Composition for reducing degradation capacity of hyaluronidase on hyaluronic acid or salt thereof and application of composition

Abstract

The invention provides a composition containing hyaluronic acid or salt thereof. The composition comprises hyaluronic acid or salt thereof, collagen substances and chondroitin sulfate. Degradation of the hyaluronic acid or the salt thereof is inhibited and / or slowed down, the effect of the hyaluronic acid or the salt thereof is maintained, the safety risk of a human body caused by addition of an enzyme preparation in a formula is avoided, and the effect of the hyaluronic acid or the salt thereof is safely and effectively prolonged.

Description

[0001] This application is a divisional application of Chinese patent application No. 202310467886X, filed on April 24, 2023, entitled "A use, method and composition for inhibiting and / or slowing down the decomposition of hyaluronic acid or its salts, and / or maintaining the effects of hyaluronic acid or its salts". The contents of the parent application are incorporated herein by reference. Technical Field

[0002] The present application relates to the field of cosmetic technology, and in particular to a composition for reducing the ability of hyaluronidase to degrade hyaluronic acid or its salts and its use. Background Art

[0003] Hyaluronic acid (HA) is widely used in skincare formulations as a natural moisturizing factor. However, once applied to the skin, it is degraded into oligosaccharides or monosaccharides by the skin's HA-degrading enzymes, losing its intended skincare benefits. Existing technologies have been used to slow HA degradation by adding enzyme inhibitors to formulas, but this poses a safety risk and may lead to HA metabolic diseases. Summary of the Invention

[0004] To solve the above problems, the inventors of the present application discovered that collagen substances of a certain molecular weight, or a combination of collagen substances of a certain molecular weight and chondroitin sulfate, can inhibit and / or slow down the decomposition of hyaluronic acid or its salts, maintain the effects of hyaluronic acid or its salts, and thus complete the present invention.

[0005] The technical solution of this application is as follows:

[0006] 1. Use of a collagen substance, a combination of a collagen substance and chondroitin sulfate, a composition comprising a collagen substance, or a composition comprising a collagen substance and chondroitin sulfate for inhibiting and / or slowing the decomposition of hyaluronic acid or a salt thereof, and / or maintaining the effect of hyaluronic acid or a salt thereof.

[0007] 2. The use according to item 1, wherein the molecular weight of the collagen-like substance is 1w-9w Da.

[0008] 3. The use according to item 1, wherein

[0009] When the collagen substance is used to achieve the above purpose, the mass ratio of the collagen substance to the hyaluronic acid or its salt is (0.5-200):1; or,

[0010] When the combination of the collagen substance and chondroitin sulfate is used to achieve the above purpose, the mass ratio of the sum of the mass of the chondroitin sulfate and the collagen substance to the mass of the hyaluronic acid or its salt is (0.5-200):1. Preferably, the mass ratio of the chondroitin sulfate and the collagen substance is greater than or equal to 1:1, and more preferably is (1-29):1.

[0011] 4. The use according to item 1, wherein the molecular weight of the hyaluronic acid or its salt used to achieve the use is 180w Da or less.

[0012] 5. A composition containing hyaluronic acid or a salt thereof, wherein the composition comprises hyaluronic acid or a salt thereof and a collagen-like substance, and the molecular weight of the collagen-like substance is 1w-9w Da;

[0013] Preferably, the mass ratio of the collagen substance to the hyaluronic acid or its salt is (0.5-200):1.

[0014] 6. A composition containing hyaluronic acid or a salt thereof, wherein the composition comprises hyaluronic acid or a salt thereof, a collagen substance and chondroitin sulfate;

[0015] Preferably, the molecular weight of the collagen-like substance is 1w-9w Da;

[0016] Preferably, the mass ratio of the sum of the mass of the chondroitin sulfate and the collagen substance to the mass of the hyaluronic acid or its salt is (0.5-200):1;

[0017] Preferably, the mass ratio of the chondroitin sulfate to the collagen substance is greater than or equal to 1:1, more preferably (1-29):1.

[0018] 7. The composition according to item 5 or 6, wherein

[0019] The molecular weight of the hyaluronic acid or its salt is less than 180w Da.

