Tetrapeptide for inhibiting melanogenesis as well as composition and application thereof
Through the combination of tetrapeptide CHWH designed by bioinformatics, combined with nicotinamide and α-arbutin, the inhibition of melanin production of multiple targets and multiple pathways is achieved, and the single pathway and side effects of existing whitening products in inhibiting melanin production are solved. Problem, significantly improving the inhibitory effect and safety.
Patent Information
- Application Number
- CN202411751808.3
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2024-12-02
- Publication Date
- 2025-02-14
AI Technical Summary
Existing whitening cosmetics have a single pathway intervention in inhibiting melanin production, with great side effects and limited effects, making it difficult to achieve comprehensive melanin production inhibition.
Through bioinformatics design, a tetrapeptide CHWH that inhibits melanin production was developed and combined with nicotinamide and α-arbutin to synergize to inhibit melanin production in a multi-target and multi-pathway manner.
It achieves lower cytotoxicity and better stability, can effectively inhibit melanin production, achieve higher inhibitory effect at lower concentrations than a single active raw material, and has better safety.
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Figure CN119431494A_ABST
Abstract
Description
Technical Field
[0001] The present invention relates to the technical field of cosmetics, and relates to a tetrapeptide for inhibiting melanin production, a composition thereof and an application thereof, and in particular to an active tetrapeptide CHWH designed by bioinformatics, a composition of the active tetrapeptide CHWH, nicotinamide and α-arbutin for synergistically inhibiting melanin production and an application thereof. Background Art
[0002] With the improvement of material living standards, people have higher and higher requirements for life, pay more and more attention to "appearance beauty", and the demand for whitening cosmetics is also increasing day by day. Whitening cosmetics with different ingredients and mechanisms continue to appear in the market, and the speed of replacement is very rapid.
[0003] According to the mechanism of action, cosmetics with whitening effects on the market can be divided into physical covering and chemical whitening according to the form of their action. Physical covering products use physical covering agents, such as titanium dioxide, zinc oxide, talcum powder, etc., which are applied to the skin surface to cover the spots on the skin to achieve a whitening effect, but they cannot completely change the original appearance of the skin. Long-term use will cause blockage of pores or sebaceous glands, affect sweating and metabolism, and lead to skin problems such as acne. Chemical whitening products achieve the effect of whitening the skin by inhibiting the production of melanin, blocking the transportation of melanin, and accelerating the metabolism of epidermal cells. With the continuous deepening of research on melanin synthesis, it is found that the formation of melanin mainly involves three enzymes: tyrosinase (TYR), DHICA oxidase (TRP-1) and dopachrome isomerase (TRP2), among which tyrosinase is the most critical rate-limiting enzyme in the process of melanin production.
[0004] Currently, whitening products on the market mainly achieve whitening effects by reducing melanin production, hindering melanin transport, and promoting melanin metabolism. Many whitening active ingredients have been discovered, including kojic acid, phenylethyl resorcinol, vitamin C, tranexamic acid, niacinamide, α-arbutin, etc. However, studies have shown that most of these ingredients act on a relatively single pathway, inhibiting melanin production or controlling the transport from melanosomes to the stratum corneum by intervening in a certain target. For example, vitamin C can reduce the generated melanin; niacinamide can block the transfer of melanin from epidermal cells to the stratum corneum, and promote melanin metabolism by accelerating the renewal rate of skin cells; α-arbutin reversibly inhibits tyrosinase activity through a mixed inhibition of competitive and non-competitive inhibition; kojic acid can act as the active center of tyrosinase CU. 2+ Complexing agent, etc.
[0005] Compared with traditional drugs, biological small molecule active peptides have the advantages of high efficacy, short sequence, low toxicity, and high cell permeability. Due to the different side chains carried by different amino acids in the amino acid sequence, the active peptides can be effective on multiple pathways and multiple targets. Compared with traditional active ingredients, small molecule active peptides have significantly fewer side effects such as damage and sensitization to the body, and have the significant advantage of high efficiency and low toxicity.
