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ACS Infect Dis. 2017 Jul 14;3(7):512-526. doi: 10.1021/acsinfecdis.7b00022. Epub 2017 Apr 7.

Identification of Trypanosoma brucei AdoMetDC Inhibitors Using a High-Throughput Mass Spectrometry-Based Assay.

Author information

1
Genzyme Corp. (now Sanofi Genzyme) , 153 Second Avenue, Waltham, Massachusetts 02451-1122, United States.
2
Scynexis, Inc. (now Avista Pharma Solutions) , 3501 Tricenter Boulevard, Suite C, Durham, North Carolina 27713, United States.

Abstract

Human African trypanosomiasis (HAT) is a fatal infectious disease caused by the eukaryotic pathogen Trypanosoma brucei (Tb). Available treatments are difficult to administer and have significant safety issues. S-Adenosylmethionine decarboxylase (AdoMetDC) is an essential enzyme in the parasite polyamine biosynthetic pathway. Previous attempts to develop TbAdoMetDC inhibitors into anti-HAT therapies failed due to poor brain exposure. Here, we describe a large screening campaign of two small-molecule libraries (∼400,000 compounds) employing a new high-throughput (∼7 s per sample) mass spectrometry-based assay for AdoMetDC activity. As a result of primary screening, followed by hit confirmation and validation, we identified 13 new classes of reversible TbAdoMetDC inhibitors with low-micromolar potency (IC50) against both TbAdoMetDC and T. brucei parasite cells. The majority of these compounds were >10-fold selective against the human enzyme. Importantly, compounds from four classes demonstrated high propensity to cross the blood-brain barrier in a cell monolayer assay. Biochemical analysis demonstrated that compounds from eight classes inhibited intracellular TbAdoMetDC in the parasite, although evidence for a secondary off-target component was also present. The discovery of several new TbAdoMetDC inhibitor chemotypes provides new hits for lead optimization programs aimed to deliver a novel treatment for HAT.

KEYWORDS:

AdoMetDC; Trypanosoma brucei; high-throughput screening; human African trypanosomiasis; mass spectrometry

PMID:
28350440
PMCID:
PMC5511061
DOI:
10.1021/acsinfecdis.7b00022
[Indexed for MEDLINE]
Free PMC Article

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