[0020] 8. Use of the composition according to any one of items 5 to 7 in cosmetics.

[0021] 9. Use of the composition according to any one of items 5 to 7 for inhibiting and / or slowing down the decomposition of hyaluronic acid or a salt thereof, and / or maintaining the effect of hyaluronic acid or a salt thereof.

[0022] 10. A method for inhibiting and / or slowing down the decomposition of hyaluronic acid or its salt, and / or maintaining the effect of hyaluronic acid or its salt, comprising the following steps:

[0023] A collagen-based substance, or a combination of a collagen-based substance and chondroitin sulfate, is added to an aqueous medium containing hyaluronic acid or a salt thereof.

[0024] 11. The method according to item 10, wherein the molecular weight of the collagen-like substance is 1w-9w Da;

[0025] Preferably, the mass ratio of the collagen substance to the hyaluronic acid or its salt, or the mass ratio of the combination of the collagen substance and chondroitin sulfate to the hyaluronic acid or its salt is (0.5-200):1;

[0026] Preferably, the mass ratio of the chondroitin sulfate to the collagen substance is greater than or equal to 1:1, more preferably (1-29):1;

[0027] Preferably, the molecular weight of the hyaluronic acid or its salt is less than 180w Da.

[0028] Compared with the prior art, the beneficial effects of the present application are: inhibiting and / or slowing down the degradation of hyaluronic acid or its salts, maintaining the effect of hyaluronic acid or its salts, avoiding the human safety risks brought about by adding enzyme preparations to the formula, and safely and effectively prolonging the effect of hyaluronic acid or its salts. DETAILED DESCRIPTION

[0029] The present application is further described below with reference to examples. It should be understood that the examples are only used to further illustrate and explain the present application and are not intended to limit the present application.

[0030] Unless otherwise defined, technical and scientific terms used in this specification have the same meaning as those commonly understood by those skilled in the art. Although methods and materials similar or identical to those described herein may be used in experiments or practical applications, the materials and methods are described herein below. In the event of a conflict, the present specification, including definitions, will prevail. In addition, the materials, methods, and examples are provided for illustrative purposes only and are not intended to be limiting. The present application is further described below with reference to specific examples, which are not intended to limit the scope of this application.

[0031] The present application provides the use of collagen substances, combinations of collagen substances and chondroitin sulfate, compositions comprising collagen substances, or compositions comprising collagen substances and chondroitin sulfate in inhibiting and / or slowing the decomposition of hyaluronic acid or its salts, and / or maintaining the effect of hyaluronic acid or its salts; that is, the present application provides the use of collagen substances in inhibiting and / or slowing the decomposition of hyaluronic acid or its salts, or the use of a combination of collagen substances and chondroitin sulfate in inhibiting and / or slowing the decomposition of hyaluronic acid or its salts, or the use of collagen substances in maintaining the effect of hyaluronic acid or its salts, or the use of a combination of collagen substances and chondroitin sulfate in maintaining the effect of hyaluronic acid or its salts.

[0032] In some embodiments of the present application, the type of the collagen-like substance is not limited in this application, as long as the molecular weight can be 1w-9wDa. For example, it can be collagen obtained by hydrolysis or collagen that has not been hydrolyzed.

[0033] In some embodiments of the present application, the molecular weight of the collagen-like substance is 1w-9wDa; preferably, the molecular weight of the collagen-like substance is 3w-7w Da; for example, the molecular weight of the collagen-like substance can be 1wDa, 2wDa, 3wDa, 4wDa, 5wDa, 6wDa, 7wDa, 8wDa, 9wDa or any range therebetween.

[0034] In some embodiments of the present application, the collagen-like substance includes hydrolyzed collagen.

[0035] The hydrolyzed collagen, i.e., the hydrolyzate of collagen, is a product obtained by hydrolyzing collagen. This application does not impose any restrictions on the hydrolysis method, and for example, an alkaline method, an acid method, a saline-alkali method, and / or an enzymatic method may be used. This application also does not impose any restrictions on the source of the collagen, and for example, the collagen may be extracted from animal skin, tendon, or bone tissue.

