Composition with skin improving effect, application and cosmetic

Abstract

The invention belongs to the field of new daily chemical materials, and discloses a composition with a skin improving effect, application and cosmetics, the composition comprises acetylhydroxyproline and a white water lily extract, and the invention further discloses a whitening composition. The composition comprises ergothioneine, superoxide dismutase, acetyl hydroxyproline, a white water lily extract and a malt extract. The whitening composition realizes the whitening performance through double saccharification resistance, double oxidation resistance and inhibition of melanin and tyrosinase pathways, and also has potential anti-wrinkle and firming effects at the same time.

Description

Technical Field

[0001] This invention relates to the field of biology, and more particularly to a composition, application, and cosmetic having skin-improving effects. Background Technology

[0002] Patent application CN110585048A, entitled "A composition with anti-glycation and anti-oxidation properties and its application," discloses the use of licorice root extract, nicotinamide, Bright Oléoactif® organic brightening extract, hydrolyzed conchiolin, and plant extracts to enhance the anti-glycation properties of cells.

[0003] The effects of advanced glycation end products (AGEs) on the skin are mainly manifested in their ability to cause skin aging and dullness. There is a direct correlation between anti-glycation and the anti-wrinkle, firming, and whitening properties of products. In ongoing research on several cosmetic raw materials, the applicant has screened out some new raw materials that exhibit excellent anti-glycation effects. Based on this, this application is filed. Summary of the Invention

[0004] The purpose of this invention is to provide a composition that improves skin function. This composition uses acetylhydroxyproline and white water lily extract as the main active ingredients and has a good anti-glycation effect. Meanwhile, the present invention also discloses a whitening composition, which has dual antioxidant properties and also has potential anti-wrinkle and firming effects.

[0005] The present invention also discloses the application of the composition and cosmetics.

[0006] To achieve the above objectives, this application discloses: A composition having skin-improving effects includes acetylhydroxyproline and white water lily extract, wherein the weight ratio of acetylhydroxyproline to white water lily extract is 0.01~2:0.1~5.

[0007] In this invention, acetylhydroxyproline and white water lily extract constitute a dual anti-glycation combination. Acetylhydroxyproline inhibits glycation reactions at the source, reduces the formation of AGEs, and precisely targets the initial stage of glycation. White water lily extract activates the skin whitening pathway, accelerates the metabolism and clearance of AGEs, reduces structural collapse, and improves yellowing. The experiments of this invention further verify that the two have a significant synergistic effect in anti-glycation.

[0008] In the above-mentioned composition with skin-improving effects, the weight ratio of acetylhydroxyproline to white water lily extract is 0.01~1:0.1~3.

[0009] Meanwhile, the present invention also discloses a whitening composition, comprising ergothioneine, superoxide dismutase, acetylhydroxyproline, white water lily extract, and malt extract; The weight ratio of ergothioneine, superoxide dismutase, acetylhydroxyproline, white water lily extract, and malt extract is 0.0001~0.5:0.001~1:0.01~2:0.1~5:0.01~5.

[0010] The compositions of the present invention are broadly classified into two categories: whitening components and dual-antibody components; The brightening component is malt extract; Dual Antioxidant Component: The dual antioxidant component of the present invention is composed of ergothioneine, superoxide dismutase, acetylhydroxyproline, and white water lily extract, wherein ergothioneine and superoxide dismutase exist as a dual antioxidant composition; acetylhydroxyproline and white water lily extract exist as a dual anti-glycation composition; More specifically, ergothioneine: effectively removes ROS, stabilizes the cellular microenvironment, and prevents the initiation of oxidation chain reactions; superoxide dismutase: enhances the endogenous antioxidant capacity of cells and strengthens the skin's own defense system. This invention demonstrates that whitening is achieved through a combination of anti-glycation, tyrosinase inhibition, and melanin inhibition.

[0011] In the above-mentioned whitening composition, the weight ratio of ergothioneine, superoxide dismutase, acetylhydroxyproline, white water lily extract, and malt extract is: 0.0001~0.5: 0.001~1: 0.01~1: 0.1~3: 0.01~3.

