Design,synthesis,and evaluation of fluoroquinolone derivatives as microRNA-21 small-molecule inhibitors

MicroRNA-21(miRNA-21)is highly expressed in various tumors.Small-molecule inhibition of miRNA-21 is considered to be an attractive novel cancer therapeutic strategy.In this study,fluoroquinolone derivatives A1eA43 were synthesized and used as miRNA-21 inhibitors.Compound A36 showed the most potent inhibitory activity and specificity for miRNA-21 in a dual-luciferase reporter assay in HeLa cells.Compound A36 significantly reduced the expression of mature miRNA-21 and increased the protein expression of miRNA-21 target genes,including programmed cell death protein 4(PDCD4)and phosphatase and tensin homology deleted on chromosome ten(PTEN),at 10 μM in HeLa cells.The Cell Counting Kit-8 assay(CCK-8)was used to evaluate the antiproliferative activity of A36;the results showed that the IC_(50) value range of A36 against six tumor cell lines was between 1.76 and 13.0 μM.Meanwhile,A36 did not display cytotoxicity in BEAS-2B cells(lung epithelial cells from a healthy human donor).Furthermore,A36 significantly induced apoptosis,arrested cells at the G_(0)/G_(1) phase,and inhibited cell-colony formation in HeLa cells.In addition,mRNA deep sequencing showed that treatment with A36 could generate 171 dysregulated mRNAs in HeLa cells,while the expression of miRNA-21 target gene dual-specificity phosphatase 5(DUSP5)was significantly upregulated at both the mRNA and protein levels.Collectively,these findings demonstrated that A36 is a novel miRNA-21 inhibitor.

Bioactivities,Mechanisms,Production,and Potential Application of Bile Acids in Preventing and Treating Infectious Diseases

Infectious diseases are a global public health problem,with emerging and re-emerging infectious diseases on the rise worldwide.Therefore,their prevention and treatment are still major challenges.Bile acids are common metabolites in both hosts and microorganisms that play a significant role in controlling the metabolism of lipids,glucose,and energy.Bile acids have historically been utilized as first-line,valuable therapeutic agents for related metabolic and hepatobiliary diseases.Notably,bile acids are the major active ingredients of cow bezoar and bear bile,which are commonly used traditional Chinese medicines(TCMs)with the therapeutic effects of clearing heat,detoxification,and relieving wind and spasm.In recent years,the promising performance of bile acids against infectious diseases has attracted attention from the scientific community.This paper reviews for the first time the biological activities,possible mechanisms,production routes,and potential applications of bile acids in the treatment and prevention of infectious diseases.Compared with previous reviews,we comprehensively summarize existing studies on the use of bile acids against infectious diseases caused by pathogenic microorganisms that are leading causes of global morbidity and mortality.In addition,to ensure a stable supply of bile acids at affordable prices,it is necessary to clarify the biosynthesis of bile acids in vivo,which will assist scientists in elucidating the accumulation of bile acids and discovering how to engineer various bile acids by means of chemosynthesis,biosynthesis,and chemoenzymatic synthesis.Finally,we explore the current challenges in the field and recommend a development strategy for bile-acid-based drugs and the sustainable production of bile acids.The presented studies suggest that bile acids are potential novel therapeutic agents for managing infectious diseases and can be artificially synthesized in a sustainable way.

Study of isolation of fluoroquinolone-resistant Ureaplasma urealyticum and identification of mutant sites

OBJECTIVE: To study the resistance mechanism of clinical isolates of Ureaplasma urealyticum resistant to fluoroquinolones. METHODS: Thirteen isolates of Ureaplasma urealyticum resistant to six fluoroquinolones were selected out of 184 clinical isolates and their QRDRs (quinolone resistance-determining region) gyrA, gyrB, parC and parE were amplified by PCR. Sequencing results were compared to those susceptible reference strains and a comparison of deduced amino acid sequences were performed. RESULTS: Sequence comparison revealed a C to A change at 87nt of gyrA QRDR leading to the substitution of Asp95 with glutamic acid and a C to T change at 50nt of parC QRDR leading to the substitution of Ser80 with leucine. CONCLUSION: These results suggest that a C to A change at 87nt of gyrA QRDR and a C to T change at 50nt of parC QRDR are associated with fluoroquinolone resistance of Ureaplasma urealyticum.

