Composition for treating rheumatoid arthritis

Abstract

The invention provides a composition for treating arthritis diseases. The composition comprises luteolin and scoparone as active ingredients. The mass ratio of the luteolin to the scoparone in the composition is 1: (3.6-21.6). Experiments show that the composition disclosed by the invention has a good anti-arthritis effect and has no obvious toxicity. The composition can be used for preparing effective and safe anti-arthritis drugs.

Description

Technical Field

[0001] This invention relates to the field of pharmaceutical technology, and more specifically to a composition for treating arthritis, particularly rheumatoid arthritis. Background Technology

[0002] Arthritis is a disease that indicates damage or inflammation of one or more joints. It often manifests as pain, swelling, heat, redness, and limited mobility. Many different forms of arthritis exist, with rheumatoid arthritis (RA) being one of the most common. RA is an autoimmune disease with a long course, unclear cause, and a high rate of disability. Typical clinical manifestations of RA include symmetrical polyarthritis, often accompanied by pain, fatigue, joint dysfunction, and mood disorders. It can lead to disability, deformity, and various complications, severely impacting patients' health and quality of life. Therefore, RA is sometimes referred to as "the cancer that doesn't kill." In recent years, with economic development, improved living standards, and an aging population, the number of RA patients has increased rapidly worldwide, becoming a global disease. Currently, the global prevalence of RA is approximately 1.0%, with an annual incidence rate of 0.25%–0.5%. In 2010, RA caused approximately 49,000 deaths globally. my country has approximately 5 million RA patients, with an average age of onset of only 45 years. RA is one of the leading causes of disability and loss of labor capacity in my country. Therefore, developing effective drugs for the treatment of RA has significant social implications.

[0003] Currently, medications for treating rheumatoid arthritis (RA) mainly include disease-modifying antirheumatic drugs (DMARDs, such as methotrexate and leflunomide), bone-targeting drugs (denosumab), nonsteroidal anti-inflammatory drugs (NSAIDs, such as aspirin, indomethacin, and ibuprofen), and glucocorticoids (such as triamcinolone, dexamethasone palmitate, and prednisolone). However, these drugs have many side effects. For example, long-term use of NSAIDs can cause gastrointestinal side effects and increase the incidence of cardiovascular disease; a small number of patients may experience kidney damage, bone marrow suppression, and mental disorders. DMARDs have a certain inhibitory effect on the human immune system, and taking these drugs can cause serious side effects such as infections. In addition, DMARDs have also been found to have side effects such as gastrointestinal damage, hepatotoxicity, and pulmonary toxicity. Long-term use of glucocorticoids can lead to Cushing's syndrome, hypertension, and slowed wound healing. This not only endangers the patient's life and health but also places a heavy burden on the patient's family.

[0004] The Tang Dynasty medical text *Qianjin Fang* records a formula composed of dried fruit of *Rosa multiflora* and dried flower buds or unopened flowers of *Lonicera japonica*, used to treat inflammation-related diseases by soaking them in alcohol. Studies have shown that the ethanol extract of this formula has significant anti-RA activity in rat models without significant toxicity, suggesting its potential to be developed into a highly effective, low-toxicity drug for treating RA. However, due to the complexity of the components of traditional Chinese medicine compound formulas and the unclear identification of their active ingredients, they are not easily developed into modern formulations and promoted for international application.

[0005] Therefore, there is an urgent need to provide a drug with a clear chemical composition and controllable quality for the treatment of arthritis, especially rheumatoid arthritis (RA). Summary of the Invention

[0006] To address the shortcomings of existing medications for treating arthritis, particularly rheumatoid arthritis (RA), this application provides a formulation of effective components from traditional Chinese medicine, wherein the effective components consist of luteolin and artemisinin. Compared to traditional Chinese medicine compound formulas, this formulation of effective components has a clear chemical composition and controllable quality; compared to single compounds, it retains the characteristics of traditional Chinese medicine—multiple components and multiple targets—and has advantages in addressing arthritis, particularly RA, with its complex etiologies.

