A composition microneedle for treating gout and a preparation method and application thereof

Abstract

The application discloses a composition microneedle for treating gout and a preparation method and application thereof. The application prepares a slow-release layer solution containing dextro-borneol, luteolin and a PEG modified uricase preparation, a burst release layer solution for shaping and a backing layer solution; a needle body connected by the slow-release layer and the burst release layer is obtained through a mold, the slow-release layer, the burst release layer and the backing layer are sequentially connected, and the composition microneedle for treating gout is formed. The composition microneedle can be quickly and efficiently absorbed through the skin, can be directly pasted on a disease site through a microneedle patch, and is simple and convenient to use; through an active separation type freezing process, active substances can be better released at a fixed point, a targeted treatment effect is achieved, a drug utilization rate is higher, and the composition microneedle has a wide application prospect in the field of treating gout.

Description

Technical Field

[0001] This invention belongs to the field of gout treatment technology, specifically relating to a microneedle composition for treating gout, its preparation method, and its application. Background Technology

[0002] Currently, with the rapid development of the national economy and the continuous improvement of people's living standards, more and more people are suffering from acute and chronic hyperuricemia, which has seriously affected the health of the nation. Gout is a crystal-related arthropathy caused by the deposition of monosodium urate crystals. Its onset is directly related to hyperuricemia caused by purine metabolism disorders and / or reduced uric acid excretion, and it is a metabolic rheumatic disease.

[0003] Based on the pathogenesis of gout, lowering serum uric acid levels, inhibiting uric acid production, and reducing inflammatory responses are considered effective treatment strategies. Currently, gout treatment typically involves traditional oral medications. However, the first-pass effect and gastrointestinal inactivation result in long drug durations and slow onset of action, preventing rapid delivery to the administration site. Furthermore, there are issues such as limited drug action, potential side effects on organs, and low transdermal absorption efficiency. In China, the treatment and prevention of hyperuricemia mainly rely on first-line chemical drugs such as allopurinol or imported recombinant uricase preparations.

[0004] Currently, there are patented treatments for gout using microneedle delivery, but the drugs used vary and are often singular, addressing only a single target. No patent reports have been found that simultaneously address gout from multiple dimensions—promoting uric acid dissolution and excretion, inhibiting uric acid production, and reducing inflammatory responses. Summary of the Invention

[0005] The primary objective of this invention is to overcome the shortcomings and deficiencies of the prior art and provide a method for preparing a compositional microneedle for treating gout.

[0006] Another object of the present invention is to provide a compositional microneedle for treating gout obtained by the above method.

[0007] Another object of the present invention is to provide the application of the above-described composition for treating gout using microneedles.

[0008] To achieve the above objectives, the present invention provides the following technical solution:

[0009] A method for preparing a compositional microneedle for treating gout, comprising the following steps:

[0010] S1. Preparation of dextrorotatory borneol microemulsion: The oil phase, surfactant, co-surfactant and dextrorotatory borneol are mixed and homogeneous to obtain solution A; under stirring, water droplets are added to the solution, and stirring is continued after the water droplets are added. After standing, dextrorotatory borneol microemulsion is obtained; wherein, the content of each component by mass percentage is as follows: dextrorotatory borneol 3-3.5%, oil phase 4-8%, surfactant 18-22%, co-surfactant 10-18%, water balance;

[0011] S2. Preparation of luteolin-dextrin microemulsion solution: Mix luteolin and the dextrin microemulsion obtained in step S1 evenly, let stand, and obtain luteolin-dextrin microemulsion solution; wherein, the content of luteolin in the luteolin-dextrin microemulsion solution is 3-3.5% by mass.

[0012] S3. Preparation of sustained-release layer solution: Mix uricase, the luteolin-dextrin microemulsion solution prepared in step S2, and the aqueous solution of needle excipient to obtain a sustained-release layer solution; wherein, the amounts of each component are as follows by mass percentage: uricase 5-10%, luteolin-dextrin microemulsion solution 10-15%, needle excipient 20-30%, and water as the balance;

[0013] S4. Preparation of burst-release layer solution: Mix the soluble polymer with water evenly to obtain a burst-release layer solution; wherein the concentration of the soluble polymer in the burst-release layer solution is 20-30% by mass.

[0014] S5. Preparation of backing layer solution: Mix soluble polymer, plasticizer and water evenly to obtain backing layer solution; wherein, the content of each component by mass percentage is as follows: soluble polymer 25-35%, plasticizer 1-2%, water as balance;

[0015] S6. Preparation of microneedles:

[0016] A. Add the sustained-release layer solution prepared in step S3 into the microneedle anion membrane to fill the microneedle anion membrane, scrape off the excess liquid, and dry and solidify to form a microneedle sustained-release layer;

[0017] B. Add the burst-release layer solution prepared in step S4 to the microneedle sustained-release layer in step A, so that it fills the microneedle anion membrane, scrape off the excess liquid, and dry and solidify to form the microneedle burst-release layer.

