Pharmaceutical composition with analgesic effect as well as preparation method and application thereof

Abstract

The invention provides an external traditional Chinese medicine composition with a pain relieving effect. The external traditional Chinese medicine composition is an external preparation prepared from the following raw and auxiliary materials in parts by weight: 30-40 parts of prepared arisaema cum bile, 30-40 parts of baked ginger, 30-50 parts of radix angelicae, 20-30 parts of radix paeoniae alba, 15-18 parts of cinnamon, 6-8 parts of rhizoma bletillae, 15-20 parts of frankincense and 15-20 parts of myrrh. The invention further provides a preparation method and application of the external traditional Chinese medicine composition. Clinical application finds that according to dialectical medication of arthralgia, the external traditional Chinese medicine composition has a good effect on treating arthralgia and swelling of four limbs caused by wind, cold, dampness and blood stasis, joint cavity effusion or soft tissue diseases after trauma, and is convenient to operate and high in popularization degree.

Description

Technical Field

[0001] The invention relates to a pharmaceutical composition with analgesic effect and a preparation method and application thereof, belonging to the field of medicines. Background Art

[0002] Pain is a common symptom in clinical practice. It can be seen in many acute and chronic diseases. Patients suffer a lot of pain mentally, physically and mentally, which seriously affects their work and life. At present, Western medicine is mainly used to treat a variety of acute and chronic pain in clinical practice. One type is opioid analgesics, such as pethidine and morphine. Although these analgesics have a definite analgesic effect, long-term use will produce various side effects such as addiction and dependence, so they are strictly restricted in clinical application; the other type is non-opioid analgesics such as compound aspirin painkillers, but they are limited to the treatment of mild pain. Traditional Chinese medicine has a history of thousands of years in the treatment of pain syndrome. In the thousands of years of development history, Chinese medicine has been confirmed to have obvious and definite analgesic effects, and its toxic side effects are extremely small. Ma Fei San is a typical representative.

[0003] Arthralgia pain is a common disease in traditional Chinese medicine, and its etiology is mainly related to the invasion of exogenous evil and internal factors. Among them, wind, cold and dampness are common exogenous evils, and heat, fire, poison, etc. may also cause arthralgia pain. These exogenous evils will hinder the circulation of qi and blood, resulting in poor circulation of qi and blood, which in turn affects the normal function of the body. In addition, internal factors such as muscle and fascia strain, degenerative lesions of bones and joints can also cause arthralgia pain. Specifically, wind evil mainly causes obstruction of meridians and poor circulation of qi and blood, thus causing symptoms such as pain and cramps; cold and damp evil can cause swelling of tissues around joints, stagnation of moisture, as well as symptoms such as pain and numbness; damp and hot evil can cause joint redness, swelling, heat and pain. In addition, the etiology also includes internal factors, such as insufficient qi and blood, weak spleen and stomach, insufficient liver and kidney, etc. These problems can cause imbalance of yin and yang in the human body, thus causing arthralgia pain. The main methods and principles for treating pain are as follows: 1. Dispelling wind and cold: For arthralgia pain caused by external evils such as wind and cold, the method of dispelling wind and cold is used for treatment, such as warming and dispersing cold, activating collaterals and dispelling wind, etc., which can dispel cold evil, dredge meridians, relieve symptoms such as pain and cramps; 2. Dispelling dampness and resolving phlegm: For arthralgia pain symptoms caused by dampness, the method of dispelling dampness and resolving phlegm is used for treatment, such as warming and resolving dampness, resolving dampness and removing phlegm, etc. Chinese medicine can promote the discharge of moisture, reduce swelling and relieve pain; 3. Clearing away heat and detoxifying: For arthralgia pain caused by heat, fire, poison and other factors, the method of clearing away heat and detoxifying is used for treatment. Prescriptions such as clearing away heat and detoxifying, detoxifying and killing insects can clear heat and toxins from the body and relieve pain and symptoms; 4. Invigorating Qi and Strengthening the Spleen: For arthralgia pain caused by internal factors such as insufficient Qi and blood, weak spleen and stomach, the method of invigorating Qi and strengthening the spleen should be used for treatment, such as prescriptions such as Bu Zhong Yi Qi, Jian Pi He Wei, etc., which can regulate the balance of yin and yang in the body and enhance physical fitness; 5. Invigorating the Liver and Kidneys: For arthralgia pain caused by liver and kidney deficiency, the method of invigorating the liver and kidneys should be used for treatment, such as Chinese medicines such as nourishing the kidney and benefiting the liver, and nourishing the liver and kidneys.

