TEAD-IN-2 (TEAD antagonist 2) is a small molecule TEAD antagonist (TEAD2 IC50=603 nM FP assays, SPR Kd=229 nM) that binds the TEAD lipid pocket and disrupts TEAD S-palmitoylation.
TEAD-IN-2 showed similar IC50 against all TEAD paralogs TEAD1, TEAD3, and TEAD4 both in IP and SRP assays.
TEAD-IN-2 disrupted Hippo signaling in a Detroit X1 562 cell reporter assay with IC50 of 31.8 nM, with no significant cytotoxicity indications in either primary human hepatocytes or human liver microtissues at 100 uM.
The inhibition of TEAD activity via TEAD-IN-2 was not due to the inability of TEAD to form a complex with YAP in cells.
TEAD-IN-2 functioned by acting as a TEAD S-palmitoylation mimic, binding and stabilizing newly translated TEAD proteins, TEAD-palmitoylation-deficient mutant K357A caused a stronger rescue of TEAD expression in HEK293T cells transfected TEAD2-K357A.
Dysregulating TEAD S-palmitoylation via TEAD-IN-2 could effectively inhibit Hippo-dependent tumor growth in vivo.