[0018] C. Spread the backing layer solution prepared in step S5 onto the microneedle release layer obtained in step B, and dry the entire microneedle to obtain the composition microneedle for treating gout.

[0019] The oil phase mentioned in step S1 is selected from polyethylene glycol glycerol octanoate and caprylic acid decanoate.

[0020] The surfactant mentioned in step S1 is selected from at least one of Tween 20 and Tween 40.

[0021] The co-surfactant mentioned in step S1 is selected from at least one of anhydrous ethanol and 1,2-propanediol.

[0022] The oil phase, surfactant, and co-surfactant mentioned in step S1 are preferably mixed in a mass ratio of 2:5.5 to 9:2.5 to 9.

[0023] The preferred method of mixing in step S1 is ultrasound.

[0024] The ultrasound duration is preferably 20-40 minutes; more preferably 30 minutes.

[0025] The temperature of the ultrasound is preferably 30-40℃; more preferably 35℃.

[0026] The ultrasonic power is preferably 250W; the ultrasonic frequency is preferably 40KHz.

[0027] The stirring speed in step S1 is preferably 200-300 rpm; more preferably 200-250 rpm.

[0028] The preferred stirring time in step S1 is 3 to 8 minutes; more preferably 5 minutes.

[0029] The settling time in step S1 is preferably 8 to 15 minutes; more preferably 10 minutes.

[0030] In step S2, luteolin and dextrorotatory borneol are mixed in a mass ratio of 1:1 in the luteolin-dextrin microemulsion solution.

[0031] The preferred method for uniform mixing in step S2 is to stir at 200-300 rpm for 3-8 minutes and then let it stand for 8-15 minutes; more preferably, to stir at 250 rpm for 5 minutes and then let it stand for 10 minutes.

[0032] The uricase mentioned in step S3 is preferably a PEG-modified uricase.

[0033] The uricase is preferably the humanized human-pig chimeric uricase mutant claimed in claim 1 of patent application CN202211051614.3.

[0034] The needle-forming material mentioned in step S3 is preferably methacryloyl hyaluronic acid (HAMA).

[0035] The aqueous solution of the needle-forming material mentioned in step S3 is preferably prepared by the following steps: under heating conditions, the needle-forming material is dissolved in water to obtain the aqueous solution of the needle-forming material.

[0036] The preferred heating conditions are stirring in a water bath at 70–80°C.

[0037] The soluble polymer mentioned in step S4 is preferably at least one of polyvinylpyrrolidone and polyvinyl alcohol.

[0038] The polyvinylpyrrolidone is preferably at least one of polyvinylpyrrolidone K30 and polyvinylpyrrolidone K90.

[0039] The polyvinyl alcohol is preferably polyvinyl alcohol 1788.

[0040] The mixing conditions described in step S4 are preferably stirring in a water bath at 70–80°C.

[0041] The soluble polymer mentioned in step S5 is preferably at least one of polyvinylpyrrolidone, polyvinyl alcohol, and chitosan; more preferably, it is a composition formed by polyvinyl alcohol and chitosan with polyvinylpyrrolidone.

[0042] The plasticizer mentioned in step S5 is preferably glycerin.

[0043] The mixing conditions described in step S5 are preferably those of stirring in a water bath at 70–80°C.

[0044] The operation of filling the microneedle anion membrane in step S6A is preferably a pressurized vacuum treatment; more preferably, it is pressurized in an environment of -80 to -100 for 15 to 20 minutes.

[0045] The drying conditions described in step S6A are preferably drying at a temperature of 2-6°C and a humidity of 8-10°C until cured; more preferably drying at a temperature of 2-6°C and a humidity of 8-10°C for 10-12 hours.

[0046] The operation described in step S6B to fill the microneedle anion membrane with pressure is preferably a vacuum treatment; more preferably, it is to pressurize in an environment of -80 to -100 for 15 to 20 minutes.

[0047] The drying conditions described in step S6B are preferably drying at a temperature of 2-6°C and a humidity of 8-10°C until cured; more preferably drying at a temperature of 2-6°C and a humidity of 8-10°C for 10-12 hours.

[0048] The drying conditions described in step S6C are preferably a temperature of 0–6°C and a humidity of 8–10°C; more preferably a temperature of 2–6°C and a humidity of 8–10°C.

[0049] The drying time in step S6C is preferably 10 to 12 hours.

[0050] The water used in this invention is preferably deionized water or purified water.

[0051] A compositional microneedle for treating gout is obtained by the above preparation method; it includes a needle body and a backing layer, the needle body is composed of an interconnected sustained-release layer and a burst-release layer, the sustained-release layer, the burst-release layer and the backing layer are connected in sequence; its needle length is 650-800 μm and the microneedle spacing is 250-350 μm.

