A supramolecular complex between proteinases and beta-cyclodextrin that preserves enzymatic activity: physicochemical characterization

Angelo M L Denadai, Marcelo M Santoro, Miriam T P Lopes, Angélica Chenna, Frederico B de Sousa, Gabriela Mol Avelar, Marco R Túlio Gomes, Fanny Guzman, Carlos E Salas, Rubén D Sinisterra

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Abstract

BACKGROUND: Cyclodextrins are suitable drug delivery systems because of their ability to subtly modify the physical, chemical, and biological properties of guest molecules through labile interactions by formation of inclusion and/or association complexes. Plant cysteine proteinases from Caricaceae and Bromeliaceae are the subject of therapeutic interest, because of their anti-inflammatory, antitumoral, immunogenic, and wound-healing properties.

METHODS: In this study, we analyzed the association between beta-cyclodextrin (betaCD) and fraction P1G10 containing the bioactive proteinases from Carica candamarcensis, and described the physicochemical nature of the solid-state self-assembled complexes by Fourier transform infrared (FTIR) spectroscopy, thermogravimetry (TG), differential scanning calorimetry (DSC), X-ray powder diffraction (XRD), and nuclear magnetic resonance (NMR), as well as in solution by circular dichroism (CD), isothermal titration calorimetry (ITC), and amidase activity.

RESULTS AND DISCUSSION: The physicochemical analyses suggest the formation of a complex between P1G10 and betaCD. Higher secondary interactions, namely hydrophobic interactions, hydrogen bonding and van der Waals forces were observed at higher P1G10 : betaCD mass ratios. These results provide evidence of the occurrence of strong solid-state supramolecular non-covalent interactions between P1G10 and betaCD. Microcalorimetric analysis demonstrates that complexation results in a favorable enthalpic contribution, as has already been described during formation of similar betaCD inclusion compounds. The amidase activity of the complex shows that the enzyme activity is not readily available at 24 hours after dissolution of the complex in aqueous buffer; the proteinase becomes biologically active by the second day and remains stable until day 16, when a gradual decrease occurs, with basal activity attained by day 29.

CONCLUSION: The reported results underscore the potential for betaCDs as candidates for complexing cysteine proteinases, resulting in supramolecular arrays with sustained proteolytic activity.

Original languageEnglish
Pages (from-to)283-91
Number of pages9
JournalBioDrugs
Volume20
Issue number5
Publication statusPublished - 2006

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Keywords

  • Amidohydrolases
  • Calorimetry
  • Calorimetry, Differential Scanning
  • Carica
  • Circular Dichroism
  • Latex
  • Magnetic Resonance Spectroscopy
  • Peptide Hydrolases
  • Spectroscopy, Fourier Transform Infrared
  • Thermogravimetry
  • X-Ray Diffraction
  • beta-Cyclodextrins

Cite this

Denadai, A. M. L., Santoro, M. M., Lopes, M. T. P., Chenna, A., de Sousa, F. B., Mol Avelar, G., ... Sinisterra, R. D. (2006). A supramolecular complex between proteinases and beta-cyclodextrin that preserves enzymatic activity: physicochemical characterization. BioDrugs, 20(5), 283-91.