An anti-aging composition and its application in cosmetics

Abstract

The present invention discloses an anti-aging composition and its application in cosmetics, wherein the composition comprises polyglutamic acid and its salt, hyaluronic acid and its salt, and sweet orange extract. The present invention compounds polyglutamic acid and its salt, hyaluronic acid and its salt, and sweet orange extract to have a synergistic effect, and the overall addition amount thereof can show the effect of improving skin elasticity within a relatively low range, thereby achieving the purpose of reducing wrinkles, tightening skin, and anti-aging.

Description

Technical Field

[0001] The invention relates to a composition, in particular to an anti-aging composition containing polyglutamic acid and its salt, hyaluronic acid and its salt, and sweet orange extract, and also to application of the composition in cosmetics, belonging to the technical field of cosmetics. Background Art

[0002] The loss of skin elasticity is a natural process of human aging. The dermis of human skin contains a huge number of fibroblasts, and the function of fibroblasts is to continuously produce collagen, hyaluronic acid, elastic fibers and other substances that support the youthful state of the skin. The skin of young people looks smooth and elastic because of the presence of a large number of fibroblasts. When people reach a certain age, the fibroblasts in the skin begin to be lost, which correspondingly leads to a decrease in collagen, elastic fibers, hyaluronic acid and other substances, and the skin ages. Therefore, the decline in the activity and number of fibroblasts is one of the fundamental reasons for the loss of skin elasticity and aging. Improving the proliferation ability of fibroblasts can improve skin elasticity and resist aging.

[0003] In the prior art, CN113209271A discloses a composition that can promote the proliferation of fibroblasts and a preparation method thereof. The composition is composed of peptides with the sequences of Gly-Asn-Pro-Asn-Asn-Cly-Ile, Gly-Pro-Hyp-Gly-Ser-Pro, Gly-Leu-Hyp-Gly-Hyp-Ala, Gly-His-Lys-Val-Lys and desmosin and isodesmosin. The ingredients used are not easy to obtain, the preparation method is complicated and the price is expensive.

[0004] The anti-aging products currently on the market generally have complex ingredients and high costs. Some products add irritating active ingredients to enhance the anti-aging effects, which can cause adverse skin irritation reactions. Summary of the invention

[0005] In view of the current status of the prior art, the present invention provides an anti-aging composition, which comprises three effective ingredients. The ingredient types are simple and the ingredients work synergistically to enhance the synergy of the three ingredients, thereby increasing the proliferation of fibroblasts and promoting the synthesis of collagen, thereby having the effects of reducing wrinkles, tightening the skin and improving skin elasticity.

[0006] This application provides the following technical solutions:

[0007] 1. An anti-aging composition, characterized in that the composition comprises polyglutamic acid and its salts, hyaluronic acid and its salts, and sweet orange extract.

[0008] 2. The anti-aging composition according to claim 1, characterized in that the anti-aging composition comprises the following components in parts by weight:

[0009] 0.01-1.0 parts of polyglutamic acid and its salts, 0.01-1.0 parts of hyaluronic acid and its salts, and 0.0015-1.35 parts of sweet orange extract.

[0010] 3. The anti-aging composition according to claim 1 or 2, characterized in that it comprises the following components in parts by weight: 0.01-0.3 parts of polyglutamic acid and its salts, 0.01-0.3 parts of hyaluronic acid and its salts, and 0.015-0.9 parts of sweet orange extract.

[0011] 4. The anti-aging composition according to any one of claims 1 to 3, characterized in that the molecular weight of hyaluronic acid and its salt is 1.5×10 6 Da or less, preferably 1.0×10 6 Da below.

[0012] 5. The anti-aging composition according to any one of claims 1 to 4, characterized in that the molecular weight of polyglutamic acid and its salt is 1.0×10 4 Da-2.0×10 6 Da.

[0013] 6. The anti-aging composition according to any one of claims 1 to 5, characterized in that it also contains auxiliary materials and / or functional ingredients commonly used in the cosmetics field.

[0014] 7. Use of the anti-aging composition according to any one of claims 1 to 6 in the preparation of a product for promoting fibroblast proliferation.

