Composition containing anoectochilus formosanus extract and tetrapeptide and having anti-wrinkle and firming effects and application thereof
By combining the nigra extract with a variety of peptide complexes, a composition with anti-wrinkle firming effect was developed, which solved the problem of limited effects of existing cosmetics in wrinkles and skin firming, and achieved significant results synergistic and human verification is good.
Patent Information
- Application Number
- CN202411566736.5
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2024-11-05
- Publication Date
- 2025-02-07
AI Technical Summary
Existing cosmetics are difficult to effectively solve the problems of wrinkles and skin firming, and most ingredients have limited effects when used alone, and lack the combined use of synergies.
Develop a composition consisting of a genusa extract and a tetrapeptide (including palmitoyl tripeptide-5, decarboxylic carnosine hydrochloride, arginine/lysine polypeptide and acetyl hexapeptide-8) by The matching technology improves the anti-wrinkle firming effect of cosmetics.
This composition significantly improves the anti-wrinkle firming effect of cosmetics through synergistic effects, meets the market's demand for efficient and multifunctional skin care products, and shows good anti-wrinkle effects in human verification.
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Figure CN119385905A_ABST
Abstract
Description
Technical Field
[0001] The present invention relates to the technical field of cosmetic formulas, and in particular to a composition containing a roxburghii roxburghii extract and a tetrapeptide and having anti-wrinkle and firming effects, and application thereof. Background Art
[0002] Anoectochilus roxburghii is a perennial herbaceous plant of the genus Anoectochilus in the Orchidaceae family. Also known as golden thread orchid, it also has other names such as bird ginseng, golden thread tiger-head banana, and golden thread bone-clearing. It is mentioned in the Chinese Materia Medica Encyclopedia, the National Compendium of Chinese Herbal Medicines, the Chinese Materia Medica, and the Fujian Materia Medica. It is rich in polysaccharides, steroids, organic acids, terpenes, alkaloids, volatile oils, flavonoids, and glycosides, and exhibits anti-inflammatory, anti-tumor, antioxidant, and anti-HBV activities.
[0003] Palmitoyl Tripeptide-5 can smooth wrinkles and tighten the skin by stimulating the production of collagen and elastin in the skin. It also improves skin firmness and hydration, restoring smoothness and elasticity. It is a small molecule peptide widely used in anti-wrinkle and firming cosmetics.
[0004] Decarboxylated carnosine hydrochloride is a chemical ingredient with significant antioxidant properties, protecting collagen and elastin from oxidative stress, thereby maintaining healthy skin structure. Furthermore, it possesses a stronger positive charge and does not form intramolecular salts, resulting in improved adsorption and permeability on the skin surface. Therefore, it is widely used in cosmetics and personal care products.
[0005] Arginine / lysine peptides, also known as conopeptides or conotoxins, are peptides composed of the amino acids arginine and lysine with diverse applications in cosmetics and personal care products. Arginine / lysine peptides are sequenced and synthesized to exhibit highly potent muscle relaxants. This muscle relaxant effect can rapidly reduce the appearance of dynamic wrinkles on the skin's surface, providing an immediate anti-wrinkle effect.
[0006] Acetyl Hexapeptide-8, also known as Argireline or Hexapeptide, is a neurotransmitter-inhibiting biomimetic peptide composed of six amino acids. Its primary mechanism of action is to block the release of neurotransmitters, reducing muscle contraction and achieving anti-wrinkle and anti-aging effects. It also promotes collagen production, enhancing skin elasticity and radiance, and increasing skin moisture and hydration.
[0007] Anoectochilus roxburghii extract, palmitoyl tripeptide-5, decarboxylated carnosine hydrochloride, arginine / lysine polypeptide, and acetyl hexapeptide-8 are safe and mild to all skin types. The present invention first conducts cell experiments on Anoectochilus roxburghii extract, palmitoyl tripeptide-5, decarboxylated carnosine hydrochloride, arginine / lysine polypeptide, and acetyl hexapeptide-8, and finds that they have a synergistic effect. Subsequently, human efficacy verification is conducted, and it is found that they have good anti-wrinkle efficacy. Therefore, the development of a composition of Anoectochilus roxburghii extract, palmitoyl tripeptide-5, decarboxylated carnosine hydrochloride, arginine / lysine polypeptide, and acetyl hexapeptide-8 has great development prospects. Summary of the Invention
[0008] The purpose of the present invention is to provide a composition containing a roxburghii extract and a peptide complex with anti-wrinkle and firming effects and its application.
