Combined medicine for treating rheumatoid arthritis and application and medicine box thereof

Abstract

The invention provides a combined medicine for treating rheumatoid arthritis as well as application and a medicine box thereof, and belongs to the field of medicines. The combined medicine disclosed by the invention comprises (a) a traditional Chinese medicine composition with an effective treatment amount, wherein the traditional Chinese medicine composition comprises the following raw material medicines: tortoise-plastron glue, deer-horn glue, morinda officinalis, fructus psoraleae, teasel root, eucommia ulmoides, prepared rehmannia root, angelica sinensis, radix paeoniae alba, fructus lycii, schisandra chinensis, Chinese yam, dogwood, radix ophiopogonis, gordon euryale seed, cinnamon, radix aconiti carmichaeli, moutan bark, rhizoma alismatis, poria cocos and an optional pharmaceutically acceptable carrier or excipient; (b) a therapeutically effective amount of methotrexate or a pharmaceutically acceptable derivative thereof, and optionally, a pharmaceutically acceptable carrier or excipient. The traditional Chinese medicine composition is combined with methotrexate for use, so that the treatment effect of rheumatoid arthritis can be improved, the dosage of methotrexate can be reduced, adverse reactions caused by long-term administration of methotrexate can be reduced, and the compliance of a patient can be improved under the condition that the patient takes the methotrexate for a long time.

Description

Technical Field

[0001] The present disclosure relates to the field of medicines, and in particular to a combined medicine for treating rheumatoid arthritis and an application thereof, and a medicine kit. Background Art

[0002] Rheumatoid arthritis (RA) is a chronic, inflammatory autoimmune disease with a complex etiology, which is mainly manifested by inflammation of the synovium, damage to articular cartilage and bone, often involving large and small joints of the hands and feet, and can cause joint deformity and dysfunction in severe cases. There is currently no effective cure for RA, and clinically, the treatment methods are mainly to suppress immune and inflammatory symptoms and relieve pain symptoms. In the pathological process of RA, inflammatory response, autoimmune imbalance, and abnormal proliferation of synovial cells are important factors in joint bone tissue damage. At present, clinical treatment of Western medicine mainly uses anti-rheumatic drugs, non-steroidal anti-inflammatory drugs, glucocorticoids and biological agents to treat this disease.

[0003] There are many types of disease modifying anti-rheumatic drugs (DMARDs), and their mechanisms of action are different, including methotrexate, tumor necrosis factor-α (TNF-α) inhibitors, tofacitinib, etc. When used properly, they can delay or prevent joint damage and bone loss caused by RA, and can effectively improve the living standards of RA patients. However, DMARDs cannot directly treat joint pain and swelling, and may induce more serious adverse reactions, including gastrointestinal damage, liver toxicity, kidney damage, stomatitis, bone marrow suppression, etc. For example: Over the years, many studies around the world have repeatedly reported the limitations of methotrexate in the treatment of RA, including gastrointestinal reactions, abnormal elevation of transaminases, hair loss and other adverse reactions after long-term use, and more than 40% of patients treated with methotrexate showed poor efficacy after two years. In addition, although there are biosynthetic DMARDs and targeted DMARDs with fewer adverse reactions, these drugs are usually expensive and are only used as alternative drugs after traditional DMARDs are ineffective. Nonsteroidal anti-inflammatory drugs (NSAIDs) are usually used in the early stage of RA because they can alleviate the clinical symptoms of RA patients, eliminate local inflammatory reactions in joints, and effectively relieve joint pain, but they cannot prevent the development of joint damage. DMARDs must be added as soon as possible to effectively control the disease. In addition, NSAIDs may induce adverse reactions such as peptic ulcers and bleeding. Long-term oral administration can cause serious damage to the gastrointestinal tract, and they are rarely used alone in clinical practice. Glucocorticoids have a rapid onset of action in the treatment of RA. They can be used to control acute lesions, delay early joint destruction in RA, and improve joint function. They are often used in combination with DMARDs. However, glucocorticoids cannot cure RA. RA is prone to relapse after discontinuation of the drug. In addition, there are more difficult-to-treat adverse drug reactions when used for a long time, including osteoporosis, hypertension, hormonal obesity, fluid retention, hyperglycemia, cataracts, decreased skin elasticity, and atherosclerosis.

[0004] Therefore, there is an urgent need for a drug for treating rheumatoid arthritis that can effectively inhibit inflammation, inhibit synovial tissue proliferation, and inhibit excessive immune response. Summary of the invention

[0005] The purpose of the present invention is to overcome the deficiencies of the prior art and provide a combined drug for treating rheumatoid arthritis and its application and medicine kit.

[0006] To achieve the above objectives, the technical solution adopted by the present disclosure is:

[0007] In a first aspect, a combination drug for treating rheumatoid arthritis is provided, comprising:

[0008] (a) a Chinese medicine composition containing the following raw materials in a therapeutically effective amount: tortoise shell glue, antler glue, Morinda officinalis, psoralea corylifolia, Dipsacus asper, Eucommia ulmoides, Rehmannia root, Angelica sinensis, White peony root, Lycium barbarum, Schisandra chinensis, Chinese yam, Cornus officinalis, Ophiopogon japonicus, Euryale ferox, Cinnamon bark, Aconite root, Paeonia suffruticosa moutan, Alisma orientalis, Poria, and, optionally, a pharmaceutically acceptable carrier or excipient;

[0009] (b) a therapeutically effective amount of methotrexate or a pharmaceutically acceptable derivative thereof, and, optionally, a pharmaceutically acceptable carrier or excipient.

