Target-selective activation of a TNF prodrug by urokinase-type plasminogen activator (uPA) mediated proteolytic processing at the cell surface

Cancer Immunol Immunother. 2006 Dec;55(12):1590-600. doi: 10.1007/s00262-006-0162-6. Epub 2006 Apr 25.

Abstract

We have previously developed TNF prodrugs comprised of a N-terminal scFv targeting, a TNF effector and a C-terminal TNFR1-derived inhibitor module linked to TNF via a MMP-2 motif containing peptide, allowing activation by MMP-2-expressing tumor cells. To overcome the known heterogeneity of matrix metalloprotease expression, we developed TNF prodrugs that become processed by other tumor and/or stroma-associated proteases. These TNF prodrugs comprise either an uPA-selective or a dual uPA-MMP-2-specific linker which displayed efficient, target-dependent and cleavage sequence-specific activation by the corresponding tumor cell-expressed proteases. Selective pharmacologic inhibition of endogenous uPA and MMP-2 confirm independent prodrug processing by these two model proteases and indicate the functional superiority of a prodrug containing a multi-specific protease linker. Processing optimised TNF prodrugs should increase the proportion of active therapeutic within the targeted tissue and thus potentially enhance tumor response rate.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antigens, Neoplasm / metabolism
  • Apoptosis
  • Biomarkers, Tumor / metabolism
  • Cell Line, Tumor
  • Cell Membrane / enzymology
  • Gelatinases
  • Humans
  • Matrix Metalloproteinase 2 / metabolism
  • Membrane Proteins
  • Peptides / chemistry
  • Peptides / metabolism
  • Prodrugs / chemistry
  • Prodrugs / metabolism*
  • Prodrugs / pharmacology
  • Receptors, Tumor Necrosis Factor, Type I / chemistry
  • Receptors, Tumor Necrosis Factor, Type I / metabolism*
  • Serine Endopeptidases / metabolism
  • Tumor Necrosis Factor-alpha / chemistry
  • Tumor Necrosis Factor-alpha / metabolism*
  • Tumor Necrosis Factor-alpha / pharmacology
  • Urokinase-Type Plasminogen Activator / metabolism*

Substances

  • Antigens, Neoplasm
  • Biomarkers, Tumor
  • Membrane Proteins
  • Peptides
  • Prodrugs
  • Receptors, Tumor Necrosis Factor, Type I
  • Tumor Necrosis Factor-alpha
  • Serine Endopeptidases
  • fibroblast activation protein alpha
  • Urokinase-Type Plasminogen Activator
  • Gelatinases
  • Matrix Metalloproteinase 2