Gliomas account for the majority of human brain tumors and the incidence of gliomas is expected to risein upcoming years and therefore extensive efforts have been devoted to the discovery of potent antiglioma agents. Dueto the importance of benzoxazoles for anticancer drug discovery, herein new benzoxazole-based hydrazone derivatives(3a-g) were designed and synthesized. The cytotoxic effects of the compounds on C6 rat glioma and NIH/3T3 mouseembryonic fibroblast cell lines were investigated using MTT assay. The apoptotic effects of the most selective anticanceragent were analyzed based on Annexin V-PI binding capacities in flow cytometry. The compounds were alsoinvestigated for their acetylcholinesterase (AChE) inhibitory effects using a modification of Ellman’sspectrophotometric method. In order to evaluate the compliance of the compounds to Lipinski’s rule of five, theirphysicochemical parameters were determined using Molinspiration software. N'-([1,1'-Biphenyl]-4-ylmethylene)-2-[(5-fluorobenzoxazol-2-yl)thio]acetohydrazide (3g) was found to be more effective on C6 cell line (IC50= 4.30±0.28 µg/mL)than mitoxantrone (IC50= 4.56±1.24 µg/mL). The high SI value of compound 3g indicated that its antiglioma activitywas selective. This compound caused late apoptosis in a dose dependent manner. Compound 3g was also found to bethe most effective AChE inhibitor in this series. According to in silico studies, compound 3g only violated one parameterof Lipinski’s rule of five. On the basis of Lipinski’s rule, the compound was expected to have reasonable oralbioavailability. According to in vitro and in silico studies, compound 3g stands out as a promising antiglioma agent forfurther studies.