Enzymatic methods for assembly of nucleic acid therapeutic agents

修饰的核酸已成为预防和治疗一系列疾病（如Inclisiran，一种革命性的降胆固醇药物）的一种强有力的新型治疗剂。150多项临床试验也正在进行中，主要类型的核酸药物，反义寡核苷酸（ASOs）或小干扰rna (siRNA)，用于癌症，心血管，神经退行性疾病和传染病等。ASOs是短单链核酸（寡核苷酸），与信使RNA （mRNA）杂交，通过阻断核糖体的进展或通过募集核糖核酸酶（RNaseH）诱导mRNA的切割来调节基因表达。siRNA是短的双链RNA分子，与细胞中的特定蛋白质结合形成复合体（RISC），可以切割互补的mRNA靶标。尽管具有巨大的潜力，但这些“灵丹妙药”的生产极具挑战性，因为需要进行广泛的修饰以避免核酸酶降解并提高对mRNA靶标的亲和力。此外，它们的细胞摄取可能会有问题，这通常需要与其他分子结合以帮助传递。目前固相合成主要用于ASOs和sirna的合成。然而，这需要大量昂贵的前体（单体）、有毒试剂、有害溶剂，而且很难扩大规模，这使得治疗成本高昂，阻碍了新药的开发。在这个项目中，我们将开发更可持续和可扩展的酶方法（工业生物技术），在温和的条件下，在水中，利用更良性的酶和可再生的前体，提供必需的ASO和siRNA药物。（详情见申请表格）。

CONFIDENTIALModified nucleic acids have emerged as a powerful new class of therapeutic agents for the prevention and treatment of a range of diseases (e.g. Inclisiran, a revolutionary cholesterol lowering drug. Over 150 clinical trials are also ongoing using the main types of nucleic acid drugs, antisense oligonucleotides (ASOs) or small interfering RNAs (siRNA), for cancer, cardiovascular, neurodegenerative and infectious diseases etc. ASOs are short single stranded nucleic acids (oligonucleotides) that hybridise with messenger RNA (mRNA), modulating gene expression by blocking progression of the ribosome or inducing cleavage of the mRNA through recruitment of a ribonuclease enzyme (RNaseH). siRNA are short double stranded RNA molecules that bind to specific proteins in the cell to form a complex (RISC) which can cleave complementary mRNA targets. Despite their massive potential the production of these 'magic bullets' is extremely challenging, as extensive modification is required to evade nuclease degradation and to improve affinity for the mRNA target. In addition, their cellular uptake can be problematic, which often requires conjugation with other molecules to aid delivery. Currently solid phase synthesis is used to assemble ASOs and siRNAs. However, this requires an excess of costly precursors (monomers), toxic reagents, deleterious solvents and is very difficult to scale up, making treatments expensive and preventing development of new drugs. In this project we will develop more sustainable and scalable enzymatic methods (industrial biotechnology) to deliver essential ASO and siRNA medicines under mild conditions, in water, utilising more benign enzymes and renewable precursors. (see application form for further details).