Ethical use of animals in medicine testing

The European Medicines Agency (EMA) supports the implementation of the so-called 3Rs principles - replace, reduce and refine - for the ethical use of animals in medicine testing across the European Union (EU). These principles encourage alternatives to the use of animals in the testing of medicines while safeguarding scientific quality and improving animal welfare where the use of animals cannot be avoided.

Updated on 24 March 2025:
'Scientific guidelines' section

Directive 2010/63/EU requires marketing authorisation holders to integrate the 3Rs and welfare standards for the treatment of animals in all aspects of the development, manufacture and testing of medicines.

The Directive aims to protect animals in scientific research, with the final aim of replacing all animal research with non-animal methods.

• Directive 2010/63/EU 

3Rs principles

The 3Rs stand for:

• replacing the use of animals with non-animal methods where possible;
• reducing the number of animals used to a minimum while still obtaining scientifically valid results;
• refining practices to minimise the stress and improve the welfare of study animals used for regulatory purposes.

 

3Rs visual: text version

Replace animal studies with non-animal methods

If this is not possible then:

Reduce animal studies to minimum required and necessary

In addition:

Refine practices to minimise stress of study animals

New approach methodologies

New approach methodologies (NAMs) refer to novel methods that are 3Rs-compliant. They may be incorporated in the assessment of the safety and efficacy of new medicines to replace or reduce animal use. Examples include in vitro (cell-based) systems and computer modelling.

These methods represent potential alternatives to animal testing in the non-clinical development phase of new medicines. Their use should be carefully considered.

Where animal testing is still needed, researchers should design and conduct such studies in line with the 3Rs principles.

EMA role

EMA supports the implementation of Directive 2010/63/EU and the 3Rs principles in the EU, by:

• identifying how the 3Rs can be implemented in regulatory testing;
• developing scientific guidelines for medicine developers and manufacturers;
• providing developers a forum for early dialogue via its Innovation Task Force (ITF) to discuss the regulatory acceptance of so-called new approach methodologies to replace the use of animals in in the testing of medicines;
• providing recommendations on 3Rs methods in the European Pharmacopoeia to help marketing authorisation holders comply with new or revised measures;
• carrying out a scientific review of batch release tests for human and veterinary vaccines and biologicals to ensure alignment with best practice in 3Rs;
• contributing to the development of harmonised guidance and requirements in Europe and globally, by working closely with relevant European and international bodies, such as the:
  
  • European Directorate for the Quality of Medicines and HealthCare (EDQM);
  • European Union Reference Laboratory for alternatives to animal testing (EURL-ECVAM);
  • International Council on Harmonisation of Technical Requirements for Registration of Pharmaceuticals for Human Use (ICH);
  • International Cooperation on Harmonisation of Technical Requirements for Registration of Veterinary Medicinal Products (VICH).

Working parties and other groups

EMA has a dedicated 3Rs Working Party (3RsWP), which provides advice to its scientific committees on all matters concerning the use of animals in regulatory testing of medicines.

EMA's Non-Clinical and New Approach Methodologies European Specialised Expert Community provides a forum for information-sharing between regulatory and academic experts on innovative non-clinical testing methods. These include new approach methodologies.

EMA has led on the establishment of an International Medicines Regulators' Working Group on 3Rs.

The group comprises members from EMA, the Swiss Agency for Therapeutic Products (Swissmedic), the Japanese Pharmaceuticals and Medical Devices Agency (PMDA), the Australian Therapeutic Goods Administration (TGA), Health Canada, and the United States Food and Drug Administration (FDA).

These agencies have agreed to work together to achieve the following:

• application of 3Rs in non-clinical testing;
• agreement on acceptance criteria for "New Approach Methodologies" (NAMs) within specific contexts of use;
• review of quality control and batch release requirements to encourage broader acceptance of the use of 3Rs-compliant methods where possible;
• support for the phasing out of obsolete tests;
• development of a regulatory position paper on 3Rs which could be shared with other medicines regulatory authorities (e.g., through The International Coalition of Medicines Regulatory Authorities (ICMRA));
• training and competence building through exchange of information on 3Rs-compliant methods (e.g. case studies, qualification approaches, acceptance criteria);
• sharing of information on 3Rs activities and developments in the participating regions, as well as opportunities for international stakeholder engagement (e.g. through scientific meetings/conferences, or through bodies responsible for validation, guidance development, and global regulatory coordination).

Terms of Reference (ToR) for the International Medicines Regulators’ Working Group on 3Rs

Reference Number: EMA/27380/2024

Long-standing commitment

The Joint Expert Group (JEG3Rs) and the 3Rs Working Group (3RsWG) preceded the establishment of the 3Rs Working Party, demonstrating EMA's long-standing commitment to the 3Rs.

For more information, see:

Statement of the European Medicines Agency position on the application of the 3Rs (replacement, reduction and refinement) in the regulatory testing of human and veterinary medicinal products

Reference Number: EMA/470807/2011Summary:

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.

EMA reporting on 3Rs

EMA published a report on the application of 3Rs in regulatory testing in 2016 and 2017 to support the implementation of the 3Rs principles.

Biennial report of the joint CVMP/CHMP working group on the application of the 3Rs in regulatory testing of medical products (2016/2017)

Adopted

EMA will resume its reporting on 3Rs activities and aims at issuing reports every two years.

