Reduced forms of aminopterin and methotrexate in L1210 lymphoma

We examined several characteristics of reduced forms of folate antagonists. Dihydro and tetrahydro derivatives of aminopterin and methotrexate (MTX) were chemically prepared. When titrated with dihydrofolate reductase, reduced derivatives were equipotent with the parent compounds and titrated stoichiometrically with the enzyme at pH 6 and nonstoichiometrically at higher pH conditions (pH 7.4 and above). When incubated with L1210 cells in vitro, rates of uptake of tritiated reduced compounds were significantly less in L1210 and L1210/MTX cells compared with the respective oxidized parent compounds and significantly less in L1210/MTX than in L1210 cells. Although dihydroaminopterin and tetrahydromethotrexate increased the survival rate of mice bearing L1210 tumors, these compounds had no such effect on the life-spans of animals with L1210/MTX tumors.