含RGD修饰的丹参酮ⅡA多级靶向纳米粒的质量评价

目的探讨丹参酮ⅡA多级靶向纳米粒的制备及其质量评价。方法采用乳化溶剂蒸发法制备丹参酮ⅡA多级纳米粒;测定丹参酮ⅡA多级靶向纳米粒的粒径分布及纳米微粒表面结构;并检测丹参酮ⅡA多级靶向纳米粒的载药量、包封率及体外药物释放规律。结果课题组制备的丹参酮ⅡA多级靶向纳米粒,大小均匀,载药纳米粒的平均粒径为190nm,Zeta电位为4.3mV,包封率(94.12±5.20)%,载药量(2.05±0.12)%。与游离的丹参酮ⅡA单体相比,丹参酮ⅡA多级靶向纳米粒释放速度明显减慢,在120h累积释放量为72.59%。结论采用乳化溶剂蒸发法成功制备了含RGD修饰的丹参酮ⅡA多级靶向纳米粒。与丹参酮ⅡA单体相比,制备成纳米制剂后,多级靶向载药纳米粒能随着时间的延长将药物逐步释放出来,具有良好的缓释特征。     

PEG-胆固醇双重修饰PBCA纳米粒脑靶向机制

目的中枢神经系统疾病目前存在的主要问题是血脑屏障(BBB)通透性的问题,本实验期望通过制备PEG-胆固醇双重修饰PBCA纳米粒,提高难透过BBB的中枢神经系统疾病的治疗及早期诊断药物的治疗作用,并对其经过BBB进入脑组织的机制进行探讨。方法采用乳化聚合法制备双重修饰PBCA纳米粒并对其理化特性进行评价。(1)双重修饰PBCA纳米粒体内药动学及脑组织分布研究:实验前12小时大鼠进行颈静脉插管手术。实验时于插管处给予原药溶液,胆固醇单修饰及PEG-胆固醇双重修饰PBCA纳米粒。给药后不同时间点取血200μL,化学/发光分析仪测定血液中香豆素-6的含量。大鼠尾静脉分别给予原药溶液、胆固醇单修饰及PEG-胆固醇双重修饰PBCA纳米粒。给药后不同时间点处死大鼠,取脑组织测定香豆素-6的含量。(2)双重修饰PBCA纳米粒体外细胞摄取及转运机制研究:采用b End.3细胞模型(永生化小鼠脑内皮细胞株)评价了载体及纳米粒的体外细胞毒性,同时选用了巨胞饮介导的内吞作用(细胞松弛素D),网格蛋白介导的内吞作用(氯丙嗪),小窝蛋白介导的内吞作用(染料木素),胆固醇耗竭(MβCD和制霉菌素),高尔基体抑制剂(布雷菲德菌素A)和耗能(叠氮钠)等七种细胞摄取抑制剂来研究载药纳米粒的细胞摄取及转运机制。结果 (1)制备的纳米粒粒径为(191.1±1.22)nm、载药量约为1.97%,包封率大于98%。同时双重修饰PBCA纳米粒在体外具有更明显的缓释效果。(2)双重修饰PBCA纳米粒在体内具有更高的血药浓度及缓释效果,能持续释放12 h,而胆固醇单修饰纳米粒仅释放8 h,原药溶液仅4 h内能检测到药物。脑组织分布研究结果表明,双重修饰PBCA纳米粒静脉给药后脑组织中药物含量显著增高且成缓慢释放的过程,说明具有成为脑靶向制剂的研究潜力。(3)载体及载药纳米粒细胞安全性良好,纳米粒在实验范围内没有明显的细胞毒性,具有较好的安全性。选择不同类型的摄取和转运抑制剂研究了载药纳米粒的细胞摄取及转运机制。结果表明,参与细胞转运和摄取的机制可以是不同的,而胆固醇-PEG双修饰PBCA纳米粒在b End.3细胞中的转运机制,是一个包括巨胞饮介导、耗能、同时需要胆固醇参与等多种机制介导的复杂过程。结论本实验制备的PEG20000-胆固醇双重修饰PBCA纳米粒,可解决中枢神经系统疾病的治疗及诊断药物体内BBB通透率低、安全性差、缓释性差等瓶颈问题,具有重要应用前景。     