This decision follows the establishment of the 3Rs Working Party in November 2022 and the end of business continuity measures in 2023. The latter dictated the suspension or reduction of the activities of non-product-related working parties / groups.

Scientific guidelines

EMA and its 3Rs Working Party (3RsWP) develop scientific guidelines to help medicine developers uphold the 3Rs principles and welfare standards for the treatment of animals in the testing of medicines. 

This is in line with comply with Directive 2010/63/EU.

Find relevant guidelines below:

• Guideline on regulatory acceptance of 3R (replacement, reduction, refinement) testing approaches
• Guidance for individual laboratories for transfer of quality control methods validated in collaborative trials with a view to implementing 3Rs

EMA regularly reviews and updates these guidelines to reflect developments in the field.

It also conducted a review of all EMA guidelines to ensure that they do not make reference to animal tests that are no longer considered appropriate. 

For more information and the guidelines that EMA has or will update as a result, see:

Review and update of EMA guidelines to implement best practice with regard to 3Rs (replacement, reduction and refinement) in regulatory testing of medicinal products – report on actions taken

AdoptedReference Number: EMA/CHMP/CVMP/3Rs/677407/2015

Reflection papers on 3Rs: public consultation

Two draft reflection papers on current regulatory testing requirements for both human and veterinary medicines are available for public consultation.

Find the draft documents via the links below: 

• Overview of the current regulatory testing requirements for medicinal products for human use and opportunities for implementation of the 3Rs
• Overview of the current regulatory testing requirements for veterinary medicinal products and opportunities for implementation of the 3Rs

The draft reflection papers incorporate new developments and approaches in the field of 3R.

The deadline to comment is 25 June 2025.

Other EMA positions and recommendations on 3Rs

Recommendations on 3Rs in European Pharmacopeia

EMA provides recommendations on 3Rs methods in the European Pharmacopoeia in the documents below. They help marketing authorisation holders comply with new or revised measures.

More information:

• European Pharmacopoeia Online

Recommendation to marketing authorisation holders, highlighting the need to ensure compliance with 3Rs methods described in the European Pharmacopoeia

AdoptedReference Number: EMA/CHMP/CVMP/JEG-3Rs/252137/2012Summary:

In accordance with the European Convention for the Protection of Vertebrate Animals Used for Experimental and Other Scientific Purposes, tests performed in animals must be carried out in such a way as to use the minimum number of animals and to cause the least pain, suffering, distress or lasting harm.

Recommendation to marketing authorisation holders, highlighting recent measures in the veterinary field to promote replacement, reduction, and refinement (3Rs) measures described in the European Pharmacopoeia - Applicabl...

AdoptedReference Number: EMA/CHMP/CVMP/3Rs/614768/2017Summary:

In accordance with the European Convention for the Protection of Vertebrate Animals Used for Experimental and Other Scientific Purposes (Council of Europe), tests performed in animals must be carried out in such a way as to use the minimum number of animals and to cause the least pain, suffering, distress or lasting harm. The European Pharmacopeia (Ph.Eur.) has, over the years, applied these principles by implementing alternative tests and assays that replace, reduce and refine animal use.

Recommendation to marketing authorisation holders, highlighting recent updates for reduction, refinement and replacement (3Rs) methods described in the European Pharmacopoeia applicable to human vaccines against hepatiti...

AdoptedReference Number: EMA/CHMP/CVMP/JEG-3Rs/243112/2015Summary:

In accordance with the European Convention for the Protection of Vertebrate Animals Used for Experimental and Other Scientific Purposes (Council of Europe), tests performed in animals must be carried out in such a way as to use the minimum number of animals and to cause the least pain, suffering, distress or lasting harm.

Recommendation to marketing authorisation holders, highlighting recent measures in the veterinary field to promote reduction, refinement and replacement (3Rs) measures described in the European Pharmacopoeia - Applicable...

AdoptedReference Number: EMA/CVMP/3Rs/336802/2017Summary:

In accordance with the European Convention for the Protection of Vertebrate Animals Used for Experimental and Other Scientific Purposes (Council of Europe), tests performed in animals must be carried out in such a way as to use the minimum number of animals and to cause the least pain, suffering, distress or lasting harm. The European Pharmacopeia (Ph. Eur.) has, over the years, applied these principles by implementing alternative tests and assays that reduce, refine and replace animal use.

Recommendation to marketing-authorisation holders for veterinary vaccines, highlighting the need to update marketing authorisations to remove the target animal batch safety test (TABST) following removal of the requireme...

AdoptedReference Number: EMA/CHMP/CVMP/JEG-3Rs/746429/2012Summary:

The requirement for the target animal batch safety test (TABST) for veterinary vaccines has been formally withdrawn from the European Pharmacopoeia, effective as of April 2013.

Veterinary medicine testing outside the EU

Wherever the manufacture or batch testing of veterinary medicines to be marketed in the European Economic Area (EEA) takes place, they must conform to EU ethical and animal welfare standards.

For more information, see:

Statement of the Committee for Medicinal Products for Veterinary Use position on the ethical use of animals in the testing, development and manufacture of veterinary medicines

Reference Number: EMA/CVMP/3Rs/506841/2017Summary:

Animal welfare is a value of the Union that is enshrined in Article 13 of the Treaty on the Functioning of the European Union (TFEU). The use of animals in scientific procedures including regulatory testing of veterinary medicinal products is strictly controlled within the European Union (EU), in accordance with Directive 2010/63/EU (hereinafter referred to as “the Directive”).