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Br J Cancer. 2001 April; 84(Suppl 1): 3–10.
PMCID: PMC2363897

Development and characterization of novel erythropoiesis stimulating protein (NESP)


Studies on human erythropoietin (EPO) demonstrated that there is a direct relationship between the sialic acid-containing carbohydrate content of the molecule and its serum half-life and in vivo biological activity, but an inverse relationship with its receptor-binding affinity. These observations led to the hypothesis that increasing the carbohydrate content, beyond that found naturally, would lead to a molecule with enhanced biological activity. Hyperglycosylated recombinant human EPO (rHuEPO) analogues were developed to test this hypothesis. Darbepoetin alfa (novel erythropoiesis stimulating protein, NESP, ARANESPTM, Amgen Inc, Thousand Oaks, CA), which was engineered to contain 5 N-linked carbohydrate chains (two more than rHuEPO), has been evaluated in preclinical animal studies. Due to its increased sialic acid-containing carbohydrate content, NESP is biochemically distinct from rHuEPO, having an increased molecular weight and greater negative charge. Compared with rHuEPO, it has an approximate 3-fold longer serum half-life, greater in vivo potency, and can be administered less frequently to obtain the same biological response. NESP is currently being evaluated in human clinical trials for treatment of anaemia and reduction in its incidence.© 2001 Cance Cance Cancer Research Campaign

Keywords: erythropoietin, darbepoetin alfa, pharmacokinetics, biological activity, carbohydrate, review