[0015] 8. Use of the anti-aging composition according to any one of claims 1 to 6 in the preparation of cosmetics.

[0016] 9. The use according to claim 8, characterized in that the anti-aging composition is used to prepare anti-aging cosmetics.

[0017] Compared with the prior art, the present invention has the following beneficial effects:

[0018] The present invention combines three components of polyglutamic acid and its salt, hyaluronic acid and its salt, and sweet orange extract, and finds that the components after compounding are synergistic, can enhance the proliferation ability of fibroblasts, has significant antioxidant capacity and the effect of enhancing skin elasticity, and has low irritation, can restore the skin to a smooth and soft feel, restore elastic skin, and greatly reduce skin wrinkles. In addition, the composition is not only superior in efficacy, but also low in price, better in stability, and relatively simple in production and operation process, which greatly improves production efficiency and reduces costs, and has great advantages in both industrial production and consumer markets. BRIEF DESCRIPTION OF THE DRAWINGS

[0019] Figure 1 The zebrafish embryos of the anti-aging composition essence of Example 1 and the blank control group were photographed and shown;

[0020] Figure 2 This is a bar graph of the ROS signal intensity test results of the anti-aging composition essence in Example 1 and the blank control group. DETAILED DESCRIPTION

[0021] In order to better illustrate the technical solutions and advantages of the present invention, the present invention will be further described below in conjunction with specific embodiments. It should be noted that the process parameters and process steps not described in detail in the present invention are all carried out in accordance with conventional technical means in the art.

[0022] The anti-aging composition of the present invention comprises polyglutamic acid and its salt, hyaluronic acid and its salt and sweet orange extract.

[0023] Furthermore, the anti-aging composition comprises the following components in parts by weight: 0.01-1.0 parts of polyglutamic acid and its salts, 0.01-1.0 parts of hyaluronic acid and its salts, and 0.0015-1.35 parts of sweet orange extract.

[0024] Furthermore, the anti-aging composition comprises the following components in parts by weight: 0.01-0.3 parts of polyglutamic acid and its salts, 0.01-0.3 parts of hyaluronic acid and its salts, and 0.015-0.9 parts of sweet orange extract.

[0025] For example, the anti-aging composition may contain polyglutamic acid and its salts in an amount of 0.01, 0.02, 0.03, 0.04, 0.05, 0.06, 0.07, 0.08, 0.09, 0.1, 0.15, 0.2, 0.25, 0.3, 0.35, 0.4, 0.45, 0.5, 0.55, 0.6, 0.65, 0.7, 0.75, 0.8, 0.85, 0.9, 0.95, or 1.0 parts by weight.

[0026] The composition may contain hyaluronic acid and its salt in an amount of 0.01, 0.02, 0.03, 0.04, 0.05, 0.06, 0.07, 0.08, 0.09, 0.1, 0.15, 0.2, 0.25, 0.3, 0.35, 0.4, 0.45, 0.5, 0.55, 0.6, 0.65, 0.7, 0.75, 0.8, 0.85, 0.9, 0.95, 1.0, etc. parts by weight.

[0027] The composition may contain 0.0015, 0.002, 0.003, 0.004, 0.005, 0.006, 0.007, 0.008, 0.009, 0.01, 0.015, 0.02, 0.025, 0.03, 0.035, 0.04, 0.045, 0.05, 0.06, 0.07, 0.08, 0.09, 0.10, 0.11, 0.12, 0.13, 0.14, 0.15, 0.16, 0.17, 0.18, 0.19, 0.20, 0.21, 0.22, 0.23, 0.24, 0.25, 0.26, 0.27, 0.28, 0.29, 0.30, 0.31, 0.32, 0.33, 0.34, 0.35, 0.36, 0.37, 0.38, 0.39, 0.40, 0.41, 0.42, 0.43, 0.44, 0.45 .05 part, 0.055 part, 0.06 part, 0.065 part, 0.07 part, 0.075 part, 0.08 part, 0.085 part, 0.09 part, 0.095 part, 0.1 part, 0.2 part, 0.3 part, 0.4 part, 0.5 part, 0.6 part, 0.7 part, 0.8 part, 0.9 part, 1.0 part, 1.1 part, 1.2 parts, 1.3 parts, 1.35 parts, etc.

