Stuchlíková Lucie, Lecová Lenka, Jirásko Robert, Lamka Jiří, Vokřál Ivan, Szotáková Barbora, Holčapek Michal, Skálová Lenka
Department of Biochemical Sciences, Faculty of Pharmacy in Hradec Králové, Charles University in Prague, Heyrovského 1203, 500 05, Hradec Králové, Czech Republic.
Department of Analytical Chemistry, Faculty of Chemical Technology, University of Pardubice, Studentská 573, 532 10, Pardubice, Czech Republic.
Drug Test Anal. 2016 Feb;8(2):214-20. doi: 10.1002/dta.1806. Epub 2015 Apr 28.
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.
本体外研究旨在测试和比较四种选定的氨基乙腈衍生物（AADs）：莫能菌素（MOP，一种已批准用于治疗的驱虫药）、AAD - 970、AAD - 1154和AAD - 1336的驱虫活性、肝毒性和生物转化。为此，采用了微量琼脂幼虫发育试验、细胞毒性MTT试验以及结合超高效液相色谱 - 串联质谱（UHPLC - MS/MS）技术的生物转化研究。两种捻转血矛线虫菌株（药物敏感和多药耐药）的幼虫以及大鼠和绵羊肝细胞的原代培养物用作模型系统。所有AADs（包括MOP）在捻转血矛线虫敏感株和多耐药株中均表现出显著的杀幼虫作用，远高于参考驱虫药噻苯达唑和氟苯达唑。在本研究的所有AADs中，AAD - 1154在大多数测试参数上提供了最佳结果。细胞毒性试验表明，所有AADs对肝细胞均可视为无毒。在生物转化研究中，鉴定了AADs的I相和II相代谢产物，并提出了绵羊肝细胞中可能的代谢途径方案。MOP的生物转化比其他AADs更为广泛。基于所得结果，AAD - 1154和AAD - 1336可被视为有前景的进一步体内测试候选物。
