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Background: Esophageal cancer is a sixth most cause of cancer-associated mortalities worldwide with increased global prevalence in each year. It has a poor prognosis with 5-year survival rate are less than 10%. Objective: The present study was focused to fabricate the chitosan-sodium alginate-polyethylene glycol-crocin nanocomposites (CSP-Cr-NCs) and evaluate its in vitro anticancer potential against the esophageal cancer KYSE-150 cells. Methodology: The fabricated CSP-Cr-NCs were characterized using different techniques such as UV–visible spectroscopy, photoluminescence assay, DLS analysis, XRD, SEM and EDX analyses. The antimicrobial study was conducted by well diffusion technique against the S. pneumoniae, K. pneumoniae, E. coli, S. aureus, and C. albicans. The viability of CSP-Cr-NCs treated KYSE-150 and Het-1A cells were assessed by the MTT assay. The ROS production, MMP level, and apoptotic cell death in the CSP-Cr-NCs administered cells were assessed by using different fluorescent staining techniques. The cell migration of CSP-Cr-NCs treated KYSE-150 cells were assessed by wound scratch assay. The levels of TBARS, GSH, and SOD activity in the CSP-Cr-NCs treated KYSE-150 cells were assessed by kits. Results: The outcomes from the different characterization analyses witnessed the formation of CSP-Cr-NCs with the 100–210 nm size, tetragonal and agglomerated morphological appearances. The CSP-Cr-NCs effectively repressed the microbial growth. The CSP-Cr-NCs treated KYSE-150 cells were demonstrated the decreased cell viability and IC50 was found at 2.5 µg, furthermore it did not affected the normal Het-1A cells. The formulated CSP-Cr-NCs treatment at 2.5 and 5 µg improved the ROS production, and decreased the MMP status in the KYSE-150 cells. The elevated incidences of apoptotic cells death was found in the CSP-Cr-NCs treated KYSE-150 cells. The CSP-Cr-NCs also inhibited the migration of KYSE-150 cells. The increased TBARS content and decreased GSH and SOD activity was also found in the CSP-Cr-NCs treated KYSE-150 cells. Conclusion: Our findings proved that the formulated CSP-Cr-NCs treatment effectively inhibited the esophageal cancer KYSE-150 cell growth via increasing ROS production and apoptotic cell death. Therefore, it could be a promising anticancer candidate in the future for the esophageal cancer treatment.