[0028] In a specific embodiment, the composition comprises, by weight percentage, 0.01-1.0% polyglutamic acid and its salts, 0.01-1.0% hyaluronic acid and its salts, and 0.0015-1.35% sweet orange extract. For example, the polyglutamic acid and its salt can be 0.01%, 0.02%, 0.03%, 0.04%, 0.05%, 0.06%, 0.07%, 0.08%, 0.09%, 0.1%, 0.2%, 0.3%, 0.4%, 0.5%, 0.6%, 0.7%, 0.8%, 0.9%, 1.0%, etc.; the hyaluronic acid and its salt can be 0.01%, 0.02%, 0.03%, 0.04%, 0.05%, 0.06%, 0.07%, 0.08%, 0.09%, 0.1%, 0.2%, 0.3%, 0.4%, 0.5%, 0.6%, 0.7%, 0.8%, 0.9%, 1.0%, etc. 7%, 0.8%, 0.9%, 1.0%, etc.; the sweet orange extract can be 0.0015%, 0.002%, 0.003%, 0.004%, 0.005%, 0.006%, 0.007%, 0.008%, 0.009%, 0.01%, 0.02%, 0.03%, 0.04%, 0.05%, 0.06%, 0.07%, 0.08%, 0.09%, 0.1%, 0.2%, 0.3%, 0.4%, 0.5%, 0.6%, 0.7%, 0.8%, 0.9%, 1.0%, 1.1%, 1.2%, 1.3%, 1.35%, etc.

[0029] Furthermore, the polyglutamic acid and its salts are often used as moisturizers in the cosmetic field and can be purchased directly on the market. The polyglutamic acid (γ-polyglutamic acid, English: Poly-γ-Glutamic acid, referred to as γ-PGA) is polymerized by D-glutamic acid and L-glutamic acid through γ-glutamyl bonds to form a polypeptide macromolecule with a general molecular weight of 10 to 2000 kDa. The polyglutamic acid and its salts in the present application can be any polyglutamic acid, its salt or a combination thereof, including but not limited to polyglutamic acid (γ-PGA, H form), Na + Polyglutamate, K + Polyglutamate in the form of NH4 + in the form of polyglutamate, Mg 2+ in the form of gamma-polyglutamate and / or Ca 2+ Polyglutamate and / or cross-linked γ-polyglutamate hydrogel (made of Na + Polyglutamate, K + Polyglutamate in the form of NH4 + in the form of polyglutamate, Mg 2+ in the form of polyglutamate and / or Ca 2+ In a preferred embodiment of the present application, the polyglutamate is Na + The form of polyglutamate, namely sodium polyglutamate.

[0030] Furthermore, the molecular weight of the polyglutamic acid and its salts of the present application can be any molecular weight that can play the role of the composition of the present application in improving skin elasticity. In a preferred embodiment, the molecular weight is 1.0×10 4 Da-2.0×10 6 Da, for example, can be 1.0×10 4 Da, 5.0×10 4 Da, 1.0×10 5 Da, 1.5×10 5 Da, 2.0×10 5 Da, 2.5×10 5 Da, 3.0×10 5 Da, 4.0×10 5 Da, 5.0×10 5 Da, 6.0×10 5 Da, 7.0×10 5 Da, 8.0×10 5 Da, 9.0×10 5 Da, 1.0×10 6 Da, 1.1×10 6 Da, 1.2×106 Da, 1.3×10 6 Da, 1.4×10 6 Da, 1.5×10 6 Da, 1.6×10 6 Da, 1.7×10 6 Da, 1.8×10 6 Da, 1.9×10 6 Da, 2.0×10 6 Da et al., in one embodiment, the molecular weight is 1.0×10 6 Da-1.8×10 6 Da.

