Daniel O'Reilly, Raymond Furgal, Vignesh Hariharan, Clemens Lochmann, David Cooper, Dimas Echeverria, Anastasia Khvorova
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引用次数: 0
Abstract
Chemically modified short interfering RNAs (siRNAs) unequivocally represent a groundbreaking class of drugs. The deliberate chemical modification of the natural structure has been pivotal to their resounding success. Specific modifications at certain positions bolster their potency, safety, stability, and specificity. In clinical research, 2'-O-methyl and 2'-fluoro are the most used modifications. The effects of a wide range of chemical changes in fully modified siRNAs have not been thoroughly evaluated for tolerability. In this study, we utilized two sequences in a fully modified siRNA to systematically assess the tolerability of single nucleotide backbone and sugar modifications, including deoxyribonucleic acid, 2'-O-(2-methoxyethyl), locked nucleic acid, unlocked nucleic acid, mismatches, butane diol substitution, and butane diol insertion. We synthesized 522 siRNA variants and evaluated their efficacy in vitro. Our findings demonstrate that individual tolerability is significantly influenced by the modification's sequence, pattern, and position, with limited universal principles identifiable from this dataset. The efficacy results are probably driven by the thermodynamic balance defined by a combination of parameters. The framework presented here will serve as a reference dataset to facilitate the expansion of chemical diversity in therapeutic siRNAs.
化学修饰的短干扰rna (sirna)无疑代表了一类突破性的药物。对自然结构进行刻意的化学修饰是他们取得巨大成功的关键。特定位置的特异性修饰增强了它们的效力、安全性、稳定性和特异性。在临床研究中,2'- o -甲基和2'-氟是最常用的修饰。广泛的化学变化对完全修饰的sirna的影响尚未被彻底评估其耐受性。在这项研究中,我们利用一个完全修饰的siRNA中的两个序列,系统地评估了单核苷酸主链和糖修饰的耐受性,包括脱氧核糖核酸、2'- o -(2-甲氧基乙基)、锁定核酸、解锁核酸、错配、丁烷二醇取代和丁烷二醇插入。我们合成了522种siRNA变体,并评估了它们在体外的功效。我们的研究结果表明,个体的耐受性受到修饰的序列、模式和位置的显著影响,从这个数据集中可以识别出有限的普遍原则。效能结果可能是由一系列参数所定义的热力学平衡所驱动的。本文提出的框架将作为一个参考数据集,以促进治疗性sirna化学多样性的扩展。
期刊介绍:
Nucleic Acid Therapeutics is the leading journal in its field focusing on cutting-edge basic research, therapeutic applications, and drug development using nucleic acids or related compounds to alter gene expression. The Journal examines many new approaches for using nucleic acids as therapeutic agents or in modifying nucleic acids for therapeutic purposes including: oligonucleotides, gene modification, aptamers, RNA nanoparticles, and ribozymes.