Application of compound with NMN and/or NADH structure and pharmaceutically acceptable salt of compound to preparation of mycobacterium tuberculosis inhibitor
A technology of mycobacterium tuberculosis and compounds, applied in the field of biomedicine and chemistry, can solve the problem of high toxicity
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Embodiment 1
[0037] Example 1 Inhibition experiment of NMN on Mycobacterium tuberculosis in macrophages
[0038] 1. Experimental operation:
[0039] 1.1 Obtain macrophages: THP-1 cells were mixed with 5*10 5 Cells / mL were inoculated in 96-well plates, 100 uL per well, differentiated with 100 ng / mL phorbol ester (PMA) for 24 hours, and the cells adhered to the wall to obtain macrophages derived from THP-1 cells;
[0040] 1.2 Co-incubation: Infect the macrophages derived from THP-1 cells obtained above with Mycobacterium bovis BCG 1173P2 to obtain a macrophage system infected with Mycobacterium tuberculosis; wash the macrophages infected with Mycobacterium tuberculosis The extracellular Mycobacterium bovis BCG 1173P2 of the phagocyte system was cultured by adding 1640 medium containing 10% (volume fraction) fetal bovine serum; adding 1mM, 5mM, and 10mM NMN (final concentration) respectively, and setting a blank at the same time For the control group, incubated for 48 hours;
[0041] 1.3 D...
Embodiment 2
[0043] Example 2 Inhibition experiment of NADH on Mycobacterium tuberculosis in macrophages
[0044] 1. Experimental operation:
[0045] 1.1 Obtain macrophages: THP-1 cells were mixed with 5*10 5 Cells / mL were inoculated in 96-well plates, 100 uL per well, differentiated with 100 ng / mL phorbol ester (PMA) for 24 hours, and the cells adhered to the wall to obtain macrophages derived from THP-1 cells;
[0046]1.2 Co-incubation: Infect the macrophages derived from THP-1 cells obtained above with Mycobacterium bovis BCG 1173P2 to obtain a macrophage system infected with Mycobacterium tuberculosis; wash the macrophages infected with Mycobacterium tuberculosis The extracellular Mycobacterium bovis BCG 1173P2 of the phagocyte system was cultured in 1640 medium containing 10% (volume fraction) fetal bovine serum; NADH (final concentration) of 1mM, 5mM, and 10mM was added respectively, and a blank was set at the same time For the control group, incubated for 48 hours;
[0047] 1.3 D...
Embodiment 3
[0049] Embodiment 3 NMN is to the inhibition test of Mycobacterium tuberculosis
[0050] 1. Experimental operation: adjust the concentration of Mycobacterium bovis BCG 1173P2 to an OD600 reading of 0.05, inoculate in a 96-well plate, 99 uL per well. Then add 1.0 uL of the concentration gradient dilution solution of the NMN (final concentration) of 1mM, 5mM and 10mM respectively, and incubate altogether for 48h at 37°C; Mycobacterium BCG1173P2, but did not add the test drug, but added rifampicin). And take samples at 4h and 8h to detect the GFP fluorescence intensity in the culture system, and obtain as follows: image 3 The results shown; wherein, 4 replicate holes were set at each time point.
[0051] 2. Analysis of the results: when incubated for 4 hours, each concentration of NMN had no obvious inhibitory effect compared with the control group; after incubation for 8 hours, each concentration of NMN had a significant inhibitory effect on Mycobacterium tuberculosis. Among...
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