Main Conference Day 3Thursday, 13 November 2025 - CET (Central European Time, GMT+01:00)

Oligonucleotides therapeutics have a great potential in a wide range of conditions – many of them where no therapy is currently available. A huge number of products have been approved over the last years. Synthetic oligonucleotides are at the interface of small molecules and biologicals and, from a quality point of view, specific considerations apply to this class of therapeutics. The European Medicines Agency EMA has recently published a draft ‘Guideline on the Development and Manufacture of Oligonucleotides’ which will be presented.

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The talk will share the cross-industry perspective developed within the Synthetic Peptide Working Group of the International Consortium for Innovation and Quality in Pharmaceutical Development (IQ Consortium) on selection of Regulatory starting materials for synthetic peptides. The established approach when using solid phase peptide synthesis leads to all protected amino acids and dipeptides being designed regulatory starting materials (RSMs). As the industry strives to reduce carbon footprint and costs related to drug substance manufacturing, newer alternative manufacture strategies (such as fragment-based approaches) could lead to opportunities for enhanced RSM positions. This talk will give an outline on alternative options, include theoretical case studies.

The global deprotection step is the final stage in the upstream process of peptide synthesis. When peptide elongation is performed via solid-phase peptide synthesis (SPPS), this step coincides with resin cleavage. Traditionally, polyfluoroalkyl substances (PFAS), particularly trifluoroacetic acid (TFA), have been the reagents of choice. However, increasing regulatory scrutiny and anticipated usage restrictions necessitate the development of alternative strategies. Here, we present ongoing efforts in our laboratory to identify and implement viable substitutes for TFA.

Peptide therapeutics have experienced a rapid increase in popularity and demand over recent years, necessitating research into advanced monitoring and manufacturing methods. Within Solid-Phase Peptide Synthesis (SPPS) methods, large amounts of solvent and time are required in sequential and repetitive cycles to manufacture peptide therapeutics. By aligning with the FDA directive for Process Analytical Technology (PAT), Raman spectroscopy shows great promise as an on-line and real-time method for improving the efficiency of SPPS across all major steps of the synthesis. Specifically, Raman spectroscopy can be used to monitor residual piperidine concentration during the de-Fmoc wash step, leading to significant solvent and time savings when used in conjunction with novel continuous wash methods. Additionally, Raman spectroscopy can be used to monitor the concentration of Fmoc-Amino Acid in the reaction solution, which may be useful in developing a greater understanding of the kinetics of the amino acid coupling reactions. The work described herein will outline efforts towards greener, cleaner, and more efficient SPPS methods that can be applied on a commercial scale.

The regulatory landscape for synthetic oligonucleotides has significantly evolved over the past few years. Health Authorities are now providing the first guidelines on this topic, reflecting matured expectations for regulatory submissions. This presentation will explore these evolving expectations and guidelines, offering valuable insights into the current regulatory environment and its implications for future submissions.

Discussion of appropriate areas for application of platform data and approaches. Experience interacting with health authorities will be presented in addition to potential future activities and interactions.

This presentation will explore Regulatory CMC insights from recent global filings for AstraZeneca's commercial ASO program. Key focus areas in Health Authority CMC Questions and complexities related to varying regional regulatory requirements will be discussed.

Discovery and production of synthetic peptides has significantly progressed in the last years resulting in the fact that rather complex peptides like GLP-1 analogues, e.g., Semaglutide and similar sequences, are produced in large quantities for commercial consumption. Length, challenging sequences, and modifications require extended efforts to develop a robust process.

We are demonstrating how different tags placed on different positions on a sequence can help to manufacture difficult peptides in high yield and how such tags can be removed in a traceless manner. This approach includes replacing conventional solid support (resin) on the C-terminus with innovative linkers or a small molecule tag leading to tag-assisted liquid-phase peptide synthesis (LPPS) methodology. Each technology has benefits and drawbacks which will be highlighted, to help the process chemist developing an efficient process, keeping also an eye on material efficiency, sustainability and green aspects in general.

Our unique liquid-phase synthesis technology, utilizing anchor compounds, has been refined for processes like GLP-1 suitable for large-scale production. This method allows post-reaction treatment with simple aqueous washing and uses environmentally friendly solvents. It suppresses side reactions, achieving high purity and yield in long sequence and complex peptide synthesis. We will present examples showing reductions in purification load and PMI.

Solid-Phase Oligonucleotide Synthesis (SPOS) relies on large wash volumes between reagent deliveries to prevent reagent mixing, in large part due to the poor flow characteristics, and resulting reagent mixing, arising from the intentional presence of a headspace during column packing. Dynamic Axial Compression (DAC) columns are commonly used in chromatography to eliminate headspace in packed columns, making the application of this technology to SPOS a logical opportunity to reduce solvent consumption. This presentation will describe Ionis' successful work to scale DAC column operation up to multi-kilo scale production and reduce total solvent wash consumption by ~25%, with no negative impact on yield or purity.

Until recently, interpretation of the Quality Working Party Questions and Answers on API Mix as the only available guidance drove the industry toward energy-intensive lyophilization as the preferred isolation format for highly water-soluble oligonucleotide APIs. The July 2024 EMA draft Guideline on the Development and Manufacture of Oligonucleotides opened the door to registration of oligonucleotide API in solution as an active substance rather than a drug product intermediate. While the solution option affords advantages for sustainability, scale, and supply chain flexibility it also requires a microbial control strategy more typical of biological than synthetic processes. This presentation will propose a robust microbiological control strategy for a generic oligonucleotide purification process, highlighting the appropriate principles of Annex 1 applied to such a non-sterile manufacturing process.

Health authorities are increasingly emphasizing the need for rigorous risk assessments focusing on the formation and presence of nitrosamine impurities in peptide Active Pharmaceutical Ingredients (APIs) produced via synthetic methods. While clear assessment protocols, informed by structure-activity relationship-based methods have been defined for small molecule drug substances, equivalent guidance for synthetic peptides is lacking. This gap in guidance presents significant challenges, including control strategy justification and analytical testing expectations. In this presentation, we will explore relevant literature evidence regarding the formation potential and stability of nitroso-impurities in the context of amino acids, peptides, and proteins. Our study focuses on potential vulnerable nitrogens found in synthetic peptide APIs, such as primary and secondary amides and indoles amongst others. We aim to provide critical insights and general conclusions that inform nitrosamine risk assessments for synthetic peptide API manufacturing processes. Attendees will benefit from a deeper understanding of the formation and stability of nitroso-impurities, enabling them to develop more robust assessment and mitigation strategies.

Exploring novel technologies in solid-phase peptide synthesis designed to improve CMC outcomes—enhancing crude purity, reducing solvent use, and enabling precise temperature control. These innovations support robust process development and tech transfer, offering practical solutions for cost-efficient and scalable peptide manufacturing.

This talk will cover design and implementation of an innovative process for GalNAc-conjugated oligonucleotide drug substances, transitioning from a double chromatography approach to a more efficient single chromatography route. It will highlight the integration of Process Analytical Technology (PAT) in various operations to enhance process understanding and optimization for this new route. We will discuss the challenges faced during technology transfer and the implementation in an GMP manufacture through collaboration with a CMO. Key focus will be given to the improvements in process productivity and sustainability, highlighting substantial advancements compared to the previous process.