Lumasiran sodium

2024年11月19日，香港城市大学生物医学工程学系史鹏团队在期刊《Nature Communications》上发表了题为“Single-cell encoded gene silencing for high-throughput combinatorial siRNA screening”的研究论文。在筛选抗癌siRNA鸡尾酒时，团队们测试了1,300多种siRNA组合，针对与肿瘤生长相关的多个基因进行敲低，发现了有效的3种siRNA配方，特别强调了抑制Cyclin D1和survivin的关键作用以及它们的互补目标，以实现协同效应。这种方法使得大规模siRNA筛选所需的时间和成本大幅减少，并且解决了现有方法不允许的siRNA药理学的关键见解。 2024年11月19日，香港城市大学生物医学工程学系史鹏团队在期刊《Nature Communications》上发表了题为“Single-cell encoded gene silencing for high-throughput combinatorial siRNA screening”的研究论文。在筛选抗癌siRNA鸡尾酒时，团队们测试了1,300多种siRNA组合，针对与肿瘤生长相关的多个基因进行敲低，发现了有效的3种siRNA配方， 特别强调了抑制Cyclin D1和survivin的关键作用 以及它们的互补目标 ， 以实现协同效应。这种方法使得大规模siRNA筛选所需的时间和成本大幅减少，并且解决了现有方法不允许的siRNA药理学的关键见解。 https://www-nature-com.libproxy1.nus.edu.sg/articles/s41467-024-53419-7 https://www-nature-com.libproxy1.nus.edu.sg/articles/s41467-024-53419-7 关于小干扰核糖核酸 关于小干扰核糖核酸 01 01 小干扰核糖核酸（siRNA）是短的双链RNA序列，它们在被引入到细胞质后会与目标信使RNA（mRNA）结合，通过一种称为RNA干扰（RNAi）的过程发挥作用，导致目标mRNA断裂/降解以阻止其翻译，从而抑制基因表达。迄今为止，美国食品和药品监督管理局（FDA）仅批准了5种基于siRNA的治疗方法：patisiran、givosiran、lumasiran、inclisiran和vutrisiran。 小干扰核糖核酸（siRNA）是短的双链RNA序列，它们在被引入到细胞质后会与目标信使RNA（mRNA）结合，通过一种称为RNA干扰（RNAi）的过程发挥作用，导致目标mRNA断裂/降解以阻止其翻译，从而抑制基因表达。迄今为止，美国食品和药品监督管理局（FDA）仅批准了5种基于siRNA的治疗方法：patisiran、givosiran、lumasiran、inclisiran和vutrisiran。 这项工作旨在通过开发一种高通量技术，以大规模多重、定量和系统的方式筛选基于siRNA的鸡尾酒疗法，来满足未得到满足的需求。团队识别了最有效的3种siRNA组合作为潜在的抗癌疗法，强调抑制Cyclin D1和survivin的关键作用，以及它们的互补基因靶标，以实现协同合作。 这项工作旨在通过开发一种高通量技术，以大规模多重、定量和系统的方式筛选基于siRNA的鸡尾酒疗法，来满足未得到满足的需求。 团队识别了最有效的3种siRNA组合作为潜在的抗癌疗法， 强调抑制Cyclin D1和survivin的关键作用，以及它们的互补基因靶标，以实现协同合作。 组合siRNA鸡尾酒的筛选 组合siRNA鸡尾酒的筛选 02 02 多个siRNA在基因沉默方面，比单一siRNA更有效。随着siRNA转染沉默的基因数量增加，从1个到最多5个siRNA，细胞凋亡分别增加了25.80 ± 3.21%，34.10 ± 1.64%，45.12 ± 6.37%，52.45 ± 5.97%和55.35 ± 10.04%（平均值±标准差）。同样，细胞增殖也与siRNA数量呈负相关，随着siRNA转染从1增加到5，细胞增殖抑制了16.67 ± 1.41%，24.71 ± 1.06%，30.40 ± 5.16%，40.34 ± 2.39%和40.64 ± 4.61%（平均值±标准差）。这些发现表明，多个协同siRNA在诱导期望的基因抑制和相关细胞反应方面更有效。当同时转染4个或更多siRNA时，只有边际的抑制效果增加，这表明如果用这6个选定的基因进行癌症治疗，可能存在用于鸡尾酒治疗的最佳siRNA数量。 多个siRNA在基因沉默方面，比单一siRNA更有效。随着siRNA转染沉默的基因数量增加，从1个到最多5个siRNA，细胞凋亡分别增加了25.80 ± 3.21%，34.10 ± 1.64%，45.12 ± 6.37%，52.45 ± 5.97%和55.35 ± 10.04%（平均值±标准差）。同样，细胞增殖也与siRNA数量呈负相关，随着siRNA转染从1增加到5，细胞增殖抑制了16.67 ± 1.41%，24.71 ± 1.06%，30.40 ± 5.16%，40.34 ± 2.39%和40.64 ± 4.61%（平均值±标准差）。这些发现表明， 多个协同siRNA在诱导期望的基因抑制和相关细胞反应方面更有效。 当同时转染4个或更多siRNA时，只有边际的抑制效果增加，这表明如果用这6个选定的基因进行癌症治疗，可能存在用于鸡尾酒治疗的最佳siRNA数量。 siRNA鸡尾酒的组合筛选，以评估抗肿瘤效果。 siRNA鸡尾酒的组合筛选，以评估抗肿瘤效果。 扩展大规模筛选 扩展大规模筛选 03 03 尽管不同Knock-combos的凋亡水平有所不同，但可以清楚地观察到多途径敲低，比仅针对单一途径更有效。这种协同效应在3个siRNA组合中表现得尤为明显，这与团队在之前小规模筛选中的观察结果一致，并强调了高通量组合siRNA筛选创新的重要性和优势。对于所有1,315个siRNA Knock-combos，团队使用K-means聚类分析发现了两个不同的聚类，其中一个显示出明显更好的诱导凋亡效果。团队得出了一个相对比例，以显示每个聚类中单个siRNA的富集情况（聚类1、2），显示Cyclin D1、survivin、HER2、Hsp 27、UPAR和GSK 3a在凋亡促进聚类（聚类1）中的前6个主导作用，这些在前2个凋亡促进Knock-combos中得到了很好的体现。作为枢纽基因，HER2或Cyclin D1显示出与其它基因最多的连接，表明针对补充基因的siRNA的重要贡献。另一方面，uPAR基因似乎是许多主要siRNA的最佳沉默伙伴，无论是针对survivin还是HER2。这些筛选结果为构建用于抗肿瘤目的的组合siRNA治疗，提供了宝贵的见解。 尽管不同Knock-combos的凋亡水平有所不同，但可以清楚地观察到多途径敲低，比仅针对单一途径更有效。这种协同效应在3个siRNA组合中表现得尤为明显，这与团队在之前小规模筛选中的观察结果一致，并强调了高通量组合siRNA筛选创新的重要性和优势。对于所有1,315个siRNA Knock-combos，团队使用K-means聚类分析发现了两个不同的聚类，其中一个显示出明显更好的诱导凋亡效果。团队得出了一个相对比例，以显示每个聚类中单个siRNA的富集情况（聚类1、2），显示Cyclin D1、survivin、HER2、Hsp 27、UPAR和GSK 3a在凋亡促进聚类（聚类1）中的前6个主导作用，这些在前2个凋亡促进Knock-combos中得到了很好的体现。作为枢纽基因，HER2或Cyclin D1显示出与其它基因最多的连接，表明针对补充基因的siRNA的重要贡献。另一方面，uPAR基因似乎是许多主要siRNA的最佳沉默伙伴，无论是针对survivin还是HER2。这些筛选结果为构建用于抗肿瘤目的的组合siRNA治疗，提供了宝贵的见解。 