A lappaconitine A microneedle composition, a microneedle prepared therefrom, a preparation method and application

Abstract

The application discloses a lappaconitine microneedle composition, a microneedle prepared from the lappaconitine microneedle composition, a preparation method and application of the lappaconitine microneedle composition. The lappaconitine microneedle composition contains 10-40% of lappaconitine inclusion compound and 60-90% of biocompatible polymer material in terms of mass percentage. The lappaconitine inclusion compound is selected from lappaconitine inclusion compound. The biocompatible polymer material is selected from one or more of polyvinylpyrrolidone, polyvinyl alcohol and methacrylated gelatin. The lappaconitine inclusion compound is a supramolecular inclusion compound formed by lappaconitine and cyclodextrin or cyclodextrin derivative. The cyclodextrin derivative is selected from at least one of sulfobutyl-beta-cyclodextrin, hydroxypropyl-beta-cyclodextrin sodium and methyl-beta-cyclodextrin. The microneedle containing the lappaconitine microneedle composition has the advantages of high drug loading, good safety, high transdermal efficiency, high mechanical strength and the like, and can improve the problem of strong local irritation of lappaconitine.

Description

Technical Field

[0001] This invention relates to the fields of biomedicine and transdermal drug delivery technology. More specifically, it relates to an aconitine microneedle composition, microneedles prepared therefrom, a preparation method therefrom, and their applications. Background Technology

[0002] Bulleyaconitine A is a natural Chinese herbal alkaloid used clinically to treat rheumatoid arthritis. Marketed formulations include tablets, oral solutions, and injections. It effectively repairs cartilage damage and synovial tissue destruction by reducing the expression of inflammatory factors and modulating the immune system, significantly treating rheumatoid arthritis. However, currently available oral formulations of bulleyaconitine suffer from tongue numbing sensation, low bioavailability, and short biological half-life. Injectable formulations also cause skin irritation and fail to address the short biological half-life issue, leading to poor patient compliance.

[0003] To address these issues, researchers have developed several new formulations of aconitine, including an aconitine injection powder published in Chinese Patent Publication No. CN100391457C. This formulation involves dissolving aconitine in a lactic acid solution, followed by isotonic dissolution, and then producing a lyophilized powder injection using freeze-drying. This method aims to reduce pain and irritation during injection. However, it has not improved patient compliance and ease of use, requiring precise injection by skilled technicians, which presents certain limitations and safety risks.

[0004] Microneedle technology, as a novel transdermal drug delivery system, can penetrate the stratum corneum barrier to deliver drugs directly to the dermis, achieving painless and highly effective local or systemic drug delivery. It can significantly improve the poor compliance issues associated with traditional aconitine formulations. However, aconitine is highly irritating locally and has poor water solubility. Therefore, maintaining high drug loading, good mechanical strength, and precise drug release characteristics while addressing its poor solubility and irritation is of great clinical significance. Summary of the Invention

[0005] Based on the above problems, the purpose of this invention is to provide an aconitine microneedle composition, microneedles prepared therefrom, a preparation method therefrom, and their applications. This aconitine microneedle composition, when used in microneedles, effectively improves the problem of strong local irritation caused by aconitine. Furthermore, it has been found that this technical solution can also improve the water solubility of aconitine. The resulting microneedles possess advantages such as high drug loading capacity, good safety, high transdermal efficiency, good mechanical strength, precise drug release, and good patient compliance.

[0006] To achieve the above objectives, the present invention adopts the following technical solution: In a first aspect, the present invention provides an aconitine microneedle composition, wherein the microneedle composition comprises, by mass percentage: 10-40% aconitine inclusion complex and 60-90% biocompatible polymer material; The biocompatible polymer material is selected from one or more of polyvinylpyrrolidone (PVP), polyvinyl alcohol (PVA), and methacrylamide gelatin (GelMA); The aconitin inclusion complex is: 1) Aconitum carmichaelii, and 2) Cyclodextrins or cyclodextrin derivatives The resulting supramolecular inclusion complex; The cyclodextrin derivative is selected from at least one of sulfobutyl-β-cyclodextrin, sodium hydroxypropyl-β-cyclodextrin, and methyl-β-cyclodextrin.

[0007] In the above-mentioned aconitine inclusion complex, 2) cyclodextrin or cyclodextrin derivatives are used as carrier materials to encapsulate 1) aconitine.

[0008] Aconitine itself has poor water solubility and strong irritation to the skin and mucous membranes, limiting its drug loading and transdermal absorption efficiency in microneedle formulations. The main objective of this invention is to improve the irritation of aconitine to the skin and mucous membranes. In this technical solution, aconitine is prepared into an aconitine inclusion complex. Utilizing the structural characteristics of the carrier (such as the hydrophobic cavities of cyclodextrin), aconitine molecules are effectively included or dispersed, significantly improving its apparent solubility in the aqueous phase and shielding its irritating functional groups, thus significantly reducing local skin adverse reactions. Furthermore, the complexation significantly improves the physicochemical stability of aconitine, preventing its degradation during microneedle preparation and storage. Experiments show that the aconitine inclusion complex has good compatibility with the biocompatible polymer material, enabling uniform dispersion of the aconitine inclusion complex while ensuring microneedle punctureability and significantly improving the drug loading and transdermal penetration efficiency of the microneedles.

[0009] Furthermore, the selection of specific cyclodextrins or cyclodextrin derivatives results in better transdermal effects and less irritation to the skin and mucous membranes when the microneedle composition is prepared.

[0010] In this technical solution, aconitine is included with cyclodextrin or cyclodextrin derivatives, so that the crystal form of aconitine in the resulting inclusion complex is changed to amorphous, which further avoids the irritation of aconitine in microneedles to the skin and mucous membranes, and at the same time improves the transdermal effect of aconitine in microneedles prepared from microneedles containing the microneedle composition.

[0011] Furthermore, the polyvinylpyrrolidone is selected from PVP K30 and / or PVP K90.

[0012] Furthermore, the mass ratio of aconitine to cyclodextrin or a cyclodextrin derivative is 1:0.5 to 1:4. Under these conditions, the aconitine-containing microneedles prepared from the microneedle composition have less irritation to the skin and mucous membranes.

[0013] Furthermore, the mass ratio of aconitine to cyclodextrin or cyclodextrin derivatives is 1:1 to 1:4.

[0014] Furthermore, the mass ratio of aconitine to cyclodextrin or cyclodextrin derivative is 1:1 to 1:2.

[0015] Furthermore, the preparation of the aconitin inclusion complex includes the following steps: Aconitum carmichaelii was dissolved in water with cyclodextrin or cyclodextrin derivatives, and the mixture was stirred and reacted in a water bath to obtain a clear solution. The solution was then freeze-dried to obtain the aconitine inclusion complex.

[0016] Furthermore, the temperature of the water bath is 30~50℃, the stirring speed is 700~1000r / min, and the time is 30~180min.

[0017] Furthermore, the freeze-drying temperature is -60 to -80°C.

[0018] Furthermore, the mass ratio of the aconitine inclusion complex to the biocompatible polymer material is 1:2 to 1:8. Microneedles prepared under these conditions possess high drug loading capacity, good safety, high transdermal efficiency, and good mechanical strength.

