Traditional Chinese medicine composition for treating rheumatoid arthritis

Abstract

The invention belongs to the technical field of traditional Chinese medicines, and relates to a traditional Chinese medicine composition for treating rheumatoid arthritis. The traditional Chinese medicine composition for treating rheumatoid arthritis comprises an aqueous extract, an alcohol extract and a supercritical extract, the aqueous extract is an aqueous extract of Radix Astragali, white peony root and red sage root; the alcohol extracts are alcohol extracts of tripterygium wilfordii, caulis sinomenii and turmeric; the supercritical extract is a supercritical CO2 extract of radix clematidis and zaocys dhumnade. The traditional Chinese medicine composition further comprises a functional component, and the functional component is composed of 2-methyl-5-nitroimidazole and 3-demethylated colchicine. Water extraction, alcohol extraction and supercritical CO2 extraction technologies are comprehensively applied, functional components are introduced, and the ratio of the functional components to traditional Chinese medicine extracts and a preparation process are optimized, so that the treatment effect of the medicine is further enhanced, and a new choice is provided for treatment of rheumatoid arthritis.

Description

Technical Field

[0002] The present invention belongs to the technical field of traditional Chinese medicine and relates to a traditional Chinese medicine composition for treating rheumatoid arthritis. Background Art

[0003] Rheumatoid Arthritis (RA) is an autoimmune disease mainly characterized by chronic, symmetrical, and polyarticular inflammation, often leading to joint destruction, dysfunction, and systemic complications, seriously affecting the quality of life of patients. Currently, the main drugs for treating RA include non-steroidal anti-inflammatory drugs (NSAIDs), glucocorticoids, and disease-modifying antirheumatic drugs (DMARDs), etc. However, these drugs have many limitations during long-term use, such as gastrointestinal side effects, osteoporosis, increased risk of infection, and high treatment costs, etc.

[0004] Traditional Chinese medicine has accumulated rich experience in treating RA, showing good curative effects and low side effects. In the prior art, various traditional Chinese medicine compounds or monomer components have been reported for the treatment of RA, such as Xanthium sibiricum, Angelica pubescens, Angelica sinensis, etc., which play roles through regulating immune responses, inhibiting the release of inflammatory factors, reducing joint damage, and other ways. However, existing traditional Chinese medicine preparations still have deficiencies in aspects such as preparation processes, extraction efficiency of active ingredients, drug stability, and bioavailability, restricting their clinical application effects.

[0005] Therefore, in the technical field of traditional Chinese medicine, it is particularly important to provide an improved traditional Chinese medicine composition for treating rheumatoid arthritis. Summary of the Invention

[0006] The purpose of the present invention is to provide a traditional Chinese medicine composition for treating rheumatoid arthritis. By comprehensively applying water extraction, alcohol extraction, and supercritical CO2 extraction technologies, the present invention realizes the efficient extraction and enrichment of active ingredients from medicinal materials such as Astragalus membranaceus, Paeonia lactiflora, Salvia miltiorrhiza, Tripterygium wilfordii, Sinomenium acutum, Curcuma longa, Clematis chinensis, and Zaocys dhumnades, significantly improving the therapeutic effect of the drug. At the same time, by introducing functional components 2-methyl-5-nitroimidazole and 3-demethylcolchicine, and optimizing their ratios with the traditional Chinese medicine extract and the preparation process, the therapeutic effect of the drug is further enhanced, thus providing a new option for the treatment of rheumatoid arthritis.

[0007] To achieve the above purpose, the technical solution of the present invention is as follows: In the first aspect, the present invention provides a traditional Chinese medicine composition for treating rheumatoid arthritis, and the traditional Chinese medicine composition includes a water extract, an alcohol extract, and a supercritical extract, wherein, The water extract is the water extract of Astragalus membranaceus, Paeonia lactiflora, and Salvia miltiorrhiza; The alcohol extract is the alcohol extract of Tripterygium wilfordii, Sinomenium acutum, and Curcuma longa; The supercritical extract is a supercritical CO2 extract of Clematis chinensis Osbeck and Zaocys dhumnades (Cantor).

[0008] Preferably, the traditional Chinese medicine composition further comprises functional components, which are composed of 2-methyl-5-nitroimidazole and 3-demethylcolchicine, and the mass ratio of 2-methyl-5-nitroimidazole to 3-demethylcolchicine is 5:1.

[0009] Preferably, by weight parts, the traditional Chinese medicine composition comprises 60-70 parts of water extract, 20-30 parts of alcohol extract and 5-10 parts of supercritical extract.

[0010] Preferably, by weight parts, the traditional Chinese medicine composition further comprises 0.1-0.6 part of functional components.

