Trans-ferulic acid-4-β-glucoside (FAG) is a monomer extracted from Radix Aconiti Lateralis Preparata, which is a potential candidate for the prevention and treatment of the cold injury. To determine the concentration FAG in rats, it is essential to develop an ultra-performance liquid chromatography coupled with tandem mass spectrometry (UPLC-MS/MS) method. The chromatographic separation was achieved by ACQUITY UPLC BEH C18 column(2.1×50mm, 1.7μm). A Xevo triple quadrupole tandem mass spectrometer was used to quantitative determination of FAG in the negative ion mode. The standard calibration curve was linear over the concentration range of 0.1-100 μg/mL and 0.0626-31.28 μg/g for rat plasma and liver tissue homogenates samples, separately. The inter-and intra-batch precision (RSD%) of the assay was ≤ 8.29% and accuracy (RE%) ranged from -7.41 to 10.99%. The matrix effect was between 92.99 and 102.39%. The oral absolute bioavailability of FAG was obtained as 1.80%. The results of tissue distribution suggested that FAG spread rarely in liver and brown adipose, which was not propitious to exert its ability to treat cold injury. In general, the above studies were significant to provide necessary information for further study.

Trans-ferulic acid-4-β-glucoside (FAG) is a monomer extracted from Radix Aconiti Lateralis Preparata, which is a potential candidate for the prevention and treatment of cold injury. To determine the concentration of FAG in rats, it is essential to develop an ultra-performance liquid chromatography coupled with MS/MS method. Chromatographic separation was achieved by an Acquity UPLC BEH C18 column (2.1 × 50 mm, 1.7 μm). A Xevo triple quadrupole tandem mass spectrometer was used to quantitatively determine FAG in the negative-ion mode. The standard calibration curve was linear over the concentration range of 0.1-100 μg/mL and 0.0626-31.28 μg/g for rat plasma and liver tissue homogenate samples, respectively. The inter- and intra-batch precision (% relative standard deviation) of the assay was ≤8.29%, and accuracy (% relative error) ranged from -7.41 to 10.99%. The matrix effect was between 92.99 and 102.39%. The oral absolute bioavailability of FAG was obtained as 1.80%. The results of tissue distribution suggested that FAG spread rarely in the liver and brown adipose, which was not propitious to exert its ability to treat cold injury. In general, these studies were significant to provide necessary information for further study.

目的 评价番茄红素固体纳米胶柬(以下简称胶束)的质量,并研究其在大鼠体内的药动学行为.方法 考察胶柬的微观结构、粒径及分布、平均载药量、包封率、人工胃液或人工肠液中的释放特性.比较番茄红素油溶液(LPO)与胶束在大鼠体内的药动学特征.结果 胶束遇水分散后形成的胶束粒径及分布都较均匀,平均载药量为1.401％±0.008％,包封率为90.26％ ±0.87％;在人工胃液中释放的程度较低,而在人工肠液中表现为持续释放.灌胃给予大鼠胶束和IPO后,以LPO为参比,胶束的相对生物利用度为183.48％.结论 将番茄红素制成胶束后,其口服生物利用度得到了一定程度的提高,改善了番茄红素因溶解度低,生物利用度低下的问题,为胶束的开发提供了参考.

The orally available novel small molecule drug ZWF is under preclinical development for an anticancer purpose. The present study aimed to assess the viability of developing ZWF as a form of oral formulation for clinical application based on the principles of biopharmaceutics and pharmacokinetics. The crucial physicochemical properties of ZWF were determined by in vitro assays. The in situ gastrointestinal absorption characteristics and in vivo pharmacokinetic behaviors of ZWF in rats were characterized. The solubility of ZWF showed a highly pH-dependent profile, decreasing from 25,392.89 to 20.48 μg/mL as the solution pH increased from 1.0 to 5.8. In PBS with a pH of 1.0 to 5.8, the LogP value of ZWF ranged from −2.35 to 2.20 and was gradually increased as the pH value increased. ZWF was partially absorbed in the stomach, and the favorable absorption sites were the duodenum, jejunum, and ileum. Pharmacokinetic studies showed that the AUC(0-t) and Cmax values of ZWF after its oral administration as a suspension prepared with 0.5% CMC-Na were increased by 18.97% and 40% than that with normal saline, providing a model oral formulation of ZWF with ideal bioavailability and system exposure in rats. From the perspective of oral absorption, ZWF possessed appealing qualities as a drug candidate and could be prepared as an oral preparation for clinical application. The present study has established a fundamental foundation for the development and quality evaluation of the ZWF oral formulations.

