A composition for treating rheumatoid arthritis

Abstract

The invention provides a composition for treating rheumatoid arthritis, belonging to the technical field of traditional Chinese medicine preparations. Strychnine (from Strychnos nux vomica or Strychnos nux vomica extract), atractylodes lactone III (from Atractylodes macrocephala or Atractylodes macrocephala extract), tanshinone IIA (from Salvia miltiorrhiza or Salvia miltiorrhiza extract) and ligusticolide A (from Chuanxiong or Chuanxiong extract) are selected and compounded in a suitable ratio to achieve a good therapeutic effect on rheumatoid arthritis. The invention can achieve more accurate medication by accurately controlling the ratio of each effective ingredient, thereby ensuring the therapeutic effect, reducing the side effects that may be caused by excessive ingredients, and improving the safety of the drug and the tolerance of patients.

Description

Technical Field

[0001] The invention belongs to the technical field of traditional Chinese medicine preparations, and in particular relates to a composition for treating rheumatoid arthritis. Background Art

[0002] Rheumatoid arthritis (RA) is a chronic systemic autoimmune disease characterized by synovial inflammation, cartilage and bone destruction, which ultimately leads to joint deformity and loss of function. With the continuous advancement of medical technology, the treatment of RA has become increasingly abundant, but it still faces many challenges.

[0003] At present, drug therapy is the core of RA treatment. Commonly used drugs include nonsteroidal anti-inflammatory drugs (NSAIDs), glucocorticoids, immunosuppressants (such as methotrexate, leflunomide) and biologics. NSAIDs are used to relieve pain and inflammation, while glucocorticoids can quickly control acute symptoms, but long-term use has significant side effects. Immunosuppressants and biologics regulate the immune system, reduce inflammatory responses, and control disease progression. However, these drugs often require long-term or even lifelong use, and some patients do not respond well to the drugs or experience serious side effects.

[0004] With the development of biotechnology, biological agents have gradually occupied an important position in the treatment of RA. Biological agents specifically inhibit inflammatory cytokines such as tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6), thereby blocking the inflammatory process and reducing joint damage. Biological agents such as TNF-α antagonists, IL-6 antagonists, and CD20 inhibitors have shown the advantages of rapid onset and good efficacy in the treatment of RA, and have become an important choice for many patients. However, biological agents also have certain defects, such as increased risk of infection and tumors, and require strict evaluation and monitoring before use.

[0005] Physical therapy, such as heat therapy, cold therapy, massage, physical therapy, and exercise therapy, plays an auxiliary role in the treatment of RA. These methods can relieve joint pain and swelling, promote blood circulation, and improve joint flexibility and stability. However, the effect of physical therapy varies from person to person, and it cannot cure RA.

[0006] In recent years, new therapeutic drugs for RA have continued to emerge, such as JAK (Janus kinase) inhibitors, IL-17 inhibitors, etc. These new drugs provide more treatment options for RA patients through different mechanisms of action. In particular, JAK inhibitors are believed to have a wider range of anti-inflammatory effects because they can block the signal transduction of multiple inflammatory cytokines. However, their long-term safety and effectiveness still need to be further verified. Summary of the invention

[0007] The purpose of this section is to summarize some aspects of embodiments of the present invention and briefly introduce some preferred embodiments. Some simplifications or omissions may be made in this section and the specification abstract and the invention title of this application to avoid blurring the purpose of this section, the specification abstract and the invention title, and such simplifications or omissions cannot be used to limit the scope of the present invention.

[0008] In view of the above problems and / or the problems existing in the prior art, the present invention is proposed.

[0009] Therefore, the object of the present invention is to overcome the deficiencies in the prior art and provide a composition for treating rheumatoid arthritis.

[0010] In order to solve the above technical problems, the present invention provides the following technical solution: a Chinese herbal medicine or an extract thereof containing the following effective ingredients,

[0011] Strychnine-containing seeds of Strychnos nux vomica or their extracts, atractylodes macrocephala or their extracts containing atractylodes lactone III, salvia miltiorrhiza or their extracts containing tanshinone ⅡA, chuanxiong or their extracts containing chuanxiong lactone A;

[0012] Among them, the content ratio of strychnine to atractylodes lactone III, tanshinone ⅡA, and ligusticum chuanxiong lactone A is 200:1~5:2.5~10:30~100.

[0013] As a preferred embodiment of the composition for treating rheumatoid arthritis of the present invention, the composition further comprises licorice or licorice extract, Codonopsis pilosula or Codonopsis pilosula extract, Poria cocos or Poria cocos extract, Panax notoginseng or Panax notoginseng extract, Achyranthes bidentata or Achyranthes bidentata extract, and Pheretima lumbricoides or Pheretima lumbricoides extract.

[0014] As a preferred embodiment of the composition for treating rheumatoid arthritis of the present invention, the content ratio of strychnine to atractylodes lactone III is 200:1-2.5.

[0015] As a preferred embodiment of the composition for treating rheumatoid arthritis of the present invention, the content ratio of strychnine to tanshinone ⅡA is 200:2.5-5.

[0016] As a preferred embodiment of the composition for treating rheumatoid arthritis of the present invention, the content ratio of strychnine to ligusticum lactone A is 200:30-50.

[0017] As a preferred embodiment of the composition for treating rheumatoid arthritis of the present invention, the mass ratio of strychnine in the strychnine seed or strychnine seed extract to licorice or licorice extract, Codonopsis pilosula or Codonopsis pilosula extract, Poria cocos or Poria cocos extract, Panax notoginseng or Panax notoginseng extract, Achyranthes bidentata or Achyranthes bidentata extract, and Pheretima or Pheretima extract is 0.1:5~20:10~30:10~20:5~20:5~15:0.5~5.

[0018] As a preferred embodiment of the composition for treating rheumatoid arthritis of the present invention, the mass ratio of strychnine in the strychnine seed or strychnine seed extract to licorice or licorice extract, Codonopsis pilosula or Codonopsis pilosula extract, and Poria cocos or Poria cocos extract is 0.1:5~15:10~15:10~15.

