Composition and preparation for treating gouty arthritis as well as preparation method and application of composition and preparation

Abstract

The invention relates to the technical field of pharmaceutical compositions, and particularly provides a composition for treating gouty arthritis, which is prepared from the following components in parts by weight: 100 to 550 parts of gentiopicroside, 20 to 90 parts of tetrahydropalmatine, 0.5 to 2 parts of aconitine, 2000 to 2300 parts of 1, 2-propylene glycol and 2000 to 2300 parts of laurocapram. The invention also provides a preparation method of the pharmaceutical preparation for treating gouty arthritis, and the preparation method comprises the following steps: mixing the three effective components of gentiopicroside, tetrahydropalmatine and aconitine with 1, 2-propylene glycol and laurocapram, and matching with different auxiliary materials to prepare preparations of different dosage forms. The invention also provides an application of the composition in preparation of anti-inflammatory or analgesic drugs. The pharmaceutical preparation can be used for treating inflammatory pain caused by gouty arthritis, and has no toxic or side effect.

Description

Technical Field

[0001] The present invention relates to the technical field of pharmaceutical compositions, and in particular to a composition and preparation for treating gouty arthritis, and a preparation method and application thereof. Background Art

[0002] Gout is a metabolic disease caused by purine metabolism disorder and uric acid excretion disorder. Uric acid is the final product of purine metabolism and is usually excreted by the kidneys. Hyperuricemia occurs when factors such as genetic variation or pathological damage cause excessive purine metabolism to uric acid or reduced uric acid excretion. When excessive urate crystals are deposited in the joint cavity, joint capsule, kidneys and other parts, gout-related diseases such as gouty arthritis may develop. The typical symptoms of gouty arthritis include severe joint pain, swelling, redness, heat and tenderness of the skin, which usually occur suddenly, especially at night. The most common site of attack is the big toe joint, but it may also affect the knee, ankle and other joints. The main causes are related to genetics, eating habits, obesity, hypertension, poor renal function and certain drugs, which often cause patients unbearable pain, even motor dysfunction and decreased quality of life, which brings great pain to patients.

[0003] At present, the treatment of gouty arthritis inflammation and pain is divided into two methods: Chinese and Western medicine. Western medicine treatment includes uric acid-lowering drugs such as allopurinol and febuxostat, and anti-inflammatory drugs used in the acute phase such as non-steroidal anti-inflammatory drugs, colchicine, corticosteroids, etc. For example, diclofenac sodium can reduce the production of prostaglandins by inhibiting COX-2 enzymes or inhibiting both COX-1 and COX-2 enzymes, thereby alleviating pain and inflammation. As we all know, gouty arthritis is a chronic and difficult disease to cure. Although these drug treatments have good effects, they cannot be taken for a long time, and the toxic side effects are also obvious, often causing headaches, nausea, rashes and asthma, etc. In severe cases, it may even cause damage to gastrointestinal, liver and kidney functions.

[0004] In addition, Chinese medicine for the treatment of gouty arthritis also has great advantages, mainly because it has no obvious toxic and side effects; For example, the Chinese patent with publication number CN105998900A discloses "Chinese medicine for the treatment of gout and hyperuricemia and its preparation method", which is mainly composed of the following raw materials by weight: 15-135 parts of Coptis chinensis, 8.5-76 parts of Evodia rutaecarpa, 14-130 parts of Leonurus japonicus, 50-900 parts of Corydalis yanhusuo, 18-158 parts of Lophatherum gracile, and 6-114 parts of Coix lachryma-jobi. It can be seen that the patent discloses a Chinese medicine composition with the effects of dispelling wind and dampness, reducing swelling and relieving pain. In addition, the existing technology for the treatment of arthritis in Chinese medicine therapy is basically to find a suitable combination of different Chinese medicines, but there is no exploration of the effective ingredients of the Chinese medicines that play a therapeutic role and their combination to explore the therapeutic effect of gouty arthritis. Therefore, further finding its effective ingredients and their combination on the basis of the existing Chinese medicine combination is an important task for developing an effective drug combination for the treatment of gouty arthritis. Summary of the invention

[0005] Based on the above problems, the purpose of the present invention is to provide a composition, preparation and preparation method and application for treating gouty arthritis, which is a compound pharmaceutical composition mainly composed of gentiopicroside, tetrahydropalmatine, aconitine, 1,2-propylene glycol and laurocapram as effective chemical components, and plays a role in strengthening the body and eliminating evil, promoting blood circulation and removing blood stasis, softening and dispersing nodules, protecting bones and regenerating bones, relieving pain and calming the nerves, and deriving the optimal compatibility combination from the mass fractions of different components of the composition, with synergistic effects and improved efficacy, which can meet the clinical demand for combined medication for the treatment of inflammatory pain problems caused by gouty arthritis.

