Dissoluble microneedle loaded with colchicine and iguratimod ethosome nano-drug and construction method of soluble microneedle loaded with colchicine and iguratimod ethosome nano-drug

Abstract

The invention provides a dissoluble microneedle loaded with colchicine and iguratimod ethosome nano-drugs and a construction method, the dissoluble microneedle comprises a needle body and a substrate, the substrate and the needle body both contain colchicine and iguratimod ethosome drugs, the dissoluble microneedle provided by the invention achieves the purpose of effectively treating gout, and the dissoluble microneedle has the advantages that the dissoluble microneedle can be used for treating gout; the traditional Chinese medicine composition can quickly relieve acute inflammation caused by gout diseases, has the effects of easing pain and resisting inflammation, repairs bone injury, and reduces gastrointestinal toxicity of colchicine and liver injury risk of iguratimod; according to the construction method of the dissoluble microneedle loaded with colchicine and iguratimod ethosome nano-drugs, the problems of drug degradation and ethosome agglomeration possibly caused by the difference of physicochemical properties of colchicine and iguratimod are solved fundamentally, the loss of drugs during mixing can be reduced, and the dispersity of ethosome can be optimized; the invention belongs to the technical field of preparation of nano-drugs, and is applied to preparation of drugs for treating gout.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of nanomedicine preparation, in particular to a soluble microneedle loaded with colchicine and etanercept liposome nanomedicine and a construction method thereof. BACKGROUND

[0002] Gout disease can cause acute inflammation of a single or multiple joints, commonly known as gout attack, which can cause joint redness, pain, and further damage to joint tissue. The aggravation of inflammatory response can cause cartilage destruction and bone erosion, and in extreme cases, it can evolve into permanent joint damage. In the clinical treatment of gout, the first choice of drug, colchicine, focuses on pain relief and anti-inflammatory in the acute phase, and cannot actively protect the skeleton. Etanercept is a small molecule chemical drug used for the treatment of autoimmune diseases, and is mainly used in clinical practice to improve the symptoms of rheumatoid arthritis and delay joint structure damage. The use of colchicine and etanercept at the same time can quickly relieve inflammation and repair bone damage. However, the current clinical dosage form of colchicine is colchicine tablets, and the specifications are mostly 0.5mg / tablet. When colchicine tablets are taken orally, patients are prone to ingest a toxic dose. The side effects of oral etanercept can damage liver cells, cause liver enzyme elevation, and also reduce blood platelets. In the prior art, there is a way of precise drug delivery using soluble microneedles. Colchicine and etanercept can be made into microneedles to reduce the toxicity of the two drugs. However, when colchicine and etanercept are made into soluble microneedles, the differences in the physicochemical properties of the two drugs can lead to incomplete drug dissolution and poor absorption. SUMMARY

[0003] The present application aims to provide one or more technical solutions to the existing problems, at least to provide a beneficial choice or create conditions.

[0004] To solve the above technical problems, the technical solutions adopted by the present application are as follows:

[0005] Firstly, the present application provides a construction method of a soluble microneedle loaded with colchicine and etanercept liposome nanomedicine, comprising the following steps:

[0006] S100, respectively constructing colchicine-loaded liposomes and etanercept-loaded liposomes;

[0007] S200, preparing a microneedle matrix solution;

[0008] S300, mixing the colchicine-loaded liposome concentrate and the etanercept-loaded liposome concentrate constructed in step S100 with the microneedle matrix solution prepared in step S200 according to a volume ratio of 1:1;

[0009] S400: Take the mixture of colchicine-loaded liposome and microneedle matrix prepared in step S300, dilute it 10 times with plasma water, add an equal volume of anhydrous ethanol, place it in an ultrasonic machine, and perform ultrasonic demulsification for 10 minutes.

[0010] S500: Take the mixture of the concentrated elamide liposome solution and microneedle matrix prepared in step S300, add plasma water to dilute it 10 times, then add an equal volume of anhydrous ethanol, place it in an ultrasonic machine, and perform ultrasonic demulsification for 10 minutes.

[0011] S600. Take equal volumes of the composite hydrogel containing colchicine alcohol body prepared in step S400 and the composite hydrogel containing aramod alcohol body prepared in step S500 into a 10×10 PDMS microneedle mold, place them in a vacuum drying oven for vacuum treatment, and keep them under a negative pressure of 0.08MPa for 20min.

[0012] After vacuum treatment, the microneedle mold is placed in a constant temperature and humidity chamber at a drying temperature of 30°C and a humidity of 55% for 6 to 8 hours.

[0013] S800, by gently peeling it off from the microneedle mold, can be obtained as colchicine- and aramod lyosome-soluble microneedles.

[0014] This method involves first preparing colchicine-loaded liposomes and elamodine-loaded liposomes separately, then combining them in equal volumes. Addressing the physicochemical properties of colchicine (slightly soluble in water) and elamodine (extremely poorly soluble in water), the liposome carrier significantly improves the solubility of both drugs. As a lipid nanocarrier, the liposome's alcohol content enhances the solubilization effect of poorly soluble drugs, solving the problems of incomplete drug dissolution and poor absorption in traditional formulations.

[0015] Colchicine and ellamod are encapsulated in liposomes. The lipid bilayer structure protects the drugs from external environmental factors (such as light, temperature, and pH), reduces drug degradation, minimizes losses during drug mixing, and optimizes liposome dispersibility. Utilizing soluble microneedles, the drugs can directly penetrate the stratum corneum, achieve effective drug loading, have minimal invasiveness, are easy to operate, and possess excellent biocompatibility. Through transdermal administration, the drugs effectively treat gout, rapidly relieve acute inflammation caused by gout, provide analgesic and anti-inflammatory effects, repair bone damage, and reduce the gastrointestinal toxicity of colchicine and the risk of liver damage associated with ellamod.

