Aqueous solution polymer composition as well as preparation method and application thereof

Abstract

The invention discloses an aqueous solution polymer composition as well as a preparation method and application thereof. The aqueous solution polymer composition comprises a sulfhydrylated hyaluronic acid oligosaccharide solution, rhamnose and polypeptide containing long-chain alkyl, and water is taken as a solvent. The content of hyaluronic acid in the sulfhydrylated hyaluronic acid oligosaccharide solution is 0.01 mg / mL to 20 mg / mL, the molecular weight of sulfhydrylated hyaluronic acid oligosaccharide is 1.2 kDa to 4 kDa, and the content of sulfydryl is 1 [mu] mol / g to 250 [mu] mol / g. The aqueous solution polymer composition has the effects of eliminating harmful free radicals, repairing barriers and stimulating activity enhancement of collagen, meanwhile, thiolated hyaluronic acid oligosaccharide and polypeptide containing long-chain alkyl in the aqueous solution polymer composition have a synergistic effect, the anti-aging effect is comprehensive, the effect is obvious, the duration time is long, and the anti-aging effect is good. The aqueous solution polymer composition is especially suitable for people with sensitive skin and can be applied to the field of cosmetics, and the preparation method of the aqueous solution polymer composition is simple.

Description

Technical Field

[0001] The present invention relates to the field of medical technology, and in particular to an aqueous solution polymer composition and a preparation method and application thereof. Background Art

[0002] In the normal metabolism of the human body, there is a dynamic balance between oxidation and reduction. However, under the influence of various endogenous (such as chronic or acute infection, etc.) and exogenous factors (such as environmental pollution, ultraviolet radiation, etc.), oxidation reactions may prevail, resulting in oxidative stress and the generation of various harmful free radicals. Free radicals are highly oxidizing and can cause peroxidation of unsaturated lipids in biological membranes. The formed lipid peroxides are strong cross-linking agents, which combine with proteins, nucleic acids, etc. to form insoluble substances, which can damage the body's tissues and cells, thereby causing aging effects such as dry, rough, dull, sagging, and wrinkles. The collagen in aging skin looks irregular and disorganized, the content of collagen I in the skin decreases, and the ratio of collagen III content to collagen I content increases.

[0003] The skin barrier of sensitive people is more easily damaged, and their ability to resist the harsh external environment is weaker. Seasonal changes, staying up late, air pollution and ultraviolet rays may all induce sensitive skin. At this time, the skin of sensitive skin produces more harmful free radicals, and is more prone to oxidative stress and skin aging. The anti-aging strategy for people with sensitive skin is to remove harmful free radicals, repair the barrier, and increase the content of collagen in the skin. The combination of the three will synergistically exert a three-dimensional anti-aging effect. Although there are already some technical examples of delaying skin aging, the anti-aging strategies of the prior art do not have the above three points. Therefore, it is necessary to study a composition with enhanced benefits that can remove harmful free radicals, repair the barrier, and stimulate the activity of collagen for people with sensitive skin. Summary of the invention

[0004] The technical problem to be solved by the present invention is to provide an aqueous solution polymer composition and a preparation method and application thereof. The aqueous solution polymer composition has the functions of eliminating harmful free radicals, repairing barriers, and stimulating the enhancement of collagen activity. At the same time, the thiolated hyaluronic acid oligosaccharides and the long-chain alkyl-containing polypeptides in the aqueous solution polymer composition have synergistic effects, and the anti-aging effect is comprehensive, the effect is obvious, and the duration is long. It is especially suitable for people with sensitive skin and can be applied to the field of cosmetics. The preparation method of the aqueous solution polymer composition of the present invention is simple.

[0005] In order to solve the above technical problems, the first aspect of the present invention is to provide an aqueous solution polymer composition, comprising the following components:

[0006] 0.05 to 20 parts by weight of thiolated hyaluronic acid oligosaccharide solution;

[0007] Rhamnose 0.02-5 parts by weight;

[0008] 0.001-0.2 parts by weight of a polypeptide containing a long-chain alkyl group;

[0009] The solvent is water;

[0010] The hyaluronic acid content in the thiolated hyaluronic acid oligosaccharide solution is 0.01 mg / mL to 20 mg / mL, the molecular weight of the thiolated hyaluronic acid oligosaccharide is 1.2 kDa to 4 kDa, and the thiol content of the thiolated hyaluronic acid oligosaccharide is 1 μmol / g to 250 μmol / g.

[0011] In the present invention, thiolated hyaluronic acid oligosaccharides play a role in eliminating harmful free radicals, rhamnose promotes the establishment of hemidesmosome structures at the bottom of epithelial cells, and promotes the proliferation of stratum corneum cells, thereby repairing the barrier function of the skin; long-chain alkyl-containing polypeptides can promote the generation of small molecules such as type I collagen, type III collagen, type IV collagen, hyaluronic acid, glycosaminoglycans and fibronectin in epidermal-dermal junction tissue (DEJ). Eliminating harmful free radicals can slow down the loss of collagen in the body and promote the repair of the skin barrier; and the repair of the barrier can resist the external environment, reduce the damage of oxidative stress to the body, and slow down the loss of collagen in the body; generating more collagen and fibronectin can strengthen the skin barrier and reduce inflammatory response. The combination of the three exerts a three-dimensional anti-aging effect, which is especially suitable for people with sensitive skin.

