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（由申请方提供）：念珠藻素是一类结构多样的聚酮化合物/非核糖体肽天然产物，具有强效抗癌活性。事实上，念珠藻素合成类似物已被证明有希望治疗铂耐药卵巢癌。尽管有这种令人印象深刻的活性，但由于与治疗相关的临床上显著的周围神经病变，念珠藻素发展为有益的癌症化疗剂受到了影响。然而，这些天然产物的有前途的治疗谱促使Alluvium Biosciences继续发现和开发可以进入临床评估的念珠藻素化合物。为此，Alluvium目前正在开发一种高通量的固相化学酶生产技术，以获得结构多样的念珠藻素化合物，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个化学片段或单位的可用性，它们在固相上组装，随后通过生物催化转化成成熟的念珠藻素化合物。所需的单元中的三个是可商购的或可以通过简单的合成方法容易地获得，而第四个单元，4-羟基苯基辛烯酸，目前必须通过定制的合成策略产生，这导致低的总产率和高的每种化合物的成本。为了增加其念珠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、高产量的4-羟基苯基辛烯酸生产解决方案。因此，在第一阶段SBIR提案中，Alluvium Biosciences将开发一种新的生物合成技术，该技术采用细菌发酵来生产这种关键的念珠藻素聚酮中间体。具体而言，所提出的策略旨在设计和工程化生物合成途径，包括已知的念珠藻素生物合成酶，其将指导在E. coli细菌宿主。在构建工程化生物合成途径并将其转移到细菌宿主中之后，将建立用于产生所需念珠藻素中间体的发酵条件。一旦概念验证得到证实，第二阶段的研究工作将集中在进一步完善基因工程菌株和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生物合成生产化学上多样化的4-羟基苯基辛烯酸类似物的战略。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用作通过Alluvium的固相化学酶促技术可规模化生产所需念珠藻素化合物的起始材料。 公共卫生相关性：癌症代表了一个重大的全球人类健康问题，证明了大量的研究投资，发现和开发新的治疗方法。隐藻素是一种已知的、有效的抗癌化合物，由于无法忍受的副作用而从临床试验中退出。这项拟议的研究旨在帮助开发一种新技术，用于快速生成可能显示更少副作用的念珠藻素类似物，从而使医生能够在对抗这种致命疾病的斗争中使用念珠藻素。