TNF-Selectokine: a novel prodrug generated for tumor targeting and site-specific activation of tumor necrosis factor.
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.
Wüest
Thomas
T
Institute of Cell Biology and Immunology, University of Stuttgart, Germany.
Gerlach
Elke
E
Banerjee
Debola
D
Gerspach
Jeannette
J
Moosmayer
Dieter
D
Pfizenmaier
Klaus
K
D000230:Adenocarcinoma / Q000473:pathology; D000595:Amino Acid Sequence; D000818:Animals; D000937:Antigen-Antibody Reactions; D015703:Antigens, CD / Q000187:drug effects / Q000235:genetics; D000970:Antineoplastic Agents / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology*; D017209:Apoptosis / Q000187:drug effects; D001665:Binding Sites; D001711:Biotransformation; D016466:CHO Cells; D018920:Coculture Techniques; D003110:Colonic Neoplasms / Q000473:pathology; D006224:Cricetinae; D003412:Cricetulus; D015195:Drug Design; D006801:Humans; D007128:Immunoglobulin Fragments / Q000737:chemistry / Q000235:genetics / Q000378:metabolism / Q000494:pharmacology*; D008958:Models, Molecular; D008969:Molecular Sequence Data; D011355:Prodrugs / Q000378:metabolism / Q000494:pharmacology*; D011487:Protein Conformation; D017434:Protein Structure, Tertiary; D018124:Receptors, Tumor Necrosis Factor / Q000187:drug effects / Q000235:genetics; D047888:Receptors, Tumor Necrosis Factor, Type I; D047889:Receptors, Tumor Necrosis Factor, Type II; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism / Q000494:pharmacology; D012208:Rhabdomyosarcoma / Q000473:pathology; D057127:Single-Chain Antibodies; D019063:Tenascin / Q000737:chemistry / Q000235:genetics; D014357:Trypsin / Q000378:metabolism / Q000494:pharmacology; D014407:Tumor Cells, Cultured / Q000378:metabolism / Q000473:pathology; D014409:Tumor Necrosis Factor-alpha / Q000737:chemistry / Q000378:metabolism* / Q000494:pharmacology*
D015703:Antigens, CD; D000970:Antineoplastic Agents; D007128:Immunoglobulin Fragments; D011355:Prodrugs; D018124:Receptors, Tumor Necrosis Factor; D047888:Receptors, Tumor Necrosis Factor, Type I; D047889:Receptors, Tumor Necrosis Factor, Type II; D011993:Recombinant Fusion Proteins; D057127:Single-Chain Antibodies; C460220:TNF-selectokine; D019063:Tenascin; D014409:Tumor Necrosis Factor-alpha; D014357:Trypsin