阿苯达唑聚氰基丙烯酸正丁酯纳米粒的制备、性质及其组织靶向性研究阿苯达唑聚氰基丙烯酸正丁酯纳米粒的制备、性质及其组织靶向性研究

目的制备阿苯达唑聚氰基丙烯酸正丁酯纳米粒（albendazole polybutycyanocrylate nanoparticles，ABZ-PBCA-NP）TDDS给药系统，并考察相关特性及组织分布靶向性。方法种子乳化聚合法制备阿苯达唑纳米粒；等温吸附法考察纳米粒载药特性；动态透析法研究4种制剂的体外释药动力学；同位素标记阿苯达唑纳米粒在小鼠脏器组织分布和生物利用度。结果ABZ-PBCA-NP体外释药遵循Higuchi方程，加入PVP制成的载药纳米粒符合双指数函数。纳米粒的载药方式遵循Langmuir吸附方程。小鼠ig ³H-ABZ-PBCA-NP后, 药物的肝、脾中的靶向指数分别为11.4和3.9，阿苯达唑纳米粒和混悬剂相对生物利用度分别为76.0%和36.9%。结论制备纳米粒加入PVP可使药物具吸附性和分散性，纳米粒载体可降低药物与血浆蛋白结合率，增强药物的肝、脾脏器靶向性和延缓释药。     

抗 EGFR 单克隆抗体偶联载紫杉醇纳米粒对胃癌肿瘤靶向性能力研究

目的：为了增强载药纳米粒靶向肿瘤的能力,将载药纳米粒与抗上皮生长因子细胞增殖和信号传导受体（EGFR）单克隆抗体偶联,探究该偶联是否会增加胃癌肿瘤靶向性。方法将制备好的载药纳米粒在羟基胶乳微球偶联抗体（EDC）的作用下通过酰胺反应与抗 EGFR 单克隆抗体进行偶联,制备成载药免疫纳米粒。通过透析袋法测定纳米粒体外缓释效果,用 ELISA 法测定抗体与载药纳米粒偶联率,最后采用荧光标记法对胃癌模型鼠进行活体成像结果纳米粒表面光滑,分布均匀,粒径在200～300 nm 间,载药量为61．02％,并且具有缓慢释药的效果,抗体与载药纳米粒的偶联率达到了70％,最终荧光活体成像的结果显示,此免疫载药抗体具有靶向胃肿瘤的效果。结论我们制备的免疫纳米粒既可以缓释药物也存在一定肿瘤靶向性,所以将抗体与载药纳米粒偶联,可以增加肿瘤病灶处局部的药物浓度,以此来缓解抗肿瘤药物对人体其他正常组织毒副作用。     

壳聚糖载药纳米粒研究进展

目的介绍壳聚糖载药纳米粒近年来的研究进展。方法总结壳聚糖纳米粒的制备方法、释药特性、生物摄取及其应用。结果不同的制备方法可得到不同粒径和表面特性的壳聚糖纳米粒。壳聚糖纳米粒改变了壳聚糖的摄取机制，广泛应用于药物的器官靶向、DNA转染效率提高、药物的非注射途释给药等方面。结论壳聚糖纳米粒作为一种新型的药物载体，具有重要的研究开发价值。     

