Arachidonic acid‐containing phosphatidylcholine characterized by consolidated plasma and liver lipidomics as an early onset marker for tamoxifen‐induced hepatic phospholipidosis

Lipid profiling has emerged as an effective approach to not only screen disease and drug toxicity biomarkers but also understand their underlying mechanisms of action. Tamoxifen, a widely used antiestrogenic agent for adjuvant therapy against estrogen‐positive breast cancer, possesses side effects such as hepatic steatosis and phospholipidosis (PLD). In the present study, we administered tamoxifen to Sprague–Dawley rats and used lipidomics to reveal tamoxifen‐induced alteration of the hepatic lipid profile and its association with the plasma lipid profile. Treatment with tamoxifen for 28 days caused hepatic PLD in rats. We compared the plasma and liver lipid profiles in treated vs. untreated rats using a multivariate analysis to determine differences between the two groups. In total, 25 plasma and 45 liver lipids were identified and altered in the tamoxifen‐treated group. Of these lipids, arachidonic acid (AA)‐containing phosphatidylcholines (PCs), such as PC (17:0/20:4) and PC (18:1/20:4), were commonly reduced in both plasma and liver. Conversely, tamoxifen increased other phosphoglycerolipids in the liver, such as phosphatidylethanolamine (18:1/18:1) and phosphatidylinositol (18:0/18:2). We also examined alteration of AA‐containing PCs and some phosphoglycerolipids in the pre‐PLD stage and found that these lipid alterations were initiated before pathological alteration in the liver. In addition, changes in plasma and liver levels of AA‐containing PCs were linearly associated. Moreover, levels of free AA and mRNA levels of AA‐synthesizing enzymes, such as fatty acid desaturase 1 and 2, were decreased by tamoxifen treatment. Therefore, our study demonstrated that AA‐containing PCs might have potential utility as novel and predictive biomarkers for tamoxifen‐induced PLD. Copyright © 2017 John Wiley & Sons, Ltd.     

Discovery of Synthetic Leishmania Inhibitors by Screening of a 2‐Arylbenzothiophene Library

Tamoxifen has been shown to be active in vitro against Leishmania and effective in the treatment for leishmaniasis in murine models. Through the screening of a compound library of estrogen receptor modulator analogs, we identified the major characteristics required for antileishmanial activity. To overcome the difficulties presented by tamoxifen's propensity for E/Z isomerization, we used the 2‐arylbenzothiophene compound BTP as a more stable alternative. Directed screening of a small compound library based on BTP led to active compounds against Leishmania. Subsequent structure–activity data for the synthetic 2‐arylbenzothiophenes evaluated in this study indicate that optimal antileishmanial potency is dependent on the presence of two basic side chains. In addition, the primary structural features required for estrogen receptor binding, the phenols, are not required for inhibiting parasitic growth. Significantly, the most active antileishmanial benzothiophenes lack the pharmacophore for estrogen receptor activity and therefore address potential concerns about the undesirable effects of using selective estrogen receptor modulators in women and children with leishmaniasis. Three compounds selected from the screening have shown consistent activity against all species and stages of Leishmania in vitro although improvements in selectivity are needed. These compounds represent viable starting points for further optimization as antileishmanial agents.     

选择性雌激素受体调节剂的研究进展

雌激素 (ｅｓｔｒｏｇｅｎ)是人体中一类重要的激素类物质。当妇女进入绝经期后 ,体内雌激素水平下降 ,由此引发更年期综合症、骨质疏松、老年性痴呆和心血管疾病等。目前临床上普遍采用替代疗法来治疗或预防这些绝经期易出现的症状。但在治疗中存在的一些问题已经引起医疗界的高度重视 ,如 :阴道出血、乳腺疼痛。尤其是如何确定用替代疗法的最佳时机和需要持续多长时间才能达到最佳治疗效果均尚待解决[1] 。　　以ＩＣＩ 164,384 ( 1)为代表的甾体类雌激素拮抗剂 ,保留了 17β ｅｓｔｒａｄｉｏｌ (ＥＥ2 )的基本骨架 ,表现出纯拮抗作用。 …     

三苯氧胺软膏剂体外透皮释药

目的:测定1%三苯氧胺(TAM)在3种软膏基质中的体外透皮释放,评价三种软膏基质对主药经皮渗透的影响.方法:以人离体皮肤为渗透释药模型,试验用Franz扩散池,λ=235nm处UV测定.结果:在凡士林(Vaseline),聚乙二醇类(PEG)和聚羧乙烯(Carbopol)三种基质中TAM的渗透系数分别在0,1.0677和1.2769μg/(Cm~2h),在24h和46h时的累积释放量分别为0,31.87μg/cm~2和23.48μg/cm~2及0,54.63μg/cm~2和52.44μg/cm~2.结论:TAM在水溶性软膏基质PEG和Carbopol的渗透及释放都优于凡士林软膏基质.     

