Preparation method of cationic hyaluronic acid microspheres and the resulting products and applications

Abstract

The present invention discloses a method for preparing cationic hyaluronic acid microspheres, as well as the resulting products and applications. The method prepares hyaluronic acid polyester microspheres by a water / oil / water double emulsion solvent volatilization method, then introduces amino groups onto the microsphere surfaces through an aminolysis reaction to construct cationic hyaluronic acid microspheres. The microspheres are then applied to moisturizing finishing of textile fabrics. The cations of the microspheres and the anions on the surface of the fabric fibers form an electrostatic force, strengthening the bonding between the microspheres and the fabric, improving the washability of hyaluronic acid textiles, and resolving the problem of hyaluronic acid textiles being unable to retain moisture over the long term.

Description

Technical Field

[0001] The present invention relates to a cationic hyaluronic acid microsphere for a textile fabric finishing agent, and also relates to a preparation method of the cationic hyaluronic acid microsphere and application of the cationic hyaluronic acid microsphere in a textile fabric finishing agent, belonging to the technical field of textile finishing agents. Background Art

[0002] Currently, with rising consumption, functional textiles are gaining popularity, such as those with antibacterial, waterproof, insect-repellent, and skin-beautifying properties. In 2006, the UK's Textile and Apparel Standards Agency formally defined cosmetic textiles, stating that cosmetic active ingredients contained in textiles should be encapsulated in microcapsules before being introduced into the textile. Microencapsulation can also be used to encapsulate other active ingredients.

[0003] Hyaluronic acid, also known as hyaluronic acid, is an active polysaccharide widely used in skincare products. Its structure allows one hyaluronic acid molecule to bind to 1,000 water molecules, resulting in strong moisturizing properties. Adding hyaluronic acid to fabrics can maintain moisture in the skin, keeping it healthy and hydrated. In recent years, textile end-use products incorporating hyaluronic acid have gradually emerged, with products such as "hyaluronic acid face masks," "hyaluronic acid T-shirts," "hyaluronic acid eye masks," and "hyaluronic acid four-piece sets" now available on the market. However, hyaluronic acid textiles are still in their infancy, with no unified industry or national standards. The definition and content of hyaluronic acid textiles are also unclear. The group standard specifies that a hyaluronic acid content greater than 100 ppm is considered hyaluronic acid textiles. Currently, the main methods for incorporating hyaluronic acid into textiles are fiber-based, padding-based, and self-assembly-based.

[0004] The fiber method is to add hyaluronic acid to the spinning solution and then spin it to obtain hyaluronic acid fiber. This method has certain requirements on the amount of hyaluronic acid added. Too much addition will affect the fiber's stretching, regain and other properties. Too little addition will not be able to exert the skin care effect of hyaluronic acid, and the process requirements are relatively high.

[0005] The principle of the self-assembly method is that hyaluronic acid molecules contain a large amount of negative charges, and common fabric fibers also carry a large amount of negative charges. Therefore, macromolecules containing positive charges can be electrostatically self-assembled on the fiber surface, and then hyaluronic acid can be assembled on the fiber surface using the same principle to obtain fibers containing hyaluronic acid. This method has not yet been commercialized.

[0006] Currently, the most widely used method is padding. Hyaluronic acid is formulated into a finishing solution and then introduced into the fabric via padding. This method is simple, low-cost, and has little impact on the existing process. However, hyaluronic acid remains on the fabric surface through adsorption, lacking strong binding force and difficult to retain long-term. Furthermore, natural fibers are mostly anionic, and the large negative charge carried by hyaluronic acid also affects its binding to fabric fibers. Summary of the Invention

[0007] In view of the widespread problems in adding hyaluronic acid to textiles in the above industry, the present invention provides a cationic hyaluronic acid microsphere and a preparation method thereof. This method prepares hyaluronic acid into microspheres, which can achieve the sustained release of hyaluronic acid. At the same time, amino groups are introduced on the surface of the microspheres, thereby enhancing the binding ability between hyaluronic acid and fabric fibers, and further improving the wash resistance of hyaluronic acid in the fabric.

[0008] The specific technical solutions of the present invention are as follows:

[0009] A method for preparing cationic hyaluronic acid microspheres comprises the following steps:

[0010] (S1) preparing hyaluronic acid microspheres by a double emulsion solvent evaporation method;

[0011] (S2) performing surface aminolysis reaction on the obtained hyaluronic acid microspheres to obtain cationic hyaluronic acid microspheres.

[0012] Furthermore, in step (S1), the double emulsion solvent evaporation method is a water / oil / water (W / O / W) double emulsion solvent evaporation method, which is widely reported in the prior art. Hyaluronic acid microspheres can be prepared according to the methods disclosed in the prior art. During the preparation, hyaluronic acid microspheres are prepared using a hyaluronic acid substance as the core material and a polyester biodegradable material as the wall material.

[0013] Further, in step (S1), the hyaluronic acid substance is hyaluronic acid or / and its salt, and the hyaluronic acid substance can be one of hyaluronic acid or its salt, or can be two or more. The molecular weight of the hyaluronic acid substance is 10kDa-2000kDa, for example, 10kDa-30kDa, 30kDa-50kDa, 50kDa-80kDa, 80kDa-100kDa, 100kDa-300kDa, 300kDa-500kDa, 500kDa-800kDa, 800kDa-1000kDa, 1000kDa-1300kDa, 1300kDa-15 kDa, 1500 kDa to 1800 kDa, 1800 kDa to 2000 kDa, further, it can be 10 kDa, 50 kDa, 100 kDa, 200 kDa, 300 kDa, 400 kDa, 500 kDa, 600 kDa, 700 kDa, 800 kDa, 900 kDa, 1000 kDa, 1300 kDa, 1500 kDa, 1800 kDa, 2000 kDa.

