A beta-lactamase with reduced immunogenicity for the targeted delivery of chemotherapeutics using antibody-directed enzyme prodrug therapy.

نویسندگان

  • Fiona A Harding
  • Amy D Liu
  • Marcia Stickler
  • O Jennifer Razo
  • Regina Chin
  • Nargol Faravashi
  • Wendy Viola
  • Tom Graycar
  • V Pete Yeung
  • Wolfgang Aehle
  • Daan Meijer
  • Stephanie Wong
  • M Harunur Rashid
  • Ana M Valdes
  • Volker Schellenberger
چکیده

Antibody-directed enzyme prodrug therapy (ADEPT) delivers chemotherapeutic agents in high concentration to tumor tissue while minimizing systemic drug exposure. beta-Lactamases are particularly useful enzymes for ADEPT systems due to their unique substrate specificity that allows the activation of a variety of lactam-based prodrugs with minimal interference from mammalian enzymes. We evaluated the amino acid sequence of beta-lactamase from Enterobacter cloacae for the presence of human T-cell epitopes using a cell-based proliferation assay using samples from 65 community donors. We observed a low background response that is consistent with a lack of preexposure to this enzyme. beta-Lactamase was found to contain four CD4+ T-cell epitopes. For two of these epitopes, we identified single amino acid changes that result in significantly reduced proliferative responses while retaining stability and activity of the enzyme. The beta-lactamase variant containing both changes induces significantly less proliferation in human and mouse cell assays, and 5-fold lower levels of IgG1 in mice were observed after repeat administration of beta-lactamase variant with adjuvant. The beta-lactamase variant should be very suitable for the construction of ADEPT fusion proteins, as it combines high activity toward lactam prodrugs, high plasma stability, a monomeric architecture, and a relatively low risk of eliciting an immune response in patients.

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عنوان ژورنال:
  • Molecular cancer therapeutics

دوره 4 11  شماره 

صفحات  -

تاریخ انتشار 2005