1、Chapter 21 Targeted Drug Delivery System(TDDS)掌握靶向给药系统的概念、分类、特点。熟悉靶向制剂的体内外质量评价。了解分子药剂学和细胞生物学技术，了解靶向制剂的药动学基础。熟悉被动靶向制剂中脂质体、纳米粒、微乳等的特点、组成、靶向原理及制备方法、质量评价。了解主动靶向制剂和物理化学靶向制剂的类型、特点.Drug delivery system(DDS)Drug delivery system(DDS)给药系统给药系统A drug delivery system(DDS)is defined as a formulation or a device t
2、hat enables the introduction of a therapeutic substance 治疗物治疗物质质in the body and improves its efficacy and safety by controlling the rate,time,and place of release of drugs in the body.This process includes the administration of the therapeutic product,the release of the active ingredients by the pro
3、duct,and the subsequent transport of the active ingredients across the biological membranes to the site of action.Definition Drug delivery system is an interface between the patient and the drug.It may be a formulation of the drug to administer it for a therapeutic purpose or a device used to delive
4、r the drug.Targeted drug delivery system(TDDS)Definition The selective delivery of a drug to a specific region(tissue,cell type)of the body经某种途径给药后，药物通过特殊载体的作经某种途径给药后，药物通过特殊载体的作用，用，特异性地浓集特异性地浓集在靶部位的给药系统。在靶部位的给药系统。The aim of targeted drug delivery is to improve the therapeutic effectiveness of a drug
5、(or treatment)while at the same time improving its safety.Therefore,the aim is to increase the therapeutic index of a drug.6Superiorities of TDDSTraditional drug delivery systems:Medication is distributed throughout the body through the systemic blood circulation;A small portion of the medication re
6、aches the organ to be affected.Superiorities of TDDSTargeted drug delivery:Concentrate the medication in the tissues of interest;Reducing the relative concentration of the medication in the remaining tissues SIDE EFFECTS EFFICACY Target position:an organ or tissuea particular cell typea specific int
7、racellular compartment.特定细胞器内特定细胞器内Classification of TDDS 分类分类Passive targeted drug delivery 被动靶向被动靶向Active targeted drug delivery 主动靶向主动靶向Physicochemical targeted drug delivery 物理化学靶向物理化学靶向Classification of TDDSClassification of TDDS体内靶向性评价体内靶向性评价(1)(1)Relative uptake rate 相对摄取率相对摄取率 R Re e=(AUC=(A
8、UCi i)p p/(AUC/(AUCi i)s s (2)(2)Peak concentration ratio 峰浓度比峰浓度比 C Ce e=(Cmax)p/(Cmax)s=(Cmax)p/(Cmax)s(3)(3)Targeting efficiency 靶向效率靶向效率 T Te e=(AUC)=(AUC)targettarget/(AUC)/(AUC)non-targetnon-target 传统的隔室药动学模型不适于评价靶向制剂传统的隔室药动学模型不适于评价靶向制剂Molecular PharmaceuticsMolecular Pharmaceutics从分子水平和细胞水平研究
9、剂型因素对药物疗效的从分子水平和细胞水平研究剂型因素对药物疗效的影响的科学。影响的科学。1)1)药物分子与药物分子与DDSDDS的体内外转运与代谢的体内外转运与代谢2)2)药物分子与载体的相互作用药物分子与载体的相互作用3)3)靶向给药的分子机制靶向给药的分子机制4)4)分子水平的药代动力学研究分子水平的药代动力学研究12Particulate drug carriers 微粒类载体微粒类载体YLiposome 脂质体脂质体YNanoparticles/macroparticles 纳米粒纳米粒/微球微球YMicelles 胶束胶束YEmulsion 乳剂乳剂YDendrimers 树突状物树
10、突状物LIPOSOMES 脂质体脂质体YWhat are LiposomesYAdvantages of LiposomesYClasses of LiposomesYPreparation of LiposomesLiposomes are phospholipid based vesicles comprising of aqueous and lipid compartments.脂质体是利用脂质体是利用磷脂磷脂双分子层膜所形成的囊泡。双分子层膜所形成的囊泡。Based on the solubility,a drug can be encapsulated either in the
