus inhibitor products containing various tyrosinase enzymes are continuously emerging. Common tyrosinase inhibitors are hydroquinone, kojic acid, arbutin, ascorbic acid and more recently biological products. In addition, new ideas began to emerge.

Niacinamide is a derivative of vitamin B3. Whitening mainly interferes with the synthesis of melanin, can prevent the generated melanin from being transported to an epidermal layer, can accelerate the metabolism of the epidermal layer, and can lead cells containing the melanin to be dropped. In general, nicotinamide is said to have no way for melanin particles that have already been synthesized to be transported out. The skin is not blackened no matter how much melanin is produced by the melanocyte but can not be transported out, thereby achieving the whitening effect. On the one hand, niacinamide has the functions of moisturizing and repairing skin barriers. One important component of the skin barrier is ceramide, and topical niacinamide can increase the levels of ceramide and free fatty acids, prevent water loss from the skin, promote microcirculation in the dermis, and enhance the skin barrier.

Gene whitening is a new concept that is different from existing methods and active ingredients used for whitening. It controls the expression of genes involved in melanin formation from the gene level. The expression level of the gene related to melanin formation is obviously reduced but is different from the related gene defect or deletion in albinism. RNA interference (RNAi) is a phenomenon in which the expression of a specific gene is inhibited in an organism, and it refers to a phenomenon in which, when double-stranded RNA homologous to the coding region of an endogenous mRNA is present in a cell, the mRNA is degraded, resulting in silencing of the gene expression. Small RNA molecules 21-23nt in length, called small interfering RNA (siRNA), are directly responsible for RNA interference phenomena, and siRNA is considered to be a rapid and efficient method for regulating gene expression in vivo. The MATP gene is closely related to pigmentation, and the MATP protein coded by the MATP gene plays a decisive role in mammalian pigmentation, so that the expression of the MATP gene is specifically inhibited through RNA interference, the activity of the MATP gene is silenced, and the MATP gene is an effective way for inhibiting melanin synthesis and improving skin color.

At present, a plurality of products for beautifying and protecting skin are available on the market, and mainly comprise products for resisting aging, beautifying, whitening and the like. These products have an improving effect on skin aging, cortical aging, darkening of color, dullness, etc., which are caused by environmental changes and industrial pollution. However, most of the products are chemical preparations, the water solubility is poor, so that the whitening and anti-aging effects can be really and effectively achieved by using a large amount of the chemical preparations or for a long time, and the chemical preparations have the disadvantages of great side effects and high price and are difficult to achieve the aim of treating both symptoms and root causes.

Disclosure of Invention

In order to solve the problems, the invention provides a skin care product composition aiming at the small interfering nucleic acid molecule of the MATP gene to realize the solution of skin whitening and moisturizing.

The skin-whitening and moisturizing skin care product composition provided by the invention comprises a skin care product solvent, transdermal peptide (also called transdermal decapeptide-4) and small interfering nucleic acid (MATP gene siRNA) aiming at MATP gene, wherein the skin care product solvent comprises the following raw materials: water, glycerol, tranexamic acid, glutathione, butanediol, taurine, potassium methoxysalicylate, glyceryl caprylate, caprylyl hydroximic acid, p-hydroxyacetophenone, carnosine, Alcaligenes polysaccharide, dipotassium glycyrrhizinate, sodium hyaluronate and phenoxyethanol; the ratio of the solvent, the transdermal peptide and the small interfering nucleic acid aiming at the MATP gene is 1g of solvent, 0.1-0.5mg of transdermal peptide and 10-50ng of small interfering nucleic acid aiming at the MATP gene. Specifically, 1g of the vehicle comprises the following raw materials:

0.925 g of water, 0.05 g of glycerol, 0.003 g of tranexamic acid, 0.002 g of glutathione, 0.001 g of butanediol, 0.001 g of taurine, 0.001 g of potassium methoxysalicylate, 0.001 g of glyceryl caprylate, 0.002 g of caprylyl hydroxamic acid, 0.001 g of p-hydroxyacetophenone, 0.001 g of carnosine, 0.002 g of alcaligenes polysaccharides, 0.003 g of dipotassium glycyrrhizinate, 0.005 g of sodium hyaluronate and 0.002 g of phenoxyethanol.

