A new method to produce monoPEGylated dimeric cytokines shown with human interferon-α2b.
We have adapted the dock-and-lock (DNL) method into a novel PEGylation technology using human interferon-α2b (IFN-α2b) as an example. Central to DNL is a pair of distinct protein domains involved in the natural association between cAMP-dependent protein kinase (PKA) and A-kinase anchoring proteins (AKAPs). These domains serve as linkers for site-specific conjugation of poly(ethylene glycol) (PEG) to a dimeric form of IFN-α2b. The combination of a fusion protein comprising IFN-α2b and the dimerization-and-docking domain (DDD) of PKA with a PEG-derivatized anchoring domain (AD) of an interactive AKAP results in facile formation of a trimeric complex containing two copies of IFN-α2b and a single site-specifically linked PEG chain. Three such monoPEGylated dimers of IFN-α2b have been generated, the first with a 20 kDa linear PEG, referred to as α2b-362, the second with a 30 kDa linear PEG (α2b-413), and the third with a 40 kDa branched PEG (α2b-457). All three retained antiviral and antitumor activity in vitro and showed improved pharmacokinetic properties in mice, which translated into potent and prolonged therapeutic efficacy in the Daudi human lymphoma xenograft model. We anticipate wide applicability of the DNL method for developing long-acting therapeutics that are dimeric and monoPEGylated with the increased bioavailability allowing for less frequent dosing.
Chang
Chien-Hsing
CH
IBC Pharmaceuticals Inc., Morris Plains, New Jersey 07950, USA.
Rossi
Edmund A
EA
Cardillo
Thomas M
TM
Nordstrom
Diane L
DL
McBride
William J
WJ
Goldenberg
David M
DM
D054758:A Kinase Anchor Proteins / Q000737:chemistry*; D000595:Amino Acid Sequence; D000818:Animals; D000970:Antineoplastic Agents / Q000138:chemical synthesis* / Q000378:metabolism* / Q000493:pharmacokinetics*; D000998:Antiviral Agents / Q000138:chemical synthesis* / Q000378:metabolism*; D017868:Cyclic AMP-Dependent Protein Kinases / Q000737:chemistry*; D003692:Delayed-Action Preparations / Q000138:chemical synthesis* / Q000378:metabolism* / Q000493:pharmacokinetics*; D019281:Dimerization; D015195:Drug Design; D005260:Female; D006801:Humans; D000077190:Interferon alpha-2; D016898:Interferon-alpha / Q000737:chemistry / Q000378:metabolism / Q000493:pharmacokinetics; D008223:Lymphoma / Q000188:drug therapy*; D051379:Mice; D016513:Mice, SCID; D008969:Molecular Sequence Data; D011092:Polyethylene Glycols / Q000737:chemistry*; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D011994:Recombinant Proteins; D023041:Xenograft Model Antitumor Assays
D054758:A Kinase Anchor Proteins; D000970:Antineoplastic Agents; D000998:Antiviral Agents; D003692:Delayed-Action Preparations; D000077190:Interferon alpha-2; D016898:Interferon-alpha; D011993:Recombinant Fusion Proteins; D011994:Recombinant Proteins; D011092:Polyethylene Glycols; D017868:Cyclic AMP-Dependent Protein Kinases