Supramolecular collagen-sodium hyaluronate eutectic, supramolecular collagen-sodium hyaluronate composition, and preparation method and application of supramolecular collagen-sodium hyaluronate eutectic

Abstract

The invention relates to the technical field of supramolecules, and discloses a supramolecular collagen-sodium hyaluronate eutectic crystal, a supramolecular collagen-sodium hyaluronate composition, and a preparation method and application thereof. The preparation method of the supramolecular collagen-sodium hyaluronate eutectic comprises the following steps: respectively dissolving collagen and sodium hyaluronate in a solvent to obtain a collagen solution and a sodium hyaluronate solution, mixing the collagen solution and the sodium hyaluronate solution, heating the mixed solution, and carrying out eutectic reaction to obtain a supramolecular collagen-sodium hyaluronate solution; and concentrating, recrystallizing, filtering and drying to obtain the supramolecular collagen-sodium hyaluronate eutectic crystal. The supramolecular collagen-sodium hyaluronate eutectic crystal provided by the invention completely retains the main molecular skeleton of collagen, and also retains the biological activity and efficacy of sodium hyaluronate at the same time. According to the supramolecular collagen-sodium hyaluronate eutectic crystal, the solubility, skin permeability and bioavailability of collagen are improved, and the application effect of the supramolecular collagen-sodium hyaluronate eutectic crystal is remarkably enhanced.

Description

Technical Field

[0001] The present invention relates to the field of supramolecular technology, and particularly to a supramolecular collagen-sodium hyaluronate eutectic, a composition, a preparation method and applications thereof. Background Art

[0002] Collagen has the effects of moisturizing the skin, delaying aging, and anti-wrinkle beauty, and is very important for human health and beauty. Sodium hyaluronate, also known as hyaluronic acid, is a linear acidic mucopolysaccharide that plays important physiological functions such as water retention, lubrication, and promotion of cell repair in the intercellular matrix of human cells, the vitreous humor of the eye, and synovial fluid of joints.

[0003] Although collagen has been widely used in the fields of cosmetics, medical beauty, etc. due to its unique biocompatibility and functional characteristics of promoting tissue repair, its inherent macromolecular spatial conformation and physicochemical properties have led to its permeability becoming a technical problem that has not been overcome.

[0004] Collagen and sodium hyaluronate, as common raw materials for cosmetics and medical beauty, can generally be used in combination, but they are only physically mixed or compounded, and there is no intermolecular force between them, so the transdermal effect of collagen cannot be significantly improved.

[0005] Therefore, the existing technologies for improving the permeability of collagen still need to be improved and developed. Summary of the Invention

[0006] In view of the above deficiencies of the prior art, the purpose of the present invention is to provide a supramolecular collagen-sodium hyaluronate eutectic, a composition, a preparation method and applications thereof, aiming to solve the problem that the transdermal effect and bioavailability of collagen and sodium hyaluronate cannot be improved by physical mixing or compounding in the prior art.

[0007] The technical solution of the present invention is as follows:

[0008] In the first aspect of the present invention, a preparation method of a supramolecular collagen-sodium hyaluronate eutectic is provided, including the steps of:

[0009] Dissolving collagen and sodium hyaluronate in a solvent respectively to obtain a collagen solution and a sodium hyaluronate solution;

[0010] Mixing the collagen solution and the sodium hyaluronate solution, heating the mixed solution for eutectic reaction to obtain a supramolecular collagen-sodium hyaluronate solution;

[0011] Concentrating, recrystallizing, filtering and drying the supramolecular collagen-sodium hyaluronate solution to obtain a supramolecular eutectic constructed by collagen and sodium hyaluronate, that is, the supramolecular collagen-sodium hyaluronate eutectic.

[0012] Optionally, the molar ratio of the collagen to the sodium hyaluronate is 1:40 to 40:1.

[0013] Preferably, the molar ratio of the collagen to the sodium hyaluronate is 1:1, 1:2, 1:3, 1:4, 1:5, 1:6, 1:7, 1:8, 1:9, 1:10, 1:20, 1:30, 1:40, 2:1, 3:1, 4:1, 5:1, 6:1, 7:1, 8:1, 9:1, 10:1, 20:1, 30:1 or 40:1.

[0014] Preferably, the mixed solution is heated under an inert atmosphere to carry out a eutectic reaction.

[0015] Optionally, the temperature of the eutectic reaction is 30°C to 100°C.

[0016] Optionally, the time of the eutectic reaction is 4 h to 48 h.

[0017] Optionally, the recrystallization method is vacuum distillation.

[0018] Optionally, the drying method is freeze-drying. Preferably, the temperature of the freeze-drying is -100°C to -30°C, and the time of the freeze-drying is 4 h to 48 h.

[0019] In the second aspect of the present invention, there is provided a supramolecular collagen-sodium hyaluronate eutectic, which is prepared by the preparation method of the supramolecular collagen-sodium hyaluronate eutectic, and / or the supramolecular collagen-sodium hyaluronate eutectic is obtained by combining collagen and sodium hyaluronate through non-covalent bond forces (hydrogen bonds and / or intermolecular forces).

[0020] Optionally, the particle size of the supramolecular collagen-sodium hyaluronate eutectic is 1 nm to 500 nm.

[0021] In the third aspect of the present invention, there is provided a composition, and the composition includes the supramolecular collagen-sodium hyaluronate eutectic as an active ingredient.

[0022] In the fourth aspect of the present invention, there is provided an application of the composition in the preparation of cosmetics, foods, drugs or medical device products.

[0023] Beneficial effects: After the co-crystallization reaction of collagen and sodium hyaluronate in solution, the present invention generates a supramolecular collagen-sodium hyaluronate co-crystalline solution, and after concentration, recrystallization and drying, a supramolecular collagen-sodium hyaluronate co-crystal with relatively high purity is obtained. The supramolecular collagen-sodium hyaluronate co-crystal not only completely retains the main molecular skeleton and functional groups of collagen, but also retains the characteristics of sodium hyaluronate. Therefore, the supramolecular collagen-sodium hyaluronate co-crystal simultaneously has the biological activities and effects of the two active substances, and its nano-scale supramolecular structure can bypass the screening of the skin cutin layer and directly penetrate into the deep layer of the skin, significantly enhancing its biocompatibility, mildness, solubility, bioavailability and skin permeability. Description of the drawings

[0024] Figure 1 It is a schematic flow chart of the preparation method of the supramolecular collagen-sodium hyaluronate co-crystal provided by the present invention.

[0025] Figure 2 It is the 1H NMR spectrum of the supramolecular collagen-sodium hyaluronate co-crystal (10:1) prepared in Example 1 of the present invention.

[0026] Figure 3 It is the 2D 1H NMR spectrum of the supramolecular collagen-sodium hyaluronate co-crystal (10:1) prepared in Example 1 of the present invention.

