Structural biology in aid of the design of next generation PKD2 inhibitors

Multiple clinically inhibitors targeting protein kinase D2 (PKD2) are available, but all of these compounds are currently ATP-competitive and lack target sensitivity. As PKD1 has an opposing effect to the PKD2, this selectivity is essential. Recently, the host laboratory identified a series of compounds named AllStars (Allosteric Stabilizers) targeting PKD2 in an allosteric and PKD2 specific way. Therefore, the main goal of my project is to analyse and improve the potency of these AllStars in vitro and in cellullo. First, I'm optimizing a radiometric kinase assay, differential scanning fluorimetry (DSF) assay and biologic assays for analysis of the inhibitory activity of the AllStars. Secondly, I will perform in vitro and in cellullo target identification and validation via PKD isoform domain swapping and Crispr-cas9 targeted mutagenesis. Next, the target domain(s) of PKD2 will be cocrystallized with the most potent AllStars. The interactions of the AllStars with the target domain will be used for receptor based drug design to improve the AllStars and design different chemotypes of AllStars like compounds. These newly, improved, compounds will be analysed again in the kinase assay, DSF assay and biology assays as well as being co-crystallized with the PKD2 target domain. As such this project will yield several isoform specific PKD2 inhibitors representing an unique IP for further clinical development.