[0031] Further, the hyaluronic acid and its salt refer to a biopolymer material composed of linearly connected repeating units N-acetyl-D-glucosamine and D-glucuronic acid. The hyaluronic acid and its salt include hyaluronic acid itself, its salt or a combination thereof. Hyaluronates include, but are not limited to, inorganic salts such as sodium hyaluronate, potassium hyaluronate, calcium hyaluronate, magnesium hyaluronate, zinc hyaluronate, gold hyaluronate, cobalt hyaluronate; and organic salts such as tetrabutylammonium hyaluronate. In the present application, hyaluronic acid itself or its salt can be used alone, or a combination of two or more hyaluronic acids or their salts can be used.

[0032] The hyaluronic acid and its salt described in the present application are not limited. In a preferred embodiment, the hyaluronate is a water-soluble salt of hyaluronic acid, and is further preferably any one of sodium hyaluronate, zinc hyaluronate, magnesium hyaluronate or potassium hyaluronate. In a specific embodiment, the hyaluronate is sodium hyaluronate. As a natural moisturizing factor, the sodium hyaluronate not only has the effect of moisturizing and hydrating, but also can scavenge free radicals to promote the proliferation and division of epidermal cells and promote the regeneration of skin in injured parts.

[0033] Furthermore, the molecular weight range of the hyaluronic acid and its salts is generally 1.5×10 6 Da or less, for example 5.0×10 3 Da, 1.0×10 4 Da, 5.0×10 4 Da, 1.0×10 5 Da, 2.0×10 5 Da, 3.0×10 5 Da, 4.0×10 5 Da, 5.0×10 5 Da, 6.0×10 5 Da, 7.0×10 5 Da, 8.0×10 5 Da, 9.0×105 Da, 1.0×10 6 Da, 1.1×10 6 Da, 1.2×10 6 Da, 1.3×10 6 Da, 1.4×10 6 Da, 1.5×10 6 Da, preferably 1.0×10 6 Da or less, hyaluronic acid and its salts within this range have similar functions of reducing inflammation, alleviating sensitivity, scavenging free radicals, and remodeling skin barrier, and can all be used in the present invention.

[0034] In some embodiments of the present application, the sweet orange extract can be purchased from the market or extracted by itself. Commercially available products include but are not limited to Logicos sweet orange extract, PROVITAL, SAU sweet orange extract. The self-extraction method is a common method for sweet orange extraction. As long as the extraction method can make the sweet orange extract have conventional efficacy, there is no special restriction. For example, the fruit, leaves, stems or roots of oranges are extracted using a solvent, and the solvent can be water and / or alcohol; the sweet orange extract can also be obtained using callus tissue of orange plants (fruits, leaves, stems or roots).

[0035] In a specific embodiment of the present application, the fruit, leaves, stems or roots of orange are dried in the shade, crushed, dissolved in ethanol, mixed and homogenized in a homogenizer, the homogenized material is placed in an ultrasonic device, an appropriate ethanol solution is added, and ultrasonic extraction is performed to obtain a concentrated solution; the obtained concentrated solution is eluted, the eluate is collected, concentrated, and dried to obtain a sweet orange extract.

[0036] In a specific embodiment of the present application, the orange plant (fruit, leaf, stem or root) can be induced to undergo cell differentiation to obtain callus tissue, and the sweet orange extract can be obtained by cell disruption.

[0037] In a specific embodiment of the present application, the sweet orange extract is 72800-Citrustem purchased from PROVITAL, SAU TM , wherein the content of the active ingredient sweet orange extract is 18%-22%.

[0038] In some embodiments of the present application, the anti-aging composition further comprises auxiliary materials and / or functional ingredients commonly used in the cosmetics field.

[0039] In some embodiments of the present application, the anti-aging composition further comprises excipients, which include one or more of solvents, moisturizers, chelating agents, emollients, skin conditioners, emulsifiers, fragrances, preservatives, thickeners, and pH regulators.

[0040] In some embodiments of the present application, the anti-aging composition further comprises an effective ingredient. The effective ingredient is a commonly used effective ingredient in the art, for example, a moisturizing ingredient, an anti-wrinkle ingredient, a whitening ingredient, an acne-removing ingredient, etc., and those skilled in the art can freely choose.

