Microneedle preparation based on polysulfonic acid mucopolysaccharide as well as preparation method and application of microneedle preparation

Abstract

The invention provides a multi-sulfonic acid mucopolysaccharide (MPS)-based microneedle preparation and a preparation method thereof. The microneedle preparation is prepared from polysulfonic mucopolysaccharide, hyaluronic acid, sodium carboxymethyl cellulose, glycerol, a pH regulator and water, has excellent stability and good transdermal drug delivery performance, and is suitable for phlebitis, varicosity, scar improvement and the like.

Description

Technical Field

[0001] The present invention relates to the technical field of pharmaceutical preparations, and particularly relates to a microneedle preparation based on mucopolysaccharide polysulfate (MPS), a preparation method thereof, and an application thereof. The microneedle preparation has excellent stability, solubility, and transdermal drug delivery performance, and is applicable to the treatment of diseases such as arthritis and varicose veins. Background Art

[0002] Mucopolysaccharide Polysulfate (MPS) is a naturally occurring glycosaminoglycan, a mixture of highly sulfated chondroitin sulfate-like polysaccharide molecules with a molecular weight between 5000 and 15000 daltons. The structural formula of mucopolysaccharide polysulfate is as follows:

[0003]

[0004] Mucopolysaccharide polysulfate widely exists in animal tissues, can promote the synthesis of mesenchymal cells and restore the ability of intercellular substances to retain moisture, and has various biological activities such as anti-inflammatory, antioxidant, moisturizing, anti-thrombosis formation, and promoting connective tissue regeneration. Research shows that MPS can promote fibroblast proliferation and collagen synthesis, and inhibit the activity of matrix metalloproteinases, thereby playing an anti-aging and scar repair-promoting role.

[0005] Currently, only cream preparations of mucopolysaccharide polysulfate have been developed and marketed (such as Hirudoid), and are clinically applied to the treatment of phlebitis with certain effects. However, its dosage form limitations are significant: 1) The semi-solid matrix results in a short residence time of the drug on the skin surface, and multiple daily administrations are required; 2) The cream dosage form has insufficient penetration rate into the dermis layer, and the drug is difficult to penetrate the skin cutin layer, affecting the efficacy of deep tissues; 3) Emulsifiers in the matrix may cause skin irritation reactions, and a considerable number of patients have contact dermatitis in clinical trials; 4) The cream drug is easily wiped off by clothes, has a short action time, large fluctuations in local drug concentration, and is difficult to release continuously, resulting in low patient compliance (frequent application is required). These defects limit the application of MPS in indications that require long-term administration such as chronic wound healing and scar repair. At the same time, the cream is easily rubbed off by close-fitting clothes during use, thus affecting the drug efficacy, and the patient compliance is poor. In addition, the cream is also easily contaminated and oxidized and deteriorated when exposed to the environment, and there are great inconveniences in clinical use.

[0006] In recent years, the research on new transdermal drug delivery systems has provided new ideas for breaking through the above limitations. Although technologies such as nanostructured lipid carriers and microneedle arrays can improve the transdermal efficiency, they are difficult to develop due to complex processes and high costs, and there is relatively little relevant research.

[0007] Microneedle technology is a novel transdermal drug delivery system that can penetrate the stratum corneum and directly deliver drugs to the dermis, thereby improving the bioavailability of drugs. As a new transdermal drug delivery method, the initially used microneedles were made of silicon and silicon dioxide. Although such microneedles have a mature preparation process, they have disadvantages such as poor biocompatibility, brittle texture, and slow production speed, which limit their industrialization. Another type is metal microneedles, which have good mechanical strength but have disadvantages such as small drug loading capacity, complex processes, high processing costs, and generation of medical waste, also restricting the industrial production and clinical application of this type of microneedles. The needles of the new soluble microneedles are formed by soluble or biodegradable excipients. The drug components are directly present in the needles by mixing with the excipients. After the needles penetrate the skin, the drugs distributed in the needles start to be released. Due to the small size of this type of microneedles (the operable space is within 1 mm 3 ), higher requirements are imposed on the prescription and process control for industrialization.