[0027] Figure 4 It is the 1H NMR spectrum of the supramolecular collagen-sodium hyaluronate co-crystal (1:1) prepared in Example 2 of the present invention.

[0028] Figure 5 It is the 2D 1H NMR spectrum of the supramolecular collagen-sodium hyaluronate co-crystal (1:1) prepared in Example 2 of the present invention.

[0029] Figure 6 It is the 1H NMR spectrum of the supramolecular collagen-sodium hyaluronate co-crystal (40:1) prepared in Example 3 of the present invention.

[0030] Figure 7 It is the 2D 1H NMR spectrum of the supramolecular collagen-sodium hyaluronate co-crystal (40:1) prepared in Example 3 of the present invention.

[0031] Figure 8 It is the 1H NMR spectrum of the supramolecular collagen-sodium hyaluronate co-crystal (10:1, without nitrogen) prepared in Example 4 of the present invention.

[0032] Figure 9 It is the 2D 1H NMR spectrum of the supramolecular collagen-sodium hyaluronate co-crystal (10:1, without nitrogen) prepared in Example 4 of the present invention.

[0033] Figure 10 1H NMR spectrum of the supramolecular collagen-sodium hyaluronate eutectic (1:1, without nitrogen) prepared in Example 5 of the present invention.

[0034] Figure 11 2D 1H NMR spectrum of the supramolecular collagen-sodium hyaluronate eutectic (1:1, without nitrogen) prepared in Example 5 of the present invention.

[0035] Figure 12 1H NMR spectrum of the supramolecular collagen-sodium hyaluronate eutectic (40:1, without nitrogen) prepared in Example 6 of the present invention.

[0036] Figure 13 2D 1H NMR spectrum of the supramolecular collagen-sodium hyaluronate eutectic (40:1, without nitrogen) prepared in Example 6 of the present invention.

[0037] Figure 14 1H NMR spectrum of the supramolecular collagen-sodium hyaluronate eutectic (1:50) prepared in Example 7 of the present invention.

[0038] Figure 15 2D 1H NMR spectrum of the supramolecular collagen-sodium hyaluronate eutectic (1:50) prepared in Example 7 of the present invention.

[0039] Figure 16 1H NMR spectrum of the supramolecular collagen-sodium hyaluronate eutectic (1:50, without nitrogen) prepared in Example 8 of the present invention.

[0040] Figure 17 2D 1H NMR spectrum of the supramolecular collagen-sodium hyaluronate eutectic (1:50, without nitrogen) prepared in Example 8 of the present invention.

[0041] Figure 18Transmission electron micrographs of the supramolecular collagen-sodium hyaluronate eutectics prepared in the embodiments of the present invention. Among them, (a) is the supramolecular collagen-sodium hyaluronate eutectic (10:1) prepared in Example 1, (b) is the supramolecular collagen-sodium hyaluronate eutectic (1:1) prepared in Example 2, (c) is the supramolecular collagen-sodium hyaluronate eutectic (40:1) prepared in Example 3, (d) is the supramolecular collagen-sodium hyaluronate eutectic (10:1, without nitrogen) prepared in Example 4, (e) is the supramolecular collagen-sodium hyaluronate eutectic (1:1, without nitrogen) prepared in Example 5, (f) is the supramolecular collagen-sodium hyaluronate eutectic (40:1, without nitrogen) prepared in Example 6, (g) is the supramolecular collagen-sodium hyaluronate eutectic (1:50) prepared in Example 7, and (h) is the supramolecular collagen-sodium hyaluronate eutectic (1:50, without nitrogen) prepared in Example 8.

[0042] Figure 19 Transdermal efficiency graph of the supramolecular collagen-sodium hyaluronate eutectics prepared in the embodiments of the present invention.

[0043] Figure 20 Result graph of the effect of the supramolecular collagen-sodium hyaluronate eutectic (10:1) prepared in Example 1 of the present invention on the chondroitin sulfate content.

[0044] Figure 21 Result graph of the effect of the supramolecular collagen-sodium hyaluronate eutectic (10:1) prepared in Example 1 of the present invention on the Collagen-I content.

[0045] Figure 22 Result graph of the effect of the supramolecular collagen-sodium hyaluronate eutectic (10:1) prepared in Example 1 of the present invention on the Collagen-III content.

[0046] Figure 23 Result graph of the effect of the supramolecular collagen-sodium hyaluronate eutectic (10:1) prepared in Example 1 of the present invention on the thickness of the epidermal living cell layer. Detailed Description of the Invention

[0047] The present invention provides a supramolecular collagen-sodium hyaluronate eutectic, a composition, a preparation method and applications thereof. To make the objectives, technical solutions and effects of the present invention clearer and more definite, the present invention is further described in detail below. It should be understood that the specific embodiments described herein are only used to explain the present invention and are not used to limit the present invention.

[0048] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the technical field to which this invention belongs. The terms used in the specification of this invention are for the purpose of describing specific embodiments only and are not intended to limit the invention.

[0049] An embodiment of the present invention provides a method for preparing a supramolecular collagen-sodium hyaluronate eutectic, as Figure 1 shown, comprising the steps of:

[0050] S1. Dissolve collagen and sodium hyaluronate in a solvent respectively to obtain a collagen solution and a sodium hyaluronate solution;

[0051] S2. Mix the collagen solution and the sodium hyaluronate solution, heat the mixed solution for eutectic reaction to obtain a supramolecular collagen-sodium hyaluronate solution;

[0052] S3. Concentrate, recrystallize, filter, and dry the supramolecular collagen-sodium hyaluronate solution to obtain a supramolecular eutectic constructed by collagen and sodium hyaluronate, that is, the supramolecular collagen-sodium hyaluronate eutectic.

[0053] The method for preparing the supramolecular collagen-sodium hyaluronate eutectic provided by the present invention uses easily available solvents such as ethanol or water as the reaction medium. The eutectic reaction conditions are easy to achieve, the synthesis steps are simple, and the post-treatment is convenient. The obtained supramolecular collagen product has high purity and high yield. The supramolecular collagen-sodium hyaluronate eutectic has excellent biocompatibility, mildness, solubility, high bioavailability and skin permeability. Moreover, the supramolecular collagen-sodium hyaluronate eutectic not only completely retains the main molecular skeleton of collagen, but also retains the biological activity and efficacy of the eutectic ligand, greatly improving the skin permeability and bioavailability of collagen and enhancing its application effect. In addition, the supramolecular collagen-sodium hyaluronate eutectic generally refers to an aggregate in which collagen molecules and sodium hyaluronate molecules are combined together by intermolecular interactions, forming a complex and organized aggregate and maintaining a certain integrity, so that it has a clear microstructure and macroscopic properties. The supramolecular collagen-sodium hyaluronate eutectic provided by the present invention can directly penetrate into the deep layer of the skin by bypassing the screening of the stratum corneum due to its unique nano-scale supramolecular structure, and can maintain the stability of carrying active substances, so it can maintain its fresh activity for a long time.

