Application of YE6144 in the preparation of drugs for treating skeletal muscle atrophy caused by breast cancer

Abstract

The present invention relates to the application of YE6144 in the preparation of a drug for treating skeletal muscle atrophy caused by breast cancer, and belongs to the field of biomedicine technology. 21 H 27 ClFN7O, molecular weight is 447.94. The present invention first discovered that the small molecule inhibitor YE6144 can improve skeletal muscle atrophy caused by breast cancer. The experimental results show that YE6144 can significantly improve the symptoms of skeletal muscle atrophy in breast cancer mouse models and improve the skeletal muscle mass of mice. This discovery provides a new treatment strategy for breast cancer patients and is expected to improve the muscle atrophy of patients.

Description

Technical Field

[0001] The invention belongs to the technical field of biomedicine, and specifically relates to the application of YE6144 in preparing a drug for treating skeletal muscle atrophy caused by breast cancer. Background Art

[0002] Skeletal muscles account for approximately half of the human body mass and play a key role in maintaining posture and movement. Not only are they responsible for movement, but they also participate in autocrine, paracrine, and endocrine functions by secreting various signaling molecules, and are essential for maintaining metabolic balance throughout the body. Therefore, maintaining the health of skeletal muscles is very important for a healthy lifestyle. Unfortunately, most patients with advanced cancer may develop cachexia, which is characterized by a continuous loss of skeletal muscle mass, which further leads to muscle weakness and systemic metabolic problems, thereby worsening the prognosis.

[0003] Breast cancer patients often suffer from muscle weakness, which not only increases the risk of death but also reduces the quality of life. Even in the absence of obvious muscle mass loss, breast cancer patients may experience muscle dysfunction. Therefore, it is very urgent to develop some more effective drugs to improve the skeletal muscle mass of patients and improve their quality of life.

[0004] YE6144 is a typical interferon regulatory factor 5 (IRF5) inhibitor that selectively inhibits IRF5 activity by inhibiting IRF5 phosphorylation. Studies have shown that YE6144 has low toxicity. In the autoimmune disease systemic lupus erythematosus (SLE), the small molecule inhibitor YE6144 can selectively inhibit the activity of IRF5, reduce the production of type I interferon, and thus inhibit the progression of SLE in mouse models. So far, there have been no reports on the use of the small molecule inhibitor YE6144 in the treatment of skeletal muscle atrophy caused by breast cancer. Summary of the invention

[0005] The purpose of the present invention is to solve the deficiencies of the prior art and provide the use of YE6144 in the preparation of a drug for treating skeletal muscle atrophy caused by breast cancer.

[0006] To achieve the above purpose, the technical solution adopted by the present invention is as follows:

[0007] Application of YE6144 in the preparation of drugs for treating skeletal muscle atrophy caused by breast cancer.

[0008] Preferably, the drug further comprises a pharmaceutically acceptable excipient.

[0009] Preferably, the medicine is in the form of tablets, capsules, granules, powders, pills or injections.

[0010] A drug for treating skeletal muscle atrophy caused by breast cancer, comprising an active ingredient YE6144.

[0011] Small molecule inhibitor YE6144, molecular formula C 21 H 27 ClFN7O, molecular weight 447.94Da.

[0012] The present invention has found that by injecting the YE6144 small molecule inhibitor in vitro, recording the body weight and average food intake of each group of mice, testing the mouse grip strength, and calculating the hematoxylin-eosin staining and cross-sectional area (CSA) of the mouse gastrocnemius muscle, it was found that the small molecule inhibitor YE6144 can alleviate skeletal muscle atrophy caused by breast cancer.

[0013] Compared with the prior art, the present invention has the following beneficial effects:

[0014] The present invention first discovered that the small molecule inhibitor YE6144 can improve skeletal muscle atrophy caused by breast cancer. Experimental results show that YE6144 can significantly improve the symptoms of skeletal muscle atrophy in breast cancer mouse models, improve the skeletal muscle mass of mice, and enhance exercise capacity. The present invention provides YE6144 as an inhibitor for improving skeletal muscle atrophy caused by breast cancer. BRIEF DESCRIPTION OF THE DRAWINGS

[0015] Figure 1 Flow chart of group experiment processing for the study of skeletal muscle atrophy in mice;

[0016] Figure 2 The figure is the result of monitoring the weight of mice; a is the percentage of the weight of mice in each group to the initial weight; b is the cumulative food intake of mice in each group;

[0017] Figure 3 This is the result of the mouse grasping force test;

[0018] Figure 4 Figure 1 is the appearance and weight of the gastrocnemius muscle of mice; a is a representative image of the gastrocnemius muscle of mice in each group; b is the weight of the gastrocnemius muscle of mice in each group;

[0019] Figure 5 The results of hematoxylin-eosin staining of gastrocnemius muscle; a is a representative picture of hematoxylin-eosin staining of gastrocnemius muscle of each group of mice; b is a statistical diagram of the cross-sectional area of ​​gastrocnemius muscle fibers; the scale bar is 20 μm, **P<0.01, ***P<0.001, ****P<0.0001. DETAILED DESCRIPTION

[0020] The present invention is further described in detail below in conjunction with embodiments.