[0010] In one embodiment, the traditional Chinese medicine composition includes the following raw materials in parts by weight: 1-5 parts of tortoise shell glue, 1-5 parts of deer antler glue, 5-10 parts of Morinda officinalis, 2-8 parts of Psoralea corylifolia, 5-10 parts of Dipsacus asper, 5-10 parts of Eucommia ulmoides, 10-15 parts of Rehmannia root, 5-10 parts of Angelica sinensis, 5-10 parts of White Peony Root, 2-8 parts of Lycium barbarum, 1-5 parts of Schisandra chinensis, 10-20 parts of Dioscorea opposita, 5-15 parts of Cornus officinalis, 2-8 parts of Ophiopogon japonicus, 10-15 parts of Euryale ferox, 1-5 parts of cinnamon, 2-8 parts of Aconite, 5-15 parts of Paeonia suffruticosa, 5-15 parts of Alisma orientalis, and 5-15 parts of Poria.

[0011] In one embodiment, the tortoise shell glue and deer antler glue need to be melted.

[0012] In a second aspect, a use of the combined drug in the preparation of a drug for treating rheumatoid arthritis is provided.

[0013] In a third aspect, a medicine kit for treating rheumatoid arthritis is provided, the medicine kit comprising at least one first preparation unit and at least one second preparation unit, the first preparation unit and the second preparation unit both being independently present in the medicine kit;

[0014] The first preparation unit comprises a therapeutically effective amount of a Chinese medicine composition containing the following raw materials: tortoise shell glue, deer horn glue, Morinda officinalis, psoralea corylifolia, Dipsacus asper, Eucommia ulmoides, Rehmannia root, Angelica sinensis, White peony root, Lycium barbarum, Schisandra chinensis, Chinese yam, Cornus officinalis, Ophiopogon japonicus, Euryale ferox, Cinnamon bark, Aconite root, Paeonia suffruticosa moutan, Alisma orientalis, Poria cocos, and, optionally, a pharmaceutically acceptable carrier or excipient;

[0015] The second dosage unit comprises a therapeutically effective amount of methotrexate or a pharmaceutically acceptable derivative thereof, and, optionally, a pharmaceutically acceptable carrier or excipient.

[0016] In one embodiment, the traditional Chinese medicine composition includes the following raw materials in parts by weight: 1-5 parts of tortoise shell glue, 1-5 parts of deer antler glue, 5-10 parts of Morinda officinalis, 2-8 parts of Psoralea corylifolia, 5-10 parts of Dipsacus asper, 5-10 parts of Eucommia ulmoides, 10-15 parts of Rehmannia root, 5-10 parts of Angelica sinensis, 5-10 parts of White Peony Root, 2-8 parts of Lycium barbarum, 1-5 parts of Schisandra chinensis, 10-20 parts of Dioscorea opposita, 5-15 parts of Cornus officinalis, 2-8 parts of Ophiopogon japonicus, 10-15 parts of Euryale ferox, 1-5 parts of cinnamon, 2-8 parts of Aconite, 5-15 parts of Paeonia suffruticosa, 5-15 parts of Alisma orientalis, and 5-15 parts of Poria.

[0017] In one embodiment, the dosage form of the Chinese medicine composition includes one of tablets, powders, granules, pills, capsules, solutions, emulsions, and injections.

[0018] In one embodiment, the medicine kit further comprises instructions, which stipulate that the first preparation unit and the second preparation unit are preparations for combined use in the treatment of rheumatoid arthritis.

[0019] Compared with the prior art, the beneficial effects of the present invention are as follows: the inventors of the present invention have found through repeated exploration and a large number of experiments, and through clinical application, that the Chinese medicine composition with tortoise shell glue, deer antler glue, Morinda officinalis, Psoralea corylifolia, Dipsacus asper, Eucommia ulmoides, Rehmannia root, Angelica sinensis, White Peony Root, Lycium barbarum, Schisandra chinensis, Dioscorea batatas, Cornus officinalis, Ophiopogon japonicus, Euryale ferox, Cinnamon bark, Aconite root, Paeonia suffruticosa, Alisma orientalis, and Poria as main medicinal materials, combined with methotrexate or its pharmaceutically acceptable derivatives, has a synergistic effect in the treatment of rheumatoid arthritis. The combination of the two can improve the therapeutic efficacy of rheumatoid arthritis, reduce the dosage of methotrexate or its pharmaceutically acceptable derivatives, and thus reduce the adverse reactions caused by methotrexate or its pharmaceutically acceptable derivatives themselves. DETAILED DESCRIPTION

[0020] With reference to the detailed description of the following embodiments, it will be easier to understand the advantages and features of the present disclosure and the methods for achieving the advantages and features. However, the present disclosure can be implemented in various other forms and should not be construed as being limited to the embodiments set forth herein. The embodiments disclosed herein are provided so that the disclosure of the present disclosure will be thorough and complete, and the scope of the present disclosure will be fully conveyed to those skilled in the art.

[0021] The terms used in the specification are intended to describe certain embodiments only and should by no means limit the present disclosure.Expressions in the singular include the meaning of the plural form unless clearly used otherwise.

[0022] Unless otherwise defined, all terms (including technical and scientific terms) used herein have the same meaning as those commonly understood by ordinary technicians in the field to which the present disclosure belongs. Any term defined in a comprehensive dictionary should be interpreted as having the same meaning in the context of the relevant field, and, unless explicitly defined otherwise, should not be interpreted as having an idealistic or overly formalistic meaning.

[0023] In a first aspect, a combination drug for treating rheumatoid arthritis is provided, comprising:

[0024] (a) a Chinese medicine composition containing the following raw materials in a therapeutically effective amount: tortoise shell glue, antler glue, Morinda officinalis, psoralea corylifolia, Dipsacus asper, Eucommia ulmoides, Rehmannia root, Angelica sinensis, White peony root, Lycium barbarum, Schisandra chinensis, Chinese yam, Cornus officinalis, Ophiopogon japonicus, Euryale ferox, Cinnamon bark, Aconite root, Paeonia suffruticosa moutan, Alisma orientalis, Poria, and, optionally, a pharmaceutically acceptable carrier or excipient;

[0025] (b) a therapeutically effective amount of methotrexate or a pharmaceutically acceptable derivative thereof, and, optionally, a pharmaceutically acceptable carrier or excipient.

