温度及pH值敏感水凝胶的合成和应用

直接将丙烯酸单体与N-异丙基丙烯酸胺共聚交联合成了温度及pH值敏感的水凝胶。包埋于水凝胶中的药物的释放随温度升高和pH值增大而加快，药物的释放兼有温度和pH值敏感性，对pH值的响应更加显著。     

可生物降解肝素钠/两性壳聚糖复合物用于蛋白药物pH响应释放研究

制备了一类可生物降解肝素钠两性壳聚糖复合物(HPACS),并探索将其用于蛋白药物pH响应释放.两性壳聚糖由壳聚糖与丙烯酸加成反应得到,丙烯酸取代度可通过丙烯酸壳聚糖投料比调控;用胶体与pH浊度滴定研究了肝素钠与两性壳聚糖的复合作用,发现两组分在一定pH范围内能通过静电相互作用形成复合物,复合转变临界pH(pHΦ)与两性壳聚糖中丙烯酸取代度有关,取代度越低,pHΦ值越高.以牛血清白蛋白(BSA)为模型,测定了其在复合物中包埋及不同pH介质中的释药行为.结果表明,BSA可以在非常温和条件下有效包埋于复合物中,包埋率接近100%;BSA从复合物中释放具有很高的pH响应性,释放转变在很窄的pH范围内(<0.4pH单位)完成,释放转变临界pH(pH′Φ)可由两性壳聚糖中丙烯酸取代度调控.复合物形成和蛋白质释放在对pH依赖性上存在很好的相关性.同时还发现,在中性介质中(pH7.4),复合物对BSA具有很好的缓释作用,BSA持续释放时间可达15天左右.     

具有pH-响应性的聚天冬氨酸类载药胶束的制备和释放能力研究

通过大分子引发剂ω-氨基-α-甲氧基聚乙二醇引发N-羧基-α-氨基环内酸酐开环聚合和水合肼侧基改性,制备了一系列聚乙二醇-聚氨基酸类三嵌段共聚物.其中聚氨基酸链段包括具有酰肼基的聚天冬氨酸衍生物(PAHy),以及疏水性的聚丙氨酸链段.引入具有pH响应性的腙键键合阿霉素,利用键合阿霉素与游离阿霉素之间的π-π叠合作用,在聚合物自组装形成胶束过程中通过化学键合+物理包埋的方式充分负载药物.该胶束以聚丙氨酸链段为核心,以PEG链段为冠层,以PAHy链段为包裹药物的壳层.载药胶束的粒径在170 nm左右.研究不同pH值条件下载药胶束的药物释放能力,随环境pH值的降低药物的释放速率显著增加.     

壳聚糖-g-聚甲基丙烯酸凝胶粒的制备及其药物释放行为

以壳聚糖和甲基丙烯酸为原料,硝酸铈铵为引发剂,合成了不同接枝率的壳聚糖-g-聚甲基丙烯酸(CS-g-PMAA),用FTIR、1H NMR和元素分析表征了产物的结构,以柠檬酸三钠和戊二醛为交联剂制备了具有核壳结构的CS-g-PMAA载药体系。 用UV/Vis检测了CS-g-PMAA粒子对模型药物的释放行为。 结果表明,CS-g-PMAA接枝率为12.21%时药物释放速率最慢,其在pH=1.8介质中药物累积释放量(11 h)为44.18%,而壳聚糖粒子的累积释放量高达65.24%,即接枝改性壳聚糖粒子对药物的缓慢控制释放性能较好; CS-g-PMAA粒子的释药行为还依赖于介质的pH值和盐浓度,在低pH值和低盐浓度下,药物释放速率较快;酶环境下由于载体材料的降解使药物释放速率加快。 分析了不同条件下CS-g-PMAA载药粒子中药物的释放机理。     

逐层自组装微囊对甲氨蝶呤的体外可控释放研究

高装载率和可控释放的药物传送系统具有很好的生物医学应用前景.本文研究了逐层自组装技术对抗肿瘤药物甲氨蝶呤(MTX)的装载与控释.利用不同颜色的发光量子点将甲氨蝶呤与高分子聚电解质进行标记后,通过添加Tween 80一锅法制备出平均粒径约500 nm,装载率为54.82%的MTX碳酸钙颗粒,并以此为模板制备出了可酸度控释的MTX药物微囊.探讨了酸度、逐层自组装层数、药物分子的标记等因素与可控释放的密切关系及相关机理.此项研究可为MTX药物微囊的细胞内实验及生物体内的应用奠定实验基础.     