[0052] The above-mentioned composition for treating gout is used in the preparation of drugs for treating gout.

[0053] Compared with existing technologies, the microneedles prepared by this invention address gout problems from multiple dimensions, and have the following advantages and beneficial effects:

[0054] (1) Reduce serum uric acid levels: By loading PEG-modified recombinant uricase and luteolin into the microneedle, the two can dissolve the crystalline crystals and promote their excretion; combining animal and plant sources, the two work synergistically to improve the stability and utilization of the recombinant uricase and the water solubility of luteolin, thus broadening the application areas of both.

[0055] (2) Inhibit the production of uric acid: The luteolin loaded in the microneedle body can inhibit the activity of xanthine oxidase, thereby inhibiting the production of uric acid;

[0056] (3) Anti-inflammatory and analgesic: By loading natural plant-derived active substances luteolin and dextrorotatory borneol into the microneedle body, the side effects of both are small when used;

[0057] (4) Enhanced Permeation and Highly Efficient Absorption: By combining the function of microneedles in penetrating the stratum corneum with the permeation-enhancing effect of dextrorotatory borneol, the prepared microneedle drug delivery achieves rapid "point-to-surface" delivery, accelerating transdermal drug absorption. First, when microneedles are used for transdermal drug delivery, self-administration and targeted, highly efficient delivery are possible, and it is painless. Second, dextrorotatory borneol disrupts the arrangement of keratinocytes, destroys the lipid bilayer structure of the stratum corneum, weakens the barrier function, and thus promotes the transdermal absorption of active substances. Preparing dextrorotatory borneol into a microemulsion and mixing it with active substances for treating gout can promote its diffusion into cells and improve its utilization rate. Attached Figure Description

[0058] Figure 1 This is a schematic diagram of the structure of a microneedle composition for treating gout according to the present invention; wherein, 1-sustained release layer, 2-burst release layer, 3-backing layer. Detailed Implementation

[0059] The present invention will be further described in detail below with reference to the embodiments and accompanying drawings, but the embodiments of the present invention are not limited thereto.

[0060] Unless otherwise specified, all reagents used in the examples are commercially available.

[0061] Example 1

[0062] (1) Preparation of PEG-modified recombinant uricase: Recombinant uricase protein (Chimeric mutant 7 protein) was obtained according to Example 3 of CN202211051614.3. 4.8 μg / μL of recombinant uricase protein was mixed with 5 kDa-mPEG-SPA at a mass ratio of 1:35 in a PBS (0.01 mol / L, pH 7.4) reaction system. Optimal modification was achieved by modification at 4°C for 6 h. After modification, the modified mixture was continuously ultrafiltered through a 10 kDa ultrafiltration tube to remove the hydrolysis product N-hydroxy-succinimide and excess free 5 kDa-PEG-SPA. The purity of the ultrafiltered product was confirmed to be greater than 90% by barium iodide staining. The yield of the PEG-modified recombinant uricase prepared by this method was 50-60%, and the concentration of the uricase was 40-50 mg / mL.

[0063] (2) Preparation of dextrorotatory borneol microemulsion: In a beaker, oil phase (caprylic / capric acid glycerol), surfactant (Tween 20), and co-surfactant (1,2-propanediol) were added in a mass ratio of 2:9:9. Then dextrorotatory borneol was added. The mixture was first sonicated at 35°C (250W, 40KHz) for 30 minutes to dissolve the dextrorotatory borneol. Then it was stirred on a magnetic stirrer (250r / min) for 5 minutes, allowed to stand for 10 minutes, and an appropriate amount of water was added. The mixture was stirred evenly to prepare the dextrorotatory borneol microemulsion. The contents of each component were as follows: dextrorotatory borneol 3%, caprylic / capric acid glycerol 4%, Tween 20 18%, 1,2-propanediol 18%, and the remainder was water.

[0064] (3) Preparation of luteolin-dextrin microemulsion solution: Add luteolin to the dextrin microemulsion obtained in step (2) at a mass ratio of 1:1 with dextrin, stir for 5 min on a magnetic stirrer (250 r / min), and let stand for 10 min to obtain luteolin-dextrin microemulsion solution.

[0065] (4) The microneedle patch composition comprises the following components by weight percentage:

[0066] The sustained-release layer contains 20% HAMA (methacrylamide hyaluronic acid), 5% PEG-modified recombinant uricase preparation, 10% luteolin-dextrin microemulsion solution, and the remainder is deionized water.

[0067] Burst-release layer solution: PVP K90 20%.

[0068] Backing layer solution: PVP K90 20%, PVA 1788 10%, glycerol 1.5%, balance deionized water.

