A whitening and freckle lightening composition containing a polygonum cuspidatum-derived niosome and a preparation method thereof

Abstract

This invention discloses a whitening and spot-fading composition containing Polygonum cuspidatum-derived vesicles and its preparation method, belonging to the field of cosmetic technology. The whitening and spot-fading composition includes Polygonum cuspidatum-derived vesicles, whitening active ingredients, soothing and repairing ingredients, moisturizers, thickeners, antioxidants, preservatives, pH adjusters, and water; wherein, the Polygonum cuspidatum-derived vesicles are added to the skincare product matrix as active delivery and skin conditioning ingredients. This invention also provides a method for preparing the whitening and spot-fading composition, including steps such as aqueous phase preparation, dissolving the whitening active ingredients, adding Polygonum cuspidatum-derived vesicles after cooling, low-shear mixing, pH adjustment, defoaming, filtration, and filling. The composition can be used to prepare serums, lotions, creams, gels, masks, or freeze-dried compositions with whitening, brightening, dullness-improving, and skin barrier-repairing effects.

Description

Technical Field

[0001] This invention belongs to the field of cosmetic technology, and in particular relates to a whitening and spot-fading composition containing Polygonum cuspidatum-derived vesicles and its preparation method. Background Technology

[0002] With the development of the functional skincare market, products with whitening, brightening, dullness-improving, and pigmentation-fading effects have become an important direction in skincare research and development. Skin color and dullness are influenced by multiple factors, including melanin production and transport, stratum corneum metabolism, post-inflammatory hyperpigmentation, oxidative stress levels, and skin barrier function. Currently, whitening skincare products commonly use niacinamide, tranexamic acid, ascorbic acid and its derivatives, alpha-arbutin, and licorice root extract as active ingredients. However, in actual formulation applications, some ingredients may have issues such as insufficient stability, irritation risks, limited compatibility, low transdermal absorption efficiency, or difficulty in simultaneously achieving whitening and barrier repair. Therefore, developing a complex skincare system that combines whitening, soothing, antioxidant, and formulation stability still has practical application value.

[0003] Plant-derived vesicles generally refer to nanoscale membrane-like vesicle structures derived from plant tissues or plant cells. They typically contain lipids, proteins, polysaccharides, small molecule active substances, and nucleic acids, and have certain application potential in the cosmetics and skin care fields. Polygonum cuspidatum, a commonly used plant resource, is known to contain various plant active ingredients such as resveratrol, polygalactosides, and emodin. Existing publicly available technologies have reported applications of Polygonum cuspidatum cell vesicles, Polygonum cuspidatum exosomes, or compound compositions containing Polygonum cuspidatum extracellular vesicles in oral care, soothing skin care, and hair growth and anti-hair loss. However, there is still room for improvement in the technical solutions for using Polygonum cuspidatum-derived vesicles as functional raw materials in skin care compositions, and for combining them with whitening active ingredients, moisturizing ingredients, and soothing and repairing ingredients to construct whitening and spot-fading compositions with whitening effects. Furthermore, as nanoscale membrane raw materials, plant vesicles may be affected by factors such as temperature, shear force, pH, solvent system, preservative system, and thickening system during the formulation and processing of skin care products, thereby affecting their particle size distribution, dispersion state, and formulation stability. Summary of the Invention

[0004] To address the aforementioned technical problems, this invention proposes a whitening and spot-fading composition containing Polygonum cuspidatum-derived vesicles and a preparation method thereof. This allows Polygonum cuspidatum-derived vesicles to be compounded with whitening active ingredients, moisturizers, soothing and repairing ingredients, and stabilizing matrix under relatively mild formulation conditions, thereby improving the compatibility, stability, and suitability of the composition.

[0005] To achieve the above objectives, the present invention provides a whitening and spot-fading composition containing Polygonum cuspidatum-derived vesicles, comprising the following components by mass percentage: 0.001%-5% Polygonum cuspidatum-derived vesicles, 0.1%-15% whitening active ingredients, 0.01%-8% soothing and repairing ingredients, 1%-30% moisturizer, 0.01%-3% thickener, 0.01%-3% antioxidant, 0.05%-3% preservative system, 0.001%-1% pH adjuster, and the balance being water.

[0006] Furthermore, Polygonum cuspidatum-derived vesicles refer to nanoscale membranous vesicle-like raw materials derived from Polygonum cuspidatum plant materials. The Polygonum cuspidatum plant materials can be selected from the roots, rhizomes, stems, leaves, or combinations thereof, preferably the roots and / or rhizomes.

[0007] Furthermore, vesicle-like nanoparticles derived from Polygonum cuspidatum can be obtained through conventional separation and purification techniques in the field of plant vesicles, including tissue disruption, centrifugation to remove impurities, membrane filtration, ultrafiltration concentration, size exclusion chromatography, density gradient centrifugation, ultracentrifugation, tangential flow filtration, or any combination thereof.

[0008] Furthermore, the mass percentage of Polygonum cuspidatum-derived vesicles in the skin whitening and spot-fading composition is 0.01%-2%.

[0009] Preferably, the mass percentage of Polygonum cuspidatum-derived vesicles in the whitening and spot-fading composition is 0.05%-1%.

[0010] Furthermore, the vesicles derived from Polygonum cuspidatum are membrane-like vesicles with a particle size distribution mainly between 30 nm and 300 nm.

[0011] Preferably, the particle size distribution of Polygonum cuspidatum-derived vesicles is 50nm-500nm.

[0012] Furthermore, the whitening active ingredients are used to improve dull skin, assist in regulating melanin production or transport, and together with Polygonum cuspidatum-derived vesicles, they form a complex whitening skincare system.

[0013] Furthermore, the whitening active ingredients are selected from one or more of the following: niacinamide, tranexamic acid, α-arbutin, 3-O-ethyl ascorbic acid, ascorbate glucoside, ascorbate phosphate salt, glutathione, undecenoyl phenylalanine, licorice root extract, and tranexamic acid derivatives.

[0014] Preferably, the whitening active ingredients include 1%-8% niacinamide and / or 0.1%-3% tranexamic acid; Preferably, the whitening active ingredients also include 0.05%-3% of 3-O-ethyl ascorbic acid and / or 0.05%-2% of α-arbutin.

