CN111228139A

CN111228139A - Whitening and moisturizing skin care product

Info

Publication number: CN111228139A
Application number: CN202010171890.8A
Authority: CN
Inventors: 汤涛; 邓艳; 宋艺; 东硕
Original assignee: Shenzhen Baiji Biotechnology Co Ltd
Current assignee: Shenzhen Baiji Biotechnology Co Ltd
Priority date: 2020-03-12
Filing date: 2020-03-12
Publication date: 2020-06-05
Anticipated expiration: 2040-03-12
Also published as: CN111228139B

Abstract

The invention discloses a whitening and moisturizing skin care product, which comprises a solvent, transdermal peptide and small interfering RNA aiming at a P 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 RNA aiming at the P gene is 1g of solvent: 0.1-0.5mg transdermal peptide: 10-50ng of a small interfering nucleic acid directed against the P gene. The whitening and moisturizing skin care product can also comprise 2-5% of arbutin.

Description

Whitening and moisturizing skin care product

Technical Field

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

Background

Andrew z.fire and Craig c.mello, in 1998, have together discovered mechanisms of action for RNA interference in vivo and have collectively won the nobel prize for physiology in 2006. Thus opening a door for the development of a new generation of drugs (RNA interference drugs) for resisting serious diseases such as viruses, cancers and the like. The RNA interference medicine has the advantages of novel action mode, definite action mechanism, strong targeting property, small side effect and the like. RNA interference (RNAi) is the phenomenon whereby double-stranded RNA molecules turn off or silence the expression of homologous genes at the mRNA level. RNA interference techniques, also referred to figuratively as gene knock-down or gene silencing (geneticing), are a typical method of post-transcriptional gene regulation, also known as post-transcriptional gene silencing (PTGS). The first reports on RNA interference appeared in 1990, where RNA interference in transgenic plants was reported simultaneously by two different groups, and later in almost all eukaryotes such as nematodes, drosophila, zebrafish and mice. In 1999, Hamilton and Baulcombe detected RNA fragments of 21-25 nucleotides in length in plants undergoing RNA interference, which were shown to be essential for RNA interference, and were called small interfering nucleic acids (siRNA). The double-stranded siRNA forms an RNA-induced silencing complex (RISC) with associated enzymes and proteins of cell origin. During RNA interference, the sense strand of the double-stranded siRNA is excluded from the complex, the antisense strand directs RISC to bind to the homologous site of the target mRNA, and the target mRNA is then degraded by ribonuclease III in the complex, thereby turning off the expression of the target gene.

There are three main factors determining skin color: 1. melanin, carotene, and other pigment content in the skin; 2. skin thickness and light scattering phenomena on the skin surface; 3. the oxygen and hemoglobin content of blood and reduced hemoglobin in blood vessels in the dermal layer of the skin. Among them, melanin plays a major role, and thus the approach of whitening is mainly based on melanin.

The physiological and biochemical processes of melanin formation can be summarized as: melanin is produced by melanocytes located in the basal layer of the epidermis, which account for about 10% of basal cells, each of which extends via dendrites to adjacent basal and spinous cells, transporting melanin granules. Melanin granules transferred to keratinocytes travel with the epidermal cells up to the stratum corneum, thereby affecting the color of the skin and forming color spots, and are finally excreted as the stratum corneum falls off. Melanin production begins with tyrosine. Tyrosine catalyzes the production of melanin in melanocytes by tyrosinase and various oxidases. The whitening effect can be achieved only by blocking melanin generation or transfer when whitening is required, so most of various whitening components achieve the whitening effect by inhibiting, blocking or influencing melanin.

The enzyme is a biocatalyst, and can inhibit melanin production to effectively reduce melanin amount by inhibiting enzyme essential for each link of melanin production process in cells. At present, most of whitening components take tyrosinase in the first two steps of melanin synthesis reaction as a target spot, and the activity of the tyrosinase is inhibited to achieve the whitening effect. Arbutin is a commonly used tyrosinase inhibitor. Arbutin is mainly extracted from the leaves of Bearberry (Bearberry), and the presence of arbutin can also be found in some fruits and other plants. It can quickly permeate into skin, effectively inhibit the activity of tyrosinase in the skin and block the formation of melanin while not influencing the cell proliferation concentration, and can accelerate the decomposition and excretion of melanin by directly combining with tyrosinase by itself, thereby reducing the skin pigmentation, removing color spots and spots, and has no toxic, irritant, allergic and other side effects on melanocyte, and also has the functions of sterilization and anti-inflammation.

