非标记小分子药物靶点发现技术及其在中药研究中的应用

药物靶点的发现在药物研发中发挥着至关重要的作用,非标记的药物靶点发现技术因其无需改造药物小分子结构,能更真实反映小分子药物与靶点互作,成为药物靶点发现的新途径。中药(traditional Chinese medicines,TCMs)具有多成分、多靶点的特性,其作用靶点及机制的研究备受制约,鉴于非标记的药物靶点发现技术在TCMs化学成分靶点发现研究中的应用潜力,本文综述了非标记的药物靶点发现新技术及其在TCMs研究中的应用,为TCMs化学成分靶点的发现提供参考,为促进TCMs现代化的发展提供新思路。     

多靶点药物治疗及药物发现

作用于单靶点的药物在治疗复杂性疾病如肿瘤、糖尿病、感染性疾病时常常疗效不佳或毒性较大。多靶点药物可以同时调节疾病网络系统中的多个环节,不易产生抗药性,对各靶点的作用产生协同效应,达到最佳的治疗效果。本文对多靶点药物治疗的特点、分类情况、发现策略、筛选模型及已在临床使用的多靶点治疗药物进行综述,并探讨中药在多靶点药物治疗应用中的潜力。     

基于全球上市药物数据的抗阿尔茨海默病重定位新药发现

目的从上市药物中寻找潜在的多靶点抗阿尔茨海默病(AD)药物。方法本文利用实验室前期基于机器学习算法建立的抗AD多靶点药物预测平台,对全球上市药物数据库进行了预测和挖掘。结果从全球上市药物数据库中预测出13个至少作用于1个抗AD药物靶点,且可以通过多种作用对抗AD的上市药物,利用Cytoscape构建化合物——靶点网络;并针对上市药物阿戈美拉汀及其预测的多靶点蛋白进行分子对接以验证预测结果,蛋白靶点包括ADORA2A,ACHE,BACE1,PTGS2,MAOB,SIGMAR1和ESR1,阿戈美拉汀均能与以上靶点产生相互作用。结论本研究应用机器学习算法、网络药理学方法及分子对接方法,初步揭示了上市药物数据库中抗AD作用的潜在药物,为上市药物抗AD作用重定位提供了重要信息。     

基于HDAC的双靶点抗肿瘤药物研究进展

多靶点作用的抗肿瘤药物比目前单靶点药物具有更好的药效,且能够降低耐药性和毒副作用。为了探索多靶点药物在肿瘤化疗中的应用前景,以组蛋白去乙酰化酶(HDAC)抑制剂为基础设计多种作用的双靶点抑制剂已经成为了研究热点,其中部分化合物抑制肿瘤细胞增殖活性比现有的上市药物更好。本文综述了基于HDAC的双靶点抑制剂的研究进展,重点介绍了作用机制、设计策略和生物活性。     

药物靶点的发展前景及需要解决的关键问题

[摘要]随着人类基因组计划的完成、基因功能的发现,以及各种高通量筛选方法的实现,药物靶点已经成为了现代创新药物研发的一个重要环节。但进入临床试验和上市的新药数量并未随着潜在靶点的增加而有所增长,甚至还出现了下降趋势。文中分析了目前创新药物数量减少的原因,阐明了科学技术发展进程对药物靶点的影响,在此基础上预测了药物靶点的发展前景;同时,也对药物靶点领域需解决的关键问题进行了探讨。     

基于网络药理学方法探讨犀角地黄汤治疗系统性红斑狼疮的作用机制（英文）

本研究通过网络药理学方法,探讨犀角地黄汤治疗系统性红斑狼疮(SLE)的作用机制。首先通过TCMSP、TCMID和BATMAN-TCM数据库检索犀角地黄汤各中药成分的活性成分及其靶点,汇总得到药物靶点;其次在CTD、OMIM和TCMIP数据库检索得到SLE的疾病靶点,并对药物靶点和疾病靶点的交集进行分析。利用Cytoscape3.7.1软件构建药物-成分-靶点-疾病网络。在String平台构建治疗靶点的蛋白质互作网络(PPI),筛选其关键模块和Hub基因,在DAVID数据库进行GO和KEGG富集分析。最终筛选出22个犀角地黄汤的活性成分, 209个药物靶点, 284个疾病靶点, 59个治疗靶点。药物-成分-靶点-疾病网络和PPI网络分析得到槲皮素、山奈酚、黄芩素等核心活性成分, PTGS2、CASP3、MMP9、AKT1、JUN、CXCL8、FOS、TP53等核心靶点。主要涉及TNF、PI3K-Akt、Toll样受体、T细胞受体、细胞凋亡等信号通路。研究表明犀角地黄汤可能通过调控炎症、免疫及细胞凋亡等信号通路,发挥抗炎、抑制异常免疫应答和细胞凋亡的作用,具有多中心、多靶点、多途径的特点。     

