KARI酶及其抑制剂

酮醇酸还原异构酶(keto-acid reductoisomerase, KARI或称作乙酰羟基酸异构还原酶(aceto hydroxyacid isomeroreductase, AHIR))是植物、微生物中的支链氨基酸生物合成途径中第二步的关键酶.由于哺乳动物和人体内不存在这种酶,因此靶向KARI酶的抑制剂具有安全、高效等特点,成为农用除草剂以及医药抗真菌药物的一个引人注目的研究方向.本文综述了KARI的酶学、KARI抑制剂及其抑制机理和理论研究进展,并对其发展趋势进行了展望.     

酮醇酸还原异构酶(KARI)与其抑制剂间相互作用的分子模拟研究

采用MCCE, Autodock及密度泛函方法对酮醇酸还原异构酶(KARI)与其抑制剂间相互作用进行了研究. 计算结果表明, KARI活性位点中的Mg^2＋在活性位点残基的离子化状态、活性位点的静电性质以及与抑制剂结合等方面起重要的作用; 同时, 抑制剂在结合方式、前线轨道布居及静电势等方面与酶促反应中间体(HOIV)具有一定程度的相似性; 可电离的羧基是当前发现的靶向KARI抑制剂一个重要的结构特征, 进一步推广可认为潜在的抑制剂应该拥有可电离成负电荷的功能团. 在药物设计中考虑到以上结论, 将有利于发现和改造靶向KARI的抑制剂.     

基于酮醇酸还原异构酶KARI复合物晶体结构的三维数据库搜寻

以酮醇酸还原异构酶KARI复合物0.16nm5高分辨率晶体结构为基础,采用DOCK 4.O分子对接程序通过MDDL/ACD三维数据库搜寻,找到了279个与KARI结合能较低的小分子,讨论了能量打分较高分子同靶酶的作用模式.这些分子作为潜在的KARI抑制剂为基于KARI的农药分子设计提供了有利指导.     

酰胺类KARI酶抑制剂的设计、合成和生物活性

以分子对接法进行MDL/ACD三维数据库搜寻所得到的大量结构信息为指导, 从中选取部分酰胺类小分子进行化学合成和结构表征. 生物活性测试结果表明, 设计合成的8个酰胺类化合物在200 μg/mL浓度下对水稻KARI酶具有不同程度的抑制活性, 化合物1和6的抑制率可达到57.4%和48.1%. 部分化合物在100 μg/mL浓度下对双子叶植物油菜的胚根和单子叶植物稗草的茎叶的抑制活性较为显著, 化合物1和6对油菜胚根生长抑制率分别为52.0%和72.6%, 其与KARI酶体外(in vitro)抑制活性具有一定的相关性. 这些酰胺类化合物可作为KARI酶抑制剂为后续分子设计提供借鉴.     

Molecular design,synthesis and biological activities of amidines as new ketol-acid reductoisomerase inhibitors

Diamidine （A） was identified in our in vitro bio-assay as a possible inhibitor of ketol-acid reductoisomerase （KARI） from the ACD database search based on the known three-dimensional crystal structure of KARI. An investigation on interaction of A on KARI active sites, led to the design and synthesis of 15 novel monoamidines. Some of those showed better biological activity than A on rice KARI （in vitro） and in greenhouse herbicidal tests （in vivo）. The structure-biological activity relationship was investigated, which provides valuable information to further study of potential KARI inhibitors.     

