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A fluorescence-based assay for the uptake of CPD0801 (DB829) by African trypanosomes.
- Source :
-
Molecular and biochemical parasitology [Mol Biochem Parasitol] 2010 Dec; Vol. 174 (2), pp. 145-9. Date of Electronic Publication: 2010 Jul 15. - Publication Year :
- 2010
-
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.<br /> (Copyright © 2010 Elsevier B.V. All rights reserved.)
- Subjects :
- Amidines pharmacology
Animals
Chromatography, High Pressure Liquid
Ethidium metabolism
Female
Fluorescence
Fluorescent Dyes metabolism
Humans
Mass Spectrometry
Rats
Rats, Wistar
Reproducibility of Results
Sensitivity and Specificity
Trypanocidal Agents pharmacology
Trypanosoma brucei brucei cytology
Trypanosoma brucei brucei isolation & purification
Trypanosomiasis, African parasitology
Amidines metabolism
Parasitology methods
Trypanocidal Agents metabolism
Trypanosoma brucei brucei drug effects
Trypanosoma brucei brucei metabolism
Subjects
Details
- Language :
- English
- ISSN :
- 1872-9428
- Volume :
- 174
- Issue :
- 2
- Database :
- MEDLINE
- Journal :
- Molecular and biochemical parasitology
- Publication Type :
- Academic Journal
- Accession number :
- 20637807
- Full Text :
- https://doi.org/10.1016/j.molbiopara.2010.07.002