Intracellular Accumulation of Novel and Clinically Used TB Drugs Potentiates Intracellular Synergy

• Published in: Microbiology spectrum 2021-10, Vol.9 (2), p.e0043421
• Main Authors: Tanner, Lloyd, Mashabela, Gabriel T, Omollo, Charles C, de Wet, Timothy J, Parkinson, Christopher J, Warner, Digby F, Haynes, Richard K, Wiesner, Lubbe
• Format: Article
• Language: English
• Subjects: Animals; Antimicrobial Chemotherapy; Antitubercular Agents - chemistry; Antitubercular Agents - metabolism; Antitubercular Agents - pharmacology; artemisinin; Cell Survival - drug effects; Drug Synergism; Humans; intracellular accumulation; intracellular efficacy; macrophages; Macrophages - drug effects; Macrophages - metabolism; Macrophages - microbiology; Mice; Moxifloxacin - chemistry; Moxifloxacin - metabolism; Moxifloxacin - pharmacology; Mycobacterium tuberculosis - drug effects; phenoxazine; Research Article; Rifampin - chemistry; Rifampin - metabolism; Rifampin - pharmacology; TB chemotherapy; Tuberculosis - drug therapy; Tuberculosis - metabolism; Tuberculosis - microbiology; Tuberculosis - physiopathology
• ISSN: 2165-0497; 2165-0497
• DOI: 10.1128/Spectrum.00434-21

The therapeutic repertoire for tuberculosis (TB) remains limited despite the existence of many TB drugs that are highly active in models and possess clinical utility. Underlying the lack of efficacy is the inability of TB drugs to penetrate microenvironments inhabited by the causative agent, Mycobacterium tuberculosis, including host alveolar macrophages. Here, we determined the ability of the phenoxazine PhX1 previously shown to be active against M. tuberculosis to differentially penetrate murine compartments, including plasma, epithelial lining fluid, and isolated epithelial lining fluid cells. We also investigated the extent of permeation into uninfected and M. tuberculosis-infected human macrophage-like Tamm-Horsfall protein 1 (THP-1) cells directly and by comparing to results obtained in synergy assays. Our data indicate that PhX1 (4,750 ± 127.2 ng/ml) penetrates more effectively into THP-1 cells than do the clinically used anti-TB agents, rifampin (3,050 ± 62.9 ng/ml), moxifloxacin (3,374 ± 48.7 ng/ml), bedaquiline (4,410 ± 190.9 ng/ml), and linezolid (770 ± 14.1 ng/ml). Compound efficacy in infected cells correlated with intracellular accumulation, reinforcing the perceived importance of intracellular penetration as a key drug property. Moreover, we detected synergies deriving from redox-stimulatory combinations of PhX1 or clofazimine with the novel prenylated amino-artemisinin WHN296. Finally, we used compound synergies to elucidate the relationship between compound intracellular accumulation and efficacy, with PhX1/WHN296 synergy levels shown to predict drug efficacy. Collectively, our data support the utility of the applied assays in identifying active compounds with the potential for clinical development. This study addresses the development of novel therapeutic compounds for the eventual treatment of drug-resistant tuberculosis. Tuberculosis continues to progress, with cases of Mycobacterium tuberculosis (M. tuberculosis) resistance to first-line medications increasing. We assess new combinations of drugs with both oxidant and redox properties coupled with a third partner drug, with the focus here being on the potentiation of M. tuberculosis-active combinations of compounds in the intracellular macrophage environment. Thus, we determined the ability of the phenoxazine PhX1, previously shown to be active against M. tuberculosis , to differentially penetrate murine compartments, including plasma, epithelial lining fluid, and isolated epithelial l