Spiranic compounds as novel AMPK paradoxical inhibitors: can we derive new mechanistic

[EN] In this study, we synthesized and fully characterized spiranic compounds designed as conformationally restricted mimetics of A-769662 by means of crystallographic, spectroscopic, and computational methods. In vitro enzymatic assays and surface plasmon resonance experiments revealed a mixed-type...

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Bibliographic Details
Authors: Sanz-Gomez, Marta, Barmpidi, Katerina, Quesada-Sanchez, Sergio, Cumella, Jose, Perez, Concepcion, Sanz, Pascual, Doyaguez, Elisa G., Infantes, Lourdes, Luque, Francisco Javier, Castro, Ana, Lagartera, Laura, Fernandez-Alfonso, Maria S., Estarellas, Carolina, García-Gimeno, María Adelaida|||0000-0002-6483-3912
Format: article
Publication Date:2026
Country:España
Institution:Universitat Politècnica de València (UPV)
Repository:RiuNet. Repositorio Institucional de la Universitat Politécnica de Valéncia
Language:English
OAI Identifier:oai:dnet:riunet______::6fca767c1e08595504d76e8e60304354
Online Access:https://riunet.upv.es/handle/10251/235054
Access Level:Open access
Keyword:AMPK
Mixed-type inhibition
Vasodilation
Endothelial function
Protein dynamics
Closed A -loop
Paradoxical inhibitor
Description
Summary:[EN] In this study, we synthesized and fully characterized spiranic compounds designed as conformationally restricted mimetics of A-769662 by means of crystallographic, spectroscopic, and computational methods. In vitro enzymatic assays and surface plasmon resonance experiments revealed a mixed-type inhibition mechanism. We developed a stable and equilibrated alpha 1(31 AMPK model featuring the A-loop in a closed conformation, which was used to investigate alternative binding pockets beyond the canonical AMPK sites, namely type II-like pocket. The cell-based and tissue experiments further demonstrated that these compounds act as paradoxical inhibitors, since it enhances downstream signalling by increasing phosphorylation of ACC and eNOS, ultimately activating the AMPK-eNOS-NO pathway and inducing vasodilation in rat aorta. Altogether, our findings offer new insights into AMPK modulation and highlight the potential of spiranic scaffolds as tools for probing alternative regulatory mechanisms.