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 woundhealing properties.
Methods
In this study, we analyzed the association between β-cyclodextrin (βCD) 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 βCD. Higher secondary interactions, namely hydrophobic interactions, hydrogen bonding and van der Waals forces were observed at higher P1G10: βCD mass ratios. These results provide evidence of the occurrence of strong solid-state supramolecular non-covalent interactions between P1G10 and βCD. Microcalorimetric analysis demonstrates that complexation results in a favorable enthalpic contribution, as has already been described during formation of similar βCD 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 βCDs as candidates for complexing cysteine proteinases, resulting in supramolecular arrays with sustained proteolytic activity.
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Acknowledgments
The authors acknowledge the financial support of the Brazilian agencies CAPES, CNPq, PRONEX-FAPEMIG-CNPq, and Rede Nanobiotec-CNPq and Dr Abraham Schnaiderman. We thank Dr Adelaide Faljoni Alario of IQ-USP for allowing us access to the CD equipment.
The authors have no conflicts of interest that are directly relevant to the content of this study.
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Denadai, Â.M.L., Santoro, M.M., Lopes, M.T.P. et al. A Supramolecular Complex between Proteinases and β-Cyclodextrin that Preserves Enzymatic Activity. BioDrugs 20, 283–291 (2006). https://doi.org/10.2165/00063030-200620050-00004
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DOI: https://doi.org/10.2165/00063030-200620050-00004