[0011] Preferably, the preparation method of the water extract is as follows: Astragalus membranaceus (Fisch.) Bunge, Paeonia lactiflora Pall. and Salvia miltiorrhiza Bunge are decocted twice according to the mass ratio of 3:2:1, the filtrates are combined and concentrated under reduced pressure to a density of 1.10-1.15 g / cm 3 , and obtained by spray drying; Specifically, the preparation method of the water extract is as follows: Astragalus membranaceus (Fisch.) Bunge, Paeonia lactiflora Pall. and Salvia miltiorrhiza Bunge are put into a multi-functional extraction tank according to the mass ratio of 3:2:1, 10 times the amount of water (w / v) is added, and it is soaked for 30 min; heated to boiling, maintained for 2 h, and filtered to obtain the first extraction liquid; 8 times the amount of water (w / v) is added to the medicinal residues, decocted again for 1.5 h, and the two filtrates are filtered and combined; the filtrate is concentrated under reduced pressure at 80 °C to a density of 1.10-1.15 g / cm 3 , and obtained by spray drying (inlet air temperature 180 °C, outlet air temperature 90 °C, adding 2% maltodextrin as a drying aid); The preparation method of the alcohol extract is as follows: Tripterygium wilfordii Hook. f., Sinomenium acutum (Thunb.) Rehd. et Wils. and Curcuma longa L. are extracted with alcohol twice according to the mass ratio of 1:2:1, the filtrates are combined and obtained by concentration under reduced pressure, centrifugation and drying; Specifically, the preparation method of the alcohol extract is as follows: Tripterygium wilfordii Hook. f. (needs to be processed to reduce toxicity), Sinomenium acutum (Thunb.) Rehd. et Wils. and Curcuma longa L. are put into a reflux extraction tank according to the mass ratio of 1:2:1, 8 times the amount of 60% (v / v) ethanol is added, heated to 78 °C and refluxed for 2 h, and filtered to obtain the first alcohol extraction liquid; 6 times the amount of 60% (v / v) ethanol is added to the medicinal residues, refluxed for 2 h again, and the two alcohol extraction liquids are combined; the alcohol extraction liquid is concentrated under reduced pressure at 50 °C until there is no alcohol smell, centrifuged at 4000 rpm for 15 min to remove insoluble substances, vacuum dried at 60 °C until the water content ≤ 5%, and pulverized through an 80-mesh sieve to obtain; The preparation method of the supercritical extract is as follows: crush Clematis chinensis Osbeck to 40-60 mesh and dry it until the water content ≤ 8%; after processing Zaocys dhumnades, ultrafinely crush it to 80-100 mesh; mix the processed Clematis chinensis Osbeck and Zaocys dhumnades in a mass ratio of 1:1 - 3:1, place them in a supercritical CO2 extraction kettle for supercritical extraction, then conduct primary separation and secondary separation, combine the products of primary separation and secondary separation, and remove the residual entrainer to obtain the supercritical CO2 extract.

[0012] Preferably, the conditions for supercritical extraction are as follows: the CO2 pressure is 20 - 30 MPa, the CO2 flow rate is 20 - 35 kg / h, the temperature is 40 - 50 °C, the time is 1.5 - 3 h, and the entrainer is 3 - 8% (v / v) absolute ethanol; The conditions for primary separation are: the pressure is 5 - 8 MPa and the temperature is 30 - 40 °C; The conditions for secondary separation are: the pressure is 3 - 5 MPa and the temperature is 25 - 35 °C; In the second aspect, a preparation method of the traditional Chinese medicine composition of the present invention is provided. The preparation method is as follows: dissolve the functional components in 2 - 12 mL of 75% ethanol, adsorb them with 2 - 12 g of microcrystalline cellulose in a fluidized bed by spraying (nozzle diameter 0.8 mm) to obtain drug-loaded microspheres, then co-grind them with the supercritical extract (ball mill, 200 rpm × 15 min), then mix them with the water extract and alcohol extract, then conduct wet granulation (the binder is 5% PVP-K30 ethanol solution), pass through a 20-mesh sieve, and finally conduct drying (dry in a fluidized bed at 50 °C until the water content ≤ 5%), and then fill them into enteric-coated capsules, with each capsule containing 0.45 g of medicinal powder.

[0013] In the third aspect, a pharmaceutical preparation is provided, which is made of the traditional Chinese medicine composition of the present invention and pharmaceutically acceptable excipients.

[0014] In the fourth aspect, an application of the traditional Chinese medicine composition of the present invention in the preparation of a pharmaceutical preparation is provided.

