Chemistry , Manufacturing And Controls Research Articles

Studying the stability of polymer nanoparticles by size exclusion chromatography of radioactive polymers

The properties of nanomedicines will influence how they can deliver drugs to patients reproducibly and effectively. For conventional pharmaceutical products, Chemistry, Manufacturing and Control (CMC) documents require monitoring stability and storage conditions. For nanomedicines, studying these important considerations is hindered by a lack of appropriate methods. In this paper, we show how combining radiolabelling with size exclusion chromatography, using a method called SERP, can inform on the in vitro degradation of polymers. Using nanoparticles composed of biodegradable poly(lactic acid) (PLA) and poly(lactic-co-glycolic acid) (PLGA), we show that SERP is more sensitive than dynamic light scattering (DLS) and nanoparticle tracking analysis (NTA) to detect degradation. We also demonstrate that the properties of the polymer composition and the nature of the aqueous buffer affect polymer degradation. Importantly, we show that minute changes in stability that cannot be detected by DLS and NTA impact the pharmacokinetic of nanoparticles injected in vivo. We believe that SERP might prove a valuable method to document and understand the pharmaceutical quality of polymer nanoparticles.

Supercoiled DNA percentage: A key in-process control of linear DNA template for mRNA drug substance manufacturing

The development of messenger RNA (mRNA) vaccines and therapeutics necessitates the production of high quality in-vitro-transcribed mRNA drug substance with specific critical quality attributes (CQAs), which are closely tied to the uniformity of linear DNA template. The supercoiled plasmid DNA is the precursor to the linear DNA template, and the supercoiled DNA percentage is commonly regarded as a key in-process control (IPC) during the manufacturing of linear DNA template. In this study, we investigate the influence of supercoiled DNA percentage on key mRNA CQAs, including purity, capping efficiency, double-stranded RNA (dsRNA), and distribution of polyA tail. Our findings reveal a significant impact of supercoiled DNA percentage on mRNA purity, and in vitro transcription yield. Notably, we observe that the impact on mRNA purity can be mitigated through Oligo-dT chromatography, alleviating the tight range of DNA supercoiled percentage to some extent. Overall, this study provides valuable insights into IPC strategies for DNA template CMC (Chemistry, Manufacturing, and Controls) and process development for mRNA drug substance.

Science- and Risk-Based Stability Strategies to Support Product Lifecycle Changes.

ICH Q12 asserts that science- and risk-based approaches are applicable to stability studies supporting Chemistry, Manufacturing and Controls (CMC) post-approval changes (PAC) to enable more timely implementation; however, no guidance or specific examples are provided to demonstrate how prior knowledge of the product can inform the risk assessment for the proposed change(s). Ten diverse case studies are presented in this manuscript to demonstrate how science- and risk-based stability strategies were used to support drug substance and product CMC PAC and lifecycle management activities. The accumulated stability knowledge held by original manufacturers of marketed products is substantial, and different elements of this knowledge base were used to assess the risks and impact of the proposed changes for confident change management. This paper provides ways to leverage science- and risk-based stability strategies as part of the post-approval change-management risk-mitigation strategy, which may enable a reduced stability data commitment and/or a reduced reporting category for change implementation.

Further accelerating biologics development from DNA to IND: the journey from COVID-19 to non-COVID-19 programs.

The Coronavirus Disease (COVID-19) pandemic has spurred adoption of revolutionary initiatives by regulatory agencies and pharmaceutical industry worldwide to deliver therapeutic COVID-19 antibodies to patients at unprecedented speed. Among these, timeline of chemistry, manufacturing and control (CMC), which involves process development and manufacturing activities critical for the assurance of product quality and consistency before first-in-human clinical trials, was greatly reduced from typically 12-15months (using clonal materials) to approximately 3months (using non-clonal materials) in multiple cases. In this perspective, we briefly review the acceleration approaches published for therapeutic COVID-19 antibodies and subsequently discuss the applicability of these approaches to achieve investigational new drug (IND) timelines of ≤10months in over 60 COVID-19 and non-COVID-19 programs performed at WuXi Biologics. We are of the view that, with demonstrated product quality and consistency, innovative approaches used for COVID-19 can be widely applied in all disease areas for greater speed to clinic.