[0016] This method uses a 1:1 volume ratio to mix the drug-loaded liposomes and the microneedle matrix, ensuring initial uniform dispersion of the drug and matrix. Subsequent dilution and ultrasonic demulsification processes further break up the liposome aggregation, allowing colchicine and ellamod to be uniformly dispersed in the hydrogel matrix at the nanoscale. This avoids the problems of localized crystallization or uneven distribution of the drug in traditional mixing methods, ensuring consistent drug loading in each needle of the microneedle array and improving drug delivery stability.

[0017] As an extension of the above scheme, in step S100, the method for preparing colchicine-loaded liposomes is as follows:

[0018] Weigh out colchicine and egg yolk lecithin, dissolve them in anhydrous ethanol, and then perform water bath sonication under light-proof and sealed conditions at a power of 150W and a sonication temperature of 25℃. While sonicating, slowly inject pure water into the anhydrous ethanol solution. After completion, continue sonication for 15 minutes. Then, sonicate the resulting solution in an ice bath with a probe at a temperature of 4℃, a power of 125W, and a time of 5 minutes, with a 3-second sonication pause and a 2-second pause, to prepare a colchicine-loaded lyosome.

[0019] In this extended approach, by optimizing the solvent environment and using precise stepwise ultrasonic control, colchicine is efficiently encapsulated while maximizing the protection of drug stability and liposome structural integrity. The process is simple and controllable, providing a high-quality nanomedicine carrier for subsequent preparation of drug delivery systems such as drug-loaded microneedles.

[0020] As an extension of the above scheme, in step S100, the method for preparing the elamod liposome is as follows:

[0021] Aramod, egg yolk lecithin, and cholesterol were weighed and dissolved in anhydrous ethanol. Tween-80 and vitamin E were then added to the solution. The mixture was then stirred in a water bath at 50°C at 800 rpm while stirring. PBS buffer was slowly added dropwise while stirring. After the addition was complete, stirring was continued for 10 minutes. Subsequently, the mixture was sonicated in an ice bath with a 150W ultrasound probe for 30 seconds, followed by a 3-second sonication pause. This process prepared aramod propylforms. This extended method for preparing aramod propylforms addresses the low solubility of aramod and the need for optimized carrier stability. It achieves efficient drug loading and optimized carrier performance. Furthermore, it provides a performance-suitable nanomedicine carrier for subsequent integration with microneedle matrices and the construction of transdermal delivery systems. The process demonstrates strong practicality and reliability.

[0022] As an extension of the above scheme, in step S200, the preparation of the microneedle matrix solution involves: dissolving polyvinyl alcohol in pure water at 60°C with stirring, cooling to room temperature after complete dissolution, and then adding polyvinylpyrrolidone powder. This extended scheme solves the solubility and compatibility problem between polyvinylpyrrolidone and polyvinyl alcohol, ensuring matrix uniformity; it protects the subsequent mixing of the alcohol carrier and its own biocompatibility under "mild conditions," and ultimately, through the synergistic effect of polyvinylpyrrolidone and polyvinyl alcohol, endows the microneedle matrix with excellent formability, solubility, and biocompatibility, fully meeting the preparation requirements of "soluble microneedles loaded with alcohols," demonstrating extremely high process practicality and reliability.

[0023] As an extension of the above solution, the polyvinylpyrrolidone powder has a molecular weight of 58,000 and a polyvinyl alcohol viscosity of 12.0 mPa·s to 16.0 mPa·s. This extended solution endows the microneedles with the core characteristics of "penetration, easy dissolution, and minimal irritation," while ensuring the stability of the drug-loaded liposome and the long-term, stable release of the drug, thus meeting the therapeutic needs of colchicine and elamod liposome-soluble microneedles.

[0024] As an extension of the above scheme, in step S400, the concentration of colchicine in the prepared mixture is 1 mg / mL to 2 mg / mL. In this extended scheme, the viscosity of the mixture at this concentration combines the viscosity characteristics of polyvinyl alcohol and polyvinylpyrrolidone, making it suitable for filling and molding microneedles. It avoids the problem of excessive viscosity due to too high a concentration, which would prevent smooth filling of the mold, and also avoids the problem of insufficient drug loading or easy cracking after drying due to too low a concentration.

[0025] As an extension of the above scheme, in step S500, the concentration of elamod in the prepared mixture is 0.05 mg / mL to 0.5 mg / mL. As a drug used for chronic diseases such as rheumatoid arthritis, long-term use or local high-concentration accumulation of elamod may cause adverse reactions such as gastrointestinal irritation and abnormal liver function. In this extended scheme, the content of elamod does not disrupt the stability of the matrix, ensuring the molding effect and ensuring that the local drug concentration remains within the "effective therapeutic window." This avoids both insufficient efficacy due to excessively low concentrations and local tissue damage (such as synovial irritation and skin redness and swelling) caused by excessively high concentrations.

[0026] Based on the above scheme, the present invention also provides a soluble microneedle loaded with colchicine and ellamodolite nanomedicine, comprising a substrate and a needle body, wherein both the substrate and the needle body contain colchicine and ellamodolite.

[0027] This approach provides a soluble microneedle loaded with colchicine and elamod alcoholic nanomedicines. Both colchicine and elamod are loaded onto alcoholic bodies to create alcoholic nanomedicines, which are then further loaded into the soluble microneedles. Utilizing the advantages of these microneedles—direct penetration of the stratum corneum, efficient drug loading, minimal invasiveness, ease of operation, and excellent biocompatibility—this transdermal drug delivery method effectively treats gout, rapidly relieving acute inflammation caused by gout, providing analgesic and anti-inflammatory effects, repairing bone damage, and reducing the gastrointestinal toxicity of colchicine and the risk of liver damage from elamod. Loading colchicine and elamod onto alcoholic bodies improves drug loading efficiency, uniformity, and release uniformity. This approach is particularly suitable for chronic diseases requiring long-term anti-inflammatory and immunomodulatory treatments, significantly improving patient tolerance and compliance while enhancing efficacy.

