Neuroblastoma (NB) may be the common pediatric tumor from the supportive central nervous system characterised by poor prognosis. Because of the difficulties for example high tumor heterogeneity, multidrug resistance, minimal residual disease, etc., there’s an instantaneous requirement for exploring new therapeutic strategies and efficient treating NB. Herein, in the present study, we explored the untouched response of NB cells towards the second-generation histone deacetylase inhibitor (HDACi) JNJ-26481585(JNJ) and also the lysosomotropic agent, Chloroquine (CQ) alone and upon JNJ/CQ treatment like a plausible therapeutic. We see that while JNJ alone caused autophagy in NB cells, JNJ/CQ treatment decreased the viability and proliferation of NB cells in vitro by switching from autophagy to apoptosis. Further we discovered that autophagy inhibition by CQ pre-treatment brought towards the generation of ROS and home loan business the mitochondrial membrane potential (MMP) that subsequently caused caspase-3-mediated apoptotic cell dying in NB cells. Corroborating the above mentioned observations, we discovered that the ROS scavenger N-acetylcysteine (NAC) countered caspase-3 activity and also the cells were saved from apoptosis. Finally, these observations establish that JNJ/CQ treatment led to cell dying in NB cells by triggering the development of ROS and disruption of MMP, suggesting that modulation of JNJ-caused autophagy by CQ represents an encouraging new therapeutic approach in NB.