基于普朗尼克双配体修饰的载药纳米靶向聚合物胶束制备工艺的筛选及靶向性评价

目的 优选载多柔比星(阿霉素,DOX)纳米靶向聚合物胶束的制备工艺,并对其体外释放行为和体外靶向性进行考察.方法 以琥珀酰亚胺法合成靶向聚合物材料,以包封率、载药量和总评归一化值为评价指标,利用星点设计-响应面法优化薄膜水化法设计考察投药量、有机溶剂体积、水化体积对纳米胶束制备的影响,并优选处方.以与羟基磷灰石(HA)和离体骨片共同孵育考察骨靶向性;以乳腺癌细胞(MDA-MB-231)为模型做体外细胞摄取考察肿瘤细胞靶向性.结果 成功合成靶向聚合物材料P123-ALN和P123-DP-8,优选纳米胶束最佳制备工艺为:DOX投药量5.48 mg,有机溶剂体积7.28 ml,水化体积8.58 ml.根据筛选出的最佳制备工艺,确定混合载体材料的比例为4∶1时,制备的双配体修饰的纳米靶向聚合物胶束P123-ALN/P123-DP-8@DOX符合纳米制剂的制备要求,包封率为76.97％,载药量3.70％,粒径122.97 nm,ζ电位-12.60 mV,缓释和体外靶向性良好.结论 用最优处方制备的纳米靶向聚合物胶束可为后续骨靶向给药奠定基础.     

多西他赛热敏脂质体的制备及含量和包封率的测定

目的制备多西他赛（DTX）热敏脂质体（docetaxel-loaded thermosensitive liposomes,DTL）,建立其含量和包封率测定方法。方法采用薄膜分散法制备DTL并用高效液相色谱法（HPLC）测定脂质体中多西他赛的含量。采用高速离心法分离脂质体与游离药物并测定包封率。结果制备的DTL粒径均一,平均粒径为104 nm,相变温度为42.20℃。辅料和溶剂不干扰主药含量测定,线性范围为1.0～100.0μg/ml（r=1）;多西他赛平均回收率为99.50%（RSD=0.65%）。药物在脂质体中的包封率大于98%。结论薄膜分散法可用于制备DTL。药物含量和包封率测定方法简便快速,准确可靠,专属性强。     

多西他赛热敏脂质体的制备及含量和包封率的测定

目的制备多西他赛(DTX)热敏脂质体(docetaxel-loaded thermosensitive liposomes,DTL),建立其含量和包封率测定方法。方法采用薄膜分散法制备DTL并用高效液相色谱法(HPLC)测定脂质体中多西他赛的含量。采用高速离心法分离脂质体与游离药物并测定包封率。结果制备的DTL粒径均一,平均粒径为104 nm,相变温度为42.20℃。辅料和溶剂不干扰主药含量测定,线性范围为1.0～100.0μg/ml(r=1);多西他赛平均回收率为99.50%(RSD=0.65%)。药物在脂质体中的包封率大于98%。结论薄膜分散法可用于制备DTL。药物含量和包封率测定方法简便快速,准确可靠,专属性强。     

度他雄胺纳米悬浮液的制备

目的 运用高压均质技术制备度他雄胺纳米悬浮液.方法 以粒径、多分散性和Zeta电位为考察指标,研究均质压力、循环次数、表面活性剂种类和浓度等因素对纳米悬浮液性质的影响,并考察度他雄胺的溶解度及药物溶出速率.结果 均质压力是影响纳米粒径的主要因素.纳米尺度的度他雄胺的溶解度和体外溶出速率比原药明显提高.结论 高压均质技术可以成功用于制备度他雄胺纳米悬浮液.     