[0006] The embodiments of the present invention are implemented by the following technical solutions:

[0007] A composition for treating gouty arthritis comprises the following components by weight: 100-550 parts of gentiopicroside, 20-90 parts of tetrahydropalmatine, 0.5-2 parts of aconitine, 2000-2300 parts of 1,2-propylene glycol and 2000-2300 parts of laurocapram.

[0008] Furthermore, the following components are included by weight: 130 parts of gentiopicroside, 21 parts of tetrahydropalmatine, 0.6 parts of aconitine, 2200 parts of 1,2-propylene glycol, and 2200 parts of laurocapram.

[0009] Furthermore, the composition comprises, by weight: 261 parts of gentiopicroside, 43 parts of tetrahydropalmatine, 1.3 parts of aconitine, 2200 parts of 1,2-propylene glycol, and 2200 parts of laurocapram.

[0010] Furthermore, the composition comprises, by weight: 522 parts of gentiopicroside, 87 parts of tetrahydropalmatine, 1.9 parts of aconitine, 2200 parts of 1,2-propylene glycol, and 2200 parts of laurocapram.

[0011] Furthermore, the aconitine is derived from one or more of Chuanwu, processed Chuanwu, Fuzi, processed Fuzi, Kusnezoffii, processed Kusnezoffii, and Kusnezoffii leaves.

[0012] A pharmaceutical preparation for treating gouty arthritis comprises the above-mentioned composition and a pharmaceutically acceptable excipient.

[0013] Furthermore, the dosage form of the preparation can be any one of a gel, a spray, a film coating, an ointment or a patch.

[0014] Furthermore, the extraction of gentiopicroside is as follows: after the powder of Gentiana macrophylla is crushed to 80-100 mesh, it is extracted with 8-15 times the amount of a hydrophilic organic solvent for 3-5 hours, and the temperature is maintained at 30-60°C; after extraction, the filtrate is filtered and concentrated, and the solvent is recovered to obtain an extract; the extract is dried at 80-100°C until the moisture content does not exceed 20% to obtain a dry extract; the dry extract is mixed with 10-30 times the amount of a hydrophilic organic solvent and 20-40% by weight of activated carbon, and filtered after reflux decolorization; after the solvent is recovered under reduced pressure, crystals are precipitated and refrigerated for 3-5 days, and gentiopicroside crude crystals are obtained by filtration; the crude crystals are dissolved in a hydrophilic organic solvent, heated to saturation, and then hot-filtered, refrigerated for 1-2 days, and filtered again; repeating this crystallization process 3-5 times, gentiopicroside crystals with a purity of more than 98% can be obtained.

[0015] Furthermore, the extraction of tetrahydropalmatine: the chopped corydalis raw material is placed in a container, 1% hydrochloric acid is added, and soaked for 0 to 40 minutes to convert the alkaloids into salts for subsequent extraction. Then, an appropriate amount of sodium hydroxide is used to neutralize the alkaloids to free them; then, 95% ethanol is used to soak or reflux the extracted raw material, and the solid residue is filtered to remove the solid residue to obtain an extract containing tetrahydropalmatine. Then, a rotary evaporator is used to concentrate the extract; thin layer chromatography is used to separate and purify the concentrated extract, and finally, crystallization and drying are performed to obtain tetrahydropalmatine.

[0016] Further, the extraction of aconitine: after the aconitine source medicinal materials (one or more of Chuanwu, processed Chuanwu, Fuzi, processed Fuzi, Kusnezoffii, processed Kusnezoffii, and Kusnezoffii leaves) are crushed, they are mixed with three times the volume of industrial ethanol for 72 hours of cold extraction, and repeated three times to fully extract the effective ingredients. The extract is distilled under reduced pressure below 45°C to remove ethanol to obtain an extract. The extract is dissolved in acidified water adjusted to pH 3-4, and then the pH is adjusted to 9-10.5 with ammonia water, preferably to about 10, and liquid-liquid extraction is performed three times with chloroform to transfer alkaloids. After the extract is decompressed and desolventized, a crude extract of total alkaloids of Kusnezoffii is obtained; this crude extract is purified by macroporous adsorption resin, gradient elution is performed using distilled water and ethanol, and the ethanol elution component is concentrated under reduced pressure below 45°C, the precipitated light yellow solid is filtered out, and dried to finally obtain refined aconitine.