[0012] The whitening composition described above also includes a mitochondrial energy-improving composition and / or niacinamide; the mitochondrial energy-improving composition is composed of sunflower bud extract, lemon balm extract, epigallocatechin gallate glucoside, edelweiss callus extract, comfrey root extract, and soapwort extract.

[0013] The weight ratio of sunflower bud extract, lemon balm extract, epigallocatechin gallate glucoside, edelweiss callus extract, comfrey root extract, and soapwort extract in the mitochondrial energy-improving composition is 0.0005~1:0.0005~1:0.0005~1:0.001~1:0.001~1:0.001~1.

[0014] Preferably, the weight ratio of nicotinamide to malt extract is 0.05~0.5:0.01~3, and more preferably, the weight ratio of nicotinamide to malt extract is 0.05~0.5:0.1~3.

[0015] The weight ratio of the mitochondrial energy-improving composition to the malt extract is 0.01 to 0.1:1, more preferably, the weight ratio of the mitochondrial energy-improving composition to the malt extract is 0.01 to 0.05:1; the weight ratio of the mitochondrial energy-improving composition to the malt extract is 0.025:1.

[0016] Furthermore, the present invention also discloses the use of the skin-improving composition as described above, or the use of the whitening composition described above in the preparation of cosmetics.

[0017] Finally, the present invention discloses a cosmetic containing a skin-improving composition as described above, or a whitening composition as described in any of the above.

[0018] In the aforementioned cosmetics, the concentration of the whitening composition is 0.0001~20wt%.

[0019] In the aforementioned cosmetics, the concentration of the whitening composition is 0.001~10wt%; more preferably, the concentration of the whitening composition is 1~5wt%.

[0020] This application has at least the following beneficial effects: 1. The formulation of this invention has been verified to have good anti-glycation and whitening effects through anti-glycation experiments, tyrosinase inhibition experiments, and B16 cell melanin inhibition experiments; at the same time, this invention also has potential anti-wrinkle and firming effects. 2. Through experiments, this invention demonstrates that the invented dual-antibiotic composition can achieve synergistic effects in the presence of malt extract, mitochondrial energy-improving composition, and nicotinamide. Detailed Implementation

[0021] The present invention will now be clearly and completely described in conjunction with embodiments thereof. It should be noted that, unless otherwise specified in the embodiments, conditions are performed under conventional conditions or conditions recommended by the manufacturer. Reagents or instruments whose manufacturers are not specified are all commercially available products. Unless otherwise specified, all parts used in the embodiments of the present invention are parts by weight.

[0022] I. Explanation of Raw Material Sources Ergothioneine powder: Supplier: Zhejiang Huarui Biotechnology Co., Ltd.; Active ingredient content: 95~100wt% Superoxide dismutase powder: Supplier: Shenzhen Zhongke Xinyang Biotechnology Co., Ltd.; Active ingredient content: 0.5~5wt%; Acetylhydroxyproline powder: Supplier: Shenzhen Ruidelin Biotechnology Co., Ltd.; Active ingredient content: 95~100wt%; White water lily extract solution: Supplier: Draco Natural Products, Inc.; Active ingredient content: 10~30 wt%; Malt extract solution: Supplier: RAHN AG; Active ingredient content: 1~10wt%

[0023] Nicotinamide powder: Jiangxi Haiwen Biotechnology Group Co., Ltd.; Active ingredient content: 95-100 wt%

[0024] Actual purchases may involve different manufacturers, and the content may vary slightly.

[0025] Mitochondrial energy improvement composition: prepared by the applicant, traded under the name PCTs, with specific preparation process as described in CN119818418B.

[0026] II. Examples and Comparative Examples

[0027] The preparation methods for each case are as follows: Ergothioneine powder, superoxide dismutase powder, acetylhydroxyproline powder, white water lily extract solution, malt extract solution, PCTs mixture, and nicotinamide powder are dissolved in water and stirred until completely dissolved.