[0006] Therefore, it is of great practical significance to develop a biological small molecule active peptide that can effectively inhibit the production of melanin. Summary of the invention
[0007] Due to the above-mentioned defects in the prior art, the present invention provides a small molecule biological active peptide that can effectively inhibit the production of melanin, specifically a tetrapeptide that inhibits the production of melanin, a composition thereof and an application thereof.
[0008] In order to achieve the above object, the present invention provides the following technical solutions:
[0009] A tetrapeptide for inhibiting melanin production, whose amino acid sequence is Cys-His-Trp-His.
[0010] Compared with traditional drugs, biological small molecule peptides have the advantages of high efficiency, short sequence, low toxicity, high cell permeability, etc. The above-mentioned tetrapeptide that inhibits melanin production has been tested to be effective. It can not only reduce the production of melanin by irreversible inhibition, but also reduce melanin through strong antioxidant properties, and can act as a chelating agent with the CU at the active center of tyrosinase. 2+ Complexation changes the spatial structure of the enzyme to reduce the activity of tyrosinase. In addition, the above tetrapeptide that inhibits melanin production has good stability and safety.
[0011] As the preferred technical solution:
[0012] As described above, the tetrapeptide that inhibits melanin production, the confirmation process of the tetrapeptide that inhibits melanin production is: first, a model is constructed by homology modeling, and screening is performed by bioinformatics methods to analyze the action mode and binding site of the active tetrapeptide with the receptor protein, and the peptide with the best activity is the tetrapeptide that inhibits melanin production.
[0013] A tetrapeptide for inhibiting melanin production as described above, wherein the bioinformatics method is molecular docking;
[0014] The analysis of the action mode and binding site of the active tetrapeptide and the receptor protein was performed using PYMOL software.
[0015] The present invention provides a method for preparing the tetrapeptide for inhibiting melanin production as described above, wherein the tetrapeptide for inhibiting melanin production is prepared by solid phase synthesis.
[0016] As the preferred technical solution:
[0017] The method as described above comprises the following steps:
[0018] S1. Activate WANG resin with three volumes of DCM (dichloromethane);
[0019] S2. Accurately weigh the required His, fully activate it in an ice bath, connect His to WANG resin using DMAP (4-dimethylaminopyridine) / DIC (N,N'-diisopropylcarbodiimide) / HOBT (1-hydroxybenzotriazole) system, and use the ninhydrin method to detect the connection status;
[0020] S3, washing the resin: wash the resin three times with three volumes of DMF (N,N-dimethylformamide) / DCM / MEOH (methanol);
[0021] S4, capping the WANG resin with an acetic anhydride:pyridine system, and washing the resin three times with three volumes of DMF / DCM / MEOH;
[0022] S5, using 20% piperidine solution for deprotection, the deprotection operation is performed twice, and the resin is washed three times with three volumes of DMF / DCM / MEOH, wherein the solvent of the piperidine solution is DMF;
[0023] S6, repeat the steps of S2, S3, and S5 to connect Trp, His, and Cys in sequence;
[0024] S7. Cut the resin using TFA (trifluoroacetic acid) / thioanisole / phenol / water system, cut the connected peptide chains and freeze-dry them to obtain the tetrapeptide that inhibits melanin production.
[0025] The purity of the tetrapeptide for inhibiting melanin production obtained by the method described above reaches more than 98%.
[0026] The present invention also provides a composition with the effect of inhibiting melanin production, comprising tetrapeptide for inhibiting melanin production, nicotinamide and α-arbutin.
[0027] The above-mentioned tetrapeptide that inhibits melanin production is combined with niacinamide and α-arbutin to achieve synergistic effects, and multiple targets work together to achieve the effect of multi-pathway inhibition of melanin production and ensure the safety of the combination system.
