6-Furopyridine Hexamethylene Amiloride Is a Non-Selective P2X7 Receptor Antagonist
Abstract
:1. Introduction
2. Materials and Methods
2.1. Materials
2.2. Cell Lines
2.3. Expression Constructs
2.4. Transfection
2.5. Plate-Based Flow Cytometric Dye Uptake for Compound Screening
2.6. Flow Cytometric Uptake Assays for Characterization
2.7. Fura-2AM Ca2+ Response Assay
2.8. Whole Blood IL-1β Release Assay
2.9. CD23 Shedding Assay
2.10. Statistical Analysis
3. Results
3.1. 6-FPHMA Is a Novel P2X7 Antagonist
3.2. 6-FPHMA, but Not 6-FPA, Inhibits hP2X7-Mediated Dye Uptake
3.3. 6-FPHMA Inhibits hP2X7-Mediated Ca2+ Responses
3.4. 6-FPHMA Inhibits hP2X7-Mediated Dye Uptake in Human T Cells
3.5. 6-FPHMA Inhibits hP2X7-Mediated IL-1β Release and CD23 Shedding from Human Leukocytes
3.6. 6-FPHMA Inhibits mP2X7- and cP2X7-Mediated Dye Uptake
3.7. 6-FPHMA Inhibits hP2X4- and hP2Y-Mediated Ca2+ Responses
4. Discussion
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Cuthbertson, P.; Elhage, A.; Al-Rifai, D.; Sophocleous, R.A.; Turner, R.J.; Aboelela, A.; Majed, H.; Bujaroski, R.S.; Jalilian, I.; Kelso, M.J.; et al. 6-Furopyridine Hexamethylene Amiloride Is a Non-Selective P2X7 Receptor Antagonist. Biomolecules 2022, 12, 1309. https://doi.org/10.3390/biom12091309
Cuthbertson P, Elhage A, Al-Rifai D, Sophocleous RA, Turner RJ, Aboelela A, Majed H, Bujaroski RS, Jalilian I, Kelso MJ, et al. 6-Furopyridine Hexamethylene Amiloride Is a Non-Selective P2X7 Receptor Antagonist. Biomolecules. 2022; 12(9):1309. https://doi.org/10.3390/biom12091309
Chicago/Turabian StyleCuthbertson, Peter, Amal Elhage, Dena Al-Rifai, Reece A. Sophocleous, Ross J. Turner, Ashraf Aboelela, Hiwa Majed, Richard S. Bujaroski, Iman Jalilian, Michael J. Kelso, and et al. 2022. "6-Furopyridine Hexamethylene Amiloride Is a Non-Selective P2X7 Receptor Antagonist" Biomolecules 12, no. 9: 1309. https://doi.org/10.3390/biom12091309