Anti-aging composition for activating skin repair mechanism, application and anti-aging skin care product

Abstract

The invention relates to an anti-aging composition for activating a skin repair mechanism, application and an anti-aging skincare product, the composition comprises a bioactive peptide, a silybum marianum extract and a laminaria saccharina extract, and the mass ratio of the bioactive peptide to the silybum marianum extract to the laminaria saccharina extract is (0.01-3): (0.01-2.5): (0.01-3). Compared with the prior art, the functional component provided by the invention can enhance cell viability, promote regeneration of collagen and elastic fibers, relieve damage of oxidative stress to skin cells, and effectively resist skin aging caused by endogenous and exogenous factors.

Description

Technical Field

[0001] The invention belongs to the technical field of daily chemicals and relates to an anti-aging composition for activating skin repair mechanisms and its application, as well as an anti-aging skin care product. Background Art

[0002] Skin aging is a complex and inevitable biological process, manifested by various phenomena such as skin sagging, wrinkle formation, uneven skin tone, dry skin, enlarged pores, and thinning of the skin. These changes are mainly due to the combined effects of endogenous natural aging, oxidation reactions, glycation reactions, photoaging, and unhealthy lifestyle habits and environmental factors. Endogenous natural aging leads to a slowdown in skin cell renewal and an imbalance in the synthesis and degradation of the extracellular matrix (such as collagen and elastic fibers); oxidation reactions and glycation reactions damage the skin structure by producing free radicals and advanced glycation end products (AGEs), respectively, accelerating skin aging; photoaging is mainly caused by ultraviolet radiation, leading to skin photodamage and DNA damage; and unhealthy lifestyle habits and environmental factors, such as smoking, drinking, and pollution, can also have a negative impact on skin health.

[0003] Currently, there are a variety of anti-aging cosmetics on the market, designed to slow the aging process by providing moisturizing, repairing, and antioxidant benefits. However, while traditional anti-aging ingredients such as collagen and hyaluronic acid can improve skin condition to a certain extent, their relatively simple mechanisms of action often hinder their ability to achieve the desired deep-seated skincare effects due to their large molecular weight and poor permeability. This is particularly true in promoting collagen and elastin fiber regeneration, and increasing skin elasticity and firmness. Therefore, the development of more effective, safe, and innovative anti-aging cosmetics has become a key research direction in the industry.

[0004] For example, the anti-aging and firming composition provided by CN202410798779.X includes rice fermentation broth, conoside peptides, and milk thistle extract in a mass ratio of (0.01-30):(0.001-5):(0.001-10). This wide range of ratios and lack of a clear threshold for synergistic effects can easily lead to imbalances in the effects of the ingredients and fluctuations in their effectiveness. Furthermore, the overall solution relies on gel microsphere encapsulation (requiring microfluidics and cross-linking agents), which is complex and costly, and can affect penetration efficiency due to poor particle size control. Summary of the Invention

[0005] The purpose of the present invention is to provide an anti-aging composition and application, as well as an anti-aging skin care product, that activates the skin repair mechanism, can enhance cell vitality, promote the regeneration of collagen and elastic fibers, reduce the damage of oxidative stress to skin cells, and effectively combat skin aging caused by endogenous and exogenous factors.

[0006] The purpose of the present invention can be achieved by the following technical solutions:

[0007] In one aspect, the present invention provides an anti-aging composition for activating skin repair mechanisms, comprising a bioactive peptide, milk thistle extract, and kelp extract, wherein the mass ratio of the bioactive peptide, milk thistle extract, and kelp extract is (0.01-3):(0.01-2.5):(0.01-3). By integrating bioactive peptides such as low-molecular-weight μ-conotoxin peptides with natural extracts, a multi-target repair mechanism is constructed, achieving a synergistic anti-aging effect.

[0008] Furthermore, the bioactive peptide is μ-conotoxin peptide.

[0009] Furthermore, the mass ratio of the μ-conotoxin peptide, milk thistle extract and kelp extract is (0.1-1): (0.1-1.5): (0.1-1).

[0010] The mechanism of action of the anti-aging composition provided by the present invention is:

[0011] Nerve-cell-matrix multi-target combination:

[0012] Neuromodulation: μ-conotoxin peptide, a unique bioactive peptide, primarily targets the neuromuscular junction. By regulating the conduction of nerve signals, μ-conotoxin peptide can effectively reduce wrinkles caused by frequent muscle contractions, thereby improving dynamic lines on the skin.

[0013] Cellular Function Enhancement: As the primary active ingredient at the cellular level, sugar kelp extract is rich in a variety of bioactive substances that promote cell metabolism and anti-oxidation. It activates the antioxidant system within cells, reducing free radical damage to cells. It also stimulates the synthesis of key structural proteins such as collagen, enhancing skin elasticity and firmness.

[0014] Extracellular Matrix Maintenance: Milk thistle extract focuses on protecting the extracellular matrix. By inhibiting the activity of matrix metalloproteinases (MMPs), milk thistle extract can effectively prevent the degradation of extracellular matrix components such as collagen, thereby maintaining the structural integrity and stability of the skin.

[0015] Multi-target synergy: Milk thistle extract, Laminaria japonica extract, and -μ-conotoxin peptide form a synergistic bioactive network. This multi-targeted approach, spanning neuromodulation, cell function promotion, and extracellular matrix maintenance, creates a comprehensive and in-depth anti-aging network. The ingredients complement each other, working together at different levels of the skin to maximize anti-aging benefits.

[0016] The anti-aging composition provided by the present invention contains μ-conotoxin peptide, a short peptide composed of 22 amino acids with a molecular weight between 2000 and 2500 Da. It possesses unique biological activity and highly effective skin repair capabilities. It specifically targets various voltage-sensitive muscle sodium ion channels, particularly the Nav1.4 channel, blocking the influx of Na+ ions within muscle fibers, thereby relaxing facial muscles and instantly reducing the appearance of dynamic wrinkles. Due to its small molecular weight, μ-conotoxin peptide easily penetrates the skin barrier and acts directly on the dermis, improving skin elasticity and firmness.