11、aqueous compartment or in the lipid bilayer.药物分子可被包裹在水相或脂药物分子可被包裹在水相或脂质双层中。质双层中。HydrophilicHydrophobic Phospholipids are amphiphathic in nature,containing a hydro-phobic tail and hydrophilic head.磷脂为两性物质，胆固磷脂为两性物质，胆固醇亦属两亲物质，有疏水亲水两种基团醇亦属两亲物质，有疏水亲水两种基团.Types of the LiposomesImmunoCationicLong Circulat
12、ing Conventional TargetedDoxil-FDADoxil-FDA批准的第一个纳米药物批准的第一个纳米药物通用名：Doxorubicin Liposomal；阿霉素脂质体商品名：Doxil（多喜）,Caelyx(楷莱)公司：Sequus Pharmaceuticals 应用：最初用于治疗卡波氏肉瘤，后被批准用于卵巢癌治疗 Amphotericin B Liposome1 1）Targeting ability 靶向性和淋巴定向性靶向性和淋巴定向性 2 2）Extended circulation time 长效性长效性 3 3）Good bioacceptability a
13、nd biocompatibility 细胞亲和性与组织相容性细胞亲和性与组织相容性 4 4）Reduce the toxicity of crude drug 降低药物降低药物毒性毒性5 5）Increase the stability of drug 提高药物稳定性提高药物稳定性 物理和化学稳定性较差（磷脂分子的氧化物理和化学稳定性较差（磷脂分子的氧化)脂质体的剂型特点脂质体的剂型特点磷脂磷脂+胆固醇胆固醇a、磷脂类磷脂类 卵磷脂、脑磷脂、大豆磷脂和人工合成磷脂卵磷脂、脑磷脂、大豆磷脂和人工合成磷脂 中性磷脂、负电荷磷脂、正电荷磷脂、中性磷脂、负电荷磷脂、正电荷磷脂、长循环磷脂长循环磷脂b
14、 b、胆固醇、胆固醇 脂质体“流动性缓冲剂”(fluidity buffer)调节膜流动性调节膜流动性 制备脂质体的材料制备脂质体的材料Materials:Phospholipids+cholesterol Preparation of liposomePreparation of liposomeNeutral:PC,DPPC,DMPC,DOPENegatively-charged:PA,OG,DCPPositively-charged:SA,DDABCholesterol:fluidity buffer Phospholipids Common components of the lipo
15、somesCholesterol Cholesterol acts as a“fluidity buffer”.胆固醇擦入磷脂双分子层中，改变磷脂中疏水链的胆固醇擦入磷脂双分子层中，改变磷脂中疏水链的流动性。流动性。This adds rigidity刚性刚性 to the lipid bilayers.Preparation methodPreparation methodFilm dispersion method 薄膜法薄膜法High-pressure homogenizer method 高压乳匀法高压乳匀法Reverse phase evaporation vesicle metho
16、d 逆向蒸发法逆向蒸发法Injection method 注入法注入法Freeze-thawing method 冻融法冻融法Film dispersion methodFilm dispersion method1.Phospholipids and cholesterol were dissolved in organic solvents2.Remove organic solvents by rotary evaporation，to form uniform film3.Add water and keep stirringInjection methodInjection meth
17、od1.Dissolve phospholipids and cholesterol in organic solvents2.Inject the solvent into phosphate buffer（50-60）3.keep stirring until the organic solvent is removed1、Morphology,particle size and distribution 形态、粒径及其分布形态、粒径及其分布2、Entrapment efficiency and loading capacity 载药量和包封率载药量和包封率 包封率=脂质体中的药量/（介质
18、中的药量+脂质体中的药量）100%3、Leakage rate 渗漏率渗漏率 渗漏率=（储存一定时间后渗漏到介质中的药量/储存前包封的药量）100%4Distribution in vivo 药物体内分布药物体内分布 脂质体制剂的质量评价脂质体制剂的质量评价Nanoparticles Nanoparticles 纳米粒纳米粒Colloidal particles ranging in size between 1 and 1000nm are known as nanoparticles 由天然或合成的高分子载体材料制成的、粒度在纳由天然或合成的高分子载体材料制成的、粒度在纳米数量级（米数量级
19、（1-1000nm1-1000nm）的固态胶体微粒）的固态胶体微粒 Nanocapsule纳米囊属药纳米囊属药库膜壳型，由聚合材料外库膜壳型，由聚合材料外壳和液状核构成，药物主壳和液状核构成，药物主要溶解在液状核中，这种要溶解在液状核中，这种纳米囊主要适于包裹脂溶纳米囊主要适于包裹脂溶性药物。性药物。Nanosphere 纳米球属基质纳米球属基质骨架型，属于实心的球或微骨架型，属于实心的球或微粒，药物吸附在其表面或包粒，药物吸附在其表面或包裹、溶解在其内部。裹、溶解在其内部。Advantages of NP deliveryAdvantages of NP deliveryPhysical s
20、tability 物理稳定性好物理稳定性好Belongs to colloid system 属胶体系统，较混悬型微球制剂容属胶体系统，较混悬型微球制剂容易给药易给药Extended therapeutic time 长效长效Uptake by RES 被动到达肝脏、脾脏和骨髓被动到达肝脏、脾脏和骨髓Ligand-modified nanoparticle 可主动靶向分布于其他器官可主动靶向分布于其他器官Enhanced permeability 改变药物对生物膜的透过性，有利于改变药物对生物膜的透过性，有利于药物的胞内靶向传输药物的胞内靶向传输白蛋白纳米粒白蛋白纳米粒Hydrophobic
