Synthesis and Characterization of Peptide-Chitosan Conjugates (PepChis) with Lipid Bilayer Affinity and Antibacterial Activity

Autor(es)

Costa Petrin, Thais H. ^[1] ; Fadel, Valmir ^[2] ; Martins, Danubia B. ^[2] ; Dias, Susana A. ^[3] ; Cruz, Ana ^[3] ; Sergio, Luciana Marciano ^[4] ; Arcisio-Miranda, Manoel ^[4] ; Castanho, Miguel A. R. B. ^[3] ; dos Santos Cabrera, Marcia P. ^{[2, 1]}

Afiliação do(s) autor(es)

^[1] Univ Estadual Paulista Unesp, Inst Biociencias Letras & Ciencias Exatas Ibilce, Dept Quim & Ciencias Ambientais, Campus Sao Jose Do Rio Preto, BR-15054000 Sao Jose Do Rio Preto, SP - Brazil

^[2] Univ Estadual Paulista Unesp, Inst Biociencias Letras & Ciencias Exatas Ibilce, Dept Fis, Campus Sao Jose Do Rio Preto, BR-15054000 Sao Jose Do Rio Preto, SP - Brazil

^[3] Univ Lisbon, Fac Med, Inst Med Mol, P-1649028 Lisbon - Portugal

^[4] Univ Fed Sao Paulo, Dept Biofis, Lab Neurobiol Estrutural & Func LaNEF, BR-04023062 Sao Paulo - Brazil

• Tipo de documento: Artigo Científico
• Fonte: Biomacromolecules; v. 20, n. 7, p. 2743-2753, JUL 2019.
• Citações Web of Science: 0

Resumo

Antimicrobial peptides appear among innovative biopolymers with potential therapeutic interest. Nevertheless, issues concerning efficiency, production costs, and toxicity persist. Herein, we show that conjugation of peptides with chitosans can represent an alternative in the search for these needs. To increase solubility, deacetylated and degraded chitosans were prepared. Then, they were functionalized via N-succinimidyl-S-acetylthiopropionate or via glutathione (GSH), an endogenous peptide linker. To the best of our knowledge, it is the first time that GSH is used as a thiolating agent for the conjugation of peptides. Next, thiolated chitosans were conjugated through a disulfide bond with designed short chain peptides, one of them derived from the antimicrobial peptide Jelleine-I. Conjugates and respective reaction intermediates were characterized by absorciometry, attenuated total reflectance-Fourier transform infrared, and H-1 NMR Zeta potential measurements showed the cationic nature of these biomacromolecules and their preferential partitioning to Gram-positive bacterial-like model membranes. In vitro investigation using representative Gram-positive and-negative bacteria (Staphylococcus aureus and Escherichia coli, respectively) showed that the conjugation strategies lead to enhanced activity in relation to the unconjugated peptide and to the unconjugated chitosan. The obtained products showed selectivity toward S. aureus at low cytotoxicity as determined in NIH/3T3 cells. Overall, our study demonstrates that an appropriate choice of antimicrobial peptide and chitosan characteristics leads to increased antimicrobial activity of the conjugated product and represents a strategy to modulate the activity and selectivity of antimicrobials resorting to low-cost chemicals. The present proposal starts from less expensive raw materials (chitosan and short-chain peptide), is based on aqueous solvents, and minimizes the use of reactants with a higher environmental impact. The final biopolymer contains the backbone of chitosan, just 3-6% peptide derived from royal jelly and GSH, all of them considered safe for human use or as a physiological molecule. (AU)

• Processo FAPESP: 14/08372-7 - Moléculas conjugadas peptídeo/quitosana com potencial farmacologico: síntese, prospecção de atividade em sistemas mimeticos de membrana e avaliações em células
• Beneficiário: Marcia Perez dos Santos Cabrera
• Modalidade de apoio: Bolsas no Brasil - Jovens Pesquisadores
• Processo FAPESP: 16/13368-4 - Sistemas nanoestruturados: de modelos biomiméticos de membranas a carreadores de bioativos
• Beneficiário: Karin Do Amaral Riske
• Modalidade de apoio: Auxílio à Pesquisa - Temático
• Processo FAPESP: 15/07548-7 - Nova rota para síntese de conjugados peptideo-quitosana: efeitos da hidrofobicidade do peptídeo avaliados na interação com membranas modelo via potencial zeta
• Beneficiário: Thais Helena Costa Petrin
• Modalidade de apoio: Bolsas no Brasil - Iniciação Científica
• Processo FAPESP: 12/02065-0 - Eletrofisiologia de peptídeos antimicrobianos em bicamadas lipídicas planas
• Beneficiário: Manoel de Arcisio Miranda Filho
• Modalidade de apoio: Auxílio à Pesquisa - Regular
• Processo FAPESP: 12/24259-0 - Moléculas conjugadas peptídeo/quitosana com potencial farmacológico: síntese, prospecção de atividade em sistemas miméticos de membrana e avaliações em células
• Beneficiário: Marcia Perez dos Santos Cabrera
• Modalidade de apoio: Auxílio à Pesquisa - Jovens Pesquisadores