[0028] As the preferred technical solution:
[0029] The composition having the effect of inhibiting melanin production as described above further comprises deionized water;
[0030] The mass proportions of the tetrapeptide, nicotinamide and α-arbutin for inhibiting melanin production in the composition are 0.03-0.05wt%, 0.3-0.5wt% and 0.1-0.3wt% respectively.
[0031] In addition, the present invention also provides a method for preparing the above-mentioned composition having the effect of inhibiting melanin production, wherein the tetrapeptide, nicotinamide and α-arbutin that inhibit melanin production are mixed in corresponding proportions, deionized water is added and stirred intermittently to obtain the composition having the effect of inhibiting melanin production, wherein the stirring speed of the intermittent stirring is 200-400 rpm, and the stirring time is 15-20 min.
[0032] Furthermore, the present invention also provides the use of the above-mentioned tetrapeptide for inhibiting melanin production and the above-mentioned composition having the effect of inhibiting melanin production in the preparation of cosmetics.
[0033] The above technical solution is only a feasible technical solution of the present invention, and the protection scope of the present invention is not limited thereto. Those skilled in the art can reasonably adjust the specific design according to actual needs.
[0034] The above invention has the following advantages or beneficial effects:
[0035] (1) Compared with traditional raw materials, the tetrapeptide of the present invention has lower cytotoxicity and better stability. It reduces melanin by virtue of its strong reducing power and can irreversibly inhibit the activity of tyrosinase. It also acts as a complexing agent to complex the double CU in the active center of tyrosinase. 2+ , changing the spatial structure of the enzyme to reduce tyrosinase activity;
[0036] (2) The composition of the present invention having the effect of inhibiting melanin production, wherein niacinamide in the composition blocks the transfer of melanin from epidermal cells to the stratum corneum, promotes melanin metabolism by accelerating the renewal rate of skin cells, and α-arbutin inhibits tyrosinase activity by a mixed inhibition of competitive and non-competitive inhibition;
[0037] (3) Compared with a single active raw material, the composition of the present invention having the effect of inhibiting melanin production can achieve a higher melanin inhibition effect at a lower concentration, and at the same time has better safety and good application prospects. BRIEF DESCRIPTION OF THE DRAWINGS
[0038] The present invention and its features, appearance and advantages will become more apparent by reading the detailed description of non-limiting embodiments with reference to the following drawings. The same reference numerals indicate the same parts in all the drawings. The drawings are not drawn to scale, and emphasis is placed on illustrating the subject matter of the present invention.
[0039] Figure 1 It is the liquid phase diagram of the solid phase synthesis of tetrapeptide CHWH;
[0040] Figure 2 This is the mass spectrum of the solid phase synthesized tetrapeptide CHWH;
[0041] Figure 3 The DPPH clearance rate and ABTS clearance rate of tetrapeptide CHWH at different concentrations + Positive ion clearance and CU 2+ Schematic diagram of the effect of chelation rate;
[0042] Figure 4 Schematic diagram of the effect of different concentrations of tetrapeptide CHWH on B16 cell viability;
[0043] Figure 5 Schematic diagram of the effect of different concentrations of tetrapeptide CHWH on the inhibition rate of B16 tyrosinase activity and melanin production;
[0044] Figure 6 It is a comparative schematic diagram of the B16 cell viability of Examples 1 to 9 and Comparative Examples 1 to 5;
[0045] Figure 7 It is a comparative diagram of the inhibition rate of tyrosinase activity and melanin synthesis in B16 cells of Examples 1 to 9 and Comparative Examples 1 to 5. DETAILED DESCRIPTION
[0046] The structure of the present invention is further described below in conjunction with the accompanying drawings and specific embodiments, but is not intended to limit the present invention.