<Emphasis Type="BoldItalic">In Vitro</Emphasis> Effects on MCF-7 Breast Cancer Cells Of Signal Transduction Inhibitor/Tamoxifen/Eicosapentaenoic Acid Combinations and their Simultaneous Delivery Across Skin

Purpose  To determine the in vitro effects of simultaneously administered LY29400, PD98059, tamoxifen and eicosapentaenoic acid (EPA) on breast cancer cells, and determine their transcutaneous delivery. Methods  Growth assays were performed on MCF-7 cells challenged with IC₅₀ and permeated concentrations of PD98059, LY294002 and tamoxifen firstly in isolation then combined. Permeation studies were performed using PD98059 and LY294002 (singly or simultaneously) in DMSO then fish oil, with enhancers. Immunocytochemical detection of phospho-MAPK, phospho-Akt, total COX-2 and Ki-67 was performed. Results  When applied singly, fluxes of PD98059 and LY294002 were 0.09 ± 0.008 and 0.14 ± 0.045 μg cm⁻² h⁻¹, respectively; applied simultaneously, 0.18 ± 0.045 and 0.49 ± 0.051 μg cm⁻² h⁻¹. Permeated concentrations of PD98059 and LY294002 reduced growth to 13.78 ± 0.63%. Fish oil plus 2.5% DMSO/ethanol allowed 5.96 ± 0.9 and 7.7 ± 1.2 μg cm⁻² of PD98059 and LY294002 to permeate after 48 h. Conclusions  PD98059 and LY294002 permeate excised skin at therapeutically useful rates, and also demonstrate growth inhibitory effects on MCF-7 cancer cells. Synergism was noted in co-transport across skin and activity against cancer cells. A formulation based on fish oil is potentially skin friendly; simultaneous permeation of EPA provides further anti-cancer action.     

他莫昔芬致严重肝损伤

1名44岁女性乳腺癌患者,于手术及化疗后服用他莫昔芬20mg/d作为辅助内分泌治疗。3个月后患者ALT、AST、γ-GT值升高,服用6个月时患者肝功能检查显示:ALT396U/L、AST278U/L、γ-GT211U/L。经保肝治疗后好转,后改用阿那曲唑、戈舍瑞林治疗,未再出现肝功能异常。     

Tamoxifen and risk of idiopathic venous thromboembolism

Aims To evaluate a possible positive association between tamoxifen treatment and the risk of developing idiopathic venous thromboembolism (VTE) in women with breast cancer in the absence of clinical risk factors for venous thromboembolism other than breast cancer itself.
Methods Using information from the large UK-based General Practice Research Database, we identified, within a cohort of more than 10 000 women with breast cancer, all women who developed a first-time diagnosis of deep vein thrombosis or pulmonary embolism of uncertain cause between January 1, 1991 and December 31, 1996. In a case-control analysis, we compared their tamoxifen exposure experience prior to the thromboembolic event with that of a randomly selected group of control women with breast cancer who were matched to cases on age, year of the breast cancer diagnosis and calendar time.
Results We identified 25 cases of idiopathic VTE and 172 controls, all of whom had breast cancer, but were otherwise free from other risk factors for VTE. Past tamoxifen exposure was not materially associated with an elevated risk of developing VTE, and we therefore combined never and past users as reference group. The relative risk estimate of VTE for current tamoxifen exposure, as compared with never and past use combined, was 7.1 (95% CI 1.5–33), adjusted for body mass index, smoking status and hysterectomy status. High body mass index was an independent predictor of VTE itself.
Conclusions Our study provides evidence that current use of tamoxifen increases the risk of idiopathic venous thromboembolism.     

Post‐marketing assessment of generic tamoxifen in Brazilian breast cancer patients