21、drugs,e.g.,Paclitaxel,docetaxel,rapamycin etc.AlbuminMean size=50-150 nmcryo-TEMConcentration dependent dissociation into individual drug-bound albumin moleculesActive drug in nanoparticle is in non-crystalline,amorphous,readily bioavailable stateTaxolAbraxane(nab-paclitaxel)is a solvent-free nano v
22、ersion of Taxol(cremophor-based paclitaxel)Contents:Paclitaxel 6 mg/mlCremophor 537 mg/mlEthanol 396 mg/mlContents:100 mg paclitaxel900 mg albuminNo Surfactants/SolventsAbraxane received FDA Approval January,2005for metastatic breast cancerAbraxaneNanomaterials Nanomaterials 纳米粒的载体材料纳米粒的载体材料Biodegra
23、dable 生物可降解生物可降解Biocompatible 生物相容性生物相容性Low toxicity 天然高分子材料天然高分子材料半合成高分子材料半合成高分子材料合成高分子材料合成高分子材料壳聚糖壳聚糖 Chitoson一种天然的聚阳离子多糖一种天然的聚阳离子多糖具有优良的生物相容性和生物降解性具有优良的生物相容性和生物降解性已经广泛的应用于医疗领域已经广泛的应用于医疗领域甲壳类动物甲壳类动物 甲壳素甲壳素 壳聚糖壳聚糖34聚乳酸聚乳酸聚乳酸聚乳酸-乙醇酸乙醇酸PLA PLGA乳酸乳酸丙交酯丙交酯乙醇酸乙醇酸乙交酯乙交酯36Polymerization method 聚合法聚合法 Disp
24、ersion method 分散法分散法 NP preparationNP preparation：物理分散法：物理分散法/化学反应法化学反应法micelle polymerizationemulsion polymerizationinterfacial polymerization emulsion dispersion salting-out dispersionsolvent evaporation SonicationPreparation of NanoparticlesEvaporationEvaporation乳化乳化-溶溶剂剂蒸蒸发发法法38Result39Appearanc
25、e of INS-PLC-NPs before and after lyophilization冻干前冻干前冻干品冻干品冻干后冻干后Characterization of INS-PLC-NPs before and after lyophilization40Therapeutic effect in diabetic rats after s.c.injection(n=5)3 Days41界面缩聚法 Interfacial polymerization 如氰基丙烯酸酯的单体在OH-离子的作用下，发生聚合反应，可生成聚氰基丙烯酸酯纳米粒。米托蒽醌聚氰基丙烯酸正丁酯毫微粒米托蒽醌聚氰基丙烯酸
26、正丁酯毫微粒 nanoparticle drug molecule NP loaded with drug Mitoxantrone polybutyl cyanoacrylate nanoparticles图图 米托蒽醌毫微粒的扫描电镜照片米托蒽醌毫微粒的扫描电镜照片 Tumor inhibition effects Tumor inhibition effectsOverall survival of HCC patients treated with DHAD-PBCA-NP or DHAD injection 治疗治疗108108例原发性肝癌效果（华西医院、四川省例原发性肝癌效果（华西
27、医院、四川省肿瘤医院、重庆市肿瘤医院、广西医科大学肿瘤医院）肿瘤医院、重庆市肿瘤医院、广西医科大学肿瘤医院）In vivoIn vivo distribution of NP distribution of NPYNanoparticles will usually be taken up by the liver,spleen and other parts of the RES depending on their surface characteristicsYNanoparticles with more hydrophobic surfaces will preferentially
28、 be taken up by the liver,followed by the spleen and lungsYParticles with longer circulation times,and hence greater ability to target to the site of interest,should be 100 nm or less in diameter and have a hydrophilic surface in order to reduce clearance by macrophages.Microemulsion Microemulsion 微
29、乳微乳Microemulsions are clear,stable,isotropic liquid mixtures of oil,water and surfactant,frequently in combination with a co-surfactant.由适当比例的表面活性剂，助表面活性剂，水和由适当比例的表面活性剂，助表面活性剂，水和油自发形成的各向同性、外观透明或半透明、热油自发形成的各向同性、外观透明或半透明、热力学稳定的分散体系力学稳定的分散体系 In contrast to ordinary emulsions,microemulsions form upon si
30、mple mixing of the components and do not require the high shear conditions generally used in the formation of ordinary emulsions.The two basic types of microemulsions are direct(oil dispersed in water,o/w)and reversed(water dispersed in oil,w/o).MicroemulsionEmulsionThermodynamically Stable Kinetica
