A challenge finding P2X1 and P2X4 ligands

Beswick, Paul J.; Wahab, Ben; Honey, Mark A.; Paradowski, Michael; Jiang, Ke; Lochner, Martin; Murrell‐Lagnado, Ruth D.; Thompson, Andrew

doi:10.1016/j.neuropharm.2019.107674

Cited by 7 publications

(2 citation statements)

References 30 publications

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“…In both cases molecular models of human P2X7 based on homologous P2X receptor crystal structures were used and hit compounds with micromolar potency were obtained. This previous success with P2X7 models indicates that our strategy is valid, and a similar approach to ours has recently been carried out on a molecular model of human P2X4 ( Beswick et al, 2019 ), discovering hit compounds, but with very low (high micromolar) potency. There has been a lower success with a similar approach for human P2X4 and the discovery of antagonists using the hP2X4 orthosteric pocket in virtual screenings has been more challenging.…”

Section: Discussionmentioning

confidence: 58%

“…Structure-based screening on the ATP-binding (orthosteric) pocket of P2X7 has also been used successfully, leading to the identification of hit compounds with selectivity for P2X7 over P2X4 ( Caseley et al, 2016 ). A similar approach has also been used to discover P2X4 and P2X1 antagonists, although their potency was in the high micromolar range ( Beswick et al, 2019 ).…”

Section: Introductionmentioning

confidence: 99%

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Identification of a novel P2X7 antagonist using structure-based virtual screening

et al. 2023

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P2X4 and P2X7 receptors are ATP-gated ion channels, which play important roles in neuropathic and inflammatory pain, and as such they are important drug targets in diseases of inflammatory origin. While several compounds targeting P2X4 and P2X7 receptors have been developed using traditional high-throughput screening approaches, relatively few compounds have been developed using structure-based design. We initially set out to develop compounds targeting human P2X4, by performing virtual screening on the orthosteric (ATP-binding) pocket of a molecular model of human P2X4 based on the crystal structure of the Danio rerio receptor. The screening of a library of approximately 300,000 commercially available drug-like compounds led to the initial selection of 17 compounds; however, none of these compounds displayed a significant antagonist effect at P2X4 in a Fluo-4 ATP-induced calcium influx assay. When the same set of compounds was tested against human P2X7 in an ATP-stimulated Yo-Pro1 dye uptake assay, one compound (an indeno(1,2-b)pyridine derivative; GP-25) reduced the response by greater than 50% when applied at a concentration of 30 µM. GP-25 displayed an IC50 value of 8.7 μM at human P2X7 and 24.4 μM at rat P2X7, and was confirmed to be active using whole-cell patch clamp electrophysiology and not cytotoxic. Schild analysis suggested that mode of action of GP-25 was orthosteric. Screening of a further 16 commercially available analogues of GP-25 led to the discovery of five additional compounds with antagonist activity at human P2X7, enabling us to investigate the structure-activity relationship. Finally, docking of the R- and S-enantiomers of GP-25 into the orthosteric pocket of molecular models of human P2X4 and human P2X7 revealed that, while both enantiomers were able to make multiple interactions between their carboxyl moieties and conserved positively charged amino-acids in human P2X7, only the S-enantiomer of GP-25 was able to do this in human P2X4, potentially explaining the lack of activity of GP-25 at this receptor.

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Section: Discussionmentioning

confidence: 58%

Section: Introductionmentioning

confidence: 99%

Identification of a novel P2X7 antagonist using structure-based virtual screening

et al. 2023

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Development of Purinergic Receptor Agonists and Antagonists

Jacobson

2023

Purinergic Signaling in Neurodevelopment, Neuroinflammation and Neurodegeneration

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The P2X1 receptor as a therapeutic target

Bennetts

Mobbs

Ventura

et al. 2022

Purinergic Signalling

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Within the family of purinergic receptors, the P2X1 receptor is a ligand-gated ion channel that plays a role in urogenital, immune and cardiovascular function. Specifically, the P2X1 receptor has been implicated in controlling smooth muscle contractions of the vas deferens and therefore has emerged as an exciting drug target for male contraception. In addition, the P2X1 receptor contributes to smooth muscle contractions of the bladder and is a target to treat bladder dysfunction. Finally, platelets and neutrophils have populations of P2X1 receptors that could be targeted for thrombosis and inflammatory conditions. Drugs that specifically target the P2X1 receptor have been challenging to develop, and only recently have small molecule antagonists of the P2X1 receptor been available. However, these ligands need further biological validation for appropriate selectivity and drug-like properties before they will be suitable for use in preclinical models of disease. Although the atomic structure of the P2X1 receptor has yet to be determined, the recent discovery of several other P2X receptor structures and improvements in the field of structural biology suggests that this is now a distinct possibility. Such efforts may significantly improve drug discovery efforts at the P2X1 receptor.

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A challenge finding P2X1 and P2X4 ligands

Cited by 7 publications

References 30 publications

Identification of a novel P2X7 antagonist using structure-based virtual screening

Identification of a novel P2X7 antagonist using structure-based virtual screening

Development of Purinergic Receptor Agonists and Antagonists

The P2X1 receptor as a therapeutic target

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