Synthesis of trifluoromethyl-containing isoindolinones from tertiary enamides via a cascade radical addition and cyclization process

A radical trifluoromethylation reaction of tertiary enamides was investigated and trifluoromethyl-containing isoindolinones were prepared under mild conditions. Using TMSCF₃ as a radical source, PhI(OAc)₂ as an oxidant and KHF₂ as an additive, tertiary enamides were converted to isoindolinones via a cascade addition and cyclization process in moderate to good yields.

Radical trifluoromethylation and cyclization of tertiary enamides was developed and trifluoromethyl-containing isoindolinones were obtained in moderate to good yields.

In recent years, trifluoromethyl-containing azaheterocycles have attracted much attention for their potential application in the fields of pharmaceutical and agricultural chemistry.¹ Thus, lots of efforts have been devoted to the synthesis of trifluoromethyl azaheterocycles,² and among these developed methods, radical cascade addition and cyclization has emerged as a remarkable strategy due to its unique properties such as economy and high efficiency. Unsaturated amides are commonly used substrates for this type of transformation, which could be attacked by a CF₃ radical followed by intramolecular C–O, C–N, or C–C bond formation to give different kinds of trifluoromethyl azaheterocycles. Fu reported a metal-free trifluoromethylation of N-allyamides with CF₃SO₂Na for the synthesis of trifluoromethyl-containing oxazolines via oxytrifluoromethylation.³ In the presence of copper salts, N-acyl-2-allylaniline could be converted to trifluoromethylated indolines in moderate to good yields via aminotrifluoromethylation process.⁴ With Togni''s reagent,⁵ TMSCF₃,⁶ CF₃SO₂Na,⁷ CF₃SO₂Cl⁸ and other reagents⁹ as the CF₃ source, α, β-unsaturated amides, tosyl amides, or imides underwent a tandem conversion to give trifluoromethyl-containing oxindoles or isoquinoline-1,3-diones by trifluoromethylation/arylation reaction. On the other hand, as a special type of unsaturated amide containing an active double bond, enamide also exhibited excellent reactivity in radical reactions.¹⁰ In fact, trifluoromethylation of enamides has already been investigated, and in most cases trifluoromethylated alkenes were obtained as the main products.¹¹ To the best of our knowledge, the radical trifluoromethylation and cyclization of enamide still remains undeveloped.Isoindolinones are important N-heterocyclic compounds necessary in organic and pharmaceutical chemistry, and these compounds are used widely as anticoagulants and tranquilizers such as aristolactam, pagoclone, and zopiclone.¹² To introduce a CF₃ group into isoindolinones, Wang and co-workers explored a convenient way to the synthesis of trifluoromethyl-containing isoindolinones by radical aminotrifluoromethylation (Scheme 1a),¹³ but this transformation only occurred for N-methoxylbenzamides, and in case of N-alkylbenzamides trifluoromethylated alkenes were obtained as the major products. 1,1-disubstituted terminal alkenes were also not suitable substrates because of the competition between O-trapping and N-trapping process. Thus, development a new method for the synthesis of trifluoromethyl-containing isoindolinones is still in demand. Here in, as a continuation of our efforts on the radical modification of amide derivatives,¹⁴ we wish to present our work on the synthesis of trifluoromethyl-containing isoindolinones using enamides as the start materials by radical carbon trifluoromethylation (Scheme 1b).Open in a separate windowScheme 1Synthesis of trifluoromethyl-containing isoindolinones.Initially, N-n-butyl-N-(2-propenyl) benzamide 1a was chosen as the model substrate to optimize the reaction conditions of this radical carbontrifluoromethylation process. As shown in

Atrial performance in healthy subjects following high altitude exposure at 4100 m: 2D speckle-tracking strain analysis

The International Journal of Cardiovascular Imaging - High altitude (HA) exposure has been considered as a cardiac stress and might impair ventricular diastolic function. Atrial contraction is...     