[0069] (5) The specific preparation method of the microneedle patch of the composition is as follows (% are all mass percentages):

[0070] 1) Preparation of needle excipient solution: Mix HAMA with deionized water in proportion, stir in a water bath at 75°C for 6 hours until the solution is clear and transparent to obtain the needle excipient solution, which is the matrix solution.

[0071] 2) Preparation of sustained-release layer solution: The PEG-modified recombinant uricase preparation and luteolin-dextrin microemulsion solution were added to the matrix solution while stirring to obtain the sustained-release layer solution; the contents of each component are as follows: PEG-modified recombinant uricase preparation 5%, luteolin-dextrin microemulsion solution 10%, and HAMA concentration 20%.

[0072] 3) Preparation of burst layer solution: PVP K90 and deionized water are mixed in proportion and stirred in a water bath at 75°C for 6 hours until the solution is clear and transparent to obtain the burst layer solution; wherein, the concentration of PVP K90 in the burst layer solution is 20%.

[0073] 4) Preparation of backing layer solution: PVP K90, PVA 1788 and glycerol are mixed with deionized water and stirred in a water bath at 75°C for 6 hours until the solution is clear and transparent to obtain the backing layer solution; the contents of each substance are as follows: PVP K90 20%, PVA1788 10%, glycerol 1.5%, and the balance is deionized water.

[0074] 5) Add the sustained-release layer solution obtained in step 2) into the microneedle anion membrane, and treat it under vacuum of -0.9 (100 x Kpa) (equivalent to -90 Kpa) for 15 min. Repeat the treatment twice to fill the microneedle anion membrane. Scrape off the excess liquid, and then dry it at a temperature of 2℃ and a humidity of 10° for 12 h to solidify and form a microneedle sustained-release layer.

[0075] 6) Add the burst-release layer solution obtained in step 3) onto the microneedle sustained-release layer, and treat it under vacuum at -1 (100 x Kpa) for 15 min, repeating the treatment twice to fill the microneedle anion membrane. Scrape off the excess liquid, and then dry it at 2°C and 10°C for 12 h to solidify and form the microneedle burst-release layer.

[0076] 7) Spread the backing layer solution obtained in step 4) on the microneedle release layer, and dry the entire microneedle at a temperature of 2°C and a humidity of 10°C for 12 hours to solidify and form the composite microneedle.

[0077] 8) Remove the microneedle patch from the mold to obtain the composite microneedles, such as... Figure 1 As shown, it includes a needle body and a backing layer 3. The needle body consists of an interconnected sustained-release layer 1 and a burst-release layer 2. The microneedles are conical in shape, with a needle length of 650 μm and a microneedle spacing of 250 μm.

[0078] Example 2

[0079] (1) Preparation of PEG-modified recombinant uricase preparation: Same as in Example 1.

[0080] (2) Preparation of dextrorotatory borneol microemulsion: In a beaker, oil phase (caprylic / capric acid glycerol), surfactant (Tween 20), and co-surfactant (1,2-propanediol) were added in a mass ratio of 2:6:2.5. Then dextrorotatory borneol was added. The mixture was first sonicated at 35°C (250W, 40KHz) for 30 minutes to dissolve the dextrorotatory borneol. Then it was stirred on a magnetic stirrer (200r / min) for 5 minutes, allowed to stand for 10 minutes, and an appropriate amount of water was added. The microemulsion was prepared by stirring evenly. The contents of each component were as follows: dextrorotatory borneol 3%, caprylic / capric acid glycerol glycerol 8%, Tween 20 24%, 1,2-propanediol 10%, and the remainder was water.

[0081] (3) Preparation of luteolin-dextrin microemulsion solution: Add luteolin to the dextrin microemulsion obtained in step (2) at a mass ratio of 1:1 with dextrin, stir for 5 min on a magnetic stirrer (250 r / min), and let stand for 10 min to obtain luteolin-dextrin microemulsion solution.

[0082] (4) The microneedle patch composition comprises the following components by weight percentage:

[0083] The sustained-release layer contains 25% HAMA (methacrylamide hyaluronic acid), 7% PEG-modified recombinant uricase preparation, 15% luteolin-dextrin microemulsion solution, and the remainder is deionized water.

[0084] Burst-release layer solution: PVP K30 25%.

[0085] Backing solution: PVP K90 20%, chitosan 10%, glycerol 1.5%, balance deionized water.

[0086] (5) The specific preparation method of the microneedle patch of the composition is as follows (% are all mass percentages):

[0087] 1) Preparation of needle excipient solution: Mix HAMA with deionized water in proportion, stir in a water bath at 75°C for 6 hours until the solution is clear and transparent to obtain the needle excipient solution, which is the matrix solution;

[0088] 2) Preparation of sustained-release layer solution: The PEG-modified recombinant uricase preparation and luteolin-dextrin microemulsion solution were added to the matrix solution while stirring to obtain the sustained-release layer solution; the contents of each component are as follows: PEG-modified recombinant uricase preparation 7%, luteolin-dextrin microemulsion solution 15%, and HAMA concentration 25%.