[0014] Preferably, in step (S1), hyaluronic acid is a mixture of a low molecular weight hyaluronic acid substance with a molecular weight of 10kDa to 100kDa and a high molecular weight hyaluronic acid substance with a molecular weight of 1000kDa to 2000kDa, and the two are mixed and have better performance. Low molecular weight hyaluronic acid can penetrate into the skin surface layer when applied externally because of its relative molecular mass, play a role inside the skin, promote skin metabolism, remove oxygen free radicals, and make the skin soft, smooth and elastic. High molecular weight hyaluronic acid has good viscoelasticity, lubricity and moisture retention, can form a protective film on the skin surface, lock moisture, prevent water evaporation, and keep it moisturizing for a long time. The molecular weight of low molecular weight hyaluronic acid substances can be 10kDa-30kDa, 30kDa-50kDa, 50kDa-80kDa, 80kDa-100kDa, and the molecular weight of high molecular weight hyaluronic acid substances can be 1000kDa-1300kDa, 1300kDa-1500kDa, 1500kDa-1800kDa, 1800kDa-2000kDa.

[0015] Furthermore, when the hyaluronic acid substance is a mixture of a low molecular weight hyaluronic acid substance with a molecular weight of 10 kDa to 100 kDa and a high molecular weight hyaluronic acid substance with a molecular weight of 1000 kDa to 2000 kDa, there is no special requirement for the ratio of the two, and any ratio can be used. For example, the mass ratio of the low molecular weight hyaluronic acid substance to the high molecular weight hyaluronic acid substance can be 100:1, 80:1, 50:1, 30:1, 20:1, 10:1, 8:1, 6:1, 5:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:5, 1:6, 1:8, 1:10, 1:20, 1:30, 1:50, 1:80, 1:100, etc.

[0016] Furthermore, in step (S1), the polyester biodegradable material includes at least one of polymer carrier materials such as polylactic acid (PLA), polycaprolactone (PCL), polyglycolic acid (PGA), and polylactic acid-glycolic acid copolymer (PLGA).

[0017] Preferably, the polyester biodegradable material is polylactic acid or polylactic acid-glycolic acid copolymer. In the polylactic acid-glycolic acid copolymer, the content of monomer lactic acid in the total monomers is preferably 50-90wt%, for example, 50%, 60%, 70%, 80%, 90%, and the content of glycolic acid in the total monomers is preferably 10-50wt%, for example, 10%, 20%, 30%, 40%, 50%.

[0018] Furthermore, in step (S1), the molecular weight of the polyester biodegradable material is 5 kDa to 300 kDa, for example, 5 kDa, 10 kDa, 30 kDa, 50 kDa, 80 kDa, 100 kDa, 120 kDa, 150 kDa, 180 kDa, 200 kDa, 220 kDa, 240 kDa, 260 kDa, 280 kDa, and 300 kDa.

[0019] Furthermore, in step (S2), the amine used in the aminolysis reaction is a diamine compound, and the diamine compound has amino groups at both ends, for example, an alkyl diamine compound, and the molecular formula of the alkyl diamine compound is shown in the following formula (1), R 1 、R 2 、R 3 、R 4 Each of the alkyl groups is independently H or a C1-C6 alkyl group, and the value of n is 1 to 6. For example, the alkyldiamine compound can be ethylenediamine, hexamethylenediamine, etc.

[0020]

[0021] Furthermore, the above preparation method comprises the following specific steps:

[0022] (1) dissolving the polyester biodegradable material as a wall material in an organic solvent, and adding an emulsifier after the material is fully dissolved to obtain an oil phase solution;

[0023] (2) dissolving the core hyaluronic acid substance in water to obtain a solution containing the hyaluronic acid substance as the inner aqueous phase solution;

[0024] (3) dissolving the second emulsifier in water to obtain an emulsifier-containing aqueous phase solution as the external aqueous phase solution;

[0025] (4) adding the inner aqueous phase solution to the oil phase solution and homogenizing and emulsifying to form a W / O primary emulsion;

[0026] (5) adding the W / O primary emulsion to the external aqueous phase solution and stirring uniformly to obtain a W / O / W double emulsion;

[0027] (6) continuously stirring the emulsion to volatilize the organic solvent until the microspheres are solidified;

[0028] (7) Soaking the obtained microspheres in a diamine compound solution to perform an aminolysis reaction to obtain cationic hyaluronic acid microspheres.

[0029] Furthermore, in step (1), the mass volume percentage of the polyester biodegradable material is 2 to 15%, for example, 2%, 4%, 6%, 8%, 10%, 12%, and 15%, and the mass volume percentage of the emulsifier 1 is 0.1 to 10%, for example, 0.1%, 0.5%, 1%, 2%, 4%, 5%, 6%, 8%, and 10%.

[0030] Furthermore, in step (1), the purpose of the organic solvent is to fully dissolve the polyester biodegradable material, and it can be one, two or more of dichloromethane, acetone, ethyl acetate, chloroform, ether, etc.

