Agnes Shanley is senior editor of Pharmaceutical Technology.

Agnes Shanley

For the past two decades, regulators have voiced support for the development of a more flexible, agile approach to developing and manufacturing pharmaceuticals (1). Miniaturized, modular, and continuous manufacturing processes will help speed scale-up, address supply chain complexity, and prevent shortages of important medications, Janet Woodcock, director of FDA’s Center for Drug Evaluation and Research (CDER), noted during a meeting of the US National Academy of Sciences (NAS) in late February 2020. The COVID-19 pandemic, which was then at its earliest stages in the United States, has exposed vulnerabilities and emphasized the need for alternatives, she said (2).

Different approaches are being refined to address these challenges. One area of growing interest is three-dimensional printing (3DP), which allows greater customization of dosage forms, including polydrugs that would incorporate dosages of multiple active ingredients in a single form to enhance patient compliance and overall outcomes. The technology gained a foothold in pharma when FDA approved Spritam, an anticonvulsant manufactured by Aprecia Pharmaceuticals using 3DP technology.

Aprecia offers its 3DP platform for license to other companies. Its technology, ZipDose, uses powder liquid binder jetting to create a wide range of rapidly dissolving dosage forms and other solutions, says CEO Chris Gilmore. The process has been successfully applied to a wide range of drugs ranging in solubilities from Biopharmaceutical Classification System (BCS) Class I to IV, and incorporating a variety of engineered particle technologies, he says.

In 2020, Aprecia formalized a joint collaboration with Purdue University to advance 3DP in pharma. Eric Munson, head of Purdue’s School of Industrial and Physical Pharmacy, expects the collaboration to help drive the formulation and manufacturing of new drugs with properties that are not always feasible using traditional approaches (e.g., rapid dissolution, high drug loadings, and flexible manufacturing scales). Purdue will contribute knowhow to the venture, both in equipment design and drug-delivery science, he says.

Munson sees 3DP helping industry develop clinical trial supplies and create patient-centric dosage forms. “Scalability is potentially a huge step for manufacturing small-scale doses, which can then be easily adapted to large-scale manufacturing,” he says.

Aprecia currently out-licenses its platform to a number of pharmaceutical companies for lifecycle management, line extensions, and new chemical entities, says Gilmore, who envisions that any intellectual property created by the collaboration with Purdue would also also be outlicensed.

Different types of 3DP technologies, such as binder jetting and fused deposition modeling (FDM), may have different applications, Munson explains. “3DP in the pharmacy may involve more FDM-based approaches to avoid powders,”he says, “but also pose challenges with ensuring product quality, such as optimizing dissolution rates and stability. Binder jetting technologies, meanwhile, may allow for high-dose products with fast dissolution rates, and also enable appropriate taste masking of the particles prior to printing,” he says.

Recently, Gilmore says, the company has developed a scored, oro-dispersable tablet medication in excess of 1000 mg that rapidly disintegrates with a very small amount of liquid. Aprecia has also made advancements in modified release and poor solubility products, he says. “At this point, the combination of high-dose and fast-melt capabilities can only be achieved with 3DP, but the platform has the flexibility and precision to do more,” he says.

Other companies are also developing 3DP applications in pharma. In the UK, FabRx, a spinoff of University College, London, is actively working in this area.

Meanwhile, pharma continues to develop more continuous processing routes, mainly in oral solid dosage form (OSD) manufacturing, to improve scalability and efficiency. At February’s NAS meeting (2), Daniel Blackwood, a research fellow at Pfizer’s small molecules drug product design group, discussed the evolution of these efforts at Pfizer.

One driving force has been process intensification, he said, which enables miniaturization, in turn allowing for equipment modularization and portability. He envisions a time when, using miniature, modular equipment, pharma companies might routinely share space and even some manufacturing operations, without violating precompetitive agreements.

Research is also underway to develop miniature manufacturing lines, to optimize flexibility in response to market demands. In 2016, researchers at the Massachusetts Institute of Technology (MIT) developed a prototype line that could allow companies to manufacture specific OSD forms (3). 

Various R&D projects are carrying this work forward in different applications. For example, Govind Rao, a professor of chemical and biochemical engineering at the University of Maryland, is working on a portable system that would make FDA-approved biologics at the point of administration in less than 24 hours. The US Defense Advanced Research Projects Agency (DARPA) is funding this work, which would use freeze-dried formulations, adding buffer and DNA to activate them, synthesize RNA, and manufacture protein. Describing the research at the NAS meeting (2), Rao said his team has conducted a demonstration project involving Neupogen (filgrastim)and is working with GE on development of remote bioreactor sensors to monitor analytes, oxygen, and carbon dioxide in small-scale systems in real time (2).

Also funded by DARPA, research underway at MIT is developing miniaturized, automated equipment to speed up chemical synthesis (4).The project began with the goal of harnessing machine language to predict reaction pathways based on published results from the literature, says project co-leader, MIT Professor Klavs Jensen. These efforts continue in research underway with the Machine Language for Pharmaceutical Design and Synthesis, an academic-industry consortium, he says. 

Another aspect of this research, however, has been developing an integrated robotic system that would allow for the automation of synthesis reactions (4). The set up uses artificial intelligence (AI) to find the best possible pathways, allowing users to develop recipes for specific reactions. These recipes are then sent to a 2-m³ modular processing unit that features miniature mixers and other process equipment and a robotic arm that automates molecule production. Initial research evaluated results for 15 workhorse APIs. At the time, researchers noted a need for better predictive models for flow chemistry and purification, both of which remains a challenge today, Jensen says. However, since research results were first published, he notes, robotics have been improved, and now include integrated online measurement systems using infrared (IR), Fourier Transform IR, and high-pressure liquid chromatography and mass spectrometry so that key parameters can be measured in real-time, permitting online optimization.