[0019] In a second aspect, the present invention provides a microneedle prepared from raw materials comprising the aconitine microneedle composition as described in the first aspect above and water.

[0020] The microneedles consist of a substrate and needles arranged in an array on the substrate.

[0021] Using the aforementioned raw materials, the microneedles can be prepared, and the resulting microneedles are integral needles.

[0022] Thirdly, the present invention provides a method for preparing microneedles as described in the second aspect above, comprising the following steps: A biocompatible polymer material is dissolved in water to obtain a matrix solution; Add the aconitine inclusion complex to the matrix solution and stir until completely dissolved to obtain a microneedle solution; The microneedle solution is added dropwise into a mold, dried under negative pressure, to obtain the microneedles.

[0023] Furthermore, the drying conditions are as follows: drying is carried out in a vacuum drying oven or a forced-air drying oven at a drying temperature of 20~35℃.

[0024] Furthermore, the vacuum degree of the vacuum drying is controlled at -0.08 to -0.1 MPa.

[0025] Furthermore, the biocompatible polymer material in the matrix solution has a mass percentage of 15-25%.

[0026] Furthermore, the negative pressure drying also includes a UV crosslinking step, wherein the UV crosslinking time is 1 to 5 minutes.

[0027] Fourthly, the present invention provides a microneedle patch comprising microneedles as described in the second aspect above and a backing bonded to the microneedles.

[0028] Fifthly, the present invention provides the use of the aconitine microneedle composition as described in the first aspect above, or the microneedles as described in the second aspect above, or the microneedle patches as described in the fourth aspect above, in the preparation of medicaments for the treatment and / or prevention of rheumatoid arthritis, osteoarthritis, or local analgesia.

[0029] The beneficial effects of this invention are as follows: In the technical solution of this invention, aconitine is loaded into microneedles in the form of a specific aconitine inclusion complex. Combined with a mild preparation process and optimized formula ratio, it achieves comprehensive technical effects such as significantly reducing the local skin irritation of aconitine, while possessing high aconitine solubility, excellent mechanical strength for insertion, efficient transdermal penetration and long-term stability. This significantly improves the local drug delivery effect and treatment compliance for patients with rheumatoid arthritis and other conditions. Attached Figure Description

[0030] The specific embodiments of the present invention will be described in further detail below with reference to the accompanying drawings.

[0031] Figure 1 The preparation method of the aconitine-sulfobutyl-β-cyclodextrin inclusion complex in Example 1 is shown.

[0032] Figure 2 The preparation method of aconitine inclusion complex microneedles is shown.

[0033] Figure 3 The FT-IR physical characterization results of the aconitine and sulfobutyl-β-cyclodextrin inclusion complex prepared in Example 1 are shown.

[0034] Figure 4 The results of DSC physical characterization of the aconitine and sulfobutyl-β-cyclodextrin inclusion complex prepared in Example 1 are shown.

[0035] Figure 5 The XRD physical characterization results of the aconitine and sulfobutyl-β-cyclodextrin inclusion complex prepared in Example 1 are shown.

[0036] Figure 6 The SEM physical characterization results of the aconitine and sulfobutyl-β-cyclodextrin inclusion complex prepared in Example 1 are shown.

[0037] Figure 7 The levels of IL-6 and TNF-α in blood samples from the aconitine inclusion complex microneedles prepared in Example 2 and other control groups are shown.

[0038] Figure 8 The microneedle-based microscopic morphology of the aconitine inclusion complex prepared in Example 1 is shown.

[0039] Figure 9 The mechanical properties test results of the aconitine inclusion complex microneedles prepared in Example 1 are shown.

[0040] Figure 10 The results show the joint and foot pad swelling rate, weight change, and overall score after treatment with the aconitine inclusion complex microneedles prepared in Example 1 and other control groups.

[0041] Figure 11 The Micro-CT scan results of the aconitine inclusion complex microneedles prepared in Example 1 and other control groups are shown.

[0042] Figure 12 The results of tests on trabecular bone thickness (Tb.Th), bone mineral density (BMD), bone surface area to volume ratio (BS / BV), and bone volume fraction (BV / TV) of the microneedles containing aconitine prepared in Example 1 and other control groups are shown.

[0043] Figure 13 The results of H&E staining and SO-FG staining experiments are shown for the microneedles of aconitine inclusion complex prepared in Example 1 and other control groups.

[0044] Figure 14 The results of immunohistochemical assays of the aconitine inclusion complex microneedles prepared in Example 1 and other control groups are shown.

[0045] Figure 15 The contents of IL-1β and IL-6 in the synovium and articular cartilage of the aconitine inclusion complex microneedles prepared in Example 1 and other control groups are shown.

[0046] Figure 16 The results of H&E staining of major organs after treatment are shown for the microneedles containing aconitine prepared in Example 1 and other control groups. Detailed Implementation

[0047] To more clearly illustrate the present invention, the following description, in conjunction with preferred embodiments and accompanying drawings, further explains the invention. Similar components in the drawings are indicated by the same reference numerals. Those skilled in the art should understand that the specific description below is illustrative rather than restrictive and should not be construed as limiting the scope of protection of the present invention.

[0048] Example 1 A microneedle composition containing aconitine inclusion complex is provided. The microneedle composition is composed of aconitine inclusion complex and microneedle matrix material in a mass ratio of 1:8. The aconitine inclusion complex is a supramolecular inclusion complex formed by aconitine and a cyclodextrin derivative. The mass ratio and selection of aconitine to cyclodextrin derivative in the aconitine inclusion complex, as well as the content of each component in the composition, are shown in Table 1 below.

[0049] A microneedle is prepared from a raw material comprising the microneedle composition described above, specifically including the following steps: (1) Preparation of microneedle matrix solution: Preparation of aconitine inclusion complex: According to the formula in Table 1, weigh aconitine and sulfobutyl-β-cyclodextrin in a mass ratio of 1:1, add an appropriate amount of purified water, and stir the mixture at 800 r / min under a 30℃ water bath for 90 minutes to obtain a clear aconitine / cyclodextrin inclusion complex solution. After freeze-drying, obtain the freeze-dried aconitine / cyclodextrin inclusion complex, i.e., the aconitine inclusion complex (e.g., ...). Figure 1 (As shown).

[0050] Prepare a matrix solution according to the microneedle matrix material formulation in Table 1, wherein the total mass content of the microneedle matrix material in the matrix solution is 22.5 wt%. The preparation method of the above matrix solution specifically includes: weighing the prescribed amount of injection water into a centrifuge tube, adding the prescribed amount of GelMA, stirring until completely dissolved, and obtaining the matrix solution.

[0051] (2) Preparation of microneedles: The prepared aconitine inclusion complex was mixed thoroughly with the matrix solution, and 80 μL was added dropwise to a 0.75 cm³ volume. 2 Apply the coating evenly to a mold of the desired size, apply negative pressure to the mold for about 15 minutes, then transfer the mold to a vacuum drying oven or forced-air drying oven for about 2 hours to obtain microneedles. After drying, the microneedles are cross-linked under ultraviolet light for 4 minutes before demolding to obtain microneedle patches (such as...). Figure 2 (As shown).