[0015] In the fifth aspect, an application of the traditional Chinese medicine composition of the present invention in the preparation of a pharmaceutical preparation for reducing the joint swelling index and / or the incidence rate of abnormal liver function is provided.

[0016] In the sixth aspect, an application of the traditional Chinese medicine composition of the present invention in the preparation of a pharmaceutical preparation for treating an organism with insufficient response to DMARDs is provided.

[0017] In the present invention, the raw materials / traditional Chinese medicines used have the following efficacy / pharmacological effects: Astragalus membranaceus (Fisch.) Bunge: sweet, slightly warm; attributing to the spleen and lung meridians. Tonifying qi and lifting yang, securing the exterior and stopping sweating, promoting diuresis and alleviating edema, inducing the subsidence of swelling and promoting granulation. Indicating spleen deficiency with weakness, middle qi sinking, spontaneous sweating due to exterior deficiency, edema, and difficult or non-healing ulcers.

[0018] White Peony Root: Bitter, sour, slightly cold; acts on the liver and spleen meridians. Nourishes blood and regulates menstruation, astringes yin and stops sweating, soothes the liver and relieves pain, suppresses hyperactive liver-yang. Indicated for sallow complexion due to blood deficiency, irregular menstruation, spontaneous sweating and night sweating, hypochondriac pain and abdominal pain, spasms and pain of the four limbs.

[0019] Salvia Miltiorrhiza: Bitter, slightly cold; acts on the heart and liver meridians. Promotes blood circulation to remove stasis, dredges meridians and relieves pain, clears the heart and removes vexation, cools blood and eliminates carbuncles. Mainly treats chest impediment pain, irregular menstruation, traumatic injury, restlessness and insomnia.

[0020] Thunder God Vine: Bitter, pungent, cool; highly toxic; acts on the liver and kidney meridians. Expels wind and dampness, activates blood circulation and dredges collaterals, reduces swelling and alleviates pain, kills insects and detoxifies.

[0021] Sinomenium Acutum: Bitter, pungent, flat; acts on the liver and spleen meridians. Expels wind-dampness, dredges collaterals, promotes urination.

[0022] Turmeric: Pungent, bitter, warm; acts on the spleen and liver meridians. Breaks blood and promotes qi movement, dredges meridians and relieves pain. Used for chest and hypochondriac stabbing pain, amenorrhea, mass in the abdomen, traumatic injury and swelling pain.

[0023] Clematis Chinensis: Pungent, salty, warm; acts on the bladder meridian. Dredges collaterals and relieves pain, eliminates obstruction of bones. Mainly treats wind-damp arthralgia, limb numbness, fish bone stuck in the throat.

[0024] Zaocys: Sweet, flat; acts on the liver meridian. Expels wind, dredges collaterals, stops convulsions. Used for hemiplegia due to stroke, tetanus, skin pruritus.

[0025] Compared with the prior art, the beneficial effects of the present invention are as follows: (1) Through the synergy of water extraction (Astragalus membranaceus, White Peony Root, Salvia Miltiorrhiza), alcohol extraction (Thunder God Vine, Sinomenium Acutum, Turmeric) and supercritical CO2 extraction (Clematis Chinensis, Zaocys), the present invention efficiently extracts and enriches water-soluble polysaccharides, lipophilic terpenoids and thermosensitive alkaloids respectively. The combined use of the three extraction methods ensures the comprehensive retention and efficient utilization of various active ingredients in the drug.

[0026] (2) The aqueous extract (astragalus polysaccharide, paeoniflorin, tanshinone) reduces the release of pro-inflammatory factors by inhibiting the NF-κB pathway, and at the same time regulates the Th17 / Treg cell balance; the ethanol extract (triptolide, sinomenine, curcumin) targets the JAK-STAT signaling pathway and blocks the activation of synovial fibroblasts; the supercritical extract (clematis saponin, zaocys polypeptide) inhibits the expression of MMP-3 / 9 and slows down the degradation of cartilage matrix. The treatment mechanism of the present invention can be targeted at the complex pathological features of rheumatoid arthritis "immune disorder - inflammation activation - joint destruction", and can reduce the joint swelling index and the incidence of abnormal liver function. Specifically, on the one hand, the aqueous extract inhibits the NF-κB pathway (upstream regulation) → the ethanol extract blocks the JAK-STAT signal (midstream intervention) → the supercritical extract inhibits MMP (downstream protection), which can form a three-level anti-inflammatory network; on the other hand, the aqueous extract regulates the Th17 / Treg balance and the supercritical extract protects the cartilage matrix synergistically, which can delay the pathological progression of RA.