[0028] As an extension of the above scheme, both the needle and the substrate are made of a mixed solution of a biocompatible soluble polymer material, colchicine olprosome nanomedicine, and ellamod olprosome nanomedicine. In this extended scheme, the two drugs are made into olprosomes before mixing, which solves the inherent defects of free drugs, improves drug stability, and reduces direct contact between the drugs and normal cells by encapsulating them in olprosomes. The colchicine and ellamod olprosome nanomedicines are mainly mixed with the polymer material, and their biological activity is well maintained.

[0029] As an extension of the above scheme, the biocompatible soluble polymeric materials are polyvinylpyrrolidone (PVP) and polyvinyl alcohol (PVA). Colchicine and ellamod liposomes are nanoscale carriers that easily aggregate or precipitate in the polymer matrix. By selecting PPV and PVA as biocompatible soluble polymeric materials, their complementary properties not only meet the core requirements of microneedles in terms of "formability, solubility, and drug loading," but also further improve the stability and safety of colchicine and ellamod liposome delivery.

[0030] This invention provides a soluble microneedle loaded with colchicine and elamodolite nanomedicine. The microneedle is made of materials with high biosafety and good biocompatibility, and is completely soluble in the body. While ensuring good mechanical properties, it also enables controlled release of the nanomedicine loaded on the microneedle, allowing for precise subcutaneous drug delivery, improving efficacy and reducing drug loss. The colchicine and elamodolite nanomedicine, as the main polymer materials, are mixed together, and their bioactivity is well maintained. Simultaneously, the solubility of the polymer materials facilitates the delivery of the nanomedicine to the subcutaneous layer, achieving orderly controlled release. The method for preparing the transdermal soluble microneedle of this invention is simple in process and allows for modification of the microneedle size and length, making it suitable for large-scale industrial production. This invention relates to the field of nanomedicine preparation technology and is applied to the preparation of soluble microneedles for treating gout. Attached Figure Description

[0031] The method of the present invention will be further described in detail below with reference to the accompanying drawings and embodiments.

[0032] Figure 1 This is a schematic diagram of the preparation process of soluble microneedles for the colchicine- and ellamod liposome nanomedicine of the present invention.

[0033] Figure 2 This is a schematic diagram of the soluble microneedle structure of the colchicine and ellamod liposome nanomedicine of the present invention.

[0034] Figure 3 The figures show the morphology of the microneedles in Comparative Example 1 of the present invention. A is a top view (partial) of Blank-MN; B is a side view (partial) of Blank-MN; C is a side view (partial) of Blank-MN; D is a 3D image (partial) of Blank-MN; and E is a scanning electron microscope image (partial) of Blank-MN.

[0035] Figure 4 The figures show the morphology of the microneedles in Embodiment 1 of the present invention. In the figures, E is a partial scanning electron microscope image; F is a partial top view; G is a partial oblique view; H is a partial side view; I is a partial 3D image; and J is a partial scanning electron microscope image.

[0036] Figure 5 The graph shows the mechanical performance test results of the microneedles in Comparative Example 1.

[0037] Figure 6 The graph shows the mechanical property test results of the microneedles in Example 1;

[0038] Figure 7The images show the mechanical properties of the microneedles before and after the test, with A being the image before the test and B being the image after the test.

[0039] Figure 8 Images are shown before and after the mechanical performance test of the microneedles in Example 1. In the image, C is the image before the test and D is the image after the test.

[0040] Figure 9 The above is a test image of the ex vivo skin puncture performance of Comparative Example 1 of the present invention. In the image, A is a methylene blue puncture image of Blank-MN and B is a coumarin 6 puncture image of Blank-MN.

[0041] Figure 10 The figure shows the ex vivo skin puncture performance test of Example 1 of the present invention. In the figure, C is the methylene blue puncture diagram of the microneedle in Example 1, and D is the coumarin 6 puncture diagram of the microneedle in Example 1.

[0042] Figure 11 This is a comparison chart of the dissolution performance of microneedles in Comparative Example 1 and Example 1 of the present invention. In the figure, A is the dissolution chart of microneedles in Comparative Example 1 at different time points, and B is the dissolution chart of microneedles in Example 1 at different time points.

[0043] Figure 12 The figures show a comparison of the skin irritation performance of microneedles in Embodiment 1 and Comparative Example 1 of the present invention. In the figures, A is the skin irritation performance evaluation chart of the microneedles provided in Comparative Example 1, and B is the skin irritation performance evaluation chart of the microneedles provided in Embodiment 1.

[0044] Figure 13 The graph shows the skin irritation and transdermal water loss of the microneedles in Embodiment 1 and Comparative Example 1 of the present invention. In the graph, A represents the transdermal water loss of normal skin, B represents the skin irritation and transdermal water loss of the microneedles in Comparative Example 1, and C represents the skin irritation and transdermal water loss of the microneedles in Embodiment 1.

[0045] Figure 14 The images show the changes in the appearance of the ankle joints in different groups after repeated administration of medication to treat gout in rats according to Example 1 of the present invention. In the images, a, b, c, d, e, f, and g represent different control groups. The left image in each control group represents the appearance of the rat's ankle joint in the initial state, and the right image represents the appearance of the rat's ankle joint after administration.

[0046] Figure 15 This is a statistical chart comparing the leg-raising time of each group with the baseline after repeated gout treatment in rats using microneedle intervention in Example 1 of the present invention. In the chart, each control group is represented by a bar, with the left bar representing the data before treatment and the right bar representing the data after treatment.

[0047] Figure 16This is a statistical chart comparing the bilateral paw pressure of rats after repeated gout treatment with microneedle intervention in Example 1 of the present invention with the baseline. The left bar represents the data before treatment, and the right bar represents the data after treatment.

[0048] Figure 17 These are imaging images of rats in different groups after repeated gout treatment with microneedle intervention in Example 1 of the present invention. In the images, a, b, c, d, e, f, and g represent different control groups.

