Biochemical and biopharmaceutical properties of PEGylated uricase

Autor(es)

Freitas, Debora da Silva ^[1] ; Spencer, Patrick Jack ^[2] ; Vassao, Ruth Camargo ^[3] ; Abrahao-Neto, Jose ^[1]

Afiliação do(s) autor(es)

^[1] Univ Sao Paulo, Sch Pharmaceut Sci, Dept Biochem & Pharmaceut Technol, BR-05508000 Sao Paulo - Brazil

^[2] IPEN CNEN SP, Ctr Biotecnol, BR-05508000 Sao Paulo - Brazil

^[3] Inst Butantan, Biol Lab, BR-05503900 Sao Paulo - Brazil

• Tipo de documento: Artigo Científico
• Fonte: International Journal of Pharmaceutics; v. 387, n. 1-2, p. 215-222, MAR 15 2010.
• Citações Web of Science: 38

Resumo

PEGylation is a successful strategy for improving the biochemical and biopharmaceutical properties of proteins and peptides through the covalent attachment of polyethylene glycol chains. In this work, purified recombinant uricase from Candida sp. (UC-r) was modified by PEGylation with metoxypolyethilenoglycol-p-nitrophenyl-carbonate (mPEG-pNP) and metoxypolyethyleneglycol-4,6-dichloro-s-triazine (mPEG-CN). The UC-r-mPEG-pNP and UC-r-mPEG-CN conjugates retained 87% and 75% enzyme activity respectively. The K(M) values obtained 2.7 x 10(-5) M (mPEG-pNP) or 3.0 x 10(-5) M (mPEG-CN) lot the conjugates as compared to 5.4 x 10(-5) M for the native UC-r, suggesting enhancement in the substrate affinity of the enzyme attached. The effects of pH and temperature on PEGylated UC-r indicated that the conjugates were more active at close physiological pH and were stable up to 70 degrees C. Spectroscopic study performed by circular dichroism at 20 degrees C and 50 degrees C did not show any relevant difference in protein structure between native and PEGylated UC-r. In rabbit and Balb/c mice, the native UC-r elicited an intense immune response being highly immunogenic. On the other hand, the PEGylated UC-r when injected chronically in mice did not induce any detectable antibody response. This indicates sufficient reduction of the immunogenicity this enzyme by mPEG-pNP or mPEG-CN conjugation, making it suitable for a possible therapeutical use. (C) 2009 Elsevier B.V. All rights reserved. (AU)

• Processo FAPESP: 07/57457-1 - Funcionalidade e caracterização das propriedades físico-químicas, biológicas e estruturais da uricase modificada por peglação
• Beneficiário: Jose Abrahao Neto
• Modalidade de apoio: Auxílio à Pesquisa - Regular