[0027] (3) The present invention significantly enhances the efficacy of the drug by introducing 2-methyl-5-nitroimidazole and 3-demethylcolchicine, and by optimizing their ratios with the traditional Chinese medicine extract and the preparation process. These two functional components synergistically interact with the active components in the traditional Chinese medicine extract and act on the pathogenesis of RA together, achieving a comprehensive treatment of the disease. Specifically, 2-methyl-5-nitroimidazole and 3-demethylcolchicine intervene at multiple links from the initiation (IL-17 signal), amplification (cell migration) to the terminal effect (bone destruction) of the inflammatory cascade, not only significantly relieving the inflammatory symptoms of RA (reducing the joint swelling index and the incidence of abnormal liver function), but also achieving disease-modifying effects through immunomodulation and cell cycle regulation. This dual-target approach can provide a new direction for the treatment of RA, especially suitable for patients who are insufficiently responsive to traditional DMARDs.

[0028] (4) The present invention improves the stability and bioavailability of the drug by optimizing the preparation process. Advanced drying techniques such as spray drying and vacuum drying are used to effectively retain the active components in the drug; preparation techniques such as fluidized bed spray adsorption and co-grinding are used to improve the dispersibility and uniformity of the drug; the sustained release and targeted delivery of functional components are achieved through the fluidized bed spray adsorption technique of microcrystalline cellulose drug-loaded microspheres; at the same time, a dense matrix structure is formed by wet granulation + enteric coating, which can delay the drug release and improve the storage stability. Detailed implementation manners

[0029] The following will specifically describe the present invention in combination with the detailed implementation manners and examples, and the advantages and various effects of the present invention will be presented more clearly therefrom. Those skilled in the art should understand that these detailed implementation manners and examples are used to illustrate the present invention, rather than to limit the present invention.

[0030] Next, the technical solution of the present invention will be described in conjunction with embodiments. However, the present invention is not limited to the following embodiments. The experimental methods and detection methods described in each embodiment are all conventional methods unless otherwise specified; the reagents and materials described are all commercially available unless otherwise specified.

[0031] 2-Methyl-5-nitroimidazole was purchased from Wuhan Lana White Pharmaceutical Chemical Co., Ltd., with a CAS number of 696-23-1 and an active ingredient content of 98%.

[0032] 3-Demethylcolchicine was purchased from Sichuan Jingcui Tiancheng Pharmaceutical Technology Co., Ltd., with a CAS number of 7336-33-6 and HPLC≥98%.

[0033] Example 1 This example provides a traditional Chinese medicine composition for treating rheumatoid arthritis. By weight, the traditional Chinese medicine composition includes 60 g of water extract, 20 g of alcohol extract, 5 g of supercritical extract, and 0.1 g of functional ingredient; The water extract is the water extract of Astragalus membranaceus, Paeonia lactiflora, and Salvia miltiorrhiza; The alcohol extract is the alcohol extract of Tripterygium wilfordii, Sinomenium acutum, and Curcuma longa; The supercritical extract is the supercritical CO2 extract of Clematis chinensis and Zaocys dhumnades.

[0034] The functional ingredient is composed of 2-methyl-5-nitroimidazole and 3-demethylcolchicine, and the mass ratio of 2-methyl-5-nitroimidazole to 3-demethylcolchicine is 5:1.

[0035] Example 2 This example provides a traditional Chinese medicine composition for treating rheumatoid arthritis. By weight, the traditional Chinese medicine composition includes 70 g of water extract, 30 g of alcohol extract, 10 g of supercritical extract, and 0.6 g of functional ingredient; The water extract is the water extract of Astragalus membranaceus, Paeonia lactiflora, and Salvia miltiorrhiza; The alcohol extract is the alcohol extract of Tripterygium wilfordii, Sinomenium acutum, and Curcuma longa; The supercritical extract is the supercritical CO2 extract of Clematis chinensis and Zaocys dhumnades.

[0036] The functional ingredient is composed of 2-methyl-5-nitroimidazole and 3-demethylcolchicine, and the mass ratio of 2-methyl-5-nitroimidazole to 3-demethylcolchicine is 5:1.

[0037] Example 3 This example provides a traditional Chinese medicine composition for treating rheumatoid arthritis. By weight, the traditional Chinese medicine composition includes 60 g of water extract, 20 g of alcohol extract, 10 g of supercritical extract, and 0.6 g of functional ingredient; The aqueous extract is the aqueous extract of Astragalus membranaceus, Paeonia lactiflora, and Salvia miltiorrhiza; The ethanol extract is the ethanol extract of Tripterygium wilfordii, Sinomenium acutum, and Curcuma longa; The supercritical extract is the supercritical CO2 extract of Clematis chinensis and Zaocys dhumnades.

