Jinhuang powder drug-loaded microneedle for improving gouty arthritis and preparation method of Jinhuang powder drug-loaded microneedle

Abstract

The invention relates to a Jinhuang powder drug-loaded microneedle for improving gouty arthritis and a preparation method thereof. The preparation method comprises the following steps: step 1, researching the action mechanism of the Jinhuang powder for treating the gouty arthritis; 2, according to the research result, increasing the proportion of cortex phellodendri, cortex magnoliae officinalis and liquorice in the original golden-huang powder prescription, and reducing the proportion of other medicinal materials to obtain a new golden-huang powder prescription; and step 3, preparing the Jinhuang powder drug-loaded microneedle on the basis of the new prescription of the Jinhuang powder. A golden yellow powder medicinal material is loaded through a microneedle, painless drug delivery of the microneedle is utilized, the delivery efficiency is higher than that of traditional transdermal drug delivery, good biocompatibility is achieved for delivery of protein and nucleic acid macromolecular drugs, the drug delivery efficiency can be improved, the drugs are delivered to specific parts in a targeted mode, adverse reactions of the drugs are reduced, and the utilization rate of the drugs is increased; the proportion of the golden cypress, the mangnolia officinalis and the liquorice in an original golden-yellow powder prescription is increased, and the treatment effect of the golden-yellow powder drug-loaded microelement on gouty arthritis is further enhanced.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field, in particular to a kind of gold yellow powder drug-loaded microneedle for improving gout arthritis and preparation method thereof. BACKGROUND

[0002] The drug for treating gout arthritis is mostly oral administration, which is easy to produce adverse drug reactions, leading to nausea and vomiting in patients, and the absorption and utilization rate of the drug is low. SUMMARY

[0003] The present application aims to provide a kind of gold yellow powder drug-loaded microneedle for improving gout arthritis, to solve the above problems existing in the prior art.

[0004] The technical scheme for solving the above technical problems is as follows: A preparation method of a gold yellow powder drug-loaded microneedle for improving gout arthritis, comprising the following steps: Step 1, study the mechanism of action of gold yellow powder in treating gout arthritis; Step 2, according to the research results, increase the proportion of three medicinal materials of Huangbai, Hupo and Gancao in the original gold yellow powder prescription, and reduce the proportion of other medicinal materials, to obtain a new gold yellow powder prescription; Step 3, prepare gold yellow powder drug-loaded microneedle based on the new gold yellow powder prescription.

[0005] The present application has the following beneficial effects: by loading gold yellow powder medicinal materials on microneedles, using the painless drug delivery of microneedles, higher delivery efficiency than traditional transdermal drug delivery, and good biocompatibility for protein and nucleic acid macromolecular drug delivery, the drug delivery efficiency can be improved, and the drug can be targeted to deliver to a specific site, reducing adverse drug reactions and improving drug utilization; and by studying the mechanism of action of gold yellow powder in treating gout arthritis, increasing the proportion of three medicinal materials of Huangbai, Hupo and Gancao in the original gold yellow powder prescription, and further enhancing the therapeutic effect of gold yellow powder drug-loaded microneedle on gout arthritis.

[0006] On the basis of the above technical scheme, the present application can also be improved as follows.

[0007] Further, step 1, study the mechanism of action of gold yellow powder in treating gout arthritis, specifically includes: Step 1.1, search for related targets of gold yellow powder in treating gout arthritis; Step 1.2, perform "medicinal material-target-disease" target analysis; Step 1.3, find the signal pathways in which the differential genes are involved through KEGG (Kyoto Encyclopedia of Genes and Genomes) enrichment analysis; Step 1.4, reveal the molecular mechanism of gold yellow powder in treating gout arthritis by analyzing the distribution of differential genes in KEGG pathways; Step 1.5, in combination with KEGG pathway enrichment analysis, it is determined that the Jinhuangsan can treat gouty arthritis through two pathways of TLR4 and NF-kB; Step 1.6, molecular docking of each component of Jinhuangsan and the two pathways of TLR4 and NF-kB is carried out, and finally it is determined that quercetin, beta-sitosterol and stigmasterol can directly act on the two pathways of TLR4 and NF-kB; Step 1.7, it is determined that quercetin, beta-sitosterol and stigmasterol are distributed in three kinds of medicinal materials of Phellodendri Chinensis Cortex, Magnoliae Officinalis Cortex and Glycyrrhizae Radix.

