making sense animals of animal research

7/29/2019 Making Sense animals of Animal Research

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Making sense ofanimal research

REPLACE

REDUCE

REFINE


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Development Phase Questions Addressed Duration NuCompo

Discovery o New Drug What target enzyme or protein? What test system to show drug-proteininteraction? What are chemical lead structures?

3 years

Optimization What is structure-activity relationship? What molecules have not onlyintrinsic activity but also drug-like properties (absorbability, metabolicstability)?

2-3 years

Preclinical Development How can kg-amounts eciently be synthesized? What are suitableormulations? What is pharmacokinetic metabolic and saety proleo the drug candidate?

1-2 years

Clinical Testing What is tolerable dose range? What is dose-eect relationship?Is drug suciently sae at ecacious doses?

4-5 years

Registration What are sources o variability? What is adverse eect prole? 1 year

10

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Drug Development and the 3Rs


3/16Making sense of animal research

The pace o medical research is ast, but there are still many heachallenges ahead. Animals continue to play an invaluable role

meeting these challenges - both in experimental research, and

ensuring maximum saety o treatments beore their use in

mans.

The process or developing a medicine does not start with anim

research, which represents only about 10% o the whole R&D pr

ess. It is only undertaken when it is deemed necessary to addres

specic scientic question which cannot be answered ound thro

other routes.

Most o the research and development eort goes into non-anim

techniques and human clinical trials. But where animals are

volved, it is because their role is crucial and indispensable - a

will continue to be so or continued improvements in health care

Progress in science is constantly delivering new and better the

pies - or humans and or animals. And the same progress is a

providing more and more alternatives to the use o animals.

This publication gives a glimpse o the process o creating n

medicines. It also spells out how medicines researchers are at

same time discovering methods to rene, reduce, and in some c

es even replace the use o animals.

It discusses what can be done now without having to use animal

research, what cannot yet be done, and what the prospects are

the uture. Above all, it illustrates the close link between prod

ing new medicines and discovering new ways o developing me

cines. Driving scientic advances orward makes sense or anim

research - and paves the way or alternatives.

intro

Making sense of animal research

Tools and 3Rs Principles Applied

Non animal research:

Computer modelingHigh throughput screeningmaging techniques

n vitro studies:

Cell culturesTissuesOrgans

Animal and in vitro tests:Ethical review; Improved housing and care;Rened sampling techniques; Smarter study design; Imagingechnologies; Chip technology; Microdosing; Stem cells;nternational harmonization; Replacing higher by lower species; etc

Human clinical trials:

Phase I, II and III (saety, ecacy, variability)


4/164 Making sense of animal research

in the past

Many o the medical advances o the past are taken or granted to-

day. But they would not have happened without animal research.

For well over a century, research on rats, dogs and pigs has helped

to nd new treatments or conditions as diverse as heart disease,

inections, brain disorders and arthritis.

Poliomyelitis has been wrecking human lives or ages, and the last

century saw epidemics that crippled and killed hundreds o thou-

sands around the world. Now the disease is close to being eradi-

cated worldwide. New medicines have made this transormation

possible - vaccines that have been developed over some orty years

o research using monkeys and mice.

Breast cancer is the most common cancer among women. Over

the last twenty years, successul new treatments have started to

emerge. The radically improved survival rates that these medicines

oer are the direct result o extensive research including animal

studies.

Asthma is the most requent serious childhood illness in the devel-

oped world, where it still causes thousands o deaths every year.

But the inhalers commonly seen in schools today deliver medicines

that control the condition and allow suerers to lead a normal lie.

No animal research, no medicines

These medicines are available only because their ecacy and sa

ty have been assured by studies involving animals.

For centuries, the lie o diabetics was short and miserable. T

disease was inescapably lethal, and death could be deerred o

by a strict diet, until the lie o a 13-year old in Toronto was sa

in 1922 by an injection o a puried extract rom isolated pig p

creases. Evaluation o this innovative approach on a emale bea

led to the understanding o insulins eect to lower blood sugar

nding that has allowed millions o patients since then to surv

and lead normal lives.

The success story o transplantation started in 1905 with the

successul human transplant, a cornea, in 1905. A breakthrou

was the rst transplantation o a human heart by Christian Barn

in 1967. Since then it has become common to replace heart valv

kidneys, hearts and bone marrow. Every transplant surgeon nee

to study the complicated techniques in animals prior to condu

ing surgery on human patients. Also experimental animals w

essential to understand the immunological process o transpl

rejection and to develop drugs to prevent rejection. Such medici

are still being developed and investigated in rodents, dogs a

monkeys.

