title: Application of monoclonal antibodies to measure metabolism of an anti-trypanosomal compound in vitro and in vivo.
abstract: Human African trypanosomiasis (HAT), also called African sleeping sickness, is a neglected tropical parasitic disease indigenous to sub-Saharan Africa. Diamidine compounds, including pentamidine and CPD-0801, are potent anti-trypanosomal molecules. The latter is a potential drug in the development at the UNC based Consortium for Parasitic Drug Development. An orally bioavailable prodrug of CPD-0801, DB868, is metabolized primarily in the liver to the active form. A monoclonal antibody developed against a pentamidine derivative has shown significant reactivity with CPD-0801 (EC(50) 65.1 nM), but not with the prodrug (EC(50)>18,000 nM). An inhibitory enzyme-linked immunosorbent assay (IELISA) has been used to quantitatively monitor prodrug metabolism by detecting the production of the active compound over time in a sandwich culture rat hepatocyte system and in rats. These results were compared with the results of the standard LC/MS/MS assay. Spearman coefficients of 0.96 and 0.933 (in vitro and in vivo, respectively) indicate a high correlation between these two measurement methods. This novel IELISA provides a facile, inexpensive, and accurate method for drug detection that may aide in elucidating the mechanisms of action and toxicity of existing and future diamidine compounds.
authors.affiliation: Department of Pathology and Laboratory Medicine, School of Medicine, University of North Carolina, Chapel Hill, North Carolina 27599-7525, USA.
authors.forename: Rachel Beaulieu
authors.initials: RB
authors.lastname: Goldsmith
authors.forename: Danny R
authors.initials: DR
authors.lastname: Gray
authors.forename: Zhixia
authors.initials: Z
authors.lastname: Yan
authors.forename: Claudia N
authors.initials: CN
authors.lastname: Generaux
authors.forename: Richard R
authors.initials: RR
authors.lastname: Tidwell
authors.forename: Howard M
authors.initials: HM
authors.lastname: Reisner
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis / Q000276:immunology*; D000918:Antibody Specificity / Q000276:immunology; D001550:Benzamidines / Q000276:immunology; D003429:Cross Reactions / Q000276:immunology; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D005260:Female; D022781:Hepatocytes / Q000378:metabolism; D006801:Humans; D007074:Immunoglobulin G / Q000096:biosynthesis / Q000276:immunology; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D010419:Pentamidine / Q000031:analogs & derivatives / Q000276:immunology; D011355:Prodrugs / Q000378:metabolism*; D051381:Rats; D017207:Rats, Sprague-Dawley; D017208:Rats, Wistar; D053719:Tandem Mass Spectrometry; D014344:Trypanocidal Agents / Q000032:analysis* / Q000097:blood / Q000276:immunology / Q000378:metabolism*; D014353:Trypanosomiasis, African / Q000188:drug therapy
chemical_list: D000911:Antibodies, Monoclonal; D001550:Benzamidines; D007074:Immunoglobulin G; D011355:Prodrugs; D014344:Trypanocidal Agents; D010419:Pentamidine; C033286:furamidine; C458513:pafuramidine