[0031] Furthermore, in step (1), the purpose of the emulsifier is to fully emulsify the water phase and the oil phase. The emulsifier can be a conventional lipophilic emulsifier, such as a Span series lipophilic emulsifier, such as Span 80, etc. The performance of each Span series lipophilic emulsifier is similar.

[0032] Furthermore, in step (2), the mass volume percentage of the hyaluronic acid substance is 0.5-20%, for example, 0.5%, 1%, 1.5%, 2%, 4%, 6%, 8%, 10%, 12%, 14%, 16%, 18%, and 20%.

[0033] Furthermore, in step (2), when the hyaluronic acid substance contains a low molecular weight hyaluronic acid substance with a molecular weight of 10 kDa to 100 kDa, the mass volume percentage of the low molecular weight hyaluronic acid substance is 0.01% to 20%, for example, 0.01%, 0.1%, 0.5%, 1%, 1.5%, 2%, 4%, 6%, 8%, 10%, 12%, 14%, 16%, 18%, and 20%.

[0034] Furthermore, in step (2), when the hyaluronic acid substance contains a high molecular weight hyaluronic acid substance with a molecular weight of 1000 kDa to 2000 kDa, the mass volume percentage of the high molecular weight hyaluronic acid substance is 0.01% to 15%, for example, 0.01%, 0.05%, 0.1%, 0.3%, 0.5%, 0.8%, 1%, 2%, 4%, 6%, 8%, 10%, 12%, and 15%.

[0035] Furthermore, in step (3), the second emulsifier is a commonly used hydrophilic emulsifier, such as at least one of polyvinyl alcohol (PVA), Tweens, gelatin, poloxamer, and hydroxypropyl methylcellulose. The mass volume percentage of the emulsifier is 0.1-10%, such as 0.1%, 0.5%, 1%, 2%, 4%, 5%, 6%, 8%, and 10%.

[0036] Furthermore, in step (4), the volume ratio of the inner aqueous phase solution to the oil phase solution is 1:(3-20), for example, 1:3, 1:5, 1:8, 1:10, 1:12, 1:14, 1:15, 1:17, 1:20.

[0037] Furthermore, in step (4), the homogenization speed is 3000-20000 rpm, for example, 3000 rpm, 5000 rpm, 8000 rpm, 10000 rpm, 13000 rpm, 15000 rpm, 18000 rpm, 20000 rpm, and the homogenization time is 1-5 min.

[0038] Furthermore, in step (5), the volume ratio of the W / O primary emulsion to the external aqueous phase solution is 1:(5-20), for example, 1:5, 1:10, 1:15, or 1:20.

[0039] Furthermore, in step (5), the stirring rate is 200 to 3000 rpm, for example, 200 rpm, 500 rpm, 800 rpm, 1000 rpm, 1300 rpm, 1500 rpm, 1800 rpm, 2000 rpm, 2300 rpm, 2500 rpm, 2800 rpm, and 3000 rpm.

[0040] Furthermore, in step (6), the emulsion is continuously stirred to volatilize the organic solvent, thereby solidifying the microspheres. The stirring time is generally 4 to 8 hours.

[0041] Furthermore, in step (7), the diamine compound solution contains a diamine compound, a catalyst and a solvent, the mass volume percentage of the diamine compound is 1-20%, for example, 1%, 2%, 3%, 4%, 5%, 6%, 8%, 10%, 12%, 14%, 16%, 18%, 20%, and the mass volume percentage of the catalyst is 1-5%, for example, 1%, 2%, 3%, 4%, 5%.

[0042] Furthermore, in step (7), the catalyst is triethylamine, and the solvent of the diamine compound solution is DMSO, tetrahydrofuran, n-hexane, toluene, pyridine, etc.

[0043] Furthermore, in step (7), the microspheres are immersed in the diamine solution for 5-150 min, for example, 5 min, 10 min, 15 min, 20 min, 25 min, 30 min, 50 min, 70 min, 90 min, 100 min, 120 min, 140 min, 150 min, to carry out the aminolysis reaction, and the reaction temperature can be room temperature.

[0044] The cationic hyaluronic acid microspheres obtained according to the above method have a high encapsulation rate and good formability, contain amino groups on the surface, the average particle size of the microspheres is 50 to 100 μm, the hyaluronic acid content is 1 to 20%, and the encapsulation rate is 40 to 80%. The content of the hyaluronic acid in the microspheres can be regulated by controlling the concentration of the hyaluronic acid. The cationic hyaluronic acid microspheres of the present invention can be used in textile finishing agents to improve the binding force between the hyaluronic acid and the fiber, and enhance the retention rate of the hyaluronic acid in the textile after washing, thereby achieving a longer-lasting moisturizing and skin care effect.

[0045] The present invention also provides cationic hyaluronic acid microspheres, including surface-cationized W / O / W emulsion microspheres, wherein the inner aqueous phase contains hyaluronic acid substances, the microspheres have a particle size of 50 to 100 μm, a drug loading of 1 to 20%, and an encapsulation efficiency of 40 to 80%.

[0046] Furthermore, the hyaluronic acid substance in the inner aqueous phase is hyaluronic acid or / and its salt, and the molecular weight of the hyaluronic acid substance is selected from 10kDa-2000kDa; preferably, the hyaluronic acid substance is a mixture of a low molecular weight hyaluronic acid substance with a molecular weight of 10kDa-100kDa and a high molecular weight hyaluronic acid substance with a molecular weight of 1000kDa-2000kDa; the polyester biodegradable material in the oil phase includes at least one of polylactic acid, polycaprolactone, polyglycolic acid, and polylactic acid-glycolic acid copolymer, preferably polylactic acid or polylactic acid-glycolic acid copolymer, and preferably, the molecular weight of the polyester biodegradable material is 5kDa-300kDa.

