Short oligonucleotide with whitening and / or anti-aging function and application thereof

Abstract

The present application provides a short oligonucleotide with whitening and / or anti-aging function and application thereof, wherein the nucleotide sequence of the short oligonucleotide is shown in any one of SEQ ID NO:1-SEQ ID NO:14. The related sample of the present application can promote SIRT-1 expression at a concentration of 20 μg / mL, can inhibit melanin production at a concentration of 100 μg / mL, and obtain basic data of in-vitro safety evaluation in a concentration range of 0.16-100 μg / mL, and can be used in whitening and / or anti-aging products.

Description

Technical Field

[0001] This invention relates to the field of cosmetics and personal care products technology, specifically to a short oligonucleotide with whitening and / or anti-aging functions and its applications. Background Technology

[0002] Skin whitening, anti-photoaging, and skin repair are important research and development directions in the cosmetics industry. With the upgrading of consumer skincare needs, functional cosmetics that combine whitening and brightening, anti-photoaging, and barrier repair effects have become a hot research topic in the industry.

[0003] In recent years, salmon-derived DNA raw materials have been widely used in skin repair cosmetics due to their good biocompatibility. In existing commercially available products and published patents, these raw materials mostly use a mixture of DNA fragments extracted from salmon testis tissue as the main active ingredient, commonly in the form of PDRN, smPDRN, or DNA-sodium, and are mainly used in skin repair, anti-inflammatory, and cell proliferation promotion applications.

[0004] Existing technology records small-molecule polydeoxyribonucleotides (smPDRNs), with fragment lengths ranging from 50 to 1000 bp, are prepared from salmon testis DNA through enzymatic digestion and fractionation, and are used in skin repair, cell proliferation promotion, and anti-inflammatory applications. This method uses naturally extracted products as raw materials, belonging to a mixture of DNA fragments with a wide length distribution, and does not disclose a chemically synthesized short oligonucleotide system with a clearly defined sequence of 16–22 bp. Patent document CN120859873A discloses a class of synthetic PDRN-related sequences, mainly used to promote VEGF expression and skin repair. This method only focuses on skin repair efficacy, without considering the inhibition of melanin production and the promotion of SIRT-1 expression as core effects, nor does it disclose the same specific short oligonucleotide sequence system and corresponding application pathways as this invention. Patent document CN118546922B discloses the application of fish-derived DNA-sodium in whitening, mainly achieving whitening effects by regulating the tyrosinase pathway. The proposed scheme still uses a mixture of naturally derived DNA, does not specify a particular chemically synthesized short oligonucleotide structure of 16–22 bp, and does not construct a combined application system for whitening, anti-photoaging, and repair that promotes SIRT-1 expression.

[0005] In summary, existing technologies are mostly limited to naturally extracted nucleic acid mixtures or single-function nucleic acid sequence schemes, resulting in several technical shortcomings: First, poor component controllability and insufficient batch stability. Naturally extracted PDRN and DNA-sodium are mixtures of random DNA fragments with unclear base sequences and uneven fragment length distributions, greatly affected by raw material sources and extraction processes, making standardized quality control difficult and leading to significant batch-to-batch variations in product efficacy. Second, limited efficacy dimensions and weak synergistic effects. Most existing technologies focus only on skin repair, anti-inflammation, or single whitening effects, with few solutions that simultaneously inhibit melanin production and regulate SIRT-1 expression using the same nucleic acid active component, failing to meet the combined needs of whitening, anti-photoaging, and multi-effect repair in cosmetics. Third, unclear structure-efficacy correlation. Natural mixed nucleic acid systems cannot establish a correspondence between specific base sequences, fragment lengths, and specific skincare effects, making precise targeted screening of functional fragments difficult, resulting in ambiguous technical definitions and hindering the precise development and application of active components. Fourth, significant challenges in purity and safety control. Natural tissue extraction processes are prone to leaving fish protein residues, endotoxins, and other impurities, increasing the risk of skin sensitization. The purity and safety of raw materials are also difficult to guarantee consistently. Fifth, there is insufficient support regarding the molecular mechanism of action. Current technologies lack clear verification of the SIRT-1 anti-aging target, and the mechanisms of action for anti-photoaging and intrinsic anti-aging are unclear, limiting the efficacy endorsement and application value of products.

