European Union Registration Research Articles

Safety and Immunogenicity of a Novel Influenza Subunit Vaccine Produced in Mammalian Cell Culture

Immunization remains the best prevention strategy for influenza, but production constraints for egg-based influenza vaccines have prompted the development of innovative cell culture manufacturing processes. Here, we describe a novel cell culture-derived influenza vaccine (CCIV) produced in Madin-Darby canine kidney cells. This phase 3, observer-blind, randomized, multicenter study in Poland compared the immunogenicity of a CCIV and a conventional egg-based vaccine. Participants, stratified by age (adults 18-60 years, n = 1300; elderly persons > or = 61 years, n = 1354), received a single intramuscular vaccination. Immunogenicity was assessed 21 days later by hemagglutination inhibition assay. Reactogenicity was assessed using self-completed diary cards. The immunogenicity of CCIV was noninferior to that of the conventional vaccine for all 3 vaccine strains in both age groups, regardless of underlying health status. Both vaccines fulfilled European Union registration criteria and were well tolerated, with similar incidences of solicited local and systemic reactions in both age groups; the only significant difference was an increased frequency of mild or moderate pain with CCIV than the conventional vaccine among adult (22% vs 17%; P < .05) and elderly (9% vs 5%; P < .001) vaccinees. CCIV was well tolerated and highly immunogenic in adults 18 years of age or older. Cell culture may offer greater flexibility of supply during periods of high demand for both seasonal and pandemic vaccines.

Data Sharing, Federal Rule of Evidence 702, and the Lions in the Undergrowth

Many concerns would find it useful to have a publicly available database recording exposures to particular substances, with contextual information. The European Union's Registration, Evaluations, Authorisation, and restriction of Chemicals regulation has increased interest in this. It has been suggested that journals should require detailed publication of exposure data with papers. But there are problems for researchers, for whom the data are a valuable resource which has been obtained with effort and often ingenuity. The publication could also raise problems of confidentiality and liability, and those who have to put the effort in publication are not those who benefit. Also, there are the problems of hostile critics misusing the information--for example, industry re-analysing data to counter any regulatory implications of a study--and this raises serious wider issues of editorial policy. Two books have recently given examples of industry misuse of science, 'Doubt is their product' by David Michaels and 'Defending the indefensible' by McCullogh and Tweedale. Michaels gives examples of hostile data reanalysis, and among other things, he discusses the impact on journals of US Federal Rule of Evidence 702, which encourages expert witnesses to try to get their testimony material into peer-reviewed journals. This certainly lies behind some submissions to this journal, and Michaels says that it has led to the creation of peer-reviewed journals which have strong industry influence. On the other hand, work funded by industry is not always wrong, papers from other sources are not always free from bias, and the problem for journals is to continue to apply consistent scientific standards in a sea of conflicting interests. It does not seem feasible or desirable for journals to insist on the publication of all the underlying data, although researchers might be encouraged to form consortia to share data.

QSAR models for reproductive toxicity and endocrine disruption in regulatory use – a preliminary investigation†

A special challenge in the new European Union chemicals legislation, Registration, Evaluation and Authorisation of Chemicals, will be the toxicological evaluation of chemicals for reproductive toxicity. Use of valid quantitative structure–activity relationships (QSARs) is a possibility under the new legislation. This article focuses on a screening exercise by use of our own and commercial QSAR models for identification of possible reproductive toxicants. Three QSAR models were used for reproductive toxicity for the endpoints teratogenic risk to humans (based on animal tests, clinical data and epidemiological human studies), dominant lethal effect in rodents (in vivo) and Drosophila melanogaster sex-linked recessive lethal effect. A structure set of 57,014 European Inventory of Existing Chemical Substances (EINECS) chemicals was screened. A total of 5240 EINECS chemicals, corresponding to 9.2%, were predicted as reproductive toxicants by one or more of the models. The chemicals predicted positive for reproductive toxicity will be submitted to the Danish Environmental Protection Agency as scientific input for a future updated advisory classification list with advisory classifications for concern for humans owing to possible developmental toxic effects: Xn (Harmful) and R63 (Possible risk of harm to the unborn child). The chemicals were also screened in three models for endocrine disruption. †Presented at the 13th International Workshop on QSARs in the Environmental Sciences (QSAR 2008), 8–12 June 2008, Syracuse, USA.

Variable Access to Clopidogrel in a Harmonized EU Market

Objectives This study focuses on the different national coverage and reimbursement strategies and their consequences for access to clopidogrel, a drug with a central European Union (EU) registration. Our objectives are 1) to assess whether changes in reimbursement policies in EU member states influenced clopidogrel prescribing; and 2) to determine whether clopidogrel-specific policy characteristics, general characteristics of the health system, or indicators for the amount of cardiovascular care delivered were associated with the level of clopidogrel prescribing. Methods Data were collected in Austria, Belgium, Denmark, Germany, Hungary, Portugal, Slovenia, The Netherlands, and the United Kingdom (England). Utilization rates were expressed as defined daily doses (DDDs)/1000 persons/day. To determine whether changes in reimbursement policies influenced clopidogrel utilization, a segmented linear regression approach was used. Results Clopidogrel prescribing varied widely in the studied countries, from 2.76 (The Netherlands) to 6.83 (Belgium) DDDs/1000 persons/day (March 2005). Six countries had therapeutic indication restrictions to clopidogrel use. Health system characteristics did not explain variation in clopidogrel prescribing. Conclusion A disconnect will be indicated in this study between the concept of a harmonized EU pharmaceuticals market and the reality in an individual member state. Although clopidogrel was centrally registered in the EU, policy measures at the national level result in different roles in clinical practice for this drug.