Evaluation of the impact of reliance on the regulatory performance in the South African Health Products Regulatory Authority: implications for African regulatory authorities.

The World Health Organization (WHO) advocates the use of reliance practices to enable national regulatory authorities (NRAs) to improve patients' access to medicines. This study considered whether reliance review translates into swifter medicine authorization. Abridged review outcomes were examined for New Chemical Entity (NCE) and generic applications to the South African Health Products Regulatory Authority (SAHPRA) in Chemistry, Manufacturing and Controls (CMC) and clinical/bioequivalence (BE), as well as overall NCE authorization times. SAHPRA NCE CMC review time was 91 days (abridged) vs. 179 days (full), applicant response time was 34 vs. 105 days, respectively, and there was a >2-fold time reduction for abridged vs. full CMC review (125 vs. 284 days). There was a 99-day decrease in clinical approval time through an abridged review (230 vs. 329 days) and a decrease in marketing authorization time for NCE abridged assessment (446 vs. 619 days). SAHPRA review time for generic applications was 97 days (abridged) vs. 191 days (full); applicant response time was 26 days (abridged) vs. 81 days (full) and there was a >2-fold time reduction for CMC and BE abridged vs. full review (122 vs. 272 days). These results clearly support World Health Organization recommendations for the use of reliance-based regulatory review to expedite the worldwide availability of safe, effective and needed medications.

Co-crystallisation and humanisation of an anti-HER2 single-domain antibody as a theranostic tool.

Human epidermal growth factor receptor-2 (HER2) is a well-recognised biomarker associated with 25% of breast cancers. In most cases, early detection and/or treatment correlates with an increased chance of survival. This study, has identified and characterised a highly specific anti-HER2 single-domain antibody (sdAb), NM-02, as a potential theranostic tool. Complete structural description by X-ray crystallography has revealed a non-overlapping epitope with current anti-HER2 antibodies. To reduce the immunogenicity risk, NM-02 underwent a humanisation process and retained wild type-like binding properties. To further de-risk the progression towards chemistry, manufacturing and control (CMC) we performed full developability profiling revealing favourable thermal and physical biochemical 'drug-like' properties. Finally, the application of the lead humanised NM-02 candidate (variant K) for HER2-specific imaging purposes was demonstrated using breast cancer HER2+/BT474 xenograft mice.

Preclinical studies of toxicity and anti-cholangiocarcinoma activity of the standardized capsule formulation of Atractylodes lancea (Thunb.) DC

BackgroundCholangiocarcinoma (CCA), the adenocarcinoma of the biliary duct, is commonly reported in Asia, with the highest incidence in northeastern Thailand. Chemotherapy of CCA has been limited by the lack of effective chemotherapeutic drugs. A series of previous in vitro and in vivo studies support further research and development of Atractylodes lancea (Thunb.) DC. (AL) as a potential candidate for treating CCA as a crude ethanolic extract. In the present study, we evaluated the toxicity and anti-CCA activity of the CMC (Chemistry, Manufacturing, and Control) capsule formulation of the ethanolic rhizome extract of AL (CMC-AL) in animals.MethodsMajor steps included acute, subchronic and chronic toxicity testing in Wistar rats and anti-CCA activity in a CCA-xenografted nude mouse model. The safety of CMC-AL was determined based on the maximum tolerated dose (MTD) and no-observed-adverse-effect level (NOAEL) according to the OECD guideline. The anti-CCA activity of CMC-AL in nude mice was evaluated after transplantation of CL-6 cells to evaluate inhibitory effects on tumor size progression and metastasis and survival time prolongation. Safety assessments included hematology, biochemistry parameters and histopathological examination. Lung metastasis was investigated using VEGF ELISA kit.ResultsAll evaluations confirmed satisfactory pharmaceutical properties of oral formulation and safety profile of the CMC-AL with no overt toxicity up to the MTD and NOAEL of 5,000 and 3,000 mg/kg body weight, respectively. CMC-AL exhibited potent anti-CCA efficacy with regard to inhibitory activity on tumor progression and lung metastasis.ConclusionsCMC-AL is safe and should be further investigated in a clinical trial as a potential therapy for CCA patients.