[0008] Heparinoid is a macromolecular drug. Due to reasons such as stability and processes, there is currently no heparinoid microneedle preparation developed and marketed on the market. Therefore, if the limitations can be broken through, developing an MPS microneedle preparation with high stability, good solubility, and excellent mechanical properties, and solving the problems of complex processes and reduced costs, can achieve efficient drug delivery and good patient compliance, which has important practical significance. Summary of the Invention

[0009] The purpose of the present invention is to provide a microneedle preparation based on heparinoid and its preparation method to solve the deficiencies of poor permeability and low bioavailability of MPS cream preparations in the prior art, and to overcome the problems of insufficient stability and poor solubility of heparinoid microneedle preparations.

[0010] The present invention provides a microneedle preparation based on heparinoid, characterized in that its prescription contains heparinoid, hyaluronic acid, sodium carboxymethylcellulose, plasticizer, pH regulator, and water.

[0011] For the microneedle preparation based on heparinoid provided by the present invention, characterized in that the components in the prescription are calculated by weight percentage, the amount of heparinoid is 0.5 - 1%, the amount of hyaluronic acid is 20 - 30%, the amount of sodium carboxymethylcellulose is 10 - 15%, the amount of plasticizer is 3 - 8%, and the amount of vitamin C is 0.05 - 0.2%. In addition, an appropriate amount of pH regulator is also contained, and its usage amount is to adjust the pH of the prescription solution to 6.0 - 7.0, and the rest is water. The water is distilled water, purified water, or water for injection. The pH regulator is selected from hydrochloric acid, sodium hydroxide, or a combination of the two. The hydrochloric acid is 0.1 mol / L hydrochloric acid solution, and among them, the sodium hydroxide is 0.1 mol / L sodium hydroxide solution.

[0012] The microneedle preparation described above is characterized in that the plasticizer is selected from glycerol and / or polyethylene glycol 400, preferably glycerol.

[0013] The researchers of the present invention found in experiments that hyaluronic acids with different molecular weights have an impact on the hardness of the microneedle body of the present invention. Adding an appropriate amount of low-molecular-weight hyaluronic acid is beneficial to improving the hardness of the microneedle body, and the hardness of the needle body is reflected in the piercing rate when the microneedle is used. When the proportion of low-molecular-weight hyaluronic acid in the amount of hyaluronic acid used exceeds 25%, compared with the technical solution using hyaluronic acid without low-molecular-weight hyaluronic acid, the hardness of the prepared microneedle body is significantly improved. However, it is not that the higher the content of low-molecular-weight hyaluronic acid, the better. As the proportion of low-molecular-weight hyaluronic acid increases, the hardness of the microneedle body will not continue to increase accordingly, but the toughness will decrease. The microneedles prepared with a high content of low-molecular-weight hyaluronic acid are more likely to be damaged, broken, etc. when subjected to external forces or skin surface friction, and are prone to break during the puncture process and are difficult to penetrate the skin, affecting the normal use of the microneedles. Therefore, when formulating the prescription, it is necessary to consider the drug loading, the stability of the microneedles, and the dissolution and release rates of the microneedles during use to balance the amounts of low-molecular-weight hyaluronic acid and high-molecular-weight hyaluronic acid in the hyaluronic acid.

[0014] In the microneedle preparation of the present invention, the hyaluronic acid is selected from low-molecular-weight hyaluronic acid, high-molecular-weight hyaluronic acid, or a mixture of the two in any proportion; preferably, the hyaluronic acid is a mixture of low-molecular-weight hyaluronic acid and high-molecular-weight hyaluronic acid, wherein the mass ratio of low-molecular-weight hyaluronic acid to high-molecular-weight hyaluronic acid is (0.25 - 4):1; more preferably, in the hyaluronic acid, the mass ratio of low-molecular-weight hyaluronic acid to high-molecular-weight hyaluronic acid is (0.5 - 2):1; even more preferably, in the hyaluronic acid, the mass ratio of low-molecular-weight hyaluronic acid to high-molecular-weight hyaluronic acid is 1:1.

[0015] In the present invention, the molecular weight of the low-molecular-weight hyaluronic acid is between 10KDa and 500KDa; the molecular weight of the high-molecular-weight hyaluronic acid is higher than 2000KDa.

