Monoconjugation of Human Amylin with Methylpolyethyleneglycol

Tháyna Sisnande; Luiz Henrique Guerreiro; Raquel R Braga; Luana Jotha-Mattos; Luiza C S Erthal; Priscilla Tinoco; Bruno M Ferreira; Luís Maurício T R Lima

doi:10.1371/journal.pone.0138803

Monoconjugation of Human Amylin with Methylpolyethyleneglycol

PLoS One. 2015 Oct 8;10(10):e0138803. doi: 10.1371/journal.pone.0138803. eCollection 2015.

Authors

Tháyna Sisnande¹, Luiz Henrique Guerreiro², Raquel R Braga¹, Luana Jotha-Mattos¹, Luiza C S Erthal¹, Priscilla Tinoco², Bruno M Ferreira¹, Luís Maurício T R Lima³

Affiliations

¹ Federal University of Rio de Janeiro-UFRJ, CCS, Bss24, Ilha do Fundão, 21941-590, Rio de Janeiro, RJ, Brazil.
² Department of Chemistry, Institute of Exact Sciences, Rural Federal University of Rio de Janeiro-Universidade Federal Rural do Rio de Janeiro, Rodovia BR 465, km 7, CEP: 23890-000, Seropédica, RJ, Brazil.
³ Federal University of Rio de Janeiro-UFRJ, CCS, Bss24, Ilha do Fundão, 21941-590, Rio de Janeiro, RJ, Brazil; Laboratory for Macromolecules (LAMAC-DIMAV), Brazilian National Institute of Metrology, Quality and Technology-INMETRO, Av. N. Sa. das Graças, 50-Xerém, Duque de Caxias-RJ, 25250-020, Rio de Janeiro, Brazil; National Institute of Science and Technology for Structural Biology and Bioimaging (INBEB-INCT), Federal University of Rio de Janeiro, Rio de Janeiro 21941-590, Brazil.

Abstract

Amylin is a pancreatic hormone cosecreted with insulin that exerts unique roles in metabolism and glucose homeostasis. The therapeutic restoration of postprandial and basal amylin levels is highly desirable in diabetes mellitus. Protein conjugation with the biocompatible polymer polyethylene glycol (PEG) has been shown to extend the biological effects of biopharmaceuticals. We have designed a PEGylated human amylin by using the aminoreactive compound methoxylpolyethylene glycol succinimidyl carbonate (mPEGsc). The synthesis in organic solvent resulted in high yields of monoPEGylated human amylin, which showed large stability against aggregation, an 8 times increase in half-life in vivo compared to the non-conjugated amylin, and pharmacological activity as shown by modulation of cAMP production in MCF-7 cell line, decrease in glucagon and modulation of glycemia following subcutaneous administration in mice. Altogether these data reveal the potential use of PEGylated human amylin for the restoration of fasting amylin levels.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Blood Glucose / drug effects
Cyclic AMP / metabolism
Glucagon / metabolism
Homeostasis / drug effects
Humans
Hypoglycemic Agents / chemistry
Hypoglycemic Agents / pharmacology
Islet Amyloid Polypeptide / chemistry*
Islet Amyloid Polypeptide / metabolism
Islet Amyloid Polypeptide / pharmacology
Kinetics
MCF-7 Cells
Male
Mice
Polyethylene Glycols / chemistry*
Polyethylene Glycols / pharmacology
Solvents / chemistry

Substances

Blood Glucose
Hypoglycemic Agents
Islet Amyloid Polypeptide
Solvents
Polyethylene Glycols
Glucagon
Cyclic AMP

Grants and funding

This research was supported by the Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES), Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq; www.cnpq.br; 403525/2012-8; 472779/2012-5), Fundação de Amparo à Pesquisa do Estado do Rio de Janeiro Carlos Chagas Filho (FAPERJ; www.faperj.br; E-26/103.260/2011; E-26-111.383/2010) and the BioZeus Desenvolvimento de Produtos Biofarmacêuticos S.A. (for financial support for the cAMP assay). The funding agencies had no role in the study design, data collection and analysis, or decision to publish, or prepare of the manuscript.