靶向纳米钻石-甲氨蝶呤药物体系制备及与MCF-7细胞作用

以羧基化纳米钻石(ND-COOH)为基体, 通过共价键合方法将聚乙二醇二胺(H₂N-PEG-NH₂)、 叶酸(FA)和缩水甘油(GLY)偶联于ND-COOH表面, 赋予纳米钻石载体较好的水溶分散性和靶向性, 借助氢键和范德华力等作用力负载甲氨蝶呤(MTX), 得到靶向纳米钻石-聚乙二醇二胺-叶酸/缩水甘油/甲氨蝶呤(ND-PEG-FA/GLY/MTX NPF/G/M)纳米药物体系. 采用透射电子显微镜、 X射线能量色散谱、 粒径及电位测试证实已制备NPF/G/M. 体外释药发现NPF/G/M在肿瘤环境(pH=5.5)中的药物释放量为正常生理环境(pH=7.4)中的3倍, 表明其具有良好的药物输送特性. 此外, 利用流式细胞术和MTT毒性测试探究了MCF-7细胞摄取NPF/G/M的机制及动力学特性和细胞毒性, 结果表明NPF/G/M以依赖能量、 温度、 网格蛋白、 小窝蛋白和叶酸受体介导的机制进入细胞, 从而将药物缓慢释放于细胞内, 进而诱导细胞凋亡. 研究结果表明, NPF/G/M可作为一种良好的药物输送体系, 为其应用于乳腺癌的临床治疗提供理论参考.     

基于壳聚糖的pH响应双重药物释放系统研究

以腙键连接的壳聚糖阿霉素前药偶联物(Chitosan-hz-DOX)为载体,通过物理包埋法制备了负载喜树碱(CPT)的双药共传递纳米输送体系(CPT-CS-DOX).通过紫外可见吸收光谱、动态光散射、透射扫描电镜等方法研究了体系的粒径、形貌、药物负载及释放性能,发现制备CPT-CS-DOX纳米颗粒的最佳CPT投放量为20%,其粒径随着Chitosan-hz-DOX中阿霉素(DOX)含量的增加而不断降低,共传递体系有效地抑制了DOX和CPT的早期泄露,并呈现出显著的p H依赖药物释放行为.利用Peppas方程对释放曲线进行分析,发现第一阶段DOX和CPT在中性环境中的释放遵循Fick扩散控制和溶胀控制机理,在酸性环境中CPT的释放机理保持不变,而DOX的释放则转变为聚合物松弛机理;第二阶段则两者均符合Fick扩散机理.     

羧甲基淀粉微球的制备及其对阿司匹林的控制释放

以马铃薯淀粉为原材料,水-乙醇为溶剂制备了羧甲基淀粉(CMS)。通过反相微乳液法包埋阿司匹林制备了载药羧甲基淀粉微球。通过热重、X-衍射以及扫描电镜对羧甲基淀粉进行了表征。结果显示,淀粉经过羧甲基改性后结晶度降低,聚集形态变得更加不规则,说明淀粉羧甲基后在水相中溶解度增大,有利于制备淀粉微球。将羧甲基淀粉与阿司匹林溶于水中,配成浓度为8%的水相溶液。以环己烷与氯仿的混合溶剂(V_(环己烷):V_(氯仿)=3:1)作为油相体系,经过反相乳化成球,实现了对阿司匹林的包埋。羧甲基淀粉微球经载药后呈圆形,表面光滑,粒径为200~800nm,且实现了在不同p H值溶液中对小分子药物的控制释放。有望用于口服药的载体,实现在体内的控制释放,从而增加药物利用率以及降低药物的副作用。     

差别性释放两种药物的层层组装聚合物膜

基于层层组装技术制备了能够差别性释放2种药物的聚合物复合膜.通过大分子前体药物透明质酸-阿霉素(HA-DOX)与壳聚糖(HACC)的层层组装以及膜内后扩散负载甲氨喋呤二钠盐(MTX)的方法,实现了DOX和MTX 2种药物分子在聚合物膜中的负载.DOX和MTX在癌变组织的酸性环境下具有差别性的释放动力学,MTX在24 h内快速释放,而DOX长效缓释达10 d.细胞实验结果表明,差别性释放的DOX和MTX可有效地抑制癌细胞的增殖.     