[0017] Specifically, the mechanism of action of μ-conotoxin peptide in skin cells mainly includes the following aspects:

[0018] (1) Promote the synthesis of extracellular matrix proteins: By activating the TGF-β signaling pathway, it promotes the synthesis of extracellular matrix proteins such as collagen, elastin and hyaluronic acid, thereby increasing the structural support and elasticity of the skin.

[0019] (2) Strengthening the dermal-epidermal junction: The tight connection between the dermis and epidermis is key to maintaining skin stability and firmness. μ-conotoxin peptides enhance skin stability and firmness by promoting the connection between the dermis and epidermis.

[0020] (3) Restore fibroblast vitality: By resetting the function of fibroblasts, it promotes the proliferation, differentiation and migration of skin fibroblasts, restores them to a youthful state, and improves the skin's self-repair and regeneration ability.

[0021] Milk thistle, a herbal remedy with a 2,000-year history of use, is renowned for its seeds in treating liver disease and is a renowned natural remedy. Milk thistle extract, derived from the fruit after the seeds are harvested, is rich in bioactive compounds and exhibits dual biological effects: senolytic (eliminate senescent cells) and senomorphic (remodel senescent cells). Specifically, milk thistle extract selectively induces apoptosis in senescent cells, significantly reducing their negative impact on surrounding healthy cells. It also effectively inhibits the release of harmful factors, including interleukin-6 (IL-6), interleukin-8 (IL-8), and matrix metalloproteinase-1 (MMP-1), by senescent cells, thereby preventing the phenomenon of "contagious aging." Furthermore, milk thistle extract has the ability to reprogram senescent cells, restoring their metabolic function and promoting the synthesis of key skin structural proteins, such as collagen, thereby optimizing skin microstructure. These mechanisms work together to not only effectively eliminate senescent cells but also promote the rejuvenation and functional restoration of skin cells.

[0022] Sugar kelp extract, derived from the marine essence of red algae, is rich in polysaccharides, fucoxanthin, alginate, and other bioactive ingredients. It effectively regulates the skin's water and oil balance, reducing fine lines and wrinkles caused by dryness, and providing lasting moisture and elasticity. Furthermore, its potent antioxidant capacity effectively scavenges free radicals and reduces oxidative stress, delaying the skin's natural aging process at the cellular level and providing deep protection.

[0023] Furthermore, Laminaria japonica extract can promote skin cell metabolism and accelerate the cell renewal cycle, thereby improving uneven skin tone, enhancing skin radiance, and helping to reduce the accumulation of aging cells, maintaining the skin's youthful appearance. Regarding skin repair and regeneration, Laminaria japonica extract can gently reduce the skin's inflammatory response and promote the repair and regeneration of damaged skin tissue.

[0024] In a second aspect, the present invention provides a use of the anti-aging composition for activating the skin repair mechanism in the preparation of anti-aging skin care products.

[0025] Furthermore, the anti-aging skin care products are skin care lotions, essences, lotions, creams, freeze-dried powders, etc., which have the anti-aging effect of activating the skin repair mechanism.

[0026] In a third aspect, the present invention provides an anti-aging skin care product for activating skin repair mechanisms, comprising a matrix excipient and an effective ingredient, wherein the effective ingredient comprises the anti-aging composition as described above, wherein the effective ingredient is added in an amount of 1.2 to 4 wt %. Exemplarily, the effective ingredient can be added in an amount of 1.2 wt %, 1.3 wt %, 1.4 wt %, 1.5 wt %, 1.6 wt %, 1.7 wt %, 1.8 wt %, 1.9 wt %, 2.0 wt %, 2.1 wt %, 2.2 wt %, 2.3 wt %, 2.4 wt %, 2.5 wt %, 2.6 wt %, 2.7 wt %, 2.8 wt %, 2.9 wt %, 3.0 wt % or 4.0 wt %, or any intermediate value within the aforementioned range.

[0027] Furthermore, the matrix excipients include phase A excipients, phase B excipients, phase C excipients, phase D excipients and water. Based on 100 parts by weight of the anti-aging skin care product, the phase A excipients include 0.1 to 10 parts of thickener A, 0.1 to 15 parts of skin conditioner A and 1 to 30 parts of moisturizer A. When the amounts of the components in the phase A excipients are within the above ranges, the fluidity of the product can be improved, making it easier to apply and spread, while maintaining the stability and uniformity of the product.

[0028] Furthermore, the phase B auxiliary material includes 0.1 to 15 parts of excipient B, 0.05 to 10 parts of emulsifier and 0.5 to 5 parts of skin conditioner B.

[0029] Furthermore, the phase C auxiliary material includes 0.1 to 5 parts of a pH adjuster. When the amount of the pH adjuster is within the above range, it is beneficial to maintain the ideal pH range of the product, improve the stability and safety of the product, and improve the user experience of the product.

[0030] Furthermore, the phase D auxiliary material includes 0.1 to 5 parts of a fragrance and 0.01 to 1 parts of a preservative. When the amounts of the components in the phase D auxiliary material are within the above ranges, it can provide a pleasant smell, inhibit the growth and reproduction of microorganisms, and extend the shelf life of the product.

[0031] Furthermore, the water is used to make the weight parts of the anti-aging skin care product reach 100 parts.

[0032] Furthermore, the thickener A is selected from one or more of acrylates / C10-30 alkyl acrylate crosspolymer, polyacrylamide & C13-14 isoparaffin & laureth-7, xanthan gum, cyclopentasiloxane, propylene glycol, behenyl alcohol, carbomer, polyquaternium-37, PVM / MA copolymer, and hydroxyethylcellulose. Exemplary, polyacrylamide, PVM / MA copolymer, and the like can be used.

[0033] Furthermore, the skin conditioning agent A and the skin conditioning agent B are independently selected from one or more of niacinamide, olive oil, sodium hyaluronate, lauryl alcohol, avocado butter, allantoin, jojoba oil, shea butter, hydrogenated polyisobutene, squalane and panthenol.

[0034] Furthermore, the moisturizing agent A includes one or more of hyaluronic acid, glycerin, ceramide, sodium lactate, butylene glycol, hydrogenated grape seed oil and olive oil;

[0035] The excipient B includes one or more of trehalose, polydimethylsiloxane, isohexadecane and glyceryl caprylate.