[0041] Further, in the present application, the effective ingredients may include hyaluronic acid or its derivatives, vitamin C or its derivatives, vitamin A or its derivatives, folic acid or its derivatives, vitamin E or its derivatives, vitamin D or its derivatives, amino acids, carotenoids or carotene, uric acid or its derivatives, α-hydroxy acids, stilbene and its derivatives, tranexamic acid, tetrahydromethylpyrimidine carboxylic acid, dipotassium glycyrrhizinate, glycyrrhetinic acid stearate, vitamin E acetate, vitamin B3, vitamin C, ascorbyl glucoside, sodium vitamin C phosphate, allantoin, peony glycosides, centella asiatica, bisabolol, betaine, sorbitol, sodium pyrrolidonecarboxylate, erythritol, hydrolyzed egg whites, The invention relates to water-soluble extracts of ginseng stem and leaf extract, ginseng seed extract, ginseng extract, bletilla striata extract, white peony extract, polygonatum sibiricum extract, wolfberry extract, peony extract, cistanche deserticola extract, black bean sprout extract, chickpea sprout extract, black sesame extract, black rice extract, aloe vera extract, seaweed extract, algae extract, snow lotus extract, peach blossom extract, green tea extract, seaweed extract, almond extract, pearl hydrolyzate, osmanthus extract, turmeric root extract, lotus extract, ganoderma lucidum extract, koji fermentation-related, plant fermentation product-related, bean fermentation product-related, and fungus-related, one or more of the following.

[0042] The present invention also provides the use of the anti-aging composition in preparing a product for promoting fibroblast proliferation.

[0043] The present invention also provides the use of the anti-aging composition in the preparation of cosmetics, and more preferably in cosmetics for improving skin elasticity and anti-aging cosmetics. The anti-aging composition is added to cosmetics to improve skin elasticity and has the effect of improving the wrinkle removal, firming and anti-aging effects of cosmetics.

[0044] In some embodiments of the present application, the cosmetics include but are not limited to any one of skin care water, skin care essence, skin care lotion, skin care cream, makeup, soap, facial cleanser, shower gel, skin care gel, essence, eye cream, and facial mask.

[0045] In some embodiments of the present application, the composition can be dissolved in an aqueous phase and mixed evenly to obtain an essence. In order to increase the dissolution rate, polyglutamic acid and its salts, hyaluronic acid and its salts and other auxiliary materials and / or effective ingredients can be heated and dissolved before adding the sweet orange extract. There is no specific limitation on the heating temperature as long as the added substances can be dissolved.

[0046] In some embodiments of the present application, the composition can be dissolved in an aqueous phase, and then mixed with an oil phase and homogenized to obtain an emulsion.

[0047] This application does not impose any specific restrictions on the method for preparing cosmetics, and those skilled in the art can choose from commonly used methods for preparing cosmetics.

[0048] In order to better illustrate the technical solutions and advantages of the present invention, the present invention will be further described below in conjunction with specific embodiments. It should be noted that the process parameters and process steps not described in detail in the present invention are all carried out in accordance with conventional technical means in the art.

[0049] In the following examples, polyglutamic acid and its salts, hyaluronic acid and its salts were purchased from Bloomage Biotechnology Co., Ltd., and sweet orange extract was purchased from PROVITAL, SAU (72800-Citrustem TM , sweet orange extract content 18%-22%).

[0050] Example 1

[0051] An anti-aging composition, comprising the following components in parts by weight: 0.1 part of sodium polyglutamate (molecular weight 1.8×10 6 Da), 0.1 part of sodium hyaluronate, molecular weight of sodium hyaluronate 5.0×10 3 Da, sweet orange extract 0.036-0.044 parts.

[0052] Table 1 below shows the compositions of the anti-aging compositions of Examples 1 to 16 and Comparative Examples 1 to 7. A series of anti-aging compositions are obtained according to the formulations in Table 1 below.

