In Silico and In Vitro Approach to Assess Direct Allosteric AMPK Activators from Nature^{[ # ]}

Abstract

The 5′-adenosine monophosphate-activated protein kinase (AMPK) is an important metabolic regulator. Its allosteric drug and metabolite binding (ADaM) site was identified as an attractive target for direct AMPK activation and holds promise as a novel mechanism for the treatment of metabolic diseases. With the exception of lusianthridin and salicylic acid, no natural product (NP) is reported so far to directly target the ADaM site. For the streamlined assessment of direct AMPK activators from the pool of NPs, an integrated workflow using in silico and in vitro methods was applied. Virtual screening combining a 3D shape-based approach and docking identified 21 NPs and NP-like molecules that could potentially activate AMPK. The compounds were purchased and tested in an in vitro AMPK α ₁ β ₁ γ ₁ kinase assay. Two NP-like virtual hits were identified, which, at 30 µM concentration, caused a 1.65-fold (± 0.24) and a 1.58-fold (± 0.17) activation of AMPK, respectively. Intriguingly, using two different evaluation methods, we could not confirm the bioactivity of the supposed AMPK activator lusianthridin, which rebuts earlier reports.

Key words

AMP-activated protein kinase - virtual screening - 3D similarity screening - lusianthridin

^# Dedicated to Professor Dr. A. Douglas Kinghorn on the occasion of his 75th birthday.

Publication History

Received: 17 December 2021

Accepted after revision: 14 March 2022

Article published online:
01 August 2022

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