[0026] RA is classified as "arthralgia" in TCM. "Suwen·Arthralgia" records: "Wind, cold and dampness are mixed together and become arthralgia". The basic cause of RA is that cold and dampness invade the kidneys deeply, causing the tendons and bones to lose their nourishment, leading to loose bones, tendon contractures, and joint deformation. Over time, the meridians will be blocked, and phlegm and blood stasis will be mixed, which will aggravate the condition. Therefore, TCM treatment focuses on tonifying the kidney to treat "arthralgia", which has sufficient theoretical development and clinical research basis. "The Essence of Medical Classics" says: "The kidneys store essence, essence produces marrow, and marrow produces bones, so bones are what the kidneys combine with", "Marrow is produced by kidney essence, and if the essence is sufficient, the marrow is sufficient, and the marrow is in the bones, and if the marrow is sufficient, the bones are strong." The tortoise shell glue in the disclosed Chinese medicine preparation nourishes yin and suppresses yang, benefits the kidney and strengthens bones; the deer antler glue warms and nourishes the liver and kidney, benefits the essence and nourishes blood; the Morinda officinalis nourishes the kidney yang, strengthens the tendons and bones, and eliminates rheumatism; the Rehmannia root nourishes yin, benefits the kidney, and replenishes blood; the Psoralea corylifolia warms the kidney and assists yang, consolidates the essence and reduces urine; the Dipsacus asper nourishes the liver and kidney, strengthens the tendons and bones; the Eucommia ulmoides nourishes the liver and kidney, strengthens the tendons and bones, consolidates the menstruation and stabilizes the pregnancy; the Chinese angelica nourishes blood and activates blood circulation, regulates menstruation and relieves pain; the White Peony Root nourishes blood and regulates menstruation, astringes yin and stops sweating, softens the liver and relieves pain, and calms the liver yang; the Lycium barbarum nourishes the liver and kidney, benefits the essence and improves eyesight; the Schisandra chinensis astringes and consolidates Astringent, invigorating Qi and promoting fluid production, tonifying the kidney and calming the mind. Chinese yam invigorates the spleen and stomach, promotes fluid production and benefits the lungs, invigorates the kidney and astringes the essence. Cornus officinalis invigorates the liver and kidney, astringes and consolidates the body. Ophiopogon japonicus moistens the lungs and nourishes Yin, invigorates the stomach and promotes fluid production, clears the heart and relieves restlessness. Gorgon fruit invigorates the kidney and consolidates the essence, invigorates the spleen and stops diarrhea, removes dampness and stops leukorrhea. Cinnamon bark warms the middle and dispels cold, regulates Qi and relieves pain. Alisma orientalis promotes diuresis and eliminates dampness, relieves heat, clears turbidity and reduces lipids. Aconitum restores yang and rescues from adverse conditions, invigorates fire and assists yang, dispels cold and relieves pain. Paeonia suffruticosa clears heat and cools blood, promotes blood circulation and removes stasis. Combining these herbs can warm and invigorate the kidney yang, replenish the essence and improve the marrow, thereby strengthening the tendons and bones.

[0027] The disclosed Chinese medicine preparation nourishes the kidney and strengthens the marrow, is beneficial to the treatment of "arthritis" in the middle-aged and elderly people with kidney deficiency, and strengthens the tendons and bones; it can significantly relieve inflammation and pain, and inhibit synovial hyperplasia and osteoporosis, taking into account the multiple effects of analgesia, anti-inflammatory and repair, and is suitable for the treatment of RA under different disease progression; the disclosed Chinese medicine composition takes into account both nourishment and treatment, has few adverse reactions, and is used in combination with traditional DMARDs (such as methotrexate), by downregulating the level of estradiol hormone in RA patients, and upregulating the levels of progesterone and testosterone hormones, thereby promoting the expression and action of immunosuppressive cytokines and anti-inflammatory factors in the body, and at the same time, by stimulating chondrocyte growth and calcium supplementation, promoting cartilage repair, and the multiple aspects work together to achieve the therapeutic effect of treating RA; in addition, the Chinese medicine composition is used in combination with methotrexate, while improving the efficacy of the drug, reducing the dosage of methotrexate, reducing the adverse reactions caused by long-term use of methotrexate, and is beneficial to improving the compliance of patients when they take the drug for a long time.

[0028] In the present disclosure, the Chinese medicine composition can be a Chinese patent medicine, specifically with tortoise shell glue, deer antler glue, Morinda officinalis, Psoralea corylifolia, Dipsacus asper, Eucommia ulmoides, Rehmannia root, Angelica sinensis, White Peony Root, Lycium barbarum, Schisandra chinensis, Dioscorea batatas, Cornus officinalis, Ophiopogon japonicus, Euryale ferox, Cinnamon bark, Aconite root, Paeonia suffruticosa moutan, Alisma orientalis and Poria cocos as main functional medicinal materials, under the guidance of traditional Chinese medicine theory, for the purpose of preventing and treating diseases, it is processed into a Chinese medicine product of a certain dosage form according to the prescribed prescription and preparation process.

[0029] In the present disclosure, "combination drug" means drugs that can be used for treatment by the route of combined use of drugs.

[0030] In the present disclosure, the generic name or trade name of a drug merely indicates its active ingredient or a substance that acts as an effective ingredient and should not be considered as limiting the source of the drug.

[0031] In the present disclosure, "carrier" or "excipient" can be any conventional carrier and excipient in the art, and can be specifically selected according to the administration method or disease type and state for treating the patient.

[0032] Specifically, the "carrier" or "excipient" used in the present disclosure may be at least one of refined honey, starch or starch derivatives, and microcrystalline cellulose.

[0033] In the present disclosure, "effective amount" refers to the amount of the active pharmaceutical ingredient in the present disclosure, which can effectively treat rheumatoid arthritis in the present disclosure.

