Effect of poloxamer CRL-1072 on drug uptake and nitric-oxide-mediated killing of Mycobacterium avium by macrophages

Abstract

Mycobacterium avium–intracellulare complex (MAI) are common pathogens of opportunistic infections that are naturally resistant to most antibiotics and develop acquired resistance rapidly. An experimental drug, poloxamer CRL-1072, was found to have two unusual properties: it synergistically enhanced the activity of several antibiotics against MAI even though it had little activity as a single agent and it had greater activity against MAI in macrophage culture or in mice than in broth culture. Studies were undertaken to investigate the mechanisms of these effects. CRL-1072 was taken up by MAI and enhanced the uptake of fluorescent-labeled streptomycin and erythromycin in broth culture. The labeled antibiotics had reduced activity so the relevance for naive antibiotics must be inferred. In culture with human U937 monocytoid cells, CRL-1072 became localized in phagosomes and promoted uptake of streptomycin. Finally, CRL-1072 was found to induce production of mRNA for inducible nitric oxide synthase (iNOS) and nitric oxide (NO) by U937 cells. The antimycobacterial effect in macrophages was reversed by the iNOS inhibitor N-monomethyl l-arginine (NMMA), suggesting that CRL-1072 promotes killing of MAI by inducing NO. These effects were induced by noncytotoxic concentrations of CRL-1072. These data suggest that the antimycobacterial mechanisms of CRL-1072 include enhancing the delivery of antibiotic to targets within MAI and enhancement of the ability of macrophages to kill ingested organisms.