Antibodies to varicella-zoster virus in blood donors with genetic variance in CC chemokine receptor 5

Carriers of a 32 bp deletion (Delta32) allele of the CC chemokine receptor 5 (CCR5) gene are reported to be more likely to lack antibodies to varicella-zoster virus than CCR5 wild-type individuals. To find out whether CCR5-Delta32 is associated with the seroprevalence of varicella-zoster virus infection, we tested blood donors with different CCR5-Delta32 genotypes for varicella-zoster virus IgG. Antibody to varicella-zoster virus was present in 209 (99.5%) of 210 CCR5-Delta32 carriers and exactly the same proportion of CCR5 wild-type individuals (209 of 210). We have therefore found no evidence that the CCR5-Delta32 allele is associated with decreased seroprevalence of varicella-zoster virus infection.     

诱导痰细胞分类在嗜酸细胞性支气管炎诊断和治疗中的应用

目的探索诱导痰细胞分类在嗜酸细胞性支气管炎(EB)诊断和治疗中的作用。 方法回顾性分析我院门诊239例慢性咳嗽患者,依据慢性咳嗽中诊断标准,分为非EB组和EB组,分析两组诱导痰中细胞分类情况;同时给予EB患者进行8周治疗,观察经治疗后诱导痰中各细胞分类的改变情况。 结果239例慢性咳嗽患者中,非EB患者216例,占90.7%,EB患者23例,占9.3%,两组中共有121例(50.6%)患者诱导痰中嗜酸细胞百分比升高;非EB患者组细胞总计数为(4.83±2.61)×10⁶个/g、嗜酸细胞比例为(4.56±10.07)%、中性粒细胞比例为(50.32±26.12)%、淋巴细胞比例为(5.14±7.27)%、单核巨噬细胞比例为(40.30±16.70)%,EB患者组痰中细胞总计数为(5.46±3.07)×10⁶个/g、嗜酸细胞比例为(13.85±1 2.23)、中性粒细胞比例为(46.16±16.89)、淋巴细胞比例为(4.83±2.98)、单核巨噬细胞比例为(35.91±16.35),两组间EB组嗜酸细胞比例有显著的增多(P<0.05),在性别、年龄以及其余细胞成分差异无统计学意义(P>0.05);EB患者组经治疗后,痰中细胞总计数为(4.07±3.89)×10⁶个/g、嗜酸细胞比例为(2.52±3.80)%、中性粒细胞比例为(54.18±17.97)%、淋巴细胞比例为(5.59±4.33)%、单核巨噬细胞比例为(38.32±17.23)%,嗜酸细胞较治疗前有显著减少(P<0.05),其余细胞改变无显著差异(P>0.05)。 结论诱导痰细胞分类检查是诊断EB的重要方法,在其诊断和治疗过程中有着重要的指导作用,可作为EB治疗过程的监测指标。     

慢性阻塞性肺疾病合并高血压的临床特征及危险因素分析

目的观察慢性阻塞性肺疾病(chronic obstructive pulmonary disease, COPD)合并高血压的临床特征,同时分析其相关的危险因素。 方法收集陆军军医大学新桥医院2016年1月至2018年12月住院的COPD合并高血压患者122例,观察COPD合并高血压的临床特征,同时以单纯COPD患者40例为对照组,应用多因素Logistic回归分析COPD合并高血压的危险因素。 结果122例COPD合并高血压患者中,29.41%合并冠心病、22.99%合并肾疾病和糖尿病、21.39%合并肺心病、14.97%合并脑梗死、4.28%合并心绞痛、4.28%合并心肌梗死、9.63%合并其他心血管疾病。合并冠心病、心肌梗死、其他心血管疾病患者平均年龄显著高于COPD合并高血压患者对照组(P<0.05);合并肺心病患者病程显著长于对照组(P<0.05);合并冠心病、糖尿病、脑梗死、心绞痛、心肌梗死患者每年急性加重次数显著多余对照组(P<0.05);合并心绞痛和其他心血管疾病患者住院时间显著长于对照组(P<0.05)。多因素Logistic回归分析显示年龄(>80岁)、吸烟支数(>700支/年)、饮酒史、CRP(>50 mg/L)、BNP(>100 ng/L)、高血糖是COPD患者合并高血压的危险因素。 结论COPD合并高血压易引起其他相关性疾病而加重病情,及早关注其危险因素可提高患者预后。     

