Overview of Nitrosamine Survey Findings and Nitrosamine Exchange
USP is working on several initiatives to address the emergence of nitrosamine impurities and related recalls in order to protect patients and strengthen the global medicines supply chain. USP aims to provide tools and resources to support industry and regulators to ensure safe drugs, as related to nitrosamine issues.
In this session, USP will be presenting key findings from the nitrosamine survey conducted recently to understand current challenges faced by our stakeholders in testing and controlling nitrosamine impurities; not just in drug substance and drug products but also in excipients. We wanted to also gain insights about what else is required in this space by industry stakeholders and regulators across the globe.
USP will also provide a brief overview of Nitrosamine Exchange – a community hub launched this year with a purpose to exchange nitrosamine knowledge and share ideas with other members in the hub. USP’s “Nitrosamine Exchange” will be a place to learn and share best approaches to implement nitrosamine risk assessments.
Speaker: Naiffer Romero, USP
Bioanalytical Considerations for Adeno-Associated Virus (AAV) Gene Therapy Drugs
Viral vector-based gene therapies continue to grow with several products now approved and a flowing pipeline of candidates seeking approval in the coming years. To date, the most commonly used vectors are adeno-associated virus (AAV) based. The FDA does not currently provide guidance on how to approach immunogenicity assay development and validation when using emerging or existing technologies - a requirement to determine exposure in the preclinical and clinical phases of drug development. As a result, there is an increased need for scientific expertise to develop bioanalytical strategies utilizing new technologies as well as existing platforms within the regulated bioanalysis space to assess the safety of new AAV products. In this eChalk Talk, we will discuss considerations for AAV-based gene therapy programs, and best approaches and strategies for development and validation of assays to support regulated bioanalysis.
Speaker: Brad Reinholt, CRL
Unlock the Full Potential of Your Lab: CrossLab Asset Monitoring
Today’s labs are large and complicated with complex equipment, numerous vendors, IT infrastructure security requirements, difficult data collection methods and many other challenges. If you are responsible for a laboratory, you are keenly aware of the effort it takes to gather data to make decisions with confidence targeted to improve results. It is difficult to get full visibility because of geographical differences, new acquisitions, data collection methods, analysis and a variety of other reasons.
Learn how to use lab-wide instrument data to make capital expense decisions, reduce operational expenses, and improve productivity. Understand the hidden costs of not collecting instrument usage data. Show internet-of-things (IoT) technology available today to help lab managers measure instrument usage.
Speaker: Melissa Zeier, Agilent
Accurate Determination of Mutagenic Azido, NDSRI and Nitrosamine Impurities in Drug Substances and Products Using LC-MS/MS
Several sartan, ranitidine and metformin-based drug products were recalled due to the nitrosamine contamination in the respective drugs products and drug substances in unacceptable amounts. Since then, pharmaceutical companies are under increased scrutiny by the global regulators to accurately identify and quantify mutagenic impurities in drug substances and products. The USFDA and other regulatory agencies have published several analytical testing guidance’s to evaluate mutagenic nitrosamine impurities in these drug substances and drug products. Recently, sartan therapeutics were again recalled by various regulatory agencies due to the presence of another known mutagenic impurity called Azido Biphenyl Tetrazole (AZBT) impurity. Additionally, there were recalls related to nitroso impurity in Varenicline drug products. In this presentation, we will cover the accurate determination of mutagenic azido impurities, NDSRI (Nitroso Drug Substance Related Impurity) and nitrosamine impurities in drug substances and drug products using LC-MS/MS based analytical strategies for the confident identification and quantification approaches that you can adopt to meet your nitrosamine detection and quantitation needs.
Speaker: Kartheek Srinivas Chidella, M.Sc. , Agilent
Application of Expert System with Augmented Intelligence in Regulated Bioanalysis
One of the biggest challenges bioanalytical scientists’ faces is in generating quality data while ensuring compliance to a complex set of compliance requirements in a timely manner. In most cases, when scientists discover an issue, it is too late, and it is always desired to prevent issues before it can happen. Over the last decade, tools like Lean and Six Sigma improved the processes involved in regulatory bioanalysis, in most cases, these tools fail to anticipate and prevent failure from happening.
