minipigs in the safety assessment of...
TRANSCRIPT
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Minipigs in the safetyasessement of drugs
Peter Glerup, DVM, MScChief Veterinary Surgeon
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Outline• Regulatory requirements• Choice of Non-rodent
species• Pigs and minipigs• The Göttingen minipig• Similarities to humans• Dose routes, study types
andexperimental procedures
• Conclusion
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Regulatory requirements
• A rodent and a non-rodent species should be used in safety asessment of new pharmaceutical products
• Dogs or non-human primates traditionally used• Focus on the minipig significantly increased
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Choice of non-rodent species
• Pharmacological responsiveness similar to man• Metabolic similarity• Physical or anatomical similarity• Comparative reasons (species previously used
for same class of compound)
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Pigs and minipigs
• 13 species of pigsworldwide(suidae)
• At least 11strains ofminipigs exist
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Why use minipigs?
• Size• Similarity to man• Growth in background data• Regulatory acceptability• Availability of high quality animals• Concerns about primates (availability, handling,
health status, zoonosis, emotive, costs)• Concerns about use of dogs (emotive, vomiting)
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Types of minipigs
• ”Natural”, ex. The Yucatan minipig• Purposely bred, ex. The Göttingen minipig
• Weight ranges35-55 kg: Yucatan micropig, Göttingen minipigs50-60 kg: Sinclair, Hanford70-90 kg: Yucatan minipig, Hormel minipig
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The Göttingen minipig I
• Bred in Germany (Göttingen) from the 1960´s• Purposely bred entirely for use in biomedical research• Breeding ”goals”:
- small size – easier to handle, less test item - ”non-pigmented” – use for dermal research
• Crossbred- Minnesota minipig- Vietnamese pot belly pig- German landrace
• Well defined health status
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The Göttingen minipig II
• Age up to 20 years?• Body weight at birth: 0.45 kg
Body weight at weaning: 2-3 kg• Sexual maturity
- males: 3-4 months (7-9 kg)- females: 4-5 months (9-11 kg)
• Body weight at age of 1 year: 25 kg• Fully grown body weight: 35 kg
- older boars up to 50 kg- older sows after multiple litter up to 60 kg
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Similarity to humans
• Structure of the skin• Gastrointestinal tracts• Cardiovascular system• Metabolism• Teratogenic sensitivity• Urogenital system (size, number of lobes)• Nasal cavity (two sides separated)
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Göttingen minipig skin
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Rat
Rabbit
Dog
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Gastrointestinal tract• Pigs and humans are omnivorous• Pigs have similar gastric cell types, villi and
secretions to humans• pH changes and transit
time in the small intestinesimilar to humans
• Colon arranged in series of coils
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Cardiovascular system
• Heart and great vessels similar to man (except left azygos vein)
• Coronary arterial blood supply to heart almost identical to humans
• Induction of myocardial infarction• Hypercholesterolaemia
and artherosclerosis
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Metabolism
• Minipig CYP 1A, CYP 2A and CYP 3A metabolise same test substrates as in humans
• 75% identical cDNA sequences• The minipig is a good model if these enzymes
are involved in the metabolism of the test compound
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Housing of minipigs
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Routes of dosing/study types
• Oral (gavage/capsule/tablet)
• Dietary• Inhalation/intranasal• Dermal• Injection (sc, im, iv, ip)• Continous iv infusion• Intravesical• intravaginal
• Single dose studies• Repeat dose studies• Teratology studies• ADME studies• Telemetri• Disease models
- diabetes- wound healing- parkinsonism- transgenic minipigs
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Dermal dosing I
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Dermal dosing II
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Local tolerance studies
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Oral gavage
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Subcutaneous dosing
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Intramuscular dosing
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Blood sampling
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Ophthalmoscopy
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Urine collection
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Electrocardiography
Normal sinus rhythm
Slight sinus arrhythmia
Occasionally wandering pace maker
Heart rate 90-164
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Anaesthesia and surgery
• Rapid gastric emptying- short fasting period
• Injectable anaestheticsfor surgery of short duration
• Inhalation anaesthetics recommended for longer procedures
• Endotracheal intubation requires special laryngoscope
• Anatomical differences (intestinal system, azygos veins)
• Post-surgical analgetic treatment(opoids, NSAIDs)
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Intravenous dosing techniques
• Sparse peripheral veins• Ear veins for single dosing• Surgical intravenous access recommended for
repeated i.v. dosing- Vascular Access Ports
• Infusion pumps connected to i.v. catheters used for continuous infusion
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VAP implantation
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Wound healing techniques• Skin and woundhealing of minipigs similar to humans• Adult animals can be used• Two models generally used
- split-thickness wounds- full-thickness wounds
• Studies individually designed (limited regulatory guidance)• Models of acute wound healing, not chronic
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Juvenile studies
• Paediatric development in focus• Age dependent pharmacokinetic, pharmacodynamic or
toxicological differences may exist• Development of main organ systems have important
structural and functional differences with age• Many practical aspects to be considered for each
individual species
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What is a juvenile pig?
• Humans- neonate from birth to 1
month- infant 1 month to 2
years- child 2-12 years- adolescent 12-16 years
• Minipigs- neonate from birth to 4
weeks- infant 4-8 weeks- child 8 weeks to
3 months- adolescent 3-5 months
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Juvenile studies, study design
• Time mated sows• Considerations for group establishment
- one litter = one group (genetic bias)- all groups represented in one litter (contamination risk)- cross fostering (time mating, induced farrowing)
• Study procedures starting from Day 7
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Teratology studies
• Minipig useful alternative to rabbits• First choice when metabolism is similar to man• Susceptible to human teratogens
(thalidomide, hydroxyurea, ethanol and others)• Large litter size as compared to primates• Gestational period shorter than primates• Regulatory acceptance
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Uterus with fetuses
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Alizarin stained fetus
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Telemetri
• Cardiovascular safety pharmacology• Animal handling at dosing only• Transmitter placed s.c. in flank• Blood pressure
transducer placed in a. femoralis
• Wires for ECG placed s.c. over thorax
• Battery life time of transmitter
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Conclusion
• The minipig shows many similarities to man• The minipig constitutes a suitable model
for a wide range of non-clinical studies• All general routes of dosing can be used• Anaesthesia of minipigs is a non-complicated procedure• Many surgical procedures are practically feasible• The minipig is accepted by regulatory authorities
worldwide as a useful non-rodent species
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The end...