[0036] Furthermore, the emulsifier includes one or more of olive oil PEG-7 esters, PEG-8 caprylic / capric glycerides, glyceryl stearate, glyceryl stearate citrate, cetearyl glucoside & cetearyl alcohol, behentrimonium methylsulfate and cocoglyceryl;

[0037] The pH regulator includes one or more of arginine and lactic acid;

[0038] The aromatic agent includes essence;

[0039] The preservative includes one or more of phenoxyethanol, ethylhexylglycerin and pentylene glycol.

[0040] Compared with the existing technology, the present invention produces significant anti-aging effects through the anti-aging strategy of "nerve-cell-matrix multi-target combination" and the comprehensive application of core ingredients μ-conotoxin peptide, milk thistle extract and sugar kelp extract, with the following advantages:

[0041] (1) Instantly reduce dynamic wrinkles: μ-conotoxin peptide specifically blocks voltage-dependent sodium channels in muscles, inhibiting muscle contraction, thereby effectively reducing the formation of wrinkles; at the same time, low molecular weight μ-conotoxin peptide (2000-2500Da) and other ingredients can directly penetrate the dermis without the need for complex carriers, resulting in better ingredient permeability and significantly reduced production costs.

[0042] (2) Promote the synthesis and reconstruction of skin extracellular matrix: Microproteins significantly promote the synthesis and reconstruction of skin extracellular matrix components (such as collagen and elastin) by specifically activating the TGF-β (transforming growth factor-β) signaling pathway. Collagen and elastin are key determinants of skin elasticity and firmness. Their increase helps improve skin elasticity and firmness, reducing wrinkles and sagging.

[0043] (3) Antioxidant and anti-inflammatory effects: The natural extracts of the present invention (such as milk thistle extract and kelp extract) are rich in various bioactive ingredients and have significant antioxidant and anti-inflammatory effects. These ingredients can scavenge free radicals, reduce oxidative stress damage to the skin, and inhibit skin inflammatory reactions, thereby protecting the skin from external environmental damage.

[0044] (4) Synergistic Effect: The combination of microproteins and natural extracts produces a significant synergistic effect. Microproteins provide structural support to the skin by promoting cell proliferation and matrix synthesis, while natural plant extracts protect the skin from damage through antioxidant and anti-inflammatory pathways. Through a multi-target mechanism of action, the anti-aging composition of the present invention has a more comprehensive and long-lasting effect.

[0045] Overall, this invention maximizes its anti-aging effects by combining multiple targets to comprehensively improve skin condition at multiple levels, from neuromodulation and cell function promotion to extracellular matrix maintenance. Furthermore, the synergistic effects of the various ingredients enhance the product's overall efficacy and safety, providing users with a comprehensive, long-lasting, and safe skincare solution. BRIEF DESCRIPTION OF THE DRAWINGS

[0046] Figure 1 is the collagen expression level 10D of Example 1, Example 2 and the blank;

[0047] Figure 2 is the mean of the nasolabial fold area test results after using the facial cream of Example 1;

[0048] Figure 3 This is a picture showing the improvement effect of nasolabial folds after using the facial cream of Example 1;

[0049] Figure 4 is the mean of the eye corner wrinkle area test results after using the facial cream of Example 1;

[0050] Figure 5 This is a diagram showing the improvement effect of eye wrinkles after using the facial cream of Example 1;

[0051] Figure 6 It is the mean value of the test results of the moisture content of the stratum corneum after using the facial cream of Example 1. DETAILED DESCRIPTION

[0052] The present invention is described in detail below with reference to the accompanying drawings and specific embodiments. This embodiment is implemented based on the technical solution of the present invention, and provides a detailed implementation method and specific operation process, but the protection scope of the present invention is not limited to the following embodiments.

[0053] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as those commonly understood by those skilled in the art to which this application pertains. The terms used herein in the specification of this application are for the purpose of describing specific embodiments only and are not intended to limit this application.

[0054] The terms "and / or", "or / and", and "and / or" used herein include any one of two or more related listed items, and also include any and all combinations of the related listed items, wherein the any and all combinations include any combination of two related listed items, any more related listed items, or all related listed items. It should be noted that when at least three items are connected by at least two conjunctions selected from "and / or", "or / and", and "and / or", it should be understood that in this application, the technical solution undoubtedly includes technical solutions connected by "logical AND" and technical solutions connected by "logical OR".

[0055] In this application, the technical features described in an open manner include closed technical solutions composed of the listed features, and also include open technical solutions containing the listed features.

[0056] In this application, when referring to numerical ranges, unless otherwise specified, the numerical ranges are considered continuous and include the minimum and maximum values of the range, as well as every value between such minimum and maximum values. Further, when a range refers to an integer, every integer between the minimum and maximum values of the range is included. In addition, when multiple ranges are provided to describe a feature or characteristic, the ranges can be combined. In other words, unless otherwise specified, all ranges disclosed herein should be understood to include any and all subranges subsumed therein.

[0057] Only certain numerical ranges are specifically disclosed herein. However, any lower limit may be combined with any upper limit to form an unspecified range; and any lower limit may be combined with other lower limits to form an unspecified range, and similarly, any upper limit may be combined with any other upper limit to form an unspecified range. In addition, each individually disclosed point or single value may itself serve as a lower limit or upper limit and be combined with any other point or single value, or with other lower limits or upper limits, to form an unspecified range.

[0058] The temperature parameters in this application, unless otherwise specified, allow for both constant temperature treatment and treatment within a certain temperature range. The constant temperature treatment allows for temperature fluctuations within the precision range of instrument control. Fluctuations within ranges such as ±5°C, ±4°C, ±3°C, ±2°C, and ±1°C are permitted.

[0059] Herein, the “suitable” mentioned in “suitable combination”, “suitable method”, “any suitable method”, etc. shall be based on the ability to implement the technical solution of this application, solve the technical problems of this application, and achieve the expected technical effects of this application.

[0060] In this application, "further", "further", "particularly" and the like are used for descriptive purposes to indicate differences in content, but should not be understood as limiting the scope of protection of this application.