[0007] Luteolin is a flavonoid compound with the molecular formula C1. 15 H 10 O6, with a molecular weight of 286.24, has the following structural formula: Figure 1 As shown, luteolin possesses various pharmacological activities, such as anti-inflammatory, anticancer, antioxidant, immunomodulatory, angiogenic, and neuroprotective effects. Animal experiments have shown that luteolin has anti-RA effects in both rat and mouse models.

[0008] Artemisia lactone is a coumarin compound with the molecular formula C. 11 H 10 O4, with a molecular weight of 206.19, has the following structural formula: Figure 2 As shown in the figure. Studies have shown that artemisinin has pharmacological effects such as anti-inflammatory, antioxidant, and lipid-lowering effects.

[0009] The inventors of this application have discovered that the combined use of luteolin and artemisinin exhibits significantly better anti-arthritis effects compared to the use of either compound alone. The two compounds have a synergistic effect and low toxicity.

[0010] On one hand, the present invention provides a composition for treating arthritis, comprising luteolin and artemisinin as active ingredients.

[0011] The term "arthritis" as used in this article refers to autoimmune diseases and / or inflammatory diseases manifesting as joint inflammation.

[0012] In one embodiment, the arthritis is rheumatoid arthritis.

[0013] In one implementation, arthritis affects one or more joints in the hand, knee, hip, spine, wrist, ankle, hip, toes, and / or elbow.

[0014] In one embodiment, the mass ratio of luteolin to artemisinin in the composition is luteolin:artemisinin = 1:2 to 30, preferably 1:3 to 25. In a more preferred embodiment, the mass ratio of luteolin to artemisinin in the composition is luteolin:artemisinin = 1:3.6 to 21.6. In an even more preferred embodiment, the mass ratio of luteolin to artemisinin in the composition is luteolin:artemisinin = 1:6.

[0015] In one embodiment, the active ingredients in the composition consist of luteolin and artemisinin. In another embodiment, the active ingredients in the composition may include, in addition to luteolin and artemisinin, other active agents used to treat arthritis, particularly rheumatoid arthritis. These other active agents used to treat arthritis, particularly rheumatoid arthritis, are known to those skilled in the art and do not include traditional Chinese medicines, medicinal herbs, or extracts thereof containing luteolin and / or artemisinin.

[0016] One of the objectives of this invention is to provide a composition with clearly defined active ingredients and controllable quality. Therefore, the active ingredients in the composition of this invention are clearly defined and specific, and do not include traditional Chinese medicines, medicinal materials or their extracts containing luteolin and / or artemisinin.

[0017] In one implementation, luteolin and artemisinin can exist individually or in combination.

[0018] On the other hand, the present invention provides a pharmaceutical preparation comprising the above-described composition for treating arthritis and pharmaceutically acceptable excipients.

[0019] In one embodiment, the dosage form of the pharmaceutical preparation includes, but is not limited to, tablets, capsules, granules, powders, suspensions, solutions, injections, ointments, creams, gels, tinctures, and liniments. All of the above dosage forms can be prepared according to conventional methods in the pharmaceutical field.

[0020] In one embodiment, the pharmaceutical preparation can be administered orally, by injection (intravenous, intramuscular, or subcutaneous) or by topical application.

[0021] In one embodiment, in the pharmaceutical preparation, luteolin and artemisinin may be formulated into the same or different dosage forms, the dosage forms being selected from tablets, capsules, granules, powders, suspensions, solutions, injections, ointments, creams, gels, tinctures, and liniments.

[0022] In one embodiment, in the pharmaceutical formulation, luteolin and artemisinin can be formulated into dosage forms for administration via the same or different routes, selected from oral, injectable, or topical administration.

[0023] In one embodiment, in the pharmaceutical formulation, luteolin and artemisinin are present in the same or different dosage forms for oral administration, such as tablets, capsules, granules, powders, or oral solutions.

[0024] In one embodiment, luteolin and artemisinin are present in the pharmaceutical formulation in dosage forms intended for injection (e.g., intravenous, intramuscular, or subcutaneous injection).

[0025] In one embodiment, luteolin and artemisinin are present in the pharmaceutical formulation in topical dosage forms, such as ointments, creams, gels, tinctures, lotions, etc.