[0019] As a preferred embodiment of the composition for treating rheumatoid arthritis of the present invention, the mass ratio of strychnine in the Strychnos nux vomica or Strychnos nux vomica extract to Panax notoginseng or Panax notoginseng extract and Achyranthes bidentata or Achyranthes bidentata extract is 0.1:10~20:5~10.

[0020] As a preferred embodiment of the composition for treating rheumatoid arthritis of the present invention, the mass ratio of strychnine in the strychnine or strychnine extract to earthworm or earthworm extract is 0.1:1-5.

[0021] Another object of the present invention is to provide a composition for use in preparing a drug for treating rheumatoid arthritis.

[0022] Beneficial effects of the present invention:

[0023] The present invention can achieve more accurate medication by precisely controlling the ratio of each effective ingredient, which not only ensures the therapeutic effect, but also reduces the side effects that may be caused by excessive ingredients, thereby improving the safety of the drug and the tolerance of patients. DETAILED DESCRIPTION

[0024] In order to make the above-mentioned objects, features and advantages of the present invention more obvious and easy to understand, the specific implementation methods of the present invention are described in detail below in conjunction with the embodiments of the specification.

[0025] In the following description, many specific details are set forth to facilitate a full understanding of the present invention, but the present invention may also be implemented in other ways different from those described herein, and those skilled in the art may make similar generalizations without violating the connotation of the present invention. Therefore, the present invention is not limited to the specific embodiments disclosed below.

[0026] Secondly, the term "one embodiment" or "embodiment" as used herein refers to a specific feature, structure, or characteristic that may be included in at least one implementation of the present invention. The term "in one embodiment" that appears in different places in this specification does not necessarily refer to the same embodiment, nor does it refer to a separate or selective embodiment that is mutually exclusive with other embodiments.

[0027] The method for obtaining the active substance extract corresponding to the Chinese medicinal materials used in the specific embodiment of the present invention is as follows:

[0028] Preparation method of Nux Vomica extract:

[0029] Crush the Strychnos nux vomica and add an equal amount of methylamine solution to soak for 3 hours, then add 5 times the amount of ethyl acetate, reflux and extract twice, each time for 9 hours, filter and combine the filtrates, concentrate under reduced pressure to 1 / 3 of the original extract volume, and filter to obtain the extract;

[0030] The extract was added to a silica gel chromatography column, eluted with ethanol, and the eluate after 5 times the column volume was collected. The eluate was concentrated and dried, and crushed to obtain a crushed product;

[0031] The crushed material was added with 35 times the amount of 1% sulfuric acid and heated to dissolve, filtered to obtain filtrate C, potassium hydroxide solution was added to filtrate C to adjust the pH of the filtrate to 10, allowed to stand at room temperature, filtered, and dried to obtain powder;

[0032] The powder was added with 20 times amount of 30% ethanol, heated and dissolved, filtered, the filtrate was crystallized, filtered, the filter cake was dried and crushed to obtain the strychnine extract, and the strychnine content was 97% after determination.

[0033] Preparation method of Atractylodes macrocephala extract:

[0034] The Atractylodes macrocephala was crushed and passed through a 10-mesh sieve to obtain Atractylodes macrocephala powder, and the Atractylodes macrocephala powder was soaked in 20% by volume hydrogen peroxide for 4 hours, and then oxidized at 50° C. for 8 hours to obtain oxidized Atractylodes macrocephala powder;

[0035] The oxidized Atractylodes macrocephala powder was dried at 40°C for 12 hours to obtain dry Atractylodes macrocephala powder;

[0036] The dried Atractylodes macrocephala powder was added to 95% ethanol for reflux extraction for 1 hour, and zinc particles of 5% of the mass of the dried Atractylodes macrocephala powder were added before reflux extraction to obtain the Atractylodes macrocephala total lactone extract;

[0037] The total lactone extract of Atractylodes macrocephala is concentrated under reduced pressure to obtain a concentrated solution, which is then freeze-dried to obtain the Atractylodes macrocephala lactone extract. It is determined that the content of Atractylodes macrocephala lactone III is 20%.

[0038] Preparation method of Danshen extract:

[0039] The powder of Danshen was obtained by crushing Danshen, and then soaked in 95% ethanol for 1 hour, and then stored at 35kHz and 40℃ for 30 minutes, and the filtrate was filtered to obtain the filtrate, and then concentrated and dried under reduced pressure to obtain the dry extract;

[0040] Ethyl acetate and sodium bicarbonate solution were fully mixed in a volume ratio of 7:10, and the layer was allowed to stand to seal. The upper solution and the lower solution were mixed in a volume ratio of 2:1 to obtain a mixed solution.

[0041] The dry extract was dissolved in the mixed solution and extracted in the dark at room temperature for 1 h to obtain an ethyl acetate layer, to which anhydrous sodium sulfate was added for drying, and the layer was placed at 4°C for 6 h. The anhydrous sodium sulfate was removed by filtration, and the ethyl acetate was evaporated in a 50°C water bath to obtain a Salvia miltiorrhiza extract. The content of cryptotanshinone in the extract was determined to be 5%.

[0042] Preparation method of Chuanxiong extract:

[0043] Crush Chuanxiong to obtain Chuanxiong powder, add the Chuanxiong powder to 10 times the amount of 75% ethanol solution, soak overnight at room temperature, extract at 40°C for 1h, filter, repeat twice, combine the filtrate, recover ethanol under reduced pressure at 50°C to obtain an extract;

[0044] Add 1 volume of distilled water to the extract, mix well, then add 5 volumes of extraction solvent (ethyl acetate and petroleum ether mixed in a ratio of 5:1) of the extract, stir under pressure at room temperature for 10 min, let stand for 24 h, separate the organic layer, recover the solvent under reduced pressure to obtain Chuanxiong extract, and the content of ligusticum chuanxiong lactone A in it was determined to be 10%.

