- Presenter
Welcome to EDQM On Air, a podcast on public health brought to you by the European Directorate for the Quality of Medicines and Healthcare of the Council of Europe. We hope you enjoy this episode and we invite you to stay tuned to learn how the EDQM and its stakeholders work together for better health for all. One of the EDQM's latest and most proud achievements is the phasing out of the rabbit pyrogen test, or RPT, from the European Pharmacopoeia, an effort which is now in its final stages of implementation. After being introduced into the European Pharmacopoeia in the 1970s, the RPT has been widely used in testing ever since. However, it sadly leads to suffering and often death for the animals that are being subjected to these tests. Fortunately, ever since its introduction, viable alternatives have been discovered and refined. meaning that it is no longer deemed necessary to continue its use. The EDQM, in close collaboration with its stakeholders, its partner organizations, and its numerous groups of experts, has been on the front line of this effort. In June 2024, the removal of the RPT from the European Pharmacopeia finally became a reality, after the official adoption of new texts that came to replace it. But how did this all take place? Who were the people that spearheaded this undertaking, and how did they achieve it? To help us better understand this milestone development, Gwenaël Cirefice and Emmanuelle Charton from the EDQM kindly agreed to come on the podcast as guests. We'd like to thank you for tuning in, and we hope this episode gives you a clearer idea about our commitment to animal welfare in Europe and beyond. All right. So, Gwenaël, Emmanuelle, thank you for coming on the podcast.
- Emmanuelle Charton
Hello.
- Presenter
Hello once again so I'll get straight into it. Today as I said in the intro we're going to talk about the rabbit pyrogen test so could you tell me a bit about what it is exactly and when did it start being used in pharmaceutical testing and perhaps a bit about the legal framework around it?
- Gwenaël Cirefice
Yeah so the the rabbit pyrogen test so is a test that is performed on rabbits And it's intended to check if medicines contain certain contaminants that cause fever reaction in humans. And these contaminants, we call them pyrogen. And this is a test that has been used by manufacturers of injectable medicines. It's a historical test that was introduced in the European Pharmacopoeia in the 70s. And then the principle of the test is, so you would inject the medicine into rabbits, and if they develop a fever, the contaminants are considered to be present. So it's a safety test. And the same rabbits are used repeatedly in the test, which causes suffering, and also eventually the death of the animal. It's a test, to give you an idea, it's a test that was reported. to consume approximately 25,000 rabbits in the EU alone, and around 400,000 rabbits worldwide, globally. Maybe for those who are unfamiliar with it, I think it's good to explain the concept of 3Rs, so in the context of alternatives to animal testing. So concept of 3Rs refers to replace, that's the first R, reduce and refine the use of animals that are used in scientific experiments. So you replace an animal test by a non-animal test and as a result no animals are used anymore in the experiment. Reduce, so you reduce the animal use to the minimum required or minimum necessary. As a result, fewer animals are used to achieve the aim of the test. And then refine is to refine the test to minimize the stress to the animals. So you would use a test that causes less suffering, less distress to the animals. And I think what is important to stress is that EDQM as an organization has been a leading force in the application of this concept of three R's to the quality control of medicines and with many achievements over the years. There is also a specific legal framework in Europe regarding animal testing and animal welfare. Number one is a convention of the Council of Europe. So it's called the European Convention for the Protection of Vertebrate Animals that are used for experimental and other scientific purposes. And then number two is the EU Directive 2010-63 EU. It's on the protection of animals used for scientific purposes. The first, so the convention, so it's a treaty that encourages not to experiment on animals, except of course if there's no other choice. It gives you a number of principles on how to improve animal welfare. And then the second, the EU directive regulates the use of animals that are used for experiments. And I think maybe the key principles to remind is that this requires the use of non-animal methods whenever they are available. Right.
- Presenter
So is there no alternative methods?
- Gwenaël Cirefice
Yes, so there are indeed alternatives to the rapid biogen test, so tests that are not using animals. One of these tests is called the monocyte activation test, or MAT. So that is a test that is using human cells or white blood cells. And it mimics the fever reaction that happens in humans when there are pyrogens, but in the test tube. And then a second such test is called the bacterial endotoxin test or BET. So it's a test that detects components of the cell walls of bacteria. which are called endotoxins. And in the presence of endotoxin, there will be a reaction, essentially formation of a clot or development of a color.
- Presenter
So it does the same job, basically, as the rabbit pyrogen test?
- Gwenaël Cirefice
So the first test, MAT, is able to detect all pyrogens, so it does the same job as the rabbit pyrogen test. Now the second test, the BET, detects only certain types of pyrogens, but which are the more common pyrogens. which we call the endotoxins. All right.
- Presenter
So there is alternatives. And from what you just said, there was also an effort towards potentially phasing it out or reducing it, or at least creating a framework around it to control it, right, the testing on rabbits. So now the rabbit pyrogen test is very close to being actually concretely phased out. So what are the key events that led to this development?