[0006] In view of the above-mentioned shortcomings of existing technologies, it is necessary to develop a new nucleic acid activity system to solve the technical defects of existing natural nucleic acid raw materials, such as uncontrollable components, single efficacy, unclear structure-efficacy relationship, poor safety, and lack of clear anti-photoaging mechanism. Summary of the Invention

[0007] This invention provides a chemically synthesized short oligonucleotide derived from the salmon genome and its application in whitening and / or anti-aging cosmetics, aiming to overcome the technical shortcomings of existing natural nucleic acid raw materials, such as uncontrollable components, limited efficacy, unclear structure-efficacy relationship, and poor safety profile. The invention adopts the following technical solution: The present invention proposes a short oligonucleotide, the nucleotide sequence of which is shown in any one of SEQ ID NO:1 to SEQ ID NO:14. The short oligonucleotide can be used to form a double-stranded oligonucleotide or a short oligonucleotide composition having whitening and / or anti-aging related functions.

[0008] The present invention also proposes a double-stranded oligonucleotide with whitening and / or anti-aging functions, comprising at least one of the complementary strand combinations YH01, YH02, YH03, YH04, YH05, YH06, and YH07; The nucleotide sequence of the sense strand in the complementary strand combination YH01 is shown in SEQ ID NO:1, and the nucleotide sequence of the antisense strand is shown in SEQ ID NO:2. The nucleotide sequence of the sense strand of the complementary strand combination YH02 is shown in SEQ ID NO:3, and the nucleotide sequence of the antisense strand is shown in SEQ ID NO:4. The nucleotide sequence of the sense strand of the complementary strand combination YH03 is shown in SEQ ID NO:5, and the nucleotide sequence of the antisense strand is shown in SEQ ID NO:6. The nucleotide sequence of the sense strand of the complementary strand combination YH04 is shown in SEQ ID NO:7, and the nucleotide sequence of the antisense strand is shown in SEQ ID NO:8. The nucleotide sequence of the sense strand of the complementary strand assembly YH05 is shown in SEQ ID NO:9, and the nucleotide sequence of the antisense strand is shown in SEQ ID NO:10. The nucleotide sequence of the sense strand of the complementary strand combination YH06 is shown in SEQ ID NO:11, and the nucleotide sequence of the antisense strand is shown in SEQ ID NO:12. The nucleotide sequence of the sense strand of the complementary strand assembly YH07 is shown in SEQ ID NO:13, and the nucleotide sequence of the antisense strand is shown in SEQ ID NO:14.

[0009] The aforementioned short oligonucleotides can form an equimolar mixture of complementary strands; the equimolar mixture can be annealed to form double-stranded oligonucleotides, or it can be used as a complementary single-stranded mixture without special annealing.

[0010] The annealing process involves placing an equimolar mixture in a common annealing buffer and heating it at 95°C for 5-10 min to fully denature the single strands of nucleic acid; then slowly cooling it to room temperature at a rate of 1-2°C / min to promote the annealing and pairing of the two complementary strands.

[0011] The present invention also proposes a short oligonucleotide composition with whitening and / or anti-aging functions, wherein the components of the short oligonucleotide composition include the above-mentioned short oligonucleotide and / or the above-mentioned double-stranded oligonucleotide with whitening and / or anti-aging functions.

[0012] Furthermore, in the complementary chain combinations YH01, YH02, YH03, YH04, YH05, YH06, and YH07, the sense chain and the antisense chain are mixed in an equimolar ratio.

[0013] The present invention also proposes a product with whitening function, the product comprising the above-mentioned short oligonucleotide, the above-mentioned double-stranded oligonucleotide and / or the above-mentioned short oligonucleotide composition.

