Vous êtes sur la page 1sur 24

ANIMAL

TOXICOLOGY STUDIES
(NON-CLINICAL STUDIES)

SWATI SARIN
WHY STUDY TOXICOLOGY???

Benefit –risk ratio can be calculated

Prediction of therapeutic index

Therapeutic index= Maximum tolerated dose


Minimum curative dose

Smaller ratio, better safety of the drug


WHY DO WE REQUIRE NON CLINICAL
STUDIES IN ANIMALS BEFORE
ADMINISTERED TO MAN??

 Pharmacological effects are same in man as in animals

 Toxic effect in species will predict adverse effects in man

 Giving high doses in animals improves predictability to man

 Risk assessment can be made by comparison of toxic doses in


test species with predicted therapeutic dose in man
PHASES OF DRUG DEVELOPMENT
MOLECULE
(ANIMAL MAN)
PRECLINICAL PHASE I PHASE II PHASE III PHASE IV
PHASE I

Patients Patients
Healthy subjects Patients
None Large scale Large scale
Safety and Small scale post-marketing
multicentre
tolerability efficacy studies studies studies

Genetic toxicity
Genetic toxicity
(in vivo)
(in vitro)
Repeat dose Chronic (long term) toxicity testing
Non-clinical

Single / repeat dose


toxicity testing +
toxicity studies
+ Bioanalysis / Toxicokinetics
+
Bioanalysis /
Bioanalysis /
Toxicokinetics Reproductive Toxicity Testing As required
Toxicokinetics
(fertility and pre/post natal)
Drug Metabolism
Safety Pharmacology
Carcinogenicity studies
Reproductive
Drug Metabolism
Toxicity Testing Drug Metabolism
(teratogenicity)

Lead candidate Entry to man Patient studies Product Approval


Identified (IND / CTA) (NDA/MAA)
GENERAL PRINCIPLES IN TOXICITY
STUDIES

Studies should comply with GLP


Performed by trained and qualified staff

Use of standardized and calibrated equipment

SOP’s followed in laboratory tasks

All documents should be preserved for minimum 5 years after


marketing of the drug
TOXICOKINETIC STUDIES

Generation of Pharmacokinetic data to access systemic


exposure achieved in animals

Relation to dose level and the time course of toxicity study

To support choice of species & Treatment regimen

Design on clinical studies accordingly


RELEVANT TEST MODELS
 Pharmacodynamic responses

 Pharmacokinetic profile

 Species, sex, age of experimental animals

 Susceptibility, sensitivity and reproducibility of test


system

 In vitro: Isolated organs, tissues cell-cultures

 Mechanism of effect in vivo


TYPES OF TOXICOLOGY STUDIES
Systemic toxicology studies

Single dose studies Repeated dose studies

Reproductive toxicology studies

Male fertility Female reproduction & Developmental


studies
Local toxicity studies

Hypersensitivity studies

Genotoxicity studies

Carcinogenicity studies
A. SYSTEMIC TOXICOLOGY STUDIES

a) SINGLE DOSE STUDIES/ ACUTE TOXICITY

Preliminary Definitive

• Maximum Non • MTD and MLD


Lethal dose determined
(MNLD) determined • Evaluate effects
• Target organ of toxicity
may be determined
PRELIMINARY STUDIES
METHOD

 Single dose tested in 2 rodent species


 2 routes of administration
 Oral dosing of 2g/kg or 10 times of normal human dose
 Observation for 14 days after dosing
 MNLD established
 Symptoms , signs reported
 Microscopic and Macroscopic evaluation
DEFINITIVE STUDIES

METHOD

 Group of 20 animals of either sex dosed at MNLD


 5 animals of each sex are observed for 48 hr and conduct
autopsy for early pathological changes
 Remaining 5 of each sex are observed for 14 days
 MTD and MLD established
 Signs of intoxication or recovery, changes in body weight,
pathological changes
 Complete macroscopic and microscopic examination
 Target organs can be identified
b) REPEATED DOSE STUDIES/SUB-ACUTE
OR CHRONIC TOXICITY
 Two mammalian species(one should be non-rodent)
 Long duration studies (30-180 days)
 Dose is dependent on dose-escalating studies
 Drug administered by clinical route
 Parameters monitored and recorded are:
 Behavioral

