Abstract 2473: A novel anti-PD-1 and proIL-2 bifunctional fusion protein delivers potent anti-tumor activity via PD-1 checkpoint inhibition and tumor selective IL-2R agonism preferentially on antigen specific T cells
Authors: Zijuan Li, Feifei Zhang, Ze Zhang, Harry Zhou
Journal: Cancer Research (2024)

Abstract High-dose IL-2 (HD IL-2) induces complete and durable responses in RCC and melanoma but the treatment is associated with severe toxicities. PD-(L)1 blockade represents a recent breakthrough in immunotherapy with solid tumors, but only 20-30% of patients can benefit from the therapy and most responders eventually develop resistance. In more recent clinical studies with small number of subjects, HD IL-2 has shown promising responses in patients who progressed on anti-PD-1 therapy and combination therapy of HD-IL-2 and PD-1 blockade has resulted in high rate of objective response. In an attempt to maximize the clinical benefit of these immunotherapies to a broader patient population with deeper response and in the meantime to minimize the severe toxicity of HD IL-2, we have generated a novel bifunctional PD-1-proIL-2v fusion protein harnessing the mechanisms of PD-1 blockade and PD-1-cis directed tumor conditional IL-2R activation. PD-1-proIL-2v shows dose dependent inhibition of PD-1 immune checkpoint pathway measured by GM-CSF and IFNγ production in an MLR assay. Protease activated PD-1-proIL-2v, but not the intact drug, stimulates the phosphorylation of STAT5 of NK and T cells, the proliferation of CD8 T cells as well as the cytokine production in human whole blood. The potency of pSTAT5 by protease activated product is enhanced by 10-20 folds on PD-1+ T cells than T cells without PD-1. When tested in mouse tumor models resistant to PD-1 blockade, PD1-proIL-2v demonstrated high potency of tumor inhibition at as low as 0.1 mg/kg dose. In contrast, non-targeted proIL-2v at molar equivalent or higher dose showed minimal antitumor effect in either monotherapy or combination with anti-PD-1. Analysis of PD-1-proIL-2v treated mice showed dramatically increased tumor infiltrating CD4+ and CD8+ T cells, including PD-1+Tcf-1+ and PD-1+ IL-18Ra+TIM3− CD8+ T cells, referred to “stem-like” progenitor effector cells and “fresh” effector cells, respectively. Activation of the IL-2 procytokine in PD-1-proIL-2v was observed in mouse tumor tissues but not in the plasma. In monkey GLP-compliant 4-week repeat-dose toxicity study, PD-1-proIL-2v demonstrated excellent safety profiles without overt irAEs that are characteristic of IL-2. In conclusion, our preclinical data demonstrated that PD-1-proIL-2v effectively stimulated the expansion and reinvigoration of antigen specific T cells in the tumor tissue resulting in strong anti-tumor efficacy while it did not cause significant peripheral toxicity. Phase 1 clinical trial of PD-1-proIL-2v is planned to start in early 2024 to evaluate the safety, pharmacokinetics and anti-tumor activity in advanced solid tumors. Citation Format: Zijuan Li, Feifei Zhang, Ze Zhang, Harry Zhou. A novel anti-PD-1 and proIL-2 bifunctional fusion protein delivers potent anti-tumor activity via PD-1 checkpoint inhibition and tumor selective IL-2R agonism preferentially on antigen specific T cells [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 2473.