多靶点鬼臼毒素新衍生物CCP-1抗肿瘤多药耐药作用机制研究

<正>以往单一靶点的抗肿瘤药物极易产生耐药[1-3]。多靶点的化疗药物具有其独特优势,如多靶点的化疗药物是单分子化合物,不存在多药物联合应用中的药物相互作用问题,也可避免药物联合应用中药物配伍、剂量、给药方式等问题。本课题组以天然产物中鬼臼毒素为母核,整合了具有抗肿瘤耐药的小分子片段,将鬼臼毒素新衍生物设计为同时作用于多个耐药靶点的化合物,以更好的克服传统单靶点化疗药物     

多靶点作用药物及其设计

疾病起因的多因素性导致单靶点作用药物活性的降低或副作用的增加,利用多靶点作用的药物有可能有利于解决这些问题,本文就多靶点作用药物设计所存在的问题和解决方法进行探讨,提出利用药效团模型法设计多靶点作用药物的理念。     

化学蛋白质组学与药物靶点的发现

小分子药物靶点的发现对于生物和医学的研究者而言,是一项既重要又艰巨的任务,医学和药学界研究工作者急切需要发现和确认新的靶点。为了克服药物靶点确认的瓶颈,已经发展了许多新技术用以研究小分子化合物与蛋白质分子间的相互作用,其中包括化学蛋白质组学方法。化学蛋白质组是全蛋白质组学研究的一个亚类,化学蛋白质组学是利用能够与靶蛋白质发生特异性相互作用的化学小分子来干扰和探测蛋白质组,在分子水平上系统揭示特定蛋白质的功能以及蛋白质与化学小分子的相互作用,从而准确找到药物作用靶点的组学研究方法。化学蛋白质组学技术和方法不断成熟,在药物作用靶点的发现、确认和药物多靶点研究等方面都将起到重要的作用,并将大大提高药物发现的效率。     

多靶点药物分子设计

作用于单一分子靶标的药物治愈多基因相关疾病如癌症、或影响多个组织或细胞类型的疾病如糖尿病等存在的问题逐渐被人们所认识。与选择性药物的治疗作用相比,几个靶标间的平衡调节能够提供较好的疗效和较低的副作用,同时作用于多个靶标的多靶点药物能够较好地控制复杂的疾病。本文详细比较分析了单靶点药物的不足和多靶点药物的优势,介绍了多靶点配体药物分子设计的方法及需要优化的参数。对于多靶点药物设计,关键的挑战是如何保证获得平衡的活性同时又能够实现选择性以及适当的药代动力学性质。到目前为止, 多靶点药物分子设计的方法对于药物化学家、药理学家、毒理学家以及生物化学家仍然是一项新的挑战。     

Recent developments in C5/C5a inhibitors

Complement factor 5a (C5a) is formed upon complement system activation in response to infection, injury or disease. Whilst C5a is a potent mediator of immune and inflammatory processes, excessive production or inadequate regulation of C5a has been implicated in the pathogenesis of numerous immuno-inflammatory diseases, predominantly through experimental studies utilising animal models of disease. Both acute and chronic conditions may benefit from C5a inhibition, including rheumatoid arthritis, inflammatory bowel disease, asthma, psoriasis, haemorrhagic shock and neurodegenerative conditions. The potentially broad clinical application for treatments that inhibit the activity of C5a at C5a receptors and the large global market for anti-inflammatory therapeutics have made C5a and the C5a receptor attractive targets for academic and commercial drug development programmes. In the past 5 years, interest in C5a as a drug target has grown substantially, and this activity has resulted in a collection of patents and scientific papers reporting novel C5a and C5a receptor inhibitors and antagonists, and generated a secondary stream of patent applications broadly claiming the use of C5/C5a inhibitors as a method of treating various immune and inflammatory conditions. This paper will review the physiology and pathophysiology of C5a and discuss the development of C5a and C5a receptor inhibitors in light of the recent scientific and patent literature.     

