COMPUTATIONAL INVESTIGATION OF IMPORTANT PHYTOCOMPOUNDS AND IN SILICO ANTIDIABETIC ACTIVITY IDENTIFIED FROM PENTATROPIS MICROPHYLLA

Abstract

AIM: Computational Investigation of important phytocompounds and in silico antidiabetic activity identified from  Pentatropis microphylla.

INTRODUCTION: Pentatropis microphylla, a plant species of interest, has attracted attention for its potential therapeutic properties, including possible anti-diabetic effects. Computational investigations offer a valuable approach to explore the important phytocompounds present in Pentatropis microphylla and their potential anti-diabetic activity. Through techniques like molecular docking, molecular dynamics simulations, and virtual screening, computational studies can predict the interactions between these phytocompounds and key proteins involved in diabetes, providing insights into their binding affinity and mode of action.

MATERIALS AND METHODS:

Invitro Alpha-amylase inhibitory experiment: In a 100 ul reaction, 1 um of test extract and 100 ml of pH 6.9 phosphate buffer were combined with 100 ml of a 1% starch solution. Controls replaced 200ul of enzyme with buffer. After 5 minutes, 500ul of dinitrosalicylic acid reagent was added. Boiling for 5 minutes followed. Absorbance at 540 nm was measured, and % inhibition was calculated using [(Control - Test)/Control]*100, with reference to acarbose.

Cell viability assay on Pentatropis microphilla: To assess biosafety and ethanol cytotoxicity, we employed the MTT and NRU assays. HepG2 cells were exposed to various plant extract concentrations (10-1000 mg/ml) and ethanol (50-1000 mM) for 24 hours. The MTT assay determined cell viability: cells (1 x 10^4) adhered for 24 hours, followed by the addition of MTT (5 mg/ml in PBS) and a 4-hour incubation. After discarding supernatants, we added 200 µl of DMSO to each well, gently mixed, and measured the developed color's absorbance at 550 nm using a multiwell microplate reader (Thermo Scientific, Vantaa, Finland). Untreated sets served as controls for accurate assessment.

Molecular docking: Molecular docking was conducted to identify potential inhibitors of MMP8 based on binding energy. Phytochemicals from various plants, obtained from PubChem-NCBI database in SD format, were converted to PDB format using OpenBabel 2.3.1. Acarbose served as the control. The 3D structure of Alpha amylase (PDB ID: 2QV4) was acquired from the Protein Data Bank, with water molecules removed. Phytocompounds were individually docked with Alpha amylase using Hex 8.0.0 protein docking program, employing FFT-based analytics for rigid docking in 6D analysis. More negative E-total values indicated stronger ligand-receptor interactions, potentially activating receptor activity, visualized in Pymol.

RESULTS: In a computational study, crucial phytocompounds from Pentatropis microphylla exhibited promising in silico antidiabetic activity. Docking analysis with Alpha amylase (PDB ID: 2QV4) revealed strong ligand-receptor interactions, indicating their potential as inhibitors. These findings suggest that Pentatropis microphylla may contain bioactive compounds with the ability to combat diabetes, warranting further experimental validation.

DISCUSSION: Computational investigation of Pentatropis microphylla's phytocompounds for in silico anti-diabetic activity involves virtual screening, molecular docking, and predictive modeling. This approach holds promise in discovering novel anti-diabetic compounds, aiding drug development, and providing insights into their mechanisms of action, accelerating diabetes research and potential therapeutic interventions.

CONCLUSION: In silico computational analysis of phytocompounds from Pentatropis microphylla revealed promising anti-diabetic potential, with strong binding affinities towards key diabetes-related targets. These findings suggest the potential efficacy of Pentatropis microphylla as a source of natural anti-diabetic agents, warranting further experimental validation and exploration for diabetes management.