[0047] Furthermore, the cationization of the cationized hyaluronic acid microspheres is to perform surface aminolysis reaction on the obtained hyaluronic acid microspheres. Preferably, the amine used in the aminolysis reaction is a diamine compound, more preferably an alkyl diamine compound represented by formula (1), wherein R1, R2, R3, and R4 are independently H or a C1-C6 alkyl group, and the value of n is 1-6;

[0048]

[0049] The present invention also provides a finishing agent, which is a textile finishing agent and contains the above-mentioned cationic hyaluronic acid microspheres.

[0050] Furthermore, the mass percentage of the cationic hyaluronic acid microspheres in the finishing agent is 1-20%, for example, 1%, 2%, 4%, 6%, 8%, 10%, 12%, 14%, 16%, 18%, 20%.

[0051] Furthermore, the finishing agent further comprises a surfactant, which is a cationic surfactant and / or a nonionic surfactant. The mass percentage of the surfactant in the finishing agent is 1-20%, for example, 1%, 2%, 4%, 6%, 8%, 10%, 12%, 14%, 16%, 18%, or 20%.

[0052] The present invention also provides application of the cationic hyaluronic acid microspheres or finishing agent in the textile field.

[0053] Compared with the prior art, the present invention has the following advantages and beneficial effects:

[0054] The hydrophilic properties of hyaluronic acid determine that hyaluronic acid textiles will inevitably lose a large amount of water after repeated washing. The present invention forms hyaluronic acid substances into microspheres and encapsulates the hyaluronic acid substances with wall materials, greatly improving the retention rate of hyaluronic acid substances in textiles after washing. At the same time, the present invention performs an aminolysis reaction on the surface of the microspheres, and the outer surface of the microspheres is coated with a large number of amino cations. These amino cations can form electrostatic forces with the anions on the fiber surface, thereby improving the binding force between the microspheres and the fibers. This not only solves the problem of the repulsive force between like charges between hyaluronic acid substances and fibers, but also increases the binding capacity of hyaluronic acid substances to fiber fabrics, solving the problem that traditional hyaluronic acid textiles cannot provide long-term moisturizing and skin care effects. BRIEF DESCRIPTION OF THE DRAWINGS

[0055] Figure 1 This is a microscopic morphology of the cationic hyaluronic acid microspheres in Example 1 of the present invention.

[0056] Figure 2 This is a microscopic morphology of the cationic hyaluronic acid microspheres in Comparative Example 4 of the present invention.

[0057] Figure 3 This is an electron microscope image of the bonding between the cationic hyaluronic acid microspheres and the fabric after the wash resistance test of Test Example 1 of the present invention. DETAILED DESCRIPTION

[0058] The present invention is further described below with reference to the examples. It should be understood that the examples are only used to further illustrate and explain the present invention and are not intended to limit the present invention.

[0059] Unless otherwise specified, the following concentrations are all percentages by mass and volume.

[0060] Example 1: Preparation of cationic hyaluronic acid microspheres

[0061] (1) Weigh 700 mg of PLA (200 kDa) and dissolve it in 10 mL of dichloromethane. After complete dissolution, add 50 μL of Span 80 to obtain an oil phase solution.

[0062] (2) Weigh 40 mg of sodium hyaluronate (a mixture of 10 mg of low molecular weight sodium hyaluronate with a molecular weight of 30 kDa to 50 kDa and 30 mg of high molecular weight sodium hyaluronate with a molecular weight of 1300 kDa to 1500 kDa) and dissolve it in 2 mL of ultrapure water to obtain an inner aqueous phase solution.

[0063] (3) The inner aqueous phase solution was added to the above oil phase and homogenized at 7000 rpm for 3 min to form a W / O primary emulsion.

[0064] (4) Weigh 700 mg of PVA and dissolve it in 70 mL of ultrapure water to obtain an external aqueous phase solution. Add the W / O primary emulsion to the external aqueous phase solution and stir magnetically at 800 rpm for 30 min to obtain a W / O / W double emulsion.

[0065] (5) The speed was slowed down to 300 rpm and the emulsion was stirred continuously for 4 h to evaporate the organic solvent until the microspheres hardened. The microspheres were then collected by centrifugation at 10,000 rpm for 10 min.

[0066] (6) The microspheres were immersed in a DMSO solution containing 8% by weight volume of ethylenediamine and 2% by weight volume of triethylamine for 10 minutes, and then washed with distilled water for 3 times to obtain surface-amino-modified hyaluronic acid polyester microspheres.

[0067] Example 2: Preparation of cationic hyaluronic acid microspheres

[0068] Surface-amino-modified hyaluronic acid polyester microspheres were prepared according to the method of Example 1, except that in step (1), 1500 mg of PLA (200 kDa) was used.

[0069] Example 3: Preparation of cationic hyaluronic acid microspheres

[0070] Surface-amino-modified hyaluronic acid polyester microspheres were prepared according to the method of Example 1, except that in step (1), 200 mg of PLA (200 kDa) was used.

[0071] Example 4: Preparation of cationic hyaluronic acid microspheres

[0072] Surface-amino-modified hyaluronic acid polyester microspheres were prepared according to the method of Example 1, except that in step (1), 700 mg of PLA (80 kDa) was used.