[0038] The functional components are composed of 2-methyl-5-nitroimidazole and 3-demethylcolchicine, and the mass ratio of 2-methyl-5-nitroimidazole to 3-demethylcolchicine is 5:1.

[0039] Example 4 This example provides a preparation method of the traditional Chinese medicine composition. The preparation method is as follows: Dissolve the functional components in 2 - 12 mL of 75% ethanol, and adsorb them with 2 - 12 g of microcrystalline cellulose in a fluidized bed by spraying (nozzle diameter 0.8 mm) to obtain drug-loaded microspheres. Then, co-grind them with the supercritical extract (ball mill, 200 rpm × 15 min). Next, mix them with the aqueous extract and the ethanol extract, and then perform wet granulation (the binder is a 5% PVP-K30 ethanol solution) through a 20-mesh sieve. Finally, dry them (fluidized bed drying at 50°C until the water content ≤ 5%), and fill them into enteric-coated capsules, with each capsule containing 0.45 g of the drug powder. Among them, The preparation method of the aqueous extract is as follows: Put Astragalus membranaceus, Paeonia lactiflora, and Salvia miltiorrhiza into a multi-functional extraction tank according to a mass ratio of 3:2:1, add 10 times the amount of water (w / v), and soak for 30 min; heat to boiling and maintain for 2 h, then filter to obtain the first extraction liquid; add 8 times the amount of water (w / v) to the residue, decoct again for 1.5 h, and filter and combine the two filtrates; the filtrate is concentrated under reduced pressure at 80°C to a density of 1.10 - 1.15 g / cm 3 , and obtain it by spray drying (inlet air temperature 180°C, outlet air temperature 90°C, adding 2% maltodextrin as a drying aid); The preparation method of the ethanol extract is as follows: Put Tripterygium wilfordii (which needs to be processed to reduce toxicity), Sinomenium acutum, and Curcuma longa into a reflux extraction tank according to a mass ratio of 1:2:1, add 8 times the amount of 60% (v / v) ethanol, heat to 78°C and reflux for 2 h, then filter to obtain the first ethanol extraction liquid; add 6 times the amount of 60% (v / v) ethanol to the residue, repeat refluxing for 2 h, and combine the two ethanol extraction liquids; the ethanol extraction liquid is concentrated under reduced pressure at 50°C until there is no alcohol smell, centrifuged at 4000 rpm for 15 min to remove insoluble substances, and vacuum dried at 60°C until the water content ≤ 5%, and then pulverized through an 80-mesh sieve to obtain it; The preparation method of the supercritical extract is as follows: Pulverize Clematis chinensis to 40 - 60 mesh and dry it until the water content ≤ 8%; ultra-finely pulverize Zaocys dhumnades to 80 - 100 mesh after processing; mix Clematis chinensis and Zaocys dhumnades according to a mass ratio of 1:1 - 3:1, place them in a supercritical CO2 extraction kettle for supercritical extraction, then perform primary separation and secondary separation, combine the primary separation and secondary separation products, and remove the residual entrainer to obtain the supercritical CO2 extract; The conditions for supercritical extraction are as follows: the CO2 pressure is 20 - 30 MPa, the CO2 flow rate is 20 - 35 kg / h, the temperature is 40 - 50 °C, the time is 1.5 - 3 h, and the entrainer is 3 - 8% (v / v) absolute ethanol; The conditions for primary separation are: the pressure is 5 - 8 MPa and the temperature is 30 - 40 °C; The conditions for secondary separation are: the pressure is 3 - 5 MPa and the temperature is 25 - 35 °C.

[0040] Comparative Example 1 This comparative example is the same as Example 3, except that: Paeonia lactiflora Pall. is replaced by Paeonia veitchii Lynch.

[0041] Comparative Example 2 This comparative example is the same as Example 3, except that: the supercritical CO2 extracts of Clematis chinensis Osbeck and Zaocys dhumnades (Cantor) are replaced by: Clematis chinensis Osbeck and Zaocys dhumnades (Cantor) are mixed in a mass ratio of 1:1 - 3:1 and refluxed with ethanol for extraction, filtration, concentration, and drying.

[0042] Comparative Example 3 This comparative example is the same as Example 3, except that: 2 - methyl - 5 - nitroimidazole is not added.

[0043] Effect verification The experimental animals and the method of establishing the model are as follows: 1. Experimental animals SPF - grade female SD rats (8 - month - old, body weight 350 ± 10 g), adaptively fed for 1 week, freely fed and watered, room temperature 22 ± 2 °C, relative humidity 50 ± 10%, 12 - h light - dark cycle.

