Composition for targeted improvement of dark fine wrinkles of sensitive skin as well as preparation and application of composition

Abstract

The invention discloses a composition for improving dark fine lines of sensitive skin in a targeted manner as well as preparation and application of the composition, and belongs to the technical field of cosmetics. The composition for improving the dark and fine wrinkles of the sensitive skin in a targeted manner, provided by the invention, is prepared from the following components in parts by mass: 0.01 to 10 parts of oxyresveratrol, 0.01 to 2 parts of fibronectin, 0.01 to 2 parts of carnosine and 0.01 to 3 parts of carrot seed oil. According to the invention, oxyresveratrol, fibronectin, carnosine and carrot seed oil in a specific mass part range are selected and compounded, so that a good inhibition effect on tyrosinase, TNF-alpha and IL-1alpha can be effectively realized, and meanwhile, an excellent DPPH (1, 1-diphenyl-2-picrylhydrazyl) removal effect can also be realized; when the composition is added into cosmetics subsequently, good whitening, anti-inflammatory and anti-wrinkle effects can be achieved.

Description

Technical Field

[0001] This invention belongs to the field of cosmetic technology, and in particular relates to a composition for targeting and improving dullness and fine lines on sensitive skin, its preparation and application. Background Technology

[0002] Current cosmetics often focus on optimizing single ingredients, neglecting the synergistic effect of multiple active ingredients, and lack dynamic response design to the skin's microecology (such as pH and redox), making it difficult to balance product efficacy stability and user experience.

[0003] In existing technologies, achieving multiple effects requires addressing the compatibility of various active ingredients and studying their interactions. Anti-inflammatory, whitening, and anti-wrinkle ingredients, needed to improve dullness and fine lines in sensitive skin, generally have significantly different physicochemical properties within the same formulation, easily leading to instability and performance degradation. Furthermore, many multifunctional products suffer from low transdermal absorption rates; traditional formulations' active ingredients, due to their molecular size and polarity, struggle to penetrate the skin barrier and achieve effective concentrations at the target site. For example, some large-molecule peptide anti-wrinkle ingredients and plant extracts have difficulty penetrating the stratum corneum, limiting their actual efficacy. Simultaneously, existing technologies struggle to achieve the optimal balance and synergistic enhancement of whitening, anti-inflammatory, and anti-wrinkle effects; most products tend to focus on one or two effects, failing to truly achieve a synergistic effect across all three. For instance, excessive pursuit of whitening effects may damage the skin barrier, triggering inflammation and exacerbating sensitive skin symptoms, while excessive addition of anti-inflammatory ingredients may interfere with the effectiveness of whitening and anti-wrinkle ingredients, failing to improve dullness and fine lines.

[0004] Therefore, it is urgent to develop products that simultaneously offer excellent whitening, anti-inflammatory, and anti-wrinkle effects. Summary of the Invention

[0005] The purpose of this invention is to overcome the shortcomings of the prior art and provide a composition that targets and improves dullness and fine lines on sensitive skin, which has excellent whitening, anti-inflammatory and antioxidant effects, as well as its preparation and application.

[0006] To achieve the above objectives, in a first aspect of the present invention, the present invention provides a composition for targeting and improving dullness and fine lines on sensitive skin, the composition comprising the following components in parts by weight: 0.01-10 parts of oxidized resveratrol, 0.01-2 parts of fibronectin, 0.01-2 parts of carnosine, and 0.01-3 parts of carrot seed oil.

[0007] This invention has found that by selecting components within the above-mentioned mass fraction range for compounding, it is possible to effectively inhibit tyrosinase, TNF-α and IL-1α, while also achieving excellent DPPH clearance. When subsequently added to cosmetics, it can achieve good whitening, anti-inflammatory and anti-wrinkle effects.

[0008] Specifically, oxidized resveratrol, as an active derivative of resveratrol, is rich in active groups such as phenolic hydroxyl groups in its molecular structure. These groups endow it with strong antioxidant efficacy and stability. From the perspective of its mechanism of action, oxidized resveratrol can achieve anti-inflammatory effects by regulating inflammation-related signaling pathways in the body and inhibiting the release of inflammatory factors TNF-α and IL-1α. Simultaneously, it can also inhibit tyrosinase activity, reducing melanin production and thus achieving a skin-whitening effect.

[0009] Fibronectin, as a core component of the extracellular matrix, has a unique tripeptide repeat sequence structure. This structure enables it to specifically bind to integrin receptors on the cell surface. With its properties of promoting cell migration and activating collagen synthesis, it effectively increases the collagen content in the skin and strengthens the skin's supporting structure, thus effectively reducing wrinkles and improving skin laxity and wrinkle problems.

[0010] As a natural antioxidant, carnosine has a unique electron distribution in its imidazole ring structure, which allows it to neutralize free radicals by providing electrons and terminate the chain reaction initiated by free radicals. At the same time, carnosine can also bind to sugar molecules, inhibit glycation reactions, reduce the formation of advanced glycation end products (AGEs), effectively reduce yellowing of the skin and improve skin elasticity, so that the skin can regain firmness and radiance.

[0011] Carrot seed oil is rich in natural antioxidants such as carotenoids and vitamin E. Carotenoids can be converted into vitamin A in the body, which can regulate the proliferation and differentiation of skin cells and promote skin metabolism. Vitamin E can directly capture free radicals and protect cell membranes from free radical damage. These components in carrot seed oil work synergistically to help reduce the risk of cell damage from free radicals, achieving anti-wrinkle effects on multiple levels.

[0012] Through extensive experimentation, the inventors creatively discovered that combining the four components in specific mass proportions achieves excellent synergistic effects. They hypothesize that, regarding the synergistic effect between components, the phenolic hydroxyl groups in oxidized resveratrol and the amino groups in carnosine may exhibit hydrogen-bonded interactions. This interaction enhances the stability of both components in the system while promoting their penetration and absorption in the skin. This allows them to overcome their individual limitations (e.g., oxidized resveratrol has poor solubility and is prone to photolysis, making it difficult to balance stability and absorption for efficacy; carnosine also has stability issues), thus more effectively exerting antioxidant and anti-inflammatory effects. The macromolecular structure of fibronectin provides an excellent carrier platform for oxidized resveratrol and carnosine, distributing them evenly on the skin surface and within the skin, increasing their contact with skin cells, thereby enhancing overall efficacy. The fat-soluble components in carrot seed oil effectively improve the compatibility between the components. Because resveratrol oxide, fibronectin, and carnosine exhibit different solubilities in solvents, carrot seed oil can serve as a good solvent and dispersant, allowing these components to mix uniformly and form a stable complex system. Furthermore, some components of carrot seed oil may promote skin absorption, effectively enhancing the absorption of other components, especially the relatively large molecular weight fibronectin. This helps open channels in the stratum corneum, allowing other components to better penetrate deeper into the skin, further enhancing the overall effect of the composition.

[0013] The four components selected in this invention, through a rational combination, not only achieve synergistic effects but also produce effects that individual components cannot achieve. For example, this composition can simultaneously achieve excellent inhibitory effects on tyrosinase, TNF-α, and IL-1α. Oxidized resveratrol mainly inhibits tyrosinase and inflammatory factors, while fibronectin and carnosine synergistically enhance the regulation of inflammation and melanin production by oxidized resveratrol from the perspectives of promoting cell repair and antioxidation. At the same time, the composition also achieves excellent DPPH scavenging effects. Each component scavenge free radicals through different pathways, complementing each other and greatly improving the overall antioxidant capacity. Therefore, when subsequently applied to cosmetics, the resulting cosmetics have excellent comprehensive effects, comprehensively improving skin condition and meeting consumers' diverse needs for beauty and skincare.

[0014] As a preferred embodiment of the composition of the present invention, the composition comprises the following components in parts by weight: 0.1-5 parts of oxidized resveratrol, 0.01-1 parts of fibronectin, 0.05-1 parts of carnosine, and 0.1-2 parts of carrot seed oil.