[0073] Example 5: Preparation of cationic hyaluronic acid microspheres

[0074] Surface-amino-modified hyaluronic acid polyester microspheres were prepared according to the method of Example 1, except that in step (1), 2100 mg of PLA (200 kDa) was weighed and dissolved in 30 mL of dichloromethane, and in step (4), 1850 mg of PVA was weighed and dissolved in 185 mL of ultrapure water. Thus, the volume ratio of the aqueous phase to the oil phase was 1:15.

[0075] Example 6: Preparation of cationic hyaluronic acid microspheres

[0076] Surface-amino-modified hyaluronic acid polyester microspheres were prepared according to the method of Example 1, except that in step (2), 400 mg of sodium hyaluronate (a mixture of 300 mg of low molecular weight sodium hyaluronate with a molecular weight of 30 kDa to 50 kDa and 100 mg of high molecular weight sodium hyaluronate with a molecular weight of 1300 kDa to 1500 kDa) was weighed and dissolved in 2 mL of ultrapure water to obtain an inner aqueous phase solution.

[0077] Example 7: Preparation of cationic hyaluronic acid microspheres

[0078] Surface-amino-modified hyaluronic acid polyester microspheres were prepared according to the method of Example 1, except that in step (2), 10 mg of sodium hyaluronate (10 mg of a mixture of high molecular weight sodium hyaluronates with a molecular weight of 1300 kDa to 1500 kDa) was weighed and dissolved in 2 mL of ultrapure water to obtain an inner aqueous phase solution.

[0079] Example 8: Preparation of cationic hyaluronic acid microspheres

[0080] Surface amino-modified hyaluronic acid polyester microspheres were prepared according to the method of Example 1, except that in step (4), 2400 mg of PVA was weighed and dissolved in 240 mL of ultrapure water, i.e., the ratio of primary emulsion to external aqueous phase was 1:20.

[0081] Example 9: Preparation of cationic hyaluronic acid microspheres

[0082] Surface amino-modified hyaluronic acid polyester microspheres were prepared according to the method of Example 1, except that: in step (1), the polyester biomaterial used was PLGA 50:50 .

[0083] Example 10: Preparation of cationic hyaluronic acid microspheres

[0084] Surface-amino-modified hyaluronic acid polyester microspheres were prepared according to the method of Example 1, except that in step (1), the polyester biomaterial used was PCL.

[0085] Example 11 Preparation of Cationic Hyaluronic Acid Microspheres

[0086] Surface-amino-modified hyaluronic acid polyester microspheres were prepared according to the method of Example 1, except that in step (1), the polyester biomaterial used was PGA.

[0087] Example 12 Preparation of Cationic Hyaluronic Acid Microspheres

[0088] Surface-amino-modified hyaluronic acid polyester microspheres were prepared according to the method of Example 1, except that in step (1), the organic solvent used was a mixed solution of dichloromethane and acetone in a volume ratio of 1:1.

[0089] Example 13 Preparation of Cationic Hyaluronic Acid Microspheres

[0090] Surface-amino-modified hyaluronic acid polyester microspheres were prepared according to the method of Example 1, except that in step (1), the organic solvent used was ethyl acetate.

[0091] Example 14 Preparation of Cationic Hyaluronic Acid Microspheres

[0092] Surface-amino-modified hyaluronic acid polyester microspheres were prepared according to the method of Example 1, except that in step (4), the external aqueous phase emulsifier used was gelatin.

[0093] Example 15 Preparation of Cationic Hyaluronic Acid Microspheres

[0094] Surface-amino-modified hyaluronic acid polyester microspheres were prepared according to the method of Example 1, except that in step (6), the concentration of ethylenediamine was 1% and the microspheres were immersed for 150 min.

[0095] Example 16 Preparation of Cationic Hyaluronic Acid Microspheres

[0096] Surface-amino-modified hyaluronic acid polyester microspheres were prepared according to the method of Example 1, except that in step (6), the concentration of ethylenediamine was 20% and the microspheres were immersed for 5 minutes.

[0097] Comparative Example 1: Preparation of cationic hyaluronic acid microspheres

[0098] Surface-amino-modified hyaluronic acid polyester microspheres were prepared according to the method of Example 1, except that in step (1), the mass of polylactic acid was 150 mg, that is, the mass volume concentration of polylactic acid in the oil phase solution was 1.5%.

[0099] Comparative Example 2: Preparation of cationic hyaluronic acid microspheres

[0100] Surface-amino-modified hyaluronic acid polyester microspheres were prepared according to the method of Example 1, except that in step (2), the mass of sodium hyaluronate used was 600 mg, and the molecular weight was 1300-1500 kDa, that is, the mass volume concentration of sodium hyaluronate was 30%.

[0101] Because the concentration of sodium hyaluronate is too high, it becomes gel-like when dissolved in water and does not mix evenly with the oil phase, making it impossible to form a stable primary emulsion.

[0102] Comparative Example 3: Preparation of cationic hyaluronic acid microspheres

[0103] Surface-amino-modified hyaluronic acid polyester microspheres were prepared according to the method of Example 1, except that: in step (2), 100 mg of sodium hyaluronate (a mixture of 25 mg of low molecular weight sodium hyaluronate with a molecular weight of 30 kDa to 50 kDa and 75 mg of high molecular weight sodium hyaluronate with a molecular weight of 1300 kDa to 1500 kDa) was weighed and dissolved in 5 mL of ultrapure water to obtain an inner aqueous phase solution. In step (4), 870 mg of PVA was weighed and dissolved in 87 mL of ultrapure water. Thus, the volume ratio of the inner aqueous phase solution to the oil phase solution was 1:2.

