Discovery of a new HIV-1 inhibitor scaffold and synthesis of potential prodrugs of indazoles

A new oxazole scaffold showing great promise in HIV-1 inhibition has been discovered by cell-based screening of an in-house library and scaffold modification. Follow-up SAR study focusing on the 5-aryl substituent of the oxazole core has identified 4k (EC₅₀ = 0.42 μM, TI = 50) as a potent inhibitor. However, the analogues suffered from poor aqueous solubility. To address this issue, we have developed broadly applicable potential prodrugs of indazoles. Among them, N-acyloxymethyl analogue 11b displayed promising results (i.e., increased aqueous solubility and susceptibility to enzymatic hydrolysis). Further studies are warranted to fully evaluate the analogues as the potential prodrugs with improved physiochemical and PK properties     

A convenient synthesis of lubeluzole and its enantiomer: Evaluation as chemosensitizing agents on human ovarian adenocarcinoma and lung carcinoma cells

Lubeluzole, a neuroprotective anti-ischemic drug, and its enantiomer were prepared following a convenient procedure based on hydrolytic kinetic resolution. The ee values were >99% and 96%, respectively, as assessed by HPLC analysis. The chemosensitizing effects of both enantiomers were evaluated in combination with either doxorubicin (human ovarian adenocarcinoma A2780 cells) or paclitaxel (human lung carcinoma A549 cells) by the MTT assay. At the lowest concentrations used, lubeluzole showed an overall and remarkable tendency to synergize with both anticancer drugs. In ovarian cancer cells a clear prevalence of antagonistic effect was observed for the R-enantiomer. The synergistic effects of lubeluzole for both drugs were observed over a wide concentration window (0.005–5 μM), the lowest limit being at least 40 times lower than human plasma concentrations previously reported as causing serious side effects.     

3,5-Bis(benzylidene)-4-piperidones and related N-acyl analogs: A novel cluster of antimalarials targeting the liver stage of Plasmodium falciparum

Drug resistance is a major challenge in antimalarial chemotherapy. In addition, a complete cure of malaria requires intervention at various stages in the development of the parasite within the host. There are only a few antimalarials that target the liver stage of the Plasmodium species which is an essential part of the life cycle of the malarial parasite. We report a series of antimalarial 3,5-bis(benzylidene)-4-piperidones and related N-acyl analogs 1–5, a number of which exhibit potent in vitro growth-inhibiting properties towards drug-sensitive D6 and drug-resistant C235 strains of Plasmodium falciparum as well as inhibiting the liver stage development of the malarial life cycle. The compounds 2b (IC₅₀: 165 ng/mL), 3b (IC₅₀: 186 ng/mL), 5c (IC₅₀: 159 ng/mL) and 5d (IC₅₀: 93.5 ng/mL) emerged as lead molecules that inhibit liver stage Plasmodium berghei and are significantly more potent than chloroquine (IC₅₀: >2000 ng/mL) and mefloquine (IC₅₀: >2000 ng/mL) in this screen. All the compounds that showed potent inhibitory activity against the P. berghei liver stage were nontoxic to human HepG2 liver cells (IC₅₀: >2000 ng/mL). The compounds 5a and 5b exhibit comparable metabolic stability as chloroquine and mefloquine in human plasma and the most potent compound 5d demonstrated suitable permeability characteristics using the MDCK monolayer. These results emphasize the value of 3,5-bis(benzylidene)-4-piperidones as novel antimalarials for further drug development.     

Defining RNA motif–aminoglycoside interactions via two-dimensional combinatorial screening and structure–activity relationships through sequencing

RNA is an extremely important target for the development of chemical probes of function or small molecule therapeutics. Aminoglycosides are the most well studied class of small molecules to target RNA. However, the RNA motifs outside of the bacterial rRNA A-site that are likely to be bound by these compounds in biological systems is largely unknown. If such information were known, it could allow for aminoglycosides to be exploited to target other RNAs and, in addition, could provide invaluable insights into potential bystander targets of these clinically used drugs. We utilized two-dimensional combinatorial screening (2DCS), a library-versus-library screening approach, to select the motifs displayed in a 3 × 3 nucleotide internal loop library and in a 6-nucleotide hairpin library that bind with high affinity and selectivity to six aminoglycoside derivatives. The selected RNA motifs were then analyzed using structure–activity relationships through sequencing (StARTS), a statistical approach that defines the privileged RNA motif space that binds a small molecule. StARTS allowed for the facile annotation of the selected RNA motif–aminoglycoside interactions in terms of affinity and selectivity. The interactions selected by 2DCS generally have nanomolar affinities, which is higher affinity than the binding of aminoglycosides to a mimic of their therapeutic target, the bacterial rRNA A-site.     