[0015] As a preferred embodiment of the composition of the present invention, the composition comprises the following components in parts by weight: 0.5-1 parts of oxidized resveratrol, 0.3-0.5 parts of fibronectin, 0.1-0.2 parts of carnosine, and 0.5-1 parts of carrot seed oil.

[0016] The present invention has found that when the components of the composition are further selected within the above-mentioned range, the overall performance of the resulting composition is even better.

[0017] In a preferred embodiment of the composition of the present invention, the composition contains 8-38% carrot seed oil by mass.

[0018] For example, the mass percentage of carrot seed oil in the composition can be any point value or a range between any two points between 8% and 38%, such as 8%, 10%, 12%, 14%, 16%, 18%, 20%, 22%, 24%, 26%, 28%, 30%, 32%, 34%, 36%, 38%, etc.

[0019] Preferably, the composition contains 24-38% carrot seed oil by mass.

[0020] The present invention has found that when the mass percentage of carrot seed oil in the composition is further controlled within the above-mentioned range, the resulting composition exhibits better inhibitory effects on tyrosinase, TNF-α, and IL-1α, and also better DPPH scavenging effects.

[0021] As a preferred embodiment of the composition of the present invention, the carrot seed oil has an acid value ≤3.0 mg KOH / g and a saponification value of 3-10 mg KOH / g.

[0022] Preferably, the carrot seed oil has an acid value of 0.2-1.5 mg KOH / g and a saponification value of 3-6 mg KOH / g.

[0023] It should be noted that the test method for the acid value of the carrot seed oil is as follows: Accurately weigh approximately 5g of carrot seed oil sample (accurate to 0.001g) and place it in a 250mL Erlenmeyer flask. Add 50mL of neutral ethanol (95% ethanol, neutral as determined by phenolphthalein indicator) and heat to 50℃ to completely dissolve the sample. Add 3-4 drops of phenolphthalein indicator and titrate with 0.1mol / L potassium hydroxide ethanol solution until the solution turns pink and remains so for 30 seconds. Record the volume of potassium hydroxide solution consumed (V, unit: mL).

[0024] Blank experiment: Perform a blank experiment (without adding a sample) following the same steps, and record the volume (V, unit: mL) of potassium hydroxide solution consumed in the blank.

[0025] Acid value calculation formula: Acid value = ((VV0) × c × 56.1) / m in: c: Concentration of potassium hydroxide solution (mol / L); m: Sample mass (g); 56.1: Molar mass of potassium hydroxide (g / mol).

[0026] The test method for saponification value is as follows: Accurately weigh approximately 2g of carrot seed oil sample (accurate to 0.001g) and place it in a 250mL alkali-resistant conical flask. Add 25mL of 0.5mol / L potassium hydroxide ethanol solution (potassium hydroxide purity ≥85%, ethanol concentration 95%) and connect a reflux condenser. Heat under reflux for 60 minutes to ensure complete saponification of the oil. After cooling, add 1mL of phenolphthalein indicator and titrate with 0.5mol / L hydrochloric acid standard solution until the red color disappears. Record the volume of hydrochloric acid solution consumed (V, unit: mL).

[0027] Blank experiment: Perform the blank experiment (without adding sample) following the same steps, and record the volume (V) of hydrochloric acid solution consumed in the blank experiment. (Unit: mL)

[0028] Formula for calculating saponification value (mg KOH / g): Saponification value = (56.1 × (V0V1) × c) / m in: c: Concentration of hydrochloric acid solution (mol / L); m: Sample mass (g); 56.1: Molar mass of potassium hydroxide (g / mol).

[0029] For example, the acid value of the carrot seed oil can be any point value or any range between any two points between ≤3.0 mg KOH / g, such as 1 mg KOH / g, 2 mg KOH / g, 3 mg KOH / g, etc.; the saponification value can be any point value or any range between any two points between 3-10 mg KOH / g, such as 3 mg KOH / g, 5 mg KOH / g, 7 mg KOH / g, 9 mg KOH / g, 10 mg KOH / g.

[0030] This study found that selecting carrot seed oil with acid value and saponification value within the above-mentioned range allows it to interact better with other components in the composition, thereby enhancing the overall effect of the composition.

[0031] As a preferred embodiment of the composition of the present invention, the method for preparing the carrot seed oil includes the following steps: pulverizing and sieving dried carrot seeds, then steam distilling them, collecting the condensed oil phase and distilling it under reduced pressure to obtain carrot seed oil.

[0032] The present invention has found that further selection of the carrot seed oil preparation method provided by the present invention can better achieve the excellent comprehensive effect of the composition of the present invention.

[0033] Preferably, the moisture content of the dried carrot seeds is ≤10%.

[0034] Preferably, the sieving is done through a 20-40 mesh sieve.

[0035] Preferably, the distillation time is 4-6 hours.

[0036] Preferably, the condensation temperature is 25-30°C.

[0037] Preferably, the temperature of the vacuum distillation is ≤60℃ and the pressure is ≤1kPa.

[0038] In a second aspect, the present invention provides a whitening, anti-inflammatory, and antioxidant nanovesicle, the nanovesicle comprising a core and a wall, the core comprising the composition described in the present invention, and the wall comprising Tween-80, propylene glycol, and polyethylene glycol-polypropylene sulfide (PEG-b-PPS). The mass ratio of Tween-80, propylene glycol, and polyethylene glycol-polypropylene sulfide is (0.01-5):(1-15):(0.01-5).

[0039] This invention has discovered that by further forming the composition into the nanovesicles described in this invention, on the one hand, the bilayer membrane structure of the vesicles encapsulates the composition within an internal aqueous phase or hydrophobic interlayer, forming a physical barrier that effectively prevents direct contact and degradation of the active substances in the composition by external environments (such as enzymes, pH changes, oxidants, etc.), thereby improving the stability of the composition; on the other hand, the bilayer membrane structure of the vesicles has a high degree of similarity to the lipid bilayer of the stratum corneum of the skin, enabling it to closely contact and fuse with the stratum corneum, allowing the encapsulated composition to directly penetrate the stratum corneum barrier and enhancing the skin permeability of the composition.

[0040] Specifically, the nanovesicles provided by this invention have a vesicle wall comprising Tween-80, propylene glycol, and polyethylene glycol-polypropylene sulfide within a specific mass ratio range. This allows them to utilize a redox response mechanism. When the nanovesicles enter a high ROS environment (such as an inflammatory site), the sulfide groups (-S-) in the vesicle wall structure are oxidized to sulfoxides (-SO-) or sulfones (-SO2-) under the action of ROS (such as oxides like H2O2), leading to increased hydrophilicity of the copolymer, vesicle structure destruction, and release of the vesicle core. This enables precise release of the composition in inflammatory or other high ROS environments, exerting anti-inflammatory, melanin-inhibiting (tyrosinase activity inhibition rate ≥80%), and free radical scavenging (DPPH scavenging rate ≥90%) effects. Therefore, when subsequently applied to cosmetics, this can effectively improve the efficacy and gentleness of the cosmetics.

[0041] As a preferred embodiment of the nanovesicles of the present invention, the mass ratio of Tween-80, propylene glycol, and polyethylene glycol-polypropylene sulfide is (0.1-1):(3-10):(0.1-1).

[0042] The present invention has found that when the mass ratio of the capsule wall components is further selected within the above range, the average particle size of the obtained nanovesicles is within a suitable range, and the structural stability is better, the encapsulation efficiency is higher, and the permeation-promoting effect of the nanovesicles obtained by the general formula is also better.

[0043] In a preferred embodiment of the nanovesicles described in this invention, the block ratio of polyethylene glycol to polypropylene sulfide in the polyethylene glycol-polypropylene sulfide is (1-2):1.

