Application of rusotinib in preparation of medicine for treating osteoarthritis

Abstract

The invention discloses an application of rusotinib in preparation of a medicine for treating osteoarthritis. It is proved for the first time that when rusotinib is injected into an articular cavity or orally taken, the osteoarthritis symptom can be remarkably relieved, meanwhile, metabolism of cartilage cells can be improved, and the progress of arthritis can be slowed down. The rusotinib is used for treating the osteoarthritis, so that the life quality of a patient can be improved, the dependence on analgesics is reduced, the medical cost is reduced, and a new thought is provided for the medicine treatment of the osteoarthritis.

Description

Technical Field

[0001] The present invention relates to the field of biomedical technology, and in particular to the application of ruxolitinib in the preparation of a drug for treating osteoarthritis. Background Art

[0002] Osteoarthritis (OA) is a non-inflammatory degenerative joint disease characterized by joint pain and stiffness, especially after prolonged activity. It is common in people over 50 years old and affects the quality of life of middle-aged and elderly patients to varying degrees.

[0003] Late-stage osteoarthritis often requires surgical treatments such as artificial joint replacement. However, artificial joints have a lifespan problem, and not all joint surgeries can achieve satisfactory results. Therefore, conservative treatment of osteoarthritis to relieve symptoms and prolong the service life of joints is particularly important. Intra-articular drug injection is one of the current treatments for relieving osteoarthritis. Currently commonly used intra-articular drug injections include the use of anesthetic drugs (such as lidocaine), joint lubricating drugs (such as sodium hyaluronate) and glucocorticoids. In addition, platelet-rich plasma (PRP) therapy is also used for intra-articular injection to treat osteoarthritis.

[0004] However, although the combined intra-articular injection of anesthetic drugs (such as lidocaine), joint lubricating drugs (such as sodium hyaluronate) and glucocorticoids can relieve symptoms in the short term, this method cannot delay the progression of the disease. PRP treatment is relatively expensive and requires the extraction of a certain amount of autologous blood, which causes certain damage to the patient. The use of intra-articular injection of drugs has certain limitations, and oral drugs for the treatment of osteoarthritis are more easily accepted by patients.

[0005] Ruxolitinib is a Janus kinase (JAK 1 / JAK2) inhibitor, mainly used to treat adult patients with intermediate-risk or high-risk myelofibrosis (MF), including primary MF, MF after polycythemia vera (PV), and MF after essential thrombocythemia. In addition, ruxolitinib is also used to treat atopic dermatitis, vitiligo and other diseases. In recent years, many studies have also demonstrated its anti-tumor and anti-inflammatory capabilities. However, there are no reports on the use of ruxolitinib in the treatment of osteoarthritis. Summary of the invention

[0006] The purpose of the present invention is to provide a method for preparing a drug for treating osteoarthritis by using ruxolitinib in order to address the deficiencies in the prior art.

[0007] To achieve the above object, the technical solution adopted by the present invention is:

[0008] Provided is the use of ruxolitinib in preparing a drug for treating osteoarthritis.

[0009] Furthermore, the drug for treating osteoarthritis is ruxolitinib injection or ruxolitinib oral agent.

[0010] Furthermore, the ruxolitinib injection comprises ruxolitinib as the only active ingredient, and a solvent.

[0011] More preferably, the solvent is physiological saline.

[0012] Furthermore, the concentration of the ruxolitinib injection is 2.5-3.0 μM.

[0013] The present invention adopts the above technical solution, and has the following technical effects compared with the prior art:

[0014] This invention proves for the first time that intra-articular injection or oral administration of ruxolitinib can significantly relieve the symptoms of osteoarthritis while improving the metabolism of cartilage cells and slowing down the progression of arthritis. Using ruxolitinib to treat osteoarthritis can improve the quality of life of patients, reduce dependence on analgesics, and reduce medical costs, providing a new idea for drug treatment of osteoarthritis. BRIEF DESCRIPTION OF THE DRAWINGS

[0015] Figure 1 Shown are the results of Alcian blue staining of mouse chondrocytes after IL1 and Rux stimulation.

[0016] Figure 2 Shown are the results of histological staining after Rux treatment (injection) in the mouse OA model.

[0017] Figure 3 Shown are the results of gait analysis after Rux treatment (oral) in a mouse OA model.

[0018] Figure 4 Shown are the results of the Von Frey pain sensitivity test in a mouse OA model after Rux treatment (oral administration). DETAILED DESCRIPTION

[0019] The present invention is further described below in conjunction with the accompanying drawings and specific embodiments, but is not intended to be limiting of the present invention. It should be noted that the embodiments and features in the embodiments of the present invention may be combined with each other without conflict.

[0020] Pharmaceuticals and biomaterials:

[0021] Ruxolitinib (Rux), MedChemExpress, Cat:HY-50856;

[0022] Experimental mice: C57BL / 6J, Vitallife;

[0023] Recombinant mouse IL-1β, Suzhou Nearshore Protein Technology Co., Ltd.