The P gene, also called OCA2 gene, is responsible for the synthesis of the P protein. The P protein is a transmembrane protein of IlOkDa, is composed of 838 amino acid residues, is located on the melanosome membrane, has a contrivance to the integrity of the melanosome membrane, and is a protein necessary for producing melanosome. In addition, research results show that the protein coded by the P gene has obvious homology with membrane transport proteins transported by anions of some of the ginseng chickens, the protein serving as a membrane channel protein can reduce the concentration of protons in melanosomes, and the pH value of the ginseng chickens and the melanosomes is adjusted, so that the local environment is neutral, and the protein is favorable for maintaining the stability of a high-molecular tyrosine-TYRP 1-TYRP2 complex, enhancing the activity of tyrosinase and increasing the generation and deposition of melanin. Thus, inhibition of the P gene can reduce melanin production.

The skin is the largest and most invasive organ, and transdermal administration has the advantages of easy operation, small injury, and the like, and prevents the digestion and degradation of gastrointestinal tracts and livers. Inhibition of the expression of the P gene (OCA2 gene) through the skin may be an effective approach for skin whitening.

At present, the market has a plurality of products for beautifying and protecting skin, and mainly comprises 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 solution for realizing skin whitening by using a skin care product containing a small interfering nucleic acid molecule aiming at a P gene (OCA2 gene).

The skin whitening and moisturizing skin care product provided by the invention comprises a solvent, transdermal peptide (also called transdermal decapeptide-4 in the invention) and small interfering nucleic acid (P gene siRNA) aiming at a P 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, transdermal decapeptide-4 and P gene siRNA is 1g of solvent: 0.1-0.5mg transdermal decapeptide-4: 10-50ng P gene siRNA. 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 product provided by the invention can further comprise arbutin with the concentration of 2-5%.

The invention also provides a preparation method of the whitening and moisturizing skin care product, which comprises the following steps of adding raw materials for preparing a solvent into a water phase pot of a homogenizing and emulsifying machine according to the following formula, and stirring at 400 revolutions per minute: 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, and adding transdermal decapeptide-4 and P gene siRNA after stirring for twenty minutes, wherein the ratio of the three is 1g of solvent: 0.1-0.5mg transdermal decapeptide-4: 10-50ng P gene siRNA, and stirring for ten 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 polysaccharides, 0.003 g of dipotassium glycyrrhizinate, 0.005 g of sodium hyaluronate and 0.002 g of phenoxyethanol, adding transdermal decapeptide-4, P gene siRNA and arbutin after stirring for twenty minutes, wherein the ratio of the four is 1g of solvent: 0.1-0.5mg transdermal decapeptide-4: 10-50ng P gene siRNA: 2-5% of arbutin, and stirring for ten 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:

the hyaluronic acid has very strong water absorption capacity, and the affinity adsorbed water is about 500-1000 times of the mass of the hyaluronic acid, and is a well-known natural moisturizing factor.

The sequence of transdermal decapeptide-4 was synthesized from Guangzhou Yuanji Biotechnology, Inc., as shown in Chinese granted patent CN 106084006B.

Arbutin used in the present invention, CAS No.: 497-76-7, available from Guangzhou Yuanji Biotechnology Ltd

P gene siRNA was synthesized by lebo biotechnology limited, guangzhou city.

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 P 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 siRNA aiming at the P gene is designed, and can pertinently reduce or even silence the expression of the P gene, so that the content of melanin in skin is reduced, and the skin whitening effect is achieved.

The embodiment of the invention discloses a method for designing and obtaining small interfering nucleic acid molecules aiming at a P gene and a method for preparing a whitening and moisturizing skin care product by applying the small interfering molecules. Those skilled in the art can now appreciate from this disclosure that appropriate modifications of the process parameters and adjuvants/starting components can be made, and it is specifically intended that all such similar substitutes and modifications which are obvious to those skilled in the art are deemed to be within the scope of the invention. While the small interfering nucleic acid and skin care compositions of the present invention have been described in terms of preferred embodiments, it will be apparent to those of ordinary skill in the art that variations and modifications of the products and methods described herein, or appropriate variations and combinations thereof, may be made to implement and use the techniques of the present invention without departing from the spirit and scope of the invention. The following examples are merely illustrative, and the present invention is not limited to these examples.

