Compound preparation for treating rheumatoid arthritis and preparation method thereof

Abstract

The invention relates to the technical field of traditional Chinese medicine, and particularly discloses a compound preparation for treating rheumatoid arthritis and a preparation method thereof.The preparation is prepared from salvia miltiorrhiza, rhodiola rosea, semen cuscutae, asparagus cochinchinensis, fritillaria pallidiflora, curcuma zedoary and radix clematidis through solvent extraction, impurity removal, concentration or drying. The traditional Chinese medicine composition and medically acceptable auxiliary materials are prepared into granules, oral preparations, pills, capsules, tablets and the like. The compound preparation has the effects of expelling wind, removing cold, removing dampness, tonifying qi, nourishing blood, nourishing liver-kidney deficiency, promoting blood circulation, removing blood stasis, diminishing swelling, eliminating phlegm, dredging collaterals and relieving pain. The traditional Chinese medicine composition is used for treating rheumatoid arthritis, ankylosing spondylitis, chronic arthritis and rheumatic sciatica. The compound preparation disclosed by the invention can be used for effectively relieving joint tissue injury and inflammation of a Fisher adjuvant induced arthritis mouse, inhibiting the activation of DC and macrophages in a fossa lymph node and a spleen and the secretion of IL-6 and TNF-a, reducing the number of Th1 and Th17, increasing the number of Treg cells, increasing the proportion of Th2 / Th1 cells and recovering the immune balance of CD4 cells, and has a treatment effect.

Description

Technical Field

[0001] The present invention belongs to the technical field of traditional Chinese medicine, and in particular relates to a compound preparation for treating rheumatoid arthritis and a preparation method thereof. Background Art

[0002] Rheumatoid arthritis (RA) is an autoimmune disease characterized by polyarticular inflammation, including synovial tissue proliferation, inflammation, and cartilage destruction. RA is a recurrent, persistent disease with a high disability rate that can affect multiple body systems, making it a major health threat.

[0003] RA is caused by a combination of factors, and its specific pathogenesis remains incompletely understood. With the rapid advancement of immunology in recent years, the immunological etiology of RA is gradually being elucidated. RA is a type of disease in which antigen-presenting cells (such as dendritic cells, DCs) present pathological autoantigens, activating autoreactive T and B cells, leading to immune dysfunction and multi-tissue and organ damage, with CD4+ T cells playing a key role. Currently, Western medicine offers a variety of treatments for this disease, but drug therapy remains an indispensable tool for treating rheumatoid arthritis. Western medicine is the primary treatment, including nonsteroidal anti-inflammatory drugs (NSAIDs), disease-modifying antirheumatic drugs (DMARDs), glucocorticoids (GCs), and biologics. These drugs are rapidly effective and reliably effective, but they can develop varying degrees of drug resistance and side effects with prolonged and frequent use. Furthermore, their effectiveness is insufficient to meet the needs of the growing number of patients, limiting their clinical application. At present, natural therapies and green therapies that return to nature have been widely recognized by people, and people in many countries around the world are becoming more and more accepting of my country's natural medicines and traditional therapies.

[0004] Given the high incidence of RA, searching for natural remedies and extracting their active ingredients has become an effective approach for modern medicine to explore and develop new drugs. Therefore, developing anti-RA drugs from traditional Chinese medicines and ethnic medicines has important social and economic value.

[0005] Traditional Chinese medicine (TCM) currently offers a rich and diverse range of treatments, including anti-inflammatory, analgesic, and symptom relief, as well as improved microcirculation and immune regulation. It can also enhance efficacy and reduce toxicity, alleviating some of the adverse reactions of Western medications. This has significantly enhanced the treatment of RA. By combining some of TCM's distinctive therapies with traditional treatments, TCM can accelerate recovery and achieve better clinical outcomes.

[0006] In recent years, Traditional Chinese Medicine (TCM) has attracted significant attention for its multi-pathway, multi-target mechanisms of action, as well as its low toxicity and safety as a natural product. Utilizing TCM resources to regulate a patient's immune balance is a key area of ​​focus for the future treatment of autoimmune diseases. Traditional Chinese Medicine (TCM) boasts a unique medical theoretical framework and numerous classic formulas. Its classic formulas and therapeutic efficacy, in particular, for the treatment of immune diseases, have garnered significant attention and recognition within the medical community. Over centuries of development, TCM has continuously matured its diagnostic model for RA and developed unique, specialized formulas and treatments. TCM also boasts a diverse range of ingredients, is less susceptible to drug resistance, and offers minimal side effects. Furthermore, TCM can be tailored to the specific patient's condition, resulting in distinctive treatment options, definite efficacy, and a low risk of relapse. This makes TCM a powerful tool for treating patients' conditions and maintaining their health, opening up new avenues for the treatment of this disease. Summary of the Invention

[0007] The object of the present invention is to provide a compound preparation for treating rheumatoid arthritis and a preparation method thereof, so as to solve the problems raised in the above background technology.

[0008] To achieve the above object, the present invention provides the following technical solutions:

[0009] A compound preparation for treating rheumatoid arthritis, comprising the following raw materials in parts by weight: 5-100 parts of salvia miltiorrhiza, 5-200 parts of rhodiola rosea, 5-100 parts of dodder seeds, 5-100 parts of asparagus cochinchinensis, 5-100 parts of fritillaria cirrhosa, 5-100 parts of zedoaria, 5-100 parts of clematis root, and medically acceptable excipients of 1-50 parts of lactose, 1-5 parts of starch, 5-100 parts of sucrose, 1-50 parts of dextrin, and pre- 1-50 parts of gelatinized starch, 5-100 parts of microcrystalline cellulose, 5-100 parts of cross-linked polyvinyl pyrrolidone, 5-100 parts of low-substituted hydroxypropyl cellulose, 5-100 parts of sodium carboxymethyl starch, 1-50 parts of talc, 1-50 parts of micropowdered silica gel, 1-50 parts of magnesium stearate, 5-100 parts of polyethylene glycol, 10-200 parts of honey, 1-50 parts of stevia, and 10-50 parts of sorbic acid. Made into granules, oral preparations, pills, tablets, capsules, syrups, and honey pastes

[0010] Preferably, the compound preparation is used to prepare oral preparations including but not limited to pills, tablets, capsules, syrups, and honey pastes.