[0034] In one embodiment, the traditional Chinese medicine composition includes the following raw materials in parts by weight: 1-5 parts of tortoise shell glue, 1-5 parts of deer antler glue, 5-10 parts of Morinda officinalis, 2-8 parts of Psoralea corylifolia, 5-10 parts of Dipsacus asper, 5-10 parts of Eucommia ulmoides, 10-15 parts of Rehmannia root, 5-10 parts of Angelica sinensis, 5-10 parts of White Peony Root, 2-8 parts of Lycium barbarum, 1-5 parts of Schisandra chinensis, 10-20 parts of Dioscorea opposita, 5-15 parts of Cornus officinalis, 2-8 parts of Ophiopogon japonicus, 10-15 parts of Euryale ferox, 1-5 parts of cinnamon, 2-8 parts of Aconite, 5-15 parts of Paeonia suffruticosa, 5-15 parts of Alisma orientalis, and 5-15 parts of Poria.

[0035] In one embodiment, the tortoise shell glue and deer antler glue need to be melted.

[0036] In the present disclosure, "melting" means dissolving the gelatinous drug with a hot solution, which may be boiled water, hot rice wine, hot milk, or a hot liquid medicine from which the residue is filtered out. Melting can prevent the gelatinous drug from adhering to other drugs or medicine bottles to avoid burning.

[0037] Specifically, the melting step of the tortoise shell glue and deer horn glue disclosed in the present invention is: adding 1-5 parts by weight of tortoise shell glue and 1-5 parts by weight of deer horn glue into 1-5 parts by weight of white wine and evaporating.

[0038] Specifically, the alcohol content of the liquor is 50-60 degrees.

[0039] In one embodiment, the Morinda officinalis, Psoralea corylifolia and Eucommia ulmoides need to be fried with salt.

[0040] Specifically, aconite contains aconitine, therefore, the aconite used in the present invention is processed aconite slices to reduce the toxicity of aconite, thereby improving the safety of the traditional Chinese medicine composition.

[0041] In a second aspect, a use of the combined drug in the preparation of a drug for treating rheumatoid arthritis is provided.

[0042] In a third aspect, a medicine kit for treating rheumatoid arthritis is provided, the medicine kit comprising at least one first preparation unit and at least one second preparation unit, the first preparation unit and the second preparation unit both being independently present in the medicine kit;

[0043] The first preparation unit comprises a therapeutically effective amount of a Chinese medicine composition containing the following raw materials: tortoise shell glue, deer horn glue, Morinda officinalis, psoralea corylifolia, Dipsacus asper, Eucommia ulmoides, Rehmannia root, Angelica sinensis, White peony root, Lycium barbarum, Schisandra chinensis, Chinese yam, Cornus officinalis, Ophiopogon japonicus, Euryale ferox, Cinnamon bark, Aconite root, Paeonia suffruticosa moutan, Alisma orientalis, Poria cocos, and, optionally, a pharmaceutically acceptable carrier or excipient;

[0044] The second dosage unit comprises a therapeutically effective amount of methotrexate or a pharmaceutically acceptable derivative thereof, and, optionally, a pharmaceutically acceptable carrier or excipient.

[0045] In one embodiment, the traditional Chinese medicine composition includes the following raw materials in parts by weight: 1-5 parts of tortoise shell glue, 1-5 parts of deer antler glue, 5-10 parts of Morinda officinalis, 2-8 parts of Psoralea corylifolia, 5-10 parts of Dipsacus asper, 5-10 parts of Eucommia ulmoides, 10-15 parts of Rehmannia root, 5-10 parts of Angelica sinensis, 5-10 parts of White Peony Root, 2-8 parts of Lycium barbarum, 1-5 parts of Schisandra chinensis, 10-20 parts of Dioscorea opposita, 5-15 parts of Cornus officinalis, 2-8 parts of Ophiopogon japonicus, 10-15 parts of Euryale ferox, 1-5 parts of cinnamon, 2-8 parts of Aconite, 5-15 parts of Paeonia suffruticosa, 5-15 parts of Alisma orientalis, and 5-15 parts of Poria.

[0046] In one embodiment, the dosage form of the Chinese medicine composition includes one of tablets, powders, granules, pills, capsules, solutions, emulsions, and injections.

[0047] In one embodiment, the Chinese medicine composition and methotrexate or a pharmaceutically acceptable derivative thereof can be administered to a subject in need of treatment simultaneously, substantially simultaneously, sequentially, timed, or sequentially.

[0048] In the present disclosure, "first preparation unit" and "second preparation unit" may refer to pharmaceutical preparations or pharmaceutical dosage forms, which may or may not include pharmaceutical excipients. In addition, the amount of active ingredients therein may be changed according to the actual dosage level and the process of administration, and the amount may effectively obtain the desired therapeutic response for a specific subject, composition and administration method, and is non-toxic to the subject. In other words, the specific dosage of the "first preparation unit" and the "second preparation unit" may be determined by those skilled in the art based on prior knowledge.

[0049] In one embodiment, the medicine kit further comprises instructions, which stipulate that the first preparation unit and the second preparation unit are preparations for combined use in the treatment of rheumatoid arthritis.

[0050] The present disclosure is further described below by means of specific examples. However, it should be understood that these examples are only used for more detailed description and should not be construed as limiting the present disclosure in any form.

