In this study we used pregabalin, diazepam and glucose for evaluating which one are more effective agents against glutamate induced neurotoxicity. After cortex culture preparation glutamate toxicity induced by adding 10-5 mM glutamate to each wells except negative control group. After 10 min different dose of diazepam, pregabalin and glucose were added for 24 h and then 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), Total Antioxidant Status (TAC) and Total oxidant status (TOS) assays were done. Diazepam, dose dependently shows neuroprotective effects and low dose of diazepam did not increase cell viability higher than 80 %. Combination groups only in high dose increased neurons viability. Total Antioxidant capacity of Pregabalin also show correlation with diazepam groups and highest dose of both show nearest value to negative control group. In addition, lowest and highest oxidant level were gained by the negative and positive control groups respectively. Except high dose of combination group, all groups show statically difference in compare with the negative control group P<0,05. According to our data combination of diazepam, pregabalin and glucose are more effective than pure drug to reduce glutamate toxicity in cortex neuron culture.