New application of axitinib
A technique for obesity, disease, applied in the field of medicinal chemistry
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Embodiment 1
[0024] Example 1 Axitinib dose-dependently promotes browning of white adipocytes in vitro
[0025] Isolation of white adipose-derived mesenchymal cells in the groin
[0026] 1) Take an adult mouse about 10 weeks old, kill it by neck dislocation, soak it in 75% alcohol for 5 minutes, then transfer it to the operating table, and take out the white fat in the groin;
[0027] 2) Wash 3 times with PBS, cut up the adipose tissue with scissors, add 10ml collagenase I solution (prepared with D-Hanks solution, add 0.1g collagenase I to 100ml) for 1g adipose tissue, and place it at 37°C for 40 minutes;
[0028] 3) Filter with a 250 μm filter membrane, add cell culture medium, then transfer to a centrifuge tube, and centrifuge at 1000 rpm for 3 minutes;
[0029] 4) The supernatant layer after centrifugation is mature adipocytes, which is transferred to a centrifuge tube filled with PBS solution, mixed evenly and then centrifuged again. The upper mature adipocytes separated again can be...
Embodiment 210
[0055] Example 210-week-old mice were fed with a high-fat feed while feeding with axitinib at a dose of 10 mg / kg, and the blank group was fed with the same dose of PBS. After 8 weeks, the brown color of the white adipocytes in the groin was detected. Changes in weight and body weight of mice and internal organs.
[0056] Gene expression characterization of adipose tissue
[0057] RNA extraction: using the Trizol method to extract, the specific operation is as follows:
[0058] Weigh 100 mg of adipose tissue isolated from mice, then add 1 ml of Trizol, grind it with a mortar for 30 minutes, and then transfer it to an EP tube; the rest of the steps are the same as the identification of gene expression in adipocytes.
[0059] The result is as figure 2 , image 3 and Figure 4 As shown, compared with the control group, the expression of UCP1 in iBAT and iWAT in axitinib-administered mice was significantly increased, the body weight was significantly decreased, and the weight ...
Embodiment 310
[0060] Example 3 10-week-old mice were fed with a high-fat diet while being fed with axitinib at a dose of 10 mg / kg. After 8 weeks, the glucose tolerance and insulin sensitivity in the body were detected.
[0061] Glucose Tolerance and Insulin Tolerance Tests
[0062] Prepare 20% glucose solution with physiological saline, then inject mice according to the dose of 2g glucose / kg body weight, and intraperitoneally inject mice that have fasted for 12 hours (9 o'clock in the evening to 9 o'clock in the morning of the next day). The blood glucose concentration of mice at different time points (15 minutes, 30 minutes, 60 minutes, 120 minutes) before injection (0 minute) and after injection was monitored with a blood glucose meter, and the time error was controlled within 5 seconds.
[0063] The insulin dose used in the insulin resistance test was 0.5 unit / kg body weight. However, the fasting time is 6 hours (9 am to 3 pm), and the rest of the operation methods are consistent with t...
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