[0049] Figure 18 This is a graph showing the serum TNF-α level of rats in different groups after repeated gout treatment with microneedle intervention in Example 1 of the present invention. In the graph, a, b, c, d, e, f, and g represent different control groups.

[0050] Figure 19 Example 1 of this invention: Serum IL-6 levels in rats after repeated gout treatment with microneedle intervention. In the figure, a, b, c, d, e, f, and g represent different control groups.

[0051] Figure 20 HE staining images of the ankle joints of rats in different groups after repeated gout treatment with microneedle intervention in Example 1 of this invention;

[0052] Figure 21 The images show safranin and fast green staining of the ankle joints of rats in different groups after repeated gout treatment with microneedle intervention in Example 1 of this invention.

[0053] Figure 22 This is a HE staining score of the ankle joints of rats in different groups after repeated gout treatment with microneedle intervention in Example 1 of the present invention.

[0054] Figure 23 This is a Safranin-Fix Green staining score chart of the ankle joints of rats in different groups after repeated gout treatment with microneedle intervention in Example 1 of the present invention.

[0055] Figure 24 This is a diagram showing the staining results of anti-tartrate acid phosphatase in rats after repeated gout treatment using microneedle intervention according to Example 1 of the present invention.

[0056] Figure 25 This is a statistical chart of osteoclasts in rats after repeated gout treatment using microneedle intervention in Example 1 of the present invention.

[0057] In the attached diagram: 100, the needle body of the microneedle; 200, the base of the microneedle. Detailed Implementation

[0058] This section will describe in detail specific embodiments of the present invention. Preferred embodiments of the present invention are shown in the accompanying drawings. The purpose of the drawings is to supplement the textual description with graphics, so that people can intuitively and vividly understand each technical feature and overall technical solution of the present invention, but they should not be construed as limiting the scope of protection of the present invention.

[0059] In the description of this invention, it should be understood that the orientation descriptions, such as up, down, front, back, left, right, etc., are based on the orientation or positional relationship shown in the accompanying drawings. They are only for the convenience of describing this invention and simplifying the description, and do not indicate or imply that the device or element referred to must have a specific orientation, or be constructed and operated in a specific orientation. Therefore, they should not be construed as limiting this invention.

[0060] In the description of this invention, if there are words such as "several", they mean one or more, "multiple" means two or more, "greater than", "less than", "exceeding" etc. are understood to exclude the number itself, and "above", "below", "within" etc. are understood to include the number itself.

[0061] In the description of this invention, unless otherwise explicitly defined, terms such as "set up," "install," and "connect" should be interpreted broadly, and those skilled in the art can reasonably determine the specific meaning of the above terms in this invention in conjunction with the specific content of the technical solution.

[0062] Reference Figures 1 to 25 Several embodiments of the present invention are given below.

[0063] like Figure 1 In some embodiments, a method for constructing soluble microneedles of colchicine and ellamod alcohol nanomedicine includes the following steps: S100, constructing colchicine-loaded alcohol bodies and ellamod alcohol bodies respectively.

[0064] S200, Preparation of microneedle matrix solution;

[0065] S300: The colchicine-loaded alcohol body concentrate and the ellamod-loaded alcohol body concentrate prepared in step S100 are mixed with the microneedle matrix solution prepared in step S200 at a volume ratio of 1:1.

[0066] S400: Take the mixture of colchicine-loaded liposome and microneedle matrix prepared in step S300, dilute it 10 times with plasma water, add an equal volume of anhydrous ethanol, place it in an ultrasonic machine, and perform ultrasonic demulsification for 10 minutes.

[0067] S500: Take the mixture of the concentrated elamide liposome solution and microneedle matrix prepared in step S300, add plasma water to dilute it 10 times, then add an equal volume of anhydrous ethanol, place it in an ultrasonic machine, and perform ultrasonic demulsification for 10 minutes.

[0068] S600: Take 200 μL of the composite hydrogel containing colchicine alcohol body prepared in step S400 and 200 μL of the composite hydrogel containing aramod alcohol body prepared in step S500 into a 10×10 PDMS microneedle mold, place it in a vacuum drying oven for vacuum treatment, and keep it under a negative pressure of 0.08 MPa for 20 min.

[0069] After vacuum treatment, the microneedle mold is placed in a constant temperature and humidity chamber at a drying temperature of 30°C and a humidity of 55% for 6 to 8 hours.

[0070] S800, by gently peeling it off from the microneedle mold, can be obtained as colchicine- and aramod lyosome-soluble microneedles.

[0071] The construction method provided in this embodiment involves first preparing colchicine-loaded liposomes and elamodine-loaded liposomes separately, and then combining them in equal volumes. Addressing the physicochemical properties of colchicine ("slightly soluble in water") and elamodine ("extremely poorly soluble in water"), the liposome carrier significantly improves the solubility of both drugs. As a lipid nanocarrier, the liposome's alcohol content enhances the solubilization effect of poorly soluble drugs, solving the problems of incomplete drug dissolution and poor absorption in traditional formulations.

[0072] Colchicine and ellamod are encapsulated in liposomes. The lipid bilayer structure protects the drugs from external environmental factors (such as light, temperature, and pH), reduces drug degradation, minimizes losses during drug mixing, and optimizes liposome dispersibility. Utilizing soluble microneedles, the drugs can directly penetrate the stratum corneum, achieve effective drug loading, have minimal invasiveness, are easy to operate, and possess excellent biocompatibility. Through transdermal administration, the drugs effectively treat gout, rapidly relieve acute inflammation caused by gout, provide analgesic and anti-inflammatory effects, repair bone damage, and reduce the gastrointestinal toxicity of colchicine and the risk of liver damage associated with ellamod.

