A multiple anti-aging natural composition and skin care product thereof

Abstract

The present application belongs to the technical field of cosmetics, and particularly relates to a multiple anti-aging natural composition and a skin care product thereof. The present application prepares a hydrolyzed rice extract, a French wax chrysanthemum extract and a long-stem grape caulerpa lentillifera extract through a simple enzymatic process, and the three natural extracts are compounded in a specific ratio to form a composition which exhibits significant multiple anti-aging performance, can effectively promote the synthesis of collagen, periosteal protein, fibronectin and glycosaminoglycan, strengthen the structure and cross-linking of collagen fibers, restore and improve the structural integrity and functionality of ECM and DEJ, and can also effectively improve the moisturizing capacity and smoothness, thickness of the skin, realize the anti-aging effect through multiple pathways, and exhibit an obvious visible wrinkle-removing effect in the clinic.

Description

TECHNICAL FIELD

[0001] The present application belongs to the technical field of cosmetics, especially relates to the field under the IPC classification number A61K8 / 96, and more particularly relates to a multiple anti-aging natural composition and a skin care product thereof. BACKGROUND

[0002] With the growth of age, the skin of human gradually ages, and environmental factors such as air pollution, temperature, sun exposure can accelerate the aging of the skin. With the process of aging, the skin will produce intuitive and obvious changes in structure and function, such as wrinkles, impaired barrier function, slowed wound healing, etc. These aging characteristics reflect the changes in the composition, structure and function of the extracellular matrix (ECM).

[0003] The extracellular matrix is a kind of product secreted by cells in tissues and remaining in the extracellular, which is mainly composed of structural proteins, linking / adhesion proteins and amorphous gel-like structures. Among them, the structural proteins of dermal extracellular matrix are mainly type I and type III collagen. The collagen fiber network formed by type I collagen provides strong support to the skin, and the loss of type I collagen will appear facial wrinkles and depression. Type III collagen has good repair and healing promotion effect and promotes elasticity, which maintains the skin softness, delicate elasticity, etc.

[0004] There is also a key linking / adhesion protein in the extracellular matrix, periostin, which is a regenerative protein in the skin, participates in the regeneration of collagen fiber structure and consolidates the dermal tissue. With the aging of the skin, the synthesis of periostin decreases, leading to disordered or broken collagen fiber structure, resulting in loss of biomechanical properties, skin relaxation and wrinkles.

[0005] In addition, there are more than thirty kinds of proteoglycans in the dermal ECM, mainly glycosaminoglycans (GAGs), the amino polysaccharide side chain of which can effectively attract water, thereby forming a homogeneous gel-like filler to occupy the space of the network structure. With the aging of the skin, the synthesis of GAGs also decreases, and the water of the skin is more likely to be lost, the skin barrier becomes thinner, and the skin is more likely to be dry and wrinkled.

[0006] On the other hand, the dermal-epidermal junction (DEJ) is a key player in maintaining the cohesion of the skin related to the epidermis and dermis. Collagen IV constitutes the main component of the epidermal basement membrane, providing a framework for other molecules to maintain the functional interface between the epidermis and the dermis, and is also very important for maintaining the mechanical stability of the skin. The loss of collagen IV will also lead to the formation of wrinkles. Laminin-5 is the main adhesive component of DEJ. Studies have found that the synthesis of laminin-5 in aged skin decreases, and the contact between the epidermis and the dermis also decreases, leading to loss of skin elasticity and relaxation.

[0007] Therefore, there is an urgent need for a multi-faceted anti-aging solution to address skin aging from multiple dimensions such as ECM function enhancement, DEJ function interface repair, and dermal layer structure consolidation. The prior art CN 113230198A discloses an anti-wrinkle and anti-aging lyophilized essence containing sea grape extract, sturgeon seed extract, stem cell extract, green bean extract, hydrolyzed elastin, hydrolyzed rice protein and other active ingredients, but the essence only has antioxidant effect and can only prevent oxidation of free radicals on the skin to a certain extent, and cannot achieve repair and improvement of ECM and DEJ. SUMMARY

[0008] In view of the defects of the prior art, the purpose of the present application is to provide a multi-anti-aging natural composition with pure natural components, which can effectively enhance ECM function, promote collagen synthesis and fiber cross-linking, stimulate the production of periostin and fibronectin, and improve wrinkles.

