Use of soluble lanthanum in ameliorating arthritis

Abstract

The present invention relates to the use of soluble lanthanum in ameliorating arthritis, belongs to the technical field of rheumatoid arthritis treatment, and provides a drug or preparation containing soluble lanthanum, wherein the usage amount of the soluble lanthanum in the drug or preparation is 0.01-10 ng / kg. The soluble lanthanum at the dose provided can make the inflammatory factor level of rats suffering from rheumatoid arthritis close to that of rats not suffering from rheumatoid arthritis, significantly alleviating the synovial membrane inflammation of rheumatoid arthritis model rats, with a wide dosage range. HE-stained sections of the ankle joint show that injected doses of the soluble lanthanum can significantly alleviate the bone injury of the rheumatoid arthritis model rats. No toxic and side effects are observed, indicating that the soluble lanthanum can effectively ameliorate the rheumatoid arthritis pathology.

Description

Application of soluble lanthanum in improving arthritis Technical Field

[0001] The present invention relates to the technical field of rheumatoid arthritis treatment, and in particular to an application of soluble lanthanum in improving arthritis. Background Art

[0002] Rheumatoid arthritis (RA), commonly known as the "immortal cancer," is considered the world's leading cause of disability. It is a chronic, age-related, systemic autoimmune disease of unknown etiology, primarily characterized by inflammatory synovitis. The underlying pathology is synovial cell proliferation, thickening of the lining, infiltration of multiple inflammatory cells, pannus formation, and destruction of cartilage and bone tissue. The main clinical manifestations are recurrent, erosive, symmetrical polyarthritis and swelling of the small joints of the hands and feet, ultimately leading to joint deformity and loss of function, severely impacting patients' quality of life. Specific clinical manifestations include:

[0003] (1) Multiple joints are affected. The affected joints include the hands, feet, wrists, ankles, and temporomandibular joints. Other joints may include the elbows, shoulders, cervical spine, hips, and knees. The joints are symmetrical and often involve ≥5 joints. (2) Joint deformities. Hand deformities include spindle-shaped swelling, ulnar deviation, swan neck deformity, and buttonhole deformity. Foot deformities include calcaneal deformity caused by inferior subluxation of the metatarsal head, valgus deformity, metatarsophalangeal joint subluxation, hammer toes, and valgus deformity.

[0004] (3) Morning stiffness, inflexible joint movement when getting up in the morning, the duration of which is proportional to the severity of the inflammation. (4) Fever, rheumatoid nodules, rheumatoid vasculitis, lymphadenopathy, neck pain, neck weakness and difficulty maintaining its normal position, atlantoaxial subluxation.

[0005] (5) If not treated promptly and effectively, RA may lead to systemic inflammation, affecting multiple organs / systems, leading to abnormalities in heart, liver, intestinal and muscle function, and in some cases, cognitive decline. Heart involvement may include pericarditis, pericardial effusion, nodules of the epicardium, myocardium and valves, myocarditis, coronary artery inflammation, aortitis, conduction disorders, chronic endocarditis and valvular fibrosis; respiratory system involvement may include pleurisy, pleural effusion, pulmonary arteritis, interstitial lung disease, sarcoidosis, etc.; nervous system, in addition to symptoms of peripheral nerve compression, may also induce neurological diseases, myelopathy, peripheral neuropathy, ischemic neuropathy secondary to vasculitis, muscle hypertrophy and drug-induced nervous system lesions; eyes, young patients may have uveitis, adults may have scleritis, and may also have dry conjunctivitis, scleromalacia, scleromalacia perforation, corneal dissolution; kidney damage, anemia, etc.

[0006] Although the pathogenesis of RA is still unclear, abnormal cellular immune function and macrophage hyperresponsiveness are considered to be key factors in the development of the disease. Currently, the commonly used drugs for the treatment of RA include NSAIDs, GCs, DMARDs, biological products and Chinese herbal medicines. The mechanism of action of chemical drugs and biological products for the treatment of RA is mainly related to the inhibition of target enzymes, inflammatory factors and the activity of inflammatory signaling pathways. However, these drugs have certain toxic side effects and limitations, which to a certain extent limit their clinical use.