[0012] It should be noted that the transdermal absorption capacity of skin is related to the molecular weight of hyaluronic acid and its derivatives, and the lower the molecular weight, the more conducive to absorption, but too low hyaluronic acid oligosaccharides (polymerization degree 1~2) can not be recognized by receptor cells, and can not play its biological function well, therefore, in the aqueous solution polymer composition provided by the present invention, the molecular weight of thiolated hyaluronic acid oligosaccharides is preferably 1.2kDa~4kDa (polymerization degree 3-10), and the molecular weight is suitable, and it is easily absorbed by the skin and recognized by receptor cells, and can penetrate into the dermis, and has good free radical scavenging ability, and can also accelerate the proliferation of fibroblasts and the synthesis of collagen. In addition, although the thiolation content in thiolated hyaluronic acid oligosaccharides is improved, although it is conducive to eliminating harmful free radicals, too high thiol content means that more chemical modification is needed, and excessive modification may affect the recognition of hyaluronic acid by cell receptors, therefore, preferably thiol content in thiolated hyaluronic acid oligosaccharides is 1μmol / g polymer~250μmol / g polymer.

[0013] Specifically, the thiolated hyaluronic acid oligosaccharide solution comprises 0.05 to 20 parts by mass or 2.0 to 10 parts by mass, with 2.0 to 10 being preferred parts by mass; rhamnose comprises 0.02 to 5 parts by mass or 0.1 to 2 parts by mass, with 0.1 to 2 being preferred parts by mass; a polypeptide containing a long-chain alkyl group comprises 0.001 to 0.2 parts by mass or 0.005 to 0.05 parts by mass, with 0.005 to 0.05 being preferred parts by mass; and a solvent comprises 50 to 150 parts by mass.

[0014] Specifically, the polypeptide containing a long-chain alkyl group is a polypeptide containing a C16 long-chain alkyl group; wherein the polypeptide is selected from one or more of a tripeptide, a pentapeptide, and a hexapeptide, for example, the polypeptide is preferably one or more of palmitoyl tripeptide-1, palmitoyl tripeptide-5, palmitoyl tripeptide-8, palmitoyl pentapeptide-4, palmitoyl pentapeptide-5, palmitoyl hexapeptide-12, palmitoyl hexapeptide-14, and palmitoyl hexapeptide-15, and the polypeptide is more preferably palmitoyl tripeptide-1 and / or palmitoyl tripeptide-5. In this specific embodiment, the polypeptide is chemically modified, such as carrying a long-chain alkyl group, which can enhance the skin affinity of the molecule, especially the polypeptide acylated with palmitic acid of a C16 long-chain alkyl group can significantly improve skin permeability, biological activity and economic feasibility.

[0015] Specifically, the mass ratio between the thiolated hyaluronic acid oligosaccharide solution and the long-chain alkyl-containing polypeptide is 2:1-100:1 or 5:1-20:1, wherein 5:1-20:1 is the preferred mass ratio.

[0016] Specifically, the hyaluronic acid content in the thiolated hyaluronic acid oligosaccharide solution is 0.1 mg / mL to 10 mg / mL.

[0017] In order to solve the above technical problems, the second aspect of the present invention is to provide an application of the aforementioned aqueous solution polymer composition in the preparation of anti-aging cosmetics. Due to the aqueous solution polymer composition provided by the present invention, its main structure is easily recognized by cell receptors, and the active thiol groups carried by it can remove harmful free radicals, and its polypeptide containing long-chain alkyl can promote fibroblasts to produce collagen, thereby improving the ability of cells to recognize thiol-containing hyaluronic acid oligosaccharides, and can remove more harmful free radicals, and promote the anti-aging effect of thiol-containing hyaluronic acid oligosaccharides, therefore, it can be used in the preparation of anti-aging cosmetics. For example, the aqueous solution polymer composition provided by the present invention is combined with auxiliary materials and / or other active ingredients such as moisturizers, soothing agents, skin conditioners, antioxidants, thickeners, emulsifiers, preservatives, acid-base regulators, chelating agents, etc. to obtain anti-aging cosmetics. In the present invention, anti-aging cosmetics can be, for example, facial masks, essence water, essence, lotion, face cream, eye cream, lip essence, body milk, and shower gel.

[0018] In order to solve the above technical problems, the third aspect of the present invention is to provide a method for preparing the aforementioned aqueous solution polymer composition, comprising the following steps:

[0019] Step 1, using sodium hyaluronate as a raw material, subjecting it to thiol modification to obtain a thiol-modified hyaluronic acid solution having a thiol content of 1 μmol / g to 250 μmol / g;

[0020] Step 2, placing the thiolated hyaluronic acid solution obtained in step 1 into a dialysis bag with a molecular weight cutoff of 1.2 kDa for dialysis, and collecting the feed liquid in the dialysis bag;

[0021] Step 3, filtering the liquid collected in step 2 through an ultrafiltration membrane with a molecular weight cutoff of 4.0 kDa, collecting the filtrate, and obtaining a thiolated hyaluronic acid oligosaccharide solution with a molecular weight of 1.2 kDa to 4 kDa;

[0022] Step 4: Add purified water to the thiolated hyaluronic acid oligosaccharide solution of step 3 to adjust the content of hyaluronic acid to 0.01 mg / mL to 20 mg / mL; mix the thiolated hyaluronic acid oligosaccharide solution, rhamnose and a polypeptide containing a long-chain alkyl group, and add solvent water to obtain an aqueous solution polymer composition.