[0044] In a preferred embodiment of the nanovesicles of the present invention, the mass percentage of the core in the nanovesicles is 20-36%. For example, it can be 20%, 22%, 24%, 26%, 28%, 30%, 32%, 34%, 36%, etc.

[0045] In a preferred embodiment of the nanovesicles of the present invention, the average particle size of the nanovesicles is 150-300 nm.

[0046] As a preferred embodiment of the nanovesicles of the present invention, the method for preparing the nanovesicles includes the following steps: (1) Tween-80, propylene glycol and polyethylene glycol-polypropylene sulfide were dissolved in chloroform, and then oxidized resveratrol and carrot seed oil were added and dissolved by ultrasonication to obtain an organic phase; (2) Dissolve fibronectin and carnosine in preheated PBS buffer to obtain the aqueous phase; (3) After removing chloroform by rotary evaporation of the organic phase, the aqueous phase is added for hydration, followed by sonication, ultrafiltration, centrifugation, collection of centrifugal sediment and freeze-drying to obtain nanovesicles.

[0047] Preferably, in step (2), the preheating temperature is 37±2℃.

[0048] Preferably, in step (2), the ratio of the sum of the masses of fibronectin and carnosine to the volume of PBS buffer is (0.4-0.7) g: 100 mL.

[0049] Preferably, in step (3), the rotary evaporation temperature is 40-50℃ and the rotation speed is 50-150rpm.

[0050] Preferably, in step (3), the ultrasound is performed with a power of 180-200W for 3-7 seconds, with an interval of 8-12 seconds, and repeated 4-6 times.

[0051] Preferably, in step (3), the centrifugation speed is 4000-6000 rpm and the time is 25-35 min.

[0052] Preferably, in step (3), the freeze-drying process involves pre-freezing to -40±5℃ and then heating to 20-30℃ for 30-42 hours.

[0053] A third aspect of the invention provides the use of the composition or nanovesicles described herein in the preparation of cosmetics.

[0054] As a preferred embodiment of the application described in this invention, the cosmetic includes any one of toner, lotion, gel, mask, serum, gel, spray, and cream.

[0055] In a fourth aspect, the present invention provides a skin care gel comprising the composition described herein or the nanovesicles.

[0056] As a preferred embodiment of the skin care gel of the present invention, the skin care gel comprises the following components by weight percentage: 2-7.5% composition, 0.05-0.8% thickener, 0.5-10% moisturizer, 0.5-20% oil, 0.5-5% emulsifier, 0.5-3% preservative, 0.01-0.3% pH adjuster, and the balance being water.

[0057] As a preferred embodiment of the skin care gel of the present invention, the skin care gel comprises the following components by weight percentage: 10-20% nanovesicles, 0.05-0.8% thickener, 0.5-10% moisturizer, 0.5-20% oil, 0.5-5% emulsifier, 0.5-3% preservative, 0.01-0.3% pH adjuster, and the balance being water.

[0058] Preferably, the thickener includes at least one of xanthan gum, carbomer, polyacrylamide taurate, ammonium acryloyldimethyl taurate / VP copolymer, and sclerotium gum.

[0059] Preferably, the moisturizer includes at least one of allantoin, propylene glycol, panthenol, β-glucan, trehalose, caprylyl glycol, dipropylene glycol, sodium hyaluronate, 1,3-butanediol, glycerin, bud break enzyme polysaccharide, and ceramide.

[0060] Preferably, the pH adjuster includes at least one of arginine and tromethamine.

[0061] Preferably, the oil comprises at least one of squalane, isononyl isononanoate, caprylic / capric triglyceride, polydimethylsiloxane, jojoba oil, grape seed oil, shea butter, sunflower seed oil, and vitamin E.

[0062] Preferably, the emulsifier comprises at least one of cetearyl alcohol, glyceryl stearate, polyglycerol-10 diisostearate, cetearyl alcohol polyether-20, C12-20 alkyl glucoside, cetearyl glucoside, polysorbate-80, pentaerythritol tetraester, and sucrose stearate.

[0063] Preferably, the preservative includes at least one of 1,2-propanediol, 1,2-hexanediol, ethylhexylglycerin, p-hydroxyacetophenone, capryloyl hydroxamic acid, and phenoxyethanol.

[0064] Compared with the prior art, the beneficial effects of the present invention are as follows: This invention, by selecting a specific mass range of resveratrol oxidized, fibronectin, carnosine, and carrot seed oil for compounding, can effectively achieve good inhibitory effects on tyrosinase, TNF-α, and IL-1α, while also achieving excellent DPPH scavenging effects; when subsequently added to cosmetics, it can achieve good whitening, anti-inflammatory, and anti-wrinkle effects. Attached Figure Description

[0065] Figure 1 A before-and-after photograph of the wrinkle area of ​​volunteers using the skin care gel prepared in Application Example 1 in Effect Example 5; Figure 2 The image shows a before-and-after comparison of inflammation and whitening in volunteers using the skincare gel prepared in Example 1, as shown in Example 5. Detailed Implementation

[0066] To better illustrate the purpose, technical solution, and advantages of the present invention, the present invention will be further described below in conjunction with specific embodiments.

[0067] Unless otherwise specified, the reagents, methods and equipment used in this invention are all conventional reagents, methods and equipment in the field; and unless otherwise specified, the raw materials used in parallel experiments are from the same batch.

[0068] Fibronectin 1: Plant-derived fibronectin, manufactured by Wuhan Heyuan Biotechnology Co., Ltd., product number: Plant-derived fibronectin OsrFN liquid; Fibronectin 2: Fibronectin from bovine plasma, manufactured by Guangzhou Lucheng Biotechnology Co., Ltd., product number 341631; Carnosine: manufactured by Jiangsu Chengxin Pharmaceutical Co., Ltd., product name CarnoShine. TM L-carnosine; Carrot seed oil 1: Homemade, the preparation method includes the following steps: (1) Raw material pretreatment: Select mature and dry carrot seeds (seeds of carrot, a plant of the Apiaceae family), and avoid using raw materials that are moldy or have too many impurities. Rinse the carrot seeds with clean water to remove surface dirt and impurities, and place them in a ventilated place to dry naturally until the moisture content is ≤10%. Then, crush the dried carrot seeds to 20 mesh to obtain crushed carrot seeds. (2) Steam distillation extraction: Use a steam distillation apparatus, including a distillation pot, condenser, oil-water separator, etc. Check the equipment for airtightness to avoid steam leakage during distillation. Put the crushed carrot seeds into the distillation pot, filling it to 60% of the pot's volume. Add distilled water twice the weight of the crushed carrot seeds. Then heat the pot initially until the water boils (100℃). Distill at atmospheric pressure. After the steam stabilizes, maintain the temperature of the distillation pot between 100-105℃ and the temperature of the condenser between 25-30℃. Continue distilling for 5 hours until no more oil droplets are separated from the oil-water separator. The distilled mixture enters the oil-water separator. After standing for 30 minutes, separate the upper oil layer and dry it with anhydrous sodium sulfate to remove residual water to obtain the crude product. (3) Refining and storage: The crude product is further removed by vacuum distillation (temperature ≤60℃, pressure ≤1kPa). After refining, the carrot seed oil is a light yellow transparent liquid with no suspended matter, an acid value of 0.2mg KOH / g, and a saponification value of 3mg KOH / g.

[0069] Carrot seed oil 2: Homemade. The preparation method differs from that of carrot seed oil 1 in that the temperature of the distillation pot is maintained at 110-115℃ and the distillation time is 8 hours. The resulting carrot seed oil has an acid value of 3 mg KOH / g and a saponification value of 9 mg KOH / g.

[0070] Carrot seed oil 3: Homemade, the preparation method includes the following steps: The difference between the preparation method and carrot seed oil 1 is that after separating the upper oil layer, step (3) is carried out directly, that is, the step of drying with anhydrous sodium sulfate to remove residual water is not used. Carrot seed oil is obtained, which is a light yellow transparent liquid with no suspended matter, an acid value of 4 mg KOH / g and a saponification value of 12 mg KOH / g.