An expert system with augmented intelligence can automate the manual, error-prone and time intensive process of data entry, analysis and review. The system can alert users of potential issues, in some cases before it happens. Here we present three case studies to demonstrate how an expert system can help anticipate issues and prevent it from happening. This can result in significant improvement in quality of the data and saving time and money.
Mohammed Moin, Ph.D., Somru
Clarinda Islam, Ph.D., Somru
How Oncology Drug Developers Can Prepare for FDA’s Project Optimus
Historically, the dosing strategy for oncology drugs has focused on use of the maximum tolerated dose. This has resulted in drugs’ pharmacokinetic (PK) profiles, pharmacokinetic/pharmacodynamic (PK/PD) relationships, and clinical target inhibition largely being ignored in the dose decision making process. Thus, cancer patients often struggle to tolerate their medication long-term, requiring dose modifications including dose reductions and holidays. What’s more, for many oncology drugs, their dosing or schedules have been modified to address safety or tolerability issues after regulatory approval.
These challenges spurred the FDA’s Oncology Center of Excellence to develop a new initiative called “Project Optimus” to addresses issues relating to dose optimization in clinical trials assessing the safety and efficacy of oncology drugs.
What does this shift in dose optimization during drug development mean for the small biotechs that develop more than 80% of novel compounds? And what steps should they take regarding their upcoming clinical trials?
Speaker: Julie Bullock, Ph.D., Certara
Meeting Compliance Requirements with UV-VIS Systems
UV-VIS measurements are an integral part of routine work in today’s labs. Whether implemented as a standalone technique or as an accompanying measurement to other processes, a UV-VIS spectrometer can be utilized in production processes to provide measurements for both development and quality control. An absorption spectrum can be used for both qualitative and quantitative verification. This is done through assigning absorption peaks to a particular molecular species and determining values for sample concentration and melting temperature.
Meeting compliance standards in order to bring a UV-VIS spectrometer into production is a necessary aspect of ensuring quality in a regulated pharmaceutical environment. The basic goals of doing this are to provide documentation of the “who, what, where, when, and why” for each measurement. In addition, verification tests are necessary for compliance to confirm the accuracy of the spectrometer and therefore the validity of the measurements that are performed.
In this talk, we will discuss a basic overview of meeting compliance requirements for operating a UV-VIS spectrometer and how the vendor’s software package for the spectrometer can streamline these requirements.
Speaker: Scott Melis, Ph.D., Agilent
Novel Hepatocyte Suspension-Based Assay Formats for BSEP and MDR3
Drug-induced liver injury (DILI) accounts for >50% acute liver failures, and is the leading cause of drug development failure, and boxed warning and market withdrawal of approved drugs. Inhibition of BSEP and MDR3 is one of the underlying mechanisms for DILI. BSEP and MDR3 are the primary hepatic transporters responsible for exporting bile salts and phosphatidylcholine, respectively. In humans, dysfunction of BSEP and/or MDR3 can result in serious liver injury. The discussion will focus on the patented assay platforms BSEPcyte® and MDR3cyte®, which are developed to determine the inhibition of BSEP and MDR3 activity using primary hepatocytes in suspension from human and preclinical animal species, respectively. The assay formats are physiologically relevant, accurate, and flexible for cross species comparison.
Speaker Kan He, Frontage/Biotranex
API Sparing Dissolution Rate Determinations for Discovery
Allied to solubility dissolution rate is an important measure of a lead’s developability, we have previously presented on small scale form dependent solubility measurements, in this presentation we will focus on material sparing dissolution rate assays. The original and still used Wood’s Apparatus for Dissolution Rate measurement requires grams of material, whereas more recent miniaturized technologies have reduced the material consumption to the sub gram scale.
Nevertheless, the sample consumption of these technologies is still a limiting factor for acquiring critical Dissolution Rate information for lead series compounds with the amounts of available material in early drug development. This contributes to a a mismatch between AI driven models and supporting those models with real data.
We will discuss determining high-quality Dissolution Rate from only a fraction of unprocessed solid material. Less than 100 micrograms per buffer condition. One milligram of sample enables 10-20 separate assays whilst still considering the solid form of the starting material.
