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Immunotoxin pharmacokinetics: a comparison of the anti-glioblastoma bi-specific fusion protein (DTAT13) to DTAT and DTIL13

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Summary

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.

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Acknowledgement

This work was supported in part by funding from the Peyton Society, Martha L. Kramer Fund, and Funds from the NCI, NIH RO1 CA 108637.

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Authors and Affiliations

  1. Departments of Neurosurgery, University of Minnesota Cancer Center, Minneapolis, MN, USA

    Edward Rustamzadeh, Walter C. Low & Walter A. Hall

  2. Departments of Pediatrics, University of Minnesota Cancer Center, Minneapolis, MN, USA

    Daniel A. Vallera & Deborah A. Todhunter

  3. Departments of Therapeutic Radiology-Radiation Oncology, University of Minnesota Cancer Center, Minneapolis, MN, USA

    Angela Panoskaltsis-Mortari

  4. Biophysical Sciences and Medical Physics Doctoral Program, University of Minnesota, Minneapolis, MN, USA

    Edward Rustamzadeh

  5. University of Minnesota Cancer Center, Mayo Mail Code: 367, Harvard St. at East River Road, Minneapolis, MN, 55455, USA

    Daniel A. Vallera

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  1. Edward Rustamzadeh
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  2. Daniel A. Vallera
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  3. Deborah A. Todhunter
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  4. Walter C. Low
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  5. Angela Panoskaltsis-Mortari
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  6. Walter A. Hall
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Correspondence to Daniel A. Vallera.

Additional information

This work was supported in part by the US Public Health Service Grants RO1-CA1-108637 awarded by the NCI and the NIAID, DHHS and Children’s Cancer Research Fund.

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Rustamzadeh, E., Vallera, D.A., Todhunter, D.A. et al. Immunotoxin pharmacokinetics: a comparison of the anti-glioblastoma bi-specific fusion protein (DTAT13) to DTAT and DTIL13. J Neurooncol 77, 257–266 (2006). https://doi.org/10.1007/s11060-005-9051-7

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