[0054] In step S1, in some embodiments, the molar ratio of the collagen to the sodium hyaluronate is 1:40 to 40:1.

[0055] In some preferred embodiments, the molar ratio of the collagen to the sodium hyaluronate is 1:1, 1:2, 1:3, 1:4, 1:5, 1:6, 1:7, 1:8, 1:9, 1:10, 1:20, 1:30, 1:40, 2:1, 3:1, 4:1, 5:1, 6:1, 7:1, 8:1, 9:1, 10:1, 20:1, 30:1 or 40:1.

[0056] In some preferred embodiments, the molar ratio of the collagen to the sodium hyaluronate is 10:1.

[0057] In some embodiments, the solvent is selected from one or two of ethanol, water, methanol, isopropanol, acetone, propylene glycol, butylene glycol and pentylene glycol.

[0058] In step S2, in some embodiments, the mixed solution is heated under an inert atmosphere (such as a nitrogen atmosphere, etc.) for a eutectic reaction. Under an inert atmosphere, the formed supramolecular collagen-sodium hyaluronate eutectic can be made more stable.

[0059] In some embodiments, the temperature of the eutectic reaction is 30°C to 100°C, for example, it can be 30°C, 40°C, 50°C, 60°C, 70°C, 80°C, 90°C or 100°C, etc.

[0060] In some embodiments, the time of the eutectic reaction is 4h to 48h, for example, it can be 4h, 5h, 8h, 10h, 15h, 20h, 25h, 30h, 35h, 40h, 45h or 48h, etc.

[0061] In step S3, in some embodiments, the recrystallization method is vacuum distillation.

[0062] In some embodiments, the drying method is freeze-drying. Preferably, the temperature of the freeze-drying is -100°C to -30°C (for example, it can be -100°C, -90°C, -80°C, -70°C, -60°C, -50°C, -40°C or -30°C, etc.), and the time of the freeze-drying is 4h to 48h (for example, it can be 4h, 5h, 8h, 10h, 15h, 20h, 25h, 30h, 35h, 40h, 45h or 48h, etc.), so as to completely remove the remaining water or solvents such as ethanol in the product.

[0063] The embodiment of the present invention provides a supramolecular collagen-sodium hyaluronate eutectic, which is prepared by the preparation method of the supramolecular collagen-sodium hyaluronate eutectic. The supramolecular collagen-sodium hyaluronate eutectic is obtained by combining collagen and sodium hyaluronate through non-covalent bond forces (hydrogen bonds and / or intermolecular forces).

[0064] Collagen has the effects of moisturizing the skin, delaying aging, and anti-wrinkle beauty, which is very important for human health and beauty. However, as a macromolecule, the permeability of collagen is a technical problem that has not been overcome. For physically mixed or compounded collagen and sodium hyaluronate, intermolecular forces cannot be formed, so the transdermal effect cannot be significantly improved. The supramolecular collagen-sodium hyaluronate eutectic prepared by the present invention has better biocompatibility, mildness, solubility, high bioavailability and skin permeability. Moreover, the supramolecular collagen-sodium hyaluronate eutectic not only completely retains the main molecular skeleton of collagen, but also retains the biological activity and efficacy of sodium hyaluronate. Collagen and sodium hyaluronate are combined through hydrogen bonds or intermolecular forces to obtain the supramolecular collagen-sodium hyaluronate eutectic, which can not only solve the problem of poor permeability of collagen, but also improve its stability and bioavailability.

[0065] In some embodiments, the particle size of the supramolecular collagen-sodium hyaluronate eutectic is 1 nm to 500 nm; preferably, the particle size of the supramolecular collagen-sodium hyaluronate eutectic is 18 nm to 44 nm.

[0066] The embodiment of the present invention provides a composition, and the active ingredient of the composition includes the supramolecular collagen-sodium hyaluronate eutectic described above.

[0067] In some specific embodiments, the composition further includes a cosmetically acceptable carrier or excipient, such as water, glycerin, petrolatum, alcohols, and oils, etc.

[0068] The embodiment of the present invention also provides an application of the composition described above in the preparation of cosmetics (such as skin care products, etc.), foods (such as health products, etc.), drugs or medical device products (such as tissue repair products, etc.).

[0069] The supramolecular collagen-sodium hyaluronate eutectic prepared in the embodiment of the present invention can be used as a formula raw material for cosmetics, medicine and life sciences, or as a medicinal ingredient or a raw material for a drug delivery carrier for transdermal drug delivery, etc. The supramolecular collagen-sodium hyaluronate eutectic not only completely retains the main molecular skeleton and functional groups of collagen, but also retains the characteristics of sodium hyaluronate. Therefore, it has the biological activities and effects of two active substances at the same time. For the transdermal absorption degree of collagen and the effects of transdermal drug delivery, etc., the supramolecular collagen-sodium hyaluronate eutectic provided by the present invention has a significant enhancing effect.

[0070] The following is a detailed description through specific examples.

[0071] Example 1

[0072] This example provides a method for preparing a supramolecular collagen-sodium hyaluronate eutectic (10:1), which includes the following steps:

[0073] Under a nitrogen atmosphere, dissolve 10 mol of collagen in 10 mL of water in a reactor to obtain a collagen solution; dissolve 1 mol of sodium hyaluronate in 10 mL of water, and dropwise add the obtained sodium hyaluronate solution into the reactor containing the collagen solution, heat to 40 °C, and carry out eutectic reaction for 24 hours.

[0074] After the reaction is completed, concentrate the supramolecular collagen-sodium hyaluronate solution to 1 / 10 of the reaction solution under vacuum conditions, carry out vacuum distillation crystallization, and separate through filtration and washing to obtain supramolecular collagen. Dry it in a vacuum drying oven for 48 hours to obtain a supramolecular collagen-sodium hyaluronate eutectic (10:1) with a purity exceeding 99%, and the yield is 95.37%.

[0075] Example 2

[0076] This example provides a method for preparing a supramolecular collagen-sodium hyaluronate eutectic (1:1), which includes the following steps:

[0077] Under a nitrogen atmosphere, dissolve 1 mol of collagen in 10 mL of water in a reactor to obtain a collagen solution; dissolve 1 mol of sodium hyaluronate in 10 mL of water, and dropwise add the obtained sodium hyaluronate solution into the reactor containing the collagen solution, heat to 40 °C, and carry out eutectic reaction for 24 hours.

[0078] After the reaction is completed, concentrate the supramolecular collagen-sodium hyaluronate solution to 1 / 10 of the reaction solution under vacuum conditions, carry out vacuum distillation crystallization, and separate through filtration and washing to obtain supramolecular collagen. Dry it in a vacuum drying oven for 48 hours to obtain a supramolecular collagen-sodium hyaluronate eutectic (1:1) with a purity exceeding 99%, and the yield is 93.27%.