[0104] Comparative Example 4: Preparation of cationic hyaluronic acid microspheres

[0105] Surface-amino-modified hyaluronic acid polyester microspheres were prepared according to the method of Example 1, except that in step (6), the microspheres were immersed for 4 hours.

[0106] Comparative Example 5: Preparation of cationic hyaluronic acid microspheres

[0107] Surface-amino-modified hyaluronic acid polyester microspheres were prepared according to the method of Example 1, except that in step (1), 3500 mg of PLA (200 kDa) was dissolved in 50 mL of dichloromethane, and in step (4), 3020 mg of PVA was weighed and dissolved in 302 mL of ultrapure water, i.e., the volume ratio of the internal aqueous phase to the oil phase was 1:25.

[0108] Comparative Example 6: Preparation of cationic hyaluronic acid microspheres

[0109] Surface amino-modified hyaluronic acid polyester microspheres were prepared according to the method of Example 1, except that in step (4), 240 mg of PVA was weighed and dissolved in 24 mL of ultrapure water, i.e., the volume ratio of primary emulsion to external aqueous phase was 1:2.

[0110] Comparative Example 7: Preparation of cationic hyaluronic acid microspheres

[0111] Surface amino-modified hyaluronic acid polyester microspheres were prepared according to the method of Example 1, except that in step (4), 3600 mg of PVA was weighed and dissolved in 360 mL of ultrapure water, i.e., the volume ratio of primary emulsion to external aqueous phase was 1:30.

[0112] Performance Testing

[0113] 1. Particle size analysis of cationic hyaluronic acid microspheres

[0114] Take the dried microspheres, add an appropriate amount of distilled water to disperse them into a suspension, drop them onto a glass slide, observe the microsphere morphology under a microscope and count the particle size.

[0115] 2. Drug Loading and Encapsulation Efficiency of Cationic Hyaluronic Acid Microspheres

[0116] Drug loading of microspheres (%) = (mass of hyaluronic acid measured in microspheres / mass of microspheres) × 100%

[0117] Microsphere encapsulation efficiency (%) = (mass of hyaluronic acid measured in microspheres / total mass of hyaluronic acid input) × 100%

[0118] The hyaluronic acid quality determination method is as follows: the prepared microspheres are weighed and dissolved in dichloromethane. After the microspheres are fully dissolved, they are centrifuged at 5000 rpm for 10 minutes, the supernatant is discarded, and distilled water is added to the precipitate. After the precipitate is fully dissolved, the hyaluronic acid content in the solution is determined using the QB / T 4416-2012 standard.

[0119] The results of the determination of the average particle size, drug loading and encapsulation efficiency of the cationic hyaluronic acid microspheres in Examples 1-16 of the present invention and Comparative Examples 1, 3, 4, 5, 6 and 7 are shown in Table 1:

[0120] Table 1: Determination results of average particle size, drug loading and encapsulation efficiency of cationic hyaluronic acid microspheres

[0121]

[0122]

[0123] As shown in Table 1, the particle size of the cationized hyaluronic acid microspheres provided in Examples 1 to 16 of the present invention is 60 to 90 μm, the drug loading is 1.2 to 20%, and the encapsulation efficiency is 45 to 80%. In Comparative Example 1, as the wall material concentration decreases, the solution viscosity becomes lower, and the droplet volume produced is relatively small. Therefore, the microspheres prepared by the low concentration of wall material have a smaller particle size. The microspheres with a small particle size have a large specific surface area, and the contact area with the external aqueous phase becomes larger. The components in the microspheres are more easily diffused into the external aqueous phase, resulting in a decrease in the encapsulation efficiency. In Comparative Example 3, the volume ratio of the internal aqueous phase to the oil phase is increased, and the water droplets in the microspheres increase, resulting in an increase in the microsphere particle size. During the microsphere curing process, the organic solvent removal rate is accelerated, and the evaporation of the droplets inside the microspheres during the drying process will result in an increase in the number of microsphere holes, thereby increasing the rupture rate and reducing the encapsulation amount. In Comparative Example 4, the time of the aminolysis reaction was extended, and the polyester compound of the microsphere wall material gradually degraded and fell off, the microsphere weight was lost, the mechanical strength was reduced, and the core material hyaluronic acid was lost, thereby reducing the drug loading and encapsulation efficiency. In Comparative Example 5, when the water-oil volume ratio was too small, the emulsion droplets formed were uneven in size, the hollow rate of the obtained microspheres became high, and the drug loading and encapsulation efficiency were reduced. In Comparative Examples 6 and 7, the microsphere encapsulation efficiency was relatively low. The low volume of the external aqueous phase made the water layer thinner, increased the aggregation between the internal and external aqueous phases, and the high volume would lead to a decrease in the emulsion dispersion efficiency and an increase in the particle size of the emulsion droplets, all of which would cause the emulsion stability to decrease.

[0124] 3. The amino content of cationic hyaluronic acid microspheres was determined by the ninhydrin method, and the morphology of the microspheres was observed under a microscope.