[0023] Specifically, in step 4, the content of hyaluronic acid in the thiolated hyaluronic acid oligosaccharide solution of step 3 is adjusted to 0.1 mg / mL to 10 mg / mL; the thiolated hyaluronic acid oligosaccharide solution is 0.05 to 20 parts by mass or 0.2 to 5 parts by mass, 0.2 to 5 being preferred parts by mass; rhamnose is 0.02 to 5 parts by mass or 0.1 to 2 parts by mass, 0.1 to 2 being preferred parts by mass; the long-chain alkyl-containing polypeptide is 0.001 to 0.2 parts by mass or 0.005 to 0.05 parts by mass, 0.005 to 0.05 being preferred parts by mass; and the solvent is 50 to 100 parts by mass.

[0024] Specifically, in the polymer composition solution obtained in step 4, the mass ratio between the thiolated hyaluronic acid oligosaccharide solution and the polypeptide containing a long-chain alkyl group is 2:1 to 100:1; more specifically, in the polymer composition solution obtained in step 4, the mass ratio between the thiolated hyaluronic acid oligosaccharide solution and the polypeptide containing a long-chain alkyl group is 5:1 to 20:1 or 5:1 to 20:1, wherein 5:1 to 20:1 is a preferred mass ratio.

[0025] Specifically, the solvent water in step 4 is purified water.

[0026] The preparation method provided by the present invention is simple to operate and easy to industrialize. It should be noted that in the present invention, step 1 is a method of using sodium hyaluronate as a raw material and subjecting it to thiol modification, for example, the method disclosed by Shu et al. (Shu et al., Biomacromolecules, 2002, 3: 1304-1311) can be adopted, specifically, the raw material hyaluronic acid is dissolved in water, hydrochloric acid is added for acid hydrolysis, and under the action of a catalyst carbodiimide and an activating reagent N-hydroxysulfosuccinimide, it reacts with a diamine containing a disulfide bond, and finally a reducing agent is added to reduce the disulfide bond to a free thiol group to obtain a thiol-modified hyaluronic acid solution. BRIEF DESCRIPTION OF THE DRAWINGS

[0027] In order to more clearly illustrate the technical solution of the present invention, the following briefly introduces the drawings required for use in the present invention. Obviously, the drawings described below are only some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying creative work.

[0028] Figure 1 This is the result of human efficacy test on the anti-aging product prepared in Example 1. DETAILED DESCRIPTION

[0029] The following will be combined with the accompanying drawings to clearly and completely describe the technical solutions in the present invention. Obviously, the described embodiments are part of the embodiments of the present invention, rather than all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0030] Example 1

[0031] Step 1: 1.0g of sodium hyaluronate (MW 10kDa) was dissolved in 100mL of distilled water, 3mL of 6mol / L hydrochloric acid was added, and the solution was acidified at 40°C for 20 minutes. An appropriate amount of acid or base (0.1N hydrochloric acid or sodium hydroxide) was added to adjust the solution to a specific pH value of 4.75. 0.12g of Sulfo-NHS and 1.2g of dithiodipropionic acid dihydrazide (DTP) were added, and then 0.54g of EDCI solid was added, stirred and dissolved, and an appropriate amount of acid or base (0.1N hydrochloric acid or sodium hydroxide) was added to keep the pH value of the reaction solution stable at the aforementioned specific value, and the reaction was stirred overnight. 1.0N sodium hydroxide was added to adjust the pH value of the reaction solution to 8.5 to terminate the reaction. Then 0.5g of dithiothreitol was added, stirred and dissolved, and the reaction was allowed to react for 4 hours. Then 1.0N hydrochloric acid was added to adjust the pH value of the above reaction solution to 3.0 to obtain a thiolated hyaluronic acid solution;

[0032] Step 2: The thiolated hyaluronic acid solution obtained in step 1 is placed into a dialysis bag with a molecular weight cutoff of 1.2 kDa for dialysis, and the feed liquid in the dialysis bag is collected;

[0033] Step 3: Pass the liquid collected in step 2 through an ultrafiltration membrane with a molecular weight cutoff of 4.0 kDa, collect the permeate, and obtain a thiolated hyaluronic acid oligosaccharide solution. The total hyaluronic acid content is 8.02 mg / mL and the thiol content is 51.5 μmol / g polymer.

[0034] Step 4: Add purified water to adjust the total hyaluronic acid content in the thiolated hyaluronic acid oligosaccharide solution in step 3 to 6.0 mg / mL, take 5 g of the thiolated hyaluronic acid oligosaccharide solution, add 1.0 g of rhamnose and 0.025 g of palmitoyl tripeptide-1, and stir evenly;

[0035] Step 5: Add 0.5 g of p-hydroxyacetophenone and 0.5 g of 1,2-hexanediol to the product prepared in step 4, adjust the amount of purified water to make the total mixed solution 100 g, stir evenly, and the anti-aging cosmetic is obtained; similarly, repeat steps 4 and 5 to prepare anti-aging cosmetics of any mass.