OSD equipment processing manufacturers are working to improve continuous feeding (5,6). Pfizer has already made significant progress in developing continuous direct compression for OSD products. The company is also focusing on increased use of analytics, incorporating sensors to monitor mixing and allowing process control to enable real-time decision making, Blackwood said at the NAS event, noting that computational process modeling and data analytics are key focus areas (2).

Use of process analytical technologies (PAT) has become standard in many pharma development programs, and technology vendors have facilitated efforts by incorporating analytics into more process equipment. For the contract development and manufacturing organization (CDMO), Lonza Pharma & Biotech, PAT is finding increased use, along with more at- and in-line testing, to ensure process compliance, robustness, and faster product release, says Charles Christy, head of commercial solutions for Lonza’s Ibex Dedicate division. Use of Raman and nuclear magnetic resonance (NMR) spectroscopy, refractive index, excipient identification, metabolite monitoring, and cell counts has now become routine, in both process development and manufacturing, he says.

In mammalian cell-culture, Christy says, screening and optimization tools are finding increased use, both in clonal cell line development (e.g., microfluidic technology, such as Berkeley Light’s Beacon system, which screens and selects the best producing clonal cell lines) and in cell culture (e.g., using Sartorius Biotech’s Ambr systems), to speed time to clinic for high yield, robust processes.Single-use technologies are playing a more important role in both small molecules and biologics, Christy notes.Their combination with closed process systems and ballroom processing concepts has allowed facilities to be configured for concurrent manufacturing. In antibody production, for example, they allow the company to configure processes from standard to next-generation formats such as bi-specifics, or to process high titer or other challenging processes, Christy says.

In small molecules, single-use approaches are also proving important in highly active pharmaceutical ingredient (HAPI) manufacturing to improve operator safety and reduce toxic waste streams that would have been generated by traditional clean-in-place (CIP) activities, he says. Christy sees a growing trend to automate processes and expand use of analytics, which is improving efficiency, and reducing errors, manual intervention, and staffing requirements (i.e., overall full-time equivalent numbers). Capsugel, for example, is using AI for online testing, real-time quality assurance, and product release of capsule manufacturing, he says.

As the emphasis on scalability intensifies, technology vendors are collaborating more closely with CDMOs and also expanding their own contract services, with some offering turnkey CDMO services as well as processes and equipment designed to cut down on development time. For example, in OSD development, Adare Pharmaceuticals (formerly Aptalis), which acquired Orbis Biosciences in May 2020, offers platforms that include Microcaps for taste-masking; Diffucaps and Multi Mini Tablets, to control the rate of drug release; and Precision Particle Fabrication technology to control particle size distribution.

On the filling and packaging side, Syntegon (formerly Bosch) Packaging Technology has introduced products that include modular filling systems; microbatch systems for aseptic and HAPI applications; and visual inspection platforms that utilize AI. 

The company’s single object data acquisition system, offered in collaboration with Schott Smart Packaging, aims to help users improve product traceability and supply-chain transparency. The company has also expanded its range of pharmaceutical services, including programs that focus more closely on small-to-mid-scale operations.

1. FDA, "Pharmaceutical Quality for the 21st Century A Risk-Based Approach Progress Report," 

2. US National Academy of Sciences, “Innovations in Pharmaceutical Manufacturing Proceedings of a Workshop—in Brief,” 

3. J. Markarian, “Solid Dose Continuous Manufacturing Presses on,” 

5. A. Trafton, “Pharmacy on Demand,” 


Vol. 44, No. 10
October 2020
Pages: 34-37

When citing this article, please refer to it as: A. Shanley, "Improving Agility Through Scalability," 

Articles

Research is striving to make pharmaceutical processes more scalable by making them simpler and easier to replicate and control. Advances include 3D printing, as well as miniaturized and continuous processes, all of which are being aided by improved automation and analytics. 

Improving Agility Through Scalability

Over the past few years, the “4.0” suffix has become widely used (and, some might argue, hyped) in the pharmaceutical industry and beyond. Reflecting the extension of “Manufacturing 4.0” principles to biopharm, Biopharma 4.0 (1) centers around the use of sensors, process analytics, and real-time data. This approach allows integrated networks comprising the Industrial Internet of Things (IIOT) to capture, store, and analyze information from separate sources, and present it in optimal form. The result is improved cross-functional collaboration, predictive maintenance, and efficiency.

The operating systems required already exist. Combined with use of cloud computing, artificial intelligence, and technologies such as virtual and augmented reality, the principles behind BioPharma 4.0 are expected to decrease scale-up costs and timelines, improve manufacturing efficiency and quality control, and allow more effective models to be built for predictive maintenance and training.

Some manufacturers are piloting 4.0 approaches that are already yielding benefits for both small-molecule and biopharmaceutical operations (2,3). Others are still grappling with “3.0” or even “2.0” challenges and integrating the digital silos and islands of automation that remain within their operations.

For the biopharmaceutical industry, whose processes are more complex than those for most small-molecule drugs, adopting Biopharma 4.0 principles can help manufacturers address “formidable” challenges, says Mark

Demesmaeker, head of data analytics at Sartorius Stedim Biotech. In 2020, Demesmaeker notes, biologics will account for more than 25% of the pharmaceutical market, at a time when smaller volumes, higher product turnover, demands for lower cost of goods sold (COGS), improved product quality, and reduced time to market or clinic have become the norm. Factoring in the specialized requirements for personalized medicines, such as gene and cell therapies, and the introduction of more biosimilars, he says, and a more data- and analytics-driven approach is fast becoming “a cornerstone for future success” in biopharma development and manufacturing.

While investing in better automation and analytics, manufacturers can take advantage of modular, pre-validated, and pre-tested single-use equipment, bioreactors, and chromatography systems that include built in electronic batch record (EBR) and manufacturing execution system (MES) functionality. They can also utilize off-the-shelf modeling programs and improvements in single-use equipment automation.