[0073] This method uses a 1:1 volume ratio to mix the drug-loaded ethanol body with the microneedle matrix, ensuring initial uniform dispersion of the drug and matrix. Subsequent dilution and ultrasonic demulsification further break up ethanol body aggregation, allowing colchicine and ellamod to be uniformly dispersed in the hydrogel matrix at the nanoscale. This avoids the problems of localized crystallization or uneven distribution of the drug in traditional mixing methods, ensuring consistent drug loading in each needle of the microneedle array, improving drug delivery stability, and reducing irritation caused by excessively high local drug concentrations. Dilution and ethanol adjustment optimize the system viscosity, making it suitable for subsequent filling and molding of the PDMS mold. At the same time, the addition of ethanol helps inhibit microbial growth and improves the storage stability of the formulation. The preparation of soluble microneedles ensures that the microneedles are fully solidified (maintaining the sharpness of the needle tip for easy skin penetration) while avoiding high temperatures that could damage the liposome structure and drug activity. At the same time, appropriate humidity can prevent the microneedles from becoming brittle due to excessive dryness. This allows the final soluble microneedles to gradually dissolve upon contact with skin tissue fluid, releasing the liposomes that encapsulate the drug. There is no need to remove the microneedles afterward, reducing the risk of skin irritation and infection. Furthermore, the water-soluble matrix (such as hyaluronic acid) itself has the characteristics of moisturizing and good biocompatibility, further improving the safety of drug use.

[0074] In this embodiment, the method for preparing colchicine-loaded liposomes in step S100 is as follows:

[0075] Colchicine and egg yolk lecithin were weighed and dissolved in anhydrous ethanol. Then, under light-protected and sealed conditions, a water bath sonication was performed at a power of 150W and a temperature of 25℃. While sonicating, pure water was slowly injected into the anhydrous ethanol solution. After sonication, the solution was sonicated for another 15 minutes. The resulting solution was then sonicated in an ice bath at a temperature of 4℃ and a power of 125W for 5 minutes, with a 3-second sonication pause followed by a 2-second pause. This process prepared colchicine-loaded lecithin. Egg yolk lecithin was used as the membrane material for the lecithin (it is rich in unsaturated fatty acids and readily forms a stable phospholipid bilayer). It was dissolved in anhydrous ethanol along with colchicine. The solubilizing effect of ethanol on phospholipids (lowering the critical micelle concentration of phospholipids) laid the foundation for subsequent vesicle formation. Simultaneously, the strong polarity of anhydrous ethanol could initially improve the solubility of colchicine (which is highly lipid-soluble), preventing drug crystallization.

[0076] By optimizing the solvent environment and using precise stepwise ultrasonic control, colchicine was efficiently encapsulated while maximizing the protection of drug stability and liposome structural integrity. The process is simple and controllable, providing a high-quality nanomedicine carrier for subsequent preparation of drug delivery systems such as drug-loaded microneedles.

[0077] In step S100, the method for preparing elamod liposomes is as follows:

[0078] Weigh out elamod, egg yolk lecithin, and cholesterol, dissolve them in anhydrous ethanol, then add Tween-80 and vitamin E to the solution. The solution is then stirred in a water bath at 50°C at 800 rpm while stirring. PBS buffer is slowly added dropwise while stirring. After the addition is complete, stirring continues for 10 minutes. Subsequently, the solution is sonicated in an ice bath for 30 seconds with a 3-second sonication pause, to prepare elamod liposomes. Lecithin, as the main film-forming material, uses its amphiphilic structure to encapsulate elamod (which is highly lipophilic) through hydrophobic interactions. The addition of cholesterol allows it to embed into the phospholipid bilayer, reducing membrane fluidity, decreasing the probability of vesicle fusion, and significantly improving the physical stability of the liposomes. Tween-80 (a nonionic surfactant) increases the solubility of elamod in the ethanol-water mixture through solubilization, while also reducing interfacial tension, promoting uniform dispersion of phospholipid vesicles, and reducing drug crystallization. Vitamin E, as an antioxidant, can inhibit the degradation of elamod (containing amide bonds and easily oxidized) during the preparation process. Simultaneously, its lipid solubility enhances compatibility with phospholipid membranes, further stabilizing the carrier structure. This process for preparing elamod-loaded propyl bodies addresses the drug's low solubility and the need for optimized carrier stability. It achieves efficient drug loading and optimized carrier performance, and provides a performance-suitable nanomedicine carrier for subsequent integration with microneedle matrices to construct transdermal delivery systems. The process demonstrates strong practicality and reliability.

[0079] In step S200, the preparation of the microneedle matrix solution involves dissolving polyvinyl alcohol in pure water at 60°C with stirring. After complete dissolution, the solution is cooled to room temperature, and then polyvinylpyrrolidone powder is added. This solves the solubility and compatibility problem between polyvinylpyrrolidone and polyvinyl alcohol, ensuring matrix homogeneity. The "mild conditions" protect the subsequent mixing of the alcohol carrier and the microneedle matrix itself from biocompatibility issues. Ultimately, through the synergistic effect of polyvinylpyrrolidone and polyvinyl alcohol, the microneedle matrix is ​​endowed with excellent formability, solubility, and biocompatibility, perfectly meeting the preparation requirements of "soluble microneedles loaded with alcohols." The process is highly practical and reliable. The polyvinylpyrrolidone powder has a molecular weight of 58,000, and the polyvinyl alcohol viscosity is between 12.0 mPa·s and 16.0 mPa·s. This ensures that the prepared microneedles possess the core characteristics of "penetration, easy dissolution, and minimal irritation," while also ensuring the stability of the drug-loaded alcohol and the long-term, stable release of the drug, meeting the therapeutic needs of soluble microneedles loaded with colchicine and elamod alcohols.

[0080] In step S400, the concentration of colchicine in the prepared mixture is 1 mg / mL to 2 mg / mL. In this extended scheme, the viscosity of the mixture at this concentration combines the viscosity characteristics of polyvinyl alcohol and polyvinylpyrrolidone, making it suitable for filling and molding microneedles. It avoids the problem of excessive viscosity due to too high a concentration, which would prevent smooth filling of the mold, and also avoids the problem of insufficient drug loading or easy cracking after drying due to too low a concentration.