[0009] The purpose of the present application is also to provide a skin care product containing the above-mentioned multi-anti-aging natural composition.

[0010] To achieve the above-mentioned purposes of the application, the following technical solutions are adopted:

[0011] A multi-anti-aging natural composition, comprising hydrolyzed rice extract, French Helichrysum Stoechas extract and Caulerpa Lentillidera extract.

[0012] Preferably, the mass ratio of the hydrolyzed rice extract, French Helichrysum Stoechas extract and Caulerpa Lentillidera extract is 0.5-2:0.5-4:0.5-2.

[0013] The present application selects a combination of hydrolyzed rice extract, French wax chrysanthemum extract and long-stem grape caulerpa extract to provide multi-channel anti-aging effects. The hydrolyzed rice extract has a significant promoting effect on the synthesis of collagen in dermal cells; the French wax chrysanthemum extract is rich in effective components such as oligosaccharides and can restore the synthesis function of periosteum protein, thereby repairing the aging and damaged collagen fiber structure, strengthening the function of the matrix network and the mechanical properties of the dermis, reducing the relaxation of the basement, and improving the tightness and elasticity of the skin; the long-stem grape caulerpa extract has a significant repairing effect on the functional interface of DEJ and can not only promote the production of collagen IV but also stimulate the expression of laminin-5, thicken the stratum corneum, and maintain and restore the structure and function of the epidermal basement membrane. The present inventors have also found that the multi-channel anti-aging natural composition of the present application can produce a synergistic effect on the synthesis promotion of collagen, laminin and glycosaminoglycan in a specific ratio. The promoting effect of the ternary combination is higher than the superposition of the promoting effects of single components or binary combinations. This may be because the same target triggers the overexpression of functional proteins when stimulated by the combined components, thereby further increasing the synthesis amount of these functional substances and achieving better anti-aging effects.

[0014] For example, the mass ratio of the hydrolyzed rice extract, the French wax chrysanthemum extract and the long-stem grape caulerpa extract is 1:0.5:1, 1.5:0.5:1, 2:0.5:1, 1:0.5:1.5, 1:0.5:2, 1:1:1, 1:1.5:1 or 1:2:1.

[0015] Preferably, the hydrolyzed rice extract is prepared by mixing rice with water, adding protease for hydrolysis, and then subjecting the hydrolysis solution to enzyme inactivation, filtration and concentration to obtain the hydrolyzed rice extract.

[0016] Preferably, the hydrolysis by adding protease is twice hydrolysis by adding alkaline protease and complex protease, respectively.

[0017] Preferably, the specific steps for preparing the hydrolyzed rice extract include: mixing rice residue with deionized water at a mass ratio of 1:6-10 to obtain a rice raw material solution, adding alkaline protease for hydrolysis at a temperature of 40-55℃ and adjusting the pH to 9.0-11.0, stirring for 1.5-2.5 h, inactivating the enzyme after hydrolysis, centrifuging to obtain the lower precipitate, and washing the precipitate with water; dispersing the washed precipitate in 6-10 times the mass of deionized water to obtain a precipitate dispersion solution, then adding complex protease at a temperature of 50-65℃ and adjusting the pH to 8.0-9.0, hydrolyzing for 2-3 h, then inactivating the enzyme, filtering to obtain a hydrolyzed rice filtrate, and evaporating and concentrating below 90℃ to obtain the hydrolyzed rice extract.

[0018] Preferably, the mass ratio of the alkaline protease to the rice raw material solution is 1.2:80-150.

