熔融高速搅拌法制备双氯芬酸钠缓释胶囊的研究

目的 研究熔融高速搅拌法制备双氯芬酸钠缓释胶囊及其体外释药行为。方法 采用KJZ-10型熔融高速搅拌制粒机制备含药微丸,以微丸体外释放度为指标,考察处方工艺因素对微丸体外释放度的影响,通过释放度曲线药动力学拟合确定微丸缓释机制。结果 双氯芬酸钠微丸体外释放行为符合Higuchi方程 Y=35.43t_1/2-16.152 3(r=0.998 8),释药机制主要是骨架溶蚀和扩散释放。结论 该技术制备的双氯芬酸钠缓释微丸具有较好的释药性能及良好的缓释效果。     

双氯芬酸钠缓释微丸胶囊的制备

目的：制备膜控释双氯芬酸钠缓释微丸胶囊;方法：以乙基纤维素为包衣材料,以PEG100为致孔剂,经薄膜包衣制备双氯芬酸钠缓释微丸胶囊,并以体外释放度试验来评价;结果：制备的双氯芬酸钠缓释微丸胶囊的体外释药行为符合零级方程,批与批之间的重现性良好;结论：采用该法制备的双氯芬酸钠缓释微丸胶囊缓释效果好,制备工艺稳定。     

熔融法制备5-单硝酸异山梨酯缓释微丸及其体外释放考察

目的制备5-单硝酸异山梨酯缓释微丸,并对其体外释放度进行考察。方法采用高速熔融搅拌法制备5-单硝酸异山梨酯微丸,以微丸体外溶出参数t50为考察指标设计正交试验对处方进行筛选,通过释放度曲线药物代谢动力学拟合确定微丸的缓释机制。结果确定的最佳处方为每粒胶囊含有主药60 mg、预胶化淀粉16 mg、固体石蜡100 mg、单硬脂酸甘油酯16 mg、磷酸氢钙120 mg、硫酸钙适量;所制备的5-单硝酸异山梨酯缓释微丸的体外释放行为符合Higuchi方程Qt=0.242 2t1/2+0.084 2(r=0.996 0)。结论采用高速熔融搅拌法制备的5-单硝酸异山梨酯缓释微丸表面光滑,圆整度高,具有良好的缓释效果。     

双氯芬酸钠壳聚糖明胶复合微球的释药机制研究

目的制备双氯芬酸钠壳聚糖明胶复合微球(DS-CGMs),探讨其体外释药机制。方法建立DS-CGMs体外释放度的测定方法;研究药物从微球中释放的影响因素;分别考查双氯芬酸钠明胶微球(DS-GMs)、双氯芬酸钠壳聚糖微球(DS-CMs)及DS-CGMs的体外释药情况,并通过数学原理和相关模型探讨其释放行为和机制。结果DS-GMs、DS-CMs以扩散和骨架溶蚀作用释放,而DS-CGMs呈骨架溶蚀作用机制,具有更好的缓释效果。结论获得较为满意的DS-CGMs,其体外释药具有一定的缓释性。     

盐酸美金刚缓释微丸的制备及体外释放度考察

目的:制备盐酸美金刚缓释微丸,并对其体外释放度进行考察。方法:采用流化床包衣法制备盐酸美金刚载药微丸,再用Eudragit RL 30D和Eudragit RS 30D进行包衣,制成盐酸美金刚缓释微丸,并考察盐酸美金刚缓释微丸的体外释药行为。结果:体外释放度试验显示,制备的盐酸美金刚缓释微丸在24 h内平稳释放且释药完全,释药规律符合零级释药模型。结论:用本方法制备的盐酸美金刚缓释微丸具有缓释效果。     

双氯芬酸钠缓释微丸的制备及体外释放度

目的 :制备双氯芬酸钠缓释微丸 (DS SRMP)并研究其体外释药性能。方法 :选用乙基纤维素等辅料为阻滞剂制备了DS SRMP ,并采用紫外分光光度法进行了体外释放度的研究。结果 :实验研究表明DS SRMP体外释药符合零级动力学过程 ,Kr0 =8.78%·h-1(t≤ 8h)。结论 :DS SRMP具有较好的释药性能 ,为一种较理想的口服缓释剂型。     