[0004] Application No. 201210398599.X, Invention Name: A Chinese medicine for treating senile knee joint degeneration and its preparation method, the disclosed formula is "4.5 mass parts of processed Chuanwu, 4.5 mass parts of processed Caowu, 3 mass parts of Gallbladder Arisaema, 2 mass parts of Rhubarb, 3 mass parts of Red Peony Root, 3 mass parts of Angelica Dahurica, 1-1.5 mass parts of Cinnamon Bark, 1-2 mass parts of Ginger", the patent formula has the effect of curing and preventing recurrence of wind-cold-damp arthralgia such as early senile knee joint degeneration; it has the effect of curing or alleviating pain or even curing wind-cold-damp arthralgia such as severe stage senile knee joint degeneration. It is mainly aimed at specific populations and indications, and it is difficult to meet the needs of arthralgia pain caused by various clinical raw materials.

[0005] At present, Chinese herbal compound external plasters are highly safe, have a wide range of applications, are convenient and easy to use, and can often play a synergistic role with modern medicine; however, Chinese herbal compound external analgesics face the problems of unstable efficacy and narrow indications. Summary of the invention

[0006] The present invention provides a pharmaceutical composition with analgesic effect, and also provides a preparation method and application of the pharmaceutical composition.

[0007] The present invention provides an external Chinese medicine composition with analgesic effect, which is an external preparation prepared from raw materials and auxiliary materials in the following weight ratios:

[0008] 30-40 parts of processed Arisaema consanguineum, 30-40 parts of roasted ginger, 30-50 parts of Angelica dahurica, 20-30 parts of white peony root, 15-18 parts of cinnamon bark, 6-8 parts of Bletilla striata, 15-20 parts of frankincense and 15-20 parts of myrrh.

[0009] More preferably, it is an external preparation prepared from raw materials and auxiliary materials in the following weight ratios:

[0010] 30 parts of processed Arisaema, 40 parts of roasted ginger, 40 parts of Angelica dahurica, 30 parts of white peony root, 15 parts of cinnamon bark, 6 parts of Bletilla striata, 15 parts of frankincense, and 15 parts of myrrh.

[0011] The pharmaceutical composition of the present invention is prepared from the original powders and water or organic solvent extracts of processed Arisaema consanguineum, roasted ginger, angelica dahurica, white peony root, cinnamon bark, Bletilla striata, frankincense, myrrh and processed Aconitum kusnezoffii as active ingredients, and is added with pharmaceutically acceptable excipients or auxiliary ingredients to form a commonly used external preparation in medicine.

[0012] Among them, the external preparations are ointments, cataplasms, sprays, film-forming agents, creams, powders, oils, and tinctures.

[0013] The present invention also provides a method for preparing the external Chinese medicine composition, which comprises the following steps:

[0014] It includes the following steps:

[0015] 1) Weigh the raw materials of each weight ratio and grind into powder;

[0016] 2) adding water to mix, and adding pharmaceutically acceptable excipients or auxiliary ingredients to prepare a commonly used external preparation in medicine.

