Cell and gene therapies are rapidly emerging as promising alternatives to treat patients with severe forms of cancer, particularly hematological malignancies, due to their targeted efficacy and high potency. These types of therapies are often referred to as advanced therapeutic medicinal products (ATMPs) and are gaining traction in the public arena as a result of FDA’s approval of chimeric antigen receptor (CAR) T-cell therapies. Six different CAR T-cell therapies are now approved by FDA for the treatment of certain types of lymphomas, some forms of leukemia (1), and most recently, multiple myeloma (2).

CAR T-cell therapies involve genetically engineering a patient’s own or a donors’ T-cells to target and destroy cancer cells. They are highly complex and potentially risky treatments that are showing clinical and commercial success where other available treatments have failed.

Due to the specialized nature of CAR T-cell therapy and the vulnerability of patients with compromised immune systems, sterility is essential for these treatments. As CAR T-cell products are made up of living cells, it is not possible to perform sterilization (e.g., sterile filtration) of the final product. Therefore, safe and effective CAR T-cell therapies rely on rigorous sterility testing to ensure patient safety and product quality.

Sterility testing of autologous cell therapies

In autologous cell therapies, such as CAR T-cells, sterility testing is an essential and critical step in the batch release process under the Code of Federal Regulations, Title 21, parts 210 and 211 for current good manufacturing practices (CGMP) for finished pharmaceuticals and parts 600 to 680 for additional biological product standards (3). Assurance of final product sterility also relies on procedural measures that effectively prevent contamination of biological materials, such as cleanroom technology and other GMP standards.

The compendial sterility test is recognized by the industry for its adherence to regulatory standards and for its reliability. Compendial testing involves direct inoculation of the CAR T-cell sample into two different media (4), for growth of aerobic and anaerobic microorganisms, followed by incubation at different temperatures. During incubation, visual examination of media is performed by checking turbidity. However, the final examination of the medium for evidence of microbial growth occurs after a 14-day incubation period. This extended lead-time can be challenging for patients in urgent need of treatment once the drug product is manufactured, particularly if the final formulation cannot be cryopreserved and has a short shelf-life. The limited amount of a sample, combined with the time sensitivity and critical need of severely sick patients, means a faster sterility method is required.

The development of more rapid sterility testing methods offers significant advantages. It can be beneficial for frozen CAR T-cell products, expediting the time from product manufacture to patient administration, which is crucial in the treatment of severely sick patients in the late stages of disease. To ensure the deployment of safe and effective therapies to patients, the industry needs to evaluate and promote effective rapid sterility methods.

When referring to this article, please cite it as Doláková, M.; Horková, V.; Sochorová, K.; Rožková, D. Rapid Sterility Methods for CAR T-cell Therapies. 

Articles

A combination of rapid sterility methods with the industry standard compendial method should ensure maximum safety.

Rapid Sterility Methods for CAR T-cell Therapies

Mathematic calculations are a critical activity in pharmaceutical compounding. All compounded preparations will require calculations at some time during their lifecycle. Types of calculations are many and varied—basic math, drug concentration, dose adjustments, batch size changes, stability testing, and many others—all of which require human performance.

This discussion describes the following series of activities to execute calculations for preparation of compounded dosage forms:

Stage 1 equation design. Work prior to actual calculations; integrating information, resolving inconsistencies, equation set-up, and initiating documentation.

Stage 2 numeric operations. Equation execution, result evaluation, and completing documentation.

Stage 3 independent review. Verification of information, calculations, and documentation

Management responsibilities. Key activities, personnel, priorities, and workplace culture.

Activities described address performance in the workplace environment. These may be performed by an individual person in a 503A pharmacy or by multiple people in 503B outsourcing facilities, pharma industry, and other settings.

Content in this discussion reflects academic teaching, personal experiences in multiple organizations, and published accounts of adverse events. Textbooks (1–9) and academic education provide theory. Application in the workplace is much different—actual drugs, sick patients, medication orders in multiple formats, time pressures, information discrepancies, and workplace interruptions and distractions—all of which have potential for errors. Calculations errors may have significant ramifications: patients receive toxic overdoses or ineffective underdoses; healthcare practitioners suffer personal remorse (10); management is embarrassed (11); and organizations face legal consequences (12).

Additional discussion on this topic including case studies is available under the same title at PharmTech.com (13). The calculations process in this discussion is focused on specific calculations activities separate from a comprehensive dosage form compounding process.

Stage 1, equation design, comprises activities in advance of actual calculations (i.e., thinking, planning, verifying information, estimating, setting up equations, and initiating documentation). Stage 1 emphasizes proactive thinking about the calculations to be performed.

Calculations objective. Calculations for preparation of a compounded dosage form must be clearly identified. Three questions are fundamental:

What is to be calculated? What is the desired final outcome of the calculations? Where are we going?

What information is known? What is given, what is relevant, what is not relevant? What is the medication order telling us? Excess information sometimes confuses identifying the calculation objective.

What is unknown? What don’t we know, what is missing, what is questionable and needs confirmation, and how do we get this information?

