A composition for lightening and firming skin, and a method for preparing and using the same

Abstract

This application relates to the field of cosmetics, disclosing a composition for reducing wrinkles and firming the skin, its preparation method, and its application. The composition comprises cosmetically acceptable excipients and the following active ingredients in weight percentages: 0.15%–3.0% palmitoyl pentapeptide-4 encapsulated in flexible nanoliposomes; 0.6%–8.0% a nerve-blocking composition containing acetyl hexapeptide-8 and anti-photoaging plant active ingredients; 1.5%–15.0% a film-forming composition containing enzymatically hydrolyzed peach resin extract, recombinant collagen peptides, and a hyaluronic acid cross-linked polymer; and 0.7%–5.0% a microenvironment synergistic composition containing sea buckthorn extract, tetrahydromethylpyrimidine carboxylic acid, acetyl tetrapeptide-5, and carnosine. This invention overcomes the transdermal barrier of peptides, achieving a highly effective anti-aging effect of reducing wrinkles and firming the skin.

Description

Technical Field

[0001] This application relates to the field of cosmetic technology, and more specifically, to a composition for reducing wrinkles and firming the skin, a method for preparing the composition, and its application. Background Technology

[0002] The skin around the eyes and face, especially the periocular area, is the thinnest and most vascularized skin tissue in the human body. It is highly susceptible to dynamic wrinkles and static sagging due to aging, frequent muscle contractions (expression lines), UV radiation, and collagen loss. Currently, the cosmetic industry widely uses peptides, collagen, and plant extracts as core active ingredients in anti-aging compositions. These ingredients have demonstrated good potential in promoting extracellular matrix production and reducing wrinkles, both theoretically and in vitro.

[0003] However, existing anti-aging compositions face insurmountable technical bottlenecks in practical applications. Firstly, their transdermal penetration efficiency is extremely low: the molecular weight of most anti-wrinkle peptides is between 800 and 1600 Da, far exceeding the 500 Da absorption limit of the human stratum corneum. When added to traditional formulations in a free state, over 90% of the peptides remain only on the skin surface, failing to reach the fibroblast target in the dermis, resulting in actual wrinkle-reducing effects far below theoretical expectations. Secondly, there are incompatibilities in the formulation system and the easy deactivation of active ingredients: when attempting to combine natural macromolecular film-forming agents (such as traditional peach gum / peach resin crude extract) with peptides, existing technologies easily lead to charge complexation, causing peptide precipitation or severe pilling during product application; simultaneously, conventional high-temperature emulsification processes and acidic plant extracts (such as sea buckthorn extract rich in vitamin C) rapidly destroy the chemical stability of the peptides. Finally, there is a lack of synergistic mechanisms in the cellular microenvironment: existing products often provide collagen synthesis peptides or collagen in isolation, but ignore the fact that aging cells lack the necessary hydroxylation coenzymes and protective mechanisms to cope with changes in physical osmotic pressure when executing synthesis instructions, resulting in low synthesis efficiency. Summary of the Invention

[0004] To address the aforementioned issues, this application provides a composition for reducing wrinkles and firming the skin, along with its preparation method and application.

[0005] The first part of this application provides a composition for reducing wrinkles and firming the skin, which adopts the following technical solution: A composition for reducing wrinkles and firming the skin, comprising, by weight percentage, the following active ingredients: Component A: 0.15% to 3.0% of dermal layer-targeting peptides, wherein the dermal layer-targeting peptides include palmitoyl pentapeptide-4 encapsulated in flexible nanoliposomes; Component B: 0.6% to 8.0% of a nerve-blocking composition, wherein the nerve-blocking composition comprises acetyl hexapeptide-8 and anti-photoaging plant active ingredients; Component C: 1.5% to 15.0% of an interpenetrating network film-forming composition, wherein the interpenetrating network film-forming composition comprises enzymatically hydrolyzed peach resin extract, recombinant collagen peptides, and hyaluronic acid crosslinked polymer; Component D: 0.7%–5.0% of a mitochondrial-microenvironment synergistic composition comprising sea buckthorn extract, tetrahydromethylpyrimidine carboxylic acid, acetyl tetrapeptide-5, and carnosine.

