《药用高分子材料sirna药物ppt课件》由会员分享，可在线阅读，更多相关《药用高分子材料sirna药物ppt课件（42页珍藏版）》请在金锄头文库上搜索。
1、姚老师作业：聚乳酸的本体聚合工艺，相应机理和反应方程,第六章 高分子纳米药物-siRNA药物,病变组织的基因活性,正常表达,过度表达,低表达,Emulate Drug?,抗体: Bind protein, but only extracellular epitopes Low throughput, high cost反义RNA/核酶: Stop gene expression (production), most proteins Chemical stabilization essential Barriers: sequence selection, potency, toxicitie
2、s, RNA干扰: Stop gene expression (production), most proteins Good activity without chemical stabilization Rapidly replacing antisense: really better?,特异性基因下调手段,RNA干扰（RNA interference, RNAi）技术,功能：特异性地沉默内源或外源性靶基因影响：2001年Science杂志十大科学成就之二 2002年Science杂志十大科学成就之首医学领域的应用：基因功能研究；基因敲除；基因治 疗；基因表达调控等;抗病毒以及糖尿病、肥
3、胖症和癌症、 黄斑变性的眼科疾病治疗等。,RNAi 作用机制,Nature，2004，430：161,From H. Winkler, IBC Conference “RNA Interference”, San Diego, CA, Feb 2003,siRNA,dsRNA,siRNA,RNAse,shRNA,In Vitro,In Vivo,In Vivo,miRNA,Adapted From Dykxhoorn et al., Nature Reviews 4, 457, 2003,具有RNAi机制的药物,Target ID,siRNA Target Validation Growing
4、,Clinical,Lead Discovery & Optimization,Preclinical,siRNA药物: 革命性突破,Potential of siRNA-Based Therapy,Phase 1 Phase 2 Phase 3,RNA干扰疗法的潜力,siRNA药物迅速进入临床研究,难以被细胞摄取：大分子，阴离子,脱靶效应：非特异性沉默，引起免疫刺激等,难以富集在肿瘤部位：体内不稳定， 半衰期短,1,2,3,Potential of siRNA-Based Therapy,RNA干扰疗法的潜力,siRNA药物临床引用的关键,Nature 2010, 468, p487, No
5、v. 25, 2010,Potential of siRNA-Based Therapy,RNA干扰疗法的潜力,siRNA给药技术成为关键挑战,表达siRNA的质粒DNA的传递 病毒（Nature Biotechnology, 2002, 20: 1006-10） 非病毒类载体 化学合成的siRNA的传递非病毒类载体：脂质体和高分子物理手段: electroporation, high pressure/high volume injection,Potential of siRNA-Based Therapy,RNA干扰疗法的潜力,siRNA的传递：体内RNAi技术应用的关键,脂质类,高分子
6、纳米载体,抗体类,化学修饰siRNA,siRNA缀合物,Potential of siRNA-Based Therapy,RNA干扰疗法的潜力,siRNA的非病毒载体种类,DAN和siRNA传递的比较,化学修饰的siRNA,Nature，2004，432：173-178,化学修饰的siRNA,Challenge 1: 稳定 Challenge 2: 长循环 Challenge 3: 靶向 Challenge 4: 渗透 Challenge 5: 细胞摄取 Challenge 6: 制剂化,基于纳米载体的 siRNA给药系统,Engineering Nanoparticles for siRNA
7、 Delivery,Extended Circulation Tissue Targeting,Penetration,Cell internalization,脂质体类siRNA纳米载体,脂质体类siRNA纳米载体,Stable Nucleic Acid Lipid Particle (SNALP),Stable Nucleic Acid Lipid Particle (SNALP),Mean particle size: 14012 nm Mean encapsulation efficiency: 93%3% Nucleic acid/lipid ratio: 424g/mol,siRN
8、A-SNALP with a longer half-life in plasma and liver reduced serum HBV DNA titers in mouse model following intravenous injection,Nature Biotechnology 23, 1002 - 1007 (2005),ApoB-siRNA-SNALP resulted in silencing of ApoB mRNA expression in liver and significant reductions in ApoB protein, serum choles
9、terol and low-density lipoprotein levels,Nature 441, 111-114 (4 May 2006),高分子类 siRNA给药系统,Steric Coat (Inert),Active Core,Exposed Targeting Ligands,Cleavable Link (Shedding),Ligands,Steric polymer,+,siRNA Payload,水溶性阳离子高分子纳米载体:,NUCLEIC ACIDS RESEARCH 32 (19): Art. No. e149 2004,PNAS 2007, 104, 5715-5
10、721,靶向修饰的水溶性阳离子高分子纳米载体:,Cancer Research 2005, 65, 8984-8992,PEG,JACS, 127, 1624-1625,靶向修饰的水溶性高分子-siRNA键合物:,JACS, 126 (42), 13612 -13613,Loading siRNA post nanoparticle formation PEG: prolonged blood circulation Biocompatible Degradable, potentially intracellular siRNA release Co-delivery of chemothe
11、rapeutic drug for synergistic effect,亲水嵌段,疏水嵌段,带正电荷嵌段,阳离子胶束/siRNA复合纳米颗粒:,Sun TM, et al., ACS Nano, 2011, 5, 1483-1494,Days post-xenografting,12,16,20,24,28,32,36,40,44,Sun TM, et al., ACS Nano, 2011, 5, 1483-1494,-actin,Kim A.Woodrow, W. Mark Saltzman et al., Nature Materials, 2009, 8, 526 533 Chris
12、topher J. Cheng, W. Mark Saltzman, Biomaterials, 2011, 32, 61946203,siRNA包载效率低下，仅为21%！ 1 g siRNA目前的价格为200万！,基于聚乙二醇-聚乳酸的siRNA纳米给药体系,Organic phase (polymer/lipid),aqueous phase (siRNA),Such nanoparticles, which is based FDA approved PEG-PLA, exhibit high encapsulation efficiency of siRNA (about 96%).,
13、Removing the organic solvent,Second emulsion,More water,PEG-PLA/Cationic lipid/siRNA nanoparticles,First emulsion,Cationic Lipid Assisted Nanoparticles: CLAN,基于CLAN技术设计的siRNA给药系统,基于CLAN技术设计的靶向siRNA给药系统,Nature Biotechnol. 23, 709717 (2005),抗体类 siRNA给药系统,M Sioud1, Gene Therapy, 2006, 13, 194195,思考题,RNAi原理和siRNA siRNA给药载体有几种，分别是什么，并简单描述。 siRNA和质粒DNA的差别，它们的给药方式有何区别以及一致的地方。 siRNA和质粒DNA针对疾病治疗的原理有什么差异。,
