Institute of Transformative Bio-Molecules, Nagoya University, Nagoya 464-8601, Japan.
Institute of Transformative Bio-Molecules, Nagoya University, Nagoya 464-8601, Japan; Department of Chemistry, Graduate School of Science, Nagoya University, Nagoya 464-8601, Japan.
Cell Chem Biol. 2020 Sep 17;27(9):1192-1198.e5. doi: 10.1016/j.chembiol.2020.05.008. Epub 2020 Jun 4.
Cryptochrome 1 (CRY1) and CRY2 are core regulators of the circadian clock, and the development of isoform-selective modulators is important for the elucidation of their redundant and distinct functions. Here, we report the identification and functional characterization of a small-molecule modulator of the mammalian circadian clock that selectively controls CRY1. Cell-based circadian chemical screening identified a thienopyrimidine derivative KL201 that lengthened the period of circadian rhythms in cells and tissues. Functional assays revealed stabilization of CRY1 but not CRY2 by KL201. A structure-activity relationship study of KL201 derivatives in combination with X-ray crystallography of the CRY1-KL201 complex uncovered critical sites and interactions required for CRY1 regulation. KL201 bound to CRY1 in overlap with FBXL3, a subunit of ubiquitin ligase complex, and the effect of KL201 was blunted by knockdown of FBXL3. KL201 will facilitate isoform-selective regulation of CRY1 to accelerate chronobiology research and therapeutics against clock-related diseases.
CRY1（隐花色素 1）和 CRY2 是生物钟的核心调节剂，开发同工型选择性调节剂对于阐明其冗余和独特功能非常重要。在这里，我们报告了一种哺乳动物生物钟的小分子调节剂的鉴定和功能表征，该调节剂选择性地控制 CRY1。基于细胞的生物钟化学筛选鉴定出一种噻吩并嘧啶衍生物 KL201，它延长了细胞和组织中生物钟的周期。功能测定显示 KL201 稳定了 CRY1，但不是 CRY2。KL201 衍生物的构效关系研究与 CRY1-KL201 复合物的 X 射线晶体学相结合，揭示了 CRY1 调节所需的关键位点和相互作用。KL201 与 FBXL3（泛素连接酶复合物的一个亚基）在 CRY1 上重叠结合，KL201 的作用被 FBXL3 的敲低所削弱。KL201 将促进 CRY1 的同工型选择性调节，从而加速生物钟研究和针对与时钟相关疾病的治疗。