云南省边境地区症状监测预警系统的开发与实践

目的 利用信息化技术,在云南省边境地区建立持续动态症状监测预警系统,并探讨其有效性和及时性,以提升边境地区传染病防控能力。方法 选择3个边境县,以全覆盖的方式,于2016年1月至2018年2月持续监测医疗机构14个症状及6个症候群,收集小学每天学生缺勤信息和边境口岸入境人员发热信息,构建基于手机和电脑平台的症状监测预警系统。结果 采用EARS-3C和Kulldorff时空扫描的预警模型,发现皮疹、流感样症状和小学缺勤等多个监测信号源对手足口病、流感和水痘等常见传染病预警有较高的灵敏度和特异度（可提前1~5 d预警）。系统简便易用,安全性和可行性较强,以交互式图表及可视化地图的方式展示,相关人员能够随时掌握监测数据情况和预警信号的变化,及时采取处置措施。结论 该系统科学有效、操作方便,能实时发现边境地区常见传染病的暴发或聚集事件,实现及时采取有效干预措施,减少本地及跨境传染病暴发的风险,具有实际应用价值。     

mmu_circ_0001033过表达慢病毒载体的构建与鉴定

目的构建环状RNA mmu_circ_0001033慢病毒表达载体,作为在低氧性肺动脉高压功能学实验研究的基础。 方法首先设计引物将目的基因片段从原质粒上扩增下来,通过引物两端所含无缝克隆识别位点将其重组到酶切后的过表达载体上;将连接产物转入制备好的细菌感受态细胞,而后将其单克隆菌落送测序公司进行测序鉴定。其次通过PCR产物跑胶和测序验证成环情况。最后用构建的重组表达载体和包装质粒共同转染293T细胞,包装慢病毒,收集病毒原液,超滤浓缩,并测定滴度。 结果经过比对,重组克隆中插入片段序列与目的片段序列完全一致,表明质粒构建成功。PCR产物后经sanger测序确认环化位点处序列完全正确。将构建的过表达载体测序结果和质粒构建方案基因比对,重组克隆中插入片段序列与目的片段序列与预期完全一致,可实现转染质粒后目的基因的表达,PCR产物跑胶和测序结果表明mmu_circ_0001033成环成功。PCR产物跑胶和测序结果表明mmu_circ_0001033成环成功。最后通过比较RT-qPCR测定病毒滴度值为2.09×10⁸ TU/ml。 结论成功构建环状RNA mmu_circ_0001033慢病毒表达载体,能稳定转染293T细胞,可用于后续细胞实验。     

肠道病毒71型感染患儿心肌损害及心肌酶谱变化分析

目的探讨肌酸激酶同工酶（CK—MB）／肌酸激酶（CK）在肠道病毒71型（EV71）感染病程中对患儿心肌损害的诊断价值。方法选取2010--2014年EV71感染患儿998例,其中CK-MB／CK〉5％的患儿共477例,根据是否存在心肌损害分为心肌损害组（35例）和非心肌损害组（442例）,非心肌损害组按治疗方法分为营养心肌组（209例）和常规治疗组（233例）。在接受常规治疗的基础上,心肌损害组联合营养心肌治疗及其他对症治疗,营养心肌组联合营养心肌治疗。比较各组患儿CK—MB动态变化规律,采用ROC曲线分析CK—MB／CK对心肌损害的预测价值。结果998例EV71感染患儿中约47．8％（477例）伴有CK-MB／CK异常,3．5％（35例）伴有心肌损害。非心肌损害组患儿CK．MB通常在发病的第2-3天达到高峰,持续1～2d,1周左右下降至正常水平;心肌损害组患儿CK—MB高峰则出现在病程的第6～8天,持续4周才缓慢下降至正常水平。营养心肌组和常规治疗组平均治疗时间及病程第7天CK-MB水平比较差异无统计学意义（t=1．767,1．373,P〉0．05）。ROC曲线分析显示CK—MB／CK预测心肌损害曲线下面积为0．638（95％CI：0．554-0．723）,以CK—MB／CK=5．0％为界值,其预测心肌损害的灵敏度为76．5％,特异性为53．2％。结论CK—MB在EV71感染病程中可能存在固有的变化规律,CK-MB／CK对于心肌损害诊断价值不高。     

