Article thumbnail

A Better Anti-Diabetic Recombinant Human Fibroblast Growth Factor 21 (rhFGF21) Modified with Polyethylene Glycol

By Zhifeng Huang, Huiyan Wang, Meifei Lu, Chuanchuan Sun, Xiaoping Wu, Yi Tan, Chaohui Ye, Guanghui Zhu, Xiaojie Wang, Lu Cai and Xiaokun Li

Abstract

As one of fibroblast growth factor (FGF) family members, FGF21 has been extensively investigated for its potential as a drug candidate to combat metabolic diseases. In the present study, recombinant human FGF21 (rhFGF21) was modified with polyethylene glycol (PEGylation) in order to increase its in vivo biostabilities and therapeutic potency. At N-terminal residue rhFGF21 was site-selectively PEGylated with mPEG20 kDa-butyraldehyde. The PEGylated rhFGF21 was purified to near homogeneity by Q Sepharose anion-exchange chromatography. The general structural and biochemical features as well as anti-diabetic effects of PEGylated rhFGF21 in a type 2 diabetic rat model were evaluated. By N-terminal sequencing and MALDI-TOF mass spectrometry, we confirmed that PEG molecule was conjugated only to the N-terminus of rhFGF21. The mono-PEGylated rhFGF21 retained the secondary structure, consistent with the native rhFGF21, but its biostabilities, including the resistance to physiological temperature and trypsinization, were significantly enhanced. The in vivo immunogenicity of PEGylated rhFGF21 was significantly decreased, and in vivo half-life time was significantly elongated. Compared to the native form, the PEGylated rhFGF21 had a similar capacity of stimulating glucose uptake in 3T3-L1 cells in vitro, but afforded a significantly long effect on reducing blood glucose and triglyceride levels in the type 2 diabetic animals. These results suggest that the PEGylated rhFGF21 is a better and more effective anti-diabetic drug candidate than the native rhFGF21 currently available. Therefore, the PEGylated rhFGF21 may be potentially applied in clinics to improve the metabolic syndrome for type 2 diabetic patients

Topics: Research Article
Publisher: Public Library of Science
OAI identifier: oai:pubmedcentral.nih.gov:3108960
Provided by: PubMed Central

To submit an update or takedown request for this paper, please submit an Update/Correction/Removal Request.

Suggested articles

Citations

  1. (1976). A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding.
  2. (2009). Acute glucose-lowering and insulin-sensitizing action of FGF21 in insulin resistant mouse models----Association with liver and adipose tissue effects.
  3. (1973). Automated Edman degradation: the protein sequenator.
  4. (2008). betaKlotho is required for fibroblast growth factor (FGF) 21 signaling through FGF receptor (FGFR) 1c and FGFR3c.
  5. (2007). BetaKlotho is required for metabolic activity of fibroblast growth factor 21.
  6. (2004). Beyond the red cell: pegylation of other blood cells and tissues.
  7. (1987). Chemical modification of recombinant interleukin 2 by polyethylene glycol increases its potency in the murine Meth A sarcoma model.
  8. (1992). Detection and molecular weight determination of polyethylene glycol-modified hirudin by staining after sodium dodecyl sulfatepolyacrylamide gel electrophoresis.
  9. (2009). Different roles of N- and C- termini in the functional activity of FGF21.
  10. (2005). FGF-21 as a novel metabolic regulator.
  11. (2010). FGF-21 enhances islet engraftment in mouse syngeneic islet transplantation model.
  12. (2008). FGF21 is an Akt-regulated myokine.
  13. (2009). FGF21 N- and Ctermini play different roles in receptor interaction and activation.
  14. (2008). FGF21/FGF-21 receptor interaction and activation is determined by betaKlotho.
  15. (2004). Fibroblast growth factor 19 increases metabolic rate and reverses dietary and leptindeficient diabetes.
  16. (2008). Fibroblast growth factor 21 corrects obesity in mice.
  17. (2009). Fibroblast growth factor 21 reverses hepatic steatosis, increases energy expenditure, and improves insulin sensitivity in diet-induced obese mice.
  18. (2006). Fibroblast growth factor-21 improves pancreatic beta-cell function and survival by activation of extracellular signal-regulated kinase 1/2 and Akt signaling pathways.
  19. (2010). High-level expression and purification of soluble recombinant FGF21 protein by SUMO fusion in Escherichia coli.
  20. (2000). Identification of a novel FGF, FGF-21, preferentially expressedinthe liver.
  21. (2008). Increasing solubility of proteins and peptides by site-specific modification with betaine.
  22. (2008). Mero A
  23. (2007). Molecular insights into the klotho-dependent, endocrine mode of action of fibroblast growth factor 19 subfamily members.
  24. (2002). Mono-N-terminal poly(ethylene glycol)-protein conjugates.
  25. (2009). PEG conjugates in clinical development or use as anticancer agents: an overview.
  26. (2008). Physiological regulation and disorders of phosphate metabolism--pivotal role of fibroblast growth factor 23.
  27. (2010). Poly(ethylene glycol) modification enhances penetration of fibroblast growth factor 2 to injured spinal cord tissue from an intrathecal delivery system.
  28. (1990). Polyethylene glycol modification of the monoclonal antibody A7 enhances its tumor localization.
  29. (1994). Pseudomonas exotoxin A mutants. Replacement of surface exposed residues in domain II with cysteine residues that Modified rhFGF21 and Better Anti-Diabetic Effect PLoS
  30. (2009). Role of FGF19 induced FGFR4 activation in the regulation of glucose homeostasis. Aging
  31. (2008). Serum FGF21 levels are increased in obesity and are independently associated with the metabolic syndrome in humans.
  32. (2007). Site-specific pegylation of G-CSF by reversible denaturation.
  33. (2006). Sitespecific attachment of polyethylene glycol-like oligomers to proteins and peptides.
  34. (2007). Solid-phase PEGylation of recombinant interferon alpha-2a for site-specific modification: process performance, characterization, and in vitro bioactivity.
  35. (2007). The emerging role of the fibroblast growth factor-23-klotho axis in renal regulation of phosphate homeostasis.
  36. (2009). The FGF family: biology, pathophysiology and therapy.
  37. (1991). The fibroblast growth factor family.
  38. (1998). The heparan sulfate-fibroblast growth factor family: diversity of structure and function.
  39. (2007). The metabolic state of diabetic monkeys is regulated by fibroblast growth factor-21.
  40. (2007). Tissue-specific expression of betaKlotho and fibroblast growth factor (FGF) receptor isoforms determines metabolic activity of FGF19 and FGF21.