[0009] To achieve the above object, the present invention adopts the following technical solutions: A composition containing aroximate roxburghii extract and a tetrapeptide with anti-wrinkle and firming effects. The composition is composed of a compound of aroximate roxburghii extract and a tetrapeptide; the tetrapeptide is palmitoyl tripeptide-5, decarboxylated carnosine hydrochloride, arginine / lysine polypeptide, and acetyl hexapeptide-8; the aroximate roxburghii extract, palmitoyl tripeptide-5, decarboxylated carnosine hydrochloride, arginine / lysine polypeptide, and acetyl hexapeptide-8 are compounded in a mass ratio of (750-2500): 500:500:1:5.
[0010] The preparation method of the above-mentioned roxburghii extract comprises the following steps: freeze-drying and crushing the fresh roxburghii herb, extracting it with water 1-2 times at a mass ratio of freeze-dried powder to pure water of 1:10 and a temperature of 10-40°C, with each extraction time being 1-2 hours, and concentrating it under reduced pressure and low temperature (temperature of 45°C-55°C and pressure range of 10 -2 -10 -5 mbar) to a sugar content of 65%-75%, and then filtered through 300-mesh gauze to obtain a roxburghii roxburghii extract, whose main active ingredient is total polysaccharide with a content of 10wt%-20wt%. The above composition is used in anti-wrinkle and firming cosmetics, wherein the concentrations of the roxburghii extract, palmitoyl tripeptide-5, decarboxylated carnosine hydrochloride, arginine / lysine polypeptide and acetyl hexapeptide-8 in the cosmetics are 0.15-0.5wt%, 0.1wt%, 0.1wt%, 0.0002wt% and 0.001wt%.
[0011] The beneficial effects of the present invention are: The combination of Anoectochilus roxburghii extract, palmitoyl tripeptide-5, decarboxylated carnosine hydrochloride, arginine / lysine peptide, and acetyl hexapeptide-8 synergistically enhances the overall efficacy of cosmetics, meeting market demand for highly effective and multifunctional skin care products. BRIEF DESCRIPTION OF THE DRAWINGS
[0012] Figure 1 : Comparison of the anti-wrinkle and firming effects of a combination of roxburghii extract and a peptide complex over 28 days (left: before use; right: after use). DETAILED DESCRIPTION
[0013] To make the above features and advantages of the present invention more clearly understood, the following embodiments are given for detailed description. Unless otherwise specified, the methods of the present invention are all conventional methods in the art.
[0014] The present invention was tested by the following experiments and standards: 1. Based on the human dermal fibroblast (HDF-α) model, the anti-wrinkle and firming efficacy was evaluated by detecting the changes in the levels of tissue inhibitor of metalloproteinases, sirtuin 1, fibronectin, and acetylcholine after the dermal fibroblasts were treated with roxburghii extract, palmitoyl tripeptide-5, decarboxylated carnosine hydrochloride, arginine / lysine peptide, and acetyl hexapeptide-8.
[0015] 2. Use a facial image analyzer to collect two-dimensional images, and analyze the effect of the composition on improving wrinkles around the eyes through the image analysis.
[0016] Example 1 Preparation of Anoectochilus roxburghii Extract The preparation method of the extract of Anoectochilus roxburghii comprises the following steps: freeze-drying and crushing the fresh herb of Anoectochilus roxburghii, extracting it twice with water at a mass ratio of freeze-dried powder to pure water of 1:10 and a temperature of 35°C, with each extraction time of 1.5 hours, and concentrating it under reduced pressure and low temperature (temperature of 50°C and pressure range of 10 -3 mbar) to a sugar content of 70% and then filtered through 300-mesh gauze to obtain a roxburghii roxburghii extract, the main active ingredient of which was total polysaccharide with a content of 10 wt%.
[0017] Example 2 Cell activity test 1. Preparation of mother liquor of each component 1) Preparation of Anoectochilus roxburghii extract solution: Weigh 0.2 g of the Anoectochilus roxburghii extract obtained in Example 1 and add 1.8 mL of DMEM complete medium to prepare a 10% Anoectochilus roxburghii extract stock solution. Dilute with DMEM complete medium to the appropriate concentration before use.
[0018] 2) Preparation of Palmitoyl Tripeptide-5 Solution: Take 0.2 mL of Palmitoyl Tripeptide-5 and add 1.8 mL of DMEM complete medium to prepare a 10% Palmitoyl Tripeptide-5 stock solution. Dilute with DMEM complete medium to the corresponding concentration before use.