[0071] Carrot Seed Oil 4: Homemade, prepared by cold pressing. Mature, dried carrot seeds (seeds of the carrot plant, Apiaceae family) are selected, avoiding the use of moldy or excessively impurity-laden raw materials. The carrot seeds are washed clean with water to remove surface dirt and impurities, and placed in a ventilated place to dry naturally until the moisture content is ≤10%. Then, the dried carrot seeds are crushed to 20 mesh to obtain crushed carrot seeds. The crushed carrot seeds are then pressed at a pressure of 40MPa and a temperature of 45℃ for 10-20 minutes per batch, with 3 pressing batches. After standing, the oil and water are separated. The oil is filtered through a vacuum oil filter using a 200-mesh filter cloth. The resulting carrot seed oil has an acid value of 1.2mg KOH / g and a saponification value of 5.3mg KOH / g.

[0072] Examples 1-9 and Comparative Examples 1-4 The present invention provides a whitening, anti-inflammatory, and antioxidant composition in the embodiments and comparative examples. The components (parts by mass) of the whitening, anti-inflammatory, and antioxidant composition are shown in Table 1, wherein W represents the mass percentage of carrot seed oil in the composition. Table 1 The composition provided in Example 1 is prepared by mixing the components evenly to obtain the composition.

[0073] The preparation methods of the compositions provided in Examples 2-9 and Comparative Examples 1-4 are consistent with those in Example 1; if the relevant components are not available, they can be omitted.

[0074] Comparative Example 5 The present invention provides a composition in a comparative example, the difference between the composition and Example 1 being that resveratrol is used instead of oxidized resveratrol.

[0075] Comparative Example 6 The present invention provides a composition in a comparative example, the difference between the composition and Example 1 being that epidermal growth factor is used instead of fibronectin.

[0076] Comparative Example 7 The present invention provides a composition in a comparative example, the difference between the composition and Example 1 being that positronic acid is used instead of carnosine.

[0077] Comparative Example 8 The present invention provides a composition in a comparative example, the difference between the composition and Example 1 being that sea buckthorn seed oil (purchased from Shaanxi Guozhen Biotechnology Co., Ltd.) is used instead of carrot seed oil.

[0078] Example 10 This invention provides a nanovesicle, the preparation method of which includes the following steps: (1) Dissolve 0.5g Tween-80, 6g propylene glycol and 0.3g PEG-b-PPS (in PEG-b-PPS, the block ratio of polyethylene glycol and polypropylene sulfide is 1:1, the manufacturer is Xi'an Qiyue Biotechnology Co., Ltd., the product number is Q-0197188) in 100mL chloroform at a mass ratio of 1.7:20:1. Then add oxidized resveratrol and carrot seed oil 1 according to the mass parts of the composition in Example 1, and sonicate until completely dissolved to obtain the organic phase; (2) Fibronectin 1 and carnosine were dissolved in 99.7 mL of PBS buffer preheated to 37 °C to obtain an aqueous phase; wherein the ratio of the total mass of the composition to the total mass of the capsule wall components (Tween-80, propylene glycol and PEG-b-PPS) was 1:2.5. (3) Transfer the organic phase to a round-bottom flask and remove chloroform by rotary evaporation (45°C, 100 rpm) to form a uniform film. Then add the aqueous phase and gently shake until the film is completely hydrated. Transfer the hydrated solution to a centrifuge tube and centrifuge briefly to remove air bubbles and undissolved impurities. Place the centrifuged solution in an ultrasonic instrument and sonicate (power 200W, sonicate for 5 seconds, interval 10 seconds, repeat 5 times) to form a vesicle solution. (4) Select a 50kDa ultrafiltration tube, transfer the ultrasonically treated vesicle solution into the ultrafiltration tube, and perform ultrafiltration centrifugation at 5000rpm for 30min. After centrifugation, the vesicles will remain in the ultrafiltration tube, while the unencapsulated active substances and organic solvents will pass through the filter membrane into the filtrate. Wash the vesicles in the ultrafiltration tube with an appropriate amount of deionized water, and centrifuge again to remove residual impurities. After centrifugation, transfer the purified vesicle solution from the ultrafiltration tube to a freeze-drying bottle, and place it in a freeze dryer for freeze-drying under the following conditions: pre-freeze to -40℃, then heat to room temperature (25℃), and dry for 36h to obtain nanovesicles.

[0079] Examples 11-18 This invention provides a nanovesicle. The preparation method of the nanovesicle differs from that of Example 10 in that the composition is different. Examples 11-18 correspond to the compositions in Examples 2-9, respectively. That is, Example 11 corresponds to the composition in Example 2, Example 12 corresponds to the composition in Example 3, and so on.

[0080] Example 19 This invention provides a nanovesicle, the preparation method of which differs from that of Example 10 in that the mass ratio of Tween-80, propylene glycol, and polyethylene glycol-polypropylene sulfide is 0.1:10:1.

[0081] Example 20 This invention provides a nanovesicle, the preparation method of which differs from that of Example 10 in that the mass ratio of Tween-80, propylene glycol, and polyethylene glycol-polypropylene sulfide is 1:3:0.1.

[0082] Example 21 This invention provides a nanovesicle, the preparation method of which differs from that of Example 10 in that the mass ratio of Tween-80, propylene glycol, and polyethylene glycol-polypropylene sulfide is 0.01:15:5.

[0083] Example 22 This invention provides a nanovesicle, the preparation method of which differs from that of Example 10 in that the mass ratio of Tween-80, propylene glycol, and polyethylene glycol-polypropylene sulfide is 5:1:0.01.

[0084] Comparative Examples 9-16 The present invention provides a nanovesicle in a comparative example. The preparation method of the nanovesicle differs from that of Example 10 in that the composition is different. Comparative Examples 9-16 correspond to the compositions in Comparative Examples 2-8, respectively. That is, Comparative Example 9 corresponds to the composition in Comparative Example 2, Comparative Example 10 corresponds to the composition in Comparative Example 3, and so on.

[0085] Comparative Example 17 The present invention provides a nanovesicle in a comparative example. The preparation method of the nanovesicle differs from that of Example 10 in that the mass ratio of Tween-80, propylene glycol, and polyethylene glycol-polypropylene sulfide is 0.01:1:10.

[0086] Comparative Example 18 The present invention provides a nanovesicle in a comparative example. The preparation method of the nanovesicle differs from that of Example 10 in that the mass ratio of Tween-80, propylene glycol, and polyethylene glycol-polypropylene sulfide is 5:15:0.005.

[0087] Comparative Example 19 The present invention provides a nanovesicle in a comparative example. The difference between the preparation method of the nanovesicle and that of Example 10 is that polyethylene glycol-polypropylene glycol copolymer is used instead of polyethylene glycol-polypropylene sulfide.

[0088] Application examples, comparison application examples, and blank application examples The present invention provides a skin care gel in application examples, comparative application examples, and blank application examples. The components (mass percentage) of the skin care gel are shown in Table 2. In application examples 1-13, the nanovesicles used are those prepared in Examples 10-22, for example, application example 1 uses the nanovesicles from Example 10, application example 2 uses the nanovesicles from Example 11, and so on. In comparative application examples 1-11, the nanovesicles used are those prepared in Comparative Examples 9-19. In application example 14, the nanovesicles used are those prepared in Example 10. Application Examples 15-16 use the compositions prepared in Example 1; Table 2 The preparation method of the skin care gel provided in Example 1 includes the following steps: (1) Mix the humectant and thickener with water and stir. Heat to 75±2℃ and homogenize at 1200rpm for 4min. After homogenization, keep warm for later use to obtain pre-prepared component A. (2) Mix the emulsifier with the oil and heat it to 75±2℃ to obtain the pre-prepared component B; (3) Mix the preservatives and heat to 60±2℃ to melt them to obtain pre-prepared component C; (4) Heat the pre-prepared component A to 80±2℃, add the pre-prepared component B (heated to 80±2℃) at 1200 rpm, homogenize, then cool to 60±2℃, add the pre-prepared component C at 250 rpm and stir to mix, then cool to 40℃ and add the whitening, anti-inflammatory and antioxidant composition and continue stirring for 8 minutes, finally add the remaining pH adjuster to adjust the pH to 6.0, stop stirring, discharge the product, and obtain the skin care gel.

