Application of celecoxib derivative OSU-03012 in preparation of anti-tubercle bacillus drugs
A technology for celecoxib and anti-tuberculosis bacteria, applied in antibacterial drugs, drug combinations, respiratory diseases, etc., can solve the unreported OSU-03012 and other problems, and achieve good clinical application prospects
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Embodiment 1
[0027] The C57BL / 6 mouse peritoneal primary macrophages were divided into 1*10 5 Each cell / well was inoculated in a 48-well cell culture plate. After about 2 hours for the cells to adhere to the wall, the complete medium 1640 was removed, and then fresh complete medium was added to culture overnight. The next day, 1 h before infection, replace with fresh 1640 containing 10% serum without double antibody, and add AR-12 at a final concentration of 1uM in the cell culture supernatant; use DMSO as a control well, place the cells in After culturing in the incubator for 1 h, the tubercle bacillus (H37Rv) was infected at a dose of MOI=5. After 2-3 hours of infection, discard the supernatant, then culture the cells with amikacin-containing medium for 2 hours, discard the supernatant and replace with 1640 containing 10% serum without double antibody and continue at 37°C for 5 %CO 2 Cells were cultured in the incubator for 24 h. Discard the supernatant and wash the cells with PBS, th...
Embodiment 2
[0030] The C57BL / 6 mice were divided into two groups, 6 in each group, and infected with Mycobacterium tuberculosis (H37Rv) by intraperitoneal injection, the dose was 2*10 6 Bacteria / mouse, 1 week after infection, orally administered 25 mg / kg and PBS as a control, and administered continuously for 3 weeks. The mice were sacrificed by dislocation of the cervical spine, and one lobe of the lung was taken out, fixed with 4% paraformaldehyde, paraffin sectioned, and H&E staining was performed to observe the pathological damage of the lung.
[0031] see figure 2 , given AR-12 at a dose of 100-200 mg / kg, after three weeks of oral administration, the pathological damage of the lungs of mice was significantly reduced; therefore, the experimental results of this example show that AR-12 can effectively reduce the damage of the lungs of mice. Pathological damage.
Embodiment 3
[0033] Take one-third of the mouse lung tissue administered three weeks after infection in Example 2, and grind it with 1 ml of PBS containing 1% triton-100, serially dilute, and take 10 -3 、10 -4Spread 100ml of tissue suspension evenly on Middle Brook 7H10 agar culture plate containing amphotericin B, then place it in a 37°C incubator for 2-3 weeks, and count the colonies. The results are as follows: image 3 shown.
[0034] The experimental results of this example show that AR-12 significantly reduces the bacterial load of Mycobacterium tuberculosis in the lungs of mice.
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