4 Making sense of animal research



and in the present

Many questions can still be answered only by research that makes

use o animals. Despite the advances in computer modelling and

in alternative methods such as the use o single, isolated cells, the

development o diseases or physiological processes cannot always

be suciently studied with these techniques. Animal studies oer

hope to millions o people with conditions ranging rom stroke to

spinal cord damage, and rom sickle cell disease to malaria. The

use o monkeys is opening up new possibilities or deep brain

stimulation or Parkinsons Disease; mice are making a critical

contribution to developing new treatments or adult leukaemia and

lymphoma; rabbits and cattle are both playing a part in producing

cervical cancer vaccines, and goats in developing blood clotting

agents rom milk, while bird fu vaccine development is dependent

on the use o chickens and errets.

Developing treatments or Alzheimers Disease requires an un-

derstanding o the behavioural changes brought on by the disease.

Computer programmes and studies o individual cells are not

enough or this. Scientists also make use o animal models, to gain

new insights into the causes and to develop new treatments. Mice

were used to prove that when injected directly into the brain, anti-

bodies against a plaque protein retarded its growth. Gene therapies

that have qualied or clinical trials in humans have rst been stud-

ied in animals,.

Cystic Fibrosis is the most requently occurring lie threatening

genetic disease in Europe. Sticky mucus blocks the lungs and di-

gestive system. One out o 30 Europeans carries the gene caus-

ing the disease and 30,000 children and young adults live with it.

Some patients live until theyre 40 years old, others only 4 years,

depending on where they live in Europe. No cure yet exists, but the

use o transgenic mouse models is expanding our knowledge o the

disease every day. This will be useul or testing lung treatments.

Several ways o correcting the fuid transport have already been de-

veloped in mice.

Mice are used or initial evaluation o potential HIV vaccines. Butsince the virus does not inect mouse cells, promising vaccines are

tested urther on macaque monkeys, which have immune systems

more similar to man, allowing an accurate assessment o cellular

responses.

Epilepsy is a neurological disorder characterised by seizures t

result rom excessive electrical activity in the brain. Epilepsy-pr

rats have been used to identiy a protein that triggers seizures. T

researchers used a technique that measures brain activity. A

a chemical called ethosuximide that protects young animals r

seizure has been identied.

Work with mice and dogs is deepening our understanding o m

cular dystrophy, which still causes muscle wasting and early de

in humans. In particular a naturally- occurring condition in so

dogs closely mirrors the human disease mechanism, providing

invaluable model or clariying how the disease disrupts norm

body unctions, and or developing potential therapies.

Making sense of animal research



As part o their search or new medicines, all researchers are alsolooking or better ways o working. Progress in scientic knowledge

and in technology continually oers new opportunities or improv-

ing methods and results.

Revolutionary advances over the last 20 years include gene se-

quencing, the increasing precision o modern MRI scanners, high-

speed computer modelling and screening and better understanding

o cell behaviour. These techniques have in many cases reduced the

need to use animals, or, where animals are still needed, have re-

ned the way that studies are conducted, oten both reducing stress

and delivering better-quality data.

Research currently underway is exploring the use o stem cells

cultivate human liver and heart muscle cells, allowing replacem

and reduction o some animal studies.

Implants or the treatment o bone ractures used to be studied

sheep. Since it became possible to culture cells to create new t

sue, much o this work can now be done without using animals. C

tures o bone cores provide tissue that acts like live bone, which

be used or testing strength, and also allows more ecient stud

processes in bone tissue.

To develop repellents against ticks, cats were routinely used as hanimals so that ticks could eed on their blood. Now scientists h

developed articial eeding membranes that simulate the skin

host animals. In addition, this method makes it possible to ma

more precise measurements o the eect o dierent substan

added to the blood in these articial systems.

Non-invasive imaging techniques are used or research into dise

and or in vivo saety assessment o new compounds. They h

helped to reduce the number o animals by up to 80%. In addit

these techniques can enhance the quality o data and statisti

power o a study. Imaging techniques allow continuous observat

o a single anaesthetised animal, and thereore do not require k

ing a number o animals at dierent stages o the study or autoinvestigations. They provide inormation about structure and u

tion o organs, the progress o a disease and how the same te

nique might unction in humans.