[0019] Preferably, the mass ratio of the complex protease to the precipitated dispersion liquid is 1:200-300;

[0020] Preferably, the French chrysanthemum extract is prepared by the following method: taking the anther part of the French chrysanthemum, dispersing the pollen in water, coagulating with alcohol after filtration, drying, and then performing enzymatic hydrolysis, and then performing enzyme inactivation, filtration, and concentration to obtain the French chrysanthemum extract.

[0021] Preferably, the enzymatic hydrolysis is performed by using cellulase and α-amylase.

[0022] Preferably, the specific steps for preparing the French chrysanthemum extract include: taking the anther part of the French chrysanthemum, solubilizing the pollen in water by continuously stirring in a boiling water bath for 2-4 hours using 20-30 times the mass of deionized water, coagulating with 95% ethanol after filtration to separate water-soluble polysaccharides, and then drying, adding 3-7 times the mass of deionized water, adjusting the pH range to 4.0-5.5 and the temperature to 45-55°C to obtain a chrysanthemum raw material solution, adding cellulase and α-amylase, stirring for 4-5 hours to perform enzymatic hydrolysis, performing heat treatment to inactivate the enzyme after the enzymatic hydrolysis is completed by increasing the temperature to 70-80°C, maintaining stirring for 0.5-1 hours, then decreasing the temperature to room temperature, removing the precipitate by filtration or centrifugation, and collecting the supernatant; performing secondary filtration using an ultrafiltration membrane, and concentrating under reduced pressure to obtain the French chrysanthemum extract.

[0023] Preferably, the amount of 95% ethanol is 1-2 times the mass of the pollen after filtration;

[0024] Preferably, the mass ratio of the cellulase to the α-amylase is 1:0.5-1.5; further preferably, 1:1;

[0025] Preferably, the mass ratio of the total mass of the cellulase and the α-amylase to the chrysanthemum raw material solution is 1:200-300;

[0026] Preferably, the density of the French chrysanthemum extract is 1-1.5 g / cm 3 ; further preferably, the density of the French chrysanthemum extract is 1.1 g / cm 3 .

[0027] Preferably, the long-stem grape caulerpa extract is prepared by the following method: taking dry long-stem grape caulerpa powder, performing enzymatic hydrolysis in 8-12 times the mass of water using cellulase, performing enzyme inactivation, filtration, and concentration on the enzymatic hydrolysis solution to obtain the long-stem grape caulerpa extract.

[0028] Preferably, the mass ratio of the cellulase to the total mass of the long-stem grape caulerpa powder and water is 3:2000-5000.

[0029] Preferably, the specific steps for preparing the C. longispinosa extract include: taking dry C. longispinosa powder, adding 8-12 times the mass of deionized water, adding cellulase for enzymatic hydrolysis, the temperature is 40-60℃, the pH is adjusted to 4.5-7.0, stirring for 1-2h, after the enzymatic hydrolysis is completed, the enzyme is inactivated by heating, first filtration is performed, then the precipitate is removed by centrifugation, secondary filtration is performed using an ultrafiltration membrane, and concentration is performed to obtain the C. longispinosa extract.

[0030] Preferably, the natural composition further comprises a solvent acceptable in the cosmetic field.

[0031] Illustratively, the solvent is deionized water, caprylyl glycol, 1,2-hexanediol or 1,2-pentanediol.

[0032] Preferably, the sources of the alkaline protease, the complex protease, the cellulase and the alpha-amylase are all commercially available and are not specifically limited.

[0033] Further preferably, the alkaline protease is Alcalase 2.4L FG from Novozymes, Denmark.

[0034] Further preferably, the complex protease is a rice protein hydrolysis special compound protease produced by Donghenghuadaozhi Enzyme Preparation Company.

[0035] Further preferably, the cellulase is cellulase produced by Donghenghuadaozhi Enzyme Preparation Company.

[0036] Further preferably, the alpha-amylase is A9857 produced by Sigma-Aldrich Company.