[0015] Furthermore, the soothing and repairing components enhance the skin suitability of the whitening and spot-fading composition and help improve skin discomfort caused by dryness, external irritation, or weakened barrier function.

[0016] Furthermore, the soothing and repairing components are selected from one or more of panthenol, allantoin, dipotassium glycyrrhizate, beta-glucan, ectoine, centella asiatica extract, purslane extract, bisabolol, ceramide, cholesterol, and phytosphingosine.

[0017] Preferably, the soothing and repairing components include one or more of panthenol (0.1%-5%), dipotassium glycyrrhizate (0.01%-1%), allantoin (0.01%-0.5%), and ectoine (0.01%-2%).

[0018] Furthermore, moisturizers are used to improve the moisturizing properties and skin feel of the whitening and spot-fading composition.

[0019] Furthermore, the moisturizer is selected from one or more of glycerin, butylene glycol, 1,3-propanediol, propylene glycol, 1,2-hexanediol, sodium hyaluronate, betaine, trehalose, sodium PCA, sorbitol, and erythritol.

[0020] Preferably, the moisturizer includes one or more of the following: glycerin 1%-10%, butylene glycol 1%-10%, 1,3-propanediol 1%-10%, betaine 0.1%-5%, and sodium hyaluronate 0.005%-0.5%.

[0021] Furthermore, thickeners are used to adjust the viscosity, rheology, and system stability of the whitening and spot-fading composition.

[0022] Furthermore, the thickener is selected from one or more of xanthan gum, hydroxyethyl cellulose, carbomer, ammonium acryloyl dimethyl taurate / VP copolymer, hydroxyethyl acrylate / sodium acryloyl dimethyl taurate copolymer, and sodium hyaluronate.

[0023] Preferably, the thickener includes one or more of xanthan gum (0.01%-0.5%), hydroxyethyl cellulose (0.05%-1%), carbomer (0.05%-1%), and ammonium acryloyldimethyl taurate / VP copolymer (0.05%-2%).

[0024] Furthermore, antioxidants are used to improve the antioxidant stability and formulation tolerance of the skin whitening and spot-fading compositions.

[0025] Furthermore, the antioxidant is selected from one or more of p-hydroxyacetophenone, tocopherol, carnosine, ferulic acid, sodium phytate, and sodium metabisulfite.

[0026] Preferably, the antioxidants include 0.05%-1% p-hydroxyacetophenone and / or 0.01%-0.2% sodium phytate.

[0027] Furthermore, the preservative system is used to ensure the microbial safety of the whitening and spot-fading composition during storage and use.

[0028] Furthermore, the preservative system is selected from one or more of phenoxyethanol, ethylhexylglycerin, 1,2-hexanediol, octyl glycol, methylparaben, ethylparaben, sodium benzoate, potassium sorbate, and capryloyl hydroxamic acid.

[0029] Preferably, the preservative system includes one or more of the following: phenoxyethanol 0.2%-1%, ethylhexylglycerin 0.05%-0.5%, 1,2-hexanediol 0.1%-2%, and octyl glycol 0.01%-0.5%.

[0030] Furthermore, a pH adjuster is used to adjust the whitening and spot-fading composition to a pH range suitable for topical skin application and conducive to system stability.

[0031] Furthermore, the pH adjuster is selected from one or more of citric acid, sodium citrate, lactic acid, sodium lactate, arginine, sodium hydroxide, triethanolamine, and aminomethylpropanol.

[0032] Furthermore, the pH of the whitening and spot-fading composition is 5.0-7.2.

[0033] Preferably, the pH of the whitening and spot-fading composition is 5.5-6.8.

[0034] Preferably, the whitening and spot-fading composition may further include emulsifiers, emollients, chelating agents, film-forming agents, fragrances, pigments, or other cosmetically acceptable excipients. Emollients may be selected from one or more of squalane, caprylic / capric triglycerides, jojoba seed oil, isononyl isononanoate, polydimethylsiloxane, and hydrogenated polyisobutylene; emulsifiers may be selected from one or more of glyceryl stearate, PEG-100 stearate, cetearyl alcohol, cetearyl glucoside, and polyglycerol emulsifiers.

[0035] Preferably, the whitening and spot-fading composition is an aqueous system, an oil-in-water emulsion system, a gel system, an emulsion system, a cream system, or a freeze-dried compound system. The whitening and spot-fading composition can be prepared into topical skin products such as serums, emulsions, creams, gels, facial masks, sprays, freeze-dried powders, freeze-dried essences, or ampoules.

[0036] Preferably, the whitening and spot-fading composition includes: 0.05%-1% Polygonum cuspidatum-derived vesicles, 1%-5% nicotinamide, 0.5%-3% tranexamic acid, 0.1%-3% panthenol, 0.02%-0.5% dipotassium glycyrrhizate, 2%-8% glycerin, 2%-8% butylene glycol, 0.01%-0.2% sodium hyaluronate, 0.05%-1% thickener, 0.05%-1% antioxidant, 0.2%-2% preservative system; an appropriate amount of pH adjuster, and the remainder being water.

[0037] Preferably, the whitening and spot-fading composition includes: 0.01%-2% Polygonum cuspidatum-derived vesicles, 0.1%-3% 3-O-ethyl ascorbic acid, 0.1%-2% α-arbutin, 0.01%-2% ectoine, 0.01%-0.5% allantoin, 3%-20% moisturizer, 0.05%-2% thickener, 0.05%-1% antioxidant, 0.2%-2% preservative system; an appropriate amount of pH adjuster, and the remainder being water.

[0038] On the other hand, the present invention also provides a method for preparing the above-mentioned skin whitening and spot-fading composition containing Polygonum cuspidatum-derived vesicles, comprising the following steps: S1. Add water to the mixing container, and under stirring conditions, add the humectant, chelating agent, and / or part of the thickener in sequence to fully disperse or dissolve them, thereby obtaining an aqueous matrix. If necessary, the system can be heated to 40℃-80℃ to promote the hydration of the thickener or the dissolution of water-soluble raw materials; for systems that can dissolve without heating, the aqueous matrix can also be prepared at room temperature.

