GLP Toxicology Services

Back to Preclinical Services

PRE-IND STUDIES REQUIRED FOR PHASE I TRIALS

ACUTE TOXICITY

SUBCHRONIC TOXICITY

GENETIC TOXICITY

  • Ames reverse-mutation assay
  • CHO-HGPRT mammalian cell forward gene mutation assay
  • Cytogenetics assay in primary human lymphocytes
  • Micronucleus study in rodents

SAFETY PHARMACOLGY

  • There are ~ 15 short term studies in this series
  • Assess adverse effects of compounds on the cardiovascular, respiratory, central and peripheral nervous system, renal and gastrointestinal functions.

Studies normally recommended are dog cardiovascular, rat pulmonary function, Irwin CNS behavior study in rats, HERG Tail Current Test.

TOXICOKINETICS

Can be included in the Subacute studies

 

POST-IND STUDIES REQUIRED FOR AN NDA

CHRONIC TOXICITY

  • Daily dosing of rodents 6 months, non-rodents 12 months
  • Daily dosing via the clinical route
  • Two species (rodent and non-rodent)
  • Three dose levels plus controls
  • Clinical, toxicokinetic, clinicopathologic, and morphological evaluations

CARCINOGENICITY

  • Daily dosing for two years
  • Clinical route
  • Two species (rat and mouse)
  • Three dose levels plus two controls
  • 50-60 animals/sex/group
  • Assess carcinogenic potential of material
  • May substitute a six month transgenic mouse study for the 24 month mouse

REPRODUCTIVE TOXICITY

Fertility (Male and/or Female) - Segment I

  • Fertility and early embryonic development to implantation
  • Rat dosed via the clinical route
  • Three dose levels plus controls
  • Dose males 4 weeks prior to mating, females 2 weeks
  • Assess maternal toxicity, development and maturation of gametes, estrous cycles, mating, fertility, and effect on implantation

Embryo-fetal Development - Segment II

  • Embryo-fetal development
  • Two species, rat and rabbit does via the clinical route
  • Three dose levels plus controls
  • Dose during organogenesis
    • Rat - gestation days 6-18
    • Rabbit - gestation days 6-18
  • Assess ovarian CLs, implantation sites, resorptions, dead fetuses, gross fetal malformations and development variations

Prenatal and Postnatal Development, including Maternal Function - Segment III

  • Prenatal and Postnatal Development
  • Female rat dosed via the clinical route
  • Three dose levels plus controls
  • Dose Gestation Day 6 through Lactation Day 20
  • Assess maternal toxicity, pregnancy duration, parturition, lactation, implantation, embryo/fetal changes, stillbirths, growth development, behavior and reproductive changes

 

STUDY EXAMPLES

ACUTE TOXICITY STUDY EXAMPLES

Single-Dose (Phase I) and 7-Day Repeat Oral Dose Toxicity (Phase II) Study in Rats (Non-GLP)

 

PHASE I (SINGLE DOSE)

  • Non-GLP
  • There will be three groups of 3/sex/group of Sprague Dawley rats (18 total).
  • Animals will be acclimated for at least 7 days prior to study initiation.
  • Doses will be formulated once prior to each phase by the BASi pharmacy staff.
  • Initially, three groups of 3 rats/sex will be given a single dose at 3 dose levels.  The groups will be observed for 3 days and dose levels for Phase II will be based on the results.
  • Test article will be delivered via oral administration
  • A gross necropsy will be performed on all animals found dead or on animals at the end of the 3 day observation period.

