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N-myristoyltransferase inhibitors as new leads to treat sleeping sickness

Abstract

African sleeping sickness or human African trypanosomiasis, caused by Trypanosoma brucei spp., is responsible for 30,000 deaths each year. Available treatments for this disease are poor, with unacceptable efficacy and safety profiles, particularly in the late stage of the disease when the parasite has infected the central nervous system. Here we report the validation of a molecular target and the discovery of associated lead compounds with the potential to address this lack of suitable treatments. Inhibition of this target—T. brucei N-myristoyltransferase—leads to rapid killing of trypanosomes both in vitro and in vivo and cures trypanosomiasis in mice. These high-affinity inhibitors bind into the peptide substrate pocket of the enzyme and inhibit protein N-myristoylation in trypanosomes. The compounds identified have promising pharmaceutical properties and represent an opportunity to develop oral drugs to treat this devastating disease. Our studies validate T. brucei N-myristoyltransferase as a promising therapeutic target for human African trypanosomiasis.

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Figure 1: Identification of NMT lead series inhibitors.
Figure 2: TbNMT inhibitor cures acute trypanosomiasis in vivo.
Figure 3: TbNMT inhibitors have rapid trypanocidal effects in vitro and in vivo.
Figure 4: Pyrazole sulphonamide series acts ‘on target’ in the trypanosome.
Figure 5: Characterization of pyrazole sulphonamide interactions with NMT.

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Primary accessions

Protein Data Bank

Data deposits

Atomic coordinates and structure factors for the crystal structures have been deposited with the Protein Data Bank under accession codes 3H5Z and 2WSA for LmNMT with bound myristoyl CoA and with bound DDD85646, respectively.

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Acknowledgements

This work was supported by grants from the Wellcome Trust (WT077705, WT083481, WT077503 and WT085622), Scottish Funding Council (HR04013) and by the Translational Biology Theme of SULSA. We thank the European Regional Development Fund and the Wolfson Foundation for grants that provided relevant infrastructure for this work. The Structural Genomics Consortium is a registered charity (number 1097737) that receives funds from the Canadian Institutes for Health Research, the Canadian Foundation for Innovation and Genome Canada through the Ontario Genomics Institute, GlaxoSmithKline, Karolinska Institutet, the Knut and Alice Wallenberg Foundation, the Ontario Innovation Trust, the Ontario Ministry for Research and Innovation, Merck, the Novartis Research Foundation, the Swedish Agency for Innovation Systems, the Swedish Foundation for Strategic Research and the Wellcome Trust. We would like to thank all members of the Drug Discovery Unit for their technical assistance in this study, particularly B. Rao, I. Collie and D. James.

Author Contributions The project management team responsible for experimental design and coordination of research activities comprised S.B., R.B., A.H.F., M.A.J.F., J.A.F., I.H.G., K.D.R., D.M.F.vA., P.G.W. and D.F.S. J.A.B., M.H. and A.J.W. optimized expression and produced the active TbNMT used for screening in the Drug Discovery Unit. Biological studies were carried out by S.P.M., O.S., L.S.T., M.L.S.G., I.H. and H.P.P.; chemical syntheses by L.A.T.C. and S.B.; structural biology and modelling by D.A.R., O.G.R., C.P.M., R.H. and W.Q.; and pharmacological studies by L.S.

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Correspondence to Paul G. Wyatt.

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P.G.W. and S.B. are inventors on the patent (PCT/GB2009/002084).

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Frearson, J., Brand, S., McElroy, S. et al. N-myristoyltransferase inhibitors as new leads to treat sleeping sickness. Nature 464, 728–732 (2010). https://doi.org/10.1038/nature08893

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