雷公藤甲素聚乳酸纳米粒的制备及毒性

目的探索可生物降解聚乳酸［poly(D,L-lactic acid)，PLA］纳米粒口服给药后降低毒性的可能性。方法 采用改良的自乳化溶剂蒸发法制备雷公藤甲素聚乳酸纳米粒；透射电子显微镜(TEM)观察纳米粒的形态；动态激光粒度分析仪测定其平均粒径大小和分布；采用反相高效液相色谱法(RP-HPLC)测定纳米粒的包封率及载药量；X-射线粉末衍射(X-ray)初步研究纳米粒中药物的物理状态；考察雷公藤甲素的体外释放特性；评价口服给予纳米粒对大鼠的降毒性作用。 结果确定适合处方的工艺为：水相-有机相为40∶15(v/v)，表面活性剂浓度为1% (w/v)，药物在有机相中的浓度为0.3% (w/w)，TP-PLA为1∶15 (w/w)。处方条件下制备的纳米粒平均粒径为149.7 nm，多分散指数为0.088，平均包封率及载药量分别为74.27% 和1.36%；雷公藤甲素的体外释放分为两相；纳米粒非常显著降低肝的毒性(P<0.01)，显著降低肾的毒性(P<0.05)。结论聚乳酸纳米粒可能成为雷公藤甲素口服给药的新型载体。     

新型阿霉素抗耐药性隐形脂质体的体外细胞毒和体内毒性研究

目的多药耐药(multidrug resistance,MDR)是目前临床肿瘤治疗的主要障碍。本文研制了新型阿霉素抗耐药性隐形脂质体(DARSLs)，并对其体外细胞毒和体内毒性进行评价。方法采用硫酸铵梯度法将阿霉素(DOX)和维拉帕米(VER)药物同时包载到隐形脂质体内，制备成DARSLs；采用耐药性鼠前列腺肿瘤细胞株MLLB2和人子宫肉瘤细胞株MES-SA/DX5进行体外细胞毒性评价；采用SD大鼠对阿霉素抗耐药性隐形脂质体进行体内毒性评价。结果在药脂比(DOX/VER/Lipid，w/w/w)为1∶0.11∶10时，阿霉素包封率大于90%，维拉帕米包封率约为70%。平均粒径为(118.1±22.3) nm。体外细胞毒性实验证实该脂质体能够在体外有效地逆转肿瘤细胞耐药性，并导致耐药肿瘤细胞生长抑制。体内系统毒性及心脏毒性实验结果显示，该脂质体能够明显改善游离阿霉素单独使用或与维拉帕米联合使用时产生的全身毒性，尤其是心脏毒性。结论DARSLs具有相对较低的毒性，且能有效抑制耐药肿瘤的生长。     

醋酸地塞米松固体脂质纳米粒的制备及表征

以山嵛酸甘油酯(Compdtol 888 ATO)为载体,采用热融-超声法制备醋酸地塞米松固体脂质纳米粒.制品平均粒径(106.8±6.8)nm,DSC结果表明药物包载在纳米粒中.稳定性初步研究显示纳米粒的外观和包封率在3个月内较稳定.体外释放试验表明,纳米粒在pH 7.4磷酸盐缓冲液释药前期有一定量的突释,后期则具有明显的缓释特征,释放时间可达6d.     

重组人血管内皮抑制素壳聚糖纳米粒的制备

目的 以壳聚糖(CS)为载体,研究其对重组人血管内皮抑制素(Endostar)的包封及控释能力.方法采用大分子复合法制备纳米粒,考察形成条件及冻干工艺,并对纳米粒的形态、包封率、体外释放及Endostar的完整性进行考察.结果CS与羧甲基纤维素钠( CMC - Na)的质量比介于6∶1～1∶2时可以形成纳米粒,粒径为1...     