[0052] Table 1 Formulation of Aconitum carmichaelii inclusion complex microneedle composition

[0053] Example 2 A microneedle composition containing aconitine inclusion complex is provided. The microneedle composition is composed of aconitine inclusion complex and microneedle matrix material in a mass ratio of 1:8. The aconitine inclusion complex is a supramolecular inclusion complex formed by aconitine and cyclodextrin derivative. The mass ratio and selection of aconitine to cyclodextrin derivative in the aconitine inclusion complex, as well as the content of each component in the composition, are shown in Table 1.

[0054] The preparation method of microneedles and microneedle patches is the same as in Example 1, except that the mass ratio of aconitine to sulfobutyl-β-cyclodextrin is 1:2, and the total mass content of microneedle matrix material in the matrix solution is 20.0 wt%.

[0055] Example 3 A microneedle composition containing aconitine inclusion complex is provided. The microneedle composition is composed of aconitine inclusion complex and microneedle matrix material in a mass ratio of 1:8. The aconitine inclusion complex is a supramolecular inclusion complex formed by aconitine and cyclodextrin derivative. The mass ratio and selection of aconitine to cyclodextrin derivative in the aconitine inclusion complex, as well as the content of each component in the composition, are shown in Table 1.

[0056] The preparation method of microneedles and microneedle patches is the same as in Example 1, except that the mass ratio of aconitine to sulfobutyl-β-cyclodextrin is 1:3, and the total mass content of microneedle matrix material in the matrix solution is 25.0 wt%.

[0057] Example 4 A microneedle composition containing aconitine inclusion complex is provided. The microneedle composition is composed of aconitine inclusion complex and microneedle matrix material in a mass ratio of 1:8. The aconitine inclusion complex is a supramolecular inclusion complex formed by aconitine and cyclodextrin derivative. The mass ratio and selection of aconitine to cyclodextrin derivative in the aconitine inclusion complex, as well as the content of each component in the composition, are shown in Table 1.

[0058] The preparation method of microneedles and microneedle patches is the same as in Example 1, except that the mass ratio of aconitine to sulfobutyl-β-cyclodextrin is 1:4, and the total mass content of microneedle matrix material in the matrix solution is 22.5 wt%.

[0059] Example 5 A microneedle composition containing aconitine inclusion complex is provided. The microneedle composition is composed of aconitine inclusion complex and microneedle matrix material in a mass ratio of 1:8. The aconitine inclusion complex is a supramolecular inclusion complex formed by aconitine and cyclodextrin derivative. The mass ratio and selection of aconitine to cyclodextrin derivative in the aconitine inclusion complex, as well as the content of each component in the composition, are shown in Table 1.

[0060] The preparation method of microneedles and microneedle patches is the same as in Example 1, except that the mass ratio of aconitine to sulfobutyl-β-cyclodextrin is 2:1, and the total mass content of microneedle matrix material in the matrix solution is 18.0 wt%.

[0061] Comparative Example 1 A microneedle composition containing aconitine complex, wherein the microneedle composition is composed of aconitine and microneedle matrix material GelMA in a mass ratio of 1:8, and the specific formulation is shown in Table 1.

[0062] The preparation method of microneedles and microneedle patches is the same as in Example 1, except that cyclodextrin inclusion is not used, and aconitine powder is directly dispersed in GelMA aqueous solution, and the total mass content of GelMA in GelMA aqueous solution is 22.5 wt%.

[0063] Comparative Example 2 A microneedle composition containing aconitine inclusion complex, wherein the aconitine inclusion complex is a supramolecular inclusion complex formed by aconitine and a cyclodextrin derivative, and the mass ratio and selection of aconitine to cyclodextrin derivative in the aconitine inclusion complex, as well as the content of each component in the composition, are shown in Table 1.

[0064] The preparation method of microneedles and microneedle patches is the same as in Example 1, except that the polymer material GelMA is not used, and only the aconitine / cyclodextrin inclusion complex solution is used to prepare microneedles.

[0065] The test results of Examples 1-5 and Comparative Examples 1-2 are shown in Table 2 below. In all examples, aconitine can form stable inclusion complexes with cyclodextrin, and the apparent solubility is 30-70 times higher than that of the original drug (Comparative Example 1). The total impurities of related substances in all examples are controlled at 0.5%-1.0%, and the microneedles have good morphology. However, in Comparative Example 1, no cyclodextrin inclusion complex was used, and aconitine was almost insoluble in water, the solution was turbid and there was precipitation, the drug loading of the microneedles was uneven, and it could not meet the drug administration requirements. Moreover, the total impurities were as high as 2.7%, indicating that its stability was insufficient. In addition, Comparative Example 2 did not add the polymer material GelMA, and it could not form needles by relying solely on the inclusion complex itself, which shows the necessity of polymer materials for microneedle formation.

[0066] Table 2. Detection results of related substances in microneedles prepared from aconitine inclusion complex microneedles.

[0067] Physical characterization of aconitine cyclodextrin inclusion complex: The aconitine inclusion complex prepared in Example 1 was subjected to Fourier transform infrared spectroscopy (FT-IR) for physical characterization. Free aconitine, sulfobutyl-β-cyclodextrin, the aconitine / sulfobutyl-β-cyclodextrin inclusion complex, and physical mixtures were analyzed. The sample was thoroughly mixed with dry potassium bromide powder in an agate mortar and finely ground. Subsequently, a portion of the homogenized powder was placed in a tableting mold and compressed using a hydraulic press at a pressure of 108 MPa to form transparent tablets. Finally, the tablets were heated at 400–4000 cm⁻¹. -1 FT-IR spectra were recorded within the wavenumber range. Fourier transform infrared spectroscopy (FT-IR) technology, by detecting vibrational changes in characteristic functional groups, can confirm whether drug molecules are included in cyclodextrin cavities, thus distinguishing inclusion compounds from physical mixtures. Figure 3 It can be seen from this that aconitine is present at 3520 cm. -1 (N–H stretching vibration), 2927 cm -1 (C–H stretching), 1300–1000 cm -1 (C–O stretching), 1617 cm -1 (C=C stretching) and 1727 cm -1 Characteristic absorption peaks are observed at (C=O stretching, characteristic peaks of esters). In the spectrum of sulfonyl-β-cyclodextrin, a characteristic absorption peak is visible at 3445 cm⁻¹. -1 A broad absorption band generated by the stretching vibration of free O–H was observed at 2927 cm⁻¹, and a band at 2927 cm⁻¹ was also visible. -1 C–H stretching vibration at 1660 cm -1 C=O stretching vibration at 1197 cm -1 C–O stretching vibration at 1031 cm -1 The C–O–C stretching vibration at 3445 cm⁻¹. The spectrum of aconitine / sulfobutyl-β-cyclodextrin at 3445 cm⁻¹. -1 A broad absorption band remarkably similar to that of sulfobutyl-β-cyclodextrin was observed at this region, while the N–H stretching peak of BLA was absent. Furthermore, aconitine showed an absorption band at 1727 cm⁻¹. -1 and 1617 cm -1 The absorption peak at that location was significantly weaker than that in the physical mixture, which further confirms the formation of the inclusion compound.