[0011] A method for preparing a compound preparation for treating rheumatoid arthritis comprises the following steps:

[0012] S1. Grind 5-100 parts of Salvia miltiorrhiza, 5-200 parts of Rhodiola rosea, 5-100 parts of Cuscuta chinensis, 5-100 parts of Asparagus cochinchinensis, 5-100 parts of Fritillaria cirrhosa, 5-100 parts of Curcuma zedoaria, and 5-100 parts of Clematidis to 10-100 mesh size, mix thoroughly, and extract with 5-25 times the amount of water or 10%-95% ethanol at a temperature of 10-100° C. for 1-4 times, each extraction time being 1-6 hours. Combine the extracts and filter to obtain a water extract or an alcohol extract.

[0013] S2. The water extract or alcohol extract in step a is concentrated under reduced pressure at a temperature of 40-80° C. to an extract with a relative density of 1.10-1.40 to obtain a water extract or an alcohol extract, and the water extract or the alcohol extract is vacuum dried at a temperature of 10-80° C. and crushed to obtain a water extract powder or an alcohol extract powder;

[0014] Alternatively, the water-extracted extract powder or alcohol-extracted extract powder in step b is mixed with 10%-95% ethanol as a binder and auxiliary materials such as sucrose, lactose or starch 10-200 parts and dextrin 10-100 parts, and the mixture is thoroughly mixed and prepared into granules according to conventional pharmaceutical methods.

[0015] Alternatively, the water-extracted extract powder or the alcohol-extracted extract powder in step b is added with purified water, 1-50 parts of stevia, and 1-50 parts of sorbic acid, and the mixture is filtered on a plate and frame, packaged, and sterilized to prepare an oral solution;

[0016] Alternatively, the water-extracted extract powder or the alcohol-extracted extract powder in step b is fully mixed with 10-200 parts of honey and 5-100 parts of polyethylene glycol as auxiliary materials, and the mixture is prepared into pills according to conventional pharmaceutical methods.

[0017] Alternatively, the water-extracted extract powder or alcohol-extracted extract powder in step b is mixed with 1%-10% cross-linked polyvinyl pyrrolidone and 10%-95% ethanol solution as a binder, and excipients including 1-50 parts of lactose, 5-100 parts of microcrystalline cellulose, 5-100 parts of low-substituted hydroxypropyl cellulose, 5-100 parts of sodium carboxymethyl starch, 1-50 parts of pregelatinized starch, 1-50 parts of micropowdered silica gel, 1-50 parts of magnesium stearate, and 1-50 parts of talc, and prepared into tablets or capsules according to conventional pharmaceutical methods;

[0018] Alternatively, the water-extracted extract powder or the alcohol-extracted extract powder in step b is added with purified water, mixed thoroughly with 5-100 parts of sucrose as an auxiliary material, filtered, packaged, sterilized, and prepared into a syrup according to conventional methods;

[0019] Alternatively, the water-extracted extract powder or the alcohol-extracted extract powder in step b is added with 10-200 parts of honey, stirred and mixed, packaged, sterilized, and prepared into a honey paste according to a conventional method.

[0020] A compound preparation for treating rheumatoid arthritis is used to treat rheumatoid arthritis

[0021] Compared with the prior art, the present invention has the following beneficial effects:

[0022] The raw materials of the preparation of the present invention are salvia miltiorrhiza, rhodiola rosea, dodder seed, asparagus cochinchinensis, fritillaria cirrhosa, zedoaria zedoaria, and clematis root. The raw materials are extracted with solvents, removed impurities, concentrated or dried, and then prepared into granules, oral preparations, pills, tablets, capsules, syrups, or honey pastes with medically acceptable excipients. The compound preparation has the effects of dispelling wind, cold, and dampness, replenishing qi and nourishing blood, nourishing liver and kidney deficiency, promoting blood circulation, removing blood stasis, and reducing swelling, and removing phlegm, unblocking meridians, and relieving pain. It is commonly used clinically to treat rheumatoid arthritis, ankylosing spondylitis, chronic arthritis, and rheumatic sciatica. Pharmacodynamic studies have shown that the compound preparation of the present invention effectively alleviates joint tissue damage and inflammation in arthritis mice induced by Freund's adjuvant, inhibits DC and macrophage activation and the secretion of IL-6 and TNF-α in the leg lymph nodes and spleen, reduces the number of Th1 and Th17 cells, increases the number of Treg cells, improves the Th2 / Th1 cell ratio, and restores CD4 cell immune balance.

[0023] At the same time, compared with Western medicine for treating rheumatoid arthritis, the medicament prepared by the present invention has no side effects, no drug resistance and no physical dependence. The preparation process is simple, the cost is low, and it has certain market competitiveness. At the same time, the source of medicinal materials is rich, the quality can be guaranteed, and it is suitable for industrial large-scale production. BRIEF DESCRIPTION OF THE DRAWINGS

[0024] Figure 1 This is a schematic diagram showing the degree of relief of edema in the soles and joints of the feet according to an embodiment of the present invention;

[0025] Figure 2 Schematic diagram of experimental modeling and treatment process in an embodiment of the present invention;

[0026] Figure 3 Schematic diagram of the experimental foot and joint edema in an embodiment of the present invention;

[0027] Figure 4 This is a diagram showing experimental joint tissue damage and HE staining results in an embodiment of the present invention;

[0028] Figure 5 This is a graph of cytokines in experimental serum in an embodiment of the present invention;

[0029] Figure 6 This is a picture of DCs and macrophages in the experimental femoral lymph nodes in the examples of the present invention;

[0030] Figure 7 Graph showing the maturation status and secreted cytokines of DCs and macrophages in mouse spleen in an example of the present invention.

