Abstract
We have previously demonstrated that enterally administered heparin-binding EGF-like growth factor (HB-EGF) produced in Escherichia coli decreases the incidence and severity of intestinal injury in a neonatal rat model of necrotizing enterocolitis (NEC). In preparation for upcoming human clinical trials, large-scale production of HB-EGF according to Good Manufacturing Practice (GMP) has been successfully accomplished using a Pichia pastoris yeast system. The current studies used a neonatal rat model of NEC to elucidate several important preclinical characteristics of HB-EGF therapy. We found that enteral administration of HB-EGF (800 μg/kg/dose) four times a day effectively reduced the incidence and severity of NEC, that Pichia-derived HB-EGF was not significantly different from E. coli-derived HB-EGF in preventing NEC, that EGF was not superior to HB-EGF in preventing NEC, and that prophylactic administration of HB-EGF added to formula starting with the first feed or 12 h later significantly reduced the incidence of NEC, with no change in the incidence of NEC noted if HB-EGF was added to the formula starting 24, 48, or 72 h after birth. Thus, large-scale production of GMP-grade HB-EGF in Pichia pastoris yeast produces a biologically active molecule suitable for human clinical trials.
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Abbreviations
- EGFR:
-
epidermal growth factor receptor
- GMP:
-
good manufacturing process
- HB-EGF:
-
heparin-binding EGF-like growth factor
- NEC:
-
necrotizing enterocolitis
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Acknowledgements
The authors wish to thank Gregory Young, M.S., Ohio State University Center for Biostatistics, for his helpful comments and his assistance with the statistical analyses.
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Supported by Trillium Therapeutics, Inc. (Toronto, Ontario, Canada) (G.E.B., A.R.), NIH RO1 DK074611 (G.E.B.), NIHT32 RR-023260-02 (N.A.Z.), The Samuel J. Roessler Memorial Fellowship (N.A.Z.), and The Firefighters Endowment Fund of Nationwide Children's Hospital (X.Y.).
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Radulescu, A., Zorko, N., Yu, X. et al. Preclinical Neonatal Rat Studies of Heparin-Binding EGF-Like Growth Factor in Protection of the Intestines From Necrotizing Enterocolitis. Pediatr Res 65, 437–442 (2009). https://doi.org/10.1203/PDR.0b013e3181994fa0
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DOI: https://doi.org/10.1203/PDR.0b013e3181994fa0
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