NCI Program for Natural Products Discovery - Cures

1. 总体进展a.列出计划的预期目标和结果。目的1。创建新技术，建立一个增强的NP预分馏库，适用于现代高通量靶向筛选程序。目标2。利用新的方法和文库扩大可培养微生物的化学多样性。目标3。为世界各地的筛选中心提供预分离库，加速药物发现。目标4。为研究人员提供高通量筛选支持，以实现有针对性的发现工作。目标5。提供更快的分析资源（隔离、结构解析、再供应）以加快转化过程。目标6。建立公共数据库和生物信息学平台，扩大投入，扩大影响。b.总结报告期（FY22）取得的进展。1. 在2022财年，NPNPD达到了18,000个土壤真菌的水平，这些真菌被镀，拍照，并冷冻瓶长期使用。此外，还进行了410次大规模发酵和提取。2. 完成了1000种海洋放线菌和1000种海洋真菌的基因测序和分类鉴定（也正在进行镀片、照相和冷冻瓶长期使用）。3. 预分离生产了7万个馏分，到2022年底，生产的总馏分将达到58万个。4. NPNPD镀了500万口馏分井，供应给校外社区。5. NPNPD目前有10个已执行的合作协议，2个crada， 5个cda和另外19个mta来接收预分馏样品。6. NPNPD在2022年又收到了7份mta接收NPNPD分数的请求。7. 与NIAID启动了一项新的机构间协议，并继续与NCCIH和NCATS进行机构间合作。与癌症预防部发起了一个新的联合项目。还与伊利诺伊大学芝加哥分校启动了新的CRADA，并与诺华公司达成了合作协议。NPNPD还向研究实验室运送了额外的1,900,000个预分馏样品进行筛选。9. NPNPD化学实验室对预分离样品筛选中鉴定的生物活性组分进行了1386次二次纯化，并将30,000份高度纯化的样品送回筛选中心。2. 解决倡议和癌症登月计划的目标a.描述为实现倡议的目标和结果所取得的主要成就。生产58万份天然产品预分馏样品。已准备好384孔板，其中包含20,000,000个预分馏样品，准备发货。向公众发布了第一批50万个分数。在执行50个mta后，向校外筛选实验室运送了7,200,000个电镀筛选样品。从NPNPD馏分库的筛选中鉴定出8万个纯化的活性馏分。创建了新的微生物库，其中包括24,000种美国土壤真菌和6500种澳大利亚海洋微生物。将技术转让给多个校外研究中心（南非、巴西、韩国、瑞典）。发现了一种具有潜在抗癌活性的新型激酶抑制剂，并已获得NCI专利。鉴定出一种新的环状肽，这是TDP1酶的第一个变构抑制剂，已获得NCI专利，并与Genentech达成合作协议。在弗吉尼亚理工学院和州立大学、哈佛医学院、澳门大学、圣保罗大学（巴西）、NIAID药物开发兴趣小组、北卡罗来纳大学格林斯博罗分校、国家补充和综合健康中心、比勒陀利亚大学、西开普省大学、开普敦大学和罗德岛大学（南非）以及国家癌症咨询委员会发表了关于NPNPD的特邀演讲。发表论文4篇，其中3篇登上各自期刊封面[Evans JR， Akee RK, Chanana S, McConachie GD, Thornburg CC, Grkovic T， O'Keefe BR]。国家癌症研究所（NCI）天然产物发现计划：利用人工神经网络探索NCI-60天然产物样本筛选数据。ACS Omega. 8(10):9250- 9256,2023；Wilson BAP, Li N, Martinez Fiesco JA, Dalilian M, Wang D, Smith EA, Wamiru A, Shah R, Goncharova EI, Beutler JA, Grkovic T，张平，O'Keefe BR。合成和天然产物腺苷3′，5′-环单磷酸依赖蛋白激酶A （PKA）抑制剂的生化发现、细胞内评价和晶体学表征。中国药理学杂志，6(4):633-650,2023；孔涛，李德，Wilson BAP, Ranguelova K, Dalilian M, Smith EA, Wamiru A, Goncharova EI, Grkovic T, Voeller D, Lipkowitz S, Schnermann MJ， O'Keefe BR, Du L. Plakinidine B光化学二聚化对E3泛素蛋白连接酶ccl -B的抑制作用。组织通讯，24:9468-9472,2022.；Martinez-Fructuoso L;Arends, s.j.r.；弗莱雷，V. F.；埃文斯，j.r.；DeVries,美国;Peyser, b.d.；阿基，r.k.；索恩伯格，c.c.；库马尔,r;Ensel,美国;摩根，通用汽车；麦康纳博士；Veeder:;邓肯，l.r.；Grkovic t;O'Keefe， B. R.从NCI天然产物发现预分离提取物库中筛选新的抗菌天然产物。中华医学会传染病学杂志，2016 [j]。b.关键成就如何实现推荐目标？关键成就表明，NPNPD已经创建并减少了自动化系统的实践，这些系统大大提高了支持药物发现的天然产物化学工作的速度和功能。它们还显示了校外研究团体的广泛“支持”，以及其他美国政府机构对NPNPD影响的扩展。NPNPD出版物在出版时受到高度追捧和特色，增加了该计划的知名度，并且每年都会收到关于NPNPD研讨会的多个请求。NPNPD正在实现其目标，即在药物发现项目中增加天然产物样本的使用，目前已完成多个筛选，已确定了生物活性天然产物。c.加强合作是癌症登月计划的主要目标。请描述该计划的协作活动（如鼓励协作的补充，网络之间的协作研究等）。NPNOD是高度协作的。这些都是NCI天然产物化学和校外筛选中心之间的合作。我们还完成了与几个美国政府机构的合作协议。我们越来越多地看到NPNPD组分筛选产生新的生物活性分子的结果，包括最近被NCI申请专利的一项发现。最后，我们继续与阿斯利康进行CRADA，以筛选针对肿瘤靶点的NPNPD组分，并与诺华完成了CDA。d.保健差距和各组织之间的伙伴关系等跨领域主题是癌症登月计划的主要组成部分。请描述该倡议是如何处理跨领域主题的？NPNPD已经在跨领域的NIH小组中进行合作，包括NIH HEAL倡议与NCATS、NCCIH和校外研究小组。我们正在与NIAID合作开发针对ESKAPE病原体的抗生素，并与WRAIR合作为国防部开发抗疟疾和抗利什曼病药物。美国国家海洋和大气管理局（NOAA）已经启动了另一项海洋收集RFA， NCCIH为全球天然产物核磁共振数据库发布了一项拨款。我们还与NCI癌症预防部启动了一个联合项目，该部门将资助使用NPNPD分数的筛查项目。

1. Overall progress a. List the expected goals and outcomes of the initiative. Aim 1. Create new technologies to build an enhanced NP pre-fractionated library amenable to modern high-throughput targeted screening programs. Aim 2. Expand the chemical diversity available to the public from culturable microorganisms with new methods and libraries. Aim 3. Provide the pre-fractionated library to screening centers worldwide to accelerate drug discovery. Aim 4. Provide high throughput screening support for researchers to enable targeted discovery efforts. Aim 5. Provide faster analytical resources (isolation, structure elucidation, re-supply) to expedite translational pipelines. Aim 6. Establish a public database and bioinformatics platform to broaden input and expand impact. b. Summarize progress that has been made in the reporting period (FY22). 