[0047] A method for preparing a tetrapeptide CHWH for inhibiting melanin production, which is obtained by solid phase synthesis, and the specific process is as follows:
[0048] S1, activate WANG resin with three volumes of DCM;
[0049] S2. Accurately weigh the first amino acid (His) required, fully activate it in an ice bath, connect the amino acid to the WANG resin using the DMAP / DIC / HOBT system, and detect the connection status using the ninhydrin method;
[0050] S3, washing resin: washing the resin three times with three volumes of DMF / DCM / MEOH;
[0051] S4, capping the WANG resin with an acetic anhydride:pyridine system, and washing the resin three times with three volumes of DMF / DCM / MEOH;
[0052] S5. Deprotection was performed using 20% piperidine solution (solvent: DMF), the deprotection operation was performed twice, and the resin was washed three times with three volumes of DMF / DCM / MEOH;
[0053] S6, repeat the operations of S2, S3, and S5, and connect the second (Trp), third (His), and fourth (Cys) amino acids in sequence (the synthesis direction is from C-terminus to N-terminus);
[0054] S7. Use TFA / thioanisole / phenol / water system to cut the resin, cut the connected peptide chains and freeze-dry them to obtain the tetrapeptide CHWH that inhibits melanin production.
[0055] The liquid phase diagram and mass spectrum of the prepared tetrapeptide CHWH are as follows: Figure 1 and Figure 2 shown.
[0056] An embodiment of the present invention provides a composition having the effect of inhibiting melanin production, the composition comprising: tetrapeptide CHWH for inhibiting melanin production, nicotinamide, α-arbutin and deionized water; wherein the amino acid sequence of tetrapeptide CHWH for inhibiting melanin production is: Cys-His-Trp-His.
[0057] The following composition preparation method is as follows: tetrapeptide CHWH, nicotinamide, and α-arbutin are mixed in the corresponding proportions, the balance is supplemented with deionized water, and intermittent stirring is performed (200-400 rpm, stirring for 15-20 minutes) to ensure the formation of a stable and uniform system.
[0058] The composition utilizes different action pathways and mechanisms of different components to achieve a comprehensive melanin inhibition effect.
[0059] The present invention will be further explained below by means of specific examples.
[0060] Example 1: In terms of mass fraction, 0.03wt% tetrapeptide CHWH, 0.3wt% nicotinamide, and 0.1wt% α-arbutin were mixed in corresponding proportions, deionized water was added, and the mixture was stirred intermittently (200rpm / min, stirring for 15min) to form a stable and uniform system.
[0061] Example 2: In terms of mass fraction, 0.03wt% tetrapeptide CHWH, 0.4wt% nicotinamide, and 0.2wt% α-arbutin were mixed in corresponding proportions, deionized water was added, and the mixture was stirred intermittently (300rpm / min, stirring for 18min) to form a stable and uniform system.
[0062] Example 3: In terms of mass fraction, 0.03wt% tetrapeptide CHWH, 0.5wt% nicotinamide, and 0.3wt% α-arbutin were mixed in corresponding proportions, deionized water was added, and the mixture was stirred intermittently (400rpm / min, stirring for 20min) to form a stable and uniform system.
[0063] Example 4: In terms of mass fraction, 0.04wt% tetrapeptide CHWH, 0.3wt% niacinamide, and 0.1wt% α-arbutin were mixed in corresponding proportions, deionized water was added, and the mixture was stirred intermittently (200rpm / min, stirring for 15min) to form a stable and uniform system.
[0064] Example 5: In terms of mass fraction, 0.04wt% tetrapeptide CHWH, 0.4wt% nicotinamide, and 0.2wt% α-arbutin were mixed in corresponding proportions, deionized water was added, and the mixture was stirred intermittently (300rpm / min, stirring for 18min) to form a stable and uniform system.
[0065] Example 6: In terms of mass fraction, 0.04wt% tetrapeptide CHWH, 0.5wt% niacinamide, and 0.3wt% α-arbutin were mixed in corresponding proportions, deionized water was added, and the mixture was stirred intermittently (400rpm / min, stirring for 20min) to form a stable and uniform system.
