Anti-wrinkle composition, essence and preparation method of essence

Abstract

The invention discloses an anti-wrinkle composition, essence and a preparation method of the essence, and particularly belongs to the technical field of daily cosmetics. The anti-wrinkle composition is prepared from six core active ingredients, wherein the core active ingredients comprise hydroxyl pinacolone 9-cis retinoate, acetyl hexapeptide-8, palmitoyl tripeptide-5, arginine / lysine polypeptide, saccharomycetes polypeptide and dipeptide diaminobutyrylbenzylamide diacetate, and the anti-wrinkle composition further comprises auxiliary materials acceptable by cosmetics and deionized water. The composition can be prepared into eye essence, cell experiments show that the composition can significantly promote synthesis of type I collagen, human body experiments prove that crow's feet can be reduced and skin elasticity can be increased after the composition is continuously used for 4 weeks, and the composition is suitable for efficient anti-wrinkle nursing of eye crow's feet.

Description

Technical Field

[0001] This invention belongs to the technical field of daily cosmetics, specifically relating to an anti-wrinkle composition, an essence, and a method for preparing the essence. Background Technology

[0002] As we age, the skin around the eyes gradually loses collagen and elastic fibers break down. Combined with factors such as ultraviolet radiation, dryness, and frequent contractions of facial muscles, crow's feet wrinkles, which have both dynamic and static characteristics, are easily formed, seriously affecting appearance.

[0003] However, existing anti-wrinkle cosmetics have many shortcomings and are unable to meet the needs of eye crow's feet care: most products rely on only a single active ingredient. For example, traditional retinol can only promote collagen synthesis for static wrinkles, and single peptides can only alleviate dynamic wrinkles. They cannot achieve the dual effect of "relaxing dynamic wrinkles + repairing static wrinkles", and their effect on improving crow's feet is limited. Traditional retinol-based ingredients are highly irritating and can easily cause redness, dryness and other discomfort when applied directly to sensitive skin around the eyes, resulting in poor applicability. Moreover, existing formulas do not have a scientific multi-component ratio. When some formulas lack key active ingredients, the overall anti-wrinkle effect is significantly reduced, and the synergistic effect between ingredients is not utilized to enhance efficacy.

[0004] Therefore, developing an anti-wrinkle composition that is synergistic, gentle, safe, targeted, and highly effective in reducing crow's feet around the eyes has become an urgent need in the current daily cosmetics industry. Summary of the Invention

[0005] In view of the shortcomings of the prior art, the purpose of this invention is to provide an anti-wrinkle composition and its preparation method that is synergistic in its ingredients, mild and safe, highly targeted and effective in reducing crow's feet wrinkles around the eyes.

[0006] To achieve the above objectives, the present invention discloses the following technical solutions: In a first aspect, the present invention provides an anti-wrinkle composition, characterized in that the composition contains hydroxypinazone 9-cis-retinate, acetyl hexapeptide-8, palmitoyl tripeptide-5, arginine / lysine polypeptide, yeast polypeptide and dipeptide diaminobutyryl benzylamide diacetate. The composition also contains cosmetic-acceptable excipients and solvents.

[0007] Preferably, the composition contains the following components in weight percentages: 0.01-0.015% hydroxypinazone 9-cis-retinate, 0.0002-0.0005% acetyl hexapeptide-8, 0.065-0.085% palmitoyl tripeptide-5, 0.00015-0.0002% arginine / lysine polypeptide, 0.008-0.011% yeast polypeptide, and 0.0001-0.0003% dipeptide diaminobutyryl benzylamide diacetate.

[0008] More preferably, the method for preparing the yeast polypeptide includes the following steps: Step 1. Inoculate the yeast strain into the activation medium and culture it to obtain the activated yeast culture solution; Step 2. Inoculate the yeast culture obtained in Step 1 into the fermentation medium at an inoculation rate of 5-10 v / v%, ferment for 48-60 h, after fermentation, centrifuge to collect the cells, discard the supernatant, resuspend the cells in sterile water, and then homogenize and cycle the resulting slurry at 70 MPa. Centrifuge to collect the supernatant containing the bacterial polypeptides. Step 3. Pass the supernatant containing the bacterial peptides through a 10-30 kDa membrane. The resulting filtrate is then retained using a 1-2 kDa membrane to obtain the retentate. The retentate is then concentrated under vacuum and freeze-dried to obtain the yeast peptides.

