HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS		QUICK SEARCH:   [advanced] Author: Keyword(s): Year:  Vol:  Page:

This Article Full Text Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Clark, R. Articles by Wells, J. A. Search for Related Content PubMed PubMed Citation Articles by Clark, R. Articles by Wells, J. A. Social Bookmarking                      What's this?

Volume 271, Number 36, Issue of September 6, 1996 pp. 21969-21977
©1996 by The American Society for Biochemistry and Molecular Biology, Inc.

---

Long-acting Growth Hormones Produced by Conjugation with Polyethylene Glycol

(Received for publication, April 5, 1996, and in revised form, June 6, 1996)

Ross Clark , Kenneth Olson ^§ , Germaine Fuh ^¶ , Melinda Marian , Deborah Mortensen , Glen Teshima ^§ , Suh Chang ^§ , Herman Chu ^'' , Venkat Mukku ^'' , Eleanor Canova-Davis ^§ , Todd Somers , Michael Cronin , Marjorie Winkler ^§ and James A. Wells ^¶

From the Departments of  Endocrine Research, ^§ Recovery Sciences, ^¶ Protein Engineering,  Pharmacokinetics, ^'' Quality Control, and  Bioorganic Chemistry, Genentech, Inc., South San Francisco, California 94080

Derivatives of human growth hormone (hGH) of increasing size were produced by reaction with the N-hydroxysuccinimide ester of polyethylene glycol-5000 (PEG₅₀₀₀), a 5-kDa reagent that selectively conjugates to primary amines. By adjusting the reaction conditions and purification procedure, it was possible to isolate hGH derivatives containing up to seven PEG moieties that altered the Stokes radius and thereby the effective molecular masses of the unmodified hormone from 22 to 300 kDa. Fortunately, the most reactive amines were ones that did not lie in either of the two sites important for receptor binding. Nonetheless, increasing the level of PEG modification linearly reduced the affinity of hGH for its receptor and increased the EC₅₀ in a cell-based assay up to 1500-fold. Most of the reduction in affinity was the result of slowing the association rate for the receptor. The clearance rate of hGH in rats was inversely proportional to effective molecular weight and closely fit a filtration model. We have tested the potency of these analogs by injecting them daily or every 6 days into hypophysectomized rats and determining the effects on body and organ growth. The efficacy of these analogs was optimal for hGH conjugated with 5 eq of PEG₅₀₀₀, and the potency was increased by about 10-fold compared with unmodified hGH. Such PEG-hGH derivatives show promise as long-acting alternatives to daily injections of hGH. More generally these studies show that improving hormone clearance properties, even at the expense of reducing receptor binding affinity, can lead to dramatic increases in hormone efficacy.

CiteULike    Complore    Connotea    Del.icio.us    Digg    Reddit    Technorati    What's this?

This article has been cited by other articles:

				M. Schlapschy, I. Theobald, H. Mack, M. Schottelius, H.-J. Wester, and A. Skerra Fusion of a recombinant antibody fragment with a homo-amino-acid polymer: effects on biophysical properties and prolonged plasma half-life Protein Eng. Des. Sel., June 26, 2007; (2007) gzm020v1. [Abstract] [Full Text] [PDF]

				G. N. Cox, M. S. Rosendahl, E. A. Chlipala, D. J. Smith, S. J. Carlson, and D. H. Doherty A Long-Acting, Mono-PEGylated Human Growth Hormone Analog Is a Potent Stimulator of Weight Gain and Bone Growth in Hypophysectomized Rats Endocrinology, April 1, 2007; 148(4): 1590 - 1597. [Abstract] [Full Text] [PDF]

				M.H. Parker, Y. Chen, F. Danehy, K. Dufu, J. Ekstrom, E. Getmanova, J. Gokemeijer, L. Xu, and D. Lipovsek Antibody mimics based on human fibronectin type three domain engineered for thermostability and high-affinity binding to vascular endothelial growth factor receptor two Protein Eng. Des. Sel., September 1, 2005; 18(9): 435 - 444. [Abstract] [Full Text] [PDF]

