Biosynthesis of cytotoxic sesquiterpenoids by higher basidiomycetes

高等担子菌生物合成细胞毒性倍半萜类化合物

• 批准号: 7368651
• 负责人: CLAUDIA SCHMIDT-DANNERT
• 金额: $ 28.93万
• 依托单位: UNIVERSITY OF MINNESOTA
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-09-22 至 2012-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7368651
• 关键词: Agaricales; Anabolism; Anti-Bacterial Agents; Antibiotics; Attention; Basidiomycota; Biological Factors; Class; Complementary DNA; Coprinus; Cosmids; Cyclization; Cyclobutanes; Cyclopropanes; Cytochrome P450; Cytostatics; Data; Enzymes; Exhibits; Fruit; Genes; Goals; Human; Investigation; Label; Libraries; Literature; Metabolic Pathway; Microbe; Mixed Function Oxygenases; Modification; Multi-Drug Resistance; Oligonucleotide Primers; Oligonucleotide Probes; Pharmaceutical Preparations; Phase I/II Trial; Plants; Polymerase Chain Reaction; Property; Range; Reaction; Recombinants; Reporting; Research; Resources; Saccharomyces cerevisiae; Screening procedure; Sesquiterpenes; Skeleton; Solid Neoplasm; Structure; Therapeutic; Therapeutic Index; Thinking; Traditional Medicine; Tumor Cell Line; acylfulvene; analog; antitumor drug; base; cyclopropane; cytotoxic; farnesyl pyrophosphate; feeding; fungus; illudin M; improved; isoprenoid; neoplastic cell; novel; oxidation; scaffold; substance R; tumor

DESCRIPTION (provided by applicant): Higher basidiomycetes, commonly referred to as mushrooms, have been used for centuries in traditional medicine and are known to synthesize an array of biologically active natural products. It is thought that mushrooms are among the greatest unexplored resources for the discovery of novel biologically active compounds. Despite the extremely rich chemodiversity of higher basidiomycetes and the prospects for the discovery of new drugs, mushrooms have only recently caught the attention of medicinal chemists and biochemists. Mushrooms are known to produce a variety of sesquiterpenes with unique skeletons not found in microbes or plants. One class of sesquiterpenoids, the illudanes, has received considerable attention because of their potent antitumor and antibacterial activity. Illudins M and S produced by the Jack O'Lantern mushroom Omphalotus olearius exhibit cytotoxic and cytostatic properties at nanomolar concentrations in several human tumor cell lines, including multiple drug resistant tumors. Semi-synthetic illudin analogues irofulvene and acylfulvene with improved therapeutic indexes and remarkable antitumor activities have been synthesized and are currently under investigation in several phase I and II trials against several solid tumor types. The biosynthesis, however, of this class of illudane sesquiterpenoids is not known. The proposed research will be the first biosynthetic investigation of this important and unique class higher fungal sesquiterpene natural products. The specific hypotheses of this proposal are that: (1) Illudins are derived via a protoilludane skeleton synthesized by a new type of cyclase and (2) installment of the reactive cyclopropane ring in illudins occurs after the initial cyclization reaction presumably via a P450 monooxygenase catalyzed reaction. It is proposed to (1) identify and characterize the cyclase responsible for formation of the protoilludane scaffold, (2) identify and characterize additional illudin biosynthetic genes and (3) probe the substrate range of protoilludane scaffold modifying enzymes. The long-term goal of this research is to investigate biosynthesis of different protoilludane-type sesquiterpenoids in higher fungi with the goal of using the identified biosynthetic enzymes to explore synthesis of new functionalized sesquiterpenoids with potent antitumor properties and improved therapeutic properties. It is thought that mushrooms are among the greatest unexplored resources for the discovery of novel biologically active compounds; but despite the extremely rich chemodiversity of higher basidiomycetes and the prospects for the discovery of new drugs, mushrooms have only recently caught the attention of medicinal chemists and biochemists. One class of sesquiterpenoids, the illudanes, produced by the Jack O'Lantern mushroom Omphalotus olearius exhibit potent cytotoxic and cytostatic properties in several human tumor cell lines, including multiple drug resistant tumors. The goal of this research is to investigate biosynthesis of illudane sesquiterpenoids in mushrooms with the goal of using the identified biosynthetic enzymes to explore synthesis of new sesquiterpenoid structures with potent antitumor properties and improved therapeutic properties.

描述（由申请人提供）：高等担子菌，通常称为蘑菇，已经在传统医学中使用了几个世纪，并且已知可以合成一系列具有生物活性的天然产物。人们认为蘑菇是发现新型生物活性化合物的最大的未开发资源之一。尽管高等担子菌具有极其丰富的化学多样性以及发现新药的前景，但蘑菇直到最近才引起药物化学家和生物化学家的注意。 众所周知，蘑菇可以产生多种倍半萜，这些倍半萜具有微生物或植物中未发现的独特骨架。一类倍半萜类化合物伊鲁丹类化合物因其有效的抗肿瘤和抗菌活性而受到相当大的关注。杰克灯笼蘑菇 Omphalotus olearius 产生的 Illudins M 和 S 在纳摩尔浓度下在多种人类肿瘤细胞系（包括多种耐药肿瘤）中表现出细胞毒性和细胞抑制特性。具有改善的治疗指数和显着抗肿瘤活性的半合成伊洛伞素类似物irofulvene和acylfulvene已经被合成，目前正在针对几种实体瘤类型的几项I期和II期试验中进行研究。 然而，这类伊鲁丹倍半萜类化合物的生物合成尚不清楚。拟议的研究将是对这种重要且独特的高级真菌倍半萜天然产物的首次生物合成研究。该提案的具体假设是：（1）隐杯伞素是通过新型环化酶合成的原伊伞烷骨架衍生的；（2）隐杯伞素中反应性环丙烷环的安装发生在可能是通过 P450 单加氧酶催化的初始环化反应之后。反应。建议(1)鉴定和表征负责形成原伊鲁丹支架的环化酶，(2)鉴定和表征额外的乌鲁丹生物合成基因，以及(3)探测原伊鲁丹支架修饰酶的底物范围。 本研究的长期目标是研究高等真菌中不同原伊鲁丹型倍半萜类化合物的生物合成，目的是利用已鉴定的生物合成酶探索合成具有有效抗肿瘤特性和改善治疗特性的新型功能化倍半萜类化合物。人们认为蘑菇是发现新型生物活性化合物的最大的未开发资源之一。尽管高等担子菌具有极其丰富的化学多样性，并且具有发现新药的前景，但蘑菇直到最近才引起药物化学家和生物化学家的注意。由杰克灯笼蘑菇 Omphalotus olearius 产生的一类倍半萜类化合物伊鲁丹 (illudanes) 在多种人类肿瘤细胞系（包括多种耐药肿瘤）中表现出有效的细胞毒性和细胞抑制特性。本研究的目的是研究蘑菇中伊鲁丹倍半萜类化合物的生物合成，目的是利用已鉴定的生物合成酶来探索具有有效抗肿瘤特性和改善治疗特性的新倍半萜类结构的合成。