Pharmaceutical significance of the cyclic imide form of recombinant human glial cell line derived neurotrophic factor.
OBJECTIVE
The purpose of this paper is to determine the significance of cyclic imide formation of an aspartic acid residue during storage on the pharmaceutical quality of a recombinant human glial cell line-derived neurotrophic factor (rhGDNF) formulation.


METHODS
A combination of chromatography, peptide mapping, mass spectroscopy, and protein sequencing was used to purify and characterize the degradation product. Circular dichroism, 1,8-ANS and heparin binding, melting temperature determination, bioassays, and preclinical pharmacokinetic and toxicology testing were performed to examine its equivalence to native rhGDNF.


RESULTS
The rhGDNF with cyclic imide at aspartic acid residue 96 showed identical activity, structure, pharmacokinetic profile, and toxicity profile to the native rhGDNF.


CONCLUSIONS
Formation of cyclic imide at aspartic acid residue 96 does not affect the pharmaceutical quality of the rhGDNF formulation.
Markell
D
D
Department of Pharmaceutics and Drug Delivery, Amgen, Inc., Thousand Oaks, CA 91320, USA.
Hui
J
J
Narhi
L
L
Lau
D
D
LeBel
C
C
Aparisio
D
D
Lile
J
J
Jing
S
S
Yui
D
D
Chang
B S
BS
D000818:Animals; D001224:Aspartic Acid / Q000737:chemistry; D002851:Chromatography, High Pressure Liquid; D002942:Circular Dichroism; D003500:Cyclization; D029721:Drosophila Proteins; D004355:Drug Stability; D004356:Drug Storage; D005728:Ganglia, Sympathetic / Q000166:cytology / Q000187:drug effects; D051100:Glial Cell Line-Derived Neurotrophic Factor; D051097:Glial Cell Line-Derived Neurotrophic Factor Receptors; D006493:Heparin / Q000378:metabolism; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007094:Imides / Q000737:chemistry*; D051379:Mice; D008934:Mitogens / Q000494:pharmacology; D009414:Nerve Growth Factors; D009419:Nerve Tissue Proteins / Q000737:chemistry* / Q000493:pharmacokinetics / Q000494:pharmacology*; D011518:Proto-Oncogene Proteins / Q000187:drug effects / Q000378:metabolism; D051096:Proto-Oncogene Proteins c-ret; D051381:Rats; D020794:Receptor Protein-Tyrosine Kinases / Q000187:drug effects / Q000378:metabolism; D011994:Recombinant Proteins / Q000737:chemistry / Q000494:pharmacology; D013378:Substantia Nigra / Q000166:cytology / Q000187:drug effects
D029721:Drosophila Proteins; C495039:GDNF protein, human; C495040:Gdnf protein, mouse; C495041:Gdnf protein, rat; D051100:Glial Cell Line-Derived Neurotrophic Factor; D051097:Glial Cell Line-Derived Neurotrophic Factor Receptors; D007094:Imides; D008934:Mitogens; D009414:Nerve Growth Factors; D009419:Nerve Tissue Proteins; D011518:Proto-Oncogene Proteins; D011994:Recombinant Proteins; D001224:Aspartic Acid; D006493:Heparin; D051096:Proto-Oncogene Proteins c-ret; D020794:Receptor Protein-Tyrosine Kinases; C416649:Ret protein, Drosophila