[0017] The present invention also provides a method for preparing a pharmaceutical preparation for treating gouty arthritis, comprising the following steps: mixing the three active ingredients obtained above with corresponding doses of 1,2-propylene glycol and laurocapram, and adding different excipients to prepare preparations of different dosage forms.

[0018] The present invention also provides an application of the above-mentioned composition in the preparation of drugs for anti-inflammatory, analgesic and uric acid-lowering treatment.

[0019] More specifically, the invention provides an application of the above-mentioned composition in the preparation of a medicine for treating gouty arthritis.

[0020] The technical solution of the embodiment of the present invention has at least the following advantages and beneficial effects:

[0021] The gentiopicroside, tetrahydropalmatine, aconitine, gentiopicroside and 1,2-propylene glycol of the present invention mainly reduce uric acid production and increase excretion in the treatment of gouty arthritis by regulating neurotransmitter receptor activity and G protein-coupled receptor signaling pathway, thereby reducing uric acid levels in the body. At the same time, they synergistically affect cell metabolism, regulate hormone response and cell signaling, reduce inflammatory response and pain; and by regulating blood circulation, muscle function and intracellular and extracellular material exchange, they act comprehensively on the treatment of gout. This comprehensive effect helps to alleviate gout symptoms, regulate uric acid metabolic balance, and improve the quality of life of patients without toxic side effects. BRIEF DESCRIPTION OF THE DRAWINGS

[0022] In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the drawings required for use in the embodiments are briefly introduced below. It should be understood that the following drawings only show certain embodiments of the present invention and therefore should not be regarded as limiting the scope. For ordinary technicians in this field, other related drawings can be obtained based on these drawings without creative work.

[0023] Figure 1 This is a schematic diagram of the results of using the kit to detect the uric acid content in the serum of mice in different experimental groups in the experimental examples of the present invention;

[0024] Figure 2 This is a schematic diagram showing the comparison of ankle swelling levels of mice in different experimental groups under different treatment conditions in the experimental examples of the present invention;

[0025] Figure 3 Schematic diagram of the comparison of the toe volumes of mice in different experimental groups under different treatment conditions in the experimental examples of the present invention; wherein, Figure 3 (A) is a schematic diagram of ankle joint swelling rate results. Figure 3 (B) for Figure 3 (A) Schematic diagram of the results of normalization of swelling rate;

[0026] Figure 4 This is a schematic diagram of the results of using the plantar thermal pain stimulation method to detect the pain threshold of mice in different treatment groups in the experimental examples of the present invention; wherein, Figure 4 (A) is a schematic diagram of the heat pain threshold results. Figure 4 (B) for Figure 4 (A) Schematic diagram of the results of normalization of heat pain threshold;

[0027] Figure 5 This is a schematic diagram of the results of using ELISA to detect the contents of four commonly used inflammatory indicators of gouty arthritis in mouse serum in the experimental examples of the present invention, wherein: Figure 5 (A)-(D) are schematic diagrams representing the results of the contents of IL-1β, IL-6, TNF-α, and IL-18, respectively;

[0028] Figure 6 This is a schematic diagram of the results of using ELISA to detect the contents of four commonly used inflammatory indicators of gouty arthritis in the ankle joint tissue of mice in the experimental examples of the present invention; Figure 6 (A)-(D) are schematic diagrams representing the results of the contents of IL-1β, IL-6, TNF-α, and IL-18, respectively;

[0029] Figure 7 Schematic diagram of the results of H&E staining of ankle joints of mice in different experimental groups in the experimental examples of the present invention to detect tissue damage and inflammatory cell infiltration;

[0030] Figure 8 (A) Figure 8 (B) are schematic diagrams of the results of using the kit to detect the contents of CRE and BUN in the serum of kidney-related indicators in mice in different experimental groups in the experimental examples of the present invention. DETAILED DESCRIPTION

[0031] In order to make the purpose, technical scheme and advantages of the embodiments of the present invention clearer, the technical scheme in the embodiments of the present invention will be described clearly and completely below. If the specific conditions are not specified in the embodiments, they are carried out according to conventional conditions or conditions recommended by the manufacturer. If the manufacturer of the reagents or instruments used is not specified, they are all conventional products that can be purchased commercially.

[0032] The following is a detailed description of a composition, a preparation, a preparation method and an application thereof for treating gouty arthritis provided in an embodiment of the present invention.