[0036] In some embodiments of the present application, the hydrolyzed collagen includes gelatin.

[0037] In some embodiments of the present application, the collagen-like substance includes soluble collagen.

[0038] The soluble collagen is soluble collagen from tissues, and can be generally divided into neutral salt soluble collagen and acid soluble collagen according to the different soluble solvents used.

[0039] In some embodiments of the present application, when the collagen substance is used to achieve the purpose, the mass ratio of the collagen substance to the hyaluronic acid or its salt is (0.5-200):1; for example, it can be 0.5:1, 1:1, 2:1, 3:1, 4:1, 5:1, 6:1, 7:1, 8:1, 9:1, 10:1, 15:1, 20:1, 25:1, 30:1, 3 1, 170:1, 180:1, 190:1, 200:1 or any range therebetween.

[0040] In some embodiments of the present application, when the combination of the collagen substance and chondroitin sulfate is used to achieve the purpose, the mass ratio of the sum of the mass of the chondroitin sulfate and the collagen substance to the mass of the hyaluronic acid or its salt is (0.5-200):1; for example, it can be 0.5:1, 1:1, 2:1, 3:1, 4:1, 5:1, 6:1, 7:1, 8:1, 9:1, 10:1, 15:1, 20:1, 1, 25:1, 30:1, 35:1, 40:1, 45:1, 50:1, 55:1, 60:1, 65:1, 70:1, 75:1, 80:1, 85:1, 90:1, 95:1, 100:1, 110:1, 120:1, 130:1, 140:1, 150:1, 160:1, 170:1, 180:1, 190:1, 200:1 or any range therebetween.

[0041] In some embodiments of the present application, the salt of hyaluronic acid is sodium salt, potassium salt, magnesium salt, calcium salt, zinc salt, bismuth salt of hyaluronic acid, or a combination of two or more thereof.

[0042] In some embodiments of the present application, the molecular weight of the hyaluronic acid or its salt is less than 180w Da, for example, it can be 180w Da, 170w Da, 160w Da, 150w Da, 140w Da, 130w Da, 120w Da, 110w Da, 100w Da, 90w Da, 80w Da, 70w Da, 60w Da, 50w Da, 40w Da, 30w Da, 20w Da, 10w Da, 9w Da, 8wDa, 7w Da, 6wDa, 5w Da, 4w Da, 3w Da, 2w Da, 1w Da, 9000Da, 8000Da, 7000Da, 6000Da, 5000Da, 4000Da, 3000Da, 2000Da, 1000Da, less than 1000Da or any range therebetween.

[0043] In some embodiments of the present application, when the combination of the collagen substance and chondroitin sulfate is used to achieve the described purpose, the mass ratio of the chondroitin sulfate to the collagen substance is greater than or equal to 1:1, preferably (1-29):1; the mass ratio of the chondroitin sulfate to the collagen substance can be 1:1, 2:1, 3:1, 4:1, 5:1, 6:1, 7:1, 8:1, 9:1, 10:1, 11:1, 12:1, 13:1, 14:1, 15:1, 16:1, 17:1, 18:1, 19:1, 20:1, 21:1, 22:1, 23:1, 24:1, 25:1, 26:1, 27:1, 28:1, 29:1 or greater than 29:1, or any range therebetween.

[0044] In some embodiments of the present application, when the collagen substance is used to achieve the purpose, the mass ratio of the collagen substance to the hyaluronic acid or its salt is (0.5~200):1, wherein the molecular weight of the collagen substance is 1w-9w Da, and the molecular weight of the hyaluronic acid or its salt is less than 180w Da.

[0045] In some embodiments of the present application, when the collagen substance is used to achieve the purpose, the mass ratio of the collagen substance to the hyaluronic acid or its salt is (0.5~200):1, wherein the molecular weight of the collagen substance is 3w-7w Da, and the molecular weight of the hyaluronic acid or its salt is less than 180w Da.