[0051] In the following examples, "parts" refer to parts by weight; the Chinese medicine preparation is homemade, and its preparation method is as follows:

[0052] (1) adding 5 parts of tortoise shell glue and 5 parts of deer antler glue to 5 parts of white wine and evaporating the mixture, and then uniformly mixing the mixture with 48 parts of refined honey and 16 parts of water to obtain refined honey water A;

[0053] (2) Grind 8 parts of Morinda officinalis, 5 parts of Psoralea corylifolia, 8 parts of Dipsacus asper, 8 parts of Eucommia ulmoides, 12 parts of Rehmannia root, 7 parts of Angelica sinensis, 7 parts of White Peony Root, 5 parts of Lycium barbarum, 3 parts of Schisandra chinensis, 15 parts of Dioscorea opposita, 10 parts of Cornus officinalis, 6 parts of Ophiopogon japonicus, 12 parts of Euryale ferox, 3 parts of Cinnamon bark, 6 parts of Aconite root, 10 parts of Paeonia suffruticosa, 10 parts of Alismatis orientalis, and 10 parts of Poria cocos, and pass through a 50-mesh sieve to obtain Powder B;

[0054] (3) Mix the refined honey water A and the powder B evenly to obtain a Chinese medicine preparation.

[0055] In the following examples, methotrexate tablets were purchased from Shanghai SPH Pharmaceuticals Co., Ltd., with the national medicine approval number H31020644.

[0056] Example 1 Pharmacological test of combined drugs

[0057] 1.1. Tests to regulate sex hormone levels

[0058] Take 100 SPF SD rats weighing 180g-220g, half male and half female. Male and female rats were randomly divided into a control group, a model group, a Chinese medicine preparation group disclosed in the present invention, a methotrexate group, and a methotrexate and Chinese medicine preparation combination group, with 10 rats in each group. After 7 days of adaptive feeding, 100 rats were modeled except for the control group. The model used in the experiment was adjuvant rheumatoid arthritis. The specific method was as follows: a No. 6 syringe was carefully inserted in the right hind limb toe position of the rat at about 15°, and 100 μL of complete Freund's adjuvant was injected. After 7 days, complete Freund's adjuvant was injected again according to the above method. After 24 hours, the rat ankle was inflammatory and swollen, indicating that the model was successful. The control group rats were injected with an equal amount of normal saline.

[0059] After successful modeling, medication was started. Except for methotrexate, which was administered twice a week, the other groups were gavaged daily for 7 days. The methotrexate group was gavaged at a dose of 0.90 mg / kg, twice a week, and the first dose was doubled; the Chinese medicine preparation group was gavaged at a dose of 15g of raw medicine / kg, and the first dose was doubled; the methotrexate and Chinese medicine preparation group was gavaged at a dose of 0.45 mg / kg on the basis of the Chinese medicine preparation group's dosing regimen, twice a week, and the first dose was doubled; the control group and the model group were given an equal volume of normal saline. One day after the last administration, arterial blood was taken from each group of mice, and serum was taken to measure testosterone, progesterone and estradiol in serum by enzyme-linked immunosorbent assay, and then statistical analysis was performed. The results are shown in Table 1-2.

[0060] Table 1 Results of the test on sex hormone level regulation in SD male mice ( n=10)

[0061]

[0062] Note: *P<0.05, **P<0.01 compared with the control group; △P<0.05, △△P<0.01 compared with the model group.

[0063] Table 2 Results of the sex hormone level regulation test in SD female mice ( n=10)

[0064]

[0065]

[0066] Note: *P<0.05, **P<0.01 compared with the control group; △P<0.05, △△P<0.01 compared with the model group.

[0067] It can be seen from the results in Tables 1 and 2 that the Chinese medicine preparation disclosed in the present invention can significantly regulate the sex hormone levels of AIA rats to return to normal levels or approach normal levels (P<0.01).

[0068] 1.2 Anti-inflammatory test

[0069] Take 180g-220g SPF grade SD rats, 70, half male and half female, and randomly divided into control group, model group, disclosed Chinese medicine preparation group, methotrexate group, methotrexate and Chinese medicine preparation combination group, indomethacin tablet group, dexamethasone tablet group, 10 in each group. After 7d of adaptive feeding, 70 rats, except for the control group, all other rats were modeled. The model used in the test is adjuvant rheumatoid arthritis, and the specific method is: carefully insert the No. 6 syringe in the right hind limb toe position of the rat at about 15 ° direction, inject 100 μL of complete Freund's adjuvant, after 7d, inject complete Freund's adjuvant according to the above method again, after 24h, rat ankle inflammatory swelling, indicating successful modeling. Control group rats were injected with an equal amount of normal saline.

[0070] After successful modeling, medication was started. Except for methotrexate, which was administered twice a week, the other groups were gavaged daily for 7 days. The methotrexate group was gavaged at a dose of 0.90 mg / kg, twice a week, and the first dose was doubled; the Chinese medicine preparation group was gavaged at a dose of 15 g of crude drug / kg, and the first dose was doubled; the methotrexate and Chinese medicine preparation combination group was gavaged at a dose of 0.45 mg / kg on the basis of the Chinese medicine preparation group's dosing regimen, twice a week, and the first dose was doubled; the indomethacin tablet test group was gavaged at a dose of 2.5 mg / kg, and the first dose was doubled; the dexamethasone tablet test group was gavaged at a dose of 0.25 mg / kg, and the first dose was doubled; the control group and the model group were given an equal volume of normal saline. One day after the last administration, the volume of plantar swelling was recorded with a rat plantar volume measurement device, and the swelling inhibition rate was calculated; and arterial blood was taken to measure the TNFα concentration in serum by enzyme-linked immunosorbent assay. The results are shown in Table 3.

[0071] Swelling rate = (average drainage volume - average drainage volume of control group) / average drainage volume of control group × 100%; swelling inhibition rate = (average swelling rate of model group - average swelling rate of drug-treated group) / average swelling rate of model group × 100%.

[0072] Table 3 Anti-inflammatory test results ( n=10)

[0073]

[0074]

[0075] Note: *P<0.05, **P<0.01 compared with the control group; △P<0.05, △△P<0.01 compared with the model group.

[0076] From the results in Table 3, it can be seen that the combination of the Chinese medicine preparation disclosed in the present invention and methotrexate has a significant inhibitory effect on the swelling of the ankle and the secretion of inflammatory factors in rats, and can effectively alleviate the inflammatory symptoms of adjuvant-induced RA rats (P<0.01).