[0033] A composition for treating gouty arthritis comprises, by weight, 150-300 parts of gentiopicroside, 30-50 parts of tetrahydropalmatine, 0.5-1.5 parts of aconitine, 2.9-5.6 parts of 1,2-propylene glycol and 2.9-5.6 parts of laurocapram.

[0034] Gentiopicroside may effectively regulate carbohydrate metabolism and reduce the production of inflammatory mediators by inhibiting multiple enzymes, such as β-galactosidase (such as GLB1), O-glucosidase (such as OGA) and acid α-glucosidase (such as GAA). In addition, gentiopicroside can intervene in signal transduction, especially by regulating mitogen-activated protein kinase kinase 4 (MAP2K4), thereby controlling the inflammatory response. It may also act on hexosidases (such as HEXB) to further affect glucose and lipid metabolism, which is crucial for gout patients because increased blood sugar levels may aggravate gout symptoms. Tetrahydropalmatine may regulate gout-related biological processes by acting on multiple receptors, including activating or inhibiting opioid receptors (such as OPRD1), cholinergic receptors (such as CHRM1) and serotonin receptors (such as HTR2A), thereby regulating the release of neurotransmitters and inflammatory responses. Tetrahydropalmatine may also affect hormone receptors (such as ESR1) and ion channels (such as SCN5A), and the interaction of these molecules is crucial to the metabolic pathways, inflammatory processes, and pain signal transmission of gout. Aconitine may control intracellular signal transduction and inflammatory responses by affecting protein kinases and phosphatases (such as PPP3CA). At the same time, it adjusts metabolic pathways by inhibiting specific enzymes (such as SRD5A1), thereby reducing the production of proinflammatory metabolites. In addition, aconitine may also regulate receptors (such as AVPR1B) to regulate gout-related pain perception and inflammatory processes. 1,2-Propanediol may act on FUCA1 in the treatment of gout to regulate its activity to affect glucose metabolism and inflammatory response, thereby alleviating gout symptoms. Laurocapram indirectly regulates multiple signaling pathways and related enzyme activities by inhibiting microtubule polymerization, affecting the cytoskeleton and reducing the migration of inflammatory cells, such as inhibiting MAPK14 and ACACB, reducing the production of inflammatory mediators such as LTB4, and regulating the activity of NR3C1, thereby alleviating inflammation and pain caused by gout. In addition, both 1,2-propylene glycol and laurocapram play auxiliary roles in transdermal delivery. For example, 1,2-propylene glycol, as a transdermal delivery auxiliary, promotes drug absorption by increasing the fluidity of skin cell membranes, moisturizing and acting as a solvent, while laurocapram may enhance transdermal absorption efficiency by changing the skin barrier function and interacting with drug molecules. It can be seen that the active ingredient combination acts on multiple different targets to synergistically regulate the inflammatory and pain responses of gouty arthritis at different levels, showing potential synergistic enhancement capabilities.

[0035] The gentiopicroside, tetrahydropalmatine, aconitine, gentiopicroside and 1,2-propylene glycol of the present invention mainly reduce uric acid production and increase excretion in the treatment of gouty arthritis by regulating neurotransmitter receptor activity and G protein-coupled receptor signaling pathway, thereby reducing uric acid levels in the body. At the same time, they synergistically affect cell metabolism, regulate hormone response and cell signaling, reduce inflammatory response and pain; and by regulating blood circulation, muscle function and intracellular and extracellular material exchange, they act comprehensively on the treatment of gout, and this comprehensive effect helps to alleviate gout symptoms, regulate uric acid metabolic balance, and improve the quality of life of patients.

[0036] Example 1

[0037] This embodiment provides a composition for treating gouty arthritis, comprising, by weight: 130 parts of gentiopicroside, 21 parts of tetrahydropalmatine, 0.6 parts of aconitine, 2200 parts of 1,2-propylene glycol and 2200 parts of laurocapram;

[0038] This embodiment provides a gel preparation for treating gouty arthritis, and the preparation method thereof is as follows:

[0039] S1. First, wash the Radix Gentianae, Rhizoma Corydalis and Radix Aconiti Lateralis, remove the sand and other impurities, and dry them to constant weight for later use;

[0040] S2. Extraction of effective ingredients from traditional Chinese medicine:

[0041] (1) Extracting gentiopicroside: After the powder of Gentiana macrophylla is crushed to 80 mesh, it is extracted with 10 times the amount of a hydrophilic organic solvent for 4 hours, and the temperature is maintained at 50°C; after the extraction, the filtrate is filtered and concentrated, and the solvent is recovered to obtain an extract; the extract is dried at 90°C until the moisture content does not exceed 20%, thereby obtaining a dry extract; the dry extract is mixed with 20 times the amount of a hydrophilic organic solvent and 30% by weight of activated carbon, and filtered after reflux decolorization; after the solvent is recovered under reduced pressure, crystals are precipitated and refrigerated for 4 days, and gentiopicroside crude crystals are obtained by filtration; the crude crystals are dissolved in a hydrophilic organic solvent, heated to saturation, and then hot-filtered, refrigerated for 1 day, and filtered again; this crystallization process is repeated 4 times to obtain gentiopicroside crystals with a purity of 99%;

[0042] (2) Extracting corydalis ethyl: placing the chopped corydalis raw material in a container, adding 1% hydrochloric acid, soaking for 30 minutes, and then neutralizing with an appropriate amount of sodium hydroxide; then soaking the treated raw material in 95% ethanol, filtering to remove solid residues, and obtaining an extract containing corydalis ethyl; then concentrating the extract using a rotary evaporator; separating and purifying the concentrated extract using thin layer chromatography, and finally crystallizing and drying to obtain corydalis ethyl;

[0043] (3) Extracting aconitine: After grinding the aconite, mix it with three times the volume of ethanol and perform cold extraction for 72 hours, repeating three times; the extract is subjected to reduced pressure distillation below 45°C to remove ethanol to obtain an extract; the extract is dissolved in acidified water adjusted to pH 3, and then the pH is adjusted to 10 with ammonia water, and liquid-liquid extraction is performed three times with chloroform. After the extract is desolvated under reduced pressure, a crude extract of total alkaloids of aconite is obtained; the crude extract is purified by macroporous adsorption resin, using distilled water and 30% ethanol for gradient elution, and the ethanol elution component is concentrated under reduced pressure below 45°C, and the precipitated light yellow solid is filtered out and dried to finally obtain refined aconitine;

[0044] S3. The three active ingredients obtained by extraction are stirred with 790 parts of 60%-75% ethanol to dissolve, and then 1,2-propylene glycol and laurocapram are added, and 55,000 parts of distilled water and carbomer are added and stirred to dissolve, and the above solutions are mixed, mixed, allowed to stand, and filtered to obtain a gel.

[0045] Example 2

[0046] The difference between this embodiment and embodiment 1 is that: a composition for treating gouty arthritis, comprising, by weight: 261 parts of gentiopicroside, 43 parts of tetrahydropalmatine, 1.3 parts of aconitine, 2200 parts of 1,2-propylene glycol and 2200 parts of laurocapram.

[0047] Example 3

[0048] The difference between this embodiment and embodiment 1 is that: a composition for treating gouty arthritis, comprising, by weight: 522 parts of gentiopicroside, 87 parts of tetrahydropalmatine, 1.9 parts of aconitine, 2200 parts of 1,2-propylene glycol and 2200 parts of laurocapram.

[0049] Experimental example

[0050] 1.1 Experimental animals: Adult male specific pathogen-free (SPF) BALB / c mice (8 weeks, 23 ± 2 g) were purchased from Hunan Slake Jingda Experimental Animal Co., Ltd., Animal Production License No.: SCXK (Xiang) 2021-0002. All experiments were completed at the Experimental Animal Center of Northwest Agriculture and Forestry University. Mice were raised in specific sterile facility conditions for 7 days, with a humidity of 40-60%, a 12-h dark cycle, and free access to food and water under these conditions.

[0051] 1.2 Animal grouping

[0052] The experimental animals were divided into 6 groups, 10 in each group, namely:

[0053] Blank group; model group (i.e., gouty arthritis group); Voltaren group; sample group 1, sample group 2, and sample group 3; wherein, sample group 1 is the gel prepared in Example 1; sample group 2 is the gel prepared in Example 2; and sample group 3 is the gel prepared in Example 3.

[0054] 1.3 Gouty arthritis modeling: After adaptive feeding, BALB / c mice were anesthetized with isoflurane, the right hind ankle was shaved, and 20 μL of MSU crystals (0.5 mg) suspended in sterile PBS were injected into the tibiotarsal joint (ankle joint) of the mice. The modeling was successful when the ankle joint of the mice showed obvious swelling and thermal hyperalgesia 2 hours after the injection of MSU crystals. The blank group performed the same operation, but the ankle joint injection solution was sterile PBS; the other groups were all constructed with acute gouty arthritis models.

