抗乙肝病毒新药Bay41-4109的发现毒理学研究

The discovery toxicology of Bay41-4109,a novel anti-HBV compound

目的:利用发现毒理学中的临床前先导化合物优化技术(preclinical lead optimization technolo- gies,PLOTs)对抗乙肝病毒候选新药Bay41-4109的一般毒性、遗传毒性和生殖毒性进行研究,为早期发现候选新药的毒性提供实验依据。方法:一般毒性研究中分别以MTT比色法和上下法检测Bay41-4109的体外细胞毒性和小鼠LD_(50);遗传毒性研究中分别以Ames波动试验、SOS显色试验、双核细胞微核试验检测Bay41-4109诱发鼠伤寒沙门菌基因回复突变的能力、诱发大肠杆菌的原发DNA损伤效应以及对CHL细胞的染色体断裂效应;生殖毒性研究中以大鼠胚胎中脑细胞微团培养试验来检测Bay41-4109的致畸性。结果:Bay41-4109对CHL细胞的IC_(50)为54.0μmol·L^(-1),对雌性小鼠的LD_(50)大于2 000 mg·kg^(-1)。无论有无S9活化,Bay41-4109均不引起沙门菌回复突变,也不导致DNA损伤和染色体断裂。Bay41-4109对大鼠胚胎中脑细胞亦无致畸作用。结论:早期毒性筛选结果表明:Bay41-4109未表现明显的遗传毒性和生殖毒性。

Objective ： To novel anti-HBV compound, using evaluate the general toxicity, genotoxicity preclinical lead optimization technologies and teratogenecity of Bay41-4109, a during a discovery toxicology study. Methods： In the study of general toxicity, the in vitro basic cytotoxicity to CHL cells and the acute oral toxicity in mice of Bay41-4109 were determined with MTT colorimetric assay and up and down acute toxicity experiments. The genotoxicity was determined by the Ames-fluctuation test, the SOS chromotest and the in vitro cytokinesis-block mi- cronucleus test, to detect the mutagenic activity in Salmonella typhimurium, the primary DNA damage effect in Escherichia coli, and the clastogenic effect in CHL cells, respectively. The method of the micromass culture of rat embryo midbrain cells was used for evaluating teratogenecity. Results： The IC50 of Bay41-4109 to CHL cells was 54.0μmol·L-1, the LDs0 to female mice was more than 2 000 mg·kg -1 Bay41-4109 showed no mutagenicity in Salmonella typhimurium; the primary DNA damage effect in Escherichia coli and the clastogenic effect in CHL cells were also not detected with or without the S9 metabolic system. No potential teratogenicity in vitro was found for Bay41-4109. Conclusions ： Bay41-4 109 has no obvious genotoxicity and teratogenecity, and can be considered as an anti-HBV drug candidate for further development.