title: Site-specific PEGylation enhances the pharmacokinetic properties and antitumor activity of interferon beta-1b.
abstract: Interferon beta (IFN-β) is widely used to ameliorate disease progression in patients with Multiple Sclerosis. IFN-β has a short half-life in humans, necessitating frequent administration for optimum effectiveness. Covalent modification of IFN-β with polyethylene glycol (PEG) improves the pharmacokinetic properties of the protein, but can adversely affect the protein's in vitro bioactivity. Random modification of lysine residues in IFN-β with amine-reactive PEGs decreased the in vitro bioactivity of the protein 50-fold, presumably due to modification of lysine residues near critical receptor binding sites. PEGylated IFN-β proteins that retained high in vitro bioactivity could be obtained by selective modification of the N-terminus of the protein with PEG. Here we use site-specific PEGylation technology (targeted attachment of a cysteine-reactive-PEG to an engineered cysteine residue in IFN-β) to identify several additional amino acid positions where PEG can be attached to IFN-β without appreciable loss of in vitro bioactivity. Unexpectedly, we found that most of the PEG-IFN-β analogs showed 11- to 78-fold improved in vitro bioactivities relative to their unPEGylated parent proteins and to IFN-β-1b. In vivo studies showed that a lead PEG-IFN-β protein had improved pharmacokinetic properties compared to IFN-β and was significantly more effective than IFN-β at inhibiting growth of a human tumor xenograft in athymic mice.
authors: 1 Bolder BioTechnology, Inc. , Boulder, Colorado.
authors: Ji I
authors: JI
authors: Lee
authors: Stephen P
authors: SP
authors: Eisenberg
authors: Mary S
authors: MS
authors: Rosendahl
authors: Elizabeth A
authors: EA
authors: Chlipala
authors: Jacquelyn D
authors: JD
authors: Brown
authors: Daniel H
authors: DH
authors: Doherty
authors: George N
authors: GN
authors: Cox
mesh_terms: D019943:Amino Acid Substitution; D000818:Animals; D000970:Antineoplastic Agents / Q000737:chemistry* / Q000493:pharmacokinetics / Q000494:pharmacology*; D049109:Cell Proliferation / Q000187:drug effects; D003545:Cysteine / Q000737:chemistry; D004305:Dose-Response Relationship, Drug; D006801:Humans; D000068576:Interferon beta-1b; D016899:Interferon-beta / Q000737:chemistry* / Q000493:pharmacokinetics / Q000494:pharmacology*; D008297:Male; D051379:Mice; D011092:Polyethylene Glycols / Q000737:chemistry*; D051381:Rats; D011994:Recombinant Proteins / Q000737:chemistry / Q000493:pharmacokinetics / Q000494:pharmacology; D023041:Xenograft Model Antitumor Assays
chemical_list: D000970:Antineoplastic Agents; D011994:Recombinant Proteins; D000068576:Interferon beta-1b; D011092:Polyethylene Glycols; D016899:Interferon-beta; D003545:Cysteine
references: EMBO J. 1992 Sep;11(9):3193-201
references: 1505514
references: Int J Clin Pract Suppl. 2002 Sep;(131):9-16
references: 12564807
references: Proc Natl Acad Sci U S A. 1987 Mar;84(6):1487-91
references: 3494243
references: J Interferon Cytokine Res. 1996 Oct;16(10):759-64
references: 8910759
references: Exp Hematol. 2006 Jun;34(6):697-704
references: 16728273
references: Proc Natl Acad Sci U S A. 1998 Nov 24;95(24):14411-6
references: 9826714
references: J Hepatol. 2002 Nov;37(5):655-9
references: 12399233
references: J Immunother Emphasis Tumor Immunol. 1993 Apr;13(3):191-200
references: 8471593
references: Bioconjug Chem. 2008 Jan;19(1):299-305
references: 18020402
references: J Interferon Res. 1992 Oct;12(5):329-36
references: 1431312
references: Methods Mol Biol. 1993;15:251-61
references: 21400283
references: J Interferon Cytokine Res. 1997 Nov;17(11):681-93
references: 9402106
references: DNA. 1986 Apr;5(2):137-48
references: 3519136
references: Cancer Biochem Biophys. 1984 Jun;7(2):175-86
references: 6467175
references: J Pharmacol Exp Ther. 2001 Jun;297(3):1059-66
references: 11356929
references: Bioconjug Chem. 2005 Jan-Feb;16(1):200-7
references: 15656592
references: CNS Drugs. 2011 Jan;25(1):67-88
references: 21128695
references: Biotechnology (N Y). 1990 Apr;8(4):343-6
references: 1366535
references: J Natl Cancer Inst. 1989 Jul 19;81(14):1061-8
references: 2472488
references: N Engl J Med. 2000 Dec 7;343(23):1666-72
references: 11106715
references: Curr Opin Struct Biol. 1995 Feb;5(1):114-21
references: 7773741
references: Clin Pharmacokinet. 2001;40(7):539-51
references: 11510630
references: Bioprocess Int. 2005 Apr;3(4):52-60
references: 29899681
references: J Clin Pharmacol. 2012 Jun;52(6):798-808
references: 21680782
references: Bioconjug Chem. 2005 Sep-Oct;16(5):1291-8
references: 16173810
references: Bioconjug Chem. 2006 Jan-Feb;17(1):179-88
references: 16417267
references: J Natl Cancer Inst. 1992 Aug 5;84(15):1185-90
references: 1378904
references: Proc Natl Acad Sci U S A. 1997 Oct 28;94(22):11813-8
references: 9342320
references: Pharm Res. 1996 Jul;13(7):996-1002
references: 8842035
references: Curr Opin Neurol. 2000 Jun;13(3):235-40
references: 10871245
references: J Neurol Sci. 2003 Feb 15;206(2):199-202
references: 12559511
references: Hepatology. 2007 Aug;46(2):315-23
references: 17654600
references: Neurology. 2001 Jun 12;56(11):1505-13
references: 11402107
references: Nature. 1981 Dec 10;294(5841):563-5
references: 6171735
references: Bioconjug Chem. 2001 Mar-Apr;12(2):195-202
references: 11312680
references: Endocrinology. 2007 Apr;148(4):1590-7
references: 17234711
references: J Pharmacol Exp Ther. 2011 Sep;338(3):984-96
references: 21690216
references: Bioconjug Chem. 2006 May-Jun;17(3):618-30
references: 16704199