Discussion of reliable Dissolution Rate comparable to traditional methods, readily measured from unprocessed powder samples below 100 µg using the image-based single particle analysis SPA™method. Earlier data acquisition accelerates drug development by providing data for solid form (polymorph , salt, co-crystal) and particle size selection as well as PK models and formulation development.
Speaker: Sami Svanbäck, Ph.D. , The Solubility Company
On the Mark: Identifying Non-Invasive Biomarkers of Target Engagement
A key aspect of the drug discovery pipeline is evaluation of in vivo target engagement and subsequent biological response. While measures of target engagement are often optimized in cellular and preclinical systems, direct measures of tissue target engagement are often limited in human subjects as part of early clinical studies. Circulating small molecule biomarkers in blood may serve as a robust means for measure of target engagement, providing a non-invasive approach for understanding drug dosing, tissue penetration, target engagement, and therapeutic response. In this session, we will discuss methods for the rapid discovery of small molecule biomarkers of target engagement, leveraging integration of next generation mass spectrometry, genomics, and experimental systems.
Speaker: Mohit Jain, Sapient
Monkeypox: What is it, What therapies exist, and What Research Tools Exist to Study and Evaluate Interventions
Recent outbreaks of Monkeypox have brought the long-studied virus into the popular press. There is concern over the findings of cases in areas that have not previously shown diagnosed infections regionally, and concern over potential opportunities for treatments that may exist. Monkeypox is an othopox virus that has been studied for many decades both for concern of communicable disease but also concern it could be used as a warfare agent. This e-chalk talk will provide information on what the virus is, how it is transmitted, and what current treatments are available. Since Lovelace helped perform studies that led to the approval of the current antiviral for monkeypox (ST246), we will present on the studies that are performed to develop new (and old) treatments. The characteristics of the studies that are performed to prove the drugs work and are safe will be described, using our previous experience as an example.
Speaker: David Revelli, Ph.D., Lovelace Biomedical
Mapping the Missing Link- Bridging the Gap Between Human Genetics and Disease Biology Through Small Molecule Biomarker Discovery
Genome-wide association studies (GWAS) have been transformative in elucidating the genetic landscape of human disease, defining hundreds of common and rare genetic variants that contribute to the development of disease. The challenge to date, however, lies in understanding the biological mechanisms that underlie these genetic associations and identification of key targets for therapeutic intervention. Circulating small molecule biomarkers provide an opportunity to bridge the gap between large-scale genetics data and biological understanding.
In this session, we will discuss approaches for the comprehensive assay of small molecule biomarkers on a population-scale and application of these biomarkers to elucidate mechanisms linking genetic variants and human disease.
Speaker: Mohit Jain, Sapient
Answering Frequently Asked Questions Around Challenges When Using qPCR, ddPCR and NGS in Regulated Studies for Cell and Gene Therapy Products
During our webinar in April 2022, we discussed the role of molecular testing platforms, such as qPCR, ddPCR and NGS in enabling discovery and development of cell and gene therapy products. While these technologies are well-developed and well-supported in other areas of drug discovery and development, their presence in bioanalysis of cell and gene therapy products is a more recent development. This has resulted in many unanswered questions on the appropriate use of these technologies in the field of bioanalysis, especially due to the lack of prescriptive guidance. We will use a redacted and anonymized qPCR case study to answer some of the questions that frequently come up. We will then open the floor to discuss approaches to some common questions and requests.
Speaker: Stephanie Farmer, Ariadne Solutions
New Software Tools for Impurity Analysis of Synthetic Oligonucleotides
Oligonucleotide therapeutics have emerged in recent years as a powerful alternative to small molecule and protein therapeutics. Manufacturing and quality control of oligonucleotide therapeutics requires highly selective and sensitive LC-MS methods. This presentation will highlight two recently introduced software applications: one used for purity analysis and rapid intact mass confirmation of oligonucleotide impurities, the second one for fast data interpretation of complex oligonucleotide fragmentation spectra. These two applications are embedded in an automated, compliance-ready LC-MS workflow designed for analysis of synthetic oligonucleotides and their impurities.
