Regulatory Acceptance Research Articles

Making Markets

We examine the impact of design on the evolution of a product market from illicit to mainstream. We argue the importance of congruence with normative and cultural-cognitive structures in fostering legitimacy. To understand the role of product design in this process, we conducted an ethnographic study of the newly-legal recreational cannabis market in the US, a market that has attained regulatory acceptance in some states but lacks normative and cultural-cognitive legitimacy. By analysing product design and interviewing managers, we find that design plays a pivotal role in legitimation. Producers transform a market by manipulating two distinct aspects of materiality: material in relation – how products relate to accepted products, and material in use – how design guides and enables consumption. We offer a framework for managing products in new markets, arguing that design can enable legitimacy by drawing on symbolic relationships to other products, considering affordances, and enhancing strategic socio-cultural innovation.

Production of selectable marker gene-free Cavendish banana (Musa spp.) using a steroid-inducible recombinase platform

Genetic improvement of commercially accepted banana cultivars is strongly reliant on the ability to introduce genes that encode important agro-traits such as disease resistance. In most cases this can only be achieved using a transgenic approach. Public and regulatory acceptance of these events would greatly increase with “clean” single copy integration events free of the selectable marker gene and extraneous vector backbone. This would also allow for the successive addition of new genes and traits as they become available. In this study, we used the pMarker Free 1 (pMF1) vector containing the green fluorescent protein (gfp) reporter gene to assess the effectiveness of steroid-inducible recombination and positive/negative dual selection to regenerate transgenic Cavendish banana plants that were potentially free of the selectable marker gene. By examining the interaction of two different Agrobacterium strains with two different cultivars of Cavendish banana, namely Williams and Grand Naine, we describe a transformation and regeneration strategy that successfully produced marker-free, single transgene copy, gfp-expressing events. The system will provide a useful means of serially improving banana into the future.

Development of In Vitro\u2013In Vivo Correlation for Upadacitinib Extended-Release Tablet Formulation

Upadacitinib is a selective Janus Kinase 1 inhibitor which is being developed for the treatment of several inflammatory diseases including rheumatoid arthritis. Upadacitinib was evaluated in Phase 3 studies as an oral extended-release (ER) formulation administered once daily. The purpose of this study was to develop a level A in vitro–in vivo correlation (IVIVC) for upadacitinib ER formulation. The pharmacokinetics of four upadacitinib extended-release formulations with different in vitro release characteristics and an immediate-release capsule formulation of upadacitinib were evaluated in 20 healthy subjects in a single-dose, randomized, crossover study. In vivo pharmacokinetic data and in vitro dissolution data (USP Dissolution Apparatus 1; pH 6.8; 100 rpm) were used to establish a level A IVIVC. Three formulations were used to establish the IVIVC, and the fourth formulation was used for external validation. A non-linear IVIVC best described the relationship between upadacitinib in vitro dissolution and in vivo absorption profiles. The absolute percent prediction errors (%PE) for upadacitinib Cmax and AUC were less than 10% for all three formulations used to establish the IVIVC, as well as for the %PE for the external validation formulation and the overall mean internal validation. Model was cross-validated using the leave-one-out approach; all evaluated cross-validation runs met the regulatory acceptance criteria. A level A IVIVC was successfully developed and validated for upadacitinib ER formulation, which meets the FDA and EMA regulatory validation criteria and can be used as surrogate for in vivo bioequivalence.

An Institutional Approach to Gender Diversity and Firm Performance

This study examines data from 35 countries and 24 industries to understand the relationship between gender diversity and firm performance. Previous studies report conflicting evidence: some find that gender-diverse firms experience more positive performance, and others find the opposite. However, most research to date has focused on a single country or industry and has not accounted for possible variation across social contexts. This paper advances an institutional framework and predicts that gender diversity’s relationship with performance depends on both its normative and regulatory acceptance in the broader institutional environment. Using a unique longitudinal sample of 1,069 leading public firms around the world, I find that the relationship between gender diversity and firm performance varies significantly across countries and industries owing to differences in institutional context. The more that gender diversity has been normatively accepted in a country or industry, the more that gender-diverse firms experience positive market valuation and increased revenue. These findings underscore the importance of the broader social context when considering the relationship between gender diversity and firm performance.