[0045] Efficacy test (constructing a rat rheumatoid arthritis model to examine the efficacy of each composition on rats with rheumatoid arthritis)

[0046] 1. Materials

[0047] 1.1 Drugs and reagents

[0048] The compositions of various embodiments / comparative examples;

[0049] Prednisone acetate tablets: Xinxiang Changle Pharmaceutical Co., Ltd., specification 5 mg / tablet, batch number 21020162;

[0050] Leigongzong Tablets: Shanghai Fudan Fuhua Pharmaceutical Co., Ltd., specification 10 mg / tablet, batch number 180502;

[0051] Chicken type II collagen, Chondrex: product number 20012;

[0052] Rat interleukin-1β kit (ELISA method): R&D Systems, batch number P261453;

[0053] Rat interleukin-17 kit (ELISA method): Shanghai Xitang Biotechnology Co., Ltd., batch number 2104251;

[0054] Rat interleukin-10 kit (ELISA method): ExCellBio, batch number 22J225;

[0055] Rat tumor necrosis factor-a kit (ELISA method): R&D Systems batch number P254162;

[0056] Rheumatoid factor (RF) assay kit: Nanjing Jiancheng Bioengineering Institute, batch number 20210407;

[0057] Rat interferon-γ kit (IFN-γ): Biolegend Company, batch number B303835.

[0058] 1.2 Main test instruments

[0059] YLS-7B foot swelling instrument: Equipment Station of Shandong Academy of Medical Sciences;

[0060] STXI23ZH electronic balance: Ohaus Instruments (Changzhou) Co., Ltd.;

[0061] SpectraMaxM5 microplate reader: Molecular Devices, USA;

[0062] WD-2103B plate washer: Beijing Liuyi Biotechnology Co., Ltd.;

[0063] SorallST8R high-speed refrigerated centrifuge: Thero, Germany;

[0064] HPX-9052MBE electric constant temperature incubator: Shanghai Boxun Industrial Co., Ltd.;

[0065] DW-30L508 Medical cryogenic storage box: Haier Biomedical;

[0066] ASP200S automatic vacuum tissue dehydrator, EG1150H automatic biological tissue embedding machine: Leica, Germany;

[0067] RM2255 slicer: Leica, Germany;

[0068] CiL microscope: Nikon Corporation, Japan.

[0069] 1.3 Experimental animals

[0070] Rats: strain SD, raised in an experimental animal barrier system, temperature, humidity, and ventilation times are automatically controlled by the central system, the temperature is maintained at 20-26°C, the relative humidity is maintained at 40%-70%, the ventilation times are 10-15 times / h fresh air, and the lighting is 12h light and 12h dark. Free access to food and water, rat feed is provided by Beijing Keao Xieli Feed Co., Ltd., and drinking water is purified water (prepared by KFRO-400GPD water purifier).

[0071] 2. Methods

[0072] 2.1 Model preparation

[0073] (1) Preparation of type II collagen suspension emulsion: 10 mL of 0.1 mol / L acetic acid solution and 20 mg of chicken type II collagen were mixed in a brown reagent bottle under ice bath until the white chicken type II collagen was completely dissolved in the acetic acid. At this time, the concentration of chicken type II collagen in the obtained solution was 2 mg / mL. Before the experiment, Freund's complete adjuvant was added and fully emulsified (the most critical step in the emulsion preparation process, whether the emulsification is sufficient directly affects the success of the CIA model establishment) to make the chicken type II collagen concentration 1 mg / mL.

[0074] (2) Preparation of CIA rat model: 0.1 mL of chicken type II collagen was injected intradermally at the left foot pad. After injection, the injection point was pressed to prevent liquid outflow. Rats in the normal control group were injected with an equal volume of saline solution in the same way. On the 7th day after the first injection, 0.1 mL of the same emulsion was injected into the back for booster immunization. The rats were closely observed until 15 days after modeling, and animals with inflammation and swelling in bilateral joints were selected for grouping.

[0075] 2.2 Grouping

[0076] Several male SD rats weighing 110-130g were fed for one week during the adaptation period. Ten rats were randomly selected as the normal control group, and all rats were subjected to chicken type II collagen modeling. The rats with successful modeling were selected and re-marked and randomly divided into model group, prednisone group (PDN), total glycosides of tripterygium wilfordii group (TG), and (BQ) experimental drug group.

[0077] 2.3 Administration

[0078] After successful modeling, the animals were divided into groups. The prednisone group was given 10 mg / kg by oral gavage, the triptolide group was given 10 mg / kg by oral gavage, each medication group (example and comparative example) was given 0.2 g / kg by oral gavage, and the blank group and model group were given normal saline for 15 consecutive days, during which they were given regular feed and drinking water for SPF animals.

[0079] 2.4 Observation indicators

[0080] 2.4.1 Body weight, tissue swelling, and arthritis score

[0081] Body weight: The rats were weighed on the 1st, 5th, 10th and 15th day after the start of group administration, and the average body weight of each group of rats was calculated.

[0082] Foot swelling: The swelling of the right hind paw of rats was measured with a foot swelling meter on days 1, 5, 10 and 15 after the start of group administration, and the average swelling of the left hind paw of rats in each group was counted.

[0083] Arthritis score: 14 days after the injection of type II collagen in the left hind paw, secondary arthritis will form in the right hind paw. The arthritis index score will be monitored on the 1st, 5th, 10th and 15th days after group administration. The arthritis index score is determined by the following scoring system: 0 points represent normal paws; 0-1 points represent redness and swelling of toes; 1-2 points represent redness and swelling of toes and soles; 2-3 points represent swelling below the ankle joint; 3-4 points represent complete swelling of the entire paw and inability to bend. The scores of the four paws of each rat are added together as the joint score, and the maximum score for each rat is 16.

[0084] 2.4.2 Serum rheumatoid factor

[0085] The subjects were fasted for more than 12 h before the last administration and were allowed to drink water freely. One hour after the last administration, blood was collected from the abdominal aorta after anesthesia with sodium pentobarbital in a blood collection tube. The blood was centrifuged at 3000 rpm and 4°C for 10 min to separate the plasma samples, which were stored at -80°C for future use in the detection of rheumatoid factor (RF).