[0081] In step S500, the concentration of elamod in the prepared mixture is 0.05 mg / mL to 0.5 mg / mL. As a drug used for chronic diseases such as rheumatoid arthritis, long-term use or local high-concentration accumulation of elamod may cause adverse reactions such as gastrointestinal irritation and abnormal liver function. In this extended formulation, the content of elamod does not compromise matrix stability, ensuring the molding effect and guaranteeing that the local drug concentration remains within the "effective therapeutic window." This avoids both insufficient efficacy due to excessively low concentrations and local tissue damage (such as synovial irritation and skin redness and swelling) caused by excessively high concentrations.

[0082] In some embodiments, the present invention also provides a soluble microneedle loaded with colchicine and elamodolite nanomedicine, which is prepared by the method for constructing soluble microneedles loaded with colchicine and elamodolite nanomedicine as described in any one or more of the above embodiments, comprising a needle body 100 and a substrate 200, wherein both the substrate and the needle body contain colchicine and elamodolite.

[0083] like Figure 2 As shown, in this embodiment, a soluble microneedle loaded with colchicine and elamod alcoholic nanomedicine is provided. Both colchicine and elamod are loaded onto alcoholic bodies to form alcoholic nanomedicines. These nanomedicines are then further loaded into the soluble microneedles. Utilizing the advantages of soluble microneedles—direct penetration of the stratum corneum, effective drug loading, minimal invasiveness, ease of operation, and excellent biocompatibility—transdermal drug delivery effectively treats gout, rapidly relieving acute inflammation caused by gout, providing analgesic and anti-inflammatory effects, repairing bone damage, and reducing the gastrointestinal toxicity of colchicine and the risk of liver damage from elamod. Loading colchicine and elamod onto alcoholic bodies improves drug loading efficiency, uniformity, and release uniformity, making it particularly suitable for chronic diseases requiring long-term anti-inflammatory and immunomodulatory effects. This significantly improves patient tolerance and compliance while enhancing efficacy.

[0084] In this embodiment, the microneedle body is 850 μm long, the microneedle array is 10×10, the distance between adjacent needle tips is 750 μm, the size of the microneedle base is 350 μm×350 μm, the colchicine content is 0.3 mg, the elamod content is 0.02 mg, and the ratio of the two drugs is approximately 15:1. This ratio reflects both the dominant role of colchicine in rapid anti-inflammatory action and the synergistic effect of long-term immune regulation achieved through low-dose elamod.

[0085] In some optional embodiments, both the needle and the substrate are made from a mixed solution of a biocompatible soluble polymer material, colchicine olprosome nanomedicine, and ellamod olprosome nanomedicine. Preparing the two drugs as olprosomes before mixing overcomes the inherent defects of free drugs, improves drug stability, and reduces direct contact between the drug and normal cells. The colchicine and ellamod olprosome nanomedicines are primarily preserved through simple mixing with the polymer material, thus maintaining their bioactivity effectively.

[0086] In this embodiment, the biocompatible soluble polymeric materials are polyvinylpyrrolidone (PVP) and polyvinyl alcohol (PVA). Colchicine lyosomes and ellamod lyosomes are nanoscale carriers that easily aggregate or precipitate in a polymer matrix. PVP and PVA are selected as biocompatible soluble polymeric materials because their complementary properties not only meet the core requirements of microneedles in terms of "formability, solubility, and drug loading," but also further improve the stability and safety of colchicine and ellamod lyosome delivery.

[0087] The following examples further illustrate the present invention in detail. It should also be understood that the following examples are only for further explanation of the present invention and should not be construed as limiting the scope of protection of the present invention. Any non-essential improvements and adjustments made by those skilled in the art based on the above description of the present invention are within the scope of protection of the present invention. The specific process parameters, etc., in the following examples are merely examples within a suitable range; that is, those skilled in the art can make appropriate selections within the appropriate range based on the description herein, and are not intended to be limited to the specific values ​​in the examples below.

[0088] Example 1: A method for constructing soluble microneedles of colchicine and ellamod liposome nanomedicine:

[0089] A. Construction of colchicine- and ellamod nanoparticles:

[0090] The colchicine hydroxyl body was constructed as follows: 20 mg of colchicine and 200 mg of egg yolk lecithin were weighed and dissolved in 5.06 mL of anhydrous ethanol. Then, under light-protected and sealed conditions, the solution was subjected to a water bath sonication at 150 W and 25 °C. While sonicating, 5.78 mL of pure water was slowly injected into the anhydrous ethanol solution. After sonication, the solution was continued for 15 min. Subsequently, the solution was subjected to a probe-ice bath sonication at 4 °C and 125 W for 5 min, with a 3-second sonication pause followed by a 2-second pause. The colchicine hydroxyl body was then prepared. The elamod hydroxyl body was constructed as follows: 5 mg of elamod, 50 mg of egg yolk lecithin, and 10 mg of cholesterol were weighed and dissolved in 5 mL of anhydrous ethanol. Then, 100 mg of Tween-80 and 20 mg of vitamin E were added to the solution. The solution was then stirred in a water bath at 50 °C at a stirring speed of 800 r / min. While stirring, 5 mL of pure water was slowly added dropwise. After adding PBS buffer, continue stirring for 10 minutes, then perform ice bath sonication under a 150W sonic probe for 30 seconds, with a 3-second sonication pause and a 2-second pause, to prepare aramod lyosomes.

[0091] B. Construction of soluble microneedles loaded with colchicine and elamodol: A 10×10 PDMS microneedle mold was used to prepare soluble microneedles by vacuum method;

[0092] Preparation of microneedle matrix solution: Dissolve 0.7g of polyvinyl alcohol in 10mL of pure water at 60℃ and stir. After complete dissolution, cool to room temperature and then add 5g of polyvinylpyrrolidone to prepare the solution.

