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The identification of novel fragment leads by Biophysical methods for the inhibition of TbrPDEB1 as an anti-parasitic approach to Human African Trypanosomiasis

Huleani, Sergiu (2018) The identification of novel fragment leads by Biophysical methods for the inhibition of TbrPDEB1 as an anti-parasitic approach to Human African Trypanosomiasis. Master of Science by Research (MScRes) thesis, University of Kent,. (KAR id:71666)

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Abstract

This drug design research project is based on identifying novel compound 'hits' against an established drug target by various screening methods such as NMR, X-Ray Crystallography and Biochemical Assays. Trypanosoma brucei cyclic nucleotide phosphodiesterase B1 (TbrPDEB1) is a validated drug target reported to be essential in the life cycle of the Trypanosoma brucei parasite - the causative agent of Human African Trypanosomiasis, one of several Neglected Parasitic Diseases which affects underdeveloped countries, and in some cases even First World countries. Otherwise known as Sleeping Sickness, the insect-borne parasitic disease occurs in 36 sub-Saharan African countries. The estimated mortality rate is up to 500,000 deaths per year and up to 80 million people are at risk of infection. With current methods of treatment being limited, ineffective and in some cases unsafe, there is an urgent need for new, more effective and safer medication.

In this research project, the TbrPDEB1 catalytic domain construct plasmid (provided by the host laboratory at the University of Kent) was used to transform an established E. coli bacteria strain for the purpose of protein expression. A small fragment library put together by IOTA Pharmaceuticals consisting of 31 active human PDE inhibitors was initially validated by Proton NMR and used to soak TbrPDEB1 protein crystals. Ligand uptake was determined by X-Ray diffraction at the Diamond Light Source. Through the use of the three previously mentioned screening approaches, fragment binders to the TbrPDEB1 target enzyme were identified. Additionally, inhibition of the fragment binders to the target was also measured and assessed in comparison to NPD-008, a known TbrPDEB1 inhibitor of moderate potency. Out of the 31 fragments, one fragment was identified as an optimal binder and low potency inhibitor to TbrPDEB1, which underwent a similarity search follow-up thus to suggest similar chemotypes of interest for future work.

Item Type: Thesis (Master of Science by Research (MScRes))
Thesis advisor: Brown, David
Uncontrolled keywords: biochemistry biology phosphodiesterase PDE trypanosoma brucie Human african trypanosomiasis crystallography xray
Divisions: Faculties > Sciences > School of Biosciences
SWORD Depositor: System Moodle
Depositing User: System Moodle
Date Deposited: 17 Jan 2019 13:11 UTC
Last Modified: 30 May 2019 08:44 UTC
Resource URI: https://kar.kent.ac.uk/id/eprint/71666 (The current URI for this page, for reference purposes)
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