[0017] Further preferably, the external preparation is an ointment, and the preparation method of the ointment is:

[0018] 1) Weigh the raw materials of each weight ratio and grind them into fine powder using a low-temperature freezing mill;

[0019] 2) Take the powder and mix it with boiling water and vaseline in a specific ratio of 1:1.5:0.1. The ointment should not adhere to the wall;

[0020] 3) Apply the above-mentioned plaster on gauze and cover it with gauze; apply it on the affected area.

[0021] The present invention also provides use of the external Chinese medicine composition in preparing medicine for treating arthralgia pain.

[0022] The medicine is used to treat pain and swelling in the joints of the limbs, joint effusion or soft tissue diseases after trauma caused by wind, cold, dampness and blood stasis.

[0023] Wherein, the medicine is a medicine for treating knee osteoarthritis combined with synovitis.

[0024] The drug is a drug that reverses cartilage damage in knee osteoarthritis, inhibits TNF signaling pathway, reduces cell apoptosis, and slows down the progression of arthritis.

[0025] The medicine is used to treat diffuse swelling, pain, joint dysfunction or post-traumatic soft tissue diseases caused by wind, cold, dampness and blood stasis in the joints of the limbs.

[0026] Wind, cold, dampness, and blood stasis leading to pain and swelling in the limbs joints, effusion in the joint cavity, or soft tissue diseases after trauma belong to the category of "arthralgia" in traditional Chinese medicine. In the Qing Dynasty, Lin Peiqin's "Classification of Symptoms and Treatments·Theory of Arthralgia" said: "If arthralgia lasts for a long time, there will be phlegm, dampness, blood stasis, and stagnation in the meridians"; Zhao Xianke's "Medical Guide" said: "Phlegm, blood, and water are all the same thing." Phlegm and blood stasis are both the pathogenesis and the pathological characteristics of arthralgia; phlegm and blood stasis are intertwined, the meridians are blocked, the skin, tendons and bones are not nourished, and they penetrate into the joints to cause joint swelling, pain, deformity and movement disorders.

[0027] The external medicine composition of the present invention has the advantages of wide indications, and can be used for diseases caused by wind, cold, dampness, blood stasis, phlegm, etc., either alone or in combination. The composition comprises roasted ginger, processed gall bladder arisaema, white peony root, angelica root, cinnamon bark, frankincense, myrrh, and bletilla striata. In the formula, processed Arisaema, Angelica dahurica and Roasted Ginger are the three main herbs, which can dispel wind and cold, resolve phlegm and remove dampness; processed Arisaema is pungent, bitter and laxative, and can dry out dampness and resolve phlegm. "Compendium of Materia Medica" says: "... the nature of Arisaema is dry and quite urgent... Arisaema is inferior but good at moving... If there is wind phlegm, dampness and phlegm, and urgent saliva and phlegm, only Arisaema can disperse it." Angelica dahurica is pungent, dispersing, warming and dry, and aromatic, and it is good at dispersing wind and cold, removing dampness and relieving pain; "Compendium of Materia Medica" says: "... Angelica dahurica is extremely fragrant, can dispel wind and dampness, and its texture is extremely smooth, can harmonize and benefit blood vessels without drying up, so it is beneficial rather than harmful when used." Roasted Ginger is pungent and hot, dispersing cold and removing dampness, and "Depei Materia Medica" says: "Roasted Ginger is guarding but not "It can warm the liver meridian, remove evil and create new things, make yang grow and yin grow..."; Frankincense and myrrh are used together to promote blood circulation, relieve pain, and help the main medicine to dispel cold, eliminate dampness and relieve pain. Zhang Xichun said: "Frankincense and myrrh, the two medicines are used together... Their ability to promote ventilation and promote blood circulation is also good for treating wind-cold and dampness, numbness of the whole body, paralysis of the limbs and all kinds of sores, carbuncles and swellings... It can be used as powder for external application to sores, which can detoxify, reduce swelling, promote tissue regeneration and relieve pain. Although it is an open-circulation product, it does not consume qi and blood. It is indeed a good medicine." White peony root and white ginseng are adjuvants with moist texture and cold nature. They can nourish blood and harmonize meridians, soften tendons and relieve pain, but also prevent the main and auxiliary medicines from dispersing too much; cinnamon is pungent, sweet and hot, which can lead the medicine into the blood, dredge blood vessels and benefit joints.