[0028] The components of each case can be found in Table 1 below; Table 1 Formulation Table (Unit: wt%) Ergothioneine powder Superoxide dismutase powder Acetylhydroxyproline powder White water lily extract solution Malt extract solution PCTs mixture Nicotinamide powder Example 1 0 0 0.01 3 0 0 0 Example 2 0 0 0.1 1.5 0 0 0 Example 3 0 0 1 0.1 0 0 0 Comparative Example 1 0 0 0.1 0 0 0 0 Comparative Example 2 0 0 0 1.5 0 0 0 Example 4 0.0001 1 0.01 3 0.1 0 0 Example 5 0.01 0.02 0.2 1.5 1 0 0 Example 6 0.5 0.001 1 0.1 3 0 0 Example 7 0.01 0.02 0.2 1.5 1 0.025 0 Comparative Example 3 0.01 0.02 0 0 1 0 0 Comparative Example 4 0.01 0.02 0.2 0 1 0 0 Comparative Example 5 0.01 0.02 0 1.5 1 0 0 Example 8 0.01 0.02 0.05 0.3 1 0 0.2 Example 9 0.01 0.02 0.05 0.3 1 0.025 0.2 Comparative Example 6 0.01 0.02 0 0.3 1 0 0.2 Comparative Example 7 0.01 0.02 0.05 0 1 0 0.2 Comparative Example 8 0.01 0.02 0 0 1 0 0.2

[0029] III. Performance Testing The testing methods involved in this invention include: anti-glycation test, tyrosinase inhibition rate test, B16 cell melanin inhibition test, and safety test; The specific test methods are as follows: Anti-glycation test: (1) Prepare a mixed solution containing 80 mg / mL bovine serum albumin and 240 mg / mL glucose using PBS, filter it through a 0.22 μm filter membrane, and use it as a 2× saccharification reaction solution.

[0030] (2) Prepare each test sample according to the concentration in the formula table in Table 1, using PBS as solvent, so that the final concentration of the test sample after being added to the reaction system is the concentration in Table 1.

[0031] (3) Prepare each reaction solution in centrifuge tubes according to the reaction system shown in Table 2 below. After mixing evenly, incubate at 55°C for 4 days.

[0032] Table 2 Anti-glycation test reaction system Reaction components (mL) A B C D 2× Glycosylation Reaction Solution 1 0 1 0 2× Sample / Reference Solution 1 1 0 0 PBS 0 1 1 2

[0033] (4) After the reaction is complete, cool the incubated solution to room temperature, centrifuge at 2000 r / min for 5 min, take the supernatant and filter it through a 0.22 μm filter membrane, then take 200 μL of the reaction solution and add it to a 96-well plate. Use a fluorescence microplate reader to detect the fluorescence intensity of each group under the conditions of excitation wavelength 320 nm and emission wavelength 460 nm.

[0034] (5) AGES inhibition rate = [1 - (AB) / (CD)] × 100%.

[0035] Tyrosinase inhibition test: (1) Prepare a tyrosinase solution with a final concentration of 2KU and an L-tyrosine solution with 0.2957mg / mL using PBS.

[0036] (2) Prepare each test sample according to the concentration in the formula table in Table 1, using PBS as solvent, so that the final concentration of the test sample after being added to the reaction system is the concentration in Table 1.

[0037] (3) Set up T (sample group), T0 (sample control group), C (solvent group), and CO (solvent control group). The reaction systems of each group are shown in Table 3 below.

[0038] Table 3. Tyrosinase Inhibition Test Reaction System Reaction components T T0 C C0 Tyrosinase solution (mL) 0.1 / 0.1 / PBS (mL) / 0.1+0.4 2.5 0.1+2.5+0.4 Sample to be tested (mL) 2.5 2.5 / / L-tyrosine solution (mL) 0.4 / 0.4 /

[0039] (4) According to each group of reaction systems, add the corresponding volume of tyrosinase solution, PBS and the sample to be tested to the centrifuge tube respectively. The volume of PBS added to the T0 and C0 groups is 0.1 mL. Mix the solution in the tube thoroughly and incubate at room temperature for 10 min.

[0040] (5) Add 0.4 mL of L-tyrosine solution to the centrifuge tubes of groups T and C respectively, and add 0.4 mL of PBS to the centrifuge tubes of groups T0 and C0 respectively. Mix the solutions in the tubes thoroughly and incubate at room temperature for 10 min.

