GLP 14 Day Repeat Dosing Toxicology in Rats
This GLP study design is used to determine the toxicity of a test article in male and female rats after repeat dosing for 14 days followed by a 7 day recovery period
Catalog number EA609
Required from Customer
- Test article in a form appropriate for dosing; see Note 1
- Instructions for dosing vehicle preparation
- Dose, dose route, and test article stability information
- Test article Certificate of Analysis (COA)
- Master Study Protocol (MSP) and Bioanalytical Study Plan (BSP) must be written and approved prior to study initiation
Deliverables
- Toxicology report
- No-adverse event limit (NOAEL)
- In-life observation: mortality, morbidity, clinical signs, and body weight
- Clinical pathology, clinical chemistry, and necropsy results
- If optional toxicokinetics (TK) analysis is included:
- Table of measured concentration of test article in each sample
- Table of TK parameters (AUC, Cmax, and Tmax) for each dose route
- Half-life (t1/2), clearance and volume of distribution after IV adminstration
- Bioanalytical method and validation data
- Raw analytical data
- Representative chromatograms and calibration curves
Substrate
- Test article in an appropriate dosing vehicle
Study System
- Conscious, male and female Sprague-Dawley rats weighing between 200 and 400 grams following ≥7 days of pre-study acclimation. Water is available ad libitum.
Study Conditions
- Administer test article in parallel to groups of rats (total of 136 animals) according to the design in the Table on page 2
- Each treatment by single oral gavage , IV, SC, or IP dose
- Treat each group once daily for 14 days
- Allow recover animals to recover for 7 days
- In-life observations and terminal investigations; see Note 2
- Optional TK (satellite TK animals only); see Note 6
Study QC and QA
- 5/8 of the calibration standards must be within ±15% of the theoretical values (±20% at the LLOQ)
- Two-thirds of the batch QCs must be accurate to within of nominal, and at least one QC must pass at each level in order to accept the run
- Incurred sample re-analysis will be performed to confirm bioanalytical assay reproducibility
- QA Unit performs in-study and post-study audits
Notes
- Test article requirements for development and validation of the bioanalytical method (in addition to the amount needed for the in vivo study):
- Small molecule: ~70 mg of test article in powder form and ~30 mg of deuterated internal standard in powder form for bioanalysis
- Biologic or other test article: Amount TBD
- If required, Absorption Systems can determine a suitable dosing vehicle, at an additional charge, and prepare it for the study. Alternatively, Absorption Systems can prepare the dosing vehicle according to instructions provided by the Customer.
- In-life observations and terminal investigations
- Mortality and morbity: twice daily
- Clinical signs: daily and 1 hour post-dose on Day 1
- Body weight: weekly (including pre-dose) and at termination
- Clinical pathology: hematology, coagulation and clinical chemistry at termination on all main and recovery animals
- Necropsy: Full gross necropsy with macroscopic examination on all animals and preservation of a full tissue list
- Organ weights: All organ weights from animals examined at necropsy
- Histopathology: All tissues from high dose and control groups for main study animals and recovery animals will be brought to slide, H&E stained, and examined by a board-certified pathologist. If abnormalities are seen, the next lower dose group will be examined for pathology, a process that will continue until no abnormalities are seen or all groups have been examined. Price does not include histopathology for additional groups (after high dose and control).
- The standard anticoagulant is sodium heparin. The customer can specify that another anticoagulant be used.
- The test article must be a small molecule for Absorption Systems to perform plasma sample analysis. Otherwise, plasma samples will be shipped to the Customer for analysis.
- Optional TK includes:
- Sampling at 6 time points each (up to 24 hours post-dosing) on Day 1 and Day 14
- Prepare plasma from blood samples and store at -70°C prior to analysis
- Terminate TK animals without further investigation after last bleed
- Determine the concentrations of test article in dosing solution and incurred samples using a generic LC/MS/MS method with a minimum 5 point calibration curve
- Use non-compartmental analysis to determine TK parameters
Group |
Treatment |
Main Study |
Recovery | TK (optional) |
|||
Male |
Female | Male |
Female | Male |
Female | ||
1 | Vehicle | 10 | 10 | 5 | 5 | 0 | 0 |
2 | Low Dose | 10 | 10 | 0 | 0 | 6 | 6 |
3 | Mid Dose | 10 | 10 | 0 | 0 | 6 | 6 |
4 | High Dose | 10 | 10 | 5 | 5 | 6 | 6 |
Options
- The customer must specify:
• the dose route—oral, IV, SC, or IPthe doses (mg/kg)
• the formulation instructions, if applicable
• the sampling time points for TK, if conducted
• which organs to be collected or weighed after termination - The customer can request:
• collection and bioanalysis of plasma samples for TK analysis
• that Absorption Systems determine a suitable dosing vehicle
• that Absorption Systems prepare the dosing vehicle
• an anticoagulant other than sodium heparin
• histopathology on selected organs