甲磺酸双氢麦角毒碱缓释微丸的研制

目的：制备甲磺酸双氢麦角毒碱缓释微丸,并研究其体外释药情况。方法：采用离心造粒法制备甲磺酸双氢麦角毒碱素丸,在流化床内采用丙烯酸树脂水分散体对其包衣,制备甲磺酸双氢麦角毒碱缓释微丸。用释放度测定法考察影响药物释放的各种因素,研究包衣微丸体外释药机制。结果：所制微丸体外释药过程基本符合Higuchi方程：Y=0．41＋29．22t1/2（r=0．9782）。结论：甲磺酸双氢麦角毒碱缓释微丸体外释药缓慢、平稳。     

法罗培南钠缓释微丸的制备及体外释放特性考察

目的制备法罗培南钠缓释微丸,并对其体外释药特性进行研究。方法挤出滚圆法制备载药丸芯;以乙基纤维素N-100为包衣材料,以聚维酮k30为致孔剂,流化床包衣制备法罗培南钠缓释微丸;考察包衣增重、致孔剂用量等因素对缓释微丸释放度的影响,并考察其体外释药特征及释药机制。结果所制备的微丸有明显的缓释特征,体外释放符合Higuchi释药动力学方程。结论制备的法罗培南钠缓释微丸具有较理想的体外缓释效果。     

复方盐酸伪麻黄碱缓释微丸的制备及其释放度考察

目的制备含萘普生钠和盐酸伪麻黄碱的复方缓释微丸,考察理论聚合物包覆量、抗黏剂用量、热处理时间等因素对盐酸伪麻黄碱缓释微丸释放度的影响。方法采用挤出滚圆法制备萘普生钠速释微丸及盐酸伪麻黄碱含药丸芯,采用丙烯酸树脂水分散体(EudragitRS 30D)流化床包衣技术制备盐酸伪麻黄碱缓释微丸,采用紫外分光光度法测定萘普生钠的含量及溶出度,采用HPLC法测定盐酸伪麻黄碱的含量及释放度。结果萘普生钠、微晶纤维素、硫酸钠质量比为10∶9∶1时,制得的萘普生钠速释微丸圆整度好,脆碎度低,10 min累积释药大于90%;当盐酸伪麻黄碱缓释微丸的理论聚合物包覆量为50%、滑石粉用量为聚合物质量的40%、热处理时间为24 h时,药物释放过程接近零级释药模型。结论所制备的萘普生钠速释微丸及盐酸伪麻黄碱缓释微丸体外释药均符合速释和缓释要求,后者的体外释药过程接近零级释药模型。     

丹参总酚酸胃漂浮缓释微丸的制备及其体外释放度考察

目的制备丹参总酚酸胃漂浮缓释微丸,初步评价其体外释药特性及释药机制。方法用挤出滚圆法制备素丸,以乙基纤维素为包衣材料,流化床制备丹参总酚酸缓释微丸,并考察其体外释放度。结果包衣微丸表面光滑圆整,不同批次间微丸释药重现性良好。在人工胃液中,微丸的8 h漂浮率达到95%左右,微丸的体外释药符合Higuchi方程。结论丹参总酚酸缓释微丸具有较理想的体外漂浮及缓释效果。     

Andrographolide suppresses endothelial cell apoptosis via activation of phosphatidyl inositol-3-kinase/Akt pathway

Andrographolide (Andro), an active component isolated from the Chinese official herbal Andrographis paniculata, which has been reported to prevent oxygen radical production and thus prevent inflammatory diseases. In this study, we investigated the molecular mechanisms and signaling pathways by which Andro protects human umbilical vein endothelial cells (HUVECs) from growth factor (GF) deprivation-induced apoptosis. Results demonstrated that HUVECs undergo apoptosis after 18 hr of GF deprivation but that this cell death was suppressed by the addition of Andro in a concentration-dependent manner (1-100 microM). Andro suppresses the mitochondrial pathway of apoptosis by inhibiting release of cytochrome c into the cytoplasm and dissipation of mitochondrial potential (Deltapsi(m)), as a consequence, prevented caspase-3 and -9 activation. Treatment of endothelial cells with Andro-induced activation of the protein kinase Akt, an anti-apoptotic signal, and phosphorylation of BAD, a down-stream target of Akt. Suppression of Akt activity by wortmannin, by LY-294002 and by using a dominant negative Akt mutant abolished the anti-apoptotic effect of Andro. In contrast, the ERK1/2 activities were not affected by Andro. The ERK1/2 inhibitor, PD98059 failed to antagonize the protective effect of Andro. In conclusion, Andro exerts its anti-apoptotic potential via activation of the Akt-BAD pathway in HUVECs and thus may represent a candidate of therapeutic agent for atherosclerosis.     

