Cell-based screen for antimitotic agents and identification of analogues of rhizoxin, eleutherobin, and paclitaxel in natural extracts.
We describe a cell-based assay for antimitotic compounds that is suitable for drug discovery and for quantitative determination of antimitotic activity. In the assay, cells arrested in mitosis as a result of exposure to antimitotic agents in pure form or in crude natural extracts are detected by ELISA using the monoclonal antibody TG-3. The assay was used to screen >24,000 extracts of marine microorganisms and invertebrates and terrestrial plants and to guide the purification of active compounds from 5 of 119 positive extracts. A new rhizoxin analogue was found in a Pseudomonas species, six new eleutherobin analogues were identified from the octocoral Erythropodium caribaeorum, and two paclitaxel analogues were found in the stem bark of the tree Ilex macrophylla. The assay was also used for quantitative comparison of the antimitotic activity of different analogues. It revealed the importance of the C-11 to C-13 segment of the diterpene core of eleutherobin for its antimitotic activity. The identification of antimitotic compounds in very low abundance and their high (0.5%) occurrence in natural extracts indicates that drug discovery efforts using this cell-based assay may lead to the identification of structurally novel antimitotic agents.
Roberge
M
M
Department of Biochemistry and Molecular Biology, University of British Columbia, Vancouver, Canada. michel@otter.biochem.ubc.ca
Cinel
B
B
Anderson
H J
HJ
Lim
L
L
Jiang
X
X
Xu
L
L
Bigg
C M
CM
Kelly
M T
MT
Andersen
R J
RJ
D000470:Alkaloids / Q000302:isolation & purification / Q000494:pharmacology*; D000818:Animals; D000903:Antibiotics, Antineoplastic / Q000302:isolation & purification / Q000494:pharmacology; D000970:Antineoplastic Agents / Q000302:isolation & purification / Q000494:pharmacology*; D000972:Antineoplastic Agents, Phytogenic / Q000302:isolation & purification / Q000494:pharmacology; D001943:Breast Neoplasms / Q000188:drug therapy / Q000473:pathology; D004224:Diterpenes; D004354:Drug Screening Assays, Antitumor / Q000379:methods*; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D007448:Invertebrates / Q000737:chemistry; D007783:Lactones / Q000302:isolation & purification / Q000494:pharmacology*; D018942:Macrolides; D008386:Marine Biology; D017239:Paclitaxel / Q000031:analogs & derivatives* / Q000302:isolation & purification; D010936:Plant Extracts / Q000302:isolation & purification / Q000494:pharmacology; D010946:Plants, Medicinal / Q000737:chemistry; D011549:Pseudomonas / Q000737:chemistry; D013329:Structure-Activity Relationship; D014020:Tissue Extracts / Q000302:isolation & purification / Q000494:pharmacology; D014407:Tumor Cells, Cultured / Q000187:drug effects
D000470:Alkaloids; D000903:Antibiotics, Antineoplastic; D000970:Antineoplastic Agents; D000972:Antineoplastic Agents, Phytogenic; D004224:Diterpenes; D007783:Lactones; D018942:Macrolides; D010936:Plant Extracts; D014020:Tissue Extracts; C109551:eleutherobin; C041443:rhizoxin; D017239:Paclitaxel