A new approach to the hazard classification of alloys based on transformation/dissolution

Most of the metals produced for commercial application enter into service as alloys which, together with metals and all other chemicals in commerce, are subject to a hazard identification and classification initiative now being implemented in a number of jurisdictions worldwide, including the European Union Registration, Evaluation, Authorisation and Restriction of Chemicals (REACH) initiative, effective 1 June 2007. This initiative has considerable implications for environmental protection and market access. While a method for the hazard identification and classification of metals is available in the recently developed United Nations (UN) guidance document on the Globally Harmonized System of Hazard Classification and Labelling (GHS), an approach for alloys has yet to be formulated. Within the GHS, a transformation/dissolution protocol (T/ DP) for metals and sparingly soluble metal compounds is provided as a standard laboratory method for measuring the rate and extent of the release of metals into aqueous media from metal-bearing substances. By comparison with ecotoxicity reference data, T/D data can be used to derive UN GHS classification proposals. In this study we applied the T/DP for the 1st time to several economically important metals and alloys: iron powder, nickel powder, copper powder, and the alloys Fe-2Cu-0.6C (copper = 2%, carbon = 0.6%), Fe-2Ni-0.6C, Stainless Steel 304, Monel, brass, Inconel, and nickel-silver. The iron and copper powders and the iron and nickel powders had been sintered to produce the Fe-2Me-0.6C (Me = copper or nickel) alloys which made them essentially resistant to reaction with the aqueous media, so they would not classify under the GHS, although their component copper and nickel metal powders would. Forming a protective passivating film, chromium in the Stainless Steel 304 and Inconel alloys protected them from reaction with the aqueous media, so that their metal releases were minimal and would not result in GHS classification. For the other alloys, we developed a new critical surface area-toxic units (CSA-TU) approach to derive their GHS classification proposals. The CSA-TU approach can be readily applied to other multicomponent alloy systems, without the need to arbitrarily select a particular component among several as the determinant of toxicity. This paper shows how regulatory obligations, such as those mandated by REACH, can be met with a laboratory-based CSA-TU method for deriving hazard classification proposals for alloys, linking to attendant environmental protection management decisions. Drawing on T/D data derived from laboratory testing of the alloy itself, the CSA-TU approach can be applied to establish scientifically defensible decisions on hazard classification proposals for an alloy of interest. The resulting decisions can then be incorporated into environmental management measures in such jurisdictions as the European Union. Based on an approach developed specifically for alloys, the hazard classification decisions can be regarded as relevant, credible, and protective of the environment. Since alloys are usually more resistant to chemical attack than their components, this approach is a considerable improvement over the possibility provided for in the GHS of calculating a hazard classification level for an alloy from the classification levels of its components.

Bioaccumulation Factors for PCBs Revisited

Bioaccumulation factors (BAFs) for individual polychlorinated biphenyl (PCB) congeners in Barents Sea and White Sea marine calanoid copepods were 1-3 orders of magnitude higher than BAFs in the same species in Canadian and Alaskan Arctic Ocean areas, and in freshwater plankton (Lake Ontario) reported from the mid- to early 1980s. The present study reviews variability in PCB BAFs from the North American Great Lakes and the Arctic Ocean, and discusses possible explanations for the large variation among different studies. BAFs are higher in recent arctic marine and Great Lakes studies than previously reported, and they are at least 10 times higher than those predicted from the octanol-water partition coefficient (KOW). If the recent high BAFs are realistic, it means that earlier reported BAFs are too low. This is likely due to earlier erroneously high quantification of water PCB concentrations, and it implies that bioaccumulation in zooplankton is more efficient than previously assumed. Evidence is presented supporting that also trophic transfer and biomagnification of PCBs in zooplankton leads to BAFs well above those predicted by simple equilibrium partitioning. Overall, miss-measurement of water PCB concentrations and biomagnification contribute significantly to variability in BAFs for PCBs within and among studies. This large variability of BAFs for PCBs in zooplankton illustrated in the present study is of importance for future assessments of potential new bioaccumulative chemicals that rely on measured BAFs, such as the European Union Registration, Evaluation and Authorization of Chemicals program (REACH).