[0125] The results of the determination of the surface amino content of the cationic hyaluronic acid microspheres in Examples 1-16 of the present invention and Comparative Examples 1, 3, 4, 5, 6, and 7 are shown in Table 2:

[0126] Table 2: Surface amino content of cationic hyaluronic acid microspheres

[0127] <![CDATA[Amino content (10 -7 mol / mg)]]> Example 1 2.73 Example 2 2.96 Example 3 2.11 Example 4 2.31 Example 5 2.89 Example 6 2.22 Example 7 2.75 Example 8 2.39 Example 9 2.73 Example 10 2.65 Example 11 2.62 Example 12 2.72 Example 13 2.43 Example 14 2.70 Example 15 2.97 Example 16 2.86 Comparative Example 1 1.32 Comparative Example 3 0.97 Comparative Example 4 2.74 Comparative Example 5 2.43 Comparative Example 6 1.32 Comparative Example 7 1.77

[0128] The aminolysis reaction is essentially a degradation reaction. One of the amino groups in the diamine forms an amide bond with the broken ester bond of the polyester polymer, and the free amino group at the other end is introduced to the surface of the microspheres. As shown in Table 2, the aminolysis reaction of the microspheres in the diamine compound solution increases the amino content on the surface. Figure 1 As shown in the figure, after 10 minutes of aminolysis reaction, it can be observed under the microscope that some microspheres are no longer round, and most microspheres are quasi-circular, and there are a large number of protrusions or grooves on the surface. This is caused by local degradation of the microsphere surface during the aminolysis reaction. The reduction of polymer concentration in Comparative Example 1 or the increase of the volume of the internal aqueous phase in Comparative Example 3 will cause the microsphere capsule wall to become thinner and the strength of the wall to decrease. The aminolysis reaction causes the capsule wall to decompose, thereby reducing the content of free amino groups on the surface of the microspheres. In Comparative Example 4, as the aminolysis time increases, the content of free amino groups on the surface of the microspheres increases, but at the same time, the microsphere matrix will gradually degrade, as shown in the figure. Figure 2 As shown, after 4 hours of aminolysis reaction, the microsphere structure became incomplete, with many fragments falling off the microsphere surface. This resulted in a low free amino group content on the microsphere surface, as well as low drug loading and encapsulation efficiency. Therefore, while increasing the amino group content on the microsphere surface, the aminolysis reaction time should be shortened as much as possible. The composite emulsions in Comparative Examples 6 and 7 showed low stability, resulting in an increased rupture rate of the resulting microspheres and a corresponding decrease in amino group content.

[0129] Test Example 1: Preparation of a finishing agent containing cationic hyaluronic acid microspheres and fabric finishing properties

[0130] Experimental group: 15 parts of cationic hyaluronic acid microspheres obtained in Example 1, 5 parts of undecylenic acid monoglyceride, 5 parts of fatty alcohol polyoxyethylene ether, 4 parts of cationic surfactant, and 71 parts of water were taken by weight, and the components were thoroughly mixed to obtain a finishing agent for later use.

[0131] Control group 1: The cationic hyaluronic acid microspheres were replaced with the hyaluronic acid microspheres obtained in step (5) of Example 1 that had not been subjected to the aminolysis reaction.

[0132] Control group 2: The cationic hyaluronic acid microspheres were replaced with a mixture solution of low molecular weight sodium hyaluronate with a molecular weight of 30 kDa to 50 kDa and high molecular weight sodium hyaluronate with a molecular weight of 1300 kDa to 1500 kDa in step (2) of Example 1.

[0133] The finishing agents described in the experimental and control groups 1 and 2 were diluted with water to a 5% working solution. This working solution was then added to a padding tank, and the fabric was completely immersed in the tank. The fabric was then dehydrated by rollers. The padding fabric was pre-dried at 80°C and then dried at 100°C to obtain a fabric containing hyaluronic acid.

[0134] (1) Hyaluronic acid washability test:

[0135] Fabrics containing hyaluronic acid were washed according to the washing method specified in GB / T 8629-2017, "Textile Testing - Household Washing and Drying Procedure." The binding between the washed fabric and the cationic hyaluronic acid microspheres was observed using a scanning electron microscope, and the hyaluronic acid content in the washed fabric was determined.

[0136] The hyaluronic acid content in fabrics is determined as follows: accurately weigh washed fabric, soak it in a dichloromethane solution, and ultrasonically extract it at 40°C three times for one hour each time. Combine the resulting extracts, separate the lower layer using a separatory funnel, and test the hyaluronic acid content in the fabric according to QB / T 4416-2012.

[0137] Table 3 Detection of hyaluronic acid content in fabrics

[0138]

[0139]

[0140] Electron microscope images of fabrics after washing Figure 3 As shown in the figure, it was observed that after washing 10 times, the microspheres were still successfully loaded on the fabric and the structure was intact. As can be seen from Table 3, the fabric treated with the cationic hyaluronic acid microspheres prepared by the present invention still had a hyaluronic acid content of more than 100ppm after washing 10 times, indicating that the cationic hyaluronic acid microspheres were firmly bonded to the fabric and had good water-washing resistance. In the control group 1, the microspheres were not subjected to amino treatment and were combined with the fabric by adsorption, resulting in a weak binding force. In the control group 2, the sodium hyaluronate was not embedded and dissolved in water and lost in large quantities, resulting in poor wash resistance.

[0141] (2) Determination of moisturizing efficacy of human skin:

[0142] After washing, the three groups of fabrics were subjected to human skin moisturizing tests to determine the moisturizing effects of the different groups of fabrics after washing.

