A High-Throughput Screen To Identify Inhibitors of ATP Homeostasis in Non-replicating <i>Mycobacterium tuberculosis</i>

Growing evidence suggests that the presence of a subpopulation of hypoxic non-replicating, phenotypically drug-tolerant mycobacteria is responsible for the prolonged duration of tuberculosis treatment. The discovery of new antitubercular agents active against this subpopulation may help in developing new strategies to shorten the time of tuberculosis therapy. Recently, the maintenance of a low level of bacterial respiration was shown to be a point of metabolic vulnerability in <i>Mycobacterium tuberculosis</i>. Here, we describe the development of a hypoxic model to identify compounds targeting mycobacterial respiratory functions and ATP homeostasis in whole mycobacteria. The model was adapted to 1,536-well plate format and successfully used to screen over 600,000 compounds. Approximately 800 compounds were confirmed to reduce intracellular ATP levels in a dose-dependent manner in <i>Mycobacterium bovis</i> BCG. One hundred and forty non-cytotoxic compounds with activity against hypoxic non-replicating <i>M. tuberculosis</i> were further validated. The resulting collection of compounds that disrupt ATP homeostasis in <i>M. tuberculosis</i> represents a valuable resource to decipher the biology of persistent mycobacteria

A High-Throughput Screen To Identify Inhibitors of ATP Homeostasis in Non-replicating <i>Mycobacterium tuberculosis</i>

Growing evidence suggests that the presence of a subpopulation of hypoxic non-replicating, phenotypically drug-tolerant mycobacteria is responsible for the prolonged duration of tuberculosis treatment. The discovery of new antitubercular agents active against this subpopulation may help in developing new strategies to shorten the time of tuberculosis therapy. Recently, the maintenance of a low level of bacterial respiration was shown to be a point of metabolic vulnerability in <i>Mycobacterium tuberculosis</i>. Here, we describe the development of a hypoxic model to identify compounds targeting mycobacterial respiratory functions and ATP homeostasis in whole mycobacteria. The model was adapted to 1,536-well plate format and successfully used to screen over 600,000 compounds. Approximately 800 compounds were confirmed to reduce intracellular ATP levels in a dose-dependent manner in <i>Mycobacterium bovis</i> BCG. One hundred and forty non-cytotoxic compounds with activity against hypoxic non-replicating <i>M. tuberculosis</i> were further validated. The resulting collection of compounds that disrupt ATP homeostasis in <i>M. tuberculosis</i> represents a valuable resource to decipher the biology of persistent mycobacteria

A High-Throughput Screen To Identify Inhibitors of ATP Homeostasis in Non-replicating <i>Mycobacterium tuberculosis</i>

Growing evidence suggests that the presence of a subpopulation of hypoxic non-replicating, phenotypically drug-tolerant mycobacteria is responsible for the prolonged duration of tuberculosis treatment. The discovery of new antitubercular agents active against this subpopulation may help in developing new strategies to shorten the time of tuberculosis therapy. Recently, the maintenance of a low level of bacterial respiration was shown to be a point of metabolic vulnerability in <i>Mycobacterium tuberculosis</i>. Here, we describe the development of a hypoxic model to identify compounds targeting mycobacterial respiratory functions and ATP homeostasis in whole mycobacteria. The model was adapted to 1,536-well plate format and successfully used to screen over 600,000 compounds. Approximately 800 compounds were confirmed to reduce intracellular ATP levels in a dose-dependent manner in <i>Mycobacterium bovis</i> BCG. One hundred and forty non-cytotoxic compounds with activity against hypoxic non-replicating <i>M. tuberculosis</i> were further validated. The resulting collection of compounds that disrupt ATP homeostasis in <i>M. tuberculosis</i> represents a valuable resource to decipher the biology of persistent mycobacteria

Discovery of Tetrahydropyrazolopyrimidine Carboxamide Derivatives As Potent and Orally Active Antitubercular Agents

Tetrahydropyrazolo­[1,5-<i>a</i>]­pyrimidine scaffold was identified as a hit series from a <i>Mycobacterium tuberculosis</i> (Mtb) whole cell high through-put screening (HTS) campaign. A series of derivatives of this class were synthesized to evaluate their structure–activity relationship (SAR) and structure–property relationship (SPR). Compound <b>9</b> had a promising in vivo DMPK profile in mouse and exhibited potent in vivo activity in a mouse efficacy model, achieving a reduction of 3.5 log CFU of Mtb after oral administration to infected mice once a day at 100 mg/kg for 28 days. Thus, compound <b>9</b> is a potential candidate for inclusion in combination therapies for both drug-sensitive and drug-resistant TB