Inverse targeting of peritoneal tumors: Selective alteration of the disposition of methotrexate through the use of anti-methotrexate antibodies and antibody fragments

We have hypothesized that antidrug antibodies (ADAb) may be employed to impart site-specific alterations in the disposition of drug molecules, potentially allowing for targeted drug therapy. We are specifically interested in minimizing systemic exposure to free drug and systemic toxicities resultant from regional chemotherapy through the intravenous administration of ADAb. In this report, we present the production and purification of anti-methotrexate Fab fragments, and we present investigations of the effects of anti-methotrexate Fab and anti-methotrexate immunoglobulin G on the disposition of methotrexate in the rat. Pharmacokinetic studies revealed that intravenous anti-methotrexate immunoglobulin G (anti-MTX IgG) and anti-methotrexate Fab (anti-MTX Fab) administration produced dramatic alterations in the plasma pharmacokinetics of methotrexate (MTX), following intraperitoneal MTX administration (area under the total MTX concentration vs time curve for anti-MTX IgG relative to control, 420 ± 90 (p < 0.05); for anti-MTX Fab relative to control, 46 ± 6.1 (p < 0.05); area under the free MTX concentration vs time curve for anti- MTX IgG relative to control, 0.64 ± 0.16; for anti-MTX Fab relative to control, 0.45 ± 0.20 (p < 0.05)). Additional studies conducted in anesthetized rats revealed no significant alterations in the area under the total peritoneal MTX concentration vs time curves, free MTX peritoneal concentration vs time curves, or peritoneal exit rate of MTX in anti-MTX Fab treated animals relative to controls. Therefore, our pharmacokinetic studies demonstrate that ADAb may produce site-specific alterations in drug pharmacokinetics, potentially enhancing the site specificity of drug distribution and drug action following regional chemotherapy.