高效液相色谱-质谱联用法测定姜黄素在大鼠体内的分布

目的:采用高效液相色谱-质谱联用法,测定姜黄素在大鼠血浆和组织中药物浓度随时间变化的分布规律及药物代谢动力学的特点。方法:大鼠口服灌胃姜黄素后,收集血浆和组织样品,通过有机溶剂乙酸乙酯分离提取,采用高效液相色谱-质谱联用法分析,选择C18色谱分析柱,乙腈（85%）和水(5%)为流动相,检测血浆和组织中的姜黄素药物浓度。质谱条件为电喷雾电离正离子方式(ESI＋),多反应监测(MRM)模式。结果:姜黄素在血中的达峰时间为0.5 h,曲线下面积(AUC)为(236±6)ng?h/mL,峰值血药浓度为(91±5)ng/mL;心脏中的达峰时间为1 h,肝脏和肾脏中的达峰时间均为2 h。在心脏和肝脏中,曲线下面积(AUC)分别为(127±3)ng?h/mL和(166±5)ng?h/mL,均显著高于其他组织。结论:我们建立的高效液相色谱-质谱联用分析方法,可有效监测姜黄素在血中或组织器官中药物浓度随时间变化的规律,此方法简便、重复性高、灵敏度强。大鼠口服姜黄素后,可被吸收入血,组织器官分布广泛。     

Enhancement of curcumin oral absorption and pharmacokinetics of curcuminoids and curcumin metabolites in mice

Purpose  

Curcumin has shown a variety of biological activity for various human diseases including cancer in preclinical setting. Its poor oral bioavailability poses significant pharmacological barriers to its clinical application. Here, we established a practical nano-emulsion curcumin (NEC) containing up to 20% curcumin (w/w) and conducted the pharmacokinetics of curcuminoids and curcumin metabolites in mice.     

姜黄素对人鼻咽癌CNE-2Z细胞的放射增敏作用及其作用机制

目的:研究姜黄素(curcumin)对人鼻咽癌CNE-2Z细胞放射增敏作用并初步探讨其作用机制。方法:应用MTT法检测姜黄素药物毒性,用克隆形成实验观察其对放射敏感性的影响,用流式细胞仪(FCM)技术分析对CNE-2Z细胞周期分布和姜黄素联合辐射引起的细胞周期阻滞的影响。结果:不同浓度的姜黄素作用于CNE-2Z细胞后,其细胞毒性呈剂量依赖性,在较低浓度(10μmol/L)时,即可降低放射后CNE-2Z细胞的克隆形成率,其放射增敏比为1.52±0.25。姜黄素以及姜黄素联合辐射作用CNE-2Z细胞24h后,细胞周期主要阻滞在辐射敏感时相G2/M期。结论:姜黄素对CNE-2Z细胞有放射增敏作用,其机制可能与其引起的细胞周期阻滞等因素有关。     

Antitumour effects of metformin and curcumin in human papillomavirus positive and negative head and neck cancer cells

The incidence of oropharyngeal squamous cell carcinoma (OPSCC) has significantly increased in recent decades due to human papillomavirus (HPV)‐mediated oncogenesis. Unfortunately, a growing number of HPV‐positive (+) OPSCC survivors are living with the irreversible side effects of treatment. The novel, well‐tolerated chemotherapeutics with improved side effect profiles are, therefore, in high demand. Metformin is one such drug, widely used as a first‐line oral agent in the treatment of type 2 diabetes mellitus. Curcumin is another well‐tolerated agent quickly gaining attention for its medicinal properties. Both metformin and curcumin have been shown to display anticancer properties. This study aimed to determine the antitumor effects of these agents, individually and combined, in HPV+???? ???and HPV‐negative (?) head and neck squamous cell carcinoma (HNSCC) cell lines. This was achieved by assessing the efficacy of varying drug concentrations on the overall cell viability, proliferation, and expression of common HNSCC biomarkers. The results from protein and RNA expression data are highly variable, as expected, with multiple pathways being affected in cancer. 3‐(4,5‐Dimethylthiazol‐2‐yl)‐2,5‐diphenyltetrazolium bromide assays and immunofluorescence microscopy suggest that both agents are capable of slowing proliferation and inducing apoptosis. We conclude that curcumin and metformin display effective antitumor effects in both HPV+ and HPV? HNSCC cell lines. The curcumin effects appear more pronounced in the HPV? cell lines. Metformin appears to be more effective at reducing the overall cell numbers in HPV+ cell lines. Metformin and curcumin combined did not appear to have synergistic effects on the proliferation or apoptosis of the treated cell lines.     