Effect of carbamazepine on expression of UDP-glucuronosyltransferase 1A6 and 1A7 in rat brain

Because UDP-glucuronosyltransferase (Ugt) 1a6 and Ugt1a7 are highly expressed in the rat brain, changes in Ugt1a6 and Ugt1a7 expression may affect the pharmacokinetics of drugs and endogenous compounds in the brain. The present study aimed to elucidate the effect of carbamazepine (CBZ), a typical UGT inducer, on Ugt1a6 and Ugt1a7 expression in the rat brain. Sprague-Dawley rats were treated intraperitoneally for 7 d with CBZ (100 mg/kg/d). Ugt1a6 and Ugt1a7 mRNAs were induced by CBZ in the cerebellum, piriform cortex, and hippocampus (Ugt1a6: 3.1-, 2.4-, and 1.9-fold, respectively, Ugt1a7: 2.3-, 1.6-, and 3.1-fold, respectively); serotonin glucuronidation, which is catalyzed by Ugt1a6, was also increased by 2.8-, 1.7-, and 1.8-fold in these regions, respectively. The nuclear translocation of the constitutive androstane receptor was increased 1.4-fold in the cerebellum and piriform cortex, suggesting that brain Ugt1a6 and Ugt1a7 might be induced via the constitutive androstane receptor. However, the pregnane X receptor and nuclear factor erythroid 2-related factor 2 did not play decisive roles in the induction. Histone H3 lysine 9 acetylation, H3 lysine 4 pan-methylation, and H3 lysine 9 mono-methylation may not be required for the induction. This study clarified that CBZ affected Ugt1a6 and Ugt1a7 in the brain.     

微小 RNA-20a、微小 RNA-92a在乳腺癌诊断及预后评估中的应用

目的 分析微小RNA(miR)-20a、miR-92a在乳腺癌病人血清中的表达情况,并探讨其对乳腺癌诊断及预后评估的价值.方法 选取2012年1月至2013年5月中国人民解放军中部战区总医院首次确诊为乳腺癌并经手术治疗的病人89例为观察组,以80例同期健康体检者为健康对照组,采用实时荧光定量逆转录聚合酶链式反应(qRT-PCR)检测乳腺癌病人术前和术后及健康对照组血清miR-20a和miR-92a表达水平,分析miR-20a和miR-92a表达与临床病理特征的关系.Spearman相关性分析血清miR-20a和miR-92a表达相关性,ROC曲线分析miR-20a、miR-92a以及二者联合检测对乳腺癌的诊断价值,Kaplan-Meier法分析miR-20a和miR-92a表达对病人5年总生存期(OS)影响,logistic回归分析影响乳腺癌预后的因素.结果 乳腺癌病人血清miR-20a[术前(1.76±0.15)、术后(1.38±0.12)比(1.02±0.08)]和miR-92a水平[术前(1.83±0.13)、术后(1.45±0.11)比(1.11±0.07)]较健康对照组均显著升高(P<0.05).miR-20a和miR-92a表达与病人肿瘤分化程度、TNM分期及淋巴结转移相关(P<0.05).乳腺癌病人血清miR-20a和miR-92a表达呈线性正相关(r=0.52,P<0.05).miR-20a和miR-92a二者联合诊断乳腺癌的敏感度、特异度以及AUC均显著升高(P<0.05).miR-20a和miR-92a低表达病人5年OS均显著高于高表达病人[miR-20a(93.24％比73.28％)、miR-92a(94.85％比74.57％),P<0.05].miR-20a和miR-92a、分化程度、淋巴结转移均是影响乳腺癌病人预后的独立危险因素(P<0.05).结论 乳腺癌病人血清中miR-20a和miR-92a表达异常,与病人肿瘤分化程度、TNM分期及淋巴结转移相关,可能用于乳腺癌的诊断和预后判断.     

