Sustained vascular smooth muscle hypercontractility promotes hypertension and 
cardiovascular disease. The etiology of hypercontractility is not completely 
understood. New therapeutic targets remain vitally important for drug discovery. 
Here we report that Pim kinases, in combination with DAPK3, regulate 
contractility and control hypertension. Using a co-crystal structure of lead 
molecule (HS38) in complex with DAPK3, a dual Pim/DAPK3 inhibitor (HS56) and 
selective DAPK3 inhibitors (HS94 and HS148) were developed to provide 
mechanistic insight into the polypharmacology of hypertension. In vitro and 
ex vivo studies indicated that Pim kinases directly phosphorylate smooth muscle 
targets and that Pim/DAPK3 inhibition, unlike selective DAPK3 inhibition, 
significantly reduces contractility. In vivo, HS56 decreased blood pressure in 
spontaneously hypertensive mice in a dose-dependent manner without affecting 
heart rate. These findings suggest including Pim kinase inhibition within a 
multi-target engagement strategy for hypertension management. HS56 represents a 
significant step in the development of molecularly targeted antihypertensive 
medications.