[0039] S2. Add the whitening active ingredient to the aqueous matrix obtained in S1, and stir until it is evenly dispersed or completely dissolved. For heat-sensitive or easily oxidized whitening active ingredients, it is preferable to add them at a temperature not exceeding 45°C; for components that require heating to dissolve, dissolution can be carried out within a temperature range that does not affect their stability and formulation compatibility.

[0040] S3. Add the soothing and repairing components, antioxidants, preservatives, remaining thickeners, and other cosmetically acceptable excipients to the system obtained in S2, and stir until the system is homogeneous.

[0041] S4. Cool the system obtained in S3 to no higher than 45°C, preferably no higher than 40°C, and more preferably 15°C-35°C. Then add Polygonum cuspidatum-derived vesicles and mix them under low shear conditions to make them evenly dispersed in the whitening and spot-fading composition.

[0042] Low-shear conditions include paddle stirring, anchor stirring, magnetic stirring, low-speed homogenization, or a combination thereof; the stirring speed is preferably 50 rpm-800 rpm, more preferably 100 rpm-500 rpm; the stirring time is preferably 5 min-60 min, more preferably 10 min-30 min. By adding Polygonum cuspidatum-derived vesicles under lower temperature and low-shear conditions, the influence of high temperature or strong shear on the dispersion state and particle size stability of the vesicle-like raw materials during processing can be reduced.

[0043] S5. Add a pH adjuster to the system obtained in S4 to adjust the pH of the system to 5.0-7.2, preferably 5.5-6.8; then perform defoaming, settling, filtration, low-speed homogenization and / or filling to obtain a whitening and spot-fading composition containing Polygonum cuspidatum-derived vesicles. Filtration is preferably performed using a filtration method suitable for cosmetic production to remove undispersed particles or impurities; for products containing high viscosity systems or emulsion systems, terminal filtration may be omitted, and defoaming can be performed directly for filling.

[0044] Furthermore, when the whitening and spot-fading composition is an aqueous essence, facial mask, spray, or ampoule, S1-S5 can be completed in an aqueous system. It is preferable to avoid high-temperature sterilization, long-term high-speed homogenization, or strong ultrasonic treatment after adding Polygonum cuspidatum-derived vesicles.

[0045] Furthermore, when the whitening and spot-fading composition is an emulsion or cream, the aqueous phase and oil phase can be prepared separately first, and the aqueous phase and oil phase can be emulsified to form a basic emulsion system. After cooling, the heat-sensitive components, soothing and repairing components, and Polygonum cuspidatum-derived vesicles from the whitening active ingredients can be added.

[0046] Preferably, the Polygonum cuspidatum-derived vesicles are added after emulsification is complete and the temperature is cooled to no higher than 45°C.

[0047] Furthermore, when the whitening and spot-fading composition is a freeze-dried powder, freeze-dried essence, or freeze-dried compound system, the Polygonum cuspidatum-derived vesicles can be mixed with freeze-drying protectants, some moisturizers, and stabilizers and then freeze-dried. When used, it can be compounded with a solvent containing whitening active ingredients; or the liquid composition containing Polygonum cuspidatum-derived vesicles can be dispensed under low-temperature conditions and then freeze-dried.

[0048] Preferably, the freeze-drying protectant is selected from one or more of trehalose, mannitol, sucrose, glycine, and sodium hyaluronate.

[0049] Compared with the prior art, the present invention has the following advantages and technical effects: (1) The whitening and spot-fading composition provided by the present invention uses Polygonum cuspidatum-derived vesicles as one of the functional skin care raw materials, and is compounded with one or more whitening active ingredients, while being supplemented with soothing and repairing ingredients, moisturizers, antioxidants and stabilizing matrix to form a complex whitening skin care formula system. This composition not only focuses on the links related to melanin production or transport, but also takes into account factors such as skin barrier status, oxidative stress and topical mildness, which is beneficial to improving the shortcomings of single whitening active ingredients in comprehensive care.

[0050] (2) This invention combines Polygonum cuspidatum-derived vesicles with whitening active ingredients such as niacinamide, tranexamic acid, ascorbic acid derivatives, α-arbutin, and licorice root extract. It can construct various external skin care products such as serums, face masks, gels, lotions, creams, sprays, ampoules, or freeze-dried compound systems according to different dosage forms and usage needs, thereby improving the formulation applicability and product development flexibility of this type of plant-derived vesicle-like raw material in whitening skin care products.

[0051] (3) The preparation method of the present invention arranges the vesicles derived from Polygonum cuspidatum to be added after the system is cooled down, and preferably uses low shear conditions for mixing and dispersion. This can reduce the potential impact of high temperature, long-term high-speed homogenization or strong shear processing on the particle size distribution, dispersion state and formulation compatibility of nanoscale membrane vesicle-like raw materials, which is beneficial to improving the formulation stability and quality consistency of the whitening and spot-fading composition.

[0052] (4) The present invention sets moisturizer, soothing and repairing components, antioxidant, preservative system and pH adjustment system in the formulation system, which can improve the comfort of use, storage stability and microbial safety of the composition while meeting the application requirements of whitening and skin care, making it more suitable for development into daily skin care products. Attached Figure Description

[0053] To more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the drawings used in the embodiments will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For those skilled in the art, other drawings can be obtained based on these drawings without creative effort.

[0054] Figure 1 This is a particle size distribution characterization diagram of vesicle-like raw materials derived from Polygonum cuspidatum; Figure 2 TEM characterization of vesicle-like raw materials derived from Polygonum cuspidatum; Figure 3 A schematic diagram of the preparation process of a skin-whitening and spot-fading composition containing Polygonum cuspidatum-derived vesicles. Detailed Implementation

[0055] Various exemplary embodiments of the present invention will now be described in detail. This detailed description should not be considered as a limitation of the present invention, but rather as a more detailed description of certain aspects, features, and embodiments of the present invention.

[0056] It should be understood that the terminology used in this invention is merely for describing particular embodiments and is not intended to limit the invention. Furthermore, with respect to numerical ranges in this invention, it should be understood that each intermediate value between the upper and lower limits of the range is also specifically disclosed. Every smaller range between any stated value or intermediate value within a stated range, and any other stated value or intermediate value within said range, is also included in this invention. The upper and lower limits of these smaller ranges may be independently included or excluded from the range.