[0016] The present invention also provides a preparation method of a microneedle preparation based on mucopolysaccharide polysulfate, or the preparation method of the microneedle preparation described above, comprising the following steps:

[0017] (1) Dissolution: Add hyaluronic acid and sodium carboxymethylcellulose to water to completely dissolve, add mucopolysaccharide polysulfate, and continue stirring until completely dissolved; add a plasticizer and vitamin C, and stir evenly to obtain a mixed solution;

[0018] (2) pH adjustment: Use a pH regulator to adjust the pH of the mixed solution to 6.0 - 7.0;

[0019] (3) Degassing: Place the mixed solution under vacuum for 30 - 45 minutes to remove air bubbles;

[0020] (4) Drying: Pour the degassed solution into a microneedle mold and let it stand for 30 minutes; then first place the mold at -20°C for 2 hours for pre-freezing, and then transfer it to a freeze dryer for freeze-drying at -50°C and 0.1 mbar for 24 hours;

[0021] (5) Sterilization: After demolding, place the microneedles under an ultraviolet lamp for 30 minutes for sterilization.

[0022] In the degassing operation in step (3) of the above-mentioned preparation method, it means placing the solution under vacuum and discharging the dissolved gas in the solution through negative pressure. Usually, it can be operated in two stages: Stage 1 (low-pressure start), slowly evacuate the vacuum to -0.05 MPa and maintain for 5 minutes (to avoid violent boiling of the solution). Stage 2 (deep degassing), gradually increase the vacuum degree to -0.09 to -0.1 MPa and maintain for 30 - 40 minutes (appropriately extend to 60 minutes for high-viscosity solutions). During the degassing process, bubbles should be continuously observed escaping from the surface of the solution. If no bubbles are generated, it can be determined that the degassing is complete.

[0023] Functions of the degassing operation: 1) Eliminate air bubbles and ensure the integrity of the microneedle structure. Air bubbles are easily introduced during the dissolution or mixing process. If they remain in the microneedle solution, it will cause voids or fractures in the microneedle body (decrease in mechanical strength) and deformation of the needle tip (reduction in puncture performance) after casting. 2) Improve the uniformity of drug distribution. Air bubbles will hinder the uniform dispersion of drug molecules, resulting in differences in drug content in the microneedle array. 3) Avoid cracking of microneedles during the drying process. Residual air bubbles expand when heated during drying. For the microneedles of the present invention, if degassing is not carried out, obvious cracks will appear on the surface of the prepared microneedles.

[0024] In the above-mentioned preparation method, the dissolution step in step (1) is obtained through careful design, experimental verification, and improvement. The dissolution step of the preparation method of the present invention is crucial for the solution stability and component compatibility. By changing the addition order of the raw and auxiliary materials, the researchers of the present invention observed phenomena such as caking, local agglomeration, reduced swelling efficiency, extended dissolution time, etc. Moreover, some raw and auxiliary materials may undergo enzymatic hydrolysis due to excessive local concentration, affecting the clinical efficacy. Therefore, it is necessary to control the dissolution step of the raw and auxiliary materials in the preparation method of the present invention.

[0025] In the preparation method described above, the water in step (1) can be added all at once, or 80-90% of the water can be used to dissolve hyaluronic acid and sodium carboxymethylcellulose first, and the remaining water can be divided into several portions. When necessary, it is used to wash the containers containing other raw and auxiliary materials respectively, and then the washing water is added to the mixed solution until all the water in the final prescription amount is added to the final mixed solution. It can be understood that these two ways of adding the solvent are basically equivalent. The operation of adding the remaining part of the solvent after step-by-step washing is a common operation for those skilled in the art. The purpose is to ensure that the raw and auxiliary materials can be transferred to the preparation solution as completely as possible, reduce the residue of the raw and auxiliary materials on the container wall, so as to ensure the accurate content of each component in the preparation, meet the requirements of the prescription design, and avoid deviation of the preparation quality caused by the loss of raw and auxiliary materials.

