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Combinatorial biosynthesis and metabolism studies of novel tetracenomycins

新型四烯霉素的组合生物合成和代谢研究

基本信息

批准号：
10045418
负责人：
Eric Nybo
金额：
$ 38.54万
依托单位：
FERRIS STATE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-08-01 至 2023-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10045418
关键词：
Address Anabolism Anthracycline Antibiotics Antineoplastic Agents Area Automobile Driving Binding Biogenesis Biological Assay Cancer cell line Chemicals Clinic Cytochromes DNA Data Deoxy Sugars Development Dose-Limiting Doxorubicin Drug usage Enzymes Exhibits Genes Genetic Engineering Genomics Glycosyltransferase Gene Goals Hepatic High Pressure Liquid Chromatography Human Institution Knowledge Literature Liver Malignant Neoplasms Metabolic Metabolism Michigan Microsomes Minor Groove Modification NMR Spectroscopy Natural Products Pathway interactions Pattern Penetration Pharmaceutical Preparations Phase Plasmids Preparation Production Property Public Health Publishing Reporting Research Research Infrastructure Rhamnose Role Solid Neoplasm Streptomyces Streptomyces lividans Structure Structure-Activity Relationship System Testing Therapeutic Topoisomerase II Universities Work analog anticancer activity base cancer cell cancer therapy combinatorial drug development drug discovery drug metabolism evidence base flexibility glycosylation glycosyltransferase improved in vivo innovation insight intercalation leukemia liver metabolism metabolic abnormality assessment metabolic profile novel novel therapeutics sugar treatment research undergraduate student

项目摘要

Project Summary/Abstract Combinatorial biosynthesis and metabolism studies of novel tetracenomycins Anthracyclines are natural product antibiotics that are among the most effective anticancer drugs used in the clinic. Biosynthetic modification of anthracyclines is a promising strategy to generate new chemical analogs with differentiated anticancer activities and expanded therapeutic windows. Elloramycin (ELM, 1) is an anthracycline antibiotic produced by Streptomyces olivaceus Tü 2353 that features a tetracyclic elloramycinone aglycone and an appended 8-O-2,3,4-tri-O-methyl-a-L-rhamnose sugar. Elloramycins and tetracenomycins (TCMs) exhibit mild antiproliferative activity due to inhibition of DNA topoisomerase II. Furthermore, optimization of the deoxysugar moiety is essential for refining the cellular penetration, potency, clearance, and metabolism of the analogs. However, we still do not fully understand the structure-activity-relationships of the TCM deoxysugar on antiproliferative activity and drug metabolism. Furthermore, despite preparation of >20 different analogs of ELM, the SAR of these analogs has not yet been assessed rigorously in a panel of cancer cell lines. Our long-term goal is to produce anthracycline analogs with improved antiproliferative activity. To facilitate production of analogs, we will exploit the S. lividans (cos16F4) expression system to alter the deoxysugar moiety of tetracenomycins via combinatorial biosynthesis. This comprehensive platform will facilitate development of new drug leads for use in human cancer. The overall objectives of the proposed research are three-fold: (1) to synthesize novel TCM derivatives, (2) to validate the anticancer activity of the TCMs, and (3) to develop a S9 fraction assay to evaluate metabolism of 1 and the most active analogs. Our hypotheses are that (1) ElmGT will be “substrate-flexible” enough to transfer the intended TDP-deoxysugar donors to 8-demethyl-tetracenomycin C, (2) the appended deoxysugar will alter the anticancer activity via binding to topoisomerase II and DNA, and (3) it will alter drug metabolism via differential binding to hepatic cytochrome P450s. Our specific aims will test these hypotheses: (Aim 1) we will heterologously express “sugar plasmids” in S. lividans (cos16F4) to alter the TCM deoxysugar moiety, (Aim 2) we will evaluate the antiproliferative activity of the new TCM analogs, (Aim 3) we will develop a S9 fraction assay to determine the metabolic profile of TCM analogs. The rationale is that the “substrate-flexible” glycosyltransferase, ElmGT, should accept the novel TDP-deoxysugar donors. This contribution is significant because the expected production platform will more fully explore the chemical space of TCM analogs. Furthermore, this research is innovative because we are investigating the impact of deoxysugar modifications on antitumoral activity and metabolism of tetracenomycins, drug properties which are currently not well-understood.