叶酸壳寡糖修饰的紫杉醇PLGA纳米粒的体外释放及对SKOV-3增殖的抑制作用

摘 要 目的： 制备叶酸壳寡糖修饰的紫杉醇PLGA纳米粒（F-CS-PLGA-NPs）,并考察其对人卵巢癌上皮细胞（SKOV-3）的抑制作用。方法: 采用界面沉积法制备F-CS-PLGA-NPs,以30%乙醇作为释放介质考察修饰和未修饰纳米粒的体外释药情况,MTT法比较不同剂型和不同浓度紫杉醇对SKOV-3增殖的抑制作用。结果: F-CS-PLGA-NPs的粒径为（321±0.76）nm,电位为（22.6±0.26）mV,载药量为（5.1±0.25）%,包封率为（41.96±1.96）%。修饰和未修饰纳米粒的体外释药曲线相似,在最初24 h内约有35%药物释放,随后释药速度减慢,纳米粒以近零级方式释放,144 h累计释药率约为75%。细胞实验结果显示,在紫杉醇浓度相同的情况下,F-CS-PLGA-NPs在不同作用时间对细胞的抑制作用均强于紫杉醇溶液组和普通纳米粒组,F-CS-PLGA-NPs对SKOVS细胞增殖的抑制作用在一定程度上被游离叶酸减弱。结论:叶酸壳寡糖的修饰增加了纳米粒对SKOVS 3细胞的靶向性。     

无稳定剂修饰的汉防己甲素PLGA纳米粒制备及体外评价

目的:制备无稳定剂修饰的汉防己甲素PLGA纳米粒,研究其理化性质及细胞毒和细胞摄取特性。方法:以聚乳酸-羟基醋酸共聚物(PLGA)为载体材料,采用无稳定剂修饰的纳米沉淀法制备汉防己甲素纳米粒;通过单因素试验考察不同制备工艺对纳米粒理化性质的影响;通过载药量、包封率、累积释药量等指标考察其载药特性;采用MTT比色法检测其对人肺腺癌细胞株A549的细胞毒性;采用共聚焦显微镜技术考察其细胞摄取特性。结果:无稳定剂修饰的汉防己甲素PLGA纳米粒平均粒径169.3 nm,与有稳定剂的汉防己甲素PLGA纳米粒相比外观无明显改变。在一定范围内,随着PLGA用量的增加,纳米粒的粒径呈上升趋势;随着投药量的增加,纳米粒的载药量显著增加,包封率下降。在pH7.4的释放介质中,纳米粒释慢释药,96 h累积释药率60.44%。细胞毒试验显示,当培养时间为8 h时,汉防己甲素组的细胞毒性大于汉防己甲素纳米粒组;当培养时间延长至24 h时,汉防己甲素纳米粒组的细胞活性明显低于纯药物组;高剂量的空白纳米粒组始终表现较低的细胞毒性。激光共聚焦电镜断层扫描显示汉防己甲素纳米粒能够较好的被细胞摄取。结论:制备的无稳定剂修饰的汉防己甲素PLGA纳米粒大小均一,包封率高,体外释药表现出较好的缓释效果,易被细胞摄取,对A549细胞的增殖有明显的抑制作用。     

抗肿瘤药物靶向载体系统的研究与开发

综述肿瘤靶向给药的基础和抗肿瘤药物靶向载体系统的发展。分类介绍普通被动靶向载药系统（如微乳、传统脂质体、聚合物纳米粒、固体脂质纳米粒、纳米脂质载体、药-脂结合物纳米粒等）、表面修饰的被动靶向载药系统及主动靶向载药系统（如免疫脂质体、免疫聚合物纳米粒及受体-配体介导靶向纳米载体）的研究与开发。在传统药物制剂的基础上，发展抗肿瘤药物的新型靶向载体系统，改善药物在体内的代谢动力学特性，增加药物定向富集到肿瘤部位甚至肿瘤细胞内，提高疗效，降低毒副作用，是近年来备受关注的课题。     