穿心莲内酯对人肺腺癌A549细胞NF-κB通路的调控作用

目的探讨穿心莲内酯对肿瘤细胞生长的作用及机制。方法人肺腺癌A549细胞分别与穿心莲内酯1.5～30μmol·L-1作用24h,以及穿心莲内酯30μmol·L-1作用4～24h。用MTT法检测A549细胞存活率;Western印迹法检测在肿瘤坏死因子α(TNF-α)10μg·L-1刺激下,穿心莲内酯30μmol·L-1对NF-κB信号通路中的相关蛋白NF-κB抑制因子α(IκBα)、磷酸化IκBα、IκB激酶β(IKKβ)和磷酸化IKKβ表达的影响;ELISA法检测穿心莲内酯对A549细胞核内NF-κBDNA结合活性的影响。结果穿心莲内酯的浓度和作用时间与A549细胞的存活率密切相关,穿心莲内酯30μmo·lL-1作用24h,A549细胞的存活率下降到(22.0±1.2)%,而穿心莲内酯1.5μmol·L-1作用24h或者穿心莲内酯30μmol·L-1作用4h对A549细胞的存活率几乎无影响。Western印迹法显示,穿心莲内酯能够抑制TNF-α诱导的NF-κB信号通路中IKKβ的磷酸化,抑制IκBα的磷酸化,推迟IκBα的降解,对IKKβ的表达无影响。穿心莲内酯还能够抑制TNF-α诱导的A549细胞核内NF-κBp65蛋白的DNA结合活性,抑制率达32%。结论穿心莲内酯通过影响NF-κB信号通路抑制A549细胞的生长。     

Andrographolide down-regulates hypoxia-inducible factor-1α in human non-small cell lung cancer A549 cells

Andrographolide (Andro), a diterpenoid lactone isolated from a traditional herbal medicine Andrographis paniculata, is known to possess multiple pharmacological activities. In our previous study, Andro had been shown to inhibit non-small cell lung cancer (NSCLC) A549 cell migration and invasion via down-regulation of phosphatidylinositol 3-kinase (PI3K)/Akt signaling pathway. Here we demonstrated that Andro inhibited the expression of hypoxia-inducible factor-1α (HIF-1α) in A549 cells. HIF-1α plays an important role in tumor growth, angiogenesis and lymph node metastasis of NSCLC. The Andro-induced decrease of cellular protein level of HIF-1α was correlated with a rapid ubiquitin-dependent degradation of HIF-1α, and was accompanied by increased expressions of hydroxyl-HIF-1α and prolyl hydroxylase (PHD2), and a later decrease of vascular endothelial growth factor (VEGF) upon the treatment of Andro. The Andro-inhibited VEGF expression appeared to be a consequence of HIF-1α inactivation, because its DNA binding activity was suppressed by Andro. Molecular data showed that all these effects of Andro might be mediated via TGFβ1/PHD2/HIF-1α pathway, as demonstrated by the transfection of TGFβ1 overexpression vector and PHD2 siRNA, and the usage of a pharmacological MG132 inhibitor. Furthermore, we elucidated the involvement of Andro in HIF-1α transduced VEGF expression in A549 cells and other NSCLC cell lines. In conclusion, these results highlighted the potential effects of Andro, which may be developed as a chemotherapeutic or an anti-angiogenesis agent for NSCLC in the future.     