Progress in science leads to progress

in alternative testing



Studies o the human genome, breakthroughs in high-speed inor-mation processing, or the promise o nanotechnology all hold out

the prospect o new or improved medicines.

Possibilities o success in meeting health challenges such as

emerging diseases, or in new avenues o research to investigate,

are revealed only as our knowledge grows.

Against this rapidly evolving context, animals continue to play an

invaluable role - both in experimental research, where they allow

investigation o a medicines potential, and in saety evaluation, to

reveal any harmul eects beore use in humans.

The process or developing a medicine does not start with animal

research. It is only undertaken when it is deemed necessary to an-

swer a specic scientic question, whose answer cannot be ound

through other routes.

Science is still moving ahead

and researchers are still seeking

alternatives.

As part o the eort to discover new medicines, researchers are

constantly seeking ways to improve their research methodology

to improve eciency and results. This means that renement, re-

placement and reduction in animal research are part o the scien-

tic process.

It is not in researchers interest to use animals unless they are ab-

solutely necessary. Using animals in experimentation is very cost-

ly - not just because o the breeding and acquisition costs, or the

complexity o procedures, but also because o the high standardso husbandry and welare that animals require and deserve. In ad-

dition, the scientic data that have in the past come mainly rom

animal research can increasingly be derived rom new techniques

that oten provide a higher degree o precision in results, because

they are not infuenced by the inherent inter-animal variability or by

environmental actors. And the use o animals also raises continual

ethical questions, requiring the most rigorous justication in per-

sonal and proessional terms.

So major investments are also made in the development o alter

tive methods - to reduce the number o animals used, to rene

way they are used so as to minimise discomort or distress, a

where possible, to replace animals. This widely supported strat

o reduction, renement and replacement is requently reerred

as the 3Rs approach.

1.000.000

screening

isolated

target

10.000

in vitro

cellular

systems

100

in vitro

in vitro

ADME

5

in vivo

animal

testing



Laboratory mice and rats spend their lives in a cage. Enriching their environment, withshelters, shredded paper, pieces o wood, or climbing opportunities, allows them to ex-

plore, to build nests, and to nd reuge - to exhibit normal activities and thus to reduce

stress. Enrichment has been shown to reduce inter-animal variability in response to

experimental conditions,

and in some cases may

reduce the number o ani-

mals required to provide

inormation.

Work in progress on alternatives

Thsans initiatives are nerway right arss Ere t eve aternatives t existingresearh meths. Sme aternatives he t ree the nmber animas se, an even

t reae them entirey in sme reres. A these erts are aime at rening the

researh ress, an, where the se animas remains neessary, at maximising mrt

an minimising istress. Reent exames aternative meths evee by the harma-

etia instry ine the se in vitr iver es t assess metabism an eiminatin,

syntheti anima rtein t artiay reae anima tisses, an b es r yrgeniity

testing. They as ine the se mter tehngies in the eary isvery ress,

measring ner-hemias in the mammaian brain sing in viv mir iaysis, an rea-

ing ginea igs with a e-base assay t assess the ratin atin rgs r the treat-

ment asthma.

REFINEMENT

3 reducing stress in animals


9/16Making sense of animal researchMaking sense of animal research

Companies operate their own internal ethical review systems that oten exceed any legalrequirements; they establish high standards or housing o experimental animals; they

set up supervision mechanisms or all animal research; and they und research into via-

ble alternatives, and promote awareness o alternatives through awards and education.

REFINEMENT, REducTIoN ANd REplAcEMENT

3 ensuring ethical procedures

While in vivo models allow reliable measurement o alterations in articular

cartilage, the cellular mechanisms controlling those changes are dicult

to examine. In-vitro models oer a more direct way to study the cellular re-

sponse to articular loading under controlled circumstances. Because such

in-vitro studies are a closed system, the experimental conditions can be

controlled more precisely, and inter-individual variability can be reduced.

REducTIoN ANd REplAcEMENT3 greater accuracy from in vitro

systems



The design o studies involv-

ing animals is constantly being

improved, or instance by the

prior denition o a humane

endpoint - the point when the

test can be stopped because

its objectives have been met and beore the animal suers signicant distress. This

presupposes adequate methods or measuring distress, and the attention o ully

trained personnel. Careul consideration o study design can also reduce the number

o animals required or a trial. The use o statistical support helps to ensure that the

number o animals is as small as possible, while providing scientically meaningul

results.