载肿瘤坏死因子拮抗肽PEG-PLGA纳米粒与血清蛋白作用研究

目的:研究聚乙二醇(polyethyleneglycol,PEG)链长、含量以及载药对聚乙二醇-聚乳酸羟基乙酸[polyethyleneglycol modified poly(D,L-lactide-co-glycolide),PEG-PLGA]纳米粒吸附血清蛋白的影响,研究负载肿瘤坏死因子-α拈抗肽(tumor necrosis factor alpha blocking peptide,TNF-BP)的PEG-PLGA纳米粒在血清中药物释放行为.方法:采用二喹啉甲酸(bicinchoninic acid,BCA)蛋白定量试剂盒和十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(sodium dodecyl sulfate-polyacrylamide gel electrophoresis,SDS-PAGE)技术检测吸附于纳米粒表面的血清蛋白,采用紫外分光光度法检测载药PEG-PLGA纳米粒在血清中释放TNF-BP.结果:PEG链长以相对分子质量表示为5 000,含量为7.5%时,空载纳米粒吸附血清蛋白为(2.57±0.19)%;负载TNF-BP后纳米粒吸附血清蛋白能力降低.载药纳米粒在血清中存在突释效应,并随着PEG链长的增加,累积释放率增大.PEG含量由2.5%变化至10.0%时,对释药性能影响程度不明显.结论:负载TNF-BP和PEG修饰可降低血清蛋白在纳米粒表面的吸附,载药PEG-PLGA纳米粒在血清中的累积释药率大于PLGA纳米粒.     

HZ08新型仿生纳米粒的构建与性质研究

目的：研究具有肿瘤多药耐药逆转活性的四氢异喹啉类化合物HZ08-重组高密度脂蛋白（reconstituted high density lipoprotein,rHDL）纳米粒的构建及性质,旨在研制HZ08的新型仿生纳米制剂。方法：通过胆酸钠法制备HZ08-rHDL纳米粒,考察该纳米粒的包封率、载药量、渗漏率、形态、粒径等理化性质,透析法研究制剂的体外释放特性,MTT法考察rHDL载体对人乳腺癌细胞（MCF-7）和人正常乳腺细胞（MCF-10A）的毒性,HPLC法、流式细胞术和荧光显微术评价纳米粒的肿瘤细胞靶向性。结果：HZ08-rHDL纳米粒包封率（93.45±0.28）%,载药量（10.65±0.46）%,水分散液于4℃放置一个月渗漏率为（4.50±0.12）%,外观呈圆整球形,平均粒径为（105.53±2.50） nm;体外48 h 累积释放量仅为（12.13±1.08）%;rHDL 载体细胞毒性低,且MCF-7细胞对rHDL荷载药物的摄取能力显著强于MCF-10A细胞（P＜0.0001）,有较强的体外肿瘤靶向性。结论：HZ08-rHDL纳米粒的包封率高、性质稳定、粒径大小适宜、缓释效果明显、载体毒性低且有较好的肿瘤细胞靶向性,具有继续研究开发的价值。     

Superparamagnetic nanovector with anti-cancer properties: γFe2O3@Zoledronate

We elaborate a magnetic nanovector to vectorize Zoledronate, an anti-cancer interest molecule of the hydroxmethylenebisphosphonate's family. In fact, Zoledronate is a powerful adjuvant in the treatment of bone diseases such as osteoporosis and Paget's disease. But, recent studies have shown that in addition to anti-osteoclastic properties, it presents antitumour properties notably in the case of breast and prostate cancer. However, these properties cannot be exploited due to their very high affinity to divalent cations and their preferentially accumulation in bone. To overcome this problem, one strategy is the vectorization trough maghemite nanocrystal functionalization. The specific surface coating permits to consider γFe₂O₃@Zoledronate as a drug delivery vehicle for therapeutic activity. The anchoring to the nanoparticle's surface allowed to increase their hydrophobicity and also to change the therapeutic target, increasing the Zoledronate intestinal absorption instead of their accumulation in bone. We show that Zoledronate link the nanoparticle surface through phosphonate groups. The biological in vitro tests performed on breast cancer cell line, MDA-MB 231, showed that γFe₂O₃@Zoledronate have antiproliferative activity. In addition, the γFe₂O₃ core could be used as MRI contrast agent for a good therapeutic evaluation.     