[0037] The hydrolyzed rice extract, the French chrysanthemum extract and the C. longispinosa extract of the present application can be prepared by an enzymatic hydrolysis method, the preparation process is simple and time-saving, the storage period of the extract can be effectively shortened, and the inactivation of the active ingredients can be avoided. The extract prepared by the method of the present application not only has good anti-aging effect, but also almost no irritation, and has a wider application range.

[0038] Another aspect of the present application provides a skin care product containing the above-mentioned multiple anti-aging natural composition.

[0039] Preferably, the skin care product is an aqueous preparation, an emulsion, a cream or an eye cream.

[0040] Preferably, the raw materials of the cream include an emulsifier, a thickening agent, an emollient, a humectant, a stabilizer, a preservative and the above-mentioned multiple anti-aging natural composition.

[0041] Preferably, the raw materials of the cream include A, B, C, D, E phases, the A phase includes sucrose polystearate (and) cetyl palmitate, cetylstearyl alcohol, coco-caprylate / caprate, propylheptyl octanoate; the B phase includes water and glycerin; the C phase includes water and xanthan gum; the D phase includes undecane (and) tridecane; the E phase includes phenoxyethanol (and) methylparaben (and) ethylparaben (and) propyl glycol, the above-mentioned multiple anti-aging natural composition.

[0042] Preferably, the A phase includes sucrose polystearate (and) cetyl palmitate 1-2%, cetylstearyl alcohol 3-5%, coco-caprylate / caprate 5-10%, propylheptyl octanoate 2-5% by mass percentage of the cream; the B phase includes water 50-60% and glycerin 2-5%; the C phase includes water 15-20% and xanthan gum 0.5-2%; the D phase includes undecane (and) tridecane 1-3%; the E phase includes phenoxyethanol (and) methylparaben (and) ethylparaben (and) propyl glycol 0.5-2%, hydrolyzed rice extract (and) French helichrysum extract (and) long-stem grape caulerpa extract 1.5-3%.

[0043] Further preferably, the A phase includes sucrose polystearate (and) cetyl palmitate 1.5%, cetylstearyl alcohol 4%, coco-caprylate / caprate 8.5%, propylheptyl octanoate 2.5% by mass percentage of the cream; the B phase includes water 55.5% and glycerin 3%; the C phase includes water 19% and xanthan gum 1%; the D phase includes undecane (and) tridecane 2%; the E phase includes phenoxyethanol (and) methylparaben (and) ethylparaben (and) propyl glycol 1%, hydrolyzed rice extract (and) French helichrysum extract (and) long-stem grape caulerpa extract 2%.

[0044] Preferably, the preparation method of the cream includes the following steps: heating the A phase and the B phase to 60-85°C respectively, pre-mixing the C phase, adding the C phase to the B phase under stirring, uniformly mixing, then adding the A phase under stirring and cooling to 45-55°C, and using a homogenizer for dispersion homogenization; adding the E phase and uniformly dispersing to obtain the cream.

[0045] Preferably, the heating temperature of the A phase and the B phase is 80°C.

[0046] Preferably, the cooling temperature is 55°C.

[0047] Compared with the prior art, the present application has the following beneficial effects:

[0048] 1. The natural composition of the present application comprising hydrolyzed rice extract, French camomile extract and Caudiculae Caulerpa extract can effectively promote the synthesis of collagen, make up for the loss of collagen, and play a supporting role for the skin, and fade wrinkles;

[0049] 2. The multiple anti-aging natural composition of the present application can promote the synthesis of periostin, thereby repairing the structure of collagen fibers damaged due to aging, reducing the relaxation of the skin, and playing a skin tightening effect;

[0050] 3. The multiple anti-aging natural composition of the present application can also promote the synthesis of glycosaminoglycans, improve the water retention capacity of the skin, improve the roughness of the skin, and reduce wrinkles caused by dryness;

[0051] 4. The multiple anti-aging natural composition of the present application effectively repairs the DEJ functional interface damaged due to aging, maintains the normal function between the epidermis and the dermis, and maintains the elasticity and tightness of the skin;