[0036] Example 2

[0037] Step 1: 1.0 g of sodium hyaluronate (MW 10 kDa) was dissolved in 100 mL of distilled water, 3 mL of 6 mol / L hydrochloric acid was added, and the solution was acidified at 40 ° C for 20 minutes. An appropriate amount of acid or base (0.1 N hydrochloric acid or sodium hydroxide) was added to adjust the solution to a specific pH value of 4.75. 0.12 g of Sulfo-NHS and 1.2 g of dithiodipropionic acid dihydrazide (DTP) were added, and then 0.54 g of EDCI solid was added, stirred and dissolved, and an appropriate amount of acid or base (0.1 N hydrochloric acid or sodium hydroxide) was added to keep the pH value of the reaction solution stable at the aforementioned specific value, and the reaction was stirred overnight. 1.0 N sodium hydroxide was added to adjust the pH of the reaction solution to 8.5 to terminate the reaction. Then 0.5 g of dithiothreitol was added, stirred and dissolved, and the reaction was allowed to react for 4 hours. Then 1.0 N hydrochloric acid was added to adjust the pH value of the above reaction solution to 3.0 to obtain a thiolated hyaluronic acid solution;

[0038] Step 2: The thiolated hyaluronic acid solution obtained in step 1 is placed into a dialysis bag with a molecular weight cutoff of 1.2 kDa for dialysis, and the feed liquid in the dialysis bag is collected;

[0039] Step 3: The liquid collected in step 2 is passed through an ultrafiltration membrane with a molecular weight cutoff of 4.0 kDa, and the permeate is collected, which is the thiolated hyaluronic acid oligosaccharide solution. The total hyaluronic acid content is 8.02 mg / mL and the thiol content is 51.5 μmol / g polymer.

[0040] Step 4: Add purified water to adjust the total hyaluronic acid content in the thiolated hyaluronic acid oligosaccharide solution in step 3 to 6.0 mg / mL, take 5 g of the thiolated hyaluronic acid oligosaccharide solution, add 1.0 g of rhamnose and 0.025 g of palmitoyl tripeptide-5, and stir evenly;

[0041] Step 5: Add 0.5 g of p-hydroxyacetophenone and 0.5 g of 1,2-hexanediol to the product prepared in step 4, adjust the amount of purified water to make the total mixed solution 100 g, stir evenly, and the anti-aging cosmetic is obtained; similarly, repeat steps 4 and 5 to prepare anti-aging cosmetics of any mass.

[0042] Example 3

[0043] Step 1: 1.0 g of sodium hyaluronate (MW 10 kDa) was dissolved in 100 mL of distilled water, 3 mL of 6 mol / L hydrochloric acid was added, and the solution was acidified at 40 ° C for 20 minutes. An appropriate amount of acid or base (0.1 N hydrochloric acid or sodium hydroxide) was added to adjust the solution to a specific pH value of 4.75. 0.12 g of Sulfo-NHS and 1.2 g of dithiodipropionic acid dihydrazide (DTP) were added, and then 0.36 g of EDCI solid was added, stirred and dissolved, and an appropriate amount of acid or base (0.1 N hydrochloric acid or sodium hydroxide) was added to keep the pH value of the reaction solution stable at the aforementioned specific value, and the reaction was stirred overnight. 1.0 N sodium hydroxide was added to adjust the pH value of the reaction solution to 8.5 to terminate the reaction. Then 0.5 g of dithiothreitol was added, stirred and dissolved, and the reaction was allowed to react for 4 hours. Then 1.0 N hydrochloric acid was added to adjust the pH value of the above reaction solution to 3.0 to obtain a thiolated hyaluronic acid solution;

[0044] Step 2: The thiolated hyaluronic acid solution obtained in step 1 is placed into a dialysis bag with a molecular weight cutoff of 1.2 kDa for dialysis, and the feed liquid in the dialysis bag is collected;

[0045] Step 3: The liquid collected in step 2 is passed through an ultrafiltration membrane with a molecular weight cutoff of 4.0 kDa, and the permeate is collected, which is the thiolated hyaluronic acid oligosaccharide solution. The total hyaluronic acid content is 8.05 mg / mL and the thiol content is 25.7 μmol / g polymer.

[0046] Step 4: Add purified water to adjust the total hyaluronic acid content in the thiolated hyaluronic acid oligosaccharide solution in step 3 to 6.0 mg / mL, take 5 g of the thiolated hyaluronic acid oligosaccharide solution, add 1.0 g of rhamnose and 0.025 g of palmitoyl tripeptide-1, and stir evenly;

[0047] Step 5: Add 0.5 g of p-hydroxyacetophenone and 0.5 g of 1,2-hexanediol to the product prepared in step 4, adjust the amount of purified water to make the total mixed solution 100 g, stir evenly, and the anti-aging cosmetic is obtained; similarly, repeat steps 4 and 5 to prepare anti-aging cosmetics of any mass.

