A system for increasing the penetration rate and extending the duration of action of active ingredients in cosmetics or medicines

Abstract

The present invention provides a composition for improving the permeability of active ingredients of drugs or cosmetics, characterized in that the composition comprises a component 1 for increasing the water content of keratin in the stratum corneum of the skin, and a component 2 for binding the active ingredient to the keratin in the skin through ionic bonds and / or covalent bonds. The composition prolongs the action time of the active ingredient of the drug or cosmetic. The present invention also provides the use of the composition in improving the permeability of the active ingredient of the drug or cosmetic. When component 2 is prepared from a polypeptide or keratin polypeptide with an amino acid sequence such as that shown in SEQ ID NO: 1, the penetration-promoting effect on the active ingredient of the drug or cosmetic is particularly significant, achieving unexpected technical effects.

Description

Technical Field

[0001] The invention relates to the fields of medicine and cosmetics, and in particular to a system for improving the permeability of active ingredients of medicines or cosmetics and prolonging the action time. Background Art

[0002] The skin has a strong barrier function, which is a challenge for the effective delivery of exogenous active ingredients. Active ingredients such as some peptides usually have poor permeability, and most peptides must penetrate deep into the skin to exert their effects. Topical administration of drugs to the skin or topical application of cosmetics often cannot meet the concentration requirements of active ingredients.

[0003] There is still an urgent need to find methods to enhance the skin penetration of active ingredients.

[0004] The patent with announcement number CN105131086B discloses a polypeptide GMCCSR that can be used in the fields of biopharmaceuticals and cosmetics. The polypeptide can resist oxidation, protect red blood cells from hemolysis, and promote the proliferation of human skin fibroblasts and collagen production. Summary of the invention

[0005] The inventors have found through in-depth research that by using a specific ingredient to increase the water content of keratin in the stratum corneum of the skin, and then dissolving the active ingredient in a polythiol compound (non-limiting example, keratin polypeptide) solution and applying it to the skin, the active ingredient can be stably released and penetrate into the skin, thereby increasing the penetration rate of the active ingredient of the drug or cosmetic and prolonging the action time.

[0006] In a specific embodiment of the present invention, the allantoin-glycerol-hyaluronic acid mixture is first applied to the skin; the active ingredient (especially the polypeptide GMCCSR which can be used in the fields of biopharmaceuticals and cosmetics, from patent CN105131086B, which can resist oxidation, protect red blood cells from hemolysis, and promote the proliferation of human skin fibroblasts and collagen production) is dissolved in an aqueous solution containing a polythiol compound (especially the polypeptide SCP-01 [SEQ ID NO: 1] or a hydrolyzed keratin polypeptide), and then applied to the skin, a large amount of active ingredients can be stably released and penetrate into the skin.

[0007] Without being bound by theory, after the allantoin-glycerol-hyaluronic acid mixed system (component 1) acts on the skin, the water content of skin keratin increases, for example, it is higher than 20% of the water content of keratin after daily facial washing; after the active ingredient is dissolved in the aqueous solution containing the polythiol compound, the two form a disulfide bond and covalently bond (component 2). Under the condition that the water content of skin keratin increases and swells slightly, component 2 is added, and component 2 undergoes a thiol-dithiol exchange reaction with the disulfide bond of skin keratin through the thiol group in the polythiol compound (for example, the thiol group of free cysteine ​​in polypeptides), thereby forming a covalent bond coupling. As the water in the stratum corneum evaporates, the aforementioned covalent bonding is stably present in the stratum corneum. However, during daily washing, sweating, and oiling, it is difficult to achieve a state of keratin swelling, and the active ingredient is not easy to be washed away, and the concentration can be gradually increased with daily use. For example, when component 2 is applied for the second time, the thiol in component 2 continues to undergo a thiol-dithiol exchange reaction with the active ingredient to release the active ingredient; as the skin environment changes, the active ingredient gradually dissociates, and after reaching a certain concentration, it enters the subcutaneous tissue with the help of solubility difference. And the barrier function of the skin may have undergone slight, local changes. Because of the existence of the skin's self-protection mechanism, this diffusion process will not result in local burst release. In addition, keratinocytes are in a gradual metabolic process and are completely renewed approximately every 28 days. With daily use, a certain concentration of active ingredients is gradually enriched in the stratum corneum, but due to the daily renewal of keratinocytes, some active ingredients will also be metabolized, and will not significantly damage the barrier function of the stratum corneum, avoiding the impact on the overall microenvironment of the skin.