pH调控Y型分子筛负载阿霉素纳米药物制备及与MM-231细胞的作用

针对抗肿瘤小分子药物靶向性差、疗效低和毒副性大等缺陷,我们以Y型分子筛(YMS)为基体、阿霉素(DOX)为药物模型,通过pH调控,借助氢键和范德华力等物理作用力制备得到高负载Y型分子筛纳米药物体系(YMS?DOX)。采用UV?Vis、FT?IR、粒径和电位测试及荧光光谱证实YMS?DOX成功制备,且DOX的负载率可高达99.61%。体外药物释放测试发现YMS?DOX具有pH响应释放特性,在肿瘤环境中(pH=4.5)的药物释放量为正常生理环境(pH=7.4)中的3.8倍,表明其具有良好的药物输送特性。此外,利用流式细胞术和MTT测试法探究了YMS?DOX对乳腺癌细胞(MM?231)和树突细胞(DC)的细胞凋亡和毒性,结果表明YMS?DOX可以诱导肿瘤细胞凋亡,且可降低对正常细胞的毒副作用。     

pH敏感型mPEG-Hz-PLA聚合物纳米载药胶束的制备

以合成的含有腙键的聚乙二醇大分子(mPEG-Hz-OH)为引发剂,以丙交酯为单体引发开环聚合反应,并通过调整投料比,制备出3种不同分子量的含腙键的生物可降解嵌段聚合物(mPEG-Hz-PLA).将腙键引入到聚合物的骨架中,以此构建聚合物胶束并作为pH敏感型纳米药物载体.制备的pH敏感型胶束的CMC值等于或低于5.46×10-4 mg/m L,DLS和TEM显示粒径均小于100 nm,且粒径分布均匀.非pH敏感型胶束在不同pH下的粒径变化不明显,而pH敏感型胶束在酸性环境下(pH=4.0和pH=5.0)胶束粒径出现了明显变化.以阿霉素为模型药物制备了pH敏感型载药胶束,其粒径比空白胶束大(100~200 nm),且粒径分布均匀.药物释放实验表明pH敏感型载药胶束随着释放介质pH降低累积释药量增高.MTT实验表明空白胶束对HeLa细胞和RAW264.7细胞几乎没有抑制作用,而载阿霉素的胶束对2种细胞的抑制作用都随着剂量的增大和时间的延长而增强.     

Co-delivery of docetaxel and doxorubicin using biodegradable PEG-PLA micelles for treatment of breast cancer with synergistic anti-tumour effects

Poly(ethylene glycol)-poly(lactic acid) copolymer, prepared by ring opening polymerization, was used as a single platform to co-deliver both hydrophilic doxorubicin and hydrophobic docetaxel (DTX) in a simulated physiological environment. The average size of the negatively charged drug loaded polymeric micelles were found to be 293 nm. The drug loading (%) and encapsulation efficiency (%) were calculated to be 1.21 and 59.0, respectively. The in vitro cytotoxicity test using MCF7 breast cancer cells was conducted using 1 × 10⁴ cells in 10% FBS and 1% antibiotic, and the absorbance of formazan was evaluated at 570 nm. Cell growth inhibition by MTT assay showed viability of 33% of the MCF7 cells after treatment with drug-loaded micelles for 48 h. Controlled release of drugs from the polymeric micelles indicated a burst release effect initially; whereas, 98% of drug could be released at pH 7.4 within a time period of 96 h. Time period for drug release shorten to 48 h only in simulated mild acidic pH (5.4) condition. The in vitro drug release study from micelles indicated synergistic cytotoxicity effect in human metastatic breast cancer MCF7 cell.     

Drug release and biocompatibility of self‐assembled micelles prepared from poly (ɛ‐caprolactone/glycolide)‐poly (ethylene glycol) block copolymers

A series of poly(?‐caprolactone/glycolide)‐poly(ethylene glycol) (P(CL/GA)‐PEG) diblock copolymers were prepared by ring opening polymerization of a mixture of ?‐caprolactone and glycolide using mPEG as macro‐initiator and stannous octoate as catalyst. Self‐assembled micelles were prepared from the copolymers using nanoprecipitation method. The micelles were spherical in shape. The micelle size was larger for copolymers with longer PEG blocks. In contrast, the critical micelle concentration of copolymers increased with decreasing the overall hydrophobic block length. Drug loading and drug release studies were performed under in vitro conditions, using paclitaxel as a hydrophobic model drug. Higher drug loading was obtained for micelles with longer poly(ε‐caprolactone) blocks. Faster drug release was obtained for micelles of mPEG2000 initiated copolymers than those of mPEG5000 initiated ones. Higher GA content in the copolymers led to faster drug release. Moreover, drug release rate was enhanced in the presence of lipase from Pseudomonas sp., indicating that drug release is facilitated by copolymer degradation. The biocompatibility of copolymers was evaluated from hemolysis, dynamic clotting time, and plasma recalcification time tests, as well as MTT assay and agar diffusion test. Data showed that copolymer micelles present outstanding hemocompatibility and cytocompatibility, thus suggesting that P(CL/GA)‐PEG micelles are promising for prolonged release of hydrophobic drugs.     