[0061] In this application, the terms "optionally," "optional," and "optional" mean optional or dispensable, i.e., they refer to either option being selected from two parallel options: "with" or "without." If a technical solution contains multiple "optional" clauses, each "optional" clause is independent unless otherwise specified and there are no contradictions or constraints.

[0062] In the description of the application, "multiple" means at least two, such as two, three, etc., unless otherwise clearly and specifically defined.

[0063] Unless otherwise stated, all formulations and tests herein took place at 25°C.

[0064] As used herein, the terms "comprise," "include," "contain," "have," "have," or other variations thereof are intended to encompass non-exclusive inclusions, and no distinction is made between these terms. The term "comprising" means that other steps and ingredients that do not affect the end result may be added. The compositions and methods / processes of the present invention comprise, consist of, and consist essentially of the essential elements and limitations described herein, as well as any additional or optional ingredients, components, steps, or limitations described herein. As used herein, no distinction is made between the terms "efficacy," "performance," "effect," and "efficacy."

[0065] Unless otherwise specified, all embodiments and optional embodiments of the present application can be combined with each other to form a new technical solution. Unless otherwise specified, all technical features and optional technical features of the present application can be combined with each other to form a new technical solution.

[0066] Unless otherwise specified, all steps of the present application can be performed sequentially or randomly, preferably sequentially.

[0067] The sugar kelp extract used is derived from the marine biological essence of red algae, which is mainly an extract rich in fucoidan (content ≥ 90wt%). Fucoidan is an important bioactive ingredient in sugar kelp, with significant antioxidant, anti-inflammatory and collagen synthesis promoting effects. In addition, the sugar kelp extract used also contains a certain amount (about 5 to 10wt%) of brown algae polyphenols (Phlorotannins), which further enhances its antioxidant capacity. These ingredients together give it a strong antioxidant capacity and the effect of promoting cell metabolism.

[0068] The rest of the raw materials or processing techniques, unless otherwise specified, are conventional commercially available raw materials or conventional processing techniques that can be used for cosmetics in this field.

[0069] The anti-aging composition for activating the skin repair mechanism provided by the present invention comprises μ-conotoxin peptide, milk thistle extract and kelp extract, wherein the mass ratio of the μ-conotoxin peptide, milk thistle extract and kelp extract is (0.01-3): (0.01-2.5): (0.01-3).

[0070] Preferably, the mass ratio of the μ-conotoxin peptide, milk thistle extract, and kelp extract is (0.1-1): (0.1-1.5): (0.1-1). Exemplarily, the mass ratio of the μ-conotoxin peptide, milk thistle extract, and kelp extract can be 0.5:1:1, 1:1.5:1, 0.2:0.5:0.5, etc., or any other intermediate point or end value within the above-defined mass ratio range.

[0071] To better illustrate the effectiveness of the anti-aging composition provided herein for activating skin repair mechanisms, the following experiments are conducted using a facial cream as an example. However, it should be noted that the anti-aging composition provided herein can be used as an active ingredient and is not limited to facial creams. In addition to the anti-aging composition as an active ingredient, the facial cream may also contain other commonly used base ingredients and excipients.

[0072] Example 1:

[0073] An anti-aging skin care product (face cream) that activates the skin repair mechanism, the proportions of its raw material components are shown in Table 1 below.

[0074] Table 1: Distribution ratio of raw material components in Example 1

[0075]

[0076]

[0077] The preparation process of the anti-aging cream for activating the skin repair mechanism provided in this embodiment is as follows:

[0078] a. Water phase: Place water in a container equipped with a stirring device, fully wet and mix the other parts of the raw material combination A, then add them into the water, stir thoroughly until completely dissolved, and heat to about 75-80℃.

[0079] b. Oil phase: Mix the ingredients in combination B and heat to about 75-80°C, stirring until completely dissolved.

[0080] c. Emulsification: Add the mixture from step b to the aqueous phase from step a, stir at 300-500 rpm for 3-5 minutes and homogenize at 3500-4500 rpm for 3-5 minutes.

[0081] d. Add raw material C, stir at 300-500 rpm for 3-5 minutes, and homogenize at 3000-4000 rpm for 3-5 minutes. Stir and cool to 45-48°C.

[0082] e. Add raw material combination D and stir at 300-500 rpm for 5-10 minutes.

[0083] All the above steps were strictly controlled for sterility.

[0084] f. After cooling to approximately 35-38°C, add combination E, stir at 300-500 rpm for 3-5 minutes, and homogenize at 3000-4000 rpm for 3-5 minutes to obtain the desired cream. Filter and sterilize before filling, and test for bacterial content, toxins, and immunogenicity for quality control.

[0085] Characterization data and effect data of the products of the embodiments and comparative examples

[0086] Examples 2-3, Comparative Examples 1-9 and Blank Example 1

[0087] The only difference between Example 2 and Example 1 is the increase of each active ingredient, and the only difference between Example 3 and Example 1 is the reduction of each active ingredient.

[0088] Comparative Examples 1-3 differ from Example 1 in that only one key composition ingredient is missing, Comparative Examples 4-6 differ from Example 1 in that only two key composition ingredients are missing, and Comparative Example 7 differs from Example 1 in that the key composition ingredients are different in proportion. Comparative Example 8 differs from Example 1 only in that the Laminaria japonica extract in the key composition ingredients is replaced with Centella asiatica extract. Both Centella asiatica extract and Laminaria japonica extract reduce skin oxidative stress through antioxidant and anti-inflammatory pathways and improve skin elasticity by promoting collagen synthesis. Centella asiatica extract and Laminaria japonica extract have similar effects in promoting skin cell metabolism and repairing damaged skin tissue. In Comparative Example 8, Centella asiatica extract is used to replace Laminaria japonica extract to highlight the irreplaceable nature of Laminaria japonica extract in the composition. The difference between Comparative Example 9 and Example 1 is only that the μ-conotoxin peptide in the key composition component is replaced with ω-conotoxin peptide. ω-conotoxin peptide and μ-conotoxin peptide belong to the conotoxin peptide family and are derived from marine cone snail venom. They can affect skin muscle activity by regulating ion channels. The ω-conotoxin peptide has a molecular weight of 3000-3500Da and mainly targets voltage-sensitive calcium ion channels (such as Cav2.2), affecting neurotransmitter release. The μ-conotoxin peptide has a molecular weight of 2000-2500Da, specifically blocks voltage-sensitive sodium ion channels (such as Nav1.4), inhibits muscle contraction, and directly reduces dynamic wrinkles. In Comparative Example 9, ω-conotoxin peptide is used to replace μ-conotoxin peptide for highlighting the irreplaceability of μ-conotoxin peptide in the combination. The difference between Blank Example 1 and Example 1 is only the lack of all key composition components.