氧氟沙星的荧光光谱与质子化作用研究

研究了氧氟沙星(Ofloxacin ,OFL)在不同pH条件下的荧光光谱、紫外光谱和质子化作用。在强酸性溶液中,OFL分子可以结合两个质子而以三元酸H3 L2 + 的形式存在,最大荧光发射波长(λmax)为5 0 5nm。随着pH值升高,OFL的荧光光谱发生变化,在激发光谱中35 2nm形成一等荧光点,同时在紫外吸收光谱中出现等色点,这一光谱特征表明H3 L2 + 逐渐失去4位C羰基氧结合的质子。在pH 2 . 5～4范围内,OFL以H2 L+ 形式存在,λmax为4 99nm。当pH >4时,随pH值升高,位于4 99nm的荧光发射峰逐渐蓝移至4 5 5nm ,在4 84nm形成一等荧光发射点,表明C 3位羧基质子的离解。在pH 7左右,OFL以双极离子HL形式存在,λmax为4 5 5nm ,是最强的荧光型体。当pH >8时,随pH值升高,λmax由4 5 5nm红移至约4 75nm ,同时荧光强度下降,表明HL失去哌嗪环N -4上结合的质子。当pH >10时,OFL以阴离子L-形式存在,荧光强度随pH值升高而降低,但λmax基本不变,表明介质环境对OFL的荧光性质有一定影响。

我国喹诺酮类抗感染药物的研究进展

对我国近几年喹诺酮类药物研究开发概况、新合成路线与工艺改进、结构改造与构效关系研究进行了简要的述评。

近年我国喹诺酮类抗菌剂的研究进展

对近几年我国喹诺酮类抗菌剂研究开发情况,包括合成工艺改进以及创新药物的研究进行了简要综述。

120株临床分离鲍曼不动杆菌对喹诺酮类耐药表型及其机制

目的了解鲍曼不动杆菌对喹诺酮类抗菌药物的常见耐药性及其机制,为临床抗感染治疗提供依据,以指导临床合理用药。方法采用VITEK系统鉴定细菌,用M-H琼脂定量稀释法测定菌株对左氧氟沙星、环丙沙星和加替沙星3种喹诺酮类药物的最低抑菌(MIC),PCR扩增gyrA和parC基因,选取部分PCR扩增产物进行测序分析。结果临床标本共检出120株鲍曼不动杆菌,中心重症监护室、呼吸科病房、急救中心分离菌所占比率分别为28.3%、24.2%、12.5%;对环丙沙星、左氧氟沙星和加替沙星的耐药率分别为78.3%、58.3%和40.0%。PCR基因扩增和测序结果:所有菌株均扩增出了相应gyrA和parC基因产物;耐药株中的gyrA基因存在83位氨基酸密码子突变,敏感株中则不存在该突变,并在耐药株中发现gyrA和parC基因针对GenBank号分别为DQ270238和EU977576的菌株存在新的点突变。结论鲍曼不动杆菌对喹诺酮类药物耐药情况严重,其耐药性与gyrA和parC基因点突变有关。

加替沙星注射液的研制和质量控制

目的 :探讨加替沙星注射液的处方、制备工艺及质量控制方法。方法 :以盐酸、氢氧化钠为 pH调节剂及氯化钠为等渗调节剂制备加替沙星注射液 ;采用高效液相色谱法测定本品的含量。结果 :制备的加替沙星注射液 (pH 5 .0 )在高温 (80°C)、光照(3 60 0lx) 1 0d及室温留样考察 2 4mo ,物理及化学性质稳定。结论 :处方及工艺经中试放大制备的注射液性质稳定 ;质量控制方法准确可靠 ;可满足临床用药要求。