In another embodiment, the whitening and moisturizing skin care composition provided by the invention can further comprise nicotinamide with the concentration of 2-5%.

The invention also provides a preparation method of the whitening and moisturizing skin care product composition, the raw materials for preparing the skin care product solvent are added into a water phase pot of a homogenizing emulsifying machine according to the following weight ratio, stirred at 400 r/m, added with the transdermal peptide and MATP gene siRNA after being stirred for 20 minutes, and the proportion of the transdermal peptide to the MATP gene siRNA is 1g of solvent to 0.1-0.5mg of transdermal peptide to 10-50ng of MATP gene siRNA, and then stirred for 10 minutes.

The invention provides another preparation method of a whitening and moisturizing skin care product, which is characterized in that the raw materials for preparing the solvent are added into a water phase pot of a homogenizing and emulsifying machine according to the following formula and stirred at a speed of 400 r/min: 0.925 g of water, 0.05 g of glycerol, 0.003 g of tranexamic acid, 0.002 g of glutathione, 0.001 g of butanediol, 0.001 g of taurine, 0.001 g of potassium methoxysalicylate, 0.001 g of glyceryl caprylate, 0.002 g of caprylyl hydroxamic acid, 0.001 g of p-hydroxyacetophenone, 0.001 g of carnosine, 0.002 g of alcaligenes polysaccharide, 0.003 g of dipotassium glycyrrhizinate, 0.005 g of sodium hyaluronate and 0.002 g of phenoxyethanol, stirring for 20 minutes, adding transdermal peptide, MATP gene siRNA and nicotinamide, wherein the ratio of the transdermal peptide to the nicotinamide is 1g of solvent to 0.1-0.5mg of transdermal peptide to 10-50ng MATP gene siRNA to 2-5% of nicotinamide, and further stirring for 10 minutes.

The raw materials used for preparing the solvent in the invention are all purchased from Guangzhou Yuanji Biotechnology GmbH, and the catalog numbers of the products are as follows:

water catalog number: 06260

Glycerol catalog number: 02421

Tranexamic acid catalog number: 04866

Glutathione catalog number: 02586

Butanediol catalog number: 0003

Taurine catalog number: 04876

Catalog number of potassium methoxysalicylate: 03354

Catalog number for glyceryl caprylate: 02478

Catalog number for caprylyl hydroxamic acid: 07218

Catalog number of p-hydroxyacetophenone: 02021

Carnosine catalogue number: 03150

Alcaligenes polysaccharide catalogue number: 01616

Catalog number of dipotassium glycyrrhizinate: 02397

Sodium hyaluronate catalog no: 06722

Catalogue number of phenoxyethanol: 01294

The sequence of the transdermal peptide is shown in the Chinese patent No. CN106084006B, and is synthesized by Guangzhou Yuanji Biotech Co.

Nicotinamide used in the present invention was purchased from Guangzhou Yuanji Biotech, Inc., catalog No.: 07359.

MATP gene siRNA was synthesized by Ruibo Biotech, Inc., Guangzhou.

The whitening and moisturizing skin care product has remarkable effect, can enhance the water locking capacity of the skin and reduce the melanin content of the skin, and has the effects of whitening and moisturizing.

The invention has the advantages of

The skin whitening and moisturizing skin care product provided by the invention can efficiently and specifically inhibit the expression of MATP gene in skin, reduce the generation of melanin, enhance the water locking capacity of skin, has low toxic and side effects, can be used for a long time, and achieves the aim of treating both symptoms and root causes.

Detailed Description

The present invention will be further described with reference to the following examples, which are intended to be illustrative only and not to be limiting of the invention in any way, and any person skilled in the art can modify the present invention by applying the teachings disclosed above and applying them to equivalent embodiments with equivalent modifications. Any simple modification or equivalent changes made to the following embodiments according to the technical essence of the present invention, without departing from the technical spirit of the present invention, fall within the scope of the present invention.

The invention designs and screens active small interfering RNA (siRNA) sequences based on MATP Gene (NC-000005.10, Gene ID: 51151), and the MATP Gene siRNA adopted by the invention is shown in SEQ ID No. 1-8. However, it is understood that other active siRNAs against the MATP gene can achieve the technical effects of the present invention and are within the scope of the present invention.