[0055] 1.4 Dosage method: 1 hour before modeling, the blank group applied an appropriate amount of drug carrier (transdermal penetration enhancer) to the right hind ankle; the Voltaren group applied the same volume of Voltaren emulsion; the sample groups 1, 2, and 3 applied the same volume of gel prepared in Example 1, Example 2, and Example 3, respectively. The corresponding drugs were applied again 4h, 9h, 13h, 23h, and 47h after modeling.

[0056] 1.5 Animal evaluation of gouty arthritis

[0057] In order to detect the inflammatory response of the ankle joint, a horizontal line was marked above the ankle joint of the mouse with a marker before modeling, which was used as the measurement basis for the subsequent swelling degree. Before modeling and 2h, 5h, 8h, 24h, and 48h after modeling, the volume below the ankle joint of the mouse was measured with a toe volume meter and the ankle swelling degree was calculated. Ankle swelling degree (%) = (ankle volume after modeling - ankle volume before modeling) / ankle volume before modeling × 100%.

[0058] 1.6 Plantar thermal pain stimulation method to detect pain threshold

[0059] The mice were placed in a foot heat pain meter, which was surrounded by transparent plastic and had a glass bottom. After the mice adapted to the environment for 5 minutes, a thermal radiation meter was used to emit a beam of light at the glass floor, and the time from the start of radiation to the withdrawal of the mouse's foot was recorded, which was the mouse's thermal withdrawal latency. The threshold of each group of mice was measured 1 hour before the injection of MSU as the pain threshold baseline, and the heat pain was measured multiple times at 2h, 5h, 8h, 24h, and 48h after the injection of MSU.

[0060] 1.7 Histopathological examination

[0061] Mice were killed 24 hours after MSU injection. The ankle joints were quickly cut off from the line above the ankle joints with sterile tissue scissors, fixed with 4% paraformaldehyde, stored in a 4°C refrigerator, and then decalcified with EDTA. After embedding with paraffin, the tissues were cut into 4-micron slices and stained with H&E. The stained slices were observed using an inverted microanalysis system to observe inflammatory cell infiltration and tissue damage.

[0062] 1.8ELISA test

[0063] Serum sample collection: Blood samples were collected 24 hours after MSU injection. After the mice were anesthetized with isoflurane, blood was collected from the eyeballs and placed in a tube without heparin coagulant. The mice were then killed by dislocating the neck. The blood was allowed to stand at room temperature for 1 hour, then centrifuged at 3000 r / min at room temperature for 10 minutes, and the supernatant was obtained as serum.

[0064] Tissue sample collection: Mice were killed 24 hours after MSU injection. The ankle joints were quickly cut off from the line above the ankle joints with sterile tissue scissors, and the tissues were rinsed with pre-cooled PBS to remove residual blood. After weighing, the tissues were cut into pieces with sterilized tissue scissors. Lysis solution was injected into the tissue samples according to the ratio of adding 150 to 250 microliters of RIPA lysis buffer for every 20 mg of tissue. Homogenization was performed using a glass tissue grinder until the tissue was completely broken. Finally, the tissue homogenate was aspirated into an EP tube, placed in a low-temperature high-speed centrifuge, and centrifuged at a speed of 10,000 to 14,000 g for 3 to 5 minutes. The supernatant was aspirated into a new EP tube.

[0065] The ELISA kit was operated according to the official instructions, and the standard curve was drawn to determine the levels of IL-1β, IL-6, TNF-α, and IL-18 in the supernatant.

[0066] 1.9 Kit detection

[0067] Serum sample collection: Blood samples were collected 24 hours after MSU injection. After the mice were anesthetized with isoflurane, blood was collected from the eyeballs and placed in a tube without heparin coagulant. The mice were then killed by dislocating the neck. The blood was allowed to stand at room temperature for 1 hour, then centrifuged at 3000 r / min at room temperature for 10 minutes, and the supernatant was obtained as serum.

[0068] According to the official instructions of the uric acid (UA), creatinine (CRE), and urea nitrogen (BUN) assay kit of Nanjing Jiancheng Bioengineering Institute, the standard curve was drawn and the levels of UA, CRE, and BUN in the supernatant were determined.

[0069] 1.10 Results Analysis

[0070] During a gout attack, serum uric acid levels usually rise dramatically. This is because during a gout attack, uric acid crystals are deposited in the joints, triggering an inflammatory response and also releasing more uric acid into the blood. Figure 1 The results showed that the blood uric acid level of mice in the gouty arthritis model group was significantly increased compared with that in the blank group (P<0.05); the Voltaren group and sample groups 1, 2 and 3 were able to reduce the blood uric acid index (P<0.05), and the blood uric acid indexes of sample groups 1, 2 and 3 were equivalent to those of the Voltaren group, indicating that the therapeutic effect of the gel prepared in the embodiment of the present invention is equivalent to that of Western medicine, and the effective ingredients of Chinese medicine can reduce the toxic and side effects caused by Western medicine.