[0066] Example 7: In terms of mass fraction, 0.05wt% tetrapeptide CHWH, 0.3wt% niacinamide, and 0.1wt% α-arbutin were mixed in corresponding proportions, deionized water was added, and the mixture was stirred intermittently (200rpm / min, stirring for 15min) to form a stable and uniform system.
[0067] Example 8: In terms of mass fraction, 0.05wt% tetrapeptide CHWH, 0.4wt% niacinamide, and 0.2wt% α-arbutin were mixed in corresponding proportions, deionized water was added, and the mixture was stirred intermittently (300rpm / min, stirring for 18min) to form a stable and uniform system.
[0068] Example 9: In terms of mass fraction, 0.05wt% tetrapeptide CHWH, 0.5wt% niacinamide, and 0.3wt% α-arbutin were mixed in corresponding proportions, deionized water was added, and the mixture was stirred intermittently (400rpm / min, stirring for 20min) to form a stable and uniform system.
[0069] The present invention is further described with reference to the following comparative examples:
[0070] Comparative Example 1: 0.3 wt % nicotinamide and 0.1 wt % α-arbutin were mixed in corresponding proportions, deionized water was added, and the mixture was stirred intermittently (200 rpm / min, stirring for 15 min) to form a stable and uniform system.
[0071] Comparative Example 2: 0.4wt% nicotinamide and 0.2wt% α-arbutin were mixed in corresponding proportions, deionized water was added, and the mixture was stirred intermittently (300rpm / min, stirring for 18min) to form a stable and uniform system.
[0072] Comparative Example 3: 0.5wt% nicotinamide and 0.3wt% α-arbutin were mixed in corresponding proportions, deionized water was added, and the mixture was stirred intermittently (400rpm / min, stirring for 20min) to form a stable and uniform system.
[0073] Comparative Example 4: 0.33 wt % nicotinamide and 0.1 wt % α-arbutin were mixed in corresponding proportions, deionized water was added, and the mixture was stirred intermittently (200 rpm / min, stirring for 15 min) to form a stable and uniform system.
[0074] Comparative Example 5: 0.44 wt% nicotinamide and 0.2 wt% α-arbutin were mixed in corresponding proportions, deionized water was added, and the mixture was stirred intermittently (300 rpm / min, stirring for 18 min) to form a stable and uniform system.
[0075] In order to further illustrate the efficacy and advantages of the composition, Examples 1 to 9 and Comparative Examples 1 to 5 were tested according to the following scheme:
[0076] Cytotoxicity assay
[0077] The experiment was carried out based on mouse B16 melanoma cells. The cells were cultured to the logarithmic phase using 1640 medium (10% FBS, 1% double antibody), and evenly plated in a 96-well plate at a density of 1*105 / well for culture. After 24 hours, 100ul of samples of different concentrations were added to each well. After 48 hours, the cell lethality was detected by the CCK-8 method, and the cell lethality of different embodiments and comparative examples was compared.
[0078] Experimental results: The CCK-8 method was used to determine the effects of Examples 1 to 9 and Comparative Examples 1 to 5 on the activity of mouse melanoma cells to determine the safe concentration range of the samples to be tested. Figure 6As shown, the effects of Examples 1 to 9 on the activity of B16 cells were not significant, and the cell viabilities were all above 90%. Among them, the cell viabilities of Examples 7 to 9 exceeded 100%, which had a certain promoting effect on cell proliferation. This may be due to the high concentration of tetrapeptide CHWH having an activating effect on B16 cell proliferation.
