Purinergic P2X receptors: Structural models and analysis of ligand-target interaction

Ben, Diego Dal; Buccioni, Michela; Lambertucci, Catia; Marucci, Gabriella; Thomas, Ajiroghene; Volpini, Rosaria

doi:10.1016/j.ejmech.2014.10.071

Cited by 39 publications

(26 citation statements)

References 101 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Homology modelling studies of the human P2X7 (hP2X7) were carried out using the closed state zebrafish P2X4 (zP2X4) receptor as a template, as described in Dal Ben [15]. Molecular Operating Environment (MOE by C.C.G., Montreal, CA, version 2012.10) suite was employed for this task.…”

Section: Methodsmentioning

confidence: 99%

“…This includes the discovery that an N -quinoline substituent results in higher binding affinity compounds compared with analogues with smaller or less hydrophilic aromatic rings [10, 11], and that a 1-carbon guanidine-phenyl linker is more desirable than longer and more functionalised analogues [12–14]. Instead, the cyanoguanidine core is thought to provide rigidity to the molecular structure by hydrogen bonding with the receptor-binding pocket [15]. Hence, compound EFB is ideally placed to display an optimal fit into the binding pocket while featuring a fluorine-18 label.…”

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Design, synthesis and evaluation in an LPS rodent model of neuroinflammation of a novel 18F-labelled PET tracer targeting P2X7

et al. 2017

Self Cite

View full text Add to dashboard Cite

BackgroundThe P2X7 receptor has been shown to play a fundamental role in the initiation and sustenance of the inflammatory cascade. The development of a novel fluorine-18 PET tracer superior and with a longer half-life to those currently available is a promising step towards harnessing the therapeutic and diagnostic potential offered by this target. Inspired by the known antagonist A-804598, the present study outlines the design via molecular docking, synthesis and biological evaluation of the novel P2X7 tracer [18F]EFB. The tracer was radiolabelled via a three-step procedure, in vitro binding assessed in P2X7-transfected HEK293 and in B16 cells by calcium influx assays and an initial preclinical evaluation was performed in a lipopolysaccharide (LPS)-injected rat model of neuroinflammation.ResultsThe novel tracer [18F]EFB was synthesised in 210 min in 3–5% decay-corrected radiochemical yield (DC RCY), >99% radiochemical purity (RCP) and >300 GBq/μmol and fully characterised. Functional assays showed that the compound binds with nM K i to human, rat and mouse P2X7 receptors. In vivo, [18F]EFB displayed a desirable distribution profile, and while it showed low blood–brain barrier penetration, brain uptake was quantifiable and displayed significantly higher mean longitudinal uptake in inflamed versus control rat CNS regions.Conclusions[18F]EFB demonstrates strong in vitro affinity to human and rodent P2X7 and limited yet quantifiable BBB penetration. Considering the initial promising in vivo data in an LPS rat model with elevated P2X7 expression, this work constitutes an important step in the development of a radiotracer useful for the diagnosis and monitoring of clinical disorders with associated neuroinflammatory processes.Electronic supplementary materialThe online version of this article (doi:10.1186/s13550-017-0275-2) contains supplementary material, which is available to authorized users.

show abstract

Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Design, synthesis and evaluation in an LPS rodent model of neuroinflammation of a novel 18F-labelled PET tracer targeting P2X7

et al. 2017

Self Cite

View full text Add to dashboard Cite

show abstract

“…[164,179] P2XRs are involved in an umber of relevantp harmacological events such as pain, inflammation, and neurotransmitterr elease. [180,181] In contrast to AR and P2YRs,t he pharmacology of the P2XRs is underdeveloped at the present time. [164] Ac rystal structure of zebrafish P2X 4 Ri nt he closed state and another one bearing a molecule of ATPa re available.…”

Section: Nucleotidesmentioning

confidence: 99%

The Role of the Phosphorus Atom in Drug Design

2019

View full text Add to dashboard Cite

Although the phosphorus atom is found in a variety of oxidation states, most of the phosphorus‐containing molecules of pharmacological importance possess phosphorus in the form of phosphonate or phosphinate functional groups, or in a major oxidation state as a phosphate group. The most common occurrence of phosphorus in drugs is either in prodrugs or in compounds for which the phosphorus atom plays a role in the biological activity, such as in modified nucleotides, in metabolically stable analogues of metabolites bearing phosphate groups, and as bioisosteric analogues of carboxyl groups.

show abstract

“…To understand the molecular mechanisms underlying opening and closing of P2RX2 through ATP binding, different amino acids have been mutated using site-directed mutagenesis in purified rat P2RX2 (rP2RX2) or human P2RX2 (hP2RX2) (Chataigneau et al, 2013 ; Jiang et al, 2013 ; Dal Ben et al, 2015 ; Habermacher et al, 2016 ). Mutations namely, F183C, T184C, and F289C causes 4–10-fold decrease in ATP binding to hP2X2 (Roberts et al, 2008 ; Chataigneau et al, 2013 ).…”

Section: Introductionmentioning

confidence: 99%

Characterization of ATPase Activity of P2RX2 Cation Channel

et al. 2016

View full text Add to dashboard Cite

P2X purinergic receptors are plasma membrane ATP-dependent cation channels that are broadly distributed in the mammalian tissues. P2RX2 is a modulator of auditory sensory hair cell mechanotransduction and plays an important role in hair cell tolerance to noise. In this study, we demonstrate for the first time in vitro and in cochlear neuroepithelium, that P2RX2 possesses the ATPase activity. We observed that the P2RX2 V60L human deafness mutation alters its ability to bind ATP, while the G353R has no effect on ATP binding or hydrolysis. A non-hydrolysable ATP assay using HEK293 cells suggests that ATP hydrolysis plays a significant role in the opening and gating of the P2RX2 ion channel. Moreover, the results of structural modeling of the molecule was in agreement with our experimental observations. These novel findings suggest the intrinsic ATPase activity of P2RX2 and provide molecular insights into the channel opening.

show abstract

Purinergic P2X receptors: Structural models and analysis of ligand-target interaction

Cited by 39 publications

References 101 publications

Design, synthesis and evaluation in an LPS rodent model of neuroinflammation of a novel 18F-labelled PET tracer targeting P2X7

Design, synthesis and evaluation in an LPS rodent model of neuroinflammation of a novel 18F-labelled PET tracer targeting P2X7

The Role of the Phosphorus Atom in Drug Design

Characterization of ATPase Activity of P2RX2 Cation Channel

Contact Info

Product

Resources

About