药物代谢动力学是定量研究药物(包括外来化学物质)在生物体内吸收、分布、排泄和代谢(简称体内过程)规律的一门学科.在动物或人体实验的基础上建立数学模型,求算相应的药物代谢动力学参数后,可以对药物在体内过程进行预测.新药和新制剂均需要进行动物或人体药物代谢动力学试验,以了解其药物代谢动力学过程. 本研究应用药物代谢动力学方法,对样品酒与参比酒进行乙醇代谢动力学研究,属于已上市产品的药物代谢动力学应用探索性研究.通过对给予既定相同剂量供试样品后机体血液乙醇浓度的经时变化特征研究,建立酒代谢动力学的试验方法,有效筛选在代谢动力学指标上具有优势的酒样,为实际产品开发和产品评估提供有效的方法.

Hupehenenine is a novel isosteroid alkaloid that was first isolated from Bulbus Hupehensis Fritillariae. The inhibitory proliferation effect of hupehenenine and its three related alkaloid derivatives, including o-caproyl-hupehenenine, o-(2-furanoyl)-hupehenenine, and Δ5(6) -isopeimine on human lung cancer cell line, human chronic myeloid leukemia cell line, and human thyroid duct cancer cell line in vitro, has been identified. This study first developed a sensitive HPLC-MS/MS method for the simultaneous quantification of hupehenenine and three alkaloid derivatives in rat plasma and tissues. The developed method was validated, and it was linear over the concentration range of 1-800 ng/mL for all analytes with R2  ≥ 0.9939 and 0.9972, respectively, in rat plasma and rat liver homogenate. The lower limit of quantitation was 1 ng/mL for all analytes. The intra-day and inter-day precision and accuracy were satisfactory. This validated method was successfully applied to investigate the pharmacokinetics and tissue distribution of hupehenenine in rats. In pharmacokinetic study, the maximum plasma concentration of rats exists gender difference. Tissue distribution study showed that hupehenenine has good affinity for multiple tissues but is unable to cross the blood-brain barrier. These results may provide a useful reference for further research of hupehenenine and its three related alkaloid derivatives.

Supersaturation drug delivery system (SDDS) based on amorphous solid dispersion (ASD) is a widely used strategy to improve oral absorption of poorly water-soluble drugs by achieving a supersaturated state where drug concentration is significantly higher than drug solubility. However, dissolved drugs tend to recrystallize in gastrointestinal (GI) tract if without effective stabilizing excipients. In this paper, well-recognized polymer (polyvinylpyrrolidone, PVP) and lipid (phosphatidylcholine, PC) excipients are combined as ASD carrier, aiming at investigating the effects on evolution of in vitro supersaturation and in vivo plasma concentration of a model poorly soluble drug indomethacin (IND). Fundamental aspects including polymer/lipid composition ratio, drug loading (DL) degree and administration dose were investigated. The in vitro dissolution profiles of ASDs were assessed by supersaturation degree, duration, maximum achievable drug concentration and dose-normalized efficiency, and correlated with in vivo pharmacokinetic data. Results showed that both in vitro and in vivo concentration-time profiles of IND were significantly varying with abovementioned factors. Solution viscosity, solid-state properties and morphology of ASDs were related to the results. This study revealed fundamental mechanisms of PVP/PC mixture effect on IND supersaturation and oral bioavailability, demonstrating that polymer/lipid mixture could be used as a promising carrier to alter supersaturation profile and oral bioavailability of SDDS products.

目的 利用旋转中心组合设计法(CCRD)优化番茄红素纳米胶束(LP-NM)的处方工艺.方法 以酪蛋白-右旋糖酐接枝共聚物(CS-g-DX)为载体,采用乳化溶剂挥发法制备LP-NM;先以载药量(DL)及胶束粒径(Dr)为初步评价指标,进行单因素筛选确定影响LP-NM制备的主要因素;再以DL、Dr及多分散系数(PDI)为评价指标,采用CCRD优化LP-NM的处方工艺,对结果进行回归方程拟合,对最优条件进行预测分析.结果 单因素筛选确定影响LP-NM制备的主要因素为酪蛋白浓度、乳化温度和载药比;各指标回归方程的拟合度较好;按处方优化得到的最佳条件制备LP-NM,各项指标的实测值与预测值接近.结论 CCRD可成功应用于LP-NM的处方工艺优化.