项目总结/摘要新型四环霉素的组合生物合成和代谢研究蒽环类抗生素是天然产物抗生素，其是用于治疗癌症的最有效的抗癌药物之一。诊所蒽环类抗生素的生物合成修饰是一种很有前途的策略，以产生新的化学类似物具有不同的抗癌活性和扩大的治疗窗。Elloramycin（ELM，1）是一种抗生素。一种蒽环类抗生素，由橄榄色链霉菌Tü 2353产生，具有四环埃洛霉素酮糖苷配基和附加的8-O-2，3，4-三-O-甲基-α-L-鼠李糖。埃洛霉素类和四环霉素类由于DNA拓扑异构酶II的抑制，中药表现出轻度的抗增殖活性。此外，委员会认为，脱氧糖部分的优化对于改善细胞渗透、效力、清除和类似物的代谢。然而，我们仍然没有完全理解的结构-活性-关系的中药脱氧糖抗增殖活性及药物代谢。此外，尽管制备了>20 ELM的不同类似物，这些类似物的SAR尚未在一组癌症中进行严格评估。细胞系我们的长期目标是生产具有改善的抗增殖活性的蒽环类药物。到为了促进类似物的生产，我们将利用S. lividans（cos 16 F4）表达系统，以改变通过组合生物合成的四环素脱氧糖部分。这一综合平台将促进用于人类癌症的新药先导物的开发。建议的总体目标研究有三个方面：（1）合成新的中药衍生物，（2）验证抗癌活性， TCM，和（3）开发S9组分测定法，以评价1和最具活性类似物的代谢。我们假设是（1）ElmGT将是“底物柔性的”，足以转移预期的TDP-脱氧糖 8-去甲基-四环霉素C的供体，（2）附加的脱氧糖将通过以下方式改变抗癌活性：与拓扑异构酶II和DNA结合，以及（3）它将通过与肝细胞的差异结合来改变药物代谢细胞色素P450我们的具体目标将测试这些假设：（目标1）我们将异源表达“糖质粒”。通过改变TCM的脱氧糖部分，（目的2）我们将评估新的中药类似物的抗增殖活性，（目的3）我们将开发一种S9级分测定法，以确定新的中药类似物的抗增殖活性。中药类似物的代谢谱。基本原理是“底物柔性”糖基转移酶ElmGT，应该接受新的TDP-脱氧糖供体。这一贡献意义重大，因为预期生产平台将更充分地探索中药类似物的化学空间。此外，这项研究是创新，因为我们正在研究脱氧糖修饰对抗肿瘤活性的影响，四环素的代谢，药物性质目前还不清楚。

项目成果

期刊论文数量（5）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

A BioBricks Metabolic Engineering Platform for the Biosynthesis of Anthracyclinones in Streptomyces coelicolor.

DOI：
10.1021/acssynbio.2c00498
发表时间：
2022-12-16
期刊：
ACS SYNTHETIC BIOLOGY
影响因子：
4.7
作者：
Wang, Rongbin;Nguyen, Jennifer;Hecht, Jacob;Schwartz, Nora;Brown, Katelyn, V;Ponomareva, Larissa, V;Niemczura, Magdalena;van Dissel, Dino;van Wezel, Gilles P.;Thorson, Jon S.;Metsa-Ketela, Mikko;Shaaban, Khaled A.;Nybo, S. Eric
通讯作者：
Nybo, S. Eric

A BioBricks toolbox for metabolic engineering of the tetracenomycin pathway.