Tải bản đầy đủ - 0trang
2 Animal Reduction Opportunities in Genotoxicity In Vivo Testing: Integration in Standard Toxicity Studies and Combination of Endpoints
Regulatory Acceptance of Alternative Methods in the Development…
investigated the sensitivity and practicality of integrating the liver Comet assay into
acute and 2- or 4-week repeat-dose rat toxicity studies (Rothfuss et al. 2010). These
and other data were considered by the Basle 2009 IGWT working groups as part of
their recommendations on in vivo genotoxicity testing, in particular the suitability
of tests with multiple genotoxicity endpoints integrated into acute or RDT studies.
The combination of the acute in vivo MN and Comet assays was considered by the
working group to represent a technically feasible and scientiﬁcally acceptable alternative to conducting independent assays. For the integration of Comet assays into
RDT studies, the working group reached the consensus that, based upon the limited
amount of data available, integration is scientiﬁcally acceptable and that the liver
Comet assay can complement the MN assay in blood or bone marrow in detecting
in vivo genotoxins. Practical issues need to be considered when conducting an integrated Comet assay study (Rothfuss et al. 2010).
The ICH EWG has adopted these advices for combining different endpoints into
one study in the revised ICH S2 guideline. When in vivo assessment of genotoxicity
with two tissues is required the guideline encourages the incorporation of two genotoxicity assays in one study using the same animals, e.g. bone marrow micronucleus
test and liver DNA strand breakage assay.
Recent study report submissions to regulatory health authorities clearly indicate
that the new options offered by the ICH S2R1 are increasingly utilized by pharmaceutical industry. In particular, integration of MN analysis in RDT studies in cases
where in vitro testing is negative and acute combined in vivo MN/comet assay to
follow-up positive ﬁndings in in vitro mammalian cell tests are the preferred options
in recently performed genotoxicity testing programs. It can thus be roughly estimated that the number of animals used for in vivo genotoxicity assessment in new
drug development may be decreased by nearly 50 % as a result of the revisions of
the ICH S2 guideline.
In summary, the experiences with the process leading to revision of the ICH S2
guideline clearly show that formal validation is not a necessary prerequisite for
regulatory acceptance of new 3R testing approaches. Instead, the scientiﬁc credibility of new methodologies and its use for regulatory purposes is pragmatically
assessed in a formalized ICH procedure by a working group of recognized industry
and regulatory experts in the ﬁeld based on high quality data from different sources.
The EMA Approach to Regulatory Acceptance
of 3R Test Methods
The JEG 3Rs
To demonstrate its commitment to the application of replacement, reduction and
reﬁnement (the 3Rs) of animal testing as detailed in Directive 2010/63/EU (Directive
2010), in October 2010, the EMA endorsed the establishment of a Joint ad hoc
Expert Group on the application of 3Rs in the development of medicinal products
S. Beken et al.
(JEG 3Rs). The JEG 3Rs has as a mandate to improve and foster the application of
3Rs in the regulatory testing of medicinal products throughout their lifecycle.
Moreover, this group provides advice and recommendations to the Committees (i.e.
CHMP and CVMP) on all matters related to the use of animals in regulatory testing
of medicinal products.
The core of the JEG 3Rs consists of experts from CVMP and CHMP and their
working parties for which animal testing is relevant, and can be complemented, as
necessary, by speciﬁc experts. As such, all relevant disciplines (i.e. quality, safety
and, in the case of veterinary medicinal products, efﬁcacy) are represented, for both
pharmaceutical and biological/immunological products.
As it is recognised that much work is already being done in the 3Rs area by other
European Commission bodies and consequently it is considered that duplication of
efforts should be avoided, the JEG 3Rs works in close cooperation with EURL
ECVAM (European Union Reference Laboratory for Alternatives to Animal
Testing) and EDQM (European Directorate for the Quality of Medicines and
Amongst the achievements of the JEG 3Rs so far are the efforts to ensure compliance of existing regulatory guidance with the 3Rs. As such, a concept paper
announcing a review of existing EMA guidance to ensure compliance with best 3Rs
practice was published in February 2014 (Concept paper on review and update of
European Medicines Agency Guidelines 2014) and publication of the ﬁrst amended
guidelines will ensue. Eight guidelines have so far been identiﬁed for revision but
the review is not yet complete. In addition, a draft guideline on regulatory acceptance of 3Rs testing approaches has been published for a 3-month period of consultation (Draft Guideline on regulatory acceptance of 3R 2014). This guideline
describes the process for submission and evaluation of a proposal for regulatory
acceptance of 3R (Replacement, Reduction and Reﬁnement) testing approaches for
use in the development and quality control during production of human and veterinary medicinal products. Furthermore, scientiﬁc and technical criteria for validation
of 3R testing approaches are presented and pathways for regulatory acceptance of
3R testing approaches are described.
Another work stream of the JEG 3Rs is related to the implementation of the 3Rs
in batch release testing of human and veterinary medicinal products. To this end, a
statement was published highlighting the need for marketing authorization holders
(MAHs) to ensure that batch safety and potency tests comply with 3Rs options
available in the Ph Eur (Recommendation to marketing authorisation holders 2012).
