分子阵列研究

小分子化合物可以调节生物学过程,是研究活性生物大分子特别是蛋白质以及药物的重要工具,而高通量筛选是发现活性分子的重要方法.分子阵列是近年来新出现的一种高通量筛选技术,上面含有成千上万种组合合成的化合物以及天然产物,可以用于发现新的先导化合物,以及筛选已有的先导化合物.现在分子阵列已经成功应用于蛋白分析和先导化合物的发现等许多领域.本文综述了近年来分子阵列的构建过程、原位合成和非原位合成等各种固定化策略以及荧光免疫检测、表面等离子体共振成像技术等检测手段,并介绍了化学分子印刷阵列方法,最后总结了分子阵列的应用,并对分子阵列在我国中药发展等方面将起到的潜在作用作了展望.     

多样性导向的“类天然产物”化合物库合成

天然产物是药物发现中先导化合物的重要来源.高通量筛选技术的发展和近年来化学生物学研究的深入,对拓展天然产物与活性相关的＂化学空间＂提出了新要求.用多样性导向合成方法建立骨架多样、构造复杂、立体化学多样性的＂类天然产物＂化合物库进行生物学相关研究,并以此为基础发现药物先导化合物正在成为一种趋势.在此过程中,发展具有立体选择性和区域选择性,能够广泛应用于多种底物的有机化学反应起着关键作用.     

先导化合物的发现——整合计算机虚拟筛选、化学合成和生物测试方法

先导化合物的发现在药物研究中起关键作用。基于分子对接的虚拟筛选是创新药物研究的新方法和新技术,已成为一种与高通量筛选互补的方法,广泛应用于先导化合物的发现中。本文将结合本课题组的研究实例,综述了通过计算机虚拟筛选、化学合成和生物测试相结合的方法来发现先导化合物的一些研究工作。     

一种新的De Novo生物活性分子设计方法

由药效团进行虚拟活性结构生成与3D-QSAR模型相结合,筛选出有前途的结构多样性的化合物,并从中寻找活性先导化合物,是一种新的分子设计方法。采用这种方法对抗小麦赤霉病类含氟农药进行了研究,共生成了53个虚拟活性结构,通过3D-QSAR模型筛选出其中10个活性较高的结构,在活性最高的化合物基础上进行了结构修饰,得到了活性更高且毒性较低的理想化合物。研究结果表明这种方法能突破原模型化合物结构模式的局限,可以找到结构新颖的活性先导化合物,是一种非常有前途的分子设计方法,而且具有较高的筛选效率。     

临近闪烁分析法在高通量筛选中的应用

起源于放射性免疫分析的临近闪烁分析法(scintillation proximity assay,SPA)是一种均相、灵敏、快速和简便的基于闪烁载体的分析平台。该平台可用于筛选药物靶点的先导化合物和研究其生理过程。由于无需分离,易于固定药物靶点和检测其活性,SPA成为一种重要的高通量筛选方法。由于放射性标记分子和亲和标签分子的多样化和商业化、以及液闪计数器和液相操作等技术的发展,SPA已经广泛用于受体结合、高通量药物筛选、酶分析、放射性免疫分析、蛋白质-蛋白质相互作用和细胞水平分析等方面。本文阐述了SPA原理,讨论了其关键技术(包括闪烁载体、液闪计数器和放射性标记分子),分析了其评价体系;同时简述了SPA分析的发展, 并介绍了其在高通量筛选中的应用实例, 归纳了存在的问题,给出了未来的发展趋势。目前,基于SPA和荧光分析方法已成为高通量药物筛选的热点研究领域, 这些筛选技术的革新必然提升我们对细胞体系生物学的全面理解和促进先导化合物筛选过程的显著进步。     

药物发现中常用化合物库特征描述与对比分析

随着计算技术的发展和分子模拟软件的日趋成熟, 虚拟筛选已经在药物发现过程中发挥着越来越重要的作用. 在虚拟筛选过程中, 所使用化合物库的质量对先导化合物发现的成功率起着至关重要的作用. 本文通过对已知药物库、天然产物库、中药原植物化学成分库、筛选常用商业化合物库以及研究者所在实验室建立的化合物库的分析比较, 从化合物库的分子多样性、化学空间和分子骨架等多个方面提取并对比每一种化合物库的特征, 发现了已知药物库与中药原植物化学成分库的特征相似性, 揭示了中药原植物化学成分库作为筛选库的类药性优势, 并且深化了对几种筛选用化合物库特征的认识和理解.     

微波辅助组合合成的研究进展

微波辅助组合合成技术是近年来发展起来的一种新的制备化合物库的组合化学技术, 它不仅可以克服传统固相组合合成技术以及液相组合合成技术无法提高产物收率的不足, 而且利用该技术所制得的化合物库中对应的是高纯度的单一化合物, 采用高通量筛选技术可以快速直接地确定高活性结构, 极大地提高了新药开发的效率. 主要就近年来微波辅助组合合成技术的研究进展情况进行介绍, 内容包括固相组合合成、基于聚合物支载的催化剂的组合合成、液相组合合成、氟相组合合成以及组合平行合成等.     