姜黄素对子宫颈癌HeLa细胞增殖抑制作用及其机制的研究

目的:探讨姜黄素(Curcum in)对体外培养人子宫颈癌HeLa细胞抗癌作用及分子机制。方法:MTT法检测细胞增殖,流式细胞仪检测凋亡和细胞周期,PI/Hoechst33258荧光双染法检测细胞凋亡,W estern b lot法检测细胞凋亡相关蛋白Bc l-2和Bax蛋白的表达。结果:姜黄素对HeLa细胞生长有抑制作用,并呈剂量依赖性;流式细胞仪分析证实姜黄素能使HeLa细胞阻滞在S期,并出现亚二倍体凋亡峰;荧光双染法可见凋亡细胞;W estern b lot结果显示Bax蛋白表达均上调,而Bc l-2的表达无明显影响。结论:姜黄素对人子宫颈癌HeLa细胞的增殖具有显著的抑制作用并可诱导细胞凋亡;Bax蛋白的表达上调可能参与了诱导细胞凋亡。     

A novel self-microemulsifying formulation of paclitaxel for oral administration to patients with advanced cancer

To explore the parmacokinetics, safety and tolerability of paclitaxel after oral administration of SMEOF#3, a novel Self-Microemulsifying Oily Formulation, in combination with cyclosporin A (CsA) in patients with advanced cancer. Seven patients were enrolled and randomly assigned to receive oral paclitaxel (SMEOF#3) 160 mg+CsA 700 mg on day 1, followed by oral paclitaxel (Taxol) 160 mg+CsA 700 mg on day 8 (group I) or vice versa (group II). Patients received paclitaxel (Taxol) 160 mg as 3-h infusion on day 15. The median (range) area under the plasma concentration-time curve of paclitaxel was 2.06 (1.15-3.47) microg h ml(-1) and 1.97 (0.58-3.22) microg h ml(-1) after oral administration of SMEOF#3 and Taxol, respectively, and 4.69 (3.90-6.09) microg h ml(-1) after intravenous Taxol. Oral SMEOF#3 resulted in a lower median T(max) of 2.0 (0.5-2.0) h than orally applied Taxol (T(max)=4.0 (0.8-6.1) h, P=0.02). The median apparent bioavailability of paclitaxel was 40 (19-83)% and 55 (9-70)% for the oral SMEOF#3 and oral Taxol formulation, respectively. Oral paclitaxel administered as SMEOF#3 or Taxol was safe and well tolerated by the patients. Remarkably, the SMEOF#3 formulation resulted in a significantly lower T(max) than orally applied Taxol, probably due to the excipients in the SMEOF#3 formulation resulting in a higher absorption rate of paclitaxel.     

