The quality of peptide APIs is influenced by multiple critical parameters during analytical and process development, and routine manufacturing. In this context the application of state-of-the-art analytical technologies is crucial. The presentation will highlight the most critical success factors from an analytical point of view. This includes a thorough development of analytical procedures, a sound analytical support of the process development, as well as a specific and efficient in-process control strategy for each manufacturing stage. Special focus is laid upon the assurance of the quality of starting materials, a further key element of a holistic quality control strategy for peptide APIs.

The aim of this talk is to highlight the potential value of an increased use of NMR spectroscopy for studies of peptides and peptide impurities. A couple of case studies will be presented including structural characterisation of peptides, quantitative and structural analysis of small molecule and peptide impurities in peptide samples as well as the effectiveness of NMR for comparison of different peptides batches.

The lecture presents the mass spectrometry as a very powerful “tool” of modern analytical chemistry in hands of chemists / scientists performing research or just routine quality control testing of peptides and/or peptoids. The proper use of terminology, interpretation of MS data, main components of mass spectrometers, general LC-MS methodology, with the main emphasis on the most important ionization techniques optimal for MS analysis of these compounds is presented.

The concept of UV Spectral Purity is a well-established technique utilized by the pharmaceutical industry to confirm specificity of HPLC analytical methods for small molecules. It relies on the differences in UV absorbance spectrum between API and impurities. This approach cannot be applied to the peptide industry due to the comparable UV spectrums between the API and the peptidic impurities. The focus of the talk will be on the use of mass spectral techniques to assure that the analytical HPLC methods developed to define API purity presents an appropriate specificity as defined by ICH Q2 (R1). The discussion will focus on the use of mass spectroscopy to determine purity method specificity. The presentation will define instances where MS can support method specificity. Conversely, it will present solutions for impurities that are not correctly defined by MS. The approach to peak purity adopted by Bachem Americas during method development and method pre-validation will be described in detail.

The definition of an oligonucleotide therapeutic has greatly expanded in the past few years with the introduction of novel technologies such as messenger RNAs and CRISPR/CAS9 systems. Furthermore, even the more established oligonucleotide modalities (antisense, siRNA, aptamers, microRNA, etc.) have seen a dramatic increase in the complexity of drug candidates being promoted into preclinical and clinical development. These oligonucleotides often have significant chemical modifications requiring specialty starting materials as well as technical innovations in process development, analytical chemistry, manufacturing and controls. This places extraordinary demands on both sponsor companies and contract service organizations to meet regulatory expectations while ensuring patient safety, often under aggressive timelines. This workshop will present case studies of CMC development of emerging technologies from the perspective of sponsors, CMO and Quality Assurance.

This workshop will provide a high level primer on the use of CRISPR and genome editing technologies in both research and therapeutic applications. An overview of the discovery, preclinical, clinical, CMC, manufacturing and regulatory considerations for CRISPR and genome editing will be discussed. The current challenges and opportunities of CRISPR and genome editing technologies will also be presented.

Topics to be Discussed:

• Creative Chemical Modifications and Synthesis Approaches
• Considerations for Safety, Quality, Regulatory and Off-target Effects
• Advances in Genome Editing TechnologiesDelivery Strategies
• What Can Be Learned from Oligonucleotides and mRNA?
• Applications of CRISPR in Research and Therapeutics

Targeted genome editing with engineered nucleases is increasingly allowing researchers to introduce precise sequence modifications at almost any site within the genome with nucleotide resolution. CRISPR/Cas-mediated genome editing relies on guide RNAs to direct site-specific DNA cleavage mediated by the Cas endonuclease. In this study (Hendel et al., Nat. Biotechnol. 33, 985–989 (2015), doi: 10.1038/nbt.3290), we chemically synthesized 100-nucleotide single guide RNAs (sgRNAs) targeting three different genes (IL2RG, HBB, and CCR5), alongside sgRNA variants containing modified nucleotides at both ends selected for their potential to impact the stability and immunostimulatory properties of nucleic acids. We demonstrate that chemically synthesized sgRNAs can facilitate genome editing in human cell lines when co-delivered with Cas9-encoding plasmid. We further show that sgRNAs containing modified nucleotides dramatically enhance genome editing compared to unmodified sgRNAs. When co-delivering chemically modified sgRNAs and Cas9 mRNA we observe >90% in/del frequencies in human cells lines, ~70% in human primary T cells, and ~40% in CD34+ hematopoietic stem and progenitor cells (HSPCs), despite detecting no activity for unmodified sgRNAs in the latter two cell types. We observe no apparent cellular toxicity and analysis of off-target activities indicate that chemically modified sgRNAs typically retain high specificity. Future studies could investigate a larger variety of chemical modifications, explore different locations of the modifications for rational design of optimized sgRNAs, as well as the mechanism for the enhanced activity of modified sgRNAs. In conclusion, we suggest that next-generation chemically modified sgRNAs have the potential to substantially improve a wide array of CRISPR/Cas biotechnological and therapeutic applications.