[0008] Therefore, in a first aspect of the present invention, a composition for improving the permeability or permeability of active ingredients of drugs and / or cosmetics is provided, characterized in that the composition comprises: component 1 and component 2.

[0009] The present invention provides a composition for improving the permeability of active ingredients of medicines or cosmetics, characterized in that the composition comprises: component 1 and component 2.

[0010] The present invention provides a composition for improving the permeability of active ingredients of a medicine, characterized in that the composition comprises: component 1 and component 2.

[0011] The present invention provides a composition for improving the permeability of active ingredients in cosmetics, characterized in that the composition comprises: component 1 and component 2.

[0012] In some embodiments of the present invention, the component 1 is used to increase the water content of keratin in the stratum corneum of the skin. In some embodiments of the present invention, the component 2 is used to bind the active ingredient to the keratin in the skin through ionic bonds and / or covalent bonds.

[0013] The present invention provides a composition for improving the permeability of active ingredients of medicines or cosmetics, characterized in that the composition comprises:

[0014] (1) component 1, used to increase the water content of keratin in the stratum corneum of the skin; and

[0015] (2) Component 2, used to bind the active ingredient to the skin keratin via ionic bonds and / or covalent bonds.

[0016] The present invention provides a composition for improving the permeability of an active ingredient of a drug, characterized in that the composition comprises:

[0017] (1) component 1, used to increase the water content of keratin in the stratum corneum of the skin; and

[0018] (2) Component 2, used to bind the active ingredient to the skin keratin via ionic bonds and / or covalent bonds.

[0019] The present invention provides a composition for improving the permeability of active ingredients in cosmetics, characterized in that the composition comprises:

[0020] (1) component 1, used to increase the water content of keratin in the stratum corneum of the skin; and

[0021] (2) Component 2, used to bind the active ingredient to the skin keratin via ionic bonds and / or covalent bonds.

[0022] In some embodiments of the present invention, the composition prolongs the duration of action of the active ingredient of the drug or cosmetic.

[0023] In some embodiments of the present invention, the drug or cosmetic is applied to the skin surface. In some embodiments of the present invention, the drug is applied to the skin surface. In some embodiments of the present invention, the cosmetic is applied to the skin surface.

[0024] In some embodiments of the present invention, the component 1 comprises one, two or more selected from allantoin, hyaluronic acid, vitamin E, squalane, glycerin, sodium lactate, urea, seaweed extract, aloe extract, ceramide, and glycosaminoglycan.

[0025] Preferably, the component 1 comprises allantoin, glycerol and glycosaminoglycan. Optionally, the mass ratio of allantoin, glycerol and glycosaminoglycan is 55:35:8.

[0026] In some embodiments of the present invention, the component 1 comprises allantoin, glycerol, glycosaminoglycan, and auxiliary materials.

[0027] In some embodiments of the present invention, the glycosaminoglycan comprises one, two or more selected from hyaluronic acid, chondroitin, dermatan, heparin, heparan, and keratan.

[0028] In some embodiments of the invention, the glycosaminoglycan is hyaluronic acid.

[0029] In some embodiments of the present invention, the component 1 comprises allantoin, glycerol, hyaluronic acid. In some embodiments of the present invention, the component 1 comprises allantoin, glycerol, hyaluronic acid, and excipients.

[0030] In some embodiments of the present invention, the component 1 comprises 55% by weight of allantoin, 35% by weight of glycerin, 8% by weight of hyaluronic acid, and 2% by weight of hydroxypropyl methylcellulose.

[0031] In some embodiments of the present invention, the hyaluronic acid is from Sigma-Aldrich (Product Number 924474). In some embodiments of the present invention, the allantoin is from Supelco (PHR1364).

[0032] In some embodiments of the present invention, the component 1 comprises 55% allantoin, 35% glycerol, 8% D-trehalose, and 2% hydroxypropyl methylcellulose in mass proportions.

[0033] In some embodiments of the present invention, the component 1 is prepared by the following steps: allantoin, glycerol, glycosaminoglycan and auxiliary materials are mixed and stirred to obtain. In some embodiments of the present invention, the component 1 is prepared by the following steps: 55% allantoin, 35% glycerol, 8% hyaluronic acid and 2% hydroxypropyl methylcellulose are mixed in a mass ratio, and stirred at 100r / min for 1h to obtain a paste.

