Development of Small-Molecule Cryptochrome Stabilizer Derivatives as Modulators of the Circadian Clock.
Development of Small-Molecule Cryptochrome Stabilizer Derivatives as Modulators of the Circadian Clock.
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DOI：
10.1002/cmdc.201500260
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发表时间：
2015-09
期刊：
影响因子：
3.4
通讯作者：
Kay SA
中科院分区：
文献类型：
--
作者：
Lee JW;Hirota T;Kumar A;Kim NJ;Irle S;Kay SA
小分子探针在加深我们对生物钟的分子基础的理解方面一直发挥着重要的作用。我们之前发现了咔唑类化合物KL001(N-(3-(9H-carbazol-9-yl)-2-hydroxypropyl)-N-(furan-2-ylmethyl)methanesulfonamide)，作为时钟蛋白隐色素(CRY)的稳定剂。在这里，我们描述了KL001衍生物的广泛的构效关系分析，导致开发出一种高活性的衍生物：2-(9H-carbazol-9-yl)-N-(2-chloro-6-cyanophenyl)acetamide(KL044)。随后的3D-QSAR分析确定了KL001衍生物的关键特征，并在分子水平上了解了它们与CRY的相互作用。富含电子的咔唑、酰胺/羟基连接物、磺酰基和吸电子的丁腈基团有助于提高生物活性。与丝氨酸394和组氨酸357的氢键作用以及与Trp290的更强的CH-π相互作用使KL044成为比KL001更好的粘结剂。KL044延长了昼夜节律周期，抑制了PER2活性，并在报告分析中稳定了哭声，效价大约是KL001的10倍。总之，KL044是一种强大的化学工具，可以通过对CREY的作用来控制生物钟的功能。
Small-molecule probes have been playing prominent roles in furthering our understanding of the molecular underpinnings of the circadian clock. We previously discovered a carbazole derivative, KL001 (N-(3-(9H-carbazol-9-yl)-2-hydroxypropyl)-N-(furan-2-ylmethyl)methanesulfonamide), as a stabilizer of the clock protein cryptochrome (CRY). Herein we describe an extensive structure–activity relationship analysis of KL001 derivatives leading to the development of a highly active derivative: 2-(9H-carbazol-9-yl)-N-(2-chloro-6-cyanophenyl)acetamide (KL044). Subsequent 3D-QSAR analysis identified critical features of KL001 derivatives and provided a molecular-level understanding of their interaction with CRY. The electron-rich carbazole, amide/hydroxy linker, sulfonyl group, and electron-withdrawing nitrile moieties contribute to greater biological activity. The hydrogen bonding interactions with Ser394 and His357 as well as stronger CH–π interactions with Trp290 make KL044 a better binder than KL001. KL044 lengthened the circadian period, repressed Per2 activity, and stabilized CRY in reporter assays with roughly tenfold higher potency than KL001. Altogether, KL044 is a powerful chemical tool to control the function of the circadian clock through its action on CRY.