Toward synthetic chemically defined mRNA for human therapeutics

用于人类治疗的合成化学定义的 mRNA

• 批准号: 10649299
• 负责人: Keith Thomas Gagnon
• 金额: $ 20.55万
• 依托单位: WAKE FOREST UNIVERSITY HEALTH SCIENCES
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-09-15 至 2025-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10649299
• 关键词: Achievement; Address; Antisense Oligonucleotides; Azoles; Bacteria; Biochemical Reaction; Biological; Bypass; COVID-19 vaccine; Cells; Chemicals; Chemistry; DNA; Development; Disease; Double-Stranded RNA; Enzymes; FDA approved; Formulation; Gene Expression; Generations; Genetic; Genetic Diseases; Genetic Medicine; Half-Life; Heart; Human; In Vitro; Length; Libraries; Medicine; Messenger RNA; Methodology; Methods; Minor; Modality; Modeling; Modification; Molecular Biology; Mus; Nucleic Acids; Nucleotides; Open Reading Frames; Patients; Pharmaceutical Preparations; Phase; Plant Resins; Process; Production; Property; Proteins; RNA; RNA chemical synthesis; Reporter; Research; Ribose; Ribosomes; S phase; Safety; Science; Site; Small Interfering RNA; Solid; Technology; Testing; Therapeutic; Time; Tissues; Toxin; Translations; Vaccines; Vertebral column; chemical synthesis; design; drug development; gene replacement; gene therapy; immunogenic; in vivo; innovation; lipid nanoparticle; manufacture; nuclease; nucleic acid-based therapeutics; patient safety; pharmacologic; sugar; therapeutic RNA; therapeutic development; therapeutic protein

PROJECT SUMMARY Messenger RNA, or mRNA, and its translation into protein lies at the heart of the central dogma of molecular biology. Converting this basic cellular mechanism into a therapeutic opportunity was the basis of the first two successful COVID-19 vaccines. This technology has the potential to be further advanced into much broader therapeutic modalities, such as a gene replacement medicine for genetic diseases. Currently, mRNA molecules for human therapeutics are generated from biological enzymatic reactions. While this process can create large amounts of material, it suffers from several drawbacks. These include multiple steps in manufacturing, purity, and patient safety. However, the greatest shortcoming is the rapid turnover of mRNA in the body, which severely limits its duration of effect and tunability for a genetic medicine. Unless addressed, this shortcoming will handicap mRNA therapeutics from ever becoming more than a vaccine technology. Chemical modification was the missing ingredient and final piece necessary for the realization of other recently FDA-approved nucleic acid drugs, including antisense oligonucleotides and small interfering RNAs. Chemical modifications enabled nuclease protection, significantly extended drug half-lives, and predictable pharmacological tuning. Likewise, realizing the full potential of mRNA as a human therapeutic will ultimately come down to chemistry. RNA can be chemically synthesized in small fragments. However, no technology exists to easily create long chemically defined translation-competent mRNA molecules. In addition, most of the chemical modifications extensively characterized for their beneficial properties for other nucleic acid therapeutics have not been explored in mRNA research, and certainly not in a therapeutic context. This project proposes to tackle these challenges by generating full-length mRNAs from chemically synthesized fragments, investigating the impact of diverse chemical modifications on mRNA translation, and applying new synthetic chemical methods to make longer mRNAs suitable for human therapeutics. The aims of this proposal are to 1) evaluate the impact of specific nucleotide modifications on model mRNA translation in cells and in vitro, 2) assess the compatibility of triazole linkages with mRNA translation and on-resin “click” chemistry for solid-phase chemical synthesis of longer mRNA, and 3) demonstrate long mRNA chemical synthesis and its potential for therapeutic development in cells and in vivo. The results of this focused project should pioneer a paradigm-shifting approach to mRNA therapeutic development and open new possibilities for conferring better control over the drug properties of mRNA.

项目摘要 Messenger RNA或mRNA及其转化为蛋白质的位置位于中央教条的核心 分子生物学。将这种基本的细胞机制转换为治疗机会是基础 在前两种成功的Covid-19疫苗中。这项技术有可能进一步提高 进入更广泛的治疗方式，例如用于遗传疾病的基因替代药物。 当前，人类治疗的mRNA分子是由生物学酶促反应产生的。 尽管此过程可以创建大量材料，但它具有几个缺点。这些包括 制造，纯度和患者安全的多个步骤。但是，最大的缺点是迅速 人体中mRNA的周转率严重限制了其效果持续时间和可线性的通用性 药品。除非解决，否则这种缺点将使mRNA疗法越来越 不仅仅是疫苗技术。 化学修饰是实现其他成分缺失的成分和最后一块 最近，FDA批准的核酸药物，包括反义寡核苷酸和小型干扰 RNA。化学修饰可实现核酸酶保护，大大扩展了药物的半衰期，并且 可预测的药物调整。同样，意识到mRNA作为人类疗法的全部潜力 最终将归结为化学。 RNA可以在小片段中化学合成。但是，没有任何技术可以轻松 创建长期化学定义的翻译能力的mRNA分子。另外，大多数化学物质 其对其他核酸治疗的有益特性的修改广泛特征 在mRNA研究中尚未探索，当然也没有在治疗性情况下进行探讨。这个项目 通过产生化学合成的全长mRNA来应对这些挑战的提议 碎片，调查潜水员化学修饰对mRNA翻译的影响，并应用 新的合成化学方法使MRNA适用于人体治疗。 该提案的目的是1）评估特定核苷酸对模型的影响 细胞中的mRNA翻译和体外，2）评估三唑链接与mRNA的兼容性 用于较长mRNA的固相化学合成的翻译和黄星上的“点击”化学，3） 表现出长长的mRNA化学合成及其在细胞和 体内。这个重点项目的结果应为mRNA治疗的范式转移方法 开发和开放新的可能性，以更好地控制mRNA的药物特性。