Andrographolide inhibits human hepatoma-derived Hep3B cell growth through the activation of c-Jun N-terminal kinase

Andrographolide (Andro) is a potentially anti-inflammatory diterpenoid lactone isolated from the traditional herbal medicine ANDROGRAPHIS PANICULATA, which has been effectively used for the treatment of infection, inflammation, cold, fever and diarrhea in China for centuries. In the current study, we found that Andro significantly decreased the number of surviving hepatoma-derived Hep3B cells in the MTT assay and induced cell apoptosis. Further study showed that Andro induced activation of mitogen-activated protein kinases (MAPKs) including p38 kinase, c-Jun N-terminal kinase (JNK) and extracellular signal-related kinases (ERK1/2), but had no significant effect on caspase-3, Bcl-xL and Bcl-2, which are apoptosis-related proteins. Moreover, inhibition of JNK activation partially rescued the toxic effect of Andro on Hep3B cells. Therefore, our results indicate that the JNK signaling pathway plays an important role in the toxic effect of Andro on Hep3B cells.     

Synergistic anticancer effects of andrographolide and paclitaxel against A549 NSCLC cells

Context: Paclitaxel (PTX) is widely used in chemotherapy for cancer treatment; however, it has some serious side effects. Andrographolide (Andro) is a potential cancer therapeutic agent isolated from Andrographis paniculata (Burm. f.) Nees (Acanthaceae).

Objective: The objective of this study is to evaluate the effects of PTX combined with Andro against A549 cells.

Materials and methods: The effects of 24–48?h treatment with 0.48–60.75?nM PTX and 5.10–328.0?μM Andro on cellular proliferation, apoptosis, cell cycle and intracellular reactive oxygen species (ROS) were determined by sulphorhodamine B assay, Annexin V-FITC/PI apoptosis detection, PI staining and ROS assay, respectively. Synergy was determined using combination index. The antitumour efficacy of 20?mg/kg PTX with 100?mg/kg Andro was studied in a xenograft murine model.

Results: IC₅₀ value of the PTX combined with Andro against A549 cells was 0.5–7.4?nM, which was significantly lower than that of PTX (15.9?nM). PTX with 10?μM Andro caused (1.22–1.27)-fold apoptosis and 1.7-fold ROS accumulation compared with PTX alone. N-Acetylcysteine, a ROS scavenger, blocked this synergy in vitro. In contrast, G2/M phase cell cycle arrest resulting from PTX was not potentiated by Andro. Moreover, PTX in combination with Andro inhibited the growth of A549 transplanted tumours by 98%.

Discussion and conclusion: The results indicate that the combination of PTX and Andro exert significant synergistic anticancer effect on A549 cells in vitro and in vivo. The synergy may be the result of the accumulation of ROS. The combination of Andro and PTX represents a potential strategy for the treatment of A549 cells.     

Cellular glutathione content modulates the effect of andrographolide on β-naphthoflavone-induced CYP1A1 mRNA expression in mouse hepatocytes

We previously reported that andrographolide (Andro), a major bioactive constituent of Andrographis paniculata, synergistically enhanced the inducible expression of CYP1A1 mRNA. In this study, although the synergism was confirmed at 24h after the start of treatment with Andro and β-naphthoflavone (βNF), a CYP1A inducer, the expression was profoundly suppressed at an earlier phase, namely at 6-12h, when the βNF-induced expression peaked. Although oxidized glutathione (GSSG) levels were higher in co-treated cells at 6 and 24h, levels of reactive oxygen species varied depending on the treatment period and species, indicating no relation to the synergistic expression of CYP1A1 mRNA. Glutathione (GSH) and N-acetyl-l-cysteine (NAC) significantly enhanced the βNF-induced expression, and partly reversed the suppressive effect of Andro in the early phase. At 24h, the addition of GSH or NAC had no effect on βNF-induced CYP1A1 mRNA expression, but significantly reduced the synergistic effect of Andro. The synergistic effect was enhanced by l-buthionine-(S,R)-sulfoximine, a GSH depleter. Furthermore, H(2)O(2) and ascorbic acid further modified the profile of synergism of Andro on βNF-inducible CYP1A1 mRNA expression. These results suggest that GSH status might be involved in βNF-induced CYP1A1 mRNA expression, and the interaction of Andro with GSH might modulate the expression.     

