Inhibition of Dihydrofolate Reductase and Cell Growth by Antifolates in a Methotrexate-Resistant Cell Line

The structural features and lipid solubility of four different classes of antifolate compounds were compared for their inhibition of dihydrofolate reductase (DHFR) and growth in a normal and methotrexate (MTX)-resistant 3T6 mouse cell line. All of the compounds have been shown previously to have antifolate activity. The resistant cell line has a 7-fold increase in DHFR activity with normal transport, but an altered affinity for MTX. All the antifolates were equally effective in inhibiting DHFR and growth in the parent cell line. Inhibition of partially purified DHFR from the resistant cells increased with changes in lipid solubility and structure of the compounds, compared to the parent DHFR. These data demonstrate that the resistant cells may be more sensitive to the structurally dissimilar antifolates than to MTX and lend importance to further development of this type of antifolate. These results suggest that these compounds may be useful in circumventing antifolate resistance due to alterations in target enzyme concentration and drug-enzyme affinity, as well as drug transport.