[0034] In some embodiments of the present invention, the component 1 is prepared according to the method of Example 2.

[0035] In some embodiments of the present invention, the component 1 is a paste.

[0036] In some embodiments of the present invention, the component 2 comprises a keratin polypeptide or a polypeptide having an amino acid sequence as shown in SEQ ID NO: 1.

[0037] In some embodiments of the present invention, the component 2 comprises keratin polypeptides. In some embodiments of the present invention, the component 2 comprises hydrolyzed keratin polypeptides. In some embodiments of the present invention, the component 2 comprises Discoverls hydrolyzed keratin polypeptides. In some embodiments of the present invention, the component 2 comprises Puriactives hydrolyzed keratin BioActive HK20. In some embodiments of the present invention, the component 2 comprises dimercaptopropanol.

[0038] In some embodiments of the present invention, the component 2 comprises polypeptide SCP-01, and the amino acid sequence of the polypeptide SCP-01 is shown in SEQ ID NO: 1, namely LFCAGAPNSWDILRGCKVDSTTIYQHG.

[0039] In some embodiments of the present invention, the component 2 comprises a polypeptide having an amino acid sequence as shown in SEQ ID NO:1.

[0040] In some embodiments of the present invention, component 2 is prepared by the following steps: dissolving the active ingredient in a keratin polypeptide and / or silk peptide solution, adding oligoarginine and / or lysine, stirring, dialyzing and / or ultrafiltering the reaction solution, adjusting the pH value, adding zinc lactate, and freeze-drying.

[0041] In some embodiments of the present invention, the component 2 is prepared by the following steps: dissolving the active ingredient in a keratin polypeptide solution, adding oligoarginine, stirring, ultrafiltering the reaction solution, adjusting the pH value to 5.5, adding zinc lactate, and freeze-drying.

[0042] In some embodiments of the present invention, the component 2 is prepared by the following steps: dissolving 1 g of a drug or cosmetic active ingredient in 500 mL of an aqueous solution of 3 g of a polythiol compound, adding 40 mg of poly-L-arginine, and stirring at room temperature for 5 h; ultrafiltering the reaction solution using an NMWL 30 kDa ultrafiltration membrane, adjusting the pH value to 5.5 using dilute hydrochloric acid, adding 20 mg of zinc lactate, and freeze-drying to obtain the product.

[0043] In some embodiments of the present invention, the active ingredient is the polypeptide GMCCSR from patent CN105131086B, or the polypeptide AHK-Cu. In some embodiments of the present invention, the active ingredient is the polypeptide GMCCSR (SEQ ID NO: 2) from patent CN105131086B. In some embodiments of the present invention, the active ingredient is the polypeptide AHK-Cu.

[0044] In some embodiments of the present invention, the active ingredient is the polypeptide AHK-Cu (No. 411881) from Peptide Biotechnology.

[0045] In some embodiments of the present invention, the polythiol compound is selected from a polypeptide having an amino acid sequence as shown in SEQ ID NO: 1, a hydrolyzed keratin polypeptide or dimercaptopropanol.

[0046] In some embodiments of the present invention, the polythiol compound is a polypeptide with an amino acid sequence as shown in SEQ ID NO: 1 (named as polypeptide SCP-01). In some embodiments of the present invention, the polythiol compound is a hydrolyzed keratin polypeptide. In some embodiments of the present invention, the polythiol compound is dithiopropanol. In some embodiments of the present invention, the polythiol compound is Discoverls hydrolyzed keratin polypeptide. In some embodiments of the present invention, the polythiol compound is Puriactives hydrolyzed keratin BioActive HK20.

[0047] In some embodiments of the present invention, the poly-L-arginine is from Sigma-Aldrich (P4663, molwt5000-15000).

[0048] In some embodiments of the present invention, the component 2 is prepared by the following steps: dissolving 1 g of a polypeptide having an amino acid sequence as shown in SEQ ID NO: 2 in 500 mL of an aqueous solution of 3 g of a polypeptide having an amino acid sequence as shown in SEQ ID NO: 1, adding 40 mg of poly-L-arginine, and stirring at room temperature for 5 hours; ultrafiltration of the reaction solution using an NMWL 30 kDa ultrafiltration membrane, adjusting the pH value to 5.5 using dilute hydrochloric acid, adding 20 mg of zinc lactate, and freeze-drying to obtain the product.