针对20个基因的由1个、2个或3个siRNA组成的1,315种Knock-combos的扩展筛选。 针对20个基因的由1个、2个或3个siRNA组成的1,315种Knock-combos的扩展筛选。 总结 总结 04 04 1. 本研究开发了一种高通量技术，使用对近红外光和磁场有响应的多层次功能化纳米/微复合载体（Knock-beads），筛选基于siRNA的鸡尾酒疗法，。 1. 本研究开发了一种高通量技术，使用对近红外光和磁场有响应的多层次功能化纳米/微复合载体（Knock-beads），筛选基于siRNA的鸡尾酒疗法，。 2. 该技术通过光穿孔和PDDA层的光热效应实现siRNA的装载和释放，并通过金纳米棒精确控制激光能量。微尺度Fe3O4珠子作为Knock-beads的主干，便于后续成像解码，且不太可能被细胞内吞。通过磁固定、化学交联和尺寸限制，确保Knock-beads与细胞稳定结合，实现单细胞基础的编码和解码。 2. 该技术通过光穿孔和PDDA层的光热效应实现siRNA的装载和释放，并通过金纳米棒精确控制激光能量。微尺度Fe3O4珠子作为Knock-beads的主干，便于后续成像解码，且不太可能被细胞内吞。通过磁固定、化学交联和尺寸限制，确保Knock-beads与细胞稳定结合，实现单细胞基础的编码和解码。 3. 该技术能够显著提高siRNA组合筛选的通量，通过光谱编码方法实现更高级别的多路复用。 3. 该技术能够显著提高siRNA组合筛选的通量，通过光谱编码方法实现更高级别的多路复用。 4. 该研究为siRNA鸡尾酒疗法的开发提供了新的方向，有助于加速siRNA基础治疗的开发和临床应用。 4. 该研究为siRNA鸡尾酒疗法的开发提供了新的方向，有助于加速siRNA基础治疗的开发和临床应用。 参考资料： 参考资料： 1.Fire, A. et al. Potent and specific genetic interference by double-stranded RNA in. Nature 391, 806–811 (1998). 1.Fire, A. et al. Potent and specific genetic interference by double-stranded RNA in. Nature 391, 806–811 (1998). 2.Hamilton, A. J. et al. A species of small antisense RNA in posttranscriptional gene silencing in plants. Science 286, 950–952 (1999). 2.Hamilton, A. J. et al. A species of small antisense RNA in posttranscriptional gene silencing in plants. Science 286, 950–952 (1999).

—Becomes first company to showcase four routes of synthesis for approved siRNA therapeutic asset— —Joint poster with Bachem demonstrates superiority of Company’s double-stranded RNA ligases compared to wild-type enzymes— —Management to host conference call today at 4:30 pm EST to discuss data— REDWOOD CITY, Calif., Nov. 14, 2024 (GLOBE NEWSWIRE) -- Codexis, Inc. (NASDAQ: CDXS), a leading provider of enzymatic solutions for efficient and scalable therapeutics manufacturing, today announced data from three presentations at the TIDES Europe annual meeting being held November 12-14, 2024, in Hamburg, Germany. The data demonstrate the Company’s rapid advancement of its Enzyme Catalyzed Oligonucleotide (ECO) Synthesis™ manufacturing platform and establish Codexis’ position at the forefront of enzymatic synthesis technology to enable to ongoing expansion of RNAi therapeutics. Codexis Demonstrates First-ever Enzymatic Synthesis of Approved siRNA Therapeutic During an oral Spotlight Presentation, Codexis unveiled the successful end-to-end enzymatic synthesis of an entire approved siRNA therapeutic asset, inclisiran. Codexis enzymatically synthesized the full-length sense and antisense strands of the molecule, including the enzymatic incorporation of a tissue-targeting moiety to the sense strand. To date, this process has only been completed utilizing phosphoramidite chemistry, a process that involves the use of harsh chemical conditions and vast amounts of toxic organic solvents. By contrast, Codexis’ ECO Synthesis manufacturing platform operates under milder, aqueous conditions, that improves product quality and dramatically decreases chemical waste production. In addition to this fully enzymatic route of synthesis, the Company demonstrated similar outcomes utilizing three routes of enzymatic ligation to produce the siRNA therapeutic asset, combining oligonucleotide fragments made by sequential enzymatic synthesis and traditional phosphoramidite chemistry. Key data from the presentation include: Achieved incorporation efficiency of >98% during sequential enzymatic