OBJECTIVE To investigate the in situ intestinal absorption behaviors of tectorigenin in rats.METHODS The in situ rat intestinal perfusion model was used,and the changes of tectorigenin and phenol concentration in the perfusate were determined by UV spectrophotometry and HPLC to study the absorption characteristics of tectorigenin in the whole intestine and four different intestinal segments.RESULTS Although tectorigenin was absorbed in the small intestine,it was poorly absorbed.The Ka of tectorigenin at low,medium and high concentration was 0.0997 ± 0.0177 h-1,0.1330 ± 0.0190 h-1,and 0.1513 ± 0.0134 h-1,respectively.The Ka of tectorigenin in duodenum,jejunum,ileum and colon was 0.0657 ± 0.0233 h-1,0.0576 ± 9.6 × 10-3 h-1,0.1778 ± 0.0879 h-1,and 0.0809 ± 0.0254 h-1,respectively.The Ka of tectorigenin in small intestine was in an order of ofileum ＞ colon ＞ duodenum ≈ jejunum.CONCLUSION There is a certain extent for tectorigenin absorption in the whole small intestine and the ileum is probably the main site.The mechanism of tectorigenin absorption in rat small intestine is not solely passive diffusion,there may be a carrier mediated transport.

OBJECTIVE To investigate the effect of troxerutin on cerebral ischemia-reperfusion injury in rats and to explore its possible mechanisms.METHODS SD rats were used to establish the cerebral ischemia-reperfusion models and divided into the model groups,the false operation groups,low and high dose of Troxerutin groups (25 mg· kg-1,50 mg· kg-1) for test.The behavior score,serum indexs including serum superoxide dismutase (SOD),malondialdehyde (MDA),Glutathione-S-transferase (GSH-S-transferase),nitric oxide(NO),nitric oxide synthase (NOS),xanthine oxidase (XOD) and cerebral infarct volume were observed and determined to evaluate the effects of troxerutin.RESULTS Compared with the model group,low and high dose of troxerutin could significantly decrease the level of cerebral infarction volume,behavioral score,serum MDA and SOD in a troxerutin dose-dependent manner,high dose troxerutin decrease XOD yet there was no significant effect of the low dose.The effects of troxerutin on GSH,NO and NOS had not found both in low and high doses.CONCLUSION Troxerutin has protective effect on cerebral ischemia-reperfusion injury in rats.

The purpose of this study was to establish a chemical profiling method to compare the chemical composition of herbal products by using extracts of Belamcandae Rhizoma(EBR) extracted with different polarity solvent as an example, and evaluate the quality of EBR based on the analysis of chemical profiling, efficacy evaluation and pharmacokinetics. As seen from the results of chemical profiling, the PCA and PLS-DA score plot indicated that the dots of Belamcandae Rhizoma water extracts were separated from ethanol extracts obviously, which suggested significant differences of chemical profiling existing in the different solvent extracts. The PCA and PLS-DA loading plot illustrated that the main compounds contributing to chemical profiling differences were tectoridin(TD), iristectorin B(IT B), iridin(ID), tectorigenin(TG), irigenin(IG), iristectorigein A(IG A), dichotomitin(DT) and irisflorentin(IF). Furthermore, the results of HPLC analysis demonstrated that the contents of these main compounds in ethanol extracts were significantly higher than that in water extracts (P<0.01). Both the pharmacological and hematoxylin-eosin staining studies indicated that the ethanol extracts of Belamcandae Rhizoma had a better therapeutic effect than water extracts in oral ulcer model rats (P<0.01). It is suggested that the ethanol extracts were beneficial to the absorption and bioavailability of TG which was one of the most important bioactive compounds of Belamcandae Rhizoma in pharmacokinetic study in rats. This work provided a novel method to optimize the extraction process of EBR and related herbal products. Compared with the conventional chemical fingerprint methodology, the approach proposed above is not only a powerful tool to identify efficacy related components for the quality evaluation, but also can be used to predict the therapeutic efficacy of herbal products. (C) 2018 Elsevier B.V. All rights reserved.

