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 microg/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.
Publication types
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Comparative Study
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Research Support, N.I.H., Extramural
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Research Support, Non-U.S. Gov't
MeSH terms
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Animals
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Animals, Newborn
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Disease Models, Animal
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Dose-Response Relationship, Drug
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Drug Administration Schedule
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Enteral Nutrition
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Enterocolitis, Necrotizing / etiology
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Enterocolitis, Necrotizing / pathology
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Enterocolitis, Necrotizing / prevention & control*
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Epidermal Growth Factor / pharmacology
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Escherichia coli / genetics
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Escherichia coli / metabolism
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Gastrointestinal Agents / administration & dosage
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Gastrointestinal Agents / pharmacology*
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Heparin-binding EGF-like Growth Factor
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Humans
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Hypoxia / complications
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Intercellular Signaling Peptides and Proteins / administration & dosage
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Intercellular Signaling Peptides and Proteins / biosynthesis
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Intercellular Signaling Peptides and Proteins / genetics
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Intercellular Signaling Peptides and Proteins / pharmacology*
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Intestines / drug effects*
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Intestines / pathology
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Lipopolysaccharides
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Pichia / genetics
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Pichia / metabolism
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Rats
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Rats, Sprague-Dawley
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Recombinant Proteins / pharmacology
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Severity of Illness Index
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Time Factors
Substances
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Gastrointestinal Agents
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HBEGF protein, human
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Hbegf protein, rat
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Heparin-binding EGF-like Growth Factor
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Intercellular Signaling Peptides and Proteins
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Lipopolysaccharides
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Recombinant Proteins
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Epidermal Growth Factor