Polypeptide drug-loaded microneedle patch for treating rheumatoid arthritis

Abstract

The invention discloses a micro-needle transdermal drug delivery patch for treating rheumatoid arthritis and a preparation method of the micro-needle transdermal drug delivery patch. An active drug component melittin is contained in a micro-needle. The polymer microneedle disclosed by the invention can effectively puncture the skin cuticle to enter the epidermal layer, and the medicine stored in the microneedle is delivered into the skin tissue, so that the transdermal delivery efficiency of the medicine is greatly improved. Animal experiments show that the microneedle has enough mechanical strength to puncture the skin, so that the pathological manifestation of a rheumatoid arthritis animal model is effectively inhibited, and a good treatment effect is shown. The preparation method of the microneedle is simple and convenient, and has a good clinical transformation prospect.

Description

Technical Field

[0001] The invention relates to the technical field of microneedle patch preparations for transdermal administration, in particular to a polymer microneedle loaded with melittin and application thereof in rheumatoid arthritis, belonging to the technical field of rheumatoid arthritis treatment. Background Art

[0002] Rheumatoid arthritis (RA) is an autoimmune disease characterized by chronic synovial inflammation. The worldwide prevalence is about 1%. This disease can cause joint swelling, deformity and even disability, greatly affecting the quality of life of patients. Currently, RA is usually treated with oral or injected nonsteroidal anti-inflammatory drugs (NSAIDs) or corticosteroids. These strategies can reduce inflammation and delay the progression of RA by interfering with inflammation-related pathways, such as reducing the levels of proinflammatory cytokines. However, patient compliance with these traditional strategies is low due to pain or infection. In addition, injections or frequent oral administration can cause serious gastrointestinal side effects, and the harsh gastrointestinal environment and first-pass effect of the liver can easily lead to significantly low drug bioavailability.

[0003] Microneedles are micro-needle structures with a length of 100-1000μm made by micro-machining. When the needle body punctures the skin stratum corneum barrier, it will not touch the nerve tissue and blood vessels. Therefore, microneedle delivery of drugs is almost painless and minimally invasive, and patients have good compliance. Microneedle drug delivery has many advantages, such as painless drug delivery, which can greatly increase the compliance of users; microneedle drug delivery has extremely low professional and technical requirements, and patients can administer drugs themselves; the drugs in the microneedles exist in solid form, have good stability, and are easy to transport and store. However, the existing polymer microneedles are mostly prepared by dissolving the drug and the matrix in a solvent, requiring the drug and the matrix to have the same solubility (soluble in water or soluble in organic solvents). When it is necessary to carry water-soluble and water-insoluble drugs at the same time, other complex methods need to be adopted, such as preparing nanoparticles, liposomes, inclusion compounds, etc. These methods not only increase the difficulty of preparing microneedles, but also further reduce the drug loading, and the mechanical strength of the microneedles is also easily affected.

[0004] Previous studies, including several clinical trials conducted in East Asia, have shown that bee venom acupuncture has a good therapeutic effect on arthritis diseases with fewer side effects. Further studies have shown that the water-soluble components of bee venom can play an anti-inflammatory role in RA and delay disease progression. A 26-amino acid polypeptide fragment in the bee venom component is the main component of the immunomodulatory and anti-RA effects of bee venom. However, intravenous injection of bee venom or injection of purified melittin has the risk of severe hemolysis, and these shortcomings seriously limit the application of melittin as an anti-rheumatoid arthritis drug. Transdermal delivery of melittin, such as using microneedles, may overcome the above shortcomings. So far, there has been no report of using microneedle delivery of melittin to treat rheumatoid arthritis. Summary of the invention

[0005] In order to solve the above problems, the present invention prepares a polymer microneedle loaded with bee venom peptide and studies its therapeutic effect on rheumatoid arthritis. On the one hand, the present invention avoids the risk of hemolysis during the use of bee venom peptide by loading bee venom peptide into microneedles for transdermal delivery; on the other hand, the use of methacrylic-modified hyaluronic acid to prepare microneedles can give the drug controlled release properties, significantly improving the therapeutic effect of the drug. At the same time, the use of microneedles for transdermal administration avoids the toxic and side effects of traditional oral treatments and the pain associated with injection administration and the hidden dangers of low patient compliance.

[0006] The microneedle patch of the present invention comprises a backing layer and a needle body, wherein the needle body comprises a needle-forming matrix material and a drug, wherein the needle-forming matrix material comprises a high molecular polymer, and the drug comprises an anti-rheumatoid arthritis drug melittin.

[0007] In order to achieve the above object, the present invention adopts the following technical solutions:

[0008] 1) dissolving the biocompatible polymer matrix material in water and completely dissolving it by stirring;

[0009] 2) adding the above suspension into a microneedle mold, using pressurized gas to press the solution into the microneedle mold, placing it in a drying kettle containing anhydrous silica gel and drying it at room temperature for 2 hours to form a microneedle tip;

[0010] 3) Adding a biocompatible polymer matrix solution into the mold and drying for 2 hours to form a microneedle base;

[0011] 4) The microneedles were placed under 365 nm ultraviolet light for cross-linking for 15 seconds, and then finally peeled off to obtain a microneedle patch.

