8081 Background: Ocaratzumab, previously known as AME-133v, is a humanized next-generation anti-CD20 monoclonal antibody. It has been optimized with a 13 to 20-fold increase in binding affinity to CD20 and improved binding to the low-affinity (F/F and F/V) polymorphisms of FcγRIIIa (CD16), which are thought to predict lower response rates and shorter duration of responses to rituximab. Methods: In a phase I dose escalation study in relapsed follicular lymphoma (FL) patients, ocaratuzumab was well-tolerated at doses up to 375 mg/m2 (Forero-Torres et al. CCR 2012). In a follow-on phase II trial, 44 patients with relapsed FL following prior rituximab and the low-affinity FcγRIIIa polymorphism (F-carriers) received 375 mg/m2 of ocaratuzumab weekly for 4 doses. In this study, overall response rate (ORR) was 36% and median progression free survival (PFS) was 91 weeks (Ganjoo et al. Haematologica 2011). Results: Amongst the 56 patients receiving 100 and 375 mg/m2 of ocaratuzumab, 8 patients had a previous time to progression of ≤ 180 days following their last rituximab treatment. These patients had a median of 2 prior rituximab treatments, (range 1-6 treatments), and median PFS following last treatment of 159 days. Five of the 8 patients showed a longer PFS after ocaratuzumab administration, compared with last rituximab treatment. All 5 patients expressed the homozygous low-affinity genotype of FcγRIIIa (F/F). At the time of study closure, 3 of the patients were still in remission (indicated by * in the table). Conclusions: This retrospective analysis suggests that ocaratuzumab may be non-cross-resistant to rituximab in patients with the low-affinity FcγRIIIa polymorphism. Prolonged PFS in selected patients following ocaratuzumab suggests that the increased binding affinity to CD16 and improved antibody-dependent cell-mediated cytotoxicity (ADCC) of this antibody is clinically relevant. As a single agent, ocaratuzumab may provide prolonged clinical benefit in relapsed FL patients and a clinical trial comparing ocaratuzumab to rituximab is in preparation. [Table: see text]

Purpose The efficacy of AME-133v in depleting B lymphocytes was compared in vitro to that of Rituxan, and the effect of FcαRIII genotype was assessed. Methods Blood samples from 37 healthy donors were collected and genotyped for FcαRIII polymorphism. Varying concentrations of AME-133v or Rituxan were applied to samples and CD19+ peripheral circulating B lymphocytes were measured using fluorescence-activated cell sorting (FACS). B cell depletion was listed as a percentage of baseline level. Results In an in vitro whole blood assay, AME-133v was more effective in depleting B lymphocytes from baseline levels than Rituxan, particularly at increasing concentrations (Table 1). Significant differences were found between treatments averaged over doses, between doses averaged over treatments, and between treatments depending on dose, all with p<0.0001. AME-133v's superior effect in depleting B cells is independent of dose (p<0.001), but caused greater B cell depletion at lower concentrations of antibody. In both AME-133v and Rituxan treatments, VF and FF patients demonstrated less B cell depletion, corroborating previous data citing decreased response to Rituxan among F-carriers and indicating improved pharmacodynamics of AME-133v. Conclusion AME-133v is engineered for a 6-fold more potent effector function in antibody-dependent cell-mediated cytotoxicity (ADCC) compared to Rituxan, which is elucidated through improved B cell depletion, across doses. Depletion in AME-133v-treated patients was also greater across all genotypes, suggesting superior pharmacodynamics of AME-133v in the low-responding F-carrier population. These findings indicate relevant biological activity of the antibody and suggest that AME-133v may provide additional clinical benefit for low-affinity FcαRIII patients. A randomized trial is being planned to compare clinical efficacy of AME-133v vs. Rituxan, targeting low-affinity FcαRIII patients. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 3784. doi:1538-7445.AM2012-3784