Metadata Concepts for Advancing the Use of Digital Health Technologies in Clinical Research

Digital health technologies (smartphones, smartwatches, and other body-worn sensors) can act as novel tools to aid in the diagnosis and remote objective monitoring of an individual’s disease symptoms, both in clinical care and in research. Nonetheless, such digital health technologies have yet to widely demonstrate value in clinical research due to insufficient data interpretability and lack of regulatory acceptance. Metadata, i.e., data that accompany and describe the primary data, can be utilized to better understand the context of the sensor data and can assist in data management, data sharing, and subsequent data analysis. The need for data and metadata standards for digital health technologies has been raised in academic and industry research communities and has also been noted by regulatory authorities. Therefore, to address this unmet need, we here propose a metadata set that reflects regulatory guidelines and that can serve as a conceptual map to (1) inform researchers on the metadata they should collect in digital health studies, aiming to increase the interpretability and exchangeability of their data, and (2) direct standard development organizations on how to extend their existing standards to incorporate digital health technologies. The proposed metadata set is informed by existing standards pertaining to clinical trials and medical devices, in addition to existing schemas that have supported digital health technology studies. We illustrate this specifically in the context of Parkinson’s disease, as a model for a wide range of other chronic conditions for which remote monitoring would be useful in both care and science. We invite the scientific and clinical research communities to apply the proposed metadata set to ongoing and planned research. Where the proposed metadata fall short, we ask users to contribute to its ongoing revision so that an adequate degree of consensus can be maintained in a rapidly evolving technology landscape.

Organotypic and Microphysiological Models of Liver, Gut, and Kidney for Studies of Drug Metabolism, Pharmacokinetics, and Toxicity.

Despite extensive breakthroughs in chemistry, molecular biology, and genetics in the last decades, the success rates of drug development projects remain low. To improve predictions of clinical efficacy and safety of new compounds, a plethora of 3D culture methods of human cells have been developed in which the cultured cells retain physiologically and functionally relevant phenotypes for multiple weeks. Here, we critically review current paradigms for organotypic cultures of human liver, gut, and kidney such as perfused microchips, spheroids, and hollow fiber bioreactors and discuss their utility for understanding drug pharmacokinetics, metabolism, and toxicity. Furthermore, bioprinting and the microfluidic integration of different tissue models to mimic systemic drug effects are highlighted as promising technological trends. In the last part of the review, we discuss important considerations regarding the choice of culture substratum material to limit adverse effects such as drug absorption while facilitating the phenotypic maintenance of cultured cells. We conclude that recent advances in organotypic and microphysiological culture models of human tissues can improve drug development and contribute to an amelioration of clinical attrition rates. However, further validation, benchmarking, and consolidation efforts are needed to achieve more widespread dissemination and eventually regulatory acceptance of these novel tools.

Establishing Preferred Product Characterization for the Evaluation of RNA Vaccine Antigens.

The preferred product characteristics (for chemistry, control, and manufacture), in addition to safety and efficacy, are quintessential requirements for any successful therapeutic. Messenger RNA vaccines constitute a relatively new alternative to traditional vaccine development platforms, and thus there is less clarity regarding the criteria needed to ensure regulatory compliance and acceptance. Generally, to identify the ideal product characteristics, a series of assays needs to be developed, qualified and ultimately validated to determine the integrity, purity, stability, and reproducibility of a vaccine target. Here, using the available literature, we provide a summary of the array of biophysical and biochemical assays currently used in the field to characterize mRNA vaccine antigen candidates. Moreover, we review various in vitro functional cell-based assays that have been employed to facilitate the early assessment of the biological activity of these molecules, including the predictive immune response triggered in the host cell. Messenger RNA vaccines can be produced rapidly and at large scale, and thus will particularly benefit from well-defined and well-characterized assays ultimately to be used for in-process, release and stability-indications, which will allow equally rapid screening of immunogenicity, efficacy, and safety without the need to conduct often lengthy and costly in vivo experiments.