[0057] Unless otherwise stated, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art. While only preferred methods and materials have been described herein, any methods and materials similar or equivalent to those described herein may be used in the implementation or testing of this invention. All references to this specification are incorporated by way of citation to disclose and describe methods and / or materials associated with those references. In the event of any conflict with any incorporated reference, the content of this specification shall prevail.

[0058] Various modifications and variations can be made to the specific embodiments described in this specification without departing from the scope or spirit of the invention, as will be apparent to those skilled in the art. Other embodiments derived from this specification will also be apparent to those skilled in the art. This specification and embodiments are merely exemplary.

[0059] The terms “include,” “including,” “have,” “contain,” etc., used in this article are all open-ended terms, meaning that they include but are not limited to.

[0060] Unless otherwise specified, the term "parts" in this invention refers to parts by weight.

[0061] Polygonum cuspidatum-derived vesicles refer to nanoscale membrane-like vesicle-like raw materials derived from Polygonum cuspidatum plant materials. Polygonum cuspidatum plant materials can be selected from the roots, rhizomes, stems, leaves, or combinations thereof.

[0062] Vesicle-like nanoparticles derived from Polygonum cuspidatum are not limited to a specific preparation process and can be obtained through conventional separation and purification techniques in the field of plant vesicles, including tissue disruption, centrifugation for impurity removal, membrane filtration, ultrafiltration concentration, size exclusion chromatography, density gradient centrifugation, ultracentrifugation, tangential flow filtration, or any combination thereof. As long as the obtained vesicle-like particles are derived from Polygonum cuspidatum and possess the membrane vesicle structure, nanoparticle size range, and oral mucosal repair-related bioactivity defined in this invention, they fall within the scope of this invention.

[0063] Example 1 The preparation method of Polygonum cuspidatum-derived vesicles includes the following steps: (1) Raw material pretreatment: Take fresh Polygonum cuspidatum roots, remove surface soil and impurities, wash repeatedly with deionized water, drain the surface water and set aside. Freshly collected Polygonum cuspidatum roots and rhizomes can be used as materials. After harvesting, they should be stored at a low temperature of 4℃ and the extraction and preparation should be completed within 24 hours to reduce the degradation of active ingredients.

[0064] (2) Tissue disruption: The cleaned roots and rhizomes of Polygonum cuspidatum were added to pre-cooled phosphate-buffered saline (PBS, pH 7.4, 4℃) at a material-to-liquid ratio of 1:3 g / mL, and disrupted at high speed using a tissue homogenizer or a high-speed blender. The disruption conditions were: 12000 rpm, disruption time 5 min, and the disruption process was carried out under ice bath conditions to control the system temperature not to exceed 4℃. The resulting material was a Polygonum cuspidatum homogenate.

[0065] (3) Low-speed centrifugation to remove fibers and cell debris: Transfer the above homogenate to a centrifuge tube and centrifuge at 4°C at low speed. Centrifugation conditions: 2000×g, centrifugation time 15min. After centrifugation, collect the supernatant and discard the precipitate. Centrifuge the supernatant again at 4°C and 8000×g for 25min and collect the supernatant.

[0066] (4) Removal of large particulate impurities by membrane filtration: The supernatant after low-speed centrifugation was successively passed through filter membranes with pore sizes of 1.0 μm and 0.45 μm for microfiltration to remove large particulate impurities and suspended solids, and the filtrate was collected. The filtration operation was carried out at 4℃, and the filter membrane material was polyethersulfone (PES).

[0067] (5) Ultrafiltration Concentration and Purification: The above filtrate was transferred to an ultrafiltration concentration system and concentrated using an ultrafiltration membrane with a molecular weight cutoff of 100 kDa. The operating parameters were: inlet pressure 0.2 MPa, permeation pressure 0.1 MPa, and operating temperature 15 °C. During ultrafiltration, molecules smaller than 100 kDa were discharged with the permeate, while larger vesicle-like nanoparticles were retained and concentrated. When the concentration was reduced to 1 / 15 of the original volume, the concentrate was collected. An equal volume of PBS was added to the concentrate for washing (percolation), and this process was repeated three times to improve vesicle purity.

[0068] (6) Further purification: The ultrafiltration concentrate was purified by sucrose density gradient centrifugation. A discontinuous sucrose density gradient (8%, 30%, 45%, 60%, w / v) was prepared. The concentrate was spread on top of the gradient and ultracentrifuged at 4℃ and 100,000×g for 90 min. After centrifugation, the vesicle bands located between the 8%-30% sucrose density layers were collected, diluted with PBS, and then ultrafiltered again for desalting and concentration to obtain purified Polygonum cuspidatum-derived vesicles.

[0069] The prepared Polygonum cuspidatum-derived vesicle concentrate was aliquoted into sterile cryovials and stored at -80°C for later use. Before use, it was diluted with PBS to the required concentration.

[0070] The particle size distribution of the obtained Polygonum cuspidatum-derived vesicles was analyzed by NTA, and the results are as follows: Figure 1 As shown, TEM observation revealed a membrane-bound vesicle structure, such as... Figure 2 As shown. The obtained particles, as determined by NTA, have a particle size distribution of 161.6 ± 84.7 nm. TEM observation revealed a membrane vesicle structure with a zeta potential of -49.4 mV.

[0071] Example 2 A whitening essence containing Polygonum cuspidatum-derived vesicles, comprising the following components by weight percentage: Polygonum cuspidatum-derived vesicles 1%, niacinamide 4.0%, tranexamic acid 1.0%, 3-O-ethyl ascorbic acid 0.5%, panthenol 1.0%, dipotassium glycyrrhizate 0.1%, allantoin 0.1%, glycerin 5.0%, butylene glycol 5.0%, 1,2-hexanediol 0.5%, sodium hyaluronate 0.05%, xanthan gum 0.1%, p-hydroxyacetophenone 0.4%, ethylhexylglycerin 0.1%; a suitable amount of pH adjuster; the balance being water.

[0072] The preparation process of whitening essence is as follows: Figure 3 As shown, it includes the following steps: S1. Add water to the mixing container, and add glycerin, butylene glycol, sodium hyaluronate and xanthan gum under stirring conditions. Stir until sodium hyaluronate and xanthan gum are fully dispersed or hydrated to obtain an aqueous matrix.

