The Antibody-Based Delivery of Interleukin-12 to the Tumor Neovasculature Eradicates Murine Models of Cancer in Combination with Paclitaxel

2013

Intl Journal of Cancer

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Preclinical studies with recombinant murine interleukin 4 (IL4) in models of cancer have shown potent tumor growth inhibition. However, systemic administration of human IL4 to cancer patients exhibited modest antitumor activity and considerable toxicities. To improve the therapeutic index and reduce side effects of this cytokine, we developed of a novel "immunocytokine" based on sequential fusion of murine IL4 with the antibody fragment F8 (specific to the alternatively spliced extra-domain A of fibronectin, a marker for tumor-angiogenesis) in diabody format. The resulting fusion protein, termed F8-IL4, retained full antigen-binding activity and cytokine bioactivity and was able to selectively localize on solid tumors in vivo. When used as single agent, F8-IL4 inhibited tumor growth in three different immunocompetent murine cancer models (F9 teratocarcinoma, CT26 colon carcinoma and A20 lymphoma). Furthermore, F8-IL4 showed synergistic effects when coadministered with immunocytokines based on IL2 and IL12. Indeed, combination therapy with an IL12-based immunocytokine yielded complete tumor eradication, in spite of the fact that IL4 and IL12 display opposite immunological mechanisms of action in terms of their polarization of T-cell based responses. No weight loss or any signs of toxicity were observed in treated mice, both in monotherapy and in combination, indicating a good tolerability of the immunocytokine treatment. Interestingly, mice cured from CT26 tumors acquired a durable protective antitumor immunity. Depletion experiments indicated that the antitumor activity was mediated by CD81 T cells and by NK cells.Cytokines are potent modulators of the activity of the immune system, which can be used for therapeutic purposes. A number of cytokines (e.g., IL2, interferon-alpha and TNF) are frequently used for the treatment of cancer patients, but these agents can be toxic even at low doses (i.e., few milligrams), preventing the escalation to therapeutically active regimens.1 The antibody-based targeted delivery of cytokines ("immunocytokines") to the site of disease represents a promising avenue for increasing the therapeutic index of cytokine products, increasing activity and sparing normal organs.2 Proinflammatory immunocytokines (e.g., those based on IL2, IL12, IL15 and TNF) often display a potent antitumoral effect in mouse models of cancer.3-7 By contrast, antiinflammatory immunocytokines (e.g., those based on IL10) confer a therapeutic benefit in mouse models of chronic inflammatory conditions (rheumatoid arthritis and endometriosis 8,9 ) but have no impact on tumor growth (Trachsel and Neri, unpublished results).We have mainly used antibodies specific to splice-isoforms of fibronectin and of tenascin-C as vehicles for pharmacodelivery applications, as these antigens are virtually undetectable in the normal healthy adult (exception made for placenta, endometrium and some vessels in the ovaries) while being strongly expressed in the majority of solid tumors and lymphomas, with a prominent vascular staining p...

Section: Discussionmentioning

confidence: 99%

“…3,7,[35][36][37] We first performed combination experiments in F9 tumor-bearing mice using F8-IL4 plus F8-IL2, 5 in consideration of the fact that IL2-based products are the most frequently investigated immunocytokines in clinical trials. which led to complete tumor eradications in 2=5 mice.…”

Section: Production and In Vitro Characterization Of Il4-based Immunomentioning

confidence: 99%

The antibody‐based targeted delivery of interleukin‐4 and 12 to the tumor neovasculature eradicates tumors in three mouse models of cancer

Hemmerle

2013

Intl Journal of Cancer

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“…9), while they are expressed abundantly in the neovasculature and stroma of virtually all aggressive tumors in mice and humans (7,(10)(11)(12)(13). Some proinflammatory cytokines [e.g., interleukin (IL)2, IL12, and TNF] that were fused to L19 or F8 have exhibited impressive anticancer activity and selective uptake at the tumor site (11)(12)(13)(14)(15)(16)(17)(18)(19)(20). Other cytokines have shown limitations either in tumor targeting or in therapy experiments (e.g., IL7, IL17, IL15, and IL18; refs.…”

Section: Introductionmentioning

confidence: 99%

The Dose-Dependent Tumor Targeting of Antibody–IFNγ Fusion Proteins Reveals an Unexpected Receptor-Trapping Mechanism In Vivo

Hemmerle

Cancer Immunology Research

2014

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Cytokines often display substantial toxicities at low concentrations, preventing their escalation for therapeutic treatment of cancer. Fusion proteins comprising cytokines and recombinant antibodies may improve the anticancer activity of proinflammatory cytokines. Murine IFNg was appended in the diabody format at the C-terminus of the F8 antibody, generating the F8-IFNg fusion protein. The F8 antibody is specific for the extra-domain A (EDA) of fibronectin, a tumor-associated antigen that is expressed in the vasculature and stroma of almost all tumor types. Tumor-targeting properties were measured in vivo using a radioiodinated preparation of the fusion protein. Therapy experiments were performed in three syngeneic murine models of cancer [F9 teratocarcinoma, WEHI-164 fibrosarcoma, and Lewis lung carcinoma (LLC)]. F8-IFNg retained the biologic activity of both the antibody and the cytokine moiety in vitro, but, unlike the parental F8 antibody, it did not preferentially localize to the tumors in vivo. However, when unlabeled F8-IFNg was administered before radioiodinated F8-IFNg, a selective accumulation at the tumor site was observed. F8-IFNg showed dose-dependent anticancer activity with a clear superiority over untargeted recombinant IFNg. The anticancer activity was potentiated by combining with F8-IL4 without additional toxicities, whereas combination of F8-IFNg with F8-TNF was lethal in all mice. Unlike other antibodycytokine fusions, the use of IFNg as payload for anticancer therapy is associated with a receptor-trapping mechanism, which can be overcome by the administration of a sufficiently large amount of the fusion protein without any detectable toxicity at the doses used. Cancer Immunol Res; 2(6); 559-67. Ó2014 AACR.

“…members of the IL12 superfamily) can offer additional design possibilities, depending on the assembly of the two cytokine subunits (Fig. 1c) [40][41][42][43]. When considering IgG-based immunocytokines, C-terminal fusions to the heavy or to the light chains have been proposed [44].…”

Section: Immunocytokine Formats and Target Antigensmentioning

confidence: 99%

“…For more-complex cytokines (e.g. IL12) biodistribution properties heavily depend on the format chosen for the fusion with antibody moieties [42].…”

Section: Disease-homing Properties Of Immunocytokinesmentioning

confidence: 99%

Immunocytokines: a novel class of products for the treatment of chronic inflammation and autoimmune conditions

Bootz

2016

Drug Discovery Today

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Antibody-cytokine fusion proteins, often referred to as immunocytokines, represent a novel class of biopharmaceutical agents that combine the disease-homing activity of certain antibodies with the immunomodulatory properties of cytokine payloads. Originally, immunocytokines were mainly developed for cancer therapy applications. More recently, however, the use of antiinflammatory cytokines for the treatment of chronic inflammatory conditions and to treat autoimmune diseases has been considered. This review analyzes basic principles in the design of immunocytokines and describes the most advanced products in preclinical and clinical development.