[0044] 2. Establishment of rheumatoid arthritis (CIA) model (1) Preparation of collagen emulsion: Dissolve bovine type II collagen (Sigma, C9301) in 0.1 mol / L acetic acid (pH 3.0) to prepare a 2 mg / mL collagen solution, stir overnight at 4 °C; mix the collagen solution and Freund's complete adjuvant (Sigma, F5881) in a volume ratio of 1:1, and repeatedly aspirate with a syringe until emulsification is complete (the emulsion droplets form complete spheres when dropped into water and do not spread within 30 min).

[0045] (2) Immunization for model establishment: Primary immunization: Disinfect the skin at the root of the rat tail, back, and right hind paw toes (75% alcohol), and intradermally inject 0.4 mL / rat of collagen emulsion (3 points in total, 0.13 mL per point) to form a skin papule with a diameter ≥ 3 mm.

[0046] Booster immunization: Inject once in the same way on the 7th day.

[0047] Criteria for successful model establishment: Paw volume: Measured by the drainage method, ≥0.80 mL (0.50 ± 0.05 mL in the normal group).

[0048] Ankle joint diameter: Measured with a vernier caliper, an increase of ≥12 mm (6.5 ± 0.5 mm in the normal group).

[0049] Joint swelling index: ≥3 points (scoring criteria are shown in Table 1 below).

[0050] (3)Adaptive swallowing treatment Use size 0 enteric-coated capsules (sized for rat swallowing), filled with medicinal powder (0.45 g per capsule), and train rats to adapt to capsule swallowing (give empty capsules daily for 1 week in advance, supplemented with a reward mechanism); use a special capsule feeder to send the enteric-coated capsule to the pharynx of the rat to prompt natural swallowing; randomly select 3 rats, orally administer barium sulfate-labeled capsules, and confirm by X-ray imaging that the capsules have completely entered the intestine.

[0051] 3. Verification of enteric characteristics Take 6 batches of enteric-coated capsules (3 capsules per batch), simulate the gastrointestinal fluid conditions (gastric fluid pH 1.2, intestinal fluid pH 6.8), sample at 1 h, 2 h, and 4 h respectively, and detect the release rate by HPLC. The results are expressed as mean ± standard deviation (n = 6), as shown in Table 2 specifically. It can be seen from Table 2 that under the simulated gastrointestinal fluid conditions, the drug is slowly released in the stomach and can be rapidly released in the intestine for this enteric-coated capsule, meeting the design requirements of the enteric-coated capsule, having good gastric dissolution and intestinal release characteristics, and being able to effectively release the drug in the intestine.

[0052] 4. Grouping and exclusion Grouping: Exclude individuals with failed modeling (paw volume increase < 0.80 mL), and randomly divide 90 rats into 9 groups (10 rats per group).

[0053] Blank group: Normal rats, without modeling.

[0054] Model group: Induce CIA model, and intragastrically administer normal saline.

[0055] Positive group: Methotrexate (positive drug).

[0056] Experimental groups: Experimental group 1 is the traditional Chinese medicine composition prepared according to Example 4 of Example 1, experimental group 2 is the traditional Chinese medicine composition prepared according to Example 4 of Example 2, and experimental group 3 is the traditional Chinese medicine composition prepared according to Example 4 of Example 3.

[0057] Control groups: Control group 1 is the traditional Chinese medicine composition prepared according to Example 4 of Comparative Example 1, control group 2 is the traditional Chinese medicine composition prepared according to Example 4 of Comparative Example 2, and control group 3 is the traditional Chinese medicine composition prepared according to Example 4 of Comparative Example 3.

[0058] 5. Evaluation of joint swelling index and paw volume On the 28th day, the joint swelling index and paw volume were evaluated. The specific method was as follows: The rats were placed in a transparent observation box and kept in a natural standing position. Two researchers independently performed blind scoring on the bilateral ankle joints and wrist joints and took the average value (n = 10). Scoring was performed at a fixed time every week (such as 9 - 10 am on Monday every week) to avoid the interference of circadian rhythm. The results are shown in Table 3.

[0059] As can be seen from Table 3, compared with Example 1, Example 2, and Example 3, although the joint swelling index and paw volume of the positive group were improved, they were not as improved as those of Examples 1 - 3 of the present invention, especially not as significant as the effect of Example 3. The joint swelling index of Example 3 was the lowest, and the paw volume was also close to the level of the blank group, indicating that the treatment method of Example 3 had the best effect on improving joint swelling and increasing paw volume. The joint swelling index and paw volume of Comparative Example 1, Comparative Example 2, and Comparative Example 3 were all higher than those of the example group but lower than those of the model group, indicating that these comparison methods could also improve joint swelling and increasing paw volume to a certain extent, but the effect was not as significant as that of the experimental group.

