Application of benzophenone compounds in preparation of anti-rheumatoid arthritis drugs

Abstract

The invention discloses an application of a halophenol compound 2, 4 ', 5'-trihydroxy-5, 2 '-dibromo benzophenone as a medicament for resisting rheumatoid arthritis. The compound can effectively relieve the ankle joint swelling degree of rheumatoid arthritis rats, relieve cartilage injury and reduce the arthritis score, main treatment indexes such as MMP-9, IL-17A, IL-1beta and IgM are all superior to those of a clinical medicine celecoxib, the reduction of the spleen coefficient is remarkably superior to that of celecoxib, and the important application prospect in the aspect of treating rheumatoid arthritis is shown.

Description

Technical Field

[0001] This invention belongs to the field of pharmaceutical applications of compounds, and relates to a benzophenone halogenated phenolic compound with the effect of preventing and treating rheumatoid arthritis, particularly the application of compound 2,4',5'-trihydroxy-5,2'-dibromobenzophenone (LF1) in the preparation of anti-rheumatoid arthritis drugs. Background Technology

[0002] Rheumatoid arthritis (RA) is a chronic, progressive inflammatory autoimmune disease. Early symptoms include swelling and pain in the affected joints, which then progresses to cartilage destruction and bone erosion, ultimately leading to joint deformities and impaired function. As RA progresses, it can affect other organs, causing serious complications.

[0003] Due to its high incidence, high disability rate, and poor cure rate, rheumatoid arthritis (RA) is often referred to as "the cancer that doesn't kill." Insufficient early prevention and treatment can lead to limited limb movement and even loss of earning capacity, placing a heavy burden on families and society as a whole. Therefore, strengthening research on the diagnosis, treatment, and pathogenesis of RA, and further developing new drugs with specific therapeutic effects, is urgently needed to improve the treatment outcomes of this disease.

[0004] Currently, the main treatment strategy for rheumatoid arthritis (RA) is to control inflammation and slow joint destruction, which can only relieve symptoms but cannot cure the disease. Commonly used drugs in clinical practice include glucocorticoids and nonsteroidal anti-inflammatory drugs (NSAIDs), such as methotrexate, sulfasalazine, leflunomide, hydroxychloroquine, olanofen, and celecoxib. Long-term use may lead to serious adverse reactions such as gastrointestinal irritation (e.g., nausea, vomiting, mucositis, and diarrhea), bone marrow suppression (e.g., anemia and neutropenia), liver and kidney damage (e.g., hepatitis, hematuria, proteinuria), pulmonary fibrosis, and hair loss. Therefore, there are currently no particularly effective and specific drugs for this disease in clinical practice. Given the dangers of RA, finding suitable treatment drugs has become a hot research topic in the medical field.

[0005] The pathogenesis of rheumatoid arthritis (RA) mainly includes immune dysregulation, bone erosion, and macrophage imbalance. The halogenated phenol compound 2,4',5'-trihydroxy-5,2'-dibromobenzophenone possesses strong anti-inflammatory, immunomodulatory, and antioxidant pharmacological effects. The diversity of its activities aligns well with the pathogenesis of RA, demonstrating significant potential for its development and application.

[0006] However, to date, there are no reports, either domestically or internationally, on the anti-rheumatoid arthritis pharmacological effects of compound 2,4',5'-trihydroxy-5,2'-dibromobenzophenone. Summary of the Invention

[0007] The object of this invention is to provide a new pharmaceutical use for the compound 2,4',5'-trihydroxy-5,2'-dibromobenzophenone.

[0008] Specifically, the present invention provides the use of the halogenated phenol compound 2,4',5'-trihydroxy-5,2'-dibromobenzophenone in the preparation of an antirheumatoid arthritis drug.

[0009] This invention is the first discovery of the anti-rheumatoid arthritis pharmacological activity of compound 2,4',5'-trihydroxy-5,2'-dibromobenzophenone, which has important development and application value.

[0010] The compound 2,4',5'-trihydroxy-5,2'-dibromobenzophenone is abbreviated as LF1, and its structural formula is as follows:

[0011] The compound 2,4',5'-trihydroxy-5,2'-dibromobenzophenone is a polyphenol compound well known to those skilled in the art, and its preparation method is also well known and easily implemented by those skilled in the art.

[0012] Providing a novel antirheumatoid arthritis drug is another objective of this invention.

