Objective: This study investigates for the first time the antidiabetic activity of 4-hydroxy isoleucine (4-OH Ile) in combination with natural bioavailability enhancers, piperine and ginger oleoresin. Methods: Alloxan induced diabetic rat model was used for studying the antidiabetic activity of 4-OH Ile in combination with natural bioavailability enhancers to study its effect on body weight, fasting blood glucose level and oral glucose tolerance test. Results: 4-OH Ile present in Trigonella foenum-graceum seeds is an amino acid that shows a significant activity in type II diabetes. Due to its glucose dependent effect and devoid of side effects, 4-OH Ile is used for management of type II diabetes. The antidiabetic activity of 4-OH Ile was found to be significantly increased in combination with piperine and ginger oleo resin. However, piperine has demonstrated better effect as compared to ginger oleoresin. Histopathological studies of pancreas were also done to further verify the effect of natural bioenhancers. Conclusion: The results of this study places piperine and ginger oleo resin as a mode of increasing activity of 4-OH Ile and thereby reducing their cost of the treatment.