Linear and Branched Bicin Linkers for Releasable PEGylation of Macromolecules: Controlled Release in Vivo and in Vitro from Mono- and Multi-PEGylated Proteins

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• 作者：Hong Zhao ; Karen Yang ; Anthony Martinez ; Amartya Basu ; Ramesh Chintala ; Hsien-Ching Liu ; Ahsen Janjua ; Maoliang Wang ; and David Filpula
• 刊名：Bioconjugate Chemistry
• 出版年：2006
• 出版时间：March 2006
• 年：2006
• 卷：17
• 期：2
• 页码：341 - 351
• 全文大小：179K
• 年卷期：v.17,no.2(March 2006)
• ISSN：1520-4812

文摘

The utility of PEGylation for improving therapeutic protein pharmacology would be substantially expanded if theauthentic protein drugs could be regenerated in vivo. Diminution of kinetic constants of both enzymes and proteinligands are commonly encountered following permanent bioconjugation with poly(ethylene glycol) polymers. Infurther development of releasable linker technology, we investigated an amino PEG anchimeric prodrug system,based on either the linear or branched bicin3 (BCN3) linkage, one promising representative of several aliphaticester structures synthesized from N-modifed bis-2-hydroxyethylglycinamide (bicin). Protein models included anenzyme, lysozyme, and a receptor ligand, interferon-beta-1b, for preparation of linear or branched mono- andmulti-PEGylated conjugates as inactive PEG-BCN3 prodrugs. The kinetics of protein release, both in plasma (invitro) and in mice (in vivo), correlated with the number of PEG attachments, and the plasma half-lives of PEGrelease spanned a duration of hours to days within the therapeutically relevant window. Capillary electrophoresis,SDS-PAGE, mass determination, and enzymatic and antiviral activity determinations demonstrated regenerationof equivalent native proteins from the inactive PEG-BCN3 conjugates. Pharmacokinetic analysis of the PEGylatedinterferon-beta-1b administered subcutaneously in mice demonstrated an over 20-fold expansion of the area underthe curve exposure of bioactive protein when compared to native protein.