[0013] This invention provides a novel antirheumatoid arthritis drug containing an effective dose of the compound 2,4',5'-trihydroxy-5,2'-dibromobenzophenone as the active pharmaceutical ingredient.

[0014] The novel antirheumatoid arthritis drug of the present invention, which uses compound 2,4',5'-trihydroxy-5,2'-dibromobenzophenone as the active ingredient, further includes a common pharmaceutical carrier or excipient used in combination with the active ingredient.

[0015] Furthermore, the drug can be formulated into various conventional pharmaceutical preparations such as tablets, injections, pellets, solid dispersions, or capsules, depending on different needs.

[0016] This invention establishes a rat model of rheumatoid arthritis (CIA) induced by fully Freund's adjuvant emulsified bovine type II collagen. Different doses of the compound tablets were administered by gavage, and various indicators such as rat body weight, ankle swelling, bone damage, and inflammatory factors were monitored during the process. The results confirmed the protective effect of the compound on rats with rheumatoid arthritis, and the experimental results show that the compound has important application prospects in the prevention and treatment of rheumatoid arthritis.

[0017] The main innovation of this invention is as follows:

[0018] 1) Compound 2,4',5'-trihydroxy-5,2'-dibromobenzophenone has strong anti-inflammatory, immunomodulatory, and antioxidant effects, and its diverse activities are highly consistent with the pathogenesis of rheumatoid arthritis (RA).

[0019] 2) Compound 2,4',5'-trihydroxy-5,2'-dibromobenzophenone can effectively reduce ankle swelling in rats with rheumatoid arthritis, alleviate cartilage damage and bone erosion, and reduce arthritis scores. Its main therapeutic indicators, such as MMP-9, IL-17A, IL-1β, and IgM, are superior to those of celecoxib, a current first-line clinical drug. Attached Figure Description

[0020] Figure 1 This is X-ray monitoring of changes in the ankle joint of rats.

[0021] Figure 2 The results were obtained by comparing the pathological findings of the ankle joint synovium in each group of rats under electron microscopy after Safranin O / Fix Green staining (×200). Implementation

[0022] The specific embodiments of the present invention will be further described in detail below with reference to the accompanying drawings and examples. The following examples are only used to more clearly illustrate the technical solutions of the present invention, so that those skilled in the art can better understand and utilize the present invention, and are not intended to limit the scope of protection of the present invention.

[0023] The experimental methods, production processes, instruments, and equipment involved in the embodiments of this invention are all conventional names in the art, and are very clear and distinct in their respective fields of application. Those skilled in the art can understand the conventional process steps and apply the corresponding equipment based on these names, and implement them according to conventional conditions or conditions recommended by the manufacturer.

[0024] The raw materials or reagents used in the embodiments of this invention are not subject to any special restrictions on their source; they are all conventional products that can be purchased commercially. They can also be prepared according to conventional methods well known to those skilled in the art.

[0025] This invention relates to a drug for treating rheumatoid arthritis, the active ingredient of which is compound 2,4',5'-trihydroxy-5,2'-dibromobenzophenone. The drug may also include a common pharmaceutical carrier or excipient used in combination with the active ingredient. The drug can be formulated into common dosage forms such as tablets, injections, pellets, solid dispersions, or capsules, depending on specific requirements. Example

[0026] 1. Laboratory animals

[0027] SD rats, half male and half female, weighing 180 - 220 g. Source of animals: Beijing Vital River Laboratory Animal Technology Co., Ltd., License number: SCXK (Beijing) 2021 - 0006, Certificate number: 110011230107541852.

[0028] All animals used in this experiment and related disposals complied with the requirements of animal welfare. The experiment conducted has passed the ethical review of the Animal Welfare Committee of this institution.

[0029] 2. Source of feed

[0030] Beijing Keao Xieli Feed Co., Ltd.

[0031] 3. Drugs and reagents

[0032] LF1 tablets (Batch number: 20210429), from Shanxi Zhendong Pharmaceutical Co., Ltd); Celecoxib capsules (Batch number: 8142830), from Pfizer Pharmaceuticals Limited.

[0033] Freund’s Adjuvant Complete (Sigma Company, USA); Immunization Grade Bovine Type Ⅱ Collagen, Solution (Chondrex Company, USA).

