DRUG-LIKE MODULATORS TARGETING O-GLYCOSYLATION BY GalNAc TRANSFERASE-2/3

摘要 在高尔基复合体中启动位点特异性O-糖基化的大酶家族的类药物调节剂 (UDP-N-乙酰-α-D-半乳糖胺多肽N-乙酰-半乳糖胺转移酶或GalNAc-ts) 有望成为治疗骨质疏松、血脂异常、心脏病等重大疾病的全新疗法， 慢性阻塞性肺病、癌症和病毒暴发。值得注意的是，目前尚无已知的 这些起始酶的抑制物或激活剂。为了解决这一缺点，我们开发了基于细胞的 荧光传感器将用于高通量筛选以识别特定异构体的小分子 GalNAc-T2和GalNAcT3的调节剂介导O-糖基化。我们将进行同步筛查 具有这些传感器的化合物，这将大大减少偏离目标的影响，从而能够识别 直接针对任何一种酶的候选者。包括试点筛查在内的初步工作已经产生了几个 有希望的候选人辩称，进一步的筛选将是成功的。将使用以下内容验证命中率 其他试验，用纯化的酶制剂进行生化分析。在这项提议的范围之外，我们计划 任何先导化合物的结构表征和优化以及治疗价值的初步测试。 异构体特异性调节子的成功识别有望对糖生物学研究产生变革， 可能还有诊所。

SUMMARY Drug-like modulators of the large enzyme family that initiates site-specific O-glycosylation in the Golgi complex (UDP-N-acetyl-α-D-galactosamine polypeptide N-acetyl-galactosaminyltransferases or GalNAc-Ts) hold promise as entirely new therapeutics for major diseases such as osteoporosis, dyslipidemia, heart disease, chronic obstructive lung disease, cancer, and viral outbreaks. Significantly, there are currently no known inhibitors or activators of these initiating enzymes. To address this shortcoming we developed cell-based fluorescent sensors to be used for high-throughput screening to identify isoform-specific, small molecule modulators of GalNAc-T2 and GalNAcT3 mediated O-glycosylation. We will carryout simultaneous screening of compounds with these sensors, which will greatly minimize off-target effects allowing identification of candidates that directly target either enzyme. Preliminary work including a pilot screen has resulted in a few promising candidates arguing that further screening will be successful. Hits will be validated using, among other tests, biochemical assays with purified enzyme preparations. Beyond the scope of this proposal, we plan structural characterization and optimization of any lead compounds as well as initial tests of therapeutic value. Successful identification of isoform-specific modulators promises to be transformative to glycobiology research, and potentially, the clinic.