Molecular cloning and radioligand binding characterization of the chemokine receptor CCR5 from rhesus macaque and human
Molecular cloning and radioligand binding characterization of the chemokine receptor CCR5 from rhesus macaque and human
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DOI：
10.1016/j.bcp.2005.10.024
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发表时间：
2005-12-19
影响因子：
5.8
通讯作者：
Holbrook, M
中科院分区：
文献类型：
--
作者：
Napier, C;Sale, H;Holbrook, M
本研究的目的是确定猕猴是否是新型CCR 5拮抗剂（如马拉韦罗（UK-427，857））临床前评价的合适种属。为此，我们从恒河猴中克隆并表达了CCR 5，并比较了[I-125] -MIP-1 β和[H-3]-马拉韦罗与人重组CCR 5的结合特性。[I-125]-MIP-1 β与猕猴（Kd = 0.24 +/- 0.05 nM）和人（Kd = 0.23 +/- 0.05 nM）CCR 5的结合具有相似的高亲和力，并且具有相似的动力学特性。在竞争结合研究中，一系列人趋化因子对猕猴CCR 5的亲和力也与人CCR 5相似。马拉韦罗抑制[I-125]-MIP-1 β与猕猴和人CCR 5结合的效力相似（IC 50 = 17.50 +/- 1.24 nM和7.18 +/- 0.93 nM，分别）和拮抗MIP-1 β诱导的猕猴和人的CCR 5介导的细胞内钙释放，具有相似的效力（IC 50分别为17.50 +/- 3.30 nM和12.07 +/- 1.89）。[3 H]maraviroc与猕猴（Kd = 1.36 +/- 0.07 nM）和人（Kd = 0.86 +/- 0.08 nM）的CCR 5具有高亲和力结合，但发现与猕猴CCR 5的解离速度快10倍。然而，与人受体一样，马拉韦罗被证明是猕猴CCR 5的高亲和力、强效功能性拮抗剂，因此表明猕猴应该是评价新型CCR 5拮抗剂的药理学、安全性和潜在机制相关毒理学的合适种属。(c)2005年爱思唯尔公司All rights reserved.
The aim of this study was to determine if macaque represents a suitable species for the preclinical evaluation of novel CCR5 antagonists, such as maraviroc (UK-427,857). To do this we cloned and expressed CCR5 from rhesus macaque and compared the binding properties of [I-125] -MIP-1 beta and [H-3]-maraviroc with human recombinant CCR5. [I-125]-MIP-1 beta bound with similar high affinity to CCR5 from macaque (K-d = 0.24 +/- 0.05 nM) and human (K-d = 0.23 +/- 0.05 nM) and with similar kinetic properties. In competition binding studies the affinity of a range of human chemokines for macaque CCR5 was also similar to human CCR5. Maraviroc inhibited binding of [I-125]-MIP-1 beta to CCR5 from macaque and human with similar potency (IC50 = 17.50 +/- 1.24 nM and 7.18 +/- 0.93 nM, respectively) and antagonised MIP-1 beta induced intracellular calcium release mediated through CCR5 from macaque and human with similar potency (IC50 = 17.50 +/- 3.30 nM and 12.07 +/- 1.89, respectively). [3 H]maraviroc bound with high affinity to CCR5 from macaque (Kd = 1.36 +/- 0.07 nM) and human (Kd = 0.86 +/- 0.08 nM), but was found to dissociate similar to 10-fold more quickly from macaque CCR5. However, as with the human receptor, maraviroc was shown to be a high affinity, potent functional antagonist of macaque CCR5 thereby indicating that the macaque should be a suitable species in which to evaluate the pharmacology, safety and potential mechanism-related toxicology of novel CCR5 antagonists. (c) 2005 Elsevier Inc. All rights reserved.