NF 449 is an efficient purine receptor antagonist, showing high selectivity for P2X1 (IC50 values for rP2X1, rP2X1+5, rP2X2+3, and rP2X3 receptors are 0.28, 0.69, 120, and 1820nM, respectively) [1]. NF 449 is a Gsα-selective G protein antagonist that inhibits the rate of GTP [γS] binding to Gsα-s, inhibits the stimulation of adenylate cyclase activity, and blocks the coupling of β-adrenergic receptors with Gs [2]. NF 449 can be used to distinguish the effects mediated by P2X1 and P2X7 in natural tissues [3].
In vitro, NF 449 (10μM; 5min) treatment significantly inhibited the activation of Gαs and cAMP production in GLP-1 (7-36) amide-treated muscle cells, and blocked the increase in PKA activity [4]. NF 449 (2μM; 10min) pretreatment of multi-cellular preparations of HGOA almost completely inhibited the contraction response caused by 10μM αβ-meATP, with the average normalized peak contraction amplitude changing from 100±12% to 6±2% [5].
In vivo, NF 449 (10, 50mg/kg/day; i.v.) single treatment inhibited the P2X1 receptor in systemic thromboembolic model WT mice and reduced intravascular platelet aggregation, but did not prolong the bleeding time. At higher doses, NF 449 inhibited ex vivo platelet aggregation, inhibited three types of platelet P2 receptors, and further reduced platelet consumption [6]. NF 449 (10mg/kg; twice; i.v.) treatment significantly reduced the protein content of BAL in LPS-sensitized H-2d BALB/c mice, and at the same time, the area of interstitial small artery periphery edema in the mice also decreased [7].
References:
[1] Rettinger J, et al. Profiling at recombinant homomeric and heteromeric rat P2X receptors identifies the suramin analogue NF 449 as a highly potent P2X1 receptor antagonist. Neuropharmacology. 2005;48(3):461-468.
[2] Hohenegger M, et al. Gsalpha-selective G protein antagonists. Proc Natl Acad Sci U S A. 1998;95(1):346-351.
[3] Hülsmann M, et al. NF 449, a novel picomolar potency antagonist at human P2X1 receptors. Eur J Pharmacol. 2003;470(1-2):1-7.
[4] May AT, et al. Identification of expression and function of the glucagon-like peptide-1 receptor in colonic smooth muscle. Peptides. 2019; 112:48-55.
[5] Nichols CM, et al. Vascular smooth muscle cells from small human omental arteries express P2X1 and P2X4 receptor subunits. Purinergic Signal. 2014;10(4):565-572.
[6] Hechler B, et al. Inhibition of platelet functions and thrombosis through selective or nonselective inhibition of the platelet P2 receptors with increasing doses of NF 449 [4,4',4'',4'''-(carbonylbis(imino-5,1,3-benzenetriylbis-(carbonylimino)))tetrakis-benzene-1,3-disulfonic acid octasodium salt]. J Pharmacol Exp Ther. 2005;314(1):232-243.
[7] El Mdawar M B, et al. The ATP-gated P2X1 ion channel contributes to the severity of antibody-mediated transfusion-related acute lung injury in mice[J]. Scientific Reports, 2019, 9(1): 5159.
NF 449是一种高效的嘌呤能受体拮抗剂，对P2X1表现出高选择性（对于rP2X1、rP2X1+5、rP2X2+3和rP2X3受体，IC50值分别为0.28、0.69、120、1820nM）[1]。NF 449是一种Gsα选择性G蛋白(G Protein)拮抗剂，抑制GTP[γS]与Gsα-s结合的速率，抑制腺苷酸环化酶活性的刺激，阻断β-肾上腺素能受体与Gs的偶联 [2]。NF 449可用于区分天然组织中P2X1和P2X7介导的作用 [3]。。
在体外，NF 449（10μM; 5min）处理显著抑制GLP-1 (7-36)酰胺处理的肌细胞Gαs的激活和cAMP的产生，阻断了PKA活性的增加 [4]。NF 449（2μM; 10min）预处理HGOA 多细胞样本几乎完全抑制了由10μM αβ-meATP引起的收缩反应，平均归一化峰收缩幅度由100±12%变为6±2% [5]。
在体内，NF 449（10, 50mg/kg/day; i.v.）单次治疗在低剂量下抑制了系统性血栓栓塞模型WT小鼠P2X1受体并降低血管内血小板聚集，但并未延长出血时间。而在较高剂量下，NF 449抑制离体血小板聚集，NF 449抑制了三种血小板P2受体，血小板消耗进一步减少 [6]。NF 449（10mg/kg/d; twice; i.v.）治疗显著降低了LPS致敏的H-2d BALB/c小鼠动物中BAL的蛋白表达，同时小鼠的间质性小动脉周围水肿面积也减小[7]。