Comparison of biotransformation and efficacy of aminoacetonitrile anthelmintics in vitro.
The present in vitro study was designed to test and compare anthelmintic activity, hepatotoxicity, and biotransformation of four selected aminoacetonitrile derivatives (AADs): monepantel (MOP, anthelmintic approved for the treatment), AAD-970, AAD-1154, and AAD-1336. Micro-agar larval development test, MTT test of cytotoxicity, and biotransformation study coupled with Ultra high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) technique were used for this purpose. Larvae of two Haemonchus contortus strains (drug susceptible and multi-drug resistant) and primary cultures of rat and ovine hepatocytes served as model systems. All AADs (including MOP) exhibited significant larvicidal effect in H. contortus susceptible as well as multi-resistant strains, much higher than those of reference anthelmintics thiabendazole and flubendazole. AAD-1154 provides the best results for most tested parameters among all AADs in this study. The cytotoxicity test showed that all AADs can be considered as nontoxic for hepatocytes. In the biotransformation study, Phase I and Phase II metabolites of AADs were identified and schemes of possible metabolic pathways in ovine hepatocytes were proposed. Biotransformation of MOP was much more extensive than biotransformation of other AADs. Based on obtained results, AAD-1154 and AAD-1336 can be considered as promising candidates for further in vivo testing.
Department of Biochemical Sciences, Faculty of Pharmacy in Hradec Králové, Charles University in Prague, Heyrovského 1203, 500 05, Hradec Králové, Czech Republic.
Lucie
L
Stuchlíková
Department of Biochemical Sciences, Faculty of Pharmacy in Hradec Králové, Charles University in Prague, Heyrovského 1203, 500 05, Hradec Králové, Czech Republic.
Lenka
L
Lecová
Department of Analytical Chemistry, Faculty of Chemical Technology, University of Pardubice, Studentská 573, 532 10, Pardubice, Czech Republic.
Robert
R
Jirásko
Department of Pharmacology and Toxicology, Faculty of Pharmacy in Hradec Králové, Charles University in Prague, Heyrovského 1203, 500 05, Hradec Králové, Czech Republic.
Jiří
J
Lamka
Department of Pharmacology and Toxicology, Faculty of Pharmacy in Hradec Králové, Charles University in Prague, Heyrovského 1203, 500 05, Hradec Králové, Czech Republic.
Ivan
I
Vokřál
Department of Biochemical Sciences, Faculty of Pharmacy in Hradec Králové, Charles University in Prague, Heyrovského 1203, 500 05, Hradec Králové, Czech Republic.
Barbora
B
Szotáková
Department of Analytical Chemistry, Faculty of Chemical Technology, University of Pardubice, Studentská 573, 532 10, Pardubice, Czech Republic.
Michal
M
Holčapek
Department of Biochemical Sciences, Faculty of Pharmacy in Hradec Králové, Charles University in Prague, Heyrovského 1203, 500 05, Hradec Králové, Czech Republic.
Lenka
L
Skálová
D000607:Aminoacetonitrile / Q000031:analogs & derivatives / Q000032:analysis / Q000493:pharmacokinetics* / Q000633:toxicity; D000818:Animals; D000871:Anthelmintics / Q000032:analysis / Q000493:pharmacokinetics* / Q000633:toxicity; D001711:Biotransformation; D002478:Cells, Cultured; D002851:Chromatography, High Pressure Liquid; D006189:Haemonchus / Q000187:drug effects; D022781:Hepatocytes / Q000378:metabolism; D007814:Larva; D008463:Mebendazole / Q000031:analogs & derivatives / Q000032:analysis / Q000493:pharmacokinetics; D051381:Rats; D017208:Rats, Wistar; D012756:Sheep; D053719:Tandem Mass Spectrometry; D013827:Thiabendazole / Q000032:analysis / Q000493:pharmacokinetics
D000871:Anthelmintics; D000607:Aminoacetonitrile; D008463:Mebendazole; C533978:monepantel; D013827:Thiabendazole; C018945:flubendazole
drug metabolism; hepatocytes; micro-agar larval development test; monepantel; structure-metabolism relationships