[0089] In the following examples, the μ-conotoxin peptide, milk thistle extract, and Centella asiatica extract used are all conventional commercially available cosmetic raw material products.

[0090] The specific ratios are shown in Table 2, and the preparation method is the same as that in Example 1.

[0091] Table 2 Ratio of raw material components of Examples 1-3, Comparative Examples 1-8 and Blank Example 1

[0092]

[0093]

[0094] Characterization data and effect data of the products of the embodiments and comparative examples 1. Safety test

[0095] 1) Test sample: the facial creams of Examples 1-3 and Comparative Examples 1-9 described in Table 2.

[0096] 2) Participants: 130 healthy women aged 30 to 60 years old, all meeting the criteria for the subjects.

[0097] 3) Test Method: 130 subjects who met the test criteria were randomly divided into 13 groups, corresponding to the creams obtained in Examples 1-3 and Comparative Examples 1-9 described in Table 2, and a blank group 1 in which no sample was applied. Each group had 10 subjects; safety was tested by a third-party testing agency. The selected area should not exceed 50mm. 2 Using a suitable patch device with a depth of approximately 1mm, apply approximately 0.02-0.025ml (g) of the test substance to the device using a closed patch test. Apply the patch device to the subject's flexed forearm. After 24 hours, remove the test substance. Observe skin reactions 0.5, 24, and 48 hours after removal. Record the results according to the skin reaction grading standards in the "2015 Cosmetic Safety Technical Regulations."

[0098] Table 3 Patch test results

[0099]

[0100]

[0101] Note: Skin reaction grading standards:

[0102] Grade 0: negative reaction;

[0103] Grade 1: Suspicious reaction; only slight erythema;

[0104] Grade 2: weak positive reaction (erythema reaction); erythema, infiltration, edema, and papules may be present;

[0105] Grade 3: Strong positive reaction (herpes reaction); erythema, infiltration, edema, papules, herpes: the reaction may extend beyond the tested area;

[0106] Grade 4: Very strong reaction (confluent herpes reaction): obvious erythema, severe infiltration, edema, confluent herpes, and the reaction exceeds the test area.

[0107] 3) Test results: The results in Table 3 show no irritation and are considered safe.

[0108] Efficacy evaluation test

[0109] Table 4 Cell experiment raw material composition ratio

[0110]

[0111] 1. Cell proliferation effect test

[0112] Test method:

[0113] 1) Immortalized human epidermal cells (HaCat) were cultured at 10 3 -10 4 The cells were seeded at a density of 100 cells / well in a 96-well plate, 100 μL of culture medium was added, and the cells were cultured at 37°C in a CO2 incubator for 24 hours.

[0114] 2) The samples of Experimental Examples 1-3, Comparative Examples 1-9 and Blank Example 1 were added to the plate and incubated in an incubator for 48 hours.

[0115] 3) Use a repeating pipette to add 10 μL of CCK8 solution to each well of the culture plate and incubate the culture plate in an incubator for 1-4 hours.

[0116] 4) Measure the absorbance at 450 nm using a microplate reader and create a standard curve for data analysis.

[0117] 5) Using blank sample 1 as a reference, calculate the cell proliferation change rate: Cell proliferation change rate (%) = (absorbance of test group - absorbance of blank sample 1) / absorbance of blank sample 1 * 100%. The test results are shown in Table 5 below.

[0118] Table 5 Cell proliferation rate change

[0119] sample Cell proliferation rate (%) Experimental Example 1 45.53 Experimental Example 2 49.38 Experimental Example 3 39.16 Comparative Experimental Example 1 22.39 Comparative Example 2 22.26 Comparative Experiment 3 23.71 Comparative Example 4 8.65 Comparative Experimental Example 5 9.29 Comparative Experimental Example 6 10.02 Comparative Experimental Example 7 37.21 Comparative Experimental Example 8 30.75 Comparative Experimental Example 9 35.18 Experimental blank example 1 0.00

[0120] As shown in Table 5, after cell culture was carried out using the efficacy liquid of Experimental Examples 1-3 and Comparative Examples 1-9, the cell proliferation rate was improved to varying degrees. Among them, the cell proliferation rates of Experimental Examples 1-3 reached 45.53%, 49.38% and 39.16% respectively, and the effect was remarkable. However, the cell proliferation rates of Comparative Examples 1 to 6, which do not contain all the key components (μ-conotoxin peptide, milk thistle extract and kelp extract), varied only between 8.65% and 23.71%, which was at a relatively low level. The proliferation rates of Comparative Example 7, Comparative Example 8 and Comparative Example 9 were 37.21%, 30.75% and 35.18% respectively, which were higher than Comparative Examples 1-6, but still significantly lower than Examples 1-3. This further verifies the unique advantages and synergistic effects of μ-conotoxin peptide, milk thistle extract and kelp extract in the present invention. From the comparison of the results, it can be seen that compared with not using the anti-aging composition with the skin repair mechanism activation, or using only one or two of μ-conotoxin peptide, milk thistle extract and kelp extract, the effect of improving the cell proliferation rate is more obvious when μ-conotoxin peptide, milk thistle extract and kelp extract are used together.

[0121] The key to synergistic efficacy lies in the ratio and selection of the three ingredients within the defined scope of this invention. Experimental data fully confirms that the combination significantly enhances cell viability, promotes proliferation, and delays aging through multi-target synergy, with efficacy significantly superior to that of a control group using a single ingredient, an unrestricted ratio, or alternative ingredients.

[0122] The above experiments show that μ-conotoxin peptide, milk thistle extract and kelp extract have significant synergistic effects. In order to simplify the experimental variables and improve the accuracy and reliability of the research results, only the experimental cases will be analyzed in the subsequent cell experimental research.