[0048] Example 4

[0049] Step 1: 1.0 g of sodium hyaluronate (MW 10 kDa) was dissolved in 100 mL of distilled water, 3 mL of 6 mol / L hydrochloric acid was added, and the solution was acidified at 40 ° C for 20 minutes. An appropriate amount of acid or base (0.1 N hydrochloric acid or sodium hydroxide) was added to adjust the solution to a specific pH value of 4.75. 0.12 g of Sulfo-NHS and 1.2 g of dithiodipropionic acid dihydrazide (DTP) were added, and then 0.72 g of EDCI solid was added, stirred and dissolved, and an appropriate amount of acid or base (0.1 N hydrochloric acid or sodium hydroxide) was added to keep the pH value of the reaction solution stable at the aforementioned specific value, and the reaction was stirred overnight. 1.0 N sodium hydroxide was added to adjust the pH value of the reaction solution to 8.5 to terminate the reaction. Then 0.5 g of dithiothreitol was added, stirred and dissolved, and the reaction was allowed to react for 4 hours. Then 1.0 N hydrochloric acid was added to adjust the pH value of the above reaction solution to 3.0 to obtain a thiolated hyaluronic acid solution;

[0050] Step 2: The thiolated hyaluronic acid solution obtained in step 1 is placed into a dialysis bag with a molecular weight cutoff of 1.2 kDa for dialysis, and the feed liquid in the dialysis bag is collected;

[0051] Step 3: The liquid collected in step 2 is passed through an ultrafiltration membrane with a molecular weight cutoff of 4.0 kDa, and the permeate is collected, which is the thiolated hyaluronic acid oligosaccharide solution. The total hyaluronic acid content is 8.08 mg / mL and the thiol content is 99.3 μmol / g polymer.

[0052] Step 4: Add purified water to adjust the total hyaluronic acid content in the thiolated hyaluronic acid oligosaccharide solution in step 3 to 6.0 mg / mL, take 5 g of the thiolated hyaluronic acid oligosaccharide solution, add 1.0 g of rhamnose and 0.025 g of palmitoyl tripeptide-1, and stir evenly;

[0053] Step 5: Add 0.5 g of p-hydroxyacetophenone and 0.5 g of 1,2-hexanediol to the product prepared in step 4, adjust the amount of purified water to make the total mixed solution 100 g, stir evenly, and the anti-aging cosmetic is obtained; similarly, repeat steps 4 and 5 to prepare anti-aging cosmetics of any mass.

[0054] Embodiments 5 to 14

[0055] Examples 5 to 14: The preparation method is the same as that of Example 1, except that the amount of thiolated hyaluronic acid oligosaccharide solution, rhamnose and palmitoyl tripeptide-1 added to the aqueous solution polymer composition is different. The thiol content (51.5 μmol / g polymer), p-hydroxyacetophenone (0.5 g), and 1,2-hexanediol (0.5 g) added remain unchanged, and the amount of purified water is adjusted to make the total mixed solution 100 g. The amount of thiolated hyaluronic acid oligosaccharide solution, rhamnose and palmitoyl tripeptide-1 added in Examples 5 to 14 is detailed in Table 1; similarly, in Examples 5 to 14, by scaling up or down in the same proportion, any mass of anti-aging cosmetics can be prepared.

[0056] Table 1

[0057] Example Thiolated hyaluronic acid oligosaccharide solution / g Rhamnose / g Palmitoyl tripeptide-1 / g Example 5 5.0 1.0 0.005 Example 6 5.0 1.0 0.05 Example 7 2 0.1 0.005 Example 8 2 0.1 0.05 Example 9 2 2.0 0.005 Example 10 2 2.0 0.05 Embodiment 11 10.0 0.1 0.005 Example 12 10.0 0.1 0.05 Example 13 10.0 2.0 0.005 Embodiment 14 10.0 2.0 0.05

[0058] Embodiments 15-16

[0059] Examples 15-16: The preparation method is the same as that of Example 1, except that in the composition, the amount of thiolated hyaluronic acid oligosaccharide solution is controlled to be 2.5 g, the amount of rhamnose is controlled to be 1.0 g, and the amount of palmitoyl tripeptide-1 added is changed. The thiol content (51.5 μmol / g polymer), the amount of p-hydroxyacetophenone (0.5 g), and the amount of 1,2-hexanediol (0.5 g) added are all unchanged, and the amount of purified water is adjusted to make the total mixed solution 100 g. The amount of thiolated hyaluronic acid oligosaccharide solution, rhamnose, and palmitoyl tripeptide-1 added in Examples 15-16 is detailed in Table 2; in Examples 15-16, by scaling up or down in the same proportion, anti-aging cosmetics of any quality can be prepared.

[0060] Table 2

[0061] Example Thiolated hyaluronic acid oligosaccharide solution / g Rhamnose / g Palmitoyl tripeptide-1 / g Embodiment 15 2.5 1.0 0.2 Example 16 2.5 1.0 0.05

[0062] Comparative Example 1

[0063] Step 1: 0.6g hyaluronic acid (molecular weight 1.2kDa ~ 4.0kDa), add purified water to 100g, stir until dissolved, take 2.5g of the above hyaluronic acid oligosaccharide solution, add 1.0g rhamnose and 0.025g palmitoyl tripeptide-1, and stir evenly;

[0064] Step 2: Add 0.5 g of p-hydroxyacetophenone and 0.5 g of 1,2-hexanediol to the product prepared in step 4, adjust the amount of purified water to make the total mixed solution 100 g, stir evenly, and the cosmetic reference substance is obtained; by scaling up or down in the same proportion, any mass of cosmetic reference substance can be prepared.

