
==== Front
bioRxiv
BIORXIV
bioRxiv
2692-8205
Cold Spring Harbor Laboratory

10.1101/2024.09.05.611490
preprint
1
Article
AI-Based Discovery and CryoEM Structural Elucidation of a K ATP Channel Pharmacochaperone
ElSheikh Assmaa http://orcid.org/0000-0001-7360-9309

Driggers Camden M. http://orcid.org/0000-0002-2105-7175

Truong Ha H.
Yang Zhongying http://orcid.org/0000-0003-4686-0869

Allen John
Henriksen Niel
Walczewska-Szewc Katarzyna http://orcid.org/0000-0001-6422-6138

Shyng Show-Ling http://orcid.org/0000-0002-8230-8820

05 9 2024
2024.09.05.611490https://creativecommons.org/licenses/by-nc/4.0/ This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License, which allows reusers to distribute, remix, adapt, and build upon the material in any medium or format for noncommercial purposes only, and only so long as attribution is given to the creator.
http://biorxiv.org/lookup/doi/10.1101/2024.09.05.611490
nihpp-2024.09.05.611490.pdf
Abstract

Pancreatic K ATP channel trafficking defects underlie congenital hyperinsulinism (CHI) cases unresponsive to the K ATP channel opener diazoxide, the mainstay medical therapy for CHI. Current clinically used K ATP channel inhibitors have been shown to act as pharmacochaperones and restore surface expression of trafficking mutants; however, their therapeutic utility for K ATP trafficking impaired CHI is hindered by high-affinity binding, which limits functional recovery of rescued channels. Recent structural studies of K ATP channels employing cryo-electron microscopy (cryoEM) have revealed a promiscuous pocket where several known K ATP pharmacochaperones bind. The structural knowledge provides a framework for discovering K ATP channel pharmacochaperones with desired reversible inhibitory effects to permit functional recovery of rescued channels. Using an AI-based virtual screening technology AtomNet® followed by functional validation, we identified a novel compound, termed Aekatperone, which exhibits chaperoning effects on K ATP channel trafficking mutations. Aekatperone reversibly inhibits K ATP channel activity with a half-maximal inhibitory concentration (IC 50 ) ~ 9 μM. Mutant channels rescued to the cell surface by Aekatperone showed functional recovery upon washout of the compound. CryoEM structure of K ATP bound to Aekatperone revealed distinct binding features compared to known high affinity inhibitor pharmacochaperones. Our findings unveil a K ATP pharmacochaperone enabling functional recovery of rescued channels as a promising therapeutic for CHI caused by K ATP trafficking defects.
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