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1210 A human bispecific antibody targeting LAG-3 and PD-1 (INCA32459) potently activates exhausted T cells
  1. Shaun Stewart1,
  2. Floris Fransen2,
  3. Shane Harvey1,
  4. Franziska Mortensen2,
  5. Anita Stam2,
  6. Rahel Awdew1,
  7. Arpita Mondal1,
  8. Christina Stevens1,
  9. Eric Rovers2,
  10. Steef Engels2,
  11. Melissa Rentrop-Boeijen2,
  12. Linda Hendriks2,
  13. Brenda van Dieren2,
  14. Rebecca Buonpane1,
  15. Therese Visser2,
  16. Pepijn Schellen2,
  17. Ashwini Kulkarni1,
  18. Jonathan Rios-Doria1,
  19. Jing Zhou1,
  20. Paul Tacken2,
  21. Lu Lu1,
  22. Vanessa Zondag-van der Zande2,
  23. Cheng-Yen Huang1,
  24. Renate den Blanken-Smit2,
  25. John de Kruif2,
  26. Rinse Klooster2,
  27. Simon Plyte2,
  28. Horacio Nastri1 and
  29. Patrick Mayes1
  1. 1Incyte Research Institute, Wilmington, DE, USA
  2. 2Merus N.V, Utrecht, Netherlands


Background Exhausted T cells are characterized by the expression of negative immune regulatory receptors, including programmed death protein-1 (PD-1) and lymphocyte-activation gene 3 (LAG-3), which inhibit the proliferation and function of T cells and limit antitumor immunity. We describe the generation and characterization of INCA32459, a human IgG1 Fc-silenced bispecific antibody that simultaneously binds to PD-1 and LAG-3 and reverts their inhibitory function.

Methods INCA32459 was generated using the Merus common light chain Biclonics® platform. LAG-3 and PD-1 Fab panels were generated through immunization of Merus MeMo® mice, and large panels of Biclonics® libraries were screened before optimizing lead candidate molecules.

Results INCA32459 binds with high affinity to both human (KD=0.39 nM) and cynomolgus monkey (KD=0.44 nM) PD-1, and human (KD=1.15 nM) and cynomolgus monkey (KD=0.20 nM) LAG-3, as measured by surface plasmon resonance. The monovalent PD-1 arm of INCA32459 blocks PD-1 with similar potency as a bivalent PD-1 antibody (nivolumab analog) in a PD-1/PD-L1 reporter assay. In a loss-of-function reporter assay where luciferase expression increases upon blockade of both LAG-3 and PD-1, INCA32459 significantly induced luciferase expression to a greater extent than either PD-1 (nivolumab analog) or LAG-3 (relatlimab analog) single agent antibody controls, and greater than PD-1 (nivolumab) and LAG-3 (relatlimab) analog antibodies combined. In 2 human primary immune cell assays, a T-cell exhaustion model using chronically SEB-stimulated peripheral blood mononuclear cells (PBMCs), and an antigen recall assay using CEFT MHCII peptide pool-stimulated PBMCs, INCA32459 treatment resulted in higher interleukin-2 and interferon-y induction, respectively, compared with PD-1 (nivolumab analog) and LAG-3 (relatlimab analog) antibodies combined. In a human MDA-MB-231 breast tumor model in CD34+ humanized NSG mice, INCA32459 treatment decreased tumor growth compared with a combination of PD-1 (pembrolizumab) and LAG-3 (relatlimab analog) antibodies. Pharmacodynamic analysis in mice demonstrated a dose-dependent increase in receptor occupancy at 1 and 10 mg/kg. Pharmacokinetic characterization of INCA32459 in cynomolgus monkeys after a single IV infusion at 3 and 30 mg/kg demonstrated an average clearance, steady-state volume of distribution, and mean residence time of 0.515 mL/h/kg, 74.1 mL/kg, and 144 h, respectively.

Conclusions We have developed INCA32459, a potent dual inhibitor of PD-1 and LAG-3 in preclinical models, which induces activation of exhausted T cells to a greater extent than a combination of bivalent monospecific antibodies targeting PD-1 (nivolumab analog) and LAG-3 (relatlimab analog). These data support the clinical evaluation of INCA32459, and a phase 1 study in cancer patients is underway.

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