Small molecule chemical probes are valuable tools for interrogating protein biological functions and relevance as a therapeutic target. Rigorous validation of chemical probe parameters such as cellular potency and selectivity is critical to unequivocally linking biological and phenotypic data resulting from treatment with a chemical probe to the function of a specific target protein. A variety of modern technologies are available to evaluate cellular potency and selectivity, target engagement, and functional response biomarkers of chemical probe compounds. Here, we review these technologies and the rationales behind using them for the characterization and validation of chemical probes. In addition, large-scale phenotypic characterization of chemical probes through chemical genetic screening is increasingly leading to a wealth of information on the cellular pharmacology and disease involvement of potential therapeutic targets. Extensive compound validation approaches and integration of phenotypic information will lay foundations for further use of chemical probes in biological discovery.

中文翻译：

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验证用于生物发现的小分子化学探针

小分子化学探针是研究蛋白质生物学功能和作为治疗靶点的相关性的宝贵工具。严格验证化学探针参数（例如细胞效力和选择性）对于明确地将化学探针处理产生的生物和表型数据与特定靶蛋白的功能联系起来至关重要。有多种现代技术可用于评估化学探针化合物的细胞效力和选择性、靶标参与和功能反应生物标志物。在这里，我们回顾这些技术以及使用它们来表征和验证化学探针的基本原理。此外，通过化学遗传筛选对化学探针进行大规模表型表征，越来越多地获得有关潜在治疗靶点的细胞药理学和疾病参与的大量信息。广泛的化合物验证方法和表型信息的整合将为化学探针在生物发现中的进一步使用奠定基础。