基于Ugi反应的含芳氨基甲酰基1-氰基-1-环丙烷甲酰胺类新化合物的合成、结构和生物活性研究

酮醇酸还原异构酶(KARI;EC 1.1.1.86)是植物和微生物(细菌、真菌)体内支链氨基酸生物合成过程中起催化作用的关键酶之一,可作为设计除草剂或杀菌剂的靶标.本工作在前期工作基础上,以1-氰基-1-环丙烷甲酸,芳基异腈,醛和胺为反应物,通过Ugi反应设计合成了一系列含芳氨基甲酰基1-氰基-1-环丙烷甲酰胺类新化合物8a~8p,经1H NMR,13C NMR,IR和元素分析或HRMS确证了结构,并解析了化合物8m的晶体结构.初步生物活性测试结果表明,这些环丙烷双酰胺类衍生物是结构新颖的KARI酶抑制剂,其中8a~8c,8m,8n和8j在200 mg/L浓度下对水稻KARI酶具有94%~98%的抑制活性,8m的Ki为(77.91±30.15)μmol/L.化合物在50 mg/L浓度下对黄瓜枯萎病菌、花生褐斑病菌、苹果轮纹病菌、番茄早疫病菌和小麦赤霉病菌等5种植物病菌表现出显著抑制活性,整体来看8e和8p具有相对宽泛的抑菌谱,可作为新型抑菌剂苗头化合物进行深入研究.     

脒类KARI酶抑制剂的分子对接和3D-QSAR研究

对设计合成的20个单脒类化合物的水稻KARI酶体外抑制活性和体内除草活性进行了分子对接和三维定量构效关系研究. 前者采用AutoDock3.0方法, 研究发现化合物活性变化趋势与分子对接计算结果基本一致, 通过分析化合物9与KARI酶活性氨基酸残基结合模式发现, 残基Glu319, Asp315, Glu496, Gly253, Met254, Cys517等对氢键和静电相互作用以及疏水作用都有重要贡献; 后者研究采用比较分子力场分析(CoMFA)方法, 结果表明立体场和静电场对活性的贡献分别为67.8%和32.2%, 交叉验证系数r_cv²＝0.774, 非交叉验证r²＝0.999, F＝1593.134, 标准偏差s＝0.036, 所建立的3D-QSAR模型对化合物除草活性具有较好的预测能力. 两种方法研究结果为进一步设计合成更高活性的KARI酶抑制剂提供了指导.     

Synthesis and Crystal Structure of 1-[4-(Pyrimidin-2-yl)piperazin-1-ylmethyl]-1H-benzotriazole

1 INTRODUCTION Based on the reported 1.65 ? high resolution crystal structure of spinach KARI (ketol-acid reduc- toisomerase) complex[1], we obtained 279 molecules with low binding energy toward KARI from MDL/ ACD 3D database searching, using program DOCK 4.0[2]. These potential structures provide further information for the design of new KARI inhibitors, one of novel derivatives of which as the title com- pound has been synthesized. Its crystal structure will provide us more inf…     

新型N-（4，6-双取代嘧啶-2-基）-N’-（三氟甲基苯基）胍的合成，结构及生物活性研究

设计,合成了10种新型的胍衍生物,结构经FTIR, MS, 1H NMR 和元素分析确证,并且采用X-射线衍射分析方法确证了具有较好生物活性的化合物11a的结构。并且对这些化合物进行了除草活性测试,结果表明化合物11a,11b,11c在100µg·mL-1 对油菜具有较好的抑制作用,初步的KARI活性结果表明这些化合物对KARI的活性很弱。     

环肽类组蛋白去乙酰化酶抑制剂

组蛋白去乙酰化酶抑制剂是一类以组蛋白去乙酰化酶为靶点的新型靶向抗肿瘤药物,在抗增殖、促凋亡、促分化、阻滞细胞生长周期、抗血管生成等方面有很好的作用。组蛋白去乙酰化酶抑制剂以其独特的抗肿瘤作用机理,在研发肿瘤治疗药物中占有重要地位。在组蛋白去乙酰化酶抑制剂分类中,环肽类抑制剂结构最为复杂,对多种类型的实体瘤及血液学癌细胞均具有良好的对抗作用。本文针对天然和化学合成的环肽类组蛋白去乙酰化酶抑制剂中金属结合区、表面识别区以及连接区的结构特点进行了综述,并描述了各类抑制剂对酶的抑制活性和抗肿瘤增殖活性。对抑制剂不同结构区的修饰、改造可以使抑制剂对不同肿瘤细胞具有高效性和特异性作用,通过构效关系研究寻找具有高效低毒、靶向性环肽类抑制剂的结构规律,可为研究开发抗肿瘤药物提供帮助。     