- Emmanuelle Charton
Yes, exactly. So you had to imagine the situation that Gwenaël just described. So European Pharmacopeia with a chapter describing the rabbit pyrogen test, some other chapters that do not use animals, the legal framework that encourages the phasing out of using of live animals in laboratories, and on top of that, about 60 texts of the Pharmacopeia. which referred to the rabbit pyrogen test. But there is one detail which is important that I want to highlight, is that in the chapter describing the rabbit pyrogen test of the pharmacopeia, you have the protocol. So the protocol is that you inject the medicine to the ear of the rabbit, and you measure the rise of temperature in the rabbit. But at the end of the chapter, there is a paragraph which says, in the... interest of animal welfare, you should validate in vitro method instead of the animal method. In vitro means a method which is used in a laboratory without using an animal. And so as far as we were concerned at EDQM, we thought this is enough. There is this paragraph which says, do not use the animals. But in fact, it was not because in Europe, rabbits continue to be widely used. For all these 60 texts described in the Pharmacopeia, going from antibiotics to vaccines, also radiopharmaceuticals, blood products, they were widely used. And then the trigger was, in fact, some external messages we got from our collaborators. And I would like to cite a few names here. I think it's important because we had, for example, Uwe Liebke from Germany. He's an antibiotic specialist. We had Christina von Hünnolstein from Italy, who was one of our experts in the vaccine field. We had Peter Turacek from Austria in the blood field and also very important. Important person who played an extremely important role in this whole exercise, Ingo Spreitzer from Germany. And I personally got to know Ingo more than 20 years ago when he was doing his PhD on the MIT. So these...
- Presenter
The test we mentioned before.
- Emmanuelle Charton
Exactly. Yes. So this was a group of protagonists, I would say, from our connection with experts. Now, there was another trigger, which was the... Collaboration between EU and the industry is called EPAA, the European Partnership for the Approach Using Alternatives to Animals. And this collaboration, they approached us many times in the last years with some proposals to remove the Pyrogen test. And it was Blaise Descampes in 2020 who approached us with a list of our texts with proposals to remove the Pyrogen test. So together, Blaise and all the other experts I cited, we decided to put them all together because they had the motivation and the enthusiasm to get rid of the rabid Parisian test. So in 2020, in the middle of the pandemic, we put all these persons together, asking them to find a strategy to tackle all these 60 texts at the same time. Because in the years before we had... a request here and there for one or two of the texts, and then a few years after for another text. And we thought now is the time to address that for all the 60 texts at once. And these people, they designed a strategy. It took one year and the summer of 2021, the strategy was finalized and the European Pharmacopeia Commission decided to embark on this strategy and it was largely publicized. And we announced a five-year project. So at the time, our expert, Christina, said, oh, but I will be retired after the five years. But in the end, the five years went very rapidly. And what is very important to say is at the time, it was communicated. We said to our public, please be aware, prepare yourself. In five years, you will not be able to use a rabbit anymore to measure. fever-inducing substances in pharmaceutical products. And we made it because in fact, we went to conferences, we seized every opportunity of discussions in scientific congresses to advertise the fact that we were going to do that. And it was very interesting to see that a few months already after this announcement, we had companies who had either decided to develop kits for MAT tests, or also if they were producers of these pharmaceutical products, they had installed their own planning on how to phase out the rabbit in their own facilities, announcing we are ready for 2026. So it took all those years to, I would say, put in practice the strategy for phasing out. So communication was a very important factor to that.
- Presenter
Okay, so there was a strategy put in place, there were the people that were willing to be involved, and from the way you explained it, it looks like everything was set for the process to begin. So could you go a bit more in depth about how it actually was achieved? What were the first concrete steps that were taken to actually go ahead with the phasing out of rabbit pyrogen testing?
- Gwenaël Cirefice
That's a good question. So as you heard, there were about 60 European Pharmacopoeia texts, and they were covering diverse topics, as you heard. 60 texts that required to perform the rapid collagen test, and therefore these 60 texts had to be examined by a group of experts as part of this exercise. So the strategy that Emmanuelle has just mentioned to replace the rapid collagen test is... based on two elements. So of course, the deletion of the rapid pyrogen test formed the 6D text. But a key element, the second part, is the elaboration of a new chapter of the pharmacopoeia called Pyrogenicity. And this new chapter gives guidance to the users on how to select and implement a suitable test not using animals. to control the biogenicity of their products. So this new general chapter requires the users to carry out a risk assessment, and based on the risk assessment they would select suitable non-animal tests to control the pyrogenicity. Okay,
- Presenter
so from one hand you remove the possibility of doing the rabid pyrogen test because it's not in the pharmacopoeia anymore, and then you install more robust alternatives to it.
