Immunotoxin pharmacokinetics: a comparison of the anti-glioblastoma bi-specific fusion protein (DTAT13) to DTAT and DTIL13.
DTAT13, a novel recombinant bispecific immunotoxin (IT) consisting of truncated diphtheria toxin, an amino-terminal (AT) fragment of the urokinase-type plasminogen activator (uPa), and a fragment of human IL-13 was assembled in order to target receptors on glioblastoma multiforme (GBM) and its associated neovasculature. Previous in vitro studies confirmed the efficacy of DTAT13 against various GBM cell lines expressing both IL-13 receptor or uPA receptor, and previous in vivo testing demonstrated the efficacy of DTAT13 in significantly inhibiting a range of xenograft tumors and showed that DTAT13 was 160- and 8-fold less toxic to the parental fusion IT, DTAT and DTIL13, respectively. To further understand the properties of DTAT13, pharmacokinetic/biodistribution experiments were performed. Binding analysis revealed that the IL-13 domain functioned independently of the uPA domain and that the K (d) for each binding domain was essentially the same as that of DTIL13 and DTAT. Flow cytometry studies indicated that DTAT13 bound better than DTAT or DTIL13. Analysis of the rate of protein synthesis inhibition in U87 MG cells by DTAT13 compared to DTAT revealed a faster rate of inhibition with DTAT13 compared to DTAT. The rate of protein synthesis inhibition of DTAT13 was identical to that of DTIL13 in U373 MG cells. Intracranial biodistribution studies revealed that DTAT13 was able to cross to the contralateral hemisphere unlike DTIL13 but similar to DTAT. These studies show that DTAT13 has properties encompassing those of both DTIL13 and DTAT and warrants further consideration for clinical development.
Rustamzadeh
Edward
E
Department of Neurosurgery, University of Minnesota Cancer Center, Minneapolis, MN 55455, USA.
Vallera
Daniel A
DA
Todhunter
Deborah A
DA
Low
Walter C
WC
Panoskaltsis-Mortari
Angela
A
Hall
Walter A
WA
D000818:Animals; D001921:Brain / Q000378:metabolism; D004167:Diphtheria Toxin / Q000493:pharmacokinetics* / Q000627:therapeutic use; D005260:Female; D005909:Glioblastoma / Q000098:blood supply / Q000188:drug therapy / Q000276:immunology / Q000378:metabolism*; D006801:Humans; D000922:Immunotoxins / Q000493:pharmacokinetics* / Q000627:therapeutic use; D018793:Interleukin-13 / Q000493:pharmacokinetics* / Q000627:therapeutic use; D051379:Mice; D008810:Mice, Inbred C57BL; D009389:Neovascularization, Pathologic / Q000188:drug therapy / Q000276:immunology* / Q000378:metabolism; D014176:Protein Biosynthesis / Q000187:drug effects; D011993:Recombinant Fusion Proteins / Q000493:pharmacokinetics / Q000627:therapeutic use; D014568:Urokinase-Type Plasminogen Activator / Q000493:pharmacokinetics* / Q000627:therapeutic use
D004167:Diphtheria Toxin; D000922:Immunotoxins; D018793:Interleukin-13; D011993:Recombinant Fusion Proteins; C471586:diphtheria toxin-urokinase fusion protein, human; D014568:Urokinase-Type Plasminogen Activator