[0006] By employing the above technical solutions, component A overcomes the transdermal barrier of large-molecule peptides through flexible nanoliposomes, delivering deep anti-aging signals to the dermis to activate collagen regeneration; component B remains in a free state at the epidermal-dermal junction, instantly blocking nerve transmission in facial muscles and smoothing dynamic wrinkles; component C forms an interpenetrating polymer network (IPN) on the skin surface, providing not only an immediate physical stretching and wrinkle-smoothing effect, but also generating downward osmotic pressure through physical contraction, forcibly driving components A and B to penetrate downwards; component D provides essential hydroxylation coenzymes and osmotic pressure protection for collagen synthesis in basal fibroblasts. Furthermore, the sea buckthorn extract and carnosine in component D, while empowering collagen synthesis, possess potent antioxidant and anti-glycation properties that can scavenge excess free radicals (ROS) and advanced glycation end products (AGEs), thereby helping to improve yellowing and dullness of the skin around the eyes to some extent.

[0007] Optionally, the membrane material of the flexible nanoliposomes is composed of a base phospholipid and an edge activator; the base phospholipid is selected from soybean lecithin or hydrogenated lecithin, and the edge activator is selected from at least one of sodium deoxycholate, Tween-80 or dipotassium glycyrrhizate; the mass ratio of the base phospholipid to the edge activator is 3:1 to 10:1; the average particle size of the flexible nanoliposomes is distributed between 50 nm and 120 nm.

[0008] By employing the above-mentioned technical solution, conventional rigid liposomes are prone to rupture when encountering intercellular spaces smaller than themselves. However, this application introduces a specific proportion of edge activator, significantly increasing the flexibility of the liposome bilayer. Driven by the hydration gradient difference formed by the evaporation of water from the skin surface, the liposome can undergo reversible elastic deformation, passing through intercellular lipid channels, resulting in an order-of-magnitude increase in the deep delivery of peptides. If dipotassium glycyrrhizate is selected as the edge activator, it not only exerts a softening effect but also possesses soothing properties, which can fine-tune periorbital microcirculation inflammation and help reduce localized darkening caused by post-inflammatory hyperpigmentation.

[0009] Optionally, the enzymatically hydrolyzed peach resin extract is an oligosaccharide fragment with a molecular weight distribution of 5000 Da to 10000 Da; the enzymatically hydrolyzed peach resin extract, recombinant collagen peptide, and hyaluronic acid crosslinking polymer are crosslinked by non-covalent bonds.

[0010] By employing the above technical solution, oligosaccharide fragments with a cutoff of 5000Da to 10000Da exhibit excellent compatibility and do not undergo charge precipitation with peptides. These fragments are non-covalently cross-linked with recombinant collagen peptides and hyaluronic acid polymers to form a microporous, breathable interpenetrating network (IPN) film. The directional tension generated during drying and shrinkage of this film not only supports the sagging eye contour but also generates inward mechanical pressure, significantly improving the transdermal absorption efficiency of the underlying active substances.

[0011] Optionally, the mitochondrial and microenvironment synergistic composition contains, by weight percentage, 0.5% to 3.0% of sea buckthorn extract, 0.1% to 1.0% of tetrahydromethylpyrimidine carboxylic acid, 0.05% to 0.5% of acetyl tetrapeptide-5, and 0.05% to 0.5% of carnosine.

[0012] By employing the above technical solution, when physical osmotic pressure is applied to the IPN membrane, tetrahydromethylpyrimidine carboxylic acid acts as a polar osmotic pressure compensator, protecting the three-dimensional structure of cells from mechanical stress damage. Simultaneously, sea buckthorn extract provides essential hydroxylated coenzymes (natural vitamin C) for the transformation of procollagen into a mature collagen network. The highly active vitamin C, in synergy with the anti-glycation agent carnosine and the anti-inflammatory agent acetyl tetrapeptide-5, effectively accelerates the metabolism of accumulated lipofuscin and melanin around the eyes, thereby achieving secondary cosmetic effects such as brightening the skin tone around the eyes and reducing dark circles.

[0013] Optionally, the cosmetic excipients include a moisturizing matrix composed of sodium hyaluronates of different molecular weights, wherein the molecular weight gradient of sodium hyaluronates in the moisturizing matrix covers 10 kDa to 2000 kDa.

[0014] By employing the above-mentioned technical solution, a gradient distribution of 10kDa to 2000kDa ensures ample moisture from the stratum corneum to the dermis. Large molecules form a water-locking barrier in the epidermis, while small molecules penetrate deep into the dermis, causing the extracellular matrix (ECM) to swell, providing an ideal aqueous channel for the downward diffusion of flexible liposomes. The high hydration state of the dermis significantly enhances the skin's optical refractive index, making the skin appear fuller and brighter, further reducing the appearance of dark circles around the eyes.