[0093] The lyophilized colchicine alcohol body and ellamod alcohol body concentrate were mixed with the microneedle matrix solution at a volume ratio of 1:1. Two units of matrix solution were taken first, and then equal amounts of colchicine alcohol body and ellamod alcohol body concentrate were taken and poured into the two units of matrix solution respectively to obtain the colchicine alcohol body and microneedle matrix mixture and the ellamod alcohol body and microneedle matrix mixture.

[0094] First, 200 μL of a mixture of colchicine propyl ester and microneedle matrix was precisely pipetted, diluted 10-fold with plasma water, and then an equal volume of anhydrous ethanol was added. The mixture was then placed in an ultrasonic machine for 10 minutes for ultrasonic demulsification. The colchicine concentration in the mixture was measured using a liquid chromatography instrument. The calculated colchicine content in 200 μL was approximately 1.5 mg / mL. Next, 200 μL of a mixture of elastolide propyl ester concentrate and microneedle matrix was precisely pipetted, diluted 10-fold with plasma water, and then an equal volume of anhydrous ethanol was added. The mixture was then placed in an ultrasonic machine for 10 minutes for ultrasonic demulsification. The elastolide concentration in the mixture was measured using a UV spectrophotometer. The calculated elastolide concentration in 200 μL was approximately 0.1 mg / mL.

[0095] 200 μL of the composite hydrogel containing colchicine alcohol and ellamod alcohol was precisely pipetted into a microneedle mold and placed in a vacuum drying oven for vacuum treatment at a negative pressure of 0.08 MPa for 20 min.

[0096] After vacuum treatment, the microneedle mold is placed in a constant temperature and humidity chamber to dry for 6-8 hours;

[0097] Soluble microneedles loaded with colchicine and aramod liposomes can be obtained by gently peeling them off from the microneedle mold.

[0098] Comparative Example 1:

[0099] A method for constructing soluble microneedles, comprising the following steps:

[0100] This study used a 10×10 PDMS microneedle mold and employed a vacuum method to prepare soluble microneedles. 0.35 g of PVA powder was heated and dissolved in 10 mL of deionized water to obtain a 3.5 wt% PVA solution. After cooling to room temperature, 2.5 g of PVP powder was added, and the solution was allowed to stand at room temperature until all the PVP powder dissolved, yielding a composite hydrogel solution. 200 μL of the composite hydrogel was precisely pipetted into the microneedle mold and placed in a vacuum drying oven for vacuum treatment at 0.08 MPa for 20 min. After vacuum treatment, the microneedle mold was placed in a constant temperature and humidity chamber for drying for 6-8 h. Demolding: The soluble microneedles, abbreviated as Blank-MN, were obtained by gently peeling them off from the microneedle mold.

[0101] The difference from Example 1 is that the soluble microneedles provided in Comparative Example 1 do not contain liposome nanomedicine in the needle body and substrate, while the other parameters and operations are basically the same as in Example 1.

[0102] Depend on Figure 3 , Figure 4 The morphology of the microneedles prepared in Example 1 and Comparative Example 1 can be seen.

[0103] Depend on Figure 5 , Figure 6 , Figure 7 , Figure 8 As can be seen, the mechanical properties of the microneedles prepared in Example 1 and Comparative Example 1 were measured. According to the stress-displacement curves, the stress of the soluble microneedles prepared in Comparative Example 1 reached 7.5 N, and the stress of the soluble microneedles loaded with colchicine and ellamod nanomedicine prepared in Example 1 also reached 7.5 N. Furthermore, digital photographs of the microneedles after mechanical strength testing showed that only significant bending occurred at the tip of the microneedle, without any tip breakage, indicating that the prepared microneedles exhibited excellent flexibility.

[0104] Depend on Figure 9 , Figure 10 As can be seen, the ex vivo skin penetration performance of the microneedles prepared in Example 1 and Comparative Example 1 was measured. The experimental results show that the soluble microneedles prepared in Example 1 and Comparative Example 1 can easily penetrate the skin.

[0105] Depend on Figure 11 As can be seen, the solubility of the microneedles prepared in Example 1 and Comparative Example 1 was evaluated. The experimental results indicate that Example 1 and Comparative Example 1 have good solubility, and the needle body of the microneedles can be basically completely dissolved after 20 minutes.

[0106] Depend on Figure 12 As can be seen, the skin irritation of the microneedles prepared in Example 1 and Comparative Example 1 was evaluated. Within 1 hour of removing the microneedles, the skin recovered completely, suggesting that both microneedles have low irritation to rat skin.

[0107] Depend on Figure 13 As can be seen, the skin irritation assessment of the microneedles prepared in Example 1 and Comparative Example 1 showed that the transdermal water loss returned to normal after 2 hours, suggesting that the microneedles only pierce the stratum corneum and do not damage the dermis.

[0108] Depend on Figure 14 , Figure 15 , Figure 16 , Figure 17 It is evident that the microneedles prepared in Example 1 have excellent analgesic and anti-inflammatory effects.

[0109] Depend on Figure 18 , Figure 19 , Figure 20 , Figure 21 , Figure 22 , Figure 23 It is evident that the microneedles prepared in Example 1 have excellent analgesic and anti-inflammatory effects.

[0110] Depend on Figure 24 , Figure 25It is evident that the microneedles prepared in Example 1 have excellent analgesic, anti-inflammatory, and bone-protective effects.

[0111] Based on the above description, during the preparation of soluble microneedles loaded with colchicine and elamod liposome nanomedicine, the differences in the physicochemical properties of colchicine and elamod, such as solubility, lipid solubility, and tolerance to ultrasound / solvents, may lead to problems such as drug degradation, liposome aggregation, uneven drug distribution, poor needle tip morphology, insufficient mechanical strength of microneedles, and abnormal release behavior. This invention first prepares two drugs into liposomes to enhance drug solubility and permeability, and then loads the liposomes onto soluble microneedles to improve their solubility and stability, while reducing drug irritation to the skin and further enhancing drug delivery efficiency. Stepwise preparation and ultrasonic demulsification ensure drug loading stability, and vacuum drying combined with constant temperature and humidity curing ensures the morphology and solubility of the microneedles. Ultrasonic demulsification evenly disperses the liposomes and matrix, preventing drug aggregation and ensuring uniform drug distribution within the microneedles, reducing irritation caused by excessively high local drug concentrations. Dilution and ethanol adjustment optimize system viscosity, adapting to subsequent PDMS mold filling and molding. The addition of ethanol also helps inhibit microbial growth and improves the storage stability of the formulation. This allows the prepared soluble microneedles loaded with colchicine and elamod liposome nanomedicines to rapidly relieve acute inflammation caused by gout, achieving analgesic and anti-inflammatory effects, repairing bone damage, and reducing the gastrointestinal toxicity of colchicine and the risk of liver damage from elamod.