[0009] More preferably, in step 1, the yeast strain is inoculated into an activation medium and cultured with shaking at 30°C and 180-220 r / min for 16-18 h to obtain a viable count of 10. 7 -10 8 CFU / mL activated bacterial solution.

[0010] More preferably, the activation culture medium consists of 15-25 g / L glucose, 17-22 g / L peptone, 5-10 g / L yeast extract, and sterile water.

[0011] More preferably, in step 2, the activated yeast culture is inoculated into the fermentation medium at an inoculation rate of 5-10 v / v% based on the volume of the fermentation medium. Fermentation is carried out at 30°C, pH 5.5, 150 r / min, and 30-50% dissolved oxygen for 12 h. Then, the pH is lowered to 3.8, and fermentation continues for 36-48 h. After fermentation, the cells are collected by centrifugation, the supernatant is discarded, the cells are resuspended in sterile water, and then homogenized and circulated at 70 MPa 2-4 times to obtain a slurry. The supernatant containing bacterial polypeptides is collected by centrifugation.

[0012] More preferably, the fermentation medium consists of 40-50 g / L ginseng peptide powder, 30-50 g / L glucose, 10-15 g / L peptone, 5-8 g / L yeast extract, 3-5 g / L ammonium sulfate, 1.5-3 g / L potassium dihydrogen phosphate, 0.5-1 g / L magnesium sulfate heptahydrate, and sterile water.

[0013] Preferably, the excipients are at least one of the following: humectants, thickeners, natural oils, synthetic oils, emulsifiers, pH adjusters, and plant extracts. The solvent is deionized water.

[0014] In a second aspect, the present invention provides an essence for reducing crow's feet around the eyes, the essence containing the anti-wrinkle composition described in the first aspect.

[0015] Thirdly, the present invention provides a method for preparing the serum described in the second aspect, the method comprising the following steps: The essence was obtained by mixing hydroxypinazone 9-cis-retinate, acetyl hexapeptide-8, palmitoyl tripeptide-5, arginine / lysine polypeptide, yeast polypeptide, dipeptide diaminobutyryl benzylamide diacetate, excipients and solvents evenly.

[0016] In this invention: Hydroxypinazone 9-cis-retinoate, as a low-irritant retinoic acid derivative, has the core function of activating fibroblast activity. By regulating the expression of collagen synthesis-related genes such as COL1A1 in cells, it promotes the production and deposition of type I collagen. At the same time, it can also improve the metabolism of the skin stratum corneum and enhance the skin barrier function.

[0017] Acetyl hexapeptide-8, also known as "botox-like peptide," is primarily effective in improving dynamic crow's feet wrinkles. It works by inhibiting the release of acetylcholine at the neuromuscular junction, reducing excessive contraction of muscles such as the orbicularis oculi around the eyes, thereby alleviating the formation and deepening of dynamic wrinkles. Long-term use can also help reduce the formation of static wrinkles (because muscle relaxation can reduce the rate of collagen breakage).

[0018] Palmitoyl tripeptide-5 belongs to the category of "signaling peptides". Its core functions are to repair static wrinkles and strengthen skin structure. It can mimic natural signaling peptides such as transforming growth factor TGF-β in the human body, activate the collagen synthesis pathway of fibroblasts, promote the production of type I and type III collagen and elastic fibers, and at the same time reduce the degradation of collagen by matrix metalloproteinases (MMPs) and maintain the stability of the skin collagen network.

[0019] The core function of arginine / lysine peptides is to optimize the skin microenvironment and assist collagen deposition. Composed of alternating arginine and lysine, it can regulate the skin pH to a slightly acidic physiological environment suitable for the skin around the eyes, promote the proliferation and adhesion of fibroblasts, and provide essential amino acids for collagen synthesis, assisting other ingredients in exerting their collagen-promoting effects.

