HPLC法分析南方红豆杉植株中紫杉醇的分布规律

采用高效液相色谱法对野生和人工栽培南方红豆杉约20年生植株各器官、组织中紫杉醇含量进行检测分析后发现,紫杉醇在南方红豆杉植株中具有极显著向下、向根际积累分布的规律。野生和载培红豆杉,在器官水平,从上部枝叶到根部须根,紫杉醇的积累分别为 0.007 61 %~0.100 54 %和 0.015 25 %~0.061 75 %(干重,质量分数,下同);在组织水平,从形态学上部到根部,木质组织中紫杉醇积累分别为 0.010 95%~0.044 76 %和0~0.017 58 %,周皮组织中紫杉醇的积累量也具有从形态学上部到下部增高的规律,在主枝皮中的含量分别为 0.013 32 %和 0.006 77 %,在根皮中的含量分别为 0.086 33 %和 0.059 74 %。野生南方红豆杉中紫杉醇产量比人工栽培的要高。分析过程中紫杉醇在21~105 mg/L范围内线性关系良好,r²为0.999,平均加标回收率为85.78%,相对标准偏差为1.03%。     

Dual Targeting Biomimetic Liposomes for Paclitaxel/DNA Combination Cancer Treatment

Combinations of chemotherapeutic drugs with nucleic acid has shown great promise in cancer therapy. In the present study, paclitaxel (PTX) and DNA were co-loaded in the hyaluronic acid (HA) and folate (FA)-modified liposomes (HA/FA/PPD), to obtain the dual targeting biomimetic nanovector. The prepared HA/FA/PPD exhibited nanosized structure and narrow size distributions (247.4 ± 4.2 nm) with appropriate negative charge of −25.40 ± 2.7 mV. HA/FA/PD (PTX free HA/FA/PPD) showed almost no toxicity on murine malignant melanoma cell line (B16) and human hepatocellular carcinoma cell line (HepG2) (higher than 80% cell viability), demonstrating the safety of the blank nanovector. In comparison with the FA-modified PTX/DNA co-loaded liposomes (FA/PPD), HA/FA/PPD showed significant superiority in protecting the nanoparticles from aggregation in the presence of plasma and degradation by DNase I. Moreover, HA/FA/PPD could also significantly improve the transfection efficiency and cellular internalization rates on B16 cells comparing to that of FA/PPD (p < 0.05) and PPD (p < 0.01), demonstrating the great advantages of dual targeting properties. Furthermore, fluorescence microscope and flow cytometry results showed that PTX and DNA could be effectively co-delivered into the same tumor cell via HA/FA/PPD, contributing to PTX/DNA combination cancer treatment. In conclusion, the obtained HA/FA/PPD in the study could effectively target tumor cells, enhance transfection efficiency and subsequently achieve the co-delivery of PTX and DNA, displaying great potential for optimal combination therapy.     

Paclitaxel‐Loaded Polymer Nanoparticles for the Reversal of Multidrug Resistance in Breast Cancer Cells

Novel paclitaxel‐loaded polymer nanoparticles were developed for circumventing multidrug resistance (MDR) of malignant cancerous diseases, which is an unsolved clinical problem in cancer chemotherapy. In many cases, MDR is due to the intrinsic or acquired expression of an efflux pump, the P‐170 glycoprotein (P‐gp). By encapsulating paclitaxel in a water‐soluble and biocompatible synthetic polyampholyte using a solid‐state reaction the highly water‐soluble paclitaxel‐loaded nanoparticles are formed. The resulting paclitaxel nanoparticles with an average diameter of 250 nm show a significant reversal of chemoresistance in the drug‐resistant variants (MCF7/ADR, MT3/ADR) by a factor of 100 or more. The novel paclitaxel nanoparticles enter MDR breast cancer cells by adsorptive endocytosis bypassing the P‐gp, preventing the efflux of paclitaxel and thus restoring the anti‐proliferative effect of paclitaxel.     

紫杉醇口服自乳化给药系统的处方研究

通过对油相、水、乳化剂、助乳化剂的选择和配比优化,成功地将紫杉醇包埋在乳液中,由伪三元相图得到相应的微乳区域,并结合自乳化速率、粒径、Zeta电位及紫杉醇溶解性能的研究,来筛选处方,最终确立微乳的较优配方为：十四酸异丙酯：（聚氧乙烯蓖麻油：正丁醇））（3：7（4：1））．并以简便、快捷的高效液相色谱法（HPLC）测定紫杉醇在微乳中的含量．制备的紫杉醇微乳液为泛乳光的澄清透明液体,透射电镜下观察为球状液滴,平均粒径均小于50nm．     

甘草次酸修饰的聚天冬氨酸苄酯纳米粒制备及表征

用甘草次酸(GA)修饰可生物降解的聚天冬氨酸苄酯(PBLA),将其制备成纳米粒,考察其作为药物载体的可行性.首先将GA氨基化,以引发L-天冬氨酸-β-苄酯-N-羧酸酐(BLA-NCA)开环聚合,合成不同分子量的材料GA-NH-PBLA,通过1H-NMR、FT-IR对其结构进行表征,GPC显示3种材料的分子量为1 565,3 560,5 550.采用MTT法以人肝癌细胞(HepG2)为对象,评价其作为药物载体的安全性,结果实验表明该材料无细胞毒性.采用乳化-溶剂挥发法制备包载紫杉醇(PTX)的纳米粒,并考察了不同分子量材料GA-NH-PBLA对纳米粒粒径,包封率及载药量的影响.结果显示:分子量为5 550的材料,制得的纳米粒粒径约100 nm,粒径分布均匀,球形度良好,具有良好的缓释作用,表明分子量大的材料更适合作为药物载体.     

