Rehman Asad Ur, Busignies Virginie, Coelho Silva Ribeiro Marcela, Almeida Lage Nayara, Tchoreloff Pierre, Escriou Virginie, Charrueau Christine
Université de Paris, CNRS, INSERM, UTCBS, F-75006 Paris, France.
Univ. Bordeaux, CNRS, Arts et Metiers Institute of Technology, Bordeaux INP, INRAE, I2M Bordeaux, F-33400 Talence, France.
Pharmaceutics. 2021 Oct 28;13(11):1807. doi: 10.3390/pharmaceutics13111807.
The incorporation of siRNA into nanocarriers is mandatory to facilitate its intracellular delivery, as siRNA itself cannot enter cells. However, the incorporation of these nanocarriers into oral, solid dosage forms and their fate in the gastrointestinal environment is yet to be explored. In the present work, the fate of, (i) naked siRNA, (ii) freshly prepared siRNA lipoplexes, and (iii) tableted siRNA lipoplexes, in simulated gastric and intestinal fluids was studied. The siRNA, either released from or protected within the lipoplexes, was quantified by gel electrophoresis and siRNA efficacy was assessed in cell transfection. The freshly prepared lipoplexes kept their siRNA load and transfection efficiency totally preserved during 1 h of incubation in simulated gastric fluid at 37 °C. However, in simulated intestinal fluid, despite no release of siRNA from lipoplexes after 6 h of incubation, gene silencing efficacy was dramatically decreased even after 1 h of exposure. The lipoplexes obtained from tablets efficiently protected siRNA in simulated gastric fluid, thus preserving the gene silencing efficacy, whereas their incubation in simulated intestinal fluid resulted in a marked siRNA release and decreased gene silencing efficacy. These results provided a detailed explanation for understanding the fate of siRNA in gastrointestinal conditions, when simply loaded in lipoplexes or formulated in the form of tablets.
由于小干扰RNA(siRNA)自身无法进入细胞,因此将其包裹于纳米载体中以促进其细胞内递送是必不可少的。然而,将这些纳米载体引入口服固体剂型及其在胃肠道环境中的命运仍有待探索。在本研究中,考察了(i)游离siRNA、(ii)新制备的siRNA脂质体复合物以及(iii)制成片剂的siRNA脂质体复合物在模拟胃液和肠液中的命运。通过凝胶电泳对从脂质体复合物中释放或受其保护的siRNA进行定量,并在细胞转染实验中评估siRNA的有效性。新制备的脂质体复合物在37℃的模拟胃液中孵育1小时期间,其siRNA负载量和转染效率完全保持。然而,在模拟肠液中,尽管孵育6小时后脂质体复合物中未释放出siRNA,但即使仅暴露1小时,基因沉默效果也显著降低。片剂中的脂质体复合物在模拟胃液中能有效保护siRNA,从而保持基因沉默效果,而在模拟肠液中孵育则导致显著的siRNA释放和基因沉默效果降低。这些结果为理解siRNA在胃肠道条件下(简单负载于脂质体复合物中或制成片剂形式)的命运提供了详细解释。