[0031] Figure 8 This is a graph showing the number of CD4+ T cell subtypes in the experiment in the embodiment of the present invention;

[0032] Figure 9 CD in the spleen of experimental mice in the embodiment of the present invention + Expression profile of cytokines secreted by cells. DETAILED DESCRIPTION

[0033] The following will clearly and completely describe the technical solutions in the embodiments of the present invention in conjunction with the accompanying drawings. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without making creative efforts are within the scope of protection of the present invention.

[0034] The compound preparation for treating rheumatoid arthritis of the present invention is guided by traditional Chinese medicine theory, inherits the medication characteristics of traditional Chinese medicine, and adopts modern extraction methods to obtain the active ingredients in the compound medicine based on the long-term folk clinical practice of the Chinese nation, thereby ensuring the content of the active ingredients and improving the bioavailability. The pharmacological properties of the compound preparation are: anti-inflammatory, dispersing and analgesic, and it is used to treat rheumatoid arthritis, ankylosing spondylitis, chronic arthritis, and rheumatic sciatica.

[0035] The preparation method and use of the compound preparation for treating rheumatoid arthritis of the present invention, wherein the functions and indications of each Chinese medicinal ingredient are:

[0036] Danshen: This herb is the dried root and rhizome of Salvia miltiorrhiza Bge., a plant of the Lamiaceae family. It is harvested in spring and autumn, washed, and sun-dried. It promotes blood circulation, removes blood stasis, relieves menstrual pain, clears the heart and relieves restlessness, and cools the blood and eliminates carbuncles. It is used for chest pain, pain caused by heat, restlessness and insomnia, irregular menstruation, dysmenorrhea, and amenorrhea. Take 10-15g internally.

[0037] Rhodiola rosea: This product is the dried root and rhizome of Rhodiola crenulata (Hook.f.et Thoms.) H.Ohba, a plant of the Crassulaceae family. It is harvested in autumn after the flower stems wither, the rough bark removed, washed, and sun-dried. It has the effects of tonifying qi and clearing the lungs, improving intelligence and nourishing the heart, astringing and stopping bleeding, and dispersing blood stasis and reducing swelling. It is used for qi deficiency and weakness, chills after illness, shortness of breath and fatigue, cough due to lung heat, hemoptysis, leucorrhea and diarrhea, and traumatic injuries. It is used to treat impotence, tracheitis, weakness, general fatigue, chest tightness, difficulty breathing, and purple lips and palms. Take 6-12g orally.

[0038] Cuscuta auctralis R.Br or Cuscuta chinensis Lam, both southern plants of the Convolvulaceae family, are harvested in autumn when the fruits are ripe, sun-dried, the seeds removed, and impurities removed. It nourishes the liver and kidneys, strengthens essence and reduces urination, stabilizes pregnancy, improves eyesight, and relieves diarrhea. It can be used externally to dispel wind and remove freckles. It is used for liver and kidney deficiency, soreness of the waist and knees, enuresis and frequent urination, kidney deficiency with fetal bleeding, restless fetuses, diarrhea caused by spleen and kidney deficiency, and external treatment of vitiligo. Take 6-12g internally.

[0039] Asparagus cochinchinensis (Lour) Merr., a plant of the Liliaceae family, is a dried root tuber. It is harvested in autumn and winter, washed, and dried. It nourishes yin and moistens dryness, clears the lungs, and promotes fluid production. It is used for dry coughs caused by lung dryness, sudden coughs with sticky phlegm, soreness of the waist and knees, hot flashes due to bone steaming, internal heat and thirst, loss of body fluid due to febrile illness, dry throat and thirst, constipation caused by dry intestines, and to clear the meridians and relieve asthma. Take 6-12g internally.

[0040] Fritillaria walujewii: Also known as yellow-flowered Fritillaria and Suhuguli, this product is the dried bulb of the Xinjiang Fritillaria (Fritillaria walujewii Regel.) or the Ili Fritillaria (Fritillaria pallidiflora Schrenk.), both of the Liliaceae family. It is harvested in summer and autumn, desilted, sun-dried, and the fibrous roots and outer bark removed. It has the effects of clearing heat and moistening the lungs, resolving phlegm and relieving coughs. It is used for dry coughs caused by lung heat, dry coughs with little phlegm, coughs due to yin deficiency, and coughs with blood in the sputum. Take 3-9g orally.

[0041] Curcuma zedoaria: This product is the dried rhizome of Curcuma phaeocaulis Val., Curcuma kwangsiensis SGLee et CFLang, or Curcuma wenyujin YHChen et C.Ling, all members of the Zingiberaceae family. It is harvested in winter after the stems and leaves wither, cleaned, steamed or boiled until thoroughly cooked, then sun-dried or low-temperature dried, and the fibrous roots and impurities removed. It has the properties of promoting qi and dissolving blood stasis, dissipating stagnation, and relieving pain. It is used for amenorrhea caused by blood stasis, heart pain due to chest pain, and bloating and pain caused by food stagnation. Take 6-9g orally.

