Screening the MayBridge Rule of 3 Fragment Library for Compounds That Interact with the Trypanosoma brucei myo-Inositol-3-Phosphate Synthase andor Show Trypanocidal Activity

Abstract EN

Inositol-3-phosphate synthase (INO1) has previously been genetically validated as a drug target against Trypanosoma brucei, the causative agent of African sleeping sickness.

Chemical intervention of this essential enzyme could lead to new therapeutic agents.

Unfortunately, no potent inhibitors of INO1 from any organism have been reported, so a screen for potential novel inhibitors of T.

brucei INO1was undertaken.

Detection of inhibition of T.

brucei INO1 is problematic due to the nature of the reaction.

Direct detection requires differentiation between glucose-6-phosphate and inositol-3-phosphate.

Coupled enzyme assays could give false positives as potentially they could inhibit the coupling enzyme.

Thus, an alternative approach of differential scanning fluorimetry to identify compounds that interact with T.

brucei INO1 was employed to screen ~670 compounds from the MayBridge Rule of 3 Fragment Library.

This approach identified 38 compounds, which significantly altered the Tm of TbINO1.

Four compounds showed trypanocidal activity with ED50s in the tens of micromolar range, with 2 having a selectivity index in excess of 250.

The trypanocidal and general cytotoxicity activities of all of the compounds in the library are also reported, with the best having ED50S of ~20 μM against T.

brucei.