Improving the therapeutic efficacy of peptides and proteins: A role for polysialic acids

Gregoriadis, Gregory; Jain, Sanjay; Papaioannou, Ioannis; Laing, Peter

doi:10.1016/j.ijpharm.2005.06.007

Cited by 177 publications

(118 citation statements)

References 12 publications

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“…The sialylation of proteins is known to prolong their half-life in vivo [21,22]. To examine whether this was true for C2del, the wild-type MFG-E8 and C2del proteins were injected into C57BL/6 mice, and their levels in serum were monitored by ELISA.…”

Section: C2del Binds To Phosphatidylserine and Enhances The Phagocytomentioning

confidence: 99%

Aberrant splicing of the milk fat globule‐EGF factor 8 (MFG‐E8) gene in human systemic lupus erythematosus

et al. 2010

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Milk fat globule-EGF factor 8 (MFG-E8) promotes the phagocytosis of apoptotic cells by serving as a bridging molecule between apoptotic cells and phagocytes. Many apoptotic cells are left unengulfed in the germinal centers of the spleen of MFG-E8 À/À mice, which develop a human systemic lupus erythematosus (SLE)-like autoimmune disease. Here, we analyzed the MFG-E8 gene in human SLE patients, and found in two out of 322 female patients a heterozygous intronic mutation, which caused a cryptic exon from intron 6 to be included in the transcript. The cryptic exon contained a premature termination codon, generating a C-terminally truncated MFG-E8 protein. The mutant MFG-E8 was aberrantly glycosylated and sialylated, but bound to phosphatidylserine and enhanced the phagocytosis of apoptotic cells. When intravenously injected into mice, the mutant MFG-E8 was sustained longer in the blood circulation than wild-type MFG-E8. Repeated administrations of the mutant MFG-E8 protein induced the production of autoantibodies, such as anti-cardiolipin and anti-nuclear antibodies, at a lower dose than that required for the wild-type protein. These results suggested that the intronic mutation in the human MFG-E8 gene can lead to the development of SLE.Key words: Aberrant splicing . Apoptosis . Autoimmiune disease . Protein stability IntroductionUnnecessary or harmful cells are eliminated by apoptosis, a form of programmed cell death in which cysteine proteases of the caspase family are activated to cleave cellular substrates [1,2]. The apoptotic cells are rapidly engulfed and digested by phagocytes such as macrophages and immature dendritic cells.The swift engulfment of cell corpses by phagocytes prevents the release of noxious or immunogenic debris from dying cells into the circulation. In the process of apoptosis, the dying cells expose phosphatidylserine on their external membrane in a caspasedependent manner. This externalization of phosphatidylserine is 1778one of the hallmarks of apoptosis and acts as an ''eat me'' signal for phagocytes [3]. Recently, several molecules that recognize phosphatidylserine have been identified [4][5][6][7].Systemic lupus erythematosus (SLE) is a chronic autoimmune disease caused by multiple genetic and environmental factors [8].Patients with SLE develop a broad spectrum of clinical manifestations affecting the skin, kidney, lungs, blood vessels, and/or nervous system. SLE is also characterized by the presence in sera of autoantibodies against nuclear components (anti-RNP and anti-DNA antibodies). Unengulfed apoptotic cells can be found in the germinal centers of the lymph nodes of some SLE patients, and macrophages from these patients show a reduced ability to engulf apoptotic cells [9]. Furthermore, circulating DNA or nucleosomes can also be found in the sera of SLE patients [10,11]. These results suggest that a deficiency in the clearance of apoptotic cells is one of the causes of SLE.Milk fat globule-EGF factor 8 (MFG-E8) is a glycoprotein. At the N-terminus, it has a EGF-like repeat(s), and...

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Section: C2del Binds To Phosphatidylserine and Enhances The Phagocytomentioning

confidence: 99%

Aberrant splicing of the milk fat globule‐EGF factor 8 (MFG‐E8) gene in human systemic lupus erythematosus

et al. 2010

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“…The chemical addition of polysialic acid (PSA) to proteins may represent another means to improve the circulating half-life of proteins, without causing adverse effects on the patient (6). In mammals, PSA is found mostly on the brain-specific neural cell adhesion molecule (NCAM) protein (7), on CD36, and on the recently described synaptic cell adhesion molecule (SynCAM) (8,9).…”

mentioning

confidence: 99%

Site-specific enzymatic polysialylation of therapeutic proteins using bacterial enzymes

Lindhout

Iqbal

Willis

et al. 2011

Proc. Natl. Acad. Sci. U.S.A.

108

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The posttranslational modification of therapeutic proteins with terminal sialic acids is one means of improving their circulating half-life, thereby improving their efficiency. We have developed a two-step in vitro enzymatic modification of glycoproteins, which has previously only been achieved by chemical means [Gregoriadis G, Jain S, Papaioannou I, Laing P (2005) Int J Pharm 300:125-130). This two-step procedure uses the Campylobacter jejuni Cst-II α2,8-sialyltransferase to provide a primer on N-linked glycans, followed by polysialylation using the Neisseria meningitidis α2,8-polysialyltransferase. Here, we have demonstrated the ability of this system to modify three glycoproteins with varying N-linked glycan compositions: the human therapeutic proteins alpha-1-antitrypsin (A1AT) and factor IX, as well as bovine fetuin. The chain length of the polysialic acid addition was optimized by controlling reaction conditions. After demonstrating the ability of this system to modify a variety of proteins, the effect of polysialylation on the activity and serum half-life of A1AT was examined. The polysialylation of A1AT did not adversely affect its in vitro inhibition activity against human neutrophil elastase. The polysialylation of A1AT resulted in a significantly improved pharmacokinetic profile when the modified proteins were injected into CD-1 mice. Together, these results suggest that polysialylated A1AT may be useful for improved augmentation therapy for patients with a deficiency in this protein and that this modification may be applied to other therapeutic proteins.glycosyltransferase | glycosylation

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“…for injection into humans in significant amounts for pretreatment against OPs with remarkable reduction of side effects, including immune response (34,38,39).…”

Section: Resultsmentioning

confidence: 99%

Chemical polysialylation of human recombinant butyrylcholinesterase delivers a long-acting bioscavenger for nerve agents in vivo

Ilyushin

Смирнов

Belogurov

et al. 2013

Proc. Natl. Acad. Sci. U.S.A.

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The creation of effective bioscavengers as a pretreatment for exposure to nerve agents is a challenging medical objective. We report a recombinant method using chemical polysialylation to generate bioscavengers stable in the bloodstream. Development of a CHO-based expression system using genes encoding human butyrylcholinesterase and a proline-rich peptide under elongation factor promoter control resulted in self-assembling, active enzyme multimers. Polysialylation gives bioscavengers with enhanced pharmacokinetics which protect mice against 4.2 LD 50 of S-(2-(diethylamino)ethyl) O-isobutyl methanephosphonothioate without perturbation of long-term behavior.organophosphate | N-acetylneuraminic acid | Russian VX | pesticide | mass spectrometry

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Improving the therapeutic efficacy of peptides and proteins: A role for polysialic acids

Cited by 177 publications

References 12 publications

Aberrant splicing of the milk fat globule‐EGF factor 8 (MFG‐E8) gene in human systemic lupus erythematosus

Aberrant splicing of the milk fat globule‐EGF factor 8 (MFG‐E8) gene in human systemic lupus erythematosus

Site-specific enzymatic polysialylation of therapeutic proteins using bacterial enzymes

Chemical polysialylation of human recombinant butyrylcholinesterase delivers a long-acting bioscavenger for nerve agents in vivo

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