甲氨蝶呤叶酸受体-磁双重靶向纳米粒的制备及评价

目的：以甲氨蝶呤为药物模型，制备用于肿瘤靶向治疗的叶酸受体-磁双重靶向纳米药物。方法：未采用 预成型的磁性纳米粒，一步合成磁性纳米粒核二氧化硅壳超顺磁性的纳米粒，并借助透射、扫描电镜观察微球形态，用 硅烷偶联剂进行表面修饰，在表面化学偶联上叶酸，修饰甲氨蝶呤后利用紫外可见分光光度计测量载药量及包封率。结 果：磁性纳米粒在电镜下呈现核壳样球型微粒，平均粒径为20 nm，纳米粒载药量为26.71%，包封率为64.76%。结论： 叶酸受体-磁双重载药纳米粒为肿瘤的靶向治疗提供了一种可能的新剂型，有较好的临床应用前景。     

用注射法和复乳法制备低分子肝素脂质纳米粒

目的:用注射法和复乳法制备了两种亲水性药物低分子肝素脂质纳米粒.方法:运用TEM和LD等方法,对纳米载体的包封率、平均粒径及粒径分布和形态学差异进行了研究.结果:注射法(115 nm)制备脂质纳米粒的粒径要小于复乳法(273 nm),但后一种方法可以实现较好的包封,包封率高达37%.并能观察到较为完全的W/O/W的复合包封结构.结论:用复乳法制备肝素脂质纳米粒可以获得较好的包封效果.     

阿柔比星A PELGE纳米粒的制备及质量评价

目的研究阿柔比星A PELGE纳米粒的制备及质量评价。方法通过共沉淀法制备阿柔比星A PELGE纳米粒；考察阿柔比星A PELGE纳米粒的形态、粒径、包封率、载药量、Zeta电位和体外释药。结果阿柔比星A PELGE纳米粒在透射电镜下均呈圆球形，分布均匀，平均粒径为105～137nm，平均包封率为76％～86％，平均载药量为7．6％～8．6％，平均Zeta电位为-25～-33mV。纳米粒的体外释药曲线可分为突释和缓释两部分，体外释药曲线以Weibull方程拟合为好。PELGE共聚物中的PEG分子量和百分量改变对阿柔比星APELGE纳米粒的粒径、包封率、载药量、Zeta电位和释药速率的影响不明显。结论PELGE纳米粒作为传输药物的载体是可行的，并为PELGE纳米粒的进一步研究提供了实验依据。     

苦参素磷脂纳米粒的研制

目的:制备稳定性好、亲和力强的苦参素大豆磷脂纳米粒(KU-PL-NP).方法:利用大豆磷脂为载体,采用薄膜-超声分散法制备了苦参素大豆磷脂纳米粒,以制剂的含量测定、载药量、包封率及粒径为主要评价指标.结果:3批制剂平均包封率为64.05%,平均载药量为21.4%,平均粒径为309.5 nm.结论:苦参素大豆磷脂纳米粒基本达到设计要求,该制剂有望成为一种新的药物靶向载体系统.     

壳聚糖孵育低分子肝素钙脂质体的制备及其包封率测定

目的:尝试通过壳聚糖孵育以提高低分子肝素钙脂质体包封率,并分析其可能机制。方法:采用薄膜分散法制备低分子肝素钙脂质体,1∶5壳聚糖溶液孵育低分子肝素钙脂质体。利用天青A法测定包封率。结果:低分子肝素钙脂质体经壳聚糖孵育前、后的包封率分别为(46.40±3.14)%、(97.94±1.33)%。透射电镜照片示镜下有纳米粒生成。结论:壳聚糖孵育的低分子肝素钙脂质体包封率大幅提高,可能是由于壳聚糖与游离的低分子肝素钙形成纳米粒所致。     

阿霉素壳寡糖纳米粒的制备及体外抗肿瘤研究

目的制备负载阿霉素的壳寡糖纳米粒,并研究其理化性质和体外抗肿瘤细胞毒性。方法采用离子凝胶法制备负载阿霉素的壳寡糖纳米粒;透射电镜观察纳米粒形态,激光粒度仪测定粒径和表面电位,紫外分光光度法测量包封率、载药量,考察载药纳米粒的体外释药特性;采用MTT法对载药壳寡糖纳米粒在体外乳腺癌细胞株MCF-7的细胞毒作用进行评价。结果制得的阿霉素壳寡糖纳米粒呈球形或类球形,形态较为完整,平均粒径为(136.77±1.21)nm,表面电位为(20.53±0.31)m V,包封率为(56.99±1.40)%,载药量为(15.49±0.38)%,168 h的累积释放率为72.15%;阿霉素和载药纳米粒对MCF-7细胞增殖的抑制作用存在明显的浓度和时间依赖性,且载药纳米粒对MCF-7细胞增殖的抑制作用随时间增加而逐渐强于游离阿霉素。结论此方法制备的阿霉素壳寡糖纳米粒粒径较小,药物释放具有明显的缓释作用,并具有较好的抗肿瘤作用。     

