The aim of the present study was to develop and characterize targeted sustained release nanoparticles of chitosan-glutamic acid conjugates containing triple therapy for Helicobacter pylori to improve its therapeutic effect and reduce its dose-related side effect. The chitosan-glutamate nanoparticles were prepared by using the ionotropic gelation method. The particle sizes of systems ranged between 550 nm and 900 nm. Percent drug entrapment and release profiles of amoxicillin, clarithromycin, and omeprazole in simulated gastric fluid (pH 1.2) were determined using high-performance liquid chromatography. The maximum % drug entrapment and % yield of formulations were about 60-90% and 73-88%, respectively. The stability of the drugs was assessed in simulated gastric fluid (pH 1.2). In vitro antibacterial efficacy of optimized formulations containing monotherapy and triple therapy on isolated culture of H. pylori was assessed. In vivo clearance study and histopathological study were also carried out on Swiss albino mice to evaluate the efficacy of triple therapy containing targeted nanosystem for the treatment of H. pylori. The proposed delivery system holds promise at optimizing the treatment of H. pylori.