[0123] 2. Extracellular matrix protein synthesis effect test

[0124] Test method:

[0125] 1) Immortalized human epidermal cells (HaCat) were cultured at 1x10 4 10 cells / well were seeded in an 8-well plate, and an appropriate amount of complete culture medium (DMEM or RPMI containing 10% FBS) was added to each well to ensure that the cells were evenly distributed.

[0126] 2) Place the culture plate in a CO2 incubator and culture at 37°C, 5% CO2 for 24 hours to allow the cells to adhere to the wall and reach a suitable growth state.

[0127] 3) After the culture, 500 μL of fresh culture medium was taken and added to Experimental Blank Example 1, Experimental Example 1, and Experimental Example 2, respectively. The cells were then cultured for 24 hours and then fixed on each well.

[0128] 4) Measure the expression of collagen using cell immunofluorescence staining.

[0129] The cell color development method is as follows:

[0130] The culture medium was poured off, and the cells were washed three times with PBS for 5 min each time.

[0131] Fix the cells with 4% paraformaldehyde solution at 4°C for 20 minutes. Ensure that the fixative covers the entire cell layer; after fixation, wash the cells three times with pre-chilled PBS for 5 minutes each to remove residual fixative.

[0132] Use a tissue pen to draw circles to prevent the incubation solution from flowing away in the subsequent process, and then rinse with PBS.

[0133] Dilute the antibody (COL3A1) with 5% BSA at a ratio of 1:300, add an appropriate amount of primary antibody (COL3A1) working solution, and incubate at 4°C overnight.

[0134] Remove the culture dish from 4°C and rewarm at room temperature for about 30 minutes. Wash the cells three times with pre-chilled PBS for 5 minutes each to remove unbound primary antibody.

[0135] The antibody (CY3-labeled goat anti-mouse) was diluted 1:100 with 5% BSA, and the secondary antibody (CY3-labeled goat anti-mouse) working solution was added dropwise. The cells were incubated in a 37°C water bath in the dark for 40 minutes. After incubation, the cells were washed three times with pre-cooled PBS for 5 minutes each time to remove unbound secondary antibody.

[0136] DAPI dye was added dropwise to stain the cell nuclei and incubated in the dark at room temperature for 20 to 30 minutes. After staining, the cells were washed three times with pre-cooled PBS for 5 minutes each time to remove excess DAPI dye.

[0137] Mount the cells with anti-fading mounting medium, ensuring that no air bubbles are generated.

[0138] Place the sealed slides under a fluorescence microscope for observation and photography, and record the experimental results.

[0139] Among them, I1 is experimental blank example 1; J1 is experimental example 1; K1 is experimental example 2.

[0140] Depend on Figure 1 It can be seen that the protein expression level in group I1 was at a low level. In contrast, the protein expression levels in groups J1 and K1 were significantly increased. After cell culture using the efficacy solutions of Experimental Examples 1 and 2, the collagen content was significantly increased. This shows that μ-conotoxin peptide, milk thistle extract, and kelp extract can effectively enhance the synthesis of extracellular matrix proteins and increase the structural support and elasticity of the skin.

[0141] 3. Human efficacy test

[0142] (1) Cheek skin firmness F4 test

[0143] Cutometer MPA was used to collect cheek skin firmness;

[0144] 1) Test sample: the facial creams of Examples 1-3, Comparative Examples 1-9 and Blank Example 1 described in Table 2.

[0145] 2) Test subjects: 130 healthy women (aged 30-60) were randomly divided into 13 groups of 10 each. Each group used the same product formula. Written informed consent was obtained. Before enrollment, participants were asked a series of questions regarding their medical history and health status based on the inclusion and exclusion criteria. The skin of the test area was assessed for compliance and skin color was screened, and the results were recorded.

[0146] Environmental Conditions: During the test, visual assessment and instrument testing are conducted in an environment with a temperature of 21±1°C and a relative humidity of 50±10%. Visual assessment is conducted under constant lighting conditions (fluorescent lamps or LED lighting with a color temperature of 5500-6500K). Subjects need to acclimate to these environmental conditions for at least 30 minutes before assessment and testing.

[0147] 3) Testing Method: Use the sample twice daily, morning and evening, for 4 weeks. Apply an appropriate amount of the sample to one cheek and leave the other cheek unused. Apply sunscreen to your entire face every morning.

[0148] The test results were collected at the following three time periods: before using the sample (D0), one week after using the sample (W1), and four weeks after using the sample (W4).

[0149] The test results can be calculated using the following formula:

[0150] Change rate = (analysis value after using the product - analysis value before using the product) ÷ analysis value before using the product × 100%

[0151] 4) Test results

[0152] Cutometer MPA was used to measure cheek skin firmness, and the data can be found in Table 6. A smaller analysis value and a smaller rate of change indicate improved skin firmness.

[0153] Table 6 Change rate of cheek skin firmness

[0154] sample After one week of use % % after four weeks of use Example 1 -3.51 -10.08 Example 2 -3.79 -10.63 Example 3 -3.13 -9.25 Comparative Example 1 -2.45 -7.12 Comparative Example 2 -2.42 -7.08 Comparative Example 3 -2.57 -7.41 Comparative Example 4 -2.08 -4.36 Comparative Example 5 -2.13 -5.48 Comparative Example 6 -2.16 -5.64 Comparative Example 7 -3.02 -8.87 Comparative Example 8 -2.89 -8.14 Comparative Example 9 -2.97 -8.86 Blank example 1 -1.04 -0.85

[0155] The results in Table 6 show that after using the creams of Examples 1-3 and Comparative Examples 1-9, the firmness of the cheek skin of the subjects showed an improvement over time. After four weeks of use, the changes in Examples 1-3 were -10.08%, -10.63%, and -9.25%, respectively, demonstrating significant improvements. This demonstrates that the anti-aging composition of the present invention can effectively enhance skin firmness, reduce sagging, and improve the skin's structural support and elasticity.

[0156] Specifically, after one week of use, the creams of Examples 1-3 showed changes in cheek skin firmness of -3.51%, -3.79%, and -3.13%, respectively, demonstrating a modest improvement observed even in the initial phase of use. With extended use to four weeks, the improvement further intensified, with changes exceeding -9%, demonstrating the cumulative and long-lasting effects of the compositions of the present invention over time.