[0028] In summary, the drug prescription of the present invention is well matched and has a wide range of indications. Among them, processed Arisaema ciliata and Rhizoma Zingiberis are used as the main drugs to dispel cold, dry dampness, resolve phlegm, restore yang and dredge the meridians; Angelica dahurica is pungent and disperses, helping Arisaema ciliata to dry dampness and resolve phlegm; frankincense and myrrh are dispersed in a direction, helping Rhizoma Zingiberis to dredge the meridians and activate blood circulation, promote qi and relieve pain, and are all ministerial drugs; white peony root is used as an adjuvant drug to prevent the above-mentioned drugs from dispersing too much; cinnamon bark and processed Aconitum kusnezoffii are meridian-inducing drugs to help open the pores and dispel cold and dampness; Bletilla striata harmonizes various drugs to moisturize the skin and promote muscle growth. In clinical applications, it is found that according to the dialectical medication of arthritis, the application of the external Chinese medicine composition of the present invention to treat wind, cold, dampness, blood stasis to pain and swelling of the limbs joints, joint cavity effusion or soft tissue diseases after trauma has a good effect, is easy to operate, and has a high degree of promotion. BRIEF DESCRIPTION OF THE DRAWINGS

[0029] Figure 1 Venn diagram of the intersection of Huiyang Pain Relief Cream drug targets and knee osteoarthritis-related targets;

[0030] Figure 2 'Combination - drug - active ingredient - potential target - disease' network diagram;

[0031] Figure 3 The protein interaction network diagram of 129 potential therapeutic genes;

[0032] Figure 4The protein interaction network diagram of 39 key targets (the circle in the center represents the top ten core genes screened by CytoHubba and ranked by degree value);

[0033] Figure 5 Gene Ontology (GO) pathway enrichment analysis (BP = biological process, CC = cellular component, MF = molecular function);

[0034] Figure 6 Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis;

[0035] Figure 7 Molecular docking diagram ((A) TNF-β-β-sitosterol; (B) TNF-β-stearyl sterol; (C) AKT1-stearyl sterol; (D) IL-1β-β-sitosterol);

[0036] Figure 8 HE staining of New Zealand rabbit articular cartilage (A: blank group, B: model group, C: Huiyang Pain-Relieving Ointment group, D: diclofenac diethylamine emulsion group);

[0037] Fig. 9 Expression of related proteins. DETAILED DESCRIPTION

[0038] Example 1 Preparation of the medicinal ointment of the present invention

[0039] 1) Weigh the raw materials of each weight ratio: 30g of processed Arisaema consanguineum, 40g of roasted ginger, 40g of Angelica dahurica, 30g of white peony root, 15g of cinnamon bark, 6g of Bletilla striata, 15g of frankincense, and 15g of myrrh, and grind them into fine powder using a low-temperature freezing mill;

[0040] 2) Take the powder and mix it with boiling water and vaseline in a specific ratio of 1:1.5:0.1. The ointment should not adhere to the wall;

[0041] 3) Apply the above-mentioned plaster on gauze and cover it with gauze; apply it on the affected area.

[0042] Example 2 Use of the medicinal spray of the present invention

[0043] Take the following raw materials in different weight ratios: 30g of processed Arisaema consanguineum, 30g of roasted ginger, 30g of Angelica dahurica, 20g of white peony root, 15g of cinnamon bark, 15g of Bletilla striata, 15g of frankincense and 15g of myrrh, add water to decoct for extraction and concentration, and add auxiliary materials of commonly used sprays to prepare a spray.