[0015] Optionally, the preparation method of the dermal layer-targeting peptide includes the following steps: The basic phospholipids and edge activators were dissolved in anhydrous ethanol, and the organic solvent was removed by rotary evaporation under reduced pressure to form a uniform lipid film on the inner wall of the container. The palmitoyl pentapeptide-4 was dissolved in a phosphate buffer solution at pH 5.5 to obtain an aqueous phase containing the polypeptide. An aqueous phase containing peptides was added to a lipid membrane and hydrated at a low temperature of 20℃ to 30℃ to obtain a primary peptide liposome suspension. The primary polypeptide liposome suspension was placed in a high-pressure microfluidic homogenizer and homogenized for 3 to 5 cycles at a low-temperature cooling cycle with an operating pressure of 500 bar to 1000 bar. Then, the granules were extruded through a 100 nm polycarbonate microporous membrane to obtain flexible nanoliposomes encapsulating the dipeptides.

[0016] By employing the above-mentioned technical solution, the peptides, which are extremely sensitive to high temperatures and shear forces, are addressed through a process that utilizes low-temperature hydration at 20℃–30℃, supplemented by high-pressure microfluidic homogenization under low-temperature cooling cycles. This avoids peptide denaturation caused by conventional high-temperature emulsification. Operating pressures of 500 bar–1000 bar and extrusion through specific microporous membranes ensure the uniformity of flexible vesicle particle size, thereby guaranteeing the consistency and stability of the transdermal efficacy rate of anti-aging products in practical applications.

[0017] Optionally, the method for preparing the enzymatically hydrolyzed peach resin extract includes the following steps: Natural peach resin was placed in deionized water to swell and heated to 80℃~90℃ to fully dissolve it. After filtering to remove impurities, a crude peach resin polysaccharide solution was obtained. The system temperature was lowered to 45℃~50℃, the pH was adjusted to 4.5~5.5 using citrate buffer, a complex glycosidase composed of cellulase and pectinase was added, and the system was stirred at a constant temperature for 2~4 hours to carry out the directional enzymatic hydrolysis reaction. The enzyme is inactivated by heating to 80℃~85℃, and then the hydrolysate is cooled and passed through ultrafiltration membranes with molecular weight cutoffs of 10000Da and 5000Da in sequence. The permeate with molecular weight cutoffs between 5000Da and 10000Da is collected, concentrated and dried to obtain the enzymatically hydrolyzed peach resin extract.

[0018] By employing the above-mentioned technical solution, unlike highly destructive acid-base hydrolysis, this application uses a complex glycosidase to perform gentle, directional cleavage at specific pH and temperature, preserving the film-forming active groups of the polysaccharide side chains. Combined with the cascade retention of the ultrafiltration membrane, it completely eliminates ultra-large molecules that cause skin acne and miscellaneous sugars smaller than 5kDa that lack physical stretching force, providing the product with high-purity, high-stability key film-forming framework raw materials.

[0019] Part Two, this application provides a method for using a composition to reduce wrinkles and firm the skin, characterized by comprising the following steps: Step S1: Add the enzymatically hydrolyzed peach resin extract, recombinant collagen peptides, hyaluronic acid cross-linked polymers and cosmetically acceptable excipients to deionized water, heat to 75℃~85℃ for homogenization, and then cool to below 40℃ to form a hydrogel matrix. Step S2: Adjust the pH of the hydrogel matrix to 5.0-5.5, add the mitochondrial and microenvironment synergistic composition, and stir to evenly disperse it in the hydrogel matrix; Step S3: Under the condition of temperature control at 25℃~30℃, add the dermal layer targeting peptide and the nerve blocking composition, stir evenly with a shear rate of ≤1500rpm, and let stand to defoam to obtain the composition for reducing wrinkles and firming.

[0020] By employing the above technical solution, the macromolecular backbone is first hydrated at high temperature; after cooling, the pH of the system is precisely adjusted to the weakly acidic range (5.0–5.5) where the peptides are most stable, and then a coenzyme matrix is ​​added; finally, the most fragile liposomal peptides and neuropeptides are cold-mixed under room temperature and low shear force conditions. This process logic avoids peptide inactivation caused by physical shear force demulsification and pH imbalance, ensuring the long-lasting activity of the anti-aging product during its shelf life.

[0021] Part Three, this application provides the use of a composition for reducing wrinkles and firming the skin in the preparation of a non-therapeutic cosmetic for reducing wrinkles and improving skin firmness on the face or around the eyes.