[0014] Furthermore, the whitening function includes inhibiting melanin production.

[0015] Furthermore, it can also include any kind of cosmetic excipient base; Cosmetic excipient bases include one or more of the following: solvents, moisturizers, chelating agents, emollients, skin conditioning agents, emulsifiers, fragrances, preservatives, thickeners, and pH adjusters.

[0016] The present invention also proposes a product with anti-aging function, the product comprising the above-mentioned short oligonucleotide, the above-mentioned double-stranded oligonucleotide and / or the above-mentioned short oligonucleotide composition.

[0017] Furthermore, the anti-aging function includes promoting SIRT-1 expression.

[0018] Furthermore, this also includes any cosmetic excipient base; Cosmetic excipient bases include one or more of the following: solvents, moisturizers, chelating agents, emollients, skin conditioning agents, emulsifiers, fragrances, preservatives, thickeners, and pH adjusters.

[0019] The technical solution of this invention has the following advantages: (1) The components are well-defined, and the batch stability and controllability are excellent. This invention uses chemically synthesized short oligonucleotides with fixed sequences, which is different from the random fragment mixture of traditionally extracted PDRN and DNA-sodium. The structure, length and base sequence of the raw materials are precisely controllable, which can realize sequence-level quality control. This effectively solves the problems of messy components, large batch differences and difficulty in standardization of natural nucleic acid raw materials, and greatly improves the batch consistency of products.

[0020] (2) Clear technical boundaries. By defining specific oligonucleotide sequences, fixing base pairing relationships and clearly defined efficacy application scenarios, this invention constructs a technical system in which structure and use are precisely matched, overcoming the defects of existing mixed nucleic acid systems with ambiguous structure and inability to define core functional units, thus forming a proprietary technical solution with clear boundaries and precise positioning.

[0021] (3) It has both whitening and anti-aging effects. Experimental results show that the relevant samples of this invention can promote SIRT-1 expression at a concentration of 20 μg / mL and inhibit melanin production at a concentration of 100 μg / mL, which can provide experimental basis for the development of skin whitening and anti-aging related products.

[0022] (4) The whitening effect of some samples is superior to that of traditional PDRN. Parallel comparative experiments confirmed that, under the experimental conditions of 100 μg / mL, the melanin inhibition effect of some samples of the present invention is significantly better than that of commercial PDRN raw materials. Among them, the relative melanin content of sample YH05F+YH05R was 51.023±2.336, which was significantly lower than that of the PDRN control group of 81.163±2.222, indicating that this short oligonucleotide pairing system has good potential for whitening applications.

[0023] (5) The process is highly standardized, safe, and easy to industrialize. This invention uses a chemical synthesis process, which can effectively avoid the safety hazards of protein residues, endotoxins, and impurities present in natural extraction processes. The purity and impurity content of raw materials are controllable and the quality is traceable, resulting in higher safety. At the same time, the chemical synthesis process has good repeatability and a high degree of standardization, making it suitable for large-scale and industrialized production and application of cosmetic raw materials. Attached Figure Description

[0024] To more clearly illustrate the specific embodiments of the present invention or the technical solutions in the prior art, the drawings used in the description of the specific embodiments or the prior art will be briefly introduced below. Obviously, the drawings described below are some embodiments of the present invention. For those skilled in the art, other drawings can be obtained from these drawings without creative effort.

[0025] Figure 1 This is a graph showing the cytotoxicity results of short oligonucleotide samples from this invention; Figure 2 This is a graph showing the results of the short oligonucleotide sample promoting SIRT-1 expression in this invention; Figure 3 This is a diagram showing the results of inhibiting melanin production in short oligonucleotide samples according to the present invention. Detailed Implementation

[0026] The following embodiments are provided to better understand the present invention, but the following embodiments do not constitute a limitation on the content and scope of protection of the present invention. Any product that is the same as or similar to the present invention, derived by any person under the guidance of the present invention or by combining the features of the present invention with other prior art, falls within the scope of protection of the present invention.