[0079] Example 3

[0080] This example provides a method for preparing a supramolecular collagen-sodium hyaluronate eutectic (40:1), which includes the following steps:

[0081] Under a nitrogen atmosphere, dissolve 40 mol of collagen in 10 mL of water in a reactor to obtain a collagen solution; dissolve 1 mol of sodium hyaluronate in 10 mL of water, and dropwise add the obtained sodium hyaluronate solution into the reactor containing the collagen solution, heat to 40 °C, and carry out eutectic reaction for 24 hours.

[0082] After the reaction, the supramolecular collagen-sodium hyaluronate solution was concentrated to 1 / 10 of the reaction solution under vacuum conditions, crystallized by vacuum distillation under reduced pressure, and the supramolecular collagen was separated by filtration and washing. The supramolecular collagen-sodium hyaluronate eutectic (40:1) with a purity exceeding 99% was obtained after drying in a vacuum drying oven for 48 hours, and the yield was 93.46%.

[0083] Example 4

[0084] This example provides a method for preparing a supramolecular collagen-sodium hyaluronate eutectic (10:1, without nitrogen), which includes the following steps:

[0085] In an air atmosphere (i.e., not in a nitrogen atmosphere, such as in an indoor environment), 10 mol of collagen was dissolved in 10 mL of water in a reactor to obtain a collagen solution; 1 mol of sodium hyaluronate was dissolved in 10 mL of water, and the obtained sodium hyaluronate solution was added dropwise to the reactor containing the collagen solution, heated to 40 °C, and the eutectic reaction was carried out for 24 hours.

[0086] After the reaction, the supramolecular collagen-sodium hyaluronate solution was concentrated to 1 / 10 of the reaction solution under vacuum conditions, crystallized by vacuum distillation under reduced pressure, and the supramolecular collagen was separated by filtration and washing. The supramolecular collagen-sodium hyaluronate eutectic (10:1, without nitrogen) with a purity exceeding 99% was obtained after drying in a vacuum drying oven for 48 hours, and the yield was 95.25%.

[0087] Example 5

[0088] This example provides a method for preparing a supramolecular collagen-sodium hyaluronate eutectic (1:1, without nitrogen), which includes the following steps:

[0089] In an air atmosphere (i.e., not in a nitrogen atmosphere, such as in an indoor environment), 1 mol of collagen was dissolved in 10 mL of water in a reactor to obtain a collagen solution; 1 mol of sodium hyaluronate was dissolved in 10 mL of water, and the obtained sodium hyaluronate solution was added dropwise to the reactor containing the collagen solution, heated to 40 °C, and the eutectic reaction was carried out for 24 hours.

[0090] After the reaction, the supramolecular collagen-sodium hyaluronate solution was concentrated to 1 / 10 of the reaction solution under vacuum conditions, crystallized by vacuum distillation under reduced pressure, and the supramolecular collagen was separated by filtration and washing. The supramolecular collagen-sodium hyaluronate eutectic (1:1, without nitrogen) with a purity exceeding 99% was obtained after drying in a vacuum drying oven for 48 hours, and the yield was 93.27%.

[0091] Example 6

[0092] This example provides a preparation method of supramolecular collagen-sodium hyaluronate eutectic (40:1, without nitrogen), including the following steps:

[0093] Under an air atmosphere (i.e., not under a nitrogen atmosphere, such as in an indoor environment), dissolve 40 mol of collagen in 10 mL of water in a reactor to obtain a collagen solution; dissolve 1 mol of sodium hyaluronate in 10 mL of water, and dropwise add the obtained sodium hyaluronate solution into the reactor containing the collagen solution, heat to 40 °C, and carry out eutectic reaction for 24 hours.

[0094] After the reaction is completed, concentrate the supramolecular collagen-sodium hyaluronate solution to 1 / 10 of the reaction solution under vacuum conditions, carry out vacuum distillation crystallization, and separate through filtration and washing to obtain supramolecular collagen. Dry it in a vacuum drying oven for 48 hours to obtain supramolecular collagen-sodium hyaluronate eutectic (40:1, without nitrogen) with a purity exceeding 99%, and the yield is 93.46%.

[0095] Example 7

[0096] This example provides a preparation method of supramolecular collagen-sodium hyaluronate eutectic (1:50), including the following steps:

[0097] Under a nitrogen atmosphere, dissolve 1 mol of collagen in 10 mL of water in a reactor to obtain a collagen solution; dissolve 50 mol of sodium hyaluronate in 10 mL of water, and dropwise add the obtained sodium hyaluronate solution into the reactor containing the above collagen solution, heat to 40 °C, and carry out eutectic reaction for 24 hours.

[0098] After the reaction is completed, concentrate the supramolecular collagen-sodium hyaluronate solution to 1 / 10 of the reaction solution under vacuum conditions, carry out vacuum distillation crystallization, and separate through filtration and washing to obtain supramolecular collagen. Dry it in a vacuum drying oven for 48 hours to obtain supramolecular collagen-sodium hyaluronate eutectic (1:50), and the yield is 89.37%.

[0099] Example 8

[0100] This example provides a preparation method of supramolecular collagen-sodium hyaluronate eutectic (1:50, without nitrogen), including the following steps:

[0101] Under an air atmosphere (i.e., not under a nitrogen atmosphere, such as in an indoor environment), dissolve 1 mol of collagen in 10 mL of water in a reactor to obtain a collagen solution; dissolve 50 mol of sodium hyaluronate in 10 mL of water, and dropwise add the obtained sodium hyaluronate solution into the reactor containing the above collagen solution, heat to 40 °C, and carry out eutectic reaction for 24 hours.

[0102] After the reaction, the supramolecular collagen-sodium hyaluronate solution was concentrated to 1 / 10 of the reaction solution under vacuum conditions, distilled and crystallized under reduced pressure, and the supramolecular collagen was obtained by filtration and washing. It was dried in a vacuum drying oven for 48 hours to obtain the supramolecular collagen-sodium hyaluronate eutectic with a purity of (1:50, without nitrogen), and the yield was 89.37%. Test:

[0103] (1) The supramolecular collagen-sodium hyaluronate eutectic prepared in Examples 1-8 was subjected to nuclear magnetic resonance and melting point tests.

[0104] The 1H NMR spectrum of the supramolecular collagen-sodium hyaluronate eutectic (10:1) prepared in Example 1 is as Figure 2 shown, and the 1H NMR data are: 1 H NMR(400MHz,DMSO)δ8.52(s,2H),7.32(m,2H),7.19(s,2H),5.11(d,3H),4.52(t,1H),4.43(m,1H)4.35(m,6H),4.04(dd,3H),3.81(m,6H),3.61(m,4H),3.39(m,4H),3.23(m,2H),3.12(s,6H);2.91(s,4H),2.63(d,5H),2.24(t,2H),1.94(m,4H),1.59(s,4H),1.29(m,4H),1.12(d,4H),0.79(m,4H).