Achieving high editing efficiencies in CRISPR therapeutic applications while maintaining consistency remains a significant challenge. Traditional methods for generating guide RNAs yield molecules of inconsistent length and quality that affect genome editing efficiency. We demonstrate that synthetic sgRNA produces consistent editing efficiencies superior to two-piece crRNA:tracrRNA complexes and IVT-derived guides.

The first portion of the discussion will include current market trends in both research and development of therapeutics using CRISPR/Cas gene editing technology and briefly touch on Agilent’s solutions for enabling research. The subsequent portion of the discussion will focus on considerations and approaches that Agilent is using for gRNA production and analysis. A case study will be presented including results from process establishment for gRNA production. Come learn how Agilent is helping its customers turn this exciting technology into a therapeutic reality.

Speaker TBA

Peptides are being developed for a diverse set of clinical uses and have a variety of potencies.  Because processes for manufacture of peptides use common platforms, peptides are often prepared in multi-use (non-dedicated) environments.  Multi- use facilities for non-commercial manufacture present several risk assessment issues that are gaining increasing regulatory scrutiny.  Key topics include dedicated vs multiuse, cleaning validation, occupational containment, facilities design, quality aspects of control strategies.

Topics to be Discussed:

• OEL Banding
• Regulatory Requirements for Cleaning Validation
• Multi-use facilities- Design and Support Considerations
• Cleaning Validation
• Risk Assessment and Containment Strategies for Peptide Drug Products

The safe handling of peptides ranging in toxicity and potency from oligonucleotides to peptide hormones requires appropriate assessment of their toxicity and potency and placing into occupational health categories.  This assessment should be linked to appropriate controls and containment.  As the drug progresses through clinical development, developing quantitative assessments for both worker exposure (Occupational Exposure Limits) and quality assurance (Permitted Daily Exposure for establishing cleaning limits to meet the EMA cross-contamination guideline) are required.  Parallel to these quantitative health-based risk assessments should be development of validated methods for measuring the peptides in air and on surfaces.  This talk will cover scientifically supportable and systematic approaches that can be applied to peptide products in multi-purpose facilities in these critical areas to speed these drugs to market.

e presentation will address important topics when a new Risk and Science based cleaning system is designed. Some of these topics include: 1) How can the risk for cross contamination related to cleaning be decreased? 2) What to do in order to be in compliance with the new EU-GMP directives regarding calculation of carryover levels based on Toxicological evaluations?  3) The importance of the cleaning process (and not just cleaning)

Case study of improvements and new construction required to address critical fire department corrections for a growing and operating peptide facility. Emphasis is on real world assessments, calculations, and justifications needed to comply with California Building and Fire Codes; also applicable to IBC jurisdictions. Session will include brief discussion of control areas, H-x occupancies, building separations and other methods to compartmentalize chemical use in compliance with codes.

The Pharmaceutical Supply Chain Initiative Self Assessment Questionnaire or related documents are often used by larger pharmaceutical companies as part of the assessment of contract manufactures. Often, for early stage programs, insufficient data exists for exposure to API resulting in restrictive occupational exposure handling. Additionally, early stage programs are often prepared in multipurpose suites that are not dedicated or purposely built for specific processes, yet are not suitable to accommodate the more restrictive categorization, even in quantities are quite small. For peptides and oligonucleotides, bioavailability via the oral route or absorption through the skin generally is close to zero percent, yet lack of information at early stages of development introduce a safety factor of 100% bioavailable for both routes, effectively adding a 3 to 4 log reduction in exposure. A review of the key areas of the SHE guidance and discussion of actions from both sponsor and contract manufacturer to potentially help form a more accurate safety assessment will be discussed.