质谱在高通量快速检测技术中的应用研究进展

质谱是现代分析测试技术中同时具备了灵敏度高、特异性好、分析速度快等特性的普适性方法,现已被广泛应用于食品检验、环境监测、药物筛选、临床医学等众多领域的高通量快速检测技术中。该文针对近年来低分辨质谱、高分辨质谱以及原位电离质谱技术在高通量快速检测,特别是小分子化合物的高通量快速检测中的应用研究进展进行了综述,并对质谱技术用于高通量快速检测的发展趋势及应用前景进行了展望,以期为相关领域的科技人员提供理论支持和技术参考。     

化学库及分子差异性设计研究进展

组合化学是近年来发展起来的一种快速合成和平行筛选群体化合物的新方法。它可以用来设计先导化合物，也可以对药物分子进行结构改造。本文主要综述了化学库的合成以及计算机辅助组合化学的研究进展。     

高温沉淀铁基催化剂上费托合成含氧化合物生成机理的研究

采用漫反射原位红外光谱法及化学捕获方法,考察了费托合成过程中高温沉淀铁基催化剂表面吸附物种的变化,并探讨了含氧化合物的生成机理。结果表明,CO在高温沉淀铁基催化剂上有线式和桥式两种吸附态存在,同时,CO的吸附在催化剂表面生成大量含氧化合物前驱体。费托原位实验捕获到了一些较关键的中间产物:表面乙酸盐、表面酰基、甲氧基等。同时发现高温沉淀铁基催化剂表面具有以下反应共性:醇类可与表面羟基结合生成烷氧基团;催化剂表面的吸附分子具有氧化性;一些基础化学物质如OH^-、晶格氧等可以与甲醇或乙醛分子发生反应。对CH₃OH + CO及CH₃I + CO + H₂两个反应的化学捕获实验表明,酰基是C₂⁺含氧化合物的重要中间产物,酰基加氢是形成C₂⁺含氧化合物的关键步骤。根据表面中间产物及反应特性,得到了高温沉淀铁基催化剂上费托合成含氧化合物的生成机理。     

Bioluminescence and chemiluminescence in drug screening

Drug screening, that is, the evaluation of the biological activity of candidate drug molecules, is a key step in the drug discovery and development process. In recent years, high-throughput screening assays have become indispensable for early stage drug discovery because of the developments in synthesis technologies, such as combinatorial chemistry and automated synthesis, and the discovery of an increasing number of new pharmacological targets.Bioluminescence and chemiluminescence represent suitable detection techniques for high-throughput screening because they allow rapid and sensitive detection of the analytes and can be applied to small-volume samples. In this paper we report on recent applications of bioluminescence and chemiluminescence in drug screening, both for in vitro and in vivo assays. Particular attention is devoted to the latest and most innovative bioluminescence and chemiluminescence-based technologies for drug screening, such as assays based on genetically modified cells, bioluminescence resonance energy transfer (BRET)-based assays, and in vivo imaging assays using transgenic animals or bioluminescent markers. The possible relevance of bioluminescence and chemiluminescence techniques in the future developments of high-throughput screening technologies is also discussed.     

Pharmacophore features of potential drugs

Drug discovery efforts rely increasingly on the identification of quality lead compounds through high-throughput synthesis and screening. However, large-scale random libraries have yielded only a low number of quality lead molecules. To address this shortcoming researchers have paid more attention to the concept of "drug-likeness" of molecules in combinatorial and screening libraries. Database profiling and analysis methods have been employed to identify the structural features of known drug molecules. Neural networks and machine learning methods help to distinguish between drugs and nondrugs. More recently, database-independent pharmacophore filters have been introduced that provide simple intuitive rules to classify potential drugs.     

Chemical protein synthesis-assisted high-throughput screening strategies for d-peptides in drug discovery

d-peptides are recognized as a new class of synthetic chemical drugs and they possess many interesting advantages such as high enzymatic stability, improved oral bioavailability, as well as high binding affinity and specificity. Recently, d-peptide drugs have been attracting increasing attention in both academic and industrial researches over recent years. One d-peptide etelcalcetide has even entered the market that targets the calcium (Ca²⁺)-sensing receptor (CaSR) to fight secondary hyperparathyroidism. Effective discovery and optimization of d-peptide ligands that can bind to various disease-related targets with high specificity and potency is of great importance for the development of d-peptide drugs. This review surveys the recent method development in this area especially the chemical protein synthesis-assisted high-throughput screening strategies for d-peptide ligands and their application in drug discovery.     

