The antitumor activity of dihydrohomofolate against L1210 and L1210/FR-8 leukemia was studied and found to be comparable to that of tetrahydrohomofolate. Pretreatment of mice with methotrexate. a potent inhibitor of dihydrofolate reductase, decreased the antitumor activity of reduced homofolates against L1210/FR-8 indicating that an adequate level of free enzyme is probably necessary for the antitumor activity. The reduced homofolates were found to be less effective against L1210 than against L1210/FR-8, however, the antitumor activity became comparable when mice were kept on a folate-deficient diet. This suggests that factors other than tissue levels of dihydrofolate reductase could influence the response of tumors to reduced homofolates.

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