title: Synergistic therapeutic effects of a tumor targeting antibody fragment, fused to interleukin 12 and to tumor necrosis factor alpha.
abstract: The potent antitumor activity of certain cytokines is often achieved at the expense of unacceptable toxicity. One avenue to improve the therapeutic index of cytokines in cancer therapy consists of fusing them to monoclonal antibodies capable of a selective localization at the tumor site. We have constructed fusion proteins of interleukin-12 (IL-12) and tumor necrosis factor (TNF-alpha) with L19, an antibody fragment specific to the extradomain B of fibronectin which has been shown to target tumors in animal models and in patients with cancer. These fusions display a potent antitumor activity in several immunocompetent murine models of cancer but do not lead to complete remissions of established aggressive tumors. In this article, we have evaluated the tumor-targeting properties and the anticancer activities of combinations of the two antibody-cytokine fusion proteins, as well as of a triple fusion protein between IL-12, L19, and TNF-alpha. Although all fusion proteins were active in vitro, the triple fusion protein failed to localize to tumors in vivo and to show significant therapeutic effects. By contrast, the combination of IL-12-L19 and L19-TNF-alpha displayed potent synergistic anticancer activity and led to the eradication of F9 teratocarcinomas grafted in immunocompetent mice. When cured mice were rechallenged with tumor cells, a delayed onset of tumor growth was observed, indicating the induction of a partial antitumor vaccination effect. Potent anticancer effects were achieved at doses of IL-12-L19 and L19-TNF-alpha (2 micro g + 2 micro g/mouse), which were at least 5-fold lower than the maximal-tolerated dose. The combined administration of the two fusion proteins showed only a modest increase in toxicity, compared with treatments performed with the individual fusion proteins. These results show that the targeted delivery of cytokines to the tumor environment strongly potentiates their antitumor activity and that the combination treatment with IL-12-L19 and L19-TNF-alpha appears to be synergistic in vivo.
authors.affiliation: Institute of Pharmaceutical Sciences, Swiss Federal Institute of Technology Zurich, CH-8057 Zurich, Switzerland.
authors.forename: Cornelia
authors.initials: C
authors.lastname: Halin
authors.forename: Verena
authors.initials: V
authors.lastname: Gafner
authors.forename: Maria Elena
authors.initials: ME
authors.lastname: Villani
authors.forename: Laura
authors.initials: L
authors.lastname: Borsi
authors.forename: Alexander
authors.initials: A
authors.lastname: Berndt
authors.forename: Hartwig
authors.initials: H
authors.lastname: Kosmehl
authors.forename: Luciano
authors.initials: L
authors.lastname: Zardi
authors.forename: Dario
authors.initials: D
authors.lastname: Neri
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D016503:Drug Delivery Systems; D004354:Drug Screening Assays, Antitumor; D004357:Drug Synergism; D005260:Female; D018796:Immunoconjugates / Q000008:administration & dosage / Q000493:pharmacokinetics / Q000627:therapeutic use* / Q000633:toxicity; D007135:Immunoglobulin Variable Region / Q000008:administration & dosage / Q000627:therapeutic use* / Q000633:toxicity; D007167:Immunotherapy; D007371:Interferon-gamma / Q000378:metabolism; D018664:Interleukin-12 / Q000008:administration & dosage / Q000493:pharmacokinetics / Q000627:therapeutic use* / Q000633:toxicity; D051379:Mice; D009368:Neoplasm Transplantation; D011993:Recombinant Fusion Proteins / Q000008:administration & dosage / Q000627:therapeutic use / Q000633:toxicity; D013601:T-Lymphocytes / Q000378:metabolism; D018243:Teratocarcinoma / Q000473:pathology / Q000628:therapy*; D014018:Tissue Distribution; D014407:Tumor Cells, Cultured; D014409:Tumor Necrosis Factor-alpha / Q000008:administration & dosage / Q000493:pharmacokinetics / Q000627:therapeutic use* / Q000633:toxicity; D014611:Vaccination
chemical_list: D000911:Antibodies, Monoclonal; D018796:Immunoconjugates; D007135:Immunoglobulin Variable Region; D011993:Recombinant Fusion Proteins; D014409:Tumor Necrosis Factor-alpha; D018664:Interleukin-12; D007371:Interferon-gamma