(E/Z)-(4-(3-(2-((4-chlorophenyl)amino)-4-(dimethylamino)thiazol-5-yl)-2-(ethoxy carbonyl)-3-oxoprop-1-en-1-yl)phenyl) boronic acid, a newly developed molecule having anticancer activity serves as a potential candidate for the further drug development process. In this study, to ascertain the anticancer potential of the molecule, we screened it against different cell lines and compared the activity against the standard drug doxorubicin. The molecule showed promising activity at a low concentration against almost all cell lines used in the study. Apart from that, the molecule was characterized for its pKa and a precise reverse phase high-performance liquid chromatography bioanalytical method has been developed. The method was validated according to the United States of Food and Drug Administration bioanalytical guideline and was found to produce linear response over the calibration range of 0.8–25 μg/mL. Inter- and intra-day accuracy were found to be in the range of 93.44–99.74%, whereas precision [% coefficient of variation (CV)] for inter- and intra-day was ranged between 1.63 and 5.79%, and 0.87 and 6.96%, respectively. The bioanalytical stability testing was carried out in different conditions including 8 h benchtop, 12 h autosampler and three freeze–thaw cycles. The analyte was stable in all the tested stability conditions. Finally, an in vitro metabolite identification study was conducted using quadrupole-time of flight-mass spectrometer, and two metabolites have been identified.

中文翻译：

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生物分析和四极杆飞行时间质谱分析法驱动的基于硼酸的新型抗癌分子的体外代谢产物分析。

（E / Z）-（4-（3-（2-（（（4-氯苯基）氨基）-4-（二甲基氨基）噻唑-5-基）-2-（乙氧基羰基）-3-氧代丙-1-烯-1-基）苯基）硼酸是一种具有抗癌活性的新开发分子，可作为进一步药物开发过程的潜在候选物。在这项研究中，为了确定该分子的抗癌潜力，我们针对不同的细胞系进行了筛选，并比较了其对标准药物阿霉素的活性。该分子在低浓度下对研究中使用的几乎所有细胞系均显示出有希望的活性。除此之外，还对该分子的pKa进行了表征，并开发了一种精确的反相高效液相色谱生物分析方法。该方法已根据美国食品药品管理局的生物分析指南进行了验证，并发现在0.8–25μg/ mL的校准范围内可产生线性响应。日间和日间准确性在93.44–99.74％的范围内，而日间和日间的精度[变异系数（CV）％]介于1.63和5.79％之间，以及0.87和6.96之间％， 分别。生物分析稳定性测试是在不同的条件下进行的，包括8 h台式，12 h自动进样器和3个冻融循环。分析物在所有测试的稳定性条件下均稳定。最后，而日间和日内的精度[变异系数百分比（CV）]分别在1.63和5.79％之间，以及0.87和6.96％之间。生物分析稳定性测试是在不同的条件下进行的，包括8 h台式，12 h自动进样器和3个冻融循环。分析物在所有测试的稳定性条件下均稳定。最后，而日间和日内的精确度[变异系数百分比（CV）]分别在1.63和5.79％之间，以及0.87和6.96％之间。生物分析稳定性测试是在不同的条件下进行的，包括8 h台式，12 h自动进样器和3个冻融循环。分析物在所有测试的稳定性条件下均稳定。最后，使用四极杆飞行时间质谱仪进行了体外代谢物鉴定研究，已鉴定出两种代谢物。