[0073] S2. Add nicotinamide, tranexamic acid and 3-O-ethyl ascorbic acid to the aqueous matrix obtained in S1, and stir until dissolved or uniformly dispersed.

[0074] S3. Add panthenol, dipotassium glycyrrhizate, allantoin, p-hydroxyacetophenone, 1,2-hexanediol and ethylhexylglycerin to the system obtained in S2, and continue stirring until the system is homogeneous.

[0075] S4. Control the temperature of the system obtained in S3 to no higher than 40℃, add Polygonum cuspidatum-derived vesicles, and mix them using a low-shear method to ensure that they are evenly dispersed in the system.

[0076] S5. Add a pH adjuster to adjust the pH of the system to 5.8-6.5. After degassing, fill the bottle to obtain a whitening essence containing Polygonum cuspidatum-derived vesicles.

[0077] The whitening essence obtained in this embodiment is suitable for daily external skin care and can be used in skin care scenarios such as whitening, brightening, improving dullness, and assisting in soothing and repairing.

[0078] Example 3 A skin-brightening facial mask liquid containing Polygonum cuspidatum-derived vesicles, comprising the following components by weight percentage: Polygonum cuspidatum-derived vesicles 0.5%, niacinamide 3.0%, α-arbutin 0.5%, ascorbate glucoside 0.5%, β-glucan 0.2%, panthenol 0.8%, purslane extract 0.5%, glycerin 4.0%, 1,3-propanediol 5.0%, betaine 2.0%, hydroxyethyl cellulose 0.2%, p-hydroxyacetophenone 0.4%, 1,2-hexanediol 0.5%; a suitable amount of pH adjuster; the balance being water.

[0079] The preparation method of the brightening facial mask liquid includes the following steps: S1. Add water to the mixing container, and add glycerol, 1,3-propanediol, betaine and hydroxyethyl cellulose under stirring conditions. Stir until the hydroxyethyl cellulose is fully dispersed or hydrated to obtain an aqueous matrix.

[0080] S2. Add nicotinamide, α-arbutin and ascorbate glucoside to the aqueous matrix obtained in S1, and stir until the system is homogeneous.

[0081] S3. Add β-glucan, panthenol, purslane extract, p-hydroxyacetophenone and 1,2-hexanediol to the system obtained in S2, and continue stirring until homogeneous.

[0082] S4. Control the system temperature to no higher than 40℃, add Polygonum cuspidatum-derived vesicles, and mix evenly using paddle stirring, anchor stirring, or other low-shear stirring methods.

[0083] S5. Adjust the pH of the system to 5.5-6.8, and after degassing, filtration or direct filling, obtain a brightening facial mask liquid containing vesicles derived from Polygonum cuspidatum.

[0084] The brightening mask liquid obtained in this embodiment can be used in conjunction with non-woven fabric, silk mask cloth, bio-cellulose mask cloth or other cosmetically acceptable mask carriers, and can also be used as a topical brightening care liquid.

[0085] Example 4 A whitening lotion containing Polygonum cuspidatum-derived vesicles, comprising the following components by weight percentage: Polygonum cuspidatum-derived vesicles 0.2%, niacinamide 2.0%, tranexamic acid 0.5%, Glycyrrhiza glabra root extract 0.2%, glycerin 4.0%, butylene glycol 3.0%, squalane 2.0%, caprylic / capric triglyceride 3.0%, glyceryl stearate 1.0%, cetearyl alcohol 1.0%, panthenol 0.5%, ceramide 0.05%, cholesterol 0.05%, carbomer 0.15%, triethanolamine as needed; preservative system as needed; balance water.

[0086] The preparation method of whitening lotion includes the following steps: S1. Add water, glycerol, butylene glycol and carbomer to an aqueous phase pot, stir and disperse, and heat if necessary to fully hydrate it to obtain an aqueous phase.

[0087] S2. Add squalane, caprylic / capric triglyceride, glyceryl stearate and cetearyl alcohol to the oil phase pot, heat and stir until completely dissolved to obtain the oil phase.

[0088] S3. Add the oil phase to the aqueous phase and emulsify to form a basic emulsion system.

[0089] S4. After cooling the basic emulsified system obtained in S3, add nicotinamide, tranexamic acid, licorice root extract, panthenol, ceramide, cholesterol, and preservative system, and stir until homogeneous.

[0090] S5. When the system temperature drops to no higher than 40℃, add Polygonum cuspidatum-derived vesicles and mix them evenly using a low-speed stirring method.

[0091] S6. Add triethanolamine or other pH adjusters to adjust the pH of the system to 5.5-6.8, degas and fill to obtain a whitening emulsion containing Polygonum cuspidatum-derived vesicles.

[0092] In this embodiment, the Polygonum cuspidatum-derived vesicles are added after emulsification and cooling, which can reduce the impact of high temperature and high shear conditions during the emulsification stage on the dispersion state of the vesicle-like raw materials.

[0093] Example 5 A skin-whitening gel containing Polygonum cuspidatum-derived vesicles, comprising the following components by weight percentage: 0.5% Polygonum cuspidatum-derived vesicles, 2.0% niacinamide, 1.0% tranexamic acid, 0.2% ectoine, 1.0% panthenol, 0.05% dipotassium glycyrrhizate, 4.0% glycerin, 4.0% 1,3-propanediol, 1.0% betaine, 0.2% carbomer, 0.05% xanthan gum, 0.4% p-hydroxyacetophenone, and 0.5% 1,2-hexanediol; a suitable amount of pH adjuster; and the balance being water.

[0094] The preparation method of whitening gel includes the following steps: S1. Mix water, glycerin, 1,3-propanediol and betaine, and add carbomer and xanthan gum under stirring to fully disperse and hydrate them.

[0095] S2. Add nicotinamide, tranexamic acid, ectoine, panthenol, dipotassium glycyrrhizate, p-hydroxyacetophenone and 1,2-hexanediol, and stir until the system is homogeneous.

[0096] S3. Control the system temperature to no higher than 40℃, add Polygonum cuspidatum-derived vesicles, and stir at low speed until uniformly dispersed.