MATP gene small interfering nucleic acid sequence:

MATP-siRNA 1: sense strand 5 '-GCCUACUUAUUGUGUdTdT-3' SEQ ID No.1

Antisense strand 5 '-AGACAUUCAAAUAGUGGCdTdT-3' SEQ ID No.2

MATP-siRNA 2: sense strand 5 '-GGUUCAUCGAGAAGUUADTTdT-3' SEQ ID No.3

Antisense strand 5 '-UAACUUCUCGAUAGAACCdTdT-3' SEQ ID No.4

MATP-siRNA 3: sense strand 5 '-CGUCAAAUCCAGUUGAAAdTdT-3' SEQ ID No.5

Antisense strand 5 '-UUCAAACUGGAUUGACGdTdT-3' SEQ ID No.6

MATP-siRNA 4: sense strand 5 '-GCUUGAUUGCUAACCCAATdT-3' SEQ ID No.7

Antisense strand 5 '-UUGGGGUUAGCAAUCAAAGCdTdT-3' SEQ ID No.8

(1) Synthesis of oligoribonucleotides: synthesis of oligoribonucleotides was performed on an automated DNA/RNA synthesizer. The small interfering RNA consists of a section of 19 oligoribonucleotides and 2 deoxythymidylate. Therefore, the starting material is 5 '-O _ p-dimethoxy-thymidine connected with a solid phase, and the specific synthesis of each cycle can be completed by four steps, wherein the first step is to elute the protecting group at the 5' position on the thymidine connected with the immobilization under the action of trichloroacetic acid; secondly, coupling 5' -O-p-dimethoxytrityl-thymidine phosphoramidite to the previous thymidine which is deprotected under the action of an active catalyst S-ethyltetrazole to form dithymidine phosphite triester, wherein the coupling time and the coupling frequency are all completed according to the program provided by an instrument manufacturer; the third step is that the coupled dithymidine phosphotriester is oxidized into dithymidine phosphotriester under the action of 0.05M iodine water; the fourth step is acetylation, where small amounts of unreacted reactive groups (e.g., hydroxyl and amine groups) on the solid phase are reacted with acetic anhydride to form esters or amides, thereby achieving a blocking effect to reduce the overall byproduct generation, and this cycle is repeated until the synthesis of the entire nucleic acid sequence is completed.

(2) Deprotection: putting the synthesized solid phase small interfering nucleic acid into a bottle which can be sealed, adding 1ml of ethanol/amine (the volume ratio is 1: 3), sealing, putting the bottle in an incubator at 55-70 ℃, incubating for 2-30 hours, taking out the solution, rinsing the solid phase with double distilled water again, collecting the eluent, and drying to remove the solvent. Then, 1ml of tetrahydrofuran solution (IM) of tetrabutylammonium fluoride was added, and the mixture was left at room temperature for 4 to 12 hours, followed by ethanol precipitation to obtain a crude product of small interfering nucleic acid.

(3) And (3) purification and separation: dissolving the crude product of the small interfering nucleic acid in 2 ml of ammonium acetate aqueous solution, then separating by reaction C18 high pressure liquid chromatography, using a gradient elution method to collect the main product of the small interfering nucleic acid (eluent A: 0.IM ammonium acetate; eluent B: 20% of 0.IM ammonium acetate and 80% of acetonitrile), removing the solvent in the main product, adding 5 ml of 80% acetic acid aqueous solution, standing at room temperature for 15 minutes, and then separating the solution by anion exchange (DEAE-5PW, anion exchange column), thus obtaining the small interfering nucleic acid with the purity of more than 90% (gradient elution, eluent A: 0.025M Tris-HCl, 0.025M NaCl, pH 8, 5% acetonitrile; eluent B: 0.025M Tris-HCl, 2.0M NaCl, pH 8, 5% acetonitrile).

(4) Desalting: the purified small interfering RNA is dialyzed to remove salts, and then the small interfering RNA solution is filtered, sterilized, dried and crystallized. Then annealing the oligoribonucleotides of the sense strand and the antisense strand to form a stable double-stranded small interfering RNA, by mixing and dissolving the oligoribonucleotides of the sense strand and the antisense strand in 1-2 ml of buffer (IOmM Tris, pH 7.5-8.0, 50mM NaCl), heating the solution to 95 ℃, slowly cooling the solution to room temperature, and storing the solution in a refrigerator at 4 ℃ for use at any time.