Developing chemoenzymatic strategies, enzymes, and kits for accessible and affordable gangliosides

开发化学酶策略、酶和试剂盒，以获得可获取且负担得起的神经节苷脂

• 批准号: 10080549
• 负责人: Lim Andrew Lee
• 金额: $ 90万
• 依托单位: INTEGRATED MICRO-CHROMATOGRAPHY SYSTEMS, INC.
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-08-01 至 2021-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10080549
• 关键词: Acylation; Address; Animals; Biological; Biological Process; Bioreactors; Body Fluids; Brain; Carbohydrates; Cell Communication; Cells; Ceramides; Chemicals; Clinical Trials; Communities; Complex; Cosmetics; Custom; Development; Diagnostic; Economics; Enzymes; Fatty Acids; Feedback; Fermentation; Food; Future; Ganglioside GM1; Gangliosides; Glycoconjugates; Glycoproteins; Glycosides; Growth; Individual; Infection; Inflammation; Libraries; Lipids; Malignant Neoplasms; Methods; Monosaccharides; Nervous system structure; Neuraminidase; Neuraxis; Neurodegenerative Disorders; Oligosaccharides; Parkinson Disease; Play; Polysaccharides; Price; Procedures; Process; Production; Property; Reaction; Reagent; Reporting; Research; Research Personnel; Research Support; Role; Sampling; Source; Specificity; Structure; Sugar Acids; System; Techniques; Technology; Therapeutic; Therapeutic Uses; Tissues; Variant; base; combat; commercialization; cost; flexibility; glycosylation; glycosyltransferase; improved; milligram; miniaturize; nervous system disorder; neurorestoration; pathogen; practical application; sugar; tool

Project Summary Gangliosides, analogous to biological antennae in cells, are involved in critical roles across multiple biological processes (i.e. pathogen infection, cell-cell communication, inflammation). Neuroprotective and neurorestorative functions of gangliosides have been explored for their therapeutic application in Parkinson’s disease and other neurodegenerative diseases. Despite such important role of gangliosides, there is limited access of gangliosides. Majority of the current commercially available gangliosides are obtained from animal brains, which is likely a concern for future therapeutic applications. Such limited availability of gangliosides is further beset by the diversity based on different sugars and fatty acids. The technologies presented here cover an amalgamation of nearly a decade of research, where complex ganglioside is synthesized in a single reaction vessel followed by a simple purification technique. This biosynthetic tool opens the availability of critically needed gangliosides in research and support its therapeutic uses. In this fast track proposal, we will address some major challenges in transitioning this prototypic one-pot multienzyme (OPME) technology for large scale commercialization. First challenge is manufacturing of starting materials (glycosyltransferases, supplemental enzymes and lactosylsphingosine) to reduce the cost to manufacture gangliosides. Additional improvements of existing glycosyltransferases will further reduce the cost burden to manufacture gangliosides by improving catalytic efficiencies of the enzymes. Lastly, OPME scaleability will be explored for manufacturing the various gangliosides. A study will result in several commercial products, ranging from purified enzymes, miniaturized glycosylation kits and several key gangliosides with high commercial value. GM1 ganglioside has been reported to delay onset of Parkinson disease, and clinical trials will likely require animal-free gangliosides. A miniaturized kit will be provided as a tool for researchers to synthesize their own customized gangliosides. The glycosyltransferases and the OPME kits allow the broad scientific community to access these simple synthetic tools for other oligosaccharides, glycoconjugates, and glycoproteins. By expanding this OPME and lowering the cost of these products, additional information on the biological functions of gangliosides will be gained which will accelerate the development of ganglioside-based diagnostics and therapeutics. A positive feedback loop, where lower cost will increase purchase volume, which will feed additional studies and will lead to the further growth of the market for the glycosyltransferases, OPME kits, and highly pure, animal-free gangliosides.

项目摘要 在细胞中类似于生物触角的神经节剂参与多种生物学的关键作用 过程（即病原体感染，细胞传播，炎症）。神经保护性和神经训练 在帕金森氏病和其他 神经退行性疾病。尽管神经毒剂的作用如此重要，但神经节剂的通道有限。 大多数当前可商购的神经毒剂都是从动物大脑中获得的，这很可能是 关注未来的治疗应用。这种有限的神经苷可用性被 基于不同糖和脂肪酸的多样性。这里介绍的技术涵盖了 将近十年的研究，在单个反应容器中合成复杂的神经苷，然后是 一种简单的纯化技术。这种生物合成工具打开了在 研究和支持其治疗用途。在这个快速提案中，我们将解决一些主要挑战 过渡这项原型单锅多酶（OPME）技术，用于大规模商业化。第一的 挑战是制造起始材料（糖基转移酶，补充酶和 乳糖基肾上腺素），以降低生产神经节苷脂的成本。现有的其他改进 糖基转移酶将进一步降低成本燃烧，从 酶的效率。最后，将探索OPME缩放性以制造各种 神经节。一项研究将导致几种商业产品，从纯化的酶，小型化酶 据报道GM1神经节苷脂 为了延迟帕金森氏病的发作，临床试验可能需要无动物的神经节苷脂。小型化 套件将作为研究人员合成自己定制的神经毒剂的工具。 糖基转移酶和OPME套件使广泛的科学界可以访问这些简单的合成 其他寡糖，糖缀合物和糖蛋白的工具。通过扩展此OPME并降低 这些产品的成本，将获得有关神经节苷脂生物功能的其他信息，将获得 加速基于神经节苷脂的诊断和治疗的发展。一个积极的反馈循环，其中 较低的成本将增加购买量，这将提供更多的研究，并将导致进一步的增长 糖基转移酶，OPME套件和高度纯净的无动物神经节苷脂的市场。