[0042] Clematis: The dried roots and rhizomes of Clematis chinensis Osbeck (Clematishexapetala Pall.) or Clematis manshurica Ruper (Clematishexapetala Pall.) from the Ranunculaceae family are harvested in autumn, the stems and leaves are removed, the soil is washed off, and the skin is sun-dried, or cut into sections and sun-dried. It has the properties of dispelling rheumatism and dredges the meridians. It is used for rheumatic pain, numbness of the limbs, muscle spasms, and difficulty flexing and extending. Take 6-10g orally.

[0043] Example 1:

[0044] (Preparation of granules based on preparation of 1000)

[0045] Table 1

[0046]

[0047] a. According to the ratio in Table 1, salvia miltiorrhiza, rhodiola rosea, dodder seed, asparagus cochinchinensis, fritillaria cirrhosa, zedoaria zedoaria, and clematis root were crushed into 10 mesh, mixed thoroughly, and extracted with 8 times the amount of water at 70°C for 3 times, each extraction time being 1 hour. The extracts were combined and filtered to obtain an aqueous extract;

[0048] b. The aqueous extract in step a was concentrated under reduced pressure at 60°C to an extract with a relative density of 1.10 to obtain an aqueous extract, which was vacuum-dried at 40°C and crushed to obtain an aqueous extract powder;

[0049] c. The extract or extract powder in step b is mixed with 95% ethanol as a binder and auxiliary materials such as sucrose or lactose or starch and dextrin, and the mixture is prepared into granules according to conventional pharmaceutical methods.

[0050] Example 2 (preparation of granules based on preparation of 1000)

[0051] a. According to the ratio in Table 1, respectively crush the salvia miltiorrhiza, rhodiola rosea, dodder seed, asparagus cochinchinensis, fritillaria cirrhosa, zedoaria zedoaria, and clematis root into 100 mesh, mix thoroughly, and extract with 20 times the concentration of 30% ethanol at 80°C for 2 times, each extraction time being 1.5 hours. Combine the extracts and filter to obtain an alcohol extract;

[0052] b. Concentrating the alcohol extract in step a under reduced pressure at 80° C. to an extract with a relative density of 1.10 to obtain an alcohol extract or vacuum drying the alcohol extract at 60° C. and crushing the extract to obtain an alcohol extract powder;

[0053] c. The extract or extract powder in step c is mixed with ethanol as a binder and auxiliary materials such as sucrose or lactose or starch and dextrin, and the mixture is prepared into granules according to conventional pharmaceutical methods.

[0054] Example 3

[0055] (Prepare oral solution based on 1000)

[0056] Table 2

[0057]

[0058] a. According to the ratio in Table 2, salvia miltiorrhiza, rhodiola rosea, dodder seed, asparagus cochinchinensis, fritillaria cirrhosa, zedoaria zedoaria, and clematis root were crushed into 100 mesh, mixed thoroughly, and extracted with 10 times the concentration of 10% ethanol at 60°C for 3 times, each extraction time being 1 hour. The extracts were combined and filtered to obtain an alcohol extract;

[0059] b. Concentrating the alcohol extract in step a under reduced pressure at 40° C. to an extract with a relative density of 1.10 to obtain an alcohol extract or vacuum drying the alcohol extract at 80° C. and crushing the extract to obtain an alcohol extract powder;

[0060] c. Add purified water, 10% stevioside, and 10% sorbic acid to the extract or extract powder prepared in step b, filter through a plate and frame, package, and sterilize to prepare an oral solution.

[0061] Example 4

[0062] (Prepare oral solution based on 1000)

[0063] a. According to the ratio in Table 2, salvia miltiorrhiza, rhodiola rosea, dodder seed, asparagus cochinchinensis, fritillaria cirrhosa, zedoaria zedoaria, and clematis root were crushed into 100 mesh, mixed thoroughly, and extracted with 5 times the amount of water at 80° C. for 3 times, each extraction time being 1.5 hours. The extracts were combined and filtered to obtain an aqueous extract;

[0064] b. Concentrate the water extract in step b under reduced pressure at 60° C. to an extract with a relative density of 1.10 to obtain a water extract or vacuum dry the water extract at 70° C. and grind it to obtain a water extract powder;

[0065] c. Add purified water, stevioside, and sorbic acid to the extract or extract powder in step b, filter through a plate and frame, package, and sterilize to prepare an oral solution.

[0066] Example 5

[0067] (Based on the preparation of 1000, tablets are prepared)

[0068] Table 3

[0069]

[0070] a. According to the ratio in Table 3, salvia miltiorrhiza, rhodiola rosea, dodder seed, asparagus cochinchinensis, fritillaria cirrhosa, zedoaria zedoaria, and clematis root were crushed into 50 mesh, extracted with 10 times the amount of 50% ethanol at 60°C, extracted 4 times, each time for 6 hours, combined the extracts, and filtered to obtain an alcohol extract;

[0071] b. Concentrating the obtained alcohol extract under reduced pressure at 80° C. to an extract with a relative density of 1.05 to obtain an alcohol extract; or vacuum drying the alcohol extract at 60° C. and crushing the extract to obtain an alcohol extract powder;

[0072] c. The extract or extract powder in step b is mixed with 5% cross-linked polyvinyl pyrrolidone 95% ethanol solution as a binder and excipients, and prepared into tablets according to conventional pharmaceutical methods.

[0073] Example 6

[0074] (Prepare tablets based on 1000)

[0075] a. According to the ratio in Table 3, salvia miltiorrhiza, rhodiola rosea, dodder seed, asparagus cochinchinensis, fritillaria cirrhosa, zedoaria zedoaria, and clematis root were crushed into 100 mesh respectively, extracted with 20 times of water, the temperature was 70 ° C, extracted 3 times, each extraction was 1 hour, the extracts were combined, and filtered to obtain an aqueous extract;

[0076] b. Concentrating the obtained aqueous extract at a temperature of 70° C. under reduced pressure to an extract having a relative density of 1.10 to obtain an aqueous extract or vacuum drying the aqueous extract at a temperature of 70° C. and crushing the aqueous extract to obtain an aqueous extract powder;

[0077] c. The extract or extract powder in step c is mixed with 5% cross-linked polyvinyl pyrrolidone ethanol solution as a binder and excipients, and prepared into tablets according to conventional pharmaceutical methods.

