In silico high-throughput screening system for AKT1 activators with therapeutic applications in sepsis acute lung injury

Abstract

PurposeAKT1 is an important target in sepsis acute lung injury (SALI). The current study was aim to construct a high-throughput screening (HTS) system based on the ChemDiv database (https://www.chemdiv.com/complete-list/) and use the system to screen for AKT1 activation agents, which may provide clues for the research and development of new drugs to treat SALI.MethodsBased on the existing X-ray structure of AKT1 and known AKT activators, a large-scale virtual HTS was performed on the ChemDiv database of small molecules by the cascade docking method and demonstrated both accuracy and screening efficiency. Molecular docking and molecular dynamics simulations were used to assess the stability and binding characteristics of the identified small-molecule compounds. The protective effect of the new highly selective compound on SALI were verified both in vitro and in vivo experiments.ResultsThe small-molecule compound 7460-0250 was screened out as a specific activator of AKT1. Molecular validation experiments confirmed that compound 7460-0250 specifically promoted the phosphorylation of AKT1 and down-regulated the LPS-induced apoptosis of human umbilical vein endothelial cells (HUVECs) by activating the AKT-mTOR pathway. Up-regulated mTOR was detected to directly interact with Bax to reduce apoptosis. In vivo, compound 7460-0250 could improved survival rate and alleviated lung injury of sepsis mice induced by cecum ligation and puncture (CLP), parallel with the activation of the AKT-mTOR pathway.ConclusionSmall-molecule compound 7460-0250 was successfully screened and confirmed as a highly selective AKT1 activator, which is a critical target in the development of new therapeutics for SALI.

Reduning alleviates sepsis-induced acute lung injury by reducing apoptosis of pulmonary microvascular endothelial cells

“…However, adipose-derived mesenchymal stem cells-derived exosomes have been found to inhibit lung hemorrhage and edema, and reduce vascular hyper-permeability by activating the PI3K-AKT pathway and protecting endothelial cells ( 34 ). In a previous study, it was demonstrated that mTOR can directly bind to Bax in HUVECs, highlighting the importance of studying drugs that can interfere with the PI3K-AKT-mTOR pathway for the prevention and treatment of SALI ( 18 ). Therefore, it is very important to study and explore drugs that can interfere with the PI3K/AKT/mTOR pathway for the prevention and treatment of SALI.…”

“…The elution was carried out using 1X Laemmli buffer, and the Bax protein was analyzed by WB analysis from the elution content. The protocols for SDS-PAGE and WB have been previously described ( 18 ).…”

“…Mice were aged between 6 and 8 weeks and weighted between 20 and 22 g. They were raised in a pathogen-free environment at Tsinghua University. Cecum ligation and puncture (CLP) was used to establish the SALI mice model which was already confirmed in a previous study (18). The C57BL/6J mice were divided into four groups of 12 mice each: sham, RDN, SALI, and SALI+RDN.…”