Generic formulations of tamoxifen are commonly prescribed to oestrogen receptor‐positive breast cancer patients at the Brazilian National Cancer Institute (INCA). We carried out a post‐marketing surveillance of the generic tamoxifen formulation in current use at INCA, by comparing plasma concentrations of the parent drug and metabolites obtained with the generic vs the reference formulation. Thirty patients participated in an open‐label, bracketed protocol, comprising 3 successive phases of 30‐32 days each: the generic formulation was used in phases 1 and 3 and the reference formulation in phase 2. Two blood samples were collected in the last 4 days of each phase, for LC‐MS/MS quantification of tamoxifen and metabolites in plasma. The median plasma concentrations (ng/mL) for the reference formulation were as follows: tamoxifen, 135.0 (CI 95% 114.2‐155.8); endoxifen, 35.3 (30.0‐40.8); and 4‐hydroxytamoxifen, 4.8 (4.2‐5.4). The endoxifen/tamoxifen plasma concentration ratio was 0.27 (0.21‐0.25). ANOVA detected no statistically significant difference in plasma concentrations of tamoxifen, metabolites or the endoxifen/tamoxifen ratio among the three phases. The genetic component (rGC) of the CYP2D6‐mediated conversion of tamoxifen into endoxifen, estimated using the repeated drug administration procedure across the three phases, was 0.87, pointing to an important component of genetic variability. In conclusion, this first post‐marketing surveillance trial of oncologic generic drugs carried out in Brazilian patients verified the switchability between the reference and the generic tamoxifen formulation currently used at our institution. The adopted bracketed protocol adds confidence to this conclusion and may serve as a frame for future trials of post‐marketing assessment of other generic drug products.     

三苯氧胺软膏对增生性瘢痕成纤维细胞影响的研究

目的 探讨三苯氧胺抑制增生性瘢痕成纤维细胞的作用.方法 建立兔耳增生性瘢痕动物模型,分别在瘢痕表面外涂三苯氧胺软膏(配制)和空白软膏,观察瘢痕组织、瘢痕增生指数、成纤维细胞数密度变化.结果 (1)实验组比对照组增生性瘢痕体积明显减小、质地较柔软,瘢痕内毛细血管减少,成熟的成纤维细胞较多,排列比较整齐.(2)瘢痕增生指数:对照组(3.32±0.65);实验组(2.73±0.21),2组间差异有显著性(P＜0.05).(3)成纤维细胞数密度:对照组(4 758±306)个/mm2,实验组(3 342±3 111)个/mm2.差异有显著性(P＜0.05).结论 三苯氧胺软膏可抑制增生性瘢痕内成纤维细胞的增殖.     

乳腺癌术后他莫昔芬治疗致脂肪肝

 目的：探讨超声诊断分析乳腺癌术后他莫昔芬治疗致脂肪肝的价值。方法：对217例乳腺癌术后服用他莫昔芬患者进行肝脏彩色多普勒超声检查及胆固醇(CHOL)、甘油三酯(TG)、转氨酶检测。结果：他莫昔芬治疗后,有136例(66%)患者出现脂肪肝。结论：他莫昔芬可以导致肝脏的脂肪变性,发展为脂肪肝,超声检查方便易行,可为临床提供有价值的信息,以便早期诊断、早期治疗。     

三苯氧胺、米非司酮并用米索前列醇抗早孕效果观察

目的　观察三苯氧胺并米索前列醇 (米索 )抗早孕的临床效果 ,同时与米非司酮合并米索作临床随机对比观察 ,评价其可行性 ,为临床提供一种新的抗早孕方法。方法　 1 97例妊娠≤ 56d的妇女 ,按随机表分为两组。结果　研究组完全流产 83例 (84.7% ) ,对照组 90例 (90 .9% ) ,两组比较差异无显著性 ,P >0 .0 5 ;研究组阴道出血时间 (1 0 .3± 5 .1 )d,较对照组 (1 4 .0± 6 .4)d明显缩短 ,1 5d内阴道出血停止者 ,研究组 64例 (77.1 % ) ,对照组 48例 (53.3 % ) ,两组比较差异非常显著 ,P <0 .0 1 :研究组服三苯氧胺后大多数早孕反应未见明显加重 ,而对照组服米非司酮后大多数早孕反应症状进一步加重 ,副反应明显增加 (P <0 .0 1 )。结论　三苯氧胺并米索能有效终止早孕 ,具有抗早孕作用 ,且药物副反应小 ,价格便宜 ,流产后出血时间短 ,完全流产率与米非司酮接近 ,可作为一种抗早孕方法选择使用     

