A novel biosynthetic strategy for the production of a key cryptophycin precursor

用于生产关键隐藻素前体的新型生物合成策略

基本信息

批准号：
7998692
负责人：
Jeffrey David Kittendorf
金额：
$ 11.5万
依托单位：
ALLUVIUM BIOSCIENCES, INC.
依托单位国家：
美国
项目类别：
财政年份：
2010
资助国家：
美国
起止时间：
2010-09-24 至 2012-02-25
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7998692
关键词：
Accounting Acids Adverse effects American Cancer Society Amino Acids Anabolism Area Bacterial Chromosomes Biological Factors Biological Sciences Biotechnology Cause of Death Cells Cessation of life Chemical Structure Chemicals Cloning Collaborations Colon Complex Cryptophycin Custom Cyanobacterium Development Development Plans Diagnosis Disease Drops Economics Engineering Enzymes Escherichia coli Fermentation Future Generations Genes Harvest Health Heart Diseases Human Individual Investments Knowledge Licensing Malignant Neoplasms Malignant neoplasm of ovary Mediating Methodology Methods Michigan Operon Pathway interactions Peptides Peripheral Nervous System Diseases Pharmacologic Substance Phase Physicians Platinum Prevention Production Property Prostate Research Resistance Small Business Innovation Research Grant Solid Solid Neoplasm Solutions Source Specificity Staging Synthesis Chemistry System Technology Temperature Therapeutic Time Translating Universities Vendor Work analog anticancer activity chemical synthesis chemotherapeutic agent clinically significant cost cost effective engineering design expression vector follow-up innovation innovative technologies interest large scale production meetings new technology novel programs public health relevance research and development research clinical testing research study scaffold scale up success

项目摘要

DESCRIPTION (provided by applicant): The cryptophycins are a structurally diverse class of polyketide/non-ribosomal peptide natural products that possess potent anticancer activity. In fact, a cryptophycin synthetic analogue has demonstrated promise in treating platinum resistant ovarian cancer. Despite this impressive activity, the development of the cryptophycins into a beneficial cancer chemotherapeutic agent has suffered due to clinically significant peripheral neuropathy that correlates with treatment. However, the promising therapeutic spectrum of these natural products has motivated Alluvium Biosciences to pursue the continued discovery and development of a cryptophycin compound that that can enter clinical evaluation. To this end, Alluvium is currently developing a high throughput, solid-phase chemoenzymatic production technology to enable access to structurally diverse cryptophycin compounds that can be readily screened for desirable pharmacological properties. Key to this effort is the availability of four chemical fragments, or units, which are assembled on solid-phase and subsequently biocatalytically transformed into mature cryptophycin compounds. Three of the required units are either commercially available or can be readily obtained by simple synthetic methods, while the fourth unit, 4- hydroxy phenyloctenoic acid, must currently be generated by custom synthetic strategies that result in low overall yields and high cost per compound. To add value to its cryptophycin chemoenzymatic production technology, Alluvium Biosciences is motivated to develop a low-cost, high yielding solution for the production of 4-hydroxy phenyloctenoic acid. Accordingly, in this Phase I SBIR proposal, Alluvium Biosciences will develop a novel biosynthetic technology that employs bacterial fermentation to produce this critical cryptophycin polyketide intermediate. Specifically, the proposed strategy aims to design and engineer a biosynthetic pathway, comprised of known cryptophycin biosynthetic enzymes, which will direct the biosynthesis of 4- hydroxy phenyloctenoic acid within an E. coli bacterial host. Subsequent to the construction and transfer of the engineered biosynthetic pathway into the bacterial host, fermentation conditions will be established for the production of the desired cryptophycin intermediate. Once proof-of-concept is demonstrated, Phase II research efforts will focus on further refinement of the genetically engineered bacterial strain and transfer of the fermentation technology toward a large-scale production system for the low-cost generation of 4-hydroxy phenyloctenoic acid. Efforts in Phase II will also develop strategies for the biosynthetic production of chemically diverse 4-hydroxy phenyloctenoic acid analogues. Purified 4-hydroxy phenyloctenoic acid, and structural analogues, produced by this fermentation technology will serve as starting material for the scalable production of desirable cryptophycin compounds via Alluvium's solid-phase chemoenzymatic technology. PUBLIC HEALTH RELEVANCE: Cancer represents a significant global human health concern that justifies substantial research investments for the discovery and development of novel treatments. Cryptophycin is a known, potent anti-cancer compound that has been dropped from clinical testing due to intolerable side-effects. This proposed research seeks to aid in the development of a novel technology for the rapid generation of cryptophycin analogues that may display fewer side effects, thereby enabling cryptophycin to be utilized by physicians in the battle against this oft-deadly disease.

描述（由申请人提供）：隐藻素是一类结构多样的聚酮化合物/非核糖体肽天然产物，具有有效的抗癌活性。事实上，隐藻素合成类似物已显示出治疗铂耐药性卵巢癌的前景。尽管具有这一令人印象深刻的活性，但由于与治疗相关的临床显着的周围神经病变，将隐藻素开发成有益的癌症化疗剂却受到了影响。然而，这些天然产品的前景广阔的治疗谱促使 Alluvium Biosciences 继续发现和开发可进入临床评估的隐藻素化合物。为此，Alluvium 目前正在开发一种高通量、固相化学酶生产技术，以便能够获得结构多样的隐藻素化合物，这些化合物可以很容易地筛选出所需的药理学特性。这项工作的关键是四种化学片段或单元的可用性，它们在固相上组装，然后通过生物催化转化为成熟的隐藻素化合物。所需的三个单元要么是商业上可获得的，要么可以通过简单的合成方法轻松获得，而第四个单元，4-羟基苯基辛烯酸，目前必须通过定制合成策略来生成，这导致总体收率低且每种化合物的成本高。为了增加其隐藻素化学酶法生产技术的价值，Alluvium Biosciences 致力于开发一种低成本、高产率的解决方案来生产 4-羟基苯基辛烯酸。因此，在第一阶段 SBIR 提案中，Alluvium Biosciences 将开发一种新型生物合成技术，利用细菌发酵生产这种关键的隐藻素聚酮化合物中间体。具体来说，所提出的策略旨在设计和工程化由已知的隐藻素生物合成酶组成的生物合成途径，该途径将指导大肠杆菌宿主内4-羟基苯基辛烯酸的生物合成。在构建工程生物合成途径并将其转移到细菌宿主中之后，将建立发酵条件以生产所需的隐藻素中间体。一旦概念验证得到证实，第二阶段的研究工作将集中于进一步完善基因工程菌株，并将发酵技术转移到大规模生产系统，以低成本生产 4-羟基苯基辛烯酸。第二阶段的工作还将制定化学多样化的 4-羟基苯基辛烯酸类似物的生物合成生产策略。通过该发酵技术生产的纯化的 4-羟基苯基辛烯酸和结构类似物将作为通过 Alluvium 的固相化学酶技术大规模生产所需隐藻素化合物的起始材料。公共健康相关性：癌症是全球人类健康的一个重大问题，因此需要大量研究投资来发现和开发新的治疗方法。隐藻素是一种已知的强效抗癌化合物，但由于具有无法忍受的副作用，已不再进行临床测试。这项拟议的研究旨在帮助开发一种快速生成隐藻素类似物的新技术，该类似物可能会显示出更少的副作用，从而使医生能够利用隐藻素来对抗这种致命的疾病。