[0044] Example 3 Use of the medicinal powder of the present invention

[0045] Take the raw materials in different weight proportions: 40g processed Arisaema consanguineum, 40g roasted ginger, 50g Angelica dahurica, 30g white peony root, 18g cinnamon bark, 8g Bletilla striata, 20g frankincense, and 20g myrrh, grind them into powder directly, add boiling water to make a paste before use, and apply to the affected area.

[0046] The beneficial effects of the present invention are demonstrated by specific clinical trials as follows.

[0047] Test Example 1 Clinical trial of the drug of the present invention

[0048] 1. Methods:

[0049] 1.1 Diagnostic criteria

[0050] Refer to the Expert Consensus on Diagnosis and Clinical Efficacy Evaluation of Adult Knee Synovitis organized by the Orthopedics Branch of the Chinese Association of Traditional Chinese Medicine (Chinese Medical Association Orthopedics Branch, 2016)

[0051] Young and middle-aged people with a history of trauma or strain;

[0052] Middle-aged and elderly people, especially those who are obese;

[0053] Swelling of the knee joint;

[0054] Discomfort or pain in the knee joint;

[0055] When the knee joint is straightened or fully flexed, there is a feeling of fullness, discomfort or pain that is significantly aggravated;

[0056] Skin temperature may increase upon palpation and fluctuate upon pressing;

[0057] The tenderness point is not fixed, and the tenderness may be at the primary injury site;

[0058] Positive floating patella test, or B-ultrasound or MRI examination showing excessive knee joint effusion;

[0059] Quadriceps atrophy;

[0060] The joint puncture fluid is yellow or light yellow liquid with no fat droplets on the surface.

[0061] Experts recommend: If a person meets two of 3) and 4) and any two of the other two, he or she can be diagnosed with knee synovitis.

[0062] 1.2 Inclusion criteria

[0063] Meet the diagnostic criteria for type I knee synovitis;

[0064] 40 ≤ ≤ 55 years old, regardless of gender;

[0065] Visual Analogue Scale (VAS) score for knee pain: 2 points ≤ VAS ≤ 6 points;

[0066] Knee osteoarthritis Kellgren & Lawrence grade ≤ grade II;

[0067] The patients voluntarily participated in this study and read and signed the informed consent form.

[0068] 1.3 Exclusion criteria

[0069] Patients with other osteoarthritis, such as hip osteoarthritis, hand osteoarthritis, etc.;

[0070] Patients with secondary knee osteoarthritis caused by other diseases, such as secondary to joint trauma, congenital

[0071] or genetic diseases, etc.;

[0072] Those who had undergone knee joint cavity treatment within 1 month;

[0073] Skin ulceration, rash and infection around the knee joint;

[0074] Those with allergies or allergies to study drugs;

[0075] Women who are pregnant, trying to become pregnant or breastfeeding;

[0076] A randomized, double-blind, placebo-controlled, superiority trial design was adopted, and 60 cases were included and divided into a test group (hereinafter referred to as: Huiyang Pain Relief Ointment, prepared in Example 1) and a placebo group (Huiyang Pain Relief Ointment Simulator) at a ratio of 1:1. After 2 weeks of treatment, the changes in WOMAC scores and TCM syndrome scores before and after treatment in the three groups were observed, and the clinical efficacy was determined by referring to the "Standards for Diagnosis and Efficacy of TCM Diseases and Syndromes".

[0077] 2. Test results:

[0078] This trial included 60 patients who met the inclusion criteria, and 2 patients dropped out; they were randomly divided into the experimental group and the control group, and the WOMAC scores and TCM syndrome scores were recorded before treatment and on the 7th and 14th days after treatment. The results showed that the WOMAC scores and TCM syndrome scores before treatment and on the 7th and 14th days after treatment between the two groups were compared, P<0.05, and the difference was statistically significant. The total effective rate of the experimental group was 68.97%, and the total effective rate of the control group was 31.03%, P=0.004, and the difference was statistically significant.