[0143] Thirty healthy subjects were recruited and four 4*4cm marks were made on the flexor side of the left and right forearms. 2 Test area, cut the fabrics with different treatments into 4*4cm 2Apply the sample in different sizes to the test area. Initial stratum corneum moisture content was measured in each test area. The stratum corneum moisture content was then measured again 1, 3, and 8 hours after application. Taking the initial moisture content as 100%, the stratum corneum moisture content after different application times was calculated and the average moisture content for each group was calculated.

[0144] Table 4: Effects of different fabric groups on skin moisture content (%)

[0145] Before application 1 hour after application 3 hours after application 8 hours after application Experimental group 100 105 110 113 Control group 1 100 102 103 105 Control group 2 100 104 103 103

[0146] As shown in Table 4, compared with the other two groups, the experimental group had the highest water content in the stratum corneum within 8 hours, indicating that the cationic hyaluronic acid microspheres were more firmly bonded to the fabric, and that the hyaluronic acid was slowly released through the pores of the microspheres or the degradation of the microsphere walls, effectively retaining and releasing the hyaluronic acid molecules and exerting their excellent water-locking and moisturizing effects.

Claims

1. A method for preparing cationic hyaluronic acid microspheres, characterized in that: The following steps are involved: (1) Dissolving the polyester biodegradable material as the wall material in an organic solvent, and adding an emulsifier after the material is fully dissolved to obtain an oil phase solution; (2) dissolving the core hyaluronic acid substance in water to obtain a solution containing the hyaluronic acid substance as the inner aqueous phase solution; (3) dissolving the second emulsifier in water to obtain an aqueous phase solution containing the emulsifier as the external aqueous phase solution; (4) Add the inner aqueous phase solution to the oil phase solution and homogenize and emulsify to form a W / O primary emulsion; (5) Add the W / O primary emulsion to the external aqueous phase solution and stir evenly to obtain a W / O / W double emulsion; (6) Continuously stirring the emulsion to evaporate the organic solvent until the microspheres solidify; (7) Soaking the obtained microspheres in a diamine compound solution to perform an aminolysis reaction to obtain cationic hyaluronic acid microspheres; The diamine compound is an alkyl diamine compound represented by the following formula (1), wherein R 1 、R 2 、R 3 、R 4 are independently H or C1-C6 alkyl, and the value of n is 1-6; 。 2. The preparation method according to claim 1, characterized in that The hyaluronic acid substance is hyaluronic acid or / and its salt, and the molecular weight of the hyaluronic acid substance is selected from 10kDa-2000kDa; the polyester biodegradable material includes at least one of polylactic acid, polycaprolactone, polyglycolic acid, and polylactic acid-glycolic acid copolymer.

3. The preparation method according to claim 2, characterized in that The hyaluronic acid substance is a mixture of a low molecular weight hyaluronic acid substance with a molecular weight of 10 kDa to 100 kDa and a high molecular weight hyaluronic acid substance with a molecular weight of 1000 kDa to 2000 kDa.

4. The preparation method according to claim 2, characterized in that The polyester biodegradable material is polylactic acid or polylactic acid-glycolic acid copolymer.

5. The preparation method according to claim 1, 2 or 4, characterized in that: The molecular weight of polyester biodegradable materials is 5kDa~300kDa.

6. The preparation method according to claim 1, characterized in that In step (1), the mass volume percentage of the polyester biodegradable material is 2-15%; in step (2), the mass volume percentage of the hyaluronic acid substance is 0.5-20%; in step (4), the volume ratio of the inner aqueous phase solution to the oil phase solution is 1:3-20; in step (5), the volume ratio of the W / O primary emulsion to the outer aqueous phase solution is 1:5-20; in step (1), the emulsifier 1 is a lipophilic emulsifier; in step (3), the emulsifier 2 is a hydrophilic emulsifier; in step (7), the diamine compound solution contains a diamine compound and a catalyst, and the catalyst is triethylamine.

7. The preparation method according to claim 1 or 6, characterized in that In step (7), the microspheres are immersed in the diamine compound solution for 5-150 minutes to undergo an aminolysis reaction.

8. Cationized hyaluronic acid microspheres prepared by the method for preparing cationized hyaluronic acid microspheres according to any one of claims 1 to 7.

9. A cationic hyaluronic acid microsphere, characterized in that: The invention comprises W / O / W double emulsion microspheres with surface cationization, wherein the inner water phase contains hyaluronic acid substances, the particle size of the microspheres is 50-100 μm, the drug loading amount is 1-20%, and the encapsulation efficiency is 40-80%.

10. The cationic hyaluronic acid microspheres according to claim 9, characterized in that The hyaluronic acid substance in the inner aqueous phase is hyaluronic acid or / and its salt, and the molecular weight of the hyaluronic acid substance is selected from 10kDa-2000kDa.

11. The cationic hyaluronic acid microspheres according to claim 9 or 10, characterized in that: The hyaluronic acid substance is a mixture of a low molecular weight hyaluronic acid substance with a molecular weight of 10 kDa to 100 kDa and a high molecular weight hyaluronic acid substance with a molecular weight of 1000 kDa to 2000 kDa.

12. A finishing agent, characterized in that: Containing the cationic hyaluronic acid microspheres according to claim 8, 9, 10 or 11.

13. The finishing agent according to claim 12, characterized in that The finishing agent is a textile finishing agent.

14. Use of the cationic hyaluronic acid microspheres according to claim 8, 9, 10 or 11 or the finishing agent according to claim 12 or 13 in the field of textiles.

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