权益分类	功能权益	普通用户	{{item.name}}会员
{{category.name}}	{{benefitItem.name}}

Design of Bifunctional Inhibitors for Enzymes in Coagulation-Fibrinolytic System

凝血纤溶系统双功能酶抑制剂的设计

基本信息

批准号：
08672573
负责人：
TSUDA Yuko
金额：
$ 1.15万
依托单位：
Kobe Gakuin University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
1996
资助国家：
日本
起止时间：
1996 至 1997
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-08672573/
关键词：
bifunctional inhibitor coagulation fibrinolysis

项目摘要

1. Evaluation of bifunctional thrombin inhibitors in vivo and further investigation of multifunctional thrombin inhibitors targeting thrombus : The antithrombotic effect of potent bifunctional thrombin inhibitors on He-Ne laser-induced thrombosis was evaluated in rat and compared with other types of thrombin inhibitors. The anthithrombotic activity of bifunctional inhibitors was higher than that of hirudin and hirulogs and comparable to that of argatroban, typical active site directed thrombin inhibitor. This result made us to design a multifunctional thrombin inhibitor targeting thrombus. Such a inhibitor would be not only more effective but also less side effects. A motif of Arg-Gly-Asp was introduced to the linker part on the molecule of bifunctional inhibitor and anthithrombin activities in vitro were examined.2. Design of bifunctional plasmin inhibitor : We design bifunctional plasmin inhibitor, which consists of an active site blocking sequence, t-AMCHA-Tyr-EACA-NH_2 (IC_<50>=4.6x10^<-4>M) , alysine binding sites (LBS) blocking moiety, Lys (IC_<50>=5.0x10^<-2>M) , and a linker, connecting those inhibitor moieties. Among the peptides synthesized, P-1 inhibited plasmin with a Ki value of 1.9x10^<-6>M in the amidolytic assay. The addition of t-AMCHA,a LBS binding inhibitor, reduced inhibitory activity of P-1 to a Ki value of 1.4x10^<-5>M.This result suggests that P-1 binds to the LBS of plasmin and acts as a bifunctional plasmin inhibitor.3. Design of plasma kallikrein (PK) inhibitor : The synthetic PK inhibitor, PKSI-527 consists of three parts. Each part was replaced by analogues in an attempt to improve the potency and the selectivity of PKSI-527. We found the peptide that inhibited PK with a high selectivity and an IC_<50> value in the same range of PKSI-527, This compound could be incorporated to bifunctional PK inhibitor as a active site blocking segment.

1.双功能凝血酶抑制剂的体内评价及靶向血栓的多功能凝血酶抑制剂的进一步研究：在大鼠体内评价了有效的双功能凝血酶抑制剂对He-Ne激光诱导血栓形成的抗血栓作用，并与其他类型的凝血酶抑制剂进行了比较。双功能抑制剂的抗血栓活性高于水蛭素和水蛭素，并与典型的活性位点定向凝血酶抑制剂阿加曲班相当。这一结果促使我们设计一种靶向血栓的多功能凝血酶抑制剂。这种抑制剂不仅更有效，而且副作用更小。在双功能抑制剂分子的连接子部分引入Arg-Gly-Asp基序，并检测其体外抗凝血酶活性.双功能纤溶酶抑制剂的设计：我们设计了双功能纤溶酶抑制剂，其由活性位点封闭序列t-AMCHA-Tyr-EACA-NH_2（IC_<50>= 4.6 × 10 ~ 4 μ <-4>M）、赖氨酸结合位点（LBS）封闭部分Lys（IC_<50>= 5.0 × 10 ~ 4 μ <-2>M）和连接这些抑制剂部分的接头组成。在合成的肽中，P-1在酰胺分解测定中抑制纤溶酶，Ki值为1.9x10^<-6>M。加入t-AMCHA（一种LBS结合抑制剂）后，P-1的抑制活性降低至Ki值为1.4 × 10 - 4 μ <-5>M。该结果表明P-1与纤溶酶的LBS结合，并作为双功能纤溶酶底物发挥作用。血浆激肽释放酶（PK）抑制剂的设计：合成的PK抑制剂PKSI-527由三部分组成。每个部分都用类似物代替，以提高PKSI-527的效力和选择性。我们发现该肽具有高选择性的PK抑制作用，其IC值<50>与PKSI-527相同，该化合物可作为活性位点阻断片段掺入双功能PK抑制剂中。