The present study demonstrates the fabrication of a nanocarrier system using diselenide conjugated gelatin (G-Se-Se-G) for the redox responsive delivery of a wide spectrum anticancer drug, doxorubicin (Dox). The method involves nanoprecipitation of the aqueous solution of G-Se-Se-G employing ethanol and F68 as the desolvating and stabilizing agents respectively. The drug, Dox is loaded within the nanocarrier by entrapment method during the nanoparticle synthesis. The physico-chemical characterizations confirm the formation of nanoparticles of regular spherical shape with hydrodynamic size, surface charge and loading efficiency of 181 ± 12.6 nm, -13.10 ± 0.83 mV and ∼9 % respectively. Notably, Dox loaded G-Se-Se-G carrier shows significantly higher drug release in response to acidic (pH = ∼5.5) and reducing (10 mM glutathione) sink conditions. Dox release kinetic data fitted to a universal Korsmeyer–Peppas model suggests that irrespective of the nature of stimuli, the release of Dox from G-Se-Se-G carrier follows non-Fickian mechanism which is attributed to the diffusion and erosion. The cytotoxicity studies employing human lung adenocarcinoma (A549) and human normal lung fibroblast (WI38) cells confirm the potency as well as selectively of Dox loaded G-Se-Se-G carrier towards cancer cells. The intracellular drug release studies through fluorescence imaging indeed corroborate the redox (reducing environment) mediated preferential release of Dox from G-Se-Se-G carrier in A549 cells as compared to WI38 cells. Finally, in vivo studies using A549 derived xenograft tumor model establishes that Dox loaded G-Se-Se-G nanocarrier is better tolerable than free Dox with almost similar efficacy.