[0077] 1.3 Autoimmune suppression test

[0078] Blood samples of rats in each group in the anti-inflammatory test of 1.2 were taken, placed in empty tubes, incubated at 27°C for 120 min, washed with washing solution, added with horseradish peroxidase, washed again, and finally measured with a combined immunoassay reader to detect serum immunoglobulins Ig A, IgG, and Ig M. The results are shown in Table 4.

[0079] Table 4 Ig A, IgG, Ig M expression levels (ng / ml, n=10)

[0080]

[0081] Note: *P<0.05, **P<0.01 compared with the control group; △P<0.05, △△P<0.01 compared with the model group.

[0082] Take 25 μL of blood sample from each group of rats in the anti-inflammatory test in 1.2, and 6 Add 0.25 μg of anti-mouse CD4-FITC and 1.0 μg of anti-mouse CD25-PE to each cell / tube, mix well, and keep away from light for 20-30 min; add 1 mL of red blood cell lysis solution, and keep away from light for 15-25 min; wash twice with PBA, centrifuge and discard the supernatant, and detect the proportion of CD4+Treg and CD4+CD25+Treg by flow cytometry. The results are shown in Table 5.

[0083] Table 5 Expression of CD4+Treg and CD4+CD25+Treg (%, n=10)

[0084]

[0085] Note: *P<0.05, **P<0.01 compared with the control group; △P<0.05, △△P<0.01 compared with the model group.

[0086] The results in Tables 4 and 5 show that the combination of the Chinese medicine preparation of the present invention and methotrexate can significantly inhibit the levels of serum immunoglobulins Ig A, IgG, and Ig M in RA rats, and promote the increase in the expression ratios of CD4+Treg and CD4+CD25+Treg in rat serum, indicating that the combined drug of the present invention can regulate immune function and inhibit RA-induced autoimmune reactions.

[0087] 1.4 Joint repair test

[0088] Healthy SD rats were taken to extract primary ankle chondrocytes and synovial cells according to the primary cell extraction process and cultured in 6-well plates for later use. The drug-containing serum of rats in each group in the anti-inflammatory test in 1.2 was taken and administered in the culture medium at a dose of 15% drug-containing serum. The cells were incubated at 37°C and 5% CO2 for 48 hours. The viability of chondrocytes and synovial cells was detected by thiazole blue colorimetry, and the effects of each group of drugs on the proliferation of chondrocytes and synovial cells were compared. The results are shown in Table 6.

[0089] Blood samples of rats in each group in the anti-inflammatory test in 1.2 were taken to prepare serum, and the levels of RANKL and OPG in rat serum were detected by ELISA according to the instructions of the kit. 100 μL of serum from each group of rats was added to the wells of the enzyme-labeled plate coated with RANKL or OPG antibody, incubated at 37°C for 1 hour, washed with washing solution, added with biotin 2 antibody conjugate, incubated at 37°C for 1 hour, washed, added with avidin 2 enzyme conjugate, incubated at 37°C for 30 minutes, finally washed, added with TMB substrate solution, and after color development for 20 minutes, 50 μL of 2 mol / L H2SO4 aqueous solution was added to terminate the reaction, and detected with an enzyme-labeled instrument, and the concentration of cytokines in each well plate was calculated by the standard curve established by the standard provided by the kit.

[0090] Table 6 Proliferation activity ratio of chondrocytes and synovial cells (%, n=6)

[0091]

[0092] Note: *P<0.05, **P<0.01 compared with the control group; △P<0.05, △△P<0.01 compared with the model group.

[0093] Table 7 Serum RANKL and OPG expression levels of rats in each group ( n=10)

[0094]

[0095] Note: *P<0.05, **P<0.01 compared with the control group; △P<0.05, △△P<0.01 compared with the model group.

[0096] From the results in Tables 6 and 7, it can be seen that the combination of the Chinese medicine preparation of the present invention and methotrexate can inhibit the osteoclast function of adjuvant-induced RA rats through the OPG / RANKL / RANK signaling pathway, and at the same time has the therapeutic effect of inhibiting cartilage loss, promoting cartilage repair, and inhibiting synovial hyperplasia.

[0097] Example 2 Toxicological Test of Combined Drugs

[0098] Twenty SPF SD rats, half male and half female, weighing 180-220g, were randomly divided into two groups according to sex and weight: a Chinese medicine preparation group (dose was 1.5 times the normal dosage) and a control group. The rats were fasted but not watered for 12 hours before the experiment. On the second day, the Chinese medicine compound preparation group was gavaged with 22.5g of raw medicine / kg, and the control group was gavaged with an equal volume of normal saline at 9:00, 12:00 and 17:00 respectively. The animal reactions were observed immediately after administration, and the appearance, behavioral activities, mental state, including breathing, changes in the nature and frequency of activities, convulsions, involuntary or voluntary muscle contractions, reflexes, symptoms of eye toxicity, salivation, fur, pain, gastrointestinal symptoms (feces, vomiting) and death were recorded. The observation was continued for 14 days, and the weight gain of the animals in each group was recorded before administration and 3, 7, and 14 days after administration. The results are shown in Tables 8 and 9.

[0099] Blood samples of rats in each group in the anti-inflammatory test of 1.2 were taken to prepare serum, and the serum AST and ALT levels were determined using serum aspartate aminotransferase (AST) and alanine aminotransferase (ALT) detection kits. The specific operation steps were strictly carried out according to the instructions. The results are shown in Table 10.

[0100] Table 8 Toxicity test body weight observation results (g, n=10)

[0101]

[0102] Note: *P<0.05 compared with the control group.

