title: Tubulin-based antimitotic mechanism of E7974, a novel analogue of the marine sponge natural product hemiasterlin.
abstract: E7974 is a synthetic analogue of the marine sponge natural product hemiasterlin. Here, we show that E7974, such as parental hemiasterlin, acts via a tubulin-based antimitotic mechanism. E7974 inhibits polymerization of purified tubulin in vitro with IC(50) values similar to those of vinblastine. In cultured human cancer cells, E7974 induces G(2)-M arrest and marked disruption of mitotic spindle formation characteristic of tubulin-targeted anticancer drugs. Extensive hypodiploid cell populations are seen in E7974-treated cells, indicating initiation of apoptosis after prolonged G(2)-M blockage. Consistent with this observation, E7974 induces caspase-3 activation and poly ADP ribose polymerase cleavage, typical biochemical markers of apoptosis. Only a short cellular exposure to E7974 is sufficient to induce maximum mitotic arrest, suggesting that E7974's antitumor effects in vivo may persist even after blood levels of the drug decrease after drug administration. Interactions of E7974 with purified tubulin were investigated using two synthetic tritiated photoaffinity analogues incorporating a benzophenone photoaffinity moiety at two different positions of the E7974 scaffold. Both analogues preferentially photolabeled alpha-tubulin, although minor binding to beta-tubulin was also detected. E7974 thus seems to share a unique, predominantly alpha-tubulin-targeted mechanism with other hemiasterlin-based compounds, suggesting that, unlike many tubulin-targeted natural products and related drugs, the hemiasterlins evolved to mainly target alpha-tubulin, not beta-tubulin subunits.
authors.affiliation: Division of Biology,, Eisai Research Institute, 4 Corporate Drive, Andover, MA 01810-2441, USA. galina_kuznetsov@eri.eisai.com
authors.forename: Galina
authors.initials: G
authors.lastname: Kuznetsov
authors.forename: Karen
authors.initials: K
authors.lastname: TenDyke
authors.forename: Murray J
authors.initials: MJ
authors.lastname: Towle
authors.forename: Hongsheng
authors.initials: H
authors.lastname: Cheng
authors.forename: Junke
authors.initials: J
authors.lastname: Liu
authors.forename: Joanne P
authors.initials: JP
authors.lastname: Marsh
authors.forename: Shawn E R
authors.initials: SE
authors.lastname: Schiller
authors.forename: Mark R
authors.initials: MR
authors.lastname: Spyvee
authors.forename: Hu
authors.initials: H
authors.lastname: Yang
authors.forename: Boris M
authors.initials: BM
authors.lastname: Seletsky
authors.forename: Christina J
authors.initials: CJ
authors.lastname: Shaffer
authors.forename: Veronique
authors.initials: V
authors.lastname: Marceau
authors.forename: Ye
authors.initials: Y
authors.lastname: Yao
authors.forename: Edward M
authors.initials: EM
authors.lastname: Suh
authors.forename: Silvio
authors.initials: S
authors.lastname: Campagna
authors.forename: Francis G
authors.initials: FG
authors.lastname: Fang
authors.forename: James J
authors.initials: JJ
authors.lastname: Kowalczyk
authors.forename: Bruce A
authors.initials: BA
authors.lastname: Littlefield
mesh_terms: D000818:Animals; D050256:Antimitotic Agents / Q000737:chemistry / Q000494:pharmacology*; D000970:Antineoplastic Agents / Q000737:chemistry / Q000494:pharmacology*; D017209:Apoptosis / Q000187:drug effects; D001688:Biological Products / Q000737:chemistry* / Q000494:pharmacology; D002417:Cattle; D045744:Cell Line, Tumor; D016195:G2 Phase / Q000187:drug effects; D006801:Humans; D008870:Microtubules / Q000187:drug effects / Q000378:metabolism; D008938:Mitosis / Q000187:drug effects; D009842:Oligopeptides / Q000737:chemistry* / Q000494:pharmacology*; D019920:Photoaffinity Labels; D010880:Piperidines / Q000737:chemistry / Q000494:pharmacology*; D011161:Porifera / Q000737:chemistry*; D012623:Seawater; D008941:Spindle Apparatus / Q000187:drug effects / Q000378:metabolism; D014404:Tubulin / Q000378:metabolism*; D014747:Vinblastine / Q000494:pharmacology
chemical_list: D050256:Antimitotic Agents; D000970:Antineoplastic Agents; D001688:Biological Products; C546192:E 7974; D009842:Oligopeptides; D019920:Photoaffinity Labels; D010880:Piperidines; D014404:Tubulin; D014747:Vinblastine; C467070:hemiasterlin