可录型光盘与菁染料光存储媒体的研究进展

本文简介绍了可录型光盘光学数据存储过程和作为存储介质同的菁染料５的稳定性及其应用研究概况,学一些新型光存储技术作了简要评述,并讨论了可录型光存储材料的发展趋势。     

AFM analysis of CD-R photoageing

One of the main parameters controlling durability of optical discs such as CD-R is the stability of the dye layer. Ambient light exposure contributes to degradation of the light-sensitive dye used in recordable media. Our study focuses how light irradiation influences the dye layer on a polycarbonate (PC) substrate before and after CD-R engraving. The dye layer surface was characterized by atomic force microscopy (AFM) to measure dye degradation, which, in turn, can be used to assess CD-R readability. For a blank CD-R, pre-groove height is a major degradation indicator. For a recorded CD-R, however, pit and land size and depth can be used to show degradation. For long irradiation times, coded areas completely disappear, leading to the loss of stored information.     

Current developments in optical data storage with organic dyes

The main motivation for the development of digital data storage has been the improvement in play‐back quality and the increase in storage capacity. In 1982 Philips and Sony introduced the first technically and economically successful system based on this—the compact disc (CD) and a compatible player. A very broad diversity of optical data recording formats are available today, and a difference is drawn between prerecorded, recordable, and rewritable media. This Review gives an overview of the systems used, the main features of production, and then concentrates on the properties of the organic dyes that are used in recordable systems. Dyestuffs chemistry has gained the reputation of having become a mature field of activity. Is this prejudice or a justified swan song for dyestuffs chemistry? When applications in optical data storage are considered, it is evident that even today progresses such as CD‐R and DVD/R would not be feasible without functional dyes.     

高密度可录光盘存储材料:金属卟啉配合物*

金属卟啉配合物在400-650 nm波段具有优良的光学性质,是一类新型高密度可录光盘存储介质材料.本文在总结近年来最新研究成果的基础上,阐述了金属卟啉配合物的结构特点、合成方法和作为高密度可录光盘存储介质的作用机理,讨论了影响卟啉配合物性质和光存储性能的主要因素,并对未来的发展趋势作了展望.     

利用AFM研究溶剂对菁染料薄膜光学性能的影响

在CD-R光盘中,要求记录介质薄膜有良好的光学性能,这样才能降低噪音,提高数据存储与读取的可靠性.菁染料是应用最广泛的记录介质,本文采用旋涂法制备菁染料薄膜时,发现溶剂对薄膜的光学性能有很大的影响.通过AFM对薄膜表面的微观精细结构的研究表明,染料薄膜反射率的高低与薄膜表面粗糙度有关.其中使用氯仿、四氟丙醇和二丙酮醇作溶剂时,制备的染料薄膜表面平整,反射率较高;使用甲醇作溶剂时,染料薄膜布满直径0.3μm左右的晶粒,对光线有强烈散射作用,从而降低了染料薄膜的光学性能.     

Disposable twin gold electrodes for amperometric detection in capillary electrophoresis

We describe a simple and easy way to construct gold microelectrodes for amperometric detection in capillary electrophoresis (CE). The gold microelectrodes, in single or twin sets, were obtained from recordable compact discs (gold-sputtered type), which present highly reproducible surface characteristics. The performance of these electrodes was evaluated by using a home-made CE equipment. The basic steps for the electrode construction are: drawing on a microcomputer; laser printing of the design on wax paper; heat-transfer of the toner onto the gold surface of a peeled recordable compact disc (CD-R); etching of the gold layer from unprinted regions; removal of the toner with a solvent; sealing of unused electrode areas with varnish. One electrode at a time was connected to a potentiostat (or two, to a bipotentiostat) and operated in a wall jet configuration relative to the CE capillary outlet. The amperometric signals were integrated for quantification purposes. Repetitive injections (n = 10) of a mixture containing iodide, ascorbic acid, dipyrone, and acetaminophen (20, 200, 500, and 100 microM), presented relative standard deviations of 2.9, 4.5, 6.1, and 4.0%, respectively. For these analytes, the detection limits (S/N = 3, 30 s of 100 mm hydrodynamic injection) were 0.1, 0.5, 3.1, and 1.1 microM, respectively.     

