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1055 Heterodimeric Fc-fused IL12 shows potent antitumor activity and low toxicity
  1. Wenting Liu1,
  2. Dayan Zhang1,
  3. Pengfei Zhou1,
  4. Guodong Shen2,
  5. Xiaoli Zeng3,
  6. Weiming Zhou1 and
  7. Liansheng Cheng1
  1. 1Hefei Hankemab Biotechnology CO., LTD, Anhui, China
  2. 2The First Affiliated Hospital of University of Science and Technology of China, Anhui, China
  3. 3Hefei Hankemab Biotechnology Co.,LTD, Hefei City, Shushan Distric, Hong Kong
  • Journal for ImmunoTherapy of Cancer (JITC) preprint. The copyright holder for this preprint are the authors/funders, who have granted JITC permission to display the preprint. All rights reserved. No reuse allowed without permission.


Background Interleukin-12 (IL12) is a heterodimeric pro-inflammatory cytokine (70 kDa) composed of two different polypeptide chains: an α-chain (p35 subunit) and a β-chain (p40 subunit). IL12 could induce the proliferation and cytotoxicity of T and NK cell and production of interferon-γ (IFN-γ), promote the differentiation of T helper 1 (TH1) cells and regulate innate resistance and adaptive immunity. However, the clinical efficacy of IL12 has been hindered for its high dose-related toxicities and short serum half-life. Here we have developed heterodimeric Fc-fused IL12, HK054-Vx, with reduced potency to improve tolerability and prolong half-life.

Methods We assessed the binding, bioactivity, anti-tumor efficacy and safety of HK054-Vx. The binding ability of IL12-Fc lead variants to activated PBMCs was analyzed by flow cytometry. The bioactivity of HK054-Vx was determined by assessing the activated T-cell proliferation and IFN-γ production in human PBMCs, and STAT-4 phosphorylation in a reporter gene assay.

Murine surrogates was engineered to mimic HK054-Vx to evaluate the antitumor efficacy in MC38 and CT26 syngeneic tumor mouse models. In vivo anti-tumor activity was also assessed by engrafting MCF-7 cancer cells into human PBMC engrafted BNDG-B2m mice. Cytokine release assay was conducted by incubating HK054-Vx with peripheral blood mononuclear cells (PBMCs) from healthy donors.

Results Heterodimeric Fc-fused IL12, HK054-Vx, were designed with reduced potency. HK054-Vx were observed to stimulate the proliferation of activated T cells and NK cells and induce production of IFN-γ in a dose-dependent manner. HK054-Vx could activate pSTAT4 signal in luciferase reporter gene assay and showed up to 100-fold reduction of potency compared to wild-type IL12-Fc. HK054-Vx showed potent anti-tumor efficacy in human PBMC engrafted BNDG-B2m mice and was better tolerated than WT mIL12-Fc.

Conclusions These data demonstrate that HK054-Vx, heterodimeric Fc-fused IL12 with reduced potency, retain strong anti-tumor efficacy with a favorable tolerability profile, which may provide a practical alternative to the systemic administration of IL12 for antitumor therapy.

This is an open access article distributed in accordance with the Creative Commons Attribution Non Commercial (CC BY-NC 4.0) license, which permits others to distribute, remix, adapt, build upon this work non-commercially, and license their derivative works on different terms, provided the original work is properly cited, appropriate credit is given, any changes made indicated, and the use is non-commercial. See

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