Anti pd 1 mab

Panc-02 cells (1 × 10⁶ cells) were subcutaneously implanted in C57BL/6 mice on day 0, and starting on day 16 when tumor diameters were approximately 4 mm in diameter, mice were treated with anti-PD-1 mAb (BioXcell, Lebanon, NH) (200 μg, intraperitoneal (ip) injection) and/or DHA-dFdC (100 mg/kg, ip). anti-PD-1 mAb therapy was administered on days 16, 23, 27, 30, 34, and 37 and DHA-dFdC was administered on days 16, 23, 30, and 37 to their respective groups. Mouse health was monitored, and tumor size was measured using a digital caliper. Mice were euthanized four days after the last treatment.

For the in vivo studies, 6-week-old, female Balb/C mice from The Jackson Laboratory were used. A total of 1 × 10⁵ 4T1 wild-type cells were injected in the mammary fat pad of each mice. The tumors were palped every 3 days, and body weight and tumor volumes were measured. At the end of the 6 weeks or when tumor size reached 2 cm, the mice were sacrificed, and tumors and lungs were harvested. Debio-025 was administered at indicated concentrations every 3 days via oral gavage. No significant differences in the mice body weight were observed because of drug treatment. Anti-PD-1 mAb (BioXCell, clone: 29F.1A12) or isotype IgG antibodies were administered (intraperitoneally) every 3 days at 200 mg/kg/day dosage in the combination experiments with a total of four administrations per group/study starting at day 10. For estimation of metastasis, lungs were washed with PBS thrice and added to Bouin solution. Pulmonary metastases were counted using stereomicroscope by two investigators separately. All the procedures involving animal care use were approved by IACUC of Rutgers University (Newark, NJ).