Small implants allow remote monitoring o heart rate and blood pressure in animals

used in research throughout an experiment. This means animals dont need the

more stressul (and laborious) removal rom their social environment or repeated

scanning, and boosts the quantity and quality o data, thus reducing the number o

animals required to produce meaningul results.

REFINEMENT ANd REducTIoN

3 information from implants

REFINEMENT ANd REducTIoN3 smarter study design

10 Making sense of animal research



REFINEMENT ANd REducTIoN

3 gentle pain measurement

To investigate the capacity o new medicines to relieve pain and improve mobility in

arthritis, companies use a new system o imaging and analysing how an aected

animal walks, so as to immediately detect any subtle changes ollowing treatment,

beore the changes progress to a more painul condition. This is less stressul or

the animal than older methods, and can provide more accurate data using a smaller

number o animals.

To protect the unborn child, new medicines need to be studied to

make sure they would not damage the embryo i used by a pregnant

woman. Current international guidelines require studies on pregnant

animals, but some companies are employing an in-vitro embryonic

stem cell test. This cannot replace the ormal test on live animals,

but it can reduce the number o such tests, by helping exclude com-

pounds that are not suited or urther development and thus avoiding

unnecessary testing.

REducTIoN3 early in vitro safety testing



The aiatin 3Rs rgrammes is an integra art the harmaeti

instrys ay t ay researh ativity. Every many invve in rg evment is keen t ree reiane n anima ata, an t maximise the va

ata by imrving weare an by stanarising qaity reres wh

animas have t be se. Many manies reqire eah their researh si

t reate an anna an enhanement, vering imrve hsing n

tins, ess stress r invasive researh tehniqes, r better sty esign

ree anima se.

Unnecessary duplication o animal studies is being reduced by collaboration be-

tween European pharmaceutical companies and animal welare charities. They are

centralising existing but widely dispersed data on the characteristics o the excipi-

ents used as vehicles or test substances, and making these data available to scien-

tists who are planning animal studies. This will signicantly reduce the need to study

known excipients in animals.

REducTIoN

3 sharing data between researchers

Primary cell cultures are increasingly used to rank or select compounds in the early

development stage, making it possible to discard unsuitable compounds beore any

animal research is undertaken. In vitro human hepatocyte cul-

tures are used to assess how a compound will be metabolised

and eliminated rom the body, or instance, and new automated

tests also improve accuracy. As a result, compounds are more

precisely selected at the early stage o research, reducing theneed or animal studies.

REducTIoN3 early elimination of unsuitable compounds



The use o cell cultures instead o live animals oers huge scope as

an alternative test method. But or scientic validity, some studies

have to be conducted in a complete living organism, since cells do

not experience anxiety or endure diarrhoea or suer rom neuro-

behavioural disorders.

Any reduction in the number o animals used in a particular proce-

dure also has to be made in a way that does not reduce the scien-

tic validity o the test. I too ew animals are used, the results may

be invalid, and the test will have to be conducted again - result-

ing in the use o more rather than less use o animals. Statistical

input can help to optimise study design to deliver the inormation

required using the minimum number o animals.

Even where valuable alternatives exist in principle, technical limita-

tions may sometimes prevent their implementation in practice. For

instance, scanning oers huge potential or renement, reduction

and even replacement o some animal testing procedures. Until re-cently scanners were developed principally or use with humans, and

were thereore o a size and design unsuitable or rodents. Techno-

logical advancements in recent years helped to miniaturise these de-

vices or use on mice, with excellent results.

Regulations can also be a barrier to the use o alternatives. Animal

study requirements are clearly described and controlled in Europe -

and so is the use o alternatives. Once an alternative has been devel-

oped and validated, it still has to be ormally accepted in the context

o the international registra-

tion o a medicine.

It can take 5-10 years to vali-date and implement a new

method or legally required

studies (and cost more than

500,000), and 3-5 years to

validate and implement a new

method or basic research.

The outcome o these long and

expensive processes is not al-

ways successul.

Legislation is not static. The relevance o regulatory requireme

is also reviewed. For instance, the LD50 test (a study to nd

dose o a compound that causes death in 50% o animals treated

now no longer required. More recently, pharmaceutical compan

have demonstrated that single-dose acute toxicity studies on a

mals are no longer necessary beore rst administration o a n

medicine to man, because the data they provide can be obtai

rom other sources. As a result they have already reduced the us

this method in their internal procedures. However, this requirem

still has to be ormally removed rom regulations beore it cease

be a mandatory elements in the authorisation procedures or n

medicines.

