Highly glycosylated human alpha interferon: An insight into a new therapeutic candidate.
The type I human interferon alpha (hIFN-alpha) family consists of small proteins that exert a multiplicity of biological actions including antiviral, antiproliferative and immunomodulatory effects. However, though administration of recombinant hIFN-alpha2b is the current treatment for chronic hepatitis B and C and for some types of cancers, therapy outcomes have not been completely satisfactory. The short serum half-life and rapid clearance of the cytokine accounts for its low in vivo biological activity. Here we describe and characterize a long-acting rhIFN-alpha2b mutein, 4N-IFN, which has been created by introducing four N-glycosylation sites via site-directed mutagenesis. The hyperglycosylated protein was found to have a 25-fold longer plasma half-life than the non-glycosylated rhIFN-alpha2b, even greater than the commercial pegylated derivative Intron-A PEG. In addition, glycosylation increased the in vitro stability of the mutein against serum protease inactivation. Interestingly, despite its lower in vitro activity, 4N-IFN showed a markedly enhanced in vivo antitumor activity in human prostate carcinoma implanted in nude mice. MALDI-TOF MS and HPAEC-PAD carbohydrate analyses revealed the presence of high amounts of tetrasialylated (40%) and trisialylated (28%) N-glycan structures, which are consequently responsible for the improved characteristics of the cytokine, making 4N-IFN a new therapeutic candidate for viral and malignant diseases.
Ceaglio
Natalia
N
Laboratorio de Cultivos Celulares, Facultad de Bioquímica y Ciencias Biológicas, Universidad Nacional del Litoral, Ciudad Universitaria, Paraje El Pozo-C.C. 242 (S3000ZAA) Santa Fe, Argentina. nceaglio@fbcb.unl.edu.ar
Etcheverrigaray
Marina
M
Conradt
Harald S
HS
Grammel
Nicolas
N
Kratje
Ricardo
R
Oggero
Marcos
M
D000704:Analysis of Variance; D000818:Animals; D000970:Antineoplastic Agents / Q000737:chemistry* / Q000493:pharmacokinetics; D049109:Cell Proliferation / Q000187:drug effects; D002852:Chromatography, Ion Exchange; D004305:Dose-Response Relationship, Drug; D004355:Drug Stability; D005260:Female; D006031:Glycosylation; D006801:Humans; D000077190:Interferon alpha-2; D016898:Interferon-alpha / Q000737:chemistry* / Q000493:pharmacokinetics; D051379:Mice; D008819:Mice, Nude; D019158:N-Acetylneuraminic Acid / Q000032:analysis; D010447:Peptide Hydrolases; D011134:Polysaccharides / Q000032:analysis* / Q000737:chemistry; D051381:Rats; D017208:Rats, Wistar; D011994:Recombinant Proteins / Q000737:chemistry* / Q000493:pharmacokinetics; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D023041:Xenograft Model Antitumor Assays
D000970:Antineoplastic Agents; D000077190:Interferon alpha-2; D016898:Interferon-alpha; D011134:Polysaccharides; D011994:Recombinant Proteins; D010447:Peptide Hydrolases; D019158:N-Acetylneuraminic Acid