The glutamine analog acivicin as antipyrimidine. Studies on the interrelationship between pyrimidine and urea synthesis in liver.
The inhibition of cytosolic carbamoyl-phosphate synthetase II by acivicin was used to study the role of the cytosolic carbamoyl phosphate pool as the exclusive substrate source for de novo pyrimidine synthesis in rat hepatocytes. De novo pyrimidine synthesis was stimulated: 1. by uridine triphosphate deficiency (incubation with D-galactosamine) leading to a stimulation of cytosolic carbamoyl phosphate synthesis, and 2. by accumulation and efflux of mitochondrial carbamoyl phosphate (incubation with ammonium ions and L-norvaline). The stimulated orotate formation from cytosolic carbamoyl phosphate in UTP depleted cells was completely blocked by acivicin. It was not influenced by an inhibition of mitochondrial carbamoyl phosphate synthesis mediated by 4-pentenoate, since mitochondrial carbamoyl phosphate did not participate in cytosolic pyrimidine synthesis even in the presence of ammonium ion concentrations maintaining physiological rates of urea synthesis. An excess of ammonium ions led to an artificial accumulation and efflux of mitochondrial carbamoyl phosphate, which could be avoided by 4-pentenoate. The non-regulated stimulation of pyrimidine synthesis from surplus mitochondrial carbamoyl phosphate was not inhibited by acivicin. Utilization of mitochondrial carbamoyl phosphate for de novo pyrimidine synthesis presumably does not occur under physiological conditions because mitochondrial CP efflux depends on the accumulation of this metabolite in the mitochondria under experimental or pathological circumstances. Acivicin inhibition of CPS II thus cannot be bypassed by mitochondrial CP. It is suitable as inhibitor of the physiological de novo pyrimidine synthesis.
D000818:Animals; D000903:Antibiotics, Antineoplastic / Q000494:pharmacology*; D002221:Carbamyl Phosphate / Q000378:metabolism; D003600:Cytosol / Q000378:metabolism; D005260:Female; D007555:Isoxazoles / Q000494:pharmacology*; D008099:Liver / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D008930:Mitochondria, Liver / Q000378:metabolism; D009963:Orotic Acid / Q000378:metabolism; D010080:Oxazoles / Q000494:pharmacology*; D011743:Pyrimidines / Q000037:antagonists & inhibitors*; D051381:Rats; D011919:Rats, Inbred Strains; D014508:Urea / Q000096:biosynthesis*
D000903:Antibiotics, Antineoplastic; D007555:Isoxazoles; D010080:Oxazoles; D011743:Pyrimidines; D002221:Carbamyl Phosphate; D009963:Orotic Acid; D014508:Urea; C030986:pyrimidine; C006303:acivicin