结肠癌细胞及线粒体双级靶向脂质体的处方工艺筛选研究

目的：研究结肠癌细胞及线粒体双级靶向脂质体（HA/TPP-TPGS LP/DOX）的最佳处方工艺。方法：用薄膜分散法结合微孔滤膜法制备;以细胞抑制率为指标,用MTT法筛选最佳聚脂比;以包封率为指标,用正交试验筛选最佳胆脂比、药脂比和超声时间;以粒径为指标,筛选最佳透聚比;以复溶后粒径和包封率为指标,筛选冻干保护剂的品种;用荧光显微镜和流式细胞术考察脂质体的靶向性;用透析法考察体外释药行为。结果：最佳处方是聚脂比1∶7、胆脂比1∶10、药脂比1∶15、超声时间15 min、透聚比2∶1,冻干保护剂为蔗糖。制备的脂质体呈类球形,粒径（142.20±0.54）nm,Zeta电位-（24.06±0.25）mV,包封率（98.20±0.18）%,稳定性高,有双级靶向性和体外药物缓释性。结论：本研究制备的脂质体有包封率高、粒径小、双级靶向性和缓释性等优点,为进一步研究奠定了基础。     

麦角甾苷固体脂质纳米粒处方筛选和理化性质研究

目的 研究不同种类药用辅料成分对麦角甾苷固体脂质纳米粒（SLN）理化性质的影响,为研究SLN的处方筛选提供依据。方法 采用乳化-固化法制备麦角甾苷-SLN,单一变量法考察山嵛酸甘油酯（Compritol ATO 888）、单硬脂酸甘油酯、大豆卵磷脂、Myrj52等辅料对麦角甾苷-SLN粒径、包封率、表征分散度（PDI）等理化性质的影响,采用透射电镜法观察麦角甾苷-SLN的形态,X-射线衍射（XRD）分析其药物晶体结构。结果 随Compritol ATO 888用量增加,麦角甾苷-SLN粒径不断减小,包封率逐渐减小,PDI逐渐增加;随单硬脂酸甘油酯的用量增加,粒径明显增大,包封率略有降低,PDI减小;随卵磷脂用量增加,粒径明显增大,包封率降低,PDI减小;随Myrj52用量明显增加,粒径减小,包封率增加,PDI增大;麦角甾苷-SLN外观圆整,呈球形;麦角甾苷以分子分散状态被包裹在SLN中。结论 不同辅料对麦角甾苷-SLN的理化性质均产生一定影响趋势,为制备SLN的处方筛选研究提供启示与思路。     

Cascade two-stage tumor re-oxygenation and immune re-sensitization mediated by self-assembled albumin-sorafenib nanoparticles for enhanced photodynamic immunotherapy

As a promising modality for cancer therapy, photodynamic therapy (PDT) still acquired limited success in clinical nowadays due to the extremely serious hypoxia and immunosuppression tumor microenvironment. To ameliorate such a situation, we rationally designed and prepared cascade two-stage re-oxygenation and immune re-sensitization BSA-MHI148@SRF nanoparticles via hydrophilic and hydrophobic self-assembly strategy by using near-infrared photodynamic dye MHI148 chemically modified bovine serum albumin (BSA-MHI148) and multi-kinase inhibitor Sorafenib (SRF) as a novel tumor oxygen and immune microenvironment regulation drug. Benefiting from the accumulation of SRF in tumors, BSA-MHI148@SRF nanoparticles dramatically enhanced the PDT efficacy by promoting cascade two-stage tumor re-oxygenation mechanisms: (i) SRF decreased tumor oxygen consumption via inhibiting mitochondria respiratory. (ii) SRF increased the oxygen supply via inducing tumor vessel normalization. Meanwhile, the immunosuppression micro-environment was also obviously reversed by two-stage immune re-sensitization as follows: (i) Enhanced immunogenic cell death (ICD) production amplified by BSA-MHI148@SRF induced reactive oxygen species (ROS) generation enhanced T cell infiltration and improve its tumor cell killing ability. (ii) BSA-MHI148@SRF amplified tumor vessel normalization by VEGF inhibition also obviously reversed the tumor immune-suppression microenvironment. Finally, the growth of solid tumors was significantly depressed by such well-designed BSA-MHI148@SRF nanoparticles, which could be potential for clinical cancer therapy.     

