Novel Saccharomyces cerevisiae screen identifies WR99210 analogues that inhibit Mycobacterium tuberculosis dihydrofolate reductase.
The ongoing selection of multidrug-resistant strains of Mycobacterium tuberculosis has markedly reduced the effectiveness of the standard treatment regimens. Thus, there is an urgent need for new drugs that are potent inhibitors of M. tuberculosis, that exhibit favorable resistance profiles, and that are well tolerated by patients. One promising drug target for treatment of mycobacterial infections is dihydrofolate reductase (DHFR; EC 1.5.1.3), a key enzyme in folate utilization. DHFR is an important drug target in many pathogens, but it has not been exploited in the search for drugs effective against M. tuberculosis. The triazine DHFR inhibitor WR99210 has been shown to be effective against other mycobacteria. We show here that WR99210 is also a potent inhibitor of M. tuberculosis and Mycobacterium bovis BCG growth in vitro and that resistance to WR99210 occurred less frequently than resistance to either rifampin or isoniazid. Screening of drugs with M. tuberculosis cultures is slow and requires biosafety level 3 facilities and procedures. We have developed an alternative strategy: initial screening in an engineered strain of the budding yeast Saccharomyces cerevisiae that is dependent on the M. tuberculosis DHFR for its growth. Using this system, we have screened 19 compounds related to WR99210 and found that 7 of these related compounds are also potent inhibitors of the M. tuberculosis DHFR. These studies suggest that compounds of this class are excellent potential leads for further development of drugs effective against M. tuberculosis.
Gerum
A'Lissa B
AB
Department of Genome Sciences, University of Washington, Seattle, Washington 98195-7730, USA.
Ulmer
Jonathan E
JE
Jacobus
David P
DP
Jensen
Norman P
NP
Sherman
David R
DR
Sibley
Carol Hopkins
CH
D000904:Antibiotics, Antitubercular / Q000494:pharmacology*; D000995:Antitubercular Agents / Q000494:pharmacology; D004353:Drug Evaluation, Preclinical; D024881:Drug Resistance, Bacterial; D005493:Folic Acid Antagonists / Q000494:pharmacology*; D005816:Genetic Complementation Test; D007538:Isoniazid / Q000494:pharmacology; D008826:Microbial Sensitivity Tests; D009169:Mycobacterium tuberculosis / Q000187:drug effects* / Q000201:enzymology* / Q000235:genetics; D012293:Rifampin / Q000494:pharmacology; D012441:Saccharomyces cerevisiae / Q000187:drug effects* / Q000235:genetics; D013762:Tetrahydrofolate Dehydrogenase / Q000235:genetics / Q000378:metabolism*; D014227:Triazines / Q000494:pharmacology*
D000904:Antibiotics, Antitubercular; D000995:Antitubercular Agents; D005493:Folic Acid Antagonists; D014227:Triazines; C006201:BRL 6231; D013762:Tetrahydrofolate Dehydrogenase; D007538:Isoniazid; D012293:Rifampin