Abstract
Insulin mucoadhesive buccal films (MBF) are a noninvasive insulin delivery system that offers an advantageous alternative route of administration to subcutaneous injection. One major concern in the formulation of insulin MBF is the preservation of an insulin secondary structure in the presence of the other film components. Buccal films were formulated using chitosan, glycerin, and L-arginine. The MBF-forming solutions (MBF-FS) and the films (MBF) were examined for their chemical and structural stability and for their in vivo activity. Enzyme-Linked Immunosorbent Assay (ELISA) of the insulin-loaded MBF showed that each individualized unit dose was at least loaded with 80% of the insulin theoretical dose. Results of Synchrotron Radiation Circular Dichroism (SRCD) measurements revealed that MBF-FS retained the α-helices and β–sheets conformations of insulin. Fourier transform infrared (FTIR)-microspectroscopy (FTIR-MS) examination of insulin MBF revealed the protective action of L-arginine on insulin structure by interacting with chitosan and minimizing the formation of an unordered structure and β-strand. A blood glucose-lowering effect of insulin MBF was observed in comparison with subcutaneous (S.C) injection using a rat model. As a result; chitosan-based MBFs were formulated and characterized using SRCD and FTIR-MS techniques. Furthermore, the results of in vivo testing suggested the MBFs as a promising delivery system for insulin.
Highlights
Insulin is the main therapeutic agent for the treatment for type I diabetes and many advanced cases of type 2 patients [1]
The present study aims to examine the secondary structure of insulin loaded in MBFFS and mucoadhesive buccal films (MBF) using in vitro techniques; mainly Synchrotron radiation circular dichroism (SRCD) and Fourier transform infrared (FTIR)-MS in addition to the in vivo evaluation of insulin activity in the MBF using a rat model
The MBF-forming solution (MBF-FS) was prepared in two steps: the CH13,000 Da solution, which consists of 3% chitosan 13,000 Da, 30% glycerin, and 10% L-arginine was dissolved in 2% acetic acid using the homogenizer and mixed for 2 to 3 min followed by sonication for up to 5 min
Summary
Insulin is the main therapeutic agent for the treatment for type I diabetes and many advanced cases of type 2 patients [1]. Changes in insulin secondary structure can be examined using spectroscopy methods such as circular dichroism and infrared spectroscopy. The increased intensity of a synchrotron radiation light source makes it possible to obtain higher signal-to-noise ratios implying smaller sample requirements, in addition to the rapid measurements and the possibility of examining samples in the presence of high concentrations of buffers and other absorbing components [21]. Another biophysical technique; infrared spectroscopy, offers a valuable tool for studying the secondary structures of proteins, in the solid state. The present study aims to examine the secondary structure of insulin loaded in MBFFS and MBF using in vitro techniques; mainly SRCD and FTIR-MS in addition to the in vivo evaluation of insulin activity in the MBF using a rat model
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