This presentation briefly describes the ideas and principles we are working with in order to shorten development timelines as well as develop robust manufacturing processes. The scope of this presentation is to highlight some of the important parameters to be considered within the purification process development of active synthetic peptides, including phase system optimization, robustness, life time and design of high quality resins. Furthermore, the importance of process understanding and process knowledge by a structured and systematic experimental work (DoE) will be discussed. The responses from DoE will be used to demonstrate the limits of a purification process and thus support a Quality by Design approach (QbD). Although the CQAs (Critical Quality Attributes) of the product have the most prominent place when designing a process, it is not sufficient to design a process only with the CQAs in mind. The scope of the quality concept has to be expanded to encompass all quality attributes of a process, which are: Critical quality attributes of the product, yield, throughput (e.g. kg/day), costs, robustness (e.g. scalability, reproducibility, lack of deviations, out specification, out of trends, reprocesses etc.).

The impurities produced in the solid-phase synthesis of peptides are structurally related and typically have similar chromatographic properties, which can lead to difficulties in the preparation of high purity, clinical-grade peptides.  This can be further complicated when the structural features of the peptides themselves leads to degradation during the chromatography process.  This presentation will describe a number of case studies where a judicious choice of media, buffer and isolation technique has been required in order to produce peptides of the appropriate quality.

Production of high purity synthetic peptides can be a challenging effort due to the complexity of the mixtures and a heavy reliance on chromatographic resources.  The “need for speed” in a time of limited resources adds to this challenge.  By coupling the use of internal and external resources, the process can be streamlined while providing learning opportunities along the way.  In this presentation, we report on this strategy which was employed to efficiently move a critical target through the development phase to produce GMP quality material on kilogram scale.

This workshop will focus on quality expectations for oligonucleotide drug products. A number of new guidances have emerged from the FDA and the EMA recently, and this workshop will explore the application of the underlying quality concerns in those guidances to oligonucleotide drug products. The case studies chosen will span early development through marketed products, in all cases maintaining a focus on the oligonucleotide.

Topics to be Discussed:

• Emerging Regulatory Expectations Regarding Quality of Drug Products
• Terminal Sterilization
• Combination Products
• Co-formulated Products
• Complex Drug Products

This workshop will provide a high level primer on the development of messengerRNA (mRNA) therapeutics. An overview of the discovery, preclinical, clinical, CMC, manufacturing and regulatory considerations for mRNA therapeutics will be discussed. The current challenges and opportunities of these complex molecules will also be presented.

Topics to be discussed include:

• The Anatomy of an mRNA
• mRNA Chemistries
• Manufacturing Strategies for mRNA
• Control Strategies for mRNA
• Applications of mRNA as Therapeutics and Vaccines

Speaker TBA

Peptides offer notable advantages as drugs including their high biological activity, high specificity and low toxicity. However, peptides also present challenges in drug development. (1) short half-life because they are quickly broken down by proteolytic enzymes; (2) inadequate drug-like pharmaceutical properties such as solubility and chemical and physical stability (3) the polar, chemical nature of peptides prevent them from getting past physiological barriers or membranes, and (4) need to be parenterally delivered (via injection) because oral administration would lead to their degradation in the digestive tract. This workshop will focus on cutting-edge, novel technologies to optimize peptides as therapeutics. Case studies will illustrate the use of these platform technologies to enhance the pharmaceutic properties of peptides. 

Topics to be Discussed:

• Stabilizing Peptide Backbone
• Half-life Extension Technologies
• Creative Formulation Approaches
• Strategies to Improve Peptide Delivery
• Other Approaches to Improve Peptide Properties

As a drug class, peptides offer exquisite specificity and potency, but also present challenges associated with poor stability and short half-life, manifesting in the need for frequent injections, poor patient compliance, and overall compromised efficacy. This workshop will cover various peptide stapling technologies that were reported to improve the peptide stability, potency as well as cell penetration. Also, a novel peptide engineering strategy that incorporates a serum protein binding motif onto a covalent side-chain staple will be discussed as well as its application to exenatide, GLP1R agonist, to enhance its helicity and as a consequence, its potency and serum half-life.