The achievement of ligand-functionalized organic/polymeric nanoparticles for treating multidrug resistant cancer

Introduction: The effectiveness of conventional cancer chemotherapy is hampered by the occurrence of multidrug resistance (MDR) in tumor cells. Although many studies have reported the development of novel MDR chemotherapeutic agents, clinical success is lacking owing to the high associated toxicity. Nanoparticle-based delivery of chemotherapeutic drugs has emerged as alternative approach to treat MDR cancers via exploitation of leaky vasculature in the tumor microenvironment. Accordingly, functionalization of nanoparticles with target specific ligands can be employed to achieve significant improvements in the treatment of MDR cancer.

Areas covered: This review focuses on the recent advances in the functionalization of nanocarriers with specific ligands, including antibodies, transferrin, folate, and peptides to overcome MDR cancer. The limitations of effective ligand-functionalized nanoparticles as well as therapeutic successes in ligand targeting are covered in the review.

Expert opinion: Targeting MDR tumors with ligand-functionalized nanoparticles is a promising approach to improve the treatment of cancer. With this approach, higher drug concentrations at targeted sites would be achieved with lower dosage frequencies and reduced side effects in comparison to existing formulations of chemotherapeutic drugs. However, potential toxicities and immunological responses to ligands should be carefully reviewed for viable options in for future MDR cancer treatment.     

3H-咯萘啶在小鼠体内的分布和排泄

咯萘啶(PND)是一种疟原虫红内期裂殖体杀灭剂,与氯喹无交叉抗性,对恶性疟和间日疟均有显著疗效。用家兔的初步实验表明,PND原药在尿中的排泄量仅约为剂量的2%。我们已报告该药在体内分布较广。文献报道PND对鼠疟原虫红内期裂殖体有长效作用。为进一步了解该药的体内过程,对~3H-PND在小鼠体内的分布和排泄进行了研究。     

抗肿瘤药物纳米粒载体的制备材料、包载药物及修饰方法

纳米粒作为抗肿瘤药物的载体,具有提高药物靶向性、稳定性、降低药物毒副作用等诸多优点。近年来,抗肿瘤药物纳米粒载体研究取得了较大进展,从其制备材料、包载药物及修饰方法3方面进行综述。     

Flocculated Amorphous Nanoparticles for Highly Supersaturated Solutions

Purpose  To recover polymer-stabilized amorphous nanoparticles from aqueous dispersions efficiently by salt flocculation and to show that the particles redisperse and dissolve rapidly to produce highly supersaturated solutions. Methods  Nanoparticle dispersions of itraconazole stabilized by nonionic polymers were formed by antisolvent precipitation and immediately flocculated with sodium sulfate, filtered and dried. The size after redispersion in water, crystallinity, and morphology were compared with those for particles produced by spray drying and rapid freezing. Results  Particle drug loading increased to ∼90% after salt flocculation and removal of excess polymer with the filtrate. The formation of the flocs at constant particle volume fraction led to low fractal dimensions (open flocs), which facilitated redispersion in water to the original primary particle size of ∼300 nm. Amorphous particles, which were preserved throughout the flocculation–filtration–drying process, dissolved to supersaturation levels of up to 14 in pH 6.8 media. In contrast, both spray dried and rapidly frozen nanoparticle dispersions crystallized and did not produce submicron particle dispersions upon addition to water, nor high supersaturation values. Conclusions  Salt flocculation produces large yields of high surface area amorphous nanoparticle powders that de-aggregate and dissolve rapidly upon redispersion in pH 6.8 media, for supersaturation levels up to 14.     

治疗炎症性肠病的纳米颗粒研究进展

炎症性肠病是一组慢性非特异性肠道炎症性疾病,目前临床上常用的传统非靶向药物可以缓解患者症状,但不能终止黏膜炎症活动、疾病发展,且严重的不良反应限制了这些药物的应用。纳米颗粒由于其独特的大小和物理特性,能够通过黏液层将负载药物传递到肠道细胞,也可以通过内吞作用被巨噬细胞吞噬,从而调节肠道的免疫环境,此外,纳米颗粒还具有改变药物特性的潜力。总结了治疗炎症性肠病的合成纳米颗粒、天然纳米颗粒的研究情况,以期为炎症性肠病的治疗带来新的机遇。     