The invention designs and screens active small interfering RNA (siRNA) sequences based on P Gene (NC-000015.10, Gene ID:4948) sequences shown in SEQ ID Nos. 1-4.

P-siRNA 1: sense strand 5 '-GAUCUUCCAAAACAUUUGGAdT-3' SEQ ID No.1

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

P-siRNA 2: sense strand 5 '-GAGCGAAGAAUACCUGCAUAADTdT-3' SEQ ID No.3

Antisense chain 5 '-UUAGCAGUAUCUUCCGCUCdTdT-3' SEQ ID No.4

P-siRNA 3: sense strand 5 '-GCGGAGGUGCGGACCUUAADTdT-3' SEQ ID No.5

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

P-siRNA4: sense strand 5 '-CCUGGAGAAAGAUCUGCAAdTdT-3' SEQ ID No.7

Antisense chain 5 '-UUGCAGAAUCUUUCCAGGGdTdT-3' SEQ ID No.8

The specific steps of the chemical synthesis of the small interfering RNA of the P gene nucleotide sequence are as follows:

(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 '-0-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' -0-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 (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.OM 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.

Example 1

Adding the raw materials for preparing the solvent into a water phase pot of a homogenizing and emulsifying machine according to the following formula, and stirring at 400 revolutions per minute: 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, and after stirring for 20 minutes, transdermal decapeptide-4 and P-siRNA1 are added, wherein the proportion of the three is 1g of solvent: 0.1mg transdermal decapeptide-4: 10ng of P-siRNA1, and stirring for 10 minutes to obtain the whitening and moisturizing skin care product of example 1.

Example 2

Essentially the same as example 1, except that: the proportion of the three is 1g of solvent: 0.1mg transdermal decapeptide-4: 25ng of P-siRNA1, the whitening and moisturizing skin care product of example 2 was obtained.

Example 3

Essentially the same as example 1, except that: the proportion of the three is 1g of solvent: 0.1mg transdermal decapeptide-4: 50ng of P-siRNA1, the whitening and moisturizing skin care product of example 3 was obtained.

Example 4

Essentially the same as example 1, except that: the proportion of the three is 1g of solvent: 0.5mg transdermal decapeptide-4: 10ng of P-siRNA1, the whitening and moisturizing skin care product of example 4 was obtained.

Example 5

Essentially the same as example 1, except that: the proportion of the three is 1g of solvent: 0.5mg transdermal decapeptide-4: 25ng of P-siRNA1, the whitening and moisturizing skin care product of example 5 was obtained.

Example 6

Essentially the same as example 1, except that: the proportion of the three is 1g of solvent: 0.5mg transdermal decapeptide-4: 50ng of P-siRNA1, the whitening and moisturizing skin care product of example 6 was obtained.

Example 7

Essentially the same as example 1, except that: the P-siRNA1 is replaced by arbutin, and the proportion of the P-siRNA1 to arbutin is 1g of solvent: 0.1mg transdermal decapeptide-4: 2% of arbutin to obtain the whitening and moisturizing skin care product of example 7.

Example 8

Essentially the same as example 1, except that: the P-siRNA1 is replaced by arbutin, and the proportion of the P-siRNA1 to arbutin is 1g of solvent: 0.1mg transdermal decapeptide-4: 5% of arbutin to obtain the whitening and moisturizing skin care product of example 8.

Example 9

Essentially the same as example 1, except that: the P-siRNA1 is replaced by arbutin, and the proportion of the P-siRNA1 to arbutin is 1g of solvent: 0.5mg transdermal decapeptide-4: and 2% of arbutin to obtain the whitening and moisturizing skin care product of example 9.

Example 10

Essentially the same as example 1, except that: the P-siRNA1 is replaced by arbutin, and the proportion of the P-siRNA1 to arbutin is 1g of solvent: 0.5mg transdermal decapeptide-4: 5% of arbutin, the whitening and moisturizing skin care product of example 10 was obtained.

Example 11

Adding the raw materials for preparing the solvent into a water phase pot of a homogenizing and emulsifying machine according to the following formula, and stirring at 400 revolutions per minute: 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, transdermal decapeptide-4, P-siRNA1 and arbutin are added after stirring for 20 minutes, the ratio of the four is 1g of solvent: 0.1mg transdermal decapeptide-4: 10ng P-siRNA 1: 2% of arbutin to obtain the whitening and moisturizing skin care product of example 11.