携药靶向前列腺癌的聚吡咯超声/荧光微泡的制备及其特性研究

目的 制备载多西他赛、吲哚菁绿的聚吡咯纳米壳微泡超声造影剂,评价其理化特性并考察其体外寻靶能力。方法 应用机械震荡法制备携药聚吡咯超声/荧光造影剂。检测并观察造影剂微泡的形态、造影剂粒径和表面电位。将造影剂放置在4℃保存,在1天、3天、5天不同的时间观察造影剂的稳定性。应用显微镜观察前列腺癌细胞摄取吲哚菁绿的情况。结果 新制备的造影剂外观圆整,颗粒直径范围为875～1516 nm,表面电位为(-22.4±1.75) m V,分布较均匀。在制备3天后,密封保存在4℃的造影剂浓度缓慢的下降,5天后其浓度明显下降。显微镜观察到前列腺癌细胞摄取吲哚菁绿成像呈绿色荧光。结论 制备的携药聚吡咯超声/荧光微泡相关理化性能较好,稳定性较高,能够被前列腺癌细胞摄取,为前列腺癌的精确诊断和光热消融提供了新的思路。     

促黄体激素释放激素a(LHRHa)靶向鸦胆子油脂质体的制备及质量评价

目的 制备促黄体激素释放激素a(LHRHa)靶向鸦胆子油脂质体并评价其质量.方法 采用薄膜分散法结合生物素-链霉亲和素桥接法制备LHRHa靶向鸦胆子油脂质体,正交设计优选处方,透射电镜下观察脂质体形态,采用Zetasizer Nano ZS分析仪测定脂质体粒径、Zeta电位,葡聚糖凝胶柱层析结合分光光度法测定包封率,通过离心加速实验及渗漏率考察脂质体稳定性,体外细胞实验鉴定脂质体靶向性.结果 所得优化处方为磷脂与胆固醇比4∶1,药物与脂质比3∶10,DSPE-PEG-Biotin与卵磷脂质量比为3％,超声乳化时间为8min.按此处方制备的脂质体形态圆整,粒径为155.1±14.5nm,Zeta电位为-24.1±0.54mV,平均包封率达92.2％,对卵巢癌A2780/DDP细胞的亲和力约为普通脂质体的2.7倍.结论 LHRHa靶向鸦胆子油脂质体制备工艺可行,同时具有包封率高、稳定性及靶向性良好等优点.     

注射用紫杉醇亚微乳的制备及其性质考察

目的制备静脉注射用紫杉醇亚微乳并考察其理化性质。方法采用高压匀质法制备紫杉醇亚微乳,考察处方和工艺主要因素对乳剂相关性质的影响,评价其质量,并对血管刺激性进行考察。结果优化条件下制备的紫杉醇亚微乳粒度分布均匀,平均粒径为（168±22）nm,多分散系数为（0.89±0.05）,Zeta电位为-（33.9±0.5）mV,稳定性较好,血管刺激性小。结论紫杉醇亚微乳处方和工艺合理可靠,安全性好,便于生产和临床使用。     

雷替曲塞联合多西他赛治疗晚期胃癌的临床研究

目的观察雷替曲塞联合多西他赛方案治疗晚期胃癌的近期疗效和毒副作用。方法选择40例经病理组织学诊断的晚期胃癌,采用雷替曲塞联合多西他赛方案化疗,其中雷替曲塞（商品名赛维健,南京正大天晴制药有限公司生产）2．5mg／m2,15min内静脉推注,第1天,联合多西他赛75mg／m2,60min静脉滴注,第1,8,15天,每3周为1周期;2个周期后评价疗效。结果40例总有效21例（52．5％）,其中CR2例,占5％;PR19例,占47．5％;S1915例,占37．5％;PD6例,占15％。主要的毒副作用为骨髓抑制、恶心呕吐、口腔黏膜炎、腹泻、乏力、关节疼痛及水钠潴留等,全组未见肝。肾功能和心脏损害,无治疗相关性死亡患者．也无因毒性反应而延缓化疗者。结论雷替曲塞联合多西他赛方案治疗晚期胃癌疗效较好,毒副反应能耐受,值得临床推广使用。     

阿霉素磷脂毫微粒复合体的制备及药剂学特征研究

目的　制备阿霉素磷脂毫微粒复合体。方法　采用均匀设计等方法筛选处方和工艺。先制备阿霉素聚氰基丙烯酸正丁酯毫微粒 ,再采用磷脂双分子层包裹 ,得到阿霉素磷脂毫微粒复合体。结果　制得的磷脂毫微粒复合体的平均包封率为(86 .6 5± 0 .97% ) ,平均粒径为 2 5 6 .2nm。稳定性较普通脂质体好 ,包封率比普通毫微粒高。结论　阿霉素磷脂毫微粒复合体达到设计要求 ,有可能成为一种新的药物靶向载体给药系统     

