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Investigational New Drugs

, Volume 34, Issue 2, pp 149–158 | Cite as

Preclinical toxicity evaluation of a novel immunotoxin, D2C7-(scdsFv)-PE38KDEL, administered via intracerebral convection-enhanced delivery in rats

  • Xuhui Bao
  • Vidyalakshmi Chandramohan
  • Randall P. Reynolds
  • John N. Norton
  • William C. Wetsel
  • Ramona M. Rodriguiz
  • Dipendra K. Aryal
  • Roger E. McLendon
  • Edward D. Levin
  • Neil A. Petry
  • Michael R. Zalutsky
  • Bruce K. Burnett
  • Chien-Tsun Kuan
  • Ira H. Pastan
  • Darell D. BignerEmail author
PRECLINICAL STUDIES

Summary

D2C7-(scdsFv)-PE38KDEL (D2C7-IT) is a novel immunotoxin that reacts with wild-type epidermal growth factor receptor (EGFRwt) and mutant EGFRvIII proteins overexpressed in glioblastomas. This study assessed the toxicity of intracerebral administration of D2C7-IT to support an initial Food and Drug Administration Investigational New Drug application. After the optimization of the formulation and administration, two cohorts (an acute and chronic cohort necropsied on study days 5 and 34) of Sprague–Dawley (SD) rats (four groups of 5 males and 5 females) were infused with the D2C7-IT formulation at total doses of 0, 0.05, 0.1, 0.4 μg (the acute cohort) and 0, 0.05, 0.1, 0.35 μg (the chronic cohort) for approximately 72 h by intracerebral convection-enhanced delivery using osmotic pumps. Mortality was observed in the 0.40 μg (5/10 rats) and 0.35 μg (4/10 rats) high-dose groups of each cohort. Body weight loss and abnormal behavior were only revealed in the rats treated with high doses of D2C7-IT. No dose-related effects were observed in clinical laboratory tests in either cohort. A gross pathologic examination of systemic tissues from the high-dose and control groups in both cohorts exhibited no dose-related or drug-related pathologic findings. Brain histopathology revealed the frequent occurrence of dose-related encephalomalacia, edema, and demyelination in the high-dose groups of both cohorts. In this study, the maximum tolerated dose of D2C7-IT was determined to be between 0.10 and 0.35 μg, and the no-observed-adverse-effect-level was 0.05 μg in SD rats. Both parameters were utilized to design the Phase I/II D2C7-IT clinical trial.

Keywords

D2C7-(scdsFv)-PE38KDEL Immunotoxin Convection-enhanced delivery Toxicity Rat 

Notes

Acknowledgments

We thank Terri Lucas and Lena Perdue for coordinating the study, Charles Pegram, David Soule, and Xiao-Guang Zhao for the formulation preparation, and Colleen Herbst, Meredith Weksler, and Fernando Orozco for their surgical assistance. We wish to thank Christopher Means and Theo Rhodes for their detailed assessment of the animals for the Functional Observation Battery (FOB) and assistance in preparing and analyzing the FOB data. We also thank Jenna Lewis for her editorial assistance. The study was funded by the following grant from the National Institutes of Health (NIH) of the United States: P01-CA154291-03 (to D.D. Bigner). This research was also supported in part by the Intramural Research Program of the NIH, National Cancer Institute, Center for Cancer Research.

Compliance with ethical standards

All applicable international, national, and/or institutional guidelines for the care and use of animals were followed.