[0020] The core function of yeast peptides is to promote collagen production and enhance skin resilience. Adding ginseng peptide powder to the fermentation culture medium can provide yeast with a high-quality nitrogen source and active precursors, significantly increasing the content of short active peptides with a molecular weight of 1-2 kDa, such as glutathione and small molecule immune peptides, in the fermentation products. These short peptides can penetrate the skin surface and directly activate the energy metabolism of fibroblasts, accelerating collagen synthesis. At the same time, the addition of ginseng peptide powder can also enhance the antioxidant capacity of yeast peptides and reduce the damage of ultraviolet rays such as UVA to the skin.

[0021] Dipeptide diaminobutyryl benzylamide diacetate has a synergistic effect in combating dynamic wrinkles and protecting collagen. On the one hand, it can block the signal transduction pathway of neuromuscular contraction and complement the target of acetyl hexapeptide-8, thus enhancing the improvement effect of dynamic wrinkles. On the other hand, it inhibits the activity of MMPs and reduces collagen degradation.

[0022] The beneficial effects of this invention; The composition provided by this invention addresses both dynamic and static crow's feet wrinkles. In cell experiments, the growth rate of type I collagen reached over 60%. After 4 weeks of human efficacy testing, the crow's feet wrinkle score significantly decreased (P < 0.01), and skin elasticity was significantly improved (P < 0.001). It has excellent effects in reducing crow's feet wrinkles, improving skin firmness, and slowing down aging. Attached Figure Description

[0023] To better illustrate the purpose, technical solution, and advantages of the present invention, the present invention will be further described below in conjunction with the accompanying drawings.

[0024] Figure 1 Before and after comparison images of subjects using composition 2 for four weeks in human efficacy testing. Detailed Implementation

[0025] To better illustrate the objectives, technical solutions, and advantages of this invention, the invention will be further described below with reference to specific embodiments. Those skilled in the art should understand that the specific embodiments described herein are merely illustrative of the invention and are not intended to limit the invention.

[0026] Unless otherwise specified, the test methods used in the examples and comparative examples are conventional methods; the materials and reagents used are commercially available unless otherwise specified; and the percentages mentioned in the examples and comparative examples are mass percentages unless otherwise specified.

[0027] In this invention:

[0028] Hydroxypinazone 9-cis-retinate: prepared according to Example 1 in patent CN116947723A; Acetyl hexapeptide-8: Purchased from Guangdong Xuanjia Medical Health Technology Co., Ltd.; Palmitoyl tripeptide-5: Purchased from Guangdong Xuanjia Medical Products Health Technology Co., Ltd.; Arginine / Lysine peptides: purchased from Guangdong Xuanjia Medical Health Technology Co., Ltd. Dipeptide diaminobutyryl benzylamide diacetate: purchased from Guangdong Xuanjia Medical Health Technology Co., Ltd.; Ginseng peptide powder: purchased from Zhejiang Yicun Biotechnology Co., Ltd.; Saccharomyces cerevisiae, accession number CICC 1406, purchased from China Industrial Microbial Culture Collection Center; Activation medium: glucose 20 g / L, peptone 20 g / L, yeast extract 10 g / L, and sterile water; Fermentation medium: ginseng peptide powder 45g / L, glucose 30g / L, peptone 10g / L, yeast extract 5g / L, ammonium sulfate 5g / L, potassium dihydrogen phosphate 3g / L, magnesium sulfate heptahydrate 0.5g / L, and sterile water.

[0029] Preparation of yeast polypeptides① Step 1. Pick a small amount of yeast from the slant and inoculate it into an Erlenmeyer flask containing 50 mL of activation medium. Incubate at 30 °C and 180 rpm for 18 h with shaking to obtain a viable count of 10. 8 CFU / mL activated bacterial solution; Step 2. Based on the volume of the fermentation medium, inoculate the activated bacterial solution into the fermentation medium at an inoculation rate of 10 v / v%. Ferment for 12 h at 30℃, pH 5.5, 150 r / min, and 40% dissolved oxygen (DO). Then lower the pH to 3.8 and continue fermentation for 48 h. After fermentation, collect the bacterial cells by centrifugation at 8000 r / min for 10 min, discard the supernatant, resuspend the bacterial cells in sterile water at a mass-to-volume ratio of 1:10 g / mL, and homogenize and circulate the mixture three times under high pressure at 70 MPa. Centrifuge the resulting slurry at 10000 r / min for 20 min and collect the supernatant containing bacterial polypeptides. Step 3. Separate the supernatant through an ultrafiltration membrane. First, use a 10kDa membrane to retain large protein molecules and cell debris. Then, use a 1kDa membrane to retain the filtrate to obtain the retentate. Concentrate the retentate under vacuum and then freeze-dry it to obtain yeast polypeptides.

