Abstract
Drug therapies currently used for second stage Human African Trypanosomiasis (HAT) exhibit problems with toxicity, difficulty of administration, and resistance linked to the loss of transporter function. Key to the development of new drugs for HAT is a better understanding of the transport properties of candidate compounds. Standard methods for studying transport utilize radio-labelled permeant or HPLC-MS, however the natural fluorescence of many trypanocidal compounds can be exploited. Here we present a fluorescence-based assay for measuring uptake, by trypanosomes, of CPD0801, a drug candidate for second stage HAT. Sample fluorescence is measured in a 96-well format using a benchtop fluorimeter. Our method is directly applicable to the study of other diamidines with similar fluorescent properties and readily adapted for use with other cell types or fluorescent molecules as we demonstrate for the veterinary trypanocide ethidium.
References
Apr 1, 1990·Molecular and Biochemical Parasitology·B HammM Duszenko
Jul 1, 1982·Molecular and Biochemical Parasitology·P BorstJ James
May 1, 1995·Acta Tropica·P StevensonE K Mwangi
Jan 14, 1993·Nature·N S Carter, A H Fairlamb
Apr 1, 2000·Molecular and Biochemical Parasitology·H P de KoningS M Jarvis
Sep 12, 2001·Molecular and Biochemical Parasitology·E MatovuT Seebeck
Nov 9, 2001·Acta Tropica·H P de Koning, S M Jarvis
May 24, 2003·Trends in Parasitology·Patrick G BrayHarry P de Koning
Apr 24, 2004·Antimicrobial Agents and Chemotherapy·Harry P de KoningMichael P Barrett
Jul 26, 2005·FEMS Microbiology Reviews·Harry P de KoningRichard J S Burchmore
Aug 9, 2005·Lancet·Mhairi L StewartMichael P Barrett
May 26, 2006·Antimicrobial Agents and Chemotherapy·Amanda M MathisJames Edwin Hall
Aug 17, 2006·Molecular Pharmacology·Charlotte A LanteriMichael P Barrett
May 23, 2007·Antimicrobial Agents and Chemotherapy·Amanda M MathisJames Edwin Hall
Aug 30, 2008·Annals of Neurology·Peter G E Kennedy
Oct 25, 2008·Transactions of the Royal Society of Tropical Medicine and Hygiene·Manica BalasegaramFrancesco Checchi
Jul 22, 2009·Antimicrobial Agents and Chemotherapy·Tanja WenzlerReto Brun
Citations
Apr 9, 2013·Parasitology·M N C SoeiroA Hemphill
Sep 6, 2011·Nucleic Acids Research·Edwin A LewisW David Wilson
Mar 26, 2011·Molecular Pharmacology·Ibrahim A TekaHarry P de Koning
Mar 16, 2011·Antimicrobial Agents and Chemotherapy·Christopher P WardMichael P Barrett
Aug 16, 2016·PLoS Neglected Tropical Diseases·Anthonius A EzeHarry P De Koning
Feb 20, 2014·Antimicrobial Agents and Chemotherapy·Matthew K Gould, Achim Schnaufer