The purpose of this study was to characterize and evaluate tectorigenin-loaded self-microemulsifying drug delivery system (TG-SMEDDS), a previously studied preparation, and further confirm the improvement of TG in solubility and bioavailability. The appearance of TG-SMEDDS was clear and transparent, with good mobility. The microemulsion formed by TG-SMEDDS was globular, edge smooth, clear-cut, and distribution homogeneous under transmission electron microscope. The stability studies revealed that TG-SMEDDS remained stable at room temperature for at least 3 months. TG-SMEDDS showed excellent dissolution behavior that more than 90% of TG was released in only 5 min. The in situ intestinal perfusion studies indicated enhancement of absorption in four tested intestinal segments, and the main absorption site of TG was changed to duodenum. In addition, TG-SMEDDS showed significantly higher Cmax and AUC values (11-fold and 5-fold higher values, respectively; P < 0.05) than TG, and the absolute oral bioavailability of TG-SMEDDS was 56.33% (5-fold higher than that of crude TG). What’s more, the AUC0-t of crude TG and TG-SMEDDS in bile duct non-ligation rats were 6.05 and 2.80 times, respectively, than that in bile duct ligation rats, indicating the existence of enterohepatic circulation and the secretion of bile could significantly affect the absorption of TG. Further studies showed that even the bile duct was ligation, TG-SMEDDS can still keep a better oral bioavailability (179.67%, compared with crude TG in the bile duct non-ligation rats). Therefore, our study implies that SMEDDS containing TG could be an effective strategy for the oral administration of TG.

Objective To evaluate multidrug resistance of SKOV3/ADM cell for paclitaxel regulated by fangchinoline.Methods MTT tests and cell uptaking were adopted to research the activity of fangchinoline in regulating the multidrug resistance effect of paclitaxel to SKOV3/ADM.Results Cell growth inhibition test showed that paclitaxel was significantly resistant to SKOV3/ADM cells.Half inhibitory concentration (IC50) decreased to (0.542 ± 0.117),(0.924 ± 0.153)and(1.931 ± 0.375)μg·mL-1,respectively,after the combined administration of paclitaxel with 0.5,1 and 5μg·mL-1concentrations of fangchinoline.The reversal factor of resistance was 16.1,9.4 and 4.5 times,respectively,which was significantly different from that of paclitaxel alone(P<0.05).Cell uptake studies showed that,Fangchinoline could increase cell uptake pencentage of paclitaxel when the concerntration was higher than 0.5μg·mL-1(P<0.05).Conclusion Fangchinoline is a potential reversal of paclitaxel on SKOV 3 / ADM cells in multidrug resistant candidate active compounds.

OBJECTIVE To investigate the effect on liver microsomal cytochrome system in rat after changing the basic group of sodium ozagrel to tromethamine.METHODS Rats were devoted into 7 groups (n =6),including high dosage group,mid dosage group and low dosage group of sodium ozagrel and tromethamine ozagrel,and also a blank control group.The rats were treated with sodium ozagrel and tromethamine ozagrel by intravenous injection,once daily for 7 days.Then the content of liver microsomal protein and CYP450 of rat were determined by ultraviolet-visible spectrophotometer.RESULTS Protein concentration of liver microsomal of tromethamine ozagrel group was 10.14 ± 1.44 mg· mL-1,10.28 ± 1.01 mg· mL-1 and 8.10 ± 1.85 mg· mL-1,respectively.Protein concentration of liver microsomal of sodium ozagrel group was 8.26 ± 1.55 mg·mL-1,8.19 ±0.49 mg·mL-1 and 8.72 ± 1.76 mg· mL-1,respectively.The content of CYP450 of tromethamine ozagrel group was 0.537 ± 0.075 μmol· g-1,0.454-0.106 μmol· g-1 and 0.498 ± 0.071 μmol· g-1,respectively.The content of CYP450 of sodium ozagrel group was 0.504 ± 0.092 μ mol· g-1,0.450 ± 0.086 μmol· g-1 and 0.476-0.028 μmol· g-1,respectively.Compared with the control group,there were no statistically significant difference on the protein concentration of liver microsomal and the content of CYP450.CONCLUSION After changing the basic group of sodium ozagrel to tromethamine,the protein concentration of liver microsomal and the content of CYP450 in the rat by intravenous injection of tromethamine ozagrel has no statistically significant difference when compared with the sodium ozagrel group and control group.According to these,we predict that changing the basic group of sodium ozagrel to tromethamine has no significant effect on liver microsomal cytochrome system in rat.

目的 将鸢尾苷元(TG)制备成自微乳口服释药系统(SMEDDS),并对其体外溶出行为进行考察.方法 以D-optimal设计原理为基础使用Design Expert软件对制剂处方进行优化,对制备出的制剂进行质量评价及体外溶出度研究.结果 最终制备得到的TG-SMEDDS用水稀释10倍后得到粒径为(14.95±0.31) nm的O/W型微乳.粒径分布较均匀,Zeta电位为(-12.53±0.80)mV,载药量为20 mg/g,平均测定量为标示量的(99.03±0.70)％.TG-SMEDDS在pH 1.2盐酸溶液及pH6.8磷酸盐缓冲液中10 min累积溶出率均接近100％.结论 D-optimal设计可成功应用于TG-SMEDDS的处方优化,制备出的TG-SMEDDS相比TG原料药溶出度有显著改善,可以预期相较于TG原料药,TG-SMEDDS将更有利于胃肠道吸收,该研究结果可为TG剂型设计以及临床研究提供数据支持及参考.

