Nanofibrous Formulation of Cyclodextrin Stabilized Lipases for Efficient Pancreatin Replacement Therapies

The purpose of enzyme therapies (ERT) is simple: patients who suffer from enzyme deficiency should be treated by administering enzymes from external sources. Since the introduction of these therapies, one of the main objectives was, and still is, to provide patients, who suffer from the lack of digestive enzymes with a per os formula, that contains proteins, which are capable of overtaking the role of the absent main digestive enzymes (lipase, protease and amylase) in the breakdown of nutrients. Naturally, the production of such preparations puts the pharmacists in front of unique challenges. Cyclodextrins (CD) are known to be excellent for the complexation of small molecules and consequently influencing their physical and chemical properties. Thanks to this and their atoxic nature, CDs are widely used in the pharmaceutical industry as additives. Research has shown that CDs are capable of developing specific interactions not only with small molecules, but also with macromolecules such as proteins. Thanks to these diverse interactions, CDs are able to enhance the activity and stability enzymes. As a result, CDs are considered as promising additives for ERT formulations. In this research, the authors aimed to develop a drug formulation containing cyclodextrin-stabilized lipases for the treatment of exocrine pancreatic insufficiency, a complication of chronic pancreatitis. The chosen formulation method was electrostatic fiber formation, which was used to immobilize the lipase-CD pairs in polyvinyl alcohol (PVA) nanofibers. During the research, the hydrolytic activity of 2 lipases (lipase form Aspergillus oryzae and lipase from Burkholderia cepacia) was investigated in the presence of 5 different CDs (β-Cyclodextrin, 2-hydroxypropyl-β-cyclodextrin, 2-hydroxypropyl-γ-cyclodextrin, randomly methylated β-cyclodextrin, and sulfobutylated-β-Cyclodextrin), using p-nitrophenyl palmitate as the model substrate. Results showed that the quality and weight ratio of lipases and cyclodextrins significantly affected the enzyme activity. Using lipase-CD pairs with optimal weight ratios, the conversion achieved in the hydrolytic breakdown was multiplied. It was found, that immobilizing the lipase-CD pairs in PVA nanofibers caused a loss in the activity-enhancing effect of the CDs due to interactions between the CDs and the PVA carrier matrix. It was also discovered, that the presence of CDs helped in achieving homogeneous distribution of lipases in the PVA nanofibers.

Tóth Gergő Dániel; Kazsoki Adrienn;    Gyarmati Benjámin;    Szilágyi András;    Vasvári Gábor; Katona Gábor; Szente Lajos; Zelkó Romána;    Poppe László;    Balogh-Weiser Diána^*;    Balogh György T.^*: Nanofibrous Formulation of Cyclodextrin Stabilized Lipases for Efficient Pancreatin Replacement Therapies. Pharmaceutics13(7):972. doi: 10.3390/pharmaceutics13070972.