Research Responsible for High Levels of Animal Testing

In fields of research responsible for high levels of animal testing, hi-tech alternatives can replace, significantly reduce or minimise suffering. Not only more humane, alternative methods are quicker and cheaper. Yet still, millions of animals are used every year for quality control, toxicity testing and drug development purposes.

Quality control of vaccines

25% of animal testing in the UK is for quality control purposes! In 2005 a working party set up by APGAW called for more to be done to develop alternative methods for testing vaccines to replace the use of animals.^[i] In Europe, regulatory bodies require many types of vaccine to undergo testing to assess the potency and safety of each batch. This necessitates testing on large numbers of mice, guinea pigs, rabbits, and non-human primates.

However for some vaccines, including those for Pneumonia, Meningitis and HPV-VLP, non-animal methods are used. The safety and potency of these vaccines is determined using a battery of non-animal analytical methods. Researchers believe this new approach to vaccine testing can be applied more widely, reducing the number of animal tests:

• Diptheria vaccine

In 2002 researchers from the Netherlands Vaccine Institute outlined in Vaccine journal how the use of in-vitro and analytical methods could also be applied to vaccines that previously relied on animals. Research into this new “consistency approach” continued and in 2010, researchers concluded that non-animal methods could be used to test diphtheria vaccine.^[ii] These findings have been further substantiated by UK’s National Institute for Biological Standards and Control ^[iii]• Whooping Cough vaccine

In 2010, the UK’s National Institute for Biological Standards and Control, noted that in-vitro (non-animal) tests could be used to replace the in-vivo (animal) histamine sensitisation currently used for whooping cough vaccine^[iv].

• Tetanus toxoid vaccines

In 2002 the French Agency for Health Products (AFSSAPS) developed an alternative that could be used to replace quality control animal testing of the tetanus toxoid vaccine.^[v]

In April 2011, The European Vaccine Manufacturers (EVM) noted “In vitro tests (non-animal) are better analytical tools than in vivo tests (more powerful, reproducible, quantitative, appropriate for multiple antigens, inexpensive, and saving animals.”^[vi] It’s clear to see these new techniques hold enormous potential.

It’s now a matter of securing regulatory approval, before these methods can be implemented into mainstream practice, which is easier said than done!

The process of regulatory approval is a lengthy one. In a joint ECVAM/EPAA workshop held in 2010 participants noted some of the hurdles in implementing the consistency approach. Regulators in different parts of the world have different testing guidelines. This leads to different testing protocols in license applications for a given product depending on market location. The workshop concluded there was a need for close co-operation and the involvement of regulatory bodies^[vii].

Drug development: costly in terms of finance, time and animal welfare

Currently pre-clinical testing is required on two species, dogs, non-human primates, rabbits, rats and mice are amongst the species used. Tests to evaluate the safety and efficacy of pharmaceutical compounds can range from a few days to 3 months in duration, and increasingly, genetically modified animals are used. In 2010, 3396 beagles were used in pre-clinical drug development. A necessary evil, some might say? And yet typically 92% of compounds which pass animal tests then go onto fail in subsequent clinical trials!

If the drug does make it onto the market, the costs incurred in development are passed onto customers and the National Health Service. In times of economic restraint this surely has to be a consideration. The process of drug safety testing which has evolved over the years is unwieldy and wasteful, dependent on inordinately high levels of animal testing that is of limited value. It’s time for change....

UK companies developing alternatives to replace and reduce

Alternatives hold the potential to significantly reduce animal testing and deliver quicker, more human-relevant data. Each of the following UK companies are pioneering hi-tech techniques, proving there are better, cheaper, humane routes to drug development.

• Gentronix –“new, rapid, high-throughput and accurate assays”

Testing the genotoxicity of new drugs using animal models is costly in terms of time, typically taking 1-8 weeks, and money. “Regulatory tests cost about £36,000 per compound. That is an expensive way to find problems in late preclinical development.^[viii]” Not to mention the high toll in animal lives. In contrast non-animal genotoxicity screening methods generates results in 48 hours, using only a small amount of compound and at a fraction of the price, at about £500 per compound.

• Biopta - Human Tissue based tests

Using surplus tissue from surgical procedures it is possible to better predict the effects new drugs have on the human body. For instance it’s possible to tell whether a given compound will induce high or low blood pressure; or affect hormone levels using human tissue samples. Furthermore the data obtained is human relevant. As the graph ^[ix] clearly demonstrates, there can be vast differences in the results obtained through human and animal (canine) studies.

Human tissue testing has huge potential, human relevant and cost effective it’s hoped this technique will become a routine part of the drug development process. However before this can happen, measures must be put in place throughout the NHS to obtain consent for the use of surplus surgically removed tissues.