[0022] In summary, this application has the following beneficial effects: 1. This application employs a three-dimensional, layered anti-aging system constructed from dermal-targeting peptides, free-state neuroprotective peptides, interpenetrating network film-forming compositions, and a microenvironment synergistic composition encapsulated in flexible nanoliposomes. This system not only forces deep penetration of peptides through inward contraction and compression of the physical film, but also provides essential hydroxylation coenzymes and osmotic pressure protection for collagen synthesis in the basal fibroblasts. From targeted signal delivery and physical compression-promoted penetration to cellular microenvironment empowerment, this system significantly enhances the transdermal absorption rate of peptides and the efficiency of collagen regeneration, achieving a highly effective core anti-aging effect of reducing wrinkles and firming the skin. Simultaneously, the highly active coenzymes and anti-oxidative and anti-glycation components in the microenvironment composition work synergistically to effectively accelerate the metabolism of accumulated pigments and lipofuscin around the eyes, helping to improve dark circles under the eyes.

[0023] 2. This application preferably uses flexible nanoliposomes modified with a specific edge activator and enzymatically hydrolyzed peach resin extract. The flexible nanoliposomes possess the ability to undergo reversible elastic deformation driven by a hydration gradient difference, enabling them to smoothly penetrate the narrow intercellular spaces of the stratum corneum and overcome the transdermal barrier of traditional large-molecule peptides. The enzymatically hydrolyzed peach resin extract not only solves the industry pain point of traditional natural polysaccharides easily forming charge complexes with peptides, leading to "pilling," but also crosslinks with recombinant collagen peptides to form a microporous, breathable interpenetrating polymer network (IPN), providing an immediate physical stretching and smoothing effect as well as long-lasting aqueous penetration channels.

[0024] 3. The method of this application, through a segmented preparation process, effectively overcomes the formulation challenges of antagonism and degradation among active components in complex anti-aging compositions. This method involves first hydrating the macromolecular backbone at high temperature, then precisely adjusting the pH of the system to the weakly acidic range (5.0–5.5) where the peptides are most stable, and introducing a coenzyme matrix. Finally, under room temperature and low shear stress conditions, liposomal peptides and neuropeptides, which are extremely sensitive to heat and physical effects, are cold-mixed in. This avoids peptide inactivation caused by high-temperature denaturation, pH imbalance, and strong shear demulsification, maximizing the activity and stability of the anti-aging and firming product during long-term storage and practical application. Attached Figure Description

[0025] Figure 1 The image shows a comparison of eye wrinkles of the first volunteer randomly selected in Example 1 of the present invention before (day 0) and after (day 28) continuous use of the composition. Figure 2 The image shows a comparison of eye wrinkles of a second volunteer randomly selected in Example 1 of the present invention before (day 0) and after (day 28) continuous use of the composition. Figure 3 The image shows a comparison of eye wrinkles of a third volunteer randomly selected in Example 1 of the present invention before (day 0) and after (day 28) continuous use of the composition. Figure 4 The image shows a comparison of eye wrinkles of a fourth volunteer randomly selected in Example 1 of the present invention before (day 0) and after (day 28) continuous use of the composition. Figure 5 This is a comparison image of the depth of eye wrinkles of a volunteer randomly selected in Example 1 of the present invention before (day 0) and after (day 28) continuous use of the composition. Detailed Implementation

[0026] The following detailed description of this application is provided in conjunction with the embodiments. It should be noted that: unless otherwise specified, the conditions in the following embodiments are performed under conventional conditions or conditions recommended by the manufacturer. Unless otherwise specified, the raw materials used in the following embodiments are all from commercially available sources.

[0027] Example 1: This example provides a composition for reducing wrinkles and firming the skin, with the following mass percentage formulation of each component: Component A (dermal targeting peptide, total 1.5%), palmitoyl pentapeptide-4 (Dongguan Weiqi Technology Co., Ltd., WKPep®Pal-KTTKS) 0.5%, flexible nanoliposome membrane material (soybean lecithin and sodium deoxycholate, mass ratio 6:1) 1%.

[0028] Component B (nerve blocking composition, total 4.0%): Acetyl hexapeptide-8 (Dongguan Weiqi Technology Co., Ltd., WKPep®Eyepep02 Eye Peptide 02) 2.0%, Anti-photoaging plant active ingredient (rose extract, commercially available) 2.0%.