[0097] S4. Add a pH adjuster to neutralize and thicken, adjust the pH of the system to 5.5-6.8, and fill after degassing to obtain a whitening gel containing Polygonum cuspidatum-derived vesicles.

[0098] Example 6 A freeze-dried compound system containing Polygonum cuspidatum-derived vesicles includes freeze-dried components and a redissolving solvent.

[0099] The freeze-dried components, by mass percentage, include: Polygonum cuspidatum vesicles 2.0%; trehalose 2%; mannitol 1.0%; sodium hyaluronate 0.05%; and the balance being water.

[0100] The reconstituted solvent includes 4% nicotinamide, 1% tranexamic acid, 2% panthenol, 8% humectant, an appropriate amount of preservative system, and water.

[0101] The preparation method of the freeze-dried compound system includes the following steps: S1. Mix the vesicles derived from Polygonum cuspidatum with trehalose, mannitol, sodium hyaluronate, and water, and stir at low temperature and low shear until homogeneous.

[0102] S2. The mixture obtained in S1 is dispensed and then freeze-dried to obtain a freeze-dried component containing vesicles derived from Polygonum cuspidatum.

[0103] S3. Before use, mix the freeze-dried components with a reconstituted solvent containing whitening active ingredients. After reconstitution, a whitening skin care composition containing Polygonum cuspidatum-derived vesicles is obtained.

[0104] In this embodiment, trehalose, mannitol, and sodium hyaluronate can be used as freeze-drying protectants or stabilizers to help maintain the dispersion state of the system during freeze-drying and reconstitution. The specific freeze-drying procedure can be routinely optimized according to equipment conditions, fill volume, and product specifications.

[0105] Test Example 1 The experiment was conducted according to the group standard T / GDCA 006-2021, "In vitro tyrosinase activity inhibition test method". Mushroom tyrosinase (Sigma-Aldrich, catalog number T3824) was used as the enzyme source, and L-DOPA as the substrate. The experiment was performed in 96-well plates, with the following groups included: blank control group, model control group, positive control group (kojic acid, 100 μg / mL), basic formula group (whitening and spot-fading composition matrix without Polygonum cuspidatum-derived vesicles), Polygonum cuspidatum-derived vesicle group (1% Polygonum cuspidatum-derived vesicle aqueous solution), and whitening and spot-fading composition group containing Polygonum cuspidatum-derived vesicles (whitening essence obtained in Example 2, tested at three concentrations: 1%, 5%, and 10%). Each experimental group had three replicates.

[0106] The reaction system was as follows: phosphate-buffered saline (PBS, pH 6.8), the sample to be tested, and tyrosinase solution (200 U / mL) were added to each well. After incubation at 37°C for 10 min, L-DOPA substrate solution (2 mg / mL) was added, and the reaction was continued at 37°C for another 20 min. After the reaction, the absorbance (OD) of each well was measured at a wavelength of 475 nm. 475 For samples that have color or absorbent background, set up a corresponding sample background subtraction group (using PBS instead of tyrosinase solution).

[0107] The inhibition rate of tyrosinase activity is calculated using the following formula: .

[0108] The experimental results are shown in Table 1 below.

[0109] Table 1 Results of Tyrosinase Inhibition Rate Measurement

[0110] Experimental results show that Polygonum cuspidatum-derived vesicles, when used alone, have a certain inhibitory effect on tyrosinase (inhibition rate 23.5%), which is related to its active ingredients such as resveratrol and polygalactosidase glycoside. The basic formula group (without Polygonum cuspidatum-derived vesicles) showed a tyrosinase inhibition rate of only 12.3%, mainly due to the whitening active ingredients such as nicotinamide and tranexamic acid in the formula. The whitening and spot-fading composition containing Polygonum cuspidatum-derived vesicles showed significantly higher tyrosinase inhibitory activity than the sum of the individual components at all concentrations, suggesting a possible synergistic effect between Polygonum cuspidatum-derived vesicles and whitening active ingredients. When the composition was tested at a concentration of 10%, the inhibition rate reached 78.5%, close to the level of the positive control kojic acid (85.6%). The above results indicate that the whitening and spot-fading composition containing Polygonum cuspidatum-derived vesicles provided by this invention can reduce melanin production from the source by inhibiting tyrosinase activity, thereby exerting a whitening and spot-fading effect.

[0111] Test Example 2 Mouse melanoma cells B16F10 (ATCC CRL-6475) were used as an in vitro cell model. Cells were cultured in DMEM high-glucose medium containing 10% fetal bovine serum (FBS) and 1% penicillin-streptomycin at 37°C in a 5% CO2 incubator.

[0112] The experiment included a blank control group (containing only culture medium), a model control group (containing 100 nM α-MSH stimulation), a positive control group (containing 100 nM α-MSH + 100 μg / mL arbutin), a basic formula group, a Polygonum cuspidatum-derived vesicle group (50 μg / mL), and a whitening and spot-fading composition group containing Polygonum cuspidatum-derived vesicles (the whitening essence obtained in Example 2, tested at three concentrations of 0.5%, 1%, and 2%). Each group had three replicates.

[0113] Take B16F10 cells in the logarithmic growth phase and use 1×10 4 Cells / well were seeded at a density of [number] cells / well in 96-well plates and cultured for 24 hours. The culture medium was then replaced with fresh medium containing the corresponding test substance (100 nM α-MSH), and cultured for another 72 hours. After culture: (1) Cell viability assay: Cell viability was detected by the CCK-8 assay to exclude false positive results caused by significant cytotoxicity. 10 μL of CCK-8 reagent was added to each well and incubated at 37°C for 2 h. The absorbance at 450 nm was then measured.

[0114] (2) Detection of intracellular melanin content: Discard the culture medium, wash twice with PBS, add 100 μL of 1M NaOH solution containing 10% DMSO, and heat in an 80℃ water bath for 30 min until the cells are completely lysed. Measure the absorbance of each well at a wavelength of 405 nm. At the same time, the protein content of each well is determined by the BCA method, and the melanin content is expressed as OD. 405 / mg protein is expressed as a percentage.

[0115] Melanin inhibition rate is calculated using the following formula: .

[0116] The measurement results are shown in Table 2 below.