多西他赛联合吡柔比星治疗晚期乳腺癌32例临床分析

目的：观察多西他赛联合吡柔比星治疗晚期乳腺癌的临床疗效及毒性反应。方法：32例晚期乳腺癌患者,接受多西他赛70 mg/m2,静脉滴注,第1 d,THP 60 mg/m2,快速静滴,第1 d,21 d一周期,总共观察3个周期。结果：多西他赛联合THP的总有效率（CR＋PR）为56.25%,最严重的不良反应是脱发和白细胞减少。结论：西他赛联合THP治疗晚期乳腺癌临床疗效较好,其引起的不良反应可以耐受,因此联合使用多西他赛和THP是晚期乳腺癌治疗的有效方案,值得临床推广。     

Zinc Phthalocyanine loaded polymeric micelles for photodynamic therapy

Photodynamic therapy (PDT) is a promising alternative to currently employed cancer treatments [1]. However, the high hydrophobicity of the photosensitizers (e.g. zinc phthalocyanine (ZnPC)) causes its aggregation in biological fluids. Moreover, unspecific accumulation in healthy tissues such as the skin, and low clearance rate of ZnPC, leads to prolong skin sensitization, forcing patients with a short life expectancy to remain inside and shielded from light. Consequently, clinical implementation is still limited. In this study, poly(caprolactone)n-poly(ethylene glycol) (pCL-PEG) micelles encapsulating ZnPC are being investigated to increase the solubility of ZnPC and its accumulation at the tumor site.pCLn-PEG (n=27) was synthesized by ring-opening polymerization following the protocol from Y. Liu et al. [2]. pLC-PEG micelles encapsulating ZnPC (ZnPC-M) at different drug loadings (0.2, 0.4 and 0.6 % w/w) were prepared by film hydration. Z-average hydrodynamic diameter and size distribution were measured by dynamic light scattering. In vitro stability and ZnPC transfer to human proteins and lipoproteins was compared between ZnPC-M with different drug loadings using asymmetric flow field flow fractionation after incubation of ZnPC-M with human plasma (1:2 v/v) for different timepoints. Phototoxicity of ZnPC-M was tested in vitro by exposing TFK1 (both in 2D and 3D) to a range of ZnPC concentrations. After incubation and washing, the cells were illuminated with a specific light intensity using a LED device and subsequently, further incubated overnight. The cell viability was measured with MTS assay and CellTiter-Glo® 3D Cell Viability Assay. Furthermore, association ZnPC with TFK1 cells was evaluated by fluorescent spectroscopy.The average hydrodynamic size of ZnPC-M increased with amount of ZnPC encapsulated, ranging from 56 ± 1 nm to 70 ± 1 nm. After incubation with human plasma, ZnPC was immediately and partly partitioned to lipoproteins but not to albumin. At 6 hours timepoint, less than 30% of the ZnPC content had been released from the ZnPC-M. The phototoxicity of ZnPC-M in TFK1 cells was inversely correlated with ZnPC loading for both 2D and 3D cells: 0.2% w/w ZnPC-M showed the lowest LC50. Quantification of the ZnPC associated with TFK1 cells before light exposure showed similar amounts of drug independently of the loading (0.2, 0.4 and 0.6 % w/w). These data suggests that ZnPC-M at 0.2% w/w have faster drug release, making the ZnPC available at the time of the illumination for the generation of ROS, and thus inducing higher phototoxicity.pCL-PEG micelles encapsulating ZnPC were developed to improve ZnPC solubility and stability. Increasing drug loading was shown to decrease phototoxicity of ZnPC-M in TFK1 cells, while not affecting plasma stability. Faster drug release seems to be the reasoning for the 0.2% w/w ZnPC-M outperform the higher loaded ZnPC-M. These results prompt further investigation in vivo to confirm the superiority of 0.2% w/w ZnPC-M in both pharmacokinetics and PDT efficacy.     