[0065] Comparative Example 2

[0066] Step 1: 1.0 g of sodium hyaluronate (MW 10 kDa) was dissolved in 100 mL of distilled water, 3 mL of 6 mol / L hydrochloric acid was added, and the solution was acidified at 40 ° C for 20 minutes. An appropriate amount of acid or base (0.1 N hydrochloric acid or sodium hydroxide) was added to adjust the solution to a specific pH value of 4.75. 0.12 g of Sulfo-NHS and 1.2 g of dithiodipropionic acid dihydrazide (DTP) were added, and then 0.72 g of EDCI solid was added, stirred and dissolved, and an appropriate amount of acid or base (0.1 N hydrochloric acid or sodium hydroxide) was added to keep the pH value of the reaction solution stable at the aforementioned specific value, and the reaction was stirred overnight. 1.0 N sodium hydroxide was added to adjust the pH of the reaction solution to 8.5 to terminate the reaction. Then 0.5 g of dithiothreitol was added, stirred and dissolved, and the reaction was allowed to react for 4 hours. Then 1.0 N hydrochloric acid was added to adjust the pH value of the above reaction solution to 3.0 to obtain a thiolated hyaluronic acid solution;

[0067] Step 2: The thiolated hyaluronic acid solution obtained in step 1 is placed into a dialysis bag with a molecular weight cutoff of 1.2 kDa for dialysis, and the feed liquid in the dialysis bag is collected;

[0068] Step 3: The liquid collected in step 2 is passed through an ultrafiltration membrane with a molecular weight cutoff of 4.0 kDa, and the permeate is collected, which is the thiolated hyaluronic acid oligosaccharide solution. The total hyaluronic acid content is 10.02 mg / mL and the thiol content is 75.5 μmol / g polymer.

[0069] Step 4: Add purified water to adjust the total hyaluronic acid content in the thiolated hyaluronic acid oligosaccharide solution in step 3 to 6.0 mg / mL, and take 5 g of the thiolated hyaluronic acid oligosaccharide solution for standby use;

[0070] Step 5: Add 0.5 g of p-hydroxyacetophenone and 0.5 g of 1,2-hexanediol to 5 g of the thiolated hyaluronic acid oligosaccharide solution in step 4, adjust the amount of purified water to make the total mixed solution 100 g, and stir evenly to obtain the cosmetic reference substance. By scaling up or down in the same proportion, any mass of cosmetic reference substance can be prepared.

[0071] Comparative Example 3

[0072] Step 1: Add 1.0g of rhamnose, 0.5g of p-hydroxyacetophenone and 0.5g of 1,2-hexanediol, adjust the amount of purified water to make the total mixed solution 100g, stir evenly, and the cosmetic reference substance is obtained; by scaling up or down in the same proportion, any mass of cosmetic reference substance can be prepared.

[0073] Comparative Example 4

[0074] Step 1: Add 0.025g of palmitoyl tripeptide-1, 0.5g of p-hydroxyacetophenone and 0.5g of 1,2-hexanediol, adjust the amount of purified water to make the total mixed solution 100g, stir evenly, and the cosmetic reference substance is obtained; by enlarging or reducing the same proportion, any mass of cosmetic reference substance can be prepared.

[0075] Comparative Example 5

[0076] Step 1: 1.0 g of sodium hyaluronate (MW 10 kDa) was dissolved in 100 mL of distilled water, 3 mL of 6 mol / L hydrochloric acid was added, and the solution was acidified at 40 ° C for 20 minutes. An appropriate amount of acid or base (0.1 N hydrochloric acid or sodium hydroxide) was added to adjust the solution to a specific pH value of 4.75. 0.12 g of Sulfo-NHS and 1.2 g of dithiodipropionic acid dihydrazide (DTP) were added, and then 0.72 g of EDCI solid was added, stirred and dissolved, and an appropriate amount of acid or base (0.1 N hydrochloric acid or sodium hydroxide) was added to keep the pH value of the reaction solution stable at the aforementioned specific value, and the reaction was stirred overnight. 1.0 N sodium hydroxide was added to adjust the pH of the reaction solution to 8.5 to terminate the reaction. Then 0.5 g of dithiothreitol was added, stirred and dissolved, and the reaction was allowed to react for 4 hours. Then 1.0 N hydrochloric acid was added to adjust the pH value of the above reaction solution to 3.0 to obtain a thiolated hyaluronic acid solution;

[0077] Step 2: The thiolated hyaluronic acid solution obtained in step 1 is placed into a dialysis bag with a molecular weight cutoff of 1.2 kDa for dialysis, and the feed liquid in the dialysis bag is collected;

[0078] Step 3: The liquid collected in step 2 is passed through an ultrafiltration membrane with a molecular weight cutoff of 4.0 kDa, and the permeate is collected, which is the thiolated hyaluronic acid oligosaccharide solution. The total hyaluronic acid content is 10.02 mg / mL and the thiol content is 75.5 μmol / g polymer.