[0079] Table 1 Analysis of the effective rate of the two groups of patients after 14 days of treatment

[0080]

[0081] Table 2 WOMAC scores and TCM symptom scores of the two groups of patients before treatment and 14 days after treatment (Mean±SD, n=58)

[0082]

[0083] Experimental Example 2 Molecular Mechanism of Action of the Traditional Chinese Medicine External Preparation Huiyang Pain-Relieving Ointment of the Present Invention on KOA Rabbits

[0084] 1. Materials and methods

[0085] Collection of drug active ingredients and targets:

[0086] The active ingredients of Huiyang Pain-Relieving Ointment were screened using the TCMSP database, and the target information was corrected using the UniProt database.

[0087] (1) Disease target collection:

[0088] KOA-related target genes were collected through GeneCards, OMIM and TTD databases.

[0089] (2) Construction of compound prescription-active drug ingredients-potential therapeutic genes-disease network:

[0090] The network diagram of Huiyang Zhitong Ointment and KOA was constructed by using Venny online tool and Cytoscape software.

[0091] (3) Construction of protein interaction network diagram:

[0092] The String database was used to construct a PPI network diagram and screen core genes.

[0093] (4) Enrichment analysis:

[0094] Oebiotech cloud platform was used to perform GO and KEGG pathway analysis on key genes.

[0095] (5) Molecular docking:

[0096] The binding ability between the main components of Huiyang Pain Relief Ointment and related target genes was analyzed by molecular docking.

[0097] (6) Animal experiments:

[0098] The efficacy of Huiyang Pain-Relieving Ointment was evaluated using the New Zealand rabbit model, and the pathological changes of knee joint tissue were detected using H&E staining and Western blot.

[0099] 2. Research Results

[0100] (1) Data on active compounds and targets of Huiyang Pain Relief Cream:

[0101] The TCMSP database was used to screen the active ingredients in Huiyang Pain Relief Paste, involving multiple Chinese herbal medicines (such as roasted ginger, processed Arisaema consanguineum, white peony root, etc.), and 1,627 drug targets were collected, and 238 related genes were obtained after deduplication.

[0102] (2) Knee osteoarthritis (KOA) disease targets:

[0103] A total of 2884 KOA-related targets were screened through GeneCards, OMIM and TTD databases.

[0104] (3)Integration of drug-disease target data:

[0105] Use Venny tool to integrate the active ingredients of Huiyang Pain Relief Cream with KOA disease target data ( Figure 1 ), and obtained an intersection of 138 active ingredients.

[0106] (4) Huiyang Pain Relief Ointment - Drug - Active Ingredient - Potential Therapeutic Target - Disease Network Construction:

[0107] A network including Huiyang Pain Relief Ointment drugs, active ingredients, potential therapeutic targets and diseases was constructed using Cytoscape software ( Figure 2 ). Figure 2 Huiyang Pain Relief Paste (HYZTP) 'compound-drug-active ingredient-potential target-disease' network diagram for knee osteoarthritis (KOA). Yellow octagons represent traditional Chinese medicines, blue circles represent active ingredients in traditional Chinese medicines, red hexagons represent common active ingredients in multiple traditional Chinese medicines, and green diamonds represent potential targets of Huiyang Pain Relief Paste for the treatment of knee osteoarthritis. KOA = knee osteoarthritis, HYZTP = Huiyang Pain Relief Paste.

[0108] (5) Protein interaction network diagram and gene screening:

[0109] 129 potential therapeutic targets were analyzed by String tool ( Figure 3 ), and screened out 39 key genes, and further screened out core genes such as IL6, AKT1, etc. through Cytoscape ( Figure 4 ).

[0110] (6) GO and KEGG enrichment analysis:

[0111] The GO analysis results of 39 key targets showed that there were 3101 entries in total, including 2308 biological processes (BP), 492 molecular functions (MF) and 300 cellular components (CC). The top 10 entries of BP, CC and MF were selected by P value calculation ranking ( Figure 5 ).