氯离子通道阻断剂对大鼠低氧性肺动脉高压的影响

目的:探讨氯离子通道阻断剂三苯氧胺(tamoxifen,TAM)对大鼠低氧性肺动脉高压的治疗效果。方法:雄性Wistar大鼠96只,随机分为:(1)治疗组(缺氧+三苯氧胺,H/Q):大鼠在常压缺氧(10%,每日8 h)箱内饲养,缺氧前3 d开始灌胃三苯氧胺0.4 mg.kg-1.d-1,每日1次;(2)缺氧组(H组):每日给予相等量的生理盐水,余同治疗组;(3)正常对照组(C组)。测定各组大鼠d 4、7、14、21右心室/(左心室+室间隔)厚度,多导生理记录仪记录肺动脉平均压(mPAP),图像分析仪测定血管管壁面积占管总面积的百分比(WA/TA)和血管管腔面积占管总面积的百分比(VA/TA)。结果:缺氧组d 7mPAP开始升高,d 21达高峰,治疗组同时间点明显低于缺氧组;缺氧组RV/(LV+S)d 14开始上升,d 21达高峰,治疗组同时间点明显低于缺氧组;缺氧组d 21的血管管壁面积占管总面积的百分比(WA/TA)显著高于d 21治疗组(P<0.05)。而缺氧组d 21的血管管腔面积占管总面积的百分比(VA/TA)显著低于d 21治疗组(P<0.05)。结论:氯离子通道阻断剂三苯氧胺可降低缺氧所致大鼠肺动脉压升高、改善右心室肥厚及肺小动脉增厚,对低氧性肺动脉高压有一定的治疗作用。     

他莫昔芬治疗女性绝经后晚期乳腺癌50例

50例晚期乳腺癌妇女,比较国产和英制的他莫昔芬(tamoxifen)的疗效,剂量为10mg bid口服,疗程均在3mo(月)以上,随访6mo以上。结果表明激素受体检测可指导内分泌治疗取得较高效应,2种药物无显著差异。该药对乳腺癌疗效较高,毒副作用较低,安全性大,口服使用方便,优于其他内分泌治疗方案,可为绝经后晚期老年乳腺癌妇女首选的激素治疗药物。     

枸橼酸他莫昔芬口服液的人体药物动力学及相对生物利用度

目的研究枸橼酸他莫昔芬口服液在健康人体内的药物代谢动力学及相对生物利用度。方法以18名健康志愿者为试验对象,采用随机交叉试验方法,分别单剂量口服枸橼酸他莫昔芬口服液2支(含枸橼酸他莫昔芬40 mg)或枸橼酸他莫昔芬片4片(含枸橼酸他莫昔芬40 mg),采用HPLC法测定血浆中药物浓度。结果两种制剂的达峰时间分别为(3.17±0.38)h和(3.94±0.24)h,达峰时血中药物质量浓度分别为(2.1075±0.9685)mg.L-1和(1.8412±0.7723)mg.L-1,两种制剂的消除相半衰期分别为(13.62±1.72)h和(14.09±1.73)h,药时曲线下面积AUC(0→48)分别为(13.4765±2.2966)mg.h.L-1和(14.0920±2.2161)mg.h.L-1,AUC(0→∞)分别为(15.6189±2.5148)mg.h.L-1和(16.1378±2.1349)mg.h.L-1。结论受试制剂与参比制剂两种制剂生物等效。     

枸橼酸他莫昔芬分散片在健康人体的药动学和生物等效性研究

目的：研究枸橼酸他莫昔芬分散片在健康人体内的药动学和生物等效性。方法：采用随机自身对照双周期交叉试验设计，20名男性健康志愿者口服受试制剂和参比制剂各20mg，用高效液相荧光法测定他莫昔芬的血药浓度，用非房室模型计算他莫昔芬的药动学参数。结果：枸橼酸他莫昔芬分散片和普通片的药动学参数如下：tmax分别为（6．3±2．2）、（6．7±2．4）h，Cmax分别为（72±14）、（68±16）μg／L，AUC（0-492）分别为（4．6±2．0）、（4．6±2．0）mg·L^-1·h，t1/2分别为（143±24）、（153±33）h。枸橼酸他莫昔芬分散片相对于普通片的相对生物利用度为（101±13）％。结论：两制剂在人体内生物等效。     

Salinomycin overcomes acquired tamoxifen resistance through AIB1 and inhibits cancer cell invasion in endocrine resistant breast cancer

Salinomycin is a monocarboxylic polyether ionophore isolated from Streptomyces albus. It has been widely used as an antibiotic in veterinary medicine in poultry. A recent study demonstrated that salinomycin selectively inhibits human breast cancer stem cells; one possible mechanism of tamoxifen resistance. Our results show that salinomycin is effective in inhibiting MCF‐7/LCC2 and MCF‐7/LCC9 cell lines which are well‐established endocrine resistant cells and has a synergistic effect in combination with tamoxifen using MTT proliferation assay. The inhibitory effect of salinomycin on the reduction of critical ER co‐activator; amplified breast 1 (AIB1) mRNA and protein expression is overcoming tamoxifen resistance . Moreover, salinomycin significantly inhibits cell invasion in Matrigel invasion assay. The effect was mediated at least in part by the decrease of matrix metalopeptidase 9 (MMP‐9) which is one critical enzyme facilitated in the cell invasion process. In conclusion, salinomycin should be developed as a novel agent used alone or in combination for endocrine‐resistant breast cancer.