Survey Outcome on Immunogenicity Risk Assessment Tools for Biotherapeutics: an Insight into Consensus on Methods, Application, and Utility in Drug Development.

A survey conducted by the Therapeutic Product Immunogenicity (TPI) community within the American Association of Pharmaceutical Scientists (AAPS) posed questions to the participants on their immunogenicity risk assessment strategies prior to clinical development. The survey was conducted in 2 phases spanning 5years, and queried information about in silico algorithms and in vitro assay formats for immunogenicity risk assessments and how the data were used to inform early developability effort in discovery, chemistry, manufacturing and control (CMC), and non-clinical stages of development. The key findings representing the trends from a majority of the participants included the use of high throughput in silico algorithms, human immune cell-based assays, and proteomics based outputs, as well as specialized assays when therapeutic mechanism of action could impact risk assessment. Additional insights into the CMC-related risks could also be gathered with the same tools to inform future process development and de-risk critical quality attributes with uncertain and unknown risks. The use of the outputs beyond supporting early development activities was also noted with participants utilizing the risk assessments to drive their clinical strategy and streamline bioanalysis.

Cell identity, count, and viability for critical quality attributes using the Cellaca PLX image cytometer

Abstract Cell and gene therapy is one of the fastest growing fields for cancer therapeutics that heavily relies on robust and consistent instrumentations and technologies to verify and validate the fit-for-purpose critical quality attributes (CQA). Some of the key parameters required for satisfying CMC (Chemistry Manufacturing and Controls) criteria standards for cellular therapeutic products are routinely performed using flow cytometry. In the past decade, image cytometry was developed for cell characterization and cell-based assays but had not yet demonstrated the sensitivity required for surface marker detection. We demonstrate the capability of the Cellaca PLX image cytometry system for surface marker population analysis, GFP and RFP cell viability, and cell health in comparison to flow cytometry. For immunophenotyping, PBMCs were stained with Hoechst/CD3/CD4/CD8, and viability detection of GFP and RFP-containing cell lines was performed by staining with Hoechst/RubyDead. Additionally, apoptosis was induced in Jurkat cells with staurosporine and cells stained with Hoechst/Caspase 3-488/RubyDead. All assays were analyzed on the Cellaca PLX and compared directly to the CytoFLEX. Results show similar surface marker populations: CD3 (76% and 81%, PLX and CytoFLEX, respectively), CD4 (43% and 42%), and CD8 (15% and 17%). Viabilities of RFP and GFP cells were equivalent on both platforms, and population percentages of Hoechst/Caspase 3/RubyDead positive cells were within 5% of each other. Our experiments demonstrate that the Cellaca PLX image cytometer can be of significant value to the Cell and Gene Therapy communities to satisfy several CMC criteria including high-throughput, high sensitivity, and low maintenance.