[0071] The degree of ankle swelling can reflect the severity of gouty arthritis and the intensity of the inflammatory response. Figure 2 (Swelling of the right hind ankle of mice) and Figure 3 (A) (Ankle joint swelling rate) The results showed that 2 hours after MSU injection, the ankle joints of mice were significantly swollen and lasted until 48 hours, indicating that the model was successful. The swelling degree of the right ankle joints of mice in the gouty arthritis model group was significantly higher than that in the blank group, while the sample groups 1, 2 and 3 using Voltaren emulsion and different doses of the Chinese medicine active ingredient composition could alleviate the swelling degree of the ankle joints of mice at the corresponding time points. Figure 3 (B) for Figure 3 (A) The normalization of swelling rate aims to eliminate the influence of swelling rate at different time points on the overall swelling rate. The results show that compared with the control group, the paw swelling index of mice treated with Voltaren emulsion and different doses of the Chinese herbal effective ingredient composition was significantly reduced after MSU injection (P<0.05), indicating that the effects of each treatment group and the high-dose group of the Chinese herbal effective ingredient composition were equivalent to Voltaren, and had a significant alleviating effect on the swelling caused by gouty arthritis.

[0072] In gouty arthritis, uric acid crystals deposit in the joints, causing an inflammatory response that leads to damage to joint tissue and increased perception of pain. Figure 4 (A) The results of heat pain threshold showed that 2 hours after MSU injection, the time for mice to feel heat pain in their ankles was shortened, indicating that the model was successfully established. As time went on, the degree of heat pain in the right posterior ankle joints of mice in the gouty arthritis model group continued to decrease without significant improvement. However, the use of Voltaren emulsion and different doses of the Chinese medicine active ingredient composition in sample groups 1, 2 and 3 could improve the degree of heat pain in the right posterior ankle joints of mice within 48 hours. Figure 4(B) is the normalization treatment of the heat pain threshold of (A), the purpose of which is to eliminate the influence of the heat pain threshold at different time points on the overall heat pain threshold. The results showed that compared with the control group, the heat pain index of mice treated with Voltaren emulsion and different doses of the Chinese herbal effective ingredient composition was significantly increased after MSU injection (P<0.05), indicating that each treatment group had a significant improvement in the pain level of gouty arthritis, and the effect of the high-dose group of the Chinese herbal effective ingredient composition was equivalent to Voltaren.

[0073] Gouty arthritis is an inflammatory joint disease caused by the deposition of uric acid crystals in the joints. High uric acid levels are one of the main characteristics of gout, and uric acid crystals activate an inflammatory response, leading to symptoms such as joint pain, swelling, and redness. Figure 5 ((A) IL-1β, (B) IL-6, (C) TNF-α, (D) IL-1β in mouse serum) and Figure 6 ((A) IL-1β, (B) IL-6, (C) TNF-α, (D) IL-1β in the ankle joint tissue of mice) The results showed that the four inflammatory indicators in the serum and ankle joint tissue of the mice in the gouty arthritis model group were significantly increased compared with the blank group (P<0.05). The Voltaren emulsion group and sample groups 1, 2 and 3 were able to reduce these inflammatory indicators (P<0.05), indicating that each treatment group had a significant improvement effect on the degree of inflammation caused by uric acid crystals; and the effect indicators of the gel preparation prepared in the embodiment of the present invention in treating inflammation were comparable to those of the Voltaren emulsion group.

[0074] H&E staining can show inflammatory responses in tissues, including inflammatory cell infiltration and changes in joint structure. Figure 7 The results of H&E histopathological staining of the ankle joints of mice showed that there was no tissue structural damage and inflammatory cell infiltration in the ankle joints of mice in the normal group. The ankle joint tissues of mice in the gout model group were severely damaged, and there was obvious inflammatory cell infiltration. The Voltaren latex group showed reduced inflammatory cell infiltration. The number of inflammatory cells in sample groups 1, 2, and 3 was less than that in the model group, and the high-dose group had the least inflammatory cell infiltration; and in sample groups 1, 2, and 3, the high-dose group had fewer inflammatory cells than the Voltaren group.