Partnerships or acquisitions involving equipment vendors and process-control and software suppliers are helping to drive change. Today, all process-equipment vendors have allied with leading automation and control companies. For instance, Cytiva (formerly GE Healthcare Life Sciences) has connected its FlexFactory single-use platform to a Figurate distributed control system (DCS) since 2015. The company has formed partnerships with Rockwell, Emerson, and Siemens to bring plug-and-play automation to ppstream bioreactors and downstream chromatography scaleup for companies that might not be able to invest in foundational MES or EBR software on an enterprise level.

Recently, some of the greatest advancements have been made in automation for single-use technology, says Kevin Seaver, executive general manager of Cytiva’s automation and digital division. He recalls the early days of single-use equipment development at Xcellerex and working with sensor vendors and tubing welders on solutions. Today, the company can provide Figurate products that have already been tested and validated, to allow systems to work better together, enabling shorter implementation time for customers, he adds. “We have made sure that Figurate products are connected to platforms such as Rockwell PlantPAx, Emerson’s Delta V and Siemens PCS7,” says Joe Makowiecki, enterprise solutions director of business development at Cytiva. “Using automated FlexFactory approaches allows users to incorporate EBR or scheduling software. This reduces cost, provides an audit trail and releases batches faster. It also lets them use not only manufacturing but materials data,” he adds.

Another important change has been the move to different types of modeling, which has become vendor agnostic. “This process is become more ‘off the shelf’ with the goal of making it easier for end users to scale up without having to run extensive designs of experiments,” says Seaver.

In this same spirit, Sartorius introduced the third generation of its BIOSTAT STR single-use bioreactor, which runs the new Biobrain automation platform, designed to enable quick and easy system configuration. The system features integrated and redundant single-use sensors that provide real-time data to measure and control critical process parameters such as pH, dissolved oxygen, viable biomass, glucose, lactate, and foam.

“Non-invasive measurements save set-up time, prevent interface issues and reduce the need for off-line manual sampling to enhance batch-to-batch consistency,” says global consultant Kevin McHugh. Users can incorporate feed controls, bleed controls, and connectivity to Repligen’s XCell ATF cell-retention devices to increase cell density and productivity, he says. Connection to Umetrics SIMCA, meanwhile, allows them to harness multivariate modeling and control, enabling them to predict yield and optimal harvest time points at the early stages of the perfusion process. The system can be directly integrated locally via a control tower or with a data historian, or more broadly via DCS platforms from Emerson, Siemens, or Rockwell.

As they explore off-the-shelf solutions, more companies are also investing in improved analytics and process control and investigating more of the building blocks of Biopharma 4.0. In scale-up, process intensification is driving the use of more advanced analytics. “About 30% of new molecules moving into clinical manufacturing will be implementing intensified processing in upstream or downstream operations,” says Andrew Bulpin, head of process solutions at MilliporeSigma. As a result, most companies realize that they need to upgrade their process analytics and automation. “Today’s processes already produce in excess of 1 billion data points per run, and with the adoption of intensified processing technologies with advanced sensor technologies and enhanced data capture capabilities, the volume of data and information gathered per batch is exploding,” Bulpin notes. Advanced analytical tools are required to ingest, normalize, analyze, and enable decision making at high volume and speed, he says.

Currently, every key industry organization, from the International Society for Pharmaceutical Engineers (ISPE) to BioPhorum, has a Biopharma 4.0 initiative underway. January 2020 marked the official start of the Biopharma 4.0 Alliance, a partnership between the National Institute for Biotechnology Research and Training (NIBRT) in Ireland and the Boston Consulting Group.

The alliance is working with 17 different software, control, and equipment vendors as well as biopharmaceutical manufacturers on real-world Biopharma 4.0 use cases. A demonstration center (Photo) is now up and running to introduce more industry professionals to the concepts involved.

A number of biopharma equipment vendors have introduced products that directly apply Biopharma 4.0 concepts. Sartorius Data Analytics, for instance, is launching a modern data-science platform for process development and data management, and to support the trend to open-source data modeling (e.g., using the Python coding language) and hybrid modeling, which blends statistical and kinetic and metabolic modeling techniques, says Demesmaeker.

MilliporeSigma has launched the Bio4C Software Suite, which combines process control, analytics, and plant-level automation, allowing users to visualize, analyze, and control the entire manufacturing process and individual operational units, enabling enhanced process control and insights, says Bulpin. ProcessPad, another recent introduction, is designed to help users acquire, aggregate, and analyze data from equipment, paper and electronic records, and facility databases, to automate the generation of continuous process verification reports.

On the sensor level, MilliporeSigma introduced Raman spectroscopy sensors and accompanying software to provide real-time process analytics across multiple critical process parameters via in-line measurement. Sartorius also rolled out the BioPAT Spectro sensor platform, which allows users to integrate Raman spectroscopy into ambr 15 and 250 development platforms as well as Biostat STR development, pilot, and production systems, says McHugh.

In reactor modeling, McHugh says, techniques such as tip-speed scaling are being replaced by more advanced and holistic approaches that weight many factors at the same time (e.g., power-per-volume, gas transfer, carbon dioxide stripping, eddy size, shear stress, and volume effects) for the entire duration of the process.


Vol. 44, No. 8
August 2020
Pages: 27–28

When referring to this article, please cite it as A. Shanley, “Skipping Steps to BioPharma 4.0,” 

Even though many companies are at early stages of digital automation, more are evaluating BioPharma 4.0 tools and leveraging built-in automation to speed scale-up and boost efficiency. 

Skipping Steps to BioPharma 4.0 

Nitrosamines are a group of genotoxic compounds that have been studied widely in the food, water treatment, and chemical industries. In fact, the compound N-Nitrosodimethylamine (NDMA, also known as dimethylnitrosoamine) may be one of the most closely studied carcinogens in the world, noted David Light, CEO of Valisure, an independent analytical pharmacy, who spoke on a webcast on nitrosamine contamination on July 14 (1). First identified as toxic in 1956, NMDA was the topic of a US Senate panel investigation in 1977, and a World Health Organization and United Nations Summit in 1978, Light told attendees at the webinar. Analytical methods have been available to detect the compound and other nitrosamines for over 50 years, he noted.