[0157] In contrast, the creams of Comparative Examples 1-6 showed a firmness change rate of only -4.36% to -7.41% after four weeks of use, significantly lower than the Example group. This indicates that the lack of the key ingredients of the present invention (μ-conotoxin peptide, milk thistle extract, and kelp extract) or the use of only one or two of these ingredients cannot achieve the same firmness improvement effect, further validating the synergistic effect of the ingredients in the present composition.

[0158] The firmness changes of the creams in Comparative Examples 7, 8, and 9 after four weeks of use were -8.87%, -8.14%, and -8.86%, respectively. While higher than those in Comparative Examples 1-6, they were still lower than those in Examples 1-3. This indicates that even with partial replacement ingredients (such as Centella asiatica extract or ω-conotoxin peptide), it is impossible to completely replace the key ingredients in the present invention and achieve the same firmness-improving effect.

[0159] The firmness of the cheek skin in Blank Example 1 (no active ingredient used) did not change significantly, which further confirms that the anti-aging effect of the experimental cream is not natural or due to other external factors.

[0160] From the comparison of the results of Examples 1-3, Comparative Examples 1-9 and Blank Example 1, it can be seen that compared with the anti-aging composition that does not use an anti-aging composition that activates the skin repair mechanism, or only uses one or more of μ-conotoxin peptide, milk thistle extract and sugar kelp extract, when μ-conotoxin peptide, milk thistle extract and sugar kelp extract are used at the same time and the ratio of the present invention is followed, the synergistic effect is significantly better than that of the control group with a single ingredient, an unrestricted ratio or an alternative ingredient, and the improvement effect of cheek skin firmness is better, and there is an anti-aging effect. Example 1 is a basic formula that contains all key ingredients and preliminarily verifies its anti-aging effectiveness. In order to simplify the experimental variables and improve the accuracy and reliability of the research results, only Example 1 is analyzed in the subsequent human efficacy test.

[0161] (2) Nasal fold area test

[0162] The skin fast optical imaging system EvaFACE (EO-TECH, USA) is used to measure facial wrinkles.

[0163] 1) Test sample: the facial cream of Example 1 described in Table 2.

[0164] 2) Test subjects: 33 healthy women (aged 30-60 years). Written informed consent was obtained. Prior to enrollment, a series of questions regarding medical history and health status were asked based on inclusion and exclusion criteria. Conformity assessment and skin color testing of the test area were also performed, and the results were recorded.

[0165] Environmental Conditions: During the test, visual assessment and instrument testing are conducted in an environment with a temperature of 21±1°C and a relative humidity of 50±10%. Visual assessment is conducted under constant lighting conditions (fluorescent lamps or LED lighting with a color temperature of 5500-6500K). Subjects need to acclimate to these environmental conditions for at least 30 minutes before assessment and testing.

[0166] 3) Testing Method: Use the sample twice daily, morning and evening, for 4 weeks. Apply an appropriate amount of the sample to one cheek and leave the other cheek unused. Apply sunscreen to your entire face every morning.

[0167] The test results were collected at the following three time periods: before using the sample (D0), one week after using the sample (W1), and four weeks after using the sample (W4).

[0168] The test results can be calculated using the following formula:

[0169] Change rate = (analysis value after using the product - analysis value before using the product) ÷ analysis value before using the product × 100%

[0170] 4) Test results

[0171] The descriptive analysis of the test results is shown in Table 7. The trend of change is shown in Table 7. Figure 2 As shown, the improvement effect is as Figure 3 shown.

[0172] Table 7 Descriptive analysis of nasolabial fold area test results

[0173]

[0174] The results showed that compared with before using the sample, the area of nasolabial folds decreased significantly by 4.63% (p<0.001) after 1 week of use, and further decreased to 6.60% after 4 weeks (p<0.001), indicating that the anti-aging effect of the product gradually accumulates and enhances with the extension of usage time.

[0175] (3) Eye wrinkle area test

[0176] The skin rapid optical imaging system EvaFACE (EO-TECH, USA) was used to measure the area of eye wrinkles;

[0177] Parameter explanation: The lower the value, the smaller the wrinkle area in the test area. Unit: mm 2 .

[0178] 1) Test sample: the facial cream of Example 1 described in Table 2.

[0179] 2) Test subjects: 33 healthy women (aged 30-60 years). Written informed consent was obtained. Prior to enrollment, a series of questions regarding medical history and health status were asked based on inclusion and exclusion criteria. Conformity assessment and skin color testing of the test area were also performed, and the results were recorded.

[0180] Environmental Conditions: During the test, visual assessment and instrument testing are conducted in an environment with a temperature of 21±1°C and a relative humidity of 50±10%. Visual assessment is conducted under constant lighting conditions (fluorescent lamps or LED lighting with a color temperature of 5500-6500K). Subjects need to acclimate to these environmental conditions for at least 30 minutes before assessment and testing.

[0181] 3) Testing Method: Use the sample twice daily, morning and evening, for 4 weeks. Apply an appropriate amount of the sample to one eye area, and leave the other eye area unused. Apply sunscreen to the entire face every morning.

[0182] The test results were collected at the following three time periods: before using the sample (D0), one week after using the sample (W1), and four weeks after using the sample (W4).

[0183] The test results can be calculated using the following formula:

[0184] Change rate = (analysis value after using the product - analysis value before using the product) ÷ analysis value before using the product × 100%

[0185] 4) Test results

[0186] The skin fast optical imaging system EvaFACE (EO-TECH, USA) was used to measure the wrinkle area at the corner of the eye. The descriptive analysis of the test results is shown in Table 8. The change trend is shown in Table 8. Figure 4 and Figure 5 shown.

[0187] Table 8 Test results and statistical analysis of eye wrinkle area

[0188]

[0189] The results showed that compared with before using the sample, the area of wrinkles at the corners of the eyes was significantly reduced by 9.06% (p<0.001) after 1 week of use, and further reduced to 11.39% after 4 weeks (p<0.001), indicating that the anti-aging effect of the product gradually accumulates and enhances with the extension of usage time.