[0007] Lanthanum is a metallic rare earth element with the chemical symbol La. It has two natural isotopes: lanthanum-139 and radioactive lanthanum-138. The half-life of radioactive lanthanum-138 is 1.1×10 11 Lanthanum's content in the Earth's crust was 0.00183%, ranking 28th among the elements in the Earth's crust. It is one of the more abundant rare earth elements, second only to cerium. Lanthanum is chemically active, readily oxidizing in air and combusting upon heating to form oxides and nitrides. It reacts directly with carbon, nitrogen, boron, selenium, silicon, phosphorus, sulfur, and halogen lamps. Lanthanum itself is not particularly toxic to humans, but it exhibits some antimicrobial activity. Summary of the Invention

[0008] The purpose of the present invention is to provide an application of soluble lanthanum in improving arthritis, so as to contribute to solving the problem that current therapeutic drugs for rheumatoid arthritis generally have toxic side effects and limitations.

[0009] To achieve the above objectives, the present invention provides a drug or preparation containing soluble lanthanum, wherein the amount of soluble lanthanum used in the drug or preparation is 0.01-10 ng / kg.

[0010] Preferably, the soluble lanthanum is lanthanum chloride, lanthanum sulfate, or a combination of both.

[0011] A use of the above-mentioned medicine or preparation containing soluble lanthanum in improving arthritis, wherein the use is in improving synovitis and bone damage caused by rheumatoid arthritis.

[0012] Preferably, the improvement of synovitis caused by rheumatoid arthritis is the improvement of rheumatoid arthritis synovial lesions.

[0013] Preferably, the improvement of bone damage is the improvement of cartilage damage induced by rheumatoid arthritis.

[0014] Therefore, the application of soluble lanthanum in improving arthritis provided by the present invention has the following specific technical effects:

[0015] (1) Through the administration experiment on the established CIA rheumatoid arthritis rat model, it was shown that the injection of the soluble lanthanum provided by the present invention can make the inflammatory factor level of rats suffering from rheumatoid arthritis close to the level of rats without rheumatoid arthritis, indicating that the injection of the soluble lanthanum at the dose of the present invention can significantly alleviate the inflammatory response of rats with rheumatoid arthritis model;

[0016] (2) Through drug administration experiments on the established CIA rheumatoid arthritis rat model, there was no significant difference in the levels of inflammatory factors in the model rats injected with 0.1ng / kg, 0.3ng / kg and 1ng / kg doses, indicating that low concentrations of soluble lanthanum can play a role and rheumatoid arthritis rats have a relatively wide tolerance range for soluble lanthanum;

[0017] (3) Through drug administration experiments on the established CIA rheumatoid arthritis rat model, HE-stained sections of the ankle joints showed that the injected dose of soluble lanthanum can significantly alleviate bone damage in rheumatoid arthritis model rats;

[0018] (4) Through the drug administration experiment on the established CIA rheumatoid arthritis rat model, the model rats after tail vein injection appeared quieter, had a stronger appetite, and had significantly reduced surface pain compared with the rats in the blank model group. Compared with normal rats, no abnormal state was observed / detected in the model rats after 6 weeks of drug administration, indicating that the experimental dose of soluble lanthanum of the present invention is safe, indicating that soluble lanthanum can effectively improve rheumatoid arthritis lesions and can be used as a new potential drug for improving / treating rheumatoid arthritis lesions.

[0019] The technical solution of the present invention is further described in detail below through the accompanying drawings and embodiments. BRIEF DESCRIPTION OF THE DRAWINGS

[0020] In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the following briefly introduces the drawings required for use in the description of the embodiments of the present invention. Obviously, the drawings described below are only some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying any creative labor.

[0021] FIG1 is a photograph of the left foot of a CIA rheumatoid arthritis rat model and a normal rat in Example 1 of the present invention, wherein the Control is a photograph of the left foot of a normal rat, and the Model is a photograph of the left foot of a model rat;

[0022] Figure 2 is a graph of the serum inflammatory cytokine levels of rats in each group 6 weeks after the tail vein injection of lanthanum chloride in Example 1 of the present invention, wherein Control is a blank control rat injected with normal saline at a setting of 1 mL / kg / 3d, and Model is a model rat injected with normal saline at a setting of 1 mL / kg / 3d; LaCl3 (0.1 ng / kg) is a model rat injected with the lanthanum chloride preparation prepared in Example 2 at a setting of 1 mL / kg / 3d (the amount of lanthanum chloride is 0.1 ng / kg); LaCl3 (0.3 ng / kg) is a model rat injected with the lanthanum chloride preparation prepared in Example 2 at a setting of 1 mL / kg / 3d (the amount of lanthanum chloride is 0.3 ng / kg); LaCl3 (1 ng / kg) is a model rat injected with the lanthanum chloride preparation prepared in Example 2 at a setting of 1 mL / kg / 3d (the amount of lanthanum chloride is 1 ng / kg);

