Screening and X-ray Crystal Structure-based Optimization of Autotaxin (ENPP2) Inhibitors, Using a Newly Developed Fluorescence Probe
Screening and X-ray Crystal Structure-based Optimization of Autotaxin (ENPP2) Inhibitors, Using a Newly Developed Fluorescence Probe
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DOI：
10.1021/cb400150c
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发表时间：
2013-08-01
影响因子：
4
通讯作者：
Nagano, Tetsuo
中科院分区：
文献类型：
--
作者：
Kawaguchi, Mitsuyasu;Okabe, Takayoshi;Nagano, Tetsuo
自分泌运动因子（ATX），也称为外核苷酸焦磷酸酶/磷酸二酯酶2（ENPP2），最初被鉴定为一种肿瘤细胞自分泌运动因子，并被发现与血浆溶血磷脂酶D相同，后者是溶血磷脂产生溶血磷脂酸（LPA）的主要贡献者。因此，ATX被认为可调节LPA的生理和病理作用，包括血管生成、淋巴细胞运输、组织纤维化以及癌细胞侵袭和转移。因此，它是一个潜在的治疗靶点。在此，我们首先开发了一种灵敏且特异的ATX荧光探针TG - mTMP，并使用它在一个大型化学文库中筛选ATX抑制剂。这种探针在灵敏度或特异性方面优于先前可用的探针FS - 3和CPF4，使我们能够鉴定出几种新型的ATX抑制剂支架结构。我们以高分辨率（1.75 - 1.95埃）解析了ATX与命中化合物的复合物晶体结构，并利用这些信息指导对一种选定抑制剂的结构优化。优化后的化合物3BoA及其衍生物在体外和体内均表现出强大的ATX抑制活性。这些抑制剂有望成为了解ATX在体外和体内作用的有用工具，也可能是抗ATX治疗药物的候选者。
Autotaxin (ATX), also known as ectonucleotide pyrophosphatase/phosphodiesterase 2 (ENPP2), was originally identified as a tumor cell autocrine motility factor and was found to be identical to plasma lysophospholipase D, which is the predominant contributor to lysophosphatidic acid (LPA) production from lysophospholipids. ATX is therefore considered to regulate the physiological and pathological roles of LPA, including angiogenesis, lymphocyte trafficking, tissue fibrosis, and cancer cell invasion and metastasis. Thus, it is a potential therapeutic target. Here, we first developed a sensitive and specific ATX fluorescence probe, TG-mTMP, and used it to screen ATX inhibitors in a large chemical library. This probe, which is superior to previously available probes FS-3 and CPF4 in terms of sensitivity or specificity, enabled us to identify several novel ATX inhibitor scaffolds. We solved the crystal structures of ATX complexes with the hit compounds at high resolution (1.75-1.95 angstrom) and used this information to guide optimization of the structure of a selected inhibitor. The optimized compounds, 3BoA and its derivatives, exhibited potent ATX-inhibitory activity both in vitro and in vivo. These inhibitors are expected to be useful tools to understand the roles of ATX in vitro and in vivo and may also be candidate anti-ATX therapeutic agents.