Design And Synthesis Of Small Molecule Inhibitors Targeting Transcription Factor TEADs

Covalent inhibitors work by rapidly inactivating electrophilic groups by forming covalent bonds with nucleophilic groups in the target protein.Compared with traditional non-covalent inhibitor drugs,it has the advantages of lower dosage,less side effects,longer duration of drug action,better tolerance and the ability to solve some hard-to-target pathways.In recent years,covalent drugs have achieved great success in the targeted EGFR,BTK and other tumor therapy fields with very good efficacy.Covalent inhibitors also solve the proprietary problem of KRAS pathway and show great potential in the emerging small molecule induced protein degradation technology.Therefore,we used this method to design and synthesize a series of covalent compounds targeting TEADs proteins,and explored their biological properties in vitro and in vivo.Hippo pathway is highly conserved in evolution.During development,it controls cell-cell contact,cell proliferation and organ size by adjusting the changes of cell microenvironment,which plays a key role in cell growth and survival.Relevant literatures have reported that this pathway is one of the potential anticancer targets.Transcription enhancer associated domain family members TEADs protein is a key downstream effector molecule of this pathway,and its self-palmitoylation pocket in the center is an ideal drug target site.Therefore,the research and design of inhibitors targeting TEADs can bring a new approach to the treatment of tumors.In the second chapter,we used the method of computer-aided drug design to conduct virtual docking between the target protein and the compound library molecule,from which we screened the Hit-12 compound,which was twice as potent as the reported fluorophenolic acid,and verified its targeting.After further analysis of its mode of action in the TEAD pocket,the Micheal receptor was introduced into the amino group of the Hit-12 compound to bind to the cysteine sulfhydryl group,which greatly improved the potency.In the further structure-activity relationship and chemical modification of inhibitors,we found the novelest potent compound LY-P24(IC₅₀=190nM±19 nM),and the thiol reaction specificity and kinase selectivity were ideal.It's also the first reported inhibitor that has a drug effect in animals.In the third chapter,we adopted the method of Scaffold Hopping,with the skeleton of LY-P24 compound and palmittic acid as the reference objects,and screened the new structure of LY-S01 compound from the compound library,showed ideal activity to TEAD1(IC₅₀=613 nM)and TEAD1/3 selectivity.Through structural modification to further enhance its interaction with TEAD3 tyrosine residues,we found that the first compound LY-S05 is currently the most potent and selective compound targeting TEAD3(IC₅₀=200 nM),and the selectivity against other protein subtypes is more than200 folds.In summary,the covalent inhibitors LY-P24 and LY-S05 targeted TEADs showed ideal activity and selectivity in both in vitro and in vivo experiments,providing a new choice and direction for targeted Hippo pathway in future cancer treatment.