Thermo and pH Dual-Responsive Micelles of N-phthaloylchitosan-g-Poly(N-isopropylacrylamide) and Poly(acrylic acid-co-tert-butyl acrylate) for Drug Delivery

A novel amphiphilic copolymer N-phthaloylchitosan graft poly(N-isopropylacrylamide) and poly(acrylic acid-co-tert-butyl acrylate) (PHCS-g-PNIPAAm&P(AA-co-tBA)) was synthesized. The graft copolymer could form micelles in aqueous medium, and the critical micelle concentration (CMC) of the copolymer was 7.5 × 10^{? 3}mg/mL. The lower critical solution temperature (LCST) of the micelles was measured to be 30°C. Transmission electron microscopy (TEM) image showed that the micelles exhibited a regular spherical shape, and the mean diameter of the micelles was 94.1 ± 0.8 nm as determined by dynamic light scattering (DLS). The potential usefulness of the micelles as drug delivery systems was investigated using anti-inflammation drug prednisone acetate as the model. The drug loading capacity of the micelles was measured to be 22.86 wt%, and the DLS results showed that the mean diameter of the drug-loaded micelles was 133.3 ± 2.4 nm. In vitro drug release studies indicated that the micelles exhibited thermo and pH dual-responsive release profiles.     

Synthesis of self-assemble pH-responsive cyclodextrin block copolymer for sustained anticancer drug delivery

Well-defined p H-responsive poly(ε-caprolactone)-graft-β-cyclodextrin-graft-poly(2-(dimethylamino)ethylmethacrylate)-co-poly(ethylene glycol) methacrylate amphiphilic copolymers(PCL-g-β-CD-g-P(DMAEMA-co-PEGMA)) were synthesized using a combination of atom transfer radical polymerization(ATRP),ring opening polymerization(ROP) and "click" chemistry.Successful synthesis of polymers was confirmed by Fourier transform infrared spectroscopy(FTIR),proton nuclear magnetic resonance(1H-NMR),and gel permeation chromatography(GPC).Then,the polymers could selfassemble into micelles in aqueous solution,which was demonstrated by dynamic light scattering(DLS) and transmission electron microscopy(TEM).The p H-responsive self-assembly behavior of these copolymers in water was investigated at different p H values of 7.4 and 5.0 for controlled doxorubicin(DOX) release,and these results revealed that the release rate of DOX could be effectively controlled by altering the p H,and the release of drug loading efficiency(DLE) was up to 88%(W/W).CCK-8 assays showed that the copolymers had low toxicity and possessed good biodegradability and biocompatibility,whereas the DOX-loaded micelles remained with high cytotoxicity for He La cells.Moreover,confocal laser scanning microscopy(CLSM) images revealed that polymeric micelles could actively target the tumor site and the efficient intracellular DOX release from polymeric micelles toward the tumor cells further confirmed the anti-tumor effect.The DOX-loaded micelles could easily enter the cells and produce the desired pharmacological action and minimize the side effect of free DOX.These results successfully indicated that p H-responsive polymeric micelles could be potential hydrophobic drug delivery carriers for cancer targeting therapy with sustained release.     

核交联的含磺酸甜菜碱pH敏感胶束的制备与表征

制备了一种在疏水段带有侧基叠氮官能团的两亲性pH敏感的聚合物——聚己内酯-聚(甲基丙烯酸二乙氨基乙酯-磺酸甜菜碱)((PCL-ACL)-PDEAS);同时合成了两端带有炔基中间带有二硫键的交联剂,用红外、核磁表征了目标分子.通过两亲性高分子自组装形成胶束,并通过点击化学反应获得了核交联的胶束.通过动态光散射测定粒径,胶束酸碱滴定表征胶束的pH敏感性,还原条件下释放药物的速度,对比了非交联胶束和交联胶束的性质.结果表明,交联胶束在正常生理条件下的释放速度比未交联胶束更慢;而在有DTT的存在条件下,交联胶束由于二硫键断裂,释放速率明显加快.因此,核交联载药胶束有可能响应肿瘤的微环境实现靶向释放.     