[0034] Rat MMP - 9 ELISA kit (Wuhan Beinilai Biotechnology Co., Ltd., Batch number: RA20128); Rat IL - 1β ELISA kit (Wuhan Beinilai Biotechnology Co., Ltd., Batch number: RA20020); Rat IL - 17A ELISA kit (Wuhan Beinilai Biotechnology Co., Ltd., Batch number: RA20477); Rat TNF - α ELISA kit (Wuhan Beinilai Biotechnology Co., Ltd., Batch number: RA20035); Rat IgM ELISA kit (Wuhan Beinilai Biotechnology Co., Ltd., Batch number: RA20444); Rat IgG ELISA kit (Wuhan Beinilai Biotechnology Co., Ltd., Batch number: RA20096).

[0035] 4. Preparation of animal model of rheumatoid arthritis

[0036] Healthy adult SD rats (180 - 220 g), half male and half female, were fed with ordinary feed for 1 week for adaptation.

[0037] Primary immunization: Rats were anesthetized with 2.5% tribromoethanol. The anesthetic was administered intraperitoneally at a dose of 5 ml / kg based on the rat's body weight. Once the rats were still, 0.1 ml of the drug was injected subcutaneously at three points on the back, the base of the tail, and the left hind paw, for a total of 0.5 ml of bovine type II collagen emulsion. Booster immunization: One week after the primary immunization, 0.2 ml of bovine type II collagen emulsion was injected intraperitoneally.

[0038] 5. Group and Dosage Design

[0039] Two weeks after the initial immunization, except for the blank group (n=12), the rats that had developed the model were randomly divided into CIA model group, celecoxib positive control group (21.0 mg / kg), LF1 high (45.0 mg / kg), medium (22.5 mg / kg), and low (11.3 mg / kg) dose groups, with 12 rats in each group.

[0040] 6. Administration method

[0041] Drug preparation: LF1 high, medium and low dose groups, celecoxib positive control group, suspended in 0.5% CMC-Na.

[0042] Route of administration: Gavage.

[0043] Dosage volume: The blank group and the model group were given CMC-Na 1.0ml / 100g; the celecoxib group, the high, medium and low dose groups of LF1 were given different concentrations of the drug, and the dosing volume was 1.0ml / 100g.

[0044] Dosage time: Start medication 7 days after booster immunization.

[0045] Dosage frequency: once daily for a total of 6 weeks.

[0046] 7. Experimental Methods

[0047] During the 6-week dosing period following a booster immunization week, rat ankle diameter and body weight were monitored, bone damage was assessed using X-ray, and the rat arthritis index AI was evaluated according to the following criteria.

[0048]

[0049] Six weeks after administration, patients fasted overnight and underwent intraperitoneal anesthesia. Blood was drawn from the abdominal aorta to detect relevant indicators. These indicators included serum levels of MMP-9, IL-17A, IgG, IgM, IL-1β, and TNF-α.

[0050] The spleen was washed with physiological saline, its weight was recorded, and the spleen coefficient was calculated. A portion of the spleen tissue was then immersed in 10% neutral formalin for pathological morphological analysis.

[0051] One ankle joint was taken and fixed by immersing it in 4% paraformaldehyde tissue fixation solution. Safranin O / Fast Green staining was used to detect the degree of cartilage damage and bone erosion in rats.

[0052] 8. Statistical methods

[0053] Data results This indicates that, using GraphPad 9.0 software, one-way ANOVA was used for comparisons among multiple groups, and pairwise comparisons between groups were performed using... t Test, when p A value < 0.05 is considered statistically significant. * , # express p <0.05; ** , ## express p <0.01; *** , ### express p <0.001.

[0054] 9. Results and Analysis

[0055] 1) Effects on rat body weight

[0056]

[0057] Rheumatoid arthritis (RA) can lead to decreased food intake, slow weight gain, or even reduced weight gain in rats. The control group showed a steady increase in weight over time; the CIA model group showed a slight increase in weight over time, but this was significantly lower than the control group; the LF1 and celecoxib groups improved the rats' food intake, resulting in a significant increase in weight compared to the model group; the high-dose LF1 group showed significantly better weight gain after 28 days than the celecoxib group.

[0058] 2) Effect on ankle joint diameter in rats

[0059]

[0060]

[0061] Rheumatoid arthritis primarily affects various joints in rats, causing ankle swelling and a significant increase in ankle joint diameter. In the control group, the ankle joint diameter remained around 7 mm. Compared to the model group, the LF1 administration group and the celecoxib group alleviated the degree of ankle swelling in rats.

