Registration Dossier

Administrative data

Endpoint:
short-term repeated dose toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: OECD Guideline Study according to GLP

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2013
Report Date:
2013

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 407 (Repeated Dose 28-Day Oral Toxicity in Rodents)
Version / remarks:
adopted 03 Oct 2008
GLP compliance:
yes (incl. certificate)
Remarks:
BASF SE Experimental Toxicology and Ecology 67056 Ludwigshafen, Germany
Limit test:
no

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
- Name of test material (as cited in study report): L4-Ligand
- Physical state: solid/ white
- Analytical purity: 97.0 g/100 g (tolerance ± 0.3%)
- Lot/batch No.: 0005473663
- Storage condition of test material: ambient (RT); under N2 ; protect against humidity

Test animals

Species:
rat
Strain:
Wistar
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Charles River Laboratories, Research Models and Services, Sulzfeld, Germany
- Age at study initiation: 42 ± 1 day(s)
- Weight at study initiation: mean: male: 156.3 g; female: 128.9 g
- Housing: 5 animals per cage
- Diet: ad libitum
- Water: ad libitum
- Acclimation period: 7 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20 - 24
- Humidity (%): 30 - 70
- Air changes (per hr): 15
- Photoperiod (hrs dark / hrs light): 12/12

Administration / exposure

Route of administration:
oral: feed
Details on oral exposure:
DIET PREPARATION
- Rate of preparation of diet: in a frequency for which stability in the carrier was demonstrated (8 days stored deep frozen followed by 1 day at room temperature)
- Mixing appropriate amounts with (Type of food): ground Kliba maintenance diet mouse/rat “GLP” meal
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
All values were in the range of 90 - 110% of the target concentration.
Duration of treatment / exposure:
4 weeks
Frequency of treatment:
daily
Doses / concentrationsopen allclose all
Remarks:
Doses / Concentrations:
0, 1200, 3600 and 12000 ppm
Basis:
nominal in diet
Remarks:
Doses / Concentrations:
0, 101.8, 300.6, 994.7 mg/kg bw/day for males
Basis:
actual ingested
Remarks:
Doses / Concentrations:
0, 104.0, 300.4, 1018.4 mg/kg bw/day for females
Basis:
actual ingested
No. of animals per sex per dose:
5
Control animals:
yes, plain diet

Examinations

Observations and examinations performed and frequency:
CAGE SIDE OBSERVATIONS: Yes
- Time schedule: at least once a day

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: prior the start of the administration period and subsequently once weekly (in the morning) thereafter

BODY WEIGHT: Yes
- Time schedule for examinations: once weekly

FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study):
- Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: Yes
- Compound intake calculated as time-weighted averages from the consumption and body weight gain data: Yes

HAEMATOLOGY: Yes
- Time schedule for collection of blood: last day of study
- Anaesthetic used for blood collection: Yes (isoflurane)
- Animals fasted: Yes
- How many animals: all
- Parameters checked: Leukocyte count (WBC), Erythrocyte count (RBC), Hemoglobin (HGB), Hematocrit (HCT), Mean corpuscular volume (MCV), Mean corpuscular hemoglobin (MCH), Mean corpuscular hemoglobin concentration (MCHC), Platelet count (PLT), Differential blood count, Reticulocytes, Prothrombin time (Hepato Quick’s test) (HQT)

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: last day of study
- Animals fasted: Yes
- How many animals: all
- Parameters checked: Alanine aminotransferase (ALT), Aspartate aminotransferase (AST), Alkaline phosphatase (ALP), γ-Glutamyltransferase (GGT), Sodium (NA), Potassium (K), Chloride (Cl), Inorganic phosphate (INP), Calcium (Ca), Urea, Creatinine, Glucose, Total bilirubin, Total protein, Albumin, Globulins, Triglycerides, Cholesterol, Bile acid

URINALYSIS: Yes
- Time schedule for collection of urine: last day of study
- Metabolism cages used for collection of urine: Yes
- Animals fasted: Yes
- Parameters checked: pH, protein, glucose, ketones, urobilinogen, bilirubin, blood, specific gravity, sediment, color, turbidity, volume