[0026] For the microneedle preparation of the present invention, a backing layer can also be added as a supporting substrate to prevent the microneedles from breaking during storage or use. The backing layer is generally made of materials such as polyester film and polycarbonate film. The operation of adding a backing layer to the microneedle preparation can select the following methods: (1) Clean or activate the surface of the backing layer. According to the size and shape of the microneedle preparation, cut the backing material into an appropriate size and shape to ensure that the backing material can completely cover the back of the microneedle preparation and has a certain edge margin for easy operation and fixation. (2) Uniformly coat an appropriate amount of binder (such as acrylate adhesive) on the back of the microneedle preparation or the bonding surface of the backing material. Align and bond the microneedle preparation coated with the binder with the backing material accurately and cure it. Check whether the tips of the microneedles are damaged or deformed. If necessary, perform corresponding repairs or adjustments.

[0027] The usage method of the microneedle preparation of the present invention: Clean the treatment area, tear off the protective film, and align the microneedle array with the treatment area; press with fingers for 10-15 seconds to ensure that the needle body penetrates, and maintain the application time until the microneedles dissolve or follow the doctor's advice.

[0028] The microneedle mold described in the present invention can be purchased as a commercial product or customized according to requirements. Microneedle molds of different shapes can be prepared, and the specifications include the microneedle height ranging from 0.05 mm to 2 mm, and the base diameter ranging from 0.05 mm to 0.5 mm. The mold shape can be circular, square or rectangular, etc.

[0029] The microneedles obtained by the present invention are arranged in an array, and the length of the microneedles can be between about 50 μm and 2 mm. In most cases, they are between about 200 μm and 1200 μm, and ideally between about 500 μm and 1000 μm. When the microneedles are used, they are applied to the skin. The microneedles can effectively puncture the physical barrier of the skin, break through the stratum corneum, and directly deliver the drug to the dermis layer through the pores.

[0030] The microneedles of the present invention can be rapidly dissolved and separated after piercing the skin and enter the subcutaneous layer for absorption, reducing irritation rapidly during drug administration.

[0031] The microneedle preparation based on mucopolysaccharide polysulfate provided by the present invention can be applied to skin indications such as vascular-related diseases (such as superficial phlebitis, varicose veins and related complications), skin repair and scar management (such as scar hyperplasia and fibrosis, localized scleroderma), inflammation and wound repair (such as local soft tissue contusion / hematoma).

[0032] Compared with the traditional mucopolysaccharide polysulfate cream, the microneedle preparation of the present invention improves the stability and antioxidant performance of mucopolysaccharide polysulfate through a specific formulation and preparation process. The microneedles have good hardness and toughness, and at the same time have good piercing rate and stability. High-efficiency transdermal drug delivery can be achieved without adding penetration enhancers, the drug is released rapidly, the bioavailability is significantly improved, the medication time of patients can be significantly shortened, and the compliance is better.

[0033] In the present invention, the term "comprising" or "including" can be open-ended, semi-closed and closed. In other words, the term also includes "substantially consisting of" or "consisting of".

[0034] In the present invention, "weight percentage" is equivalent to "mass percentage" (w / w or % m / m), which refers to the proportion of the mass of a certain component in a mixture to the total mass of the mixture, usually expressed as a percentage. Description of the Drawings

[0035] None. Detailed Embodiments

[0036] The technical solutions of the present invention will be further described below in conjunction with specific embodiments. It should be understood that these embodiments are only used to illustrate the present invention, and are only preferred embodiments of the present application, and are not intended to limit the scope of the present invention. Any modifications, equivalent substitutions or improvements made within the spirit and principle of the present application shall be included in the protection scope of the present application. The experimental methods without specific conditions noted in the following embodiments are usually carried out under conventional conditions or according to the conditions recommended by the manufacturer.