 

PHASE II (7-DAY REPEATED DOSE)

  • There will be three groups of 5/sex Sprague Dawley main study rats and three groups of 6/sex/group TK rats (66 total). 
  • Animals will be acclimated for at least 7 days prior to study initiation.
  • Based on the results of Phase I, the 6 groups will be dosed for 7 consecutive days via oral gavage.
  • Doses will be formulated once prior to each phase by the BASi pharmacy staff.
  • A general physical examination will be performed by a qualified technician once prior to study initiation.
  • Clinical observations will be performed at least once daily for animals during the study.  Mortality observations will be performed twice daily for animals throughout the study.
  • Body weights will be recorded once prior to each dose and at termination.
  • Food consumption will be measured daily.
  • TK plasma samples will be collected from all animals at ~6 timepoints from 3/rats/sex/group on Days 1 and 7 shipped to a Sponsor designated labor for analysis (cost provided separately).  N=216 TK samples
  • Clinical pathology (clinical chemistry and hematology) at necropsy.
  • A gross necropsy will be performed on all animals found dead or on animals at the end of the observation period.  Tissues will be preserved for possible evaluation. 

 

Escalating Dose Range Finding Study (Phase I) and 7-Day Repeat Dose Toxicity and Toxicokinetics (Phase II) Study in Dogs/Monkeys (NonGLP)

 

PHASE I (SINGLE DOSE STUDY)

  • Non-GLP
  • 1 dose group with 1 male and 1 female non-naïve animals (N=2)
  • Animals will be oral dosed
  • The dosing formulations will be prepared by the BASi Evansville staff using a procedure provided by the sponsor.
  • Up to 4 doses at determined doses with a 3 day washout in between each dose. 
  • Clinical observations will be recorded twice daily
  • Body weights will be recorded prior to each dose.

 

PHASE II (7-DAY REPEATED DOSE STUDY)

  • Non-GLP
  • 3 dose group with 2/sex/group non-naïve animals (N=12)
  • Animals will be dosed via oral gavage once daily for 7 days.
  • The dosing formulations will be prepared by the BASi Evansville staff using a procedure provided by the sponsor.
  • Clinical observations will be recorded twice daily.
  • Body weights will be recorded weekly.
  • TK plasma samples will be collected from all animals at ~6 timepoints on Days 1 and 7 and shipped to a Sponsor designated labor for analysis.  Blood samples will be spun down, the plasma separated, frozen and shipped to the BASi Analytical Laboratory or a lab designated by the Sponsor for analysis. N=144 TK samples
  • Blood samples for clinical chemistry and hematology parameters will be collected from each animal at predose and prior to necropsy
  • All animals will be sacrificed and a gross necropsy performed on Day 8.  Tissues will be collected and preserved for possible evaluation.

SUBCHRONIC TOXICITY STUDY EXAMPLES

28-Day GLP Oral Gavage Toxicology Study in Rats with a 4-Week Recovery Period

  • This study will be designed to conform to the FDA Good Laboratory Practice for Nonclinical Studies regulations (CFR 21 Part 58).
  • Rats will be dosed via oral gavage daily for at least 28 days.
  • Four groups of 10/sex/group main study animals with 5/sex/group for recovery (n=120).  An additional 6/sex/group will be assigned to Groups 2-4 for TK collections.  Additional 3/sex in control Group 1 for TK collections. (N=42)  Total of 162 animals on study

 

Group

Dosage
mg/kg

Terminal Sacrifice

Recovery Sacrifice

TK Animals

M

F

M

M

F

F

1

0 (vehicle)

10

10

5

5

3

3

2

TBD

10

10

5

5

6

6

3

TBD

10

10

5

5

6

6

4

TBD

10

10

5

5

6

6

 