Abstract 5330: Cell identity, count, and viability for critical quality attributes using the Cellaca PLX image cytometer

Abstract Cell and Gene therapy is one of the fastest growing fields for cancer therapeutics that heavily relies on robust and consistent instrumentations and technologies to verify and validate the critical quality attributes (CQA) that are fit-for-purpose. Some of the key parameters required for satisfying CMC (Chemistry Manufacturing and Controls) criteria standards for cellular therapeutic products are cell identity, count, viability, purity, potency, stability, and microbiological quality, which may be routinely performed using flow cytometry. Some of these critical attributes can be exceedingly trivial for flow cytometric analysis, which can be complex, requires a dedicated user, and may carry a high maintenance cost. In the past decade, affordable image cytometry has been developed for cell characterization and cell-based assays but has not demonstrated the sensitivity required for visualization and analysis of surface markers, cell health, and viability. In this work, we demonstrate the capability of the newly developed Cellaca PLX image cytometry system for population analysis of surface markers, viability of fluorescent protein expressing cells, and cell health in comparison to flow cytometry. For immunophenotyping assays, PBMCs were stained with anti-human CD3/CD4/CD8 antibodies with Hoechst. Additionally, viability detection of GFP and RFP-containing cell lines was performed by staining with RubyDead and Hoechst. Finally, apoptosis was induced in Jurkat cells with staurosporine and stained with Caspase 3-488, RubyDead, and Hoechst. All assays were imaged and analyzed using the Cellaca PLX image cytometer, and the data generated were compared directly to the CytoFlex flow cytometer. Results show similar percentages of surface marker populations: CD3 (76% and 81%, PLX and CytoFlex, respectively), CD4 (43% and 42%), and CD8 (15% and 17%). RFP and GFP-containing cells showed comparable viabilities on Cellaca PLX and CytoFlex, and the population percentages of Hoechst positive cells stained with Caspase 3 and RubyDead were within 5% of each other. Our experiments demonstrate that the Cellaca PLX can be of significant value to the Cell and Gene Therapy communities providing novel image cytometry methods which may satisfy several CMC criteria including high-throughput, high sensitivity, and low maintenance. Citation Format: Carolina Franco Nitta, Mackenzie Pierce, Aiyana Parker, Sopaul Hem, Timothy Smith, Surbhi Ratnani, Marek Dobrowolski, Srinivas Gundimeda, Bo Lin, Dmitry Kuksin, Leo L. Chan. Cell identity, count, and viability for critical quality attributes using the Cellaca PLX image cytometer. [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 5330.

Vaccines development in India: advances, regulation, and challenges.

One of the most significant medical advancements in human history is the development of vaccines. Progress in vaccine development has always been greatly influenced by scientific human innovation. The main objective of vaccine development would be to acquire sufficient evidence of vaccine effectiveness, immunogenicity, safety, and/or quality to support requests for marketing approval. Vaccines are biological products that enhance the body's defenses against infectious diseases. From the first smallpox vaccine to the latest notable coronavirus disease 2019 nasal vaccine, India has come a long way. The development of numerous vaccines, driven by scientific innovation and advancement, combined with researcher's knowledge, has helped to reduce the global burden of disease and mortality rates. The Drugs and Cosmetics Rules of 1945 and the New Drugs and Clinical Trials Rules of 2019 specify the requirements and guidelines for CMC (chemistry, manufacturing, and controls) for all manufactured and imported vaccines, including those against coronavirus infections. This article provides an overview of the regulation pertaining to the development process, registration, and approval procedures for vaccines, particularly in India, along with their brief history.

Developability assessment at early-stage discovery to enable development of antibody-derived therapeutics.

Developability refers to the likelihood that an antibody candidate will become a manufacturable, safe and efficacious drug. Although the safety and efficacy of a drug candidate will be well considered by sponsors and regulatory agencies, developability in the narrow sense can be defined as the likelihood that an antibody candidate will go smoothly through the chemistry, manufacturing and control (CMC) process at a reasonable cost and within a reasonable timeline. Developability in this sense is the focus of this review. To lower the risk that an antibody candidate with poor developability will move to the CMC stage, the candidate's developability-related properties should be screened, assessed and optimized as early as possible. Assessment of developability at the early discovery stage should be performed in a rapid and high-throughput manner while consuming small amounts of testing materials. In addition to monoclonal antibodies, bispecific antibodies, multispecific antibodies and antibody-drug conjugates, as the derivatives of monoclonal antibodies, should also be assessed for developability. Moreover, we propose that the criterion of developability is relative: expected clinical indication, and the dosage and administration route of the antibody could affect this criterion. We also recommend a general screening process during the early discovery stage of antibody-derived therapeutics. With the advance of artificial intelligence-aided prediction of protein structures and features, computational tools can be used to predict, screen and optimize the developability of antibody candidates and greatly reduce the risk of moving a suboptimal candidate to the development stage.