[0037] Example 1

[0038]

[0039]

[0040] (1) Dissolution: Dissolve hyaluronic acid and sodium carboxymethylcellulose in deionized water, stir until completely dissolved; add mucopolysaccharide polysulfate, continue to stir until completely dissolved; add glycerol and vitamin C, and stir evenly;

[0041] (2) pH adjustment: Adjust the pH of the mixed solution to 6.0 - 7.0 using 0.1 mol / L hydrochloric acid solution or 0.1 mol / L sodium hydroxide solution;

[0042] (3) Degassing: Place the mixed solution in a vacuum desiccator for 30 - 45 minutes to remove air bubbles;

[0043] (4) Drying: Pour the degassed solution into a microneedle mold and let it stand for 30 minutes; then first place the mold at -20°C for pre-freezing for 2 hours, and then transfer it to a freeze dryer and dry it at -50°C and 0.1 mbar for 24 hours;

[0044] (5) Sterilization: After demolding, place the microneedles under an ultraviolet lamp for sterilization for 30 minutes, and then package and store.

[0045] Example 2

[0046] Ingredient Proportion % (w / w) Mucopolysaccharide polysulfate 1 Hyaluronic acid (low molecular weight: high molecular weight = 1:2) 25 Sodium carboxymethyl cellulose 10 Glycerol 8 Vitamin C 0.2 0.1 mol / L hydrochloric acid solution Appropriate amount 0.1 mol / L sodium hydroxide solution Appropriate amount Distilled water Make up to 100%

[0047] (1) Dissolution: Dissolve hyaluronic acid and sodium carboxymethylcellulose in deionized water, stir until completely dissolved; add mucopolysaccharide polysulfate, and continue to stir until completely dissolved; add glycerol and vitamin C, and stir evenly;

[0048] (2) pH adjustment: Adjust the pH of the mixed solution to 6.0 - 7.0 using 0.1 mol / L hydrochloric acid solution or 0.1 mol / L sodium hydroxide solution;

[0049] (3) Degassing: Place the mixed solution in a vacuum desiccator for 30 - 45 minutes to remove air bubbles;

[0050] (4) Drying: Pour the degassed solution into a microneedle mold and let it stand for 30 minutes; then first place the mold at -20°C for pre-freezing for 2 hours, and then transfer it to a freeze dryer and dry it at -50°C and 0.1 mbar for 24 hours;

[0051] (5) Sterilization: After demolding, place the microneedles under an ultraviolet lamp for sterilization for 30 minutes, and then package and store.

[0052] Example 3

[0053] Ingredient Proportion % (w / w) Mucopolysaccharide polysulfate 1 Hyaluronic acid (low molecular weight: high molecular weight = 1:1) 25 Sodium carboxymethyl cellulose 10 Glycerol 3 Vitamin C 0.1 0.1 mol / L hydrochloric acid solution Appropriate amount 0.1 mol / L sodium hydroxide solution Appropriate amount Distilled water Make up to 100%

[0054] (1) Dissolution: Dissolve hyaluronic acid and sodium carboxymethylcellulose in deionized water, stir until completely dissolved; add mucopolysaccharide polysulfate, and continue to stir until completely dissolved; add glycerol and vitamin C, and stir evenly;

[0055] (2) pH adjustment: Adjust the pH of the mixed solution to 6.0 - 7.0 using 0.1 mol / L hydrochloric acid solution or 0.1 mol / L sodium hydroxide solution;

[0056] (3) Degassing: Place the mixed solution in a vacuum dryer for 30 - 45 minutes to remove air bubbles;

[0057] (4) Drying: Pour the degassed solution into a microneedle mold and let it stand for 30 minutes; then place the mold in a -20°C freezer for 2 hours, and then transfer it to a freeze dryer and dry it at -50°C and 0.1 mbar for 24 hours;

[0058] (5) Sterilization: After demolding, place the microneedles under an ultraviolet lamp for 30 minutes for sterilization, and then package and store them.

[0059] Example 4

[0060] Ingredient Proportion % (w / w) Mucopolysaccharide polysulfate 1 Hyaluronic acid (low molecular weight: high molecular weight = 2:1) 20 Sodium carboxymethyl cellulose 15 Glycerol 8 Vitamin C 0.2 0.1 mol / L hydrochloric acid solution Appropriate amount 0.1 mol / L sodium hydroxide solution Appropriate amount Distilled water Make up to 100%

[0061] (1) Dissolution: Dissolve hyaluronic acid and sodium carboxymethylcellulose in deionized water and stir until completely dissolved; add mucopolysaccharide polysulfate and continue to stir until completely dissolved; add glycerol and vitamin C and stir evenly;