叶酸受体介导的肿瘤靶向超顺磁性纳米胶束的制备及体外实验

目的探讨叶酸受体介导的两亲聚合物纳米胶束对人肝癌Bel 7402细胞的靶向性，及利用MR仪对其进行监测的可行性。方法制备由叶酸修饰的、载有超顺磁性氧化铁（SPIO）及抗癌药物表阿霉素（DOX）的纳米胶束，将叶酸靶向及非叶酸靶向纳米胶束分别与人肝癌Bel 7402细胞共孵育1h，进行普鲁士蓝染色和流式细胞术观察Bel 7402细胞对叶酸靶向及非叶酸靶向纳米胶束的吸收情况，并体外MRI观察T2WI信号强度变化。结果叶酸靶向纳米胶束与Bel 7402细胞共孵育后普鲁士蓝染色显示细胞内大量铁存在；非叶酸靶向纳米胶束普鲁士蓝染色显示细胞内铁浓度极低。流式细胞术显示叶酸靶向组及非叶酸靶向组的平均荧光强度分别为117．88和46．33，叶酸靶向组约为非叶酸靶向组的2．5倍。体外MRI显示叶酸靶向纳米胶束与Bel 7402细胞共孵育后在T2WI上信号明显降低（SPIO浓度为5、10、20、40和80μg／ml时，信号变化率中位数分别为-5．02％、-23．58％、-45．89％、-70．34％和-92．41％），而非叶酸靶向组在T2WI上信号无明显降低（SPIO浓度为5、10、20、40和80μg／ml时信号变化率中位数分别为-3．77％、-2．16％、-2．18％、-2．74％和-19．77％）。体外竞争抑制实验普鲁士蓝染色显示细胞内铁浓度极低。结论以叶酸修饰的生物可降解聚合物纳米胶束对人肝癌细胞Bel 7402有较好的靶向性，使用临床型MR仪可对其进行监测。     

Chemodosimetry of in vivo tumor liposomal drug concentration using MRI.

Effective cancer chemotherapy depends on the delivery of therapeutic drugs to cancer cells at cytotoxic concentrations. However, physiologic barriers, such as variable vessel permeability, high interstitial fluid pressure, and heterogeneous perfusion, make it difficult to achieve that goal. Efforts to improve drug delivery have been limited by the lack of noninvasive tools to evaluate intratumoral drug concentration and distribution. Here we demonstrate that tumor drug concentration can be measured in vivo using T(1)-weighted MRI, following systemic administration of liposomes containing both drug (doxorubicin (DOX)) and contrast agent (manganese (Mn)). Mn and DOX concentrations were calculated using T(1) relaxation times and Mn:DOX loading ratios, as previously described. Two independent validations by high-performance liquid chromatography (HPLC) and histologic fluorescence in a rat fibrosarcoma (FSA) model indicate a concordant linear relationship between DOX concentrations determined using T(1) and those measured invasively. This method of imaging exhibits potential for real-time evaluation of chemotherapeutic protocols and prediction of tumor response on an individual patient basis.     

高效液相色谱法测定特考韦瑞的平衡溶解度和油水分配系数

目的 用高效液相色谱法(HPLC)测定特考韦瑞(ST-246)在37℃水和不同pH缓冲液的平衡溶解度,以及在正辛醇-水/缓冲液系统的油水分配系数.方法 采用摇瓶法测定平衡溶解度,通过药物分配平衡后在油相及水相的浓度比,计算油水分配系数.结果 ST-246在37℃水的平衡溶解度为3.35 mg/L,ST-246的表观油水分配系数为230.49(lgP=2.36),在正辛醇/pH4.0缓冲液的油水分配系数最大,为779(lgP=2.89).结论 ST-246的水溶性较差,脂溶性不佳,其水中溶解度呈pH依赖性,增大pH可增加其溶解度.     

Proton echo-planar spectroscopic imaging of J-coupled resonances in human brain at 3 and 4 Tesla.

