telacebec q203  (Qurient Inc)

Journal: eLife

Article Title: Chalkophore-mediated respiratory oxidase flexibility controls M. tuberculosis virulence

doi: 10.7554/eLife.105794

Figure Lengend Snippet: ( A ) Heat map of transcripts encoding selected respiratory chain components determined by RNA sequencing of M. tuberculosis wild-type (WT) or Δ nrp treated with TTM or Q203. WT_GSE is the published dataset GSE159080 of M. tuberculosis H37Rv treated with Q203. Genes in the chalkophore cluster are boxed in red, genes encoding the cytochrome BD (CytBD) oxidase in blue, and genes encoding components of the bcc:aa3 supercomplex are in green. ( B ) RT-qPCR of the transcript encoding CydA in M. tuberculosis WT, Δ nrp , and complemented strain treated with varying TTM concentrations for 4 hr. Error bars represent the standard error of the mean (SEM). Statistical significance determined via two-way ANOVA with Tukey correction for multiple comparisons. *p<0.05, **p<0.01, ****p<0.0001. ( C ) RT-qPCR of the transcript encoding CydA in M. tuberculosis WT, Δ nrp , and complemented strain treated with 20 μM TTM for 24 hr. Error bars are SEM. Statistical significance determined via two-way ANOVA with Tukey correction for multiple comparisons. **p<0.01, ***p<0.001, ****p<0.0001. ( D ) Dose-dependent effect of tetrathiomolybdate (TTM) on growth of the indicated M. tuberculosis strains at 7 d post inoculation. The dotted line indicates the starting inoculum. Error bars are SEM. Statistical significance determined via two-way ANOVA with Tukey correction for multiple comparisons. *p<0.05. Figure 1—source data 1. Raw data values for .

Article Snippet: Chemical compound, drug , Telacebec (Q203) , AbMole BioScience , M5297 , .

Techniques: RNA Sequencing, Quantitative RT-PCR

Journal: eLife

Article Title: Chalkophore-mediated respiratory oxidase flexibility controls M. tuberculosis virulence

doi: 10.7554/eLife.105794

Figure Lengend Snippet: ( A ) Schematic of the terminal respiratory oxidases of M. tuberculosis . The bcc:aa 3 oxidase is a heme-copper oxidase and cytochrome BD (CytBD) is a copper-independent heme oxidase. Both transfer electrons to oxygen. Q203 is an inhibitor of bcc:aa 3 by targeting the QcrB subunit, whereas ND-011992 targets CytBD. The two oxidases are individually dispensable due to compensation by the other oxidase, but M. tuberculosis lacking both is nonviable. The model to be tested is that copper chelation deprives the bcc:aa 3 oxidase of copper and that diisonitrile chalkophores counter this copper deprivation stress. ( B ) Liquid growth assays of the indicated strains with or without 20 µM TTM treatment. OD 600 at day 10 post-inoculation displayed. Dotted line indicates starting inoculum. Error bars are SEM. Statistical significance determined via two-way ANOVA with Tukey correction for multiple comparisons. ****p<0.0001. ( C ) Bacterial survival of the indicated strains on agar media containing no addition, 1 mM BCS, or 40 μM TTM. Dotted line indicates lower limit of detection (LLOD). Error bars are SEM. Statistical significance determined via two-way ANOVA with Tukey correction for multiple comparisons. **p<0.01, *p<0.05 ( D ) The copper deprivation sensitivity of M. tuberculosis Δ nrp Δ cydAB strain can be rescued with a synthetic diisonitrile chalkophore. Liquid growth assays of Δ nrp Δ cydAB with DMSO, 20 μM TTM, or 20 μM TTM with 10 μM of the diisonitrile chalkophore pictured in panel A. Error bars are SEM. ( E ) The bcc:aa 3 oxidase is the only target of copper starvation countered by diisonitrile chalkophores. Liquid growth assays of the indicated strains treated with 10 or 20 µM TTM, or DMSO vehicle control. OD 600 at day 10 post-inoculation displayed. Dotted line indicates starting inoculum. Error bars are SEM. Statistical significance determined via two-way ANOVA with Tukey correction for multiple comparisons. ns = not significant, ****p<0.0001. ( F ) The effect of copper deprivation is masked by inhibition of QcrB subunit of bcc : aa 3 . Liquid growth assays of the indicated strains treated with Q203 (100 nM) alone or co-treated with 100 nM Q203 and 10 μM TTM. OD 600 at day 7 post-inoculation displayed. Dotted line indicates starting inoculum. Error bars are SEM. Statistical significance determined via two-way ANOVA with Tukey correction for multiple comparisons. ***p<0.001, ns = not significant. Figure 2—source data 1. Raw data for .

Article Snippet: Chemical compound, drug , Telacebec (Q203) , AbMole BioScience , M5297 , .