[0079] (1) Inhibition of tyrosinase activity
[0080] The experiment was carried out based on mouse B16 melanoma cells. The cells were cultured to the logarithmic phase with DMEM medium (10% FBS, 1% double antibody), and the cell suspension was prepared by counting with the trypan blue method. The cells were evenly plated on a 96-well plate at a density of 1*105 / well for culture. After 24 hours, the culture medium was discarded, and 100ul of culture medium containing the corresponding concentration (each embodiment and comparative example) was added to each well. After 48 hours of culture, the culture medium was discarded, and each well was lysed with 50ul of 1% Triton-100 solution, and then 50ul of L-DOPA solution was added for color development. The OD475nm value was detected by an enzyme reader, and the inhibition rate of B16 cell tyrosinase was calculated by colorimetry.
[0081] (2) Inhibition of melanin production
[0082] The experiment was conducted based on mouse B16 melanoma cells. The cells were cultured to the logarithmic phase using DMEM culture medium (10% FBS, 1% double antibody), and the cell suspension was prepared by counting with the trypan blue method. The cells were evenly plated in a 24-well plate at a density of 1*104 / well for culture. The culture medium was discarded after 24 hours, and 1 ml of culture medium containing the corresponding concentration (each embodiment and comparative example) was added to each well. After 48 hours of culture, the culture medium was discarded, and 500 ul of trypsin was added to each well for digestion for 3 minutes. Then, 1.5 ml of DMEM culture medium (10% FBS, 1% double antibody) was used to terminate the digestion. The cell suspension was collected and centrifuged, and the cells were lysed with NAOH to release melanin. The OD475nm value was detected by an enzyme reader, and the inhibition of B16 cell melanogenesis was calculated by colorimetry.
[0083] The experimental results are as follows Figure 7 As shown: Examples 1 to 9 have good inhibitory effects on B16 cell tyrosinase activity and melanin synthesis, can well inhibit melanin production, and the overall inhibitory effect of the examples is significantly better than that of the comparative example (the comparative example is a combination without tetrapeptide CHWH), among which Examples 7 to 9 have the most significant inhibitory effect. Examples 7 to 9 all contain a relatively high concentration of 0.05wt% tetrapeptide CHWH, and when the concentrations of nicotinamide and α-arbutin are the same, with the increase of the tetrapeptide content, the melanin synthesis inhibition rate and the tyrosinase activity inhibition rate are significantly improved, which shows that the tetrapeptide CHWH plays an important role in inhibiting melanin synthesis. And the composition with the addition of tetrapeptide CHWH can enable the system to achieve better inhibitory effects at a lower concentration, thereby ensuring that the composition system is more safe for the body.
[0084] Effects of different concentrations of tetrapeptide CHWH on DPPH clearance and ABTS + Positive ion clearance and CU 2+ The effect of chelation rate is Figure 3 As shown in the figure, the tetrapeptide CHWH has good antioxidant activity. At a low concentration of 0.125 mg / ml, its DPPH free radical scavenging rate can reach more than 70%, and the ABTS+ free radical scavenging rate can reach 80%. At a concentration of about 0.3 mg / ml, CU 2+ The chelation rate can reach 50%; the effects of different concentrations of tetrapeptide CHWH on B16 cell viability are as follows Figure 4 As shown in the figure, it can be seen that the tetrapeptide CHWH has no killing effect on B16 cells at low concentrations, and has a certain promoting effect on B16 cell activity at a high concentration of 0.5 mg / ml; the effects of different concentrations of tetrapeptide CHWH on the inhibition rate of B16 tyrosinase activity and melanin production are shown in Figure 5 As shown, the tetrapeptide CHWH has a relatively excellent B16 tyrosinase activity inhibition rate and melanin production inhibition rate. At a concentration of 0.18 mg / ml, the B16 tyrosinase activity inhibition rate reaches more than 50%, and at a concentration of 0.12 mg / ml, the B16 melanin production inhibition rate reaches 50%. At higher concentrations, it can significantly inhibit B16 tyrosinase activity and melanin production.
[0085] Those skilled in the art should understand that those skilled in the art can implement variations by combining the prior art and the above embodiments, which will not be described in detail here. Such variations do not affect the essential content of the present invention, and will not be described in detail here.
