Yanamandra Mahesh, Mitra Sayan, Giri Archana
Scientist, GVK Biosciences Private Ltd, Biology , Campus MLR 1, Survey Nos. 125 (part) and 126, IDA Mallapur, Hyderabad, Telangana, 500076 , India +91 9989086492 ;
Expert Opin Drug Discov. 2015 Feb;10(2):171-86. doi: 10.1517/17460441.2015.997205. Epub 2014 Dec 30.
Phosphoinositide 3-kinases (PI3Ks) constitute one of the most important signaling pathways, playing a vital role in cellular differentiation and proliferation with a key function in cellular receptor triggered signal transduction downstream of tyrosine kinase receptors and/or G-protein coupled receptors. PI3K promotes cell survival proliferation, protein synthesis and glucose metabolism by generating secondary messengers phospholipid phosphatidyl 3,4,5-triphosphate and signaling via AKT/mTOR regulation. Deregulation of PI3K pathways have been observed in cancer, diabetes, neurological and inflammatory diseases and is an attractive target for pharmaceutical industries.
In this review, the authors explain different PI3K assay methodologies. Furthermore, the authors summarize the techno-scientific principles and their utility in profiling novel chemical entities against PI3Ks. Specifically, the authors compare different PI3K assay formats explaining their mode of detection as well as their advantages and limitations for drug discovery efforts.
Developing lipid (PI3K) kinase assays involves significant effort and a rational understanding is needed due to the intrinsic lipidic nature of phospholipid phosphatidyl 4,5-biphosphate, which is used as an in vitro substrate for assays with PI3K isoforms. The assay of choice should be versatile, homogenous and definitely adaptable for high-throughput screening campaigns. Additionally, these assays are expected to dissect the mechanism of action of novel compounds (inhibitor characterization) against PI3K. Existing methods provide the versatility to medicinal chemists such that they can choose one or more assay platform to progress their compounds while profiling and/or inhibitor characterization.
磷酸肌醇3激酶（PI3Ks）构成了最重要的信号通路之一，在细胞分化和增殖中发挥着至关重要的作用，在酪氨酸激酶受体和/或G蛋白偶联受体下游的细胞受体触发信号转导中起关键作用。PI3K通过生成二级信使磷脂酰3,4,5-三磷酸并经由AKT/mTOR调节发出信号，从而促进细胞存活、增殖、蛋白质合成和葡萄糖代谢。在癌症、糖尿病、神经和炎症性疾病中已观察到PI3K通路失调，它是制药行业有吸引力的靶点。
在本综述中，作者解释了不同的PI3K检测方法。此外，作者总结了技术科学原理及其在分析针对PI3Ks的新型化学实体方面的用途。具体而言，作者比较了不同的PI3K检测形式，解释了它们的检测模式以及在药物发现工作中的优缺点。
开发脂质（PI3K）激酶检测方法需要付出巨大努力，由于用作PI3K亚型检测体外底物的磷脂酰4,5-二磷酸具有内在的脂质性质，因此需要合理的理解。所选择的检测方法应具有通用性、均一性，并且肯定适用于高通量筛选活动。此外，预计这些检测方法能够剖析新型化合物（抑制剂特性）针对PI3K的作用机制。现有方法为药物化学家提供了通用性，使他们在分析和/或抑制剂特性研究时可以选择一个或多个检测平台来推进他们的化合物研究。
