Application of anti-methotrexate Fab fragments for the optimization of intraperitoneal methotrexate therapy in a murine model of peritoneal cancer

Anti-drug antibodies may be used to impart regio-specific alterations in drug disposition, potentially enhancing the therapeutic selectivity of intracavitary chemotherapy. In the present study, we tested the hypotheses that systemic therapy with anti-methotrexate antibodies would allow increases in the maximum tolerated dose of intraperitoneal methotrexate (MTX) and allow increases in the therapeutic efficacy of intraperitoneal MTX in a murine model of peritoneal cancer. Monoclonal anti-MTX Fab antibody fragments (AMF) were produced, purified, and characterized. AMF pharmacokinetics were determined following i.v. bolus injection (0.4 g/kg) and s.c. bolus injection (0.4, 0.8, 2.2 g/kg). MTX efficacy was investigated in mice bearing peritoneal sarcoma 180 tumors, following administration of MTX via 72 h i.p. infusion at 1.9, 2.8, 3.8 mg/kg, and following combination therapy of 7.5 or 10 mg/kg i.p. MTX (72 h infusion) and 4.2 g/kg s.c. AMF. The mean terminal half-life of AMF was found to be 10.9 ± 3.3 h and was not dose-dependent, and s.c. bioavailability was 28% ± 7% at 2.2 g/kg. In mice bearing peritoneal tumors, the maximally tolerated dose of i.p. MTX increased from 1.9 mg/kg (following i.p. MTX alone) to 10 mg/kg (with co-administration of s.c. AMF). Median survival times for saline-treated control animals and animals receiving i.p. MTX(1.9, 2.8, 3.8 mg/kg) were 9, 12, 10, and 7 days, respectively. However, for animals receiving combination therapy with i.p. MTX 7.5 or 10 mg/kg and 4.2 g/kg s.c. AMF, median survival time increased to 17 and 14 days, respectively. As such, the present data suggest that systemic administration of AMF may allow increases in the maximally tolerated dose of i.p. MTX, and allow increases in the therapeutic efficacy of i.p. MTX chemotherapy of peritoneal tumors.