[0027] Unless otherwise specified, all experimental steps or conditions in the examples were performed according to conventional experimental procedures and conditions in the art. Reagents or instruments whose manufacturers are not specified are all commercially available products.

[0028] Furthermore, regarding the numerical ranges in this invention, it should be understood that each intermediate value between the upper and lower limits of the range is also specifically disclosed. Any stated value or intermediate value within a stated range, as well as each smaller range between any other stated value or intermediate value within said range, are also included in this invention. The upper and lower limits of these smaller ranges may be independently included or excluded from the range.

[0029] Unless otherwise stated, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art. While only preferred methods and materials have been described herein, any methods and materials similar to or equivalent to those described herein may be used in the implementation or testing of this invention.

[0030] The terms “include,” “including,” “have,” “contain,” etc., used in this article are all open-ended terms, meaning that they include but are not limited to.

[0031] This invention provides a chemically synthesized short oligonucleotide derived from the salmon genome and its application in whitening and / or anti-aging cosmetics, aiming to solve the technical defects of existing natural nucleic acid raw materials, such as uncontrollable components, single efficacy, unclear structure-efficacy correspondence, and poor safety.

[0032] First, this invention proposes a short oligonucleotide, the nucleotide sequence of which is shown in any one of SEQ ID NO:1 to SEQ ID NO:14. This short oligonucleotide can be used to form double-stranded oligonucleotides or short oligonucleotide compositions with whitening and / or anti-aging related functions. These short oligonucleotides are chemically synthesized short oligonucleotides derived from the salmon genome.

[0033] This invention also proposes a double-stranded oligonucleotide with whitening and / or anti-aging functions formed from the above-mentioned short oligonucleotides, including at least one of the complementary strand combinations YH01, YH02, YH03, YH04, YH05, YH06, and YH07. Further, in the complementary strand combinations YH01, YH02, YH03, YH04, YH05, YH06, and YH07, the sense and antisense strands are mixed in an equimolar ratio.

[0034] The aforementioned short oligonucleotides can form an equimolar mixture of complementary strands. This equimolar mixture can be annealed to form double-stranded oligonucleotides, or it can be used as a complementary single-stranded mixture without specific annealing. The annealing process involves placing the equimolar mixture in a common annealing buffer and heating at 95°C for 5-10 min to fully denature the nucleic acid single strands; subsequently, it is slowly cooled to room temperature at a rate of 1-2°C / min to promote annealing and pairing of the two complementary strands.

[0035] The present invention also proposes a short oligonucleotide composition containing the above-mentioned short oligonucleotides or double-stranded oligonucleotides and having whitening and / or anti-aging functions.

[0036] This invention also proposes products with whitening functions using the above-mentioned short oligonucleotides, the above-mentioned double-stranded oligonucleotides, and / or combinations of the above-mentioned short oligonucleotides as active ingredients. The whitening function includes inhibiting melanin production. Experimental verification shows that the melanin inhibition effect of some samples of this invention is significantly better than that of commercially available PDRN raw materials. Specifically, the relative melanin content of sample YH05F+YH05R was 51.023±2.336, significantly lower than the 81.163±2.222 of the PDRN control group, indicating that this short oligonucleotide pairing system has good potential for whitening applications.

[0037] This invention also proposes products with anti-aging functions using the aforementioned short oligonucleotides, double-stranded oligonucleotides, and / or combinations of the aforementioned short oligonucleotides as active ingredients. The anti-aging function includes promoting SIRT-1 expression. Experimental verification shows that the samples of this invention can promote SIRT-1 expression at a concentration of 20 μg / mL and inhibit melanin production at a concentration of 100 μg / mL, providing experimental evidence for the development of skin whitening, brightening, and anti-aging related products.

[0038] The aforementioned products may also include any cosmetic excipient base; cosmetic excipient bases include one or more of the following: solvents, moisturizers, chelating agents, emollients, skin conditioning agents, emulsifiers, fragrances, preservatives, thickeners, and pH adjusters.

