title: Improved circulating half-life and efficacy of an antibody-interleukin 2 immunocytokine based on reduced intracellular proteolysis.
abstract: OBJECTIVE
  Previous studies showed that reducing the interaction of antibody-interleukin 2 immunocytokines with Fc receptors improved their circulating half-life in mice and increased their antitumor activity. We sought to modify sequences that would increase half-life but retain the ability to activate Fc receptor-mediated effector functions.
  
  
  METHODS
  Modified immunocytokines were assessed in vitro for effector function and protease sensitivity and in vivo for pharmacokinetic and antitumor activities in an syngeneic tumor regression model.
  
  
  RESULTS
  Single amino acid changes in the junction sequence between the antibody and interleukin-2 components had dramatic effects on circulating half-life and antitumor activity. This effect was independent of Fc receptor binding to either Fcgamma receptors or the Fc protection receptor, but was associated with changes in susceptibility to intracellular proteases.
  
  
  CONCLUSIONS
  We have identified modifications that dramatically improve the circulating half-life of immunocytokines based on increased resistance to intracellular degradation and thus have demonstrated how these molecules can be recycled in and out of an intracellular compartment. Use of these improved immunocytokines with highly favorable pharmacokinetic properties and retained effector functions should lead to more effective treatment of epithelial cancers.
authors.affiliation: Lexigen Pharmaceuticals Corp., Lexington, Massachusetts 02421-3125, USA.
authors.forename: Stephen D
authors.initials: SD
authors.lastname: Gillies
authors.forename: Kin-Ming
authors.initials: KM
authors.lastname: Lo
authors.forename: Christa
authors.initials: C
authors.lastname: Burger
authors.forename: Yan
authors.initials: Y
authors.lastname: Lan
authors.forename: Thomas
authors.initials: T
authors.lastname: Dahl
authors.forename: Wai-Keung
authors.initials: WK
authors.lastname: Wong
mesh_terms: D000818:Animals; D056668:Cathepsin L; D002403:Cathepsins / Q000378:metabolism; D003110:Colonic Neoplasms / Q000276:immunology / Q000517:prevention & control; D003546:Cysteine Endopeptidases / Q000378:metabolism*; D006207:Half-Life; D006801:Humans; D018796:Immunoconjugates / Q000493:pharmacokinetics; D007141:Immunoglobulin Fc Fragments / Q000276:immunology; D007376:Interleukin-2 / Q000276:immunology* / Q000493:pharmacokinetics* / Q000627:therapeutic use; D051379:Mice; D008807:Mice, Inbred BALB C; D008822:Mice, Transgenic; D016297:Mutagenesis, Site-Directed; D009154:Mutation; D011993:Recombinant Fusion Proteins / Q000493:pharmacokinetics* / Q000627:therapeutic use; D014018:Tissue Distribution; D014162:Transfection; D014183:Transplantation, Heterologous; D014407:Tumor Cells, Cultured
chemical_list: D018796:Immunoconjugates; D007141:Immunoglobulin Fc Fragments; D007376:Interleukin-2; D011993:Recombinant Fusion Proteins; D002403:Cathepsins; D003546:Cysteine Endopeptidases; C534281:CTSL protein, human; D056668:Cathepsin L; C534278:Ctsl protein, mouse