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Enhancement of tumor necrosis factor α antitumor immunotherapeutic properties by targeted delivery to aminopeptidase N (CD13)

Nature Biotechnology volume 18, pages 1185–1190 (2000)Cite this article

Abstract

The clinical use of tumor necrosis factor α (TNF) as an anticancer drug is limited to local treatments because of its dose-limiting systemic toxicity. We show here that murine TNF fused with CNGRC peptide (NGR-TNF), an aminopeptidase N (CD13) ligand that targets activated blood vessels in tumors, is 12–15 times more efficient than murine TNF in decreasing the tumor burden in lymphoma and melanoma animal models, whereas its toxicity is similar. Similarly, human NGR-TNF induced stronger antitumor effects than human TNF, even with 30 times lower doses. Coadministration of murine NGR-TNF with a CNGRC peptide or an anti-CD13 antibody markedly decreased its antitumor effects. Tumor regression, induced by doses of murine NGR-TNF lower than the LD50, was accompanied by protective immunity. In contrast, no cure was induced by TNF at any dose. These results suggest that targeted delivery of TNF to CD13 may enhance its immunotherapeutic properties. Moreover, these findings reveal the potential of tumor homing peptides to generate a new class of recombinant cytokines that compared to immunocytokines have a simpler structure, could be easier to produce and are potentially less immunogenic.

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Figure 1: Effect of murine TNF and NGR-TNF on the tumor growth and body weight of animals bearing RMA-T lymphomas (A,B,D,E) or B16F1 melanomas (C,F).
Figure 2: Effect of murine TNF and NGR-TNF, administered in combination with melphalan, on the growth of B16F1 melanoma.
Figure 3: Effect of limited tryptic digestion of murine NGR-TNF and TNF on their mass (A) and antitumor activity (B).
Figure 4: Effect of mAb R3-63 and CNGRC on the antitumor activity of murine NGR-TNF and TNF.
Figure 5: Cytotoxic activity of murine NGR-TNF and TNF against CD13-positive HL60 (C) and U937 cells (F).
Figure 6: SDS–PAGE and antitumor activity of human NGR-TNF before and after denaturation–refolding.

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Acknowledgements

This work was supported by Associazione Italiana Ricerca sul Cancro (AIRC). We thank Renata Pasqualini for advice and critical review, Barbara Colombo for the protein characterization and endotoxin analysis, and A.G. Siccardi and P. Dellabona for helpful discussions.

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  1. Department of Biological and Technological Research San Raffaele H Scientific Institute, via Olgettina 58, 20132, Milan, Italy

    Flavio Curnis, Angelina Sacchi, Laura Borgna, Fulvio Magni, Anna Gasparri & Angelo Corti

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Correspondence to Angelo Corti.

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Curnis, F., Sacchi, A., Borgna, L. et al. Enhancement of tumor necrosis factor α antitumor immunotherapeutic properties by targeted delivery to aminopeptidase N (CD13). Nat Biotechnol 18, 1185–1190 (2000). https://doi.org/10.1038/81183

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