[0086] 2.4.3 Serum inflammatory factors

[0087] The -80℃ frozen serum was reconstituted and the levels of IL-17, L-10, TNF-0, ⅡL-Iβ and IFN-γ were detected by ELISA.

[0088] 2.4.4 Ankle joint histopathological examination

[0089] The right ankle joint of the rats was cut off and fixed with 10% paraformaldehyde solution. The tissue was treated with formic acid formaldehyde decalcification solution, dehydrated with different concentrations of ethanol solution, transparentized with xylene, embedded in stone, sliced, and stained with hematoxylin and eosin in sequence. After sealing, the synovial hyperplasia, inflammatory cell infiltration, joint cavity leaching, articular cartilage destruction, fibrosis, etc. of the ankle joints of each group were observed and compared under a microscope.

[0090] 2.5 Statistical methods

[0091] All data are expressed as mean ± standard error ( +sem) were used to analyze the differences among the groups, and P < 0.05 indicated that the differences among the groups were statistically significant.

[0092] Example 1

[0093] This embodiment provides a composition for treating rheumatoid arthritis, specifically:

[0094] Weigh the extract of Strychnos nux vomica, the extract of Atractylodes macrocephala, the extract of Salvia miltiorrhiza, and the extract of Ligusticum chuanxiong, so that the content ratio of strychnine to atractylodes lactone III, tanshinone ⅡA, and ligustrin lactone A is 200:2.5:5:50;

[0095] The above active ingredients are ground into powder and mixed to obtain the composition of this embodiment.

[0096] Example 2

[0097] Weigh the extract of Strychnos nux vomica, the extract of Atractylodes macrocephala, the extract of Salvia miltiorrhiza, and the extract of Ligusticum chuanxiong so that the content ratio of brucine to atractylodes lactone III, tanshinone ⅡA, and ligusticum chuanxiong lactone A is 200:2.5:5:50;

[0098] Weigh licorice, codonopsis, tuckahoe, panax notoginseng, achyranthes bidentata, and earthworm, so that the mass ratio of strychnine in the strychnine extract to licorice, codonopsis, tuckahoe, panax notoginseng, achyranthes bidentata, and earthworm is 0.1:15:15:15:10:10:1;

[0099] The above active ingredients are ground into powder and mixed to obtain the composition of this embodiment.

[0100] The pharmacodynamic effects of the compositions of Example 1 and Example 2 on rats with rheumatoid arthritis were investigated according to the above method. The results are shown in Tables 1 and 2.

[0101] Table 1 Effects of different compositions on body weight, foot swelling and arthritis scores in rats with rheumatoid arthritis

[0102]

[0103] (Note: Compared with the normal group, & p<0.05, && p<0.01; compared with the model group, * p<0.05, ** p<0.01)

[0104] As shown in Table 1, after type II collagen modeling, the rats lost weight, and their toes became red and swollen, their joints swollen, and their joints deformed, making it difficult for them to walk. In severe cases, they even lost the ability to move, indicating that a rheumatoid arthritis rat model was successfully established. The body weight, tissue swelling, and arthritis scores of the rats were tested during continuous drug administration. The results showed that as the drug administration time increased, the body weight and arthritis scores of the arthritis rats in each drug administration group were improved.

[0105] After 5 days of administration, the compositions of Examples 1 and 2 of the present invention both showed significant reduction in the swelling of the toes of rats. After 10 days of administration, all groups showed significant reduction in the swelling of the toes of rats, and the compositions of Examples 1 and 2 of the present invention were superior to the prednisone group and the total glycosides of Tripterygium wilfordii group.

[0106] In summary, the compositions of Examples 1 and 2 of the present invention can improve the state of arthritis rats and have obvious anti-inflammatory effects, which are manifested in improving the swelling of toes and reducing the arthritis score.

[0107] Table 2 Effects of different compositions on serum rheumatoid factors and inflammatory factors in rats with rheumatoid arthritis

[0108] Group dose RF (IU / ml) IL-17 (pg / ml) IFN-γ (pg / ml) TNF-α (pg / ml) IL-1β (pg / ml) IL-10 (pg / ml) Normal group - 140.87±3.64 61.13±8.41 8.50±0.67 5.81±0.31 18.70±2.47 27.03±10.04 Model Group - <![CDATA[180.23±4.83 && ]]> <![CDATA[94.67±8.95 & ]]> <![CDATA[47.95±15.21 && ]]> <![CDATA[11.56±2.08 && ]]> <![CDATA[39.60±6.21 && ]]> 15.01±3.92 Prednisone group 10mg / kg <![CDATA[146.07±3.15 ** ]]> <![CDATA[64.81±5.15 * ]]> <![CDATA[13.40±2.14 ** ]]> <![CDATA[6.06±0.36 *** ]]> <![CDATA[21.25±2.41 ** ]]> 26.74±3.86 Total glycosides of Tripterygium wilfordii 10mg / kg <![CDATA[164.93±2.48 * ]]> <![CDATA[52.96±7.47 ** ]]> <![CDATA[9.67±0.43 ** ]]> <![CDATA[6.62±0.47 ** ]]> <![CDATA[16.27±2.35 *** ]]> 18.42±5.13 Example 1 0.2g / kg <![CDATA[141.92±4.42 *** ]]> <![CDATA[51.96±6.65 *** ]]> <![CDATA[8.81±0.56 ** ]]> <![CDATA[5.53±1.21 *** ]]> <![CDATA[17.72±2.18 *** ]]> 26.01±2.65 Example 2 0.2g / kg <![CDATA[140.36±4.65 *** ]]> <![CDATA[49.02±6,87 *** ]]> <![CDATA[8.79±3.78 ** ]]> <![CDATA[5.56±3.21 *** ]]> <![CDATA[17.97±1.25 *** ]]> 26.52±8.12

[0109] (Note: Compared with the normal group, & p<0.05, && p<0.01; compared with the model group, * p<0.05, ** p<0.01, *** p<0.001)

[0110] Cytokines play an important role in rheumatoid arthritis inflammation and joint damage. It can be seen from Table 2 that compared with the blank group, the inflammatory factors such as RF, IL-17, IFN-γ, TNF-α, IL-1β in the serum of the model group rats all showed increased levels. Compared with the model group, each administration group can significantly reduce the content of inflammatory factors such as RF, IL-17, IL-1β, and the compositions of Examples 1 and 2 of the present invention are superior to the prednisone and total glycosides of tripterygium wilfordii groups.