This is a tried-and-true process in problem solving, troubleshooting, investigations, and related activities. It forces thorough and logical thinking. A plan to accomplish the calculations objective is needed. One cannot begin to set up equations without initiating logical and common-sense thought. Especially relevant in these activities are the associated units of measure for all values.

Compile information. Concurrent with identification of the calculations objective is the collection of relevant information. Missing information must be obtained; questionable information must be investigated. Pertinent information must be written; do not make assumptions or depend on memory. Correctly executed calculations using erroneous information may be fatal (14). Verbal information must be repeated and confirmed in writing. Misunderstanding due to language dialects may contribute to errors.

Paul L. Pluta, RPh, PhD, is a pharmaceutical scientist. Alan M. Mancini, RPh, is a pharmaceutical scientist. Nishant B. Thakar, RPh, PharmD is assistant professor Clinical Sciences, Roosevelt University College of Science, Health, and Pharmacy. Varanya Chaiyaperm, RPh, PharmD is clinical assistant professor, University of Illinois College of Pharmacy.

When referring to this article, please cite it as Pluta, P.L.; Mancini, A.M.; Takar, N.B.; Chaiyaperm, V. Pharmaceutical Compounding Calculations in a Workplace Environment. 

Pharmaceutical Technology Trends in Manufacturing

Manufacturing

An ordered process describing calculations activities for compounded dosage forms is described.

Pharmaceutical Compounding Calculations in a Workplace Environment

Multimodal manufacturing is a strategic approach that harnesses the power of diverse manufacturing methods to optimize and streamline the production process, specifically within the realm of human therapeutics in the advanced therapy medicinal products (ATMP) space. This concept involves integrating different manufacturing techniques/platforms to improve efficiency, reduce costs, and enhance the overall quality of the manufacturing process and the eventual therapeutic product slated for licensure. 

In a White House report, Bold Goals for US Biotechnology and Biomanufacturing, a prominent objective is to test and de-risk new biomanufacturing practices for next-generation biotechnology products in commercial manufacturing facilities (1). The growing focus in cell-based therapies, a key component of the next generation of human therapeutics, has led many organizations to focus their development efforts into a multitude of platforms.

Multimodal is not new. For the ATMP space, it was identified back in 2017 as a viable manufacturing option for clinical needs. In the context of the biotechnology ATMP space, multimodal manufacturing techniques are being applied to optimize the production of complex biological compounds and therapies that are studied in multi-omic research, such as gene editing (1).

In reality, many contract development and manufacturing organizations have been operating in a quasi-multimodal manner for decades. Multimodal can include vaccines, monoclonal antibodies (mAbs), viral vectors, plasmids, allogeneic cell therapies, and autologous cell therapies. The challenge is crafting a manufacturing plan that supports one’s business without attempting to become a one-size-fits-all solution for every need.

The forecasted growth of the cell and gene therapy (CGT) market is significant. The ATMP space, where CGT continues to serve as the primary catalyst for growth, is predicted to grow at a compound annual growth rate of 39.4% for the period from 2024 until 2030 (2).

In this growth projection, cell therapies account for 15% of the growth and gene therapies account for another 30% of growth, with many of these products advancing out of early-stage clinical manufacturing toward commercialization (2).

Just as the 1990’s large-molecule market saw a “fork-in-the-road” decision regarding single vs. multi-product manufacturing and the technologies to implement (3), the ATMP space is grappling with critical questions around asset size, cost, technology implementation, flexibility, scale, and many others. Transitioning to a multimodal manufacturing strategy could very well emerge as the most prudent business and operational solution.

Another key driver fueling the multimodal manufacturing discourse is the rapid pace of advancement in research, developing different modes of therapy to address multiple human therapeutic targets. Combination therapies using multiple therapeutic modalities have elevated anti-cancer activity while lowering doses of agents, thus reducing side effects (4). As previously referenced, gene editing tools such as clustered regularly interspaced palindromic repeats (CRISPR)-Cas, transcription activator-like effector nucleases (TALENs), and base editing have brought about multiple therapeutic solutions that are being provided for chronic diseases and oncolytic cancer treatments, as well as advanced chimeric antigen receptor T cell (CAR-T) and T-cell receptor (TCR) therapies.

Pharmaceutical Technology’s Trends in Manufacturing May 2024 eBook

1. The White House. Bold Goals for US Biotechnology and Biomanufacturing: Harnessing Research and Development to Further Societal Goals. White House, Executive Order 14081, March 2023.
2. Vision Research Reports. Cell and Gene Therapy Market (By Therapy Type: Gene Therapy, Cell Therapy; By Application: Oncology, Genetic Disorders, Dermatology Disorders, Musculoskeletal Disorders, Others; End-Users: Hospitals, Cancer Care Centers, Wound Care Centers, Others)—Global Industry Analysis, Size, Share, Growth, Trends, Regional Outlook, and Forecast 2022-2030. Market Research Report, July 2022.
3. Witcher, M.F.; Carbonell, R.;Odum, J.; et al. Facility of the Future: Next Generation Biomanufacturing Forum Parts I, II, III. 