[0021] Those skilled in the art will appreciate that the following examples are only used to illustrate the present invention and should not be considered to limit the scope of the present invention. If no specific techniques or conditions are specified in the examples, the techniques or conditions described in the literature in the art or the product specifications are used. If the manufacturer of the materials or equipment used is not specified, they are all conventional products that can be purchased.

[0022] Example 1 Study on skeletal muscle atrophy in mice treated with YE6144 after 4T1 tumor treatment

[0023] (A) Hematoxylin-eosin staining was used to detect the level of skeletal muscle atrophy in mice.

[0024] 1. Selection of experimental animals and feeding conditions:

[0025] SPF grade 8-week-old BALB / c mice, weighing 16g-18g, were purchased from Jiangsu Jicui Yaokang Biological Co., Ltd. and were raised in an SPF environment with 12 hours of light and 12 hours of darkness, temperature controlled at 18-23°C, and humidity maintained at 40%-60%, and were fed normal mouse chow.

[0026] 2. Experimental methods:

[0027] 2.1 Method for obtaining 4T1 tumor cell suspension

[0028] 4T1 cells were obtained from the Shanghai Cell Bank of the Chinese Academy of Sciences. 4T1 cells were cultured in a DMEM high-glucose medium containing 10% FBS by volume at 37°C in an incubator with 5% CO2 by volume. When the cell density reached 70%-80%, the medium was replaced with serum-free DMEM medium. After 24 hours of culture, the cell culture medium was carefully transferred from the culture dish to a 15 mL centrifuge tube using a pipette. The 15 mL centrifuge tube was centrifuged at 1000 rpm and room temperature for 10 minutes to remove cell debris and obtain a 4T1 tumor cell suspension.

[0029] Among them, the manufacturer and model of FBS is Viva Cell 2910-0500; the manufacturer and model of DMEM high-glucose culture medium is Viva Cell C3113-0500.

[0030] 2.2 Model construction

[0031] After one week of adaptive feeding, the mice were randomly divided into blank control group, 4T1 group, and 4T1+YE6144 group, with 6 mice in each group.

[0032] The 4T1 tumor model was constructed as follows: 8-week-old BALB / c mice were intraperitoneally injected with 4T1 tumor cell suspension one week in advance, with an injection volume of 1×10 6 mL -1 / 0.1mL / mouse. Then the group test treatment was carried out;

[0033] 4T1 group: One week after the injection of 4T1 tumor cell suspension, 100 μL of PBS was injected per animal every four days; (PBS injection was used as a control for YE6144)

[0034] 4T1+YE6144 group: One week after the injection of 4T1 tumor cell suspension, YE6144 was injected at a dose of 10 mg / kg / 100 μL per mouse, once every four days;

[0035] In addition, 8-week-old BALB / c mice that did not receive any experimental treatment were selected as the blank control group, which served as a benchmark for evaluating the effects of the experimental intervention;

[0036] The experimental process is as follows Figure 1 shown.

[0037] (2) Blood samples and skeletal muscle samples were collected from mice on day 28. A portion of the skeletal muscle tissue was embedded in an embedding agent, cut into 7 μM thick slices, and stained with hematoxylin-eosin to detect skeletal muscle atrophy in mice. The remaining tissue was quickly frozen in liquid nitrogen and stored at -80°C for subsequent experiments.

[0038] Among them, the hematoxylin-eosin staining method is as follows:

[0039] In the first step, take the slices out of the -80°C refrigerator, place them at room temperature for 30 minutes, and preheat the hematoxylin in a 60°C water bath.

[0040] In the second step, 4% paraformaldehyde was added dropwise onto the slice surface and fixed for 10 min.

[0041] In the third step, add the preheated hematoxylin in the first step to the surface of the slice and incubate for 10 minutes. Rinse with running water for 5 seconds, and add 1% hydrochloric acid ethanol to differentiate for 30 seconds. Rinse with running water for 5 seconds to terminate the differentiation, and place the slice in 50℃ warm water for 30 seconds to turn blue. Wipe the surface of the slice dry, add 200μL eosin, and incubate for 2 minutes. Rinse with running water for 5 seconds, and dehydrate in 80% ethanol for 1 minute. Use 95% ethanol and anhydrous ethanol for 2 minutes and 4 minutes respectively. Place the slice in xylene for 5 minutes to make it transparent.