European and national pharmaceutical laws set specic regulat

obligations or saety, quality and ecacy that can currently only

ullled with the results o animal studies. Saety and regulatory

quirements are continuously being reviewed, and this unortuna

has requently resulted in an increase in requirements.

Europes legislation protecting laboratory animals sets very h

standards. Additional requirements can be justied only when t

bring tangible improvement in animal welare and where they

not compromise patients access to essential treatments. Und

tightening o regulation in Europe could make things worse. For

stance, banning the use o non-human primates in Europe wo

not stop this type o research, but would merely drive it to m

distant territories where Europe has no mandate to infuence

standards o animal care and use.

Over-regulation might severely reduce the research and devel

ment o new medicines in Europe. Indeed, although it representsmall part o the ten years or more normally required to develo

new medicine, the use o animals in pre-clinical studies is pivo

Driving animal research out o Europe would also drive out ot

high value added R&D processes - and reduce the EUs scien

base, exporting research jobs, and leading to research conduc

outside the EUs regulatory scrutiny.

Complex science

and complexprocesses

In meiine, as in a sienes, nthing is sime - an neither isthe evement aternatives t the se animas in meiines

researh. The imitatins are sienti, tehnia, regatry, an

gistia.



Where next for alternatives?

Meanwhile, pressures or ever-saer treatments tend to streng

en and broaden saety assessment requirements. Testing o ea

batch o vaccines to be released on the market is still mandatory

public health reasons. In many cases this has to be carried ou

live animals. Until the law is changed, thereore, animals will c

tinue to be necessary.

And because the pursuit o research is an open-ended exercise

is possible that new developments will urther boost demand

the use o animals in testing procedures. Dramatic advances

biotechnology have made it possible to design genetically modi

animals, which can be purpose-designed or specic test pro

dures. Transgenic animals are helpul and sought-ater mod

particularly in basic research and especially since the mapping

the human genome. Researchers have now identied all the hum

genes, they still do not know the unctions o many o them. G

netically altered animals can help nd the answers, because m

human genes are ound in a similar orm in mice. The same te

nologies are creating the prospect o radical new treatment optithrough cell and gene therapy.

The availability o these techniques creates opportunities or n

research - to overcome continuing challenges such as cancer, A

and cardiovascular disease. This simultaneously drives the need

using animals, in basic and applied research, and in saety testin

The prospects are good or alternatives to play an increasing rolein the research and development o new medicines. Scientic and

technological advances are generating an ever-wider selection o

methods to rene, reduce and replace the use o animals. Mecha-

nisms are also in place to speed regulatory acceptance o alterna-

tives in saety and ecacy evaluation, and to standardise test re-

quirements more widely.

But there is currently no prospect o an end to the use o animals in

medicines development. Mice and rats are contributing to develop-

ment o stem cells or spinal cord and heart repair; work with mice

holds out the prospect o oral or inhaled insulin or type 1 diabetes,

o angiogenesis inhibitors or cancer and blindness, o gene therapy

or muscular dystrophy; and mice and monkeys (e.g. macaques) are

being employed in the development o malaria vaccine.

In addition to current unmet needs in the treatment o disease, new

diseases continue to emerge.


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The inescapable dilemma

So scientists, regulators, industry, and society at large ace an

ethical dilemma: is the use o animals justied in the interests o

a greater benet? At present, the societal consensus is that it is

justied where it is indispensable and where possible harm is mini-

mized.

But each new development in science brings with it new questions

o an ethical nature. As eective alternatives are developed and

validated, they should be employed. However, beore then can e-ectively be used, they need to be accepted by the regulators.

Industry and researchers too are constantly reassessing the bal-

ance between the development o new medicines and the develop-

ment o alternatives to animal research.

Advances in science lead to new opportunities, new challeng

new medicines, and to new alternatives - and some o these n

avenues can mean using ewer animals, while some may invo

the use o animals in new ways. Everyone involved - and that a

includes society at large - must play a part in refecting on how t

inescapable ethical dilemma is resolved.

I researhers isense with the se animas in eve-

ing new meiines, they w. N sientist wishes t infit any

istress r ismrt n any iving reatre. Bt at resent new

meiines een n anima researh. It is nethia an iega

t test r se a new sbstane in hmans witht rst ensring

maxima ssibe saety.


16/16

le paza BiingRe Trne 108 - bx 1B- 1050 BrssesTe: +32 (0)2 626.25.71e-mai: [email protected]

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making sense animals of animal research

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