Apoptotic effect of Aralia echinocaulis extract on fibroblast-like synoviocytes in rats with adjuvant-induced arthritis via inhibiting the Akt/Hif-1α signaling pathway in vitro

Aralia echinocaulis is used for the treatment of rheumatoid arthritis by Tujia Minority in China. A previous study demonstrated that A. echinocaulis had a significant anti-arthritic effect on adjuvant arthritis (AA) rats in vivo. However, it remains unclear whether A. echinocaulis can induce the apoptosis of fibroblast-like synoviocytes (FLS) from AA rats and the underlying mechanism is unknown. In this paper, CCK-8 assay, Hoechst staining and flow cytometry were used to evaluate the apoptotic effect of an A. echinocaulis ethanol extract (AEE) on AA FLS. Western blotting analysis was performed to measure the protein expression levels of Bcl-2, Bax, cleaved caspase-3, Akt, p-Akt, and Hif-1α. The results revealed that AEE could inhibit FLS proliferation in a dose and time-dependent manner. After treatment with AEE, AA FLS displayed the classical apoptotic morphology, and the apoptosis rates were significantly increased. Furthermore, we found that AEE increased the protein levels of Bax, cleaved caspase 3, and decreased the protein levels of Bcl-2, Hif-1α and p-Akt, without affecting total Akt levels. Collectively, these results suggested that the apoptosis inducing effect of AEE on AA FLS was related to the regulation of the expression of apoptosis-related proteins and the inhibition of the Akt/Hif-1α signaling pathway.     

基因芯片筛选Sirt1在小鼠急性呼吸窘迫综合征炎症损伤中相互作用的基因

目的筛选小鼠急性呼吸窘迫综合征炎症损伤中与Sirt1相互作用的基因,探讨Sirt1在急性呼吸窘迫综合征炎症反应中的作用机制。方法购买Sirt1过表达(Tg)小鼠和野生型(WT)小鼠,饲养、繁殖、鉴定新生小鼠基因型;Western Blot鉴定小鼠肺组织Sirt1表达差异;建立ARDS小鼠模型,观察ARDS小鼠肺组织HE染色病理变化,并采用ELISA检测两种ARDS小鼠肺组织IL-6表达差异;采用基因芯片筛选Sirt1在小鼠ARDS炎症损伤中相互作用的基因。结果通过聚合酶链反应鉴定出Tg和WT两种不同基因型的小鼠;免疫印迹法检测小鼠肺组织Sirt1的表达差异结果提示Tg小鼠肺组织Sirt1含量显著高于WT小鼠(P=0.001);不同浓度LPS腹腔注射12 h后小鼠肺组织HE染色病理变化提示随着LPS用量增加,两种小鼠肺组织损伤程度明显增加,且WT-ARDS小鼠的肺组织损伤程度比Tg-ARDS小鼠更为严重;当LPS用量达到20 mg/kg时两组小鼠存活率50%;两种小鼠腹腔注射LPS(15 mg/kg)后,肺组织IL-6的表达随时间推移逐渐呈现逐渐增高的趋势,在3,6,12,24 h WT-ARDS组肺组织IL-6表达浓度显著高于Tg-ARDS组(P0.05),尤其在12 h差异最为显著(P=0.007)。基因芯片筛选出在小鼠ARDS炎症损伤中与Sirt1相互作用的基因有Ubd,Ube2d2b,Olfm4,Il1rl1,Hivep3和Lpar1。结论 Sirt1可能通过调控Ubd,Ube2d2b,Olfm4,Il1rl1,Hivep3和Lpar1的表达减轻ARDS小鼠的炎症损伤。