[0089] The preparation methods of the skin care gels provided in Application Examples 2-16, Comparative Application Examples 1-10, and Blank Application Examples are consistent with those in Application Example 1. If the relevant components are not available, they can be omitted.

[0090] Example of effect 1 The effects of this invention are investigated in Examples 10, 19-22, and Comparative Examples 17-19, examining the average particle size, encapsulation efficiency, redox responsiveness, and structural changes of the nanovesicles prepared therein. Specifically, the following are included: 1. Average particle size test The particle size of nanovesicles was determined using dynamic light scattering (DLS). Specifically, 200 μL of nanovesicle solution (distilled water, concentration 1.0 μg / mL) was taken and a nanoparticle size analyzer (WINNER-801) was used. The detection angle was set to 90°, the photon correlator received the light signal, the temperature was 25℃ (constant temperature control), the equilibration time was 120 seconds (to ensure sample temperature stability), and the measurement was repeated three times for each sample, with 300 data points measured each time to improve statistical accuracy. The software automatically calculated the average particle size and recorded it in Table 3.

[0091] 2. Encapsulation efficiency test Encapsulation efficiency was determined using an ultrafiltration tube centrifugation method, employing a 15 mL MWCO 10k ultrafiltration tube. The nanovesicle solution (9.5% concentration, volume ≤15 mL) was added to the inner tube of the ultrafiltration tube, the tube was capped, and after equilibration with an empty ultrafiltration tube, it was centrifuged at 3000 rpm for 30 min. After centrifugation, free drug was present in the filtrate of the outer tube, while the nanovesicles were retained in the inner tube. The drug concentrations in the inner tube (encapsulated drug) and outer tube (free drug) were measured separately, and the encapsulation efficiency was calculated as follows: Encapsulation efficiency (%) = (Drug concentration in inner tube / (Drug concentration in inner tube + Drug concentration in outer tube)) × 100%. The results are recorded in Table 3.

[0092] 3. Redox responsiveness verification Take 1 mL of nanovesicle suspension (solution: PBS, nanovesicle concentration approximately 3 mg / mL) and transfer it to an MWCO 3500 dialysis bag. Immerse the dialysis bag in 50 mL of PBS buffer (pH 7.4) and pre-equilibrate at 37°C with constant shaking (100 rpm) for 30 min. Add H2O2 to the external solution of the dialysis bag to a final concentration of 1 mM (simulating ROS levels under pathological conditions). Set up a control group: no H2O2 added, only PBS buffer. At time points of 0, 4, 12, and 24 h, take 100 μL of sample from the external solution of the dialysis bag and simultaneously add 100 μL of fresh PBS (to maintain a constant volume). After filtering through a 0.22 μm filter membrane, the sample is used for HPLC to detect drug concentration. (2) Drug release rate detection (HPLC method) Preparation of standard curve: Accurately weigh oxidized resveratrol standard, dissolve and dilute with methanol to a series of concentrations (0.1-100 μg / mL), analyze by HPLC, and plot the peak area-concentration standard curve (R2≥0.999); Sample detection: Take 100 μL of the external dialysis bag sample, add 900 μL of methanol, vortex mix, centrifuge at 12000 rpm for 10 minutes, and inject the supernatant into HPLC to detect the peak area of ​​oxidized resveratrol (detection wavelength 306 nm). Calculate the drug concentration based on the standard curve, and calculate the cumulative release rate according to the following formula: Where: C n V: Concentration of the drug in the dialysis bag fluid at the nth sampling time (μg / mL); C: Total volume of the dialysis bag fluid (usually 50 mL, which needs to be kept constant); i : The concentration of the drug in the dialysis bag fluid at the time of the i-th sampling (μg / mL); V i : The volume of liquid removed during the i-th sampling (usually 1 mL); M total The total amount of drug loaded in the nanovesicles (μg) is shown in Table 3.

[0093] 4. Structural change monitoring The particle size of nanovesicles was determined using dynamic light scattering (DLS). Specifically, at time points of 0, 4, and 12 hours during redox response verification, 100 μL of vesicle suspension from the dialysis bag was taken, diluted to 1 mL with PBS, and thoroughly mixed. A WINNER-801 nanoparticle size analyzer was used, with the detection angle set to 90°, a photon correlator receiving the light signal, a temperature of 25℃ (thermostatic control), an equilibration time of 120 s (to ensure sample temperature stability), and measurements repeated three times per sample. 300 data points were measured each time to improve statistical accuracy. The software automatically calculated the average particle size. Subsequently, the rate of change of the average particle size at 4 hours and 12 hours compared to 0 hours was calculated. The results are shown in Table 3. Table 3 As can be seen from Table 3, when the technical solution provided by this invention is adopted, the obtained nanovesicles have a moderate average particle size, a high encapsulation efficiency, and excellent redox responsiveness and structural stability. Specifically, the average particle size of the obtained nanovesicles is between 160±8 and 289±10 nm, and the encapsulation efficiency is between 61% and 85%. In the redox responsiveness test, the cumulative release rates at 4h, 12h, and 24h are above 6±0.8%, 18±2.0%, and 25±2.8%, respectively. In the structural change monitoring, the change rates of the average particle size at 4h and 12h compared to 0h are above 25% and 50%, respectively. As can be seen from Examples 10, 19-22, and Comparative Examples 17-18, changing the selection of the capsule wall components resulted in different average particle size, encapsulation efficiency, redox responsiveness, and structural changes during the redox response process of the obtained nanovesicles. When the proportion of the capsule wall components in Comparative Examples 17-18 was outside the scope of this invention, the encapsulation efficiency of the obtained nanovesicles decreased significantly, the redox responsiveness also decreased significantly, and the rate of structural change during the test also decreased significantly. The presumed reason is that the proportion of the capsule wall components affects the formation mode and structural density of the capsule wall. When the component proportions are within a suitable range, the components can arrange and interact in an orderly manner in a specific way to form a complete and dense capsule wall. This dense structure can effectively encapsulate the internal composition, reduce its leakage, and thus improve the encapsulation efficiency. For example, if the proportion of a component with good film-forming properties (such as Tween-80) is too low, it may lead to a loose capsule wall structure with more pores, making it easy for the encapsulated material to escape, and the encapsulation efficiency will decrease significantly. Conversely, if the proportion of this component is too high, although the capsule wall may be denser, excessive accumulation may affect the flexibility of the capsule wall and its compatibility with the internal substances, which is also detrimental to improving the encapsulation efficiency. Furthermore, the distribution and concentration of redox functional groups are closely related to the component ratio. When the component ratio is appropriate, redox-sensitive groups can be uniformly distributed in the capsule wall and reach a suitable concentration, allowing the nanovesicles to undergo rapid chemical reactions upon encountering redox stimuli, leading to capsule wall structure destruction and drug release. When the ratio changes, the distribution of sensitive groups may be uneven or the concentration too low, thus reducing the sensitivity of the nanovesicles to redox stimuli and significantly decreasing the responsiveness. The proportion of capsule wall components affects the electronic structure and electron transport capacity of the capsule wall; a suitable component ratio can give the capsule wall good electronic conductivity, which is beneficial for redox reactions. During the redox response, the structural changes of the capsule wall involve not only chemical reactions but also mechanical properties. The component ratio affects the mechanical strength and toughness of the capsule wall. When the capsule wall has appropriate mechanical properties, it can undergo gradual and orderly structural destruction under redox stimuli, achieving controllable drug release. If the component ratio leads to poor capsule wall mechanical properties, such as insufficient strength or toughness, sudden rupture or uneven structural changes may occur during the response process, affecting the structural change rate and the stability of drug release. In Comparative Example 17, the mass of Tween-80 and propylene glycol is relatively low, while the mass of polyethylene glycol-polypropylene sulfide is relatively high. This may mean that the redox-sensitive groups cannot be uniformly distributed and reach a suitable concentration within the capsule wall, resulting in poor encapsulation efficiency and redox responsiveness. Therefore, under the premise of stable vesicle structure, the vesicle is less likely to be triggered by redox reactions to release the internal encapsulated material, and the structural change is relatively small. In Comparative Example 18, the mass of Tween-80 and propylene glycol is relatively high, while the mass ratio of polyethylene glycol-polypropylene sulfide is very low (0.005), resulting in poor redox responsiveness and relatively small structural changes.