The Case for Taking Account of Metabolism when Testing for Potential Endocrine Disruptors In Vitro

Legislation in the USA, Europe and Japan will require that chemicals are tested for their ability to disrupt the hormonal systems of mammals. Such chemicals are known as endocrine disruptors (EDs), and will require extensive testing as part of the new European Union Registration, Evaluation and Authorisation of Chemicals (REACH) system for the risk assessment of chemicals. Both in vivo and in vitro tests are proposed for this purpose, and there has been much discussion and action concerning the development and validation of such tests. However, to date, little interest has been shown in incorporating metabolism into in vitro tests for EDs, in sharp contrast to other areas of toxicity testing, such as genotoxicity, and, ironically, such in vitro tests are criticised for not modelling in vivo metabolism. This is despite the existence of much information showing that endogenous and exogenous steroids are extensively metabolised by Phase I and Phase II enzymes both in the liver and in hormonally active tissues. Such metabolism can lead to the activation or detoxification of steroids and EDs. The absence of metabolism from these tests could give rise to false-positive data (due to lack of detoxification) or false-negative data (lack of activation). This paper aims to explain why in vitro assays for EDs should incorporate mammalian metabolising systems. The background to ED testing, the test methods available, and the role of mammalian metabolism in the activation and detoxification of both endogenous and exogenous steroids, are described. The available types of metabolising systems are compared, and the potential problems in incorporating metabolising systems into in vitro tests for EDs, and how these might be overcome, are discussed. It is recommended that there should be: a) an assessment of the intrinsic metabolising capacity of cell systems used in tests for EDs; b) an investigation into the relevance of using the prostaglandin H synthase system for metabolising EDs; and c) a feasibility study into the generation of genetically engineered mammalian cell lines expressing specific metabolising enzymes, which could also be used to detect EDs.

The Regulatory Acceptance of Alternatives in the European Union

Recently, progress has been made toward the regulatory acceptance of replacements in the European Union (EU), particularly with the introduction of in vitro methods for the prediction of skin corrosivity, dermal penetration, phototoxicity and embryotoxicity. In vitro genotoxicity tests are well established, and testing for this endpoint can be completed without animals, provided that clear negative outcomes are obtained. Tiered approaches including in vitro tests can also be used to address skin and eye irritation endpoints. Reductions and/or refinements in animal use are being achieved following the replacement of the oral LD50 test with alternative methods and the adoption of reduced test packages for materials, such as closed-system intermediates and certain polymers. Furthermore, the use of a "read-across" approach has reduced animal testing. Substantial gains in refinement will also be made with the recent acceptance of the local lymph node assay for skin sensitisation and the development of an acute inhalation toxicity method that avoids lethality as the endpoint. For the future, under the proposed EU Registration, Evaluation and Authorisation of Chemicals (REACH) scheme, it is envisaged that, where suitable in vitro methods exist, these should be used to support registration of substances produced at up to ten tonnes per annum. This proposal can only accelerate the further development, validation and regulatory acceptance of such alternative methods.

Ten Years of Experience with the Trivalent Split-Influenza Vaccine, Fluarix???

The trivalent inactivated split-influenza vaccine, Fluarix™, has been available since 1992. To assess the safety and immunogenicity of the vaccine from studies in healthy adults and the elderly performed from 1992 to 2001 to comply with European Union registration requirements, and from studies in children and populations at high risk of severe influenza illness or influenza-related complications. Annual registration studies were performed in healthy adults aged 18–60 years and the elderly aged >60 years. Six studies in children aged 6 months-18 years, and five studies in adults with high-risk conditions, were done during the same period. The high-risk populations included immunosuppressed patients with solid tumours or systemic malignancies, renal or liver transplant recipients, and patients with chronic obstructive pulmonary disease or insulin-dependent diabetes mellitus. Adults and children aged >36 months received 0.5ml of the vaccine by intramuscular or deep subcutaneous injection. Children aged 6–35 months received 0.25ml; in some children not previously vaccinated or exposed to influenza, a second dose was given 4 weeks later. Immunogenicity was determined by measuring haemagglutination inhibiting antibodies just before and 21 ± 2 days after vaccination. Redness, swelling, induration and pain at the injection site were recorded, along with systemic symptoms such as fever. In all registration studies (n = 1 558), the vaccine consistently exceeded European Union immunogenicity criteria. Geometric mean titres in adults peaked 21 days after vaccination and remained above the protection level for all three strains for up to 12 months. In healthy adults and the elderly, seroprotection rates were 69–100% and consistently exceeded 82% from 1996. Immunogenicity in children (n = 364) and high-risk populations (n = 285) exceeded the targets for healthy adults. The vaccine was well tolerated in all age groups and populations. The most frequently reported symptoms in healthy adults and the elderly were local redness of >20mm (10–31%) and swelling of >20mm (11–16%). The incidence of fever >38.5°C was 37°C was also rare (0–5%) in children over 6 years old, but was more frequent in younger children (11–27%). The reactogenicity of the vaccine was low in the high-risk populations. Fluarix™ has a good safety profile and is highly immunogenic in all age groups as well as in high-risk populations. The all-year persistence of antibodies means that the vaccine gives protection for the entire influenza season.