Yet the pharmaceutical industry was blindsided in 2018, when trace levels of NDMA were found in finished blood-pressure and heart medications, the ARBs (angiotensin receptor blockers) valsartan and losartan and a number of their APIs (2). The problem led to a cascade of product recalls around the world and confusion, as healthcare providers and patients sought substitutes.

This particular set of recent contamination events was traced to a number of problems. One API manufacturer had optimized its manufacturing process yet failed to detect potential chemical synthesis problems that could lead to generation of contamination. In other cases, manufacturers had been using recycled solvents and other materials but failed to work through the potential manufacturing risks.

In September 2019, for very different reasons, NDMA was found in ranitidine and the name-brand drug Zantac. As Ramin (Ron) Najafi, CEO of Emery Pharma, noted during the webinar, ranitidine products had been on the market since the 1980s, and in 2017 ranitidine was the 48th most widely prescribed medication in the United States.

In this case, extensive research verified that the NDMA was not a process impurity, but the result of the ranitidine molecule’s inherent instability, so that it formed NDMA when exposed to high temperatures and humidity. Emery and Valisure both filed citizen’s petitions with FDA (3,4), and Emery developed an ultra high-pressure liquid chromatography-mass spectrometry (UHPLC-MS/MS) method to quantify NDMA down to 1 ng/tablet. Eventually the drug was taken off the market. However, in late 2019 and early 2020, NDMA contamination was found in yet another high-volume pharmaceutical, metformin, used to treat diabetes and a number of other illnesses. Valisure filed a citizen’s petition with FDA to draw attention to this problem in March 2020 (5).

As these events suggest, gaps remain in the industry’s knowledge regarding genotoxic compounds and how they may form in pharmaceutical processes, and how to prevent their presence in pharmaceutical products.

An informal poll of the audience at the July webcast, even if statistically irrelevant, sketched the framework for potential problems. When asked whether their companies had formal quality control (QC) testing procedures in place to test for NDMA and nitrosamines, 32 professionals responded, with nearly 41% saying that their companies had no QC procedures in place identify nitrosamines in their products and process streams. Approximately 16% said that they routinely test all products for nitrosamines, 24% that they test for nitrosamines in some products, and 19% that they only test for products that are in developmental, rather than commercial stages.

Although the agency’s scientists continue to study the problem, FDA has not issued any new guidance on nitrosamines since 2017, although FDA supports the use of the International Council for Harmonization’s (ICH’s) M-7 guidance, a key reference for many in the industry (6).

During the July webcast, Ed Gump, vice president for small molecules with the United States Pharmacopeial Convention (USP), emphasized the need for risk-based approaches to preventing genotoxic contamination. As he noted, the ICH M7 guidance requires identification, categorization, qualification and control of DNA reactive mutagenic impurities to limit carcinogenic risk. A risk level of 1 in 100,000 is currently considered negligible, he said, but where a potential risk has been found for an impurity, manufacturers must develop an appropriate control strategy that leverages process understanding. In addition, analytical controls should be developed to ensure that any mutagenic impurities are present at or below the acceptable cancer risk level.

To this end, Gump suggested, manufacturers of all types must prioritize the evaluation and testing of specific drugs, drug classes, and manufacturing materials that are more likely to contain unsafe levels of genotoxic impurities, using predictive and risk-based approaches. In addition, he noted the need to use an analytical method that has been widely demonstrated to be fit for its intended purpose for testing impurities.

Finally, Gump said, there is a need for greater transparency and more sharing of information within the industry. He sees long term opportunities to harmonize global approaches for dealing with this issue.

In July 2020, USP launched a broad-based outreach program to provide more direction to pharma manufacturers on how they might prevent supply chain vulnerabilities relating to nitrosamines. The organization released six new reference standards designed to support manufacturers and regulators in analyzing and monitoring potentially harmful NDMA and other nitrosamine impurities in the supply chain. USP will soon propose for public comment a new United States Pharmacopeia—National Formulary general chapter standard to provide guidance on assessing materials for nitrosamine presence, establishing control strategies for these impurities and ensuring the performance of analytical procedures to monitor nitrosamine levels in drug products.

Specifically, USP is proposing a new informational General Chapter <1469> Nitrosamine Impurities, which is aligned with current scientific and regulatory approaches for controlling nitrosamine impurities. This chapter provides a science-based approach for identifying and assessing the presence of these impurities in drug products in order to eliminate or reduce levels to help ensure quality as it relates to safety. General Chapter <1469> is scheduled for publication in September 2020 in USP’s online Pharmacopeia Forum, with the goal receiving stakeholder comments to proposed standards, says USP spokeswoman Theresa Laranang-Mutlu. The comment period will be open for 90 days, and stakeholders are strongly encouraged to provide input.

The chapter provides an introductory overview of concerns related to nitrosamine presence and identifies possible sources of nitrosamines in drug products along with observed or assessed risks associated with each source. It also includes a list of nitrosamines of recent concern in the pharmaceutical industry compiled from information shared by multiple global health authorities and input from members of USP’s Nitrosamines Joint Subcommittee.

The chapter would provide recommendations on nitrosamine risk assessment and the development of relevant control strategies to ensure that nitrosamine presence is avoided or below acceptable intake levels. A section on test method performance characteristics provides guidance on the verification process, procedures being implemented in the laboratory, and validation of alternative procedures.

The general chapter would also contain analytical procedures that have been validated or verified in USP’s laboratories and includes examples of quantitative analytical procedures. It also provides a compilation of procedures currently referenced on the websites of FDA and the European Directorate for the Quality of Medicines & HealthCare (EDQM), which users can verify as alternative procedures by meeting the requirements recommended in this chapter, says Laranang-Mutlu.

Focused on predictive tools and approaches, USP has gathered a group of volunteer experts to develop the nitrosamines general chapter, the USP Nitrosamines Joint Subcommittee, chaired by Mark Schweitzer, PhD, global head of analytical science and technology at Novartis, as well as representatives from FDA and the EDQM, which is responsible for drug quality standards in the European Pharmacopeia.