[0046] In some embodiments of the present application, when the combination of the collagen substance and chondroitin sulfate is used to achieve the purpose, the mass ratio of the sum of the masses of the chondroitin sulfate and the collagen substance to the mass of the hyaluronic acid or its salt is (0.5-200):1, and the mass ratio of the chondroitin sulfate and the collagen substance is greater than or equal to 1:1, wherein the molecular weight of the collagen substance is 1w-9w Da.

[0047] In some embodiments of the present application, when the combination of the collagen substance and chondroitin sulfate is used to achieve the purpose, the mass ratio of the sum of the masses of the chondroitin sulfate and the collagen substance to the hyaluronic acid or its salt is (0.5~200):1, and the mass ratio of the chondroitin sulfate and the collagen substance is greater than or equal to 1:1, wherein the molecular weight of the collagen substance is 1w-9w Da, and the molecular weight of the hyaluronic acid or its salt is less than 180w Da.

[0048] In some embodiments of the present application, when the combination of the collagen substance and chondroitin sulfate is used to achieve the described purpose, the mass ratio of the sum of the masses of the chondroitin sulfate and the collagen substance to the hyaluronic acid or its salt is (0.5~200):1, and the mass ratio of the chondroitin sulfate and the collagen substance is greater than or equal to 1:1, wherein the molecular weight of the collagen substance is 3w-7w Da, and the molecular weight of the hyaluronic acid or its salt is less than 180w Da.

[0049] In some embodiments of the present application, when the combination of the collagen substance and chondroitin sulfate is used to achieve the purpose, the mass ratio of the sum of the masses of the chondroitin sulfate and the collagen substance to the mass of the hyaluronic acid or its salt is (0.5-200):1, and the mass ratio of the chondroitin sulfate to the collagen substance is (1-29):1, wherein the molecular weight of the collagen substance is 1w-9w Da.

[0050] In some embodiments of the present application, when the combination of the collagen substance and chondroitin sulfate is used to achieve the described purpose, the mass ratio of the sum of the masses of the chondroitin sulfate and the collagen substance to the hyaluronic acid or its salt is (0.5-200):1, and the mass ratio of the chondroitin sulfate and the collagen substance is (1-29):1, wherein the molecular weight of the collagen substance is 1w-9w Da, and the molecular weight of the hyaluronic acid or its salt that can achieve the described purpose is less than 180w Da.

[0051] In some embodiments of the present application, when the combination of the collagen substance and chondroitin sulfate is used to achieve the purpose, the mass ratio of the sum of the masses of the chondroitin sulfate and the collagen substance to the hyaluronic acid or its salt is (0.5-200):1, and the mass ratio of the chondroitin sulfate and the collagen substance is (1-29):1, wherein the molecular weight of the collagen substance is 3w-7w Da, and the molecular weight of the hyaluronic acid or its salt is less than 180w Da.

[0052] The present application provides a composition containing hyaluronic acid or its salt, wherein the composition comprises the above-mentioned hyaluronic acid or its salt and the above-mentioned collagen-like substance; or, the composition comprises a combination of the above-mentioned hyaluronic acid or its salt, the above-mentioned collagen-like substance and the above-mentioned chondroitin sulfate.

[0053] In some embodiments of the present application, when the composition includes the above-mentioned hyaluronic acid or its salt and the above-mentioned collagen-like substance, the mass ratio of the collagen-like substance to the hyaluronic acid or its salt is (0.5~200):1, wherein the molecular weight of the collagen-like substance is 1w-9w Da, and the molecular weight of the hyaluronic acid or its salt is less than 180w Da.

[0054] In some embodiments of the present application, when the composition includes the above-mentioned hyaluronic acid or its salt and the above-mentioned collagen-like substance, the mass ratio of the collagen-like substance to the hyaluronic acid or its salt is (0.5~200):1, wherein the molecular weight of the collagen-like substance is 3w-7w Da, and the molecular weight of the hyaluronic acid or its salt is less than 180w Da.

[0055] In some embodiments of the present application, when the composition includes a combination of the above-mentioned hyaluronic acid or its salt, the above-mentioned collagen substance and the above-mentioned chondroitin sulfate, the mass ratio of the sum of the masses of the chondroitin sulfate and the collagen substance to the mass of the hyaluronic acid or its salt is (0.5~200):1, and the mass ratio of the chondroitin sulfate and the collagen substance is greater than or equal to 1:1, wherein the molecular weight of the collagen substance is 1w-9w Da, and the molecular weight of the hyaluronic acid or its salt is less than 180w Da.