[0019] 3) Preparation of decarboxylated carnosine hydrochloride solution: Take 0.2 mL of decarboxylated carnosine hydrochloride and add 1.8 mL of DMEM complete medium to prepare a 10% decarboxylated carnosine hydrochloride stock solution. Dilute with DMEM complete medium to the appropriate concentration before use.
[0020] 4) Preparation of Arginine / Lysine Peptide Solution: Pipette 0.02 mL of Arginine / Lysine Peptide and add 1.98 mL of DMEM complete medium to prepare a 1% Arginine / Lysine Peptide stock solution. Dilute to the appropriate concentration with DMEM complete medium before use.
[0021] 5) Preparation of Acetyl Hexapeptide-8 Solution: Weigh 0.02 g of Acetyl Hexapeptide-8 and add 1.98 mL of DMEM complete medium to prepare a 1% Acetyl Hexapeptide-8 stock solution. Dilute with DMEM complete medium to the appropriate concentration before use.
[0022] 2. Cell activity test (MTT method) 1) Plating: Take cultured human dermal fibroblasts, digest and centrifuge, count, and prepare the cell concentration to 3×10 5 pcs / mL, pipette evenly, spread into 96-well plates, 100 μL / well, and incubate in an incubator for 24 h ± 2 h.
[0023] 2) Administration: After 24 h ± 2 h, add the prepared sample solution (at a concentration twice the test concentration) at 100 μL per well in triplicate. Add the same volume of culture medium (with cells) to the blank group and culture medium only (without cells) to the solvent group. Continue incubation for 24 h ± 2 h.
[0024] 3) Test: After incubation for 24 hours, discard the supernatant and add 200 μL of 0.5 mg / mL MTT solution to each well. Place in a CO2 incubator for 4 hours.
[0025] 4) Test: After 4 hours, the supernatant was discarded and 150 μL of DMSO was added to each well. The cells were shaken in the dark at room temperature for 10 minutes. The absorbance was measured at a wavelength of 490 nm. The MTT test results are shown in Table 1.
[0026] Table 1 Results of MTT assay for cell viability As shown in Table 1, the maximum addition amount of golden thread lotus extract is 0.7%, the maximum addition amount of palmitoyl tripeptide-5 is 0.1%, the maximum addition amount of decarboxylated carnosine hydrochloride is 0.1%, the maximum addition amount of arginine / lysine polypeptide solution is 0.0002%, and the maximum addition amount of acetyl hexapeptide-8 is 0.001%.
[0027] Example 3 Determination of Anti-wrinkle and Firming Efficacy 1. Based on the UVA irradiated human dermal fibroblast (HDF-α) model, enzyme-linked immunosorbent assay was used to detect the changes in the contents of tissue inhibitor of metalloproteinases, sirtuin 1, fibronectin, and acetylcholine to evaluate the anti-wrinkle and firming effects of roxburghii extract, palmitoyl tripeptide-5, decarboxylated carnosine hydrochloride, arginine / lysine polypeptide, acetyl hexapeptide-8, tetrapeptide combination, and the combination of roxburghii extract and tetrapeptide combination.
[0028] Step 1: Single-component anti-wrinkle and firming efficacy test (1) Plating: Digest and centrifuge the cultured cells, count them, and set the cell concentration to 3×10 5 Pipette evenly, add 1 mL to each well of a 12-well plate, and incubate in an incubator for 24 ± 2 h.
[0029] (2) Administration: Administer the drug 24 ± 2 hours later. Return the cells to a 37°C, 5% CO2 incubator and continue incubation for 24 hours.
[0030] (3) Irradiation: 24 hours after administration, UVA irradiation was performed with an irradiation intensity of 125 mJ / cm 2 After irradiation, the cells were given the drug again and then placed back in a 37°C, 5% CO2 incubator for 24 hours.
[0031] (4) Test: After 24 h of culture, the cell supernatant was collected and the contents of tissue inhibitor of metalloproteinases, sirtuin 1, fibronectin, and acetylcholine were tested using an ELISA kit. The experimental steps are as follows: 1) Remove the tissue inhibitor of metalloproteinases, sirtuin 1, fibronectin, and acetylcholine kits from the refrigerator 1 hour in advance and equilibrate to room temperature. Remove the collected supernatant from the -80°C freezer in advance, thaw, and centrifuge at 10,000 rpm for 20 minutes.
[0032] 2) Remove the strips from the aluminum foil pouch and set up standard and sample wells. Add 50 μL of each of the six concentrations of standard to the standard wells, 50 μL of each collected cell supernatant to the sample wells, and 100 μL of HRP enzyme-labeled reagent to both the standard and sample wells. Cover with sealing film and incubate at 37°C for 60 min.
