Positional Isomers of Monopegylated Interferon α-2a: Isolation, Characterization, and Biological Activity

Monkarsh, Seth P.; Ma, Yizhou; Aglione, Anthony; Bailon, Pascal; Ciolek, Doreen; Debarbieri, Barabara; Graves, Mary C.; Hollfelder, Kurt; Michel, Hanspeter; Palleroni, Alicia V.; Porter, Jill E.; Russoman, Emil; Roy, Swapan Kumar; Pan, Yu‐Ching E.

doi:10.1006/abio.1997.2128

Cited by 109 publications

(70 citation statements)

References 12 publications

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“…However, one exception is the inability to resolve by HPIEX the neighboring peaks of 5-kDa Lys 131 and 5-kDa Lys 164 , hence the lower purity estimation by analytical HPIEX relative to HPSEC. The HPIEX chromatography profiles and positional isomer elution order in this study matched those reported previously (12,15,19). In addition, the identity and purity of the purified positional isomers were confirmed by peptide mapping analysis.…”

Section: Confirmation Of Differential Activity Between 12-kda Monopegsupporting

confidence: 85%

Site of Pegylation and Polyethylene Glycol Molecule Size Attenuate Interferon-α Antiviral and Antiproliferative Activities through the JAK/STAT Signaling Pathway

Grace

Lee²,

Bradshaw

et al. 2005

Journal of Biological Chemistry

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Therapeutic pegylated interferon-␣s (IFN-␣) are mixtures of positional isomers that have been monopegylated at specific sites on the core IFN-␣ molecule. The pegylation results in lower in vitro specific activity associated with the core IFN-␣ molecule that is related to the site of pegylation and size of polyethylene glycol (PEG) attached. We prepared purified, homogeneous, positional pegylation isomers of IFN-␣2b that were monopegylated using 5-30-kDa linear PEG molecules attached at 7 primary reactive amino acid residues: Chronic hepatitis C is considered one of the major causes of chronic liver disease, cirrhosis, and hepatocellular carcinoma and is the most common reason for liver transplantation in the United States (1). It is estimated that there are 3 million chronically infected individuals in the United States and over 170 million worldwide (1). Treatment of hepatitis C has evolved from the use of interferon-␣ (IFN-␣), 1 either alone or in combination with ribavirin, to the newer pegylated interferons (PEGIFNs), which have provided a dramatic increase in virological response, especially in combination with ribavirin. Standard IFN-␣ therapy has a short (Ͻ12-h) half-life that requires subcutaneous injection three times weekly to maintain effective levels in the blood (2). The short half-life of IFN-␣ has led to the development of longer lasting preparations achieved by the attachment of a large polyethylene glycol (PEG) molecule directly to IFN-␣. Two different commercial preparations of PEG-IFN-␣ have been developed for clinical use, PEG-IFN-␣2b (PEG-INTRON®) and PEG-IFN-␣2a (Pegasys®); both have long half-lives (40 and 80 h, respectively) that permit once weekly administration (3). Both of these preparations have been demonstrated to be effective for the treatment of patients with hepatitis C (4), and clinical trial results have shown further that both of the pegylated molecules produce sustained viral response rates superior to those achieved with their respective standard IFN-␣s (5-7).Whereas pegylation has proven to be highly effective for slowing the clearance of biological molecules, including IFN-␣, and thus increasing serum half-life, it has been shown to also modify in vitro biological activity (8). For instance, we have reported that pegylation of IFN-␣2b with a 12-kDa linear PEG molecule results in a preparation that has a specific activity of 28% relative to IFN-␣2b; the loss in activity was not due to structural perturbation of the core IFN-␣2b core protein (9). Other groups have reported that pegylation of IFN-␣2a with a 40-kDa branched PEG molecule results in a preparation that contains from 1 to 7% relative specific activity compared with IFN-␣2a (10, 11). These two pegylated interferon-␣s (PEG-IFN␣s) differ substantially in their postpegylation constituent properties. PEG-IFN-␣2b has a 12-kDa linear PEG molecule attached using succinimidyl carbonate polyethylene glycol (SC-PEG) chemistry via a covalent urethane-like bond to the IFN␣2b protein (12). The pegylation linkage process results...