Average Bioequivalence Evaluation: General Methods for Pilot Trials

In clinical development of a bioequivalent (BE) drug product, a two-step strategy is commonly adopted. In the first step, a pilot BE trial is conducted to evaluate the acceptability of the test drug product as a candidate for further evaluation in a subsequent pivotal BE trial. In the second step, a full-scale pivotal BE trial is conducted to formally establish bioequivalence. The objective and criterion of a pilot BE trial are different from those of a pivotal BE trial. In practice, however, a pilot BE trial is often inappropriately designed and analyzed based on the criterion for a pivotal BE trial. One main reason is the lack of well-established design and analysis methods for a pilot BE trial. To close this gap in practice, this study proposes a Pilot Acceptance Range method specifically constructed for analyzing a pilot BE trial within the framework of a two-step strategy. For designing a crossover pilot BE trial, this paper derives the power function and provides an easy-to-use method for determining the sample size.     

miR-422a靶向激肽释放酶-4抑制宫颈癌细胞的增殖、迁移与侵袭

目的 探讨miR-422a靶向激肽释放酶-4（KLK4）对宫颈癌细胞增殖、迁移和侵袭的影响。方法 实时荧 光定量PCR法（qPCR）检测人宫颈癌细胞HeLa、SiHa和人正常宫颈上皮细胞H8中miR-422a的表达;将宫颈癌HeLa 细胞分为 blank 组、miR-NC 组、miR-422a 组、mut miR-422a 组、miR-422a+pcDNA 组、miR-422a+pcDNA-KLK4 组。 CCK-8实验和Transwell实验检测各组宫颈癌HeLa细胞增殖、迁移和侵袭的能力;TargetScan软件预测miR-422a可 能调控结合的靶基因,双荧光素酶活性实验对靶向关系进行验证。Western blot检测各组细胞KLK4蛋白表达水平。 结果 与人正常宫颈上皮细胞H8相比,宫颈癌HeLa、SiHa细胞中miR-422a呈现低表达（P＜0.01）,选择宫颈癌HeLa 细胞进行后续实验;与miR-NC组和blank组相比,miR-422a组培养4、6 d细胞增殖能力降低,培养6、12 h细胞迁移 和侵袭数量减少（P＜0.05）;与miR-422a+pcDNA组相比,miR-422a+pcDNA-KLK4组培养4、6 d细胞增殖能力明显 升高,培养6、12 h细胞迁移和侵袭数量增加（P＜0.01）。TargetScan软件预测发现KLK4基因与miR-422a存在结合位 点,并经双荧光素酶报告基因实验验证。与 miR-NC 组相比,miR-422a 组中 KLK4 蛋白的表达水平降低,与 miR- 422a 组相比,mut miR-422a 组中 KLK4 蛋白的表达水平升高（P＜0.01）,与 miR-422a+pcDNA 组相比,miR-422a+ pcDNA-KLK4组中KLK4蛋白的表达水平升高（P＜0.01）。结论 miR-422a可通过靶向KLK4蛋白的表达抑制宫颈 癌细胞增殖、迁移和侵袭。     

EVALUATING QUALITATIVE ASSAYS USING SENSITIVITY AND SPECIFICITY

Sensitivity and specificity are two important indices of performance of qualitative assays. Evaluating these indices usually requires one to identify the true disease state of each subject involved in a study. This implies that a perfect test, a “gold standard,” is needed to test each subject. However, a gold standard test cannot always be performed on all subjects, whether because of cost or adverse effect on a subject's welfare. In these situations, a common practice is to apply both a currently used assay and an investigational assay to the same specimen. If the testing results are discordant, a gold standard test is applied. This approach has been criticized by many and, in fact, the statistics based on this approach usually overestimate sensitivity and specificity. This paper proposes two alternative methods to estimate sensitivity and specificity. Simulation results show that these methods perform better than the commonly used existing ones.

This paper proposes new acceptance criteria and designs to specific topics for the evaluation of blood related assays as well. To evaluate a qualitative assay related to blood specimens, one must also perform studies of storage conditions, interfering substances, and other related factors, in order to establish the equivalency of the assay under standard and various other conditions. To conduct these studies, true negative blood donor specimens are used as a sample from a nondiseased population; and blood donor specimens with spiked analyte are used to represent a sample from a diseased population. Currently, the target-spiking ranges and sample sizes are determined subjectively. This paper presents new acceptance criteria on acceptable conditions and objective standards for selecting the target-spiking range and sample size.     