[0033] 3) After 60 minutes, remove the ELISA plate, discard the liquid, add 350 μL of wash buffer to each well, let it stand for 1 minute, discard the wash buffer, pat dry on absorbent paper, and repeat the wash five times. Add 50 μL each of substrates A and B to each well, cover with sealing film, and incubate at 37°C in a constant temperature incubator for 15 minutes.
[0034] 4) After 15 minutes, remove the plate and add 50 μL of stop solution to each well to terminate the color development reaction. Measure the absorbance at 450 nm within 15 minutes.
[0035] 5) Create a curve with the absorbance OD value as the ordinate (Y) and the concentration of the corresponding analyte standard as the abscissa (X). Calculate the analyte concentration in the sample based on its OD value using the standard curve and then calculate the corresponding upregulation / inhibition rate.
[0036] Table 2 Test results of single-component anti-wrinkle and firming efficacy As shown in Table 2, roxburghii extract, palmitoyl tripeptide-5, decarboxylated carnosine hydrochloride, arginine / lysine peptide solution, and acetyl hexapeptide-8 can promote the production of tissue inhibitor of metalloproteinases, sirtuin 1, and fibronectin to a certain extent, inhibit the production of acetylcholine, and have anti-wrinkle and firming effects.
[0037] Step 2: Anti-wrinkle and firming efficacy test of tetrapeptide compositions at different concentrations The following five tetrapeptide compositions with different test concentration ratios were tested for their anti-wrinkle and firming efficacy according to the experimental procedure of step 1.
[0038] 1) Tetrapeptide composition 1 Palmitoyl tripeptide-5: decarboxylated carnosine hydrochloride: arginine / lysine peptide solution: acetyl hexapeptide-8 = 100:100:1:1 (i.e. 0.02%: 0.02%: 0.0002%: 0.0002%); 2) Tetrapeptide composition 2 Palmitoyl tripeptide-5: decarboxylated carnosine hydrochloride: arginine / lysine peptide solution: acetyl hexapeptide-8 = 300: 300: 1: 3 (i.e. 0.06%: 0.06%: 0.0002%: 0.0006%); 3) Tetrapeptide composition 3 Palmitoyl tripeptide-5: decarboxylated carnosine hydrochloride: arginine / lysine peptide solution: acetyl hexapeptide-8 = 500: 500: 1: 5 (i.e. 0.1%: 0.1%: 0.0002%: 0.001%); 4) Tetrapeptide composition 4 Palmitoyl tripeptide-5: decarboxylated carnosine hydrochloride: arginine / lysine peptide solution: acetyl hexapeptide-8 = 600: 600: 1: 7 (i.e. 0.096%: 0.096%: 0.00016%: 0.001%); 5) Tetrapeptide composition 5 Palmitoyl tripeptide-5: decarboxylated carnosine hydrochloride: arginine / lysine peptide solution: acetyl hexapeptide-8 = 700: 700: 1: 5 (i.e. 0.098%: 0.098%: 0.00014%: 0.0007%).
[0039] Table 3 Anti-wrinkle and firming efficacy test results of tetrapeptide composition Combining Tables 2 and 3, it can be seen that the anti-wrinkle and firming efficacy test results of the tetrapeptide combination are superior to those of palmitoyl tripeptide-5, decarboxylated carnosine hydrochloride, arginine / lysine peptide solution, and acetyl hexapeptide-8 alone. The optimal ratio of tetrapeptide combination 3, namely palmitoyl tripeptide-5: decarboxylated carnosine hydrochloride: arginine / lysine peptide solution: acetyl hexapeptide-8, is 0.1%: 0.1%: 0.0002%: 0.001%.
[0040] Step 3: Anti-wrinkle and firming efficacy test of different concentrations of Anoectochilus roxburghii tetrapeptide compositions The following three different test concentration ratios of the roxburghii extract and the tetrapeptide composition were used to perform anti-wrinkle and firming efficacy tests according to the experimental procedure of step 1.