[0026] In one embodiment, in the pharmaceutical formulation, luteolin is present in a dosage form for oral administration; while artemisinin is present in a dosage form for injection or topical administration.

[0027] In one embodiment, artemisinin is present in the pharmaceutical formulation in an oral administration form; while luteolin is present in an injection or topical administration form.

[0028] On the other hand, the present invention provides the use of combinations of luteolin and artemisinin, such as the above-described composition containing luteolin and artemisinin as active ingredients, in the preparation of a medicament for treating arthritis.

[0029] In one embodiment, the arthritis is rheumatoid arthritis.

[0030] In one embodiment, luteolin and artemisinin are combined at a mass ratio of luteolin:artemisinin = 1:2 to 30. In a preferred embodiment, luteolin and artemisinin are combined at a mass ratio of luteolin:artemisinin = 1:3 to 25. In a more preferred embodiment, luteolin and artemisinin are combined at a mass ratio of luteolin:artemisinin = 1:3.6 to 21.6. In an even more preferred embodiment, luteolin and artemisinin are combined at a mass ratio of luteolin:artemisinin = 1:6.

[0031] In one embodiment, the combination or composition of luteolin and artemisinin can be formulated into various pharmaceutical dosage forms, including but not limited to tablets, capsules, granules, powders, solutions, injections, ointments, creams, gels, tinctures, and liniments. All of the above dosage forms can be prepared according to conventional methods in the pharmaceutical field.

[0032] In one embodiment, the combination or composition of luteolin and artemisinin can be formulated into a medicament for administration via an appropriate route, including but not limited to oral, injectable, and topical forms.

[0033] In one embodiment, luteolin and artemisinin, alone or together, are formulated into a drug for oral administration, for example in the form of tablets, capsules, granules, powders or oral solutions.

[0034] In one embodiment, luteolin and artemisinin, alone or together, are formulated into a drug for injection, such as intravenous, intramuscular or subcutaneous injection.

[0035] In one embodiment, luteolin and artemisinin are formulated, alone or together, into topical dosage forms, such as ointments, creams, gels, tinctures, lotions, etc.

[0036] In one embodiment, luteolin is formulated as a drug for oral administration; while artemisinin is formulated as a drug for injection or topical administration.

[0037] In one embodiment, artemisinin is formulated as a drug for oral administration; while luteolin is formulated as a drug for injection or topical administration.

[0038] In one embodiment, the combination or composition of luteolin and artemisinin can be used in combination with other drugs for treating arthritis. In one embodiment, the combination or composition of luteolin and artemisinin can be used in combination with other antirheumatoid arthritis drugs for treating rheumatoid arthritis. The other drugs for treating arthritis and the other antirheumatoid arthritis drugs are known to those skilled in the art, but do not include traditional Chinese medicines, medicinal herbs, or extracts thereof containing luteolin and / or artemisinin.

[0039] The technical solution of the present invention has the following beneficial effects:

[0040] 1. The present invention provides a novel composition for treating arthritis, particularly rheumatoid arthritis.

[0041] 2. The composition of the present invention has a significant inhibitory effect on the proliferation of synovial cells in patients with arthritis, and the effect is significantly stronger than that of luteolin and artemisinin used alone. Calculations showed that luteolin and artemisinin have a synergistic effect in this cell model.

[0042] 3. The composition of the present invention can effectively improve arthritis symptoms in CIA mice, and its anti-arthritis effect is significantly better than that of the positive control drug leflunomide.

[0043] 4. The composition of the present invention can significantly alleviate the weight loss in CIA mice, indicating that it has low toxicity and is significantly less toxic than the positive control drug leflunomide.

[0044] 5. This invention provides a novel application of the combination of luteolin and artemisinin in the preparation of medicaments for treating arthritis, particularly rheumatoid arthritis. Attached Figure Description

[0045] Figure 1 The structural formula of luteolin.

[0046] Figure 2 The structural formula of artemisinin.

[0047] Figure 3 Effects of luteolin, artemisinin, and their combination (Lu+Sco) on RA-FLS cell survival. Compared with the IL-6 / sIL-6R stimulation group, *P<0.05, **P<0.01. &q>1.15.