Jeff Odum, CPIP, is practice lead, ATMPs & Biologics, at Genesis AEC.

When referring to this article, please cite it as Odum, J. Exploring Multimodal Biomanufacturing and Its Drivers. 

Exploring Multimodal Biomanufacturing and Its Drivers

The COVID-19 pandemic has given the bio/pharma industry lessons on being prepared to manufacture parenteral formulations for human use in emergency situations. Vaccines, remdesivir injection and amphotericin injection, are such recent examples. The challenge then becomes having enough flexibility in the manufacturing lines to facilitate manufacture of large volumes of unit doses of injectables as may be required with shorter changeover times, while assuring compliance and patient safety from the product contact parts used in the manufacturing line. Single-use manufacturing solutions provide the flexibility in terms of batch sizes, fill volumes, and ease of manufacturing, especially in terms of manufacturing different products in the same line with minimum line changeover requirements. This article reviews and elaborates the initial risk assessment process for extractable and leachables arising from single-use product contact manufacturing parts as per 

) chapters <665> and <1665>, which go into effect May 1, 2026 (1,2). The determined risk can then be used to build the level of extractable profiling and process equipment-related leachables (PERLs) followed by assessing the patient safety for regulatory submissions. 

Manufacturing systems used for biopharmaceutical drug substances or drug products can be partially or completely constructed from plastic materials. During the manufacturing process, the drug product and its components could directly come in contact with the plastics and potentially interact with one or more of the manufacturing system’s product contact components. The substances that leach from a product contact manufacturing component could become incorporated into the process stream. If these leached substances persist in the process stream and continue through the process operations, they could accumulate in the drug substance or drug product as PERLs (1,2). When present in the drug product, these PERLs can potentially alter the critical quality attributes of the drug product, such as its stability, efficacy, and safety. 

Plastic Components and Systems Used to Manufacture Pharmaceutical Drug Products and Biopharmaceutical Drug Substances and Products

, addresses these interactions by providing a risk-based means for chemically characterizing and qualifying plastic components used to manufacture biopharmaceutical drug substances and pharmaceutical and biopharmaceutical drug products. 

Characterization and Qualification of Plastic Components and Systems Used to Manufacture Pharmaceutical Drug Products and Biopharmaceutical Drug Substances and Products

, communicates the key concepts behind and provides additional information and guidance regarding the applicability and the application of 

 <665>. The authors discuss a practical approach by which 

 <1665> can be applied in a step-by-step manner for the risk assessment, using a case study involving the manufacturing process for remdesivir injection and single-use technologies (Merck).

The testing of a product contact component is driven by the risk that the component may be unsuited for its intended use based on chemical or biological effects attributed to PERLs (1). As such risks increase, the degree of required testing also increases. The risk assessment process is accomplished by a risk evaluation matrix that establishes the appropriate contributors to the dimension of risk, provides a quantifying means for the risk in each dimension, quantifies the total risk for each dimension, and links the quantified risk to an appropriate characterization strategy.

 <665>, Plastic Components and Systems Used to Manufacture Pharmaceutical Drug Products and Biopharmaceutical Drug Substances and Products. 

 <1665>, Characterization and Qualification of Plastic Components and Systems Used to Manufacture Pharmaceutical Drug Products and Biopharmaceutical Drug Substances and Products. 

Ramesh Raju Mavuleti*, rrajum@gmail.com, is head of operations, Validation Services India. Subhasis Banerjee, PhD, is principal technical application expert, Bioprocessing APAC. Tathagata Ray is consultant, Global Strategy Deployment; all at Merck Life Sciences Pvt, Ltd. K. Vasantakumar Pai, PhD, is professor and chairman, PG Department of studies & research in Industrial Chemistry, Kuvempu University. K. Sreedhara Ranganath Pai, PhD, is professor, Department of Pharmacology, Manipal Academy of Higher Education. Somasundaram Gopalakrishnan is senior consultant, Global Biopharm Center of Excellence, Merck Pte. Ltd.

*To whom all correspondence should be addressed.

When referring to this article, please cite it as Mavuleti, R. R.; Pai, K. V.; Pai, K. S. R.; et al. Application of the Risk Evaluation Matrix as per 

<665> and <1665> for Evaluation of Leachables Risk from Single-Use Components Used in Biomanufacturing. 

Quality

Determining E&L risk from single-use components can be used to build the level of extractable profiling and PERLs.

Application of the Risk Evaluation Matrix as per USP <665> and <1665> for Evaluation of Leachables Risk from Single-Use Components Used in Biomanufacturing

Trends in Manufacturing, May 2024 eBook

Rapid Sterility Methods for CAR T-cell Therapies

Pharmaceutical Compounding Calculations in a Workplace Environment

Exploring Multimodal Biomanufacturing and Its Drivers

Application of the Risk Evaluation Matrix as per USP <665> and <1665> for Evaluation of Leachables Risk from Single-Use Components Used in Biomanufacturing