[0062] 3) Effect on arthritis index score in rats

[0063]

[0064] The arthritis index, a recognized standard for evaluating the severity of arthritis in animal models of rheumatoid arthritis, gradually increases as the disease progresses. The joints of rats in the control group were normal; compared to the model group, the arthritis index of rats in the LF1 treatment group and the celecoxib group showed a decreasing trend. After 6 weeks of treatment, the high-dose LF1 group showed significantly better results than the celecoxib group.

[0065] 4) Effects on cartilage damage and bone erosion in rats

[0066] Figure 1 X-ray monitoring was used to assess changes in the ankle joints of rats. In the control group, there was no swelling, and the outlines of the ankle and toe joints were clear. In the CIA model group, rats showed significant swelling and deformity, with both the ankle and toe joints exhibiting varying degrees of deformity and covered with erosions. Compared to the model group, the LF1-treated group and the celecoxib group showed significant reductions in joint swelling and ankle / toe joint deformities.

[0067] Figure 2 The pathological comparison of the ankle joint synovium of rats in each group was observed under electron microscopy after Safranin O / Fix Green staining (×200). The cartilage structure in the blank group was clear and intact; the synovial structure of the ankle joint of rats in the CIA model group was destroyed, the cartilage structure was severely damaged, the red part almost disappeared, and the proportion of cartilage area decreased significantly; the degree of cartilage damage in the ankle joint of rats in the LF1 administration group and the celecoxib group was significantly improved.

[0068] 5) Effects on serum levels of MMP-9, IL-17A, IgG, and IgM in rats

[0069]

[0070] MMP-9, IL-17A, IgG, and IgM are diagnostic indicators in the blood of clinical rheumatoid arthritis patients. Compared with the control group, the levels of cytokines MMP-9, IL-17A, IgG, and IgM in the serum of rats in the CIA model group were significantly increased; compared with the CIA model group, the LF1 administration group significantly reduced the levels of these indicators in the serum of rats, and improved the pathological damage to the ankle joint caused by collagen-induced rheumatoid arthritis in rats.

[0071] 6) Effects on the levels of inflammatory factors in rat serum

[0072]

[0073] Rheumatoid arthritis is characterized by synovial inflammation, with a significant increase in serum inflammatory factors. Compared with the control group, the levels of inflammatory factors TNF-α and IL-1β in the serum of rats in the CIA model group were significantly increased. Compared with the model group, the LF1 administration group showed a dose-dependent decrease in the levels of TNF-α and IL-1β in the serum of CIA rats. The high-dose LF1 group was superior to the celecoxib group in reducing the inflammatory factor IL-1β.

[0074] 7) Effect on spleen coefficient in rats

[0075]

[0076] Patients with rheumatoid arthritis often experience splenomegaly due to an overactive autoimmune system. Compared with the control group, the spleen coefficient of rats in the CIA model group was significantly increased; the spleen coefficient of rats in the LF1 administration group was significantly reduced in a dose-dependent manner; the reduction was more pronounced in the high-dose LF1 group than in the celecoxib group.

[0077] 10. Conclusion

[0078] The above results show that 2,4',5'-trihydroxy-5,2'-dibromobenzophenone has a clear anti-rheumatoid arthritis effect in rats, effectively slowing down weight loss, reducing ankle swelling, lowering arthritis scores, alleviating ankle bone erosion in rats, and reducing the levels of MMP-9, IL-17A, IgG, and IgM in rat serum, as well as significantly reducing the levels of IL-1β and TNF-α inflammatory factors in rat serum. The main therapeutic indicators of MMP-9, IL-17A, IL-1β, and IgM are superior to those of celecoxib, a current first-line clinical drug, especially the high-dose group, which shows significant efficacy and demonstrates important application development prospects.

[0079] The above embodiments of the present invention do not describe all details exhaustively, nor do they limit the present invention to the embodiments described above. Various changes, modifications, substitutions, and variations made by those skilled in the art to these embodiments without departing from the principles and spirit of the present invention should be included within the scope of protection of the present invention.

Claims

1. Application of the halogenated phenol compound 2,4',5'-trihydroxy-5,2'-dibromobenzophenone in the preparation of drugs for treating rheumatoid arthritis.

2. An antirheumatoid arthritis drug, wherein the drug contains an effective dose of the compound 2,4',5'-trihydroxy-5,2'-dibromobenzophenone as the active pharmaceutical ingredient.

3. The drug according to claim 2, characterized in that: The drug also includes a common pharmaceutical carrier or excipient used in combination with the active ingredient.

4. The drug according to claim 2, characterized in that... The drug is in the form of tablets, injections, pellets, solid dispersions, or capsules.

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