[0062] (2) pH adjustment: Use 0.1 mol / L hydrochloric acid solution or 0.1 mol / L sodium hydroxide solution to adjust the pH of the mixed solution to 6.0 - 7.0;

[0063] (3) Degassing: Place the mixed solution in a vacuum dryer for 30 - 45 minutes to remove air bubbles;

[0064] (4) Drying: Pour the degassed solution into a microneedle mold and let it stand for 30 minutes; then place the mold in a -20°C freezer for 2 hours, and then transfer it to a freeze dryer and dry it at -50°C and 0.1 mbar for 24 hours;

[0065] (5) Sterilization: After demolding, place the microneedles under an ultraviolet lamp for 30 minutes for sterilization, and then package and store them.

[0066] Example 5

[0067] Ingredient Proportion % Mucopolysaccharide polysulfate 0.5 Hyaluronic acid (low molecular weight: high molecular weight = 4:1) 25 Sodium carboxymethyl cellulose 10 Polyethylene glycol 400 5 Vitamin C 0.2 0.1 mol / L hydrochloric acid solution Appropriate amount 0.1 mol / L sodium hydroxide solution Appropriate amount Distilled water Make up to 100%

[0068] (1) Dissolution: Dissolve hyaluronic acid and sodium carboxymethylcellulose in deionized water and stir until completely dissolved; add mucopolysaccharide polysulfate and continue to stir until completely dissolved; add polyethylene glycol 400 and vitamin C and stir evenly;

[0069] (2) pH adjustment: Use 0.1 mol / L hydrochloric acid solution or 0.1 mol / L sodium hydroxide solution to adjust the pH of the mixed solution to 6.0 - 7.0;

[0070] (3) Degassing: Place the mixed solution in a vacuum dryer for 30 - 45 minutes to remove air bubbles;

[0071] (4) Drying: Pour the degassed solution into the microneedle mold and let it stand for 30 minutes; then place the mold in a -20°C freezer for 2 hours, and then transfer it to a freeze dryer and dry it at -50°C and 0.1 mbar for 24 hours;

[0072] (5) Sterilization: After demolding, place the microneedles under an ultraviolet lamp for 30 minutes of sterilization, and then package and store them.

[0073] Comparative Example 1

[0074] Ingredient Proportion % (w / w) Mucopolysaccharide polysulfate 1 Low molecular weight hyaluronic acid 25 Sodium carboxymethyl cellulose 10 Glycerol 3 Vitamin C 0.1 0.1 mol / L hydrochloric acid solution Appropriate amount 0.1 mol / L sodium hydroxide solution Appropriate amount Distilled water Make up to 100%

[0075] (1) Dissolution: Dissolve hyaluronic acid and sodium carboxymethylcellulose in deionized water and stir until completely dissolved; add mucopolysaccharide polysulfate and continue to stir until completely dissolved; add glycerol and vitamin C and stir evenly;

[0076] (2) pH adjustment: Use 0.1 mol / L hydrochloric acid solution or 0.1 mol / L sodium hydroxide solution to adjust the pH of the mixed solution to 6.0 - 7.0;

[0077] (3) Degassing: Place the mixed solution in a vacuum dryer for 30 - 45 minutes of degassing to remove air bubbles;

[0078] (4) Drying: Pour the degassed solution into the microneedle mold and let it stand for 30 minutes; then place the mold in a -20°C freezer for 2 hours, and then transfer it to a freeze dryer and dry it at -50°C and 0.1 mbar for 24 hours;

[0079] (5) Sterilization: After demolding, place the microneedles under an ultraviolet lamp for 30 minutes of sterilization, and then package and store them.