[0068] The aconitine inclusion complex prepared in Example 1 was used for physical characterization by differential scanning calorimetry (DSC). A certain amount of sample was uniformly placed in an aluminum crucible, sealed using a dedicated tablet press, and placed under a nitrogen atmosphere (flow rate of 100 mL / min). The analysis was performed using a DSC. DSC measurements were conducted on free aconitine, sulfobutyl-β-cyclodextrin, the aconitine / sulfobutyl-β-cyclodextrin inclusion complex, and physical mixtures. The sample was heated from 30°C to 400°C at a heating rate of 10°C / min. DSC was used to confirm the formation of the inclusion complex by evaluating differences in crystallization state, melting point, and phase transition behavior. Figure 4 As can be seen, aconitine exhibits an endothermic peak at 165℃, which is still present in the physical mixture of aconitine and sulfobutyl-β-cyclodextrin. In contrast, the characteristic endothermic peak disappears in the DSC curve of aconitine / sulfobutyl-β-cyclodextrin, indicating that its thermal behavior has changed, thus confirming the formation of cyclodextrin inclusion complex in an amorphous, non-crystalline form.

[0069] The aconitine inclusion complex prepared in Example 1 was used for physical characterization by X-ray diffraction analysis. An appropriate amount of sample powder was evenly spread on the sample stage, compacted, and excess powder was removed to ensure a smooth surface. Using an X-ray diffractometer (XRD) equipped with a copper target (wavelength = 1.5406 Å), diffraction data were collected within the 2θ angle range of 0.2° to 120°. The obtained diffraction patterns were analyzed, and crystal phase transitions were identified by comparing peak positions and intensities. X-ray diffraction (XRD) ruled out simple physical mixing by detecting changes in crystal structure, as both sulfobutyl-β-cyclodextrin and aconitine exhibit characteristic crystalline structures. Figure 5 As can be seen, the XRD diffraction pattern of aconitine shows multiple strong and sharp diffraction peaks at diffraction angles (2θ) of 7.75°, 9.36°, 17.81°, and 18.68°, confirming the crystalline nature of the aconitine powder. The XRD diffraction pattern of the physical mixture shows similar sharp diffraction peaks to aconitine, indicating that no interaction occurred between the components. Conversely, the XRD diffraction patterns of sulfobutyl-β-cyclodextrin and the aconitine / sulfobutyl-β-cyclodextrin inclusion complex are relatively smooth, without crystalline diffraction peaks. This proves that aconitine lost its crystalline state during the inclusion process, and the inclusion complex formed and exhibited an amorphous morphology.

[0070] The aconitine inclusion complex prepared in Example 1 was used for scanning electron microscopy (SEM) to observe changes in its physical morphology. Free aconitine, sulfobutyl-β-cyclodextrin, the aconitine / sulfobutyl-β-cyclodextrin inclusion complex, and physical mixtures were analyzed. A small amount of sample powder was uniformly dispersed on a carbon ribbon and then observed using a scanning electron microscope to study the surface morphology of free aconitine, sulfobutyl-β-cyclodextrin, the aconitine / sulfobutyl-β-cyclodextrin inclusion complex, and physical mixtures. Image acquisition was performed in a vacuum environment at a magnification of 500×, and the operation was completed using SEM control software. Using scanning electron microscopy (SEM) to observe the surface morphology changes after cyclodextrin encapsulation allows direct observation of the morphological reconstruction process, rather than the coexistence state of two independent substances. Figure 6 As can be seen, at 500x magnification, sulfobutyl-β-cyclodextrin appears as spherical particles of varying sizes. The morphology of this physical mixture is similar to that of sulfobutyl-β-cyclodextrin, which is consistent with the characteristics of a mixture formed by grinding sulfobutyl-β-cyclodextrin and aconitine, rather than an inclusion complex. In contrast, the aconitine / sulfobutyl-β-cyclodextrin inclusion complex exhibits an irregular, plate-like morphology, which is distinctly different from that of sulfobutyl-β-cyclodextrin. This indicates that there is an intermolecular interaction between aconitine and sulfobutyl-β-cyclodextrin, which alters their morphology and leads to the formation of a new substance.

[0071] Skin irritation test: Skin irritation tests were conducted using the aconitine inclusion complex microneedles prepared in Examples 1-5 and Comparative Example 1. Six healthy New Zealand white rabbits weighing 2.0-2.5 kg were selected. Twenty-four hours prior to the experiment, the rabbits were shaved on both sides of their spine, with a shaved area of ​​approximately 3 cm × 3 cm, and the skin was checked to confirm no damage. The experimental animals were randomly divided into six groups, one rabbit per group. The microneedle patch to be tested was applied to the left shaved area, while the right shaved area served as a blank control, with only a blank GelMA microneedle patch without the drug applied. The microneedle patch was applied to the shaved area on the rabbit's back and secured with medical tape. A force of 70 N / cm² was applied for 20 seconds to ensure complete microneedle penetration into the skin. The microneedle patch was removed 1 hour after application, and skin reactions were observed and recorded immediately after removal (0 h), 1 h, 24 h, and 48 h.

[0072] The results of the aconitine microneedle skin irritation test are shown in Table 3.

[0073] Table 3. Skin irritation test of Aconitum carmichaelii microneedles

[0074] Example 6 A microneedle composition containing aconitine inclusion complex is provided. The microneedle composition is composed of aconitine inclusion complex and microneedle matrix material in a mass ratio of 1:8. The aconitine inclusion complex is a supramolecular inclusion complex formed by aconitine and sulfobutyl-β-cyclodextrin. The mass ratio and selection of aconitine to sulfobutyl-β-cyclodextrin in the aconitine inclusion complex, as well as the content of each component in the composition, are shown in Table 4.

[0075] The preparation method of microneedles and microneedle patches is the same as in Example 1, except that the type of microneedle matrix material is different, and the microneedle matrix material is polyvinyl alcohol (PVA).

[0076] Table 4 Formulation of Aconitum carmichaelii inclusion complex microneedle composition

[0077] Example 7 A microneedle composition containing aconitine inclusion complex is provided. The microneedle composition is composed of aconitine inclusion complex and microneedle matrix material in a mass ratio of 1:8. The aconitine inclusion complex is a supramolecular inclusion complex formed by aconitine and sulfobutyl-β-cyclodextrin. The mass ratio and selection of aconitine to sulfobutyl-β-cyclodextrin in the aconitine inclusion complex, as well as the content of each component in the composition, are shown in Table 4.

[0078] The preparation method of microneedles and microneedle patches is the same as in Example 1, except that the type of microneedle matrix material is different, and the microneedle matrix material is polyvinylpyrrolidone K30 (PVPK30).

[0079] Example 8 A microneedle composition containing aconitine inclusion complex is provided. The microneedle composition is composed of aconitine inclusion complex and microneedle matrix material in a mass ratio of 1:8. The aconitine inclusion complex is a supramolecular inclusion complex formed by aconitine and sulfobutyl-β-cyclodextrin. The mass ratio and selection of aconitine to sulfobutyl-β-cyclodextrin in the aconitine inclusion complex, as well as the content of each component in the composition, are shown in Table 4.

