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

• 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
• 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

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）：隐藻蛋白是具有有效抗癌活性的结构多样的聚酮化合物/非核糖体肽天然产物。实际上，隐藻霉素合成类似物在治疗铂抗卵巢癌方面表现出了希望。尽管有这种令人印象深刻的活性，但由于临床上显着的周围神经病变与治疗相关，该隐藻蛋白向有益的癌症化学治疗剂的发展遭受了损失。但是，这些天然产物的有希望的治疗范围促使冲积象生物科学追求可以进入临床评估的隐藻蛋白化合物的持续发现和开发。为此，冲积物目前正在开发高通量，固相化学酶生产技术，以使能够获取结构多样的隐藻蛋白化合物，可以很容易地筛选出理想的药理特性。这项工作的关键是四个化学片段或单位的可用性，这些化学片段或单位被组装在固相上，随后生物催化转化为成熟的隐藻蛋白化合物。所需单元中的三个单元要么可商购，也可以通过简单的合成方法轻松获得，而第四个单元，4-羟基苯二氯辛酸酸，目前必须通过自定义合成策略产生，从而导致整体产量低，每种化合物的成本较高。为了增加其隐霉素化学酶生产技术的价值，促进冲积菌生物科学是为了开发低成本，高屈服的解决方案，用于生产4-羟基苯二氧甲环酸酸。因此，在此I期SBIR提案中，冲积物生物科学将开发一种新型的生物合成技术，该技术采用细菌发酵来产生这种关键的隐霉素聚酮化合物中间体。具体而言，提出的策略旨在设计和设计由已知的隐霉素生物合成酶组成的生物合成途径，该酶将指导大肠杆菌细菌宿主内4-羟基苯二氧甲酸的生物合成。在工程生物合成途径进入细菌宿主之后，将建立发酵条件，以生产所需的隐霉素中间体。一旦证明了概念验证，第二阶段的研究工作将集中于进一步改进基因工程的细菌应变，并将发酵技术转移到大规模生产系统上，以低成本生成4-羟基苯二氧甲酸苯甲酸。第二阶段的努力还将制定化学多样的4-羟基苯二甲酸酯类似物的生物合成生产策略。该发酵技术生产的纯化的4-羟基苯二氧甲酸环烯酸和结构类似物将作为通过冲积物固体化学化学酶技术的可扩展生产所需的隐藻蛋白化合物的起始材料。 公共卫生相关性：癌症代表了全球人类健康的重要关注，这证明了对新治疗的发现和发展的大量研究投资。隐藻霉素是一种已知的有效抗癌化合物，由于无法忍受的副作用，已从临床测试中删除。这项拟议的研究旨在帮助开发一种新型技术，以快速产生可能显示出更少的副作用的隐藻类似物，从而使医生可以在与这种经常疾病的战斗中使用隐藻霉素。