Abstract LB-232: APG350, a dimerized single chain TRAIL receptor agonist with enhanced functional properties
Authors: Gieffers Christian , Kluge Michael , Hill Oliver , Meinolf Thiemann , Merz Christian , Branschädel Marcus , Harald Fricke 
Published: June 2012

Abstract
TRAIL (Apo2L) is a member of the Tumor Necrosis Factor Superfamily (TNF-SF) that induces apoptosis through binding to two closely related receptors, TRAIL-R1 and TRAIL-R2. Its unique ability of selective apoptosis induction in tumor cells makes TRAIL an attractive molecule for tumor therapy.
APG350 is a newly designed TRAIL-receptor agonist comprising two single-chain TRAIL-receptor binding domains (each consisting of three covalently linked TRAIL protomers) that are dimerized via the Fc-part of human IgG1. APG350 has six receptor binding sites per molecule and binds to both death inducing TRAIL receptors. In vitro, APG350 shows potent apoptosis induction on a wide subset of human tumor cell-lines, on cancer stem cells and on primary tumor cells. Mechanistically its improved ability to form clusters on target cells composed of six TRAIL-receptors each, distinguishes APG350 from current clinical development candidates. Agonistic antibodies like Conatumumab or Apomab are capable to bind two receptor chains per molecule, while recombinant TRAIL (e.g. Dulanermin) binds three receptor chains per molecule. Direct in vitro comparison of APG350 with recombinant TRAIL (APG400) and a TRAIL-R2 specific agonistic monoclonal antibody revealed superior apoptosis induction capacity for APG350.
Analysis of the PK-parameters in mice showed a half life of 23.1h for APG350 (1.04h for APG400) indicating a significantly improved PK in comparison to APG400. The half life of APG350 in a Cynomolgus monkey was 26.7h.
Comparative treatment of mice bearing Colo205 xenograft tumors with APG350 and APG400 employing one treatment cycle with five consecutive daily intravenous injections showed a superior efficacy of APG350 with respect to tumor volume reduction and number of tumor free animals. Even mice with initially large tumors (∼600 mm3) were treated effectively with APG350. Remarkably, APG350 also showed pronounced dose dependent anti tumor efficacy on xenograft-tumors derived from colon cancer stem cells (CSC). Furthermore large CSC-derived tumors could be treated effectively and successful re-treatment of previously responding tumors demonstrated that tumors did not gain drug resistance. In a pilot experiment APG350 treatment of mice bearing a slowly growing primary rectum tumor xenograft also showed a significant tumor volume reduction. In all in vivo studies APG350 was well tolerated at doses between 0.3–100mg/kg bw in mice and doses up to 10mg/kg bw in a Cynomolgus monkey. General tolerability and potential effects on liver toxicity were assessed by co-application of APG350 together with a crosslinking antibody in mice. This treatment evoked only minor clinical signs at high doses with no relevant increase in liver enzymes. Currently two non-GLP toxicology studies, a 4-week study in mice and a dose escalation study in Cynomolgus monkeys are ongoing to confirm tolerability in vivo.
Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr LB-232. doi:10.1158/1538-7445.AM2011-LB-232