[0030] Preparation of yeast polypeptide ② The preparation process of yeast polypeptide ① is the same as that of yeast polypeptide ①, except that the "ginseng peptide powder" component in the fermentation medium is omitted. The rest is the same as that of yeast polypeptide ①. The yeast polypeptide obtained is denoted as yeast polypeptide ②.

[0031] Preparation of anti-wrinkle composition The raw materials were accurately weighed according to the mass percentage in Table 1, and homogenized to obtain the anti-wrinkle composition. Table 1. Percentage of Raw Material Mass (Unit: %)

[0032] Note: " / " in the table indicates no addition.

[0033] Performance testing Test on the impact of type I collagen synthesis HDF cells were loaded at 5 × 10 4 Cells were seeded per well into 24-well plates and incubated overnight in an incubator (37°C, 5v / v% CO2) using DMEM (high glucose medium) containing 10% FBS (fetal bovine serum). When the cell layer coverage reached 40%-60%, the original medium was discarded, and the cells were divided into groups for drug administration: 1 mL of fresh medium was added to each well of the blank control and negative control groups, while 1 mL of medium containing 2% (w / w) of the test substance (compositions 1-9) was added to each well of the sample groups, with 6 replicates per group. After drug administration, the 24-well plates were placed in an incubator (37°C, 5v / v% CO2) for further culture. After 24 hours of cell culture, the negative control and sample groups received a total dose of 20 J / cm². 2 The control group was placed under UVA radiation, while the blank control group was placed in the same environment (but with a UVA radiation dose of 0 J / cm²). 2 After irradiation, the cells were cultured in an incubator (37℃, 5v / v% CO2) for 24 hours. The cell culture supernatant was collected, and the collagen I content was analyzed according to the instructions of the ELISA assay kit.

[0034] The growth rate of COL-I content relative to the negative control group was calculated using the following formula, and the results are shown in Table 2.

[0035] Growth rate (%) = [(A1-A0) / A0] × 100% In the formula: A1—Average COL-I content of each sample group, ng·mL -1 ; A0 – Average COL-I content in the negative control group, ng·mL -1 .

[0036] Table 2. Type I collagen content in each group

[0037] Note: "*" indicates a significant difference compared to composition 2, P<0.05, "**" P<0.01, "***" P<0.001.

[0038] Results analysis: As shown in Table 2, the collagen I content level in the negative control group was significantly lower than that in the blank control group, indicating that UVA stimulation was effective.

[0039] Compared to the negative control group, compositions 1-3 significantly increased the content level of collagen I, demonstrating that the complete formulation of the compositions provided by this invention has excellent anti-aging effects.

[0040] Compared to the negative control group, although compositions 4-7 also promoted collagen I synthesis, the effect was significantly different from that of composition 2. This indicates that each component in the compositions provided by the present invention plays an indispensable key role in firming, anti-wrinkle and anti-aging. The control example (compositions 4-7) obtained after deleting hydroxypinazone 9-cis-retinate, palmitoyl tripeptide-5, arginine / lysine polypeptide and yeast polypeptide respectively showed significantly lower collagen synthesis promotion performance than composition 2.

[0041] Compositions 8 and 9 respectively replaced yeast polypeptide ① with yeast polypeptide ② and ginseng peptide powder. The results showed that compared with composition 2, the anti-aging performance of both compositions was significantly reduced, indicating that adding ginseng peptide powder during the preparation of yeast polypeptide can significantly increase the content of active short peptides in yeast polypeptide and more effectively promote collagen synthesis.

