Tubulin-dependent hydrolysis of guanosine triphosphate as a screening test to identify new antitubulin compounds with potential as antimitotic agents: application to carbamates of aromatic amines.
Tubulin-dependent GTP hydrolysis was evaluated for its potential as a relatively simple screening assay for new antimitotic drugs. Carbamates of aromatic amines were chosen as the test system because of the relatively diverse structures of compounds in this class already known to have antimitotic properties and because of the large number of such compounds in the NSC collection of the National Cancer Institute. Of 162 compounds evaluated, significant alterations in the GTPase reaction were observed with 26 agents. Sixteen of these had substantial inhibitory effects on tubulin polymerization (true positives), while ten did not (false positives). There were no false negatives (i.e., no agent inactive in the GTPase assay inhibited tubulin polymerization). The true positives were examined for effects on cell growth and mitosis, and four compounds had 50% inhibitory concentration values of 2 microM or less with L1210 murine leukemia cells. All four caused the accumulation of cells in metaphase arrest. We conclude that tubulin-dependent GTP hydrolysis can be used effectively to select new antitubulin compounds with potential as antimitotic agents from a large group of compounds of unknown activity.
Chi
D M
DM
Laboratory of Biochemical Pharmacology, National Cancer Institute, Bethesda, Maryland 20892.
Shahrik
L K
LK
Ho
H H
HH
Hamel
E
E
D000970:Antineoplastic Agents / Q000494:pharmacology*; D001562:Benzimidazoles / Q000494:pharmacology*; D020558:GTP Phosphohydrolases / Q000032:analysis; D006160:Guanosine Triphosphate / Q000378:metabolism*; D006868:Hydrolysis; D008938:Mitosis / Q000187:drug effects*; D015739:Nocodazole; D011719:Pyrazines / Q000494:pharmacology*; D011799:Quinazolines / Q000494:pharmacology*; D013329:Structure-Activity Relationship; D050257:Tubulin Modulators
D000970:Antineoplastic Agents; D001562:Benzimidazoles; D011719:Pyrazines; D011799:Quinazolines; D050257:Tubulin Modulators; C038202:NSC 251635; C032130:NSC 181928; C051598:NSC 330770; D006160:Guanosine Triphosphate; D020558:GTP Phosphohydrolases; D015739:Nocodazole