Enediyne Biosynthesis and Engineering

烯二炔生物合成与工程

• 批准号: 7811497
• 负责人: Ben Shen
• 金额: $ 17.84万
• 依托单位: UNIVERSITY OF WISCONSIN-MADISON
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-09-30 至 2011-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7811497
• 关键词: Active Sites; Acyl Carrier Protein; American; Anabolism; Antineoplastic Agents; Biochemical; Biological; Biological Factors; C 1027; Cancer Etiology; Cells; Cessation of life; Characteristics; Chemicals; Chemistry; Clinical; Collaborations; Complex; Coupling; Development; Dynemicin; Engineering; Enzymatic Biochemistry; Enzymes; Family; Funding; Genes; Genetic; Genome; Genome Scan; Goals; Grant; Health; Hypoxia; In Vitro; Malignant Neoplasms; Maps; Medical; Methodology; Methods; Mining; Natural Products Chemistry; Outcome; Outcome Study; Parents; Pathway interactions; Peptides; Peripheral; Pharmaceutical Preparations; Polyenes; Polymers; Production; Property; Reaction; Recombinants; Recovery; Research; Roentgen Rays; Role; Scanning; Source; Streptomyces; Structure; System; Testing; United States National Institutes of Health; X-Ray Crystallography; Zinostatin; analog; antibody conjugate; antitumor agent; base; cancer cell; chemotherapy; chromophore; clinical application; combinatorial; cytotoxicity; drug discovery; genetic manipulation; in vivo; microbial; microbial genome; microorganism; novel; parent grant; polyketide synthase; programs; public health relevance; response; structural genomics; success

DESCRIPTION (provided by applicant): In response to NIH NOT-OD-09-058 titled "NIH Announces the Availability of Recovery Act Funds for Competitive Revision Applications," we wish to extend studies spearheaded during the funding of NIH grant 2R01 CA78747 titled "Enediyne Biosynthesis and Engineering." Cancer causes one of every four deaths in the US. The development of fundamentally new, clinically useful anticancer drugs therefore constitutes a national health and research imperative. The enediynes are the most potent, highly active anticancer agents in existence today, and their use as anticancer drugs has been demonstrated clinically. A great challenge is to develop ways to prepare enediynes and their structural analogs and to discover new enediyne natural products for mechanistic studies and clinical development. We propose in this Competitive Revision application to (1) structurally (by X-ray crystallographic means) characterize the enediyne polyketide synthases (PKSEs) and their associated enzymes for enediyne core biosynthesis as well as other enzymes from selected 9- and 10- membered enediyne biosynthetic pathways; (2) produce and further analyze engineered enediynes with distinct exploitable biophysical properties lending themselves to potential clinical applications; and (3) isolate and characterize new enediynes from microbial sources identified on the basis of genome mining. Our hypotheses are that: (1) characterization of selected novel enzymes involved in enediyne biosynthesis especially C-1027, neocarzinostatin (NCS), maduropeptin (MDP), calicheamicin (CAL), esperamicin (ESP), and dynemicin (DYN)) biosynthesis will make fundamental contributions to mechanistic enzymology and natural product chemistry; (2) enediynes produced by combinatorial biosynthetic methods can and do display biological activities superior to those displayed by the parent compound; such compounds warrant further study enabled only through increased production; and (3) new microorganisms identified on the basis of genome mining produce novel, and potentially medically important, enediynes. The specific aims for this Competitive Revision application are: (1) In vivo and in vitro characterization of the selected enediyne PKSs and associated enzymes and their roles in both 9- (C-1027, NCS, and MDP) and 10-membered (CAL, ESP, and DYN) enediyne core biosynthesis; (2) Structural characterization of selected enzymes from enediyne (C- 1027, NCS, MDP, CAL, and DYN) biosynthetic machineries by X-ray crystallography; (3) Isolation of engineered C-1027 analogs to evaluate them as anticancer agents in vivo; and (4) Isolation and structural elucidation of novel 9- or 10-membered enediyne natural products from S. ghanaensis NRRL B-12104, A. orientalis ATCC43491, and S. citricolor IFO13005. The outcomes from these studies will greatly accelerate the tempo of our enediyne biosynthesis, engineering, and drug discovery program by (1) defining the minimal enzymes necessary to convert a nascent linear polyene intermediate from the enediyne PKSE to the characteristic enediyne core structure, (2) demonstrating, on a pilot scale, the feasibility of a "structural genomics" approach to enediyne biosynthesis by solving the structures of key enzymes from selected pathways, (3) advancing C-1027 and its engineered analogs into in vivo testing to realistically develop them into clinically useful, new anticancer drugs, and (4) expanding the portfolio of enediyne anticancer drugs and drug leads by isolating new enediyne natural products. PUBLIC HEALTH RELEVANCE: Cancer causes 1 of every 4 deaths in the US, and 565,650 Americans are expected to die of cancer in 2008. It is therefore a critical research goal to optimize available drugs and to develop fundamentally new, clinically useful anticancer drugs. The enediynes are the most potent, highly active anticancer agents in existence today. Although the natural enediynes have seen limited use as clinical drugs, polymer-based delivery systems and enediyne-antibody conjugates have shown great clinical success or promise in anticancer chemotherapy, demonstrating that the enediynes can be developed into powerful drugs when their extremely potent cytotoxicity is harnessed and delivered to specific cancer cells. A great challenge is to develop methods to make enediynes and their structural analogs and to discover new enediyne natural products for mechanistic studies and clinical developments. This research will study enediyne biosynthesis and engineered novel enediyne analogs. The outcomes include development of enediynes and their analogs into potential anticancer drugs.