Example 12

Essentially the same as example 11, except that: the ratio of the four is 1g of solvent: 0.1mg transdermal decapeptide-4: 10ng P-siRNA 1: 5% of arbutin, the whitening and moisturizing skin care product of example 12 is obtained.

Example 13

Essentially the same as example 11, except that: the ratio of the four is 1g of solvent: 0.1mg transdermal decapeptide-4: 25ng P-siRNA 1: and 2% of arbutin to obtain the whitening and moisturizing skin care product of example 13.

Example 14

Essentially the same as example 11, except that: the ratio of the four is 1g of solvent: 0.1mg transdermal decapeptide-4: 25ng P-siRNA 1: 5% of arbutin, the whitening and moisturizing skin care product of example 14 is obtained.

Example 15

Essentially the same as example 11, except that: the ratio of the four is 1g of solvent: 0.1mg transdermal decapeptide-4: 50ng P-siRNA 1: and 2% of arbutin to obtain the whitening and moisturizing skin care product of example 15.

Example 16

Essentially the same as example 11, except that: the ratio of the four is 1g of solvent: 0.1mg transdermal decapeptide-4: 50ng P-siRNA 1: 5% of arbutin, the whitening and moisturizing skin care product of example 16 was obtained.

Example 17

Essentially the same as example 11, except that: the ratio of the four is 1g of solvent: 0.5mg transdermal decapeptide-4: 10ng P-siRNA 1: and 2% of arbutin to obtain the whitening and moisturizing skin care product of example 17.

Example 18

Essentially the same as example 11, except that: the ratio of the four is 1g of solvent: 0.5mg transdermal decapeptide-4: 10ng P-siRNA 1: 5% of arbutin, the whitening and moisturizing skin care product of example 18 was obtained.

Example 19

Essentially the same as example 11, except that: the ratio of the four is 1g of solvent: 0.5mg transdermal decapeptide-4: 25ng P-siRNA 1: and 2% of arbutin to obtain the whitening and moisturizing skin care product of example 19.

Example 20

Essentially the same as example 11, except that: the ratio of the four is 1g of solvent: 0.5mg transdermal decapeptide-4: 25ng P-siRNA 1: 5% of arbutin, the whitening and moisturizing skin care product of example 20 was obtained.

Example 21

Essentially the same as example 11, except that: the ratio of the four is 1g of solvent: 0.5mg transdermal decapeptide-4: 50ng P-siRNA 1: and 2% of arbutin to obtain the whitening and moisturizing skin care product of example 21.

Example 22

Essentially the same as example 11, except that: the ratio of the four is 1g of solvent: 0.5mg transdermal decapeptide-4: 50ng P-siRNA 1: 5% of arbutin, the whitening and moisturizing skin care product of example 22 is obtained.

Example 23

Substantially the same as example 22 except that P-siRNA1 was replaced by P-siRNA2, the skin whitening and moisturizing cosmetic of example 23 was obtained.

Example 24

Substantially the same as in example 22 except that P-siRNA1 was replaced with P-siRNA3, the whitening and moisturizing skin care product of example 24 was obtained.

Example 25

Substantially the same as in example 22 except that P-siRNA1 was replaced with P-siRNA4, the whitening and moisturizing skin care product of example 25 was obtained.

Example 26 experiment for melanin inhibition of mouse skin

The whitening and moisturizing skin care products 1-25 were obtained in the examples shown above, and the control group containing only the vehicle was 26. 130 adult female C57BL/6 mice were housed in an environmentally controlled animal care laboratory using a standard mouse diet. The hair was carefully trimmed with scissors in a 1cm x 1cm area of the abdomen (without using any mice with evidence of skin damage). Mice were selected and randomized into different groups of 5 mice each. Each group of skin care products was applied once a day to the area of the abdomen where hair was removed. After 7 days of application of the above formulation, mice were sacrificed and the skin was cut into thin slices to measure melanin inhibiting activity. To extract melanin, the cell suspension of the skin tissue mass was shaken for 60 minutes at 80 ℃ with an equal volume of a mixture containing 10% dimethyl sulfoxide and 1N NaOH. After centrifugation at 2,000rpm for 10 minutes, the OD was measured at 475nm using a SpectroQuest Reader, and a higher OD indicates more melanin production.

