title: Building a better dynasore: the dyngo compounds potently inhibit dynamin and endocytosis.
abstract: Dynamin GTPase activity increases when it oligomerizes either into helices in the presence of lipid templates or into rings in the presence of SH3 domain proteins. Dynasore is a dynamin inhibitor of moderate potency (IC₅₀ ~ 15 μM in vitro). We show that dynasore binds stoichiometrically to detergents used for in vitro drug screening, drastically reducing its potency (IC₅₀ = 479 μM) and research tool utility. We synthesized a focused set of dihydroxyl and trihydroxyl dynasore analogs called the Dyngo™ compounds, five of which had improved potency, reduced detergent binding and reduced cytotoxicity, conferred by changes in the position and/or number of hydroxyl substituents. The Dyngo compound 4a was the most potent compound, exhibiting a 37-fold improvement in potency over dynasore for liposome-stimulated helical dynamin activity. In contrast, while dynasore about equally inhibited dynamin assembled in its helical or ring states, 4a and 6a exhibited >36-fold reduced activity against rings, suggesting that they can discriminate between helical or ring oligomerization states. 4a and 6a inhibited dynamin-dependent endocytosis of transferrin in multiple cell types (IC₅₀ of 5.7 and 5.8 μM, respectively), at least sixfold more potently than dynasore, but had no effect on dynamin-independent endocytosis of cholera toxin. 4a also reduced synaptic vesicle endocytosis and activity-dependent bulk endocytosis in cultured neurons and synaptosomes. Overall, 4a and 6a are improved and versatile helical dynamin and endocytosis inhibitors in terms of potency, non-specific binding and cytotoxicity. The data further suggest that the ring oligomerization state of dynamin is not required for clathrin-mediated endocytosis.
authors.affiliation: Chemistry, Centre for Chemical Biology, School of Environmental and Life Sciences, The University of Newcastle, Callaghan, NSW, 2308, Australia.
authors.forename: Adam
authors.initials: A
authors.lastname: McCluskey
authors.forename: James A
authors.initials: JA
authors.lastname: Daniel
authors.forename: Gordana
authors.initials: G
authors.lastname: Hadzic
authors.forename: Ngoc
authors.initials: N
authors.lastname: Chau
authors.forename: Emma L
authors.initials: EL
authors.lastname: Clayton
authors.forename: Anna
authors.initials: A
authors.lastname: Mariana
authors.forename: Ainslie
authors.initials: A
authors.lastname: Whiting
authors.forename: Nick N
authors.initials: NN
authors.lastname: Gorgani
authors.forename: Jonathan
authors.initials: J
authors.lastname: Lloyd
authors.forename: Annie
authors.initials: A
authors.lastname: Quan
authors.forename: Lia
authors.initials: L
authors.lastname: Moshkanbaryans
authors.forename: Sai
authors.initials: S
authors.lastname: Krishnan
authors.forename: Swetha
authors.initials: S
authors.lastname: Perera
authors.forename: Megan
authors.initials: M
authors.lastname: Chircop
authors.forename: Lisa
authors.initials: L
authors.lastname: von Kleist
authors.forename: Andrew B
authors.initials: AB
authors.lastname: McGeachie
authors.forename: Mark T
authors.initials: MT
authors.lastname: Howes
authors.forename: Robert G
authors.initials: RG
authors.lastname: Parton
authors.forename: Michael
authors.initials: M
authors.lastname: Campbell
authors.forename: Jennette A
authors.initials: JA
authors.lastname: Sakoff
authors.forename: Xuefeng
authors.initials: X
authors.lastname: Wang
authors.forename: Jian-Yuan
authors.initials: JY
authors.lastname: Sun
authors.forename: Mark J
authors.initials: MJ
authors.lastname: Robertson
authors.forename: Fiona M
authors.initials: FM
authors.lastname: Deane
authors.forename: Tam H
authors.initials: TH
authors.lastname: Nguyen
authors.forename: Frederic A
authors.initials: FA
authors.lastname: Meunier
authors.forename: Michael A
authors.initials: MA
authors.lastname: Cousin
authors.forename: Phillip J
authors.initials: PJ
authors.lastname: Robinson
mesh_terms: D000818:Animals; D045744:Cell Line, Tumor; D002478:Cells, Cultured; D002772:Cholera Toxin / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D055808:Drug Discovery; D034281:Dynamins / Q000037:antagonists & inhibitors* / Q000378:metabolism; D004705:Endocytosis / Q000187:drug effects*; D057166:High-Throughput Screening Assays; D006801:Humans; D006835:Hydrazones / Q000138:chemical synthesis / Q000737:chemistry / Q000494:pharmacology*; D009284:Naphthols / Q000737:chemistry / Q000494:pharmacology*; D009474:Neurons / Q000187:drug effects / Q000378:metabolism; D011485:Protein Binding; D021381:Protein Transport; D051381:Rats; D017207:Rats, Sprague-Dawley; D012756:Sheep; D013572:Synaptic Vesicles / Q000187:drug effects / Q000378:metabolism; D061250:Transferrins / Q000378:metabolism
chemical_list: D006835:Hydrazones; C511794:N'-(3,4-dihydroxybenzylidene)-3-hydroxy-2-naphthahydrazide; D009284:Naphthols; D061250:Transferrins; C568360:dyngo-4a; D002772:Cholera Toxin; D034281:Dynamins
keywords: bulk endocytosis; drug discovery; dynamin; high-throughput screening; small-molecule inhibitors; synaptic vesicle endocytosis
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