At the moment surplus tissue is made available from only 1-2% of surgical procedures.^[x] Up to 600,000 specimens are discarded each year, these tissues represent a valuable resource that is quite literally being thrown away.

• Xceleron - Microdosing

Microdosing involves giving minute doses of new drugs to human participants, to monitor effects. Over the past 10 years microdosing has proved to be a useful tool quickening drug development. Cost effective and fast, microdosing provides a good indication of the safety and efficacy of a new drug in humans, early on.^[xi] Enabling researchers to concentrate on new drugs most likely to meet clinical requirements and avoid costly and lengthy research into compounds that will fail in later clinical studies.

Toxicity testing of Botulinum Toxin

Every batch of botulinum toxin necessitates animal testing. In recent years, the increasing popularity of botox as a cosmetic preparation has seen significant increases in the number of LD50 tests.[i] In this crude test botulinum toxin is injected into the stomachs of mice, and over a 3-4 day period the mice suffer paralysis, breathing difficulties and ultimately suffocation. More...

Researchers have developed alternatives to the LD50 test, speaking in 2010 Dr Dorothea Sesardic, of the NIBSC noted…
“Alternative methods which reduce, refine and replace the use of live animals in testing botulinum toxin therapeutic products are now available. Validation and adoption of these methods will depend on the cooperation of regulatory agencies, manufacturers and validation centres, and will ensure that the 3Rs become an accepted and routine aspect of therapeutic botulinum toxin product testing.”

Reversing the trend: putting the 3Rs first

EU Directive 2010/63/EU adopted in September 2010, requires that researchers implement the 3Rs: replacement, reduction and refinement, as a matter of good practice. It’s now up to regulators, the scientific community and policy-makers to provide the support, funding and political will, to quicken the pace towards non-animal alternatives.

^[i] British Veterinary Association. (2005). Call for alternative methods for testing vaccines. Veterinary Record 2005;156: 394-395.
^[ii] Metz, B., Hendriksen, C., Jiskoot W., Kersten, G. (2002). Reduction of animal use in human vaccine quality control: opportunities and problems. Vaccine, 20: 2411-2430.
^[iii] Coombes L, Stickings P, Tierney R, Rigsby P, Sesardic D. (2009). Development and use of a novel in vitro assay for testing of diphtheria toxoid in combination vaccines. J Immunol Methods; 350:142e9.
^[iv] Yuen, CT., Horiuchi, Y., Asokanathan, C., Cook, S., Douglas-Bardsley, A., Ochiai, M., Corbel, M., Xing, D. (2010). An in vitro assay system as a potential replacement for the histamine sensitisation test for acellular pertussis based combination vaccines. Vaccine. May 2010, 7;28(21):3714-21.
^[v]Prieur, S., Broc, S., Gal, M., Poirier, B., Fuchs, F. (2002). Development of an in vitro potency test for tetanus vaccines: an immunoassay based on Hc fragment determination. In Brown, F., Hendriksen, C., Sesardic, D., Cussler, K. [editors]. Advancing science and elimination of laboratory animals for development and control of vaccines and hormones. Developments in Biologicals vol. 111, p.37-46.
^[vi] Ponsar, C., Chapsal, J., Scharton-Kersten, T. (2011). 3Rs and the consistency approach for human vaccine control: EVM Point of View. Workshop on the Application of the Three Rs and the Consistency Approach for Improved Vaccine Quality Control, Brussels, 7 April 2011, European Vaccine Manufacturers.
^[vii] De Mattia, F., Chapsal, J., Descamps, J., Halder, M., Jarrett, N., Kross, I., Mortiaux, F., Ponsar, C., Redhead, K., McKelvie, J., Hendriksen, C. (2010). The consistency approach for quality control of vaccines – A strategy to improve quality control and implement 3Rs. Biologicals. Volume 39, Issue 1, January 2011, pages 59-65.
^[viii] Gentronix, 2011. A Compelling Economic Case: Current genetic toxicity strategies waste money. [Online] Available at http://www.gentronix.co.uk/Genotoxicity/ACompellingEconomicCase/tabid/56/Default.aspx[accessed 19 April 2011]
^[ix] Moss, E. (2011). Blood vessel myography: a tale of two techniques. [Online] Available at http://www.biopta.com/blog/index.php [accessed 19 April 2011]
^[x] Bunton, D. (2011). The use of functional human tissues in drug development. Cell and Tissue Banking. volume 12, Number 1, p.31-32.
^[xi] Smith, D.A. (2011) The debate is over: AMS provides the route to better drug development paradigms / protocols. Bioanalysis, February 2011, Vol. 3, No. 4, Pages 391-392.