[0049] In some embodiments of the present invention, the component 2 is prepared according to the method of Example 2.

[0050] In some embodiments of the present invention, the component 2 is optionally selected from component 2a, component 2b, component 2c, component 2d, component 2e, component 2f, component 2g, component 2h, component 2m, and component 2n prepared according to the method of Example 2.

[0051] In some embodiments of the present invention, the component 2 is selected from component 2a, component 2b, component 2c, component 2d, component 2e, component 2f, component 2g, and component 2h prepared according to the method of Example 2.

[0052] In some embodiments of the present invention, the component 2 is the component 2a prepared according to the method of Example 2. In some embodiments of the present invention, the component 2 is the component 2b prepared according to the method of Example 2.

[0053] The present invention provides a composition for improving the permeability of active ingredients of medicines or cosmetics, characterized in that the composition comprises:

[0054] (1) Component 1, wherein Component 1 is prepared by the following steps: 55% of allantoin, 35% of glycerol, 8% of hyaluronic acid, and 2% of hydroxypropyl methylcellulose are mixed in a mass ratio, and stirred at 100 r / min for 1 hour to obtain; and

[0055] (2) Component 2, wherein Component 2 is prepared by the following steps: dissolving 1 g of a polypeptide having an amino acid sequence as shown in SEQ ID NO: 2 in 500 mL of an aqueous solution of 3 g of a polypeptide having an amino acid sequence as shown in SEQ ID NO: 1, adding 40 mg of poly-L-arginine, and stirring at room temperature for 5 h; ultrafiltration of the reaction solution using an NMWL 30 kDa ultrafiltration membrane, adjusting the pH value to 5.5 using dilute hydrochloric acid, adding 20 mg of zinc lactate, and freeze-drying to obtain the component.

[0056] The present invention provides a composition, characterized in that the composition comprises:

[0057] (1) Component 1, wherein Component 1 is prepared by the following steps: 55% of allantoin, 35% of glycerol, 8% of hyaluronic acid, and 2% of hydroxypropyl methylcellulose are mixed in a mass ratio, and stirred at 100 r / min for 1 hour to obtain; and

[0058] (2) Component 2, wherein Component 2 is prepared by the following steps: dissolving 1 g of a polypeptide having an amino acid sequence as shown in SEQ ID NO: 2 in 500 mL of an aqueous solution of 3 g of a polypeptide having an amino acid sequence as shown in SEQ ID NO: 1, adding 40 mg of poly-L-arginine, and stirring at room temperature for 5 h; ultrafiltration of the reaction solution using an NMWL 30 kDa ultrafiltration membrane, adjusting the pH value to 5.5 using dilute hydrochloric acid, adding 20 mg of zinc lactate, and freeze-drying to obtain the component.

[0059] In some embodiments of the present invention, the composition increases the permeability of the active ingredient of the drug.

[0060] In some embodiments of the present invention, the composition increases the penetration rate of active ingredients of cosmetics.

[0061] In some embodiments of the present invention, the method of using the composition is: apply component 1 to the surface of the skin, then wipe off component 1 and apply component 2. In some embodiments of the present invention, the method of using the composition is: apply component 1 to the surface of the skin, then wipe off component 1 and apply component 2 multiple times. In some embodiments of the present invention, the method of using the composition is: apply component 1 to the surface of the skin, wipe off component 1 after 10-15 minutes and start applying component 2. In some embodiments of the present invention, the method of using the composition is: apply component 1 to the surface of the skin, wipe off component 1 after 15 minutes and start applying component 2. In some embodiments of the present invention, the method of using the composition is: apply component 1 to the surface of the skin, wipe off component 1 after 10-15 minutes and start applying component 2 multiple times. Further, the multiple times are 2 times, 3 times or more times. Further, the multiple times are 2 times.

[0062] In some embodiments of the present invention, component 1 is applied to the skin surface, and component 2 is then applied every 4-5 hours.

[0063] In a second aspect of the present invention, there is provided use of the composition according to the first aspect of the present invention in improving the permeability of active ingredients in medicines or cosmetics.

[0064] In some embodiments of the present invention, there is provided use of the composition according to the first aspect of the present invention for improving the permeability of an active ingredient of a drug.

[0065] In some embodiments of the present invention, there is provided a use of the composition according to the first aspect of the present invention for improving the permeability of active ingredients in cosmetics.