oligo synthesisSuccessfully attached the tri-GalNAc tissue-targeting moiety by enzymatic ligationObtained full-length oligonucleotides of equal quality and yields, using ligation of short fragments made with enzymes or by traditional phosphoramidite chemistry Now that Codexis has successfully achieved this unprecedented milestone, the Company will continue to optimize its process for robustness, scaled-up quantities and improved purity with the goal of providing customers with siRNA material of comparable or better quality to phosphoramidite chemistry. The Company anticipates ramping up manufacturing of siRNA in quantities for preclinical testing following the successful build out of its ECO Synthesis Innovation Lab at the end of this year. Codexis Double-stranded RNA Ligase Demonstrates Superior Performance to Wild-type Enzymes Two additional presentations focused on results of direct comparisons of Codexis’ engineered double-stranded RNA (dsRNA) ligases and wild-type (WT) enzymes when used to combine short oligonucleotide fragments to synthesize full-length siRNA therapeutic compounds. In a joint poster with Bachem, one of the world’s leading CDMOs in oligonucleotide manufacturing, the data provided compelling external validation of the superior performance of Codexis ligases over existing wild-type enzymes in use today. Codexis enzymes demonstrated superior performance over wild-type enzymes across both volumetric productivity and substrate versatility. These dsRNA ligases outperformed on multiple substrate designs and enabled a higher conversion rate of oligonucleotide fragments into siRNA at increased concentrations of raw materials. In a separate TIDES Talk presentation, Codexis demonstrated improved performance of its dsRNA ligase over WT enzymes based on real customer case studies executed through the Company’s RNA Ligase Screening and Optimization Services, launched in May 2024. Codexis’ engineered ligases delivered robust, in-process performance, including higher substrate loading, faster reaction times and improved conversation at elevated temperatures. These data demonstrate the Company’s ability to accelerate delivery of lead ligase variants to customers and optimize process conditions for a customer’s specific asset. The slide decks from both presentations as well as the joint poster with Bachem are now available in the Investor Relations section the Codexis corporate website, www.codexis.com/investors. Conference Call and Webcast Codexis management will host a conference call beginning at 4:30 pm Eastern Time on Thursday, November 14, 2024, to discuss the data presented during the conference. The live call can be accessed by dialing 877-705-2976 (domestic) or 201-689-8798 (international). A live webcast to accompany the conference call can be accessed on the Codexis Investor Relations website, where a replay will be available for 90 days. A telephone replay of the call will be available for 48 hours by dialing 877-660-6853 (domestic) or 201-612-7415 (international), access ID #13726635. About RNAi Therapeutics Manufacturing Ribonucleic acid (RNA) as a therapeutic modality has gained tremendous traction in recent years with the growing number of messenger RNA (mRNA) vaccines and small interfering RNA (siRNA) candidates advancing in clinical studies. However, large-scale production of RNA