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.

项目总结 信使核糖核酸，或信使核糖核酸，及其到蛋白质的翻译，位于 分子生物学。将这种基本的细胞机制转化为治疗机会是基础 头两种成功的新冠肺炎疫苗中的一种。这项技术有进一步发展的潜力。 转化为更广泛的治疗方式，例如针对遗传病的基因替代药物。 目前，用于人类治疗的信使核糖核酸分子是通过生物酶反应产生的。 虽然这个过程可以产生大量的材料，但它也有几个缺点。这些措施包括 在制造、纯度和患者安全方面的多个步骤。然而，最大的缺点是速度太快 体内信使核糖核酸的周转，这严重限制了它的作用持续时间和基因的可调性。 医药。除非解决这个问题，否则这一缺陷将阻碍信使核糖核酸疗法的形成。 不仅仅是疫苗技术。 化学修饰是缺失的成分，也是实现其他 最近FDA批准的核酸药物，包括反义寡核苷酸和小干扰 RNA。化学修饰能够保护核酸酶，显著延长药物的半衰期，以及 可预见的药理调整。同样，实现信使核糖核酸作为人类治疗药物的全部潜力 最终将归结为化学作用。 核糖核酸可以用化学方法合成成小片段。然而，没有一种技术可以轻松地 产生长的化学定义的具有翻译能力的信使核糖核酸分子。此外，大多数化学物质 被广泛表征为有益于其他核酸疗法的修饰 还没有在信使核糖核酸研究中被探索，当然也不是在治疗的背景下。这个项目 建议通过从化学合成的RNA中产生全长的mRNA来应对这些挑战 片段，研究不同的化学修饰对mRNA翻译的影响，并应用 新的合成化学方法使更长的mRNAs适合于人类治疗。 本提案的目的是：1)评估特定核苷酸修饰对模型的影响 信使核糖核酸在细胞和体外的翻译，2)评估三氮唑键与信使核糖核酸的配伍 翻译和树脂“点击”化学用于固相化学合成较长的信使核糖核酸，和3) 展示长信使核糖核酸的化学合成及其在细胞和肿瘤治疗开发中的潜力 活着。这一重点项目的结果应该是基因治疗范式转换的先驱。 开发并为更好地控制信使核糖核酸的药物特性开辟了新的可能性。