[0094] As can be seen from Examples 10 and 19, when the capsule wall component is not the substance described in this invention, the average particle size and encapsulation efficiency of the obtained nanovesicles change less, but the redox responsiveness deteriorates, and the rate of structural change in the redox responsiveness test also decreases significantly. This is because in Comparative Example 19, polyethylene glycol-polypropylene glycol copolymer was used instead of polyethylene glycol-polypropylene sulfide. Polyethylene glycol-polypropylene glycol copolymer does not possess redox responsiveness, therefore its redox responsiveness is worse than in Example 10, and the structural change is significantly smaller. Other components remained consistent with Example 10, hence the smaller changes in average particle size and encapsulation efficiency.

[0095] Example 2 The transdermal permeability of the nanovesicles prepared in Examples 10, 19-22, and Comparative Examples 17-19 was investigated using the following steps: (1) Preparation of test samples: The nanovesicles prepared in Examples 10, 19-22 and Comparative Examples 17-19 were prepared into corresponding skin care gels using the preparation method and components as described in Application Example 1, wherein the mass of oxidized resveratrol in the skin care gel was controlled to be 20 mg, and test samples were obtained. (2) Using the Franz diffusion cell, the exposed skin area in the diffusion cell was 1.5 cm². 2 The receiving chamber volume was 5.0 mL. Using rat skin as a barrier, 2 mL of test sample was added to the donor chamber and physiological saline to the receiving chamber. By precisely controlling the skin surface temperature (25℃ conventional / 37℃ response), 0.5 mL samples were taken from the receiving chamber at 0.5, 1, 4, 8, 12, and 24 h (with an equal volume of fresh physiological saline added simultaneously). The cumulative resveratrol permeation was quantitatively analyzed by HPLC, with the unit of cumulative permeation being μg / cm³. 2 The results are shown in Table 4. Table 4 As can be seen from Table 4, when the nanovesicle scheme provided by this invention is selected, the obtained nanovesicles exhibit good transdermal permeability, with cumulative permeation amounts of 26 μg / cm³ at 4h, 12h, and 24h. 2 Above, 70μg / cm 2 Above, 100μg / cm 2 above; As can be seen from Examples 10, 19-22, and Comparative Examples 17-18, the transdermal permeability of the obtained nanovesicles varies when the selection of the capsule wall components is changed. When the mass ratio of the capsule wall components in Comparative Examples 17-18 is outside the range given in this invention, the transdermal permeability of the obtained product decreases significantly. This is partly because particle size affects permeability; relatively speaking, smaller particle sizes are easier to penetrate the skin. Another reason is that differences in redox response capabilities affect the ability to release contents during transdermal penetration. Therefore, the particle size of Comparative Example 17 is relatively larger than that of Example 10, and the proportion of the capsule wall composition is also outside the range given in this invention. Within the scope of this invention, its redox responsiveness is poor, thus significantly reducing the transdermal permeability. The proportion of the capsule wall composition in Comparative Example 18 is also outside the scope of this invention, resulting in poor redox responsiveness and a significant decrease in transdermal permeability. As can be seen from Examples 10 and 19, when the capsule wall component is not a substance of this invention, the transdermal permeability of the obtained nanovesicles is significantly reduced. This is because although the particle size of Comparative Example 19 is not much different from that of Example 10, Comparative Example 19 does not possess redox responsiveness. Therefore, in actual transdermal permeation, it is difficult to induce the release of the internal encapsulation, thus significantly reducing the amount of oxidized resveratrol permeated.

[0096] Example 3 This invention investigates the effects of nanovesicles in vitro. Since cell experiments typically use static culture systems, they lack the influence of the dynamic skin environment on the distribution of active ingredients. At the animal or human level, nanovesicles may promote skin penetration of the encapsulated substance, thus enhancing its efficacy. However, in cell experiments, nanovesicles are directly dissolved and destroyed, releasing the core composition. Therefore, in vitro experiments cannot demonstrate the advantages of this nanovesicle dosage form. Thus, this invention investigates the performance of the compositions prepared in Examples 1-9 and Comparative Examples 1-8, as well as the capsule wall composition formed from Tween-80, propylene glycol, and polyethylene glycol-polypropylene sulfide in a mass ratio of 0.5:6:0.3, including the following aspects: 1. In vitro anti-inflammatory test (1) Reagent preparation Culture medium preparation: Prepare DMEM culture medium containing 10% (V / V) fetal bovine serum (FBS).

[0097] Trypsin-EDTA solution: Aliquot the 0.25% trypsin-EDTA solution into small volumes (8 mL), freeze at -20°C, and thaw quickly in a 37°C water bath before use to avoid repeated freeze-thaw cycles affecting activity.

[0098] Lipopolysaccharide (LPS): Prepare a stock solution of 1 mg / mL LPS using sterile PBS, aliquot and store at –20°C. For experiments, dilute with culture medium to a working solution of 1 μg / mL according to group requirements, preparing fresh each time.

[0099] Sample solutions: The compositions of Examples 1-9 and Comparative Examples 1-8, as well as the capsule wall composition, were dissolved in DMSO to prepare stock solutions of 10 mg / mL and stored at -20°C. Before use, the solutions were diluted with culture medium to a working concentration of 100 μg / mL, ensuring the final DMSO concentration did not exceed 0.1% to eliminate any potential impact on the cells.

[0100] Dexamethasone: Prepare a 10 mM stock solution of dexamethasone using DMSO and store at -20°C. For experiments, dilute with culture medium to a 1 μM working solution and prepare fresh each time.

[0101] ELISA kit reagents: Prepare the necessary washing buffer, detection antibody, standards, and other reagents according to the instructions of the selected TNF-α and IL-1α inflammatory factor ELISA kit, and bring them to room temperature in advance.

[0102] (2) Consumables preparation Cell culture consumables: Prepare T75 culture flasks, 96-well cell culture plates, 15mL and 50mL centrifuge tubes, pipette tips (10μL, 100μL, 200μL, 1000μL), cell scrapers, etc. Ensure that all consumables are sterile products and use them as soon as possible after opening.

[0103] Other consumables: Prepare 96-well plate sealing film, aluminum foil (for protecting reagents from light), ice box (for keeping samples at low temperature during operation), etc.

[0104] (3) Instrument preparation Cell culture related instruments: Turn on the CO2 incubator in advance, adjust the temperature to 37℃, CO2 concentration to 5%, and humidity to above 95%, and clean and disinfect it. Pre-cool the centrifuge to 4℃ for cell centrifugation.

[0105] Testing instruments: Preheat the ELISA reader and calibrate and test its performance according to the instruction manual to ensure the accuracy of the test results. At the same time, prepare instruments such as vortex mixers and micropipettes, and calibrate and check their functionality.

