Unit:
Κατεύθυνση Σχεδιασμός και Ανάπτυξη νέων Φαρμακευτικών Ενώσεων - ΦαρμακολογίαLibrary of the School of Science
Author:
Karagiannis Dimitrios
Supervisors info:
Μικρός Εμμανουήλ, Καθηγητής του τομέα Φαρμακευτικής Χημείας, Τμήμα Φαρμακευτικής Αθηνών
Παπαπετρόπουλος Ανδρέας, Καθηγητής του τομέα Φαρμακευτικής Χημείας, Τμήμα Φαρμακευτικής Αθηνών
Ανδρεάδου Ιωάννα, Αναπλ. Καθηγήτρια του τομέα Φαρμακευτικής Χημείας, Τμήμα Φαρμακευτικής Αθηνών
Original Title:
Προσδιορισμός νέων αναστολέων της μιτοχονδριακής ATP συνθάσης. In silico επιλογή και in vitro φαρμακολογική αξιολόγηση.
Translated title:
Identification of novel ATP synthase inhibitors. In silico screening and in vitro pharmacological evaluation.
Summary:
ATP synthase (also known as F-ATPase or F1Fo-ATPase) is a significant multiprotein complex, proven to be a small rotor machine, either producing or hydrolyzing adenosine triphosphate. During myocardial ischemia ATP synthase's rotor switch to hydrolyzing activity, wasting ATP and generating dysfunctional mitochondria. (1) Selective inhibition of ATP hydrolase, could be of major significance for cardioprotection and the discovery of new inhibiting agents is of particular interest for the development of new therapeutic schemes.
In this work, we present the identification of inhibitors of the hydrolytic activity of ATP synthase, based on virtual and experimental screening methods.
In a first step, we constructed for the first time a holistic model of ATP synthase of mus musculus based on homology modeling using all known subunits from Protein Data Bank (PDB). Based on this model, docking-scoring calculations have been performed targeting the three major inhibition sites already known from literature: a) the ATP-ADP catalytic site b) the oligomycin binding site and c) inside the surface of a groove made from loops in the 3α and 3β subunits where molecules like quercetin or resveratrol are known to bind. The workflow has been implemented on our in-house library of 2000 natural products and synthetic compounds (Pharmalab). Moreover, ligand-based virtual screening, has been carried out based on similarity calculations of the chemical library components against known binders like BMS-199264. (1)
Different steps of filtering through the in silico screening provided 48 molecules that were further tested in vitro on isolated murine heart mitochondrial fractions, evaluating ATP synthase hydrolytic activity. C57-BL-6 mice’ hearts were cut to isolate mitochondria. In order ATP synthase to function hydrolyzing ATP, a freeze-thaw cycle of the sample was followed, while the production of PO43- has been finally evaluated. The results showed that 35 out of 48 (73%) tested compounds exhibited various inhibitory activities at 0.2 mM, validating our computer-based drug design method. Among 35 hits, 7 molecules appeared to have common scaffold displaying enhanced activity (48 – 82.5% inhibition). The IC50 of these compounds has been further determined, providing confirmatory evidence of the inhibitory activity of the scaffold which can lead us to a novel agent against myocardial ischemia.
Main subject category:
Science
Other subject categories:
Pharmacology
Keywords:
mitochondrial atp synthase, atp synthase mouse, atp synthase mus musculus, atp hydrolysis inhibitors, atp hydrolysis, myocardial ischaemia, cardioprotective, mitochondrial isolation
Number of references:
127