抗癌药物筛选的新方法--紫杉醇药效的压电细胞芯片研究初探

以人肝癌细胞株HepG2为研究对象,以压电细胞芯片技术为研究手段,进行了紫杉醇药效的压电细胞芯片实时检测;同时与传统的MTT比色方法进行对比,得到了较为一致的实验结果,即药物浓度与细胞的生长抑制率呈正相关.与传统方法相比,压电细胞芯片技术更省时,更动态,是一种体外筛选药物的好方法.     

Effects of Dihydrotanshinone I on Proliferation and Invasiveness of Paclitaxel-Resistant Anaplastic Thyroid Cancer Cells

ATC is a very rare, but extremely aggressive form of thyroid malignancy, responsible for the highest mortality rate registered for thyroid cancer. In patients without known genetic aberrations, the current treatment is still represented by palliative surgery and systemic mono- or combined chemotherapy, which is often not fully effective for the appearance of drug resistance. Comprehension of the mechanisms involved in the development of the resistance is therefore an urgent issue to suggest novel therapeutic approaches for this very aggressive malignancy. In this study, we created a model of anaplastic thyroid cancer (ATC) cells resistant to paclitaxel and investigated the characteristics of these cells by analyzing the profile of gene expression and comparing it with that of paclitaxel-sensitive original ATC cell lines. In addition, we evaluated the effects of Dihydrotanshinone I (DHT) on the viability and invasiveness of paclitaxel-resistant cells. ATC paclitaxel-resistant cells highlighted an overexpression of ABCB1 and a hyper-activation of the NF-κB compared to sensitive cells. DHT treatment resulted in a reduction of viability and clonogenic ability of resistant cells. Moreover, DHT induces a decrement of NF-κB activity in SW1736-PTX and 8505C-PTX cells. In conclusion, to the best of our knowledge, the results of the present study are the first to demonstrate the antitumor effects of DHT on ATC cells resistant to Paclitaxel in vitro.     

Formulation,optimization, hemocompatibility and pharmacokinetic evaluation of PLGA nanoparticles containing paclitaxel

Objective: Paclitaxel (PTX)-loaded polymer (Poly(lactic-co-glycolic acid), PLGA)-based nanoformulation was developed with the objective of formulating cremophor EL-free nanoformulation intended for intravenous use.

Significance: The polymeric PTX nanoparticles free from the cremophor EL will help in eliminating the shortcomings of the existing delivery system as cremophor EL causes serious allergic reactions to the subjects after intravenous use.

Methods and results: Paclitaxel-loaded nanoparticles were formulated by nanoprecipitation method. The diminutive nanoparticles (143.2?nm) with uniform size throughout (polydispersity index, 0.115) and high entrapment efficiency (95.34%) were obtained by employing the Box–Behnken design for the optimization of the formulation with the aid of desirability approach-based numerical optimization technique. Optimized levels for each factor viz. polymer concentration (X1), amount of organic solvent (X2), and surfactant concentration (X3) were 0.23%, 5?ml %, and 1.13%, respectively. The results of the hemocompatibility studies confirmed the safety of PLGA-based nanoparticles for intravenous administration. Pharmacokinetic evaluations confirmed the longer retention of PTX in systemic circulation.

Conclusion: In a nutshell, the developed polymeric nanoparticle formulation of PTX precludes the inadequacy of existing PTX formulation and can be considered as superior alternative carrier system of the same.     

Preparation,characterization and in vitro anticancer activity of paclitaxel conjugated magnetic nanoparticles

In this study, magnetic nanoparticles (MNPs) coated with L-aspartic acid (F-Asp NPs) were synthesized through a co-precipitation method and conjugated with paclitaxel (PTX) (F-Asp-PTX NPs) by esterification reaction between the carboxylic acid end groups on MNPs surface and the hydroxyl groups of the PTX and studied its cytotoxic effect in vitro. The successful conjugating of PTX onto the nanoparticles (NPs) was confirmed by X-ray diffraction (XRD), thermogravimetric analysis (TGA), differential scanning calorimetry (DSC), fourier transform infrared spectroscopy (FTIR), vibrating sample magnetometer (VSM) and transmission electron microscopy (TEM) techniques. The results showed that the average size was 46.11?±?7.8 (mean?±?SD (n?=?25)) nm. The cytotoxicity of void of PTX and F-Asp-PTX NPs were compared to each other by MTT assay of the treated MCF-7 cell line. The F-Asp-PTX NPs showed pH-dependent drug release behavior. These studies specify that F-Asp-PTX NPs have a very remarkable anticancer effect, for breast cancer cell line.     

镁合金支架材料AZ81表面PLGA(PTX)/PLLA/MAO涂层的研究

在AZ81镁合金表面制备了抗腐蚀和药物释放复合涂层,并对其耐蚀性、药物释放性能及生物相容性进行研究。DSC和红外光谱数据表明,紫杉醇均匀分散在PLGA体系中。药物释放数据表明30 d后,药物释放率在80%左右。SEM、电化学阻抗、极化及镁离子释放数据表明,PLLA涂层通过有效填充微弧氧化膜表面的微孔与裂缝提高了镁合金的耐腐蚀性。血液和细胞实验表明涂层镁合金血液相容性良好,无细胞毒性。