31、lly StableDroplet size 10-100 nm(transparent)1-10 m(opaque)High surface area:200 m2/gLow surface area:15 m2/gW/O,O/W and Bicontinuous types W/O or O/W typesDifference between Microemulsions and Difference between Microemulsions and emulsionsemulsions50Advantages of microemulsion based Advantages of
32、microemulsion based drug deliverydrug deliveryspontaneous formation 自发形成，不需外界提供能量自发形成，不需外界提供能量ease of manufacturing and scale-upthermodynamic stability 热力学稳定，长期放置或离心不分热力学稳定，长期放置或离心不分层层 improved drug solubilization and bioavailability 提高难溶提高难溶性药物的溶解度，促进药物吸收，提高其生物利用度性药物的溶解度，促进药物吸收，提高其生物利用度 OilFormulat
33、ing MicroemulsionsFormulating MicroemulsionsCo-surfactantWaterO/WmicroemulsionW/Omicroemulsion52Method of microemulsion preparationMicrofluidization加水滴定法加水滴定法加油滴定法加油滴定法加乳化剂滴定法加乳化剂滴定法交替加入法交替加入法微乳的形成不需要外界做功，是体系内各组分达微乳的形成不需要外界做功，是体系内各组分达到适当的配比时自发形成的。到适当的配比时自发形成的。YLiposomeYPolymeric nanoparticles/macrop
34、articlesYBlock copolymer micellesYEmulsionYDendrimersClasses of particulate drug carriersPassive targeted drug delivery 被动靶向被动靶向Active targeted drug delivery 主动靶向主动靶向Physicochemical targeted drug delivery 物理化学靶向物理化学靶向Classification of TDDSClassification of TDDSPassive targeted drug delivery Passive
35、targeting relies on the natural distribution pattern of the drug or drug-carrier systemParticle size,Surface characters 粒径大小粒径大小 表面特征表面特征Less than 7m:removed from the blood by macrophages 巨噬细胞巨噬细胞 of the RES 网状内皮系统网状内皮系统 when administered systemicallyLarger than 7m:target drug to lungNanoparticles l
36、ess than 50 nm:accumulate in bone marrow 骨髓骨髓 slowly广泛应用于肿瘤靶向治疗中的一种靶向机制：广泛应用于肿瘤靶向治疗中的一种靶向机制：EPR effect(enhanced permeability and retention)增强的血管通透性和存留量增强的血管通透性和存留量 increased permeability of endothelial barriers in tumor blood vessels;the lack of effective lymphatic drainage from the tumor.血管壁间隙较宽，结血
37、管壁间隙较宽，结构完整性较差，淋巴构完整性较差，淋巴回流缺失回流缺失实体瘤组织实体瘤组织血管丰富血管丰富EPREPR效应（大分子物质和微粒在肿瘤效应（大分子物质和微粒在肿瘤组织具有的高通透性和滞留效应）组织具有的高通透性和滞留效应）Smooth muscle layer内皮间隙致密、内皮间隙致密、结构完整结构完整 被动靶向被动靶向Passive targeted drug delivery 被动靶向被动靶向Active targeted drug delivery 主动靶向主动靶向Physicochemical targeted drug delivery 物理化学靶向物理化学靶向Classi
38、fication of TDDSClassification of TDDSActive targeted drug delivery Active targeting employs a deliberately modified drug or drug carrier molecule capable of recognizing and interacting with a specific cell tissue or organ in the body 经特殊分子经特殊分子修饰的药物或药物载体，可将药物定向修饰的药物或药物载体，可将药物定向地运送到靶区浓集发挥药效。地运送到靶区浓集
39、发挥药效。Modification of the carrier system may include incorporation of antigen-specific antibodies 抗体抗体,or attachment of cell receptor-specific ligands 配体配体Active particulate Drug CarriersMechanism for active target delivery of drug-loaded particulates On binding to the receptors,the ligandparticle co
40、mplex could be internalized into the cell.Targeting ligand Target areaCell internalization mechanismFolic acidTumor cellCaveolin-assistedLow-density lipoporteinTumor cell,RES(liver,spleen)Caveolin-assistedMannose-6-phosphateRES(liver,lung,spleen),Dendritic cellClathrin-dependent Receptor-mediated en