[0079] Step 4: Add purified water to adjust the total hyaluronic acid content in the thiolated hyaluronic acid oligosaccharide solution in step 3 to 6.0 mg / mL, take 5 g of the above thiolated hyaluronic acid oligosaccharide solution, add 1.0 g of rhamnose, and stir evenly;

[0080] Step 5: Add 0.5 g of p-hydroxyacetophenone and 0.5 g of 1,2-hexanediol to the product prepared in step 4, adjust the amount of purified water to make the total mixed solution 100 g, and stir evenly to obtain the cosmetic reference substance. By scaling up or down in the same proportion, any mass of cosmetic reference substance can be prepared.

[0081] Comparative Example 6

[0082] Step 1: Add 1.0g of rhamnose, 0.025g of palmitoyl tripeptide-1, 0.5g of p-hydroxyacetophenone and 0.5g of 1,2-hexanediol, adjust the amount of purified water to make the total mixed solution 100g, stir evenly, and the cosmetic reference substance is obtained; by scaling up or down in the same proportion, any mass of cosmetic reference substance can be prepared.

[0083] Test Example 1 Firming and Anti-wrinkle Efficacy Test - In vitro Keratinocyte Reactive Oxygen Species (ROS) Inhibition Test Method

[0084] Test principle and test method: 2',7'-Dichlorodi-hydrofluorescein diacetate (DCFH-DA) has no spontaneous fluorescence and can freely pass through the cell membrane. After entering the cell, it is hydrolyzed by esterase to generate DCFH, which cannot penetrate the cell membrane and accumulates in the cell. The reactive oxygen species in the cell can oxidize the non-fluorescent DCFH to generate fluorescent DCF. The green fluorescence intensity is proportional to the level of reactive oxygen in the cell. Hydrogen peroxide (H2O2), ultraviolet rays, blue light, etc., as exogenous oxidants, can induce an increase in the ROS content in human keratinocytes. The ROS inhibition rate is calculated by comparing the changes in the fluorescence intensity of the intracellular fluorescent dye DCFH-DA after administration of the negative control and the test substance.

[0085] The calculation formula of ROS inhibition rate is:

[0086]

[0087] T—the average of three fluorescence intensities of the tested sample;

[0088] C—average of three times of fluorescence intensity of negative control;

[0089] C0—the average of three fluorescence intensities of the naked cell group.

[0090] The samples of Examples 1 to 16, Comparative Examples 1 to 2, and Comparative Examples 5 to 6 were diluted with PBS buffer to a thiolated hyaluronic acid oligosaccharide / hyaluronic acid oligosaccharide content of 0.1 mg / mL. Comparative Examples 3 to 4 were not diluted. The test results are shown in Table 3.

[0091] Table 3

[0092]

[0093]

[0094] From the test results in the above table, it can be seen that the inhibition rate values ​​of Examples 1 to 16 on human epidermal keratinocytes are all greater than 80% (SD), and all have the function of inhibiting ROS and the inhibitory effect. The results are more than 10% higher than those of Comparative Examples 2 and 5, and more than 20% higher than those of Comparative Example 1.

[0095] Test Example 2 Human skin patch test

[0096] Test sample: Anti-aging product of Example 1.

[0097] Negative control: distilled water.

[0098] Test method: Refer to the "Technical Specifications for Safety of Cosmetics" (2015 edition) and recruit 30 volunteers who meet the requirements. Use a patch test device with an area of ​​no more than 50mm2 and a depth of about 1mm. Add the test substance to the patch test device, with an amount of about 0.020mL. Stick the patch test sample with the test substance and the negative control on the curved side of the subject's forearm, and remove the patch test device after 24 hours. Observe the skin reaction according to the standard 30 minutes after removal (after the pressure disappears), 24 hours and 48 hours, and record the observation results. The evaluation criteria are shown in Table 4, and the evaluation results are shown in Table 5.

[0099] Table 4

[0100]

[0101] Table 5

[0102]

[0103] The test results show that the 30 subjects scored 0 at 0.5h, 24h, and 48h after removing the test article spot tester, and no skin reaction occurred. This shows that the anti-aging product prepared in Example 1 is mild, non-irritating to the human body, and extremely safe.

[0104] Test Example 3 Human Efficacy Test

[0105] Test sample: the anti-aging product prepared in Example 1;

[0106] Test environment: temperature 21±1℃, humidity 50±10%RH;

[0107] Test site: face;

[0108] Subjects: 30 subjects with sensitive skin (lactic acid stinging ≥ 3 points), aged 25-55 years;

[0109] Test instruments: Tewameter TM Hex (water loss test probe TEWL), Reviscometer RVM600 (elastic fiber tissue test probe), skin ultrasound diagnostic instrument Ultrascan UC-22.

[0110] Test the basic value of the subject's skin: the subject cleans the face, wipes the skin with dry tissue, and sits in the test environment for 30 minutes. Laboratory technicians use Tewameter TM Hex to test the transepidermal water loss rate of the skin, ReviscometerRVM600 to test the skin elasticity, and UC-22 to test the skin dermal density.

[0111] Apply 1.0 mL of cosmetics evenly on the subject's face, once in the morning and once in the evening;

[0112] Test time: 0 days, 14 days, 28 days.

[0113] The improvement rate of transepidermal water loss (%) was calculated according to the following formula:

[0114] Improvement rate of transepidermal water loss (%) = (1-measurement value after application / measurement value before application) × 100%

[0115] The greater the improvement rate (%) of skin transepidermal water loss, the better the moisturizing effect.

