Discovery and characterization of two anti-PD-1 antibodies with a unique binding mechanism to human PD-1

Targeting checkpoint inhibitors is an effective therapy for treating cancer, with human programmed cell death protein 1 (hPD-1) being one of the most successful targets for developing antibody-based drugs. In this work, we isolated a panel of anti-PD-1 single-chain variable fragments with different binding and functional profiles from a fully synthetic human phage display library. Conversion of the best clone to hIgG1LALA and hIgG4PE formats, called UDIZ-007 and UDIZ-008, respectively, resulted in antibodies that effectively blocked the PD-1:PD-L1/L2 interaction and were highly selective as they did not cross-react with CD28 receptor family members. Doses of UDIZ-007 or UDIZ-008 at 10 mg/kg every 3 days for a total of six intraperitoneal administrations eradicated MC38-hPD-L1 colon tumors in B-hPD-1 transgenic mice for hPD-1 at day 17, with no relapse until the end of the study at day 56. Importantly, these antibodies bind hPD-1 in a unique region compared to the anti-PD-1 antibodies of known structure, which might have an impact on novel oncology indications when used as a standalone therapy or in combination with currently approved anti-PD-1 therapeutic antibodies. Therefore, UDIZ-007 and UDIZ-008 seem to be promising candidates for the development of antibody-based drugs targeting checkpoint inhibitors as a treatment for cancer.