[0080] Comparative Example 2

[0081] Ingredient Proportion % (w / w) Mucopolysaccharide polysulfate 1 High molecular weight hyaluronic acid 25 Sodium carboxymethyl cellulose 10 Glycerol 3 Vitamin C 0.1 0.1 mol / L hydrochloric acid solution Appropriate amount 0.1 mol / L sodium hydroxide solution Appropriate amount Distilled water Make up to 100%

[0082] (1) Dissolution: Dissolve hyaluronic acid and sodium carboxymethylcellulose in deionized water and stir until completely dissolved; add mucopolysaccharide polysulfate and continue to stir until completely dissolved; add glycerol and vitamin C and stir evenly;

[0083] (2) pH adjustment: Use 0.1 mol / L hydrochloric acid solution or 0.1 mol / L sodium hydroxide solution to adjust the pH of the mixed solution to 6.0 - 7.0;

[0084] (3) Degassing: Place the mixed solution in a vacuum dryer for 30 - 45 minutes of degassing to remove air bubbles;

[0085] (4) Drying: Pour the degassed solution into the microneedle mold and let it stand for 30 minutes; then place the mold at -20°C for pre-freezing for 2 hours, and then transfer it to a freeze dryer for drying at -50°C and 0.1 mbar for 24 hours;

[0086] (5) Sterilization: After demolding, place the microneedles under an ultraviolet lamp for sterilization for 30 minutes, and then package and store.

[0087] Comparative Example 3

[0088] Ingredient Proportion % (w / w) Mucopolysaccharide polysulfate 1 Hyaluronic acid (low molecular weight: high molecular weight = 1:1) 25 Sodium carboxymethyl cellulose 10 Glycerol 3 0.1 mol / L hydrochloric acid solution Appropriate amount 0.1 mol / L sodium hydroxide solution Appropriate amount Distilled water Make up to 100%

[0089] (1) Dissolution: Dissolve hyaluronic acid and sodium carboxymethylcellulose in deionized water and stir until completely dissolved; add mucopolysaccharide polysulfate and continue to stir until completely dissolved; add glycerol and vitamin C and stir evenly;

[0090] (2) pH adjustment: Use 0.1 mol / L hydrochloric acid solution or 0.1 mol / L sodium hydroxide solution to adjust the pH of the mixed solution to 6.0 - 7.0;

[0091] (3) Degassing: Place the mixed solution in a vacuum dryer for degassing for 30 - 45 minutes to remove air bubbles;

[0092] (4) Drying: Pour the degassed solution into the microneedle mold and let it stand for 30 minutes; then place the mold at -20°C for pre-freezing for 2 hours, and then transfer it to a freeze dryer for drying at -50°C and 0.1 mbar for 24 hours;

[0093] (5) Sterilization: After demolding, place the microneedles under an ultraviolet lamp for sterilization for 30 minutes, and then package and store.

[0094] Test Example 1: Test of the main drug release amount

[0095] Compare the mucopolysaccharide polysulfate microneedle preparation prepared according to Example 3 of the present invention with the commercially available mucopolysaccharide polysulfate cream:

[0096] Adopt the Franz diffusion cell method to simulate the skin penetration process, and respectively test the cumulative drug release amounts of the microneedle preparation and the cream within 8 hours.

[0097] Time (h) Release amount of microneedle preparation (%) Release amount of commercially available cream (%) 1 66.5 1.2 2 96.2 2.9 4 98.7 5.1 8 98.4 8.4

[0098] Compared with the cream preparation, due to its excellent solubility and transdermal performance, the microneedle preparation can rapidly release the drug after administration to improve the therapeutic effect, and compared with the cream, the administration time can be greatly shortened.

[0099] Test Example 2: Stability test

[0100] The microneedles and cream preparations prepared in Example 3 and Comparative Example 3 of the present invention were respectively stored under conditions of high temperature (40 °C), light (4500 Lux), and oxidation environment (0.5% hydrogen peroxide) for 3 months, and the residual rate of mucopolysaccharide polysulfate was measured.

[0101] Results:

[0102]

[0103] The accelerated stability effect of the microneedle preparation obtained by the preparation method of the present invention is significantly better than that of the cream preparation, and the stability of the microneedles added with an appropriate amount of vitamin C is better.

[0104] Morphological characterization of the microneedles in Test Example 3

[0105] The microneedles prepared according to Examples 1-5 were respectively placed flat on the object table, and the morphology of the microneedles was observed using a scanning electron microscope. The results showed that the prepared microneedles had a smooth surface, a neat array, and intact needle shapes.