Comprehensive Evaluation of Organotypic and Microphysiological Liver Models for Prediction of Drug-Induced Liver Injury.

Drug-induced liver injury (DILI) is a major concern for the pharmaceutical industry and constitutes one of the most important reasons for the termination of promising drug development projects. Reliable prediction of DILI liability in preclinical stages is difficult, as current experimental model systems do not accurately reflect the molecular phenotype and functionality of the human liver. As a result, multiple drugs that passed preclinical safety evaluations failed due to liver toxicity in clinical trials or postmarketing stages in recent years. To improve the selection of molecules that are taken forward into the clinics, the development of more predictive in vitro systems that enable high-throughput screening of hepatotoxic liabilities and allow for investigative studies into DILI mechanisms has gained growing interest. Specifically, it became increasingly clear that the choice of cell types and culture method both constitute important parameters that affect the predictive power of test systems. In this review, we present current 3D culture paradigms for hepatotoxicity tests and critically evaluate their utility and performance for DILI prediction. In addition, we highlight possibilities of these emerging platforms for mechanistic evaluations of selected drug candidates and present current research directions towards the further improvement of preclinical liver safety tests. We conclude that organotypic and microphysiological liver systems have provided an important step towards more reliable DILI prediction. Furthermore, we expect that the increasing availability of comprehensive benchmarking studies will facilitate model dissemination that might eventually result in their regulatory acceptance.

Requirements for statistical safety demonstration for loss of coolant analyses

During the recent years, an increasing interest in computational safety analysis of Nuclear Power Plants (NPPs) is to replace the conservative evaluation model calculations by best estimate calculations supplemented by uncertainty analysis of the code results. Conservative means unfavorable in the direction of regulatory acceptance limits. The evaluation of the margin to acceptance criteria, e.g. the maximum fuel rod cladding temperature, should be based on the upper limit of the calculated uncertainty range. Uncertainty analysis is needed if useful conclusions are to be obtained from “best estimate” thermal-hydraulic code calculations, otherwise single values of unknown accuracy would be presented for comparison with regulatory acceptance limits. Methods have been developed and presented to quantify the uncertainty of computer code results. The Gesellschaft für Anlagen- und Reaktorsicherheit mbH (GRS) in Germany has proposed a statistical method, propagating input uncertainties by computer code calculations to end up with output uncertainties of the calculated results. The basic techniques of that GRS-method are presented for calculating the peak cladding temperatures of the fuel rods in a reactor core and to compare with an acceptance limit to be met under any design basis accident. A comparison of necessary number of code calculations is provided using a penalized “hot rod”, or a significantly higher number of calculations when all high powered fuel rods are considered in the analysis when conditions of many fuel rods are very close to the most unfavorable fuel rod.

Pig-a gene mutation database.

Mutations in the X-linked phosphatidylinositol glycan, class A gene (Pig-a) lead to loss of glycosylphosphatidylinositol (GPI) anchors and GPI-anchored proteins from the surface of erythrocytes and other mammalian cells. The Pig-a gene mutation assay quantifies in vivo gene mutation by immunofluorescent labeling and flow cytometry to detect the loss of GPI-anchored proteins on peripheral blood erythrocytes. As part of the regulatory acceptance of the assay, a public database has been created that provides detailed information on Pig-a gene mutation assays conducted in rats and mice. A searchable version of the database is available through a website designed and hosted by the University of Maryland School of Pharmacy. Currently, the database contains only mouse and rat data, but it is anticipated that it will expand to include data from other species, including humans. A major purpose in developing the database was to aid in the preparation of a Retrospective Performance Analysis and Detailed Review Paper required for Organisation for Economic Co-operation and Development Test Guideline acceptance. We anticipate, however, that it also will be useful for accessing and comparing Pig-a data to data from other assays and for conducting quantitative assessments of Pig-a gene mutation responses. Environ. Mol. Mutagen., 60:759-762, 2019. © 2019 Wiley Periodicals, Inc.

The challenges of delivering genetically modified crops with nutritional enhancement traits.