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References

  1. 1.
    Ostrom QT, Gittleman H, Liao P, Rouse C, Chen Y, Dowling J, Wolinsky Y, Kruchko C, Barnholtz-Sloan J (2014) CBTRUS statistical report: primary brain and central nervous system tumors diagnosed in the United States in 2007–2011. Neuro Oncol 16(Suppl 4):iv1–iv63. doi: 10.1093/neuonc/nou223 CrossRefPubMedPubMedCentralGoogle Scholar
  2. 2.
    Wen PY, Kesari S (2008) Malignant gliomas in adults. N Engl J Med 359(5):492–507. doi: 10.1056/NEJMra0708126 CrossRefPubMedGoogle Scholar
  3. 3.
    Chandramohan V, Sampson JH, Pastan I, Bigner DD (2012) Toxin-based targeted therapy for malignant brain tumors. Clin Dev Immunol 2012:480429. doi: 10.1155/2012/480429 CrossRefPubMedPubMedCentralGoogle Scholar
  4. 4.
    Ahmad ZA, Yeap SK, Ali AM, Ho WY, Alitheen NB, Hamid M (2012) scFv antibody: principles and clinical application. Clin Dev Immunol 2012:980250. doi: 10.1155/2012/980250 CrossRefPubMedPubMedCentralGoogle Scholar
  5. 5.
    Shapira A, Benhar I (2010) Toxin-based therapeutic approaches. Toxins 2(11):2519–2583. doi: 10.3390/toxins2112519 CrossRefPubMedPubMedCentralGoogle Scholar
  6. 6.
    Pastan I, Hassan R, Fitzgerald DJ, Kreitman RJ (2006) Immunotoxin therapy of cancer. Nat Rev Cancer 6(7):559–565. doi: 10.1038/nrc1891 CrossRefPubMedGoogle Scholar
  7. 7.
    Piao H, Kuan CT, Chandramohan V, Keir ST, Pegram CN, Bao X, Mansson JE, Pastan IH, Bigner DD (2013) Affinity-matured recombinant immunotoxin targeting gangliosides 3′-isoLM1 and 3′,6′-isoLD1 on malignant gliomas. MAbs 5(5):748–762. doi: 10.4161/mabs.25860 CrossRefPubMedPubMedCentralGoogle Scholar
  8. 8.
    Salomon DS, Brandt R, Ciardiello F, Normanno N (1995) Epidermal growth factor-related peptides and their receptors in human malignancies. Crit Rev Oncol Hematol 19(3):183–232CrossRefPubMedGoogle Scholar
  9. 9.
    Chaffanet M, Chauvin C, Laine M, Berger F, Chedin M, Rost N, Nissou MF, Benabid AL (1992) EGF receptor amplification and expression in human brain tumours. Eur J Cancer 28(1):11–17CrossRefPubMedGoogle Scholar
  10. 10.
    Frederick L, Wang XY, Eley G, James CD (2000) Diversity and frequency of epidermal growth factor receptor mutations in human glioblastomas. Cancer Res 60(5):1383–1387PubMedGoogle Scholar
  11. 11.
    Chandramohan V, Bao X, Keir ST, Pegram CN, Szafranski SE, Piao H, Wikstrand CJ, McLendon RE, Kuan CT, Pastan IH, Bigner DD (2013) Construction of an immunotoxin, D2C7-(scdsFv)-PE38KDEL, targeting EGFRwt and EGFRvIII for brain tumor therapy. Clin Cancer Res 19(17):4717–4727. doi: 10.1158/1078-0432.CCR-12-3891 CrossRefPubMedPubMedCentralGoogle Scholar
  12. 12.
    Bao X, Chandramohan V, Keir ST, Pegram CN, McLendon RE, Kuan C-T, Pastan IH, Bigner DD (2013) Antitumor efficacy of D2C7-(scdsFv)-PE38KDEL, a novel immunotoxin targeting EGFRwt and EGFRvIII, by convection-enhanced delivery in orthotopic brain tumor mouse models. J Immunother Cancer 1(Suppl 1):P126. doi: 10.1186/2051-1426-1-s1-p126 CrossRefPubMedCentralGoogle Scholar
  13. 13.
    Ding D, Kanaly CW, Bigner DD, Cummings TJ, Herndon JE 2nd, Pastan I, Raghavan R, Sampson JH (2010) Convection-enhanced delivery of free gadolinium with the recombinant immunotoxin MR1-1. J Neuro-Oncol 98(1):1–7. doi: 10.1007/s11060-009-0046-7 CrossRefGoogle Scholar
  14. 14.
    Sampson JH, Brady M, Raghavan R, Mehta AI, Friedman AH, Reardon DA, Petry NA, Barboriak DP, Wong TZ, Zalutsky MR, Lally-Goss D, Bigner DD (2011) Colocalization of gadolinium-diethylene triamine pentaacetic acid with high-molecular-weight molecules after intracerebral convection-enhanced delivery in humans. Neurosurgery 69(3):668–676. doi: 10.1227/NEU.0b013e3182181ba8 CrossRefPubMedPubMedCentralGoogle Scholar
  15. 15.
    Moser VC (1991) Applications of a neurobehavioral screening battery. Int J Toxicol 10(6):661–669. doi: 10.3109/10915819109078658 CrossRefGoogle Scholar
  16. 16.
    Blanchette M, Fortin D (2011) Blood–brain barrier disruption in the treatment of brain tumors. Methods Mol Biol 686:447–463. doi: 10.1007/978-1-60761-938-3_23 CrossRefPubMedGoogle Scholar
  17. 17.
    Sampson JH, Akabani G, Archer GE, Berger MS, Coleman RE, Friedman AH, Friedman HS, Greer K, Herndon JE 2nd, Kunwar S, McLendon RE, Paolino A, Petry NA, Provenzale JM, Reardon DA, Wong TZ, Zalutsky MR, Pastan I, Bigner DD (2008) Intracerebral infusion of an EGFR-targeted toxin in recurrent malignant brain tumors. Neuro Oncol 10(3):320–329. doi: 10.1215/15228517-2008-012 CrossRefPubMedPubMedCentralGoogle Scholar
  18. 18.
    Mehta AI, Choi BD, Ajay D, Raghavan R, Brady M, Friedman AH, Pastan I, Bigner DD, Sampson JH (2012) Convection enhanced delivery of macromolecules for brain tumors. Curr Drug Discov Technol 9(4):305–310CrossRefPubMedGoogle Scholar
  19. 19.
    Sampson JH, Raghavan R, Brady M, Friedman AH, Bigner D (2011) Convection-enhanced delivery. J Neurosurg 115(3):463–464. doi: 10.3171/2010.11.JNS101801, discussion 465–466CrossRefPubMedGoogle Scholar
  20. 20.
    Kawamata T, Mori T, Sato S, Katayama Y (2007) Tissue hyperosmolality and brain edema in cerebral contusion. Neurosurg Focus 22(5):E5CrossRefPubMedGoogle Scholar
  21. 21.
    Bandaranayake NM, Nemoto EM, Stezoski SW (1978) Rat brain osmolality during barbiturate anesthesia and global brain ischemia. Stroke 9(3):249–254. doi: 10.1161/01.str.9.3.249 CrossRefPubMedGoogle Scholar
  22. 22.
    Noker PE, Fulton R, Pickett AC, Mann JF (2007) Multiple dose toxicity study of MR1-1-(MR1-1dsFvPE38KDEL, NSC-718877) in Sprague Dawley rats. Conducted at Southern Research Institute, Birmingham, Alabama and sponsored by National Cancer Institute, Bethesda, MarylandGoogle Scholar
  23. 23.
    Harling-Berg C, Knopf PM, Merriam J, Cserr HF (1989) Role of cervical lymph nodes in the systemic humoral immune response to human serum albumin microinfused into rat cerebrospinal fluid. J Neuroimmunol 25(2–3):185–193CrossRefPubMedGoogle Scholar
  24. 24.
    Palestro G, Mazzucco G, Navone R, Canese MG, Coda R, Novero D, Micca FB, Leonardo E (1980) Role of the T-cell system in glomerulonephritis induced in rats by human serum albumin (HSA). An immunological and morphological study. Virchows Arch B Cell Pathol Incl Mol Pathol 35(1):19–32CrossRefPubMedGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2016