[0008] The further beneficial effect of the above is that quercetin, beta-sitosterol and stigmasterol can respectively dock the two pathways of TLR4 and NF-kB to treat gouty arthritis, and it is determined that quercetin, beta-sitosterol and stigmasterol are distributed in three kinds of medicinal materials of Phellodendri Chinensis Cortex, Magnoliae Officinalis Cortex and Glycyrrhizae Radix, which provides support for subsequent modification of the original Jinhuangsan prescription.

[0009] Further, step 3, the Jinhuangsan drug-loaded microneedle prepared on the basis of the new Jinhuangsan prescription specifically comprises: Step 3.1, decocting the medicinal materials in the new Jinhuangsan prescription to obtain Jinhuangsan freeze-dried powder; Step 3.2, preparing Jinhuangsan drug-loaded microneedle from Jinhuangsan freeze-dried powder.

[0010] Further, step 3.1, decocting the medicinal materials in the new Jinhuangsan prescription to obtain Jinhuangsan freeze-dried powder specifically comprises: adding water to the medicinal materials in the new Jinhuangsan prescription at a ratio of 1:9 for decoction, then centrifuging the obtained medicinal liquid through a centrifuge and fine filtering to obtain a preliminary extract, placing the preliminary extract in a centrifuge tube, centrifuging and fine filtering, and finally oscillating to obtain a final extract, pre-freezing the final extract for a period of time, and drying to obtain Jinhuangsan freeze-dried powder. The further beneficial effect of the above is that the purified Jinhuangsan freeze-dried powder has less impurities and is convenient for subsequent preparation of Jinhuangsan drug-loaded microneedle.

[0011] Further, it further comprises step 4, morphology and physical property detection of Jinhuangsan drug-loaded microneedle.

[0012] The further beneficial effect of the above is that the Jinhuangsan drug-loaded microneedle prepared by different magnifications of optical electron microscope has stable morphology; the physical property of the Jinhuangsan drug-loaded microneedle does not change adversely compared with non-drug-loaded microneedle, and the transdermal drug delivery standard is reached.

[0013] Another technical scheme of the application is as follows: A Jinhuangsan drug-loaded microneedle for improving gouty arthritis is prepared by the preparation method of the Jinhuangsan drug-loaded microneedle for improving gouty arthritis as described above. BRIEF DESCRIPTION OF DRAWINGS

[0014] Figure 1 This is a flowchart illustrating the preparation method of a golden powder-loaded microneedle for improving gouty arthritis according to the present invention. Figure 2 This is a database diagram of a method for preparing a golden powder-loaded microneedle for improving gouty arthritis according to the present invention. Figure 3 This is a target analysis diagram of "medicinal material-target point-disease" for the preparation method of the golden powder-loaded microneedles for improving gouty arthritis according to the present invention. Figure 4 Bubble diagram of KEGG pathway enrichment analysis for a method of preparing a golden powder-loaded microneedle for improving gouty arthritis according to the present invention. Figure 5 This is a molecular docking result diagram of the preparation method of the golden powder drug-loaded microneedles for improving gouty arthritis according to the present invention. Figure 6 This is a structural diagram of a golden powder-loaded microneedle for improving gouty arthritis according to the present invention; Figure 7 This is a scanning electron microscope image of a golden powder-loaded microneedle for improving gouty arthritis according to the present invention. Detailed Implementation

[0015] The principles and features of the present invention are described below with reference to the accompanying drawings. The examples given are only for explaining the present invention and are not intended to limit the scope of the present invention.