In this multicenter study, 2D spatial mapping of J-coupled resonances at 3T and 4T was performed using short-TE (15 ms) proton echo-planar spectroscopic imaging (PEPSI). Water-suppressed (WS) data were acquired in 8.5 min with 1-cm(3) spatial resolution from a supraventricular axial slice. Optimized outer volume suppression (OVS) enabled mapping in close proximity to peripheral scalp regions. Constrained spectral fitting in reference to a non-WS (NWS) scan was performed with LCModel using correction for relaxation attenuation and partial-volume effects. The concentrations of total choline (tCho), creatine + phosphocreatine (Cr+PCr), glutamate (Glu), glutamate + glutamine (Glu+Gln), myo-inositol (Ins), NAA, NAA+NAAG, and two macromolecular resonances at 0.9 and 2.0 ppm were mapped with mean Cramer-Rao lower bounds (CRLBs) between 6% and 18% and approximately 150-cm(3) sensitive volumes. Aspartate, GABA, glutamine (Gln), glutathione (GSH), phosphoethanolamine (PE), and macromolecules (MMs) at 1.2 ppm were also mapped, although with larger mean CRLBs between 30% and 44%. The CRLBs at 4T were 19% lower on average as compared to 3T, consistent with a higher signal-to-noise ratio (SNR) and increased spectral resolution. Metabolite concentrations were in the ranges reported in previous studies. Glu concentration was significantly higher in gray matter (GM) compared to white matter (WM), as anticipated. The short acquisition time makes this methodology suitable for clinical studies.     

积雪草苷平衡溶解度和油水分配系数的测定

目的测定积雪草苷在不同pH介质中的平衡溶解度和正辛醇-水/缓冲液体系中的表观油水分配系数（Papp）。方法采用RP-HPLC法测定积雪草苷浓度,研究其在水和不同pH介质中的平衡溶解度;采用摇瓶法测定积雪草苷在正辛醇-水/缓冲液体系中的Papp。结果 25℃下积雪草苷在水中的平衡溶解度为305.02μg.ml-1,在pH 5.0缓冲液中平衡溶解度更低;积雪草苷在正辛醇-水中Papp为0.908（logPapp=-0.042）,在pH 5.0缓冲液中Papp最高。结论积雪草苷为亲水性药物,但其在水中的溶解度较小。在pH 5.0～8.0范围内,积雪草苷的平衡溶解度随pH的增加而增加,相反油水分配系数随pH的增加而减少。     

99Tcm-MIBI脂质体的制备及特征分析

目的 研究99Tcm-甲氧基异丁基异腈（MIBI）脂质体的制备方法,并对其特征进行分析。方法 采用乙醇注入-超声法制备99Tcm-MIBI脂质体,测定脂质体粒径大小和包封率,对超声时间、大豆卵磷脂与胆固醇的质量比等条件进行优化,观察99Tcm-MIBI脂质体的稳定性。结果 当制备的99Tcm-MIBI质量浓度为0.01 mg/mL时,平均粒径大小为（171.4±25.2）nm,包封率的平均值为（8.5±1.3）%。大豆卵磷脂与胆固醇的质量比为4:1和超声时间为5 min为最佳条件。电镜图显示,制备第1天后粒径较小,形态较为规则、均匀,呈完整球形或椭圆形;30 d后粒径大小及形态未见明显改变。结论 采用乙醇注入-超声法制备的99Tcm-MIBI脂质体粒径较小、包封率高,且较稳定。     

Transarterial Infusion of iRGD-Modified ZrO2 Nanoparticles with Lipiodol Improves the Tissue Distribution of Doxorubicin and Its Antitumor Efficacy

PurposeTo evaluate the effect of transarterial infusion of iRGD-modified and doxorubicin-loaded zirconia-composite nanoparticles (R-DZCNs) with lipiodol in the improvement of the distribution of doxorubicin (DOX) in liver tumors and its antitumor efficacy.Materials and MethodsThe effect of R-DZCNs was evaluated in vitro by tumor cellular uptake and cytotoxicity assays. For the in vivo study, DOX distribution and antitumor efficiency were assessed. In the DOX distribution study, VX2 tumor-bearing rabbits received transarterial infusion of lipiodol with DOX, doxorubicin-loaded zirconia-composite nanoparticles (DZCNs), or R-DZCNs, respectively. DOX distribution was assessed by immunofluorescence. In the antitumor study, tumor-bearing rabbits received transarterial infusions of lipiodol with DOX, DZCNs, R-DZCNs, or saline respectively. Tumor volume was measured using magnetic resonance imaging, and the expression of apoptosis-related factors (caspase-3, Bax, Bcl-2) was analyzed by immunohistochemistry and Western blotting.ResultsR-DZCNs increased cellular uptake and caused stronger cytotoxicity. Compared with the DOX + lipiodol or DZCNs + lipiodol group, the R-DZCNs + lipiodol group showed more DOX fluorescence spots (2,449.15 ± 444.14 vs. 3,464.73 ± 632.75 or 5,062.25 ± 585.62, respectively; P < .001) and longer penetration distance (117.58 ± 19.36 vs 52.64 ± 8.53 or 83.37 ± 13.76 μm, respectively; P < .001). In the antitumor study, the R-DZCNs + lipiodol group showed smaller tumor volumes than the DOX + lipiodol or DZCNs + lipiodol group (1,223.87 ± 223.58 vs. 3,695.26 ± 666.25 or 2281.06 ± 457.21 mm³, respectively; P = .005).The greatest extent of tumor cell apoptosis was observed in R-DZCNs + lipiodol group immunohistochemistry and Western blotting results.ConclusionsTransarterial infusion of R-DZCNs with lipiodol improved the distribution of DOX and enhanced its antitumor efficacy.     