 Physiological

 Biochemical

 Microscopic observations
B. REPRODUCTIVE TOXICOLOGY STUDIES
a) MALE FERTILITY
METHOD
One rodent species(rat)

3 dose groups taken


(each with 6 adult males),
1 control

Drug treatment by clinical route for 28-72 days


Mated with females in 1:2 ratio

Females getting pregnant should be examined


After 13 days of gestation

All male animals sacrificed

•Weights of testis, epididymus recorded & examined for


their histology
•Sperms examined for motility & morphology
b) FEMALE FETILITY

Drug administered to both males (28days) and females (14


days) before mating
Segment I Fertility and general reproductive
performance study
Implantation
Segment II Teratogenicity Embryogenesis

Segment III Peri and post-natal study

Fertility and early embryonic


development (rat)
Embryo- foetal development
(rat & rabbit)
Post natal development (rat) (post natal
survival of offspring), growth parameters,
vital senses, behavioral effects

15
C. LOCAL TOXICITY STUDIES

 Required when drug is administered by special route


(other than oral) in humans

 Study design:
 2 species along with control used
 Dose dependent on dose escalating studies
 3 dose levels
TYPES OF LOCAL TOXICITY STUDIES
Dermal toxicity studies Rats & Rabbit
Local signs (erythema, oedema),
histological examination

Dermal photo-toxicity Guinea pig


studies Used in treatment of leucoderma
Examination of erythema &
oedema formation

Vaginal toxicity studies Rabbit or Dog


Observation of swelling,
histopathology of vaginal wall

Rectal tolerance studies Rabbit or Dog


Signs of pain, blood or mucous,
histology examination of rectal
mucosa
Albino Rabbit
Ocular toxicity studies Changes in cornea ,Iris &
aqueous humor, histological
examination of eye

Parenteral drugs For intravenous/


intramuscular/ subcutaneous/
intra-dermal injection
Sites of injection examined
grossly and microscopically

Inhalation toxicity studies


One rodent and non rodent
species
Acute , sub-acute and chronic
studies performed
Observation of respiratory rate
Histological examination of
respiratory passages, lung tissue
D. ALLERGENICITY/HYPERSENSITIVITY
TOXICOLOGY STUDIES
Guinea Pig Maximization Determination of Maximum non
test irritant or minimum irritant dose
Evaluation of Erythema and
oedema

Local lymph node assay Mice of one sex(either male or


female)
Drug treatment given on ear skin
Auricular lymph node dissection
after 5 days
Increase in 3h-thymidine used for
evaluation
E. GENOTOXICITY STUDIES
To detect early tumorigenic effects in cases of chronic illness

In vitro tests:


Test for gene mutation in Bacteria
Cytogenetic evaluation of chromosomal damage in
mammalian cells
E.g.; Ames’s Salmonella Assay detects increased number
of aberrations in metaphase chromosomes
DNA strand breaks, DNA repair or recombination,
Measurements of DNA adducts

In vivo tests:


Chromosome damage in rodent hematopoietic cells
E.g.; Micronucleus Assay
F. CARCINOGENICITY/ ONCOGENICITY
STUDIES

 Life-time Bioassays

 Carcinogenicity studies are performed on:

 Drugused for >6 months or frequent intermittent use for


chronic diseases

 Chemical structure of drug indicates carcinogenic potential

 Therapeuticclass of drugs which have produced positive


carcinogenicity
CONDUCT OF STUDY

Group sizes of 50 animals/sex at each of 3 dose


levels
Control group is of double size
Record for onset of tumor development
Usually carried out for 24 months in rats and 18
months in mice (life span studies)
EVALUATION OF RESULT

Incidence of cancers in control and test


 Trend towards increasing incidence with
increasing doses
Number of animals with single/multiple tumors
Macroscopic changes observed by autopsy
 Histopathology of organs and tissues
THANK YOU

Vous aimerez peut-être aussi