[0080] The preparation method of microneedles and microneedle patches is the same as in Example 1, except that the type of microneedle matrix material is different, and the microneedle matrix material is polyvinylpyrrolidone K90 (PVPK90).

[0081] Comparative Example 3 A microneedle composition containing aconitine inclusion complex is shown in Table 4.

[0082] The preparation method of microneedles and microneedle patches is the same as in Example 1, except that the microneedle matrix material is mannitol, a small molecule excipient, instead of GelMA.

[0083] Comparative Example 4 A microneedle composition containing aconitine inclusion complex is shown in Table 4.

[0084] The preparation method of microneedles and microneedle patches is the same as in Example 1, except that the microneedle matrix material is sucrose, a small molecule excipient, instead of GelMA.

[0085] Skin puncture test: Skin puncture tests were conducted using the aconitine inclusion complex microneedles prepared in Examples 6-8 and Comparative Examples 3-4. Fresh Bama miniature pig skin, preserved at -20°C, was removed and cut to an appropriate size. After thawing naturally at room temperature, excess moisture was blotted dry with lint-free paper. The skin was then laid flat on the experimental table with the stratum corneum facing upwards. The microneedle to be tested was placed on the pig skin, with the needle body facing the stratum corneum, and the delivery device was used at a speed of 70 N / cm. 2 Press the microneedles firmly for 20 seconds, then remove the delivery device, peel off the residual patch, stain with methylene blue solution (0.5wt%), wipe off the excess methylene blue solution after 1 hour, observe and count the number of blue holes formed in the microneedle array under a stereomicroscope, and calculate the insertion success rate.

[0086] The success rate (%) of the insertion is calculated as follows: (Number of microneedles forming blue holes / Total number of microneedles) × 100%. The success rate of microneedle skin insertion of aconitine inclusion complex is shown in Table 5.

[0087] Table 5 Success rate of microneedle skin penetration of aconitine inclusion complex

[0088] The success rate of skin insertion of aconitine inclusion complex microneedle compositions depends on the presence of polymeric materials in the microneedle matrix solution. Formulations without polymeric materials can cause the microneedle patch to become severely hygroscopic or brittle, preventing complete peeling and leading to needle breakage or melting. Therefore, the presence of polymeric materials is necessary.

[0089] Skin irritation test: Skin irritation tests were conducted using the aconitine inclusion complex microneedles prepared in Examples 6-8. Three healthy New Zealand white rabbits weighing 2.0-2.5 kg were selected. Twenty-four hours prior to the experiment, the rabbits were shaved on both sides of their spine, with a shaved area of ​​approximately 3 cm × 3 cm, and the skin was checked to confirm no damage. The experimental animals were randomly divided into three groups, one rabbit per group. The microneedle patch to be tested was applied to the left shaved area, while the right shaved area served as a blank control, with only a blank GelMA microneedle patch without the drug applied. The microneedle patch was applied to the shaved area on the rabbit's back and secured with medical tape. A force of 70 N / cm² was applied for 20 seconds to ensure complete microneedle penetration into the skin. The microneedle patch was removed 1 hour after application, and skin reactions were observed and recorded immediately after removal (0 h), 1 h, 24 h, and 48 h.

[0090] The results of the skin irritation test of aconitine microneedles are shown in Table 6.

[0091] Table 6. Skin Irritation Test of Aconitum carmichaelii Microneedle

[0092] Example 9 A microneedle composition containing aconitine inclusion complex is provided. The microneedle composition is composed of aconitine inclusion complex and microneedle matrix material in a mass ratio of 1:8. The aconitine inclusion complex is a supramolecular inclusion complex formed by aconitine and cyclodextrin derivative. The mass ratio and selection of aconitine to cyclodextrin derivative in the aconitine inclusion complex, as well as the content of each component in the composition, are shown in Table 7.

[0093] The preparation method of microneedles and microneedle patches is the same as in Example 1, except that the type of cyclodextrin is different, and the type of cyclodextrin is hydroxypropyl-β-cyclodextrin (HP-β-CD), and the mass ratio of aconitine to cyclodextrin is 1:1.

[0094] Table 7 Formulation of Aconitum carmichaelii inclusion complex microneedle composition

[0095] Example 10 A microneedle composition containing aconitine inclusion complex is provided. The microneedle composition is composed of aconitine inclusion complex and microneedle matrix material in a mass ratio of 1:8. The aconitine inclusion complex is a supramolecular inclusion complex formed by aconitine and cyclodextrin derivative. The mass ratio and selection of aconitine to cyclodextrin derivative in the aconitine inclusion complex, as well as the content of each component in the composition, are shown in Table 7.

[0096] The preparation method of microneedles and microneedle patches is the same as in Example 1, except that the type of cyclodextrin is different, and the type of cyclodextrin is methyl-β-cyclodextrin (Me-β-CD), and the mass ratio of aconitine to cyclodextrin derivative is 1:1.

[0097] Example 11 A microneedle composition containing aconitine inclusion complex is provided. The microneedle composition is composed of aconitine inclusion complex and microneedle matrix material in a mass ratio of 1:3. The aconitine inclusion complex is a supramolecular inclusion complex formed by aconitine and cyclodextrin derivative. The mass ratio and selection of aconitine to cyclodextrin derivative in the aconitine inclusion complex, as well as the content of each component in the composition, are shown in Table 7.

[0098] The preparation method of microneedles and microneedle patches is the same as in Example 1, except that the mass ratio of aconitine to cyclodextrin derivative is different, and the mass ratio of aconitine to cyclodextrin derivative is 1:4.

[0099] Comparative Example 5 A microneedle composition containing aconitine inclusion complex is shown in Table 7.

[0100] The preparation method of microneedles and microneedle patches is the same as in Example 1, except that cyclodextrin is not used to form aconitine inclusion complex, and aconitine is dissolved in anhydrous ethanol.

[0101] Comparative Example 6 A microneedle composition containing aconitine inclusion complex is shown in Table 7.

[0102] The preparation method of microneedles and microneedle patches is the same as in Example 1, except that cyclodextrin is not used to form aconitine inclusion complex, and aconitine is dissolved in polyethylene glycol 400 (PEG400).

[0103] Comparative Example 7 A microneedle composition containing aconitine inclusion complex is shown in Table 7.

[0104] The preparation method of microneedles and microneedle patches is the same as in Example 1, except that cyclodextrin is not used to form aconitine inclusion complex, and aconitine is encapsulated into microspheres with polylactic acid-glycolic acid copolymer (PLGA).

[0105] Comparative Example 8 A microneedle composition containing aconitine inclusion complex is shown in Table 7.

[0106] The preparation method of microneedles and microneedle patches is the same as in Example 1, except that instead of using specific cyclodextrins or cyclodextrin derivatives to form aconitine inclusion complexes, quaternized β-cyclodextrin (QA-β-CD) is used to encapsulate aconitine to form aconitine inclusion complexes.