[0023] Figure 3 is a HE staining of the ankle joints of rats in each group after 6 weeks of tail vein injection of lanthanum chloride in Example 2 of the present invention, wherein Control is a blank control rat injected with normal saline at a setting of 1 mL / kg / 3d, and Model is a model rat injected with normal saline at a setting of 1 mL / kg / 3d; LaCl3 (1 ng / kg) is a model rat injected with the lanthanum chloride preparation prepared in Example 2 (the amount of lanthanum chloride used is 1 ng / kg) at a setting of 1 mL / kg / 3d; 4× is an image captured under a 4x objective lens, with a scale of 500 μm, 10× is an image captured under a 10x objective lens, with a scale of 200 μm, and 40× is an image captured under a 40x objective lens, with a scale of 50 μm;

[0024] FIG4 is a diagram showing the changes in body weight of rats at different dosages in Example 3 of the present invention (Part A) and the results of ALT, AST, Scr, and BUN level tests (Part B);

[0025] FIG5 is a photograph of kidney tissue sections (Part A) and liver tissue sections (Part B) of rats administered with different dosages in Example 3 of the present invention;

[0026] FIG6 is a blood routine test result of rats treated with different dosages in Example 3 of the present invention. DETAILED DESCRIPTION

[0027] The technical solution of the present invention is further described below with reference to the accompanying drawings and embodiments.

[0028] In order to make the purpose, technical solutions and advantages of the present application clearer, more thorough and more complete, the technical solutions of the present invention are clearly and completely described below through the accompanying drawings and Examples. The following detailed description is an explanation of the embodiments and is intended to provide further details of the present invention. Unless otherwise specified, all technical terms used in the present invention have the same meaning as those generally understood by those skilled in the art to which the application belongs.

[0029] The instruments, equipment, reagents and materials used in the examples were obtained from commercial sources.

[0030] Example 1

[0031] The CIA rheumatoid arthritis rat model was established as follows:

[0032] Wistar rats (clean grade, male, weighing 200 g ± 20 g) were purchased from Beijing Weitonglihua Experimental Animal Technology Co., Ltd. They were housed in the rat breeding room of the SPF barrier area of ​​the Animal Experimental Center of Inner Mongolia Medical University at a temperature of 18-22°C and a humidity of 45-60%. The animals received food and water on their own initiative.

[0033] After one week of acclimatization, all rats except the control rats were injected with 0.1 mL of a suspension of bovine type II collagen (Chondrex, 20022, USA) and complete Freund's adjuvant (F5881-10 mL, Sigma-Aldrich) at the base of the tail and the toes. Seven days later, a booster injection of 0.1 mL of the same suspension was administered to the toes, thereby inducing a rat model of rheumatoid arthritis (CIA). The animals were allowed to consume food and water ad libitum. Figure 1 shows photographs of the left feet of rats with and without CIA. The control rats show photographs of the left feet of normal rats, while the model rats show photographs of the left feet of model rats. As shown in Figure 1, the entire foot (including the ankle, foot, and toes) of the rats with CIA showed significant erythema and severe swelling compared to the normal rats, indicating that the CIA rat model was successfully established.

[0034] Example 2

[0035] A preparation comprising lanthanum chloride is prepared as follows:

[0036] Use an analytical balance to accurately weigh 1 mg of LaCl3 standard (LaCl3, produced by Sigma, Germany, batch number 298182, chromatographic purity 99.9%) in a dry environment at room temperature, add it to 50 mL sterile centrifuge tube No. ①, then add 50 mL of normal saline to fully dissolve the standard. At this time, the LaCl3 concentration is 20 μg / mL; draw 1 mL of the solution from centrifuge tube No. ① into 50 mL sterile centrifuge tube No. ②, add normal saline to dilute to 50 mL, at this time, the LaCl3 concentration is 40 ng / mL; draw the solution from centrifuge tube No. ② Transfer 1 mL of the solution from tube 3 to a 50 mL sterile centrifuge tube and add saline to 40 mL to create a 1 ng / mL LaCl₃ solution. Pipette 3 mL of the solution from tube 3 to a 50 mL sterile centrifuge tube 4 and add saline to 10 mL to create a 0.3 ng / mL LaCl₃ solution. Pipette 1 mL of the solution from tube 3 to a 50 mL sterile centrifuge tube 5 and add saline to 10 mL to create a 0.1 ng / mL LaCl₃ solution. Pass the prepared LaCl₃ solution through a 0.45 μm filter using a 5 mL syringe and inject into the rat's tail vein.