[0053] Table 1

[0054]

[0055]

[0056] Comparative Example 8

[0057] An anti-aging composition, comprising the following components in parts by weight: 0.1 part of sodium polyglutamate (molecular weight 1.8×10 6 Da), 0.1 part of sodium hyaluronate, molecular weight of sodium hyaluronate 5.0×10 3 Da, 72800-Citrustem TM 0.2 parts of matrix components excluding active ingredients.

[0058] Application Example 1

[0059] The compositions prepared in the above embodiments and comparative examples can be applied to cosmetics. The types of cosmetics include, but are not limited to, creams, lotions, toners, essences, facial masks, sprays, etc. During preparation, in addition to the composition of polyglutamic acid and its salts, hyaluronic acid and its salts and sweet orange extract, the cosmetics also include a matrix necessary for making various dosage forms. The matrix can be selected from the matrix commonly used for different types of cosmetics according to the prior art, and the cosmetic preparation method can be prepared according to the conventional method of the prior art.

[0060] Application Example 2

[0061] In order to verify the excellent effect of the composition of polyglutamic acid and its salt, hyaluronic acid and its salt and sweet orange extract of the present invention, the composition of the above embodiment and comparative example was added with auxiliary materials to prepare an essence, and the method was as follows:

[0062] Table 2 Essence formula

[0063]

[0064] Table 3 Proportion of ingredients in each embodiment and comparative example in the essence

[0065]

[0066]

[0067]

[0068] The method for preparing the essence is as follows:

[0069] (1) After mixing the raw materials of phase A evenly, start stirring and heat to about 85°C.

[0070] (2) After the solution is evenly dissolved, start cooling.

[0071] (3) When the temperature drops to about 55°C, add the raw materials of phase B and mix evenly to obtain the essence.

[0072] Test 1: Effect on the proliferation of fibroblasts

[0073] Test sample: The essence of Application Example 2 was diluted with ultrapure water at a mass ratio of 2:98 to obtain a 2% test solution.

[0074] Test method: Prepared fibroblast suspension was diluted with 2×10 4The concentration of cells / mL was inoculated in a 96-well plate, 100 μL per well. After the cells adhered to the wall, 20 μL of the above test solution was added to each well with a micropipette. The control group added the same volume of PBS (0.01M), moved into an incubator (under 37°C, 5% carbon dioxide, saturated humidity conditions) and cultured for 24 hours, then added 20 μL of 1 mg / mL MTT solution, and continued to culture under the same conditions for 4 hours. Carefully remove the culture solution in each well, then add 100 μL DMSO to each well and place it on a shaker for 10 minutes at a low speed to fully dissolve the crystals, measure the absorbance A value of each well at OD490nm of the enzyme-linked immunosorbent assay, and set the zero hole (culture medium, MTT, DMSO) at the same time. Calculate the cell survival rate according to the following formula: Cell survival rate = (test sample A value / control group A value) × 100%, with the basic essence formula without adding the anti-aging composition as the control group, and its cell survival rate is 100%. The test results are shown in Table 4.

[0075] Table 4 Effect on proliferation of fibroblasts

[0076]

[0077]

[0078] From the experimental results in Table 4, it can be seen that when the cells are treated with the anti-aging compositions of Examples 1-16, the proliferation of fibroblasts is significantly improved compared with the control examples. In particular, the anti-aging compositions of Examples 1-14 and 16 have a significantly higher proliferation effect on fibroblasts than the control examples. The composition of Example 15 has a significantly higher proliferation effect on fibroblasts than Comparative Examples 1 and 4-7, and Example 15 can achieve similar effects to Comparative Examples 2-3 by using a smaller amount of active ingredients of the anti-aging composition. Although two-row yeast itself has antioxidant and anti-aging effects, it can be seen from Comparative Example 7 that the combination of two-row yeast with sodium polyglutamate and sodium hyaluronate has no obvious proliferation effect on fibroblasts. Compared with Example 1, Comparative Example 8 does not contain sweet orange extract but contains 72800-Citrustem TM The matrix components in Example 8 significantly weakened the proliferation effect of fibroblasts, indicating that it is the sweet orange extract, polyglutamic acid and its salts and hyaluronic acid and its salts, rather than the matrix components, that exert the synergistic anti-aging effect.