[0012] The microneedle patch for treating rheumatoid arthritis of the present invention, the biocompatible polymer matrix material includes hyaluronic acid, polyvinyl pyrrolidone, collagen, silk protein, gelatin, hydroxypropyl methylcellulose, chondroitin sulfate, dextrin, polyvinyl alcohol, carboxymethyl cellulose, carboxymethyl chitosan, dextran sulfate, glycogen, amylose, dextran, hydroxypropyl cellulose, glycoside, chitosan, polyester, PHA, PHBV, PHP, PHH, PHA-PEG, poly 4-hydroxy acid, poly α-hydroxy acid, poly β-hydroxy acid, At least one of poly 4-hydroxybutyrate, poly 4-hydroxyvalerate, poly 4-hydroxycaproate, polyester amide, polycaprolactone, polylactide, polyglycolide, PLGA, polydioxanone, polyorthoester, polyetherester, polyanhydride, glycolic acid-trimethylene carbonate copolymer, polyphosphate, polyphosphate urethane, polyamino acid, polycyanoacrylate, polytrimethylene carbonate, polyimino carbonate, polytyrosine carbonate, polycarbonate, polytyrosine aryl ester, polyalkylene oxalate, polyphosphocreatine and derivatives of the above materials.

[0013] In some embodiments, the needle-forming matrix material is preferably one of hyaluronic acid, polyvinyl pyrrolidone, and silk fibroin.

[0014] Further preferably, the needle-forming matrix material is preferably hyaluronic acid, and the molecular weight of hyaluronic acid is 10k-300k and the concentration is 20-50%.

[0015] The microneedle patch of the present invention is suitable for treating rheumatoid arthritis. The anti-rheumatoid arthritis polypeptide drug is melittin, and the concentration of melittin is 1%-10% (m / v).

[0016] Compared with the prior art solutions, the present invention has the following beneficial effects:

[0017] Rheumatoid arthritis is a chronic autoimmune disease that requires long-term and multiple medications, has the risk of repeated courses of disease, and is prone to relapse after discontinuation of medication. The microneedle patch prepared by the present invention uses methacrylic acid-modified hyaluronic acid to load bee venom peptide, and can show good sustained-release properties after transdermal delivery. In vivo experiments show that the sustained-release microneedle can be continuously released in the skin for 1 week after administration. The microneedle patch used in the present invention has a good therapeutic effect on mouse rheumatoid arthritis and has the prospect of new drug development.

[0018] The present invention utilizes a microneedle patch to treat rheumatoid arthritis patients who need long-term medication, thereby solving the problems of toxic and side effects of traditional treatment methods, pain associated with injection administration, and low patient compliance. BRIEF DESCRIPTION OF THE DRAWINGS

[0019] Figure 1This is the microneedle patch for treating rheumatoid arthritis prepared in Example 1 of the present invention.

[0020] Figure 2 This is the inflammation score of mice with rheumatoid arthritis treated with the microneedle patch according to Example 2 of the present invention. DETAILED DESCRIPTION

[0021] The following examples are further explanations of the present invention, and researchers or technicians in this field can understand the advantages of the present invention based on the content of the present invention. However, it should be understood by those skilled in the art that the protection scope of the present invention is not limited to the following specific embodiments. The terms used in the examples of the present invention are to describe specific embodiments, not to limit the protection scope of the present invention.

[0022] The present invention does not limit the sources of the raw materials used. Unless otherwise specified, the raw materials used in the present invention are common commercial products in the technical field.

[0023] Example 1

[0024] Melittin and methacrylic acid-modified hyaluronic acid (including 2-mercaptobenzoic acid and photoinitiator-2959) were dissolved in water;

[0025] The suspension was added into the microneedle mold, and the solution was pressed into the microneedle mold using pressurized gas, and then placed in a drying kettle containing anhydrous silica gel and dried at room temperature for 2 hours to form a microneedle tip;

[0026] Add hyaluronic acid solution to the microneedle mold and dry for 2 hours to form a base;

[0027] The microneedles were placed under 365nm ultraviolet light for cross-linking for 15s, and then finally peeled off to obtain a microneedle patch.

[0028] The prepared microneedles Figure 1 It can be seen that the prepared bee venom peptide-loaded polymer microneedles have regular shapes, intact needle tips, and a rupture pressure of 27 MPa.

[0029] Example 2

[0030] Establishment of AIA mouse model: SD rats were selected and 80 μl of complete Freund's adjuvant containing 10 mg / ml Mycobacterium tuberculosis was subcutaneously injected into the soles of their feet to complete the establishment of the AIA model.

