title: N-terminally PEGylated human interferon-beta-1a with improved pharmacokinetic properties and in vivo efficacy in a melanoma angiogenesis model.
abstract: PEGylation of IFN-alpha has been used successfully to improve the pharmacokinetic properties and efficacy of the drug. To prepare a PEGylated form of human interferon-beta-1a (IFN-beta-1a) suitable for testing in vivo, we have synthesized 20 kDa mPEG-O-2-methylpropionaldehyde and used it to modify the N-terminal alpha-amino group of the cytokine. The PEGylated protein retained approximately 50% of the activity of the unmodified protein and had significantly improved pharmacokinetic properties following intravenous administration in rats. The clearance and volume of distribution at steady state were reduced approximately 30-fold and approximately 4-fold, respectively, resulting in a significant increase in systemic exposure as determined by the area under the curve. The elimination half-life of the PEGylated protein was approximately 13-fold greater than for the unmodified protein. The unmodified and PEGylated proteins were tested for their ability to inhibit the formation of radially oriented blood vessels entering the periphery of human SK-MEL-1 melanoma tumors in athymic nude homozygous (nu/nu) mice. In a single dose comparison study, administration of 1 x 10(6) units of unmodified IFN-beta-1a resulted in a 29% reduction in vessel number, while 1 x 10(6) units of PEGylated IFN-beta-1a resulted in a 58% reduction. Both treatments resulted in statistically significant reductions in mean vessel number as compared to the vehicle (control)-treated mice, with the PEGylated IFN-beta-1a-treated mice showing a statistically significantly greater reduction in mean vessel number as compared to the unmodified IFN-beta-1a-treated mice. In a multiple versus single dose comparison study, daily administration of 1 x 10(6) units of unmodified IFN-beta-1a for 9 days resulted in a 51% reduction in vessel number, while a single dose of 1 x 10(6) units of the PEGylated protein resulted in a 66% reduction. Both treatments resulted in statistically significant reductions in mean vessel number as compared to the vehicle-treated mice, with the PEGylated IFN-beta-1a-treated mice showing a statistically significantly greater reduction in mean vessel number as compared to the unmodified IFN-beta-1a-treated mice. Therefore, the improved pharmacokinetic properties of the modified protein translated into improved efficacy. Since unmodified IFN-beta is used for the treatment of multiple sclerosis and hepatitis C virus infection, a PEGylated form of the protein such as 20 kDa mPEG-O-2-methylpropionaldehyde-modified IFN-beta-1a may serve as a useful adjunct for the treatment of these diseases. In addition, the antiangiogenic effects of PEGylated IFN-beta-1a may be harnessed for the treatment of certain cancers, either as a sole agent or in combination with other antitumor drugs.
authors.affiliation: BiogenIdec Inc., 14 Cambridge Center, Cambridge, Massachusetts 02142, USA. darren.baker@biogenidec.com
authors.forename: Darren P
authors.initials: DP
authors.lastname: Baker
authors.forename: Edward Y
authors.initials: EY
authors.lastname: Lin
authors.forename: Kochung
authors.initials: K
authors.lastname: Lin
authors.forename: Maria
authors.initials: M
authors.lastname: Pellegrini
authors.forename: Russell C
authors.initials: RC
authors.lastname: Petter
authors.forename: Ling Ling
authors.initials: LL
authors.lastname: Chen
authors.forename: Robert M
authors.initials: RM
authors.lastname: Arduini
authors.forename: Margot
authors.initials: M
authors.lastname: Brickelmaier
authors.forename: Dingyi
authors.initials: D
authors.lastname: Wen
authors.forename: Donna M
authors.initials: DM
authors.lastname: Hess
authors.forename: Liqing
authors.initials: L
authors.lastname: Chen
authors.forename: Donna
authors.initials: D
authors.lastname: Grant
authors.forename: Adrian
authors.initials: A
authors.lastname: Whitty
authors.forename: Alan
authors.initials: A
authors.lastname: Gill
authors.forename: Daniel J
authors.initials: DJ
authors.lastname: Lindner
authors.forename: R Blake
authors.initials: RB
authors.lastname: Pepinsky
mesh_terms: D000447:Aldehydes / Q000138:chemical synthesis / Q000493:pharmacokinetics / Q000627:therapeutic use*; D000818:Animals; D000998:Antiviral Agents / Q000737:chemistry / Q000493:pharmacokinetics / Q000627:therapeutic use*; D002460:Cell Line; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects; D004686:Encephalomyocarditis virus / Q000187:drug effects; D005260:Female; D006207:Half-Life; D006801:Humans; D000068556:Interferon beta-1a; D016899:Interferon-beta / Q000737:chemistry / Q000493:pharmacokinetics / Q000627:therapeutic use*; D008546:Melanoma, Experimental / Q000098:blood supply / Q000188:drug therapy* / Q000473:pathology; D008657:Metabolic Clearance Rate; D051379:Mice; D008819:Mice, Nude; D009368:Neoplasm Transplantation; D009389:Neovascularization, Pathologic / Q000517:prevention & control*; D011092:Polyethylene Glycols / Q000138:chemical synthesis / Q000493:pharmacokinetics / Q000627:therapeutic use*; D051381:Rats; D011917:Rats, Inbred Lew
chemical_list: D000447:Aldehydes; D000998:Antiviral Agents; C513579:mPEG-O-2-methylpropionaldehyde; D011092:Polyethylene Glycols; D016899:Interferon-beta; D000068556:Interferon beta-1a