[0056] In some embodiments of the present application, when the composition includes a combination of the above-mentioned hyaluronic acid or its salt, the above-mentioned collagen substance and the above-mentioned chondroitin sulfate, the mass ratio of the sum of the masses of the chondroitin sulfate and the collagen substance to the mass of the hyaluronic acid or its salt is (0.5~200):1, and the mass ratio of the chondroitin sulfate and the collagen substance is greater than or equal to 1:1, wherein the molecular weight of the collagen substance is 3w-7w Da, and the molecular weight of the hyaluronic acid or its salt is less than 180w Da.

[0057] In some embodiments of the present application, when the composition includes a combination of the above-mentioned hyaluronic acid or its salt, the above-mentioned collagen substance and the above-mentioned chondroitin sulfate, the mass ratio of the sum of the masses of the chondroitin sulfate and the collagen substance to the mass of the hyaluronic acid or its salt is (0.5-200):1, and the mass ratio of the chondroitin sulfate to the collagen substance is (1-29):1, wherein the molecular weight of the collagen substance is 1w-9w Da.

[0058] In some embodiments of the present application, when the composition includes a combination of the above-mentioned hyaluronic acid or its salt, the above-mentioned collagen substance and the above-mentioned chondroitin sulfate, the mass ratio of the sum of the masses of the chondroitin sulfate and the collagen substance to the mass of the hyaluronic acid or its salt is (0.5-200):1, and the mass ratio of the chondroitin sulfate to the collagen substance is (1-29):1, wherein the molecular weight of the collagen substance is 1w-9w Da, and the molecular weight of the hyaluronic acid or its salt is less than 180w Da.

[0059] In some embodiments of the present application, when the composition includes a combination of the above-mentioned hyaluronic acid or its salt, the above-mentioned collagen substance and the above-mentioned chondroitin sulfate, the mass ratio of the sum of the masses of the chondroitin sulfate and the collagen substance to the mass of the hyaluronic acid or its salt is (0.5-200):1, and the mass ratio of the chondroitin sulfate to the collagen substance is (1-29):1, wherein the molecular weight of the collagen substance is 3w-7w Da, and the molecular weight of the hyaluronic acid or its salt is less than 180w Da.

[0060] Regarding the above-mentioned collagen substances, the mass ratio of the above-mentioned collagen substances to the above-mentioned hyaluronic acid or its salt, the mass ratio of the sum of the mass of the above-mentioned chondroitin sulfate and the above-mentioned collagen substances to the above-mentioned hyaluronic acid or its salt, and the mass ratio of hyaluronic acid or its salt, chondroitin sulfate and the collagen substances, please refer to the above and will not be repeated here.

[0061] The present application provides use of the above composition in cosmetics.

[0062] The present application provides the use of the above composition in inhibiting and / or slowing down the decomposition of hyaluronic acid or its salt, and / or maintaining the effect of hyaluronic acid or its salt.

[0063] The present application provides a method for inhibiting and / or slowing down the decomposition of hyaluronic acid or its salt, and / or maintaining the effect of hyaluronic acid or its salt, the method comprising the following steps:

[0064] The collagen-based substance, or a combination of the collagen-based substance and the chondroitin sulfate, is added to an aqueous medium containing the hyaluronic acid or a salt thereof.

[0065] The aqueous medium may contain not only water but also other ingredients suitable for use in cosmetics.

[0066] Regarding the above-mentioned collagen substances, the mass ratio of the above-mentioned collagen substances to the above-mentioned hyaluronic acid or its salt, the mass ratio of the sum of the mass of the above-mentioned chondroitin sulfate and the above-mentioned collagen substances to the above-mentioned hyaluronic acid or its salt, and the mass ratio of hyaluronic acid or its salt, chondroitin sulfate and the collagen substances, please refer to the above and will not be repeated here.

