Abstract 935: Generation and preclinical characterization of CD123-CPI antibody-drug conjugate (ADC)

CD123 is expressed on cancer cells in a variety of hematologic malignancies including acute myeloid leukemia (AML). CD123 is frequently expressed on leukemic blasts and leukemic stem cells (LSCs), a cell population associated with relapse in patients, but there is minimal or no expression on most normal hematopoietic cells and solid tissues. Thus, CD123 is a promising target for AML. We have generated an anti-CD123 ADC that carries cyclopropylpyrroloindoline (CPI) payload that crosslinks DNA. A drug loading of 2 molecules of CPI per molecule of antibody was achieved by site-specific conjugation using our transglutaminase methodology. Upon binding to the CD123 antigen, CD123-ADC is internalized and delivered to the endosomal-lysosomal pathway whereupon the CPI payload is released from the antibody by proteolysis of the linker. The released CPI alkylates DNA, which activates ATR/ATM, CHK1, CHK2 and FANCD2, ultimately resulting in cell death. In vitro, CD123–CPI elicited cytotoxicity in a dose-dependent manner against several CD123-positive, but not against CD123-negative cell lines. Cell lines with higher CD123 expression level were more sensitive to the ADC. Long-Term Culture-Initiating Cells in vitro assay showed that AML patient bone marrow samples that naturally have high percentage of LSCs yielded substantially fewer colonies in CD123-ADC treated cells compared to control ADC. Importantly, CD123-ADC had no adverse effects in healthy donor bone marrow cells experimentally enriched in CD34+ primitive stem cells. This result suggests that CD123-ADC specifically inhibits the growth potential of leukemic blasts and progenitor cells. In vivo, robust antileukemic activity was observed in CD123-positive AML cell line-derived xenograft models. Low doses of CD123-ADC effectively regressed tumors whereas the tumor progressed in mice that received control ADC. Efficacy was also evaluated in disseminated AML PDX models (n≥7) established with patient samples of various cytogenetics and molecular abnormalities, and relapse/refractory. Flow cytometry analyses of leukemic load in peripheral blood and in bone marrow samples of mice showed that CD123-ADC was efficacious in reducing tumor burden even at the suboptimal doses. In summary, our data demonstrate that our CD123-ADC is highly active in a broad panel of primary AML samples. Currently, CD123-ADC in combination with other AML therapies is being tested in vivo. All these attributes of CD123-ADC make it an attractive agent to evaluate in clinical trials. Citation Format: Yoon-Chi Han, Fan Jiang, Nicole Piche-Nicholas, Madan Katragadda, Nadira Prashad, Manoj Charati, Wendy Hu, Mauricio Leal, Nathan Tumey, Andreas Maderna, Russell Dushin, Kenny Kim, LuAnna Lemon, Marc Damelin, HP Gerber, Lioudmila Tchistiakova, Frank Loganza, Chris O9Donnell, Puja Sapra. Generation and preclinical characterization of CD123-CPI antibody-drug conjugate (ADC) [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 935.