[0078] Example 7

[0079] (Based on the preparation of 1000, prepare capsules)

[0080] Table 4

[0081]

[0082]

[0083] a. According to the ratio in Table 4, respectively crush the salvia miltiorrhiza, rhodiola rosea, dodder seed, asparagus cochinchinensis, fritillaria cirrhosa, zedoaria zedoaria, and clematis root into 100 mesh, mix thoroughly, and extract with 10 times the amount of 95% ethanol at 80°C for 4 hours. Filter to obtain the alcohol extract;

[0084] b. Concentrating the obtained alcohol extract under reduced pressure at 60° C. to an extract with a relative density of 1.15 to obtain an alcohol extract or vacuum drying the alcohol extract at 60° C. and crushing the extract to obtain an alcohol extract powder;

[0085] c. The extract or extract powder in step c is mixed with 5% cross-linked polyvinyl pyrrolidone 95% ethanol solution as a binder and excipients, and prepared into capsules according to conventional pharmaceutical methods.

[0086] Example 8

[0087] (Based on the preparation of 1000, prepare capsules)

[0088] a. According to the ratio in Table 4, salvia miltiorrhiza, rhodiola rosea, dodder seed, asparagus cochinchinensis, fritillaria cirrhosa, zedoaria zedoaria, and clematis root were crushed into 100 mesh respectively, mixed thoroughly, and extracted with 25 times the amount of water at 100° C. for 2 times, each time for 2 hours, to obtain an aqueous extract;

[0089] b. Concentrating the obtained aqueous extract under reduced pressure at 60° C. to an extract with a relative density of 1.15 to obtain an aqueous extract; or vacuum drying the aqueous extract at 60° C. and crushing the aqueous extract to obtain aqueous extract powder;

[0090] c. The extract or extract powder in step c is mixed with 5% cross-linked polyvinyl pyrrolidone 95% ethanol solution as an adhesive and excipients, and prepared into capsules according to conventional pharmaceutical methods.

[0091] Embodiment 9

[0092] (Prepare pills based on 1000)

[0093] Table 5

[0094]

[0095] a. According to the ratio in Table 5, salvia miltiorrhiza, rhodiola rosea, dodder seed, asparagus cochinchinensis, fritillaria cirrhosa, zedoaria zedoaria, and clematis root were crushed into 100 mesh respectively, mixed thoroughly, and extracted with 12 times the amount of water at a temperature of 95-100° C. for 3 times, each extraction for 3 hours, and the extracts were combined and filtered to obtain an aqueous extract;

[0096] b. Concentrating the obtained aqueous extract under reduced pressure at 80° C. to an extract with a relative density of 1.40 to obtain an aqueous extract; or vacuum drying the aqueous extract at 80° C. and crushing the aqueous extract to obtain aqueous extract powder;

[0097] c. The extract or extract powder in step c is thoroughly mixed with honey or purified water, and the mixture is made into pills according to conventional pharmaceutical methods. Macrogol 4000 is thoroughly mixed, and the cooling medium is used to make drop pills according to conventional pharmaceutical methods.

[0098] Example 10 (preparation of pills based on 1000)

[0099] a. According to the ratio in Table 5, salvia miltiorrhiza, rhodiola rosea, dodder seed, asparagus cochinchinensis, fritillaria cirrhosa, zedoaria zedoaria, and clematis root were crushed into 80 mesh, mixed thoroughly, and extracted with 20 times the amount of 75% ethanol at 90°C for 3 times, each time for 5 hours. The extracts were combined and filtered to obtain an alcohol extract;

[0100] b. Concentrate the obtained alcohol extract at a temperature of 70° C. to an extract with a relative density of 1.30 to obtain an alcohol extract; or vacuum dry the alcohol extract at a temperature of 70° C. and grind it to obtain a water extract powder;

[0101] c. Fully mix the extract or extract powder in step c with honey or purified water, and make them into pills according to conventional pharmaceutical methods; fully mix polyethylene glycol 4000, and cool the medium with conventional pharmaceutical methods to make them into drop pills.

[0102] Example 11 (Preparation of syrup based on preparation 1000)

[0103] Table 6

[0104]

[0105] a. According to the ratio in Table 6, salvia miltiorrhiza, rhodiola rosea, dodder seed, asparagus cochinchinensis, fritillaria cirrhosa, zedoaria zedoaria, and clematis root were crushed into 100 mesh respectively, and extracted with 20 times the amount of 40% ethanol at 65°C for 3 times, each time for 6 hours. The extracts were combined and filtered to obtain an alcohol extract;

[0106] b. Concentrate the obtained alcohol extract at a temperature of 60° C. to an extract with a relative density of 1.15 to obtain an alcohol extract; or vacuum dry the alcohol extract at a temperature of 60° C. and grind it to obtain an alcohol extract powder;

[0107] c. Add purified water and sucrose to the extract or extract powder in step c, divide and sterilize, and prepare a syrup according to a conventional method.