[0103] Table 9 Adverse reaction observation results (n=10)

[0104]

[0105]

[0106] Table 10 Changes in liver function index levels ( n=10)

[0107] Group AST ALT Blank control group 38.04±8.27 133.14±14.52 Chinese medicine preparation group 40.32±7.76 135.70±13.18 Methotrexate group <![CDATA[90.68±13.49 * ]]> <![CDATA[158.33±16.06 * ]]> Combination group 41.45±10.53 139.76±11.25

[0108] Note: *P<0.05 compared with the control group.

[0109] The results showed that there was no significant difference in weight gain compared with the control group (P>0.05). The appearance, behavioral activities and mental state of the animals were observed for 14 consecutive days. Except for one rat in the methotrexate group that had adverse reactions of weight loss and slow movement, no abnormalities were found in the animals in other groups. On the 14th day, some rats (including rats with adverse reactions) were killed and dissected. No obvious abnormalities were found in the main organs by naked eye observation. It can be seen that the Chinese medicine preparation disclosed in the present invention has few adverse reactions and good safety.

[0110] The changes in the levels of animal liver function indicators showed that when the Chinese medicine preparation of the present invention was used in combination with methotrexate, the levels of liver function indicators ALT and AST were normal, while the levels of the two transaminases increased in the rats in the methotrexate group, proving that the Chinese medicine preparation of the present invention can be used in combination with methotrexate to improve the therapeutic effect while reducing the dosage of methotrexate, thereby reducing the effect of methotrexate on the liver function of RA rats.

[0111] Example 3 Clinical trial of combined drug treatment of rheumatoid arthritis

[0112] 3.1 General Information

[0113] Inclusion criteria: ① Meet the diagnosis criteria of the "Diagnostic Criteria for Rheumatoid Arthritis" established by the American College of Rheumatology (ACR); ② Meet the relevant diagnostic criteria in the "Practical Chinese Medicine Rheumatology"; ③ 3 or more joint soft tissue swelling and morning stiffness lasting more than 1 hour; ④ Age between 45 and 80 years old; ⑤ Good compliance and voluntary participation in this study; ⑥ Complete clinical data.

[0114] Exclusion criteria: ① severe liver, heart, kidney and other functional disorders; ② taking immunosuppressants, hormones or drugs that affect bone metabolism in the past two months; ③ allergic to the study drugs or their ingredients; ④ being pregnant or lactating; ⑤ having severe bone hyperplasia or gastrointestinal functional diseases; ⑥ accompanied by other rheumatic autoimmune diseases.

[0115] 72 patients with rheumatoid arthritis were randomly divided into a control group and a combined medication group (36 cases in each group) according to the random number table method. The age of the patients in the control group (12 males and 24 females) ranged from 46 to 78 years old, with an average age of (62.81±8.56) years; their disease course ranged from 2 to 22 years, with an average disease course of (12.03±4.98) years. The age of the patients in the combined medication group (13 males and 23 females) ranged from 45 to 79 years old, with an average age of (60.72±9.01) years; their disease course ranged from 2 to 23 years, with an average disease course of (11.39±5.67) years. There was no significant difference in the general data of the two groups, such as gender, age and disease course (P>0.05), and they were comparable.

[0116] 3.2 Treatment methods

[0117] The patients in the control group were treated with methotrexate tablets (Shanghai SPH Xinyi Pharmaceutical Co., Ltd., National Medicine Standard H31020644), taken orally, twice a week, 10 mg each time; the patients in the combined medication group were treated with Chinese medicine preparations combined with methotrexate tablets, taken orally, 6 g of Chinese medicine preparations each time, twice a day, and 5 mg of methotrexate tablets each time, twice a week. The course of treatment was 12 weeks, and all 72 patients in both groups completed the treatment.

[0118] 3.3 Observation indicators

[0119] 3.3.1 Safety indicators: blood and urine routine tests, liver and kidney function. Record once a month.

[0120] 3.3.2 Efficacy indicators: number of joint tenderness, number of joint swelling, morning stiffness duration, C-reactive protein (CRP), erythrocyte sedimentation rate (ESR) and pain score assessed by visual analogue scale (VAS) (score 0-10 points: no pain 0 points; mild pain 1-3 points; moderate pain 4-6 points, affecting sleep, tolerable; severe pain 7-10 points, both sleep and appetite are affected, difficult to tolerate). Record once before and after treatment.

[0121] 3.3.3 Evaluation of efficacy The evaluation criteria were formulated by ACR and divided into three levels: clinically significant effect, therapeutically effective, and ineffective. The therapeutic effect was evaluated based on the number of joint tenderness and swelling and the main symptom score. Significant effect: the number of joint tenderness and swelling decreased by ≥50%, and the main symptom score decreased by ≥70%; effective: the number of joint tenderness and swelling decreased by ≥30%, and the main symptom score decreased by ≥40%; ineffective: the above standards were not met. Total effective rate = (number of significant effect cases + number of effective cases) / total number of cases × 100%.

[0122] 3.4 Statistical analysis

[0123] Statistical analysis was performed using SPSS 21.0 statistical software. The experimental quantitative data were expressed as mean ± standard deviation. Repeated measures analysis of variance was used for comparison among multiple groups. The LSD-t method was used for test between two groups. P < 0.05 indicated a significant statistical difference.

[0124] 3.5 Results display

[0125] 3.5.1 Comparison of treatment effects

[0126] Compared with the total effective rate of treatment, the combined medication group was significantly higher than the control group (P<0.05), see Table 11.

[0127] Table 11 Comparison of therapeutic effects between two groups of patients with rheumatoid arthritis [cases (%)]

[0128] Group Number of cases Significant effect efficient invalid Total Effectiveness Control group 36 8(22.2) 17(47.2) 11(30.6) 25(69.4) Combination medication group 36 12(33.3) 19(52.8) 5(13.9) <![CDATA[31(86.1) * ]]>

[0129] Note: *P<0.05 compared with the control group.

[0130] 3.5.2 Comparison of treatment efficacy indicators

[0131] After treatment, the number of swollen joints, tenderness, morning stiffness time, VAS score and CRP level in both groups were significantly reduced, and those in the combined medication group were significantly lower than those in the control group (P<0.05), as shown in Table 12.