[0016] Example 1 like Figures 1 to 7 As shown, a method for preparing a golden powder-loaded microneedle for improving gouty arthritis includes the following steps: Step 1: Study the mechanism of action of Jinhuang San in treating gouty arthritis; Step 2: Based on the research results, increase the proportion of three medicinal materials, namely Phellodendron bark, Magnolia bark, and Licorice, in the original Jinhuang San formula, and decrease the proportion of other medicinal materials to obtain the new Jinhuang San formula; Step 3: Prepare Jinhuangsan drug-loaded microneedles based on the new Jinhuangsan formula.

[0017] By loading Jin Huang San (a traditional Chinese medicine) onto microneedles, the painless drug delivery, higher delivery efficiency than traditional transdermal drug delivery, and good biocompatibility for delivering protein and nucleic acid macromolecules can be utilized to improve drug delivery efficiency, target drugs to specific sites, reduce adverse drug reactions, and improve drug utilization. Furthermore, by studying the mechanism of action of Jin Huang San in treating gouty arthritis, which has a good therapeutic effect on gout inflammation, the proportion of Phellodendron bark, Magnolia bark, and Glycyrrhiza uralensis in the original Jin Huang San formula can be increased to further enhance the therapeutic effect of Jin Huang San-loaded microneedles on gouty arthritis.

[0018] Example 2 likeFigure 1 As shown, this embodiment is a further improvement on embodiment 1, as detailed below: Step 1: Researching the mechanism of action of Jinhuang Powder in treating gouty arthritis specifically includes: Step 1.1: Search for relevant target points of Jinhuang San in the treatment of gouty arthritis; databases such as Figure 2 As shown, these include GeneCards: a gene card database; DrugBank: a drug bank database; TTD: a therapeutic target database; PharmGKB: a pharmacogenomics knowledge base; and OMIM: an online human Mendelian genetics database. Step 1.2: Perform "medicinal material-target-disease" target analysis, such as... Figure 3 ; Step 1.3: Perform KEGG enrichment analysis, such as... Figure 4 To identify signaling pathways involved by differentially expressed genes; Step 1.4: By analyzing the distribution of differentially expressed genes in the KEGG pathway, the molecular mechanism by which Jin Huang San participates in the treatment of gouty arthritis was revealed. Step 1.5: Combining KEGG pathway enrichment analysis, it was determined that Jin Huang San can treat gouty arthritis through the TLR4 and NF-κB pathways. Step 1.6: Molecular docking of each component of Jinhuangsan with the TLR4 and NF-κB pathways was performed. It was finally determined that the three components, quercetin, β-sitosterol and stigmasterol, can directly act on the TLR4 and NF-κB pathways, with a docking score > 4.5. Step 1.7: Determine the distribution of quercetin, β-sitosterol, and stigmasterol in the three medicinal materials: Phellodendron bark, Magnolia bark, and licorice.

[0019] like Figure 5 A, quercetin docking results with TLR4 (docking score 5.0434); For example... Figure 5 D, quercetin docking results with NF-κB (docking score 7.2649); such as Figure 5 B, β-sitosterol docking results with TLR4 (dockingscore 6.2817); such as Figure 5 E, β-sitosterol docking results with NF-κB (docking score 5.4094); such as Figure 5 C, docking results of stigmasterol with TLR4 (docking score 5.6928); such as Figure 5F, docking results of stigmasterol with NF-κB (dockingscore 4.8381). Quercetin, β-sitosterol, and stigmasterol can dock with the TLR4 and NF-κB pathways, respectively, to treat gouty arthritis. Furthermore, it was determined that quercetin, β-sitosterol, and stigmasterol are distributed in three medicinal herbs: Phellodendron bark, Magnolia bark, and Licorice root, providing support for subsequent modifications to the original Jin Huang San formula.