[0066] In some embodiments of the present invention, the drug or cosmetic is applied to the skin surface.

[0067] In a third aspect of the present invention, a method for increasing the permeability of active ingredients of drugs or cosmetics is provided, characterized in that the method comprises applying the composition as described in the first aspect of the present invention to the skin surface.

[0068] The present invention provides a method for improving the permeability of active pharmaceutical ingredients, characterized in that the method comprises applying the composition as described in the first aspect of the present invention to the skin surface.

[0069] The present invention provides a method for improving the permeability of active ingredients in cosmetics, characterized in that the method comprises applying the composition as described in the first aspect of the present invention to the skin surface.

[0070] In some embodiments of the present invention, component 1 is applied to the surface of the skin, then component 1 is wiped off and component 2 is applied. In some embodiments of the present invention, component 1 is applied to the surface of the skin, then component 1 is wiped off and component 2 is applied multiple times. In some embodiments of the present invention, component 1 is applied to the surface of the skin, component 1 is wiped off after 10-15 minutes and component 2 is applied. In some embodiments of the present invention, component 1 is applied to the surface of the skin, component 1 is wiped off after 15 minutes and component 2 is applied. In some embodiments of the present invention, component 1 is applied to the surface of the skin, component 1 is wiped off after 10-15 minutes and component 2 is applied multiple times. Further, the multiple times are 2 times, 3 times or more times. Further, the multiple times are 2 times.

[0071] In some embodiments of the present invention, component 1 is applied to the skin surface, and component 2 is then applied every 4-5 hours.

[0072] In a fourth aspect of the present invention, a polypeptide is provided, wherein the amino acid sequence of the polypeptide is as shown in SEQ ID NO:1.

[0073] In the fifth aspect of the present invention, a nucleic acid is provided, characterized in that the nucleic acid encodes a polypeptide having an amino acid sequence as shown in SEQ ID NO:1.

[0074] The technical effect of the technical solution of the present invention: The composition of the present invention can greatly improve the skin permeability of active ingredients of drugs or cosmetics, achieving unexpected technical effects. The action time can be prolonged by slowly releasing the active ingredients, and the action time can also be prolonged by applying the component 2 multiple times. DETAILED DESCRIPTION

[0075] The permeability of the present invention may refer to the ratio of the active ingredient entering the skin to all active ingredients (in various forms, such as free active ingredients or bound active ingredients) applied to the skin surface. The ratio may be a mass ratio or a molar ratio.

[0076] The present invention is further described below by way of examples, but the present invention is not limited to the scope of the examples. The experimental methods in the following examples without specifying specific conditions are selected according to conventional methods and conditions, or according to the product specifications. Unless otherwise specified, the reagents and raw materials used in the present invention are all commercially available.

[0077] Example

[0078] Example 1. Preparation of polypeptide SCP-01

[0079] According to the method described in Angell, YM, et al. "Practical protocols for stepwise solid-phase synthesis of cysteine-containing peptides." The journal of peptideresearch 60.5 (2002): 292-299. Page 295, paragraph 2, the crude peptide was synthesized and purified by the Fmoc method. Then, the peptide SCP-01 (SEQ ID NO: 1) was obtained by purification by C18 HPLC preparative column (the mobile phase was acetonitrile containing 0.1% TFA and water containing 0.1% TFA). HPLC analysis showed a purity of >98.1%. The mass spectrum showed the target product, containing [M+3H] 3H+ 985.6 main ion peak.

[0080] Example 2: Preparation of Component 1 and Component 2

[0081] 55% of allantoin (Supelco, PHR1364), 35% of glycerol, 8% of hyaluronic acid (Sigma-Aldrich, Product Number 924474), and 2% of hydroxypropyl methylcellulose (HPMC) were mixed in a mass ratio to a total of 100 g. The mixture was stirred at 100 r / min for 1 hour to obtain a paste of component 1, which was recorded as component 1a.

[0082] 55% of allantoin (Supelco, PHR1364), 35% of glycerol, 8% of D-trehalose (Aladdin, product number D110019), and 2% of hydroxypropyl methylcellulose (HPMC) were mixed in a mass ratio to a total of 100 g. The mixture was stirred at 100 r / min for 1 hour to obtain a paste of component 1, which was recorded as component 1b.

