Institute for Quantitative Biosciences, the University of Tokyo, 1-1-1 Yayoi, Bunkyo-ku, Tokyo 113-0032, Japan.
Nihon University School of Medicine, 30-1 Oyaguchi-kamicho, Itabashi-ku, Tokyo 173-8610, Japan.
Bioorg Med Chem. 2019 May 15;27(10):1952-1961. doi: 10.1016/j.bmc.2019.03.042. Epub 2019 Mar 26.
Selective estrogen receptor (ER) down-regulators (SERDs) are pure ER antagonists that also induce ER degradation upon binding to the receptor. Although SERDs have been developed for the treatment of ER-positive breast cancers for nearly a decade, their precise mechanism(s) of action and structure-activity relationship are still unclear. Generally, Western blotting is used to examine the effects of SERDs on ER protein levels, but the methodology is low-throughput and not quantitative. Here, we describe a quantitative, high-throughput, luciferase-based assay for the evaluation of SERDs activity. For this purpose, we established stable recombinant HEK-293 cell lines expressing ERα fused with emerald luciferase. We also designed and synthesized new diphenylmethane derivatives as candidate SERDs, and evaluated their SERDs activity using the developed system in order to examine their structure-activity relationship, taking EC as a measure of potency, and E as a measure of efficacy.
选择性雌激素受体 (ER) 降解剂 (SERD) 是纯 ER 拮抗剂，与受体结合后也会诱导 ER 降解。尽管 SERD 已开发用于治疗 ER 阳性乳腺癌近十年，但它们的确切作用机制和构效关系仍不清楚。通常，Western blot 用于检测 SERD 对 ER 蛋白水平的影响，但该方法通量低且不是定量的。在这里，我们描述了一种用于评估 SERD 活性的定量、高通量、基于荧光素酶的测定法。为此，我们建立了稳定表达 ERα与祖母绿荧光素酶融合的重组 HEK-293 细胞系。我们还设计并合成了新的二苯甲烷衍生物作为候选 SERD，并使用开发的系统评估了它们的 SERD 活性，以研究它们的构效关系，以 EC 作为效力的衡量标准，以 E 作为功效的衡量标准。