Natural Triterpenoid‐ and Oligo(Ethylene Glycol)‐Pendant‐Containing Block and Random Copolymers: Aggregation and pH‐Controlled Release

In this research, a series of random and block amphiphilic copolymers of norbornene derivatives containing biocompatible natural triterpenoid and oligo(ethylene glycol) pendants were synthesized by ring‐opening metathesis polymerization. These copolymers were heat and pH responsive, and could self‐assemble into core–shell spherical micelles in aqueous solution. Their hydrodynamic diameters corresponded to pH values and monomer sequences. By evaluating the loading and release capacity of hydrophobic molecules, it was found that 1) the higher the content of the hydrophobic triterpenoid, the higher the loading capacity; 2) the release speed could be trigged by the pH because of the deprotonation of the carboxyl groups on the triterpenoid. Additionally, the copolymers exhibited low cytotoxicity toward L929 cells, which makes them potential nanocarrier candidates for controlled drug delivery.     

Synthesis,self-assembly,and pH-responsive drug release of PHMEMA-PEG-PHMEMA ABA triblock copolymers

Abstract

A series of tertiary amine containing PHMEMA-PEG-PHMEMA ABA triblock copolymers were synthesized by atom transfer radical polymerization (ATRP) using bromine-capped poly(ethylene glycol) (Br-PEG-Br) and 2-(hexamethyleneimino)ethyl methacrylate (HMEMA) as macro-initiator and monomers, respectively. The chemical structures and molecular weights of triblock copolymers were characterized by ¹H NMR and gel permeation chromatography (GPC). The self-assembly behaviors of copolymers in different pH conditions were studied by dynamic light scattering (DLS) and transmission electron microscopy (TEM). Triblock copolymers self-assembled into micelles in water (pH 7.4) and the micelles disassembled at acidic pH (pH 5.0). Anticancer drug doxorubicin (DOX) was used as a drug model and physically encapsulated into polymeric micelles. The drug release of DOX-loaded polymeric micelles was pH-responsive; the drug-loaded micelles that had higher contents of tertiary amine in polymer pendant groups showed faster release speed. In addition, the drug-loaded micelles showed excellent inhibition efficacy against HeLa cells in vitro.     

荧光标记pH敏感胶束的制备及其药物控释

利用原子转移自由基聚合方法（ATRP）合成了pH敏感的两亲性嵌段共聚物mPEG-b-PDPA_n（聚合度n=100-200）及荧光修饰的嵌段聚合物异硫氰酸荧光素-聚乙二醇-聚N,N-二异丙胺基甲基丙烯酸乙酯（FITCPEG₄₅-PDPA₁₀₀）。采用溶剂挥发的方法制备胶束,此胶束呈现均一的球形分布,平均粒径180-240 nm（0.3 mg·mL^-1）。以阿霉素（DOX）为模拟药物,其胶束载药量约11%（w,质量分数）左右,外环境pH对载药胶束的粒径和体外释放行为有显著影响。在弱酸环境下,胶束核质子化发生膨胀甚至解体,在2-3 h内药物可释放80%左右。体外毒性试验表明,空白胶束与人类肝癌细胞（Huh7）有良好的生物相容性。同时,与此细胞共同孵育5 h的荧光聚合物胶束体现了较好的转染效果。因此,这类荧光标记胶束可能会为实时跟踪化疗药物的输送或分布打开新的视角。     

Preparation and characteristics of linoleic acid-grafted chitosan oligosaccharide micelles as a carrier for doxorubicin

The linoleic acid (LA)-grafted chitosan oligosaccharide (CSO) (CSO-LA) was synthesized in the presence of 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide (EDC), and the effects of molecular weight of CSO and the charged amount of LA on the physicochemical properties of CSO-LA were investigated, such as CMC, graft ratio, size, zeta potential. The results showed that these chitosan derivatives were able to self-assemble and form spherical shape polymeric micelles with the size range of 150.7–213.9 nm and the zeta potential range of 57.9–79.9 mV, depending on molecular weight of CSO and the charged amount of LA. Using doxorubicin (DOX) as a model drug, the DOX-loaded CSO-LA micelles were prepared by dialysis method. The drug encapsulation efficiencies (EE) of DOX-loaded CSO-LA micelles were as high as about 75%. The sizes of DOX-loaded CSO-LA micelles with 20% charged DOX (relating the mass of CSO-LA) were near 200 nm, and the drug loading (DL) capacity could reach up to 15%. The in vitro release studies indicated that the drug release from the DOX-loaded CSO-LA micelles was reduced with increasing the graft ratio of CSO-LA, due to the enhanced hydrophobic interaction between hydrophobic drug and hydrophobic segments of CSO-LA. Moreover, the drug release rate from CSO-LA micelles was faster with the drug loading. These data suggested the possible utilization of the amphiphilic micellar chitosan derivatives as carriers for hydrophobic drugs for improving their delivery and release properties.