Consideration of quality by design principles in the use of sterilizing grade filters for clarification and final sterile filtration in adeno-associated virus applications

The development of gene therapy applications using recombinant adeno-associated viruses (AAV) as a gene delivery vector has gained significant momentum in recent years. Although there are numerous applications for potential therapies to treat unmet medical needs, the ability to implement robust AAV manufacturing processes continues to be a bottleneck to fully realizing the potential of AAV based therapeutics. This includes the requirement to understand the characteristics (quality attributes) that are critical to product quality and patient safety, and to define these in appropriate detail for regulatory submissions such as chemistry, manufacturing and controls (CMC) information. Sterilizing grade filtration plays an important role in AAV manufacturing, both for drug substance (DS) as well as final drug product (DP). However, to fulfill regulatory expectations for provision of CMC information, it is recommended that the principles of quality by design (QbD) are considered during the early product development stages of the AAV manufacturing process. Both prior knowledge and experimental data should be used to identify potential critical quality attributes (CQAs). Enhanced knowledge should result from increased experience, and ultimately the critical process parameters (CPPs) and critical material attributes (CMAs) that can impact the CQAs can then be defined. The objective is to define a manufacturing control strategy that includes appropriate control of CPPs and CMAs to assure the CQAs of the drug product. Here, the application of QbD principles for sterilizing grade filtration in both DS clarification and DP final fill steps of a typical AAV manufacturing process is described. Both experimental and data review approaches are included to help understand what a typical design space could look like, from which appropriate controls can be implemented. Ultimately, the expectations from regulatory agencies for assurance of product quality and patient safety are aligned with the requirement for a robust manufacturing control strategy to ensure consistent product quality is achieved.

Considerations for the chemistry, manufacturing and Controls (CMC) - quality package for COVID-19 vaccines- interim lessons learnt by the European medicines Agency (EMA)

The European Medicines Agency (EMA) has approved five pandemic COVID-19 vaccines (prior to April 2022) and many others are in the pipeline. The commentary describes how timely approval and rapid manufacturing capacity scale up could be achieved from our perspective.The commentary considers the need for: early, continuous engagement with the regulator for COVID-19 vaccines; understanding key Chemistry, Manufacturing and Controls (CMC) challenges in order to build a successful COVID-19 vaccine CMC dossier; investing in production and testing site readiness for COVID-19 vaccines; CMC Lifecycle and post-approval planning for COVID-19 vaccines as well as future directions including international regulatory cooperation.EMA’s experience of the CMC scientific considerations, which facilitated both timely approvals (as Conditional Marketing Authorisations) and rapid increase in production capacity and supply, is of interest to healthcare professionals, academia, pharmaceutical industry and global regulators to communicate the flexibility and agility applied to COVID-19 vaccines by the EU regulatory system and how these activities can be optimised while complying with the strict quality standards in the EU.