The potential for using genetic modification (GM) to enhance the nutritional composition of crops (for either direct human consumption or as animal feed) has been recognized since the dawn of the GM era, with such 'output' traits being considered as distinct, if not potentially superior, to 'input' traits such as herbicide tolerance and insect resistance. However, while input traits have successfully been used and now form the basis of GM agriculture, output trait GM crops are still lagging behind after 20 years. This is despite the demonstrable benefits that some nutritionally enhanced crops would bring and the proven value of GM technologies. This Review considers the present state of nutritional enhancement through GM, highlighting two high-profile examples of nutritional enhancement-Golden Rice and omega-3 fish oil crops-systematically evaluating the progress, problems and pitfalls associated with the development of these traits. This includes not just the underlying metabolic engineering, but also the requirements to demonstrate efficacy and field performance of the crops and consideration of regulatory, intellectual property and consumer acceptance issues.

Brazil Moves Toward the Replacement of Animal Experimentation.

In Brazil, efforts towards the regulatory acceptance and implementation of innovative methods to replace experimental animal use in various fields began to gather force in 2008, with the approval of Law No. 11,794/2008 (the Arouca Law). This law represented a milestone, as it created the National Council for the Control of Animal Experimentation (CONCEA) to deal with the ethical and legal issues related to the use of laboratory animals. In 2014, CONCEA put together a framework for expanding the implementation of non-animal methodologies for use in research and education. It also promoted the regulatory acceptance in Brazil of 24 test guidelines, including 15 in vitro approaches. It should be emphasised that, in Brazilian legislation, replacement is generally based on the toxicological endpoint and not on the category of product, as tends to be the case in other countries (e.g. cosmetics in the European Union). The resolution-dependent deadlines for the obligatory replacement of in vivo methods with the CONCEA-approved tests are 2019 and 2021. Brazil has advanced considerably towards the replacement of animal experimentation, and in certain aspects, this has been in a highly progressive manner. However, there is still a lot of work to be done, especially considering the current political scenario with reduced investment in research, development and innovation. The chronology of significant events following the approval of the Arouca Law, which have contributed to the promotion of the Three Rs alternatives in Brazil, will be examined.

From “Meat Culture” to “Cultured Meat”: Critically Evaluating the Contested Ontologies and Transformative Potential of Biofabricated Animal Material on Culture and Law

This article calls into question how different the parallel ontologies of meat are in terms of the realities they enact. It argues that while biofabricated animal material is expected to transform the production and consumption of meat, the material’s ontologies reinforce existing interrelationships between the dominant meat culture and the law. Additionally, the expectations surrounding biofabricated animal material vests the material with transformative power, even though it does not challenge the structural factors, namely law and culture, which underlie the exploitation and commodification of animals. I begin by delineating the co-constituting relationship between law, the realities of meat production and consumption, and meat culture. I then unpack the interface between different ontologies of meat and consider why the contest has taken place in legal terrains. From here, I consider the key expectations that have assigned transformative power to biofabricated animal material. I suggest that these expectations work to reinforce existing meat culture, and the configurations of power that benefit from it, as supported via legal instruments.

Workshop on the validation and regulatory acceptance of innovative 3R approaches in regulatory toxicology – Evolution versus revolution

At a joint workshop organized by RIVM and BfR, international experts from governmental institutes, regulatory agencies, industry, academia and animal welfare organizations discussed and provided recommendations for the development, validation and implementation of innovative 3R approaches in regulatory toxicology. In particular, an evolutionary improvement of our current approach of test method validation in the context of defined approaches or integrated testing strategies was discussed together with a revolutionary approach based on a comprehensive description of the physiological responses of the human body to chemical exposure and the subsequent definition of relevant and predictive in vitro, in chemico or in silico methods. A more comprehensive evaluation of biological relevance, scientific validity and regulatory purpose of new test methods and assessment strategies together with case studies that provide practical experience with new approaches were discussed as essential steps to build up the necessary confidence to facilitate regulatory acceptance.