[0052] 5. The multiple anti-aging natural composition of the present application cooperatively restores the skin function from multiple pathways, integrates and optimizes the matrix network, fades the existing wrinkles, delays the generation of new wrinkles, and solves the skin aging problem in multiple dimensions;

[0053] 6. The multiple anti-aging natural composition of the present application uses green and pollution-free plant extracts, conforms to the green chemistry concept, meets the market demand, and has a very high industrialization value. BRIEF DESCRIPTION OF DRAWINGS

[0054] Figure 1 Primos CR photos of the eye corner wrinkles of the volunteers before (left photo) and after 4 weeks (right photo) of using the cream of Example 7;

[0055] Figure 2 A statistical graph of the average depth of the eye corner wrinkles of the volunteers before, after 2 weeks, and after 4 weeks of using the cream of Example 7;

[0056] Figure 3 A statistical graph of the average volume of the eye corner wrinkles of the volunteers before, after 2 weeks, and after 4 weeks of using the cream of Example 7;

[0057] Figure 4 A statistical graph of the average area of the eye corner wrinkles of the volunteers before, after 2 weeks, and after 4 weeks of using the cream of Example 7. DETAILED DESCRIPTION

[0058] The enzymes in the examples are all preferred sources in the invention.

[0059] Example 1

[0060] This embodiment provides a multi-anti-aging natural composition, which is composed of hydrolyzed rice extract, French immortelle extract and long-stemmed grape fern extract, wherein the mass ratio of the hydrolyzed rice extract, French immortelle extract and long-stemmed grape fern extract is 1:0.5:1;

[0061] The specific steps of preparing the hydrolyzed rice extract are: taking rice residue and deionized water in a mass ratio of 1:7 to obtain a rice raw material liquid, adding alkaline protease for hydrolysis, the mass ratio of alkaline protease to rice raw material liquid is 1.2:100, the temperature is 55°C, the pH is adjusted to 9.0, stirring and hydrolyzing for 2 hours, heating to 80°C after the hydrolysis is completed to inactivate the enzyme, centrifuging to remove the lower layer of precipitate, and washing the precipitate three times with deionized water; dispersing the washed precipitate in 8 times the mass of deionized water to obtain a precipitate dispersion, then adding composite protease, the mass ratio of composite protease to precipitate dispersion is 1:300, the temperature is 50°C, the pH is adjusted to 9.0, hydrolyzing for 3 hours, then heating to 80°C to inactivate the enzyme, filtering to obtain a hydrolyzed rice filtrate, and evaporating and concentrating at 80°C to a density of 1.1 g / mL to obtain a hydrolyzed rice extract;

[0062] The specific steps of preparing the French immortelle extract are: taking the anther part of the French immortelle, continuously stirring for 3 hours with 20 times the mass of deionized water in a boiling water bath to solubilize the pollen in water, filtering and coagulating with an equal mass of 95% ethanol to separate water-soluble polysaccharides, then drying, adding 5 times the mass of deionized water, adjusting the pH range to 5 and the temperature to 50°C to obtain an immortelle raw material liquid, adding cellulase and α-amylase, the total mass ratio of cellulase and α-amylase to the immortelle raw material liquid is 1:300, and the mass ratio of cellulase and α-amylase is 1:1, stirring for 4 hours to perform enzymolysis, and after the enzymolysis is completed, heating to 70°C, keeping the temperature and stirring for 1 hour, heat-treating at 80°C to inactivate the enzyme, then cooling to 25°C, centrifuging to remove the precipitate, and collecting the clear liquid; secondary filtration using an ultrafiltration membrane, and concentrating under reduced pressure to a density of 1.1 g / mL to obtain the French immortelle extract;