Thinking big: the need for innovation in the production of lentivirus-based cell and gene therapies

<b>Elisa Manzotti</b>, Founder, BioInsights, speaks to <b>Hanna Leinonen, John Moscariello, Lee DaviesScott Cross </b>and <b>Will Junker</b> <div class="authors authors-lg"><div class="author"><div class="author-img"><img src="https://cdn.insights.bio/uploads/H Leinonen copy.jpg" data-image="1"></div><div class="author-det"><h3 class="author-name">Hanna Leinonen</h3><h5 class="author-title">Senior Scientist, Kuopio Center for Gene and Cell Therapy</h5> Hanna M. Leinonen holds a PhD in Molecular Medicine from the University of Eastern Finland (2014). She has 15 years’ experience on lentiviral vectors and 6 years’ experience on bioprocessing. She is currently working as a Senior Scientist in Kuopio Center for Gene and Cell Therapy (Finland) focusing on development of upstream processing for viral vectors (lentivirus, adenovirus, AAV). </div></div></div><div class="authors authors-lg"><div class="author"><div class="author-img"><img src="https://cdn.insights.bio/uploads/26118d9d52f34a12859025a3d5f4aabc.jpg" data-image="1"></div><div class="author-det"><h3 class="author-name">John Moscariello</h3><h5 class="author-title">Executive Director, Viral Vector and Gene Editing Process Development, Bristol Myers Squibb</h5> John Moscariello currently serves as Executive Director of Viral Vector and Gene Editing Process Development at Bristol-Myers Squib (BMS). At BMS, John’s group develops viral vector processes that enable rapid timelines to generate clinical viral vectors as well as the development and characterization of commercial viral vector processes. Prior to his work at BMS, John was the Vice President of Process Development at AGC Biologics where his team was responsible cell line development, upstream and downstream process development, analytical and formulation development, and technical support for AGC Biologic’s commercial manufacturing facility and supported development activities from generating processes for Toxicology/Phase 1 supply up to and including commercialization and post-approval process support. John is very active in the biotechnology community. He is on the Scientific Advisory Board for various conferences, including the BioProcess International conference series, and the CBI conference series on achieving efficient facilities and the next frontier of single-use technologies. John obtained his Ph.D. in Chemical and Biological Engineering from the University of Wisconsin-Madison, and his B.Eng. in Chemical Engineering from the University of Delaware. </div></div></div><div class="authors authors-lg"><div class="author"><div class="author-img"><img src="https://cdn.insights.bio/uploads/Lee Davies 1.jpg" data-image="1"></div><div class="author-det"><h3 class="author-name">Lee Davies</h3><h5 class="author-title">Director, Process Research &amp; Development, Oxford Biomedica</h5><p class="oneComWebmail-MsoNormal"><i>As Director of Process Research and Development, Lee helps drive development, innovation and improvements in&nbsp;</i><i>world-class lentiviral vector manufacturing processes,&nbsp;</i><i>applying scientific and engineering principles to all aspects of upstream and downstream process development, including process optimisation and scale-up, technology transfer to GMP manufacturing and process characterisation to support chemistry, manufacturing and controls (CMC) regulatory submissions.&nbsp;</i><i>As part of&nbsp;the</i><i>&nbsp;Process Research and Development team, Lee works</i><i>&nbsp;at the&nbsp;cutting edge of gene therapy, ensuring its applicability to the needs of the wider gene and cell therapy community. The team’s mission covers the&nbsp;</i><i>life cycle of gene therapies, taking</i><i>&nbsp;processes from the early phase development work through to&nbsp;fully optimised large scale, GMP-ready processes, manufacturing and commercialisation.&nbsp;</i><i>Prior to joining Oxford Biomedica 6 years ago, Lee worked at the University of Oxford as a postdoctoral researcher and obtained a D.Phil in Gene Therapy for Cystic Fibrosis.</i> </div></div></div><div class="authors authors-lg"><div class="author"><div class="author-img"><img src="https://cdn.insights.bio/uploads/Scott_122321_WebReady copy.jpg" data-image="1"></div><div class="author-det"><h3 class="author-name">Scott Cross</h3><h5 class="author-title">Senior Principal, Dark Horse Consulting</h5> Scott has over 20 years experience working with viral vectors, vaccines, and biologics in GMP environments. He has been responsible for cleanroom design, build out and commissioning of vector facilities, as well as the oversight of viral vector, cell therapy, fill finish and manufacturing support departments. Scott started his career at Merck and Co. where he worked on the development of an Adenovirus-based HIV vaccine and later, the development, optimization, and validation of release assays for live virus vaccines. He then moved to Indiana University (IU) where he managed the IU Vector Production Facility (IUVPF), overseeing GMP production and testing of Lentiviral and Retroviral vectors for the IUVPF and the National Gene Vector Laboratories (NGVL). While at the IUVPF he also managed the design, build and commissioning of a new viral vector GMP production and testing facility. After leaving the IUVPF, Scott joined Cincinnati Children’s Hospital Medical Center (CCHMC) managing the GMP Vector Production Facility, Viral Vector Core, and the Aseptic Processing Labs. Scott then moved to the University of Florida where he was the Director of Cell Therapy, Fill Finish and Manufacturing Support Operations for Florida Biologix and Brammer Bio. He was also responsible for the design of two new fill finish suites. Most recently, Scott served as Vice President of Vector Operations at Orchard Therapeutics where he was one of the original ten members and responsible for viral vector development, GMP vector production, plasmid production, oversight of 10 CDMOs, due diligence, and facility design. </div></div></div><div class="authors authors-lg"><div class="author"><div class="author-img"><img src="https://cdn.insights.bio/uploads/Will Junker headshot copy.jpg" data-image="1"></div><div class="author-det"><h3 class="author-name">Will Junker</h3><h5 class="author-title">Head of Vector Manufacturing Quality, Kite Pharma</h5> Will Junker is Head of Vector Quality and oversees the operation of QC, QA and QE to bring Kite’s vector manufacturing facility on line through licensure. Before this role, Will built Kite’s Quality Engineering function globally. His 35 year career in BioPharma includes 13 years at AstraZeneca and 15 years at Aventis/Sanofi as well as a few years at Baxter BioScience in technical/quality roles of increasing responsibility. </div></div></div>