南京地区肺炎链球菌的耐药变迁及喹诺酮耐药机制研究

目的了解南京地区肺炎链球菌临床分离株的耐药性变迁及对喹诺酮类的耐药机制。方法收集2010—2012年南京7所教学医院共147株肺炎链球菌,琼脂稀释法测定9种抗菌药物的MIC,与南京2006—2007年分离的肺炎链球菌耐药情况进行比较。并对氟喹诺酮耐药株的gyrA、gyrB、parE和parC基因喹喏酮耐药决定区域(QRDR)进行PCR扩增及测序。结果 147株肺炎链球菌中,对红霉素的耐药率为89.1%;青霉素不敏感肺炎链球菌(MIC≥4 mg/L)占3.4%;对头孢呋辛、头孢曲松、美罗培南、左氧氟沙星、莫西沙星的耐药率分别为26.5%、1.4%、3.4%、1.4%、0.7%;所有分离株对万古霉素、利奈唑胺敏感。与2006—2007年分离株相比,2010—2012年临床分离株对青霉素、头孢呋辛耐药率下降,对红霉素、头孢曲松耐药率变化不大,出现了少数左氧氟沙星及莫西沙星耐药株。对喹诺酮耐药株进行基因测序发现1菌株有gyrA突变(Asn167—Ile)和ParE突变(Ile 460—Val);另1菌株则有ParC突变(Ser 81—Gly;Asn 94—Asp)。结论南京地区肺炎链球菌对红霉素耐药率居高不下,对青霉素仍较敏感,出现了少数莫西沙星、左氧氟沙星耐药株。喹诺酮耐药株有gyrA、parE和parC的QRDR突变。

临床分离鲍曼不动杆菌耐药表型和质粒介导喹诺酮类耐药基因研究

目的:对医院分离出的鲍曼不动杆菌耐药谱进行调查,探求鲍曼不动杆菌中质粒介导对喹诺酮类药物耐药基因(PMQR)携带情况,给临床感染性疾病治疗提供参考。方法:采用VITEKT-2 compact全自动细菌鉴定及药敏系统对菌株进行鉴定和药敏检测,采用PCR法检测细菌是否携带PMQR基因(qnrA、B、C、D、S、aac(6')-Ib和qepA),随机选取每种耐药基因的PCR产物进行测序分析。病原菌的药敏结果采用WHONET 5.6软件分析。结果:近两年来从临床标本中一共检出152株鲍曼不动杆菌,药敏结果显示鲍曼不动杆菌对喹诺酮类和其他抗菌药物的耐药性有逐年递增之势。PCR检测结果显示29.6%(45/152)菌株携带aac(6')-Ib,1.3%(2/152)菌株携带qnrB,质粒上的qnrA、C、D、S和外排泵qepA基因产物并未检测出。结论:本地区鲍曼不动杆菌对临床常用抗菌药物的耐药性强,PMQR携带率较低。

喹诺酮类药物与碳青霉烯类药物之间的交叉耐药性和泛耐药性的研究

目的研究由喹诺酮类和碳青霉烯类药物诱导的耐药性铜绿假单胞菌之间的交叉耐药,以及与庆大霉素(CN)、头孢哌酮/舒巴坦(SCF)之间的交叉耐药情况。方法用左氧氟沙星(LEV)和亚胺培南(IPM)分别诱导3株铜绿假单胞菌菌株(1株为标准菌株,2株为野生菌株),使之耐药,通过检测LEV、IPM、CN、SCF对该菌株的最低抑菌浓度(MIC),与未耐药菌株的MIC进行对比,观察药物间的交叉耐药情况。并通过对2005—2008年的铜绿假单胞菌耐药情况的分析,进一步观察上述药物间的交叉耐药情况。结果LEV、IPM、CN、SCF对由LEV诱导耐药的铜绿假单胞菌标准菌株的MIC值分别为32、16、16、32μg/ml,野生菌株的MIC值分别为16、8、8、64μg/ml和8、16、16、32μg/ml;LEV、IPM、CN、SCF对由LEV诱导耐药的铜绿假单胞菌标准菌株的MIC值分别为16、16、128、64μg/ml,野生菌株的MIC值分别为8、8、8、64μg/ml;和4、16、8、64μg/ml。结论由LEV、IPM诱导耐药的铜绿假单胞菌菌株,对LEV、IPM、CN、SCF均产生了不同程度的耐药性。提示喹诺酮类与碳青霉烯类药物的滥用都可能导致广泛的细菌交叉耐药。

