Effects of cryoprotectants on the viability and activity of freeze dried recombinant yeasts as novel oral drug delivery systems assessed by an artificial digestive system

被引:43
作者
Blanquet, S [1 ]
Garrait, G [1 ]
Beyssac, E [1 ]
Perrier, C [1 ]
Denis, S [1 ]
Hébrard, G [1 ]
Alric, M [1 ]
机构
[1] Univ Auvergne, Fac Pharm, ERT, CIDAM,CRNH, F-63001 Clermont Ferrand, France
关键词
oral formulations; pre-screening; freeze-drying; cryoprotectant; recombinant Saccharomyces cerevisiae; artificial digestive system;
D O I
10.1016/j.ejpb.2005.03.009
中图分类号
R9 [药学];
学科分类号
1007 ;
摘要
The aim of this study was to investigate, in a gastric-small intestinal system TIM-1, the effect of cryoprotectants on the survival of freeze-dried Saccharomyces cerevisiae expressing the heterologous P450 73A1 and their ability to convert trans-cinnamic acid into p-coumaric acid. Yeasts were lyophilized in suspensions of trehalose, maltose, lactose, or a milk proteins/trehalose mix. Freeze-dried or native yeasts and trans-cinnamic acid were introduced simultaneously into TIM-1 at the beginning of digestion. Yeast survival rate was evaluated by cell counting in the ileal effluents. P450 73A1 activity was followed by HPLC assay of p-coumaric acid. Freeze-dried yeasts showed high tolerance to digestive conditions. Nevertheless, their survival rate was lower than that of non-dried cells (around 80% whatever the protective agent vs. 96%). The ability of recombinant freeze-dried S. cerevisiae to perform a bioconversion reaction in the digestive tract was shown with all the protectants. The highest trans-cinnamic acid conversion rate (24 vs. 41% for native yeasts) was obtained with the milk proteins/trehalose mix. These results show that freeze-drying might be considered for the pharmaceutical formulation of new drug delivery systems based on orally administered recombinant yeasts and that TIM-1 could be a helpful tool for the pre-screening of oral dosage forms. (c) 2005 Elsevier B.V. All rights reserved.
引用
收藏
页码:32 / 39
页数:8
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