TNF-Selectokine: a novel prodrug generated for tumor targeting and site-specific activation of tumor necrosis factor

Oncogene. 2002 Jun 20;21(27):4257-65. doi: 10.1038/sj.onc.1205193.

Abstract

We describe a TNF fusion protein designated TNF-Selectokine, which is a homo-trimeric molecule comprised of a single chain antibody (scFv) targeting module, a trimerization domain and TNF. TNF-Selectokine exerts high bioactivity towards the targeted and adjacent, antigen negative cells. Membrane targeting dependent immobilization of the TNF-Selectokine induced cell death in TNFR1 and TNFR2 dependent manner, thus cell bound TNF-Selectokine mimicks membrane TNF. To restrict TNF activity to the tumor, a prototype of a TNF-Selectokine prodrug was constructed by insertion of a TNFR1 fragment, separated from TNF by a protease-sensitive linker. The prodrug exerts minimal TNF activity, but can be activated in vitro several thousand-fold by proteolytic digest, showing the principal feasibility of this approach. Choice of cleavage site(s) recognized by protease(s) typically associated with a given carcinoma should allow high dose systemic application of the respective TNF prodrug that unveils its specific bioactivity only in targeted tissues.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adenocarcinoma / pathology
  • Amino Acid Sequence
  • Animals
  • Antigen-Antibody Reactions
  • Antigens, CD / drug effects
  • Antigens, CD / genetics
  • Antineoplastic Agents / chemistry
  • Antineoplastic Agents / metabolism
  • Antineoplastic Agents / pharmacology*
  • Apoptosis / drug effects
  • Binding Sites
  • Biotransformation
  • CHO Cells
  • Coculture Techniques
  • Colonic Neoplasms / pathology
  • Cricetinae
  • Cricetulus
  • Drug Design
  • Humans
  • Immunoglobulin Fragments / chemistry
  • Immunoglobulin Fragments / genetics
  • Immunoglobulin Fragments / metabolism
  • Immunoglobulin Fragments / pharmacology*
  • Models, Molecular
  • Molecular Sequence Data
  • Prodrugs / metabolism
  • Prodrugs / pharmacology*
  • Protein Conformation
  • Protein Structure, Tertiary
  • Receptors, Tumor Necrosis Factor / drug effects
  • Receptors, Tumor Necrosis Factor / genetics
  • Receptors, Tumor Necrosis Factor, Type I
  • Receptors, Tumor Necrosis Factor, Type II
  • Recombinant Fusion Proteins / genetics
  • Recombinant Fusion Proteins / metabolism
  • Recombinant Fusion Proteins / pharmacology
  • Rhabdomyosarcoma / pathology
  • Single-Chain Antibodies
  • Tenascin / chemistry
  • Tenascin / genetics
  • Trypsin / metabolism
  • Trypsin / pharmacology
  • Tumor Cells, Cultured / metabolism
  • Tumor Cells, Cultured / pathology
  • Tumor Necrosis Factor-alpha / chemistry
  • Tumor Necrosis Factor-alpha / metabolism*
  • Tumor Necrosis Factor-alpha / pharmacology*

Substances

  • Antigens, CD
  • Antineoplastic Agents
  • Immunoglobulin Fragments
  • Prodrugs
  • Receptors, Tumor Necrosis Factor
  • Receptors, Tumor Necrosis Factor, Type I
  • Receptors, Tumor Necrosis Factor, Type II
  • Recombinant Fusion Proteins
  • Single-Chain Antibodies
  • TNF-selectokine
  • Tenascin
  • Tumor Necrosis Factor-alpha
  • Trypsin