Abstract
Fusion proteins made up of glucagon-like peptide 1 (GLP-1) and exendin-4 (EX-4) fused to a nonglycosylated form of human transferrin (GLP-1-Tf or EX-4-Tf) were produced and characterized. GLP-1-Tf activated the GLP-1 receptor, was resistant to inactivation by peptidases, and had a half-life of approximately 2 days, compared with 1 to 2 min for native GLP-1. GLP-1-Tf retained the acute, glucose-dependent insulin-secretory properties of native GLP-1 in diabetic animals and had a profound effect on proliferation of pancreatic β-cells. In addition, Tf and the fusion proteins did not cross the blood-brain-barrier but still reduced food intake after peripheral administration. EX-4-Tf proved to be as effective as EX-4 but had longer lived effects on blood glucose and food intake. This novel transferrin fusion technology could improve the pharmacology of various peptides.
Footnotes
This work was supported by the Intramural Research Program of the National Institutes of Health National Institute on Aging.
Article, publication date, and citation information can be found at http://jpet.aspetjournals.org.
doi:10.1124/jpet.110.166470.
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ABBREVIATIONS:
- GLP-1
- glucagon-like peptide 1
- DPP-IV
- dipeptidyl peptidase IV
- EX-4
- exendin-4
- Tf
- transferrin
- GLP-1-Tf
- glucagon-like peptide 1 fused to human transferrin
- EX-4-Tf
- exendin-4 fused to human transferrin
- PAGE
- polyacrylamide gel electrophoresis
- CHO
- Chinese hamster ovary
- ELISA
- enzyme-linked immunosorbent assay
- IPGTT
- intraperitoneal glucose tolerance test
- BrdU
- 5-bromo-2-deoxyuridine
- PBS
- phosphate-buffered saline
- HRP
- horseradish peroxidase
- h
- human
- CSF
- cerebrospinal fluid
- GLP-1R
- glucagon-like peptide 1 receptor
- AUC
- area under the curve
- AP
- area postrema
- NTS
- nuclei of the solitary tract
- PVH
- paraventricular nuclei of the hypothalamus
- LY2428757
- polyethylene glycol-GLP-1.
- Received January 27, 2010.
- Accepted May 21, 2010.
- U.S. Government work not protected by U.S. copyright
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