Pyrimidyn Compounds: Dual-Action Small Molecule Pyrimidine-Based Dynamin Inhibitors

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• 作者：Andrew B. McGeachie ; Luke R. Odell ; Annie Quan ; James A. Daniel ; Ngoc Chau ; Timothy A. Hill ; Nick N. Gorgani ; Damien J. Keating ; Michael A. Cousin ; Ellen M. van Dam ; Anna Mariana ; Ainslie Whiting ; Swetha Perera ; Aimee Novelle ; Kelly A. Young ; Fiona M. Deane ; Jayne Gilbert ; Jennette A. Sakoff ; Megan Chircop ; Adam McCluskey ; Phillip J. Robinson
• 刊名：ACS Chemical Biology
• 出版年：2013
• 出版时间：July 19, 2013
• 年：2013
• 卷：8
• 期：7
• 页码：1507-1518
• 全文大小：555K
• 年卷期：v.8,no.7(July 19, 2013)
• ISSN：1554-8937

文摘

Dynamin is required for clathrin-mediated endocytosis (CME). Its GTPase activity is stimulated by phospholipid binding to its PH domain, which induces helical oligomerization. We have designed a series of novel pyrimidine-based 鈥淧yrimidyn鈥?compounds that inhibit the lipid-stimulated GTPase activity of full length dynamin I and II with similar potency. The most potent analogue, Pyrimidyn 7, has an IC₅₀ of 1.1 渭M for dynamin I and 1.8 渭M for dynamin II, making it among the most potent dynamin inhibitors identified to date. We investigated the mechanism of action of the Pyrimidyn compounds in detail by examining the kinetics of Pyrimidyn 7 inhibition of dynamin. The compound competitively inhibits both GTP and phospholipid interactions with dynamin I. While both mechanisms of action have been previously observed separately, this is the first inhibitor series to incorporate both and thereby to target two distinct domains of dynamin. Pyrimidyn 6 and 7 reversibly inhibit CME of both transferrin and EGF in a number of non-neuronal cell lines as well as inhibiting synaptic vesicle endocytosis (SVE) in nerve terminals. Therefore, Pyrimidyn compounds block endocytosis by directly competing with GTP and lipid binding to dynamin, limiting both the recruitment of dynamin to membranes and its activation. This dual mode of action provides an important new tool for molecular dissection of dynamin鈥檚 role in endocytosis.