Molecular Insight into Mycobacterium tuberculosis Resistance to Nitrofuranyl Amides Gained through Metagenomics-like Analysis of Spontaneous Mutants

Abstract

We performed synthesis of new nitrofuranyl amides and investigated their anti-TB activity and primary genetic response of mycobacteria through whole-genome sequencing (WGS) of spontaneous resistant mutants. The in vitro activity was assessed on reference strain Mycobacterium tuberculosis H37Rv. The most active compound 11 was used for in vitro selection of spontaneous resistant mutants. The same mutations in six genes were detected in bacterial cultures grown under increased concentrations of 11 (2×, 4×, 8× MIC). The mutant positions were presented as mixed wild type and mutant alleles while increasing the concentration of the compound led to the semi-proportional and significant increase in mutant alleles. The identified genes belong to different categories and pathways. Some of them were previously reported as mediating drug resistance or drug tolerance, and counteracting oxidative and nitrosative stress, in particular: Rv0224c, fbiC, iniA, and Rv1592c. Gene-set interaction analysis revealed a certain weak interaction for gene pairs Rv1592–Rv1639c and Rv1592–Rv0224c. To conclude, this study experimentally demonstrated a multifaceted primary genetic response of M. tuberculosis to the action of nitrofurans. All three 11-treated subcultures independently presented the same six SNPs, which suggests their non-random occurrence and likely causative relationship between compound action and possible resistance mechanism.

Mycobacterium tuberculosis Lineage Distribution Using Whole-Genome Sequencing and Bedaquiline, Clofazimine, and Linezolid Phenotypic Profiles among Rifampicin-Resistant Isolates from West Java, Indonesia

“…Both of those genes, the efflux pump repressor mmpR ( Rv0678 ) and the atpE gene ( Rv1305 ), specifically the F0 domain of the ATP synthase enzyme, exhibit alterations in bedaquiline-producing mutants that are resistant to the drug [ 38 , 39 ]. The uncharacterized transporter Rv1979c and pepQ , which are both encoded as genes involved in cross-resistance between bedaquiline and clofazimine, have statistically significant co-occurrences with Rv0678 [ 40 , 41 ].…”

Evaluation of Acute and Sub-Acute Toxicity, Oxidative Stress and Molecular Docking of Two Nitrofuranyl Amides as Promising Anti-Tuberculosis Agents

“…[ 8 ]. In our recent study, we performed the synthesis of small series of new nitrofuranyl amides [ 14 ]. We investigated their anti-TB activity and primary genetic response of mycobacteria (for selected single piperazine-containing amide) through whole-genome sequencing (WGS) of spontaneous resistant mutants.…”

“…Here, we selected two nitrofurane derivatives ( DO-190 and DO-209 ), synthesized and described in our previous study ( Figure 1 ), suitable for further investigation of their pharmacology and toxicology profile since they already demonstrated high in vitro antimycobacterial activity [ 14 ].…”

Tetrahydroisoquinoline based 5-nitro-2-furoic acid derivatives: a promising new approach for anti-tubercular agents

“…14,15 These NF derivatives containing typical hydrazide/hydrazone moieties may affect other infectious diseases, especially leishmaniasis, trypanosomiasis, tuberculosis, and cancer. [16][17][18][19][20][21] Several nitrofuran compounds with anti-tubercular activity have been reported. [19][20][21] Fig.…”

“…[16][17][18][19][20][21] Several nitrofuran compounds with anti-tubercular activity have been reported. [19][20][21] Fig. 1 depicts a few of the THIQ and NF analogs employed in clinical usage.…”