[0037] Example 3

[0038] The CIA rheumatoid arthritis rat model constructed in Example 1 was used for drug administration experiments, and the method was as follows:

[0039] The rats that successfully established the model in Example 1 were randomly divided into four groups. One group was randomly selected and injected with the lanthanum chloride preparation prepared in Example 2 at a dose of 1 ng / kg / 3 d through the tail vein, designated as the LaCl3 (1 ng / kg) group; one group was randomly selected and injected with the lanthanum chloride preparation prepared in Example 2 at a dose of 0.3 ng / kg / 3 d, designated as the LaCl3 (0.3 ng / kg) group; one group was randomly selected and injected with the lanthanum chloride preparation prepared in Example 2 at a dose of 0.1 ng / kg / 3 d, designated as the LaCl3 (0.1 ng / kg) group; the remaining group was injected with normal saline at a dose of 1 mL / kg, designated as the blank model group (Model group), with eight rats in each group successfully establishing the model. Eight normal / healthy rats were randomly selected and injected with normal saline at a dose of 1 mL / kg, designated as the control group (Control group).

[0040] Effect Example 1

[0041] Investigate the levels of inflammatory factors in rats

[0042] Six weeks after administration, the rats were sacrificed and blood was collected from the abdominal aorta. Serum samples were obtained using a high-speed refrigerated centrifuge (Sigma, Germany). Serum levels of IL-10, IL-13, IL-6, and TNF-α were measured using an Infinite M200pro microplate reader (Tecan, Switzerland). The results are shown in Table 1. Data were processed using the statistical software package SPSSI 3.0, and one-way ANOVA was used for analysis of variance. When variances were homogeneous, the LSD test was used for multiple comparisons between groups; when variances were unequal, the Welch test with approximate ANOVA was used. Data were plotted using Sigma Plot 12.0 software (Figure 2). Statistical significance was considered when P < 0.05.

[0043] Table 1

[0044] ;

[0045] In the table, # indicates P < 0.05, ## indicates P < 0.01, and ### indicates P < 0.001 compared with the control group; * indicates P < 0.05, ** indicates P < 0.01, and *** indicates P < 0.001 compared with the blank model group

[0046] As shown in Table 1 and Figure 2, compared with the control group, the serum levels of the proinflammatory cytokines TNF-α and IL-6 increased in the blank model group (Model group). However, the TNF-α and IL-6 levels decreased in the LaCl3 (0.1 ng / kg), LaCl3 (0.3 ng / kg), and LaCl3 (1 ng / kg) treatment groups, approaching those in the control group. The serum levels of the proinflammatory cytokines IL-10 and IL-13 decreased in the control group (Control group) compared with the blank model group. However, the IL-10 and IL-13 levels increased in the LaCl3 (0.1 ng / kg), LaCl3 (0.3 ng / kg), and LaCl3 (1 ng / kg) treatment groups, approaching those in the control group. These results indicate that LaCl3 significantly alleviates the inflammatory response in rats with rheumatoid arthritis.

[0047] Effect Example 2

[0048] Observation of rat joint tissue pathological sections

[0049] The ankle joints of rats were fixed with 4% paraformaldehyde for 48 hours, and then pathological sections of the joint tissues were made. The sections were then stained with hematoxylin-eosin (HE) and observed under a microscope. The results are shown in Figure 3.

[0050] As can be seen from Figure 3, the synovial membrane of the ankle joints of rats in the blank model group (Model group) showed obvious inflammatory cell infiltration, visible pannus structure, and obvious damage to the cartilage tissue. After treatment with LaCl3 (1 ng / kg), the inflammatory cell infiltration of the synovial membrane was alleviated and the cartilage surface was smooth, indicating that LaCl3 can significantly reduce synovial hyperplasia and cartilage erosion in rats with rheumatoid arthritis.

[0051] Effect Example 3

[0052] Conduct acute toxicity testing

[0053] The model rats after tail vein injection appeared calmer and had a stronger appetite than the rats in the blank model group, indicating that pain was significantly alleviated. Compared with normal rats, no abnormal conditions were observed / detected in the model rats after 6 weeks of administration, indicating that the experimental dose of soluble lanthanum of the present invention is safe.