4977 Sustained Depletion of B-Cells by a Humanized, Fc-Engineered Anti-CD20 Antibody, AME-133v, in Patients with Relapsed Follicular Lymphoma J Wayne,1 K Ganjoo,2 A Forero,3 B Pohlman,4 S de Vos,5 S Carpenter,6 J Wooldridge,6 S Marulappa,1 V Jain11Mentrik Biotech, LLC, Dallas, TX, 2Standford University Medical Center, Stanford, CA, 3Comprehensive Cancer Center, University of Alabama at Birmingham, Birmingham, AL,4Cleveland Clinic Taussig Cancer Institute, Cleveland, OH, 5David Geffen School of Medicine at University of California, Los Angeles, CA, 6Eli Lilly and Company, Indianapolis, Indiana Introduction AME-133v is a humanized anti-CD20 monoclonal antibody that has a 13 to 20-fold increase in binding affinity and approximately 6-fold more potent effector function in antibody-dependent cell-mediated cytotoxicity (ADCC) compared to rituximab. Phase I/II clinical trials of AME-133v in patients with relapsed follicular lymphoma have demonstrated an overall response rate of greater than 30% with a complete response rate of 16%. The extent and duration of depletion of CD19+ B-cells in peripheral blood was used as a surrogate of therapeutic levels of AME-133v. Analysis from the Phase I/II clinical trials is presented in this report. Methods CD-19 positive B-cells in peripheral blood were measured in 77 patients with relapsed follicular lymphoma enrolled in two phase I/II clinical trials of AME-133v. These studies assessed five different doses of AME-133v (from 2 mg/m2 to 375 mg/m2). AME-133v was administered intravenously four times at weekly intervals in both trials. Blood samples were taken at multiple time points throughout the trial and a central lab measured levels of circulating CD19+ B-cells using fluorescence-activated cell sorting (FACS). Results Excluding the four patients enrolled in the 2 mg/m2 dose cohort, depletion of peripheral B-cells occurred in all patients and was sustained over time (Table 1). Baseline levels of B-cell counts ranged from 4 × 103 to 1,187 × 103 cells/μL, with an average of 102 × 103 cells/μL and a median of 60 × 103 cells/μL. Within 24 hours of the first infusion, all patients had depletion of circulating B-cells; ninety-six percent of patients had less than 10 × 103 cells/μL and two patients had less than 20 × 103 cells/μL. Interestingly, AME-133v was effective at depleting B-cells even at doses as low as 7.5 mg/m2. To assess sustainability of B-cell depletion after four doses of AME-133v, CD19+ cell counts were evaluated at nine weeks after the fourth infusion and every three months thereafter. Complete depletion of CD19+ lymphocytes was sustained for nine weeks. At five months after the last infusion of AME-133v, nearly two-thirds of patients had no detectable circulating B-cells. Sustained B-cell depletion lasted for at least eight months following the last infusion in 63% of patients. Table 1. B-cell counts for all patients in 7.5, 30, 100 and 375 mg/m2 cohorts. Percentages are cumulative Time Point Cell Count (x 103 cells/μL) 0 < 1 2 to 10 11 to 30 31 to 50 < 100 Day 1 (24 hours after last infusion) 62 % 66 % 96 % 100 % 100% 100% Day 7 (day of infusion 2) 75% 80% 95% 97% 97% 98% Day 28 (1 week after last infusion) 78 % 87% 95% 98% 98% 100% Day 84 (9 weeks after last infusion) 78% 87% 91% 96% 96% 98% Day 174 (5 months after last infusion) 60% 60% 70% 86% 93% 100% Day 264 (8 months after last infusion) 26% 26% 41% 63% 81% 89% Day 354 (11 months after last infusion) 0% 0% 15% 40% 55% 80% DEMOGRAPHIC CHARACTERISTICS (EVALUABLE POPULATION) “\f C \l 1 Demographic and Disease Characteristics on evaluable population (N=30) Conclusion The rapid and sustained effect of AME-133v on B-cell depletion, even in low-affinity FcγRIIIa patients, indicates a potentially relevant biological activity of the antibody in treating B-cell non-Hodgkin lymphoma. Notably, this depletion occurred even at very low doses of drug administration and persisted over time. This may be related to its higher affinity for CD20, increased ADCC, or both. The sustained B-cell depletion may result in prolonged clinical response and might mitigate the need for maintenance therapy. A randomized trial is being planned to compare efficacy of AME-133v vs. rituximab. Disclosures: No relevant conflicts of interest to declare.

2475 Introduction: Anti-CD20 antibodies like rituximab are traditionally administered by the intravenous routes although clinical trials by subcutaneous routes have been initiated. AME-133 is a monoclonal anti-CD20 antibody, has been engineered to be more potent than rituximab. It has 10–20 fold higher binding affinity for the CD20 epitope and approximately 6-fold greater potency in ADCC assays relative to rituximab. The higher potency may provide benefit to patients with low affinity FcgIIIRa and enable subcutaneous administration (patient convenience). In addition, AME-133 has been engineered to be potentially safer than rituximab; AME-133 is humanized to reduce immunogenicity and the CDC activity has been diminished to potentially reduce side effects associated with tumor lysis syndrome. Phase I/II clinical studies of AME-133 in rituximab pretreated/relapsed, patients with low affinity FcγIIIRa, follicular non-Hodgkin's lymphoma (NHL) in the US and in Japan have shown excellent safety profile and response rate (RR) greater than 30%. As a prelude to investigating the clinical efficacy and safety of AME-133 in subcutaneous formulation, a safety/PK study was conducted in cynomolgus monkeys. Objective: The objective of this study was to evaluate the systemic toxicity and pharmacokinetics of AME-133 in cynomolgus monkeys at three different dose levels when administered subcutaneously once each week for 14 consecutive weeks. In addition, the effect of AME-133 on B-cell depletion was evaluated. Study Design: A total of 48 naive cynomolgus monkeys (24 male and 24 female) were randomly assigned to four dose groups of AME-133 administered subcutaneously once weekly (0.0 mg/kg (vehicle), 0.6 mg/kg, 1.9 mg/kg and 6.0 mg/kg). Within the dose groups, there were two (6 week and 14 week) dosing schedules. In the 6 week dosing schedule, two-third of animals were sacrificed immediately after 6 weeks, and one-third of animals were sacrificed after 4 to 8 weeks of recovery period. In the 14 week dosing schedule, two-third of animals were sacrificed immediately after 14 weeks, and one-third of animals were sacrificed after 4 to 8 weeks of recovery period. Results: There was no AME-133 related toxicity. All dose levels of AME-133 substantially depleted B-cells in peripheral blood. The extent and duration of B-cell depletion was dose-dependent (Figure 1). B-cell depletion was not affected by gender of the animal. All dose levels of AME-133 substantially depleted the B-cells in peripheral blood beginning with the first post-dose time point (4 hours after infusion). The mid and high doses resulted in delayed return to normal during recovery in cynomolgus monkeys in 14 week dosing schedule. Conclusion: This study in cynomolgus monkeys showed AME-133 caused no toxicity, well absorbed and rapidly depleted circulating B-cells when administered subcutaneously. The No Observed Adverse Effect Level (NOAEL) is 6.0 mg/kg, the highest administered dose. These preliminary results justify further investigation of AME-133v administered subcutaneously in follicular lymphoma and other B-cell malignancies. Disclosures: No relevant conflicts of interest to declare.