[0060] 6. Detection of liver function After evaluating the joint swelling index and paw volume, the rats were sacrificed. The rats were fasted for 12 h before sacrifice and allowed free access to water. After collecting blood from the eyeballs, the rats were sacrificed by cervical dislocation, and the livers were collected for liver function (ALT, AST) detection. Among them, the instruments for ALT and AST detection were fully automatic biochemical analyzers (Hitachi 7180), and the reagent kits for ALT and AST detection were purchased from Zhong Sheng Beikong Biotech Co., Ltd. (product numbers C009 - 2 and C010 - 2), and the operation was carried out strictly according to the instructions. The results are shown in Table 4. As can be seen from Table 4, the ALT and AST levels of Examples 1, 2, and 3 were all lower than those of the model group and the positive group, and were close to or reached the level of the blank group, indicating that the experimental group had a certain effect on improving liver injury. Among them, the ALT and AST levels of Example 3 were the lowest and the effect was the best. The ALT and AST levels of Comparative Example 1, 2, and 3 were all lower than those of the model group but higher than those of the experimental group, indicating that these comparison groups had a worse effect on improving liver injury than the experimental group.

[0061] Table 1 Scoring criteria for joint swelling index Table 2 Enteric characteristics Table 3 Joint swelling index and paw volume Table 4 Results of liver function detection It should be noted that the traditional Chinese medicine composition of the present invention synergistically combines multi-dimensional active ingredients of water extract, alcohol extract and supercritical extract to cover three major pathways of inflammation inhibition, immune regulation and liver protection. Among them, astragalus polysaccharide and paeoniflorin in the water extract can target and inhibit NF-κB signal transduction, triptolide in the alcohol extract blocks abnormal immune activation through the JAK-STAT pathway, and clematis saponin in the supercritical extract promotes the differentiation of Treg cells to reconstruct immune homeostasis. In particular, the functional components 2-methyl-5-nitroimidazole and 3-demethylcolchicine form a complex in a mass ratio of 5:1, which can synergistically antagonize the synovial destruction mediated by IL-17A, and at the same time reduce the metabolic toxicity of methotrexate by competitively inhibiting the hepatic enzyme CYP3A4. The composition of the present invention adopts an enteric capsule delivery system, which can improve bioavailability and reduce gastrointestinal irritation, and is suitable for rheumatoid arthritis patients with insufficient response to long-term DMARDs treatment.

[0062] To further illustrate the beneficial effects of the present invention, the following comparative examples are provided: Astragalus membranaceus, Salvia miltiorrhiza, Tripterygium wilfordii, Sinomenium acutum, Curcuma longa, Clematis chinensis, Zaocys dhumnades, 2-methyl-5-nitroimidazole and 3-demethylcolchicine were respectively replaced with other substances of similar properties, and the remaining conditions were the same as in Example 3. The effects of treating rheumatoid arthritis were verified according to the method in the effect verification, and the results were similar to those in Comparative Example 1 above.

[0063] To further illustrate the beneficial effects of the present invention, the following comparative examples are provided: The mass ratio of 2-methyl-5-nitroimidazole and 3-demethylcolchicine was adjusted to 4:1 or 5:2, and the remaining conditions were the same as in Example 3. The effects of treating rheumatoid arthritis were verified according to the method in the effect verification, and the results were similar to those in Comparative Example 2 above.

[0064] To further illustrate the beneficial effects of the present invention, the following comparative examples are provided: 3-demethylcolchicine, Astragalus membranaceus, Paeonia lactiflora, Salvia miltiorrhiza, Tripterygium wilfordii, Sinomenium acutum, Curcuma longa, Clematis chinensis and Zaocys dhumnades were not added respectively, and the remaining conditions were the same as in Example 3. The effects of treating rheumatoid arthritis were verified according to the method in the effect verification, and the results were similar to those in Comparative Example 3 above.

[0065] It can be seen from the effect verification test that the formula system of the traditional Chinese medicine composition for treating rheumatoid arthritis provided by the present invention is an integral whole. Specifically, each raw material and each method step in the traditional Chinese medicine composition for treating rheumatoid arthritis support each other functionally and have an interaction relationship. It is precisely because of the functional mutual support and interaction relationship between each raw material, each method step, and between the method step and the raw material that the traditional Chinese medicine composition of the present invention's formula system can target the complex pathological characteristics of "immune disorder - inflammation activation - joint destruction" of rheumatoid arthritis, and can reduce the joint swelling index and the incidence of abnormal liver function.

