title: Conjugation of PolyPEG to interferon alpha extends serum half-life while maintaining low viscosity of the conjugate.
abstract: The covalent attachment of poly(ethylene glycol) (PEG) to therapeutic proteins is a commonly used approach for extending in vivo half-lives. A potential limitation of this PEGylation strategy is the adverse effect of PEG on conjugate viscosity. Interferon-alpha (IFN) was conjugated via its N-terminal amino group by reductive amination to α-aldehyde functional comb-shaped PolyPEG polymers (50 and 70 kDa) and to linear PEG (30 kDa). In vitro potencies of the purified PEGylated IFN conjugates were measured by reporter gene assay using a HEK293P/ISRE-SEAP cell line. IFN levels were measured in rats following intravenous injection. Viscosities of various linear PEG and PolyPEG polymers along with the polymer-IFN conjugates were determined using a rotational rheometer with cone-and-plate geometry. In vitro potencies and half-lives of the PEGylated IFN conjugates were compared with those of the marketed branched PEG-IFN conjugate PEGASYS. Both PolyPEG-IFN conjugates retained a similar potency as that of the marketed comparator, whereas the linear PEG-IFN conjugate potency was greater. All conjugates showed extended half-lives compared to that of naked IFN, with the PolyPEG conjugates exhibiting the longest half-lives and the linear PEG conjugate, the shortest. Viscosity analysis showed that the linear PEG-IFN conjugate was over twice as viscous as both PolyPEG conjugates. Taken together, this work demonstrates the potential of PolyPEG conjugation to therapeutic proteins as a novel tool for optimizing pharmacokinetic profiles in a way that potentially allows administration of high-dose formulations because of lower conjugate viscosity.
authors: †Sandoz Biopharmaceuticals, Mengeš, Lek Pharmaceuticals d.d., Kolodvorska 27, SI-1234 Mengeš, Slovenia.
authors: B
authors: B
authors: Podobnik
authors: ‡Novartis Pharma AG, CH-4056 Basel, Switzerland.
authors: B
authors: B
authors: Helk
authors: †Sandoz Biopharmaceuticals, Mengeš, Lek Pharmaceuticals d.d., Kolodvorska 27, SI-1234 Mengeš, Slovenia.
authors: V
authors: V
authors: Smilović
authors: †Sandoz Biopharmaceuticals, Mengeš, Lek Pharmaceuticals d.d., Kolodvorska 27, SI-1234 Mengeš, Slovenia.
authors: Š
authors: Š
authors: Škrajnar
authors: †Sandoz Biopharmaceuticals, Mengeš, Lek Pharmaceuticals d.d., Kolodvorska 27, SI-1234 Mengeš, Slovenia.
authors: K
authors: K
authors: Fidler
authors: †Sandoz Biopharmaceuticals, Mengeš, Lek Pharmaceuticals d.d., Kolodvorska 27, SI-1234 Mengeš, Slovenia.
authors: S
authors: S
authors: Jevševar
authors: §PolyTherics Ltd, Babraham Research Campus, Babraham, Cambridge CB22 3AT, United Kingdom.
authors: A
authors: A
authors: Godwin
authors: ∥PolyTherics Ltd, Unit 4, Vanguard Centre, University of Warwick Science Park, Coventry, CV4 7EZ, United Kingdom.
authors: P
authors: P
authors: Williams
mesh_terms: D000818:Animals; D005260:Female; D057809:HEK293 Cells; D006207:Half-Life; D006801:Humans; D016898:Interferon-alpha / Q000097:blood* / Q000737:chemistry*; D011092:Polyethylene Glycols / Q000737:chemistry* / Q000378:metabolism*; D051381:Rats; D017208:Rats, Wistar; D014783:Viscosity
chemical_list: D016898:Interferon-alpha; D011092:Polyethylene Glycols