In August, USP will also be launching a new training webinar, “The ICH Q3 Impurity and the M7 Mutagenic Impurities Guidelines.” Designated as an ICH officially recognized training program, it will address the application of the ICH guidelines Q3-A, B, C, D and M7 for controlling organic, inorganic and solvent impurities. It will also aid professionals in developing strategies for dealing with actual and potential impurities that are most likely to arise during synthesis, purification, manufacturing, and storage of drug substances and drug products.

Practical steps: it’s not rocket science

In the meantime, there are some basic steps that companies can take every day to help prevent nitrosamine contamination, says Najafi, who has also been called as an expert witness in a number of cases and has seen scenarios of what can go wrong within a typical company when approaching this problem.

Ensuring that the right staff is involved in the problem is crucial, Najafi says. “The first step will be to employ experienced process development professionals and analytical chemists to oversee any outsourced manufacturing processes, specifically if changes or optimizations are being made at the partner site,” he says, “and it is important to evaluate the entire workflow with regard to use of amines, nitrosating agents, and recycled solvents, and to implement modern analytical testing for nitrosamines at strategic intervals.” He also suggests that the quality assurance department work in tandem with process chemistry and play a complementary role in preventing contamination. “This includes establishing Standard Operating Procedures for change control, frequent audits to ensure compliance, and launching investigations (e.g., during Phase I) when out-of-spec observations are made for impurities, and/or if unknown impurities are detected, however insignificant they seem,” he says.

“It is imperative that ‘for-profit’ corporations strive to increase revenue and implement changes to make manufacturing more efficient. However, the sponsor and the contract partner should collaborate on any such optimizations to ensure that efficiency does not come at the cost of patient safety. For example, recycling solvents is good for the environment and great for the company’s bottom line, however, there is a potential for carryover and new impurity introduction to an established manufacturing process from solvent reuse,” he says. Hence, the recycling process should be thoroughly validated, and stakeholders should incorporate extensive analytical tests for detection of unwanted contaminants that may result, he explains.

He notes problems that he has seen in the field. “Launching investigations is critical to ensuring process quality and compliance to regulatory guidance documents. One of most common errors I see is not conducting a thorough Phase I investigation when needed—companies either outright fail to launch one, do not properly document the process, or come to premature or faulty conclusions that are not backed by data or that lack scientific merit. I have seen all of these being committed by a company recently for a single manufacturing process,” he says.

Do not overlook cleaning and cleaning validation

It is also very important that contract manufacturers involved in multiple APIs and processes pay special attention to equipment cleaning to ensure that processes do not cross-contaminate each other. “Cleaning protocols should be validated, and suitable analytical tests performed to ensure that equipment is residue-free. In another recent example, I have seen a company implement an unvalidated cleaning strategy, and then apply an irrelevant analytical workflow to test for residues. Needless to say, that the company was cited by regulatory inspectors,” Najafi notes.

During the webcast, Najafi offered the following advice. First, to process development chemists: “If you intend to change or optimize manufacturing processes, look for potential development of NDMA. “If any of your processes contain an amine or nitrosating agent, be overly cautious and plan on testing for nitrosamines at every step,” he said.

Quality control departments, meanwhile, should be careful not to dismiss unknown peaks as “ghost” or “air” peaks, Najafi said, while quality assurance teams should be sure to conduct headspace gas chromatography and MS (HS-GCMS) on API routinely and identify all peaks. If the process is new, conduct HS-GCMS tests on the drug product as well, Najafi said.

“Aim to identify every unknown compound. Use HS-GCMS or liquid chromatography and mass spec (LCMS) if available and specifically look for NDMA and related nitrosamines,” he said. Attention to fundamentals is key, Najafi noted, cautioning manufacturing departments to remember that 96 ng of NDMA per pill can disqualify a batch “Proper cleaning of the equipment, and use of clean-in-place is important. One should be cognizant of potential cross-contamination problems,” he said.

Although Najafi joked, “It’s not rocket science,” on the webcast, it is clear that careful work and collaboration are key. New guidance and educational programs aim to address this need within the industry.

1. PharmTech, “Preventing Genotoxic Contamination: Lessons from the Nitrosamine Contamination Crisis,” Webcast, July 14, 2020.
2. A. Shanley, 

42 (10), pp. 60-64.
3. Emery Pharma Citizen’s Petition to FDA, 

, January 2, 2020.
4. A. Shanley, “Problems with Ranitidine May Transcend Manufacturing Issues,” 

, Oct. 2, 2019.
5. Valisure, Citizen’s Petition to FDA, 

, March 3, 2020.
6. ICH, ICH M7(R1), Assessment and Control of DNA-Reactive Impurities in Pharmaceuticals, 


Supplement: Outsourcing Resources 2020
Volume 44
August 2020
Pages: s18–s20

When referring to this article, please cite it as A. Shanley, " Collaborating to Prevent Genotoxic Contamination," 

Carcinogenic compounds have been found in a number of top-selling drugs, leading to recalls and bans. Preventing problems requires working closely with contract testing and development partners as well as API and other suppliers.

Collaborating to Prevent Genotoxic Contamination

	In 2019, cold-chain service providers continued investments in platforms and services designed for gene and cell therapy clinical studies. Marken acquired a logistics firm in Japan, and expanded logistics operations in Philadelphia with a new GMP-compliant plant and liquid nitrogen storage facilities, says president Ariette Van Strien. The company also launched a new European Communication Center of Excellence based in Edinburgh to provide communication, monitoring and management to thousands of investigator sites daily, she says, and opened a new cold-chain refrigeration-equipped facility in Dublin, Ireland. In India, Marken’s Direct-to-Patient (DTP) distribution services are now actively delivering to and from patients in 94 cities, something that had not been permitted in India prior to the outbreak of COVID-19. However, given the critical need to keep patients in their clinical trials, the government has now permitted the service to proceed. Marken also introduced Smart Box packaging for temperature-critical shipments, says Van Strien. 