[0039] Explanation of terms used in this application: Oligonucleotides: short single-stranded DNA fragments synthesized artificially by chemical means; Equimolar mixture: A system formed by mixing two complementary single chains at the same molar concentration (unannealed); Annealed double-stranded DNA: A stable double-stranded structure formed by two complementary single strands being denatured by heating and then slowly cooled to pair up; SIRT-1: Silent Information Regulator 1, is a key protein that regulates cellular anti-aging and photodamage repair; PDRN: Polydeoxyribonucleotide, usually a mixture of DNA fragments extracted from salmon tissue; B16-F10 cells: mouse melanoma cells used to evaluate whitening / melanin-inhibiting efficacy; HFF-1 cells: human skin fibroblasts, used to evaluate anti-aging and repair effects; HaCaT cells: human keratinocytes, used to evaluate the safety of cosmetic ingredients.

[0040] Various exemplary embodiments of the present invention will now be described in detail. This detailed description should not be considered as a limitation of the present invention, but rather as a more detailed description of certain aspects, features, and embodiments of the present invention. It should be understood that the terminology used in this invention is merely for describing particular embodiments and is not intended to limit the present invention.

[0041] Example 1. Preparation of short oligonucleotides derived from salmon genome This embodiment aims to prepare short oligonucleotides derived from the salmon genome. The specific steps are as follows: 1. Sequence selection The VGLL3 gene, a precocity control gene in Atlantic salmon (Salmo salar), was used for sequencing to prepare short oligonucleotide sequences. The screening process is as follows: First, candidate fragments were extracted from the relevant coding region of the gene, and molecular docking scoring was performed using HADDOCK3 software. The top-scoring sequences were selected as candidates. The specific screening criteria were as follows: 1. The sequence length was limited to 16-22 bp; 2. Sequences with obvious hairpin structures, self-aggregation effects, and other secondary structures that were not conducive to application were removed; 3. Sequences that highly overlapped with existing publicly available nucleic acid cosmetic sequence combinations were excluded to ensure the novelty and specificity of the sequence combinations. Seven pairs of 14 specific short oligonucleotide sequences (SEQ ID NO: 1-14, as shown in Table 1) were obtained, with sequence lengths of 16 bp, 17 bp, and 22 bp, respectively. Among them, YH01F and YH01R are complementary single-stranded pairs, YH02F and YH02R are complementary single-stranded pairs, YH03F and YH03R are complementary single-stranded pairs, YH04F and YH04R are complementary single-stranded pairs, YH05F and YH05R are complementary single-stranded pairs, YH06F and YH06R are complementary single-stranded pairs, and YH07F and YH07R are complementary single-stranded pairs. All oligonucleotides were chemically synthesized using the phosphoramide method and purified (synthesis and purification steps were outsourced to General Biotechnology Co., Ltd.). This preparation process ensures precise and well-defined oligonucleotide sequence structures, excellent batch-to-batch quality consistency, and effective control of product purity and various impurity indicators, achieving standardized and stable control of raw material quality.

[0042] Table 1.14 Specific Short Oligonucleotide Sequences

[0043] 2. Preparation of double-stranded DNA Fourteen specific short oligonucleotide sequences were dissolved in buffer solutions (10 mM Tris-HCl, 50 mM NaCl) at pH 7.5–8.0 to prepare oligonucleotide single-strand stock solutions with a final concentration of 100 μg / mL. Complementary oligonucleotide single-strand stock solutions were selected and, after molar concentration conversion, mixed in equimolar ratios. The mixtures were vortexed for approximately 10 seconds and allowed to stand at room temperature for approximately 3–10 minutes to obtain equimolar mixtures without special annealing treatment (referred to as YH01F+YH01R, YH02F+YH02R, YH03F+YH03R, YH04F+YH04R, YH05F+YH05R, YH06F+YH06R, and YH07F+YH07R).

