In the current study, the cyclopeptide alkaloid discarine D-derived tri-peptides fragments were synthesized and then investigated for their inhibitory potential against krait (Bungarus sindanus) venom acetylcholinesterase (AChE) enzyme. The tri-peptides L-Leu- threo-D-Pheser-L-Phe and L-Leu- threo-L-Pheser-L-Phe were chemically synthesized by a conventional method using the benzyloxycarbonyl group for the alpha-amino (alpha-amino) safety and the methyl esters an amino acids derivative used for the safety of carboxyl group. The present paper described that the general synthetic strategy of tri-peptide allows the tri-peptide sequence to be acquired with the N-terminal extreme protected. Kinetic studies using the Lineweaver Burk plot indicated that tri-peptides fragments cause an un-competitive type of inhibition i.e. both K-m and V-max values decreased with the increase of tri-peptides fragment concentration (13.5-22.5 mu M). The estimated K-i and IC50 for krait venom AChE were found to be 17.5 mu M and 19.5 mu M, respectively. Thus the present paper, clarified that the freshly produced tri-peptides fragment can be deliberated as a beneficial mediator for the inhibition of krait venom AChE.