	World Courier introduced a complete cryogenic supply chain, which offers fully automated technology and temperature-controlled transport from the manufacturer’s location to a storage facility and then to each point of care in dry shippers. 

	Through a recent integration with the third-party logistics company ICS, World Courier can charge dry shipments across its network of more than 140 offices and depots, says Alex Guite, vice-president of services and alliances for World Courier. 

	Before the pandemic, Catalent had introduced solutions designed to enable greater flexibility and responsiveness to change, including demand-led and direct-to-patient supply programs, says Amy Lombardi, group product manager for Catalent’s clinical supply services. The company has been expanding its cold chain capacity for some time, especially in the Asia Pacific region, she says. “In the face of the COVID-19 pandemic, these services have proved to be important tools that allow customers to adapt their ongoing studies and keep medication flowing to patients, even when they cannot or will not leave their homes to visit a clinical site,” says Lombardi. 

	Almac made some important changes in 2019, in response to a spike in demand for end-to-end temperature-controlled programs, especially in relation to closing temperature data gaps that often exist at clinical sites, says Sharon Courtney, logistics services manager for Almac clinical services 

	“Although effectively managing and monitoring temperature during production, storage, and global distribution of clinical supplies can be achieved through appropriate facilities, monitors and shippers, it’s surprisingly common for temperature data visibility challenges when products arrive at sites. This can be largely due to a reliance on manual data capture that makes accessing records difficult, creating risk and leaving sponsors struggling to identify and adjudicate site-based excursions, assess any trends, maintain compliance and safeguard patients,” says Courtney. 

	In response, Almac recently launched a new module to its TempEZ platform (Site Storage Temperature Compliance) that plugs the data gap by digitalizing temperature management at site level to increase monitor data upload rates and promote compliance, without adding to the administration burden. A new European Campus in Ireland is designed to help sponsors grapple with the potential challenges of operating clinical trials in the European Union post Brexit, Courtney says. 

	When referring to this article, please cite it as A. Shanley, "2019 Investments entered Around Cell and Gene Therapies," 

Previous investments set a foundation for later efficiency improvements. 

2019 Investments Centered around Cell and Gene Therapies

Since traces of the compounds were first found in batches of APIs used to make valsartan and other sartan ARBs in 2018, drug manufacturers have been paying much closer attention to nitrosamines -in particular NDMA (N-Nitrosodimethylamine)-and how they enter the pharmaceutical supply chain. “Nitrosamines are extremely carcinogenic, as academic researchers have known for decades. The fact that NDMA is used to this day as a control to induce cancer in rats just underscores its immense carcinogenic potential,” says David Light, CEO of Valisure, an analytical pharmacy that filed the first citizen’s petition on nitrosamine contamination in ranitidine with FDA.  

	Some cases of contamination have been traced to solvents and to inadequate manufacturing and quality testing practices. In the case of the first contaminated API, for valsartan, one supplier had tweaked its synthesis process to improve yield and efficiency. Not only the manufacturer but the regulators who okayed that change failed to realize that it would result in contamination (1). 

	But ARBs were only the first sign of trouble, and, for one drug (so far), the problems go much deeper than manufacturing and quality control.  In 2019, extensive testing found that ranitidine, one of the 

most widely prescribed treatments for ulcers and acid indigestion 

 (whether as the trade named Zantac or as one of many generic and over-the-counter brands), was also contaminated with NDMA. Valisure and Emery Pharma filed citizen’s petitions with FDA in 2019 and 2020. detailing the public health dangers posed by this contamination (2 - 4).  Although manufacturing and storage practices have led to contamination in some cases, the ranitidine molecule itself was found to be inherently unstable, breaking down and generating NDMA in the presence of high temperatures and humidity levels. After disputes about the best methods to use for analysis and testing (4), product recalls ensued, and on April 1, 2020 the US FDA followed other global regulatory agencies in banning the sale of ranitidine and products that contain the compound (5). This product had been on the market for almost 40 years and was widely prescribed to pregnant women and young children. 

	In December 2019, news came from regulators in Singapore, and later, from Canadian and Swiss health authorities that NDMA and nitrosamines had also been found in metformin, the fourth most widely prescribed drug in the US, and one of the most widely prescribed diabetes treatments in the world. In March 2020, after testing 38 lots of metformin from 22 suppliers, Valisure filed a citizen’s petition with FDA (6).

	Far from being isolated instances, nitrosamine contamination is a systemic issue that manufacturers of all types need to take seriously. Although more is being learned about how it occurs, there are still gaps in that knowledge, says Ron Najafi, CEO of Emery Pharma, which sent a citizen’s petition to FDA in January 2020 after studying ranitidine stability to identify the source of NDMA contamination. Emery is testing a number of other pharmaceuticals in an attempt to clarify the root causes of such contamination. The company has already developed and validated new analytical methods for such tests. 

	The United States Pharmacopeial Convention (USP) will be releasing new guidance later in 2020 to help manufacturers better face, address, and prevent nitrosamine contamination problems for pharmaceuticals and APIs. Included will be an informational chapter, as well as a series of six reference standards that can be used as controls and in method validation, says Jaap Venema, chief scientific officer with USP. 

	Venema believes that decisive action is required now, since the contaminant has been found in three very different types of products from very different sources.  Longer term, he believes that more work is needed in assessing the risk, not only of nitrosamines, but of other genotoxins. “We need to do more as a healthcare system, as a supply chain, to control, test and predict potential impurities,” he says.

	Considering the first recent contamination case involving the valsartan API, initially there was a lot of finger pointing and blaming of the manufacturer, a Japanese company doing business in China, in media reports. In fact, Venema emphasizes that, although manufacturing facilities and processes were inspected, no one expected to find NDMA in that setting.  “If there is one lesson to be learned by all, it is to expect the unexpected…It all comes down to the need for more control, more testing, and more predictive work,” says Venema. 