[0067] In the present application, there is no limitation on the preparation method of the composition, which can be prepared according to conventional methods in the art.

[0068] Example

[0069] Table 1 Materials and equipment information

[0070]

[0071] CS: Chondroitin sulfate; Col(A): Collagen hydrolysate (A); Col(B): Collagen hydrolysate (B); Col(C): Collagen hydrolysate (C); Col(D): Collagen hydrolysate (D); HA: Hyaluronic acid

[0072] Examples 1-14, Comparative Examples 1-9

[0073] Examples 1-14 and Comparative Examples 1-9 were mixed according to the composition described in Table 2, with the remainder being water, to obtain an aqueous mixture solution.

[0074] Experimental example

[0075] Take the prepared sterilized surface dish and use a pipette to transfer the mixture (0.2mL / cm 2 ) Add the material to the center and shake the watch glass to spread the material evenly on the surface (*Note: The material can be self-leveling and does not require the help of a glass short stick);

[0076] After the spreading is basically completed, place the surface dish in an oven to dry (37°C, RH 20%, 30 min) until it is basically dry;

[0077] Take bacterial hyaluronidase and prepare enzyme solution using enzymatic medium (sodium hydrogen phosphate-sodium dihydrogen phosphate-sodium chloride buffer solution with 0.01% calcium chloride added, pH 6.5);

[0078] Use a pipette to add 150 μL of enzyme solution to the essentially dried surface dish containing the sample, spread it evenly, and then place the surface dish and the sample in it in a transparent sealed bag;

[0079] The samples were placed in a constant temperature oven (37°C) for a specified time (2 h, 4 h, 6 h, etc.);

[0080] After the time is up, take it out and collect the sample for leaching;

[0081] The absorbance of the collected liquid was measured at 232 nm using an ultraviolet spectrophotometer;

[0082] When the absorbance obtained by the test is stable, the first time point at which the absorbance stabilizes is considered the end point of HA degradation. At the same time, the corresponding control group without HA is used as a blank control, and the absorbance at the corresponding complete degradation and the absorbance at the test time point are recorded;

[0083] The absorbance at each time point (At) / the absorbance at the end point of HA degradation (Ae) is the HA degradation rate at each time point;

[0084] Note: The HA degradation rate detection method, the specific configuration of the enzyme solution, and the absorbance detection are all carried out in accordance with the description of the patent CN 109238995 A published by Bloomage Biotechnology.

[0085] Parameter definition:

[0086] HA degradation rate = At / Ae×100%

[0087] Where At is the absorbance value at each time point, and Ae is the absorbance value at the end point of enzymatic hydrolysis.

[0088] The results of each embodiment and comparative example are shown in Table 2.

[0089] Table 2

[0090]

[0091]

[0092] Although the present invention has been disclosed above with reference to the embodiments, they are not intended to limit the present invention. Anyone with ordinary knowledge in the art may make slight changes and modifications without departing from the spirit and scope of the present invention. Therefore, the scope of protection of the present invention shall be determined by the scope of the patent application attached hereto.

Claims

1. A composition for reducing the ability of hyaluronidase to degrade hyaluronic acid or its salts, wherein: The composition comprises the hyaluronic acid or its salt, collagen-like substance and chondroitin sulfate; The molecular weight of the collagen substance is 1w-9w Da.

2. The composition according to claim 1, wherein The mass ratio of the sum of the mass of the chondroitin sulfate and the collagen substance to the mass of the hyaluronic acid or its salt is (0.5-200):

1.

3. The composition according to claim 1, wherein The mass ratio of the chondroitin sulfate to the collagen substance is greater than or equal to 1:

1.

4. The composition according to claim 3, wherein The mass ratio of the chondroitin sulfate to the collagen substance is (1-29):

1.

5. The composition according to claim 1, wherein The molecular weight of the hyaluronic acid or its salt is less than 180w Da.

6. Use of the composition according to any one of claims 1 to 5 in cosmetics.

7. Use of the composition according to any one of claims 1 to 5 for inhibiting and / or slowing down the decomposition of hyaluronic acid or its salt, and / or maintaining the effect of hyaluronic acid or its salt.

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