[0089] 3) Preparation of burst layer solution: PVP K30 and deionized water are mixed in proportion and stirred in a water bath at 75°C for 6 hours until the solution is clear and transparent to obtain the burst layer solution; wherein, the concentration of PVP K30 in the burst layer solution is 25%.

[0090] 4) Preparation of backing layer solution: PVP K90, chitosan, glycerol and part of deionized water are mixed in proportion and stirred in a water bath at 75°C for 6 hours until the solution is clear and transparent to obtain the backing layer solution; wherein, the content of each substance is as follows: PVP K90 20%, chitosan 10%, glycerol 1.5%, and the balance is deionized water.

[0091] 5) Add the sustained-release layer solution obtained in step 2) into the microneedle anion membrane, and treat it under vacuum of -0.9 (100 x Kpa) for 20 min. Repeat the treatment twice to fill the microneedle anion membrane. Scrape off the excess liquid, and then dry it at a temperature of 4°C and a humidity of 9°C for 11 h to solidify and form a microneedle sustained-release layer.

[0092] 6) Add the burst-release layer solution obtained in step 3) onto the microneedle sustained-release layer, and treat it under vacuum of -0.9 (100 x Kpa) for 20 min, repeating the treatment twice to fill the microneedle anion membrane. Scrape off the excess liquid, and then dry it at a temperature of 4°C and a humidity of 9°C for 11 h to solidify and form the microneedle burst-release layer.

[0093] 7) Spread the backing layer solution obtained in step 4) on the microneedle release layer, and dry the entire microneedle at a temperature of 4°C and a humidity of 9°C for 12 hours to solidify and form the composite microneedle.

[0094] 8) Remove the microneedle patch from the mold to obtain the composite microneedles. The microneedles are conical in shape, 750 μm in length, and 300 μm in spacing.

[0095] Example 3

[0096] (1) Preparation of PEG-modified recombinant uricase preparation: Same as in Example 1.

[0097] (2) Preparation of dextrorotatory borneol microemulsion: In a beaker, oil phase (caprylic / capric acid glycerol), surfactant (Tween 40), and co-surfactant (1,2-propanediol) were added in a mass ratio of 2:5.5:3. Then dextrorotatory borneol was added. The mixture was first sonicated at 35°C (250W, 40KHz) for 30 minutes to dissolve the dextrorotatory borneol. Then it was stirred on a magnetic stirrer (200r / min) for 5 minutes, allowed to stand for 10 minutes, and an appropriate amount of water was added. The microemulsion was prepared by stirring evenly. The contents of each component were as follows: dextrorotatory borneol 3.5%, caprylic / capric acid glycerol glycerol 8%, Tween 40 22%, 1,2-propanediol 12%, and the remainder was water.

[0098] (3) Preparation of luteolin-dextrin microemulsion solution: Add luteolin to the dextrin microemulsion obtained in step (2) at a mass ratio of 1:1 with dextrin, stir for 5 min on a magnetic stirrer (250 r / min), and let stand for 10 min to obtain luteolin-dextrin microemulsion solution.

[0099] (4) The microneedle patch composition comprises the following components by weight percentage:

[0100] The sustained-release layer contains 30% HAMA (methacrylamide hyaluronic acid), 10% PEG-modified recombinant uricase preparation, 12% luteolin-dextrin microemulsion solution, and the remainder is deionized water.

[0101] Burst-release layer solution: PVA1788 30%.

[0102] Backing layer solution: PVP K90 20%, PVA 1788 10%, glycerol 1.5%, balance deionized water.

[0103] (5) The specific preparation method of the microneedle patch of the composition is as follows (% are all mass percentages):

[0104] 1) Preparation of needle excipient solution: Mix HAMA with deionized water in proportion, stir in a water bath at 75°C for 6 hours until the solution is clear and transparent to obtain the needle excipient solution, which is the matrix solution;

[0105] 2) Preparation of sustained-release layer solution: The PEG-modified recombinant uricase preparation and luteolin-dextrin microemulsion solution were added to the matrix solution while stirring to obtain the sustained-release layer solution; the contents of each component are as follows: PEG-modified recombinant uricase preparation 10%, luteolin-dextrin microemulsion solution 12%, and HAMA concentration 30%.

[0106] 3) Preparation of burst layer solution: PVA 1788 is mixed with deionized water in proportion, and stirred in a water bath at 75°C for 6 hours until the solution is clear and transparent to obtain the burst layer solution; wherein, the concentration of PVA 1788 in the burst layer solution is 30%.

