Buntinx Mieke, Hermans Bart, Goossens Jan, Moechars Dieder, Gilissen Ron A H J, Doyon Julien, Boeckx Staf, Coesemans Erwin, Van Lommen Guy, Van Wauwe Jean P
Johnson & Johnson Pharmaceutical Research and Development, Turnhoutseweg 30, B-2340 Beerse, Belgium.
J Pharmacol Exp Ther. 2008 Oct;327(1):1-9. doi: 10.1124/jpet.108.140723. Epub 2008 Jul 3.
The interaction between CC chemokine receptor 2 (CCR2) with monocyte chemoattractant proteins, such as MCP-1, regulates the activation and recruitment of inflammatory leukocytes. In this study, we characterized (S)-3-[3,4-difluoro-phenyl)-propyl]-5-isoxazol-5-yl-2-thioxo-2,3-dihydro-1H-imidazole-4-carboxyl acid methyl ester (JNJ-27141491) as a noncompetitive and orally active functional antagonist of human (h)CCR2. JNJ-27141491 strongly suppressed hCCR2-mediated in vitro functions, such as MCP-1-induced guanosine 5'-O-(3-[(35)S]thio)triphosphate binding; MCP-1, -3, and -4-induced Ca(2+) mobilization; and leukocyte chemotaxis toward MCP-1 (IC(50) = 7-97 nM), whereas it had little or no effect on the function of other chemokine receptors tested. The inhibition of CCR2 function was both insurmountable and reversible, consistent with a noncompetitive mode of action. JNJ-27141491 blocked the binding of (125)I-MCP-1 to human monocytes (IC(50) = 0.4 microM), but it failed to affect MCP-1 binding to mouse, rat, and dog cells (IC(50) > 10 microM). Therefore, transgenic mice, in which the mouse (m)CCR2 gene was replaced by the human counterpart, were generated for in vivo testing. In these mice, oral administration of JNJ-27141491 dose-dependently [5-40 mg/kg q.d. (once daily) or b.i.d.] inhibited monocyte and neutrophil recruitment to the alveolar space 48 h after intratracheal mMCP-1/lipopolysaccharide instillation. Furthermore, treatment with JNJ-27141491 (20 mg/kg q.d.) significantly delayed the onset and temporarily reduced neurological signs in an experimental autoimmune encephalomyelitis model of multiple sclerosis. Taken together, these results identify JNJ-27141491 as a noncompetitive, functional antagonist of hCCR2, capable of exerting oral anti-inflammatory activity in transgenic hCCR2-expressing mice.
C-C趋化因子受体2（CCR2）与单核细胞趋化蛋白（如MCP-1）之间的相互作用调节炎症白细胞的激活和募集。在本研究中，我们将（S）-3-[（3,4-二氟苯基）丙基]-5-异恶唑-5-基-2-硫代-2,3-二氢-1H-咪唑-4-羧酸甲酯（JNJ-27141491）鉴定为人类（h）CCR2的非竞争性口服活性功能拮抗剂。JNJ-27141491强烈抑制hCCR2介导的体外功能，如MCP-1诱导的鸟苷5'-O-（3-[(35)S]硫代）三磷酸结合；MCP-1、-3和-4诱导的Ca(2+)动员；以及白细胞对MCP-1的趋化作用（IC(50)=7-97 nM），而对所测试的其他趋化因子受体的功能几乎没有影响。CCR2功能的抑制是不可克服且可逆的，这与非竞争性作用模式一致。JNJ-27141491阻断(125)I-MCP-1与人单核细胞的结合（IC(50)=0.4 microM），但不影响MCP-1与小鼠、大鼠和犬细胞的结合（IC(50)>10 microM）。因此，构建了将小鼠（m）CCR2基因替换为人CCR2基因的转基因小鼠用于体内试验。在这些小鼠中，口服JNJ-27141491剂量依赖性地[5-40 mg/kg每日一次（q.d.）或每日两次（b.i.d.）]抑制气管内注入mMCP-1/脂多糖后48小时单核细胞和中性粒细胞向肺泡腔的募集。此外，在多发性硬化症的实验性自身免疫性脑脊髓炎模型中，用JNJ-27141491（20 mg/kg q.d.）治疗可显著延迟发病并暂时减轻神经症状。综上所述，这些结果表明JNJ-27141491是hCCR2的非竞争性功能拮抗剂，能够在表达转基因hCCR2的小鼠中发挥口服抗炎活性。