TABLE 1 Effect of inhibiting melanin for each group

According to the data shown in the table above, the OD value is highest in the case of only skin care product solvent, indicating that the skin contains the most melanin; and after 7 days of using the skin care product provided by the invention, the melanin in the skin of the mouse is obviously reduced, and the combination of 22, namely 1g of solvent, 0.5mg of transdermal peptide and 50ng of P gene siRNA No.1 has the best effect of inhibiting the melanin by 5 percent arbutin.

Example 27 moisturizing Effect test on skin

Skin stratum corneum hydration test: the working principle is that the capacitor is used as an instrument probe, and as water is a substance with the largest dielectric constant on the skin, when the moisture content of the skin changes, the capacitance value of the skin also changes, so that the moisture content on the surface of the skin can be analyzed by measuring the capacitance value of the skin. A commonly used instrument is Corneometer CM 825.

The experimental method comprises the following steps: marking out 3X 3cm on the bent side of the left and right forearms of the person to be tested²In a square experiment area with the size, 1ml of the whitening and moisturizing skin care product prepared in the embodiment 1-25 of the invention (a control group only containing solvent is 26) is uniformly coated on the left arm serving as a test area of the moisturizing product, a corresponding symmetric area of the right arm serves as a blank control, then the moisture content of each test part is tested by a Corneometer CM825 and repeated for 5 times respectively to obtain an average value; the group containing only vehicle was 26. The hydration rate was calculated by the following formula.

Hydration ═ (test value-blank value)/blank value × 100%

Table 2 moisturizing effect test results

As shown in the above table, the skin care products provided by the present invention have a longer moisturizing time and a better moisturizing effect than the vehicle group, and particularly the moisturizing effects of examples 21 and 22 are the best moisturizing effects.

In conclusion, the skin care product provided by the invention has whitening and moisturizing effects. The combined melanin inhibiting and moisturizing effects are best achieved in example 22, i.e., 1g of vehicle, 0.5mg of transdermal peptide, 50ng of P gene siRNA No.1, and 5% arbutin.

Sequence listing

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Claims

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

2. The whitening and moisturizing skin care product according to claim 1, further comprising arbutin.

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

P-siRNA 1: sense strand 5 '-GAUCUUCCAAAACAUUUGGAdTdT-3'

Antisense strand 5 '-UCCAAUGUUUGAGAUCdTdT-3'

Or P-siRNA 2: sense strand 5 '-GAGCGAAGAAUACCUGCAUAADTdT-3'

Antisense chain 5 '-UUAGCAGUAUCUUCCGCUCdTdT-3'

Or P-siRNA 3: sense strand 5 '-GCGGAGGUGCGGACCUUAADTdT-3'

Antisense strand 5 '-UUAAAGGUCCGCACCUCCGCTdTdT-3'

Or P-siRNA4 sense strand 5 '-CCUGGAAAGAUCUGCAAdTdT-3'

The antisense chain is 5 '-UUGCAGAAUCUUUCCAGGGdTdT-3'.

4. The whitening and moisturizing skin care product of claim 1, wherein 1g of the skin care product solvent contains 0.1-0.5mg of transdermal peptide and 10-50ng of small interfering nucleic acid aiming at the P gene.

5. The whitening and moisturizing skin care product of claim 2, wherein 1g of the skin care product solvent contains 0.1-0.5mg of transdermal peptide, 10-50ng of small interfering nucleic acid aiming at the P gene and 2-5% of arbutin.

6. The whitening and moisturizing skin care product of claim 1, wherein 1 gram of the skin care product 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 butanediol, 0.001 gram of taurine, 0.001 gram of potassium methoxysalicylate, 0.001 gram of glyceryl caprylate, 0.002 gram of caprylyl hydroximic 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 product of 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 product.

8. A preparation method of a whitening and moisturizing skin care product 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 r/m for twenty minutes, and then the transdermal peptide and small interfering nucleic acid aiming at a P gene are added, wherein the proportion of the three is 1g of solvent: 0.1-0.5mg transdermal peptide: 10-50ng of small interfering nucleic acid against the P gene, and stirring was continued for another ten minutes.

9. A preparation method of a whitening and moisturizing skin care product 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 r/m for twenty minutes, then the transdermal peptide, the small interfering nucleic acid aiming at the P gene and the arbutin are added, and the proportion of the four solvents is 1 g: 0.1-0.5mg transdermal peptide: 10-50ng of a small interfering nucleic acid directed against the P gene: 2-5% of arbutin, and stirring for ten minutes.