The role of cellular folates in the enhancement of activity of the thymidylate synthase inhibitor 10-propargyl-5,8-dideazafolate against hepatoma cells in vitro by inhibitors of dihydrofolate reductase.
Exposure of growing cultures of hepatoma cells in vitro to the lipid-soluble dihydrofolate reductase inhibitors metoprine (36 nM) or trimetrexate (2 nM) at subtoxic concentrations causes little change in cell growth rate, colony forming ability, cell cycle distribution, and de novo purine and thymidylate biosynthesis. The reductase inhibitors augment the cytotoxic activity of the thymidylate synthase inhibitor, 10-propargyl-5,8-dideazafolate by nearly 10-fold under optimal conditions. Treatment of the hepatoma cells with the reductase inhibitors for 72 h during growth caused approximately a 75% reduction in total cellular folates and 5,10-methylenetetrahydrofolate (primarily as polyglutamates) the substrate for thymidylate synthase. The reductase inhibitors also cause a doubling in the accumulation of 10-propargyl-5,8-dideazafolate polyglutamates. The combined antifolate treatment (metoprine or trimetrexate plus 10-propargyl-5,8-dideazafolate) expands the dUMP pool by 30-fold, which is more than the sum of either of the antifolates alone. Consequently, it is postulated that the enhanced activity of 10-propargyl-5,8-dideazafolate in combination with low concentrations of dihydrofolate reductase inhibitors is due to an increase in the ratio of inhibitor to substrate for thymidylate synthase of nearly 10-fold and an extensive enhancement of the dUMP pool. These conditions predispose the target enzyme and the cells to more effective metabolic blockade by 10-propargyl-5,8-dideazafolate which is presumably caused by the formation of an inhibited 10-propargyl-5,8-dideazafolate[polyglutamate]-thymidylate synthase-dUMP ternary complex.
Galivan
J
J
Wadsworth Center for Laboratories and Research, New York State Department of Health, Albany 12201.
Rhee
M S
MS
Johnson
T B
TB
Dilwith
R
R
Nair
M G
MG
Bunni
M
M
Priest
D G
DG
D000818:Animals; D000970:Antineoplastic Agents / Q000494:pharmacology*; D002455:Cell Division / Q000187:drug effects; D002460:Cell Line; D005492:Folic Acid / Q000031:analogs & derivatives* / Q000378:metabolism* / Q000494:pharmacology; D005493:Folic Acid Antagonists / Q000494:pharmacology*; D007700:Kinetics; D008114:Liver Neoplasms, Experimental / Q000473:pathology*; D011739:Pyrimethamine / Q000031:analogs & derivatives / Q000494:pharmacology; D011799:Quinazolines / Q000494:pharmacology*; D051381:Rats; D013940:Thymidylate Synthase / Q000037:antagonists & inhibitors*; D016597:Trimetrexate
D000970:Antineoplastic Agents; D005493:Folic Acid Antagonists; D011799:Quinazolines; C006308:metoprine; C031662:CB 3717; D005492:Folic Acid; D013940:Thymidylate Synthase; D016597:Trimetrexate; D011739:Pyrimethamine