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Section: Confirmation Of Differential Activity Between 12-kda Monopegsupporting

confidence: 85%

Site of Pegylation and Polyethylene Glycol Molecule Size Attenuate Interferon-α Antiviral and Antiproliferative Activities through the JAK/STAT Signaling Pathway

Grace

Lee²,

Bradshaw

et al. 2005

Journal of Biological Chemistry

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“…Several approaches have been attempted to increase peptide half life and stability [2,3]. Many of the suggested strategies include a chemical modification of the peptide itself by means of, e.g., pegylation [4,5] or glycosylation [6]. However, altering the chemical structure of the peptide drug risks impairing its therapeutic effect.…”

Section: Introductionmentioning

confidence: 99%

PEG-stabilized lipid disks as carriers for amphiphilic antimicrobial peptides

Zetterberg

Reijmar

Pränting

et al. 2011

Journal of Controlled Release

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This is an accepted version of a paper published in Journal of Controlled Release. This paper has been peer-reviewed but does not include the final publisher proof-corrections or journal pagination.Citation for the published paper: Zetterberg, M., Reijmar, K., Pränting, M., Engström, Å., Andersson, D. et al. (2011) AbstractAntimicrobial peptides hold potential as a possible alternative, or complement, to conventional antibiotics but new, safe and efficient means are needed for formulation and administration of the peptides. In this study we have investigated the utility of a novel type of lipid particles, the polyethylene glycol-stabilized lipid disks, as carriers for the model peptide melittin. The structural integrity of the carrier particle when loaded with the peptide was investigated using cryo-transmission electron microscopy. Liposome leakage upon addition of the peptide-lipid disks was monitored as a means to verify the membrane lytic effect of the formulation. The susceptibility of melittin to tryptic digestion was studied and compared in the absence and presence of lipid disks. Finally, the antibacterial effect of the peptide-lipid disk formulation was compared to that of free melittin after both single and repeated exposure to Escherichia coli. The results show that melittin can redistribute from the disk into a new host membrane and that formulation in the disks does not compromise melittin's membrane permeabilizing ability. Further, the peptide was found to be fully protected against degradation when bound to the disks. Time-kill experiments revealed that all the antibacterial effect of melittin administered in free form was gone after a single exposure to E. coli. In contrast, the disk formulation showed significant cell-killing effect also upon a second exposure to bacteria, indicating an extended release of peptide from the lipid disks. These results suggest that the lipid disks constitute a new class of promising carriers for peptide antibiotics.

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“…However, these benefits may be partially offset by a decrease in bioactivity because PEG-linked protein may interact less efficiently with its specific receptor molecule. A number of proteins have been pegylated, including asparaginase, tumor necrosis factor (TNF), human megacaryocyte growth and development factor (MGDF), interferon, brain-derived neurotrophic factor (BDNF), and interleukin-6 (IL-6) [11][12][13][14][15][16].…”

Section: Introductionmentioning

confidence: 99%

Pharmacodynamics and pharmacokinetics of single doses of subcutaneous pegylated human G‐CSF mutant (Ro 25‐8315) in healthy volunteers: Comparison with single and multiple daily doses of filgrastim

Auwera

Platzer

et al. 2001

American J Hematol

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Positional Isomers of Monopegylated Interferon α-2a: Isolation, Characterization, and Biological Activity

Cited by 109 publications

References 12 publications

Site of Pegylation and Polyethylene Glycol Molecule Size Attenuate Interferon-α Antiviral and Antiproliferative Activities through the JAK/STAT Signaling Pathway

Site of Pegylation and Polyethylene Glycol Molecule Size Attenuate Interferon-α Antiviral and Antiproliferative Activities through the JAK/STAT Signaling Pathway

PEG-stabilized lipid disks as carriers for amphiphilic antimicrobial peptides

Pharmacodynamics and pharmacokinetics of single doses of subcutaneous pegylated human G‐CSF mutant (Ro 25‐8315) in healthy volunteers: Comparison with single and multiple daily doses of filgrastim

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