[0048] Figure 4 Photographs of the hind feet of mice after administration of Lu+Sco and the positive control drug leflunomide.

[0049] Figure 5 X-ray images of the hind paws of mice after administration of Lu+Sco and the positive control drug leflunomide.

[0050] Figure 6 Effects of Lu+Sco and the positive control drug leflunomide on hindfoot swelling in arthritic mice. Compared with the normal group, ##P<0.01. Compared with the model group, **P<0.01. Compared with the positive control group, &P<0.05; &&P<0.01.

[0051] Figure 7 Effects of Lu+Sco and the positive control drug leflunomide on arthritis scores in arthritic mice. Compared with the normal group, ##P<0.01. Compared with the model group, **P<0.01. Compared with the positive control group, &&P<0.01.

[0052] Figure 8Effects of Lu+Sco and the positive control drug leflunomide on serum inflammatory factor levels in arthritic mice. Compared with the normal group, ##P<0.01. Compared with the model group, **P<0.01. Compared with the positive control group, &P<0.05; &&P<0.01.

[0053] Figure 9 Effects of Lu+Sco and the positive control drug leflunomide on body weight in arthritic mice. Compared with the normal group, ##P<0.01. Compared with the model group, **P<0.01. Compared with the positive control group, &&P<0.01. Detailed Implementation

[0054] The present invention will be further described below with reference to specific embodiments, so that those skilled in the art can better understand the present invention. However, it should not be construed as the scope of the above-mentioned subject matter of the present invention being limited to the following examples. All technical solutions implemented based on the above content of the present invention fall within the scope of the present invention.

[0055] Example

[0056] Example 1: Effects of luteolin, artemisinin and their combinations on synovial cell survival in patients with arthritis.

[0057] 1.1 Experimental Materials

[0058] Medicines and reagents

[0059] (1) Luteolin (Lu): Luteolin ≥98%, produced by Chengdu Efa Biotechnology Co., Ltd.

[0060] (2) Scoparone (Sco): Scoparone ≥98%, produced by Chengdu Efa Biotechnology Co., Ltd.

[0061] (3) Synoviocyte Medium, produced by ScienCell Research Laboratories, USA.

[0062] (4) 0.5% (mass percentage) trypsin-EDTA (10X): contains 0.5% (mass percentage) EDTA, 100mL / bottle, manufactured by Gibco, USA.

[0063] (5) Fetal bovine serum: 500mL / bottle, produced by Gibco, USA.

[0064] (6) Interleukin-6 (IL-6) and soluble interleukin-6 receptor (sIL-6R), manufactured by PeproTech, USA.

[0065] (7) Cell Counting Kit-8 (CCK-8) reagent kit, manufactured by Dojindo Corporation, Japan.

[0066] (8) Leflunomide: Leflunomide ≥98%, produced by Shanghai Maclean Biochemical Technology Co., Ltd.

[0067] cell lines

[0068] The cell line used in this embodiment is synovial cells from arthritis patients (RA-FLS), which was purchased from Cell Applications, Inc., USA.

[0069] 1.2 Cellular Experimental Methods

[0070] In this experiment, the CCK-8 assay kit was used to detect the effects of luteolin, artemisinin and their combinations on the survival rate of synovial cells (RA-FLS) in patients with arthritis.

[0071] RA-FLS cells were cultured in Synoviocyte Medium containing 5% (w / v) FBS and 100 U / mL penicillin-streptomycin at 37°C in a 5% (v / v) CO2 incubator. Once the cells reached the logarithmic growth phase, they were digested with 0.25% trypsin to prepare a single-cell suspension, which was then seeded into 96-well plates at a concentration of 3000 cells per well.

[0072] After 12 hours of cell culture, cells were divided into two groups: a control group (IL-6 / sIL-6R stimulation group) and treatment groups (luteolin (Lu), four artemisinin (Sco), four luteolin + artemisinin (Lu+Sco), and a positive control group (Lef10). RA-FLS cells were first treated for 1 hour with 0.1% DMSO (IL-6 / sIL-6R stimulation group), 10 μM luteolin (Lu), 50 / 100 / 200 / 300 μM artemisinin (Sco), 10 μM luteolin + 50 / 100 / 200 / 300 μM artemisinin (Lu+Sco), or 10 μM leflunomide (Lef10, positive control group). Subsequently, each group was stimulated with 100 ng / ml IL-6 / sIL-6R for 24 hours / 48 hours.