[0063] The specific steps of preparing the long-stem grape ferns extract are: taking dried long-stem grape ferns powder, adding 10 times the mass of deionized water, adding cellulase for enzymatic hydrolysis, wherein the mass ratio of cellulase to the total mass of the long-stem grape ferns powder and water is 3:5000, the temperature is 50°C, the pH is adjusted to 6.0, and stirring is carried out for 2 hours. After the enzymatic hydrolysis is completed, the temperature is raised to 80°C to inactivate the enzyme, and a first filtration is performed, followed by centrifugation to remove the precipitate, and a second filtration is performed using an ultrafiltration membrane, and the concentration is concentrated to a density of 1.1 g / mL to obtain the long-stem grape ferns extract;

[0064] This embodiment also provides a facial cream containing the above-mentioned multiple anti-aging natural composition, including phases A, B, C, D, and E. Calculated by mass percentage of the facial cream, phase A includes 1.5% sucrose polystearate (and) cetyl palmitate, 4% cetearyl alcohol, 8.5% coconut oil caprylate / caprate, and 2.5% propylheptyl caprylate; phase B includes 55.5% water and 3% glycerin; phase C includes 19% water and 1% xanthan gum; phase D includes 2% undecane (and) tridecane; phase E includes 1% phenoxyethanol (and) methylparaben (and) ethylparaben (and) propylene glycol, and 2% of the multiple anti-aging natural composition.

[0065] The facial cream is prepared by the following steps:

[0066] Heat phase A and phase B to 80°C separately, pre-mix phase C, add phase C to phase B under stirring, mix evenly, then add phase A under stirring and cool to 55°C, use a homogenizer to disperse and homogenize; add phase E, continue to disperse evenly, and cool to room temperature (25°C).

[0067] Example 2

[0068] This embodiment provides a multi-anti-aging natural composition, which is implemented in the same manner as in Example 1, except that the mass ratio of the hydrolyzed rice extract, the French immortelle extract, and the long-stemmed grape fern extract is 1.5:0.5:1.

[0069] Example 3

[0070] This embodiment provides a multi-anti-aging natural composition, and its implementation method is the same as that of Example 1, except that the mass ratio of the hydrolyzed rice extract, the French immortelle extract, and the long-stemmed grape fern extract is 2:0.5:1.

[0071] Example 4

[0072] This embodiment provides a multi-anti-aging natural composition, which is implemented in the same manner as in Example 1, except that the mass ratio of the hydrolyzed rice extract, the French immortelle extract, and the long-stemmed grape fern extract is 1:0.5:1.5.

[0073] Example 5

[0074] This embodiment provides a multi-anti-aging natural composition, and its implementation method is the same as that of Example 1, except that the mass ratio of the hydrolyzed rice extract, the French immortelle extract, and the long-stemmed grape fern extract is 1:0.5:2.

[0075] Example 6

[0076] This embodiment provides a multiple anti-aging natural composition, and its implementation method is the same as that of Example 1, except that the mass ratio of the hydrolyzed rice extract, the French immortelle extract, and the long-stemmed grape fern extract is 1:1:1.

[0077] Example 7

[0078] This embodiment provides a multiple anti-aging natural composition and a facial cream comprising the multiple anti-aging natural composition. The implementation method is the same as that of Example 1, except that the mass ratio of the hydrolyzed rice extract, the French immortelle extract and the long-stem grape fern extract is 1:1.5:1.

[0079] Example 8

[0080] This embodiment provides a multi-anti-aging natural composition, and its implementation method is the same as that of Example 1, except that the mass ratio of the hydrolyzed rice extract, the French immortelle extract, and the long-stemmed grape fern extract is 1:2:1.

[0081] Comparative Example 1

[0082] This embodiment provides a multi-anti-aging natural composition, and its implementation method is the same as that of Example 1, except that the mass ratio of the hydrolyzed rice extract, the French immortelle extract, and the long-stemmed grape fern extract is 0:2:0.

[0083] Comparative Example 2

[0084] This embodiment provides a multi-anti-aging natural composition, and its implementation method is the same as that of Example 1, except that the mass ratio of the hydrolyzed rice extract, the French immortelle extract, and the long-stemmed grape fern extract is 1:0:1.