[0190] (3) Test of moisture content of stratum corneum

[0191] Corneometer CM825 is used to collect stratum corneum moisture content;

[0192] 1) Test sample: the facial cream of Example 1 described in Table 2.

[0193] 2) Test subjects: 33 healthy women (aged 30-60 years). Written informed consent was obtained. Prior to enrollment, a series of questions regarding medical history and health status were asked based on inclusion and exclusion criteria. Conformity assessment and skin color testing of the test area were also performed, and the results were recorded.

[0194] Environmental Conditions: During the test, visual assessment and instrument testing are conducted in an environment with a temperature of 21±1°C and a relative humidity of 50±10%. Visual assessment is conducted under constant lighting conditions (fluorescent lamps or LED lighting with a color temperature of 5500-6500K). Subjects need to acclimate to these environmental conditions for at least 30 minutes before assessment and testing.

[0195] 3) Testing Method: Use the sample twice daily, morning and evening, for 4 weeks. Apply an appropriate amount of the sample to one cheek and leave the other cheek unused. Apply sunscreen to your entire face every morning.

[0196] The test results were collected at the following three time periods: before using the sample (D0), one week after using the sample (W1), and four weeks after using the sample (W4).

[0197] The test results can be calculated using the following formula:

[0198] Change rate = (analysis value after using the product - analysis value before using the product) ÷ analysis value before using the product × 100%

[0199] 4) Test results

[0200] Corneometer CM825 was used to collect the moisture content of the stratum corneum. The test was repeated 6 times and the average value was obtained. The descriptive analysis of the test results is shown in Table 9. The change trend is shown in Table 9. Figure 6 shown.

[0201] Table 9 Descriptive analysis of stratum corneum moisture content test results

[0202]

[0203]

[0204] The results showed that compared to before using the sample, the moisture content of the cheek stratum corneum increased significantly by 15.91% after one week of use (p<0.001), and further increased to 36.53% after four weeks (p<0.001). This shows that the sample can quickly replenish water and maintain skin moisture for a long time, effectively alleviating dry skin.

[0205] The above description of the embodiments is intended to facilitate understanding and use of the invention by those skilled in the art. It will be apparent that those skilled in the art can readily make various modifications to these embodiments and apply the general principles described herein to other embodiments without requiring inventive effort. Therefore, the present invention is not limited to the above-described embodiments. Improvements and modifications made by those skilled in the art based on the disclosure of the present invention without departing from the scope of the present invention should be within the scope of protection of the present invention.

Claims

1. An anti-aging composition for activating skin repair mechanism, characterized in that: The invention comprises bioactive peptides, milk thistle extract and kelp extract, wherein the mass ratio of the bioactive peptides, milk thistle extract and kelp extract is (0.01-3): (0.01-2.5): (0.01-3).

2. The anti-aging composition for activating skin repair mechanism according to claim 1, characterized in that: The bioactive peptide is μ-conotoxin peptide.

3. The anti-aging composition for activating skin repair mechanism according to claim 2, characterized in that: The mass ratio of the μ-conotoxin peptide, milk thistle extract and kelp extract is (0.1-1): (0.1-1.5): (0.1-1).

4. Use of the anti-aging composition for activating the skin repair mechanism according to any one of claims 1 to 3 in the preparation of anti-aging skin care products.

5. The use of the anti-aging composition for activating skin repair mechanism according to claim 4, characterized in that: The anti-aging skin care products are skin care lotions, essences, emulsions, face creams, and freeze-dried powders.

6. An anti-aging skin care product that activates the skin repair mechanism, characterized in that: The invention comprises a matrix auxiliary material and an effective component, wherein the effective component comprises the anti-aging composition according to any one of claims 1 to 3, wherein the added amount of the effective component is 1.2 to 4 wt%.

7. The anti-aging skin care product for activating the skin repair mechanism according to claim 6, characterized in that: The matrix excipients include phase A excipients, phase B excipients, phase C excipients, phase D excipients and water. Based on 100 parts by weight of the anti-aging skin care product, the phase A excipients include 0.1 to 10 parts of thickener A, 0.1 to 15 parts of skin conditioner A and 1 to 30 parts of moisturizer A. The phase B auxiliary material includes 0.1 to 15 parts of excipient B, 0.05 to 10 parts of emulsifier and 0.5 to 5 parts of skin conditioner B; The phase C auxiliary material includes 0.1 to 5 parts of a pH regulator; The phase D auxiliary material includes 0.1 to 5 parts of fragrance and 0.01 to 1 parts of preservative; The water is used to make the weight parts of the anti-aging skin care product reach 100 parts.

8. The anti-aging skin care product for activating the skin repair mechanism according to claim 7, characterized in that: The thickener A is selected from one or more of acrylic acid (esters) / C10-30 alkyl acrylate crosspolymer, polyacrylamide & C13-14 isoparaffin & laureth-7, xanthan gum, cyclopentasiloxane, propylene glycol, behenyl alcohol, carbomer, polyquaternium-37, PVM / MA copolymer and hydroxyethyl cellulose; The skin conditioning agent A and the skin conditioning agent B are independently selected from one or more of niacinamide, olive oil, sodium hyaluronate, lauryl alcohol, avocado butter, allantoin, jojoba oil, shea butter, hydrogenated polyisobutene, squalane and panthenol.

9. The anti-aging skin care product for activating the skin repair mechanism according to claim 7, characterized in that: The moisturizing agent A comprises one or more of hyaluronic acid, glycerin, ceramide, sodium lactate, butylene glycol, hydrogenated grape seed oil and olive oil; The excipient B includes one or more of trehalose, polydimethylsiloxane, isohexadecane and glyceryl caprylate.

10. The anti-aging skin care product for activating the skin repair mechanism according to claim 7, characterized in that: The emulsifier includes one or more of olive oil PEG-7 esters, PEG-8 caprylic / capric glycerides, glyceryl stearate, glyceryl stearate citrate, cetearyl glucoside & cetearyl alcohol, behentrimonium methosulfate and cocoglyceryl; The pH regulator includes one or more of arginine and lactic acid; The aromatic agent includes essence; The preservative includes one or more of phenoxyethanol, ethylhexylglycerin and pentylene glycol.

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