[0107] 4) Preparation of backing layer solution: Mix PVP K90, PVA 1788, glycerol and deionized water, stir in a water bath at 75°C for 6 hours until the solution is clear and transparent to obtain the backing layer solution; wherein, the content of each substance is as follows: PVP K90 20%, PVA 1788 10%, glycerol 1.5%, and the balance is deionized water.

[0108] 5) Add the sustained-release layer solution obtained in step 2) into the microneedle anion membrane, and treat it under vacuum of -0.8 (100 x Kpa) for 18 min, repeat the treatment twice to fill the microneedle anion membrane, scrape off the excess liquid, and then dry it at a temperature of 6°C and a humidity of 8°C for 10 h to solidify and form a microneedle sustained-release layer.

[0109] 6) Add the burst-release layer solution obtained in step 3) onto the microneedle sustained-release layer, and treat it under vacuum of -0.8 (100 x Kpa) for 15 min, repeating the treatment twice to fill the microneedle anion membrane. Scrape off the excess liquid, and then dry it at a temperature of 6°C and a humidity of 8°C for 11 h to solidify and form the microneedle burst-release layer.

[0110] 7) Spread the backing layer solution obtained in step 4) on the microneedle burst release layer, and place the entire microneedle at a temperature of 6°C and a humidity of 8°C for 12 hours to dry and solidify to form the composite microneedle.

[0111] 8) Remove the microneedle patch from the mold to obtain the composite microneedles. The microneedles are conical in shape, 800 μm in length, and 350 μm in spacing.

[0112] Comparative Example 1

[0113] Microneedles of the composition were prepared according to the same preparation method as in Example 1, except that the recombinant uricase preparation without PEG modification was used (the same amount of water was used to replace the recombinant uricase preparation with PEG modification). The sustained-release layer solution was added to the microneedle mold according to step (2) to obtain Comparative Example 1.

[0114] Comparative Example 2

[0115] Microneedles of the composition were prepared according to the same preparation method as in Example 1, except that the recombinant uricase preparation and dextrorotatory borneol microemulsion without PEG modification were not used (the same amount of water was used to replace the recombinant uricase preparation and dextrorotatory borneol microemulsion with PEG modification). The sustained-release layer solution was added to the microneedle mold according to step (2) to prepare Comparative Example 2.

[0116] Comparative Example 3

[0117] Microneedles of the composition were prepared according to the same preparation method as in Example 2, except that they did not contain luteolin (the same amount of water was used instead of luteolin solution). The sustained-release layer solution was added to the microneedle mold according to step (2) to prepare Comparative Example 3.

[0118] Comparative Example 4

[0119] Microneedles of the composition were prepared according to the same preparation method as in Example 2, except that they did not contain luteolin and dextrorotatory borneol microemulsion (the same amount of water was used to replace the luteolin solution and dextrorotatory borneol microemulsion). The sustained-release layer solution was added to the microneedle mold according to step (2) to prepare Comparative Example 4.

[0120] Comparative Example 5

[0121] The composition microneedles were prepared according to the same active ingredient for treating gout and the same preparation method as in Example 3, except that they did not contain dextrorotatory borneol microemulsion (the same amount of water was used instead of dextrorotatory borneol microemulsion). The sustained-release layer solution was added to the microneedle mold according to step (2) to prepare Comparative Example 5.

[0122] Comparative Example 6

[0123] The composition microneedles were prepared according to the same active ingredient for treating gout and the same preparation method as in Example 3, except that the recombinant uricase preparation without PEG modification was not used (the same amount of water was used instead of the recombinant uricase preparation with PEG modification). The sustained-release layer solution was added to the microneedle mold according to step 2) to prepare the microneedle, thus obtaining Comparative Example 6.

[0124] Comparative Example 7

[0125] Microneedles were prepared using the same active ingredient for treating gout and the same preparation method as in Example 3, except that they did not contain luteolin (the same amount of water was used instead of luteolin solution). The sustained-release layer solution was added to the microneedle mold in step 2) to prepare the microneedle, thus obtaining Comparative Example 7.

[0126] Comparative Example 8

[0127] Microneedles of the composition were prepared according to the same preparation method as in Example 3, except that the recombinant uricase preparation and luteolin were not modified with PEG (the same amount of water was used to replace the recombinant uricase preparation and luteolin modified with PEG). The sustained-release layer solution was added to the microneedle mold according to step (2) to prepare Comparative Example 8.