[0108] Example 12

[0109] (Prepare syrup based on 1000)

[0110] a. According to the ratio in the table of Example 11, Danshen, Rhodiola, Cuscuta, Asparagus, Fritillaria, Curcuma, and Clematidis were crushed into 80 mesh respectively, and extracted with 18 times the amount of water at a temperature of 65 ° C., extracted 3 times for 4 hours each time, combined the extracts, and filtered to obtain an aqueous extract;

[0111] b. Concentrate the obtained water extract at a temperature of 40°C to an extract with a relative density of 1.15 to obtain a water extract; or vacuum dry the water extract at a temperature of 40°C and grind it to obtain a water extract powder;

[0112] c. Add purified water and sucrose to the extract or extract powder in step c, divide the mixture into subpackages, sterilize, and prepare a syrup according to conventional methods.

[0113] Example 13

[0114] (Prepare honey paste based on the preparation of 1000)

[0115] Table 7

[0116]

[0117] a. According to the ratio in Table 7, salvia miltiorrhiza, rhodiola rosea, dodder seed, asparagus cochinchinensis, fritillaria cirrhosa, zedoaria zedoaria, and clematis root were crushed into 65 mesh respectively, and extracted with 12 times the amount of 90% ethanol at 90°C for 2 times, each time for 5 hours. The extracts were combined and filtered to obtain an aqueous extract;

[0118] b. Concentrate the obtained alcohol extract under reduced pressure at 45° C. to obtain an extract with a relative density of 1.40 to obtain an alcohol extract; or vacuum dry the alcohol extract at 45° C. and grind it to obtain an alcohol extract powder;

[0119] c. Add refined honey to the extract or extract powder in step c, stir and mix, divide and package, sterilize, and prepare honey paste according to conventional methods.

[0120] Example 14

[0121] (Prepare honey paste based on the preparation of 1000)

[0122] a. According to the Salvia miltiorrhiza, Rhodiola rosea, Cuscuta chinensis, Asparagus cochinchinensis, Fritillaria cirrhosa, Curcuma zedoaria, and Clematidis in the table of Example 7, they were crushed to 20 mesh, mixed thoroughly, and extracted with 22 times the amount of water at 80°C for 4 times, each for 5 hours. The extracts were combined and filtered to obtain an aqueous extract;

[0123] b. Concentrating the obtained aqueous extract under reduced pressure at a temperature of 55° C. to obtain an extract with a relative density of 1.30 to obtain an aqueous extract or vacuum drying the aqueous extract at a temperature of 55° C. and crushing the aqueous extract to obtain an aqueous extract powder;

[0124] c. Add refined honey to the extract or extract powder of step c, stir and mix, package, sterilize, and prepare a honey paste according to a conventional method.

[0125] Example 15

[0126] The therapeutic effect of the compound extract of the present invention on adjuvant-induced arthritis model in mice:

[0127] Purpose of the experiment:

[0128] This study used modern medical research theories to establish an adjuvant-induced arthritis model in mice and evaluated the therapeutic effect of the compound extract on arthritis mice.

[0129] Experimental Materials

[0130] 2.1 Reagents and Materials

[0131] Table 8 Reagents

[0132]

[0133] 2.2 Instruments and equipment

[0134] Table 9 Instruments

[0135]

[0136]

[0137] 2.3 Experimental Animals: Clean-grade BALB / C57 mice were purchased from the Experimental Animal Center of Xinjiang Medical University. All animal husbandry and experimental procedures were approved by the Animal Welfare and Ethics Committee of Xinjiang University.

[0138] 3. Experimental methods:

[0139] 3.1 Modeling method and grouping

[0140] Establishment and treatment of adjuvant-induced arthritis model in mice: 48 BALB / c male mice were divided into 6 groups, namely untreated group, model group, low-dose DHT treatment group (20 mg / kg), medium-dose DHT group (39 mg / kg), high-dose DHT group (78 mg / kg), and positive control group (methotrexate, MTX, 1 mg / kg), with 8 mice in each group. 25ul of emulsified FCA was injected into the right paw to establish the AIA mouse model. The successful establishment of the model was determined by significant edema in the paw, and recorded as day 0. Treatment began on the first day, with the drug injected subcutaneously into the right posterior buttock. The mice were treated once every 2 days and their weight was weighed. The thickness of the paw, ankle thickness and width were measured. The experiment was terminated after the degree of paw edema changed significantly. The mice were weighed and photographed.

[0141] 3.2 Observation indicators:

[0142] ① The eyeballs were removed and blood was collected. The blood was placed at 37℃ for 1 hour and then centrifuged at 3000 rpm for 20 minutes. The supernatant was collected and the levels of cytokines IL-17A, TNF-α, IL-10, and IFN-γ in the serum were detected by ELISA.

[0143] ② The right leg lymph node of the mouse was taken and ground to prepare a single-cell suspension. The suspension was passed through a mesh into a 1.5 mL centrifuge tube and centrifuged at 1200 rpm for 7 min. The suspension was washed once with PBS and centrifuged again. The supernatant was discarded. After counting the cells, the suspension was stained with flow cytometry antibodies FITC-anti-CD11c, APC-anti-CD40, EV460-anti-F4 / 80, and PE-anti-CD86 in the dark for 15 min. The suspension was washed once with PBS and the status of DCs and macrophages in the lymph nodes was detected by flow cytometry.

[0144] ③ The organs of mice were removed and weighed, and the organ weight / body weight (mg / g) was used as the organ index.

[0145] ④H&E staining: The ankle joint was decalcified, fixed with paraformaldehyde, and then stained with H&E.