姜黄素碘混肝动脉给药对兔VX2肝移植瘤生长转移的影响

目的:在动物肝癌模型中初步评价姜黄素碘混肝动脉给药对肿瘤生长转移的影响。方法:54只新西兰兔随机分成对照组和实验组,将VX2瘤种植在兔肝左叶,2周后开腹直视下穿刺肝动脉,分别经肝动脉缓慢推注碘油、姜黄素碘油乳剂,2周后观察肿瘤的生长情况和肝内、双肺及腹腔淋巴结等转移情况,定性检测基质蛋白酶-2(MMP-2)和MMP-9。结果:54只兔成功种植肝移植瘤52只(96.23%)。52只兔肝动脉穿刺成功48只(92.31%)。对照组肿块长径平均约为2.5cm,实验组约为2.3cm。2组肿瘤生长率差异无统计学意义,t=1.73,P=0.38;2组兔肝内转移发生率均在50%左右,χ2=0.20,P=0.92;2组兔的肺及腹腔淋巴结转移数目均分别与MMP-2、MMP-9阳性细胞数呈正相关。对照组的肺及腹腔淋巴结转移总数目、MMP-2、MMP-9阳性细胞数均明显高于实验组,P值分别为0、0.005、0和0。结论:姜黄素在活体动物模型上具有抗侵袭与转移的能力。     

Pharmacokinetics ofcis-dichloro-trans-dihydroxy-bis-isopropylamine platinum IV (iproplatinum,chip) in patients with normal and impaired renal function and following intraperitoneal administration

CHIP is a quadrivalent platinum (Pt) complex, introduced clinically as a less toxic alternative tocis-platinum. The drug’s major route of excretion is via the kidneys, and in this study the pharmacokinetics of unchanged CHIP, filterable Pt and total Pt have been determined following intravenous administration to patients with a range of renal function. Total Pt and filterable Pt in plasma decayed biexponentially and was fitted to a two-compartment model, whereas unchanged CHIP declined monoexponentially and was best fitted to a one-compartment model, according to Akaike’s information criteria. There is a correlation between the unchanged CHIP clearance and⁵¹Cr-EDTA clearance. The pharmacokinetics of CHIP was determined following intraperitoneal (i.p.) administration (dose, 150–300 mg m⁻² 4 h dwell time) and a regional advantage (peritoneal peak concentration/plasma peak concentration) of approximately 30 fold was seen. It is likely that the dose of CHIP will need to be reduced in patients with impaired renal function, and the use of i.p. CHIP in ovarian carcinoma warrants further study.     

甲状腺肿瘤患者外周血和肿瘤组织中褪黑素的检测及其意义

目的: 检测甲状腺肿瘤患者外周血和肿瘤组织褪黑素（melatonin,MLT）的水平，探讨其临床意义。方法：选取2002年2月至2003年3月上海长征医院和第411医院成人行甲状腺肿瘤手术切除标本30例，同时设甲状腺非肿瘤疾病20例和健康体检者20例为对照组。用ELISA法检测30例甲状腺肿瘤     

Cyclohexanone curcumin analogs inhibit the progression of castration‐resistant prostate cancer in vitro and in vivo

Many prostate cancer patients develop resistance to treatment called castration‐resistant prostate cancer (CRPC) which is the major cause of recurrence and death. In the present study, four cyclohexanone curcumin analogs were synthesized. Additionally, their anticancer progression activity on CRPC cell lines, PC3 and PLS10 cells, was examined. We first determined their anti‐metastasis properties and found that 2,6‐bis‐(4‐hydroxy‐3‐methoxy‐benzylidene)‐cyclohexanone (2A) and 2,6‐bis‐(3,4‐dihydroxy‐benzylidene)‐cyclohexanone (2F) showed higher anti‐invasion properties against CRPC cells than curcumin. Analog 2A inhibited both MMP‐2 and MMP‐9 secretions and activities, whereas analog 2F reduced only MMP activities. These findings suggest that the compounds may inhibit CRPC cell metastasis by decreased extracellular matrix degradation. Analog 2A, the most potent analog, was then subjected to an in vivo study. Similar to curcumin, analog 2A was detectable in the serum of mice at 30 and 60 minutes after i.p. injections. Analog 2A and curcumin (30 mg/kg bodyweight) showed a similar ability to reduce tumor area in lungs of mice that were i.v. injected with PLS10 cells. Additionally, analog 2A showed superior growth inhibitory effect on PLS10 cells than that of curcumin both in vitro and in vivo. The compound inhibited PLS10 cells growth by induction of G1 phase arrest and apoptosis in vitro. Interestingly, analog 2A significantly decreased tumor growth with downregulation of cell proliferation and angiogenesis in PLS10‐bearing mice. Taken together, we could summarize that analog 2A showed promising activities in inhibiting CRPC progression both in vitro and in vivo.     