[0041] 1) Anoectochilus roxburghii tetrapeptide composition 1 Golden thread extract: palmitoyl tripeptide-5: decarboxylated carnosine hydrochloride: arginine / lysine peptide solution: acetyl hexapeptide-8 = 750: 500: 500: 1: 5 (i.e. 0.15%: 0.1%: 0.1%: 0.0002%: 0.001%); 2) Anoectochilus roxburghii tetrapeptide composition 2 Anoectochilus roxburghii extract: palmitoyl tripeptide-5: decarboxylated carnosine hydrochloride: arginine / lysine peptide solution: acetyl hexapeptide-8 = 1500: 500: 500: 1: 5 (i.e. 0.3%: 0.1%: 0.1%: 0.0002%: 0.001%); 3) Anoectochilus roxburghii tetrapeptide composition 3 Anoectochilus roxburghii extract: palmitoyl tripeptide-5: decarboxylated carnosine hydrochloride: arginine / lysine peptide solution: acetyl hexapeptide-8 = 2500: 500: 500: 1: 5 (i.e. 0.5%: 0.1%: 0.1%: 0.0002%: 0.001%); Table 4 Test results of the anti-wrinkle and firming efficacy of the Anoectochilus roxburghii tetrapeptide composition Combining Tables 3 and 4, it can be seen that the anti-wrinkle and firming efficacy test results of the combination of roxburghii extract and tetrapeptide are superior to those of the tetrapeptide combination. When the concentration ratio of roxburghii extract, palmitoyl tripeptide-5, decarboxylated carnosine hydrochloride, arginine / lysine peptide, and acetyl hexapeptide-8 is within the range of (750-2500):500:500:1:5, the five active ingredients can produce a synergistic effect and exert significant anti-wrinkle and firming effects. Among them, the ratio of roxburghii extract: palmitoyl tripeptide-5: decarboxylated carnosine hydrochloride: arginine / lysine peptide solution: acetyl hexapeptide-8 = 1500:500:500:1:5 (i.e. 0.3%:0.1%:0.1%:0.0002%:0.001%) has the best effect.
[0042] 2. Determination of the efficacy of improving fine lines around the eyes The determination of improvement of fine lines around the eyes was based on 30 Asian adult subjects (age: 35.64±0.52) using a 0.15% carbomer hydrogel system product containing a roxburghii extract complex (roxburghii extract, palmitoyl tripeptide-5, decarboxylated carnosine hydrochloride, arginine / lysine peptide and acetyl hexapeptide-8 at a concentration ratio of 750:500:500:1:5=0.15%:0.1%:0.1%:0.0002%:0.001%) for 28 consecutive days under normal conditions. The skin wrinkle improvement rate was measured using the PRIMOS-CR instrument to evaluate the anti-wrinkle efficacy of the composition.
[0043] The results are as follows Figure 1 As shown in the results, after 28 days of using a 0.15% carbomer hydrogel system product containing a complex of roxburghii extract (roxburghii extract, palmitoyl tripeptide-5, decarboxylated carnosine hydrochloride, arginine / lysine peptide and acetyl hexapeptide-8 at a concentration ratio of 750:500:500:1:5 = 0.15%:0.1%:0.1%:0.0002%:0.001%), fine lines around the eyes were significantly improved, with an improvement rate of 15.81% ( P <0.05).
[0044] The above description is only a preferred embodiment of the present invention. All equivalent changes and modifications made according to the scope of the patent application of the present invention should fall within the scope of the present invention.
Claims
1. A composition containing aroximate extract and tetrapeptide with anti-wrinkle and firming effects, characterized in that: The composition is prepared by compounding a roxburghii extract and tetrapeptides; the tetrapeptides are palmitoyl tripeptide-5, decarboxylated carnosine hydrochloride, arginine / lysine polypeptide and acetyl hexapeptide-8; the roxburghii extract, palmitoyl tripeptide-5, decarboxylated carnosine hydrochloride, arginine / lysine polypeptide and acetyl hexapeptide-8 are compounded in a mass ratio of (750-2500): 500: 500: 1:
5.
2. The composition according to claim 1, characterized in that The preparation method of the roxburghii extract comprises the following steps: fresh roxburghii grass is freeze-dried and crushed, extracted 1-2 times under the conditions that the mass ratio of freeze-dried powder to pure water is 1:10 and the temperature is 10-40°C, the extraction time for each time is 1-2h, and the extraction is terminated when the sugar content is 65%-75% under reduced pressure and low temperature, and filtered with 300-mesh gauze to obtain the roxburghii extract, wherein the main active component is total polysaccharide with a content of 10wt%-20wt%.
3. The composition according to claim 2, characterized in that: The temperature during low-temperature concentration is 45℃-50℃ and the pressure range is 10 -2 -10 -5 mbar.
4. Use of the composition according to any one of claims 1 to 2 in anti-wrinkle and firming cosmetics, characterized in that: The application concentrations of the roxburghii extract, palmitoyl tripeptide-5, decarboxylated carnosine hydrochloride, arginine / lysine polypeptide and acetyl hexapeptide-8 in cosmetics are 0.15%-0.5%wt, 0.1wt%, 0.1wt%, 0.0002wt% and 0.001wt%.