[0146] 4. Experimental Results

[0147] Compound extract (DHT) alleviates inflammatory symptoms in AIA mice

[0148] To determine the therapeutic effect of DHT on arthritis, a Freund's adjuvant-induced arthritis (AIA) mouse model was established using different doses of DHT. A low-dose DHT group (20 mg / kg), a medium-dose DHT group (39 mg / kg), and a high-dose DHT group (78 mg / kg) were set up. Methotrexate (MTX) (1 mg / kg) served as a positive control. Edema of the paws and ankles of the mice, pathological changes in the ankles, and serum factor levels were measured. The modeling and treatment process are shown in the table. Figure 4 The results showed that compared with the model group, low-dose and high-dose could significantly reduce the thickness of the sole of the foot (p < 0.001), ankle width (p < 0.001) and ankle thickness (p < 0.01), and alleviate the degree of edema in the sole of the foot and joints ( Figure 1 shown).

[0149] Joint pathological tissue sections showed that in the model group, there was a large number of inflammatory cell infiltration, chondrocyte and osteocyte proliferation, large cell nucleus, and narrowed joint space; in the low-dose DHT treatment group, the state of chondrocyte and osteocyte was significantly improved, the proliferation was reduced, the number of cell nuclei was reduced, and some joint spaces were widened; in the high-dose DHT treatment group, the joint space was widened, the number of inflammatory cells was reduced, the chondrocyte and osteocyte nucleus became regular, and there was no obvious proliferation; in the MTX treatment group, the cells were slightly proliferative, the cell nucleus was slightly larger and darkly stained, some cells had atypia, and the joint space was widened ( Figure 4 ).

[0150] The level of anti-inflammatory cytokine IL-17A in serum was reduced (p>0.05), and the level of IFN-γ was significantly reduced (p<0.001), while the levels of TNF-α and IL-10 had no significant difference (p>0.05). Figure 5 )

[0151] After treatment with DHT, compared with the model group, the medium-dose and high-dose treatment groups significantly reduced the proportion of DC expressing CD40 (CD11c+CD40+) cells in the leg lymph nodes of mice (p < 0.01), and the high-dose treatment group significantly reduced the proportion of DC expressing CD86 (CD11c+CD86+) cells in the leg lymph nodes of mice (p < 0.01), and at the same time reduced the proportion of macrophages dually expressing F4 / 80 and CD40 (CD11b+F4 / 80+CD40+) (p < 0.01) and dually expressing F4 / 80 (CD11b+F4 / 80+CD86+) cells (p < 0.05), indicating that DHT can significantly inhibit the activation state of DC and macrophages ( Figure 6 ).

[0152] Conclusion: The experimental results show that the DHT treatment group can alleviate the inflammatory response and improve joint damage, and the therapeutic effect is better than the positive drug MTX group. This experiment shows that the compound extract has a good anti-rheumatoid arthritis effect.

[0153] Example 16

[0154] The therapeutic effect of the compound extract of the present invention on adjuvant-induced arthritis model in mice:

[0155] Purpose of the experiment:

[0156] This study used modern medical research theories to establish an adjuvant-induced arthritis model in mice and evaluated the therapeutic effect of the compound extract on arthritis mice.

[0157] 2. Experimental Materials, Reagents, and Instruments

[0158] See Table 8 and Table 9 in Example 15.

[0159] 3. Experimental methods and test indicators

[0160] The spleen was ground into a single-cell suspension, passed through a mesh and transferred to a 7 mL centrifuge tube. After centrifugation at 1200 rpm for 7 min, the cells were resuspended in 5 mL of culture medium, counted, and grouped for staining: ① Flow cytometry staining with FITC-anti-CD4, PC5.5-anti-Foxp3, and APC-anti-CD25 antibodies was used to detect the expression of Th1, Th2, Th17, and Treg cell transcription factors. Spleen cells were seeded at 1x106 cells / mL in 24-well plates, with two groups plated for each sample. GolgiStop 0.66 uL / well was added 1 hour later, and the cells were collected 12 hours later. ② Flow cytometry staining with PC7-anti-CD11a, FITC-anti-CD11b, PE-anti-IL-6, and APC-anti-TNF-α antibodies was used to detect DC and macrophage activation and cytokine expression in the spleen. ③ Flow cytometry staining with FE450-anti-CD3, FITC-anti-CD11b, PC5.5-anti-IL-4, PE-anti-IL-17A, and APC-anti-IFN-γ antibodies was used to detect CD4+ T cell-specific cytokine expression levels in the spleen. CytoFLEX flow cytometer was used for detection, and flow cytometric data were analyzed with CytoExpert (24.0.28, Beckman Coulter) software.

[0161] 4. Experimental Results

[0162] 4.1 Compound extract reduces DC and macrophage activation and cytokine expression in the spleen of AIA mice

[0163] After treatment with low-dose, medium-dose and high-dose TDH, the number of DCs that highly expressed IL-6 (CD11 c+IL-6+) (p<0.01) and TNF-a (CD11 c+TNF-a+) (p<0.01) in the spleen was significantly reduced compared with the model group. At the same time, the number of macrophages that highly expressed IL-6 (CD11 b+F4 / 80+IL-6+) (p<0.01) and TNF-a (CD11 b+F4 / 80+TNF-a+) (p<0.01) was also reduced, indicating that TDH can inhibit the activation of DCs and macrophages and downregulate the secretion of proinflammatory cytokines IL-6 and TNF-a ( Figure 7 ).

[0164] 4.2 Compound extract regulates the number of CD4+ cells in the spleen of AIA mice

[0165] The low dose significantly increased the number of Treg (CD40+CD25+Foxp3+) in the spleen of mice (p < 0.05). Figure 8 ).