[0117] Table 2 Results of melanin inhibition rate measurement

[0118] CCK-8 assay results showed that the cell viability of each test substance group was above 94%, with no significant difference compared with the blank control group (P>0.05), indicating that each test substance had no obvious cytotoxic effect within the test concentration range.

[0119] In the model control group, melanin content increased significantly under α-MSH induction (from 0.82 to 2.35 OD). 405The presence of [ / mg protein] indicates the successful construction of the melanin-producing cell model. The melanin inhibition rate of the basic formula group was 21.3%, mainly due to the inhibitory effects of ingredients such as nicotinamide and tranexamic acid. The inhibition rate of Polygonum cuspidatum-derived vesicles alone was 28.5%, consistent with its inhibitory trend on tyrosinase in Test Example 1. The whitening and spot-fading composition containing Polygonum cuspidatum-derived vesicles achieved an inhibition rate of 47.7% at a 1% concentration and 55.3% at a 2% concentration, exceeding the level of the positive control arbutin (52.3%). These results indicate that the combination of Polygonum cuspidatum-derived vesicles and whitening active ingredients can exert a whitening effect by inhibiting melanin production in melanocytes, and its effect is superior to that of a single component.

[0120] Test Example 3 The antioxidant capacity of the skin-lightening and spot-fading compositions was evaluated using the DPPH (1,1-diphenyl-2-trinitrophenylhydrazine) free radical scavenging method. Different concentrations of the test sample (the skin-lightening essence obtained in Example 2, with concentration gradients of 0.1%, 0.5%, 1%, 2%, and 5%) were added to 96-well plates and mixed with 200 μmol / L DPPH ethanol solution (1:1, v / v). After reacting at room temperature in the dark for 30 min, the absorbance was measured at 517 nm. Vitamin C (VC) was used as a positive control, and deionized water was used as a blank control. Each experimental group had three replicates.

[0121] The results of the DPPH free radical scavenging rate determination are shown in Table 3 below.

[0122] Table 3 Results of DPPH free radical scavenging rate determination

[0123] Experimental results showed that the skin-whitening and spot-fading composition containing Polygonum cuspidatum-derived vesicles exhibited significant DPPH free radical scavenging ability, showing a good dose-dependent relationship. At a 5% concentration, the DPPH scavenging rate reached 81.3%, close to the level of the vitamin C positive control (92.5%). The resveratrol and polygalactosidase active ingredients in the Polygonum cuspidatum-derived vesicles possess strong antioxidant activity, working synergistically with antioxidants such as p-hydroxyacetophenone and tocopherol in the formulation to give the composition excellent free radical scavenging ability. Antioxidant activity is one of the important auxiliary mechanisms for skin whitening and spot fading; by scavenging free radicals, it can reduce the promoting effect of oxidative stress on melanin production, while also helping to protect skin cells from oxidative damage.

[0124] Test Example 4 The test was conducted according to the group standard T / SHRH 036-2021 "Melanin Inhibition Test in Cosmetics - Zebrafish Embryo Test Method". Normally developing zebrafish (Danio rerio) embryos (6-8 hours post-fertilization) were randomly divided into a blank control group (standard culture medium), a model control group, a positive control group (arbutin 100 μg / mL), a basic formula group, a Polygonum cuspidatum-derived vesicle group (50 μg / mL), and a whitening and spot-fading composition group containing Polygonum cuspidatum-derived vesicles (the whitening essence obtained in Example 2, tested at three concentrations: 0.5%, 1%, and 2%). Each group contained 20 embryos, and the experiment was repeated three times.

[0125] Zebrafish embryos were placed in 6-well plates, and 5 mL of culture medium containing the corresponding test substance was added to each well. The plates were incubated at 28°C for 72 hours post-fertilization. After incubation: (1) Observe and photograph the whole image of zebrafish under a stereomicroscope, and use image analysis software (ImageJ) to quantitatively analyze the melanin area and melanin gray value on the zebrafish body surface.

[0126] (2) The zebrafish embryos were homogenized and melanin and tyrosinase were extracted. The protein concentration was determined by BCA method, and the melanin content and tyrosinase activity were measured at 405 nm and 475 nm, respectively.

[0127] (3) The expression levels of melanin production-related genes mitfa, tyr, tyrp1, and dct were detected by real-time quantitative PCR (qPCR). At the same time, the mortality rate, deformity rate, hatching rate, and heart rate of zebrafish in each experimental group were observed to evaluate the safety of the samples.

[0128] The results of the zebrafish melanin inhibition rate determination are shown in Table 4 below.

[0129] Table 4 Results of the inhibition rate of melanin in zebrafish

[0130] The high transparency of zebrafish embryos allows for direct observation and quantitative analysis of melanin changes. In the model control group, without intervention from the test substance, the melanin area was significantly increased (3.62 mm). 2 The grayscale value decreased (62.1), indicating that the zebrafish melanin model was successfully constructed. The survival rate of zebrafish in each test group was above 95%, with no significant difference compared to the blank control group. P >0.05), indicating that each test substance has no significant toxicity to zebrafish embryos within the tested concentration range.

[0131] A skin-lightening and blemish-reducing composition containing Polygonum cuspidatum-derived vesicles showed an inhibition rate of 42.5% on melanin production in zebrafish at a 2% concentration, approaching the level of the positive control arbutin (44.5%). Simultaneously, qPCR results showed that the skin-lightening and blemish-reducing composition containing Polygonum cuspidatum-derived vesicles significantly downregulated the expression levels of melanin production-related genes such as mitfa, tyr, tyrp1, and dct. P The concentration of melanin in the sample was <0.05%, further confirming its mechanism of action in inhibiting melanin production at the molecular level. These results demonstrate that the composition provided by this invention exhibits significant skin-whitening and spot-fading effects at the whole animal level.

[0132] Test Example 5 Subject to relevant ethical and regulatory requirements, the whitening and spot-fading effects of the whitening and spot-fading composition were tested using human efficacy evaluation methods.

[0133] We are recruiting 30 healthy female volunteers aged 25-50 with facial pigmentation or dull skin tone, but no history of serious skin diseases or cosmetic allergies. A randomized, double-blind, half-face controlled trial design will be used.