[0020] Example 3 This embodiment is a further improvement on embodiment 2, specifically as follows: Step 2: Based on the research results, the proportions of the three medicinal materials, Phellodendron bark, Magnolia bark, and licorice, in the original Jin Huang San formula are increased to obtain the new Jin Huang San formula, which specifically includes: The original formula for Jinhuang San includes: 160g of turmeric, 160g of rhubarb, 160g of phellodendron bark, 64g of atractylodes rhizome, 64g of magnolia bark, 64g of tangerine peel, 64g of licorice root, 64g of raw arisaema rhizome, 160g of angelica root, and 320g of trichosanthes root. The adjusted formula for Jinhuang San is as follows: 80g turmeric, 80g rhubarb, 320g phellodendron bark, 32g atractylodes rhizome, 128g magnolia bark, 32g tangerine peel, 128g licorice root, 32g raw arisaema rhizome, 80g angelica root, and 160g trichosanthes root.

[0021] Example 4 This embodiment is a further improvement on embodiment 3, as detailed below: Step 3: Preparing Jin Huang San drug-loaded microneedles based on the new Jin Huang San formula specifically includes: Step 3.1: Decoction of the medicinal materials in the new prescription of Jinhuangsan to obtain Jinhuangsan freeze-dried powder; Step 3.2: Prepare Jin Huang San lyophilized powder into Jin Huang San drug-loaded microneedles.

[0022] Step 3.1, obtaining Jin Huang San freeze-dried powder from the medicinal materials in the new Jin Huang San formula, specifically includes: decocting the medicinal materials in the new Jin Huang San formula with water at a ratio of 1:9; centrifuging the resulting liquid and then filtering it to obtain a preliminary extract; allowing it to stand at room temperature for a period of time; then loading the preliminary extract into centrifuge tubes, centrifuging and filtering it again, and finally shaking it to obtain the final extract; pre-freezing the final extract for a period of time and then drying it to obtain Jin Huang San freeze-dried powder. The purified Jin Huang San freeze-dried powder has fewer impurities, facilitating the subsequent preparation of Jin Huang San drug-loaded microneedles.

[0023] The water used to decoct the herbs in the new formula for Jinhuang San is ultrapure water. This prevents contamination of the herbs and improves the quality of the medicine.

[0024] In specific implementation, the proportions of medicinal materials in the new formula of Jinhuang San were adjusted proportionally. The following herbs were used: 8g turmeric, 8g rhubarb, 32g phellodendron bark, 3.2g atractylodes rhizome, 12.8g magnolia bark, 3.2g dried tangerine peel, 12.8g licorice root, 3.2g raw arisaema rhizome, 8g angelica root, and 16g trichosanthes root, totaling 107.2g. Water was added at a 1:9 ratio and decocted to obtain approximately 1000ml of liquid. The liquid was centrifuged (12000 rpm, 15 minutes) and finely filtered to obtain a preliminary extract. After standing at room temperature for 4 hours, the filtered solution was aliquoted into centrifuge tubes, centrifuged again (10000 rpm, 5 minutes), finely filtered, and finally shaken for 10 minutes. The final extract was then obtained. Pre-freezing: Pre-freeze at -80℃ for 12-24 hours; then sublimation drying: Vacuum drying at -30℃ for 48-72 hours; finally, desorption drying: Dry at 30℃ for 24-48 hours. Freeze-drying is then complete, yielding golden loose freeze-dried powder.

[0025] Step 3.2: The preparation of Jin Huang San lyophilized powder into Jin Huang San drug-loaded microneedles specifically includes: dissolving Jin Huang San lyophilized powder in water at a ratio of 1:3 to prepare a drug solution; weighing a certain amount of sodium hyaluronate and adding it to the drug solution, while continuously stirring to obtain a mixed solution; centrifuging the mixed solution in a centrifuge to obtain a gel solution; injecting the gel solution into a microneedle mold; placing the microneedle mold in a filling device and vacuuming it; then placing the mold in a sleeve for centrifugation; drying and demolding.