[0042] The fourth step was to seal the slides with neutral gum and then observe and photograph them under an upright optical microscope. Image J software was used to statistically process the images and calculate the cross-sectional area of ​​the gastrocnemius muscle fibers.

[0043] 2.2 Grip test

[0044] The gripping force test of mice was carried out using a gripping force instrument purchased from Nanjing Calvin Biological Company. Five control mice, five 4T1 mice, and six 4T1+YE6144 mice were subjected to gripping force adaptation for five days. After the 26th day of the experiment, the three groups of mice that had adapted to gripping force were subjected to gripping force tests. During the test, the mice were made to grasp the iron bar with their four limbs, and the tail of the mouse was grasped by hand and pulled back. The peak gripping force was recorded and repeated five times. The maximum and minimum values ​​were removed and the average was calculated.

[0045] 3. Experimental results:

[0046] (1) Mouse weight monitoring

[0047] The experimental results are as follows Figure 2 As shown. In terms of physiological indicators, compared with the blank control group mice, the weight of the 4T1 group mice was significantly reduced, and the average food intake decreased, indicating that the 4T1 tumor affects the mice's feeding and thus their weight. After the simultaneous injection of YE6144, the 4T1+YE6144 group can significantly improve the weight loss caused by the tumor. This shows that the injection of YE6144 can improve the mice's food intake and increase their weight.

[0048] (2) Mouse grip strength test

[0049] The experimental results are as follows Figure 3 As shown. In terms of motor ability, the gripping force of mice in the 4T1 group was significantly reduced compared with the blank control group, indicating that 4T1 tumors can affect the gripping force of mice. After the simultaneous injection of YE6144, the 4T1+YE6144 group can significantly improve the gripping force of mice. This shows that the injection of YE6144 can improve the motor ability after the injection of 4T1 tumor cell suspension.

[0050] (3) Hematoxylin-eosin staining results

[0051] The experimental results are as follows Figure 4 and Figure 5 shown.

[0052] The results of hematoxylin-eosin staining showed that the skeletal muscle morphology of mice in the blank control group was normal, without obvious damage and the appearance of central nuclei, while the skeletal muscle of mice in the 4T1 group was disordered, with obvious damage and the appearance of central nuclei. After the injection of YE6144, the 4T1+YE6144 group could improve the skeletal muscle disorder caused by 4T1 tumor cell suspension.

[0053] The cross-sectional area of ​​muscle fibers was calculated and it was found that the average cross-sectional area of ​​muscle fibers in the 4T1 group of mice decreased significantly, and the cross-sectional area was between 500 and 2000 μm. 2 Increased small muscle fibers in the range of 3000-4500 μm 2In the 4T1+YE6144 group, it was found that after injection of YE6144, the size of the larger muscle fibers increased by 3000-4500 μm. 2 Larger muscle fibers within the range, improving skeletal muscle atrophy.

[0054] The above experimental results show that YE6144 can significantly improve 4T1 tumor-induced skeletal muscle atrophy in mice.

[0055] Based on this, the present invention provides the use of YE6144 in preparing a drug for treating skeletal muscle atrophy caused by breast cancer.

[0056] The medicine also includes pharmaceutically acceptable excipients.

[0057] The medicine is in the form of tablets, capsules, granules, powders, pills or injections.

[0058] Also provided is a drug for treating skeletal muscle atrophy caused by breast cancer, wherein the active ingredient includes YE6144.

[0059] The above shows and describes the basic principles, main features and advantages of the present invention. It should be understood by those skilled in the art that the present invention is not limited to the above embodiments, and the above embodiments and descriptions are only for explaining the principles of the present invention. Without departing from the spirit and scope of the present invention, the present invention may have various changes and improvements, which fall within the scope of the present invention to be protected. The scope of protection of the present invention is defined by the attached claims and their equivalents.

Claims

1. Application of YE6144 in the preparation of drugs for the treatment of skeletal muscle atrophy caused by breast cancer.

2. The use of YE6144 according to claim 1 in the preparation of a drug for treating skeletal muscle atrophy caused by breast cancer, characterized in that: The medicine also includes pharmaceutically acceptable excipients.

3. The use of YE6144 according to claim 1 in the preparation of a drug for treating skeletal muscle atrophy caused by breast cancer, characterized in that: The medicine is in the form of tablets, capsules, granules, powders, pills or injections.

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