Cholelithiasis poses significant health and economic burdens, necessitating novel pharmacological targets to enhance treatment efficacy.Based on genome-wide association analysis (GWAS) studies, we performed two-sample Mendelian randomization (MR) analysis based on plasma proteomics to explore potential drug targets in European (nCase=40,191 and nControl=361,641) and Asian (nCase=9,305 and nControl=168,253) populations. We confirmed the directionality and robust correlation of the drug targets with the results through reverse MR analysis, Steiger filtering, Bayesian colocalization, phenotype scanning and replication in multiple databases. Further exploration of the safety and possible mechanisms of action of phenome-wide MR analysis and protein‒protein interactions (PPIs) as individual drug targets was performed.Our proteomics-based MR analyses suggested that FUT3 (OR=0.87; 95% CI, 0.84-0.89; P=4.70×10-32), NOE1 (OR=0.58; 95% CI, 0.52-0.66; P=4.21×10-23), UGT1A6 (OR=0.68; 95% CI, 0.64-0.73; P=9.58×10-30) and FKBP52 (OR=1.75; 95% CI, 1.37-2.24; P=8.61×10-6) were potential drug targets in Europeans, whereas KLB (OR=1.11; 95% CI, 1.07-1.16; P=7.59×10-7) and FGFR4 (OR=0.94; 95% CI, 0.91-0.96; P=4.07×10-6) were valid targets in East Asians. There was no reverse causality for these drug targets. Evidence from Bayesian colocalization analyses supported that exposure and outcome shared consistent genetic variables. Phenome-wide MR analysis suggested the potential deleterious effects of NOE1 and FGFR4. PPI analysis confirmed the pathways associated with the potential targets involved in bile acid metabolism.Genetically predicted levels of the plasma proteins FUT3, NOE1, UGT1A6 and FKBP52 have potential as prospective targets in Europeans. Moreover, the plasma levels of KLB and FGFR4 may serve as potential targets for the treatment of cholelithiasis in East Asians.Copyright © 2024 The Author(s). Published by Wolters Kluwer Health, Inc.