Toward synthetic chemically defined mRNA for human therapeutics

用于人类治疗的合成化学定义的 mRNA

基本信息

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
项目类别：
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

项目摘要

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.

项目概要信使 RNA 或 mRNA 及其翻译成蛋白质是信使的中心法则的核心将这种基本的细胞机制转化为治疗机会是基础。前两种成功的 COVID-19 疫苗有进一步发展的潜力。更广泛的治疗方式，例如用于遗传疾病的基因替代药物。目前，用于人类治疗的 mRNA 分子是通过生物酶促反应产生的。虽然这个过程可以产生大量材料，但它也存在一些缺陷。制造、纯度和患者安全方面存在多个步骤，但最大的缺点是速度太快。体内 mRNA 的周转，严重限制了其作用的持续时间和遗传的可调性除非得到解决，否则这一缺点将阻碍 mRNA 疗法的发展。不仅仅是疫苗技术。化学修饰是缺失的成分，也是实现其他功能所必需的最后一块。最近 FDA 批准的核酸药物，包括反义寡核苷酸和小干扰 RNA 的化学修饰可以保护核酸酶，显着延长药物半衰期，并且同样，可预测的药理学调整，实现 mRNA 作为人类治疗的全部潜力。最终将归结为化学。 RNA 可以通过化学方法合成小片段，但是目前还没有技术可以轻松合成。产生长的化学定义的具有翻译能力的 mRNA 分子此外，大多数化学物质。主要特征在于其对其他核酸疗法有益的修饰尚未在 mRNA 研究中进行探索，当然也没有在治疗背景下进行探索。建议通过化学合成生成全长 mRNA 来应对这些挑战片段，研究不同化学修饰对 mRNA 翻译的影响，并应用新的合成化学方法使更长的 mRNA 适合人类治疗。该提案的目的是1）评估特定核苷酸修饰对模型的影响细胞内和体外的 mRNA 翻译，2) 评估三唑键与 mRNA 的相容性用于较长 mRNA 固相化学合成的翻译和树脂上“点击”化学，以及 3) 证明长 mRNA 化学合成及其在细胞和体内治疗开发的潜力这一重点项目的结果应该会开创一种 mRNA 疗法的范式转变方法。开发并为更好地控制 mRNA 的药物特性开辟了新的可能性。