Discovery of a Positive Allosteric Modulator of Cholecystokinin Action at CCK1R in Normal and Elevated Cholesterol.
Discovery of a Positive Allosteric Modulator of Cholecystokinin Action at CCK1R in Normal and Elevated Cholesterol.
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DOI：
10.3389/fendo.2021.789957
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发表时间：
2021
影响因子：
5.2
通讯作者：
Miller LJ
中科院分区：
文献类型：
--
作者：
Harikumar KG;Coudrat T;Desai AJ;Dong M;Dengler DG;Furness SGB;Christopoulos A;Wootten D;Sergienko EA;Sexton PM;Miller LJ
预防/治疗肥胖症有用的药物可以改善健康。CCK是食欲的关键调节剂，通过1型CCK受体(CCK1R)发挥作用；然而，完全激动剂并不比节食更能刺激减肥。我们提出了一种针对该受体的替代策略，同时降低了副作用和/或毒性的可能性。具有最小内源性激动剂活性的正变构调节剂(PAM)可以增强CCK的作用，同时保持生理信号的时空特性。这可以纠正在肥胖的高胆固醇环境中观察到的异常刺激-活动耦合。我们利用高通量筛选来识别具有这种药理学特征的分子，并研究其作用基础。化合物1是一种弱的部分激动剂，具有PAM活性，可增强CCK1R的CCK作用，但不能增强CCK2R的作用，在正常和高胆固醇状态下都保持。化合物1(10µM)不具有激动剂活性，也不刺激CCK1R内化。它通过减缓结合激素的脱落率，增加其结合亲和力来增强CCK的活性。化合物1与CCK1R的计算机对接产生了看似合理的姿势。一种可放射性碘化的耐光类似物保留了化合物1的药理作用，并共价标记了CCK1R Thr211，与一个建议的姿势一致。我们的研究确定了一种新的、选择性的CCK1R PAM，它与受体结合，在正常和模拟疾病的高胆固醇环境中增强CCK-8和CCK-58的作用。这促进了化合物的发展，这些化合物以CCK对CCK1R的生理、空间和时间参与为靶点，支持其在代谢调节中的关键作用。
Drugs useful in prevention/treatment of obesity could improve health. Cholecystokinin (CCK) is a key regulator of appetite, working through the type 1 CCK receptor (CCK1R); however, full agonists have not stimulated more weight loss than dieting. We proposed an alternate strategy to target this receptor, while reducing likelihood of side effects and/or toxicity. Positive allosteric modulators (PAMs) with minimal intrinsic agonist activity would enhance CCK action, while maintaining spatial and temporal characteristics of physiologic signaling. This could correct abnormal stimulus–activity coupling observed in a high-cholesterol environment observed in obesity. We utilized high-throughput screening to identify a molecule with this pharmacological profile and studied its basis of action. Compound 1 was a weak partial agonist, with PAM activity to enhance CCK action at CCK1R, but not CCK2R, maintained in both normal and high cholesterol. Compound 1 (10 µM) did not exhibit agonist activity or stimulate internalization of CCK1R. It enhanced CCK activity by slowing the off-rate of bound hormone, increasing its binding affinity. Computational docking of Compound 1 to CCK1R yielded plausible poses. A radioiodinatable photolabile analogue retained Compound 1 pharmacology and covalently labeled CCK1R Thr211, consistent with one proposed pose. Our study identifies a novel, selective, CCK1R PAM that binds to the receptor to enhance action of CCK-8 and CCK-58 in both normal and disease-mimicking high-cholesterol environments. This facilitates the development of compounds that target the physiologic spatial and temporal engagement of CCK1R by CCK that underpins its critical role in metabolic regulation.
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影响因子：
2.9
作者：
Hadac, EM;Ghanekar, DV;Miller, LJ
通讯作者：
Miller, LJ
影响因子：
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Miller LJ
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通讯作者：
Miller, LJ
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通讯作者：
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