NEUROBEHAVIOURAL EXAMINATION: Yes
- Time schedule for examinations: prior the start of the administration period and subsequently once weekly (in the morning) thereafter
- Dose groups that were examined: all
- Battery of functions tested: functional observational battery and measurement of motor activity
Sacrifice and pathology:
GROSS PATHOLOGY: Yes
HISTOPATHOLOGY: Yes

Results and discussion

Results of examinations

Details on results:
CLINICAL SIGNS AND MORTALITY: No treatment-related signs of general systemic toxicity were observed. No animals died or were sacrificed moribund prematurely in the present study.

BODY WEIGHT AND WEIGHT GAIN: No changes in body weight parameters were observed in all test groups.

FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study): No test substance-related effects on food consumption were obtained. All detected values were within the normal range typical for rats of this strain and age.

HAEMATOLOGY: No treatment-related changes of hematological parameters were observed. At the end of the study in males of test group 1 (1200 ppm) hematocrit values were higher compared to controls, but alterations were not dose dependent. Therefore, this change was regarded as incidental and not treatment-related.

CLINICAL CHEMISTRY: No treatment-related changes of clinico-chemical parameters were observed.

URINALYSIS: No treatment-related changes among urinalysis parameters were observed.

NEUROBEHAVIOUR: There were no test substance-related findings in male and female animals of all test groups.

ORGAN WEIGHTS: When compared to control group 0 (set to 100%), the mean absolute weights of thryroid glands were significantly increased in test group 1 (1200 ppm; 128% )and test group 3 (12000 ppm; 122%). All other mean absolute weight parameters did not show significant differences when compared to the control group 0.
When compared to control group 0 (set to 100%), the mean relative weights of following organs were significantly increased: liver of test group 1 (1200 ppm; 107%) and test group 3 (12000 ppm; 112%); thyroid glands of test group 3 (12000 ppm; 121%).
There was no dose-response relationship in absolute and relative thyroid weights nor was there a histopathological correlate for the changes in thyroid weights, therefore these effects were regarded to be incidental and not related to treatment. The liver weight increase in test group 3 female animals (12000 ppm) was likely treatment–related as fatty change was detected histopathologically. The minimally increased liver weights in test group 1 females (1200 ppm) was not considered to be related to treatment as there was no histopathological correlate and there was no clear dose response.

GROSS PATHOLOGY: All findings occurred either individually or were biologically equally distributed over control and treatment groups. They were considered to be incidental or spontaneous in origin and without any relation to treatment.

HISTOPATHOLOGY: NON-NEOPLASTIC: The liver in female animals of groups 2 and 3 showed periportal fatty change. Most other findings occurred either individually or were biologically equally distributed over control and treatment groups. Sperm granulomas of the epididymis occurred only in treated animals in this study (1/5 in test group 2 [3600 ppm] and 2/5 in test group 3 [12000 ppm]). This finding is, however, known to be a common finding in young rats (Creasey D. 2012 Chapter 9 Reproduction of the rat, mouse, dog, non- human primate and minipig. In: Background Lesions in Laboratory Animals. A color atlas. EF McInnes (Ed) Saunders Elsevier , pp. 103). This finding was considered to be incidental or spontaneous in origin and without any relation to treatment.

Effect levels

open allclose all
Dose descriptor:
NOAEL
Effect level:
12 000 ppm
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: highest dose tested
Dose descriptor:
NOAEL
Effect level:
994.7 mg/kg bw/day (actual dose received)
Based on:
test mat.
Sex:
male
Basis for effect level:
other: highest dose tested
Dose descriptor:
NOAEL
Effect level:
1 018.4 mg/kg bw/day (actual dose received)
Based on:
test mat.
Sex:
female
Basis for effect level:
other: highest dose tested

Target system / organ toxicity

Critical effects observed:
not specified

Applicant's summary and conclusion