[0066] In addition, the embodiments of the present invention also verified the stability, and it was confirmed that after 6 months of accelerated test, the content change rate of the experimental groups (Example 1, Example 2, Example 3) < 5%, which was significantly better than that of the control groups (Comparative Example 1, Comparative Example 2, Comparative Example 3).

[0067] It should be understood that the present invention disclosed is not limited to the specific methods, schemes and substances described, as these can vary. It should also be understood that the terms used herein are for the purpose of describing specific embodiments only and are not intended to limit the scope of the present invention, the scope of which is only limited by the appended claims.

Claims

1. A traditional Chinese medicine composition for treating rheumatoid arthritis, characterized in that, The traditional Chinese medicine composition comprises an aqueous extract, an ethanol extract and a supercritical extract, wherein, the aqueous extract is the aqueous extract of Astragalus membranaceus, Paeonia lactiflora and Salvia miltiorrhiza; the ethanol extract is the ethanol extract of Tripterygium wilfordii, Sinomenium acutum and Curcuma longa; the supercritical extract is the supercritical CO2 extract of Clematis chinensis and Zaocys dhumnades.

2. The traditional Chinese medicine composition according to claim 1, characterized in that, The traditional Chinese medicine composition further comprises functional components, which are composed of 2-methyl-5-nitroimidazole and 3-demethylcolchicine, and the mass ratio of 2-methyl-5-nitroimidazole to 3-demethylcolchicine is 5:

1.

3. The traditional Chinese medicine composition according to claim 1, wherein Calculated by weight parts, the traditional Chinese medicine composition comprises 60-70 parts of aqueous extract, 20-30 parts of ethanol extract and 5-10 parts of supercritical extract.

4. The traditional Chinese medicine composition according to claim 2, wherein Calculated by weight parts, the traditional Chinese medicine composition further comprises 0.1-0.6 part of functional components.

5. The traditional Chinese medicine composition according to claim 1, wherein, The preparation method of the water extract is as follows: Astragalus membranaceus, Paeonia lactiflora Pall., and Salvia miltiorrhiza Bunge are decocted twice in water according to a mass ratio of 3:2:1, and the filtrates are combined and concentrated under reduced pressure to a density of 1.10 - 1.15 g / cm 3 , and then obtained by spray drying; the preparation method of the ethanol extract is: Tripterygium wilfordii, Sinomenium acutum and Curcuma longa are subjected to ethanol extraction twice according to the mass ratio of 1:2:1, the filtrates are combined and concentrated under reduced pressure, centrifuged and dried to obtain; the preparation method of the supercritical extract is: Clematis chinensis is crushed to 40-60 meshes and dried to a water content of ≤8%; Zaocys dhumnades is superfine crushed to 80-100 meshes after being processed; Clematis chinensis and Zaocys dhumnades are mixed according to the mass ratio of 1:1-3:1, placed in a supercritical CO2 extraction kettle for supercritical extraction, followed by primary separation and secondary separation, the products of primary separation and secondary separation are combined, and the residual entrainer is removed to obtain the supercritical CO2 extract.

6. The traditional Chinese medicine composition according to claim 5, wherein, the conditions of the supercritical extraction are: the CO2 pressure is 20-30 MPa, the CO2 flow rate is 20-35 kg / h, the temperature is 40-50 °C, the time is 1.5-3 h, and the entrainer is 3-8% absolute ethanol; the conditions of the primary separation are: the pressure is 5-8 MPa and the temperature is 30-40 °C; the conditions of the secondary separation are: the pressure is 3-5 MPa and the temperature is 25-35 °C.

7. A method for preparing the traditional Chinese medicine composition according to any one of claims 1-6, characterized in that, The preparation method is: the functional components are dissolved in 2-12 mL of 75% ethanol, spray adsorbed with 2-12 g of microcrystalline cellulose in a fluidized bed to obtain drug-loaded microspheres, co-ground with the supercritical extract, then mixed with the aqueous extract and the ethanol extract, and finally granulated, dried and filled into enteric capsules.

8. A pharmaceutical preparation, characterized in that, It is made from the traditional Chinese medicine composition according to any one of claims 1-6 and pharmaceutically acceptable excipients.

9. Use of the traditional Chinese medicine composition according to any one of claims 1-6 in the preparation of a pharmaceutical preparation for reducing the joint swelling index and / or the incidence rate of abnormal liver function.

10. Use of the traditional Chinese medicine composition according to any one of claims 1-6 in the preparation of a pharmaceutical preparation for treating a body with insufficient response to DMARDs.

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