Warpage prediction of optical media

The warpage of injection–compression‐molded optical media, such as compact discs and digital video discs, due to asymmetric cooling during production is predicted. Thermally induced stress is calculated with a nonisothermal compressible flow simulation with a viscoelastic constitutive model. A finite element analysis is formulated with axisymmetric plate elements based on Kirchhoff thin‐plate theory to simulate the warpage of the disc due to the asymmetric thermal stress and gravity after demolding. Simulation results of warpage for compact‐disc‐recordable moldings are compared with experimental observations under different processing conditions, such as the melt temperature, mold temperature, and packing pressure, with an optical grade of polycarbonate. The comparison shows that the simulation well predicts the effects of various processing conditions. Both the simulation and experiment indicate that of the processing conditions, the mold temperature has the greatest effect on warpage. © 2003 Wiley Periodicals, Inc. J Polym Sci Part B: Polym Phys 41: 859–872, 2003     

Materials for Optical Data Storage

The outstanding feature of materials research in the eighties has been the convergence of basic research and practical application, leading to ever shorter cycles of innovation. This is especially true of materials which form the basis of key technologies. The mass storage units of the next generations of computers will be based on optical processes having a storage density which exceeds that of all hitherto known storage techniques that are practicable from the technical standpoint. In view of the fact that since 1982 read-only memories in the form of compact discs (CD-ROM) have become very successful in the field of audio electronics, research and development are now concentrated on materials for write-once (WORM) and erasable (EDRAW) storage systems. Suitable materials for optical data storage are substances in which data markings can be recorded and deleted respectively using semiconductor lasers. Materials development is centered on the synthesis of infrared-absorbing dyes for WORM memories and the production of rare earth/transition metal alloys for magneto-optical data recording. An introduction to CD-ROM technology will be followed by an overview of the state of development of the most important storage materials which are currently available commercially, and then the properties of these materials will be discussed with reference to selected examples.     

聚合物光盘基板材料的研究进展

光盘是一种以非接触方式记录/再现信息的信息记录介质,主要由基板、记录层和保护层组成。其中,基板材料所用的光学塑料是光盘生产的主要原材料,对光盘的性能和成本构成至关重要。本文综述了聚合物光盘基板材料的研究现状及其发展趋势。光盘基板材料主要包括传统的聚碳酸酯（PC）,聚甲基丙烯酸甲酯（PMMA）、环氧树脂以及近年发展起来的非晶环烯烃聚合物。研制具有较低的双折射、吸湿性、固化成型收缩率以及较高的存储密度和存储寿命的光盘基板材料是聚合物光盘基板材料的一个重要研究领域。     

Light-Driven Reversible Multicolor Supramolecular Shuttle

Light-driven multicolor supramolecular systems mainly rely on the doping of dyes or a photo-reaction to produce unidirectional luminescence. Herein, we use visible light to drive the bidirectional reversible multicolor supramolecular shuttle from blue to green, white, yellow, up to orange by simple encapsulation of spiropyran-modified cyanostilbene (BCNMC) by the macrocyclic cucurbit[8]uril (CB[8]) monomer. The resultant host–guest complex displayed enhanced fluorescence properties, i.e. the multicolor fluorescence shuttle changed from blue to orange in the dark within 2 hours and reverted to the original state upon visible light irradiation for 30 s. Benefiting from the sensitivity of the spiropyran moiety to light, it can spontaneously isomerize from the ring-opened state to a ring-closed isomer in aqueous solution, and this photo-isomerization reaction is a reversible process under visible light irradiation, leading to the multicolor luminescence supramolecular shuttle as a result of intramolecular energy transfer. In addition, the light also drove the reversible conversion of the topological morphology of the host–guest complex from two-dimensional nanoplatelets to nanospheres. Different from the widely reported molecular rotaxane “shuttle”, the spiropyran supramolecular shuttle confined in the macrocyclic host CB[8] not only modulated a reversible topological morphology by light but also exhibited multicolor luminescence, which was successfully applied in programmed and rewritable information encryption.