Building Libraries with Chemical Diversity for Screening

我们与NIGMS资助的四个CMLD中的三个(波士顿大学、堪萨斯大学和匹兹堡大学)建立了材料转让协议(MTA)。这三个中心已经提供了10,000多个已知结构的样本。每个CMLD都有不同的库设计方法，因此它们的库之间的重叠可以忽略不计。MTL随后与其他学术和工业合作伙伴签订了许多协议，以扩大我们图书馆的广度。与传统药物化学小组的著名合作是与清华大学的刘刚教授和罗马大学的罗伯托·迪桑托教授。此外，我们还获得了一系列重点化合物，包括洛帕克、葛兰素史克Kline激酶文库、生物酚天然产物、ICCB已知生物活性物质，以及来自DCTD、NCI的一大系列合成化合物。作为美国国务院资助的一项减少威胁计划的一部分，哈萨克斯坦斯特普诺戈尔斯克前生物武器(BW)设施的科学家们被鼓励探索非BW科学视野。该计划是在哈萨克斯坦阿拉木图的国际科学技术中心(ISTC)的保护伞下进行的。MTL的工作人员曾四次前往哈萨克斯坦发展这一合作，最近收到了续签两年的通知。我们组织了以哈萨克斯坦特有的800种植物为重点的植物收藏。到目前为止，我们已经收集了177个植物样本；植物提取物已经在NCI 60细胞筛选中进行了测试，并在MTL筛选中进行了预分离。通过与两位韩国访问科学家Kim Young Ho Kim博士(韩国大田忠南国立大学)和Ik Soo Lee博士(韩国光州忠南国立大学)合作，我们获得了大量韩国植物提取物。我们与这些实验室的持续合作也使我们能够获得大量纯化的植物天然产品。耶鲁大学的Scott Strobel博士提供了一种独特的真菌内生菌提取物资源。这些内生菌是在一个本科生班级从秘鲁植物中培养出来的。学生们安排在耶鲁大学进行生物测试，然后分离出具有活性的天然产物。耶鲁大学的分子测试确保了培养的不同。本·申博士(佛罗里达州斯克里普斯)的实验室专门研究放线菌的分子遗传学，特别是非核糖体肽和聚酮合成酶系统。沈实验室还积极发现刺激隐秘生物合成基因表达的方法。通过与沈阳实验室的合作，我们已经获得了数百种具有明确遗传特征的放线菌提取物，以及一些纯化合物。从生物合成途径的遗传操作中获得的化合物在MTL筛选项目中提供了结构-活性信息。MTL最近收购了一家Matric微型商店，这是一种集成的机器人系统，用于存储和检索2D条形码管和板。自动化样品存储极大地提高了我们快速挑选HTS一次检测中发现的结果的能力，以确认活性和二次生物学评估。目前，该系统拥有180,703根管子和967块板材。在过去的4年里，CDDS开发了一个基于Web的定制化验和样本管理系统，使我们的研究人员能够管理样本库、设计化验试板以及存储和分析生物化验数据。

We established of Material Transfer Agreements (MTA) with three of the four NIGMS-funded CMLDs (Boston University, the University of Kansas, and the University of Pittsburgh). These three centers have provided over 10,000 samples of known structure. Each CMLD has a distinct approach to library design, thus there is negligible overlap between their libraries. The MTL has subsequently entered into numerous agreements with other academic and industrial partners to increase the breadth of our libraries. Notable collaborations with traditional medicinal chemistry groups are with Prof. Gang Liu of Tsinghua University, and Prof. Roberto DiSanto of the University of Rome. Furthermore, we have obtained a number of focused compound sets, including LOPAC, the Glaxo Smith Kline kinase library, BioMol Natural Products, ICCB Known Bioactives, as well as a large series of synthetic compounds from DCTD, NCI. As part of a US State Department-funded threat reduction program, scientists at the former bioweapons (BW) facility in Stepnogorsk, Kazakhstan have been encouraged to explore non-BW scientific horizons. The program is conducted under the umbrella of the International Science and Technology Center (ISTC), in Almaty, Kazakhstan. MTL staff traveled to Kazakhstan on four occasions to develop this collaboration, which recently received notice of renewal for two more years. We have organized plant collections focused on the 800 species endemic to Kazakhstan. To date, we have collected over 177 plant specimens; the plant extracts have been tested in the NCI 60 cell screen and prefractionated for testing in MTL screens. Through collaboration with two Korean visiting scientists, Dr. Young Ho Kim (Chungnam National University, Daejon, Korea) and Dr. Ik Soo Lee (Chonnam National University, Gwangju, Korea), we obtained an extensive collection of Korean plant extracts. Our ongoing collaboration with these laboratories has enabled us to receive a large collection of purified plant natural products as well. A unique resource of fungal endophyte extracts has been obtained from Dr. Scott Strobel at Yale University. The endophytes have been cultured from Peruvian plants in an undergraduate class. The students arrange for biological testing at Yale and then isolate active natural products. Molecular testing at Yale ensures that the cultures are distinct. The laboratory of Dr. Ben Shen (Scripps Florida) specializes in the molecular genetics of actinomycetes, particularly non-ribosomal peptide and polyketide synthase systems. The Shen lab is also active in discovering means for stimulating expression of cryptic biosynthetic genes. Through collaboration with the Shen laboratory we have obtained access to hundreds of actinomycete extracts with well defined genetics, and a number of pure compounds. Compounds obtained from genetic manipulation of the biosynthetic pathways have provided structure-activity information in an MTL screening project. The MTL recently acquired a Matrical MiniStore, an integrated robotic system for storing and retrieving both 2D bar coded tubes and plates. Automated sample storage has dramatically increased our ability to rapidly cherry pick hits found in HTS primary assays for confirmation of activity and for secondary biological evaluation. Currently the system holds 180,703 tubes and 967 plates. Over the past 4 years, the CDDS has developed a customized web based assay and sample management system which allows our researchers to manage sample libraries, design assay plates and store and analyze bioassay data.