[0106] Penetration rate test in Test Example 4

[0107] The microneedles prepared in Examples 1 to 5 and Comparative Examples 1 to 2 were used for skin penetration rate testing. The specific operation method was as follows: After pressing the microneedle patch on the depilated rat skin for 30 s, the microneedles were removed, 1% trypan blue was applied to the skin surface for 2 min and then wiped off, and the number of pinholes on the skin was observed and recorded under a microscope, and divided by the total number of needle bodies on the microneedle patch applied to the skin, that is, the penetration rate. The test was carried out in parallel 3 times, and the results are shown in the following table.

[0108]

[0109] It can be seen from the results that: in the microneedle formulation of the present invention, the penetration rate of Example 3 is the best. As the proportion of low-molecular-weight hyaluronic acid in hyaluronic acid increases, the microneedle penetration rate first increases and then decreases, indicating that adding a certain amount of low-molecular-weight sodium hyaluronate is beneficial to improving the hardness of the microneedle body of the present invention, but if the addition amount of low-molecular-weight sodium hyaluronate is too much, it will also cause the mechanical strength of the microneedle body to decrease, and when puncturing the skin, the microneedles may bend or break due to being unable to withstand the pressure, resulting in ineffective penetration of the stratum corneum.

Claims

1. A micro-needle preparation based on mucopolysaccharide polysulfate, characterized in that, Its prescription contains mucopolysaccharide polysulfate, hyaluronic acid, sodium carboxymethylcellulose, plasticizer, pH regulator and water.

2. The microneedle preparation according to claim 1, wherein, In terms of weight percentage, the components in the prescription are as follows: the amount of mucopolysaccharide polysulfate is 0.5 - 1%, the amount of hyaluronic acid is 20 - 30%, the amount of sodium carboxymethylcellulose is 10 - 15%, the amount of plasticizer is 3 - 8%, and the amount of vitamin C is 0.05 - 0.2%.

3. The microneedle preparation according to claim 1 or 2, characterized in that, The plasticizer selected is glycerol or polyethylene glycol 400.

4. The microneedle preparation according to claim 3, wherein The plasticizer is glycerol.

5. The microneedle preparation according to claim 1 or 2, characterized in that, The pH regulator is selected from hydrochloric acid, sodium hydroxide or a combination of both.

6. The microneedle preparation according to any one of claims 1-3, characterized in that, The hyaluronic acid is selected from low - molecular - weight hyaluronic acid, high - molecular - weight hyaluronic acid or a mixture of the two in any proportion.

7. The microneedle preparation according to claim 6, wherein, The hyaluronic acid is a mixture of low - molecular - weight hyaluronic acid and high - molecular - weight hyaluronic acid, and the mass ratio of low - molecular - weight hyaluronic acid to high - molecular - weight hyaluronic acid is (0.25 - 4):

1.

8. The microneedle preparation according to claim 7, wherein In the hyaluronic acid, the mass ratio of low - molecular - weight hyaluronic acid to high - molecular - weight hyaluronic acid is (0.5 - 2):

1.

9. The microneedle preparation according to claim 8, characterized in that, In the hyaluronic acid, the mass ratio of low - molecular - weight hyaluronic acid to high - molecular - weight hyaluronic acid is 1:

1.

10. The microneedle preparation according to any one of claims 1-9, characterized in that, The micro - needle preparation is prepared by the following steps: (1) Dissolution: Add hyaluronic acid and sodium carboxymethylcellulose to water and dissolve completely, then add mucopolysaccharide polysulfate, and continue stirring until completely dissolved; add plasticizer and vitamin C, and stir evenly to obtain a mixed solution; (2) pH adjustment: Use a pH regulator to adjust the pH of the mixed solution to 6.0 - 7.0; (3) Degassing: Place the mixed solution under vacuum for 30 - 45 minutes to remove air bubbles; (4) Drying: Pour the degassed solution into a micro - needle mold and let it stand for 30 minutes; then first place the mold at - 20°C for pre - freezing for 2 hours, and then transfer it to a freeze - dryer, and freeze - dry at - 50°C, under the condition of 0.1 mbar for 24 hours; (5) Sterilization: After demolding, place the micro - needles under an ultraviolet lamp for sterilization for 30 minutes.

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