[0041] (6) After incubation, transfer each group of solutions to a 96-well plate (n=3) and use a multi-functional microplate reader to detect the absorbance value at a wavelength of 470nm.

[0042] (7) Tyrosinase activity inhibition rate = [1-(T-T0) / (C-C0)]×100%.

[0043] B16 cell melanin inhibition test: (1) Inoculate B16 cells into 6-well plates, 1.2 × 10^5 cells per well, and incubate for 24 h.

[0044] (2) Starve the cells for 6 hours using DMEM medium containing 1% FBS.

[0045] (3) Set up a blank control group, a model group, and a sample group. The blank control group was added with DMEM complete medium, the model group was added with DMEM complete medium containing 1 μg / mL MSH, and the sample group was added with DMEM complete medium containing 1 μg / mL α-MSH and the corresponding concentrations of the samples in Table 1. Incubate in an incubator for 60 h.

[0046] (4) The cells in each group were digested with trypsin and collected. The supernatant was removed by centrifugation, and the precipitate was collected and dried. The total protein concentration in each cell precipitate was quantitatively detected by the BCA method, and the total protein content of each group of precipitates was calculated.

[0047] (5) Calculate the total protein content of each group and add an appropriate amount of NaOH solution to make the protein concentration after the precipitation is dissolved equal in each group.

[0048] (6) Heat a metal bath at 70 degrees Celsius for 10 minutes to dissolve and disperse the precipitate into a homogeneous solution. Transfer the solution to a 96-well plate (n=3) and use a multi-functional microplate reader to detect the absorbance at a wavelength of 405 nm.

[0049] (7) Melanin production rate of each group = (OD 实验组 -OD NaOH ) / (OD Control -OD NaOH ) × 100%.

[0050] (8) Melanin improvement rate = (Melanin production rate of model group - melanin production rate of sample group) / (Melanin production rate of model group - melanin production rate of blank group) × 100%.

[0051] Safety Testing: Qualified patch test materials were selected. The test substance (Examples 6 and 7) was placed in the patch tester, with a volume of approximately 0.020 mL to 0.025 mL (the liquid was dropped onto the filter paper attached to the patch tester and placed inside). The patch tester containing the test substance was applied to the back or flexor side of the forearm of the subject using non-irritating adhesive tape. Gently press with the palm of the hand to ensure even application to the skin, and continue for 24 hours. The judgment criteria are shown in Table 4 (Skin Reaction Evaluation Criteria for Closed Patch Tests).

[0052] Table 4. Criteria for evaluating skin reaction in closed patch tests degree of reaction Rating levels Skin reaction - 0 negative reaction ± 1 Suspicious reaction: only slight erythema + 2 Weak positive reaction (erythema reaction): erythema, infiltration, edema, and papules may be present. ++ 3 Strong positive reaction (herpes reaction): erythema, infiltration, edema, papules, vesicles; the reaction may extend beyond the test area. +++ 4 Extremely strong positive reaction (confluent herpes simplex reaction): obvious erythema, severe infiltration, edema, confluent herpes simplex; reaction extends beyond the test area.

[0053] Result determination: If more than 5 out of 30 subjects have a grade 1 adverse skin reaction, or more than 2 out of 30 subjects have a grade 2 adverse skin reaction (more than 5 out of 30 subjects have a grade 2 adverse skin reaction in the patch test of deodorant products), or if any one subject has a grade 3 or higher adverse skin reaction, the test substance is determined to have an adverse skin reaction on the human body. The test results are shown in Tables 5 and 6; Table 5 Test Results AGES inhibition rate % Tyrosinase inhibition rate % Melanin improvement rate % Example 1 26.47 / / Example 2 60.32 / / Example 3 33.62 / / Comparative Example 1 18.00 / / Comparative Example 2 25.44 / / Example 4 / 78.56 / Example 5 / 85.51 / Example 6 / 97.13 / Example 7 / 83.95 / Comparative Example 3 / 63.35 / Comparative Example 4 / 63.35 / Comparative Example 5 / 75.33 / Example 8 / / 77.84 Example 9 / / 72.89 Comparative Example 6 / / 59.57 Comparative Example 7 / / 65.31 Comparative Example 8 / / 54.81

[0054] The security test results are shown in Table 6: Table 6 Results of skin reaction test using closed patch

[0055] Conclusion: The results of the human patch test show that: In Example 30, the skin of the subjects showed a negative reaction (score level 0). According to the judgment criteria of the human patch test in "HAT-SOP-F1-PZ-092", Examples 6 and 7 showed no adverse reactions on human skin.

