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        New Insights in Purinergic Therapy: Novel Antagonists for Uridine 5′-Triphosphate-Activated P2Y Receptors from Brazilian Flora

        Natiele Carla da Silva Ferreira,Roˆmulo Jose´ Soares-Bezerra,Rebeca Ferreira Couto da Silveira,Clayton Menezes da Silva,Carla Santos de Oliveira,Andrea Surrage Calheiros,Taˆnia Maria Alves,Carlos Leom 한국식품영양과학회 2019 Journal of medicinal food Vol.22 No.2

        P2Y2 and P2Y4 receptors are physiologically activated by uridine 5′-triphosphate (UTP) and are widely expressed in many cell types in humans. P2Y2 plays an important role in inflammation and proliferation of tumor cells, which could be attenuated with the use of antagonists. However, little is known about the physiological functions related to P2Y4, due to the lack of selective ligands for these receptors. This can be solved through the search for novel compounds with antagonistic activity. The aim of this study was to discover new potential antagonist candidates for P2Y2 and P2Y4 receptors from natural products. We applied a calcium measurement methodology to identify new antagonist candidates for these receptors. First, we established optimal conditions for the calcium assay using J774.G8, a murine macrophage cell line, which expresses functional P2Y2 and P2Y4 receptors and then, we performed the screening of plant extracts at a cutoff concentration of 50 μg/mL. ATP and ionomycin, known intracellular calcium inductors, were used to stimulate cells. The calculated EC50 were 11 μM and 103 nM, respectively. These cells also responded to the UTP stimulation with an EC50 of 1.021 μM. Screening assays were performed and a total of 100 extracts from Brazilian plants were tested. Joannesia princeps Vell. (stem) and Peixotoa A. Juss (flower and leaf) extracts stood out due to their ability to inhibit UTP-induced responses without causing cytotoxicity, and presented an IC50 of 32.32, 14.99, and 12.98 μg/mL, respectively. Collectively, our results point to the discovery of potential antagonist candidates from Brazilian flora for UTP-activated receptors.

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