姜黄素对复发难治性急性髓细胞白血病患者血清VEGF、survivin表达水平影响

目的:研究姜黄素对复发难治性急性髓细胞白血病患者治疗效果及其可能作用机理.方法:酶联免疫法检测复发难治急性髓细胞白血病患者在不同治疗组中,治疗前后血清VEGF及survivin表达水平.结果:复发难治性急性髓细胞白血病患者血清VEGF、survivin表达水平较对照组显著升高,经过姜黄素胶囊联合化疗治疗后,患者血清VEGF、survivin表达水平显著下降,较单纯化疗组下降更明显,二者差异有统计学意义(P＜0.05).对难治及复发急性白血病患者缓解率影响,二者差异无统计学意义(P＞0.05).结论:姜黄素胶囊联合化疗能显著降低复发难治急性髓细胞白血病患者血清VEGF、survivin表达水平,在抑制急性白血病进展上有一定作用.     

Stereoselective pharmacokinetics of ifosfamide and its 2- and 3-N-dechloroethylated metabolites in female cancer patients

 The pharmacokinetics of the R and S enantiomers of ifosfamide (IFF) and of its 2- and 3-N-dechloroethylated metabolites (2-DCE-IFF and 3-DCE-IFF) were investigated in 14 cancer patients treated with a 3-h infusion of (R,S)-IFF (3 g/m²) with mesna uroprotection. An enantioselective gas chromatographic-mass spectrometric (GC-MS) assay was used to determine the concentrations in plasma and urine. The AUCs of (R)-IFF were significantly larger than those of (S)-IFF (2480±200 vs 1960±150 μM . h). The terminal half-lives (7.57±0.99 h) and mean residence times (11.17±1.10 h) of (R)-IFF were significantly longer than those of (S)-IFF, 6.03±0.82 h and 9.37±0.88 h, respectively. The mean volume of distribution at steady state of (R)-IFF (25.68±0.80 l/m²) was slightly smaller than that of (S)-IFF (27.35±0.89 l/m²). While the renal clearances of (R)-IFF and (S)-IFF were similar, the nonrenal clearance was significantly lower for (R)-IFF (30.20±2.70 vs 41.40±3.55 ml/m² per min) as was total clearance (41.52±2.90 vs 52.37±3.75 ml/m² per min). The AUC values for all of the DCE metabolites from (S)-IFF were significantly greater than those from (R)-IFF with 47% of the measured AUC accounted for by DCE from (S)-IFF compared to only 20% for (R)-IFF. Therefore, the enantioselective difference in IFF elimination can be partially explained by differences in N-dechloroethylation. Received: 13 December 1994/Accepted: 14 May 1995     

Preferential activation of capecitabine in tumor following oral administration to colorectal cancer patients