[0105] The 13C NMR data of the supramolecular collagen-sodium hyaluronate eutectic (10:1) prepared in Example 1 are: 13 CNMR(400MHz,DMSO)δ188.73(s),171.12(s),167.69(s),130.28(s),166.75(s),166.07(s),160.99(s),156.25(s),145.86(s),68.46(s),59.88(s),55.89(s),53.18(s),49.34(s),45.80(s),42.79(s),31.31(s),29.54(s),27.20(s),24.19(s),21.90(s),21.07(s),18.47(s),16.97(s),15.34(s),9.58(s).

[0106] The melting point of the supramolecular collagen-sodium hyaluronate eutectic (10:1) prepared in Example 1 was 210 °C.

[0107] The two-dimensional nuclear magnetic resonance hydrogen spectrum of the supramolecular collagen-sodium hyaluronate eutectic prepared in Example 1 is as follows Figure 3 As shown, it can be seen that there is a non-covalent interaction between collagen and sodium hyaluronate, indicating that a supramolecular structure is formed between collagen and sodium hyaluronate.

[0108] The nuclear magnetic resonance hydrogen spectrum of the supramolecular collagen-sodium hyaluronate eutectic (1:1) prepared in Example 2 is as follows Figure 4 As shown, its nuclear magnetic resonance hydrogen spectrum data are as follows: 1 H NMR(400MHz,DMSO)δ9.20(s,2H),8.36(m,2H),8.16(s,2H),7.61(d,3H),6.48(t,1H),5.74(m,1H)5.11(m,6H),4.83(dd,3H),4.41(m,6H),4.31(m,4H),3.86(m,4H),3.65(m,2H),3.44(s,6H);3.14(s,4H),3.03(d,5H),2.24(t,2H),1.94(m,4H),1.87(s,4H),1.55(m,4H),1.23(d,4H),0.80(m,4H).

[0109] The nuclear magnetic resonance carbon spectrum data of the supramolecular collagen-sodium hyaluronate eutectic (1:1) prepared in Example 2 are as follows: 13 CNMR(400MHz,DMSO)δ187.77(s),171.25(s),168.79(s),131.28(s),167.74(s),167.07(s),162.23(s),157.45(s),147.81(s),69.53(s),59.82(s),55.35(s),52.19(s),49.47(s),45.83(s),42.95(s),32.31(s),29.78(s),27.45(s),24.24(s),21.92(s),21.17(s),18.45(s),17.07(s),15.26(s),10.15(s).

[0110] The melting point of the supramolecular collagen-sodium hyaluronate eutectic (1:1) prepared in Example 2 is 214 °C.

[0111] The two-dimensional nuclear magnetic resonance hydrogen spectrum of the supramolecular collagen-sodium hyaluronate eutectic (1:1) prepared in Example 2 is as follows Figure 5 As shown, it can be seen that there is a weak non-covalent interaction between collagen and sodium hyaluronate, indicating that a supramolecular structure is formed between collagen and sodium hyaluronate, but it is unstable.

[0112] The 1H NMR spectrum of the supramolecular collagen-sodium hyaluronate eutectic (40:1) prepared in Example 3 is as follows Figure 6 shown, and its 1H NMR data are as follows: 1 1H NMR (400 MHz, DMSO) δ 8.72 (s, 2H), 8.37 (m, 2H), 8.17 (s, 2H), 7.91 (d, 3H), 7.62 (t, 1H), 7.30 (m, 1H), 6.84 (m, 6H), 4.49 (dd, 3H), 4.29 (m, 6H), 3.96 (m, 4H), 3.62 (m, 4H), 3.39 (m, 2H), 3.09 (s, 6H); 2.71 (s, 4H), 2.23 (d, 5H), 2.14 (t, 2H), 1.83 (m, 4H), 1.54 (s, 4H), 1.22 (m, 4H), 1.06 (d, 4H), 0.81 (m, 4H).

[0113] The 13C NMR data of the supramolecular collagen-sodium hyaluronate eutectic (40:1) prepared in Example 3 are as follows: 13 13C NMR (400 MHz, DMSO) δ 186.23 (s), 170.56 (s), 166.69 (s), 131.32 (s), 165.56 (s), 166.01 (s), 160.03 (s), 155.25 (s), 144.83 (s), 65.46 (s), 58.82 (s), 55.45 (s), 53.13 (s), 49.26 (s), 45.37 (s), 42.18 (s), 31.25 (s), 28.35 (s), 26.21 (s), 23.13 (s), 21.92 (s), 21.01 (s), 19.23 (s), 17.03 (s), 15.76 (s), 11.12 (s).

[0114] The melting point of the supramolecular collagen-sodium hyaluronate eutectic (40:1) prepared in Example 3 is 208 °C.

[0115] The 2D 1H NMR spectrum of the supramolecular collagen-sodium hyaluronate eutectic (40:1) prepared in Example 3 is as follows Figure 7 shown. It can be seen that there is a weak non-covalent interaction force between collagen and sodium hyaluronate, indicating that a supramolecular structure can be formed between collagen and sodium hyaluronate, but it is unstable.

[0116] The 1H NMR spectrum of the supramolecular collagen-sodium hyaluronate eutectic (10:1, without nitrogen) prepared in Example 4 is as follows Figure 8 shown, and its 1H NMR data are as follows: 11H NMR (400 MHz, DMSO) δ 8.63 (s, 2H), 8.50 (m, 2H), 8.24 (s, 2H), 7.99 (d, 3H), 7.64 (t, 1H), 7.33 (m, 1H) 6.87 (m, 6H), 4.46 (dd, 3H), 4.28 (m, 6H), 4.01 (m, 4H), 3.70 (m, 4H), 3.38 (m, 2H), 3.02 (s, 6H); 2.72 (s, 4H), 2.19 (d, 5H), 1.82 (t, 2H), 1.54 (m, 8H), 1.22 (s, 4H), 0.98 (d, 4H), 0.80 (m, 4H).

[0117] The carbon nuclear magnetic resonance data of the supramolecular collagen-sodium hyaluronate eutectic (10:1, without nitrogen) prepared in Example 4 are as follows: 13 13C NMR (400 MHz, DMSO) δ 188.73 (s), 172.19 (s), 168.63 (s), 131.36 (s), 167.83 (s), 167.25 (s), 161.4 (s), 155.27 (s), 146.83 (s), 69.36 (s), 60.88 (s), 55.45 (s), 53.27 (s), 49.38 (s), 45.81 (s), 42.35 (s), 31.57 (s), 29.24 (s), 28.10 (s), 25.13 (s), 20.92 (s), 19.07 (s), 18.21 (s), 16.23 (s), 15.04 (s), 9.88 (s).

[0118] The melting point of the supramolecular collagen-sodium hyaluronate eutectic (10:1, without nitrogen) prepared in Example 4 is 215 °C.