Towards the optimal screening collection: a synthesis strategy

The development of effective small-molecule probes and drugs entails at least three stages: 1) a discovery phase, often requiring the synthesis and screening of candidate compounds, 2) an optimization phase, requiring the synthesis and analysis of structural variants, 3) and a manufacturing phase, requiring the efficient, large-scale synthesis of the optimized probe or drug. Specialized project groups tend to undertake the individual activities without prior coordination; for example, contracted (outsourced) chemists may perform the first activity while in-house medicinal and process chemists perform the second and third development stages, respectively. The coordinated planning of these activities in advance of the first small-molecule screen tends not to be undertaken, and each project group can encounter a bottleneck that could, in principle, have been avoided with advance planning. Therefore, a challenge for synthetic chemistry is to develop a new kind of chemistry that yields a screening collection comprising small molecules that increase the probability of success in all three phases. Although this transformative chemistry remains elusive, progress is being made. Herein, we review a newly emerging strategy in diversity-oriented small-molecule synthesis that may have the potential to achieve these challenging goals.     

Small-molecule discovery from DNA-encoded chemical libraries

Researchers seeking to improve the efficiency and cost effectiveness of the bioactive small-molecule discovery process have recently embraced selection-based approaches, which in principle offer much higher throughput and simpler infrastructure requirements compared with traditional small-molecule screening methods. Since selection methods benefit greatly from an information-encoding molecule that can be readily amplified and decoded, several academic and industrial groups have turned to DNA as the basis for library encoding and, in some cases, library synthesis. The resulting DNA-encoded synthetic small-molecule libraries, integrated with the high sensitivity of PCR and the recent development of ultra high-throughput DNA sequencing technology, can be evaluated very rapidly for binding or bond formation with a target of interest while consuming minimal quantities of material and requiring only modest investments of time and equipment. In this tutorial review we describe the development of two classes of approaches for encoding chemical structures and reactivity with DNA: DNA-recorded library synthesis, in which encoding and library synthesis take place separately, and DNA-directed library synthesis, in which DNA both encodes and templates library synthesis. We also describe in vitro selection methods used to evaluate DNA-encoded libraries and summarize successful applications of these approaches to the discovery of bioactive small molecules and novel chemical reactivity.     

Application of Lectin Microarrays for Biomarker Discovery

Many proteins in living organisms are glycosylated. As their glycan patterns exhibit protein-, cell-, and tissue-specific heterogeneity, changes in the glycosylation levels could serve as useful indicators of various pathological and physiological states. Thus, the identification of glycoprotein biomarkers from specific changes in the glycan profiles of glycoproteins is a trending field. Lectin microarrays provide a new glycan analysis platform, which enables rapid and sensitive analysis of complex glycans without requiring the release of glycans from the protein. Recent developments in lectin microarray technology enable high-throughput analysis of glycans in complex biological samples. In this review, we will discuss the basic concepts and recent progress in lectin microarray technology, the application of lectin microarrays in biomarker discovery, and the challenges and future development of this technology. Given the tremendous technical advancements that have been made, lectin microarrays will become an indispensable tool for the discovery of glycoprotein biomarkers.     

高通量药物筛选现代检测技术研究进展

高通量药物筛选是发现创新药物的重要技术途径.高通量筛选结果必须通过适当的检测方法才能反映出来,检测技术是实现高通量药物筛选的基础.本文综述了近年来有关光学分析、色谱分析、热分析、电化学分析、质谱、核磁共振等现代检测技术在高通量药物筛选研究中的进展.     

Discovery,structure, and chemical synthesis of disulfide-rich peptide toxins and their analogs

Disulfide bond-rich peptide toxins are promising scaffolds for the development of medicinal peptides because they possess a rigid 3D structure formed by multiple disulfide bonds. In this review, we discussed recent advances in the discovery, structural elucidation and chemical synthesis of disulfide-rich peptide toxins and their analogs.     

Discovery of novel targets with high throughput RNA interference screening

High throughput technologies have the potential to affect all aspects of drug discovery. Considerable attention is paid to high throughput screening (HTS) for small molecule lead compounds. The identification of the targets that enter those HTS campaigns had been driven by basic research until the advent of genomics level data acquisition such as sequencing and gene expression microarrays. Large-scale profiling approaches (e.g., microarrays, protein analysis by mass spectrometry, and metabolite profiling) can yield vast quantities of data and important information. However, these approaches usually require painstaking in silico analysis and low-throughput basic wet-lab research to identify the function of a gene and validate the gene product as a potential therapeutic drug target. Functional genomic screening offers the promise of direct identification of genes involved in phenotypes of interest. In this review, RNA interference (RNAi) mediated loss-of-function screens will be discussed and as well as their utility in target identification. Some of the genes identified in these screens should produce similar phenotypes if their gene products are antagonized with drugs. With a carefully chosen phenotype, an understanding of the biology of RNAi and appreciation of the limitations of RNAi screening, there is great potential for the discovery of new drug targets.