Sharp pH-sensitive amphiphilic polypeptide macrophotosensitizer for near infrared imaging-guided photodynamic therapy

Tumor environmental sensitive polypeptide integrated photosensitizer is a platform for imaging-guided photodynamic therapy (PDT). However, the photosensitizer leakage during blood circulation, poor accumulation in tumor tissue and inferior quantum yield of singlet oxygen are still challenges. Herein, NHS-active boron-dipyrromethene derivative with bromine substituted NHS-BODIPY-Br₂ was first synthesized, which possessed high singlet oxygen generation efficiency and near infrared (NIR) fluorescence, and then it was conjugated to a sharp pH (6.36) sensitive polypeptide to achieve a macrophotosensitizer for NIR imaging-guided PDT. In vitro study showed that the macrophotosensitizer nanoparticles exhibited good cellular uptake and ability to kill cancer cells. Once accumulating in the tumor tissues, the nanoparticles can be demicellized by tumor acidity to promote cellular uptake, which could enlarge fluorescence signal intensity and enhance in vivo PDT therapeutic effect upon NIR laser irradiation. It provides a strategy to design photosensitizer conjugated tumor acidity sensitive polypeptide for NIR imaging-guided photodynamic therapy.     

Nanoparticle-mediated combination chemotherapy and photodynamic therapy overcomes tumor drug resistance in vitro

Drug resistance limits the success of many anticancer drugs. Reduced accumulation of the drug at its intracellular site of action because of overexpression of efflux transporters such as P-glycoprotein (P-gp) is a major mechanism of drug resistance. In this study, we investigated whether photodynamic therapy (PDT) using methylene blue, also a P-gp inhibitor, can be used to enhance doxorubicin-induced cytotoxicity in drug-resistant tumor cells. Aerosol OT (AOT)-alginate nanoparticles were used as a carrier for the simultaneous cellular delivery of doxorubicin and methylene blue. Methylene blue was photoactivated using light of 665 nm wavelength. Induction of apoptosis and necrosis following treatment with combination chemotherapy and PDT was investigated in drug-resistant NCI/ADR-RES cells using flow cytometry and fluorescence microscopy. Effect of encapsulation in nanoparticles on the intracellular accumulation of doxorubicin and methylene blue was investigated qualitatively using fluorescence microscopy and was quantitated using HPLC. Encapsulation in AOT-alginate nanoparticles significantly enhanced the cytotoxicity of combination therapy in resistant tumor cells. Nanoparticle-mediated combination therapy resulted in a significant induction of both apoptosis and necrosis. Improvement in cytotoxicity could be correlated with enhanced intracellular and nuclear delivery of the two drugs. Further, nanoparticle-mediated combination therapy resulted in significantly elevated reactive oxygen species (ROS) production compared to single drug treatment. In conclusion, nanoparticle-mediated combination chemotherapy and PDT using doxorubicin and methylene blue was able to overcome resistance mechanisms and resulted in improved cytotoxicity in drug-resistant tumor cells.     

Surfactant-polymer nanoparticles enhance the effectiveness of anticancer photodynamic therapy

Photodynamic therapy (PDT) is a promising treatment modality for cancer. PDT is based on the concept that photosensitizers, when exposed to light of specific wavelength, generate cytotoxic reactive oxygen species (ROS) capable of killing tumor cells. The effectiveness of PDT has been limited in part by the lack of photosensitizers that accumulate sufficiently in tumor cells and poor yield of ROS from existing photosensitizers. In this report, we investigated whether aerosol OT-alginate nanoparticles can be used as a carrier to enhance the therapeutic efficacy of a model photosensitizer, methylene blue. Methylene blue loaded nanoparticles were evaluated for PDT effectiveness in two cancer cell lines, MCF-7 and 4T1. Encapsulation of methylene blue in nanoparticles significantly enhanced intracellular ROS production, and the overall cytotoxicity following PDT. It also resulted in higher incidence of necrosis. Greater effectiveness of nanoparticles could be correlated with higher yield of ROS with nanoparticle-encapsulated methylene blue. Further, treatment of tumor cells with nanoparticle-encapsulated methylene blue resulted in significant nuclear localization of methylene blue while free drug treatment resulted in its accumulation mainly in the endolysosomal vesicles. In conclusion, encapsulation of methylene blue in aerosol OT-alginate nanoparticles enhanced its anticancer photodynamic efficacy in vitro. Increased ROS production and favorable alteration in the subcellular distribution contribute to the enhanced PDT efficacy of nanoparticle-encapsulated photosensitizer.     