[0079] Test 2: Zebrafish embryo reactive oxygen species (ROS) scavenging test

[0080] Test substance treatment group: The test substance treatment group used the essence product prepared according to the essence formula of Table 2 in Example 1;

[0081] Blank control group: the basic essence formula of the anti-aging composition of the present invention is not added.

[0082] Test method: 24 48h zebrafish embryos were exposed to 1g / L solution of the essence of the composition of Example 1, and a blank control group was set up at the same time. After 24h of exposure, the fish embryos were stained with H2DCFDA, and the intensity of ROS signals was measured by fluorescence photography and statistical analysis was performed. The production and removal of reactive oxygen species (ROS) in cells during normal metabolism of human skin remain relatively balanced, but with age, the ability to remove ROS weakens. ROS accumulation accelerates the oxidation and inactivation of proteins, nucleic acids, lipids and other molecules, causing the skin to become loose and wrinkles to form. The ROS regulation mechanism in zebrafish is the same as that in humans, and some fluorescent probes, such as H2DCFDA, can monitor ROS in zebrafish embryos through in vivo staining. Zebrafish embryos are small, which is convenient for taking pictures under a microscope, and the green fluorescence signal in zebrafish embryos that increases with the increase of ROS is quantitatively read by software analysis, and the changes in the ROS levels of fish embryos in the test substance treatment group and the blank control group are tested, and the ROS clearance rate is calculated to evaluate the anti-wrinkle and firming effects of the composition.

[0083] Calculate the ROS clearance rate: clearance rate = (CT) / C×100%

[0084] Where:

[0085] T—the average value of the “average signal intensity” of ROS in the fish embryos of the test substance treatment group;

[0086] C—The average value of the “average signal intensity” of ROS in fish embryos of the blank control group.

[0087] The experimental results are as follows Figure 1 , 2 As shown, from Figure 1 The experimental results show that the brightness area of ​​the test group treated with the anti-aging composition essence of Example 1 is significantly reduced compared with the blank control group, that is, the anti-aging composition essence of the present invention can significantly remove ROS in zebrafish embryos. According to the above clearance rate calculation formula, the test group of the present invention can achieve a zebrafish embryo reactive oxygen species (ROS) clearance rate of 15%. Figure 2 That is, the anti-aging composition essence has an antioxidant effect, can restore skin elasticity, make the skin firm, and improve skin dryness, roughness, and increased wrinkles.

[0088] Test 3: Human efficacy test evaluation

[0089] Test sample: the essence product prepared according to the essence formula of Table 2 in Example 1;

[0090] Blank control group sample: the basic essence formula without adding the anti-aging composition of the present invention.

[0091] Test method: Recruit 30 healthy volunteers, both male and female, aged 20-65 years old.

[0092] Those with the following conditions should be excluded:

[0093] a Those who have used antihistamines in the past week or immunosuppressants in the past month;

[0094] b. Any anti-inflammatory drugs were used in the tested area within the past two months;

[0095] c. Subjects with inflammatory skin diseases that have not been clinically cured;

[0096] dInsulin-dependent diabetic patients;

[0097] e Patients who are receiving treatment for asthma or other chronic respiratory diseases;

[0098] f Those who received anti-cancer chemotherapy in the past 6 months;

[0099] g Patients with immunodeficiency or autoimmune diseases;

[0100] hBreastfeeding or pregnant women;

[0101] i Bilateral mastectomy and bilateral axillary lymph node removal;

[0102] j. The judges whose test results may be affected by scars, pigments, atrophy, port wine stains or other blemishes on the skin to be tested;

[0103] k Participating in other clinical trial researchers;

[0104] lPeople with highly sensitive constitution;

[0105] m Non-voluntary participants or those who cannot complete the required content according to the experimental requirements.