[0026] Sodium hyaluronate should be added gradually to the drug solution. It dissolves easily and evenly.

[0027] In the specific implementation, weigh 0.6 g of sodium hyaluronate with a molecular weight of 60,000-100,000 and add it sequentially to 2 mL of drug solution (1:3 solution of lyophilized Jin Huang San powder in water). Each addition is accompanied by vigorous stirring with a 1 mL pipette tip to ensure complete dissolution. Centrifuge the mixture (3000 rpm, 3 minutes) to remove air bubbles. Add the resulting gel solution to the groove of the microneedle mold using a syringe, place it in a filling device, apply vacuum (0.09-0.1 MPa), and maintain this vacuum for 2 minutes. Place the mold in a 50 mL tube and centrifuge (4000 rpm, 3 minutes), scraping off any air bubbles from the surface. Then, statically dry at 35°C for 24 hours, and unmold to obtain the drug-loaded microneedles.

[0028] It also includes step 4, which involves morphological and physical property testing of the gold-infused drug-loaded microneedles.

[0029] Scanning with an optical electron microscope at 50, 70, 100, and 200x magnification, respectively, as shown below. Figure 7As shown in A, 7B, 7C, and 7D, the prepared golden powder drug-loaded microneedles have stable morphology. Pressure tests were performed on the golden powder drug-loaded microneedles, as shown in the table below. Compared with unloaded microneedles, the physical properties of the golden powder drug-loaded microneedles did not change adversely, meeting the transdermal drug delivery standard and ensuring the therapeutic effect of the golden powder drug-loaded microneedles.

[0030] Example 5 A golden powder-loaded microneedle for improving gouty arthritis is prepared using the method described above.

[0031] In practice, it is made by using molds as follows: Figure 6 As shown in Figure A, the overall parameters are: Array: 24 pins * 3 turns = 72 pins; Structure distribution area: diameter 4.5mm, center hole 3mm; Overall patch size: diameter 6mm; Figure 6 As shown in B, the single needle parameters are: column length 600μm; bottom side length 300μm; center-to-center spacing (drug loading depth) 500μm for the golden powder drug-loaded microneedles.

[0032] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of the present invention should be included within the protection scope of the present invention.

Claims

1. A method for preparing a golden powder-loaded microneedle for improving gouty arthritis, characterized in that, Includes the following steps: Step 1: Study the mechanism of action of Jinhuang San in treating gouty arthritis; Step 2: Based on the research results, increase the proportion of three medicinal materials, namely Phellodendron bark, Magnolia bark, and Licorice, in the original Jinhuang San formula, and decrease the proportion of other medicinal materials to obtain the new Jinhuang San formula; Step 3: Prepare Jinhuangsan drug-loaded microneedles based on the new Jinhuangsan formula.

2. The method for preparing the golden powder-loaded drug microneedles for improving gouty arthritis according to claim 1, characterized in that, Step 1, studying the mechanism of action of Jinhuang San in treating gouty arthritis, specifically includes: Step 1.1: Search for relevant targets of Jinhuang San in the treatment of gouty arthritis; Step 1.2: Perform "medicinal material-target-disease" target analysis; Step 1.3: Identify the signaling pathways involved by differentially expressed genes through KEGG enrichment analysis; Step 1.4: By analyzing the distribution of differentially expressed genes in the KEGG pathway, the molecular mechanism by which Jin Huang San participates in the treatment of gouty arthritis was revealed. Step 1.5: Combining KEGG pathway enrichment analysis, it was determined that Jin Huang San treats gouty arthritis through two pathways: TLR4 and NF-κB. Step 1.6: Molecular docking of each component of Jinhuangsan with the TLR4 and NF-κB pathways was performed, and it was finally determined that the three components, quercetin, β-sitosterol and stigmasterol, can directly act on the TLR4 and NF-κB pathways. Step 1.7: Determine the distribution of quercetin, β-sitosterol, and stigmasterol in the three medicinal materials: Phellodendron bark, Magnolia bark, and licorice.