[0119] The two-dimensional nuclear magnetic resonance hydrogen spectrum of the supramolecular collagen-sodium hyaluronate eutectic (10:1, without nitrogen) prepared in Example 4 is as Figure 9 shown. It can be seen that there is a non-covalent interaction force between collagen and sodium hyaluronate, indicating that a supramolecular structure is formed between collagen and sodium hyaluronate.

[0120] The nuclear magnetic resonance hydrogen spectrum of the supramolecular collagen-sodium hyaluronate eutectic (1:1, without nitrogen) prepared in Example 5 is as Figure 10 shown, and its nuclear magnetic resonance hydrogen spectrum data are as follows: 11H NMR (400 MHz, DMSO) δ 9.13 (s, 2H), 8.45 (m, 2H), 8.63 (s, 2H), 7.68 (d, 3H), 6.54 (t, 1H), 5.73 (m, 1H), 5.12 (m, 6H), 4.86 (dd, 3H), 4.53 (m, 6H), 4.25 (m, 4H), 3.90 (m, 4H), 3.68 (m, 2H), 3.47 (s, 6H); 3.22 (s, 4H), 3.12 (d, 5H), 2.23 (t, 2H), 1.99 (m, 4H), 1.56 (s, 4H), 1.55 (m, 4H), 1.32 (d, 4H), 0.83 (m, 4H).

[0121] The carbon nuclear magnetic resonance data of the supramolecular collagen-sodium hyaluronate eutectic (1:1, without nitrogen) prepared in Example 5 are as follows: 13 13C NMR (400 MHz, DMSO) δ 185.73 (s), 170.32 (s), 169.73 (s), 161.28 (s), 160.74 (s), 160.07 (s), 159.23 (s), 157.45 (s), 147.88 (s), 69.59 (s), 59.45 (s), 55.25 (s), 52.20 (s), 49.47 (s), 45.81 (s), 42.23 (s), 32.45 (s), 29.78 (s), 27.45 (s), 24.24 (s), 21.92 (s), 20.24 (s), 18.46 (s), 17.25 (s), 15.45 (s), 10.63 (s).

[0122] The melting point of the supramolecular collagen-sodium hyaluronate eutectic (1:1, without nitrogen) prepared in Example 5 is 216 °C.

[0123] The two-dimensional nuclear magnetic resonance hydrogen spectrum of the supramolecular collagen-sodium hyaluronate eutectic (1:1, without nitrogen) prepared in Example 5 is as shown in Figure 11 It can be seen that there is a weak non-covalent interaction force between collagen and sodium hyaluronate, indicating that a supramolecular structure is formed between collagen and sodium hyaluronate, but it is unstable.

[0124] The nuclear magnetic resonance hydrogen spectrum of the supramolecular collagen-sodium hyaluronate eutectic (40:1, without nitrogen) prepared in Example 6 is as shown in Figure 12 It is shown, and its nuclear magnetic resonance hydrogen spectrum data are as follows: 11H NMR (400 MHz, DMSO) δ 8.71 (s, 2H), 8.43 (m, 2H), 8.15 (s, 2H), 7.87 (d, 3H), 7.56 (t, 1H), 7.33 (m, 1H) 6.76 (m, 6H), 4.54 (dd, 3H), 4.38 (m, 6H), 3.96 (m, 4H), 3.72 (m, 4H), 3.43 (m, 2H), 3.12 (s, 6H); 2.81 (s, 4H), 2.35 (d, 5H), 2.17 (t, 2H), 1.89 (m, 4H), 1.62 (s, 4H), 1.29 (m, 4H), 1.13 (d, 4H), 0.86 (m, 4H).

[0125] The carbon nuclear magnetic resonance spectrum data of the supramolecular collagen-sodium hyaluronate eutectic (40:1, without nitrogen) prepared in Example 6 are as follows: 13 13C NMR (400 MHz, DMSO) δ 185.23 (s), 171.56 (s), 166.45 (s), 161.32 (s), 160.56 (s), 160.01 (s), 159.03 (s), 155.25 (s), 145.83 (s), 67.46 (s), 59.85 (s), 55.48 (s), 54.35 (s), 50.34 (s), 45.39 (s), 42.24 (s), 31.35 (s), 28.46 (s), 26.38 (s), 22.25 (s), 21.83 (s), 21.12 (s), 19.17 (s), 17.35 (s), 16.06 (s), 11.28 (s).

[0126] The melting point of the supramolecular collagen-sodium hyaluronate eutectic (40:1) of the supramolecular collagen-sodium hyaluronate eutectic (40:1, without nitrogen) prepared in Example 6 is 204 °C.

[0127] The two-dimensional nuclear magnetic resonance hydrogen spectrum of the supramolecular collagen-sodium hyaluronate eutectic (40:1, without nitrogen) prepared in Example 6 is as follows Figure 13 shown. It can be seen that there is a weak non-covalent interaction force between collagen and sodium hyaluronate, indicating that a supramolecular structure can be formed between collagen and sodium hyaluronate, but it is unstable.

[0128] The nuclear magnetic resonance hydrogen spectrum of the supramolecular collagen-sodium hyaluronate eutectic (1:50) prepared in Example 7 is as follows Figure 14 shown, and its nuclear magnetic resonance hydrogen spectrum data are as follows: 11H NMR (400 MHz, DMSO) δ 9.10 (s, 2H), 8.68 (m, 2H), 8.45 (s, 2H), 8.36 (d, 3H), 6.52 (t, 1H), 5.84 (m, 1H), 5.25 (m, 6H), 5.02 (dd, 3H), 4.91 (m, 6H), 4.47 (m, 4H), 4.37 (m, 4H), 4.30 (m, 2H), 3.88 (s, 6H); 3.64 (s, 4H), 3.45 (d, 9H), 3.22 (t, 6H), 1.85 (m, 4H), 1.24 (s, 4H), 0.82 (m, 4H).

[0129] The carbon NMR data of the supramolecular collagen-sodium hyaluronate eutectic (1:50) prepared in Example 7 are as follows: 13 13C NMR (400 MHz, DMSO) δ 189.92 (s), 172.25 (s), 168.69 (s), 132.58 (s), 167.78 (s), 166.07 (s), 162.94 (s), 166.25 (s), 147.86 (s), 70.46 (s), 61.88 (s), 57.89 (s), 54.25 (s), 50.37 (s), 45.89 (s), 42.64 (s), 32.36 (s), 30.58 (s), 28.23 (s), 24.25 (s), 21.96 (s), 21.35 (s), 18.67 (s), 16.88 (s), 15.36 (s), 11.53 (s).

[0130] The melting point of the supramolecular collagen-sodium hyaluronate eutectic (1:50) prepared in Example 7 is 219 °C.

[0131] The two-dimensional 1H NMR spectrum of the supramolecular collagen-sodium hyaluronate eutectic (1:50) prepared in Example 7 is as Figure 15 shown. It can be seen that there is no non-covalent interaction between collagen and sodium hyaluronate, indicating that no supramolecular structure is formed between collagen and sodium hyaluronate.