[0029] Component C (interpenetrating network film-forming composition, total 8.0%): enzymatic hydrolysis of peach resin extract (manufacturer: Shanghai Gakai Biotechnology Co., Ltd., trade name: ImmedelinePG) 4%, recombinant collagen peptide (Zhejiang Zhuji Juyuan Biotechnology Co., Ltd., Beikeli® 6D recombinant collagen (MC02)) 2%, hyaluronic acid cross-linked polymer (Bloomage Biotechnology Co., Ltd., HymagicTM-4D4D hyaluronic acid) 2%.

[0030] Component D (Mitochondrial and Microenvironment Synergistic Composition, Total 2.5%): The mitochondrial and microenvironment composition includes 1.5% sea buckthorn extract (Zhejiang Zhongke Zhongzhi Biotechnology Co., Ltd., Tangola™-OPC sea buckthorn proanthocyanidins), 0.5% tetrahydromethylpyrimidine carboxylic acid (Shandong Juntian Pharmaceutical Co., Ltd., KTcos™ ectoine), 0.2% acetyl tetrapeptide-5 (Dongguan Weiqi Technology Co., Ltd., WKPep®Eyepep02), 0.3% carnosine (Dongguan Weiqi Technology Co., Ltd., WKPep® Carnosine), with the balance being deionized water.

[0031] The remainder consists of cosmetically acceptable excipients: 5.0% moisturizing matrix (containing a compound of 10kDa, 500kDa and 2000kDa sodium hyaluronate), 0.5% phenoxyethanol, and the remainder is deionized water.

[0032] The preparation method includes the following stages: Phase 1: Preparation of enzymatic hydrolysis of peach resin extract (1) Natural peach resin was placed in deionized water to swell, heated to 85°C to fully dissolve it, and filtered to remove impurities to obtain a crude peach resin polysaccharide solution. (2) Reduce the system temperature to 48°C, adjust the pH of the system to 5.0 using citrate buffer, add a complex glycosidase composed of cellulase and pectinase, and carry out a directional enzymatic hydrolysis reaction for 3 hours with constant temperature stirring. (3) Heat to 82°C and keep warm for 15 minutes to inactivate the enzyme. After cooling, pass through ultrafiltration membrane modules with molecular weight cutoffs of 10000 Da and 5000 Da in sequence. Collect the permeate section with molecular weight cutoffs between 5000 Da and 10000 Da. After concentration and drying, the desired enzymatically hydrolyzed peach resin extract is obtained.

[0033] Phase Two: Preparation of Dermal Targeting Peptides (1) Soybean lecithin and sodium deoxycholate (mass ratio 6:1) were dissolved in anhydrous ethanol, and the organic solvent was removed by rotary evaporation under reduced pressure to form a uniform lipid film on the inner wall of a round-bottom flask. (2) Dissolve palmitoyl pentapeptide-4 in phosphate buffer at pH 5.5 to obtain an aqueous phase containing the polypeptide; (3) Add the aqueous phase containing the polypeptide to the above lipid film and perform hydration shaking at a low temperature of 25°C to obtain a primary polypeptide liposome suspension. (4) The primary suspension was placed in a high-pressure microfluidic homogenizer. Under the protection of the low-temperature cooling circulation system, the operating pressure was set to 800 bar for four cycles of homogenization. Then, the granules were extruded through a 100 nm polycarbonate microporous filter membrane to obtain flexible nanoliposomes (component A) with an average particle size of about 80 nm that encapsulate the dipeptide.

[0034] Phase 3: Preparation of the final product of the wrinkle-reducing and firming composition Step S1: Add component C (enzymatically hydrolyzed peach resin extract, recombinant collagen peptide, hyaluronic acid cross-linked polymer) and moisturizing matrix to deionized water, heat to 80°C and stir homogenously for 30 minutes to allow it to fully expand and hydrate, then gradually cool to 38°C to form a hydrogel matrix with a microporous structure. Step S2: Precisely adjust the pH of the hydrogel matrix to 5.2, slowly add component D (mitochondrial and microenvironment synergistic composition), and stir at medium speed to evenly disperse it in the grid of the hydrogel matrix; Step S3: Under the condition of temperature control at 28°C, add the above-prepared component A (targeting peptide composition), component B (nerve blocking composition) and preservative in sequence, stir evenly with a low shear rate of 1000 rpm, and let stand for vacuum degassing to obtain the composition for reducing wrinkles and firming.