interference (RNAi) therapeutics using traditional chemical synthesis faces complex challenges in nucleic acid quality and quantity, as well as overall economics. With over 450 RNAi therapies currently in clinical development, including more than 40 assets in Phase 2 and Phase 3 clinical trials targeting disease indications impacting millions of patients, RNAi therapeutic demand is projected to outpace current production capabilities by the end of the decade. About the ECO Synthesis Manufacturing Platform Codexis’ proprietary Enzyme Catalyzed Oligonucleotide (ECO) Synthesis™ manufacturing platform is being designed to address scalability and cost limitations by potentially enabling the commercial-scale manufacture of RNAi therapeutics through an enzymatic route. The Company presented groundbreaking data at the TIDES USA 2024 annual meeting demonstrating the enzymatic synthesis of a full-length sense strand of the oligonucleotide lumasiran, a commercially available siRNA therapeutic, as well as shorter sense strand fragments of a second siRNA therapeutic asset, givosiran. The data demonstrate that Codexis consistently achieved coupling efficiency greater than 98%, which is equivalent to what is seen with phosphoramidite chemistry; executed the enzymatic addition of a conjugation moiety to the lumasiran strand; and confirmed the lack of notable impurities typically observed in oligonucleotide synthesis via phosphoramidite chemistry. A recording of the presentation, along with slides and the data press release, can be found on the Codexis corporate website. About Codexis Codexis is a leading provider of enzymatic solutions for efficient and scalable therapeutics manufacturing that leverages its proprietary CodeEvolver® technology platform to discover, develop and enhance novel, high-performance enzymes and other classes of proteins. Codexis enzymes solve for real-world challenges associated with small molecule pharmaceuticals manufacturing and nucleic acid synthesis. The Company is currently developing its proprietary ECO Synthesis™ manufacturing platform to enable the scaled manufacture of RNAi therapeutics through an enzymatic route. Codexis’ unique enzymes can drive improvements such as higher yields, reduced energy usage and waste generation, improved efficiency in manufacturing and greater sensitivity in genomic and diagnostic applications. For more information, visit https://www.codexis.com. Forward-Looking Statements This press release contains forward-looking statements within the meaning of Section 27A of the Securities Act of 1933, as amended, and Section 21E of the Securities Exchange Act of 1934, as amended. In some cases, you can identify forward-looking statements by terminology such as “aim,” “anticipate,” “assume,” “believe,” “contemplate,” “continue,” “could,” “design,” “due,” “estimate,” “expect,” “goal,” “intend,” “may,” “objective,” “plan,” “positioned,” “potential,” “predict,” “seek,” “should,” “suggest,” “target,” “on track,” “will,” “would” and other similar expressions that are predictions of or indicate future events and future trends, or the negative of these terms or other comparable terminology. To the extent that statements contained in this press release are not descriptions of historical facts, they are forward-looking statements reflecting the current beliefs and expectations of