Antituberculous small-molecule compound targeting at bacterium RNA (ribonucleic acid) polymerase
A compound, technology for Mycobacterium tuberculosis, used in antibacterial drugs, organic active ingredients, resistance to vector-borne diseases, etc.
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Embodiment 1
[0015] In vitro inhibition experiment of compound RNAP-001 on bacterial RNA polymerase. For the compound RNAP-001 molecule, by detecting the reduction of the substrate ATP—Kool NC-45 RNAP Activity & Inhibitor Screening Kit (Epicentre Company): 2 μg of Escherichia coli, Staphylococcus aureus, and mycobacterium-binding RNA polymerase were mixed with The gradient diluted pyridoindole derivatives were reacted at 25°C for 20 minutes, and then 1002 μM ATP (total reaction volume 50 μL) was added. After acting at 25°C for 20 minutes, the reaction mixture was added to a 96-well microtiter plate, 50 μL per well, and Kool 50 μL of NC-45Reagent per well was allowed to stand at room temperature for 10 minutes, and the chemiluminescence value was read to indicate the amount of ATP remaining in the reaction. In the experiment, the group without compound was set as the control group. Finally, calculate the inhibitory rate of compound RNAP-001 on the activity of the target protein:
[0016] ...
Embodiment 2
[0022] In vitro inhibition experiment of compound RNAP-001 on the growth of standard bacteria. Use the standard tube dilution method recommended by the Clinical and Laboratory Standards Institute (CLSI):
[0023] 1. Inoculate the bacteria in fresh MH liquid medium and culture overnight at 37°C;
[0024] 2. Correct the bacterial solution to the 0.5 McFarland turbidity standard with fresh MH liquid medium, then dilute it with MH liquid medium at 1:200, add 1 mL to each test tube, and add 1 mL of N-( (4-fluorobenzyloxy)methyl)-O-methyl-N-(6-(trifluoromethyl)-2,3,4,9-tetrahydropyridoindole)hydroxylamine and other 6 compounds (The final concentration of solvent DMSO is maintained at 1%), cultured at 37°C for 18 hours, with 1% DMSO + bacteria as the control, and sterile medium as the blank control;
[0025] 3. Take out the tube with the lowest concentration of no bacteria growth compared with the blank control, which is the minimum inhibitory concentration of the compound RNAP-001...
Embodiment 3
[0030] In vitro inhibition experiment of compound RNAP-001 on drug-resistant Mycobacterium tuberculosis. Using the experimental method described in Example 3, the compound RNAP-001 was used to carry out antibacterial experiments on three strains of drug-resistant Mycobacterium tuberculosis collected clinically. At the same time, the commonly used clinical anti-binding drugs streptomycin (SM), ethambutol (EMB), kanamycin (KM), isoniazid (INH), rifampicin (RFP), levofloxacin (LVFX), Ofloxacin (OFLX), moxifloxacin (MOX), capreomycin (CPM), and amikacin (AMK) were used as controls.
[0031] The results showed that the compound RNAP-001 had a significant inhibitory effect on the growth of drug-resistant Mycobacterium tuberculosis collected clinically, and its MIC9 0 The values are shown in Table 5.
[0032] Table 5 MIC of clinical drug-resistant strains of Mycobacterium tuberculosis to various drugs and compound RNAP-001 90 value
[0033]
[0034] Note: The data in the dru...
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