- Gwenaël Cirefice
Yes, precisely, with the tools, with the non-animal tools to control the power chain content in these products. Okay. Emmanuelle mentioned the communication part, very important for our project. Another part was collaboration, was absolutely essential. The project required collaboration between 11 groups of experts of the European Pharmacopeia, so in various fields. In total, there were more than 250 experts who took part to this whole exercise. One of the groups, which we call the BET Working Party, so it was the group with the technical expertise on the techniques, so MAT, BET, played a central role in the project. They were in charge of elaborating the new chapter. And then regarding the collaboration across the groups, the feedback we received from the consultation of the individual group of experts served to really construct, to enrich the overall strategy on phasing out the rapid biogen test. There was also the chair of the Beating Working Party, Ingo Schpreitzer, that Emmanuelle mentioned, who personally attended most of the discussions in the plenary meetings of the 10 other group of experts to really provide his technical input into the discussion. and help into building this strategy. So there was really synergies, interactions across the 11 groups of experts that were part of this exercise. There was also involvement of many colleagues within the organization, within EDQM, around 50 colleagues in total, who were involved to prepare, of course, 60 texts. That's a massive number. So scientific administrators, assistants, so these are the people who are supporting the group of experts of the Pharmacopoeia. Scientific editors, proofreaders, translators, and our publication team. And I think a nice anecdote is that at some point when the work on this project really peaked, so we were kept so busy, we wanted to stay very focused. So we used... between us the code name rabbit so to to really to really indicate that there's a need to discuss the topic now right and one had to use the the code name to access us during uh this big time so i thought that was a funny anecdote to mention yeah um there was also uh still on the collaboration part the collaboration with the partners that you heard before so epaa in the European Commission who supported the project from the start and also have been working with us to promote it and ensure its success.
- Emmanuelle Charton
Yes, and the collaboration with the EU and EPAA culminated in an event which took place in February 2023 at Valentine. And it was a great opportunity because it took place in Brussels, so it was in-person meeting, it was also hybrid, but the EU Commission opened its doors to this event. So we were in the premises of the EU Commission and it was a joint event between EDQM and EPAA and there we announced about the publication of all these texts. It was a consultation phase at the time. And we invited also stakeholders from the whole world, from pharmacopoeias also outside Europe, to exchange about the European pharmacopoeia vision for the future. And it was also an opportunity to train stakeholders on how to use the MAT, because it's a test that you need to be mastering to be able to perform the test. It was both a matter of communicating on the project that Gwenaël describe, but also on making sure that this was understood outside Europe to promote the action outside Europe. And the outcome was very positive. In fact, we realized that the approach taken by the European Pharmacopeia was seen very positively. Of course, not all pharmacopeias would be ready to go because Maybe, for example, one of the reasons is they don't have the MAT described, so they do not have all the tools yet in their respective pharmacopoeias, but at least the approach was seen very positively from the outside. So from this event, we published a scientific article, and we consider this as a milestone, really.
- Presenter
All right. And based on the timeline that exists for phasing out, when... Can we expect the rabbit pyrogen test to fully stop being applied in Europe?
- Emmanuelle Charton
Yes, so we go on to also the climax of this whole exercise, which happened in June 2024.
- Gwenaël Cirefice
Yes, so in June, and that was the outcome of this exercise, the Open Pharmacopoeia Commission and the 39 delegations, so the delegation representing the 39 countries of the Open Pharmacopoeia, adopted about 60 texts that were revised to remove the reprogen test. and together with the new chapter on pyrogenicity. This really marks the end of the rapid pyrogen test. The new chapter and the revised text omitting the rapid pyrogen test are to come into force in July 2025.
- Emmanuelle Charton
Okay. Then to complete your question, this will be followed by the complete deletion from the chapter on the 1st of January, 2026. There's still some approval to be performed by the Commission in the meantime, but it is our vision and it's going to be the reality that as of 1st of January 2026, there will be no possibility to address pyrogenicity using rabbits as far as the European Pharmacopeia is concerned.
- Presenter
All right, so I assume this is considered a big achievement for the three R's that we mentioned before, replace, reduce, refine, right?
- Gwenaël Cirefice
Absolutely. And this decision to stop the testing of rabbits, we think is a significant milestone for animal welfare. We could say it's a landmark decision and it aligns with the Convention of the Council of Europe on Animal Welfare that I mentioned earlier. Right. And also it reflects the EDQM's strong commitment to reducing animal testing. whenever this is possible in the quality control of medicines.
- Emmanuelle Charton
And I would say it's not only an achievement as far as animal welfare is concerned, but also scientific advance. Because obviously in the 21st century, there are ways to perform tests without using animals. I mean, these animal tests were used in the 70s. And since then, the progress of science has shown that there is a possibility not to use animals. So it's also a testimony of EDQM's commitment to promote the adoption of methods that are state of the art for the pharmacopoeia. SL.
- Presenter
All right. So, Emmanuelle, Gwenaël, thank you so much for explaining this to me and to the audience. And thank you once again for coming to the podcast.
- Gwenaël Cirefice
EM. Thank you. GW. Thank you.
- Presenter
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