[0106] Volunteers who meet the requirements were recruited, and the half-face control method was adopted. The test samples were used on the left half of the face of the subjects 3 times a week, and the blank control group samples were used on the right half of the face. Skin elasticity was tested 1 week and 2 weeks after use.

[0107] Skin elasticity detection method: Apply constant negative pressure to the skin in the vertical direction, then eliminate the negative pressure, measure the displacement change of the skin in a specified time period, and obtain the skin elasticity characterization value R.

[0108] The calculation formula of skin elasticity characterization value R is:

[0109] Where:

[0110] U a ——The rebound displacement of the skin after the negative pressure is eliminated, in millimeters (mm).

[0111] U f ——The maximum tensile displacement of the skin when negative pressure is applied, in millimeters (mm).

[0112] In the test, the R value is directly calculated and obtained by the instrument (Cutometer Skin Elasticity Tester).

[0113] The relative value of R value Tx vs.T0 means: the ratio of the R value after week X to the R value at week 0 (i.e. before use). This value can directly and accurately express the change of skin elasticity over time. The larger the relative value of R value Tx vs.T0, the better the effect of increasing skin elasticity.

[0114] The results are shown in Table 5 below.

[0115] Table 5 Effects on skin elasticity

[0116]

[0117] According to the results, the volunteers' skin elasticity increased significantly after using the test samples, reaching a peak of 105.94% around the second week of use. The difference between the groups was extremely significant, which means that the scientifically formulated anti-aging composition essence of Example 1 can increase the skin elasticity of the human body.

[0118] Test 4: Human skin patch test

[0119] Test sample: the essence product prepared according to the essence formula of Table 2 in Example 1.

[0120] Test method: Recruit volunteers who meet the requirements. The volunteer conditions are shown in Test 3. The number of volunteers is 30. The test sample is diluted 100 times with purified water and used as a control. Tear open the packaging of the spot tester, measure 0.020ml of the sample, and add it to the small chamber. Apply the spot tester to the flexor side of the subject's forearm, and gently press it with the palm of your hand to evenly apply it to the skin for 24 hours. Observe and record the reaction results according to Table 6 30 minutes, 24 hours and 48 hours after removing the spot tester.

[0121] Table 6 Skin reaction grading standards for skin occlusive patch test

[0122]

[0123] The subjects were observed for skin reactions 30 minutes, 24 hours and 48 hours after removing the spot tester. Results All 30 subjects who were exposed to the anti-aging composition essence of Example 1 had negative reactions. See Table 7 for details.

[0124] Table 7 Statistics of skin reactions

[0125]

[0126] The results of the human skin closed patch test showed that there was no positive reaction in 30 people, which means that the anti-aging composition essence will not cause adverse reactions, is very mild and has no irritation to the human body.

Claims

1. An anti-aging composition, characterized in that: The effective ingredients of the anti-aging composition are composed of the following components in parts by weight: 0.01-1.0 parts of polyglutamic acid and its salts, 0.01-1.0 parts of hyaluronic acid and its salts, and 0.0015-1.35 parts of sweet orange extract; the molecular weight of hyaluronic acid and its salts is 1.5×10 6 Da or less, the molecular weight of polyglutamic acid and its salts is 1.0×10 4 Da-2.0×10 6 Da.

2. The anti-aging composition according to claim 1, characterized in that: The active ingredient is composed of the following components in parts by weight: 0.01-0.3 parts of polyglutamic acid and its salt, 0.01-0.3 parts of hyaluronic acid and its salt, and 0.015-0.9 parts of sweet orange extract.

3. The anti-aging composition according to claim 1, characterized in that: The molecular weight of hyaluronic acid and its salts is 1.0×10 6 Da below.

4. The anti-aging composition according to claim 1 or 2, characterized in that: It also contains auxiliary materials commonly used in the cosmetics field.

5. Use of the anti-aging composition according to any one of claims 1 to 4 in the preparation of a product that promotes fibroblast proliferation.

6. Use of the anti-aging composition according to any one of claims 1 to 4 in the preparation of cosmetics.

7. The use according to claim 6, characterized in that: The anti-aging composition is used for preparing anti-aging cosmetics.

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