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Selected References

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  • Broudy VC, Lin N, Egrie J, de Haën C, Weiss T, Papayannopoulou T, Adamson JW. Identification of the receptor for erythropoietin on human and murine erythroleukemia cells and modulation by phorbol ester and dimethyl sulfoxide. Proc Natl Acad Sci U S A. 1988 Sep;85(17):6513–6517. [PubMed]
  • Browne JK, Cohen AM, Egrie JC, Lai PH, Lin FK, Strickland T, Watson E, Stebbing N. Erythropoietin: gene cloning, protein structure, and biological properties. Cold Spring Harb Symp Quant Biol. 1986;51(Pt 1):693–702. [PubMed]
  • Cazzola M, Mercuriali F, Brugnara C. Use of recombinant human erythropoietin outside the setting of uremia. Blood. 1997 Jun 15;89(12):4248–4267. [PubMed]
  • Cheetham JC, Smith DM, Aoki KH, Stevenson JL, Hoeffel TJ, Syed RS, Egrie J, Harvey TS. NMR structure of human erythropoietin and a comparison with its receptor bound conformation. Nat Struct Biol. 1998 Oct;5(10):861–866. [PubMed]
  • Cumming DA. Glycosylation of recombinant protein therapeutics: control and functional implications. Glycobiology. 1991 Mar;1(2):115–130. [PubMed]
  • D'Andrea AD, Lodish HF, Wong GG. Expression cloning of the murine erythropoietin receptor. Cell. 1989 Apr 21;57(2):277–285. [PubMed]
  • Davis JM, Arakawa T, Strickland TW, Yphantis DA. Characterization of recombinant human erythropoietin produced in Chinese hamster ovary cells. Biochemistry. 1987 May 5;26(9):2633–2638. [PubMed]
  • Delorme E, Lorenzini T, Giffin J, Martin F, Jacobsen F, Boone T, Elliott S. Role of glycosylation on the secretion and biological activity of erythropoietin. Biochemistry. 1992 Oct 20;31(41):9871–9876. [PubMed]
  • Dordal MS, Wang FF, Goldwasser E. The role of carbohydrate in erythropoietin action. Endocrinology. 1985 Jun;116(6):2293–2299. [PubMed]
  • Dubé S, Fisher JW, Powell JS. Glycosylation at specific sites of erythropoietin is essential for biosynthesis, secretion, and biological function. J Biol Chem. 1988 Nov 25;263(33):17516–17521. [PubMed]
  • Egrie JC, Strickland TW, Lane J, Aoki K, Cohen AM, Smalling R, Trail G, Lin FK, Browne JK, Hines DK. Characterization and biological effects of recombinant human erythropoietin. Immunobiology. 1986 Sep;172(3-5):213–224. [PubMed]
  • Eschbach JW, Egrie JC, Downing MR, Browne JK, Adamson JW. Correction of the anemia of end-stage renal disease with recombinant human erythropoietin. Results of a combined phase I and II clinical trial. N Engl J Med. 1987 Jan 8;316(2):73–78. [PubMed]
  • Eschbach JW, Abdulhadi MH, Browne JK, Delano BG, Downing MR, Egrie JC, Evans RW, Friedman EA, Graber SE, Haley NR, et al. Recombinant human erythropoietin in anemic patients with end-stage renal disease. Results of a phase III multicenter clinical trial. Ann Intern Med. 1989 Dec 15;111(12):992–1000. [PubMed]
  • Evans RW, Rader B, Manninen DL. The quality of life of hemodialysis recipients treated with recombinant human erythropoietin. Cooperative Multicenter EPO Clinical Trial Group. JAMA. 1990 Feb 9;263(6):825–830. [PubMed]
  • Fukuda MN, Sasaki H, Lopez L, Fukuda M. Survival of recombinant erythropoietin in the circulation: the role of carbohydrates. Blood. 1989 Jan;73(1):84–89. [PubMed]
  • Goldwasser E, Kung CK, Eliason J. On the mechanism of erythropoietin-induced differentiation. 13. The role of sialic acid in erythropoietin action. J Biol Chem. 1974 Jul 10;249(13):4202–4206. [PubMed]
  • Higuchi M, Oh-eda M, Kuboniwa H, Tomonoh K, Shimonaka Y, Ochi N. Role of sugar chains in the expression of the biological activity of human erythropoietin. J Biol Chem. 1992 Apr 15;267(11):7703–7709. [PubMed]
  • Krantz SB. Erythropoietin. Blood. 1991 Feb 1;77(3):419–434. [PubMed]
  • Lacombe C, Mayeux P. Biology of erythropoietin. Haematologica. 1998 Aug;83(8):724–732. [PubMed]
  • Lai PH, Everett R, Wang FF, Arakawa T, Goldwasser E. Structural characterization of human erythropoietin. J Biol Chem. 1986 Mar 5;261(7):3116–3121. [PubMed]
  • Lodish HF, Hilton DJ, Klingmüller U, Watowich SS, Wu H. The erythropoietin receptor: biogenesis, dimerization, and intracellular signal transduction. Cold Spring Harb Symp Quant Biol. 1995;60:93–104. [PubMed]
  • LOWY PH, KEIGHLEY G, BORSOOK H. Inactivation of erythropoietin by neuraminidase and by mild substitution reactions. Nature. 1960 Jan 9;185:102–103. [PubMed]
  • Lukowsky WA, Painter RH. Studies on the role of sialic acid in the physical and biological properties of erythropoietin. Can J Biochem. 1972 Aug;50(8):909–917. [PubMed]
  • Macdougall IC, Gray SJ, Elston O, Breen C, Jenkins B, Browne J, Egrie J. Pharmacokinetics of novel erythropoiesis stimulating protein compared with epoetin alfa in dialysis patients. J Am Soc Nephrol. 1999 Nov;10(11):2392–2395. [PubMed]
  • Markham A, Bryson HM. Epoetin alfa. A review of its pharmacodynamic and pharmacokinetic properties and therapeutic use in nonrenal applications. Drugs. 1995 Feb;49(2):232–254. [PubMed]
  • Miyake T, Kung CK, Goldwasser E. Purification of human erythropoietin. J Biol Chem. 1977 Aug 10;252(15):5558–5564. [PubMed]
  • Narhi LO, Arakawa T, Aoki KH, Elmore R, Rohde MF, Boone T, Strickland TW. The effect of carbohydrate on the structure and stability of erythropoietin. J Biol Chem. 1991 Dec 5;266(34):23022–23026. [PubMed]
  • Recny MA, Scoble HA, Kim Y. Structural characterization of natural human urinary and recombinant DNA-derived erythropoietin. Identification of des-arginine 166 erythropoietin. J Biol Chem. 1987 Dec 15;262(35):17156–17163. [PubMed]
  • Sasaki H, Bothner B, Dell A, Fukuda M. Carbohydrate structure of erythropoietin expressed in Chinese hamster ovary cells by a human erythropoietin cDNA. J Biol Chem. 1987 Sep 5;262(25):12059–12076. [PubMed]
  • Sasaki H, Ochi N, Dell A, Fukuda M. Site-specific glycosylation of human recombinant erythropoietin: analysis of glycopeptides or peptides at each glycosylation site by fast atom bombardment mass spectrometry. Biochemistry. 1988 Nov 15;27(23):8618–8626. [PubMed]
  • Skehel JJ, Stevens DJ, Daniels RS, Douglas AR, Knossow M, Wilson IA, Wiley DC. A carbohydrate side chain on hemagglutinins of Hong Kong influenza viruses inhibits recognition by a monoclonal antibody. Proc Natl Acad Sci U S A. 1984 Mar;81(6):1779–1783. [PubMed]
  • Sowade B, Sowade O, Möcks J, Franke W, Warnke H. The safety of treatment with recombinant human erythropoietin in clinical use: a review of controlled studies. Int J Mol Med. 1998 Feb;1(2):303–314. [PubMed]
  • Spivak JL, Hogans BB. The in vivo metabolism of recombinant human erythropoietin in the rat. Blood. 1989 Jan;73(1):90–99. [PubMed]
  • Syed RS, Reid SW, Li C, Cheetham JC, Aoki KH, Liu B, Zhan H, Osslund TD, Chirino AJ, Zhang J, et al. Efficiency of signalling through cytokine receptors depends critically on receptor orientation. Nature. 1998 Oct 1;395(6701):511–516. [PubMed]
  • Takeuchi M, Takasaki S, Miyazaki H, Kato T, Hoshi S, Kochibe N, Kobata A. Comparative study of the asparagine-linked sugar chains of human erythropoietins purified from urine and the culture medium of recombinant Chinese hamster ovary cells. J Biol Chem. 1988 Mar 15;263(8):3657–3663. [PubMed]
  • Tsuda E, Goto M, Murakami A, Akai K, Ueda M, Kawanishi G, Takahashi N, Sasaki R, Chiba H, Ishihara H, et al. Comparative structural study of N-linked oligosaccharides of urinary and recombinant erythropoietins. Biochemistry. 1988 Jul 26;27(15):5646–5654. [PubMed]
  • Winearls CG, Oliver DO, Pippard MJ, Reid C, Downing MR, Cotes PM. Effect of human erythropoietin derived from recombinant DNA on the anaemia of patients maintained by chronic haemodialysis. Lancet. 1986 Nov 22;2(8517):1175–1178. [PubMed]

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