Wei Huijun, Barrow James C
Department of Pharmacology and Molecular Sciences, Johns Hopkins School of Medicine, Baltimore, MD, USA.
Lieber Institute for Brain Development, Baltimore, MD, USA.
Methods Mol Biol. 2025;2972:267-274. doi: 10.1007/978-1-0716-4799-8_19.
Inositol pyrophosphates, such as 5-diphosphoinositol pentakisphosphate (IP7), play crucial roles in various biological processes. Pharmacologic inhibition of IP6K has potential therapeutic benefits for treating type II diabetes, venous thrombosis, chronic kidney disease, and psychiatric disorders. This chapter describes the identification of IP6K inhibitors through kinase-activity-based high-throughput assays and dose-response assay. Additionally, it details cell-based assays on the determination of the effect of IP6K inhibitors on IP7 production in HCT116 cells and the binding of IP6K inhibitors to IP6K inside cells through cellular thermal shift assays (CETSAs). These robust assays provide a foundation for further exploration of IP6K inhibitors in various disease states.
肌醇焦磷酸，如5-二磷酸肌醇五磷酸（IP7），在各种生物过程中发挥着关键作用。IP6K的药理学抑制对于治疗II型糖尿病、静脉血栓形成、慢性肾病和精神疾病具有潜在的治疗益处。本章描述了通过基于激酶活性的高通量测定和剂量反应测定来鉴定IP6K抑制剂。此外，还详细介绍了基于细胞的测定，即通过细胞热位移测定（CETSA）来确定IP6K抑制剂对HCT116细胞中IP7产生的影响以及IP6K抑制剂与细胞内IP6K的结合。这些可靠的测定为进一步探索IP6K抑制剂在各种疾病状态下的作用奠定了基础。