[0086] The above describes the preferred embodiments of the present invention. It should be understood that the present invention is not limited to the above-mentioned specific embodiments, and the devices and structures that are not described in detail should be understood to be implemented in a common manner in the art; any technician familiar with the art can use the above-disclosed methods and technical contents to make many possible changes and modifications to the technical solutions of the present invention without departing from the scope of the technical solutions of the present invention, or modify them into equivalent embodiments of equivalent changes, which does not affect the essential content of the present invention. Therefore, any simple modifications, equivalent changes and modifications made to the above embodiments based on the technical essence of the present invention without departing from the content of the technical solutions of the present invention are still within the scope of protection of the technical solutions of the present invention.
Claims
1. A tetrapeptide for inhibiting melanin production, characterized in that: Its amino acid sequence is Cys-His-Trp-His.
2. A tetrapeptide for inhibiting melanin production according to claim 1, characterized in that: The confirmation process of the tetrapeptide that inhibits melanin production is as follows: first, a model is constructed by homology modeling, and then screening is performed by bioinformatics methods to analyze the action mode and binding site of the active tetrapeptide with the receptor protein, and the peptide with the best activity is obtained as the tetrapeptide that inhibits melanin production.
3. A tetrapeptide for inhibiting melanin production according to claim 2, characterized in that: The bioinformatics method is molecular docking; The analysis of the action mode and binding site of the active tetrapeptide and the receptor protein was performed using PYMOL software.
4. A method for preparing a tetrapeptide for inhibiting melanin production as claimed in claims 1 to 3, characterized in that: The tetrapeptide for inhibiting melanin production is prepared by solid phase synthesis.
5. The method according to claim 4, characterized in that The following steps are involved: S1, activate WANG resin with three volumes of DCM; S2. Accurately weigh the required His, fully activate it in an ice bath, connect His to WANG resin using the DMAP / DIC / HOBT system, and detect the connection status using the ninhydrin method; S3, washing resin: washing the resin three times with three volumes of DMF / DCM / MEOH; S4, capping the WANG resin with an acetic anhydride:pyridine system, and washing the resin three times with three volumes of DMF / DCM / MEOH; S5, using 20% piperidine solution for deprotection, the deprotection operation is performed twice, and the resin is washed three times with three volumes of DMF / DCM / MEOH, wherein the solvent of the piperidine solution is DMF; S6, repeat the steps of S2, S3, and S5 to connect Trp, His, and Cys in sequence; S7. Cut the resin using a TFA / thioanisole / phenol / water system, cut the connected peptide chains and freeze-dry them to obtain the tetrapeptide that inhibits melanin production.
6. The method according to claim 5, characterized in that The purity of the obtained tetrapeptide for inhibiting melanin production reaches more than 98%.
7. A composition having the effect of inhibiting melanin production, characterized in that: Includes tetrapeptide, niacinamide and alpha-arbutin to inhibit melanin production.
8. The composition having the effect of inhibiting melanin production according to claim 7, characterized in that: Also included is deionized water; The mass proportions of the tetrapeptide, nicotinamide and α-arbutin for inhibiting melanin production in the composition are 0.03-0.05wt%, 0.3-0.5wt% and 0.1-0.3wt% respectively.
9. A method for preparing a composition having the effect of inhibiting melanin production as claimed in claims 7 to 8, characterized in that: The tetrapeptide, nicotinamide and α-arbutin for inhibiting melanin production are mixed in corresponding proportions, and deionized water is added and stirred intermittently to obtain the composition having the effect of inhibiting melanin production. The stirring speed of the intermittent stirring is 200-400 rpm, and the stirring time is 15-20 min.
10. Use of the tetrapeptide for inhibiting melanin production as claimed in claims 1 to 3 and the composition for inhibiting melanin production as claimed in claims 7 to 8 in the preparation of cosmetics.