[0073] After 24 / 48 hours, add 10 μL of CCK8 assay reagent to each well and continue culturing for 2 hours. Measure the absorbance at OD 450 nm using a full-wavelength microplate reader (Thermo Fisher Scientific), and then calculate the RA-FLS cell viability using the following formula:

[0074] RA-FLS cell viability = T / C * 100%, where C is the absorbance value of the control group and T is the absorbance value of the drug-treated group.

[0075] Methods for calculating synergistic effects:

[0076] Cell growth inhibition rate = 1 - cell survival rate in the treatment group

[0077] The cell growth inhibition rate was calculated using the Jin formula (Jin Zhengjun. Additive effects in combined drug use. Chinese Journal of Pharmacology. 1980; 1:70-76), where q = E(A+B) / [EA+(1-EA)×EB], E(A+B) is the cell growth inhibition rate when the two drugs are used together, and EA and EB are the cell growth inhibition rates when each drug is used alone. When q < 0.85, it indicates that the two drugs are antagonistic; when q > 1.15, it indicates that the two drugs have a synergistic effect; and when q = 0.85 to 1.15, it indicates that the effects of the two drugs are additive.

[0078] 1.3 Results of Cell Experiments

[0079] according to Figure 3 It was observed that luteolin or artemisinin alone reduced the survival rate of RA-FLS cells stimulated by IL-6 / sIL-6R, and the survival rate of RA-FLS cells decreased significantly after treatment with luteolin + artemisinin. These results indicate that the combined use of luteolin and artemisinin significantly inhibits the growth of RA-FLS cells, and the combined effect is significantly stronger than that of luteolin or artemisinin alone.

[0080] Computational synergistic effects:

[0081] In this experiment, the composition of the present invention consisted of luteolin (10 μM) and artemisinin (50 / 100 / 200 / 300 μM), with a molar ratio of luteolin:artemisinin = 1:5–30. The molar mass of luteolin was 286.24 g / mol; the molar mass of artemisinin was 206.19 g / mol. After conversion, the mass ratio of the two was luteolin:artemisinin = 1:3.6–21.6. Calculations showed that at this mass ratio, the combined use of luteolin and artemisinin could inhibit the proliferation of RA-FLS cells, and the q-value of all combinations was greater than 1.15, indicating that the combined use of luteolin and artemisinin had a synergistic effect.

[0082] Example 2: Determination of the anti-arthritis effect of the combination of luteolin and artemisinin on a mouse model.

[0083] 2.1 Animal Experiment Materials

[0084] Medicines and reagents

[0085] (1) Luteolin (Lu): Luteolin ≥98%, produced by Chengdu Efa Biotechnology Co., Ltd.

[0086] (2) Scoparone (Sco): Scoparone ≥98%, produced by Chengdu Efa Biotechnology Co., Ltd.

[0087] (3) Leflunomide: Leflunomide ≥98%, produced by Shanghai Maclean Biochemical Technology Co., Ltd.

[0088] (4) Complete Freund's adjuvant (CFA): manufactured by Chondrex, Inc., USA.

[0089] (5) Bovine type II collagen (CII): Produced by Chondrex, USA.

[0090] (6) Interleukin-6 (IL-6), interleukin-1β (IL-1β), and tumor necrosis factor-α (TNF-α) enzyme-linked immunosorbent assay (ELISA) kit: manufactured by Hangzhou Lianke Biotechnology Co., Ltd.

[0091] laboratory animals

[0092] DBA-1J male mice, 7 weeks old, were purchased from The Jackson Laboratory in the United States. They were housed in a sterile environment with constant temperature and pressure at the Laboratory Animal Centre of Hong Kong Baptist University.

[0093] 2.2 Animal Experiment Methods

[0094] 2.2.1 Establishment of a collagen-induced mouse model of arthritis (CIA)

[0095] DBA / 1J mice were randomly divided into four groups: normal group, model group, Lu+Sco group, and positive control group. Except for the normal group, the mice in the other groups were immunized by subcutaneous injection of 200 μg of bovine type II collagen (CII) emulsified with complete Freund's adjuvant (CFA) at the tail on day 0.