Targeting EGFR-overexpressing tumor cells using Cetuximab-immunomicelles loaded with doxorubicin and superparamagnetic iron oxide

Epidermal growth factor receptor (EGFR), a cellular transmembrane receptor, plays a key role in cell proliferation and is linked to a poor prognosis in various human cancers. In this study, we constructed Cetuximab-immunomicelles in which the anti-EGFR monoclonal antibody was linked to poly(ethylene glycol)-block-poly(?-caprolactone) (PEG–PCL) nanomicelles that were loaded with doxorubicin (DOX) and superparamagnetic iron oxide (SPIO). The specific interactions between EGFR-overexpressing tumor cells (A431) and immunomicelles were observed using confocal laser scanning microscopy (CLSM) and flow cytometry. Furthermore, the capacity of transporting SPIO into tumor cells using these immunomicelles was evaluated with a 1.5 T clinical magnetic resonance imaging (MRI) scanner. It was found that the acquired MRI T2 signal intensity of A431 cells that were treated with the SPIO-loaded and antibody-functionalized micelles decreased significantly. Using the thiazolyl blue tetrazolium bromide (MTT) assay, we also demonstrated that the immunomicelles inhibited cell proliferation more effectively than their nontargeting counterparts. Our results suggest that Cetuximab-immunomicelles are a useful delivery vehicle for DOX and SPIO to EGFR-overexpressing tumor cells in vitro and that Cetuximab-immunomicelles can serve as a MRI-visible and targeted drug delivery agent for better tumor imaging and therapy.     

Radiopharmaceutical model using99mTc-MIBI to evaluate amifostine protection against doxorubicin cardiotoxicity in rats

The aim of our study was to use an in vivo radiopharmaceutical model to investigate the cytoprotective effect of amifostine against doxorubicin-induced cardiotoxicity. Male Wistar rats were randomly divided into four groups (n = 6): 1) Saline (control); 2) Doxorubicin (DOX; 10 mg/ kg(-l) intraperitoneally); 3) Amifostine (AMI; 200 mg/kg(-1) intraperitoneally); 4) Doxorubicin plus amifostine (DOX + AMI). Amifostine was injected 30 minutes before doxorubicin in Group 4. 99mTc-MIBI, 20 MBq/0.2 ml(-1), was injected through the tail vein 72 hours after the drug administration. Rats were killed and samples of myocardium were removed by dissection 60 minutes after the injection of radiopharmaceutical. Radioactivity in each organ sample was counted using a Cd(Te) detector equipped with RAD 501 single-channel analyzer. The percent radioactivity was expressed as a percentage of the injected dose per gram of tissue (%ID/g(-1)). The %ID/g(-1) activity was calculated by dividing the activity in each sample by the total activity injected and mass of each organ. 99mTc-MIBI uptake as %ID/g(-1) was 1.194 +/- 0.502 and 0.980 +/- 0.199 in the control and AMI groups, respectively. Doxorubicin administration resulted in a significant increase in %ID/ g(-1) (3.285 +/- 0.839) (p < 0.05). Amifostine administration 30 minutes before doxorubicin injection resulted a significant decrease in %ID/g(-1) (2.160 +/- 0.791) (p < 0.05) compared with doxorubicin alone. The results showed that amifostine significantly attenuated doxorubicin-induced cardiotoxicity.