PDF(Pubmed)
Abstract：
Human sulfotransferase 1As (hSULT1As) play a crucial role in the metabolic clearance and detoxification of a diverse range of endogenous and exogenous substances, as well as in the bioactivation of some procarcinogens and promutagens. Pharmacological inhibiting hSULT1As activities may enhance the in vivo effects of most hSULT1As drug substrates and offer protective strategies against the hSULT1As-mediated bioactivation of procarcinogens. To date, a fluorescence-based high-throughput assay for the efficient screening of hSULT1As inhibitors has not yet been reported. In this work, a fluorogenic substrate (HN-241) for hSULT1As was developed through scaffold-seeking and structure-guided molecular optimization. Under physiological conditions, HN-241 could be readily sulfated by hSULT1As to form HN-241 sulfate, which emitted brightly fluorescent signals around 450 nm. HN-241 was then used for establishing a novel fluorescence-based microplate assay, which strongly facilitated the high-throughput screening of hSULT1As inhibitors. Following the screening of an in-house natural product library, several polyphenolic compounds were identified with anti-hSULT1As activity, while pectolinarigenin and hinokiflavone were identified as potent inhibitors against three hSULT1A isozymes. Collectively, a novel fluorescence-based microplate assay was developed for the high-throughput screening and characterization of hSULT1As inhibitors, which offered an efficient and facile approach for identifying potent hSULT1As inhibitors from compound libraries.
摘要：
人磺基转移酶1As（hSULT1As）在各种内源性和外源性物质的代谢清除和解毒中起着至关重要的作用，以及某些原致癌物和前诱变剂的生物活化。药理学抑制hSULT1As活性可以增强大多数hSULT1As药物底物的体内作用，并提供针对hSULT1As介导的原致癌物生物活化的保护策略。迄今为止,尚未报道基于荧光的高通量试验来有效筛选hSULT1As抑制剂。在这项工作中,通过寻求支架和结构引导的分子优化开发了用于hSULT1As的荧光底物（HN-241）。在生理条件下,HN-241可以很容易地被hSULT1As硫酸化以形成HN-241硫酸盐，在450nm左右发出明亮的荧光信号。然后将HN-241用于建立一种新型的基于荧光的微孔板测定，这极大地促进了hSULT1As抑制剂的高通量筛选。经过内部天然产品库的筛选，几种多酚化合物被鉴定具有抗hSULT1As活性，而果胶蛋白原蛋白和鼻烟黄酮被鉴定为三种hSULT1A同工酶的有效抑制剂。总的来说，开发了一种新型的基于荧光的微孔板测定法，用于高通量筛选和表征hSULT1As抑制剂，这为从化合物库中鉴定有效的hSULT1As抑制剂提供了一种有效而简便的方法。
