Characterization of human and rodent native and recombinant adenosine A2B receptors by radioligand binding studies

被引：21

作者：

Bertarelli D.C.G. ^{[1
]}

Diekmann M. ^{[1
]}

Hayallah A.M. ^{[1
]}

Rüsing D. ^{[2
]}

Iqbal J. ^{[1
]}

Preiss B. ^{[1
]}

Verspohl E.J. ^{[2
]}

Müller C.E. ^{[3
]}

机构：

[1] Pharmaceutical Institute, Pharmaceutical Sciences Bonn (PSB), University of Bonn, Bonn

[2] Institute of Pharmaceutical and Medicinal Chemistry, Department of Pharmacology, University of Münster, Münster

[3] Pharmazeutisches Institut, Pharmazeutische Chemie Poppelsdorf, Bonn 53115

来源：

Purinergic Signalling | 2006年 / 2卷 / 3期

关键词：

[!sup]3[!/sup]H]PSB-298; A[!sub]2B[!/sub] antagonist radioligand; Adenosine A[!sub]2B[!/sub] receptor; CHO cells; HEK-293; cells; INS-1; NG108-15; Sodium shift assays;

D O I：

10.1007/s11302-006-9012-4

中图分类号：

学科分类号：

摘要：

Adenosine A2B receptors of native human and rodent cell lines were investigated using [3H]PSB-298 [(8-{4-[2-(2-hydroxyethylamino)-2-oxoethoxy]phenyl}-1-propylxanthine] in radioligand binding studies. [3H]PSB-298 showed saturable and reversible binding. It exhibited a KD value of 60 ± 1 nM and limited capacity (Bmax = 3.511 fmol per milligram protein) at recombinant human adenosine A2B receptors expressed in human embryonic kidney cells (HEK-293). The addition of sodium chloride (100 mM) led to a threefold increase in the number of binding sites recognized by the radioligand. The curve of the agonist 5′-N-ethylcarboxamidoadenosine (NECA) was shifted to the right in the presence of NaCl, while the curve of the antagonist PSB-298 was shifted to the left, indicating that PSB-298 may be an inverse agonist at A2B receptors. Adenosine A2B receptors were shown to be the major adenosine A2 receptor subtype on the mouse neuroblastoma x rat glioma hybrid cell line NG108-15 cells. Binding studies at rat INS-1 cells (insulin secreting cell line) demonstrated that [3H]PSB-298 is a selective radioligand for adenosine A2B binding sites in this cell line. © Springer Science + Business Media B.V. 2006.

引用

页码：559 / 571

页数：12

共 40 条

[1]

Volpini R., Costanzi S., Vittori S., Et al., Medicinal chemistry and pharmacology of A2B adenosine receptors, Curr Top Med Chem, 3, pp. 427-433, (2003)

[2]

Holgate S.T., The identification of the adenosine A2B receptor as a novel therapeutic target in asthma, Br J Pharmacol, 145, pp. 1009-1015, (2005)

[3]

Abo-Salem O.M., Hayallah A.M., Bilkei-Gorzo A., Et al., Antinociceptive effects of novel A2B adenosine receptor antagonists, J Pharmacol Exp Ther, 308, pp. 358-366, (2004)

[4]

Feoktistov I., Polosa R., Holgate S.T., Et al., Adenosine A2B receptors: A novel therapeutic target in asthma?, Trends Pharmacol Sci, 19, pp. 148-153, (1998)

[5]

Harada H., Asano O., Hoshino Y., Et al., 2-Alkynyl-8-aryl-9-methyladenosines as novel adenosine receptor antagonists: Their synthesis and structure-activity relationship toward hepatic glucose production induced via agonism of the A2B receptor, J Med Chem, 44, pp. 170-179, (2001)

[6]

Ji X.D., Jacobson K.A., Use of the triazolotriazine [3 H]ZM 241385 as a radioligand at recombinant human A2B adenosine receptors, Drug Des Discov, 16, pp. 217-226, (1999)

[7]

Linden J., Thai T., Figler H., Et al., Characterization of human A2B adenosine receptors: Radioligand binding, western blotting, and coupling to Gq in human embryonic kidney 293 cells and HMC-1 mast cells, Mol Pharmacol, 56, pp. 705-713, (1999)

[8]

Robeva A.S., Woodard R., Jin X., Et al., Molecular characterization of recombinant human adenosine receptors, Drug Dev Res, 39, pp. 243-252, (1996)

[9]

Ji X.D., Kim Y.C., Ahern D.G., Et al., [3H]MRS 1754, a selective antagonist radioligand for A2B adenosine receptors, Biochem Pharmacol, 61, pp. 657-663, (2001)

[10]

Baraldi P.G., Aghazadeh Tabrizi M., Preti D., Et al., [3H]MRE 2029-F20, a selective antagonist radioligand for the human A2B adenosine receptors, Bioorg Med Chem, 14, pp. 3607-3610, (2004)

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