				M. Maamra, J. J. Kopchick, C. J. Strasburger, and R. J. M. Ross Pegvisomant, a Growth Hormone-Specific Antagonist, Undergoes Cellular Internalization J. Clin. Endocrinol. Metab., September 1, 2004; 89(9): 4532 - 4537. [Abstract] [Full Text] [PDF]

				P. Alvarez, C. A. Buscaglia, and O. Campetella Improving Protein Pharmacokinetics by Genetic Fusion to Simple Amino Acid Sequences J. Biol. Chem., January 30, 2004; 279(5): 3375 - 3381. [Abstract] [Full Text] [PDF]

				K. Yang, A. Basu, M. Wang, R. Chintala, M.-C. Hsieh, S. Liu, J. Hua, Z. Zhang, J. Zhou, M. Li, et al. Tailoring structure-function and pharmacokinetic properties of single-chain Fv proteins by site-specific PEGylation Protein Eng. Des. Sel., October 1, 2003; 16(10): 761 - 770. [Abstract] [Full Text] [PDF]

				S. Bernichtein, C. Kayser, K. Dillner, S. Moulin, J. J. Kopchick, J. A. Martial, G. Norstedt, O. Isaksson, P. A. Kelly, and V. Goffin Development of Pure Prolactin Receptor Antagonists J. Biol. Chem., September 19, 2003; 278(38): 35988 - 35999. [Abstract] [Full Text] [PDF]

				G. G. Kochendoerfer, S.-Y. Chen, F. Mao, S. Cressman, S. Traviglia, H. Shao, C. L. Hunter, D. W. Low, E. N. Cagle, M. Carnevali, et al. Design and Chemical Synthesis of a Homogeneous Polymer-Modified Erythropoiesis Protein Science, February 7, 2003; 299(5608): 884 - 887. [Abstract] [Full Text] [PDF]

				J. J. Kopchick, C. Parkinson, E. C. Stevens, and P. J. Trainer Growth Hormone Receptor Antagonists: Discovery, Development, and Use in Patients with Acromegaly Endocr. Rev., October 1, 2002; 23(5): 623 - 646. [Abstract] [Full Text] [PDF]

				M. S. Dennis, M. Zhang, Y. G. Meng, M. Kadkhodayan, D. Kirchhofer, D. Combs, and L. A. Damico Albumin Binding as a General Strategy for Improving the Pharmacokinetics of Proteins J. Biol. Chem., September 13, 2002; 277(38): 35035 - 35043. [Abstract] [Full Text] [PDF]

				S. K. Kelley, L. A. Harris, D. Xie, L. DeForge, K. Totpal, J. Bussiere, and J. A. Fox Preclinical Studies to Predict the Disposition of Apo2L/Tumor Necrosis Factor-Related Apoptosis-Inducing Ligand in Humans: Characterization of in Vivo Efficacy, Pharmacokinetics, and Safety J. Pharmacol. Exp. Ther., October 1, 2001; 299(1): 31 - 38. [Abstract] [Full Text] [PDF]

				R. B. Pepinsky, D. J. LePage, A. Gill, A. Chakraborty, S. Vaidyanathan, M. Green, D. P. Baker, E. Whalley, P. S. Hochman, and P. Martin Improved Pharmacokinetic Properties of a Polyethylene Glycol-Modified Form of Interferon-beta -1a with Preserved in Vitro Bioactivity J. Pharmacol. Exp. Ther., June 1, 2001; 297(3): 1059 - 1066. [Abstract] [Full Text]

				V. ROELFSEMA and R. G. CLARK The Growth Hormone and Insulin-Like Growth Factor Axis: Its Manipulation for the Benefit of Growth Disorders in Renal Failure J. Am. Soc. Nephrol., June 1, 2001; 12(6): 1297 - 1306. [Abstract] [Full Text]