[0096] 2.2.2 Animal feeding and drug administration

[0097] Mice were housed in a standard environment (25±2℃, humidity: 60±10%, 12h diurnal cycle). All drugs were prepared using 0.5% CMC-Na. From day 18 to day 46 of the experiment, the three groups of model mice were administered 0.5% CMC-Na by gavage (model group); luteolin 5 mg / kg / day and artemisinin 30 mg / kg / day (Lu+Sco group); and leflunomide 4 mg / kg / day (human clinical equivalent dose, positive control group). Simultaneously, the normal control group was administered 0.5% CMC-Na by gavage. Administered once daily for 28 days.

[0098] 2.2.3 Recording of mouse body weight

[0099] Starting from day 0 of the experiment, the mice's weight changes were measured and recorded weekly.

[0100] 2.2.4 Evaluation of joint swelling and pathological changes

[0101] Starting from the first day of drug administration (day 18 of the experiment), the diameter of the bilateral hind paw ankle joints was measured every 4 days using calipers to evaluate the degree of swelling in the hind paw ankle joints of the mice, and to determine the gross severity of the disease and the treatment effect. On day 46 of the experiment, the hind paws of the mice in each group were photographed and the degree of hind paw swelling in each group was compared. Subsequently, an X-ray system ( X-ray images of the hind paws of mice were taken using an MX-20 (UK) to observe the pathological changes in the ankle joints of the hind paws of mice in each group.

[0102] 2.2.5 Evaluation of lesion severity

[0103] Starting from day 30 of the experiment, every 4 days, an arthritis score (0-4 points) was calculated based on the erythema and swelling of the mice's limbs. Scoring criteria: 0 = Normal, no visible arthritis symptoms; 1 = Mild, visible swelling or patchy erythema in the ankle / toe, regardless of the number of patches; 2 = Moderate, moderate swelling in the toe joints, toes, or ankles; 3 = Severe swelling of the ankle joint or swelling below the ankle; 4 = Swelling of the entire paw or severe joint deformity. The sum of the scores for all four limbs was the arthritis score for each mouse, with a maximum total score of 16 points per mouse.

[0104] 2.2.6 Detection of IL-6, IL-1β and TNF-α in serum

[0105] On day 46, mice were anesthetized with isoflurane (induction dose: oxygen flow rate, 1 L / min; isoflurane, 3%; maintenance dose: oxygen flow rate, 0.3 L / min; isoflurane, 1%). Blood was collected and serum was obtained. The levels of IL-6, IL-1β and TNF-α in mouse serum were determined according to the ELISA kit instructions.

[0106] 2.3 Experimental Results

[0107] 2.3.1 Effects of the combination of luteolin and artemisinin on arthritis in CIA mice

[0108] according to Figure 4 It was evident that, compared to the normal group of mice, the model group mice exhibited significant hind paw swelling. Both luteolin + artemisinin (Lu + Sco group) and the clinically equivalent dose of leflunomide (positive control group) significantly reduced the degree of paw swelling in CIA mice. According to... Figure 5X-ray results showed that the model group mice exhibited bone erosion, joint damage, tissue edema, and osteophyte formation in the hindfoot ankle joint; luteolin + artemisinin (Lu + Sco group) and leflunomide (positive control group) significantly alleviated these pathological phenomena. Figure 6 The results of joint swelling showed that the model group mice had the highest degree of ankle joint swelling; luteolin + artemisinin (Lu + Sco group) significantly reduced ankle joint swelling in CIA mice, and the effect was significantly better than leflunomide (positive control group). Figure 7 Joint lesion scores showed that the model group had the highest degree of lesion; luteolin + artemisinin (Lu + Sco group) significantly reduced the degree of lesion in CIA mice, and the effect was significantly better than leflunomide (positive control group).