[0132] The 1H NMR spectrum of the supramolecular collagen-sodium hyaluronate eutectic (1:50, without nitrogen) prepared in Example 8 is as Figure 16 shown, and its 1H NMR data are as follows: 11H NMR (400 MHz, DMSO) δ 9.08 (s, 2H), 8.62 (m, 2H), 8.33 (s, 2H), 8.32 (d, 3H), 6.14 (t, 1H), 5.56 (m, 1H), 5.28 (m, 6H), 5.12 (dd, 3H), 4.83 (m, 6H), 4.42 (m, 4H), 4.25 (m, 4H), 4.12 (m, 2H), 3.73 (s, 6H); 3.65 (s, 4H), 3.53 (d, 9H), 3.13 (t, 6H), 1.89 (m, 4H), 1.34 (s, 4H), 0.86 (m, 4H).

[0133] The carbon nuclear magnetic resonance data of the supramolecular collagen-sodium hyaluronate eutectic (1:50, without nitrogen) prepared in Example 8 are as follows: 13 13C NMR (400 MHz, DMSO) δ 188.33 (s), 171.23 (s), 167.23 (s), 152.58 (s), 147.78 (s), 146.07 (s), 142.94 (s), 136.25 (s), 127.86 (s), 71.46 (s), 62.89 (s), 57.34 (s), 55.27 (s), 51.39 (s), 46.45 (s), 43.78 (s), 33.54 (s), 31.29 (s), 29.53 (s), 24.75 (s), 21.63 (s), 21.45 (s), 19.34 (s), 16.25 (s), 15.46 (s), 11.25 (s).

[0134] The melting point of the supramolecular collagen-sodium hyaluronate eutectic (1:50, without nitrogen) prepared in Example 8 is 212 °C.

[0135] The two-dimensional nuclear magnetic resonance hydrogen spectrum of the supramolecular collagen-sodium hyaluronate eutectic (1:50, without nitrogen) prepared in Example 8 is as Figure 17 shown. It can be seen that there is no non-covalent interaction force between collagen and sodium hyaluronate, indicating that no supramolecular structure is formed between collagen and sodium hyaluronate.

[0136] (2) Transmission electron microscopy tests were carried out on the supramolecular collagen-sodium hyaluronate eutectics prepared in Examples 1-8.

[0137] The results are as Figure 18 shown. It can be seen that:

[0138] The particle size of the supramolecular collagen-sodium hyaluronate eutectic (10:1) prepared in Example 1 is about 18.2 nm and is evenly dispersed.

[0139] The particle size of the supramolecular collagen-sodium hyaluronate eutectic (1:1) prepared in Example 2 is about 486.2 nm and it is evenly dispersed.

[0140] The particle size of the supramolecular collagen-sodium hyaluronate eutectic (40:1) prepared in Example 3 is about 498.6 nm and it is evenly dispersed.

[0141] The particle size of the supramolecular collagen-sodium hyaluronate eutectic (10:1, without nitrogen) prepared in Example 4 is about 40.5 nm and it is evenly dispersed.

[0142] The particle size of the supramolecular collagen-sodium hyaluronate eutectic (1:1, without nitrogen) prepared in Example 5 is about 476.5 nm and it is evenly dispersed.

[0143] The particle size of the supramolecular collagen-sodium hyaluronate eutectic (40:1, without nitrogen) prepared in Example 6 is about 469.7 nm and it is evenly dispersed.

[0144] The particle size of the supramolecular collagen-sodium hyaluronate eutectic (1:50) prepared in Example 7 is about 310.4 nm and it is evenly dispersed.

[0145] The particle size of the supramolecular collagen-sodium hyaluronate eutectic (1:50, without nitrogen) prepared in Example 8 is about 335.7 nm and it is evenly dispersed.

[0146] (3) The Franz diffusion cell method was used to test the permeability of the supramolecular collagen-sodium hyaluronate eutectics prepared in Examples 1-8 with an artificial membrane.

[0147] The results are as Figure 19 shown (wherein, in the physical mixture of collagen + sodium hyaluronate, the molar ratio of collagen to sodium hyaluronate is 10:1), it can be seen that:

[0148] The permeability of the supramolecular collagen-sodium hyaluronate eutectic (10:1) prepared in Example 1 is 9.4 times that of collagen and 2.92 times that of sodium hyaluronate.

[0149] The permeability of the supramolecular collagen-sodium hyaluronate eutectic (1:1) prepared in Example 2 is 6.54 times that of collagen and 2.03 times that of sodium hyaluronate.

[0150] The permeability of the supramolecular collagen-sodium hyaluronate eutectic (40:1) prepared in Example 3 is 7.11 times that of collagen and 2.21 times that of sodium hyaluronate.

[0151] The permeability of the supramolecular collagen-sodium hyaluronate eutectic (10:1, without nitrogen) prepared in Example 4 is 7.78 times that of collagen and 2.42 times that of sodium hyaluronate.

[0152] The permeability of the molecular collagen-sodium hyaluronate eutectic (1:1, without nitrogen) prepared in Example 5 is 5.17 times that of collagen and 1.61 times that of sodium hyaluronate.

[0153] The permeability of the supramolecular collagen-sodium hyaluronate eutectic (40:1, without nitrogen) prepared in Example 6 is 5.40 times that of collagen and 1.67 times that of sodium hyaluronate.

[0154] The permeability of the supramolecular collagen-sodium hyaluronate eutectic (1:50) prepared in Example 7 is 4.93 times that of collagen and 1.53 times that of sodium hyaluronate.

[0155] The permeability of the supramolecular collagen-sodium hyaluronate eutectic (1:50, without nitrogen) prepared in Example 8 is 4.64 times that of collagen and 1.44 times that of sodium hyaluronate.

[0156] (4) Test the firming and anti-wrinkle effects of the supramolecular collagen-sodium hyaluronate eutectic (10:1) prepared in Example 1. The details are as follows:

[0157] Use in vitro skin tissue for culture, add the supramolecular collagen-sodium hyaluronate eutectic (10:1), after irradiation with UVA and UVB, add chondroitin sulfate, and detect its content by immunofluorescence. From Figure 20 It can be seen that when the molar ratio of collagen to sodium hyaluronate is 10:1, compared with the negative control group, when physically mixed collagen and sodium hyaluronate are added, the content of chondroitin sulfate increases, and the increase rate is 18.33%. When the supramolecular collagen-sodium hyaluronate eutectic (10:1) is added, the content of chondroitin sulfate increases significantly, and the increase rate is 45.00%. This shows that the supramolecular collagen-sodium hyaluronate eutectic (10:1) can increase the content of chondroitin sulfate and has a firming effect.