Distribution of Phospholipids and Triglycerides in Multivesicular Lipid Particles

The distribution of phospholipid and triglyceride molecules in themembranes forming the nonconcentric vesicular network within a multivesicular lipid particle (MLP) and its precursors was investigated. MLP formulations afford controlled release of encapsulated pharmaceuticals over a time period ranging from a few days to a few weeks. The formation of MLP requires the use of a double emulsification process and a neutral lipid such as a triglyceride. MLP formulations with the antineoplastic agent cytarabine encapsulated in the aqueous compartments were prepared that further contained [13C]carbonyl-enriched triolein, and quantitative 13C nuclear magnetic resonance (NMR) experiments were performed to quantitate membrane-soluble and bulk triglyceride. In the NMR spectra of MLP, components attributable to triolein residing in a location similar to that of bilayer phospholipid were not observed. Spectral components attributable to triolein in a liquid-like phase were observed, and the intensity of these components accounted for the triolein present in the sample. Laser scanning confocal microscopy with two phospholipid fluorescent probes, RhodamineDHPE and NBD-PG, and a triglyceride fluorescent probe, BodipyTriglyceride, was used to visualize the distribution of the lipid components in the emulsion precursors of MLP and in MLP themselves with cytarabine, amikacin, morphine, or sucrose encapsulated within the aqueous phases. The confocal micrographs revealed a uniform distribution of the two phospholipid probes in the plane of the membrane, whereas the triglyceride probe accumulated at discrete locations. The NMR and microscopy results are consistent with a structural model for MLP in which triolein acts as a hydrophobic space filler at bilayer intersection points and stabilizes these junctions, and is also present as oil droplets dispersed in the encapsulated aqueous compartments. The experimental approach presented here could provide a rational approach to the development of MLP formulations with variable rates of sustained release, modulated by changes in the distribution of various phospholipids and triglycerides.     

Centralized preparation of hazardous drugs

In 1987, the manager of the Saint-Joseph Hospital (Paris, France) requested a reorganization of cytotoxic drug preparation. Protection for staff who handle hazardous drugs was the main concern. The conclusions drawn from a first analysis emphasize the advantages of a centralized reconstitution unit against a decentralized system. Subsequently, a workload study and an economic study (investment, maintenance, supplies, staff costs, comparative balance sheet and a 5-year simulation) were carried out, but to choose between a laminar airflow in aseptic room and an isolator in a conventional room. The selected isolator is the first of the conception: the central half-suit uses as a server, and four sleeves located on one side allow two technicians to work in a sterile and closed area without sterile garments.     

输尿管镜术中保留斑马导丝对减少输尿管损伤的作用研究

目的探讨在输尿管镜手术中保留斑马导丝对减少手术并发症和保证手术顺利进行的作用。方法将240例输尿管结石患者随机分为观察组及对照组,各120例,均在斑马导丝引导下进镜：其中观察组在输尿管镜到达结石部位后斑马导丝越过结石并保留于输尿管内,退镜后重新进镜至结石处并用钬激光碎石取石,术后留置双J管;对照组则在输尿管镜到达结石部位后退出斑马导丝,直接用钬激光碎石取石,术后留置双J管;术后3～5d复查腹部x线平片（KUB）和B超,观察两组疗效,对手术成功率、结石移入肾盂率、输尿管腔道丢失率和输尿管损伤率进行对比分析。结果在手术成功率方面：观察组120例均取得成功,成功率100．0％（120／120）;对照组100例取得成功,成功率83．3％（100／120）;两组间差异具有统计学意义（P〈0．05）。结石移入肾盂方面：观察组上段有14例移入肾盂,上移率25％（14／56）,中下段4例移入肾盂,上移率6．25％（4／64）;对照组上段24例移入肾盂,上移率48％（24／50）,中下段9例移入肾盂,上移率12．9％（9／70）;两组间差异具有统计学意义（P〈0．05）。输尿管腔道丢失率方面：观察组无病例出现腔道丢失,腔道丢失率0．0％;对照组15例出现腔道丢失,腔道丢失率12．5％（15／120）;两组间差异具有统计学意义（P〈0．05）。输尿管损伤率方面：观察组4例发生输尿管损伤,损伤率3．3％（4／120）;对照组14例发生输尿管损伤,损伤率11．7％（14／120X两组间差异具有统计学意义（P〈0．05）。结论在输尿管镜手术中保留斑马导丝能有效减少手术并发症和保证手术顺利进行,使手术更安全、有效。