Tetrahydroisoquinoline Antitumor Drugs: Synthesis and Biosynthesis

四氢异喹啉抗肿瘤药物：合成与生物合成

• 批准号: 8193086
• 负责人: Robert Michael Williams
• 金额: $ 30.87万
• 依托单位: COLORADO STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-04-10 至 2013-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8193086
• 关键词: Affinity; Affinity Labels; Alkaloids; Amino Acid Sequence; Anabolism; Antineoplastic Agents; Binding; Biochemical; Bioinformatics; Biological; Biological Factors; Biotin; Carbohydrates; Carrier Proteins; Cell Separation; Cells; Chemicals; Chemistry; Clinical; Complex; DNA; DNA Alkylation; DNA Sequence; DNA crosslink; DNA-Binding Proteins; Databases; Development; Early identification; Ecteinascidin 743; Engineering; Enzymes; Evaluation; Family; Florida; Fourier transform ion cyclotron resonance; Gene Cluster; Genetic Engineering; Goals; Grant; Health; Investigation; Label; Laboratories; Ligation; Link; Marines; Metagenomics; Methodology; Methods; Molecular; Molecular Structure; Nature; Nucleic Acids; Oxidation-Reduction; Pathway interactions; Peptide Sequence Determination; Pharmaceutical Preparations; Physical condensation; Population; Production; Proteins; Sequence Analysis; Series; Staging; Streptavidin; Sulfides; Superoxides; Synthesis Chemistry; System; Technology; Tetrahydroisoquinolines; Transferase; Tyrosine; Urochordata; adduct; affinity labeling; analog; antineoplastic antibiotics; antitumor drug; base; chemical addition; chemical synthesis; clinically relevant; crosslink; cyanonaphthyridinomycin; design; drug synthesis; genome sequencing; human ZNF9 protein; lemonomycin; member; metagenomic sequencing; microbial; microbial community; microbial host; microorganism; new technology; oxidation; oxidative damage; peptide synthase; positional cloning; programs; rRNA Genes; tetrazomine; tool

DESCRIPTION (provided by applicant): The primary focus of this application is to study the total synthesis and biosynthesis of complex, biomedically significant natural products constituted of tetrahydroisoquinolines. The synthetic chemistry that will be developed shall be utilized to prepare analogues of the natural substances as biological, biosynthetic and mechanistic probes. This proposal is primarily hypothesis-driven, and extensively employs new synthetic methodologies developed in this laboratory for the construction of such agents. The specific aims of this program are to study the interaction of the natural antitumor antibiotics and mechanistically inspired synthetic analogs, including ecteinascidin 743 (Et-743), saframycin, jorumycin, tetrazomine, lemonomycin and the bioxalomycins with cellular nucleic acids. The DNA-alkylating capacity of these drugs compared with their ability to cause oxidative damage to nucleic acids will be explored. The synthesis of members of this class of antitumor drugs will continue to be developed with the objective of harnessing the synthetic methodology developed to make new, less toxic, more selective and more potent antitumor drugs. In addition, the tools of synthesis will be exploited to synthesize mechanistic probes for the interaction of these substances with cellular nucleic acids and proteins that bind to cellular nucleic acids. We have recently discovered that bioxalomycin 12 specifically cross-links duplex DNA at 54dCpG34 steps. We propose to elucidate the exact molecular structure of the covalent adduct. This finding has inspired new design concepts for simpler analogs based on the tetrahydroisoquinoline core that may be capable of alkylating and cross-linking DNA as well as potentially cross-linking DNA to DNA-binding proteins. The antiproliferative activity of this family of alkaloids is intimately associated with the capacity of these agents to bind to and covalently modify DNA. This is particularly evident in the case of Et-743. Plans are presented to mechanistically separate the DNA alkylation and cross-linking chemistry of these agents from their capacity to inflict oxidative damage on cells. The investigation of the biosynthesis of Et-743 with a particular focus on the late-stage assembly of the macrocyclic sulfide-bridged spiro-tetrahydroisoquinoline ring system will be pursued. We plan to apply high throughput genome sequencing methods using 454 technology to identify the Et-743 biosynthetic gene cluster from Ecteinascidia turbinata. The ultimate goals of the biosynthesis studies are to genetically engineer a fermentable microorganism to produce Et-743 for clinical use. PUBLIC HEALTH RELEVANCE: The purpose of this application is to utilize the tools of chemical synthesis to study the molecular details of how Nature biosynthesizes anti-cancer drugs. In particular, this program will deploy new technologies to produce the clinically relevant anticancer drug ecteinascidin 743 (Et-743) on industrially practical scale through genetic engineering of a microbial host. In addition, the chemical technologies being developed will be applied to the design and synthesis of new anti-cancer drugs.

描述（由申请人提供）：本申请的主要重点是研究由四氢异喹啉组成的复杂的、具有生物医学意义的天然产物的全合成和生物合成。将开发的合成化学应用于制备天然物质的类似物，作为生物、生物合成和机械探针。该提案主要是假设驱动的，并广泛采用本实验室开发的新的合成方法来构建此类试剂。该计划的具体目的是研究天然抗肿瘤抗生素和机械启发的合成类似物，包括海鞘素743（Et-743），沙夫霉素，jorumycin，tetrazomine，lemonomycin和biobacteromycins与细胞核酸的相互作用。将探讨这些药物的DNA烷基化能力与其对核酸造成氧化损伤的能力。这类抗肿瘤药物的合成将继续发展，目的是利用已开发的合成方法来制造新的、毒性更低、选择性更高和效力更强的抗肿瘤药物。此外，将利用合成工具来合成这些物质与细胞核酸和与细胞核酸结合的蛋白质相互作用的机械探针。我们最近发现，双曲霉素12在54 dCpG 34步骤特异性交联双链体DNA。我们建议阐明共价加合物的确切分子结构。这一发现激发了基于四氢异喹啉核心的更简单类似物的新设计概念，这些类似物可能能够烷基化和交联DNA以及潜在地将DNA交联到DNA结合蛋白。该生物碱家族的抗增殖活性与这些药剂结合并共价修饰DNA的能力密切相关。这在Et-743的情况下尤其明显。计划提出了机械分离的DNA烷基化和交联化学的这些代理商从他们的能力造成氧化损伤细胞。Et-743的生物合成的调查，特别侧重于后期组装的大环硫桥螺-四氢异喹啉环系统将继续进行。我们计划应用454技术的高通量基因组测序方法来鉴定Et-743生物合成基因簇。生物合成研究的最终目标是对可发酵的微生物进行基因工程改造，以生产临床使用的Et-743。公共卫生相关性：该应用程序的目的是利用化学合成的工具来研究大自然如何生物合成抗癌药物的分子细节。特别是，该计划将部署新技术，通过微生物宿主的基因工程，以工业实用规模生产临床相关的抗癌药物ecteinascidin 743（Et-743）。此外，正在开发的化学技术将用于设计和合成新的抗癌药物。