摘要
一类具有潜在治疗不同社会相关人类疾病的药物以核酸为基础的药物(NABD)作为代表,包括小干扰RNA (siRNA)、Decoy寡核苷酸(decoy ODN) 、反义寡核苷酸 (ASOs)。虽然nABD可以设计专门针对几乎任何靶点,但他们对核酸酶降解的敏感性、传递到目标组织的困难性严重限制了它们的临床应用,这就需要建立一个适当的纳米结构传递系统。对于NABD的传递,水溶性和生物相容性多糖Inuli由精胺衍生而来,精胺是拥有四个胺基的柔性分子,pKa值的范围在8-11,pH值为7.4时主要质子化形式存在。相关共聚物(Inu-Spm))是通过使用一种称为增强微波合成(EMS) 的二步反应进行,与传统的微波反应比,EMS在反应容器内控制微波辐射的同时控能利用冷却气体控制温度,其有着更多的能量可以应用于反应系统。利用FT-IR光谱和1H-NMR对合成的菊粉衍生物进行鉴定。INU-Spm 衍生物的衍生度约为14% mol/mol。对这些结构进行了测试,评价其形成有着遗传物质(人工合成多聚物)的非共价复合物的能力。琼脂糖凝胶阻滞实验表明,合成的聚合物与DNA相互作用,并能够形成在不同C / P重量比的复合物。而且,用于分析聚合物形态大小和z电位的光散射研究表明,共聚物能与形成纳米体系的遗传物质相互作用。此外,16HBE细胞毒性试验证明了该复合物的生物相容性。利用siRNA使荧光素酶表达沉默的转染研究表明荧光素酶的表达减少约70%,这证明了共聚物转染同一细胞系的效率。这些研究结果鼓励我们利用这些共聚物来获得基于nABD传递系统的有效、可行的菊粉。
关键词: 菊粉,微波,核酸基础药物,聚合物,siRNA传递,精胺。
Current Drug Targets
Title:Inulin Derivatives Obtained Via Enhanced Microwave Synthesis for Nucleic Acid Based Drug Delivery
Volume: 16 Issue: 14
Author(s): Carla Sardo, Emanuela Fabiola Craparo, Calogero Fiorica, Gaetano Giammona and Gennara Cavallaro
Affiliation:
关键词: 菊粉,微波,核酸基础药物,聚合物,siRNA传递,精胺。
摘要: A new class of therapeutic agents with a high potential for the treatment of different socially relevant human diseases is represented by Nucleic Acid Based Drugs (NABD), including small interfering RNAs (siRNA), decoy oligodeoxynucleotides (decoy ODN) and antisense oligonucleotides (ASOs). Although NABD can be engineered to be specifically directed against virtually any target, their susceptibility to nuclease degradation and the difficulty of delivery into target tissues severely limit their use in clinical practice and require the development of an appropriate nanostructured delivery system. For delivery of NABD, Inulin (Inu), a natural, water soluble and biocompatible polysaccharide, was derivatized by Spermine (Spm), a flexible molecule with four amine groups that, having pKa values in the range between 8-11, is mainly in the protonated form at pH 7.4. The synthesis of related copolymers (Inu-Spm) was performed by a two step reaction, using a method termed Enhanced Microwave Synthesis (EMS) which has the advantage, compared to conventional microwave reaction, that high amount of energy can be applied to the reaction system, by administering microwave irradiation and simultaneously controlling the temperature in the reaction vessel with cooled air. The synthesized inulin derivatives were characterized by FT-IR spectra and 1H-NMR. INU-Spm derivatives with a degree of derivatization of about 14 % mol/mol were obtained. These polycations were tested to evaluate their ability to form non covalent complexes with genetic material (polyplexes). Agarose gel retardation assays showed that the obtained copolymers are able to electrostatically interact with DNA duplex to form polyplexes at different c/p weight ratios. Moreover, light scattering studies, performed to analyze size and z-potential of polyplexes, evidenced that copolymers are able to interact with genetic material leading to the formation of nanoscaled systems. In addition, biocompatibility of polyplexes was demonstrated by performing cytotoxicity assays on a 16HBE cell line. Transfection studies, performed by using siRNA able to silence luciferase expression, demonstrate the efficiency of polyplexes to transfect the same cell line, with a reduction of luciferase expression to about 70%. These results encourage us to work with these copolymers to obtain an efficient and feasible inulin based NABD delivery system.
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Carla Sardo, Emanuela Fabiola Craparo, Calogero Fiorica, Gaetano Giammona and Gennara Cavallaro , Inulin Derivatives Obtained Via Enhanced Microwave Synthesis for Nucleic Acid Based Drug Delivery, Current Drug Targets 2015; 16 (14) . https://dx.doi.org/10.2174/138945011614151119130426
DOI https://dx.doi.org/10.2174/138945011614151119130426 |
Print ISSN 1389-4501 |
Publisher Name Bentham Science Publisher |
Online ISSN 1873-5592 |
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