[0158] Use in vitro skin tissue for culture, add the supramolecular collagen-sodium hyaluronate eutectic (10:1), after irradiation with UVA and UVB, detect the content of Collagen-I by immunofluorescence. From Figure 21It can be seen that when the molar ratio of collagen to sodium hyaluronate is 10:1, compared with the negative control group, when physically mixed collagen and sodium hyaluronate are added, the content of Collagen-I increases, with a promotion rate of 144.83%. When the supramolecular collagen-sodium hyaluronate eutectic (10:1) is added, the content of Collagen-I increases significantly, with a promotion rate of 186.21%. This shows that the supramolecular collagen-sodium hyaluronate eutectic (10:1) can increase the content of Collagen-I and has anti-wrinkle effects.

[0159] In vitro skin tissues were cultured and the supramolecular collagen-sodium hyaluronate eutectic (10:1) was added. After irradiation with UVA and UVB, the content of Collagen-III was detected by immunofluorescence. Figure 22 It can be seen that when the molar ratio of collagen to sodium hyaluronate is 10:1, compared with the negative control group, when physically mixed collagen and sodium hyaluronate are added, the content of Collagen-III increases, with a promotion rate of 137.5%. When the supramolecular collagen-sodium hyaluronate eutectic (10:1) is added, the content of Collagen-III increases significantly, with a promotion rate of 156.25%. This shows that the supramolecular collagen-sodium hyaluronate eutectic (10:1) can increase the content of Collagen-III and has anti-wrinkle effects.

[0160] In vitro skin tissues were cultured and the supramolecular collagen-sodium hyaluronate eutectic (10:1) was added. After irradiation with UVA and UVB, it was stained with H&E, and the thickness of the epidermal viable cell layer was observed and recorded by taking pictures under a microscope. Figure 23 It can be seen that when the molar ratio of collagen to sodium hyaluronate is 10:1, based on in vitro skin tissues, compared with the negative control group, when physically mixed collagen and sodium hyaluronate are added, the thickness of the epidermal viable cell layer increases, with a promotion rate of 31.05%. When the supramolecular collagen-sodium hyaluronate eutectic (10:1) is added, the thickness of the epidermal viable cell layer increases significantly, with a promotion rate of 37.43%. This shows that the supramolecular collagen-sodium hyaluronate eutectic (10:1) can increase the thickness of the epidermal viable cell layer and has anti-wrinkle effects.

[0161] At room temperature, the optimal molar ratio of collagen to sodium hyaluronate during the preparation of the supramolecular collagen-sodium hyaluronate eutectic is 10:1, the optimal reaction temperature is 40 °C, the optimal reaction time is 24 h, and the optimal drying time is 48 h. A magnetic stirrer was used to mix evenly to obtain a mixed solution, and the system was observed to be completely dissolved to a clear state, and then the supramolecular collagen-sodium hyaluronate eutectic was obtained by freeze-drying.

[0162] The above results indicate that the supramolecular collagen-sodium hyaluronate eutectic has good biocompatibility and high bioavailability, and the supramolecular collagen-sodium hyaluronate eutectic improves the skin permeability of collagen.

[0163] In summary, in the present invention, collagen and sodium hyaluronate are used to form a supramolecular collagen-sodium hyaluronate solution through a eutectic reaction, and the product is separated and purified by concentration and crystallization, and the supramolecular collagen-sodium hyaluronate eutectic is obtained after drying. Compared with single collagen, the supramolecular collagen-sodium hyaluronate eutectic improves skin permeability and simultaneously has the efficacy and biological activities of both collagen and sodium hyaluronate. Moreover, in in vitro skin tissue culture, after adding the supramolecular collagen-sodium hyaluronate eutectic and irradiating with UVA and UVB, compared with the negative control group, the contents of chondroitin sulfate, Collagen-I, and Collagen-III increase, and the thickness of the viable epidermal cell layer increases, showing an anti-wrinkle effect. After being absorbed by the skin, the supramolecular collagen penetrates through the stratum corneum and binds to skin cells, participating in improving skin cell metabolism, which can increase collagen, thereby enhancing the skin tightness and making it firm and elastic.

[0164] It should be understood that the application of the present invention is not limited to the above examples. For those of ordinary skill in the art, improvements or modifications can be made according to the above description, and all such improvements and modifications should fall within the protection scope of the appended claims of the present invention.

Claims

1. A preparation method of a supramolecular collagen-sodium hyaluronate eutectic, characterized in that, Including the steps: Dissolve collagen and sodium hyaluronate in a solvent respectively to obtain a collagen solution and a sodium hyaluronate solution; Mix the collagen solution and the sodium hyaluronate solution, heat the mixed solution for a eutectic reaction to obtain a supramolecular collagen-sodium hyaluronate solution; Concentrate, recrystallize, filter and dry the supramolecular collagen-sodium hyaluronate solution to obtain a supramolecular eutectic constructed by collagen and sodium hyaluronate, that is, the supramolecular collagen-sodium hyaluronate eutectic.

2. The preparation method of the supramolecular collagen-sodium hyaluronate eutectic according to claim 1, characterized in that, The molar ratio of the collagen to the sodium hyaluronate is 1:40 to 40:

1.

3. The preparation method of the supramolecular collagen-sodium hyaluronate eutectic according to claim 2, wherein, The molar ratio of the collagen to the sodium hyaluronate is 1:1, 1:2, 1:3, 1:4, 1:5, 1:6, 1:7, 1:8, 1:9, 1:10, 1:20, 1:30, 1:40, 2:1, 3:1, 4:1, 5:1, 6:1, 7:1, 8:1, 9:1, 10:1, 20:1, 30:1 or 40:

1.

4. The preparation method of the supramolecular collagen-sodium hyaluronate eutectic according to claim 1, characterized in that, Heat the mixed solution for a eutectic reaction under an inert atmosphere.

5. The preparation method of the supramolecular collagen-sodium hyaluronate eutectic according to claim 1, wherein The temperature of the eutectic reaction is 30°C to 100°C, and the time of the eutectic reaction is 4h to 48h.

6. The preparation method of the supramolecular collagen-sodium hyaluronate eutectic according to claim 1, characterized in that, The recrystallization method is vacuum distillation; the drying method is freeze-drying, the temperature of the freeze-drying is -100°C to -30°C, and the time of the freeze-drying is 4h to 48h.

7. A supramolecular collagen-sodium hyaluronate eutectic, characterized in that, Prepared by the preparation method of the supramolecular collagen-sodium hyaluronate eutectic according to any one of claims 1 to 6; And / or, the supramolecular collagen-sodium hyaluronate eutectic is obtained by combining collagen and sodium hyaluronate through non-covalent bond forces; The particle size of the supramolecular collagen-sodium hyaluronate eutectic is 1nm to 500nm.

8. A composition, characterized in that, The composition includes the supramolecular collagen-sodium hyaluronate eutectic according to claim 7 as an active ingredient.

9. Use of a composition according to claim 8 in the preparation of a cosmetic, food, pharmaceutical or medical device product.

Images