41、docytosisTransferrin Tumor cell,brainArg-Gly-Asp(RGD)Integrins,Tumor cell,autoimmune diseasesCell adhesion molecule mediated endocytosisMonoclonal antibodyVarious targetGrowth factor familyTumor cell,cardiovascular disease，bone，nerve,etc.Major targeting ligands for drug delivery根据肝细胞表面过量表达去唾液酸糖蛋白受体，
42、根据肝细胞表面过量表达去唾液酸糖蛋白受体，能够特异性地结合半乳糖残基，对胆固醇能够特异性地结合半乳糖残基，对胆固醇分子进行半乳糖修饰。分子进行半乳糖修饰。半乳糖修饰的胆固醇脂质体半乳糖修饰的胆固醇脂质体不同半乳糖密度的胆固醇衍生物不同半乳糖密度的胆固醇衍生物不同链长的胆固醇衍生物不同链长的胆固醇衍生物Xun Sun,et al.J of Drug Targeting.2005,13:121-128.J of Drug Targeting.2010,18:520-535 A549 cellsHepG2 cells配体分子结构对配体分子结构对Gal-LPD转染率的影响转染率的影响Targeting
43、 MoietyTargeting MoietyAntigen binding fragments 66ProdrugProdrugDefinition:A pharmacologically inactive chemical entity that when metabolized or chemically transformed by a mammalian system is converted into a pharmacologically active substance.母体药物经化学衍生而成的物质，在体内经酶或母体药物经化学衍生而成的物质，在体内经酶或化学反应再生成母体药物，
44、发挥治疗作用化学反应再生成母体药物，发挥治疗作用 Improve patient acceptability(decrease pain on injection)Alter and improve absorption Alter biodistribution Alter metabolism Alter eliminationWhy use prodrugs?Why use prodrugs?Metabolism(enzyme dependant)Chemical Methods(non-dependant):Hydrolysis 水解水解 Decarboxylation 脱羧脱羧 NO
45、T patient dependant!Stability/Storage issuesConversion of ProdrugsConversion of ProdrugsPrincipals for the design of prodrugProdrug should be:Inactive and nontoxic Easily synthesizable Chemically stable outside site of action Bioreversible(parent drug must be regenerated in vivo)Target mechanism of
46、prodrugTarget mechanism of prodrugOCH3OOOOOHCH3CH3OCH3CH3CH3OHCH3CH3OHOOCH3OHCH3OMeCH3ONH(CH3)2+-SO4Erthromycin estolateIlosone-Eli Lillycaps,tabs,suspensionAntibiotic used to treatupper and lower respiratoryinfections(URI or LRI),Legionnaires disease,skin infections Erythromycin红霉素红霉素 is a very bit
47、ter substance easily destroyed at acidic pH Propionate ester is to increase lipid solubility for improved absorption Ester must be hydrolyzed for antibacterial activity Lauryl sulfate salt十二烷基硫酸盐十二烷基硫酸盐 absorption not affected by food,less bitter after taste and is acid stableFunctional Groups in Pr
48、odrugsAzo Prodrugs 偶氮前体药物偶氮前体药物Bacterial reductases reductive cleavage Occurs in colon discourages small intestine systemic absorption Concentrates the drug at the desired site of actionNNHSO2N NOHCO2HSulfasalazine-Azulfidine-Pharmacia&UpjohnSulfonamide antibiotic and antiinflammatoryUsed to treat U
49、lcerative colitis,rheumatoid arthritisNNHSO2NH2NH2OHCO2H+5-aminosalicylic acidSulfapyridine柳氮磺胺吡啶柳氮磺胺吡啶 73Key Lab of Drug Targeting and drug delivery systems,Sichuan University靶向药物及释药系统教育部重点实验室靶向药物及释药系统教育部重点实验室四川大学华西药学院四川大学华西药学院14-Succinate lysozyme conjugate for the renal delivery of triptolide(雷公藤
50、）Renal disease and systemic disease caused by renal disease are frequently occurred and difficult to treat all over the world.Its reported that the incidence of kidney disease was up to 9.6%in China,and more than 90%of patients know very little about their disease.The incidence of End Stage Renal Di