[0044] An equimolar mixture without special annealing treatment was heated at 95°C for 5-10 min to fully denature the single-stranded nucleic acid. Then, it was slowly cooled to room temperature at a rate of 1-2°C / min to induce the complementary strands to complete annealing and pairing, thus obtaining a double-stranded sample. The double-stranded samples include: YH01F and YH01R forming a complementary pair after annealing, abbreviated as YH01; YH02F and YH02R forming a complementary pair after annealing, abbreviated as YH02; YH03F and YH03R forming a complementary pair after annealing, abbreviated as YH03; YH04F and YH04R forming a complementary pair after annealing, abbreviated as YH04; YH05F and YH05R forming a complementary pair after annealing, abbreviated as YH05; YH06F and YH06R forming a complementary pair after annealing, abbreviated as YH06; and YH07F and YH07R forming a complementary pair after annealing, abbreviated as YH07.

[0045] Experimental Example 1. Efficacy and Safety Verification 1. Cytotoxicity detection Human fibroblast toxicity assays were used to detect changes in cell viability. HFF-1 fibroblasts were used as the experimental model. Five concentrations of the following compounds were added: YH01, YH02, YH03, YH04, YH05, YH06, YH07, YH01F+YH01R, YH02F+YH02R, YH03F+YH03R, YH04F+YH04R, YH05F+YH05R, YH06F+YH06R, YH07F+YH07R, PDRN (purchased from Regmen), and sodium DNA (purchased from Regmen). Cell viability was assessed after 24 hours of treatment.

[0046] Within the concentration range of 0.16-100 μg / mL, the annealed double-stranded samples YH01-YH07, as well as the equimolar mixtures YH01F+YH01R, YH02F+YH02R, YH03F+YH03R, YH04F+YH04R, YH05F+YH05R, YH06F+YH06R, and YH07F+YH07R without special annealing treatment, all obtained complete corresponding cytotoxicity test results. The results are shown in Tables 2-9. Figure 1 The detection values ​​for each sample at different concentrations generally ranged from 81.206 to 104.335. Specifically, in the annealed double-stranded samples, the detection value for YH03 at a concentration of 100 μg / mL was 81.206 ± 1.729 (n=6), and the detection value for YH05 at a concentration of 100 μg / mL was 83.797 ± 1.836 (n=6). In the equimolar mixtures without special annealing treatment, the detection values ​​for YH01F+YH01R were 91.081-95.688 within the concentration range of 0.16-100 μg / mL, the corresponding detection values ​​for YH02F+YH02R were 91.031-97.957, and the corresponding detection values ​​for YH05F+YH05R were 94.036-97.404. These cytotoxicity test results can serve as the basis for evaluating the in vitro safety of the samples related to this invention.

[0047] Table 2. Cytotoxicity results of YH01 and YH02

[0048] Table 3. Cytotoxicity results of YH03 and YH04

[0049] Table 4. Cytotoxicity results of YH05 and YH06

[0050] Table 5. Cytotoxicity results of YH07, YH01F+YH01R

[0051] Table 6. Cytotoxicity results of YH02F+YH02R and YH03F+YH03R

[0052] Table 7. Cytotoxicity results of YH04F+YH04R and YH05F+YH05R

[0053] Table 8. Cytotoxicity results of YH06F+YH06R and YH07F+YH07R

[0054] Table 9. Cytotoxicity results of PDRN and DNA-sodium

[0055] 2. Effects of promoting SIRT-1 expression To evaluate the anti-aging efficacy of the test substance, this study used human fibroblasts as a model and detected the expression level of SIRT-1 in the cells by immunofluorescence, thus providing experimental evidence for efficacy evaluation.

[0056] The specific methods are as follows: Commercial PDRN (purchased from Regmen), DNA-sodium (purchased from Regmen), and resveratrol (purchased from Maclean) were used as positive controls. Cells were treated with 20 μg / mL of YH01, YH02, YH03, YH04, YH05, YH06, YH07, YH01F+YH01R, YH02F+YH02R, YH03F+YH03R, YH04F+YH04R, YH05F+YH05R, YH06F+YH06R, and YH07F+YH07R, respectively. The average intensity of immunofluorescence was used as the evaluation index to detect the promoting effect of each sample on SIRT-1 expression.