	After other global regulatory agencies recalled metformin, FDA tested samples of the drug, examining material from 16 batches made by seven manufacturers.  The agency released data in February 2020 that found contamination levels below the limit, Light recalls.

	Valisure decided to investigate metformin after a person sent Valisure a sample of the compound and said that she was concerned about its purity, Light says. Valisure’s lab found a very high level of NDMA in that sample, he says. Valisure then bought more metformin from distributors, analyzing 38 batches of the material from 22 companies. During testing, 16 of the batches failed. Half of the manufacturers had at least one batch that failed, while many had batches that contained more than ten times the legal limit of NDMA, Light says (7).

	These results highlight some of the procedures that limit FDA, which, when it tests for contamination, must ask the industry for voluntary samples. “Manufacturers can 

 choose what they send. We buy direct from distributors, rather than manufacturers, so we get material from the end of the supply chain,” he says. Light filed the Citizen’s Petition on March 2.  

	There have been ongoing questions about the best analytical methods for detecting nitrosamines in pharmaceuticals. “I don’t think there will ever be a consensus among stakeholders as to a single set of ideal conditions or methods. Instead, I believe drugs should be tested by multiple complementary methods in an unbiased fashion, and the conditions/tests set on a case-by-case basis,” says Light.

	As he notes, disagreements that ensued after NDMA detection in ranitidine using headspace GC-MS (‘HS-GC-MS’) have largely been resolved and it is now well-established that HS-GC-MS should not be the method for analysis of ranitidine. “As careful scientists looking at the problem as a whole, we developed our own low-temperature GCMS version to analyze ranitidine. We also thought the 15 min exposure to 130 º C had a dramatic effect on ranitidine.  As chemists, we saw that it was a very efficient reaction, to have a nearly 10 % molar conversion in 15 minutes of 130 º C exposure, most other drug molecules do fine under those conditions. Which is why that method had been the standard.

	From a scientific perspective it doesn’t really matter what method you use as long as you address primary issues. We addressed molecule’s instability by using lower temperatures with GC-MS, FDA did the same for LC-MS.“There is a need to think a little bit outside of the box. You cannot wait for there to be a very specific prescribed system for the exact drug you’re looking at. If we had done that, rantidine would still be on the market around the world,” he says.

	Emery has collaborated with Valisure on much of its work, and corroborated its test results.  The company explored use of alternative methods, including liquid chromatography (‘LC’) – tandem mass spectrometry (‘MS/MS’), which does not expose the samples to high temperatures.  Using this method, together with stability studies, Najafi and his team found that ranitidine is an unstable compound, a time-and temperature-sensitive pharmaceutical product, that breaks down to form NDMA.

	“After nearly four decades in the market, we were the first to prove that ranitidine develops the carcinogen NDMA when exposed to heat, a common occurrence during transportation and storage,” says Najafi.  Therein lies the importance of Emery’s findings, he says, since temperature excursions are quite common during commercial pharmaceutical shipment.

	Emery had a strong set of preliminary data (unpublished at this point) to support these claims, Nafaji says, when, on December 4, 2019, FDA offered a directive for manufacturers regarding ranitidine product release. The Agency said that distributors and manufacturers could release ranitidine drug products back into the market, so long as production batches showed NDMA levels lower than 96 ng/pill (the daily acceptable limit of NDMA). “We were extremely concerned, because no formal root-cause analyses had been performed to determine why NDMA was observed in several lots of Zantac and other ranitidine drug products,” says Najafi. 

	“Even if manufacturing lots were clean, NDMA could still be produced in ranitidine drugs products during downstream shipment and storage, or if stored by patients under less-than-ideal conditions, e.g. in a car dashboard,” says Najafi. “We felt an obligation to inform the agency of our findings, given the major implications they had for public health,” he says, and filed a citizen’s petition with FDA in January 2020.

	ICH M-7 is a comprehensive guidance for genotoxic compounds that has guided FDA’s approach to nitrosamine contaminants. Kristi Muldoon Jacobs, USP’s regulatory science director, likes the fact that it allows pharma to take advantage of advances in toxicology and collective knowledge, to help identify genotoxic impurities. 

	However, there are some places where it could be improved or added to, she says. For example, when discussing known and suspected mutagens, some chemicals, for which there isn’t enough information (and which include nitrosamines) are labeled “cohorts of concern.” The guidance doesn’t give much information on how those compounds should be addressed, so the pharma community should add to the guidance as more is learned from specific cases, she says. For example, there is a list of chemicals for which acceptable daily intake (or ADI) levels have been published, and now that more information has been published on nitrosamines, it can be used to update the guidance, she says. 

	Current FDA and USP guidance is reasonable, albeit a bit dated at this point, says Najafi.“Impurities are often better dealt with on a case-by-case basis, which is difficult to do with a broad guidance document,” he says. One problem that he sees is the bureaucratic red tape that surrounds updating these documents. “We need quicker, more facile updates to guidance documents and monographs, and analytical methods from 36 years ago should be updated with newer instrumentations and methodologies,” he says.  For examples, manufacturers could be required to perform unbiased assessment of impurities in every production lot, using multiple analytical methodologies (e.g., GC-MS, LC-MS, UV-Vis, NMR, or HPLC) instead of being allowed to pick and choose their favorite method., says Najafi, then they could pursue all new or unidentified peaks, identify the impurities/analytes responsible for these peak(s), and ensure these are within established safety profiles before releasing a product lot.  “On the other hand,” Muldoon Jacobs says, “standards ensure that the methods are fit for purpose and that everyone is using the same methods.  USP is currently working to update and providing additional chapters to provide risk-based approaches and suitable verifiable testing methods.”

	Greater familiarity of and compliance with FDA, EMA, ICH, guidance is needed, Najafi says. “Regulators around the globe need to come down on non-compliance with an iron fist,” he adds. He points to the 2018 valsartan API contamination, when problems came up when manufacturers deviated from monographs and established procedures to improve process yield.  Generic pharmaceutical manufacturers, especially those offshore, should be required to stick with the monograph. “If a recipe calls for having a fly in the room, you need to have one there,” says Najafi.