[0085] Performance Testing

[0086] The samples used in the performance test were the multiple anti-aging natural compositions of Examples 1 to 8 and Comparative Examples 1 to 2, which were diluted with deionized water to form sample solutions of corresponding mass concentrations.

[0087] 1. Synthesis and structural testing of the matrix network: In vitro studies were conducted on normal fibroblasts (taken from young (≤30 years old) and aged (≥50 years old) recipients) using optical microscopy and transmission electron microscopy. Fibroblasts from aged recipients were tested with a 1% concentration of the multiple anti-aging natural composition solution: young and aged human fibroblasts (donated by beauty salons) were inoculated and cultured separately at 37°C and 5% CO2. Then, 0.4 mL of a 1% concentration of the multiple anti-aging natural composition and 10 ng / mL of TGF-β (transforming growth factor β - positive control) were added to the culture medium and treated with the human fibroblasts for 7 days.

[0088] The slides were rinsed in PBS buffer (Ponsay, Wuhan).

[0089] Fixation in methanol for 4 minutes at -20°C.

[0090] Rinsing with PBS buffer (Ponsay, Wuhan).

[0091] Alexa Fluor® 488 conjugated anti-stain (Abeam) was used.

[0092] The synthesis of the collagen network was studied using visualization with an optical microscope (X70, Olympus) and an image analysis system (NIS-Elements AR, Nikon). The amount of collagen I is directly proportional to the intensity of green fluorescence. The more intense the green fluorescence, the denser the matrix network. The images were quantitatively analyzed using Matlab software to calculate the rate of increase in the synthesis of the collagen I network tested, the results of which are shown in Table 1.

[0093] Table 1

[0094]

[0095]

[0096] 2. Periostin synthesis test: ex vivo study of 3D models of young (≤ 30 years) and aged (≥ 50 years) skin explants (abdominal plastic surgery donations) by immunohistochemistry, the amount of periostin synthesis in young skin explants was 89 AU. The surface of 5 x 5 mm aged human skin explants was smeared with 0.2 mL of the Multi-Anti-Aging Natural Composition solution at a concentration of 1.5%, once a day, for 5 consecutive days, on the 6thday the explants were fixed, after dehydration they were embedded in paraffin, sections of 4 um were prepared (microtome RM2125RT, Leica), then the paraffin was stripped. Staining with Alexa Fluor® 568 fluorescent conjugate reagent, the synthesis of periostin was studied using visualization with an optical microscope (X70, Olympus) and an image analysis system (NIS-Elements AR, Nikon). The synthesis of periostin is directly proportional to the average intensity of green fluorescence (total fluorescence intensity / dermal surface). The images were quantitatively analyzed using Matlab software. The amount of periostin synthesis before and after stimulation with the Multi-Anti-Aging Natural Composition was measured and recorded in Table 2.

[0097] Table 2

[0098]

[0099] ​3. DEJ structure and function test: ex vivo study on young (< 30 years) and aged (> 50 years) skin explants 3D model by immunohistochemistry, using 0.2 mL of the multipurpose anti-aging natural composition solution at 1% concentration, applied on the surface of the aged skin explants of 5 x 5 mm, once a day, for 9 consecutive days, on the 10thday the explants were cryofrozen at a temperature of -75°C, preparing two sections of 4 um (microtome RM2125RT, Leica), one of which was stained with Propidium Iodide (Sigma-Aldrich) to show collagen IV, the other was stained with P3E4, (Santa Cruz Biotech) to show fibronectin-5. Optical observation was performed using a Leica type DMLB microscope equipped with an Olympus DP 72. camera, by fluorescent labeling, the synthesis of collagen IV and fibronectin-5 was directly proportional to the average green fluorescence intensity (total fluorescence intensity / dermal surface), comparing before and after 9 days of application, the rate of change in the synthesis of collagen IV and fibronectin-5 in the dermal-epidermal junction and in the upper papillary dermis was calculated, the results are reported in Table 3.