[0107] Skin irritation test: Skin irritation tests were conducted using the aconitine inclusion complex microneedles prepared in Examples 9-11 and Comparative Examples 5-8. Five healthy New Zealand white rabbits weighing 2.0-2.5 kg were selected. Twenty-four hours prior to the experiment, the rabbits were shaved on both sides of their spine, with a shaved area of ​​approximately 3 cm × 3 cm, and the skin was checked to confirm no damage. The experimental animals were randomly divided into five groups, one rabbit per group. The microneedle patch to be tested was applied to the left shaved area, while the right shaved area served as a blank control, with only a blank GelMA microneedle patch without the drug applied. The microneedle patch was applied to the shaved area on the rabbit's back and secured with medical tape. A force of 70 N / cm² was applied for 20 seconds to ensure complete microneedle penetration into the skin. The microneedle patch was removed 1 hour after application, and skin reactions were observed and recorded immediately after removal (0 h), 1 h, 24 h, and 48 h.

[0108] The results of the skin irritation test of aconitine microneedling are shown in Table 8.

[0109] Table 8. Skin irritation test of Aconitum carmichaelii microneedles

[0110] Aconitum carmichaelii has poor water solubility. When added directly to the microneedle matrix solution, it forms a poorly soluble white precipitate. However, when dissolved in an organic solvent, it can be uniformly mixed in the microneedle matrix solution. But the microneedles prepared from this solution are more irritating to the skin than the aconitine / cyclodextrin complex microneedles. Therefore, when loading aconitine into microneedles, it is necessary to first modify it into an aconitine inclusion complex using a specific cyclodextrin or cyclodextrin derivative.

[0111] In vitro delivery efficiency test: In vitro delivery efficiency tests were conducted using microneedles dissolved with the aconitine inclusion complexes prepared in Examples 9-11 and Comparative Examples 5-8. Fresh Bama miniature pig skin, preserved at -20°C, was removed and cut to an appropriate size. After thawing naturally at room temperature, excess moisture was blotted dry with lint-free paper. The skin was then laid flat on the experimental table with the stratum corneum facing upwards. The microneedles to be tested were placed on the pig skin, with the needle body facing the stratum corneum, and the delivery device was used at a speed of 70 N / cm. 2 Press the microneedle firmly for 20 seconds, then remove the drug delivery device. After 1 hour, peel off the residual patch and collect the residual adhesive on the pigskin. Collect the residue into a centrifuge tube and use HPLC to detect the residual drug content.

[0112] Wherein, the amount of drug entering the skin = the amount of drug in the microneedle - the amount of drug remaining in the microneedle; delivery efficiency (%) = the amount of drug entering the skin / the amount of drug in the microneedle × 100%.

[0113] The delivery efficiency of teriparatide dissolving microneedles is shown in Table 9.

[0114] Table 9. Results of delivery efficiency test for the swollen microneedle composition

[0115] Encapsulation of aconitine with cyclodextrin derivatives can significantly improve the efficiency of transdermal drug delivery via microneedles. The inclusion of aconitine with the cyclodextrin derivatives selected in this invention alters its crystal form to an amorphous state, facilitating skin penetration and thus improving microneedle delivery efficiency and bioavailability. Microneedle compositions without the cyclodextrin derivatives selected in this invention cannot achieve efficient drug delivery; therefore, the presence of these cyclodextrin derivatives is necessary.

[0116] Effects of aconitine inclusion complex microbes on serum inflammatory factor expression levels: In vivo pharmacodynamic studies were conducted using the aconitine inclusion complex microneedles prepared in Example 1 and commercially available aconitine tablets.

[0117] In vivo pharmacodynamic experiments were conducted using female SD rats. Twelve female SD rats were randomly divided into four groups of three rats each, and each group was labeled. The day before the experiment, leg hair was removed from the rats and treated with depilatory cream. Control group: No medication administered; normal feeding. Model group: Rheumatoid arthritis (CIA) model established; no medication administered. Oral administration group: Rheumatoid arthritis (CIA) model established; dosage was 13.6 μg / rat. Aconitum carmichaelii inclusion complex microneedle administration group: Rheumatoid arthritis (CIA) model established; dosage was 13.6 μg / rat. The rats were held down on a table, the microneedle patch packaging was opened, the blister pack was torn off, the patch was removed, and the release paper was peeled off. The adhesive portion of the microneedle was applied to the leg, and the microneedle was immediately pressed with the thumb for 20 seconds. Medical tape was then wrapped around the rat's leg. The patch was removed from the skin after 24 hours. After 21 days of continuous administration, blood was collected from the eyeballs. The whole blood was collected in centrifuge tubes and centrifuged at 2000 rpm for 10 min at 4°C. The supernatant serum was collected by pipette, and the expression levels of inflammatory factors IL-6 and TNF-α in the blood samples were detected using an ELISA kit.

[0118] In vivo pharmacodynamic test results The therapeutic effect of aconitine inclusion complex microbes on CIA rats was studied by comparing them with the control group, model group, and oral administration. The expression levels of inflammatory factors IL-6 and TNF-α in the blood samples of rats from different groups were compared. Figure 7 As shown.

[0119] from Figure 7 It can be seen that both microneedle administration of aconitine inclusion complex and oral administration of aconitine have therapeutic effects on CIA rats, and can significantly reduce the expression levels of inflammatory factors IL-6 and TNF-α in rat serum. Among them, the microneedle administration group has better therapeutic effects and the lowest expression of inflammatory factors in rat serum.

[0120] Microneedle mechanical strength test: The mechanical strength of the microneedles was tested using the aconitine inclusion complex microneedles prepared in Example 1.

[0121] The mechanical properties of MNs were evaluated using a force-displacement testing machine. MNs were attached to the stage with double-sided tape, needle tip facing upwards; a displacement rate of 1.1 mm / s was applied; the force measurement limit was set to 5 N; stress changes and displacement data of MNs were recorded every 0.01 seconds to generate stress-strain curves for MNs. After compression, the morphological changes of MNs were observed using a stereomicroscope.

[0122] Microneedle mechanical strength test results Figure 8 The morphology of aconitine inclusion complex microneedles was shown under a stereomicroscope. The microneedles had an intact structure. Figure 9 As can be seen, the force-displacement curves did not show significant bending or breakage throughout the entire test range, indicating that the aconitine cyclodextrin microneedle samples maintained structural integrity and possessed sufficient mechanical strength to withstand the forces exerted during skin penetration. When the microneedles were compressed to 70% deformation, the maximum axial force of a single microneedle was 2.677 N. Previous studies have confirmed that a single microneedle capable of withstanding an axial force exceeding 0.15 N can successfully penetrate the skin. Based on this standard, the prepared microneedles exhibit excellent mechanical strength and satisfactory structural integrity, demonstrating superior mechanical properties and enabling effective transdermal drug delivery.

[0123] The therapeutic effects of the micro-targeted therapy on CIA rats were evaluated using joint swelling assessment and clinical scoring: The aconitine inclusion complex microneedles prepared in Example 1 and commercially available aconitine tablets were used to treat CIA rats. The treatment effect was then observed by assessing joint swelling and clinical scoring.

