ABSTRACT:
The
Mycobacterium abscessus
drug development pipeline is poorly populated, with particularly few validated target-lead couples to initiate
de novo
drug discovery. Trimethoprim, an inhibitor of dihydrofolate reductase (DHFR) used for the treatment of a range of bacterial infections, is not active against
M. abscessus
. Thus, evidence that
M. abscessus
DHFR is vulnerable to pharmacological intervention with a small molecule inhibitor is lacking. Here, we show that the pyrrolo-quinazoline PQD-1, previously identified as a DHFR inhibitor active against
Mycobacterium tuberculosis
, exerts whole cell activity against
M. abscessus
. Enzyme inhibition studies showed that PQD-1, in contrast to trimethoprim, is a potent inhibitor of
M. abscessus
DHFR and over-expression of DHFR causes resistance to PQD-1, providing biochemical and genetic evidence that DHFR is a vulnerable target and mediates PQD-1’s growth inhibitory activity in
M. abscessus
. As observed in
M. tuberculosis
, PQD-1 resistant mutations mapped to the folate pathway enzyme thymidylate synthase (TYMS) ThyA. Like trimethoprim in other bacteria, PQD-1 synergizes with the dihydropteroate synthase (DHPS) inhibitor sulfamethoxazole (SMX), offering an opportunity to exploit the successful dual inhibition of the folate pathway and develop similarly potent combinations against
M. abscessus
. PQD-1 is active against subspecies of
M. abscessus
and a panel of clinical isolates, providing epidemiological validation of the target-lead couple. Leveraging a series of PQD-1 analogs, we have demonstrated a dynamic structure-activity relationship (SAR). Collectively, the results identify
M. abscessus
DHFR as an attractive target and PQD-1 as a chemical starting point for the discovery of novel drugs and drug combinations that target the folate pathway in
M. abscessus
.