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High fat-high fructose diet (HFFD) lead to overnutrition which disturb glucose homeostasis. Insulin plays a pivotal role in glucose metabolism, and its role was affected during HFFD. Soybean (Glycine max L.) contains isoflavones, particularly daidzein and genistein which can be transformed using microbial and physical stimuli to synthesis glyceollin. Glyceollin is a phytoalexin which has a beneficial effect on health. The aim of this study was to evaluate the effect of elicited soybean extract (ESE) on insulin concentration in high fat-fructose diet (HFFD) mice. Twenty-eight female Balb/C mice were divided into seven groups: normal diet (ND) only, ND + ESE 104 mg/kg BW, HFFD only, HFFD + Simvastatin 2.8 mg/kg, HFFD + ESE 78 mg/kg BW, HFFD + ESE 104 mg/kg BW, and HFFD + ESE 130 mg/kg BW. The high fat-fructose diet was given over a period of 20 weeks, and ESE was administered orally per day after 20 weeks for four weeks. At week 24, the animals were sacrificed and the serum was collected. Insulin concentration was determined by indirect ELISA. The HFFD treatment significantly increased insulin concentration (p < 0.05) compared to a normal diet. The ESE treatment in HFFD mice could normalize insulin concentration compared to HFFD mice. Our results suggest that ESE has the potential to be used as a supplement to improves metabolic disorder through restore insulin concentration.