薄膜超声法制备槲皮素脂质体研究

目的 以粒径和包封率为指标,优选槲皮素脂质体的制备工艺。 方法 以氢化大豆磷脂(HSPC)、胆固醇(CH)为膜材,采用薄膜超声法制备脂质体。通过正交设计优化处方工艺,利用马尔文动态光散射粒径测定仪测定脂质体的粒径,鱼精蛋白沉淀法分离游离药物,HPLC法测定脂质体中槲皮素(QU)的包封率。 结果 最佳处方工艺为:HSPC:CH=3:1、HSPC:QU=20:1、探头超声(600 W)9 min。 结论 薄膜超声法适于实验室条件下制备槲皮素脂质体。     

维A酸脂质体的制备工艺研究

目的:提供一种制备稳定维A酸脂质体的新方法。方法:采用乙醇注入法制备维A酸脂质体,通过单因素试验和四因素三水平正交试验,以包封率为主要指标,优选药脂比、磷脂与胆固醇之比、水合介质(0.01 mol/L磷酸盐缓冲液)的pH和用量;并制备样品进行稳定性考察。结果:以药脂比为1∶10(维A酸用量1.0 mg),磷脂与胆固醇之比为4∶1,水合介质的pH为6.5、用量为30ml为最佳工艺;所制维A酸脂质体的包封率约为80%,平均粒径约为150 nm。在室温条件下密封放置10 d及4℃下保存6个月,其平均粒径及包封率差异无统计学意义(P>0.05)。结论:该制剂制备工艺简单可行;制剂处方合理,包封率较高,且在短期内稳定性良好。     

Ir(III) Complex Dimer Nanoparticles for Photodynamic Therapy

Iridium compounds are versatile in catalysis and photodynamic therapy. We present two Ir(III) complex dimers that can self-assemble into nanoparticles in the absence of adjuvant or surfactants. The formed nanoparticles possess a spherical morphology and robust colloidal stability and could be internalized by cancer cells. The Ir(III) complex nanoparticles have relatively strong photodynamic activity upon irradiation, which includes type I and type II reactive oxygen species. The generated reactive oxygen species could effectively induce cell death upon irradiation. This work highlights the potential of metal complexes and their nanoparticles in cancer treatment.     

Tailored core‒shell dual metal–organic frameworks as a versatile nanomotor for effective synergistic antitumor therapy

Malignant tumor has become an urgent threat to global public healthcare. Because of the heterogeneity of tumor, single therapy presents great limitations while synergistic therapy is arousing much attention, which shows desperate need of intelligent carrier for co-delivery. A core‒shell dual metal–organic frameworks (MOFs) system was delicately designed in this study, which not only possessed the unique properties of both materials, but also provided two individual specific functional zones for co-drug delivery. Photosensitizer indocyanine green (ICG) and chemotherapeutic agent doxorubicin (DOX) were stepwisely encapsulated into the nanopores of MIL-88 core and ZIF-8 shell to construct a synergistic photothermal/photodynamic/chemotherapy nanoplatform. Except for efficient drug delivery, the MIL-88 could be functioned as a nanomotor to convert the excessive hydrogen peroxide at tumor microenvironment into adequate oxygen for photodynamic therapy. The DOX release from MIL-88-ICG@ZIF-8-DOX nanoparticles was triggered at tumor acidic microenvironment and further accelerated by near-infrared (NIR) light irradiation. The in vivo antitumor study showed superior synergistic antitumor effect by concentrating the nanoparticles into dissolving microneedles as compared to intravenous and intratumoral injection of nanoparticles, with a significantly higher inhibition rate. It is anticipated that the multi-model synergistic system based on dual-MOFs was promising for further biomedical application.