Nanoparticulate carrier system: a novel treatment approach for hyperlipidemia

Hyperlipidemia is a prevailing risk factor that leads to development and progression of atherosclerosis and consequently cardiovascular diseases. Several antihyperlipidemic drugs are having various disadvantages such as low water solubility and poor bioavailabilty due to presystemic gastrointestinal clearance. Thus, there is a considerable need for the development of efficient delivery methods and carriers. This review focuses on the importance and role of various nanoparticulate systems as carrier for antihyperlipidemic drugs in the treatment of hyperlipidemia. Some nanoparticle technology-based products are approved by FDA for effective treatment of hyperlipidemia, namely Tricor® by Abbott Laboratories (Chicago, IL, USA) and Triglide® by Skye Pharma (London, UK). Efforts to address each of these issues are going on, and should remain the focus on the future studies and look forward to many more clinical products in the future.     

Anti-inflammatory and Antioxidant Activity of Ageratum conyzoides

Ageratum conyzoides L., a plant widely used in African and South American folk medicine, contains many active principles, including pyrrolizidine alkaloids and polymethoxyflavones. We undertook the present study to evaluate the effect of the methanol extract and of the flavonoid fraction of the aerial part of the plant on carrageenan-induced edema in rat. We subjected the methanol extract to column chromatography to separate the flavonoids and assayed the radical scavenging activity of this fraction by the DPPH method. The two preparations produced significant inhibition on carrageenan-induced rat paw edema, until 2 h after carrageenan treatment. The flavonoids exhibited a strong inhibitory activity on the DPPH radical. The anti-inflammatory effect of A. conyzoides methanol extract depends on the flavonoid fraction, which could produce a protective action against free-radical mediated damage in cells and tissue. Therefore, it is possible to hypothesize that flavonoids influence inflammatory gene protein expression.     

DNA/RNA-based formulations for treatment of breast cancer

Introduction: To develop a successful formulation for the gene therapy of breast cancer, an effective therapeutic nucleic acid and a proper delivery system are essential. Increased understanding of breast cancer, and developments in biotechnology, material science and nanotechnology have provided a major impetus in the development of effective formulations for the gene therapy of breast cancer.

Areas covered: We discuss DNA/RNA-based formulations that can inhibit the growth of breast cancer cells and control the progress of breast cancer. Targets for the gene therapy of breast cancer, DNA/RNA-based therapeutics and delivery systems are summarized. And examples of successful DNA/RNA-based formulations for breast cancer gene therapy are reviewed.

Expert opinion: Several challenges remain in developing effective DNA/RNA-based formulations for treatment of breast cancer. Firstly, most of the currently utilized targets are not effective enough as monotherapy for breast cancer. Secondly, the requirements for co-delivery system make the preparation of formulation more complicated. Thirdly, nanoparticles with the modification of tumor-targeting ligands could be more unstable in circulation and normal tissues. Lastly, immune responses against the viral vectors are unfavorable for the gene therapy of breast cancer because of the damage to the host and the impaired therapeutic ability.     

Utility of certain π-acceptors for the spectrophotometric determination of perindopril

Simple, rapid, accurate and sensitive spectrophotometric methods are described for the determination of perindopril. The methods are based on the reaction of this drug as n-electron donor with 2,3-dichloro-5,6-dicyano-p-benzoquinone(DDQ)-7,7,8,8-tetracyanoquinodimethane (TCNQ), tetracyanoethylene (TCNE), chloranil (CL) and p-chloranilic acid (p-CA) as π-acceptors to give highly coloured complex species. The coloured products are measured spectrophotometrically at 588, 843, 419, 550 and 520 nm for DDQ, TCNQ, TCNE, CL and p-CA, respectively, optimization of different experimental conditions is described. Beer's law is obeyed in the range of 20–200 μg ml⁻¹ and colours were produced in non-aqueous media and were stable for at least 1 h. Application of the suggested methods to perindopril tablets are presented.