Morris Lindsey C, Days Emily L, Turney Maxine, Mi Dehui, Lindsley Craig W, Weaver C David, Niswender Kevin D
Department of Medicine, Division of Diabetes, Endocrinology, and Metabolism, Vanderbilt University School of Medicine, Nashville, TN, USA.
Vanderbilt Institute for Chemical Biology, Vanderbilt University School of Medicine, Nashville, TN, USA.
J Biomol Screen. 2014 Jul;19(6):847-58. doi: 10.1177/1087057114520971. Epub 2014 Feb 13.
Injectable, degradation-resistant peptide agonists for the glucagon-like peptide 1 (GLP-1) receptor (GLP-1R), such as exenatide and liraglutide, activate the GLP-1R via a complex orthosteric-binding site and are effective therapeutics for glycemic control in type 2 diabetes. Orally bioavailable orthosteric small-molecule agonists are unlikely to be developed, whereas positive allosteric modulators (PAMs) may offer an improved therapeutic profile. We hypothesize that allosteric modulators of the GLP-1R would increase the potency and efficacy of native GLP-1 in a spatial and temporally preserved manner and/or may improve efficacy or side effects of injectable analogs. We report the design, optimization, and initial results of a duplexed high-throughput screen in which cell lines overexpressing either the GLP-1R or the glucagon receptor were coplated, loaded with a calcium-sensitive dye, and probed in a three-phase assay to identify agonists, antagonists, and potentiators of GLP-1, and potentiators of glucagon. 175,000 compounds were initially screened, and progression through secondary assays yielded 98 compounds with a variety of activities at the GLP-1R. Here, we describe five compounds possessing different patterns of modulation of the GLP-1R. These data uncover PAMs that may offer a drug-development pathway to enhancing in vivo efficacy of both endogenous GLP-1 and peptide analogs.
用于胰高血糖素样肽1（GLP-1）受体（GLP-1R）的可注射、抗降解肽激动剂，如艾塞那肽和利拉鲁肽，通过复杂的正构结合位点激活GLP-1R，是2型糖尿病血糖控制的有效治疗药物。口服生物可利用的正构小分子激动剂不太可能被开发出来，而正变构调节剂（PAMs）可能具有更好的治疗效果。我们假设，GLP-1R的变构调节剂将以空间和时间上保留的方式提高天然GLP-1的效力和功效，和/或可能改善可注射类似物的疗效或副作用。我们报告了一种双重高通量筛选的设计、优化和初步结果，在该筛选中，将过表达GLP-1R或胰高血糖素受体的细胞系共培养，加载钙敏染料，并在三相试验中进行检测，以鉴定GLP-1的激动剂、拮抗剂和增效剂，以及胰高血糖素的增效剂。最初筛选了175,000种化合物，通过二次试验得到了98种在GLP-1R上具有多种活性的化合物。在这里，我们描述了五种对GLP-1R具有不同调节模式的化合物。这些数据揭示了可能为增强内源性GLP-1和肽类似物的体内疗效提供药物开发途径的PAMs。