Techniques: Control, Inhibition

Journal: eLife

Article Title: Chalkophore-mediated respiratory oxidase flexibility controls M. tuberculosis virulence

doi: 10.7554/eLife.105794

Figure Lengend Snippet: ( A ) Methylene blue decolorization assay of oxygen consumption under copper deprivation (tetrathiomolybdate, TTM) or treatment with Q203 in wild-type (WT) or Δ nrp Δ cydAB M. tuberculosis at day 0 (d0) or day 3 (d3) of incubation. Clear vials indicate oxygen consumption by respiration. ( B ) Quantitative measurement of oxygen consumption using oxygen-sensitive optical sensors. WT M. tuberculosis treated with DMSO, ND-011992, Q203, or both ND-011992 and Q203. Oxygen measurements were taken daily. Each point represents three measurements of two biological replicates. Error bars are SEM. Statistical significance between Q203 and ND-011992 +Q203 determined via two-way ANOVA with Tukey correction for multiple comparisons. ****p<0.0001. ( C ) Same assay as in panel B with WT and Δ nrp M. tuberculosis treated with DMSO or 25 μM TTM. Error bars are SEM. ( D ) Same assay as in panel B with WT, Δ nrp Δ cydAB, or Δ nrp Δ cydAB + nrp treated with 25 μM TTM. Error bars are SEM. Statistical significance between WT and Δ nrp Δ cydAB treated with 25 μM TTM determined via two-way ANOVA with Tukey correction for multiple comparisons. ****p<0.0001. ( E ) Cellular ATP levels determined by BacTiter-Glo in the indicated strains treated with DMSO, 20 or 40 μM TTM, or 100 nM Q203. [ATP] determined by standard curve determined in growth media containing the same quantities of DMSO, TTM, or Q203. Error bars are SEM. Statistical significance determined via two-way ANOVA with Tukey correction for multiple comparisons. *p<0.05, ***p<0.001, ****p<0.0001. ( F ) Relative abundance of a CtaD-ALFA protein in M. tuberculosis of the indicated genotype treated with bathocuproinedisulfonic acid (BCS) or TTM. See for primary immunoblot data. Error bars are SEM. Statistical significance determined via two-way ANOVA with Tukey correction for multiple comparisons. ns = not significant. Figure 3—source data 1. Raw data for .

Article Snippet: Chemical compound, drug , Telacebec (Q203) , AbMole BioScience , M5297 , .

Techniques: Incubation, Western Blot

Journal: eLife

Article Title: Chalkophore-mediated respiratory oxidase flexibility controls M. tuberculosis virulence

doi: 10.7554/eLife.105794

Figure Lengend Snippet: ( A ) Methylene blue decolorization assay in WT M. tuberculosis or M. tuberculosis Δ nrp at assay start (d0) or after 3 d of incubation in a sealed tube (d3) treated with DMSO, 50 µM tetrathiomolybdate (TTM), or the combination of Q203 (400 nM) and ND-011992 (50 µM). ( B ) Copper chelation does not destabilize the CtaD protein M. tuberculosis lacking the CtaD subunit of the bcc:aa3 oxidase and complemented with a fully functional CtaD with a C-terminal ALFA epitope tag (see ) either in the wild-type background (WT) or M. tuberculosis Δ nrp (KO), which lacks diisonitrile chalkophore biosynthesis, and treated with either bathocuproinedisulfonic acid (BCS) or TTM. Full immunoblots for the ALFA tag or RpoB as a loading control are shown. Images were quantified using ImageJ software. Copper chelation synergizes with CytBD inhibition in the absence of chalkophore biosynthesis. Serial dilutions of M. tuberculosis WT,!1nrp,!1nrp +nrp strains were cultured on agar media containing TTM, ND-011992 (ND), or both. *p=0.0148 by two-way ANOVA. Figure 3—figure supplement 1—source data 1. PDF file containing original western blots for , with bands labeled. Figure 3—figure supplement 1—source data 2. Original uncropped western blot files for . Figure 3—figure supplement 1—source data 3. Bacterial counts for .

Article Snippet: Chemical compound, drug , Telacebec (Q203) , AbMole BioScience , M5297 , .

Techniques: Incubation, Functional Assay, Western Blot, Control, Software, Inhibition, Cell Culture, Labeling

Journal: The Lancet. Microbe

Article Title: Addressing urgent priorities in antibiotic development: insights from WHO 2023 antibacterial clinical pipeline analyses

doi: 10.1016/j.lanmic.2024.100992

Figure Lengend Snippet: Clinical pipeline of candidate antibiotics against Mycobacterium tuberculosis in 2023

Article Snippet: Telacebec (Q203) , Imidazopyridine amide , PO , Qurient, Infectex, TB Alliance , Criterion fulfilled , Criterion fulfilled , Criterion fulfilled , Criterion fulfilled.

Techniques: Inhibition, Infection

Journal: Nature reviews. Drug discovery

Article Title: Therapeutic developments for tuberculosis and nontuberculous mycobacterial lung disease

doi: 10.1038/s41573-024-00897-5

Figure Lengend Snippet: Selected ongoing clinical trials of new agents for TB and NTM-PD

Article Snippet: Q203 (telacebec) , Yes, cytochrome bcc complex (respiratory chain) , Phase IIa dose ranging to evaluate EBA, safety, tolerability and PK in patients with drug-susceptible TB , Qurient Co. 246 , NCT03563599.

Techniques: Clinical Proteomics, Analogues, Suspension, Saline, Activity Assay