[0054] In addition, 50 Wister rats, half male and half female, were selected and continuously administered 0.1 ng / kg, 0.3 ng / kg, 1 ng / kg, 3 ng / kg, and 9 ng / kg LaCl3 into the tail vein for 15 days, once every three days.

[0055] (1) The changes in body weight of rats in each LaCl3-administered group were examined. The results are shown in Part A of Figure 4. As can be seen from Part A of Figure 4, the body weight of rats in the groups administered with 0.1 ng / kg, 0.3 ng / kg, and 1 ng / kg LaCl3 per day increased with the increase in the number of experimental days, while the body weight of rats in the groups administered with 3 ng / kg and 9 ng / kg LaCl3 per day began to decrease significantly on the 6th day of administration (P < 0.01).

[0056] (2) After 15 days of administration, serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) levels in each group of rats were measured to evaluate the effect of lanthanum chloride on liver function, and creatinine (Scr) and blood urea nitrogen (BUN) levels were measured to evaluate the effect of lanthanum chloride on renal function. The results are shown in Figure 4, Part B. Lanthanum chloride caused an increase in ALT and AST levels in the rat serum, but the increase was not significant. There was no significant effect on the kidneys either.

[0057] (3) To further evaluate whether trace lanthanum chloride produced toxic effects on the kidneys and livers of rats in the short term and to preliminarily confirm the safe dose range, the present invention used HE staining to perform histopathological analysis on the livers and kidneys of rats after 15 days of administration. The results are shown in FIG5 , where Part A is a section of rat kidney tissue and Part B is a section of rat liver tissue. As can be seen from Part A of FIG5 , short-term administration of trace lanthanum chloride did not cause obvious histopathological changes in the kidney tissue of rats. As can be seen from Part B of FIG5 , at a dose of 9 ng / kg, the liver tissue of rats showed inconsistent size, disordered arrangement, and inflammatory cell infiltration. Combined with the results of Part B of FIG4 , it can be seen that a dose of 9 ng / kg would cause a certain degree of damage to the liver of rats.

[0058] (4) To further determine the short-term toxicity of intravenous trace lanthanum chloride, the present invention collected venous blood from the retinal venous plexus of rats after completing a 15-day dosing cycle and performed a routine blood test. The test results are shown in FIG6 . As can be seen from the figure, compared with the other groups, the rats in the 9 ng / kg group had a decrease in red blood cell count, hemoglobin level, and mean hemoglobin level. This indicates that the rats in the 9 ng / kg group may have mild anemia.

[0059] The results show that the soluble lanthanum in the dosage provided by the present invention has no obvious toxicity to experimental rats.

[0060] Therefore, the soluble lanthanum at the dose provided by the present invention can significantly alleviate the inflammatory response of rats with rheumatoid arthritis model. The soluble lanthanum at doses of 0.1 ng / kg, 0.3 ng / kg, and 1 ng / kg has no obvious toxicity and a wide dosage range of application. HE-stained sections of ankle joints show that the injected dose of soluble lanthanum can significantly alleviate bone damage in rats with rheumatoid arthritis model. No toxic side effects were observed, indicating that soluble lanthanum can effectively improve rheumatoid arthritis lesions and can be used as a new potential drug for improving / treating rheumatoid arthritis lesions.

[0061] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention rather than to limit the same. Although the present invention has been described in detail with reference to the preferred embodiments, those skilled in the art should understand that they can still modify or replace the technical solutions of the present invention with equivalents, and these modifications or equivalent replacements cannot cause the modified technical solutions to deviate from the spirit and scope of the technical solutions of the present invention.

Claims

1. A drug or preparation containing soluble lanthanum, characterized in that: The dosage of soluble lanthanum in the drug or preparation is 0.01 - 10 ng / kg.

2. A drug or preparation containing soluble lanthanum according to claim 1, characterized in that: The soluble lanthanum is one or a combination of two of lanthanum chloride and lanthanum sulfate.

3. Use of a drug or preparation containing soluble lanthanum as described in claim 1 or 2 in improving arthritis, characterized in that: The application is for improving synovitis and bone injury caused by rheumatoid arthritis.

4. Use of a drug or preparation containing soluble lanthanum according to claim 3 in improving arthritis, characterized in that: The improvement of synovitis caused by rheumatoid arthritis is to improve the synovial lesions of rheumatoid arthritis.

5. Use of a drug or preparation containing soluble lanthanum according to claim 3 in improving arthritis, characterized in that: The improvement of bone injury is to improve the cartilage injury induced by rheumatoid arthritis.

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