[0124] Thirty female SD rats were randomly divided into five groups of six rats each and marked. The day before the experiment, leg hair was removed from the rats and treated with depilatory cream. Control group: No medication administered; normal feeding. Model group: Rheumatoid arthritis (CIA) model established in rats; no medication administered. Blank microneedle treatment group: CIA model established in rats; treated with blank microneedles; no medication administered. Oral administration group: CIA model established in rats; dose of medication was 13.6 μg / rat. Microneedle administration group of aconitine inclusion complex: A rat model of rheumatoid arthritis (CIA) was established. The dosage was 13.6 μg / rat. The rats were held on a table, the outer packaging of the microneedle patch was opened, the blister pack was torn off, the patch was removed, and the release paper was peeled off. The adhesive portion of the microneedle edge was applied to the leg, and the microneedle was immediately pressed with the thumb for 20 seconds. Then, medical tape was wrapped around the rat's leg. The patch was removed from the skin after 24 hours. The degree of joint swelling in the rats was assessed using a composite scoring system: each limb was scored separately, and the total score was the sum of the scores for all four limbs. The scoring criteria were divided into five levels (0 points: no erythema or swelling; 1 point: mild erythema and swelling limited to the tarsal joint or ankle joint; 2 points: moderate erythema and swelling extending from the ankle joint to the metatarsophalangeal joint; 3 points: severe erythema and swelling extending from the metatarsophalangeal joint to the toe; 4 points: severe erythema and swelling involving the ankle joint, claw and toe, or limb rigidity)

[29] . An assessment was conducted every 3 days; at the same time, the diameter of the claw and the diameter of the ankle joint were measured every 3 days using calipers, and the weight changes were recorded for 21 days.

[0125] Joint swelling assessment and clinical scoring results Except for the normal control group, all other groups experienced joint and footpad swelling after immunization. Figure 10 As shown, after 21 days of treatment, the swelling of the right hind limb of rats in different groups varied. The swelling of the right hind limb of rats treated with oral aconitine and aconitine inclusion complex microneedling was reduced. The aconitine inclusion complex microneedling treatment group showed the most significant swelling reduction. Both the aconitine inclusion complex microneedling group and the oral aconitine group could continuously reduce the joint injury score of CIA rats and reduce the degree of hind limb swelling. The aconitine inclusion complex microneedling group had the most obvious treatment effect and minimized the impact on the weight gain of rats.

[0126] Observation of the therapeutic effect on CIA rats using Micro-CT: The aconitine inclusion complex microneedles prepared in Example 1 and commercially available aconitine tablets were used to treat CIA rats, and the treatment effect was then observed by Micro-CT.

[0127] Thirty female SD rats were randomly divided into five groups of six rats each and marked. The day before the experiment, leg hair was removed from the rats and treated with depilatory cream. Control group: No medication administered; normal feeding. Model group: Rheumatoid arthritis (CIA) model established in rats; no medication administered. Blank microneedle treatment group: CIA model established in rats; treated with blank microneedles; no medication administered. Oral administration group: CIA model established in rats; dose of medication was 13.6 μg / rat. Microneedle administration group of aconitine inclusion complex: A rat model of rheumatoid arthritis (CIA) was established. The dosage was 13.6 μg / rat. The rats were held on a table, the outer packaging of the microneedle patch was opened, the blister pack was torn off, the microneedle patch was removed, the release paper was peeled off, and the adhesive part of the microneedle edge was first applied to the leg. The microneedle was immediately pressed with the thumb for 20 seconds, and then the rat's leg was wrapped with medical tape. The patch was peeled off from the skin after 24 hours. After 21 days of continuous administration, the right hind limb of the rats was taken for Micro-CT scan to observe the treatment effect of different groups, and the data related to the trabecular bone of the rat hind limb ankle joint were analyzed.

[0128] Micro-CT trial results The therapeutic effect of aconitine inclusion complex microbes on CIA rats was studied by comparing them with the control group, model group, and oral administration group. The CT scans of the right hind limbs of rats in different groups were compared. Figure 11 As shown, the data on trabecular bone thickness (Tb.Th), bone mineral density (BMD), bone surface area to volume ratio (BS / BV), and bone volume fraction (BV / TV) in the right hind limb ankle joint of different groups of rats are compared as follows: Figure 12 As shown.

[0129] from Figure 11 As can be seen from the data, compared with the CIA model group and the blank microneedling treatment group, both the aconitine inclusion complex microneedling group and the oral aconitine group showed partial repair of ankle bone destruction, with the aconitine inclusion complex microneedling group showing significantly better repair effects. Figure 12 As can be seen, both the CIA model group and the blank microneedle treatment group showed significant reductions in BV / TV, Tb.Th, and BMD, while the BS / BV ratio increased, indicating a sparse trabecular network and damaged bone structure. After treatment with aconitine, all of the above bone structure parameters were significantly improved, with the aconitine inclusion complex microneedle group showing a more significant protective effect. This may be attributed to the excellent transdermal performance of the microneedle delivery system and its local administration to the hindlimb, which achieves higher bioavailability compared to oral administration. These results confirm that aconitine inclusion complex microneedles can achieve excellent drug delivery efficiency and bioavailability, providing a more advantageous treatment option for the long-term treatment of rheumatoid osteoarthritis.

[0130] The repair of ankle joints and synovium in CIA rats was observed using H&E staining and SO-FG staining: The aconitine inclusion complex microneedles prepared in Example 1 and commercially available aconitine tablets were used to treat CIA rats. The repair of ankle joints and synovium in CIA rats was then observed using hematoxylin-eosin (H&E) and saffron O-fast green (SO-FG).

[0131] Thirty female SD rats were randomly divided into five groups of six rats each and marked. The day before the experiment, leg hair was removed from the rats and treated with depilatory cream. Control group: No medication administered; normal feeding. Model group: Rheumatoid arthritis (CIA) model established in rats; no medication administered. Blank microneedle treatment group: CIA model established in rats; treated with blank microneedles; no medication administered. Oral administration group: CIA model established in rats; dose of medication was 13.6 μg / rat. Microneedle administration group of aconitine inclusion complex: A rat model of rheumatoid arthritis (CIA) was established. The dosage was 13.6 μg / rat. The rats were held on a table, the outer packaging of the microneedle patch was opened, the blister pack was torn off, the microneedle patch was removed, the release paper was peeled off, and the adhesive part of the microneedle edge was first applied to the leg. The microneedle was immediately pressed with the thumb for 20 seconds, and then the rat's leg was wrapped with medical tape. The patch was peeled off from the skin after 24 hours. After 21 days of continuous administration, the ankle joint of the right hind limb of the rat was isolated, and the pathological changes of the synovium and cartilage tissue were observed by H&E staining and SO-FG staining.

[0132] Results of H&E staining and SO-FG staining experiments from Figure 13As can be seen from H&E staining, severe bone damage, extensive inflammatory cell infiltration, synovial hyperplasia, cartilage degeneration, and bone erosion were observed in both the CIA model group and the blank microneedle group. After oral administration of aconitine combined with microneedle treatment, synovial inflammation was relieved, damaged cartilage was repaired, and the joint cavity structure returned to its normal morphology. Among them, the pathological damage in the aconitine inclusion complex microneedle administration group was significantly lower than that in the oral aconitine group. Safranin O-Fast Green (SO-FG) staining results further confirmed that the aconitine inclusion complex microneedle delivery system can more effectively maintain the integrity of the articular cartilage matrix. These results confirm that local application of aconitine inclusion complex microneedles can produce the most significant osteoprotective and cartilage repair effects. This enhanced efficacy is attributed to the microneedle-mediated direct skin penetration to the joint lesion site—a method that bypasses systemic circulation, thereby achieving stronger targeting, higher bioavailability, and faster onset of action, which is superior to oral administration. Furthermore, the aconitine inclusion complex system enables sustained drug release after skin implantation, which helps to improve the local inflammatory microenvironment in the long term and enhances the protection and repair of bone tissue. This method is highly suitable for the clinical needs of long-term treatment intervention in patients with rheumatoid arthritis.