[0057] According to Table 10 and Figure 2 Among them, YH01-YH07 were double-stranded nucleic acid samples prepared by annealing; YH01F+YH01R, YH02F+YH02R, YH03F+YH03R, YH04F+YH04R, YH05F+YH05R, YH06F+YH06R, and YH07F+YH07R were equimolar mixtures of raw materials that had not undergone specific annealing treatment. The experimental results showed that at an effective concentration of 20 μg / mL, both the annealed double-stranded samples and some equimolar mixtures that had not undergone specific annealing exhibited significant SIRT-1 expression-promoting effects. Based on the mean detection results, samples YH05, YH03, YH05F+YH05R, and YH07 showed more pronounced SIRT-1 promoting effects.

[0058] Table 10. Effects on promoting SIRT-1 expression

[0059] 3. Inhibition of melanin production To evaluate the skin whitening efficacy of the tested samples, this study used B16-F10 melanocytes as a model and analyzed the changes in melanin content in the cells.

[0060] Using commercially available PDRN (purchased from Regmin), DNA-sodium (purchased from Regmin), and kojic acid (purchased from Aladdin) as controls, cells were treated with YH01, YH02, YH03, YH04, YH05, YH06, YH07, YH01F+YH01R, YH02F+YH02R, YH03F+YH03R, YH04F+YH04R, YH05F+YH05R, YH06F+YH06R, and YH07F+YH07R, respectively. The inhibitory effect of each sample on melanin production was then detected, and the results are shown in Table 11. Among them, YH01~YH07 are double-chain samples that have undergone annealing treatment, while YH01F+YH01R, YH02F+YH02R, YH03F+YH03R, YH04F+YH04R, YH05F+YH05R, YH06F+YH06R and YH07F+YH07R are equimolar mixed samples that have not been specially annealed.

[0061] The experimental results are shown in Table 11 and Figure 3 As shown in the figure, YH01–YH07 are annealed double-stranded samples, while YH01F+YH01R, YH02F+YH02R, YH03F+YH03R, YH04F+YH04R, YH05F+YH05R, YH06F+YH06R, and YH07F+YH07R are equimolar mixtures without specific annealing treatment. Experimental results show that at a concentration of 100 μg / mL, both the annealed double-stranded samples and some of the equimolar mixtures without specific annealing significantly inhibit melanin production. The mean values ​​show that samples YH06F+YH06R, YH05F+YH05R, YH04F+YH04R, YH06, YH05, and YH04 exhibit superior inhibition effects. Among them, the mean value of YH04F+YH04R under the experimental conditions was 41.463 ± 0.222, which was significantly lower than the 81.163 ± 2.222 of the PDRN control group.

[0062] Table 11. Results of melanin production inhibition

[0063] In summary, based on the above-mentioned cytotoxicity, SIRT-1 expression promotion, and melanin production inhibition experimental results, it can be concluded that the annealed double-stranded samples and some equimolar mixtures without special annealing treatment involved in this invention possess the following technical effects under the same or similar test conditions: firstly, corresponding basic data for in vitro safety evaluation were obtained; secondly, SIRT-1 expression was promoted at a concentration of 20 μg / mL; and thirdly, SIRT-1 expression was promoted at a concentration of 100 μg / mL. At a concentration of μg / mL, melanin production can be inhibited. Among them, samples YH05, YH03, YH05F+YH05R, and YH07 showed better effects in promoting SIRT-1 expression, while samples YH06F+YH06R, YH05F+YH05R, YH04F+YH04R, YH06, YH05, and YH04 showed good effects in inhibiting melanin production. The cytotoxicity test results showed that YH01F+YH01R, YH02F+YH02R, and YH05F+YH05R all obtained complete and reliable in vitro safety data within the tested concentration range. At the same time, PDRN was set as a control group in this study, which can provide experimental support for subsequent efficacy comparison with existing PDRN raw materials.