[0109] 2.3.2 Effects of the combination of luteolin and artemisinin on the serum levels of IL-6, IL-1β, and TNF-α in CIA mice

[0110] according to Figure 8 It was observed that the levels of inflammatory factors IL-6, IL-1β, and TNF-α in the serum of model group mice were significantly higher than those in the normal group. Both luteolin + artemisinin (Lu + Sco group) and the clinically equivalent dose of leflunomide (positive control group) significantly downregulated the levels of these three inflammatory factors in the serum of arthritic mice. Luteolin + artemisinin showed significantly better inhibitory effects on IL-6 and TNF-α than leflunomide, while its inhibitory effect on IL-1β was comparable to that of leflunomide.

[0111] 2.3.3 Effects of the combination of luteolin and artemisinin on body weight in CIA mice

[0112] After modeling, as the mice's arthritis worsened, their food intake decreased, and their weight also decreased accordingly. According to... Figure 9 As can be seen, the body weight of the model group mice was significantly lower than that of the normal group mice, indicating that CIA modeling leads to weight loss in mice. Simultaneously, the potential toxicity of the drug may also contribute to the weight loss in mice. According to... Figures 4 to 8 The results showed that luteolin + artemisinin (Lu + Sco group) and the clinically equivalent dose of leflunomide (positive control group) both significantly alleviated arthritis symptoms in mice. Figure 9As can be seen, the body weight of mice in the Lu+Sco group was significantly higher than that of mice in the model group and close to that of mice in the normal group, while there was no significant difference in body weight between mice in the positive control group and mice in the model group. These results indicate that the combination of luteolin and artemisinin can not only improve arthritis symptoms in mice but also restore the body weight of CIA mice to near-normal levels. Leflunomide, on the other hand, failed to improve the weight loss in CIA mice, suggesting that the combination of luteolin and artemisinin did not exhibit significant toxic side effects, while leflunomide is toxic and affects the body weight of mice. Therefore, the combination described in this invention is less toxic than leflunomide.

[0113] Clearly, the combination or composition of the present invention has a significant inhibitory effect on the proliferation of synovial cells in patients with arthritis, and the effect is significantly stronger than that of luteolin and artemisinin alone; calculations showed that luteolin and artemisinin have a synergistic effect in this cell model. The combination or composition of the present invention can effectively improve arthritis symptoms in CIA mice and significantly alleviate the weight loss in CIA mice, indicating that the combination or composition of the present invention has low toxicity. In addition, the combination or composition of the present invention has better anti-arthritis efficacy and lower toxicity compared with the positive control drug leflunomide. The above experimental results indicate that the combination or composition of luteolin and artemisinin can be formulated into an effective and low-toxic drug for treating arthritis, especially anti-rheumatoid arthritis.

Claims

1. A composition for treating arthritis, particularly rheumatoid arthritis, comprising luteolin and artemisinin as active ingredients.

2. The composition according to claim 1, characterized in that, The mass ratio of luteolin to artemisinin in the composition is 1:2 to 30, preferably 1:3.6 to 21.6, and more preferably 1:

6.

3. A pharmaceutical preparation comprising the composition according to claim 1 or 2, and pharmaceutically acceptable excipients.

4. The pharmaceutical preparation according to claim 3, characterized in that, The dosage form of the pharmaceutical preparation is selected from tablets, capsules, granules, powders, suspensions, solutions, injections, ointments, creams, gels, tinctures, and liniments.

5. The pharmaceutical preparation according to claim 3, which may be formulated for oral, injectable or topical administration.

6. Use of the combination of luteolin and artemisinin in the preparation of a medicament for the treatment of arthritis, particularly rheumatoid arthritis.

7. The use according to claim 6, characterized in that, The mass ratio of luteolin to artemisinin in the composition is 1:2 to 30, preferably 1:3.6 to 21.6, and more preferably 1:

6.

8. The use according to claim 6, characterized in that, The composition is formulated into a pharmaceutical dosage form selected from tablets, capsules, granules, suspensions, oral liquids, injections, ointments, creams, gels, tinctures, and liniments.

9. The use according to claims 6 to 8, characterized in that, The composition is formulated as a medicine for oral, injectable or topical administration.

10. The use according to claims 6 to 9, characterized in that, The drug is used in combination with other anti-arthritis drugs to treat arthritis.

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