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Preliminary characterization and in silico studies on the alpha- amylase inhibitors from Momordica charantia AMP06 methanolic leaf extract

Philippine Journal of Biochemistry and Molecular Biology
2020; 1(1&2): 61-70
Authors
Arra B. Asejo, Patrick G. Moreno, Junie Billones, Ruel Nacario, Francisco M. Heralde III
Abstract
Regulating the activity of alpha-amylase, can help maintain blood glucose to normal levels especially among diabetics. Momordica charantia, known as ampalaya in the Philippines, was previously shown to have alpha-amylase inhibitory activity in our laboratory where among the nine locally available varieties, AMP06 gave the highest activity. This study aimed to characterize the alpha- amylase inhibitory principle in the leaf extract of AMP06 and perform in silico docking studies on putatively identified compounds. Crude methanolic leaf extract (CMLE) of M. charantia (AMP06) was prepared, subjected to sequential solvent extraction, concentrated by rotary evaporation, and tested for alpha-amylase inhibition. CMLE was shown to have inhibitory activity against alpha-amylase with values ranging from 43.53% - 65.70% at 500 ug/ mL to 25 ug/mL respectively. The sequential solvent partitioning of CMLE yielded fractions (i.e., hexane (HF), chloroform (CF), ethyl acetate (EAF) and butanol (BF) with decreased inhibitory activities that ranged from 20.12% to 38.02% compared with the CMLE. Fraction mixture of 1:1 ratio was prepared and evaluated and showed a recovery of inhibitory activity of 29.88% to 54.98%, indicating a possible synergistic activity. The individual fractions from the best fraction mixture were analyzed using LCMS/MS to determine the putative identity of the compounds which were then subjected to molecular docking to predict possible synergistic activity. Utilizing acarbose as a point of reference, as well as the top five hits from the docking experiment, in silico docking done with the compounds in the active site alone and in three allosteric sites, showed stronger binding affinity values in the active site of the enzyme. In conclusion, this study characterized the alpha-amylase inhibitory principle of M. charantia and its possible mechanism, that there may be compounds present in the extract that works synergistically in inhibiting alpha-amylase.
KEYWORDS: Alpha-amylase, Momordica charantia, Ampalaya, synergistic activity
To cite: Asejo AB, Moreno PG, Billones J, Nacario R and Heralde FIIIM.2020. Preliminary characterization and in silico studies on the alpha- amylase inhibitors from Momordica charantia AMP-06 methanolic leaf extract. PJBMB. Vol. I, No. 1, 2020, pp. 61-70. doi: 10.5555/ pjbmb.ph.2020.01.01.61
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