[0128] Example Effect Test

[0129] The application effects of the above three embodiments were tested, and the specific testing methods are as follows:

[0130] Ninety volunteers (aged 25-65 years, 45 males and 45 females) were selected. The causes of gout attacks included acute arthritis (40 cases), tophi (25 cases), tophi-related chronic arthritis (20 cases), and other causes (5 cases). The volunteers were randomly divided into 9 groups (Groups 1, 2, and 3 used Example 1, Comparative Example 1, and Comparative Example 2, respectively; Groups 4, 5, and 6 used Example 2, Comparative Example 3, and Comparative Example 4, respectively; Groups 7, 8, and 9 used Example 3, Comparative Example 5, and Comparative Example 6, respectively), with 10 volunteers in each group. The above-described composition was applied to the painful areas of the volunteers' skin and joints using microneedles. Examples 1-3 and Comparative Examples 1-6 were each applied for 48 hours each time. This was continued for one month, twice a week. The criteria for evaluating the treatment effect were as follows: (1) Cured: no pain, swelling, redness or inflammation at the joint site, and the pain disappeared; (2) Significantly effective: no pain, swelling, redness or inflammation at the joint site, and mild pain lasting <2 hours; (3) Effective: significant improvement in pain, swelling, redness or inflammation at the joint site, and mild pain lasting <8 hours; (4) Ineffective: no significant change in pain, swelling, redness or inflammation at the joint site, and persistent pain lasting >8 hours. The results are shown in Table 1.

[0131] Table 1 Comparison of the effects of gout treatment [n(x%)]

[0132]

[0133]

[0134] Note: n represents the number of people, and x% represents the percentage.

[0135] As shown in Table 1, compared with Comparative Examples 1-4, Examples 1-2 of the present invention significantly reduced symptoms such as pain, swelling, redness, or inflammation at the joint site, and the pain disappeared. The effective rate of gout treatment was: Example 1 > Comparative Example 1 > Comparative Example 2, Example 2 > Comparative Example 3 > Comparative Example 4, indicating that the PEG-modified recombinant uricase preparation and luteolin alone can reduce uric acid accumulation and promote uric acid excretion; Example 3 > Comparative Example 5 and Comparative Example 8 had an effective rate of 30%, indicating that dextrorotatory borneol has a permeation-enhancing effect, promoting transdermal absorption of the drug. Simultaneously, dextrorotatory borneol, as an analgesic component, has a strong inhibitory effect on the pain and redness caused by gout; Comparative Example 5 > Comparative Examples 6 and 7, indicating that the PEG-modified recombinant uricase preparation and luteolin have a synergistic effect, and the combined microneedle patch treatment has significant therapeutic effects.

[0136] Meanwhile, Examples 1-3 of the present invention can significantly shorten the treatment cycle. After one week of continuous treatment, the average cure rate of 46.6% is significantly higher than that of the control group of 12.5%. After one week of treatment, no adverse symptoms such as allergies or discomfort occurred.

[0137] Working Principle: This invention addresses gout from multiple dimensions by incorporating PEG-modified recombinant uricase, luteolin, and dextrorotatory borneol microemulsion into the sustained-release layer of microneedles. This approach reduces serum uric acid levels, inhibits uric acid production, reduces inflammation and pain, and promotes efficient absorption. Simultaneously, the PEG-modified recombinant uricase and luteolin synergistically enhance the therapeutic effect on gout. Furthermore, the microneedles are prepared using an active separation cryo-treatment process. After the microneedles are inserted into the skin, the burst-release layer dissolves rapidly, while the sustained-release layer remains within the skin, allowing for continuous drug release.

[0138] The above embodiments are preferred embodiments of the present invention. However, those skilled in the art should understand that changes can be made to the embodiments without departing from the spirit of the present invention. Of course, this should not be construed as limiting the scope of the present invention. Therefore, any modifications, substitutions, combinations, or simplifications made that deviate from the spirit and principle of the present invention should be considered equivalent substitutions and still fall within the scope of the present invention.

Claims

1. A method for preparing a compositional microneedle for treating gout, characterized in that... Includes the following steps: S1. Preparation of dextrorotatory borneol microemulsion: The oil phase, surfactant, co-surfactant and dextrorotatory borneol are mixed and mixed evenly to obtain solution A; water droplets are added to the solution while stirring, and stirring is continued after the water droplets are added. After standing, dextrorotatory borneol microemulsion is obtained; wherein, the content of each component by mass percentage is as follows: dextrorotatory borneol 3-3.5%, oil phase 4-8%, surfactant 18-22%, co-surfactant 10-18%, water balance; S2. Preparation of luteolin-dextrin microemulsion solution: Luteolin and the dextrin microemulsion obtained in step S1 are mixed evenly and allowed to stand to obtain luteolin-dextrin microemulsion solution; wherein, the content of luteolin in the luteolin-dextrin microemulsion solution is 3-3.5% by mass. S3. Preparation of sustained-release layer solution: Mix uricase, the luteolin-dextrin microemulsion solution prepared in step S2, and the aqueous solution of needle excipient to obtain a sustained-release layer solution; wherein, the amounts of each component are as follows by mass percentage: uricase 5-10%, luteolin-dextrin microemulsion solution 10-15%, needle excipient 20-30%, and water as the balance; S4. Preparation of burst-release layer solution: Mix the soluble polymer with water until homogeneous to obtain a burst-release layer solution; wherein the concentration of the soluble polymer in the burst-release layer solution is 20-30% by mass. S5. Preparation of backing layer solution: Mix soluble polymer, plasticizer and water evenly to obtain backing layer solution; wherein, the content of each component by mass percentage is as follows: soluble polymer 25-35%, plasticizer 1-2%, water is the balance; S6. Preparation of microneedles: A. Add the sustained-release layer solution prepared in step S3 into the microneedle anion membrane to fill the microneedle anion membrane, scrape off the excess liquid, and dry and solidify to form a microneedle sustained-release layer; B. Add the burst-release layer solution prepared in step S4 to the microneedle sustained-release layer in step A, so that it fills the microneedle anion membrane, scrape off the excess liquid, and dry and solidify to form the microneedle burst-release layer. C. Spread the backing layer solution prepared in step S5 onto the microneedle release layer obtained in step B, and dry the entire microneedle to obtain the composition microneedle for treating gout.

