β-Sitosterol, a phytocompound from Parthenium hysterophorus, reveals anti-diabetic properties through α-Amylase inhibition: an in-silico and in-vitro analysis

Abstract

The study aims to identify and validate a potential α-Amylase inhibitor from the leaf extract of the Parthenium hysterophorus. Molecular docking and dynamics analyses were performed to test the anti-diabetic efficacy of the compound by focusing on α-Amylase inhibition. The molecular docking study using AutoDock Vina (PyRx) and SeeSAR tools identified β-Sitosterol as an effective α-Amylase inhibitory compound. Among the analysed fifteen phytochemicals, β-Sitosterol demonstrated the most appreciable binding energy (-9.0 Kcal/mol) and is comparatively higher than the binding energy of the standard α-Amylase inhibitor, the Acarbose (-7.6 Kcal/mol). The significance of the interaction between β-Sitosterol and α-Amylase was further investigated using Molecular Dynamics Simulation (MDS) for 100 ns via GROMACS. The data reveals that the compound could exhibit the highest stability with α-Amylase regarding RMSD, RMSF, SASA and Potential Energy analysis. The key residue of α-Amylase (Asp −197) shows a significantly low fluctuation of 0.7 Å while interacting with β-Sitosterol. The data obtained from MDS results strongly suggested the potential inhibitory impact of β-Sitosterol on α-Amylase. In addition, the proposed phytochemical was purified from the leaf extracts of P.hysterophorus using the silica gel column chromatography and identified by GC-MS analysis. The purified β-Sitosterol demonstrated a significant 42.30% in-vitro α-Amylase enzyme inhibition property under 400 µg/ml concentration and thus supported the in-silico predictions. Further in-vivo investigations are necessary to analyse the efficiency of β-Sitosterol on α-Amylase inhibition to help the anti-diabetic potential of the phytocompound.

Communicated by Ramaswamy H. Sarma

Keywords:

• Parthenium hysterophorus
• β-Sitosterol
• anti-diabetic
• α-Amylase
• enzyme inhibition
• molecular docking
• molecular dynamic simulation

Acknowledgement

The authors would like to thank Fr. Dr. Victor Lobo SJ (Vice-Chancellor), Fr. Denzil E. Lobo SJ (Director of Information Technology), and Fr. Dr. S Xavier SJ (Director of Research) of St. Joseph’s College (Autonomous), Bangalore for providing Workstation and SeeSAR tool for carrying out this research work.

Author contribution

Lokesh Ravi—Designed the work and analysed the data

Shabari Girish—Performed molecular docking

Sharun Roy D’Souza- Performed molecular dynamics

Anirudh Sreenivas BK- Critically evaluated the manuscript

Shree Kumari GR- Critically evaluated the manuscript

Reji Manjunathan -Designed the work, analysed the data and wrote the manuscript.

Consent for publication

All authors have provided consent to publish this manuscript in your esteemed journal.

Disclosure statement

No potential conflict of interest was reported by the authors.

Additional information

Funding

The seed Money Project Grant provided by St. Joseph’s College (Autonomous), Bengaluru, provided basic facilities to execute this research work.