PMID- 21129326
OWN - NLM
STAT- MEDLINE
DCOM- 20110325
LR  - 20211020
IS  - 1932-2968 (Electronic)
IS  - 1932-2968 (Linking)
VI  - 4
IP  - 6
DP  - 2010 Nov 1
TI  - Optimization of the native glucagon sequence for medicinal purposes.
PG  - 1322-31
AB  - BACKGROUND: Glucagon is a life-saving medication used in the treatment of 
      hypoglycemia. It possesses poor solubility in aqueous buffers at or near 
      physiological pH values. At low and high pH, at which the peptide can be 
      formulated to concentrations of a milligram or more per milliliter, the chemical 
      integrity of the hormone is limited, as evidenced by the formation of multiple 
      degradation-related peptides. Consequently, the commercial preparation is 
      provided as a lyophilized solid with an acidic diluent and directions for 
      rendering it soluble at the time of use. Any unused material is recommended for 
      disposal immediately after initial use. METHODS: A set of glucagon analogs was 
      prepared by solid-phase peptide synthesis to explore the identification of a 
      glucagon analog with enhanced solubility and chemical stability at physiological 
      pH. The physical properties of the peptide analogs were studied by solubility 
      determination, high-performance chromatography, and mass spectral analysis. The 
      biochemical properties were determined in engineered human embryonic kidney cell 
      line 293 (HEK293) cells that overexpressed either the human glucagon or 
      glucagon-like peptide-1 (GLP-1) receptors linked to a luciferase reporter gene. 
      RESULTS: We observed the previously characterized formation of glucagon 
      degradation products upon incubation of the peptide in dilute acid for extended 
      periods or elevated temperature. Lowering the isoelectric point of the hormone 
      through the substitution of asparagine-28 with aspartic acid significantly 
      increased the solubility at physiological pH. Similarly, the C-terminal extension 
      (Cex) of the hormone with an exendin-based, 10-residue, C-terminal sequence 
      yielded a peptide of dramatically enhanced solubility. These two glucagon 
      analogs, D28 and Cex, maintained high potency and selectivity for the glucagon 
      receptor relative to GLP-1 receptor. CONCLUSIONS: Glucagon presents unique 
      structural challenges to the identification of an analog of high biological 
      activity and selectivity that also possesses sufficient aqueous solubility and 
      stability such that it might be developed as a ready-to-use medicine. The 
      glucagon analogs D28 and Cex demonstrated all of the chemical, physical, and 
      biochemical properties supportive of further study as potential clinical 
      candidates for treatment of hypoglycemia.
CI  - (c) 2010 Diabetes Technology Society.
FAU - Chabenne, Joseph R
AU  - Chabenne JR
AD  - Department of Chemistry, Indiana University, Bloomington, IN 47405-7102, USA.
FAU - DiMarchi, Maria A
AU  - DiMarchi MA
FAU - Gelfanov, Vasily M
AU  - Gelfanov VM
FAU - DiMarchi, Richard D
AU  - DiMarchi RD
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20101101
PL  - United States
TA  - J Diabetes Sci Technol
JT  - Journal of diabetes science and technology
JID - 101306166
RN  - 0 (GLP1R protein, human)
RN  - 0 (Glucagon-Like Peptide-1 Receptor)
RN  - 0 (Receptors, Glucagon)
RN  - 30KYC7MIAI (Aspartic Acid)
RN  - 7006-34-0 (Asparagine)
RN  - 9007-92-5 (Glucagon)
RN  - E0399OZS9N (Cyclic AMP)
SB  - IM
MH  - Amino Acid Sequence
MH  - Asparagine
MH  - Aspartic Acid
MH  - Cell Line
MH  - Chemistry, Pharmaceutical
MH  - Chromatography, High Pressure Liquid
MH  - Cyclic AMP/metabolism
MH  - Drug Stability
MH  - Genes, Reporter
MH  - Glucagon/analogs & derivatives/chemical synthesis/*chemistry/pharmacology
MH  - Glucagon-Like Peptide-1 Receptor
MH  - Humans
MH  - Hydrogen-Ion Concentration
MH  - Hypoglycemia/drug therapy
MH  - Isoelectric Point
MH  - Molecular Sequence Data
MH  - Receptors, Glucagon/drug effects/genetics/metabolism
MH  - Solubility
MH  - Spectrometry, Mass, Electrospray Ionization
MH  - Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
MH  - Temperature
MH  - Transfection
PMC - PMC3005041
EDAT- 2010/12/07 06:00
MHDA- 2011/03/26 06:00
PMCR- 2011/11/01
CRDT- 2010/12/07 06:00
PHST- 2010/12/07 06:00 [entrez]
PHST- 2010/12/07 06:00 [pubmed]
PHST- 2011/03/26 06:00 [medline]
PHST- 2011/11/01 00:00 [pmc-release]
AID - dst.4.6.1322 [pii]
AID - 10.1177/193229681000400605 [doi]
PST - epublish
SO  - J Diabetes Sci Technol. 2010 Nov 1;4(6):1322-31. doi: 10.1177/193229681000400605.