[0035] Example 2: The only difference between this example and Example 1 is the amount of each active ingredient added. All parameters are taken as the lower limit of the scope of protection of the claims. The specific formula is as follows: Component A: 0.15%; Component B: 0.6%; Component C: 1.5%; Component D: 0.7% (including 0.5% sea buckthorn extract, 0.1% tetrahydromethylpyrimidine carboxylic acid, 0.05% acetyl tetrapeptide-5, and 0.05% carnosine). The balance is deionized water and auxiliary matrix. The preparation process is exactly the same as in Example 1.

[0036] Example 3: The only difference between this example and Example 1 is the amount of each active ingredient added. All parameters are taken as the upper limit of the scope of protection of the claims. The specific formula is as follows: Component A: 3.0%; Component B: 8.0%; Component C: 15.0%; Component D: 5.0% (including 3.0% sea buckthorn extract, 1.0% tetrahydromethylpyrimidine carboxylic acid, 0.5% acetyl tetrapeptide-5, and 0.5% carnosine). The balance is deionized water and auxiliary matrix. The preparation process is exactly the same as in Example 1.

[0037] Example 4: The only difference between this example and Example 1 is in stage two (preparation of dermal-targeting peptides): the membrane material for the flexible nanoliposomes uses hydrogenated lecithin and Tween-80 in a mass ratio of 3:1; the high-pressure microfluidic operation pressure is adjusted to 1000 bar, and the average particle size of the obtained flexible nanoliposomes is approximately 50 nm. The remaining formulations and processes are exactly the same as in Example 1.

[0038] Example 5: The only difference between this example and Example 1 is in stage two (preparation of dermal-targeting peptides): the membrane material for the flexible nanoliposomes uses soybean lecithin and dipotassium glycyrrhizate in a mass ratio of 10:1; the high-pressure microfluidic operation pressure is adjusted to 500 bar, and the average particle size of the obtained flexible nanoliposomes is approximately 120 nm. The remaining formulation and process are exactly the same as in Example 1.

[0039] Comparative Example 1 The only difference between this comparative example and Example 1 is that the liposome preparation process in stage two is skipped. That is, the base phospholipids and edge activators are not used; in step S3, the unencapsulated palmitoyl pentapeptide-4 is directly added to the hydrogel matrix. The remaining formulation and process are the same as in Example 1.

[0040] Comparative Example 2 The only difference between this comparative example and Example 1 is that the enzymatic hydrolysis process in stage one is skipped. Specifically, ordinary natural peach resin water extract (without the use of complex glycosidases for cleavage or ultrafiltration membrane molecular weight screening) is used in equal amounts to replace the "enzymatically hydrolyzed peach resin extract" in Example 1. The remaining formulation and process are the same as in Example 1.

[0041] Comparative Example 3 The only difference between this comparative example and Example 1 is that component D (mitochondrial and microenvironment synergistic composition) was completely removed from the formulation, i.e., no sea buckthorn extract, tetrahydromethylpyrimidine carboxylic acid, acetyl tetrapeptide-5, and carnosine were added, and the removed mass fraction was made up by deionized water. The rest of the formulation and process are the same as in Example 1.

[0042] Comparative Example 4 The only difference between this comparative example and Example 1 is that the segmented, pH-adjusted, and cold-mixing anchoring process described in claim 8 is not used. During preparation, components A, B, C, D, and all excipients are added to an emulsifying pot all at once, and emulsified directly at 80°C using a high-shear homogenizer at 8000 rpm for 20 minutes, followed by cooling and discharge. The types and amounts of the formulation components are exactly the same as in Example 1.

[0043] Test methods Volunteer recruitment excluded pregnant or breastfeeding women, those with active facial infections or severe skin diseases, and those who had used glucocorticoids or immunosuppressants within one month prior to the test. Ten female volunteers aged 25-45 were selected for each group. After washing their face with water morning and evening, they applied an eye serum to the skin around their eyes, massaging it in circular motions until fully absorbed, for 28 consecutive days.

[0044] The tests were conducted in a constant temperature and humidity laboratory at 21±1℃ and 50±5%. Tests were performed on day 0 and again from day 28 of continuous sample use, with each test taking place between 9 and 11 AM. The number and depth of wrinkles under the eyes and at the outer corners of the eyes were measured using a VISIA skin image analyzer (model Generation 7, manufactured by Canfield, USA).

