In vitro potency of 2-(((2-hydroxyphenyl)amino)methylene)-5,5-dimethylcyclohexane-1,3-dione against drug-resistant and non-replicating persisters of Mycobacterium tuberculosis

• Main Authors: Muzafar Ahmad Rather, Zubair Shanib Bhat, Ali Mohd Lone, Mubashir Maqbool, Bilal Ahmad Bhat, Zahoor Ahmad
• Format: Article
• Language: English
• Published: Elsevier 2021-06-01
• Series: Journal of Global Antimicrobial Resistance
• Subjects: PAMCHD; Multidrug-resistant TB; Mycobacterium tuberculosis; Resistance; Tolerance; Persistence
• Online Access: http://www.sciencedirect.com/science/article/pii/S2213716521000813

ABSTRACT: Objectives: New antituberculosis agents active against drug-resistant and non-replicating tubercle bacilli are required. We evaluated a previously identified hit, 2-(((2-hydroxyphenyl)amino)methylene)-5,5-dimethylcyclohexane-1,3-dione (PAMCHD), against several clinical Mycobacterium tuberculosis isolates, including multidrug-resistant (MDR) strains and non-replicating drug-tolerant persisters of M. tuberculosis H37Rv. Methods: PAMCHD's potential against drug-resistant M. tuberculosis was investigated by broth microdilution. CFU enumeration was performed to determine PAMCHD's activity against five types of dormant bacilli. Results: No significant differences in MICs of PAMCHD were observed against M. tuberculosis H37Rv (2.5–5 µg/mL) and eight drug-susceptible strains (1.25–5 µg/mL) as well as drug-resistant strains including six isoniazid (INH)-resistant (2.5–10 µg/mL), one INH + ethambutol (EMB)-resistant (5 µg/mL), one rifampicin (RIF) + EMB-resistant (5 µg/mL) and three MDR (2.5–10 µg/mL) strains. Thus, PAMCHD maintains activity against all kinds of clinical strains, especially MDR. Regarding drug-tolerant persisters, INH and RIF killed, respectively, 0.5 and 5.0 log10 CFU of non-replicating persisters developed by hypoxia and 1.5 and 2.5 log10 CFU developed by nutrient starvation at 64 × of their respective MIC against actively dividing cultures. In contrast, PAMCHD sterilised persister cultures developed by hypoxia (killed 6.5 log10 CFU) or starvation (killed 7.5 log10 CFU). PAMCHD sterilised RIF-tolerant (tolerance level up to 100 µg/mL of RIF) 100-day-old static persisters at 64 × MIC, while moxifloxacin killed only 1.0 log10 CFU of these persisters at 64 × MIC. Conclusion: PAMCHD offers significant potential against MDR-TB and exhibits notable potency against non-replicating drug-tolerant M. tuberculosis persisters. These findings warrant further studies of PAMCHD for further anti-TB drug development.