				R. J. M. Ross, K. C. Leung, M. Maamra, W. Bennett, N. Doyle, M. J. Waters, and K. K. Y. Ho Binding and Functional Studies with the Growth Hormone Receptor Antagonist, B2036-PEG (Pegvisomant), Reveal Effects of Pegylation and Evidence That It Binds to a Receptor Dimer J. Clin. Endocrinol. Metab., April 1, 2001; 86(4): 1716 - 1723. [Abstract] [Full Text]

				P. J. Trainer, W. M. Drake, L. Katznelson, P. U. Freda, V. Herman-Bonert, A.J. van der Lely, E. V. Dimaraki, P. M. Stewart, K. E. Friend, M. L. Vance, et al. Treatment of Acromegaly with the Growth Hormone-Receptor Antagonist Pegvisomant N. Engl. J. Med., April 20, 2000; 342(16): 1171 - 1177. [Abstract] [Full Text] [PDF]

				Y. SEGEV, D. LANDAU, R. RASCH, A. FLYVBJERG, and M. PHILLIP Growth Hormone Receptor Antagonism Prevents Early Renal in Nonobese Diabetic Mice J. Am. Soc. Nephrol., November 1, 1999; 10(11): 2374 - 2381. [Abstract] [Full Text]

				L. Orsatti and F. M. Veronese An Unusual Coupling of Poly(Ethylene Glycol) to Tyrosine Residues in Epidermal Growth Factor Journal of Bioactive and Compatible Polymers, September 1, 1999; 14(5): 429 - 436. [Abstract] [PDF]

				J. Pato, M. Mora, I. Mezo, J. Seprodi, I. Teplan, B. Vincze, A. Kalnay, and I. Palyi Synthesis of New Poly(N-Vinyl Pyrrolidone-Co-Maleic Acid) Peptide Hormone Conjugates with Anticancer Activity Journal of Bioactive and Compatible Polymers, July 1, 1999; 14(4): 304 - 314. [Abstract] [PDF]

				M. De Serres, B. Ellis, J. E. Dillberger, S. K. Rudolph, J. T. Hutchins, C. M. Boytos, D. L. Weigl, and R. B. DePrince Immunogenicity of Thrombopoietin Mimetic Peptide GW395058 in BALB/c Mice and New Zealand White Rabbits: Evaluation of the Potential for Thrombopoietin Neutralizing Antibody Production in Man Stem Cells, July 1, 1999; 17(4): 203 - 209. [Abstract] [Full Text]

				M. O. Thorner, C. J. Strasburger, Z. Wu, M. Straume, M. Bidlingmaier, S. S. Pezzoli, K. Zib, J. C. Scarlett, and W. F. Bennett Growth Hormone (GH) Receptor Blockade with a PEG-Modified GH (B2036-PEG) Lowers Serum Insulin-Like Growth Factor-I but Does Not Acutely Stimulate Serum GH J. Clin. Endocrinol. Metab., June 1, 1999; 84(6): 2098 - 2103. [Abstract] [Full Text]

				Y.-N. Sun, H. J. Lee, R. R. Almon, and W. J. Jusko A Pharmacokinetic/Pharmacodynamic Model for Recombinant Human Growth Hormone Effects on Induction of Insulin-Like Growth Factor I in Monkeys J. Pharmacol. Exp. Ther., June 1, 1999; 289(3): 1523 - 1532. [Abstract] [Full Text]

				C. A. Buscaglia, J. Alfonso, O. Campetella, and A. C.C. Frasch Tandem Amino Acid Repeats From Trypanosoma cruzi Shed Antigens Increase the Half-Life of Proteins in Blood Blood, March 15, 1999; 93(6): 2025 - 2032. [Abstract] [Full Text] [PDF]

				D. Wu and W. M. Pardridge Neuroprotection with noninvasive neurotrophin delivery to the brain PNAS, January 5, 1999; 96(1): 254 - 259. [Abstract] [Full Text] [PDF]