[0083] The active ingredient is dissolved in an aqueous solution containing a thiol compound, polyarginine or polylysine is added, and the mixture is stirred. The reaction solution is dialyzed or ultrafiltered, the pH value is adjusted, zinc lactate is added, and the mixture is freeze-dried to obtain component 2.

[0084] Specifically, 1 g of active ingredient (polypeptide GMCCSR of patent CN105131086B, or polypeptide AHK-Cu [Special Peptide Biology, No. 411881]) was dissolved in 500 mL of aqueous solution of 3 g of polythiol compound (hydrolyzed keratin polypeptide [Discoverls], or hydrolyzed keratin [Puriactives, BioActive HK20 hydrolyzed keratin], or polypeptide SCP-01 [SEQ ID NO: 1], or dithiopropanol [Leyan, product No. 1083161]), 40 mg of poly-L-arginine (Sigma-Aldrich, P4663, mol wt 5000-15000) was added, and the mixture was stirred at room temperature for 5 h to form disulfide bonds between the active ingredient and the polythiol compound. Ultrafiltration membrane (Millipore, XL50with The reaction solution was ultrafiltered through a 30 kDa membrane, the pH value was adjusted to 5.5 with dilute hydrochloric acid, and 20 mg of zinc lactate was added. Freeze-dried. Components 2a-2h were obtained. The difference between the control components 2m and 2n and the above 2a-2h was that no polythiol compound was added, component 2m was prepared from the active ingredient polypeptide GMCCSR, and component 2n was prepared from the active ingredient polypeptide AHK-Cu. As shown in Table 1.

[0085] Table 1. Components 2 (2a-2p) prepared under different reaction conditions.

[0086]

[0087]

[0088] Example 3: Skin penetration test (diffusion cell method)

[0089] The test was conducted according to the in vitro transdermal experiment method reported in the literature (Ruan Wenyi, Qin Mengyao, Wan Tao, Long Yueming, Pan Jingtong, Wu Chuanbin, Wang Yinping, Xu Yuehong. Study on the effect of asarum volatile oil on the percutaneous penetration of sinapinic acid and its permeation-enhancing mechanism [J]. Chinese Herbal Medicine, 2017, 48(11): 2197-2201). 0.3 g of component 1a or component 1b prepared by the method described in Example 2 was first evenly applied to the surface of the stratum corneum of the skin, and component 1a or component 1b was gently wiped off after 10 minutes; and the aqueous dispersion of component 2 (2a-2h, 2m) prepared by the method described in Example 2 (0.3 g of component 2, evenly dispersed in 0.1 mL of water) was immediately evenly applied to the surface of the stratum corneum of the skin, and sealed with a sealing film. After 4 hours, the sealing film was removed, and the same aqueous dispersion of component 2 (2a-2h, 2m) (0.3g component 2, evenly dispersed in 0.1mL water) was applied to the surface of the stratum corneum of the skin again, and the surface was sealed with a sealing film. The control groups NC1 and NC2 were set up; components 1a and 1b were not applied, and the NC1 group only applied the aqueous dispersion of component 2m twice (0.3g component 2m, evenly dispersed in 0.1mL water), and the NC2 group only applied the aqueous dispersion of component 2n twice (0.3g component 2n, evenly dispersed in 0.1mL water). After 5 hours, the concentration of active ingredients in the receiving solution of each group was determined by C18 HPLC (the mobile phase was acetonitrile containing 0.1% TFA and water containing 0.1% TFA). Calculate the multiple of the active ingredient concentration in the receiving solution compared to the corresponding NC group (NC1 or NC2) (the multiple of the active ingredient being the peptide GMCCSR compared to the NC1 group, the multiple of the active ingredient being the peptide AHK-Cu compared to the NC2 group). Repeat the test 5 times and take the average value of the results. As shown in Table 2.

[0090] The concentration of active ingredients in the receiving solution of Scheme 1 and Scheme 5 was much higher than that of other groups, with statistically significant differences (P<0.01), indicating strong skin penetration ability. The concentration of active ingredients in the receiving solution of Scheme 2 and Scheme 3 was significantly higher than that of Scheme 4 (P<0.01), indicating that the effect of hydrolyzed keratin polypeptide was better than that of dithiopropanol. And the solution of component 1a was better than the corresponding solution of component 1b, indicating that the effect of the combination of allantoin-glycerol-hyaluronic acid and component 2 was better than the combination of allantoin-glycerol-D-trehalose and component 2.