[0031] Treatment plan: The mice were randomly divided into four groups, namely: control group (Control), subcutaneous injection group (SC), HA microneedle group loaded with melittin (Mel-HA-MN), and microneedle group loaded with melittin MeHA (Mel-MeHA-MN). The healthy group mice were not treated in any way, and the other three groups were CIA model mice. From the fourth day after modeling, each group received subcutaneous injection of melittin, HA microneedle loaded with melittin, and microneedle group loaded with melittin MeHA, respectively. The administration method was to use microneedles to administer the drug on the back of the mouse, and the drug was administered every other day for a total of 8 times.

[0032] Monitoring of therapeutic effects: Starting from day 0 after modeling, the plantar thickness of mice was measured with a vernier caliper every day, and the joint swelling of rats in each group was scored. Figure 2 The results showed that the plantar thickness and inflammatory index of the MeHA microneedle group loaded with bee venom peptide were significantly lower than those of the control group, subcutaneous injection group, and HA microneedle group loaded with bee venom peptide, indicating that sustained-release microneedles loaded with bee venom peptide can exert better therapeutic effects.

[0033] In summary, the present invention uses a polymer microneedle patch to deliver the anti-rheumatoid arthritis drug melittin to the skin, effectively preventing the swelling of the soles of the feet of rheumatoid arthritis model mice, significantly reducing the inflammation score of the model mice, and exerting the best therapeutic effect on the rheumatoid arthritis model mice. In addition, the present invention uses a microneedle patch to administer medication to a group of patients with rheumatoid arthritis who need long-term medication, solving the problems of toxic and side effects of traditional oral treatments and pain associated with injection administration and low patient compliance.

[0034] The above is only a preferred embodiment of the present invention, and does not limit the present invention in any form. Although the present invention has been disclosed as a preferred embodiment as above, it is not used to limit the present invention. Any technician familiar with this profession can make some changes or modify the technical contents disclosed above into equivalent embodiments of equivalent changes without departing from the scope of the technical solution of the present invention. However, any simple modification, equivalent change and modification made to the above embodiments according to the technical essence of the present invention without departing from the content of the technical solution of the present invention still fall within the scope of the technical solution of the present invention.

Claims

1. A microneedle transdermal drug delivery patch for treating rheumatoid arthritis, characterized in that: The microneedle patch comprises a backing layer and a needle body, the needle body comprises a needle-forming matrix material and a drug, the needle-forming matrix material comprises a biocompatible polymer matrix material, and the drug comprises an anti-rheumatoid arthritis polypeptide drug.

2. The microneedle for treating rheumatoid arthritis according to claim 1, characterized in that The biocompatible polymer matrix material includes hyaluronic acid, polyvinyl pyrrolidone, collagen, silk protein, gelatin, hydroxypropyl methylcellulose, chondroitin sulfate, dextrin, polyvinyl alcohol, carboxymethyl cellulose, carboxymethyl chitosan, dextran sulfate, glycogen, amylose, dextran, hydroxypropyl cellulose, glycoside, chitosan, polyester, PHA, PHBV, PHP, PHH, PHA-PEG, poly 4-hydroxy acid, poly α-hydroxy acid, poly β-hydroxy acid, poly 4-hydroxybutyrate, poly 4 -hydroxyvalerate, poly 4-hydroxycaproate, polyester amide, polycaprolactone, polylactide, polyglycolide, PLGA, polydioxanone, polyorthoester, polyetherester, polyanhydride, glycolic acid-trimethylene carbonate copolymer, polyphosphate, polyphosphate urethane, polyamino acid, polycyanoacrylate, polytrimethylene carbonate, polyimino carbonate, polytyrosine carbonate, polycarbonate, polytyrosine aryl ester, polyalkylene oxalate, polyphosphocreatine and at least one of the derivatives of the above materials.

3. The microneedle for treating rheumatoid arthritis according to claim 1, characterized in that: The anti-rheumatoid arthritis polypeptide drug is melittin.

4. A method for preparing a microneedle for treating rheumatoid arthritis according to any one of claims 1 to 4, characterized in that: The method for preparing the microneedle comprises the following steps: 1) dissolving bee venom peptide and a biocompatible polymer matrix material in water, and completely dissolving them by stirring; 2) adding the above aqueous solution into a microneedle mold, using pressurized gas to press the solution into the microneedle mold, placing it in a drying kettle containing anhydrous silica gel and drying it at room temperature for 2 hours to form a microneedle tip; 3) Adding a biocompatible polymer matrix solution into the mold and drying for 2 hours to form a microneedle base; 4) The microneedles were placed under 365 nm ultraviolet light for cross-linking for 15 seconds, and then finally peeled off to obtain a microneedle patch.

5. The method for preparing the microneedle according to claim 5, characterized in that: The concentration of the anti-rheumatoid arthritis polypeptide drug melittin is 1%-10% (m / v).

6. Use of the microneedle transdermal drug delivery patch according to claim 1 in the preparation of anti-rheumatoid arthritis drugs.

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