DuoBody-CD3xCD20 induces potent T-cell-mediated killing of malignant B cells in preclinical models and provides opportunities for subcutaneous dosing

Engelberts, P.J.; Hiemstra, I.H.; Jong, B. de; Schuurhuis, D.H.; Meesters, J.; Hernandez, I.B.; ... ; Breij, E.C.W. (2020)

Article / Letter to editor

Background: DuoBody (R)-CD3xCD20 (GEN3013) is a full-length human IgG1 bispecific antibody (bsAb) recognizing CD3 and CD20, generated by controlled Fab-arm exchange. Its Fc domain was silenced by introduction of mutations L234F L235E D265A.Methods: T-cell activation and T-cell-mediated cytotoxicity were measured by flow cytometry following co-culture with tumour cells. Anti-tumour activity of DuoBody-CD3xCD20 was assessed in humanized mouse models in vivo. Non-clinical safety studies were performed in cynomolgus monkeys.Findings: DuoBody-CD3xCD20 induced highly potent T-cell activation and T-cell-mediated cytotoxicity towards malignant B cells in vitro. Comparison of DuoBody-CD3xCD20 to CD3 bsAb targeting alternative B-cell antigens, or to CD3xCD20 bsAb generated using alternative CD20 Ab, emphasized its exceptional potency. In vitro comparison with other CD3xCD20 bsAb in clinical development showed that DuoBody-CD3xCD20 was significantly more potent than three other bsAb with single CD3 and CD20 binding regions and equally potent as a bsAb with a single CD3 and two CD20 binding regions. DuoBody-CD3xCD20 showed promising anti-tumour activity in vivo, also in the presence of excess levels of a CD20 Ab that competes for binding. In cynomolgus monkeys, DuoBody-CD3xCD20 demonstrated profound and long-lasting B-cell depletion from peripheral blood and lymphoid organs, which was comparable after subcutaneous and intravenous administration. Peak plasma levels of DuoBody-CD3xCD20 were lower and delayed after subcutaneous administration, which was associated with a reduction in plasma cytokine levels compared to intravenous administration, while bioavailability was comparable.Interpretation: Based on these preclinical studies, a clinical trial was initiated to assess the clinical safety of subcutaneous DuoBody-CD3xCD20 in patients with B-cell malignancies. (C) 2020 The Authors. Published by Elsevier B.V.

• Bispecific antibody
• CD3
• CD20
• T cell redirection
• B cell malignancy
• Subcutaneous administration

• All authors: Engelberts, P.J.; Hiemstra, I.H.; Jong, B. de; Schuurhuis, D.H.; Meesters, J.; Hernandez, I.B.; Oostindie, S.C.; Neijssen, J.; Brink, E.N. van den; Horbach, G.J.; Verploegen, S.; Labrijn, A.F.; Salcedo, T.; Schuurman, J.; Parren, P.W.H.I.; Breij, E.C.W.
• Date: 2020-02-01
• Journal: EBioMedicine
• Volume: 52
• DOI: doi:10.1016/j.ebiom.2019.102625
• Link: https://www.sciencedirect.com/science/article/pii/S2352396419308400?via%...