Abstract 45: <i>In vitro</i> and <i>in vivo</i> activity of a novel c-Met-targeting antibody-drug conjugate using a DNA-alkylating, indolinobenzodiazepine payload
Authors: Katharine C. Lai, Asli Muvaffak, Min Li, Marian Themeles, Surina Sikka, Kerry Donahue, Stuart W. Hicks, Angela Romanelli, Thomas Chittenden
Journal: Cancer Research (2017)

Abstract Purpose: c-Met dysregulation and/or overexpression are associated with tumor progression, metastasis and poor prognosis in numerous cancers. Despite strong pre-clinical evidence that blocking c-Met activity inhibits tumor cell growth and metastasis, targeted therapies have thus far failed to deliver an effective treatment option to the majority of patients. To address patients with both c-Met over-expressing and MET amplified tumors, we designed an antibody-drug conjugate (ADC) comprised of a humanized anti-c-Met monoclonal antibody linked to a highly potent indolinobenzodiazepine DNA-alkylating payload (DGN549) to enable activity against not only MET amplified but also c-Met over-expressing tumors. Experimental Design: Panels of monoclonal antibodies (Abs) against c-Met were generated and screened for antagonistic and agonistic activity in the presence or absence of the c-Met ligand, HGF. Lead Abs were humanized and conjugated to DGN549 either through lysine (Drug-to-Ab ratio (DAR) = 2.5) or engineered cysteine residues (DAR 2.0). Abs were also conjugated via lysine residues to the potent anti-microtubule maytansine derivative, DM4, using a sulfo-SPDB linker (DAR 3.5). Binding and cytotoxicity of ADCs were tested in vitro on normal and cancer cell lines with varying c-Met levels. Expression of c-Met was evaluated in patient tumors and xenografts along with normal human tissues using the CONFIRM immunohistochemistry assay. In vivo efficacy of anti-c-Met-DGN549 and anti-c-Met-DM4 ADCs was tested in both MET amplified and c-Met over-expressed (but non-amplified) xenograft tumor models. Results: A humanized anti-c-Met antibody, hucMet27, was identified which exhibits low c-Met agonist activity. Conjugates of hucMet27 were prepared with two different payloads, DGN549 and DM4, and in vitro and in vivo activity were determined. Both DGN549 and DM4 conjugates of hucMet27 bound with similar sub-nanomolar affinity to c-Met-expressing cells. hucMet27-DGN549 conjugates exhibited potent cytotoxicity against a large panel of c-Met expressing cell lines. By contrast, the potency of the hucMet27-DM4 conjugate was restricted mainly to cell lines harboring MET amplification, despite all cell lines demonstrating sensitivity to the unconjugated payload. When tested in mice bearing human xenograft tumors, both hucMet27-DGN549 and hucMet-DM4 conjugates were highly active in a MET amplified model, whereas hucMet27-DGN549 was more potent in inducing regressions in a model with c-Met over-expression without MET amplification. Conclusion: hucMet27-DGN549 exhibits compelling c-Met targeted anti-cancer activity in vitro and in vivo, and represents a promising therapeutic strategy to deliver a potent cytotoxic agent to tumor cells bearing a wide range of c-Met expression. Citation Format: Katharine C. Lai, Asli Muvaffak, Min Li, Marian Themeles, Surina Sikka, Kerry Donahue, Stuart W. Hicks, Angela Romanelli, Thomas Chittenden. In vitro and in vivo activity of a novel c-Met-targeting antibody-drug conjugate using a DNA-alkylating, indolinobenzodiazepine payload [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 45. doi:10.1158/1538-7445.AM2017-45