Cynaropicrin targets the trypanothione redox system in Trypanosoma brucei

In mice cynaropicrin (CYN) potently inhibits the proliferation of Trypanosoma brucei—the causative agent of Human African Trypanosomiasis—by a so far unknown mechanism. We hypothesized that CYNs α,β-unsaturated methylene moieties act as Michael acceptors for glutathione (GSH) and trypanothione (T(SH)₂), the main low molecular mass thiols essential for unique redox metabolism of these parasites. The analysis of this putative mechanism and the effects of CYN on enzymes of the T(SH)₂ redox metabolism including trypanothione reductase, trypanothione synthetase, glutathione-S-transferase, and ornithine decarboxylase are shown. A two step extraction protocol with subsequent UPLC–MS/MS analysis was established to quantify intra-cellular CYN, T(SH)₂, GSH, as well as GS-CYN and T(S-CYN)₂ adducts in intact T. b. rhodesiense cells. Within minutes of exposure to CYN, the cellular GSH and T(SH)₂ pools were entirely depleted, and the parasites entered an apoptotic stage and died. CYN also showed inhibition of the ornithine decarboxylase similar to the positive control eflornithine. Significant interactions with the other enzymes involved in the T(SH)₂ redox metabolism were not observed. Alongside many other biological activities sesquiterpene lactones including CYN have shown antitrypanosomal effects, which have been postulated to be linked to formation of Michael adducts with cellular nucleophiles. Here the interaction of CYN with biological thiols in a cellular system in general, and with trypanosomal T(SH)₂ redox metabolism in particular, thus offering a molecular explanation for the antitrypanosomal activity is demonstrated. At the same time, the study provides a novel extraction and analysis protocol for components of the trypanosomal thiol metabolism.     

尿液中奇异变形杆菌耐药性变迁及监测分析

目的了解尿液中奇异变形杆菌临床分布及耐药性变迁情况,为临床医生合理选用抗生素提供理论依据。方法收集2008年至2012年住院及门诊患者尿液标本中分离的215株奇异变形杆菌,采用VITEK2 Compact全自动微生物分析仪进行鉴定及药敏试验,采用WH0NET 5.4软件进行统计分析。结果2008、2009、2010、2011和2012年分离奇异变形杆菌分别为26株（占12. 2% ）、26株（占12.2% ）、36株（占16. 9% ）、55株（占25. 8% ）和72株（占33. 8% ）,总计215株。奇异变形杆菌对呋喃妥因的耐药率最高,均〉90%,对丁胺卡那霉素和美洛培南的耐药率最低,均为0%,对左旋氧氟沙星的耐药率在2008？2011年均〉50% ,2012年为28.1% ;对哌拉西林/他唑巴坦的耐药率在2008 - 2009年均〉20%,而2010-2012年均〈5% ;头孢替坦、舒普深耐药率均〈7%。结论奇异变形杆菌的分离率从2008 -2012年呈日益增长趋势,提醒我们随着临床上大量使用抗生素,医院的感染率亦不断上升。耐药率从2008-2011年基本呈上升趋势,而在2012年有所下降,这可能与近几年本院临床严格执行抗菌药物的合理应用有关。     

泌尿系感染常见病原菌的分布及耐药性分析

【摘 要】 目的 探讨解放军第44医院住院患者泌尿系感染病原菌的分布及耐药性,为临床合理应用抗菌药物提供参考。方法 通过法国生物梅里埃VITEK 2 compact及ATB-Expression细菌鉴定分析仪对2012年该院收治的698例住院患者送检中段尿标本进行细菌鉴定,并用K-B法对分离病原菌进行体外药敏试验。结果 698例患者中有512例培养阳性,其中女性患者阳性率为81.5%,男性患者阳性率为19.0%,二者比较差异有统计学意义(P＜0.05)。512例阳性患者中段尿标本中共分离出病原菌536株,其中革兰阴性杆菌占68.8%,革兰阳性球菌占22.8%,真菌占8.4%。分离病原菌对常用抗菌药物呈现多重耐药性,革兰阴性杆菌对氨苄西林、哌拉西林、头孢噻吩、复方新诺明的耐药率高于60.0%,对亚胺培南、阿米卡星敏感。革兰阳性球菌对克林霉素、红霉素、青霉素的耐药率高于70.0%,对万古霉素、替考拉宁高度敏感。结论 泌尿系感染病原菌对常用抗菌药物耐药严重,及时了解泌尿系感染病原菌的分布特点及耐药性,对临床合理选用抗菌药物具有重要意义。     