[0111] IL-10 has a strong anti-inflammatory effect. Increasing the content of IL-10 is beneficial to inhibiting the release and clearance of inflammatory factors in the body. The results showed that compared with the blank group, the IL-10 content in the serum of the model group showed a downward trend, while the drug-treated group showed the effect of improving the level of IL-10 in the body.

[0112] In summary, the compositions of Examples 1 and 2 of the present invention can effectively improve various symptoms of rheumatoid arthritis, and the effects of the two compositions are basically equivalent.

[0113] Comparative Example 1

[0114] The difference between this comparative example and Example 2 is that the Nux Vomica extract in the active content is omitted, and the rest of the formula is the same as Example 2 to obtain the composition of this comparative example.

[0115] Comparative Example 2

[0116] The difference between this comparative example and Example 2 is that the Atractylodes macrocephala extract in the active content is omitted, and the rest of the formula is the same as that in Example 2 to obtain the composition of this comparative example.

[0117] Comparative Example 3

[0118] The difference between this comparative example and Example 2 is that the salvia miltiorrhiza extract in the active content is omitted, and the rest of the formula is the same as that of Example 2 to obtain the composition of this comparative example.

[0119] Comparative Example 4

[0120] The difference between this comparative example and Example 2 is that the Chuanxiong extract in the active content is omitted, and the rest of the formula is the same as Example 2 to obtain the composition of this comparative example.

[0121] Comparative Example 5

[0122] The difference between this comparative example and Example 2 is that the licorice extract in the active content is omitted, and the rest of the formula is the same as Example 2 to obtain the composition of this comparative example.

[0123] Comparative Example 6

[0124] The difference between this comparative example and Example 2 is that the Codonopsis pilosula extract in the active content is omitted, and the rest of the formula is the same as that of Example 2 to obtain the composition of this comparative example.

[0125] Comparative Example 7

[0126] The difference between this comparative example and Example 2 is that the Poria cocos extract in the active content is omitted, and the rest of the formula is the same as Example 2 to obtain the composition of this comparative example.

[0127] Comparative Example 8

[0128] The difference between this comparative example and Example 2 is that the Panax notoginseng extract in the active content is omitted, and the rest of the formula is the same as Example 2 to obtain the composition of this comparative example.

[0129] Comparative Example 9

[0130] The difference between this comparative example and Example 2 is that the Achyranthes bidentata extract in the active content is omitted, and the rest of the formula is the same as that of Example 2 to obtain the composition of this comparative example.

[0131] Comparative Example 10

[0132] The difference between this comparative example and Example 2 is that the Achyranthes bidentata extract in the active content is omitted, and the rest of the formula is the same as that of Example 2 to obtain the composition of this comparative example.

[0133] The pharmacodynamic effects of the compositions of Comparative Examples 1 to 10 on rats with rheumatoid arthritis were investigated by referring to the above method and compared with Example 2. The results are shown in Table 3.

[0134] Table 3 Effects of different compositions on serum rheumatoid factors and inflammatory factors in rats with rheumatoid arthritis

[0135] Group RF (IU / ml) IL-17 (pg / ml) IFN-γ (pg / ml) TNF-α (pg / ml) IL-1β (pg / ml) IL-10 (pg / ml) Example 2 140.36±4.65 49.02±6,87 8.79±3.78 5.56±3.21 17.97±1.25 26.52±8.12 Comparative Example 1 <![CDATA[178.23±3.21 * ]]> <![CDATA[90.21±0.98 * ]]> <![CDATA[36.23±2.34 * ]]> <![CDATA[10.96±5.43 * ]]> <![CDATA[29.23±7.90 * ]]> 19.78±9.76 Comparative Example 2 <![CDATA[166.21±2.32 * ]]> <![CDATA[88.76±8.78 * ]]> <![CDATA[41.32±5.65 * ]]> <![CDATA[9.88±6.54 * ]]> <![CDATA[27.88±4.87 * ]]> 20.67±3.49 Comparative Example 3 <![CDATA[170.91±2.87 * ]]> <![CDATA[82.78±5.65 * ]]> <![CDATA[33.26±4.60 * ]]> <![CDATA[9.04±5.87 * ]]> <![CDATA[23.63±3.12 * ]]> 19.22±6.09 Comparative Example 4 <![CDATA[175.09±0.12 * ]]> <![CDATA[86.54±5.09 * ]]> <![CDATA[29.98±5.75 * ]]> <![CDATA[8.77±3.09 * ]]> <![CDATA[24.56±2.87 * ]]> 21.23±7.02 Comparative Example 5 145.56±3.45 54.23±3.56 12.67±3.78 6.54±5.06 18.99±4.98 24.01±5.67 Comparative Example 6 148.93±8.11 56.45±6.54 18.97±0.98 6.98±4.89 20.12±4.07 24.89±4.98 Comparative Example 7 151.45±2.54 59.34±3.67 19.88±5.89 7.45±3.96 21.22±5.62 25.55±5.55 Comparative Example 8 156.11±3.56 61.20±4.65 16.23±5.76 7.52±3.12 18.78±4.09 23.89±7.98 Comparative Example 9 149.92±9.21 54.38±0.67 23.36±4.08 5.97±0.98 19.23±0.95 24.67±4.09 Comparative Example 10 153.24±0.12 57.83±8.11 19.99±0.48 6.23±7.65 20.21±5.46 25.09±6.27

[0136] (Note: Compared with Example 2, * p<0.05)

[0137] As can be seen from Table 3, after omitting the extracts of Comparative Examples 1 to 4 in the scheme of the present invention, the improvement effect of the obtained composition on rheumatoid arthritis is significantly reduced compared with Example 2, and after omitting the extractions of Comparative Examples 5 to 10, the performance of the obtained composition is reduced compared with Example 2, but the reduction is not obvious. It can be inferred that in the scheme of the present invention, the extracts of Comparative Examples 1 to 4 play a key role in improving rheumatoid arthritis.

