Characterization of hepatocyte growth factor activator inhibitors which are being developed for a medicine

正在开发的药物肝细胞生长因子激活剂抑制剂的表征

基本信息

批准号：
13557012
负责人：
KITAMURA Naomi
金额：
$ 8.9万
依托单位：
TOKYO INSTITUTE OF TECHNOLOGY
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
2001
资助国家：
日本
起止时间：
2001 至 2002
项目状态：
已结题

项目摘要

It is reported that overaction of hepatocyte growth factor (HGF) causes kidney injury. Thus, it is required to develop a medicine which suppresses overaction of HGF. We identified two novel protease inhibitors (HAI-1 and HAI-2) which inhibit the activity of HGF activator. In this study, we examined whether these inhibitors could function as a medicine for diseases caused by overaction of HGF, and obtained the following results.1. HAI-1 has two Kunitz domains which are thought to be functional domains for the protease inhibitory activity of HAI-1. To determine the roles of the Kunitz domains in the inhibitory activity of HAI-1 against HGF activator, we constructed various human HAI-1 mutant proteins and examined their inhibitory activity against HGF activator. The N-terminal Kunitz domain had potent inhibitory activity, whereas the C-terminal Kunitz domain had only very weak activity. HAI-2 also has two Kunitz domains. We also examined the roles of Kunitz domains in the inhibitory activity of mouse HAI-2 against HGF activator. Unlike human HAI-1, the C-terminal Kunitz domain had potent inhibitory activity, whereas the N-terminal Kunitz domain had less activity. Therefore, it is necessary to examine which Kunitz domain of HAI-1 and HAI-2 is suitable for a medicine.2. To investigate whether the Kunitz domain of HAI suppresses activation of HGF in vivo, it is required to establish an assay system using experimental animals. We examined whether injection of HGF activator into rats induces activation of HGF. Injection of HGF activator induced activation of HGF in the liver of partially hepatectomized rats. This injection also induced liver regeneration. Therefore, this system would be useful to assay the inhibitory activity of the Kunitz domain of HAI.

据报道，肝细胞生长因子（HGF）的过度会导致肾脏损伤。因此，需要开发一种抑制HGF过度的药物。我们确定了两个新型蛋白酶抑制剂（HAI-1和HAI-2），它们抑制了HGF激活剂的活性。在这项研究中，我们检查了这些抑制剂是否可以充当HGF过度引起的疾病的药物，并获得了以下结果。1。 HAI-1具有两个Kunitz结构域，这些结构域被认为是HAI-1蛋白酶抑制活性的功能域。为了确定Kunitz结构域在HAI-1对HGF激活剂的抑制活性中的作用，我们构建了各种人类HAI-HAI-1突变蛋白，并检查了它们对HGF激活剂的抑制活性。 N末端Kunitz结构域具有有效的抑制活性，而C末端Kunitz结构域的活性仅非常弱。 HAI-2也有两个Kunitz域。我们还检查了库尼兹域在小鼠HAI-2对HGF激活剂的抑制活性中的作用。与人HAI-1不同，C端Kunitz结构域具有有效的抑制活性，而N端Kunitz结构域的活性较小。因此，有必要检查HAI-1和HAI-2的Kunitz领域适合药物2。为了研究HAI的Kunitz结构域是否抑制了HGF在体内的激活，需要使用实验动物建立一个测定系统。我们检查了将HGF激活剂注射到大鼠中是否诱导HGF的激活。注射HGF激活剂在部分肝切除大鼠的肝脏中诱导HGF激活。该注射还诱导肝脏再生。因此，该系统对于测定HAI Kunitz域的抑制活性将是有用的。