[0045] Table 1. VISIA Test Data: Before and After Using Eye Serum (Before Trial VS 28 Days After Trial) Example 1 101 57 44% Example 2 105 76 28% Example 3 98 57 42% Example 4 110 68 38% Example 5 104 67 36% Comparative Example 1 108 92 15% Comparative Example 2 102 80 22% Comparative Example 3 106 78 26% Comparative Example 4 100 88 12% As shown in Table 1, after 28 days of continuous use, the compositions of Examples 1-5 significantly improved the number of wrinkles around the eyes by 28% to 44%, indicating that the formulation composition and parameter range specified in this application can effectively achieve the purpose of reducing wrinkles and firming the skin. Among them, Example 1 (with all parameters at the optimal median value) achieved the best improvement rate of 44%. Examples 4 and 5, by changing the membrane material ratio and particle size of the flexible nanoliposomes, still maintained an excellent improvement rate of over 36%, proving that the flexible nanoliposomes described in this application have extremely strong transdermal absorption promotion capabilities in the 50nm-120nm range.

[0046] Comparative Example 1, which did not use flexible nanoliposome technology and added peptides in a free state, saw its wrinkle improvement rate plummet to 15% (this improvement mainly stemmed from the epidermal hydration and plumping effects of moisturizers such as hyaluronic acid). This clearly demonstrates that conventional large-molecule peptides struggle to penetrate the stratum corneum, while the flexible nanoliposomes used in Component A of this application, due to their reversible deformation capabilities imparted by edge activators, can effectively deliver peptides to the dermis through intercellular spaces, a crucial condition for achieving deep anti-aging.

[0047] Comparative Example 2 used ordinary natural peach resin that had not undergone targeted enzymatic hydrolysis, and its improvement rate was only 22%. This is because ordinary macromolecular polysaccharides not only tend to cause pilling on the skin surface, hindering absorption, but also cannot form a tight interpenetrating microporous network (IPN) with collagen peptides. Example 1 (44%) showed a significantly better improvement rate in eye wrinkles than Comparative Example 2, with the extract obtained through enzymatic hydrolysis to a specific molecular weight (5000-10000 Da).

[0048] Comparative Example 3 removed sea buckthorn extract and ectoine D components from the formula. Although the peptide signal could enter the skin, the improvement rate was only 26% due to the lack of hydroxylated coenzyme (vitamin C) required for collagen synthesis in the basal fibroblasts and the lack of osmotic pressure protection.

[0049] Comparative Example 4, which employed a conventional high-temperature one-pot method and high-shear emulsification process, resulted in the lowest improvement rate (only 12%). This demonstrates that the peptides and nanoliposome vesicles in the composition of this application are extremely sensitive to high temperatures and high shear forces. The "precise pH adjustment after cooling, low-temperature cold preparation, and low-shear anchoring" process specified in the method of this application is a key technical feature that protects this complex active system from damage and ensures the clinical anti-aging efficacy of the final product.

[0050] In the appendix Figure 1-4 It is clearly visible that after using the product of Example 1, wrinkles are significantly reduced and the skin around the eyes is smoother.

[0051] In the appendix Figure 5 In the image, yellow / red represents protrusions, and blue / green represents depressions. More blue indicates a deeper depression and more pronounced wrinkles, while more red indicates shallower wrinkles. It can be seen that in the same location, after application, the area of ​​the blue-colored portion significantly decreases, indicating that the wrinkles are noticeably shallower.

[0052] This specific embodiment is merely an explanation of this application and is not intended to limit it. After reading this specification, those skilled in the art can make modifications to this embodiment without contributing any inventive step, but such modifications are protected by patent law as long as they fall within the scope of the claims of this application.

Claims

1. A composition for reducing wrinkles and firming the skin, characterized in that, Its active ingredients, by weight percentage, include the following components: Component A: 0.15% to 3.0% of dermal layer-targeting peptides, wherein the dermal layer-targeting peptides include palmitoyl pentapeptide-4 encapsulated in flexible nanoliposomes; Component B: 0.6% to 8.0% of a nerve-blocking composition, wherein the nerve-blocking composition comprises acetyl hexapeptide-8 and anti-photoaging plant active ingredients; Component C: 1.5% to 15.0% of an interpenetrating network film-forming composition, wherein the interpenetrating network film-forming composition comprises enzymatically hydrolyzed peach resin extract, recombinant collagen peptides, and hyaluronic acid crosslinked polymer; Component D: 0.7%–5.0% of a mitochondrial-microenvironment synergistic composition comprising sea buckthorn extract, tetrahydromethylpyrimidine carboxylic acid, acetyl tetrapeptide-5, and carnosine.