[0138] Example 3

[0139] This example is used to explore the effects of different ratios of Atractylodes lactone III to strychnine, tanshinone ⅡA, and ligusticum chuanxiong lactone A on the performance of the composition in treating rheumatoid arthritis. Specifically, the content ratios of strychnine to Atractylodes lactone III, tanshinone ⅡA, and ligusticum chuanxiong lactone A in Example 1 are adjusted to be: 200:0.5:5:50; 200:1:5:50;

[0142] 200:2.5:5:50 (Example 1); 200:5:5:50; 200:10:5:50;

[0145] The remaining steps and processes are all referred to Example 1 to obtain the compositions of this comparative example with different ratios of Atractylodes lactone III.

[0146] Referring to the above method, the effects of the compositions of different mass ratios of strychnine: atractylodes lactone III, tanshinone ⅡA, and ligusticum chuanxiong lactone A in this example on serum rheumatoid factor and inflammatory factors in rats with rheumatoid arthritis were compared. The results are shown in Table 4.

[0147] Table 4

[0148] Group RF (IU / ml) IL-17 (pg / ml) IFN-γ (pg / ml) TNF-α (pg / ml) IL-1β (pg / ml) IL-10 (pg / ml) Example 2 140.36±4.65 49.02±6.87 8.79±3.78 5.56±3.21 17.97±1.25 26.52±8.12 200：0.5：5：50 <![CDATA[187.67±0.87 * ]]> <![CDATA[85.32±23.3 * ]]> <![CDATA[38.81±7.90 * ]]> <![CDATA[10.10±9.87 * ]]> <![CDATA[30.12±1.19 * ]]> 20.23±8.01 200：1：5：50 150.23±8.90 59.32±4.66 15.20±5.28 7.43±6.09 21.23±2.09 23.63±3.14 200：2.5：5：50 141.92±5.34 51.96±7.09 8.81±1.19 5.53±5.43 17.72±3.15 26.01±1.16 200：5：5：50 148.96±8.45 64.21±2.09 10.23±2.01 6.91±3.29 23.36±4.50 23.65±3.7 200：10：5：50 <![CDATA[176.26±5.02 * ]]> <![CDATA[76.82±8.71 * ]]> <![CDATA[29.81±0.39 * ]]> <![CDATA[8.79±8.16 * ]]> <![CDATA[29.11±3.14 * ]]> <![CDATA[25.59±2.19 * ]]>

[0149] (Note: Compared with Example 2, * p<0.05)

[0150] As can be seen from Table 4, when the ratio of Atractylodes lactone III to strychnine is 1-5:200, its effect in improving rheumatoid arthritis is basically equivalent to that of Example 2, but when the ratio exceeds this range, its performance is significantly reduced compared with Example 2. This shows that within this range, there is a synergistic effect between the active compositions.

[0151] Example 4

[0152] This example is used to explore the effects of different ratios of tanshinone ⅡA to strychnine, atractylodes lactone Ⅲ, and ligusticum chuanxiong lactone A on the performance of the composition in treating rheumatoid arthritis. Specifically, the content ratios of strychnine to atractylodes lactone Ⅲ, tanshinone ⅡA, and ligusticum chuanxiong lactone A in Example 1 are adjusted to be: 200:2.5:1:50; 200:2.5:2.5:50;

[0155] 200:2.5:5:50 (Example 1); 200:2.5:10:50; 200:2.5:15:50;

[0158] The remaining steps and processes are all carried out with reference to Example 1 to obtain the compositions of this comparative example with different proportions of tanshinone ⅡA.

[0159] Referring to the above method, the effects of the compositions of different mass ratios of strychnine, atractylodes lactone III, tanshinone ⅡA, and ligusticum chuanxiong lactone A in this example on serum rheumatoid factor and inflammatory factors in rats with rheumatoid arthritis were compared. The results are shown in Table 5.

[0160] Table 5

[0161] Group RF (IU / ml) IL-17 (pg / ml) IFN-γ (pg / ml) TNF-α (pg / ml) IL-1β (pg / ml) IL-10 (pg / ml) Example 2 140.36±4.65 49.02±6.87 8.79±3.78 5.56±3.21 17.97±1.25 26.52±8.12 20：2.5：1：50 <![CDATA[179.37±4.09 * ]]> <![CDATA[86.34±6.54 * ]]> <![CDATA[37.48±5.01 * ]]> <![CDATA[9.97±8.71 * ]]> <![CDATA[29.35±3.09 * ]]> 19.82±3.98 20：2.5：2.5：50 148.30±5.09 60.03±4.87 9.23±3.09 6.32±5.01 19.44±3.45 24.34±5.01 20：2.5：5：50 141.92±3.83 51.96±9.08 8.81±1.90 5.53±5.67 17.72±5.06 26.01±7.23 20：2.5：10：50 151.63±7.65 54.26±7.01 13.01±3.21 7.19±2.09 19.54±7.01 23.06±4.57 20：2.5：15：50 <![CDATA[179.36±4.58 * ]]> <![CDATA[82.32±3.21 * ]]> <![CDATA[32.15±2.80 * ]]> <![CDATA[9.81±6.87 * ]]> <![CDATA[26.24±5.09 * ]]> <![CDATA[15.59±8.77 * ]]>

[0162] (Note: Compared with Example 2, * p<0.05)

[0163] It can be seen from Table 5 that when the ratio of tanshinone ⅡA to strychnine is 2.5~10:200, its effect in improving rheumatoid arthritis is equivalent to that of Example 2. When the ratio exceeds this range, its performance is significantly reduced compared with Example 2. This shows that within this range, there is a synergistic effect between the active compositions.

