Abstract 4479: BT8009: A bicyclic peptide toxin conjugate targeting Nectin-4 (PVRL4) displays efficacy in preclinical tumor models

Abstract

Abstract We have identified a Nectin-4 targeting peptide for delivery of cytotoxic agents to Nectin-4 expressing tumors. Nectin-4 is a cell adhesion molecule, that is highly expressed in certain tumor types, including bladder, TNBC and NSCLC, but has a restricted distribution in normal tissue. Bicycles® are small (1.5kDa) fully synthetic, structurally constrained, peptide drugs that combine the affinity of antibodies with the pharmacokinetic properties of small molecules. The Bicycle phage display platform was utilised to rapidly identify a high affinity (18 nM) and highly selective bicyclic Nectin-4 binding peptide. Synthetic modification of the initial lead peptide improved affinity (0.3 nM), hydrophilicity and stability. The optimised peptide is conjugated through an inert sarcosine spacer chain and a cleavable linker to the toxin MMAE, to form the Bicycle Toxin Conjugate (BTC) BT8009. Once bound to cell surface Nectin-4, the linker system is cleaved by peptidases (e.g. cathepsin B) upregulated in the tumor micro environment. Full structure of BT8009 will be disclosed within the presentation. Fluorescence polarisation and surface plasmon resonance show BT8009 has low nanomolar affinity (3nM) for Nectin-4 and high selectivity (>1000 fold) over Nectins 1-3, and the 5 nectin-like, family members. Good affinity for the cognate native protein is maintained across the preclinical safety species. High content imaging of cultures of MDA-MB-468 cells, demonstrates binding of the BTC to cell membrane. In vivo, BT8009 is well tolerated in mouse and rat and shows regressions across a range of cell and patient derived xenograft models. Efficacy correlates with the expression level of Nectin-4 on the tumor cells, and the dose delivered. In the MDA-MB-468 (TNBC) CDX model full tumor regression was seen with weekly i.v. administration of 3 mg/k g i.v. (4 doses), with no tumor regrowth out to 70 days post last dose. Similar efficacy was seen in two NSCLC PDX models (adenocarcinoma and squamous cell carcinoma) with full regression attained in both. Near full regression was seen in a squamous cell esophageal cancer PDX. Larger tumors show a similar degree of sensitivity to the BTC, with rapid tumor regression from a volume of 800 mm3. As a small peptide BT8009 undergoes rapid clearance from the plasma with minimal exposure to non-targeted organs. After a single dose, MMAE has been shown to be retained within tumor tissue in excess of 60 h, at exposure levels significantly greater than corresponding plasma and other tissue levels. Bicycle Toxin Conjugates represent a novel treatment modality for nectin-4 expressing tumors with excellent efficacy in several mouse xenograft models. Citation Format: Mike Rigby, Paul Beswick, Gemma Mudd, Katerine Van Rietschoten, Liuhong Chen, Sophie M. Watcham, Heather Allen, Amy Brown, Helen Harrison, Gavin Bennett, Phil Jeffrey, Peter U. Park, Maria Koehler, Nicholas Keen. BT8009: A bicyclic peptide toxin conjugate targeting Nectin-4 (PVRL4) displays efficacy in preclinical tumor models [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 4479.