[0112] The preferred embodiments of the present invention have been described in detail above, but the present invention is not limited to the embodiments described. Those skilled in the art can make various equivalent modifications or substitutions without departing from the spirit of the present invention, and these equivalent modifications or substitutions are all included within the scope defined by the claims of this application.

Claims

1. A method for constructing soluble microneedles loaded with colchicine and ellamodol liposome nanomedicine, characterized in that, Includes the following steps: S100 was used to construct colchicine-loaded ologens and ellamod-loaded ologens, respectively. S200, for preparing microneedle matrix solution; S300, the colchicine-loaded liposome concentrate and the ellamod-loaded liposome concentrate prepared in step S100 are mixed with the microneedle matrix solution prepared in step S200 at a volume ratio of 1:

1. S400: Take the mixture of colchicine-loaded liposome and microneedle matrix prepared in step S300, dilute it 10 times with plasma water, add an equal volume of anhydrous ethanol, place it in an ultrasonic machine, and perform ultrasonic demulsification for 10 minutes. S500, take the mixture of the concentrated aramod liposome solution and microneedle matrix prepared in step S300, add plasma water to dilute it 10 times, then add an equal volume of anhydrous ethanol, place it in an ultrasonic machine, and perform ultrasonic demulsification for 10 minutes. S600, take equal volumes of the composite hydrogel containing colchicine alcohol body prepared in step S400 and the composite hydrogel containing aramod alcohol body prepared in step S500 into a PDMS microneedle mold, place them in a vacuum drying oven for vacuum treatment, and keep them under a negative pressure of 0.08MPa for 20min. After vacuum treatment, the S700 microneedle mold is placed in a constant temperature and humidity chamber at a drying temperature of 30℃ and a humidity of 55% for 6 to 8 hours. S800 can be easily peeled off from the microneedle mold to obtain colchicine- and aramod lyosome-soluble microneedles.

2. The method for constructing soluble microneedles of colchicine and ellamodol liposome nanomedicine according to claim 1, characterized in that: In step S100, the method for preparing colchicine-loaded liposomes is as follows: Weigh out colchicine and egg yolk lecithin, dissolve them in anhydrous ethanol, and then perform water bath sonication under light-proof and sealed conditions at a power of 150W and a sonication temperature of 25℃. While sonicating, slowly inject pure water into the anhydrous ethanol solution. After completion, continue sonication for 15 minutes. Then, sonicate the resulting solution in an ice bath with a probe at a temperature of 4℃, a power of 125W, and a time of 5 minutes, with a 3-second sonication pause and a 2-second pause, to prepare a colchicine-loaded lyosome.

3. The method for constructing soluble microneedles of colchicine and ellamodol liposome nanomedicine according to claim 1, characterized in that: In step S100, the method for preparing elamod liposomes is as follows: Weigh out elamod, egg yolk lecithin and cholesterol, dissolve them in anhydrous ethanol, then add Tween-80 and vitamin E to the solution, and then stir in a water bath at 50°C at a stirring speed of 800 r / min. While stirring, slowly add PBS buffer to the solution. After the addition is complete, continue stirring for 10 min. Then, sonicate in an ice bath with a 150W sonication probe for 30 s, with a 3 s sonication pause and a 2 s pause, to prepare elamod ol corpuscles.

4. The method for constructing soluble microneedles of colchicine and ellamodol liposome nanomedicine according to claim 1, characterized in that: In step S200, the method for preparing the microneedle matrix solution is as follows: polyvinyl alcohol is dissolved in pure water at 60°C and stirred. After complete dissolution, it is cooled to room temperature, and then polyvinylpyrrolidone powder is added.

5. The method for constructing soluble microneedles of colchicine and ellamodol liposome nanomedicine according to claim 4, characterized in that: The polyvinylpyrrolidone powder has a molecular weight of 56,000 to 60,000 and a polyvinyl alcohol viscosity of 12.0 mPa·s to 16.0 mPa·s.

6. The method for constructing soluble microneedles of colchicine and ellamodol liposome nanomedicine according to claim 1, characterized in that: In step S400, the concentration of colchicine in the prepared mixture is 1 mg / mL to 2 mg / mL.

7. The method for constructing soluble microneedles of colchicine and ellamodol liposome nanomedicine according to claim 1, characterized in that: In step S500, the concentration of ellamod in the prepared mixture is from 0.05 mg / mL to 0.5 mg / mL.

8. A soluble microneedle loaded with colchicine and ellamodol liposome nanomedicine, characterized in that: The soluble microneedles, prepared by the method of constructing colchicine and ellamodolite nanomedicine according to any one of claims 1 to 7, include a needle body (100) and a substrate (200), wherein both the substrate (200) and the needle body (100) contain colchicine and ellamodolite.

9. The soluble microneedles of colchicine and ellamodol liposome nanomedicine according to claim 8, characterized in that: Both the needle (100) and the substrate (200) are made of a mixed solution of a biocompatible soluble polymer material, colchicine alcohol nanoparticles, and ellamod alcohol nanoparticles.

10. The soluble microneedles of colchicine and ellamodol liposome nanomedicine according to claim 9, characterized in that: The biocompatible soluble polymeric materials mentioned are polyvinylpyrrolidone and polyvinyl alcohol.

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