Amira Pharmaceuticals, San Diego, CA 92121, USA.
Br J Pharmacol. 2010 Aug;160(7):1699-713. doi: 10.1111/j.1476-5381.2010.00828.x.
The aim of this study was to assess the potential of an antagonist selective for the lysophosphatidic acid receptor, LPA(1), in treating lung fibrosis We evaluated the in vitro and in vivo pharmacological properties of the high affinity, selective, oral LPA(1)-antagonist (4'-{4-[(R)-1-(2-chloro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}-biphenyl-4-yl)-acetic acid (AM966).
The potency and selectivity of AM966 for LPA(1) receptors was determined in vitro by calcium flux and cell chemotaxis assays using recombinant and native cell cultures. The in vivo efficacy of AM966 to reduce tissue injury, vascular leakage, inflammation and fibrosis was assessed at several time points in the mouse bleomycin model.
AM966 was a potent antagonist of LPA(1) receptors, with selectivity for this receptor over the other LPA receptors. In vitro, AM966 inhibited LPA-stimulated intracellular calcium release (IC(50)= 17 nM) from Chinese hamster ovary cells stably expressing human LPA(1) receptors and inhibited LPA-induced chemotaxis (IC(50)= 181 nM) of human IMR-90 lung fibroblasts expressing LPA(1) receptors. AM966 demonstrated a good pharmacokinetic profile following oral dosing in mice. In the mouse, AM966 reduced lung injury, vascular leakage, inflammation and fibrosis at multiple time points following intratracheal bleomycin instillation. AM966 also decreased lactate dehydrogenase activity and tissue inhibitor of metalloproteinase-1, transforming growth factor beta1, hyaluronan and matrix metalloproteinase-7, in bronchoalveolar lavage fluid.
These findings demonstrate that AM966 is a potent, selective, orally bioavailable LPA(1) receptor antagonist that may be beneficial in treating lung injury and fibrosis, as well as other diseases that are characterized by pathological inflammation, oedema and fibrosis.
本研究旨在评估一种对溶血磷脂酸受体 LPA(1)具有选择性的拮抗剂在治疗肺纤维化方面的潜力。我们评估了高亲和力、选择性、口服 LPA(1)拮抗剂（4'-{4-[(R)-1-(2-氯-苯基）-乙氧羰基氨基]-3-甲基异恶唑-5-基}-联苯-4-基）-乙酸（AM966）的体外和体内药理学特性。
通过使用重组和天然细胞培养物的钙通量和细胞趋化性测定，在体外确定 AM966 对 LPA(1)受体的效力和选择性。在小鼠博莱霉素模型中，在多个时间点评估 AM966 降低组织损伤、血管渗漏、炎症和纤维化的体内疗效。
AM966 是 LPA(1)受体的有效拮抗剂，对该受体的选择性高于其他 LPA 受体。在体外，AM966 抑制人 LPA(1)受体稳定表达的中国仓鼠卵巢细胞中 LPA 刺激的细胞内钙释放（IC(50)=17 nM），并抑制人 IMR-90 肺成纤维细胞中 LPA(1)受体诱导的趋化性（IC(50)=181 nM）。AM966 在小鼠中口服给药后具有良好的药代动力学特征。在小鼠中，AM966 在经气管内博莱霉素滴注后多个时间点降低肺损伤、血管渗漏、炎症和纤维化。AM966 还降低了支气管肺泡灌洗液中的乳酸脱氢酶活性和组织金属蛋白酶抑制剂-1、转化生长因子β1、透明质酸和基质金属蛋白酶-7。
这些发现表明，AM966 是一种有效的、选择性的、口服生物利用的 LPA(1)受体拮抗剂，可能有益于治疗肺损伤和纤维化以及其他以病理性炎症、水肿和纤维化为特征的疾病。