[0116] The skin elasticity improvement rate (%) was calculated according to the following formula:

[0117] Skin elasticity improvement rate (%) = (1-measurement value after application / measurement value before application) × 100%

[0118] The greater the skin elasticity improvement rate (%), the better the repair effect.

[0119] The percentage increase in skin dermis density (%) was calculated according to the following formula:

[0120] Increased percentage of skin dermis density (%) = (1-measured value after application / measured value before application) × 100%

[0121] The greater the percentage increase (%) of skin dermis density, the better the repair effect.

[0122] The test results can be found in Figure 1 .

[0123] Test 3 The anti-aging product prepared in Example 1 was tested for efficacy on 30 people with sensitive skin, and the test results showed excellent effects. By 28 days, the improvement rate of transepidermal water loss was 36.9%, the improvement rate of skin elasticity was 31.4%, and the increase percentage of dermal density was 29.1%, indicating that the anti-aging product prepared in Example 1 has excellent anti-aging efficacy.

[0124] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. It should be pointed out that a person skilled in the art can make several improvements and modifications without departing from the technical principles of the present invention, and these improvements and modifications should also be regarded as within the scope of protection of the present invention.

Claims

1. An aqueous solution polymer composition, characterized in that The components include: 0.05 to 20 parts by weight of thiolated hyaluronic acid oligosaccharide solution; Rhamnose 0.02-5 parts by weight; 0.001-0.2 parts by weight of a polypeptide containing a long-chain alkyl group; The solvent is water; The hyaluronic acid content in the thiolated hyaluronic acid oligosaccharide solution is 0.01 mg / mL to 20 mg / mL, the molecular weight of the thiolated hyaluronic acid oligosaccharide is 1.2 kDa to 4 kDa, and the thiol content of the thiolated hyaluronic acid oligosaccharide is 1 μmol / g to 250 μmol / g.

2. The aqueous solution polymer composition according to claim 1, characterized in that The thiolated hyaluronic acid oligosaccharide solution comprises 0.05 to 20 parts by mass or 2.0 to 10 parts by mass, rhamnose comprises 0.02 to 5 parts by mass or 0.1 to 2 parts by mass, a polypeptide containing a long-chain alkyl group comprises 0.001 to 0.2 parts by mass or 0.005 to 0.05 parts by mass, and a solvent comprises 50 to 150 parts by mass.

3. The aqueous solution polymer composition according to claim 1, characterized in that The long-chain alkyl-containing polypeptide is a C16 long-chain alkyl polypeptide; Wherein, the polypeptide is selected from one or more of tripeptide, pentapeptide and hexapeptide.

4. The aqueous solution polymer composition according to claim 1, characterized in that The mass ratio between the thiolated hyaluronic acid oligosaccharide solution and the long-chain alkyl-containing polypeptide is 2:1 to 100:1 or 5:1 to 20:

1.

5. The aqueous solution polymer composition according to claim 1, characterized in that: The hyaluronic acid content in the thiolated hyaluronic acid oligosaccharide solution is 0.1 mg / mL to 10 mg / mL.

6. Use of the aqueous solution polymer composition according to any one of claims 1 to 5 in the preparation of anti-aging cosmetics.

7. A method for preparing an aqueous solution polymer composition according to any one of claims 1 to 5, characterized in that: The steps include: Step 1, using sodium hyaluronate as a raw material, subjecting it to thiol modification to obtain a thiol-modified hyaluronic acid solution having a thiol content of 1 μmol / g to 250 μmol / g; Step 2, placing the thiolated hyaluronic acid solution obtained in step 1 into a dialysis bag with a molecular weight cutoff of 1.2 kDa for dialysis, and collecting the feed liquid in the dialysis bag; Step 3, filtering the liquid collected in step 2 through an ultrafiltration membrane with a molecular weight cutoff of 4.0 kDa, collecting the filtrate, and obtaining a thiolated hyaluronic acid oligosaccharide solution with a molecular weight of 1.2 kDa to 4 kDa; Step 4: Add purified water to the thiolated hyaluronic acid oligosaccharide solution of step 3 to adjust the content of hyaluronic acid to 0.01 mg / mL to 20 mg / mL; mix the thiolated hyaluronic acid oligosaccharide solution, rhamnose and a polypeptide containing a long-chain alkyl group, and add solvent water to obtain an aqueous solution polymer composition.

8. The preparation method according to claim 7, characterized in that: In step 4, the hyaluronic acid content in the thiolated hyaluronic acid oligosaccharide solution of step 3 is adjusted to 0.1 mg / mL to 10 mg / mL; the thiolated hyaluronic acid oligosaccharide solution is 0.05 to 20 parts by mass or 0.2 to 5 parts by mass, rhamnose is 0.02 to 5 parts by mass or 0.1 to 2 parts by mass, the long-chain alkyl-containing polypeptide is 0.001 to 0.2 parts by mass or 0.005 to 0.05 parts by mass, and the solvent is 50 to 100 parts by mass.

9. The preparation method according to claim 7, characterized in that: In the polymer composition solution obtained in step 4, the mass ratio between the thiolated hyaluronic acid oligosaccharide solution and the long-chain alkyl-containing polypeptide is 2:1 to 100:1 or 5:1 to 20:

1.

10. The preparation method according to claim 7, characterized in that: The solvent water in step 4 is purified water.

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