[0133] Immunohistochemical observation of the effects of treatment on the expression levels of inflammatory factors in the ankle joint and synovium of CIA rats: The aconitine inclusion complex microneedles prepared in Example 1 and commercially available aconitine tablets were used to treat CIA rats. Immunohistochemistry was then used to observe the effects of treatment on the expression levels of inflammatory factors in the ankle joint and synovium of CIA rats.

[0134] Thirty female SD rats were randomly divided into five groups of six rats each and marked. The day before the experiment, leg hair was removed from the rats and treated with depilatory cream. Control group: No medication administered; normal feeding. Model group: Rheumatoid arthritis (CIA) model established in rats; no medication administered. Blank microneedle treatment group: CIA model established in rats; treated with blank microneedles; no medication administered. Oral administration group: CIA model established in rats; dose of medication was 13.6 μg / rat. Microneedle administration group of aconitine inclusion complex: A rat model of rheumatoid arthritis (CIA) was established. The dosage was 13.6 μg / rat. The rats were held on a table, the outer packaging of the microneedle patch was opened, the blister pack was torn off, the microneedle patch was removed, the release paper was peeled off, and the adhesive part of the microneedle edge was first applied to the leg. The microneedle was immediately pressed with the thumb for 20 seconds, and then the rat's leg was wrapped with medical tape. The patch was peeled off from the skin after 24 hours. After 21 days of continuous administration, the ankle joint of the right hind limb of the rats was isolated, and the changes in the expression levels of inflammatory factors in the ankle joint and synovium of CIA rats before and after treatment were observed by immunohistochemistry.

[0135] Immunohistochemical results from Figure 14-15 The results showed that, compared with the control group, the model group exhibited significantly increased IL-1β and IL-6 expression levels in both synovium and cartilage tissue; while the cytokine expression levels in the blank microneedle group were comparable to those in the model group. In both the oral aconitine treatment group and the aconitine inclusion complex microneedle treatment group, the protein levels of IL-1β and IL-6 in synovium and cartilage tissue were reduced, with the aconitine inclusion complex microneedle treatment group showing a more significant therapeutic effect.

[0136] In vivo safety evaluation: The aconitine inclusion complex microneedles prepared in Example 1 and commercially available aconitine tablets were used to treat CIA rats. The in vivo safety of the drugs was then assessed by H&E staining of major organ sections.

[0137] Thirty female SD rats were randomly divided into five groups of six rats each and marked. The day before the experiment, leg hair was removed from the rats and treated with depilatory cream. Control group: No medication administered; normal feeding. Model group: Rheumatoid arthritis (CIA) model established in rats; no medication administered. Blank microneedle treatment group: CIA model established in rats; treated with blank microneedles; no medication administered. Oral administration group: CIA model established in rats; dose of medication was 13.6 μg / rat. Microneedle administration group of aconitine inclusion complex: A rat model of rheumatoid arthritis (CIA) was established. The dosage was 13.6 μg / rat. The rats were held on a table, the outer packaging of the microneedle patch was opened, the blister pack was torn off, the microneedle patch was removed, the release paper was peeled off, and the adhesive part of the microneedle edge was first applied to the leg. The microneedle was immediately pressed with the thumb for 20 seconds, and then the rat's leg was wrapped with medical tape. The patch was peeled off from the skin after 24 hours. After 21 days of continuous administration, the major organs (heart, kidney, liver, lung, and spleen) of the rats were collected for H&E staining to assess the in vivo safety of the drug.

[0138] In vivo safety evaluation test results from Figure 16 As can be seen from the results, no significant histological damage was observed in any of the treatment groups compared to the control group, indicating that the administration of the aconitine inclusion complex microneedles did not induce significant systemic toxicity.

[0139] Obviously, the above embodiments of the present invention are merely examples for clearly illustrating the present invention, and are not intended to limit the implementation of the present invention. For those skilled in the art, other variations or modifications can be made based on the above description. It is impossible to exhaustively list all the implementation methods here. All obvious variations or modifications derived from the technical solutions of the present invention are still within the protection scope of the present invention.

Claims

1. A microneedle composition containing aconitine, characterized in that, The microneedle composition comprises, by mass percentage: 10-40% aconitine inclusion complex and 60-90% biocompatible polymer material; The biocompatible polymer material is selected from one or more of polyvinylpyrrolidone, polyvinyl alcohol, and methacrylamide gelatin; The aconitin inclusion complex is: 1) Aconitum carmichaelii, and 2) Cyclodextrins or cyclodextrin derivatives The resulting supramolecular inclusion complex; The cyclodextrin derivative is selected from at least one of sulfobutyl-β-cyclodextrin, sodium hydroxypropyl-β-cyclodextrin, and methyl-β-cyclodextrin.

2. The aconitine microneedle composition according to claim 1, characterized in that, The mass ratio of aconitine to cyclodextrin or cyclodextrin derivatives is 1:0.5 to 1:

4.

3. The aconitine microneedle composition according to claim 1, characterized in that, The mass ratio of aconitine to cyclodextrin or cyclodextrin derivatives is 1:1 to 1:

2.

4. The aconitine microneedle composition according to claim 1, characterized in that, The preparation of the aconitine inclusion complex includes the following steps: Aconitum carmichaelii and cyclodextrin or cyclodextrin derivatives were dissolved in water and stirred in a water bath at 30-50°C for 30-180 min to obtain a clear solution. The solution was then freeze-dried at -60-80°C to obtain the aconitine inclusion complex.

5. The aconitine microneedle composition according to claim 1, characterized in that, The mass ratio of the aconitine inclusion complex to the biocompatible polymer material is 1:2 to 1:

8.

6. A microneedle, characterized in that, The microneedles are prepared from raw materials comprising the aconitine microneedle composition as described in any one of claims 1-5 and water.

7. The method for preparing microneedles as described in claim 6, characterized in that, Includes the following steps: A biocompatible polymer material is dissolved in water to obtain a matrix solution; Add the aconitine inclusion complex to the matrix solution and stir until completely dissolved to obtain a microneedle solution; The microneedle solution is added dropwise into a mold, dried under negative pressure, to obtain the microneedles.

8. The preparation method according to claim 7, characterized in that, The biocompatible polymer material in the matrix solution has a mass percentage of 15-25%.

9. A microneedle patch comprising microneedles as described in claim 6 and a backing coupled to the microneedles.

10. The use of the aconitine microneedle composition according to any one of claims 1-5, or the microneedle according to claim 6, or the microneedle patch according to claim 9, in the preparation of a medicament for the treatment and / or prevention of rheumatoid arthritis, osteoarthritis, or local analgesia.

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