[0024] Example 1

[0025] Preparation of 2.5 μM Ruxolitinib Injection:

[0026] Take 1 mg of ruxolitinib powder and dissolve it in 1306 ml of normal saline at room temperature. After the drug is completely dissolved, filter it through a 0.22 μm filter to prepare a sterile 2.5 μM ruxolitinib injection. The injection is stored at -20 degrees to avoid repeated freezing and thawing.

[0027] For oral drug treatment, ruxolitinib was dissolved in DMSO, then ground and mixed with food. In addition to regular food, each mouse was fed a small amount of food containing ruxolitinib (60 mg / kg) or DMSO (0.5 g) every day.

[0028] Example 2

[0029] Cell experiments:

[0030] 1) For newborn mice, cut the knee joint skin after carbon dioxide asphyxiation and directly separate the knee joint cartilage. The separated tissue was washed 5 times with sterile PBS, digested with 0.25% trypsin for 30 minutes, and then replaced with 0.15% type II collagenase for 2 hours. Subsequently, the digestion solution was passed through a 70μm cell sieve, centrifuged and re-selected, and cultured in a DMEM medium containing 10% FBS and 1% double antibody at 37°C and 5% CO2 incubator for routine culture. When the cells are 80-90% attached to the wall, trypsin digestion and subculture are used for subsequent experiments.

[0031] 2) Rux stimulation: 2×10 primary mouse chondrocytes were cultured 4 / well were inoculated in a 12-well plate, and when the cells adhered to the wall and grew to 60-70%, 10 ng / ml IL-1β was added to treat the chondrocytes. The Rux treatment group was treated with 2.5 μM Rux solution at the same time.

[0032] 3) Alcian blue staining: The cell culture plate was fixed with 4% PFA for 15 minutes, then stained with Alcian blue (AB) solution for 15 minutes, washed with distilled water until colorless, and observed under a stereomicroscope. Alcian blue staining showed that the AB staining of chondrocytes after Rux stimulation for 48 hours deepened, indicating that the secretion of extracellular matrix by chondrocytes increased ( Figure 1 ).

[0033] Example 3

[0034] Animal experiment 1:

[0035] The experiment used 10-week-old male wild-type C57 mice. 30 mice were randomly divided into three groups: a sham operation group, a control group and a joint cavity treatment group, with 10 mice in each group. The control group and the joint cavity treatment group were constructed with a mouse OA model by anterior cruciate ligament transection (ACLT) in mice. The sterile 2.5μM ruxolitinib injection configured in Example 1. For the sham operation group and the control group mice, the knee joint injection point was locally disinfected and the needle was inserted into the joint cavity, and 5μL of normal saline was injected into the joint cavity every other day; for the joint cavity treatment group mice, the knee joint injection point was locally disinfected and the needle was inserted into the joint cavity, and 5μL of 2.5μM ruxolitinib injection was injected into the joint cavity every other day, and the local disinfection was performed after the needle was withdrawn. After 4 weeks, the mice were killed by carbon dioxide asphyxiation, and their knee joint tissues were isolated and fixed in 4% PFA solution for 24 hours, decalcified in 10% ethylenediaminetetraacetic acid for 1 week, and then dehydrated in graded ethanol and embedded in paraffin. The sections were cut into 5 μm thick sections, and the degeneration of the mouse knee cartilage was evaluated by safranin O-fast green staining (SO-FS). By SO-FS staining, it was observed that the severity of OA in Rux-treated mice was significantly milder than that in the control group ( Figure 2 ), suggesting its role in treating OA.

[0036] Animal experiment 2:

[0037] The experiment used 10-week-old male wild-type C57 mice. 30 mice were randomly divided into three groups: a sham operation group, a control group and an oral treatment group, with 10 mice in each group. The control group and the treatment group established a mouse OA model by anterior cruciate ligament transection (ACLT) in mice. The mixture of ruxolitinib and food configured in Example 1. For the sham operation group and the control group mice, they were fed a small amount of food containing DMSO (0.5g) every day; for the oral treatment group mice, each mouse was also fed a small amount of food containing ruxolitinib (60mg / kg body weight) (0.5g) every day. After 4 weeks, the mouse behavioral analysis was performed, and the mouse gait analysis ( Figure 3 ) and Von Frey pain sensitivity experiment ( Figure 4 ) found that mice in the oral drug group showed behavioral improvements in osteoarthritis, proving that the pain symptoms caused by osteoarthritis were alleviated.

[0038] The above description is only a preferred embodiment of the present invention, and does not limit the implementation mode and protection scope of the present invention. For those skilled in the art, it should be aware that all solutions obtained by equivalent substitutions and obvious changes made using the contents of the specification and illustrations of the present invention should be included in the protection scope of the present invention.

Claims

1. Application of ruxolitinib in the preparation of drugs for the treatment of osteoarthritis.

2. The use according to claim 1, characterized in that: The drug for treating osteoarthritis is ruxolitinib injection or ruxolitinib oral preparation.

3. The use according to claim 2, characterized in that: The ruxolitinib injection comprises ruxolitinib, the only active ingredient, and a solvent.

4. The use according to claim 3, characterized in that: The solvent is physiological saline.

5. The use according to claim 2, characterized in that: The concentration of the ruxolitinib injection is 2.5-3.0 μM.

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