Purpose: Capecitabine (Xeloda) is a novel fluoropyrimidine carbamate rationally designed to generate 5-fluorouracil (5-FU) preferentially in tumors. The purpose of this study was to demonstrate the preferential activation of capecitabine, after oral administration, in tumor in colorectal cancer patients, by the comparison of 5-FU concentrations in tumor tissues, healthy tissues and plasma. Methods: Nineteen patients requiring surgical resection of primary tumor and/or liver metastases received 1,255 mg/m² of capecitabine twice daily p.o. for 5–7 days prior to surgery. On the day of surgery, samples of tumor tissue, adjacent healthy tissue and blood samples were collected simultaneously from each patient, 2 to 12 h after the last dose of capecitabine had been administered. Concentrations of 5-FU in various tissues and plasma were determined by HPLC. The activities of the enzymes (CD, TP and DPD) involved in the formation and catabolism of 5-FU were measured in tissue homogenates, by catabolic assays. Results: The ratio of 5-FU concentrations in tumor to adjacent healthy tissue (T/H) was used as the primary marker for the preferential activation of capecitabine in tumor. In primary colorectal tumors, the concentration of 5-FU was on average 3.2 times higher than in adjacent healthy tissue (P=0.002). The mean liver metastasis/healthy tissue 5-FU concentration ratio was 1.4 (P=0.49, not statistically different). The mean tissue/plasma 5-FU concentration ratios exceeded 20 for colorectal tumor and ranged from 8 to 10 for other tissues. Conclusions: The results demonstrated the preferential activation of capecitabine to 5-FU in colorectal tumor, after oral administration to patients. This is explained to a great extent by the activity of TP in colorectal tumor tissue, (the enzyme responsible for the conversion of 5′-DFUR to 5-FU), which is approximately four times that in adjacent healthy tissue. In the liver, TP activity is approximately equal in metastatic and healthy tissue, which explains the lack of preferential activation of capecitabine in these tissues. Received: 4 June 1999 / Accepted: 28 October 1999     

Penetration of capecitabine and its metabolites into malignant and healthy tissues of patients with advanced breast cancer

Capecitabine is an oral prodrug of 5-fluorouracil (FU). Since FU concentrations achieved in malignant lesions are an important determinant of efficacy, we investigated the intratumoral transcapillary transfer of capecitabine and its metabolites in vivo. A total of 10 patients with skin metastases from breast cancer received a daily dose of 2500 mg m(-2) capecitabine administered orally in two divided doses for 2 weeks. Microdialysis probes were inserted into a cutaneous metastasis and subcutaneous connective tissue to evaluate the interstitial tissue pharmacokinetics of capecitabine and its metabolites 5'-deoxy-5-fluorocytidine (DFCR), 5'-deoxy-5-fluorouridine (DFUR), and FU by capillary electrophoresis. As intended with the prodrug design of capecitabine, FU was present in low concentrations in tumour interstitium (median c(max): 0.26 microg ml(-1)) when compared with capecitabine, DFCR, and DFUR (median c(max): 2.66, 4.22, and 2.13 microg ml(-1), respectively). Capecitabine and its metabolites easily penetrated malignant and healthy tissue and equilibrated within 45 min between plasma and tissue interstitium. Considering tissue exposure at the extracellular level, no significant differences between healthy and malignant tissues were observed. Our data show that absorption and metabolism determined the tissue pharmacokinetics of capecitabine. There was no evidence of drug tolerance, which may be attributed to impaired transcapillary transfer into tissue, even after repeated administration as shown for three patients.     

姜黄素抑制结肠癌SW480细胞的增殖及其机制

目的:研究姜黄素对人结肠癌SW480细胞增殖以及survivin、caspase-3和p-Akt表达的影响。方法:用不同浓度姜黄素(5～40μmol/L)作用SW480细胞24、48和72 h,MTT法检测姜黄素对SW480细胞生长的抑制作用,Western blotting法检测SW480细胞中survivin、caspase-3和p-Akt蛋白的表达。结果:姜黄素以剂量(5～40μmol/L)和时间(24～72 h)依赖的方式抑制SW480细胞的增殖(P<0.01),40μmol/L姜黄素作用72 h对SW480细胞生长的抑制率可达(75.86±3.93)%。姜黄素抑制SW480细胞中p-Akt、survivin蛋白的表达、促进caspase-3蛋白的表达,均呈时间和剂量依赖性;40μmol/L姜黄素作用SW480细胞48 h后p-Akt和survivin蛋白表达显著减少[(0.204±0.025)vs(0.367±0.035),P<0.01;(0.208±0.014)vs(0.385±0.034),P<0.01],caspase-3蛋白表达显著增加[(0.371±0.028)vs(0.127±0.023),P<0.01]。结论:姜黄素抑制SW480细胞增殖,其机制可能与抑制Akt磷酸化、下调survivin和上调caspase-3蛋白的表达有关。