[0075] During an acute gout attack, creatinine and urea nitrogen, two indicators of renal function, usually increase, partly because the inflammatory response during a gout attack is aggravated, which increases the burden on the kidneys and leads to temporary impairment of renal function. Figure 8The results of (A) creatinine and (B) urea nitrogen tests showed that the levels of blood creatinine and urea nitrogen in the gouty arthritis model group mice were significantly increased compared with the blank group (P<0.05). The Voltaren emulsion group and sample groups 1, 2, and 3 were able to reduce these indicators (P<0.05), indicating that Voltaren emulsion and the gel preparation prepared in the embodiment of the present invention can effectively alleviate the renal damage caused by acute gout.

[0076] The above are only preferred embodiments of the present invention and are not intended to limit the present invention. For those skilled in the art, the present invention may have various modifications and variations. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present invention shall be included in the protection scope of the present invention.

Claims

1. A composition for treating gouty arthritis, characterized in that: Calculated by weight, the invention comprises 100-550 parts of gentiopicroside, 20-90 parts of tetrahydropalmatine, 0.5-2 parts of aconitine, 2000-2300 parts of 1,2-propylene glycol and 2000-2300 parts of laurocapram.

2. The composition for treating gouty arthritis according to claim 1, characterized in that The composition comprises, by weight: 261 parts of gentiopicroside, 43 parts of tetrahydropalmatine, 1.3 parts of aconitine, 2200 parts of 1,2-propylene glycol and 2200 parts of laurocapram.

3. The composition for treating gouty arthritis according to claim 1, characterized in that The composition comprises, by weight: 522 parts of gentiopicroside, 87 parts of tetrahydropalmatine, 1.9 parts of aconitine, 2200 parts of 1,2-propylene glycol and 2200 parts of laurocapram.

4. The composition for treating gouty arthritis according to any one of claims 1 to 3, characterized in that: The extraction method of the gentiopicroside is: (1) crushing the powder of Gentiana macrophylla, extracting it with a hydrophilic organic solvent, and maintaining the extraction at a certain temperature for a period of time; (2) After extraction, filtering and concentrating the filtrate, recovering the solvent, and obtaining an extract; drying the extract until the moisture content does not exceed 20% to obtain a dry extract; (3) mixing the dry extract with a hydrophilic organic solvent and activated carbon, refluxing for decolorization, and then filtering; after recovering the solvent under reduced pressure, crystals are precipitated and refrigerated for a period of time, and then filtered to obtain crude gentiopicroside crystals; dissolving the crude gentiopicroside crystals in a hydrophilic organic solvent, heating to saturation, and then hot-filtering, refrigerating for a period of time, and filtering again, repeating this crystallization process multiple times to obtain gentiopicroside.

5. The composition for treating gouty arthritis according to any one of claims 1 to 3, characterized in that: The extraction method of corydalis butylene is as follows: chopped corydalis raw materials are placed in a container, hydrochloric acid is added, and the raw materials are soaked for a period of time, and then neutralized with an appropriate amount of sodium hydroxide; then the raw materials are soaked or refluxed for extraction with ethanol, and solid residues are filtered to obtain an extract containing corydalis butylene; and then the extract is concentrated, separated, purified, crystallized and dried to obtain corydalis butylene.

6. The composition for treating gouty arthritis according to any one of claims 1 to 3, characterized in that: The extraction method of aconitine comprises the following steps: crushing aconitine source medicinal material, mixing it with ethanol and cold soaking it for a period of time to obtain an extract; removing ethanol from the extract by vacuum distillation to obtain an extract; dissolving the extract in acidified water, adjusting the pH to 9-10.5, and extracting it multiple times to obtain an extract; decompressing the extract to obtain a crude extract of total alkaloids of Aconitum kusnezoffii, and then purifying it by adsorption resin, gradient eluting it, concentrating it under reduced pressure, filtering out the precipitated light yellow solid, and drying it to finally obtain refined aconitine.

7. A pharmaceutical preparation for treating gouty arthritis, characterized in that: The composition comprises the composition according to any one of claims 1 to 6 and a pharmaceutically acceptable excipient.

8. The pharmaceutical preparation for treating gouty arthritis according to claim 7, characterized in that: The dosage form of the preparation is any one of a gel, a spray, a film coating, an ointment or a patch.

9. A method for preparing a pharmaceutical preparation for treating gouty arthritis according to claim 7 or 8, characterized in that: The following steps are involved: Preparations of different dosage forms can be prepared by mixing gentiopicroside extract, corydaline extract, aconitine extract with 1,2-propylene glycol and laurocapram and adding different excipients.

10. Use of the composition according to any one of claims 1 to 6 in the preparation of drugs for anti-inflammatory, analgesic and uric acid-lowering treatment.

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