Speaker: Catalin Doneanu, Ph.D., Waters
Increasing the Productivity of Oligonucleotide Purification through Column Scaling and Method Optimization
Isolation of purified oligonucleotides in an efficient, cost-effective manner can be extremely challenging. Reverse-Phase UPLC using an ion-pairing system of HFIP-TEA has become the gold standard for oligonucleotide analysis. This column and mobile system capitalize on the benefits of small columns particles, along with the unique ion-pairing properties using HFIP. The result is a rapid efficient method to accurately assess oligonucleotide’s purity.
The process of scaling an oligonucleotide separation using a UPLC method with a HFIP-TEA ion pairing buffer to preparative scale using a 3 cm ID column will be discussed. The components of each step of the process will be systematically evaluated to understand the impact of its variation and its impact on the overall process. The different items tested and varied include mobile buffers, mass and volume column loading and fraction collection.
Speaker: Paul Lefebvre, Averica Discovery Services (Waters)
Avoid getting "lost in translation" - Increase confidence in translational research using biosimulation
One of the pivotal milestones of early drug development is obtaining approval for an investigational new drug application (IND). A proposed first-in-human (FIH) study design is required for every IND application, of which a robust FIH dose rationale is a critical component. Physiologically-based pharmacokinetic (PBPK) modeling is a methodology based upon in-depth mechanistic understanding of the biological and pharmacologic phenomena which determine in vivo PK in animals and humans. Consequently, PBPK models are well-posed to make robust predictions of in vivo PK exposures, even when based on the limited data available during the initial stages of drug development. As such, PBPK approaches are being increasingly employed to strengthen the FIH dose rationale.
Other applications of PBPK in early development include guiding early formulation development and facilitating lead candidate selection. This eChalkTalk will focus on how PBPK modeling can be applied during early drug development with a particular focus on optimal FIH candidate and dose selection, thereby enhancing the odds of a successful IND and subsequent clinical development.
Speaker: Mike Lovern, Certara
Cytokine Multiplex Analysis in Non-Clinical Studies
This eChalk talk will explore the possibilities and challenges in the analysis and interpretation of multiplex cytokine data in the context of non-clinical safety studies. The focus of the discussion will be on strategies to improve the application of multiplex analysis to obtain a better understanding of adverse immune events or immunotoxicity in non-clinical models.
Speaker: John Farmer, Lovelace
Overview of pharma outsourcing for CRO/CDMO
Overview of pharma outsourcing services provided by contract research, development, and manufacturing (CRO-CDMO) organizations for small molecules and biologics businesses.
We will talk about the value addition that CROs and CDMOs bring into the drug development lifecycle in today’s fast-changing world with scalable and sustainable solutions. The CRO/CDMO market is currently growing globally at a tremendous pace with India and China at the center of it. While the CROs and CDMOs are increasingly becoming an integral part of the pharma and biotech industry, it is also important to know how Syngene has transformed its supply chain to ensure business continuity amidst disruptions, enable seamless operations for the business and extract value from its supplier ecosystem.
Speaker: Veera Oruganti, Syngene
ELISpot Assay Assessments and Method Validation for Next Generation Therapeutics
This presentation will review the current state of the ELISpot assessment and the validation approaches for test methods using ELISpot assessment as analytical methodologies and discuss the extent of validation requirements based on the intended use of the assays.
The enzyme-linked immunosorbent spot (ELISpot) assay is a well-established and highly sensitive quantitative assay used to measure the secretion of proteins such as cytokines and growth factors. Regulatory guidance documents are not available for the validation of ELISpot assays despite being used since the early 1980’s, primarily in the field of vaccines. While comprehensive bioanalytical guidance documents released by the regulatory agencies focus on methods for nonclinical and clinical pharmacokinetic and toxicokinetic studies for small and large molecules, many of the standardized requirements for method development and validation are difficult to apply to ELISpot assays due to the unique features of those assays. Organizations, including the Workshop on Recent Issues in Bioanalysis (WRIB), have been working to harmonize divergent global practices for method development and validation. However, the approach in conducting those practices vary from one laboratory to another based on scientific interpretation of the published recommendations. Historically, the ELISpot was mainly used to measure T cell responses through the secretion of cytokines in the context of vaccine development. With the rapid growing field of new therapeutics, such as cell and gene therapies, use of the ELISpot assay to assess the safety of some of these therapeutics has become more common.
Speaker: Paolo Campoli, CRL