蒙脱石散对部分喹诺酮类药物的体外吸附作用

目的 :比较在人工胃液和水溶液中蒙脱石散对常用剂量的喹诺酮类药物吡哌酸、诺氟沙星、氧氟沙星、盐酸环丙沙星、甲磺酸培氟沙星、盐酸洛美沙星的吸附作用。方法 :取蒙脱石散分别加入到上述药物的水溶液和人工胃液中 ,用紫外分光光度法测定药物含量 ,计算吸附率。结果 :在水溶液中吡哌酸、诺氟沙星、氧氟沙星极微溶。在人工胃液中 ,药物质子化程度提高 ,表现为阳离子特性 ,蒙脱石对其吸附率达 98.6 %以上。盐酸环丙沙星、甲磺酸培氟沙星、盐酸洛美沙星已与酸成盐 ,无论在水溶液还是在人工胃液中 ,蒙脱石对其的吸附率达 99%以上。结论 :蒙脱石对常用剂量的喹诺酮类药物有强大的吸附作用 ,服用上述药物应在服用蒙脱石散之后 1～ 2h。

我国喹诺酮类抗感染药物的研究开发进展

对我国喹诺酮类药物已开发产品的概况、新合成路线与工艺改进、结构改造与构效关系研究以及环丙氟啶酸的临床前研究作了较系统的述评。

新抗菌药研究开发的进展

β-内酰胺类抗生素特别是头孢菌素在抗生素领域里仍占主导地位，碳青霉烯类的发展速度比以前更快。本综述包括11个部分：青霉素类、头孢菌素类、碳青霉烯类、青霉烯类、其他β-内酰胺类、氨基糖苷类、大环内酯类、四环素类、安莎霉素类、喹诺酮类与其他合成抗菌药。

抗菌药研究的进展(一)

综述了近年来抗菌药研究的进展。包括１３个部分：头孢菌素类，碳青霉烯类，青霉烯类，β内酰胺酶抑制剂，β内酰胺增强剂，具有其他药理活性的β内酰胺，氨基糖苷类，大环内酯类，四环素类，安莎类抗生素，糖肽类与链阳菌素类，喹诺酮类，其他合成抗菌药。

莫西沙星治疗成人支原体肺炎的初步探讨

【目的】探讨莫西沙星对支原体肺炎的治疗作用。【方法】回顾性分析20例经莫西沙星治疗的支原体肺炎患者,通过患者体温变化、CT影像变化探讨莫西沙星对支原体肺炎的疗效。【结果】①患者入院时的平均体温为:(38.50±0.86)℃,治疗后d3和d5时,体温分别为:(37.18±0.61)℃和(36.77±0.35)℃,分别下降了(1.32±0.81)℃和(1.69±0.76)℃,与入院时比较均有显著的统计学差异(P均<0.001)。体温在治疗d3和d5之间,亦存在显著性差异(P<0.01)。②患者的平均住院时间为(13.56±3.96)d。③本组痊愈5例(5/13);显效6例(6/13);进步2例(2/13);无效0例(0/13)。【结论】莫西沙星静脉滴注治疗成人支原体肺炎,在控制体温和炎症吸收上可能具有较好的疗效。

Low eradication rate of Helicobacterpyloriwith triple 7-14 days and quadriple therapy in Turkey