  • Physical examinations, including ophthalmology, will be performed by a qualified veterinarian on all study animals prior to initiation.  Ophthalmology will be performed on all main study animals at the end of the dosing and recovery phases.
  • Clinical observations will be performed at least once-daily for main study animals during the study.  Mortality observations will be performed twice daily for all study animals
  • Body weights will be recorded once prior to treatment and at least once weekly during the dosing phase and the recovery period.  Terminal body weights will be recorded for main study animals and recovery animals prior to necropsy, following an overnight fast.
  • Dosing formulations will be prepared weekly.  Samples will be collected at protocol indicated intervals and shipped to the BASi Analytical Laboratory or a lab designated by the Sponsor for analysis. 
  • From TK rats, blood samples will be collected at 6 time points over a 24 hour period from 3/rats/sex/group in Groups 2-4 and at one timepoint from animals in the control group on Days 1 and and near the end of the dosing phase.  The samples will be spun down, the plasma separated, frozen and shipped to the BASi Analytical Laboratory or a lab designated by the Sponsor for analysis (N=228 TK samples). 
  • Terminal blood samples for clinical pathology analyses (hematology, serum chemistry, coagulation – PT & APTT), will be collected from main study animals at their respective necropsy.  At unscheduled necropsies of moribund animals, blood samples will be collected prior to euthanasia, when possible. 
  • Urinalysis near the end of the dosing period and near the end of the recovery period on 5/sex/group.
  • Following at least 28 days of consecutive dosing, the first 10 main study rats/sex/group will be euthanized with carbon dioxide inhalation, exsanguinated and a gross necropsy performed.  The remaining animals will be euthanized following a 4-week recovery period. 
  • A standard list of tissues will be collected and preserved in 10% NBF.
  • A standard list of tissues will be weighed.
  • All collected tissues from Groups 1 and 4, tissues from animals that die or are euthanized early, and any tissues with gross lesions will be embedded in paraffin, processed into blocks, sectioned, mounted on slides, stained, and examined by a veterinary pathologist (ACVP).  Potential target organs identified upon consultation with pathologist, study director and study monitor will be similarly processed and examined from remaining rats at an additional cost.

  .

28-Day Oral GLP Toxicology Study in Dogs/Monkeys with a 4 Week Recovery Period

  • This study will be designed to conform to the FDA Good Laboratory Practice for Nonclinical Studies regulations (CFR 21 Part 58).
  • Animals will be dosed once daily via oral gavage once a day for at least 28 consecutive days.
  •  Four groups of 4/sex/group main study animals + 2/sex/group in the control and high dose groups for recovery.  N= 40

 

Group

Dosage
mg/kg

Terminal Sacrifice

Recovery Sacrifice

M

F

M

F

1

0 (vehicle)

4

4

2

2

2

TBD

4

4

-

-

3

TBD

4

4

-

-

4

TBD

4

4

2

2

 

  • Dosing formulations will be prepared weekly.  Samples will be collected at protocol indicated intervals and shipped to the BASi Analytical Laboratory or a lab designated by the Sponsor for analysis. 
  • Blood samples will be collected at 8 time points from Groups 2, 3, & 4 and one timepoint from the Control Group on Day 1 and near the end of the dosing phase. The samples will be spun down, the plasma separated, frozen and shipped to the BASi Analytical Laboratory or a lab designated by the Sponsor for analysis
  • Physical examinations, including ophthalmology, will be performed by a qualified veterinarian on all study animals prior to initiation.  Ophthalmology will be performed on all study animals prior to initiation, during Week 26 and near the end of the dosing and recovery phases.
  • ECGs will be recorded for all study animals prior to initiation (2 pretest ECGs), on Days 1 near or at the end of the dosing and recovery phases. 
  • Clinical observations will be performed at least once-daily for all study animals during the study.  Mortality observations will be performed twice daily for all study animals
  • Body weights will be recorded once prior to treatment and at least once weekly during the dosing phase and the recovery phase.  Terminal body weights will be recorded for animals prior to their respective necropsy, following an overnight fast.
  • Daily food consumption estimates
  • Hematology, serum chemistry, and coags once pretest and near the end of the dosing and recovery phases.
  • Urinalysis once pretest, at the end of the dosing and recovery phases, if available.
  • Following at least 28 days of dosing, the first 4 animals/sex/group will be euthanized with sodium pentobarbital, exsanguinated and a gross necropsy performed.  The remaining animals will be allowed to recover for at least 4 weeks. 
  • A standard list of tissues will be collected and preserved in 10% NBF.
  • A standard list of tissues will be weighed.
  • All collected tissues will be trimmed, embedded in paraffin, sectioned, slides prepared and examined by a veterinary pathologist (ACVP).