2. The method for preparing the composition microneedles for treating gout according to claim 1, characterized in that: The oil phase mentioned in step S1 is polyethylene glycol glycerol octanoate-capric acid; The surfactant mentioned in step S1 is at least one of Tween 20 and Tween 40; The co-surfactant mentioned in step S1 is at least one of anhydrous ethanol and 1,2-propanediol.

3. The method for preparing the compositional microneedles for treating gout according to claim 2, characterized in that: The oil phase, surfactant, and co-surfactant mentioned in step S1 are in a mass ratio of 2: 5.5~9 : 2.5~9 ratio.

4. The method for preparing the compositional microneedles for treating gout according to claim 1, characterized in that: In step S2, luteolin and dextrorotatory borneol are mixed in a mass ratio of 1:1 in the luteolin-dextrin microemulsion solution.

5. The method for preparing the compositional microneedles for treating gout according to claim 1, characterized in that: The uricase mentioned in step S3 is a PEG-modified uricase; The needle-forming material mentioned in step S3 is methacrylamide hyaluronic acid; The soluble polymer mentioned in step S4 is at least one of polyvinylpyrrolidone and polyvinyl alcohol; The soluble polymer mentioned in step S5 is at least one of polyvinylpyrrolidone, polyvinyl alcohol, and chitosan; The plasticizer mentioned in step S5 is glycerin.

6. The method for preparing the compositional microneedles for treating gout according to claim 5, characterized in that: The uricase described in step S3 is a humanized human-porcine chimeric uricase mutant; The aqueous solution of the needle-shaped material mentioned in step S3 is prepared by the following steps: under heating conditions, the needle-shaped material is dissolved in water to obtain the aqueous solution of the needle-shaped material; The polyvinylpyrrolidone is at least one of polyvinylpyrrolidone K30 and polyvinylpyrrolidone K90; The polyvinyl alcohol mentioned is polyvinyl alcohol 1788; The soluble polymer mentioned in step S5 is a composition formed by polyvinyl alcohol and chitosan with polyvinylpyrrolidone.

7. The method for preparing the compositional microneedles for treating gout according to claim 1, characterized in that: The mixing method described in step S1 is ultrasound; The stirring speed mentioned in step S1 is 200-300 rpm; The stirring time mentioned in step S1 is 3 to 8 minutes; The settling time mentioned in step S1 is 8 to 15 minutes; The mixing process described in step S2 is as follows: stir at 200-300 rpm for 3-8 minutes, and then let stand for 8-15 minutes; The mixing conditions described in step S4 are stirring in a water bath at 70–80°C; The mixing conditions described in step S5 are stirring in a water bath at 70-80°C; The operation described in step S6A to fill the microneedle anion membrane is a pressurized vacuum treatment; The drying conditions described in step S6A are drying at a temperature of 2-6°C and a humidity of 8-10°C until cured; The operation described in step S6B to fill the microneedle anion membrane is a pressurized vacuum treatment; The drying conditions described in step S6B are drying at a temperature of 2-6°C and a humidity of 8-10°C until cured; The drying conditions described in step S6C are a temperature of 0–6°C and a humidity of 8–10°C.

8. The method for preparing the compositional microneedles for treating gout according to claim 7, characterized in that: The pressure vacuum treatment conditions are as follows: pressurize in an environment of -80 to -100 MPa for 15 to 20 minutes; The drying conditions described in step S6A are: drying at a temperature of 2-6°C and a humidity of 8-10°C for 10-12 hours; The drying conditions described in step S6B are: drying at a temperature of 2-6°C and a humidity of 8-10°C for 10-12 hours.

9. A compositional microneedle for treating gout, characterized in that: It is obtained by the preparation method according to any one of claims 1 to 8; it includes a needle body and a backing layer, the needle body being composed of an interconnected sustained-release layer and a burst-release layer.

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