Grånäs Charlotta, Lundholt Betina Kerstin, Heydorn Arne, Linde Viggo, Pedersen Hans-Christian, Krog-Jensen Christian, Rosenkilde Mette M, Pagliaro Len
BioImage A/S, Copenhagen, Denmark.
Comb Chem High Throughput Screen. 2005 Jun;8(4):301-9. doi: 10.2174/1386207054020741.
G protein-coupled receptors (GPCRs) have been one of the most productive classes of drug targets for several decades, and new technologies for GPCR-based discovery promise to keep this field active for years to come. While molecular screens for GPCR receptor agonist- and antagonist-based drugs will continue to be valuable discovery tools, the most exciting developments in the field involve cell-based assays for GPCR function. Some cell-based discovery strategies, such as the use of beta-arrestin as a surrogate marker for GPCR function, have already been reduced to practice, and have been used as valuable discovery tools for several years. The application of high content cell-based screening to GPCR discovery has opened up additional possibilities, such as direct tracking of GPCRs, G proteins and other signaling pathway components using intracellular translocation assays. These assays provide the capability to probe GPCR function at the cellular level with better resolution than has previously been possible, and offer practical strategies for more definitive selectivity evaluation and counter-screening in the early stages of drug discovery. The potential of cell-based translocation assays for GPCR discovery is described, and proof-of-concept data from a pilot screen with a CXCR4 assay are presented. This chemokine receptor is a highly relevant drug target which plays an important role in the pathogenesis of inflammatory disease and also has been shown to be a co-receptor for entry of HIV into cells as well as to play a role in metastasis of certain cancer cells.
几十年来，G蛋白偶联受体（GPCRs）一直是最有成效的一类药物靶点，基于GPCR的发现新技术有望在未来数年保持该领域的活跃。虽然基于GPCR受体激动剂和拮抗剂的药物的分子筛选仍将是有价值的发现工具，但该领域最令人兴奋的进展涉及基于细胞的GPCR功能检测。一些基于细胞的发现策略，如使用β-抑制蛋白作为GPCR功能的替代标志物，已经付诸实践，并作为有价值的发现工具使用了数年。基于细胞的高内涵筛选在GPCR发现中的应用开辟了更多可能性，例如使用细胞内转位检测直接追踪GPCR、G蛋白和其他信号通路成分。这些检测能够在细胞水平上以比以往更高的分辨率探测GPCR功能，并为药物发现早期阶段更明确的选择性评估和反筛选提供实用策略。本文描述了基于细胞的转位检测在GPCR发现中的潜力，并展示了用CXCR4检测进行初步筛选的概念验证数据。这种趋化因子受体是一个高度相关的药物靶点，在炎症性疾病的发病机制中起重要作用，并且已被证明是HIV进入细胞的共受体，还在某些癌细胞的转移中发挥作用。