[0164] Example 5

[0165] This example is used to explore the effects of different ratios of brucine lactone A to strychnine, atractylodes lactone III, and tanshinone ⅡA on the performance of the composition in treating rheumatoid arthritis. Specifically, the content ratios of brucine to atractylodes lactone III, tanshinone ⅡA, and brucine lactone A in Example 1 are adjusted to be: 200:2.5:5:20; 200:2.5:5:30;

[0168] 200:2.5:5:50 (Example 1); 200:2.5:5:100; 200:2.5:5:120;

[0171] The remaining steps and processes are all referred to Example 1 to obtain compositions with different proportions of ligusticum chuanxiong lactone A in this comparative example.

[0172] Referring to the above method, the effects of the compositions of different mass ratios of strychnine: atractylodes lactone III, tanshinone ⅡA, and ligusticum chuanxiong lactone A in this example on serum rheumatoid factor and inflammatory factors in rats with rheumatoid arthritis were compared. The results are shown in Table 6.

[0173] Table 6

[0174] Group RF (IU / ml) IL-17 (pg / ml) IFN-γ (pg / ml) TNF-α (pg / ml) IL-1β (pg / ml) IL-10 (pg / ml) Example 2 140.36±4.65 49.02±6.87 8.79±3.78 5.56±3.21 17.97±1.25 26.52±8.12 20：2.5：5：20 <![CDATA[180.63±2.34 * ]]> <![CDATA[83.15±4.34 * ]]> <![CDATA[31.18±2.22 * ]]> <![CDATA[8.91±2.34 * ]]> <![CDATA[24.62±2.45 * ]]> <![CDATA[15.90±0.01 * ]]> 20：2.5：5：30 151.63±3.09 54.26±5.02 13.01±3.08 7.19±6.01 19.54±5.76 23.06±4.52 20：2.5：5：50 141.92±1.98 51.96±2.98 8.81±5.01 5.53±7.01 17.72±4.09 26.01±3.02 20：2.5：5：100 143.08±3.24 59.30±5.09 9.32±2.98 7.63±4.38 21.46±5.97 23.26±5.55 20：2.5：5：120 <![CDATA[169.76±6.05 * ]]> <![CDATA[83.39±7.09 * ]]> <![CDATA[35.67±7.09 * ]]> <![CDATA[10.07±7.01 * ]]> <![CDATA[31.01±3.21 * ]]> 18.22±8.18

[0175] (Note: Compared with Example 2, * p<0.05)

[0176] It can be seen from Table 6 that when the ratio of strychnine lactone A to strychnine is 30-100:200, its effect in improving rheumatoid arthritis is equivalent to that of Example 2. When the ratio exceeds this range, its performance is significantly reduced compared with Example 2. This shows that within this range, there is a synergistic effect between the active compositions.

[0177] Example 6

[0178] In order to further verify the active substances that are actually effective in the composition of the present application, the difference between this example and Example 1 is that the Atractylodes macrocephala extract is replaced with an equal amount of Atractylodes macrocephala lactone III or an equal amount of Atractylodes macrocephala lactone I, and the remaining steps and processes are referred to Example 1 to obtain the composition of this example, which are respectively recorded as S6-1 and S6-2.

[0179] The results of the efficacy of the comparative example 1 and the composition of this example are shown in Table 7.

[0180] Table 7

[0181] Group RF (IU / ml) IL-17 (pg / ml) IFN-γ (pg / ml) TNF-α (pg / ml) IL-1β (pg / ml) IL-10 (pg / ml) Example 1 141.92±4.42 51.96±6.65 8.81±0.56 5.53±1.21 17.72±2.18 26.01±2.65 S6-1 141.23±0.21 48.26±6.01 7.44±3.21 5.32±2.87 16.91±3.91 27.21±2.91 S6-2 <![CDATA[165.33±3.21 * ]]> <![CDATA[86.23±8.91 * ]]> <![CDATA[39.33±5.01 * ]]> <![CDATA[10.08±0.12 * ]]> <![CDATA[28.13±7.01 * ]]> 19.23±2.32

[0182] (Note: Compared with Example 1, * p<0.05)

[0183] As can be seen from Table 7, when the Atractylodes macrocephala extract is replaced with an equal amount of atractylodes lactone III, the effect of the composition in improving rheumatoid arthritis is not significantly different from that in Example 1, while when the Atractylodes macrocephala extract is replaced with an equal amount of atractylodes lactone I, the effect is significantly different from that in Example 1, which indicates that it is mainly atractylodes lactone III in the Atractylodes macrocephala extract that coordinates with other components to improve rheumatoid arthritis.

[0184] Example 7

[0185] In order to further verify the active substances that are actually effective in the composition of the present application, the difference between this embodiment and Example 1 is that the Salvia miltiorrhiza extract is replaced with an equal amount of Tanshinone ⅡA or an equal amount of Danshensu, and the remaining steps but processes are all referred to Example 1 to obtain the compositions of this embodiment, which are respectively recorded as S7-1 and S7-2.

[0186] The results of the efficacy of the comparative example 1 and the composition of this example are shown in Table 8.

[0187] Table 8

[0188] Group RF (IU / ml) IL-17 (pg / ml) IFN-γ (pg / ml) TNF-α (pg / ml) IL-1β (pg / ml) IL-10 (pg / ml) Example 1 141.92±4.42 51.96±6.65 8.81±0.56 5.53±1.21 17.72±2.18 26.01±2.65 S7-1 136.63±0.98 48.23±7.09 7.99±6.87 5.43±7.44 16.61±4.87 28.09±5.21 S7-2 <![CDATA[169.22±0.65 * ]]> <![CDATA[84.34±5.43 * ]]> <![CDATA[35.06±6.05 * ]]> <![CDATA[10.22±3.04 * ]]> <![CDATA[26.36±0.65 * ]]> 18.29±3.42

[0189] (Note: Compared with Example 1, * p<0.05)

[0190] As can be seen from Table 8, when the salvia miltiorrhiza extract is replaced by an equal amount of tanshinone ⅡA, the effect of the composition in improving rheumatoid arthritis is not significantly different from that in Example 1, while when the salvia miltiorrhiza extract is replaced by an equal amount of tanshinone, the effect is significantly different from that in Example 1, which indicates that tanshinone ⅡA and other components in the salvia miltiorrhiza extract synergistically improve rheumatoid arthritis.