management, including, but not limited to, the ability of an enzymatic oligonucleotide synthesis process to complement, replace or improve upon traditional chemical synthesis; the potential of the Company’s ECO Synthesis™ platform and double-stranded RNA (dsRNA) ligase screening and optimization services to meet customers’ needs and to create value for Codexis and its customers by enabling commercial-scale manufacture of RNAi therapeutics; completion of the ECO Synthesis Innovation Lab by the end of 2024, and other anticipated technical and commercial milestones related to the ECO Synthesis™ platform and the dsRNA ligase program, and public announcements related thereto; ability of the Company to obtain new development collaborators on its ECO Synthesis technology; the potential for the Company’s dsRNA ligases to have improved scalability and reduced manufacturing costs compared to wild types or non-ligation methods; potential details and features of the ECO Synthesis™ platform such as it being scalable and able to reduce manufacturing costs, as well as having higher quality, purity and yield, and improved sustainability than existing methods; and the future demand for RNAi therapeutics. You should not place undue reliance on these forward-looking statements because they involve known and unknown risks, uncertainties and other factors that are, in some cases, beyond Codexis’ control and that could materially affect actual results. Factors that could materially affect actual results include, among others: Codexis’ dependence on its licensees and collaborators; if any of its collaborators terminate their development programs under their respective license agreements with Codexis; Codexis may need additional capital in the future in order to expand its business; if Codexis is unable to successfully develop new technology such as its ECO Synthesis™ manufacturing platform and dsRNA ligase; Codexis’ dependence on a limited number of products and customers, and potential adverse effects to Codexis’ business if its customers’ products are not received well in the markets; whether the end markets for Codexis’ customers’ products develop and remain viable; if competitors and potential competitors who have greater resources and experience than Codexis develop products and technologies that make Codexis’ products and technologies obsolete; Codexis’ ability to comply with debt covenants under its loan facility; if Codexis is unable to accurately forecast financial and operational performance; and market and economic conditions may negatively impact Codexis business, financial condition and share price. Additional information about factors that could materially affect actual results can be found in Codexis’ Annual Report on Form 10-K for the year ended December 31, 2023 filed with the Securities and Exchange Commission (“SEC”) on February 28, 2024 and in Codexis’ Quarterly Report on Form 10-Q for the quarter ended September 30, 2024 filed with the SEC on October 31, 2024, including under the caption “Risk Factors,” and in Codexis’ other periodic reports filed with the SEC. Codexis expressly disclaims any intent or obligation to update these forward-looking statements, except as required by law. For More Information Investor ContactCarrie McKim(336) 608-9706ir@codexis.com Media ContactLauren Musto(650) 421-8205media@codexis.com