Category consistency in the OECD QSAR Toolbox: Assessment and reporting tool to justify read-across

Read-across is an alternative or supplementary method to the use of test results (including test results from tests employing laboratory animals) for chemical hazard assessment. Its use by both the industrial and the regulatory communities has increased markedly over the past decade. A seminal issue for the regulatory acceptance of read-across predictions is reducing uncertainty in the prediction by evaluating similarity among the chemicals participating in the read-across. To assist in demonstrating and documenting this similarity, a “category consistency” functionality has been added to the OECD QSAR Toolbox. This module (semi-) automatically collects and displays information key to establishing physicochemical, structural and mechanistic similarities, as well as any experimental data associated with the endpoint under consideration. The Toolbox user can then employ this suite of information to draw a conclusion on the consistency of the recorded similarity measurements of the category members. The report writer provides documentation of the Toolbox information supporting the conclusion. The implementation of “category consistency” functionality is illustrated with a case example.

Determination of ceftriaxone in human plasma using liquid chromatography–tandem mass spectrometry

Ceftriaxone is a cephalosporin antibiotic drug used as first-line treatment for several bacterial diseases. Ceftriaxone belongs to the third generation of antibiotics and is available as an intramuscular or intravenous injection. Previously published pharmacokinetic studies have mainly used high-performance liquid chromatography coupled with ultraviolet detection (HPLC-UV) for the quantification of ceftriaxone. This study aimed to develop and validate a bioanalytical method for the quantification of ceftriaxone in human plasma using liquid chromatography followed by tandem mass spectrometry (LC-MS/MS). Sample preparation was performed by protein precipitation in combination with phospholipid-removal techniques for cleaning up matrix interferences. The chromatographic separation was performed on an Agilent Zorbax Eclipse Plus C18 column with 10 mM ammonium formate containing 2% formic acid: acetonitrile as mobile phase at a flow rate of 0.4 ml/min. Both the analyte and cefotaxime (internal standard) were quantified using the positive electrospray ionization (ESI) mode and selected reaction monitoring (SRM) for the precursor-product ion transitions m/z 555.0→396.1 for ceftriaxone and 456.0→324.0 for cefotaxime. The method was validated over the concentration range of 1.01-200 μg/ml. Calibration response showed good linearity (correlation coefficient > 0.99) and no significant matrix effects were observed. The intra-assay and inter-assay precision were less than 5% and 10%, respectively, and therefore well within standard regulatory acceptance criterion of ±15%.

High sensitivity methods to quantify chloroquine and its metabolite in human blood samples using LC-MS/MS.

Aim:Chloroquine is an antimalarial drug used in the treatment of Plasmodium vivax malaria. Three methods to quantify chloroquine and its metabolite in blood matrices were developed and validated.Methodology & results:Different high-throughput extraction techniques were used to recover the drugs from whole blood (50 μl), plasma (100 μl) and dried blood spots (15 μl as punched discs) followed by quantification with LC–MS/MS. The intra- and inter-batch precisions were below 15%, and thus meet regulatory acceptance criteria.Conclusion:The developed methods demonstrated satisfactory validation performance with high sensitivity and selectivity. The assays used simple and easy to automate extraction techniques. All methods were reliable with robust performance and demonstrated to be suitable to implement into high-throughput routine analysis of clinical pharmacokinetic samples.

Comparison of Toxicity-Weighted Disinfection Byproduct Concentrations in Potable Reuse Waters and Conventional Drinking Waters as a New Approach to Assessing the Quality of Advanced Treatment Train Waters.

