Smart D, Sabido-David C, Brough S J, Jewitt F, Johns A, Porter R A, Jerman J C
Neuroscience Research, SmithKline Beecham Pharmaceuticals, New Frontiers Science Park, Third Avenue, Harlow, Essex CM19 5AW.
Br J Pharmacol. 2001 Mar;132(6):1179-82. doi: 10.1038/sj.bjp.0703953.
The pharmacology of various peptide and non-peptide ligands was studied in Chinese hamster ovary (CHO) cells stably expressing human orexin-1 (OX(1)) or orexin-2 (OX(2)) receptors by measuring intracellular calcium (Ca(2+)) using Fluo-3AM. Orexin-A and orexin-B increased Ca(2+) in CHO-OX(1) (pEC(50)=8.38+/-0.04 and 7.26+/-0.05 respectively, n=12) and CHO-OX(2) (pEC(50)=8.20+/-0.03 and 8.26+/-0.04 respectively, n=8) cells. However, neuropeptide Y and secretin (10 pM - 10 microM) displayed neither agonist nor antagonist properties in either cell-line. SB-334867-A (1-(2-Methyylbenzoxanzol-6-yl)-3-[1,5]naphthyridin-4-yl-urea hydrochloride) inhibited the orexin-A (10 nM) and orexin-B (100 nM)-induced calcium responses (pK(B)=7.27+/-0.04 and 7.23+/-0.03 respectively, n=8), but had no effect on the UTP (3 microM)-induced calcium response in CHO-OX(1) cells. SB-334867-A (10 microM) also inhibited OX(2) mediated calcium responses (32.7+/-1.9% versus orexin-A). SB-334867-A was devoid of agonist properties in either cell-line. In conclusion, SB-334867-A is a non-peptide OX(1) selective receptor antagonist.
通过使用Fluo-3AM测量细胞内钙浓度（Ca(2+)），研究了各种肽类和非肽类配体在中国仓鼠卵巢（CHO）细胞中的药理学特性，这些细胞稳定表达人食欲素-1（OX(1)）或食欲素-2（OX(2)）受体。食欲素-A和食欲素-B可增加CHO-OX(1)细胞（pEC(50)分别为8.38±0.04和7.26±0.05，n = 12）和CHO-OX(2)细胞（pEC(50)分别为8.20±0.03和8.26±0.04，n = 8）中的Ca(2+)。然而，神经肽Y和促胰液素（10 pM - 10 microM）在这两种细胞系中既不表现出激动剂特性，也不表现出拮抗剂特性。SB-334867-A（1-(2-甲基苯并恶唑-6-基)-3-[1,5]萘啶-4-基-脲盐酸盐）可抑制食欲素-A（10 nM）和食欲素-B（100 nM）诱导的钙反应（pK(B)分别为7.27±0.04和7.23±0.03，n = 8），但对CHO-OX(1)细胞中UTP（3 microM）诱导的钙反应没有影响。SB-334867-A（10 microM）也可抑制OX(2)介导的钙反应（与食欲素-A相比为32.7±1.9%）。SB-334867-A在这两种细胞系中均无激动剂特性。总之，SB-334867-A是一种非肽类OX(1)选择性受体拮抗剂。