Current Approaches for Dissolution Similarity Assessment, Requirements, and Global Expectations.

This report summarizes podium presentations and breakout sessions from the second day of the 2019 M-CERSI workshop on In Vitro Dissolution Similarity Assessment in Support of Drug Product Quality: What, How, and When? Presenters from the U.S. Food and Drug Administration (FDA), Health Canada (HC), European Medicines Agency (EMA), Brazilian Health Surveillance Agency (ANVISA), and the pharmaceutical industry shared experiences/concerns with dissolution profile similarity assessment supporting minor/moderate Chemistry, Manufacturing and Control (CMC) changes. Members from regulatory agencies explained that dissolution profile similarity testing is only part of the overall assessment of the acceptability of the proposed changes; decisions are usually made based on aggregate weight of evidence. Scientific shortcomings of f2 were highlighted but no proposal on how to replace it was made. Controlling dissolution timepoint variability and application of pairwise batch-to-batch comparisons (PBC) of dissolution profiles caused considerable debate. Several industry participants suggested increased sample sizes to raise confidence in decision-making and to avoid PBC. They proposed identification of a single mathematical method with predefined acceptance criteria and suggested that dissolution timepoint selection should follow EMA and HC guidance. A majority of meeting attendees favored applying clinically relevant dissolution specifications (CRDS) and dissolution safe space to determine the impact of minor/moderate CMC changes as opposed to dissolution profile similarity assessment via statistical methods. Day 2 of the workshop highlighted the need and opportunities for global harmonization including variability, timepoint selection, role of CRDS, and statistical methods to address the ambiguity globally operating pharmaceutical companies are currently facing.

Clinical Pharmacology Perspectives for Adoptive Cell Therapies in Oncology.