1. In FY 2022 the NPNPD reached a level of 18,000 soil fungi which were plated, photographed, and cryo-vialed for long term use. In addition, 410 large-scale fermentations and extractions were conducted. 2. The genetic sequencing and taxonomic determination of 1000 marine actinomycetes and 1000 marine fungi (which are also being plated, photographed, and cryo-vialed for long term use) was also completed. 3. Prefractionation efforts produced 70,000 fractions to bring the total factions produced to 580,000 by the end of 2022. 4. The NPNPD plated 5,000,000 wells of fractions to supply to the extramural community. 5. The NPNPD currently has 10 executed collaboration agreements, 2 CRADAs, 5 CDAs, and an additional 19 MTAs to receive pre-fractionated samples. 6. The NPNPD received an additional 7 requests for MTAs to receive NPNPD fractions in 2022. 7. Initiated a new interagency agreement with NIAID and continued IAAs with NCCIH and NCATS. Initiated a new joint program with the Division of Cancer Prevention. Also initiated new CRADA with the University of Illinois at Chicago and a Collaboration Agreement with Novartis. 8. The NPNPD shipped an additional 1,900,000 pre-fractionated samples to research labs for screening. 9. The NPNPD Chemistry laboratory undertook 1,386 secondary purifications of identified bioactive fractions identified in screens of pre-fractionated samples and returned 30,000 highly purified samples back to screening centers. 2. Addressing initiative and Cancer Moonshot goals a. Describe key accomplishments that have been made towards achieving the Initiative's goals and outcomes. Production of 580,000 pre-fractionated natural product samples. Have prepared 384-well plates containing 20,000,000 pre-fractionated samples ready for shipping. Released first 500,000 fractions to the public. Shipped 7,200,000 plated screening samples to extramural screening laboratories after execution of 50 MTAs. Produced 80,000 purified sub-fractions of active fractions identified in screens of the NPNPD fraction library. Created new microbial library with 24,000 U.S. soil fungi and 6500 marine microbes from Australia. Transferred technologies to multiple extramural research centers (S. Africa, Brazil, Korea, Sweden). Identified a novel kinase inhibitor with potential anticancer activity that has been patented by the NCI. Identified a novel cyclic peptide that is the first allosteric inhibitor of the enzyme TDP1 which haas been patented by the NCI and is the subject of a collaboration agreement with Genentech. Presented invited talks on the NPNPD to the Virginia Polytechnic Institute and State University, Harvard Medical School, University of Macau, University of Sao Paulo (Brazil), NIAID Drug Development Interest Group, University of North Carolina at Greensboro, National Center for Complementary and Integrative Health, Universities of Pretoria, Western Cape, Cape Town and Rhodes (S. Africa), and the National Cancer Advisory Board. Published 4 manuscripts 3 of which featured on the cover of their respective journals [Evans JR, Akee RK, Chanana S, McConachie GD, Thornburg CC, Grkovic T, O'Keefe BR. National Cancer Institute (NCI) Program for Natural Product Discovery: Exploring NCI-60 Screening Data of Natural Product Samples with Artificial Neural Networks. ACS Omega. 