[0191] Example 8

[0192] In order to further verify the active substances that are actually effective in the composition of the present application, the difference between this embodiment and Example 1 is that the Chuanxiong extract is replaced with an equal amount of ligusticum chuanxiong lactone A or an equal amount of ligusticum chuanxiong lactone I, and the remaining steps but processes are all based on Example 1 to obtain the compositions of this embodiment, which are respectively recorded as S8-1 and S8-2.

[0193] The results of the efficacy of the comparative example 1 and the composition of this example are shown in Table 9.

[0194] Table 9

[0195] Group RF (IU / ml) IL-17 (pg / ml) IFN-γ (pg / ml) TNF-α (pg / ml) IL-1β (pg / ml) IL-10 (pg / ml) Example 1 141.92±4.42 51.96±6.65 8.81±0.56 5.53±1.21 17.72±2.18 26.01±2.65 S8-1 140.01±2.09 46.55±5.02 7.38±5.01 5.31±5.87 17.58±3.09 27.07±2.45 S8-2 <![CDATA[179.05±5.02 * ]]> <![CDATA[85.43±2.87 * ]]> <![CDATA[27.01±4.34 * ]]> <![CDATA[9.78±3.21 * ]]> <![CDATA[26.54±1.98 * ]]> <![CDATA[19.12±7.01 * ]]>

[0196] As can be seen from Table 9, when the Chuanxiong extract is replaced with an equal amount of ligusticum chuanxiong lactone A, the effect of the composition in improving rheumatoid arthritis is not significantly different from that in Example 1, while when the Chuanxiong extract is replaced with an equal amount of ligusticum chuanxiong lactone I, there is a significant difference in the effect compared with Example 1, which indicates that ligusticum chuanxiong lactone A in the Chuanxiong extract is mainly synergistic with other components to improve rheumatoid arthritis.

[0197] In summary, the present invention finds that selecting strychnine (from Strychnos nux vomica or Strychnos nux vomica extract), atractylodes lactone III (from Atractylodes macrocephala or Atractylodes macrocephala extract), tanshinone ⅡA (from Salvia miltiorrhiza or Salvia miltiorrhiza extract) and ligusticum lactone A (from Chuanxiong or Chuanxiong extract) for compounding in a suitable proportion can achieve a good therapeutic effect on rheumatoid arthritis, which is equivalent to the effect of a complete traditional Chinese medicine composition consisting of Strychnos nux vomica, Atractylodes macrocephala, Salvia miltiorrhiza, Chuanxiong, Licorice, Codonopsis pilosula, Poria, Panax notoginseng, Achyranthes bidentata and Pheretima. This discovery reveals the key active ingredients in the traditional Chinese medicine composition for the treatment of rheumatoid arthritis, provides a scientific basis for the modernization and internationalization of traditional Chinese medicine, and a streamlined formula may be easier for patients to accept, thereby improving treatment compliance and satisfaction.

[0198] The present invention can achieve more accurate medication by precisely controlling the ratio of each effective ingredient, which not only ensures the therapeutic effect, but also reduces the side effects that may be caused by excessive ingredients, thereby improving the safety of the drug and the tolerance of patients.

[0199] It should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention rather than to limit it. 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 composition for treating rheumatoid arthritis, characterized in that: It is composed of Chinese medicinal materials or their extracts containing the following active ingredients: Strychnine-containing seeds of Strychnos nux vomica or their extracts, atractylodes macrocephala or their extracts containing atractylodes lactone III, salvia miltiorrhiza or their extracts containing tanshinone ⅡA, chuanxiong or their extracts containing chuanxiong lactone A; Among them, the content ratio of strychnine to atractylodes lactone III, tanshinone ⅡA, and ligusticum chuanxiong lactone A is 200:1~5:2.5~10:30~100.

2. The composition for treating rheumatoid arthritis according to claim 1, characterized in that: The content ratio of strychnine to atractylodes lactone III is 200:1-2.

5.

3. The composition for treating rheumatoid arthritis according to claim 2, characterized in that: The content ratio of strychnine to tanshinone ⅡA is 200:2.5~5.

4. The composition for treating rheumatoid arthritis according to claim 3, characterized in that: The content ratio of strychnine to ligusticum lactone A is 200:30-50.

5. A composition for treating rheumatoid arthritis, characterized in that: It is composed of the following components: Strychnine-containing Strychnine or Strychnine extract, Atractylodes macrocephala or Atractylodes macrocephala extract containing Atractylodes lactone III, Salvia miltiorrhiza or Salvia miltiorrhiza extract containing Tanshinone ⅡA, Chuanxiong or Chuanxiong extract containing Chuanxiong lactone A, Licorice, Codonopsis pilosula, Poria, Panax notoginseng, Achyranthes bidentata, Pheretima; The content ratio of strychnine to atractylodes lactone III, tanshinone ⅡA, and ligusticum chuanxiong lactone A is 200:1~5:2.5~10:30~100; The mass ratio of strychnine in the strychnine or strychnine extract to liquorice, codonopsis, tuckahoe, panax notoginseng, achyranthes bidentata and earthworm is 0.1:5~20:10~30:10~20:5~20:5~15:0.5~5.

6. The composition for treating rheumatoid arthritis according to claim 5, characterized in that: The mass ratio of strychnine in the strychnine or strychnine extract to liquorice, codonopsis pilosula and poria is 0.1:5-15:10-15:10-15.

7. The composition for treating rheumatoid arthritis according to claim 6, characterized in that: The mass ratio of brucine in the strychnine or strychnine extract to notoginseng and Achyranthes bidentata is 0.1:10-20:5-10.

8. The composition for treating rheumatoid arthritis according to claim 7, characterized in that: The mass ratio of brucine to earthworm in the strychnine or strychnine extract is 0.1:1-5.

9. Use of the composition according to any one of claims 1 to 4 or 5 to 8 in the preparation of a drug for treating rheumatoid arthritis.

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