[0056] Results analysis: As can be seen from Examples 1 to 3 and Comparative Examples 1 and 2 of the present invention, when used alone at a specific concentration, both exhibit a slight AGES inhibition effect. When used in combination, as in Example 2, they exhibit a more obvious AGES scavenging effect. This indicates that the acetylhydroxyproline and white water lily extract of the present invention not only exist as anti-glycation components, but also exhibit a significant synergistic effect. When these two anti-glycation ingredients were applied to a comprehensive formulation, we found equally significant synergistic whitening and melanin-improving effects. Specifically, referring to Examples 4 to 7 and Comparative Examples 3 to 5, in the dual anti-glycation + whitening system, the use of acetylhydroxyproline alone did not improve the whitening effect, but when the two were used in combination, the whitening effect was significantly improved. Similarly, the same conclusion was reached in Examples 8 and 9 and Comparative Examples 6 and 7.

[0057] Based on the above analysis, it can be seen that the dual anti-glycation composition of the present invention has anti-glycation synergy, and when applied to the dual anti-glycation + whitening system, it further exhibits whitening synergy.

Claims

1. A composition having skin-improving effects, characterized in that, It includes acetylhydroxyproline and white water lily extract, wherein the weight ratio of acetylhydroxyproline and white water lily extract is 0.01~2:0.1~5.

2. The composition with skin-improving effects according to claim 1, characterized in that, The weight ratio of acetylhydroxyproline to white water lily extract is 0.01~1:0.1~3.

3. A whitening composition, characterized in that, Including ergothioneine, superoxide dismutase, acetylhydroxyproline, white water lily extract, and malt extract; The weight ratio of ergothioneine, superoxide dismutase, acetylhydroxyproline, white water lily extract, and malt extract is 0.0001~0.5:0.001~1:0.01~2:0.1~5:0.01~5.

4. The whitening composition according to claim 3, characterized in that, The weight ratio of ergothioneine, superoxide dismutase, acetylhydroxyproline, white water lily extract, and malt extract is 0.0001~0.5:0.001~1:0.01~1:0.1~3:0.01~3.

5. The whitening composition according to claim 3, characterized in that, It also includes a mitochondrial energy-enhancing composition and / or nicotinamide; said mitochondrial energy-enhancing composition is composed of sunflower bud extract, lemon balm extract, epigallocatechin gallate glucoside, edelweiss callus extract, comfrey root extract, and soapwort extract; The weight ratio of sunflower bud extract, lemon balm extract, epigallocatechin gallate glucoside, edelweiss callus extract, comfrey root extract, and soapwort extract in the mitochondrial energy-improving composition is 0.0005~1:0.0005~1:0.0005~1:0.001~1:0.001~1:0.001~1.

6. The whitening composition according to claim 5, characterized in that, The weight ratio of nicotinamide to malt extract is 0.05~0.5:0.01~3; the weight ratio of mitochondrial energy-improving composition to malt extract is 0.01~0.1:

1.

7. Use of a composition having skin-improving effects as described in claim 1 or 2, or of a whitening composition as described in any one of 3 to 6, to prepare a cosmetic.

8. A cosmetic product, characterized in that, Contains a skin-improving composition as described in 1 or 2, or a whitening composition as described in any one of 3 to 6.

9. The cosmetic product according to claim 8, characterized in that, The concentration of the whitening composition is 0.0001~20wt%.

10. The cosmetic product according to claim 9, characterized in that, The concentration of the whitening composition is 0.001~10wt%.

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