Adoptive cell therapies (ACTs) have shown transformative efficacy in oncology with five US Food and Drug Administration (FDA) approvals for chimeric antigen receptor (CAR) T-cell therapies in hematological malignancies, and promising activity for T cell receptor T-cell therapies in both liquid and solid tumors. Clinical pharmacology can play a pivotal role in optimizing ACTs, aided by modeling and simulation toolboxes and deep understanding of the underlying biological and immunological processes. Close collaboration and multilevel data integration across functions, including chemistry, manufacturing, and control, biomarkers, bioanalytical, and clinical science and safety teams will be critical to ACT development. As ACT is comprised of alive, polyfunctional, and heterogeneous immune cells, its overall physicochemical and pharmacological property is vastly different from other platforms/modalities, such as small molecule and protein therapeutics. In this review, we first describe the unique kinetics of T cells and the appropriate bioanalytical strategies to characterize cellular kinetics. We then assess the distinct aspects of clinical pharmacology for ACTs in comparison to traditional small molecule and protein therapeutics. Additionally, we provide a review for the five FDA-approved CAR T-cell therapies and summarize their properties, cellular kinetic characteristics, dose-exposure-response relationship, and potential baseline factors/variables in product, patient, and regimen that may affect the safety and efficacy. Finally, we probe into existing empirical and mechanistic quantitative techniques to understand how various modeling and simulation approaches can support clinical pharmacology strategy and propose key considerations to be incorporated and explored in future models.

The Chemistry Manufacturing and Controls (CMC) section of gene therapy-based INDs: overview in a changing landscape

There are many product development considerations for human gene therapy investigational new drug (IND) applications to ensure the proper ‘Chemistry, Manufacturing, and Controls’ (CMC) is achieved. Unlike small molecule drugs or biologics intended for repeated dosing, gene therapies are designed to modify patient cells directly or indirectly (administration of genetically modified cells to patients) to confer long-lasting therapeutic effects. The complexity of gene therapy manufacturing and therefore control of the processes is continually evolving with the need for disease specific innovations as well as standards across the various gene delivery platforms. As data becomes available on the durability, efficacy, and safety of gene therapy treatments post-administration, new factors must be considered when designing transgene cassettes, gene delivery vehicles, and conditioning treatments for infusion. Such clinical outcomes ultimately affect manufacturing and the continual evaluation of CMC requirements. This article highlights regulatory considerations and new guidance available for gene therapy manufacturing in a continually evolving landscape.

Challenges in AAV manufacturing: the interplay of process variations, in-process characterization, and drug product quality

A large focus for cell and gene therapy in recent years has been Chemistry Manufacturing and Controls (CMC) regulations related to production processes and analytical testing. Regulatory agencies have issued multiple guidelines for these novel biological products focusing on improving production, reproducibility, and testing expectations. With the increase in the number of clinical trials, there has also come a suite of clinical holds related to product quality or process reproducibility concerns. In remarks released from the FDA in 2018, they discussed how in contrast to traditional drug development, nearly 80% of the review process in cell and gene therapy applications is focused on the manufacturing process and product characterization methods. The focus of this article is discussing the current modalities for manufacturing, the available methods for evaluating in-process and final product characteristics, and the impact both the complex matrices and final formulation have on understanding changes to product quality and performance.

VectorMOD: Method for Bottom-Up Proteomic Characterization of rAAV Capsid Post-Translational Modifications and Vector Impurities.

Progress in recombinant AAV gene therapy product and process development has advanced our understanding of the basic biology of this critical delivery vector. The discovery of rAAV capsid post-translational modifications (PTMs) has spurred interest in the field for detailed rAAV-specific methods for vector lot characterization by mass spectrometry given the unique challenges presented by this viral macromolecular complex. Recent concerns regarding immunogenic responses to systemically administered rAAV at high doses has highlighted the need for investigators to catalog and track potentially immunogenic vector lot components including capsid PTMs and PTMs on host cell protein impurities. Here we present a simple step-by-step guide for academic rAAV laboratories and Chemistry, Manufacturing and Control (CMC) groups in industry to perform an in-house or outsourced bottom-up mass spectrometry workflow to characterize capsid PTMs and process impurities.