[0112] The KEGG analysis results of 39 key targets showed that there were 286 related pathways in total, and the top 20 most relevant pathways were obtained in descending order of enrichment scores ( Figure 6 ).

[0113] (7) Molecular docking:

[0114] The important chemical components of Huiyang Pain Relief Paste (beta-sitosterol, Stigmasterol) and key core genes related to enrichment analysis were selected for molecular docking (IL-1B, AKT1, TNF). When the binding energy is less than 0kJ / mol, the small molecule ligand can spontaneously bind to the protein receptor. If the binding energy is less than -5.0kJ / mol, it indicates that the binding ability of the two is better. The docking simulation generated 6 docking results, which are close to or less than -5kJ / mol binding energy, which means better binding ability.

[0115] Table 3 Binding energy of 1-molecule docking (kJ / MOL)

[0116]

[0117] According to the molecular docking results, it can be predicted that compounds such as (beta-sitosterol, Stigmasterol) may play an important role in the treatment of knee osteoarthritis with Huiyang Pain Relief Paste. Finally, the Pymol software was used to draw the best conformation picture ( Figure 7 ).

[0118] (8) HE staining:

[0119] Four weeks after administration, the knee joint tissues of the animals were collected for HE staining. The results showed that: blank group: no obvious pathological changes; model group: the articular cartilage surface was not smooth, the damage was obvious, the cartilage layer was thinned, the tide line was destroyed, the chondrocytes were arranged in disorder, and many vacuoles and inflammatory infiltrations were visible. Diclofenac diethylamine emulsion group and Huiyang analgesic ointment group: the articular cartilage surface was occasionally damaged, not smooth, the tide line existed, the chondrocytes were arranged more neatly, and the number of proliferating chondrocytes was visible. The results show that Huiyang analgesic ointment can effectively reverse the cartilage damage in knee osteoarthritis ( Figure 8 ).

[0120] (9) Western Blot:

[0121] In order to verify the results of network pharmacology, the present invention tested the expression of related proteins in the TNF signaling pathway and apoptosis signaling pathway. Western blot images and corresponding density analysis showed that compared with the blank group, the expression levels of TNF-α, RIP1, CASP3, and BAX proteins in the model group increased significantly; but after treatment with Huiyang Pain Relief Ointment, the expression levels of these proteins decreased significantly ( Fig. 9BE). The experimental results showed that Huiyang Pain-Relieving Ointment may inhibit TNF signaling pathway, reduce cell apoptosis and slow down the progression of OA.

[0122] The protein expressions of TNF-α, RIP1, CASP3 and BAX were detected by Western blot. Fig. 9 A). Normalized TNF-α ( Fig. 9 B) RIP1 Fig. 9 C), CASP3( Fig. 9 D) and BAX( Fig. 9 E) was quantitatively analyzed. In the quantitative graph, the green circle represents the blank group, the red circle represents the model group, and the orange circle represents the HYZTP group. The data are expressed as mean ± standard deviation. *p<0.05, **p<0.01

[0123] 3. Conclusion

[0124] The present invention explores the possible mechanism of action of Huiyang Pain Relief Plaster (HYZTP) in the treatment of knee osteoarthritis (KOA), and combines network pharmacology and molecular docking technology to reveal its potential therapeutic pathway. By integrating drug and disease target information, it is found that 138 active ingredients in HYZTP may be related to the treatment of KOA, especially beta-sitosterol, Stigmasterol and daidzein, which show significant therapeutic effects in the treatment of osteoarthritis. These compounds exert therapeutic effects through mechanisms such as anti-oxidation, anti-inflammatory and inhibition of joint degeneration.

[0125] Molecular docking results showed that beta-sitosterol and Stigmasterol had good binding ability with key hub gene-related proteins such as TNF-α, IL-1β and AKT1, which may be the material basis for HYZTP to treat KOA. H&E staining and Western Blot experiments further verified that HYZTP can promote chondrocyte proliferation, reverse articular cartilage damage, and reduce cell apoptosis by inhibiting the TNF signaling pathway, thereby slowing down the progression of OA.

