Role of thymidylate synthase in the antitumor activity of the multitargeted antifolate, LY231514.
The cytotoxicity of LY231514 was only partially alleviated by thymidine addition (5 microM) in GC3 human colon carcinoma cells, and complete protection required the addition of both hypoxanthine (100 microM) and thymidine. In contrast, the cytotoxic activity of tomudex (raltitrexed, ZD1694) was completely reversed by thymidine alone. MCF-7 human breast and H630 human colon carcinoma cells selected for resistance to tomudex and 5-fluorouracil, respectively via thymidylate synthase (TS) amplification demonstrated only modest resistance to LY231514 compared to tomudex. LY231514-induced cytotoxicity in these resistant cell lines was completely prevented by the addition of hypoxanthine (100 microM), indicating inhibition of purine de novo biosynthesis as a secondary target for LY231514 action. Thymidine at physiologic levels in mouse plasma (approximately 1 microM) produced only a 2.6-fold shift in the IC50 for LY231514-mediated cytotoxicity in GC3/cl1 cells compared to a 128-fold shift for tomudex. LY231514 treatment (i.p., qd x 10) significantly delayed tumor growth in the GC3 carcinoma xenograft model. However, a thymidine kinase-deficient mutant of this same tumor line demonstrated heightened sensitivity to the in vivo antitumor activity of LY231514 with complete regression of established tumors and a large number of tumor-free survivors after one course of treatment. The data demonstrate that inhibition of thymidylate synthase is a prominent mechanism for antitumor activity by LY231514, but important secondary sites of action exist for this multitargeted molecule.
Schultz
R M
RM
Lilly Research Laboratories, Eli Lilly and Company, Indianapolis, Indiana 46285, USA. Schultz_Richard_M@Lilly.Com
Patel
V F
VF
Worzalla
J F
JF
Shih
C
C
D000818:Animals; D000970:Antineoplastic Agents / Q000494:pharmacology*; D003110:Colonic Neoplasms / Q000188:drug therapy; D019008:Drug Resistance, Neoplasm; D004791:Enzyme Inhibitors / Q000494:pharmacology*; D005493:Folic Acid Antagonists / Q000494:pharmacology*; D005971:Glutamates / Q000494:pharmacology*; D006147:Guanine / Q000031:analogs & derivatives* / Q000494:pharmacology; D006801:Humans; D051379:Mice; D000068437:Pemetrexed; D011799:Quinazolines / Q000494:pharmacology; D013876:Thiophenes / Q000494:pharmacology; D013936:Thymidine / Q000494:pharmacology; D013940:Thymidylate Synthase / Q000502:physiology*; D014407:Tumor Cells, Cultured
D000970:Antineoplastic Agents; D004791:Enzyme Inhibitors; D005493:Folic Acid Antagonists; D005971:Glutamates; D011799:Quinazolines; D013876:Thiophenes; D000068437:Pemetrexed; D006147:Guanine; D013940:Thymidylate Synthase; C068874:raltitrexed; D013936:Thymidine