以组织蛋白酶L为靶位的骨质疏松新药筛选模型的研究  

A novel model for screening new drug of osteoporosis targeting on cathepsin L

作　　者：杨隽[1] 刘斌[1] 张月琴[1] 

机构地区：[1]中国协和医科大学中国医学科学院医药生物技术研究所,北京100050

出　　处：《中国抗生素杂志》2002年第5期264-266,312,共4页Chinese Journal of Antibiotics

摘　　要：对骨质疏松症分子水平上的研究发现 ,破骨细胞溶酶体向骨小腔分泌组织蛋白酶L ,进而造成以I 胶原为主要成分的骨支持蛋白分解亢进 ,成为导致骨质疏松症形成的重要因素之一。因此 ,寻找一种针对组织蛋白酶L的专一性抑制剂尤为重要。我们以组织蛋白酶L为靶位建立了寻找专一性抑制剂筛选模型。初筛模型 :以木瓜蛋白酶为靶酶筛选出半胱胺酸蛋白酶B、L的半数抑制浓度 (IC50 )之比 (B/L)为参照系数 ,以Chymostatin为强选择性对照 (B/L >30 ) ,以Leupeptin为弱选择性对照 (B/L <10 ) ,筛选出组织蛋白酶L的专一性抑制剂。应用此模型筛选得到JF 2 6 2、JF 2 87两株阳性菌。其发酵液对L型组织蛋白酶有强烈的选择性抑制活性。经初步鉴定 ,其产生菌培养特征及生理生化特性与文献报道的几株组织蛋白酶L抑制剂产生菌不同。Studies on molecular pharmacology proved that cathepsin L is a main proteinase responsible for bone resorption . When lysosome of osteoclast secrets precathepsin L to Howship's lacunae, it is auto-selfsplitted in pH<5 surroundings and then induce the hyper-decomposition of osseous I-collagen. So a specific inhibitor on cathepsin L which delayed the process of bone collagenolysis can be used in osteoporosis treatment.A screening model of specific inhibitor on cathepsin L was set up. The screening system consists of two parts: primarily screen out inhibitor on cys-proteinase by papain, then to screen out specific inhibitor on cathepsin L. It is referenced by M=IC 50 (Cathepsin B)/IC 50 (Papain), using chymostatin as a strong selective control and leupeptin as a weak selective control.Two strains(JF-262, JF-287) were screened out by this model. The crude extraction of zymotic fluid showed stronger selectivity than reported strains (FA-70).

关 键 词：骨质疏松症 组织蛋白酶L 筛选模型 

分 类 号：R965.1[医药卫生—药理学]