AIM:The eradication rate of Helicobacter pylori (H pylori) shows variation among countries and regimens of treatment. We aimed to study the eradication rates of different regimens in our region and some factors affecting the rate of eradication. METHODS:One hundred and sixty-four H pylori positive patients (68 males,96 females;mean age:48±12 years) with duodenal or gastric ulcer without a smoking history were included in the study.The patients were divided into three groups according to the treatment regimens.Omeprazole 20mg,clarithromycin 500mg,amoxicillin 1g were given twice daily for 1 week (Group Ⅰ) and 2 weeks (Group Ⅱ). Patients in Group Ⅲ received bismuth subsitrate 300mg, tetracyline 500mg and metronidazole 500mg four times daily in addition to Omeprazole 20mg twice daily.Two biopsies each before and after treatment were obtained from antrum and corpus,and histopathologically evaluated. Eradication was assumed to be successful if no H pylorus was detected from four biopsy specimens taken after treatment.The effects of factors like age,sex,H pylori density on antrum and corpus before treatment,the total H pylori density,and the inflammation scores on the rate of H pylori eradication were evaluated. RESULTS:The overall eradication rate was 42%.The rates in groups Ⅱ and Ⅲ were statistically higher than that in group Ⅰ (P<0.05).The rates of eradication were 24.5%, 40.7% and 61.5% in groups Ⅰ,Ⅱ and Ⅲ,respectively.The eradication rate was negatively related to either corpus H pylori density or total H pylori density (P<0.05).The median age was older in the group in which the eradication failed in comparison to that with successful eradication (55 yr vs 39 yr,P<0.001).No correlation between sex and H pylori eradication was found. CONCLUSION:Our rates of eradication were significantly lower when compared to those reported in literature.We believe that advanced age and high H pylori density are negative predictive factors for the rate of H pylori eradication.

对贵刊《加替沙星与左氧氟沙星治疗泌尿道感染的疗效比较》的验证

笔者阅读了贵刊2004年第23卷第9期611页刊登的《加替沙星与左氧氟沙星治疗泌尿道感染的疗效比较》一文后，深受启发，并采用此法，于2004年10月～2005年9月对我校医院门诊60例泌尿道感染病人进行了临床验证，结果报道如下。临床资料从本校医院门诊泌尿道感染病人选择60例作为观察对象，分为2组。2组病人年龄、性别、病程、感染严重程度等方面经X^2或t检验，差异无显著意义（P〉0．05）。

抗菌新药甲磺酸加替沙星的药效学实验研究

目的 :研究抗菌新药甲磺酸加替沙星粉剂的体外抗菌效果和体内保护作用 ,并与国家药品监督管理局已批准进入临床研究的加替沙星粉剂进行比较。方法 :按照我国二类新药抗生素类药效学实验指导原则 ,选用试管对倍稀释法作体外抗菌实验及KM小白鼠作体内保护实验。结果 :甲磺酸加替沙星和阳性对照药加替沙星对 14种 (2 3 2株 )临床菌株和质控菌株的最小抑菌浓度 (MIC)值、最小杀菌浓度 (MBC)值及杀菌速率均无显著差异。不同细菌接种量和不同培养基pH值对两药MIC值的影响也基本一致。体内保护实验显示 ,两药对金黄色葡萄球菌、铜绿假单胞菌感染的小鼠模型均具有很强的保护作用 ,且两者的ED50 及 95 %可信限基本相同。结论

高效液相法测定加替沙星尿药浓度

目的 建立反相高效液相色谱法以测定大鼠尿液中加替沙星浓度。方法 尿液样品用二氯甲烷提取 ,以环丙沙星为内标 ,色谱柱 :LichrosorbC1 8,流动相 :甲醇 - 0 .0 1mol/L ,磷酸二氢钾缓冲液 - 0 .5mol/L ,四丁基氢氧化铵溶液 (1 5∶75∶1 .4) ,磷酸调 pH至 2 .6 ;流速为 1 .0ml/min ;检测波长 2 98nm。 结果 本法在 3 .1 0～ 1 55 .2 μg/ml范围内线性良好 ,r=0 .9995 ,日内RSD为 5 .1 %～ 9.7% ,日间RSD为 5 .7%～ 1 0 .2 % (n =5) ;尿液最低检测浓度为 50ng/ml。 结论 本法测试快速、准确。