[0132] Table 12 Comparison of joint swelling, tenderness and VAS scores between the two groups of patients with rheumatoid arthritis ( n=36)

[0133]

[0134] Note: *P<0.05 compared with before treatment, △P<0.05 compared with control group after treatment.

[0135] 3.5.3 Comparison of safety indicators

[0136] The levels of ALT, AST, serum creatinine (Scr) and blood urea nitrogen (BUN) were compared between the control group and the combination group before and after treatment. The results showed no statistical difference (P>0.05), as shown in Table 13.

[0137] Table 13 Comparison of ALT, AST, Scr, and BUN levels between the two groups of patients with rheumatoid arthritis (x ± s, n = 36)

[0138]

[0139] From the above results, it can be seen that after the treatment, the efficacy of the combined medication group was 86.1%, and the treatment effects in terms of joint swelling, tenderness, morning stiffness time, VAS score, and CRP level were significantly better than those in the control group (P<0.05). Therefore, it is proved that the Chinese medicine preparation disclosed in the present invention can cooperate with methotrexate to treat RA and achieve better therapeutic effects.

[0140] Finally, it should be noted that the above embodiments are used to illustrate the technical solution of the present disclosure rather than to limit the protection scope of the present disclosure. Although the present disclosure has been described in detail with reference to the preferred embodiments, those skilled in the art should understand that the technical solution of the present disclosure can be modified or replaced by equivalents without departing from the essence and scope of the technical solution of the present disclosure.

Claims

1. A combined drug for treating rheumatoid arthritis, characterized in that: include: (a) a Chinese medicine composition containing the following raw materials in a therapeutically effective amount: tortoise shell glue, antler glue, Morinda officinalis, psoralea corylifolia, Dipsacus asper, Eucommia ulmoides, Rehmannia root, Angelica sinensis, White peony root, Lycium barbarum, Schisandra chinensis, Chinese yam, Cornus officinalis, Ophiopogon japonicus, Euryale ferox, Cinnamon bark, Aconite root, Paeonia suffruticosa moutan, Alisma orientalis, Poria, and, optionally, a pharmaceutically acceptable carrier or excipient; (b) a therapeutically effective amount of methotrexate or a pharmaceutically acceptable derivative thereof, and, optionally, a pharmaceutically acceptable carrier or excipient.

2. The combined drug according to claim 1, characterized in that: The traditional Chinese medicine composition comprises the following raw materials in parts by weight: 1-5 parts of tortoise shell glue, 1-5 parts of antler glue, 5-10 parts of Morinda officinalis, 2-8 parts of psoralea corylifolia, 5-10 parts of Dipsacus asper, 5-10 parts of Eucommia ulmoides, 10-15 parts of Rehmannia root, 5-10 parts of Angelica sinensis, 5-10 parts of white peony root, 2-8 parts of wolfberry, 1-5 parts of Schisandra chinensis, 10-20 parts of yam, 5-15 parts of Cornus officinalis, 2-8 parts of Ophiopogon japonicus, 10-15 parts of Euryale ferox, 1-5 parts of cinnamon bark, 2-8 parts of aconite, 5-15 parts of Paeonia suffruticosa, 5-15 parts of Alisma orientalis, and 5-15 parts of Poria.

3. The combined drug according to claim 1, characterized in that: The tortoise shell glue and deer antler glue need to be melted.

4. Use of the combined drug according to any one of claims 1 to 3 in the preparation of a drug for treating rheumatoid arthritis.

5. A medicine kit for treating rheumatoid arthritis, characterized in that: The medicine kit comprises at least one first preparation unit and at least one second preparation unit, and the first preparation unit and the second preparation unit are both independently present in the medicine kit; The first preparation unit comprises a therapeutically effective amount of a Chinese medicine composition containing the following raw materials: tortoise shell glue, deer horn glue, Morinda officinalis, psoralea corylifolia, Dipsacus asper, Eucommia ulmoides, Rehmannia root, Angelica sinensis, White peony root, Lycium barbarum, Schisandra chinensis, Chinese yam, Cornus officinalis, Ophiopogon japonicus, Euryale ferox, Cinnamon bark, Aconite root, Paeonia suffruticosa moutan, Alisma orientalis, Poria cocos, and, optionally, a pharmaceutically acceptable carrier or excipient; The second dosage unit comprises a therapeutically effective amount of methotrexate or a pharmaceutically acceptable derivative thereof, and, optionally, a pharmaceutically acceptable carrier or excipient.

6. The medicine box according to claim 5, characterized in that The traditional Chinese medicine composition comprises the following raw materials in parts by weight: 1-5 parts of tortoise shell glue, 1-5 parts of antler glue, 5-10 parts of Morinda officinalis, 2-8 parts of psoralea corylifolia, 5-10 parts of Dipsacus asper, 5-10 parts of Eucommia ulmoides, 10-15 parts of Rehmannia root, 5-10 parts of Angelica sinensis, 5-10 parts of white peony root, 2-8 parts of wolfberry, 1-5 parts of Schisandra chinensis, 10-20 parts of yam, 5-15 parts of Cornus officinalis, 2-8 parts of Ophiopogon japonicus, 10-15 parts of Euryale ferox, 1-5 parts of cinnamon bark, 2-8 parts of aconite, 5-15 parts of Paeonia suffruticosa, 5-15 parts of Alisma orientalis, and 5-15 parts of Poria.

7. The combined drug according to any one of claims 1 to 2 or the drug kit according to any one of claims 5 to 6, characterized in that: The dosage form of the Chinese medicine composition includes one of tablets, powders, granules, pills, capsules, solutions, emulsions and injections.

8. The medicine box according to claim 5, characterized in that The medicine kit also includes instructions, which stipulate that the first preparation unit and the second preparation unit are used as preparations for combined use in the treatment of rheumatoid arthritis.

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