[0042] Human efficacy test 1. Subject criteria Forty female participants aged 40-45 years were randomly divided into four groups, each receiving a corresponding test substance. Before the trial, participants were informed of the research objectives, potential benefits, risks, issues, and related rights and obligations. They read and signed informed consent forms. Specific inclusion and exclusion criteria were as follows: 1) Inclusion criteria: female aged 40-45 years, in good health, with no history of photosensitivity diseases, no recent use of drugs that affect photosensitivity, able to understand the trial process, willing to participate in the trial and sign a written informed consent form.

[0043] 2) Exclusion criteria: Women who are pregnant, breastfeeding, or planning to become pregnant during the trial; those with skin diseases at the measurement site that may affect the interpretation of the test results; those who cannot avoid sun exposure when working outdoors; those who have participated in other clinical trials within the past 2 months; those with allergies or high sensitivity to cosmetics; those who have used antihistamines within the past week or immunosuppressants within the past month; those who have applied any anti-inflammatory drugs to the test site within the past two months; those with clinically unhealed inflammatory skin diseases, or those currently receiving treatment for asthma or other chronic respiratory diseases; those who have received chemotherapy for cancer within the past 6 months; those with immunodeficiency or autoimmune diseases, or insulin-dependent diabetes mellitus, or those with highly allergic constitutions; those who have used oral or topical cosmetic products that may affect the test results within the past month; those who are not participating voluntarily or are unable to complete the prescribed content as required by the trial.

[0044] 2. Test substance and testing items 2.1 Test substance The test substances were the aforementioned prepared composition 2, and compositions 2-1, 2-2, and 2-3 obtained by adjusting the formulation of composition 2. Composition 2-1 was prepared by removing acetyl hexapeptide-8 and dipeptide diaminobutyryl benzylamide diacetate from the formulation of composition 2. Composition 2-2 was prepared by removing hydroxypinazone 9-cis-retinoate, palmitoyl tripeptide-5, arginine / lysine polypeptide, and yeast polypeptide ① from the formulation of composition 2. Composition 2-3 was prepared by removing hydroxypinazone 9-cis-retinoate, acetyl hexapeptide-8, palmitoyl tripeptide-5, arginine / lysine polypeptide, dipeptide diaminobutyryl benzylamide diacetate, and yeast polypeptide ① from the formulation of composition 2. The missing amounts were supplemented with solvent.

[0045] 2.2 Projects and Methods Before the test, the subjects cleaned their faces with water and sat quietly for 30 minutes in a laboratory at a temperature of 21±1℃ and 50±10%RH to allow their blood circulation to reach a normal and stable level before proceeding with the subsequent tests.

[0046] Forty subjects were randomly divided into four groups, each receiving one of the test products (compositions 2, 2-1, 2-2, and 2-3). The products were applied twice daily, morning and evening after cleansing, with 2g used each time, for four consecutive weeks. The test products were applied evenly to the eye area and face, gently pressing until absorbed. Other anti-wrinkle skincare products were discontinued during the trial. The following tests were performed on the subjects before and after four weeks of use to compare differences before and after the initial application.

[0047] Crow's feet wrinkle rating: Professional examiners used the Asian Skin Aging Atlas Vol.02 (Chinese version) to rate the crow's feet wrinkles around the eyes of the subjects. Each indicator was rated from 0 (very good) to 5 (very poor) according to its severity.

[0048] Skin elasticity measurement: The R2 value of the skin elasticity around the eyes of the subjects was measured using a Cutometer dual MPA580. The measurement was performed 3 times and the average value was taken. The closer the R2 value is to 1, the better the elasticity.

[0049] 3. Data Analysis Data were analyzed for differences and correlations using SPSS 21.0.

[0050] 4. Experimental Results Table 3

[0051] Note: "*" indicates within-group comparison, with significant difference (P < 0.05), "**" P < 0.01, "***" P < 0.001.

[0052] 5 Results Analysis Within-group comparisons showed that after 4 weeks of use, the crow's feet score of composition 2 decreased significantly (P < 0.01), and the crow's feet of the subjects improved significantly (see [link to article]). Figure 1 The results show that the complete formula can effectively reduce the degree of crow's feet wrinkles; the skin elasticity R2 value of composition 2 increased from the initial 0.48 to 0.68, which significantly improved the skin elasticity (P<0.001), which proves that the complete formula can significantly repair the elastic fiber network of the skin around the eyes and enhance skin firmness.