Critical role of pro-apoptotic Bcl-2 family members in andrographolide-induced apoptosis in human cancer cells

Andrographolide (Andro), a diterpenoid lactone isolated from a traditional herbal medicine Andrographis paniculata, is known to possess potent anti-inflammatory activity. In this study, Andro induced apoptosis in human cancer cells via activation of caspase 8 in the extrinsic death receptor pathway and subsequently with the participation of mitochondria. Andro triggered a caspase 8-dependent Bid cleavage, followed by a series of sequential events including Bax conformational change and mitochondrial translocation, cytochrome c release from mitochondria, and activation of caspase 9 and 3. Inhibition of caspase 8 blocked Bid cleavage and Bax conformational change. Consistently, knockdown of Bid protein using small interfering RNA (siRNA) technique suppressed Andro-induced Bax conformational change and apoptosis. In conclusion, the pro-apoptotic Bcl-2 family members (Bid and Bax) are the key mediators in relaying the cell death signaling initiated by Andro from caspase 8 to mitochondria and then to downstream effector caspases, and eventually leading to apoptotic cell death.     

Carbon isotope ratio mass spectrometry for detection of endogenous steroid use: A testing strategy

Isotope ratio mass spectrometry (IRMS) testing is performed to determine if an atypical steroid profile is due to administration of an endogenous steroid. Androsterone (Andro) and etiocholanolone (Etio), and/or the androstanediols (5α‐ and 5β‐androstane‐3α,17β‐diol) are typically analyzed by IRMS to determine the ¹³C/¹²C ratio. The ratios of these target compounds are compared to the ¹³C/¹²C ratio of an endogenous reference compound (ERC) such as 5β‐pregnane‐3α,20α‐diol (Pdiol). Concentrations of Andro and Etio are high so ¹³C/¹²C ratios can easily be measured in most urine samples. Despite the potentially improved sensitivity of the androstanediols for detecting the use of some testosterone formulations, additional processing steps are often required that increase labour costs and turnaround times. Since this can be problematic when performing large numbers of IRMS measurements, we established thresholds for Andro and Etio that can be used to determine the need for additional androstanediol testing. Using these criteria, 105 out of 2639 urine samples exceeded the Andro and/or Etio thresholds, with 52 of these samples being positive based on Andro and Etio IRMS testing alone. The remaining 53 urine samples had androstanediol IRMS testing performed and 3 samples were positive based on the androstanediol results. A similar strategy was used to establish a threshold for Pdiol to identify athletes with relatively ¹³C‐depleted values so that an alternative ERC can be used to confirm or establish a true endogenous reference value. Adoption of a similar strategy by other laboratories can significantly reduce IRMS sample processing and analysis times, thereby increasing testing capacity. Copyright © 2013 John Wiley & Sons, Ltd.     

Andrographolide relieved pathological pain generated by spared nerve injury model in mice

Context: Andrographolide (Andro), found in large quantities in Andrographis paniculata Nees (Acanthaceae), is anti-inflammatory, especially in the central nervous system (CNS) glia.

Objective: The objective of this study is to test Andro’s ability to reduce allodynia in a spared nerve injury model.

Material and methods: Male 30?g BalbC mice were divided into four groups: (1) Sham-operated control (Sham-group); (2) nerve injured and treated with saline (Saline-group); (3) nerve injured and treated with Andro (Andro-group); (4) nerve injured and treated with non-steroidal anti-inflammatory drugs (NSAIDS) (NSAIDS-group). Andro or NSAIDS (diclofenac salt) were injected intraperitoneally at 5?mg/kg body weight daily. Mechanical allodynia was assessed by von Frey tests at 3, 7, and 14?d. For immunohistochemical analysis, samples were collected at 7?d.

Results: The threshold for inducing allodynia increased and the response percentage reduced in the Andro-group when compared with the Saline-group, as well as when compared with NSAIDS groups throughout 3–14?d. The ratio of threshold for OP-Andro/OP-saline and for OP-Andro/OP-NSAIDS groups was 20.42 and 11.67 at 14?d, respectively. The ratio of response percentage for OP-Andro/OP-saline and for OP-Andro/OP-NSAIDS was 0.32 and 0.39 at 14?d, respectively. Interleukin-1 (IL-1) immunostaining in the spinal cord was reduced in the Andro-group. Astrocytic activities were not significantly reduced in the Andro-group compared with the Saline-group at 7?d post-operation (PO)

Conclusions: Andro reduced mechanical allodynia more than NSAIDS at the same concentration, and the observed behaviour was associated with a reduction in inflammatory cytokine produced in the spinal cord.