Hepsin,a transmembrane serine protease abundant in renal endothelial cells,is a promising therapeutic target against several cancers,particularly prostate cancer.It is involved in the release and polymerization of uromodulin in the urine,which plays a role in kidney stone formation.In this work,we design new potential hepsin inhibitors for high activity,improved specificity towards hepsin,and promising ADMET properties.The ligands were developed in silico through a novel hierarchical pipeline.This pipeline explicitly accounts for off-target binding to the related serine proteases matriptase and HGFA (human hepatocyte growth factor activator).We completed the pipeline incorporating AD-MET properties of the candidate inhibitors into custom multi-objective optimization functions.The ligands designed show excellent prospects for targeting hepsin via the blood stream and the urine and thus enable key experimental studies.The computational pipeline proposed is remarkably cost-efficient and can be easily adapted for designing inhibitors against new drug targets.