Introduction: This work aims to develop an efficient green synthetic procedure and cytotoxicity evaluationofbenzoxazolederivatives,anessentialbuildingblockinmedicinalchemistryresearch owing to its broad range of pharmacological properties, especially anticancer activity. Methods: The microwave-assisted condensation reaction between 2-amino-4-methylphenol and aromatic aldehydesusingiodineastheoxidantundersolvent-freeconditionswasexplored. ThecytotoxicityofthepreparedbenzoxazolesagainstthethreehumancancercelllinesincludingLu-1,Hep-G2 andMCF-7wereexaminedusingaMTTassay. Results: Five2,5-disubstitutedbenzoxazolederivativeshavebeensuccessfullysynthesizedingoodyields(67-90%)usingiodineastheoxidantunder solvent-freemicrowaveirradiationconditions. Theinitialcytotoxicityevaluationofthefoursynthesizedcompoundsrevealedthatcompounds3band3eexhibitedgoodactivityagainstMCF-7(IC50 =12 µ g/mL)andHep-G2(IC50 =17.9 µ g/mL),respectively. Remarkably,compound3cshowedthe best cytotoxicity against MCF-7 at IC50 = 4 µ g/mL. Conclusion: Microwave irradiation effectively assistedthe condensation reaction between2-amino-4-methylphenol and aromatic aldehydesin thepresenceofiodineundersolvent-freeconditionstofacilitatethedesiredbenzoxazolesingood total yields. The prepared benzoxazoles proved to be potent candidates for further study in the developmentofanticanceragents.