描述（由申请人提供）：响应NIH NOT-OD-09-058，标题为“ NIH宣布恢复ACT的可用性用于竞争性修订应用程序”，我们希望扩展在NIH授予NIH 2R01 CA78747的资金期间先生的研究，名为“ Enediyne Biosynsis和Engineering”。癌症在美国每四个死亡之一。因此，从根本上新的，临床上有用的抗癌药的发展构成了国家健康和研究。 ENEDIYNES是当今现有的最有效，最活跃的抗癌药，它们用作抗癌药物的使用已在临床上证明。一个巨大的挑战是开发制备EneDiynes及其结构类似物的方法，并发现新的Enediyne天然产品用于机械研究和临床开发。我们在此竞争性修订应用中提出了（1）结构上（X射线晶体学均值），该晶体（通过X射线晶体学方式）表征了Enediyne Polyketide合酶（PKSE）及其相关酶，用于Enediyne核心生物合成以及其他来自所选的9-和10成员的Enediyne生物合成途径的酶； （2）生产并进一步分析了具有独特的可利用生物物理特性的工程化含量，这些特性借给潜在的临床应用； （3）孤立并表征了根据基因组挖掘确定的微生物来源的新endiynes。我们的假设是：（1）表征参与Enediyne生物合成的选定新型酶，尤其是C-1027，Neocarzinototatin（NCS），Maduropeptin（MDP）（MDP），Calicheamicin（Cal），Esperamicin（ESP）（ESP）和贡献效果，使生物合作构成了机构的贡献，以使其及其贡献进行了贡献。 （2）组合生物合成方法产生的endiynes可以并且确实显示出比父院所表现出的生物学活性；这样的化合物只有通过增加产量才能进一步进行研究。 （3）根据基因组采矿鉴定的新微生物产生了新颖的，并且潜在的医学重要性含量。此竞争性修订应用的具体目的是：（1）所选endiyne PKS和相关酶的体内和体外表征及其在9-（C-1027，NCS和MDP）和10-Merded（Cal，ESP，ESP和DYNENENE CORE CORE BIOSYNTHESOS）中的作用； （2）通过X射线晶体学从Enediyne（C- 1027，NCS，MDP，CAL和DYN）生物合成机器中选定的酶的结构表征； （3）分离工程的C-1027类似物，以评估它们为体内的抗癌药； （4）从S. ghanaensis nrrl B-12104，A。Orientalis atcc43491和S. citricolor ifo13005中，新的9或10元的Enediyne天然产物的新型9或10元的Enediyne天然产物的分离和结构阐明（4）。 The outcomes from these studies will greatly accelerate the tempo of our enediyne biosynthesis, engineering, and drug discovery program by (1) defining the minimal enzymes necessary to convert a nascent linear polyene intermediate from the enediyne PKSE to the characteristic enediyne core structure, (2) demonstrating, on a pilot scale, the feasibility of a "structural genomics" approach to通过从选定途径中求解关键酶的结构，（3）将C-1027及其工程类似物推向体内测试，从而将它们分成临床上有用的新抗癌药物，以及（4）通过扩大新的抗癌药物的投资组合，将它们隔离开发。 公共卫生相关性：癌症导致美国每4例死亡中的1例，预计565,650名美国人将在2008年死于癌症。因此，优化可用药物并开发出根本新的，临床上有用的抗癌药物是一个关键的研究目标。 ENEDIYNES是当今现有的最有效，最活跃的抗癌药。尽管天然endiynes作为临床药物的使用有限，但聚合物基于聚合物的递送系统和ENEDIYNE-ANTIPODY结合物在抗癌化疗中表现出巨大的临床成功或有希望，表明当它们可以将其非常有效的细胞毒性置于轴承并递送到特定的癌症细胞中时，可以将其开发为强大的药物。一个巨大的挑战是开发方法来制作EneDiynes及其结构类似物，并发现用于机械研究和临床发展的新Enediyne天然产品。这项研究将研究Enediyne生物合成和设计的新型Enediyne类似物。结果包括将Enediynes及其类似物的开发为潜在的抗癌药物。