[0106] (4) Cell Culture RAW264.7 mouse macrophages were seeded in T75 culture flasks and cultured in DMEM medium containing 10% FBS at 37°C and 5% CO2. When the cells reached the logarithmic growth phase, they were digested with 0.25% trypsin-EDTA to adjust the cell density to 2 × 10⁻⁶ cells / year. 5 Cells / mL were seeded into 96-well plates, with 100 μL of cell suspension added to each well.

[0107] (5) Drug treatment and inflammation induction Blank control group: 100 μL of cell suspension and 100 μL of culture medium were added to each well, without adding LPS or sample solution.

[0108] LPS group: 100 μL of cell suspension was added to each well, followed by 50 μL of culture medium and then 50 μL of 1 μg / mL LPS solution to induce an inflammatory response.

[0109] Sample group: Add 100 μL of cell suspension to each well, then add 50 μg / mL of sample solution (per well), and pre-treat the 96-well plate in an incubator for 1 hour. After pre-treatment, add 50 μL of 1 μg / mL LPS solution to each well.

[0110] Positive control group: 100 μL of cell suspension was added to each well, followed by 50 μL of 1 μM dexamethasone solution. After pretreatment for 1 hour, 50 μL of 1 μg / mL LPS solution was added.

[0111] Following the group settings, accurately add the corresponding drugs and LPS solution using a multichannel pipette to achieve a final volume of 200 μL per well. When adding the solution, avoid direct impact on the cells; instead, gently add the solution by placing the pipette tip against the edge of the well. After adding the solution, gently shake the 96-well plate to ensure thorough mixing. Seal the 96-well plate with sealing film to prevent moisture evaporation and contamination. Incubate the treated cells in a 37°C, 5% CO2 incubator for 24 hours. During incubation, avoid frequently opening the incubator door to maintain a stable culture environment.

[0112] (6) Detection of inflammatory factors and calculation of results After 24 hours of incubation, remove the 96-well plate from the incubator and place it on an ice pack to prevent degradation of inflammatory factors. Transfer the cell culture supernatant from the 96-well plate to a new 96-well plate, avoiding aspirating cell pellets as much as possible. If further processing of the supernatant is required (e.g., centrifugation to remove cell debris), centrifuge at 1000 rpm for 5 minutes at 4°C and collect the supernatant for detection. Follow the ELISA kit instructions, placing the ELISA plate in a microplate reader. Select the corresponding detection wavelength based on the TNF-α and IL-1α inflammatory factors in the kit and measure the absorbance (OD value) of each well. Plot the standard concentration as the X-axis and the OD value as the Y-axis using four-parameter logistic regression (4-PL) or linear regression to fit the curve. Substitute the sample OD values ​​into the standard curve equation to calculate the TNF-α or IL-1α concentration (pg / mL), as shown in the following formula: TNF-α inhibition rate (%) = (TNF-α concentration in 1 LPS group / TNF-α concentration in the test sample group) × 100%; IL-1α inhibition rate (%) = (IL-1α concentration in 1 LPS group / IL-1α concentration in the test sample group) × 100%; The results are shown in Table 5.

[0113] 2. Tyrosinase activity inhibition test (1) Experimental materials and reagents Reagents: Tyrosinase (recombinant human source, concentration 0.05 mg / mL); L-tyrosine (substrate, concentration 0.15 mmol / L); phosphate buffer (PBS, pH 6.8); test sample (solute consisting of the composition and capsule wall composition, solvent DMSO, concentration 5 mg / mL); positive control: kojic acid (IC50 reference value: 0.005-0.015 mg / mL).

[0114] Instruments: UV spectrophotometer; constant temperature water bath (37℃); cuvette (3mL); pipette (0.1-1mL).

[0115] (2) Experimental Grouping Blank control group (A1): PBS + L-tyrosine + tyrosinase.

[0116] Blank baseline group (A2): PBS + L-tyrosine.

[0117] Positive control group (PC): PBS + L-tyrosine + tyrosinase + kojic acid (final concentration 0.01 mg / mL).

[0118] Sample group (S1-Sn): PBS + L-tyrosine + tyrosinase + test sample solution (5mg / mL).

[0119] (3) Experimental steps Solution preparation: L-Tyrosine solution: Weigh 0.0272 g L-tyrosine, dissolve it in PBS and bring the volume to 100 mL (0.15 mmol / L).

[0120] Tyrosinase solution: Weigh 0.0010 g of tyrosinase, dissolve it in PBS and bring the volume to 20 mL (0.05 mg / mL), and store at -4 °C.

[0121] Reaction conditions: After mixing all test tubes, incubate at 37°C for 30 minutes, add 0.5 mL of tyrosinase solution, mix quickly, and transfer to a cuvette.

[0122] Absorbance measurement: The absorbance (OD value) of each group was measured at a wavelength of 475 nm and recorded as A1, A2, PC, S1-Sn.

[0123] (4) Inhibition rate calculation Inhibition rate (%) = [1(Sn A2) / (A1A2)] × 100% Wherein: Sn: absorbance of sample group / positive control group; A1: absorbance of blank control group; A2: absorbance of blank baseline group; the results are shown in Table 5.

[0124] 3. In vitro DPPH free radical scavenging test (1) Experimental materials and reagents: DPPH free radical reagent: purity ≥98%, stored in the dark; solvent: anhydrous ethanol (analytical grade); samples: the compositions of Examples 1-9 and Comparative Examples 1-8 and the capsule wall composition; positive control sample: vitamin C.

[0125] Instruments: UV-Vis spectrophotometer, analytical balance, pipette (100-1000μL), centrifuge, ultrasonic cleaner, volumetric flasks (50mL, 100mL), test tubes or centrifuge tubes.

[0126] (2) Experimental steps Preparation of DPPH solution: Weigh 4 mg of DPPH powder, dissolve in anhydrous ethanol, and bring the volume to 100 mL. Final concentration: 0.1 mM (approximately 4 mg / L) or adjust to absorbance A0 = 0.6-1.0 (517 nm) based on preliminary experiments. Sonicate for 5 minutes to aid dissolution and store protected from light (use within 3.5 hours).

[0127] Sample / control sample solution preparation: Weigh the sample, dissolve it in DMSO and make up to volume to a final concentration of 5 mg / mL.

[0128] (3) Experimental Groups: Blank control group (A0): 2 mL DPPH solution + 1 mL solvent.

[0129] Sample group (A1): 2 mL DPPH solution + 1 mL sample solution.

[0130] Control sample group (A3): 2 mL DPPH solution + 1 mL control sample solution.

[0131] Sample matrix group (A2): 2 mL solvent + 1 mL sample solution (to exclude interference from the sample's own color).

[0132] Reaction: After mixing each group thoroughly, let it stand in the dark for 30 minutes.

[0133] Centrifugation: Centrifuge at 5000 r / min for 10 minutes (to remove precipitate and avoid interfering with absorbance measurement).

[0134] Absorbance measurement: Take the supernatant and measure the absorbance at a wavelength of 517 nm.

[0135] (4) Parallel experiments and replication Three parallel tubes were set up for each concentration, and A0 (blank control) was remeasured after each concentration was measured.

[0136] (5) Data calculation and result analysis The formula for DPPH free radical scavenging rate is: Scavenging rate (%) = [1(A1 / A3A2) / A0] × 100%; the results are shown in Table 5.