3. The method for preparing the golden powder-loaded microneedles for improving gouty arthritis according to claim 1, characterized in that, Step 2, based on the research results, involves increasing the proportions of Phellodendron bark, Magnolia bark, and licorice in the original Jin Huang San formula to obtain a new Jin Huang San formula, which specifically includes: The original formula for Jinhuang San includes: 160g of turmeric, 160g of rhubarb, 160g of phellodendron bark, 64g of atractylodes rhizome, 64g of magnolia bark, 64g of tangerine peel, 64g of licorice root, 64g of raw arisaema rhizome, 160g of angelica root, and 320g of trichosanthes root. The adjusted formula for Jinhuang San is as follows: 80g turmeric, 80g rhubarb, 320g phellodendron bark, 32g atractylodes rhizome, 128g magnolia bark, 32g tangerine peel, 128g licorice root, 32g raw arisaema rhizome, 80g angelica root, and 160g trichosanthes root.

4. The method for preparing the golden powder-loaded microneedles for improving gouty arthritis according to claim 1, characterized in that, Step 3, preparing Jin Huang San drug-loaded microneedles based on the new Jin Huang San formula, specifically includes: Step 3.1: Decoction of the medicinal materials in the new prescription of Jinhuangsan to obtain Jinhuangsan freeze-dried powder; Step 3.2: Prepare Jin Huang San lyophilized powder into Jin Huang San drug-loaded microneedles.

5. The method for preparing the golden powder-loaded microneedles for improving gouty arthritis according to claim 4, characterized in that, Step 3.1, obtaining Jinhuangsan freeze-dried powder by decocting the medicinal materials in the new Jinhuangsan formula, specifically includes: decocting the medicinal materials in the new Jinhuangsan formula with water at a ratio of 1:9, centrifuging the decoction and then filtering it to obtain a preliminary extract, placing it at room temperature for a period of time, loading the preliminary extract into a centrifuge tube, centrifuging and filtering it, and finally shaking it to obtain the final extract, pre-freezing the final extract for a period of time and then drying it to obtain Jinhuangsan freeze-dried powder.

6. The method for preparing the golden powder-loaded microneedles for improving gouty arthritis according to claim 5, characterized in that, The water used to decoct the medicinal materials in the new prescription of Jinhuang San is ultrapure water.

7. The method for preparing the golden powder-loaded microneedles for improving gouty arthritis according to claim 5, characterized in that, Step 3.2, preparing Jin Huang San lyophilized powder into Jin Huang San drug-loaded microneedles, specifically includes: dissolving Jin Huang San lyophilized powder in water at a ratio of 1:3 to prepare a drug solution; weighing a certain amount of sodium hyaluronate and adding it to the drug solution, while continuously stirring to obtain a mixed solution; centrifuging the mixed solution in a centrifuge to obtain a gel solution; injecting the gel solution into a microneedle mold; placing the microneedle mold in a filling device and vacuuming it; then placing the mold in a sleeve for centrifugation; drying and demolding.

8. The method for preparing the golden powder-loaded microneedles for improving gouty arthritis according to claim 7, characterized in that, Sodium hyaluronate should be added to the drug solution one at a time.

9. The method for preparing the golden powder-loaded drug microneedles for improving gouty arthritis according to claim 1, characterized in that, It also includes step 4, which involves morphological and physical property testing of the gold-infused drug-loaded microneedles.

10. A golden powder-loaded microneedle for improving gouty arthritis, characterized in that, The golden powder-loaded microneedles are prepared using the method described in any one of claims 1 to 9 for preparing golden powder-loaded microneedles to improve gouty arthritis.

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