	During the new drug application (NDA) phase, CMC (chemistry, manufacturing and control) review is very tough, but generic pharmaceuticals don’t undergo that same level of scrutiny. If a generic drug manufacturer wants to make process changes, Najafi says, they should submit an application for re-approval to FDA, even if they’re only making a change as basic as switching solvents. “A contamination level of 96 ng per pill may not be a lot, in fact, it is less than 0.00001%, but NDMA is so significant a toxic compound that one should worry and have control over it,” says Najafi.

	Manufacturers also need to do better on testing, particularly with residual solvent analysis, says Najafi, and manufacturers should be able to analyze, record, and report levels of each solvent. In reality, some manufacturers completely ignore these impurities, he says. “There is no reason to ignore them. We have amazing analytical instruments such as mass spectrometers to perform the analyses. If manufacturers argue that they can’t afford a GC-MS system, they shouldn’t be in business,” he says. 

	Scientists working in other industries have been studying nitrosamine contamination since the 1970’s, says Light, but pharma and its regulators have often seemed disconnected from that research.  In addition, he says, there is a very large body of research on the potential risks of nitrosatable drugs (e.g., those containing secondary and tertiary amines, in which the building blocks of nitrosamine carcinogens are on the drugs themselves). A number of studies have examined nitrosatable drugs during pregnancy, and correlated use of the drugs with teratogenic effects, stillbirths, and tumors, says Light. 

	“The pharmaceutical world has not kept up with other industries on the topic of nitrosamines, although it is getting better,” says Najafi. “There is a wealth of literature on the subject from scientists in the food processing and waste-water industries that we in the pharmaceutical world must continue to tap into,” he says.“Widespread cross-talk among scientist is the only way to bridge the existing knowledge gap, and government agencies and regulators should facilitate it,” says Najafi. Emery, for example, has started collaborating on studies of nitrosamine and other genotoxic impurities with Prof. Bill Mitch from Stanford University, a specialist in the field of environmental biomonitoring, public health, and emerging carcinogens.

	In the meantime, the toxicological methods used to study nitrosamine impurities in pharma  continue to evolve. “I believe this area is still a black box - long-term exposure risk is and will remain challenging to evaluate,” says Najafi. “While working on ranitidine, we realized that there have been no clean epidemiological studies on this drug, even though ranitidine has been on the market for decades and has been taken by millions around the world! The same may be true for many more drugs,” he says. Valisure is working with 

on a study of ranitidine’s health effects, which underwent peer review and was scheduled to be published in January 2020 in the 

Journal of the American Medical Association

, but which is being held for further review. Light expects the article to be published at some point this year.

	The field of toxicology is advancing rapidly, adopting more predictive and 

 approaches, says Muldoon Jacobs. In the future, she expects predictive tools to be routinely pulled into manufacturing, to predict whether impurities may be present and to help determine their potential toxicity. 

	Some computer-based predictive models, she says, can be used to predict whether a compound’s or contaminant’s given chemical structure might be mutagenic and whether or not it would test positive in the Ames assay (8), which has been used for over 25 years to identify mutagenic compounds. “These models are built on hundreds or thousands of individual data pieces based on actual studies, and some scientific papers have shown that these models are more accurate than the Ames test,” she says.  Pharmaceutical industry regulatory agencies are evaluating and applying these methods, for instance, at FDA’s Division of Applied Regulatory Science, she says.

	Although mass spec and hyphenated gas and liquid chromatography methods are being used to detect nitrosamines in the pharmaceutical supply chain, new analytical methods may be applied in the future.  The UK-based company, Ellutia, for instance, offers TEA detector technology which uses chemiluminescence at the GC or LC interface, to measure levels of N-nitrosamines. A Total analyzer is available that gives one cumulative measurement of all nitrosamine contaminants in four minutes. The detector is widely used in Europe in the rubber toy, brewing, water treatment and food industries, but pharmaceutical companies have just begun to explore its potential. 

	The industry is taking the issue of nitrosamine contamination and its potential human health impacts more seriously. “As with the current COVID-19 pandemic, it’s not a matter of if, but when, situations like this will come up. Nitrosamines are very much on our minds right now, but history has taught us there will be other cases involving other genotoxic contaminants in the future. The questions for the industry now are: How do we prevent more of these situations from occurring (knowing there won’t be a preventive method that is 100% fail-safe)? How do we prepare for them, and how do we respond faster and more responsibly when they recur?  The real lesson for our industry is to get better at predicting and preventing these problems,” says Venema.

42 (10), pp. 60-64 (2018).
	2. Valisure, 

, January 2, 2020.
	4. A. Shanley, “Problems With Ranitidine May Transcend Manufacturing Issues,” 

, October 2, 2019.
	5. FDA News Release, “FDA Requests Removal of All Ranitidine Products From the Market,” 

Statement from Janet Woodcock on Nitrosamine Impurities Found in Diabetes Drugs Outside the US, fda.gov

, March 2, 2020, valisure.com
	8. R. Luhana, “Online Pharmacy Valisure Discovers Carcinogen in Some Batches of Metformin,” 

Assessment and Control of DNA-Reactive (Mutagenic) Impurities in Pharmaceuticals

	Pharmaceutical Technology
	Vol. 44, No. 5
	May 2020
	Pages: 46-49

	When referring to this article, please cite it as A. Shanley, "Contamination Drives a More Concerted Approach to Genotoxins," 

Discovery of carcinogenic nitrosamines in three of the world’s most widely prescribed drugs is driving efforts to better detect, control, and prevent their generation in APIs and finished drug products.


Contamination Drives a More Concerted Approach to Genotoxins

Discovery of carcinogenic nitrosamines in three of the world’s most widely prescribed drugs is driving efforts to better detect, control and prevent their generation in APIs and finished drug products.
