Abstract 5100: Combination of κλ bispecific antibodies targeting innate (CEAxCD47, NILK-2401) and adaptive immunity (CEAxCD3, NILK-2301 and CEAxCD28, NILK-3301) for next generation immunotherapy of CEA-expressing cancers

• Published in: Cancer research (Chicago, Ill.) Ill.), 2023-04, Vol.83 (7_Supplement), p.5100-5100
• Main Authors: Seckinger, Anja, Nouveau, Lise, Majocchi, Sara, Moine, Valéry, Buatois, Vanessa, Daubeuf, Bruno, Gueneau, Franck, Ravn, Ulla, Masternak, Krzysztof, Poitevin, Yves, Rousset, Emeline, Magistrelli, Giovanni, Malinge, Pauline, Shang, Limin, Fischer, Nicolas, Strein, Klaus, Ferlin, Walter, Hose, Dirk
• Format: Article
• Language: English
• ISSN: 1538-7445; 1538-7445
• DOI: 10.1158/1538-7445.AM2023-5100

Background: The CEAxCD3 bispecific antibody (bsAb) NILK-2301 couples CEA (CEACAM5) on cancer cells and CD3 on T-cells inducing T-cell activation (signal 1) and tumor cell killing (TDCC). T-cell activation can be boosted by CEA-targeted CD28-costimulation (NILK-3301; signal 2). NILK-2401, carrying a fully effective IgG1 Fc, induces antibody-dependent phagocytosis (ADCP) and antibody-dependent cytotoxicity (ADCC) of tumor cells by co-targeting CEA and the innate immune checkpoint CD47 (“don’t eat me” signal). We present here next generation immunotherapy to overcome limited single class activity in CEA-expressing solid cancers. Methods: BsAbs were generated using LCB’s fully human κλ body platform. TDCC, ADCP, and ADCC with human PBMC or monocyte-derived macrophages were assessed using CEA+ colorectal (n=3), lung (n=2), and gastric (n=2) cancer lines. Combination activity of NILK-2401 + NILK-2301 (± NILK-3301) was assessed by flow cytometry. In vivo activity was tested in xenograft NOG or NSG/human PMBC-, HIS-, and hSIRPα/hCD47/hCD3/hCD28 transgenic mice. Safety data include binding to other CEACAMs, cytokine release in whole blood, erythrophagocytosis, platelet activation, exclusion of superagonism (NILK-3301), as well as PK- and tolerability in cynomolgus monkeys and Tg32-mice. Results: NILK-2301 induced dose-dependent killing of all tested cell lines, which was also visualized by live cell imaging. Combination of NILK-2301 (1 nM) + NILK-3301 vs. NILK-2301 alone (10 nM) increased TDCC (3-8-fold), T-cell activation (CD25, CD69, HLA-DR), cytokine secretion (interferon-γ, granzyme B, perforin), and CD4+/CD8+ T-cell proliferation. NILK-2401 blocked CD47-SIRPα interaction and induced ADCP/ADCC-mediated elimination of all cell lines. NILK-2301 + NILK-2401 treatment increased maximum activity (Emax) and reduced necessary dose of the T-cell bsAb to reach Emax. E.g., Emax of 30% killing (NILK-2301 alone) was increased in combination with NILK-2401 at 0.1/1/10 µg/mL to 40%, 80%, and 80%. In vivo, NILK-2301 (10 mg/kg IV, BIW) decreased tumor progression. NILK-2301/-3301 combination induced tumor regression in 8/8 mice. NILK-2401 delayed tumor growth vs. mean of control in 100% (15/15) of mice and prevented establishment of detectable tumors (>50mm3) in 53% (8/15). Results of double and quadruple transgenic mice, including triple bsAb combinations, will be presented at the meeting. No relevant safety signals were detected. Conclusions: NILK-2301 and NILK-2401 are acti