医院感染病原菌分布及药物敏感性分析

【摘 要】 目的 探讨医院感染病原菌的分布及其药物敏感性,以指导临床合理用药。方法 对金华市中心医院2009年至2011年临床分离的1 693株病原菌及药物敏感性进行回顾性分析。结果 医院感染病原菌主要是G-菌,占60.7%,G+菌占23.8%;感染部位以呼吸道、泌尿道为主,易感人群主要是血液系统疾病、肝病肝硬化患者,科室主要分布于重症监护病房、放疗科、血液科。G+菌对常用抗菌药耐药严重,但对利福平、呋喃妥因、万古霉素敏感率相对高;常见G-菌对氨基糖甙类、碳青酶烯类、β-内酰胺酶抑制剂敏感率较高。结论 医院感染监控应从感染科室、感染部位、易感人群等多方面进行,及时动态了解医院感染病原菌分布及其药物敏感性,利于指导合理用药。     

儿童呼吸道感染时呼吸道分泌物的病原菌分布及其耐药性分析

【摘 要】目的 了解本地区儿童呼吸道感染病原菌种类、分布特点及其耐药性,为临床合理应用抗菌药物提供参考。方法 对2009年5月至2012年5月间本地区两家医院儿科住院患者送检的共1350份呼吸道分泌物的细菌培养结果及要检出菌的耐药情况,进行回顾性分析。结果 从1350份标本中共分离出384株病原菌,检出率为28.4%,革兰阴性杆菌258株,占67.2%,革兰阳性球菌95株,占24.7%,厌氧菌17株,占4.4%,真菌14株,占3.6%;革兰阴性前三位杆菌肺炎克雷伯杆菌、大肠埃希菌和解鸟氨酸克雷白杆菌对氨苄西林、哌拉西林耐药率均大于50%以上,共分离出产ESBLs菌57株,阳性率为43.2%;革兰阳性前三位球菌肺炎球菌、金黄色葡萄球菌和表皮葡萄球菌对青霉素、红霉素耐药率达50%以上。结论 本地区儿童呼吸道感染主要病原菌是革兰阴性杆菌,加强细菌培养和耐药性监测对临床合理使用抗生素有重要意义。     

大肠埃希菌和肺炎克雷伯菌对三种氨基糖苷类抗生素的耐药性分析

【摘 要】 目的 评价庆大霉素、妥布霉素及阿米卡星三种氨基糖苷类抗生素（AGs）对大肠埃希菌和肺炎克雷伯菌的体外抗菌活性。方法 对902株大肠埃希菌和404株肺炎克雷伯菌,采用VITEK-2全自动微生物分析仪配套的AST-GN13药敏卡进行庆大霉素、妥布霉素及阿米卡星的体外药敏试验。结果 大肠埃希菌和肺炎克雷伯菌的产超广谱β-内酰胺酶（ESBLs）菌株检出率分别为40.8％和36.6％;产ESBLs的大肠埃希菌对庆大霉素、妥布霉素及阿米卡星的敏感率分别为42.4%、39.1%和96.5%,与非产ESBLs菌株比较,敏感率差异均有统计学意义（P<0.01）;产ESBLs的肺炎克雷伯菌对庆大霉素、妥布霉素及阿米卡星的敏感率分别为64.2%、62.8%和91.9%,与非产ESBLs菌株比较,敏感率差异均有统计学意义（P<0.01）;阿米卡星对产与非产ESBLs的大肠埃希菌和肺炎克雷伯菌均高度敏感,敏感率均在91%以上。结论 本地区大肠埃希菌和肺炎克雷伯菌产ESBLs菌株流行严重,ESBLs的产生可使大肠埃希菌和肺炎克雷伯菌对AGs的耐药情况加重,提示ESBLs和AGs引起的耐药可能存在一定的相关性。