A statement was published highlighting the need for MAHs for veterinary vaccines
to update MAs to remove the target animal batch safety test following removal of
the requirement from the Ph Eur (“Recommendation to marketing authorisation
holders” 2013). At their July 2014 meetings CXMP adopted a concept paper for
publication for consultation, announcing the development of guidance on transferring validated quality control methods to a product/laboratory speciﬁc context
(Concept paper on tranferring quality control methods 2014). Work on developing
the guideline is now underway. In addition, review of batch release testing for
human and veterinary vaccines is conducted and follow-up communications to
Regulatory Acceptance of Alternative Methods in the Development…
JEG 3Rs coordinates responses to requests from the European Union Reference
Laboratory for Alternatives to Animal Testing (EURL ECVAM) for preliminary
analysis of regulatory relevance of new alternative methods (PARERE).
The existence of the JEG 3Rs provides a strong signal indicating that EMA takes
3Rs issues seriously. A clear statement of the EMA on the application of the 3Rs in
regulatory testing of human and veterinary medicinal products was published in the
EMA website at the very beginning of the JEG 3Rs’ ﬁrst mandate (Statement of the
EMA position on the application of the 3Rs 2011). This reads as follows:
The European Medicines Agency (EMA) commits to the application of replacement, reduction and refinement (the 3Rs) of animal testing as detailed in Directive
2010/63/EU1. To this end, a Joint ad hoc Expert Group (the JEG 3Rs) has been
created in order to promote best practice in the implementation of the 3Rs in regulatory testing of medicinal products and to facilitate full and active cooperation with
other European groups working in the 3Rs area.
While significant progress has been made in relation to regulatory testing involving animals it remains the case that certain types of data can only be generated by
means of animal studies. Where such studies are needed they should be selected and
conducted in strict adherence to the 3Rs principles.
As a European body with responsibility for developing harmonised European
regulatory requirements for human and veterinary medicinal products the EMA has
and will continue to play a key role in eliminating repetitious and unnecessary animal testing in the European Economic Area (EEA), in collaboration with other
European organisations such as EDQM. Through its active participation and collaboration in the work of other multinational organisations such as the ICH and the
VICH, the EMA contributes to the application of the 3Rs in the development of
globally harmonised requirements, the implementation of which contributes to the
elimination of unnecessary animal testing.
The JEG 3Rs is now recognised at international level and is often cited as an
example of how regulatory agencies should tackle 3Rs issues whilst providing a
clear entry point for questions or comments in this area. The JEG 3Rs mandate was
renewed in October 2014 for the second time, thus allowing this group to continue
its work in order to achieve progress in the ﬁeld of 3Rs, an area for which Europe is
clearly a global frontrunner.
Draft Guideline on Regulatory Acceptance of 3R
The application of the 3Rs were already highlighted in the Position on the
Replacement of Animal studies by in vitro models (Replacment of animal studies
by in vitro models 1997), adopted by the then called EMA Committee on Proprietary
Medicinal Products (CPMP) at its meeting in February 1997. This Position Paper
S. Beken et al.
addressed the feasibility of replacing in vivo animal studies by in vitro investigations in the preclinical development of medicinal products. In addition, considerations regarding validation procedures for in vitro methods and their incorporation
into CPMP Notes for Guidance were presented.
Whilst replacement of animal studies remains the ultimate goal, the application
of all 3Rs needs to be the focus. As exempliﬁed by the ICH regulatory safety guidelines described earlier, approaches aiming at reducing or reﬁning animal studies are
being routinely implemented in regulatory guidelines, where applicable. At the
same time, over the past years, new in vitro methods have been accepted for regulatory use via multiple and ﬂexible approaches, either as pivotal, supportive or as
exploratory mechanistic studies, wherever applicable. As such, although regulatory
acceptance of 3Rs testing approaches is currently possible, a formal regulatory
acceptance process has been lacking and implementation of new test methods in
routine regulatory testing has sometimes proven problematic.
Consequently, a review of the position paper focusing primarily on Replacement
was needed. As such, on March 11th 2011 a Concept Paper on the Need for Revision
of the Position on the Replacement of Animal studies by in vitro models (Concept
paper on the Need for Revision of the Position on the Replacement of Animal
Studies 2011) was drafted by the CHMP Safety Working Party and published on the
EMA website. Herein, aside from the extended focus to all 3Rs principles, the revision intended to describe a clear process for regulatory acceptance of 3Rs testing
approaches, to discuss qualiﬁcation criteria and bring the requirements in line with
As 3Rs principles apply to all regulatory testing requirements involving animal
use for both human and veterinary medicinal products, a multidisciplinary drafting
group was set up under the JEG 3Rs to develop the draft Guideline for Regulatory
acceptance of 3Rs testing approaches (Draft Guideline on regulatory acceptance of
3R testing approaches 2014). Concomitantly the JEG 3Rs started a thorough review
of the current regulatory testing requirements for human and veterinary medicinal
products and identiﬁcation of opportunities for implementation of the 3Rs.
The Draft guideline was forwarded to the relevant EMA Working Parties and
Committees for comments on the 17th of March 2014 and the ﬁnal draft was
launched for public consultation on the 3rd of October 2014. Comments received
are being considered and an update of the guideline is currently under way.
Draft Guideline for Regulatory Acceptance of 3Rs Testing
This guideline only applies to testing approaches that are subject to regulatory guidance for human and veterinary medicinal products. More speciﬁcally, those that are
used to support regulatory applications, such as clinical trial and marketing authorisation applications. The process of uptake of 3Rs testing methods in the Ph. Eur.
Monographs is excluded.