[0126] Through GO and KEGG enrichment analysis, it was confirmed that the biological processes involved in HYZTP treatment of KOA include inflammatory response, oxidative response, cell apoptosis, etc., and the key signaling pathways include TNF signaling pathway, IL-17 signaling pathway and Toll-like receptor signaling pathway. These pathways are closely related to the pathological mechanism of KOA, especially the TNF signaling pathway plays a core role in regulating inflammatory response and cell apoptosis.

[0127] In general, HYZTP may slow down the progression of KOA by regulating the TNF signaling pathway, reducing inflammatory factors and cell apoptosis. This finding provides a new perspective for exploring the application of Huiyang Pain-Relieving Ointment in the treatment of OA in the future and lays the foundation for further experimental research.

Claims

1. A Chinese medicine composition for external use having analgesic effect, characterized in that: It is an external preparation made from the following raw materials and auxiliary materials in the following weight ratios: 30-40 parts of processed Arisaema consanguineum, 30-40 parts of roasted ginger, 30-50 parts of Angelica dahurica, 20-30 parts of white peony root, 15-18 parts of cinnamon bark, 6-8 parts of Bletilla striata, 15-20 parts of frankincense and 15-20 parts of myrrh.

2. The external Chinese medicine composition according to claim 1, characterized in that: It is an external preparation made from the following raw materials and auxiliary materials in the following weight ratios: 30 parts of processed Arisaema, 40 parts of roasted ginger, 40 parts of Angelica dahurica, 30 parts of white peony root, 15 parts of cinnamon bark, 6 parts of Bletilla striata, 15 parts of frankincense, and 15 parts of myrrh.

3. The external Chinese medicine composition according to claim 1 or 2, characterized in that: The invention is prepared from the original powders of processed Arisaema consanguineum, roasted ginger, angelica dahurica, white peony root, cinnamon bark, Bletilla striata, frankincense and myrrh, water or organic solvent extracts as active ingredients, and pharmaceutically acceptable excipients or auxiliary ingredients are added to prepare a commonly used external preparation in medicine.

4. The external Chinese medicine composition according to claim 3, characterized in that: The external preparations are ointments, cataplasms, sprays, film formers, creams, powders, oils, and tinctures.

5. The method for preparing the external Chinese medicine composition according to any one of claims 1 to 5, characterized in that: It includes the following steps: 1) Weigh the raw materials of each weight ratio and grind into powder; 2) adding water to mix, and adding pharmaceutically acceptable excipients or auxiliary ingredients to prepare a commonly used external preparation in medicine.

6. The method for preparing the external Chinese medicine composition according to claim 5, characterized in that: The external preparation is an ointment, and the preparation method of the ointment is: 1) Weigh the raw materials of each weight ratio and grind them into fine powder using a low-temperature freezing mill; 2) Take the powder and mix it with boiling water and vaseline in a specific ratio of 1:1.5:0.

1. The ointment should not adhere to the wall; 3) Apply the above-mentioned plaster on gauze and cover it with gauze; apply it on the affected area.

7. Use of the external Chinese medicine composition according to any one of claims 1 to 4 in the preparation of a medicament for treating pain caused by arthralgia.

8. The use according to claim 7, characterized in that: The medicine is used to treat pain and swelling in the joints of the limbs, joint dysfunction, joint cavity effusion or soft tissue diseases after trauma caused by wind, cold, dampness and blood stasis.

9. The use according to claim 7, characterized in that: The medicine is a medicine for treating knee osteoarthritis combined with synovitis.

10. The use according to claim 7, characterized in that: The drug is a drug that reverses cartilage damage in knee osteoarthritis, inhibits TNF signaling pathway, reduces cell apoptosis, and slows down the progression of arthritis.

Images