Study On Anti-tuberculosis Activity Of Quinoline Compounds Based On Drug Resistance Of Mycobacterium Tuberculosis

Tuberculosis has long been a serious threat to human health.Diarylquinoline compounds have considerable prospects in the treatment and research of tuberculosis.Therefore,it was laid a foundation for the research of anti-tuberculosis drugs and the later study of drug-resistant Mycobacterium tuberculosis to explore the sensitivity,drug resistance genes for Mycobacterium tuberculosis and it's safety.The study was that the minimum inhibitory concentration(MIC)of bedaquiline against Mycobacterium tuberculosis was determined by 2-fold dilution method,and its drug resistance genes were detected by sequencing.Whole genome sequencing was used to predict drug-resistant genes and pathways.The MIC and minimum bactericidal concentration(MBC)of diarylquinoline compounds against Mycobacterium tuberculosis were determined by proportional method to verify the effective perssad of bedaquiline,and the safety was investigated by in vivo toxicity test.The results are as follows:1.The total drug resistance and multidrug resistance rate of TB were 39.04% and 19.99% respectively in the region.Multidrug resistance was higher than other types of resistance except in the 71-80 age group,and the highest resistance was 28.74% in the 41-50 age group.The genome size of the Mycobacterium tuberculosis was 4406812 bp,the G/C occupying 65.62%,containing 4113 ORFs,and mainly below 600 Aa of the total protein length.Genes are mainly involved in energy production and conversion,amino acid transport and metabolism,carbohydrate transport and metabolism and lipid transport and metabolism.It was mainly taking part in the biosynthesis of ABC transporters,two-component systems,biosynthesis of amino acids,carbon metabolism,citric acid cycle,fatty acid biosynthesis and bacterial secretion system.The genes encode glycoside hydrolases,glycosyl transferases and carbohydrate esterases accounting for 82.45% of the carbohydrate-related enzymes.Mycobacterium tuberculosis had mutations in the genes encoding of aminoglycoside N-acetyltransferase,undecenyl pyrophosphate phosphatase and pentapeptide repeat family gene.2.The MIC of bedaquiline against clinical strains was mainly around 0.060?g/mL.Drug resistance gene Rv0678 of bedaquiline mainly exist V152A gene mutations and mutation rate was 5.8%,in addition to other mutations,such as G77 R,S51F,E112 K and L141R.The atpE gene mainly exist P62A mutations and mutation rate was 8.2%,in addition to D27G and V74L.PepQ mainly exist H211Q gene mutation and the mutation rate was 3.4%,no other mutations.Only H2 in 11 diarylquinoline compounds was sensitive to standard strains of Mycobacterium tuberculosis.The MIC and MBC of compound H2 to standard strains,resistance-Streptomycin,multidrug resistance and drug-multy of Mycobacterium tuberculosis respectively were 0.1?g/mL and 0.10?0.25?g/mL.The MIC and MBC of Isoniazid to standard strains of Mycobacterium tuberculosis were in 0.10?0.25?g/mL.The MBC of Rifampicin to standard strains of Mycobacterium tuberculosis was in 10?20?g/mL.The MIC of Ethambutol to standard strains of Mycobacterium tuberculosis was 0.5?g/mL.In vivo acute toxicity test shows that compared with control group,high dose group mice body quality changes have significant difference(P<0.05),and red blood cell count,platelet and platelet volume have difference differences(P<0.05),and liver coefficient have significant difference(P<0.01).The effects of drugs on organs of mice mainly shows that kidney tissues had mild inflammation and edema,and renal tubular epithelial had mild blisters degeneration,that the liver had mild edema and necrosis of liver cells,that heart and lungs not existed obvious pathological changes.Conclusion:1.The TB resistance has a high level.It was most likely to develop multi-resistance tuberculosis among resistance.The Mycobacterium tuberculosis gene was mainly involved in membrane material transport,central carbon metabolism,mainly encoded glycoside hydrolases,glycosyl transferases and carbohydrate esterases.Phenotypic resistance was related to genes.There are potential gene resistance sites.2.The MIC of bedaquiline was around 0.060?g/mL.The main genes mutation sites of Rv0678,atpE and PepQ were V152A,P62A and H211Q respectively.The mutation hotspots of V74L and G77R were found in atpE and Rv0678 genes.Diarylquinoline compound H2 has a good antibacterial effect on standard strains of Mycobacterium tuberculosis and drug-resistant strains,effecting on the changes of body mass and peripheral blood in mice.It has mild toxicity to the kidney and liver of mice.