Abstract 5662: JNJ-67571244: A novel anti-CD33 C2 domain binding bispecific antibody with potent T cell redirection activity
Authors: Nair-Gupta Priyanka , Diem Michael , Reeves Dara , Wang Weirong , Schulingkamp Robert , Sproesser Katrin , Mattson Bethany , Heidrich Bradley , Joseph Jocelin , Sendecki Jocelyn , Foulk Brad , Chu Gerald , Fink Damien , Jiao Qun , Wu Sheng-Jiun , Packman Kathryn , Elsayed Yusri , Attar Ricardo , Gaudet Francois 
Published: September 2020

Abstract
CD33 is expressed in 90% of acute myeloid leukemia (AML) patients with expression in both blasts and leukemic stem cells. The extracellular portion of CD33 consists of a V and a C2 Ig-like domain. A recent study showed that a single nucleotide polymorphism (SNP) rs12459419 (C &gt;T; Ala14Val in exon 2 of CD33) was present in ~50% of the Western AML population and is associated with preferential expression of an alternatively spliced CD33 isoform lacking exon2, resulting in the deletion of the CD33 V domain. Interestingly, several CD33-antibody-based therapies, including gemtuzumab ozogamicin (GO), the only approved anti-CD33 antibody drug conjugate for AML, bind and recognize the V domain of CD33. Recent data demonstrated that patients with SNP rs12459419 CC genotype receiving GO had a significantly lower risk of relapse and increased event-free survival compared to patients with the CT or TT genotypes. Given the data with GO, it is reasonable to hypothesize that the activity of other V binding CD33 antibodies maybe limited to a subset (~50%, rs12459419 CC genotype) of AML patients. On the other hand, since the C2 domain is shared by all CD33 isoforms in AML patients, we hypothesized that a C2 binding anti-CD33 antibody could target AML cells more broadly regardless of their SNP status. Additionally, we reasoned that targeting the membrane proximal C2 domain would be a beneficial strategy for a CD3 redirection bispecific antibody as targeting epitopes closer to the membrane have been reported to mediate efficient synapse formation between T cells and target cells leading to potent anti-tumor responses. We developed JNJ-67571244, a novel human bispecific antibody capable of binding to the C2 domain of CD33, and to CD3 to induce T-cell recruitment and tumor cell cytotoxicity independently of their SNP status. JNJ-67571244 specifically binds to CD33-expressing target cells and induces cytotoxicity of CD33+ AML cell lines in vitro at 48 hours (EC50 values: KG-1=0.168 nM, MOLM-13=0.131 nM, Kasumi-1=0.05 nM and OCI-AML3=0.183 nM) with concomitant T cell activation (EC50 values: KG-1=0.066 nM, MOLM-13=0.028 nM, Kasumi-1=0.043 nM and OCI-AML3=0.05 nM) along with cytokine release. In contrast, JNJ-67571244 was unable to kill CD33- cancer cell lines (CARNAVAL and KG-1 cells with a genetic deletion of CD33), demonstrating the specificity of the cytotoxicity. JNJ-67571244 demonstrated statistically significant anti-tumor activity in vivo in established disseminated and subcutaneous mouse models of human AML (MOLM-13Luc and KG-1: up to 100% and 92% tumor growth inhibition respectively) through T cell redirection activity. Furthermore, this antibody could deplete CD33+ blasts (EC50=0.549 nM) in AML patient blood samples (n=7) in an ex-vivo assay at 48 hours with concurrent T cell activation (EC50=0.355 nM). JNJ-67571244 also cross-reacts with cyno CD33 and CD3 and was well-tolerated in cynomolgus monkeys up to 30 mg/kg along with a sustained reduction in CD33+ leukocyte populations. Lastly, JNJ-67571244 mediated efficient cytotoxicity of cell lines and primary samples regardless of their genotype status (SNP rs12459419 CC, CT and TT), suggesting a potential therapeutic advantage over competitor V-binding antibodies. JNJ-67571244 is currently in Phase 1 clinical trials to treat relapsed/refractory AML and high risk myelodysplastic syndrome (MDS) patients (NCT03915379).
Citation Format: Priyanka Nair-Gupta, Michael Diem, Dara Reeves, Weirong Wang, Robert Schulingkamp, Katrin Sproesser, Bethany Mattson, Bradley Heidrich, Jocelin Joseph, Jocelyn Sendecki, Brad Foulk, Gerald Chu, Damien Fink, Qun Jiao, Sheng-Jiun Wu, Kathryn Packman, Yusri Elsayed, Ricardo Attar, Francois Gaudet. JNJ-67571244: A novel anti-CD33 C2 domain binding bispecific antibody with potent T cell redirection activity [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 5662.