[0137] Table 5 As can be seen from Table 5, when the technical solution provided by the present invention is adopted, the resulting composition has excellent tyrosinase inhibition effect, TNF-α and IL-1α inhibition effect, and can also effectively scavenge DPPH; specifically, the inhibition rate of TNF-α and IL-1α is above 52% and above 51%, respectively, the inhibition rate of tyrosinase is above 50%, and the scavenging rate of DPPH is above 55%. As can be seen from Examples 1-5, the mass fraction of the components in the composition affects the overall performance of the composition. When the mass fraction of the components is further selected within the preferred range, the resulting composition has an inhibition rate of 54% or more against TNF-α and 56% or more against IL-1α, an inhibition rate of 59% or more against tyrosinase, and a scavenging rate of DPPH or more. As can be seen from Examples 1 and 6, and Examples 1 and 7-9, the type of fibronectin and the choice of carrot seed oil also affect the overall performance of the composition; As can be seen from Examples 1 and Comparative Examples 1-4, the overall performance of the resulting composition is significantly reduced when any component in the composition is missing; as can be seen from Examples 1 and Comparative Examples 5-8, the resulting composition cannot achieve the effects of the present invention when other similar components are substituted for the components in the composition. In addition, the experimental data also show that the capsule wall composition has almost no inhibitory effect on TNF-α, IL-1α and tyrosinase, and shows almost no clearance effect on DPPH.

[0138] Example of effect 4 The skincare gel prepared according to the efficacy examples and comparative application examples of this invention underwent mildness testing, including the following steps: Thirty volunteers were recruited, 15 men and 15 women, aged 20-50 years. A closed patch test method was used. Equal volumes (0.020 mL-0.025 mL) of the test sample were placed in a specific patch applicator, which was then applied to the volunteers' arms with hypoallergenic adhesive tape. The patch was gently pressed to ensure even application to the skin and left for 24 hours. The blank control group consisted of distilled water. After 24 hours, the patch applicator was removed, and skin reactions were observed and recorded at 0.5 hours, 24 hours, and 48 hours. The severity of adverse skin reactions is shown in Table 6 below. Table 6 After testing, the skin care gels provided in the application examples and comparative application examples of this invention showed negative reactions after human patch testing, indicating that they are safe and non-irritating to human skin.

[0139] Example 5 The skincare gels prepared in the examples and comparative application examples of this invention have human efficacy in humans. However, because the nanovesicles obtained in comparative application examples 9-11 have low encapsulation rates and poor redox responsiveness, they are not tested for human efficacy. The specific tests include the following steps: 125 Asian adult participants aged 28-40 with sensitive skin and uniform facial pigmentation (such as melasma and sunspots) were randomly divided into 25 groups of 5 participants each. Volunteers applied 1 mL of the sample (prepared from Application Examples 1-16, Control Application Examples 1-10, and the blank application example) to their face twice daily, morning and evening. Data were collected on day 0 (T0), day 14 (T14), and day 28 (T28). After arriving, volunteers washed their faces with facial cleanser and sat quietly for 2 hours in an air-conditioned room with a temperature of 21±1℃ and humidity of 50±10%. Facial imaging was performed using VISIA-CR, blood flow (reflecting inflammatory congestion) was measured using laser Doppler ultrasound, melanin content was measured using a Mexameter, and wrinkle area values ​​(crow's feet, under-eye wrinkles, and nasolabial folds) were determined using PRIMOS-CR. The improvement rate was then calculated as follows: Improvement rate = 1 - (T) 14 / 28 / T0)×100%; the results are shown in Table 7-8; Table 7 Table 8 As can be seen from Tables 7-8, when the technical solution provided by this invention is adopted, the resulting skin care gel has excellent whitening, anti-inflammatory, and antioxidant effects. Specifically, when the skin care gel is prepared using a nanovesicle structure, the improvement rates of melanin content after 14 days and 28 days are above 28% and 37%, respectively; the improvement rates of inflammation after 14 days and 28 days are above 30% and 40%, respectively; the improvement rates of wrinkle area at the corners of the eyes after 14 days and 28 days are above 22% and 31%, respectively; the improvement rates of wrinkle area under the eyes after 14 days and 28 days are above 20% and 28%, respectively; and the improvement rates of wrinkle area at the nasolabial folds after 14 days and 28 days are above 27% and 36%, respectively. As can be seen from Application Examples 1-5, the mass fraction of the components in the composition of the nanovesicles added to the skin care gel affects the overall performance of the composition. When the mass fraction of the components in the composition is further selected within the preferred range, the overall performance of the obtained skin care gel is even better. As can be seen from Application Examples 1 and 6, and Application Examples 1 and 7-9, the type of fibronectin in the nanovesicle composition and the choice of carrot seed oil also affect the overall performance of the skin care gel. As can be seen from Application Example 1 and Comparative Application Examples 1-4, when the composition in the nanovesicles lacks any component, the overall effect of the obtained skin care gel is significantly reduced; as can be seen from Application Example 1 and Comparative Application Examples 5-8, when the components in the composition in the nanovesicles are replaced with other similar components, the obtained skin care gel cannot achieve the effect of the present invention. As can be seen from Application Examples 1 and 15, as well as Application Examples 14 and 16, the long-term effects of the skin care gel are better when nanovesicles are used. In addition, before-and-after photos of volunteers using the skincare gel prepared in Application Example 1 are shown below. Figure 1-2 As shown, from Figure 1-2 It can be clearly seen that the product prepared using the technical solution of this invention has good effects in reducing wrinkles, improving dullness, and improving inflammation.

[0140] Finally, it should be noted that the above embodiments are used to illustrate the technical solutions of the present invention and not to limit the scope of protection of the present invention. Although the present invention has been described in detail with reference to preferred embodiments, those skilled in the art should understand that modifications or equivalent substitutions can be made to the technical solutions of the present invention without departing from the essence and scope of the technical solutions of the present invention.

Claims

1. A composition for targeting and improving dullness and fine lines on sensitive skin, characterized in that, The composition comprises the following components in parts by weight: Oxidized resveratrol 0.01-10 parts, fibronectin 0.01-2 parts, carnosine 0.01-2 parts, carrot seed oil 0.01-3 parts.

2. The composition according to claim 1, characterized in that, The composition comprises the following components in parts by weight: 0.1-5 parts of oxidized resveratrol, 0.01-1 parts of fibronectin, 0.05-1 parts of carnosine, and 0.1-2 parts of carrot seed oil.

3. The composition according to claim 2, characterized in that, The composition comprises the following components in parts by weight: 0.5-1 parts of oxidized resveratrol, 0.3-0.5 parts of fibronectin, 0.1-0.2 parts of carnosine, and 0.5-1 parts of carrot seed oil.

4. The composition according to claim 1, characterized in that, The composition contains 8-38% carrot seed oil by mass.

5. The composition according to claim 1, characterized in that, The carrot seed oil has an acid value ≤3.0mg KOH / g and a saponification value of 3-10mg KOH / g.

6. The composition according to claim 5, characterized in that, The method for preparing carrot seed oil includes the following steps: after drying, carrot seeds are crushed and sieved, then steam distilled, the condensed oil phase is collected and distilled under reduced pressure to obtain carrot seed oil.

7. A nanovesicle that targets and improves dullness and fine lines on sensitive skin, characterized in that, The nanovesicles include a core and a wall, the core comprising the composition as described in any one of claims 1-6, and the wall comprising Tween-80, propylene glycol, and polyethylene glycol-polypropylene sulfide. The mass ratio of Tween-80, propylene glycol, and polyethylene glycol-polypropylene sulfide is (0.01-5):(1-15):(0.01-5).

8. The nanovesicles according to claim 7, characterized in that, The mass ratio of Tween-80, propylene glycol, and polyethylene glycol-polypropylene sulfide is (0.1-1):(3-10):(0.1-1); And / or, the block ratio of polyethylene glycol to polypropylene sulfide in the polyethylene glycol-polypropylene sulfide is (1-2):1; And / or, in the nanovesicles, the mass percentage of the core is 20-36%; And / or, the average particle size of the nanovesicles is 150-300 nm.

9. The use of the composition according to any one of claims 1-6 or the nanovesicles according to any one of claims 7-8 in the preparation of cosmetics.

10. A skin care gel, characterized in that, The skin care gel comprises the composition as described in any one of claims 1-6 or the nanovesicles as described in any one of claims 7-8.

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