The pharmacological potential of drugs must be evaluated to establish their potential therapeutic benefits and side effects. This evaluation includes assessment of the effects of hepatic enzymes that catalyse their metabolic activation. Previously, our research group synthesized and characterized a set of synthetic 3-alkyl pyridine alkaloid (3-APA) analogues that cause in vitro cytotoxic, genotoxic, and mutagenic effects in various human cancer cell lines. The present study aimed to evaluate these activities with the two most promising synthetic 3-APAs (3-APA 1 and 3-APA 2) against cell lines derived from breast cancer (MDA-MB-231), ovarian cancer (TOV-21 G) and lung fibroblasts (WI-26-VA4) with and without metabolic activation (S9 fraction). The cytotoxicity of the compounds was evaluated employing MTT and clonogenic assays. In addition, comet assays, γH2AX immunocytochemistry labelling assays and cytokinesis-block micronucleus tests were carried out to evaluate the potential of these compounds to induce chromosomal damage. The results obtained in the MTT assay showed that compound 3-APA 2 exhibited high selectivity index (SI) values (ranging between 21.0 and 92.6). In addition, the cytotoxicity of the compounds was clearly enhanced by metabolic activation. Moreover, both compounds were genotoxic and induced double-strand breaks in DNA and chromosomal lesions with and without S9. The cancer cell lines tested showed higher genotoxic sensitivity to the compounds than did the non-tumour cell line used as a reference. The genotoxic and mutagenic effects of the compounds were potentiated in experiments with metabolic activation. The data obtained in this study indicate that compound 3-APA 2 is more active against the human cancer cell lines tested, both with and without metabolic activation, and can therefore be considered a candidate drug to treat human ovarian and breast cancer.

必须评估药物的药理学潜力，以确定其潜在的治疗益处和副作用。该评估包括评估催化其代谢活化的肝酶的作用。此前，我们的研究小组合成并表征了一组合成的 3-烷基吡啶生物碱 (3-APA) 类似物，这些类似物在各种人类癌细胞系中引起体外细胞毒性、基因毒性和诱变作用。本研究旨在用两种最有希望的合成 3-APA（3-APA 1和3-APA 2) 对抗源自乳腺癌 (MDA-MB-231)、卵巢癌 (TOV-21 G) 和肺成纤维细胞 (WI-26-VA4) 的细胞系，有和没有代谢激活（S9 部分）。使用MTT和克隆形成试验评估化合物的细胞毒性。此外，还进行了彗星试验、γH2AX 免疫细胞化学标记试验和胞质分裂阻断微核试验，以评估这些化合物诱导染色体损伤的潜力。MTT测定结果表明，化合物3-APA 2表现出高选择性指数 (SI) 值（范围在 21.0 和 92.6 之间）。此外，代谢活化明显增强了化合物的细胞毒性。此外，两种化合物都具有遗传毒性，并在有和没有 S9 的情况下诱导 DNA 和染色体损伤的双链断裂。与用作参考的非肿瘤细胞系相比，测试的癌细胞系对化合物显示出更高的基因毒性敏感性。这些化合物的基因毒性和致突变作用在代谢活化实验中得到加强。本研究中获得的数据表明，化合物3-APA 2对所测试的人类癌细胞系具有更高的活性，无论有无代谢激活，因此可被视为治疗人类卵巢癌和乳腺癌的候选药物。