[0100] Table 3

[0101]

[0102] 4. Skin ECM moisturizing function test: ex vivo study on young (< 30 years) and aged (> 50 years) skin explants (abdominal plastic surgery donations) 3D model by immunohistochemistry, using 0.2 mL of the multipurpose anti-aging natural composition solution at 1% concentration, applied on the surface of the aged skin explants of 5 x 5 mm, once a day, for 9 consecutive days, GAGs-specific immunostaining was performed using the Mowry staining method (Alcian blue stain, Sigma-Aldrich). Optical observation was performed using a Leica type DMLB microscope equipped with an Olympus DP 72. camera, by staining labeling, comparing before and after 9 days of application, due to the pinkish purple staining, it was possible to see the glycosaminoglycans present in the papillary dermis layer and in the dermal-epidermal junction, observing and calculating the rate of change in the synthesis of glycosaminoglycans in the dermal-epidermal junction and in the upper papillary dermis, the results are reported in Table 4.

[0103] Table 4

[0104]

[0105]

[0106] 5. Clinical test: 15 healthy female volunteers, aged 35-60 years, were selected to continuously use the cream of Example 7 for 28 days, 0.7 mL each time in the morning and evening. Before each test, the subjects washed their faces and rested in a constant temperature and humidity room for 15 min. The Primos CR (Canfield America) was used to test the skin at the corners of the eyes of the subjects before use, after 2 weeks (14 days) of use, and after 4 weeks (28 days) of use. The Primos CR photos of the skin at the corners of the eyes of two volunteers before and after using the cream of Example 7 are shown in Figure 1 , the average depth of the wrinkles at the corners of the eyes of the volunteers before and after using the cream of Example 7 is shown in Figure 2 , the average volume of the wrinkles at the corners of the eyes of the volunteers before and after using the cream of Example 7 is shown in Figure 3 , and the average area of the wrinkles at the corners of the eyes of the volunteers before and after using the cream of Example 7 is shown in Figure 4 .

[0107] During the test, the volunteers did not have any irritation or allergic feedback, and the product did not have any delamination or caking phenomenon.

Claims

1. A multi-anti-aging natural composition, characterized in that, The hydrolyzed rice extract, the French wax chrysanthemum extract, and the caulerpa lentillifera extract; The mass ratio of the hydrolyzed rice extract, the French wax chrysanthemum extract, and the caulerpa lentillifera extract is (1-2):(0.5-1.5):(1-2); The hydrolyzed rice extract is prepared by mixing rice with water, adding protease for hydrolysis, and then performing enzyme inactivation, filtration, and concentration on the hydrolysis solution obtained by hydrolysis to obtain the hydrolyzed rice extract; The adding protease for hydrolysis is adding alkaline protease and compound protease for twice hydrolysis, respectively; The French wax chrysanthemum extract is prepared by taking the anther part of the French wax chrysanthemum, dispersing the pollen in water, condensing with alcohol after filtration, drying, and then performing enzymolysis, and then performing enzyme inactivation, filtration, and concentration on the product after enzymolysis to obtain the French wax chrysanthemum extract; The enzymolysis is performed by using cellulase and alpha-amylase for enzymolysis; The caulerpa lentillifera extract is prepared by taking dry caulerpa lentillifera powder, performing enzymolysis on the powder in 8-12 times mass of water by using cellulase, and then performing enzyme inactivation, filtration, and concentration on the enzymolysis solution to obtain the caulerpa lentillifera extract.

2. The multiplex anti-aging natural composition according to claim 1, characterized in that, The mass ratio of the cellulase to the total mass of the caulerpa lentillifera powder and water is 3:(2000-5000).

3. The multiplex anti-aging natural composition according to claim 1 or 2, characterized in that, The natural composition further comprises a solvent acceptable in the cosmetic field.

4. A skin care product comprising the multiple anti-aging natural composition according to any one of claims 1-3.

Images