Characterization of peripheral human cannabinoid receptor (hCB2) expression and pharmacology using a novel radioligand, [35S]Sch225336
Characterization of peripheral human cannabinoid receptor (hCB2) expression and pharmacology using a novel radioligand, [35S]Sch225336
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DOI：
10.1074/jbc.m602364200
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发表时间：
2006-09-22
影响因子：
4.8
通讯作者：
Hipkin, R. William
中科院分区：
文献类型：
--
作者：
Gonsiorek, Waldemar;Hesk, David;Hipkin, R. William
表征外周人大麻素受体（hCB 2）的内源性表达和药理学的研究受到缺乏真实的抗hCB 2抗体和缺乏具有CB 2选择性的放射性配体的阻碍。我们最近描述了一种新的CB 2反向激动剂，N-[1（S）-[4-[[4-甲氧基2-[（4-甲氧基苯基）磺酰基]苯基]磺酰基]苯基]乙基]甲烷磺酰胺（Sch 225336），其以高亲和力和相对于hCB 1的优异选择性结合hCB 2。制备Sch 225336的前体伯胺，并直接与[S-35]甲磺酰氯（由市售[S-35]甲磺酸合成）反应生成[S-35] Sch 225336。[35 S] Sch 225336具有高比活性（> 1400 Ci/mmol）和对hCB 2的亲和力（65 pM）。使用[S-35] Sch 225336，我们测定了造血细胞和细胞系，以定量hCB 2的表达和药理学。最后，我们使用[S-35] Sch 225336对淋巴组织中的CB 2进行了详细的放射自显影分析。基于这些数据，我们得出结论，[S-35] Sch 225336代表了用于研究血细胞和组织中内源性表达的CB 2的独特放射性配体。
Studies to characterize the endogenous expression and pharmacology of peripheral human cannabinoid receptor (hCB2) have been hampered by the dearth of authentic anti-hCB2 antibodies and the lack of radioligands with CB2 selectivity. We recently described a novel CB2 inverse agonist, N-[1(S)-[4-[[4- methoxy2-[(4-methoxyphenyl) sulfonyl] phenyl] sulfonyl] phenyl] ethyl] methane-sulfonamide (Sch225336), that binds hCB2 with high affinity and excellent selectivity versus hCB1. The precursor primary amine of Sch225336 was prepared and reacted directly with [S-35] mesyl chloride ( synthesized from commercially obtained [S-35] methane sulfonic acid) to generate [S-35] Sch225336. [ 35S] Sch225336 has high specific activity (> 1400 Ci/mmol) and affinity for hCB2( 65 pM). Using [S-35] Sch225336, we assayed hemopoietic cells and cell lines to quantitate the expression and pharmacology of hCB2. Lastly, we used [S-35] Sch225336 for detailed autoradiographic analysis of CB2 in lymphoid tissues. Based on these data, we conclude that [S-35] Sch225336 represents a unique radioligand for the study of CB2 endogenously expressed in blood cells and tissues.