[0166] 4.3 The compound extract regulates the cytokine expression of CD4+T cells in the spleen of AIA mice. Low-dose, medium-dose and high-dose treatment significantly reduced the number of pro-inflammatory Th1 (CD4+IFN-γ+) (p < 0.001) and Th1 (CD4+IL-17α+) (p < 0.001) cells in the spleen of mice. The MTX treatment group increased the Th1 (CD4+IL-4+) (p < 0.001) that inhibits inflammation. Compared with the model group, there was no significant difference in the TDH treatment group (p > 0.05), indicating that DHT can regulate CD4+T cell differentiation, reduce the number of Th1 and Th17 cells that promote inflammatory response in the spleen of mice, and thus alleviate arthritis ( Figure 9 )

[0167] Conclusion: The experimental results show that the DHT treatment group significantly reduced the number of Th1 and Th17 cells, increased the number of Treg cells, improved the Th2 / Th1 cell ratio, restored the CD4 cell immune balance, and played a therapeutic role.

[0168] It will be appreciated that in the development of any actual embodiment, as in any engineering or design project, numerous implementation-specific decisions may be made. Such a development effort may be complex and time-consuming, but will, for those of ordinary skill having the benefit of this disclosure, be a routine undertaking of design, fabrication, and production without undue experimentation.

[0169] It should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention and are not intended to limit the present invention. Although the present invention has been described in detail with reference to the preferred embodiments, those skilled in the art should understand that the technical solutions of the present invention may be modified or replaced by equivalents without departing from the spirit and scope of the technical solutions of the present invention, which should all be included in the scope of the claims of the present invention.

Claims

1. A compound preparation for treating rheumatoid arthritis, characterized in that: The invention comprises the following raw materials in parts by weight: 5-100 parts of salvia miltiorrhiza, 5-200 parts of rhodiola rosea, 5-100 parts of dodder seeds, 5-100 parts of asparagus cochinchinensis, 5-100 parts of fritillaria cirrhosa, 5-100 parts of zedoaria, 5-100 parts of clematis root, and medically acceptable excipients including 1-50 parts of lactose, 1-5 parts of starch, 5-100 parts of sucrose, 1-50 parts of dextrin, 1-50 parts of pregelatinized starch, 5-100 parts of microcrystalline cellulose, 5-100 parts of cross-linked polyvinyl pyrrolidone, 5-100 parts of low-substituted hydroxypropyl cellulose, 5-100 parts of sodium carboxymethyl starch, 1-50 parts of talc, 1-50 parts of micropowdered silica gel, 1-50 parts of magnesium stearate, 5-100 parts of polyethylene glycol, 10-200 parts of honey, 1-50 parts of stevioside, and 10-50 parts of sorbic acid. Made into granules, oral preparations, pills, tablets, capsules, syrups, and honey pastes.

2. A compound preparation for treating rheumatoid arthritis according to claim 1, characterized in that: The oral preparations prepared from the compound preparation include but are not limited to pills, tablets, capsules, syrups, and honey pastes.

3. The method for preparing a compound preparation for treating rheumatoid arthritis according to claim 1, wherein: The following steps are involved: S1. Grind 5-100 parts of Salvia miltiorrhiza, 5-200 parts of Rhodiola rosea, 5-100 parts of Cuscuta chinensis, 5-100 parts of Asparagus cochinchinensis, 5-100 parts of Fritillaria cirrhosa, 5-100 parts of Curcuma zedoaria, and 5-100 parts of Clematidis to 10-100 mesh size, mix, and extract with 5-25 times the amount of water or 10%-95% ethanol at a temperature of 10-100° C. for 1-4 times, each extraction time being 1-6 hours. Combine the extracts and filter to obtain a water extract or an alcohol extract. S2. The water extract or alcohol extract in step a is concentrated under reduced pressure at a temperature of 40-80° C. to an extract with a relative density of 1.10-1.40 to obtain a water extract or an alcohol extract, and the water extract or the alcohol extract is vacuum dried at a temperature of 10-80° C. and crushed to obtain a water extract powder or an alcohol extract powder; Alternatively, the water-extracted extract powder or alcohol-extracted extract powder in step b is mixed with 10%-95% ethanol as a binder and auxiliary materials such as sucrose, lactose or starch 10-200 parts and dextrin 10-100 parts, and the mixture is thoroughly mixed and prepared into granules according to conventional pharmaceutical methods. Alternatively, the water-extracted extract powder or the alcohol-extracted extract powder in step b is added with purified water, 1-50 parts of stevia, and 1-50 parts of sorbic acid, and the mixture is filtered on a plate and frame, packaged, and sterilized to prepare an oral solution; Alternatively, the water-extracted extract powder or the alcohol-extracted extract powder in step b is fully mixed with 10-200 parts of honey and 5-100 parts of polyethylene glycol as auxiliary materials, and the mixture is prepared into pills according to conventional pharmaceutical methods. Alternatively, the water-extracted extract powder or alcohol-extracted extract powder in step b is mixed with 1%-10% cross-linked polyvinyl pyrrolidone and 10%-95% ethanol solution as a binder, and excipients including 1-50 parts of lactose, 5-100 parts of microcrystalline cellulose, 5-100 parts of low-substituted hydroxypropyl cellulose, 5-100 parts of sodium carboxymethyl starch, 1-50 parts of pregelatinized starch, 1-50 parts of micropowdered silica gel, 1-50 parts of magnesium stearate, and 1-50 parts of talc, and prepared into tablets or capsules according to conventional pharmaceutical methods; Alternatively, the water-extracted extract powder or the alcohol-extracted extract powder in step b is added with purified water, mixed thoroughly with 5-100 parts of sucrose as an auxiliary material, filtered, packaged, sterilized, and prepared into a syrup according to conventional methods; Alternatively, the water-extracted extract powder or the alcohol-extracted extract powder in step b is added with 10-200 parts of honey, stirred and mixed, packaged, sterilized, and prepared into a honey paste according to a conventional method.

4. The compound preparation for treating rheumatoid arthritis according to claim 1 is used to treat rheumatoid arthritis, ankylosing spondylitis, chronic arthritis, and rheumatic sciatica.

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