8(10):9250-9256, 2023; Wilson BAP, Li N, Martinez Fiesco JA, Dalilian M, Wang D, Smith EA, Wamiru A, Shah R, Goncharova EI, Beutler JA, Grkovic T, Zhang P, O'Keefe BR. Biochemical Discovery, Intracellular Evaluation, and Crystallographic Characterization of Synthetic and Natural Product Adenosine 3',5'-Cyclic Monophosphate-Dependent Protein Kinase A (PKA) Inhibitors. ACS Pharmacol Transl Sci. 6(4):633-650, 2023.; Khong QT, Li D, Wilson BAP, Ranguelova K, Dalilian M, Smith EA, Wamiru A, Goncharova EI, Grkovic T, Voeller D, Lipkowitz S, Schnermann MJ, O'Keefe BR, Du L. Photochemical Dimerization of Plakinidine B Leads to Potent Inhibition of the E3 Ubiquitin-Protein Ligase CBL-B. Org Lett. 24:9468-9472, 2022.; Martinez-Fructuoso, L; Arends, S. J. R.; Freire, V. F.; Evans, J. R.; DeVries, S.; Peyser, B. D.; Akee, R. K.; Thornburg, C. C.; Kumar, R.; Ensel, S.; Morgan, G. M.; McConachie, G. D.; Veeder, N.; Duncan, L. R.; Grkovic, T.; and O'Keefe, B. R. A screen for new antimicrobial natural products from the NCI Program for Natural Product Discovery prefractionated extract library. ACS Infectious Diseases, 2023]. b. How do key accomplishments address recommendation goals? The key accomplishments show that the NPNPD has created and reduced to practice the automated systems necessary to greatly increase the speed and functionality of natural products chemistry efforts in support of drug discovery. They also show broad "buy-in" by the extramural research community as well as extension of NPNPD impact by other U.S. Government agencies. NPNPD publications are highly sought and featured when published, increasing the visibility of the program and multiple requests for seminars on the NPNPD are received on a yearly basis. The NPNPD is achieving its goal of increasing the use of natural product samples in drug discovery programs with multiple screens now completed that have identified bioactive natural products. c. Enhancing collaboration is a major goal of the Cancer Moonshot. Please describe the initiative's collaboration activities (such as supplements to encourage collaboration, collaborative research between networks, etc.). The NPNOD is highly collaborative. Each of these is a collaboration between NCI natural products chemistry and extramural screening centers. We have also completed collaboration agreements with several U.S. Government agencies. We are increasingly seeing the results of screens with NPNPD fractions resulting in new bioactive molecules including a recent discovery being patented by the NCI. Finally, we continued a CRADA with Astra Zeneca to screen NPNPD fractions against an oncology target and have completed a CDA with Novartis. d. Cross-cutting themes of health disparities and partnerships between organizations are major components of the Cancer Moonshot. Please describe how cross-cutting themes are being addressed within the initiative? The NPNPD is already collaborating in cross-cutting NIH groups including the NIH HEAL initiative with NCATS, NCCIH and extramural research groups. We are working with NIAID on antibiotic discovery against ESKAPE pathogens and with WRAIR on anti-malarial and anti-leishmaniasis agent discovery for the DOD. NOAA has initiated another RFA for the collection of marine collections and NCCIH issued a grant for a global natural product NMR database. We also have initiated a joint program with the NCI Division of Cancer Prevention who will be funding screening projects using NPNPD fractions.