Advanced treatment trains based on oxidation, biofiltration, and/or granular activated carbon (Ox/BAF/GAC) are an attractive alternative to those based on microfiltration, reverse osmosis, and advanced oxidation (MF/RO/AOP) for the potable reuse of municipal wastewater effluents, but their effluent quality is difficult to validate with respect to chemical contaminants. This study evaluated the sum of the concentrations of 46 disinfection byproducts (DBPs) after treatment by chlorine or chloramines weighted by metrics of toxic potency in 10 full- or pilot-scale reuse trains to estimate the DBP-associated toxicity of their effluents. These total toxicity-weighted DBP concentrations were compared to those measured in their local, conventional drinking waters as a benchmark for water quality receiving regulatory and widespread public acceptance. The results indicated that while the DBP-associated quality of MF/RO/AOP-based reuse waters can readily exceed that of drinking waters, that of Ox/BAF/GAC-based reuse waters can approach or exceed that of drinking waters, particularly when they are chloraminated. Unregulated, halogenated DBPs were the dominant contributors to the estimated DBP-associated toxicity. While RO/AOP treatment preferentially reduced the concentrations of the more toxic brominated DBP species, BAC and GAC treatment favored brominated DBP species by removing DOC but not bromide. Comparing the total toxicity-weighted DBP concentration between reuse and drinking waters provides drinking water as a rational benchmark for water quality comparison, explicitly recognizes that contaminants occur as mixtures, provides utilities flexibility in selecting the most efficient treatment trains to reduce estimated toxicity, and can be expanded to encompass new contaminants as toxic potency data become available.

Development, validation and application of a novel HPLC-MS/MS method for the measurement of minocycline in human plasma and urine

New treatments are urgently required to treat infections caused by multi-drug resistant Acinetobacter baumanni,. To address this need, a new formulation of Minocin®, (minocycline for injection) has been developed that allows for higher doses of minocycline to be administered. Phase 1 clinical trials were conducted in healthy volunteers to assess the safety and pharmacokinetics (PK) of this new formulation at higher doses. In order to generate PK data, novel, selective and simple HPLC-MS/MS based assays were developed and validated for the determination of minocycline (MC) in human plasma and urine. The respective working ranges were 0.05 to 30 mg/L and 0.1 to 30 mg/L. Removal of endogenous proteins with trichloroacetic acid was used as a simple means of extracting MC from the samples. An analogue, tetracycline was used as the internal standard (IS). Chromatographic separation, including that of MC from its 4-epimer (4-EMC), was achieved on a Waters XBridge BEH C18 column (50 x 4.6 mm ID, 5 μm) with gradient elution. The mobile phases comprised water containing 5 mM ammonium formate at a pH of 2.5, and methanol containing 5 mM ammonium formate. The internal standard (IS) was tetracycline, a structural analogue of minocycline.The methods were fully validated and met regulatory acceptance criteria for intra-run and inter-run accuracy and precision, carryover, dilution integrity and matrix effects. Mean extraction recoveries ranged between 64.3% and 84.6% for MC and 64.3% for the IS. There was no significant ion suppression or enhancement for MC or the IS. The validated assays were successfully applied to 1423 plasma and 689 urine samples from a Phase 1 clinical study.There was no evidence of instability, or significant interconversion between MC and 4-EMC, in stored clinical samples, spiked plasma and urine samples, or their extracts, under various test conditions.

Facilitators and Barriers for Implementing an Internet Clinic for the Treatment of Pressure Injuries.

Background:Pressure injuries (PIs) represent a frequent, often preventable, secondary complication of spinal cord injury (SCI) with serious consequences to health, societal participation, and quality of life. Specialized knowledge and service delivery related to treatment and prevention are typically located within major health centers.Introduction:For persons with SCI living at home, it can be challenging to access specialized PI care. A telehealth approach could help mitigate this challenge. This multisite pilot investigation assessed the feasibility of integrating information technologies within the management of PIs.Materials and Methods:Each study site formed a specialized interdisciplinary care team that identified components of their standard clinical care pathway and examined how they could be integrated with study technologies. A monitoring system was utilized to enable patients and caregivers to exchange clinical information with the care team.Results:Clinician and patient focus groups were completed to identify facilitators and barriers for long-term implementation. Findings demonstrate that this method of service delivery is feasible but requires further development.Discussion:This model of care requires refinement to address technological, regulatory, and clinician acceptance barriers; however, increased access to these services has the potential for improving PI healing or prevention rates in comparison with those not able to access specialized services.Conclusions:This project demonstrates that PI treatment services can be delivered effectively through the internet. Future trials can investigate efficacy and cost-effectiveness of this model of care to inform sustained implementation.