2. The composition according to claim 1, characterized in that, The membrane material of the flexible nanoliposomes is composed of a base phospholipid and an edge activator; the base phospholipid is selected from soybean lecithin or hydrogenated lecithin, and the edge activator is selected from at least one of sodium deoxycholate, Tween-80 or dipotassium glycyrrhizate; the mass ratio of the base phospholipid to the edge activator is 3:1 to 10:1; the average particle size of the flexible nanoliposomes is distributed between 50 nm and 120 nm.

3. The composition according to claim 1, characterized in that, The enzymatically hydrolyzed peach resin extract is an oligosaccharide fragment with a molecular weight distribution of 5000 Da to 10000 Da; the enzymatically hydrolyzed peach resin extract, recombinant collagen peptide and hyaluronic acid crosslinked polymer are crosslinked by non-covalent bonds.

4. The composition according to claim 1, characterized in that, The mitochondrial and microenvironment synergistic composition contains, by weight percentage, 0.5% to 3.0% sea buckthorn extract, 0.1% to 1.0% tetrahydromethylpyrimidine carboxylic acid, 0.05% to 0.5% acetyl tetrapeptide-5, and 0.05% to 0.5% carnosine.

5. The composition according to claim 1, characterized in that, The composition further includes cosmetically acceptable excipients comprising a moisturizing matrix composed of sodium hyaluronates of varying molecular weights, wherein the molecular weight gradient of the sodium hyaluronates in the moisturizing matrix covers 10 kDa to 2000 kDa.

6. The composition according to claim 1, characterized in that, The preparation method of the dermal layer-targeting peptide includes the following steps: The basic phospholipids and edge activators were dissolved in anhydrous ethanol, and the organic solvent was removed by rotary evaporation under reduced pressure to form a uniform lipid film on the inner wall of the container. Palmitoyl pentapeptide-4 was dissolved in phosphate buffer at pH 5.5 to obtain an aqueous phase containing the peptide. An aqueous phase containing peptides was added to a lipid membrane and hydrated at a low temperature of 20℃ to 30℃ to obtain a primary peptide liposome suspension. The primary polypeptide liposome suspension was placed in a high-pressure microfluidic homogenizer and homogenized for 3 to 5 cycles at a low-temperature cooling cycle with an operating pressure of 500 bar to 1000 bar. Then, it was extruded and granulated through a 100 nm polycarbonate microporous filter membrane.

7. The composition according to claim 1, characterized in that, The preparation method of the enzymatically hydrolyzed peach resin extract includes the following steps: Natural peach resin was placed in deionized water to swell and heated to 80℃~90℃ to fully dissolve it. After filtering to remove impurities, a crude peach resin polysaccharide solution was obtained. The system temperature was lowered to 45℃~50℃, the pH was adjusted to 4.5~5.5 using citrate buffer, a complex glycosidase composed of cellulase and pectinase was added, and the system was stirred at a constant temperature for 2~4 hours to carry out the directional enzymatic hydrolysis reaction. The enzyme is inactivated by heating to 80℃~85℃, and then the hydrolysate is cooled and passed through ultrafiltration membranes with molecular weight cutoffs of 10000Da and 5000Da in sequence. The permeate with molecular weight cutoffs between 5000Da and 10000Da is collected, concentrated and dried to obtain the enzymatically hydrolyzed peach resin extract.

8. A method for preparing the composition for wrinkle reduction and firming as described in any one of claims 1 to 7, characterized in that, Includes the following steps: Step S1: Add the enzymatically hydrolyzed peach resin extract, recombinant collagen peptides, hyaluronic acid cross-linked polymers and cosmetically acceptable excipients to deionized water, heat to 75℃~85℃ for homogenization, and then cool to below 40℃ to form a hydrogel matrix. Step S2: Adjust the pH of the hydrogel matrix to 5.0-5.5, add the mitochondrial and microenvironment synergistic composition, and stir to make it evenly dispersed in the hydrogel matrix; Step S3: Under the condition of temperature control at 25℃~30℃, add the dermal layer targeting peptide and the nerve blocking composition, stir evenly with a shear rate of ≤1500rpm, and let stand to defoam to obtain the composition for reducing wrinkles and firming.

9. The use of a composition for reducing wrinkles and firming as described in any one of claims 1 to 7, or a composition prepared by the method of claim 8, in the preparation of a non-therapeutic cosmetic for reducing wrinkles and improving skin firmness on the face or around the eyes.

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