The purpose of this study was to develop and evaluate a novel amorphous solid dispersion system for tectorigenin (TG). TG is one of isoflavone aglycones extracted from Iris tectorum and flowers of Pueraria thunbergiana, but its poor water solubility and low membrane permeability have severely restricted the clinical application. To increase the aqueous solubility and oral bioavailability of TG, we prepared the solid dispersions of tectorigenin (TG-SD) using a simple solvent evaporation process with TG, polyvinylpyrrolidone (PVP) and PEG4000 at weight ratio of 7:54:9 after tested in several ratios. The prepared solid dispersions of tectorigenin are duly characterized for drug morphological conversion, in vitro dissolution and in vivo bioavailability. The X-ray diffraction (XRD), differential scanning calorimetry (DSC) and scanning electron microscopy (SEM) studies have indicated the morphological conversion of tectorigenin to amorphous form. In vitro release profiles revealed that the % release of TG-SD was achieved 4.35-fold higher than that of the pure drug after 150 min. The oral bioavailability of the solid dispersion in rats was also increased based on AUC0–t and C max of TG-SD, which were 4.8- and 13.1-fold higher than that of TG crystal, respectively. It is worth noting that physical mixture containing TG, PEG4000 and PVP produced a similar level of oral exposure as TG-SD, suggesting that PEG4000 and PVP were able to enhance bioavailability of TG in rats. However, with the reduction of particle size, TG-SD provided the fastest oral absorption compared to physical mixture and pure drug. These results demonstrated that the efficacy of solid dispersions for the enhancement of TG oral bioavailability was by increasing its aqueous solubility and the solid dispersion formulation could be a viable option for enhancing the oral bioavailability of TG.

目的 用LC-MS/MS法测定人血清中的25-羟基维生素D3.方法 血清样品采用固相萃取后浓缩处理;采用C18色谱柱,流动相为甲醇-0.5％甲酸水(90∶10),流速0.3 mL· min-1,柱温30℃;质谱使用电喷雾离子化(ESI),正离子模式,多反应监测(MRM),25-羟基维生素D3检测离子质荷比为m/z 401.3→159.2,羟基喜树碱内标的为m/z 365.1→321.3.结果 0.633 ～ 633 ng· mL-1 125-羟基维生素D3与峰面积呈良好的线性关系(r=0.9999),最低检测浓度为0.633 ng·mL-1,准确度及精密度均符合生物样品的分析要求.结论 所用方法专属性强,适用于25-羟基维生素D3血清样品的定量分析.

目的 研究他克莫司软膏对离体巴马香猪皮的透皮吸收情况,并比较他克莫司软膏的供试制剂与参比制剂的体外透皮渗透量和渗透速率.方法 采用改良Franz扩散池法,用反相HPLC法定量测定接受室溶液中他克莫司浓度及皮内滞留药物.结果 他克莫司经离体巴马香猪皮的透过率随时间的延长而增加;48 h内,他克莫司软膏的供试制剂与参比制剂经离体巴马香猪皮的累积透过量分别为22.87±1.31、21.61±2.26μg·cm-2,平均渗透速率分别为0.7669、0.7149 μg·cm-2·h-1.结论 在透皮吸收研究中,他克莫司软膏的供试制剂与参比制剂的透皮渗透程度和渗透速率差异无统计学意义.

目的 研究鸢尾苷元在小鼠体内的组织分布,并探讨与中药归经理论的关系.方法 用HPLC法测定静脉注射给药后不同时间内小鼠各组织中鸢尾苷元的浓度,流动相为水(磷酸调pH3)-甲醇(42∶58),流速1.0 mL·min-1,柱温35℃,检测波长268 nm.结果 鸢尾苷元的各组织匀浆样品在一定浓度范围内线性关系良好,精密度和准确度均符合生物样品测定的要求;小鼠静注2 mg·kg-1鸢尾苷元后,药物在小鼠的心、肝、脾、肾、睾丸、小肠、骨骼肌组织内均有分布,其中以小肠与肝脏组织中分布较多,而肺、脑组织中未检测到药物.结论 鸢尾苷元在各组织器官的分布有显著差异,对评价射干及鸢尾苷元的中医归经特点具有重要意义.