Authors and Affiliations

  • Xuhui Bao
    • 1
  • Vidyalakshmi Chandramohan
    • 1
  • Randall P. Reynolds
    • 2
  • John N. Norton
    • 2
  • William C. Wetsel
    • 3
    • 4
    • 5
    • 6
  • Ramona M. Rodriguiz
    • 3
    • 4
  • Dipendra K. Aryal
    • 3
    • 4
  • Roger E. McLendon
    • 1
  • Edward D. Levin
    • 3
  • Neil A. Petry
    • 7
  • Michael R. Zalutsky
    • 1
    • 7
  • Bruce K. Burnett
    • 8
    • 9
  • Chien-Tsun Kuan
    • 1
  • Ira H. Pastan
    • 10
  • Darell D. Bigner
    • 1
    Email author
  1. 1.Preston Robert Tisch Brain Tumor Center at Duke and Department of PathologyDuke University Medical CenterDurhamUSA
  2. 2.Division of Laboratory Animal ResourcesDuke University Medical CenterDurhamUSA
  3. 3.Department of Psychiatry and Behavioral SciencesDuke University Medical CenterDurhamUSA
  4. 4.Mouse Behavioral and Neuroendocrine Analysis Core FacilityDuke University Medical CenterDurhamUSA
  5. 5.Department of NeurobiologyDuke University Medical CenterDurhamUSA
  6. 6.Department of Cell BiologyDuke University Medical CenterDurhamUSA
  7. 7.Department of RadiologyDuke University Medical CenterDurhamUSA
  8. 8.Duke Translational Medicine InstituteRegulatory Affairs OfficeDurhamUSA
  9. 9.School of MedicineDuke UniversityDurhamUSA
  10. 10.Laboratory of Molecular Biology, Center for Cancer ResearchNational Cancer Institute, National Institutes of HealthBethesdaUSA

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