[0134] Subjects used the whitening essence containing Polygonum cuspidatum-derived vesicles prepared in Example 2 (test side) and the basic formula essence (control side) twice daily, morning and evening, for 8 consecutive weeks. At week 0, 4 weeks, and 8 weeks after use, the L* value (skin brightness), ITA° value (individual type angle), and melanin index (MI) of the skin on both cheeks of the subjects were measured using a skin colorimeter. Simultaneously, facial images of the subjects were captured using a standardized facial image acquisition system, and changes in the area and depth of pigmentation were assessed using image analysis software. Adverse reactions such as erythema, itching, stinging, dryness, and desquamation were recorded concurrently. The experimental results are shown in Table 5 below.

[0135] Table 5 Results of Human Efficacy Evaluation

[0136] Note: * indicates comparison with the period before use (week 0) on the same side. P <0.05, ** indicates a difference compared to the level before use on the same side (week 0). P <0.01; # indicates that the test side is compared with the control side at the same time point. P <0.05.

[0137] After 8 weeks of continuous use, the L* value of the test side skin significantly increased from 58.2 to 64.3. P <0.01), the ITA° value significantly increased from 38.5 to 47.6 ( P <0.01 indicates a significant improvement in skin brightness and a noticeable brightening of skin tone. The melanin index (MI) decreased significantly from 185.6 to 148.3 (P <0.01), a decrease of 20.1%, indicating a significant reduction in skin melanin content. No significant changes were observed in any indicators before and after use in the control group. P The concentration was >0.05, excluding interference from environmental factors such as seasonal changes. Image analysis results showed that the average area of ​​the pigmented spots on the test side decreased by approximately 18.5%, and the color depth of the spots significantly lightened. No adverse reactions such as erythema, itching, or stinging were observed throughout the experiment, indicating that the whitening and pigmentation-fading composition containing Polygonum cuspidatum-derived vesicles provided by this invention has good skin tolerability and safety.

[0138] The embodiments described above are merely preferred embodiments of the present invention and are not intended to limit the scope of the present invention. Various modifications and improvements made by those skilled in the art to the technical solutions of the present invention without departing from the spirit of the present invention should fall within the protection scope defined by the claims of the present invention.

Claims

1. A skin-whitening and spot-fading composition containing vesicles derived from Polygonum cuspidatum, characterized in that, By weight percentage, it includes the following components: Polygonum cuspidatum-derived vesicles 0.001%-5%, whitening active ingredients 0.1%-15%, soothing and repairing ingredients 0.01%-8%, moisturizers 1%-30%, thickeners 0.01%-3%, antioxidants 0.01%-3%, preservative system 0.05%-3%, pH adjuster 0.001%-1%, and the balance being water.

2. The skin-whitening and spot-fading composition containing Polygonum cuspidatum-derived vesicles according to claim 1, characterized in that, The preparation method of Polygonum cuspidatum-derived vesicles includes: washing Polygonum cuspidatum roots and / or stems and leaves, adding aqueous buffer solution for tissue disruption, centrifuging the resulting homogenate at low speed to remove fibers and cell debris, then filtering through a membrane to remove large particulate impurities, and finally purifying by ultrafiltration to collect the component containing vesicle-like nanoparticles.

3. The whitening and spot-fading composition containing Polygonum cuspidatum-derived vesicles according to claim 1, characterized in that: The whitening active ingredients are selected from one or more of the following: nicotinamide, tranexamic acid, α-arbutin, 3-O-ethyl ascorbic acid, ascorbate glucoside, ascorbate phosphate salt, glutathione, undecenoyl phenylalanine, licorice root extract, and tranexamic acid derivatives.

4. The whitening and spot-fading composition containing Polygonum cuspidatum-derived vesicles according to claim 1, characterized in that: The soothing and repairing ingredients are selected from one or more of panthenol, allantoin, dipotassium glycyrrhizate, beta-glucan, ectoine, centella asiatica extract, purslane extract, bisabolol, ceramide, cholesterol, and phytosphingosine.

5. The whitening and spot-fading composition containing Polygonum cuspidatum-derived vesicles according to claim 1, characterized in that: The moisturizer is selected from one or more of glycerin, butylene glycol, 1,3-propanediol, propylene glycol, 1,2-hexanediol, sodium hyaluronate, betaine, trehalose, sodium PCA, sorbitol and erythritol.

6. The skin-whitening and spot-fading composition containing Polygonum cuspidatum-derived vesicles according to claim 1, characterized in that: The thickener is selected from one or more of xanthan gum, hydroxyethyl cellulose, carbomer, ammonium acryloyl dimethyl taurate / VP copolymer, hydroxyethyl acrylate / sodium acryloyl dimethyl taurate copolymer, and sodium hyaluronate; The antioxidant is selected from one or more of p-hydroxyacetophenone, tocopherol, carnosine, ferulic acid, sodium phytate, and sodium metabisulfite.

7. The skin-whitening and spot-fading composition containing Polygonum cuspidatum-derived vesicles according to claim 1, characterized in that: The composition has a pH of 5.0-8.

0.

8. A method for preparing a skin-whitening and spot-fading composition containing Polygonum cuspidatum-derived vesicles as described in any one of claims 1-7, characterized in that, The steps include the following: S1. Mix water, humectant, chelating agent and / or part of thickener to disperse or dissolve them to obtain an aqueous matrix; S2. Add the whitening active ingredients to the aqueous matrix of S1 and stir until homogeneous; S3. Add soothing and repairing ingredients, antioxidants, preservative system and remaining thickener, and mix thoroughly. S4. Control the system temperature to be no higher than 45℃, add Polygonum cuspidatum-derived vesicles, and mix using a low-shear method; S5. Adjust the pH to 5.0-7.5, defoam, filter or homogenize, and then fill to obtain the whitening and spot-fading composition.

9. The use of a whitening and spot-fading composition containing Polygonum cuspidatum-derived vesicles as described in any one of claims 1-7 in the preparation of skin care products having the effects of whitening, brightening, improving dull skin, fading pigmentation, soothing skin and / or assisting in repairing the skin barrier.

10. The application according to claim 9, characterized in that: Skincare products are one of the following: serum, lotion, cream, gel, facial mask liquid, spray, freeze-dried powder, freeze-dried essence, or ampoule.

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