[0091] Table 2. Skin penetration test results

[0092]

[0093]

[0094] Example 4: Live Animal Skin Permeability Test

[0095] Adult C57 mice were randomly divided into groups, with 10 mice in each group. A 10 mm × 5 mm area was taken from the upper left back of the mouse and shaved. 0.35 g of component 1a prepared as described in Example 2 was evenly applied to the surface of the skin, and component 1a was gently wiped off after 15 minutes; immediately, the aqueous dispersion of component 2 (2a, 2b, 2m) prepared as described in Example 2 (0.3 g component 2, evenly dispersed in 0.1 mL water) was evenly applied to the surface of the skin, and 4 hours later, the same aqueous dispersion of component 2 (0.3 g component 2, evenly dispersed in 0.1 mL water) was evenly applied to the surface of the skin for the second time. After another 4 hours, the same aqueous dispersion of component 2 (0.3 g component 2, evenly dispersed in 0.1 mL water) was evenly applied to the surface of the skin for the third time. After 5 hours, the mouse was killed, the applied area was gently wiped clean with a dry cloth, and the skin sample was carefully cut off from the applied area using a scalpel. First, the epidermis was peeled off to obtain an epidermis sample, and then the dermis was cut to obtain a dermis sample. The sample was minced and placed in a centrifuge tube containing PBS buffer (pH 7.2). The sample was evenly suspended using an ultrasonic processor and shaken for 30 minutes to extract the polypeptide GMCCSR. Gauze was filtered and then ultrafiltered using an ultrafiltration membrane (Millipore PLCC07610 5K). The filtrate was measured for the concentration of the active ingredient polypeptide GMCCSR using C18 HPLC (the mobile phase was acetonitrile containing 0.1% TFA and water containing 0.1% TFA).

[0096] The peptide GMCCSR was detected in both the epidermis and dermis, but the concentration difference was very large; the absolute value of the peptide GMCCSR concentration in the dermis samples of group 3 was much lower than that of the epidermis samples (about 1 / 34 of the former). The multiples of the peptide GMCCSR concentration of the epidermal samples of each group relative to the peptide GMCCSR concentration of the epidermal samples of group 3, and the multiples of the peptide GMCCSR concentration of the dermal samples of each group relative to the peptide GMCCSR concentration of the dermal samples of group 3 were calculated, and the results were taken as the average of each animal in the group. As shown in Table 3. The peptide GMCCSR concentrations in the epidermis and dermis samples of group 1 were much higher than those in groups 2 and 3, with statistically significant differences (P<0.01). The peptide GMCCSR concentrations in the epidermis and dermis samples of group 2 were much higher than those in group 3 (P<0.01).

[0097] Table 3. Animal experimental test results

[0098]

[0099] Example 5: Safety Test

[0100] In accordance with the "Technical Specifications for Safety of Cosmetics 2022 Edition (Draft for Comments)", acute dermal toxicity tests of components 1a, 2a, and 2b prepared as described in the preceding examples were carried out in adult rats, and the results showed that they were all slightly toxic; skin irritation / corrosion tests of components 1a, 2a, and 2b prepared as described in the preceding examples were carried out in rabbits, and the results showed that they were all non-irritating.

[0101] Although the embodiments of the present disclosure have been described above, it is understood that the above embodiments are exemplary and do not constitute limitations to the present invention. Those skilled in the art may change, modify and vary the above embodiments within the scope of the present invention.

Claims

1. A composition for improving the skin permeability of active ingredients of drugs or cosmetics, characterized in that: The composition comprises: (1) Component 1, wherein Component 1 is prepared by the following steps: 55% of allantoin, 35% of glycerol, 8% of hyaluronic acid, and 2% of hydroxypropyl methylcellulose are mixed in a mass ratio, and stirred at 100 r / min for 1 hour to obtain; and (2) Component 2, wherein Component 2 is prepared by the following steps: dissolving 1 g of a polypeptide having an amino acid sequence as shown in SEQ ID NO: 2 in 500 mL of an aqueous solution containing 3 g of a polypeptide having an amino acid sequence as shown in SEQ ID NO: 1, adding 40 mg of poly-L-arginine, and stirring the mixture at room temperature for 5 h; ultrafiltration of the reaction mixture using an NMWL 30 kDa ultrafiltration membrane, adjusting the pH value to 5.5 using dilute hydrochloric acid, adding 20 mg of zinc lactate, and freeze-drying the mixture to obtain the product.

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