[0064] Obviously, the above embodiments are merely illustrative examples for clear explanation and are not intended to limit the implementation. Those skilled in the art will recognize that other variations or modifications can be made based on the above description. It is neither necessary nor possible to exhaustively list all possible implementations here. However, obvious variations or modifications derived therefrom are still within the scope of protection of this invention.

Claims

1. A short oligonucleotide, characterized in that, The nucleotide sequence of the short oligonucleotide is shown in any one of SEQ ID NO:1 to SEQ ID NO:

14.

2. A double-stranded oligonucleotide with whitening and / or anti-aging functions, characterized in that, Including at least one of the complementary chain combinations YH01, YH02, YH03, YH04, YH05, YH06, and YH07; The nucleotide sequence of the sense strand in the complementary strand combination YH01 is shown in SEQ ID NO:1, and the nucleotide sequence of the antisense strand is shown in SEQ ID NO:

2. The nucleotide sequence of the sense strand of the complementary strand combination YH02 is shown in SEQ ID NO:3, and the nucleotide sequence of the antisense strand is shown in SEQ ID NO:

4. The nucleotide sequence of the sense strand of the complementary strand combination YH03 is shown in SEQ ID NO:5, and the nucleotide sequence of the antisense strand is shown in SEQ ID NO:

6. The nucleotide sequence of the sense strand of the complementary strand combination YH04 is shown in SEQ ID NO:7, and the nucleotide sequence of the antisense strand is shown in SEQ ID NO:

8. The nucleotide sequence of the sense strand of the complementary strand assembly YH05 is shown in SEQ ID NO:9, and the nucleotide sequence of the antisense strand is shown in SEQ ID NO:

10. The nucleotide sequence of the sense strand of the complementary strand combination YH06 is shown in SEQ ID NO:11, and the nucleotide sequence of the antisense strand is shown in SEQ ID NO:

12. The nucleotide sequence of the sense strand of the complementary strand assembly YH07 is shown in SEQ ID NO:13, and the nucleotide sequence of the antisense strand is shown in SEQ ID NO:

14.

3. A short oligonucleotide composition having whitening and / or anti-aging functions, characterized in that, The short oligonucleotide composition comprises the short oligonucleotide of claim 1 and / or the double-stranded oligonucleotide with whitening and / or anti-aging functions of claim 2.

4. The double-stranded oligonucleotide with whitening and / or anti-aging function according to claim 2 or the short oligonucleotide composition with whitening and / or anti-aging function according to claim 3, wherein in the complementary chain combination YH01, YH02, YH03, YH04, YH05, YH06, YH07, the sense chain and antisense chain are mixed in an equimolar ratio; and / or, the complementary chain combination is subjected to annealing treatment.

5. A product with whitening function, characterized in that, The product comprises the short oligonucleotide of claim 1, the double-stranded oligonucleotide of claim 2, and / or the short oligonucleotide composition of claim 3.

6. The product according to claim 5, characterized in that, The product also includes cosmetic excipient base.

7. The product according to claim 6, characterized in that, The cosmetic excipient matrix includes one or more of the following: solvent, moisturizer, chelating agent, emollient, skin conditioning agent, emulsifier, fragrance, preservative, thickener, and pH adjuster.

8. A product with anti-aging function, characterized in that, The product comprises the short oligonucleotide of claim 1, the double-stranded oligonucleotide of claim 2, and / or the short oligonucleotide composition of claim 3.

9. The product according to claim 8, characterized in that, The product also includes cosmetic excipient base.

10. The product according to claim 9, characterized in that, The cosmetic excipient matrix includes one or more of the following: solvent, moisturizer, chelating agent, emollient, skin conditioning agent, emulsifier, fragrance, preservative, thickener, and pH adjuster.

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