[0053] Finally, it should be noted that the above descriptions are merely preferred embodiments of the present invention and are not intended to limit the present invention. Although the present invention has been described in detail with reference to the foregoing embodiments, those skilled in the art can still modify the technical solutions described in the foregoing embodiments or make equivalent substitutions for some of the technical features. Any modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of the present invention should be included within the protection scope of the present invention.

Claims

1. An anti-wrinkle composition, characterized in that, The composition contains the following components in weight percentages: 0.01-0.015% hydroxypinazone 9-cis retinate, 0.0002-0.0005% acetyl hexapeptide-8, 0.065-0.085% palmitoyl tripeptide-5, 0.00015-0.0002% arginine / lysine polypeptide, 0.008-0.011% yeast polypeptide, and 0.0001-0.0003% dipeptide diaminobutyryl benzylamide diacetate; The composition also contains cosmetic-acceptable excipients and solvents; The preparation method of the yeast polypeptide includes the following steps: Step 1. Inoculate the yeast strain into the activation medium and culture it to obtain the activated yeast culture solution; Step 2. Inoculate the yeast culture obtained in Step 1 into the fermentation medium at an inoculation rate of 5-10 v / v%, ferment for 48-60 h, after fermentation, centrifuge to collect the cells, discard the supernatant, resuspend the cells in sterile water, and then homogenize and cycle the resulting slurry at 70 MPa. Centrifuge to collect the supernatant containing the bacterial polypeptides. Step 3. Pass the supernatant containing the bacterial peptides through a 10-30 kDa membrane. The resulting filtrate is then retained using a 1-2 kDa membrane to obtain the retentate. The retentate is then concentrated under vacuum and freeze-dried to obtain the yeast peptides.

2. The anti-wrinkle composition according to claim 1, characterized in that, In step 1, the yeast strain is inoculated into the activation medium and cultured at 30℃ and 180-220 r / min for 16-18 h with shaking, resulting in a viable count of 10. 7 -10 8 CFU / mL activated bacterial solution.

3. The anti-wrinkle composition according to claim 2, characterized in that, The activation culture medium consists of 15-25 g / L glucose, 17-22 g / L peptone, 5-10 g / L yeast extract, and sterile water.

4. The anti-wrinkle composition according to claim 1, characterized in that, In step 2, based on the volume of the fermentation medium, the activated yeast culture is inoculated into the fermentation medium at an inoculation rate of 5-10 v / v%. Fermentation is carried out at 30℃, pH 5.5, 150 r / min, and 30-50% dissolved oxygen for 12 h. Then, the pH is lowered to 3.8, and fermentation continues for 36-48 h. After fermentation, the cells are collected by centrifugation, the supernatant is discarded, the cells are resuspended in sterile water, and then homogenized and circulated at 70 MPa 2-4 times to obtain a slurry. The supernatant containing bacterial polypeptides is collected by centrifugation.

5. The anti-wrinkle composition according to claim 4, characterized in that, The fermentation medium consists of 40-50 g / L ginseng peptide powder, 30-50 g / L glucose, 10-15 g / L peptone, 5-8 g / L yeast extract, 3-5 g / L ammonium sulfate, 1.5-3 g / L potassium dihydrogen phosphate, 0.5-1 g / L magnesium sulfate heptahydrate, and sterile water.

6. The anti-wrinkle composition according to claim 1, characterized in that, The excipients are at least one of the following: humectants, thickeners, natural oils, synthetic oils, emulsifiers, pH adjusters, and plant extracts. The solvent is deionized water.

7. A serum for reducing crow's feet around the eyes, characterized in that, The serum contains the anti-wrinkle composition according to any one of claims 1-6.

8. A method for preparing the serum according to claim 7, characterized in that, The preparation method includes the following steps: The essence was obtained by mixing hydroxypinazone 9-cis-retinate, acetyl hexapeptide-8, palmitoyl tripeptide-5, arginine / lysine polypeptide, yeast polypeptide, dipeptide diaminobutyryl benzylamide diacetate, excipients and solvents evenly.

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