[1,1':3',1''-Terphenyl]-2'-ol, 5',5''''-(2,2,2-trifluoro-1-methylethylidene)bis-

Repeated dose toxicity: oral

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: study according to GLP and OECD guideline

Cross-reference

Reason / purpose for cross-reference:

reference to same study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2015

Report date:

2015

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 422 (Combined Repeated Dose Toxicity Study with the Reproduction / Developmental Toxicity Screening Test)

Test guideline 2

Qualifier:

according to guideline

Guideline:

other: US EPA OPPTS 870.3650, Combined Repeated Dose Toxicity Study with the Reproduction/Developmental Toxicity Screening Test

Test guideline 3

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 407 (Repeated Dose 28-Day Oral Toxicity Study in Rodents)

Test guideline 4

Qualifier:

equivalent or similar to guideline

Guideline:

EU Method B.7 (Repeated Dose (28 Days) Toxicity (Oral))

Test guideline 5

Qualifier:

equivalent or similar to guideline

Guideline:

other: US EPA OPPTS 870.3050, Repeated dose 28-day oral toxicity study in rodents

GLP compliance:

yes (incl. QA statement)

Remarks:

testing lab.

Limit test:

no

Test material

Test material information

Constituent 1

Reference substance name:

3-phenyl-5-(1,1,1-trifluoro-2-{6-hydroxy-5-phenyl-[1,1'-biphenyl]-3-yl}propan-2-yl)-[1,1'-biphenyl]-2-ol

EC Number:

610-104-3

Cas Number:

43100-47-6

Molecular formula:

C39 H29 F3 O2

IUPAC Name:

3-phenyl-5-(1,1,1-trifluoro-2-{6-hydroxy-5-phenyl-[1,1'-biphenyl]-3-yl}propan-2-yl)-[1,1'-biphenyl]-2-ol

Details on test material:

- Name of test material (as cited in study report): Ligand TFME-DPP
- Physical state: White solid
- Analytical purity: 94.6% (quantitative 1H-NMR-spectroscopy)
- Lot/batch No.: 01829-00216
- Expiration date of the lot/batch: 30 June 2016
- Storage condition of test material: at room temperature

Test animals

Species:

rat

Strain:

Wistar

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River Deutschland, Sulzfeld, Germany
- Age at study initiation: males approx. 11 weeks, females approx. 12 weeks
- Housing: groups of 5 animals of the same sex (pre-mating), females were caged with males on a one-to-one basis (mating), males were housed in home cage at a maximum of 5/cage, females were individually housed (post-mating)
- Diet: ad libitum
- Water: tap water ad libitum
- Acclimation period: at least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 18-24
- Humidity (%):40-70
- Air changes (per hr): at least 10
- Photoperiod (hrs dark / hrs light): 12/12

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

polyethylene glycol

Remarks:

400, specific gravity 1.125

Details on oral exposure:

PREPARATION OF DOSING SOLUTIONS:
Formulations (w/w) were prepared daily within 6 hours prior to dosing and were homogenized to a visually acceptable level.

VEHICLE
- Justification for use and choice of vehicle (if other than water): Based on trial formulations performed at WIL Research Europe
- Amount of vehicle (if gavage): 5ml/kg bw

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

Analyses were conducted on a single occasion during the treatment phase (18 February 2015). The concentrations analyzed in the formulations were in agreement with the target concentrations (i.e. mean accuracies between 85% and 115%). The formulations of the high and low dose groups were assessed for homogeneity and were found to be homogenous. Formulations were stable at room temperature for at least 6 hours.

Duration of treatment / exposure:

males: 31 days (2 weeks prior to mating, during mating, and up to the day before necropsy)
females: 40-53 days (2 weeks prior to mating, during mating, during post-coitum, and during at least 4 days of lactation); exception: 3 females in group 4 were not dosed on day 21 or 22 post coitum since they were littering at the time of dosing.

Frequency of treatment:

daily, approximately at the same time each day

Doses / concentrations

Remarks:

Doses / Concentrations:
100, 300, 1000 mg/kg bw/d
Basis:

No. of animals per sex per dose:

10

Control animals:

yes, concurrent vehicle

Details on study design:

- Dose selection rationale: based on a 14-day dose range finding study, up to 1000 mg/kg bw/d were tested without dose limiting effects

Examinations

Observations and examinations performed and frequency:

CAGE SIDE OBSERVATIONS: Yes
- Time schedule: at least twice daily

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: daily

BODY WEIGHT: Yes
- Time schedule for examinations: on the first day of exposure and weekly thereafter. Mated females were weighed on Days 0, 4, 7, 11, 14, 17 and 20 post-coitum and during lactation on Days 1 and 4.

FOOD CONSUMPTION :
Weekly, except for males and females which were housed together for mating and for females without evidence of mating. Food consumption of mated females was measured on Days 0, 4, 7, 11, 14, 17 and 20 post-coitum and on Days 1 and 4 of lactation.

WATER CONSUMPTION: No

OPHTHALMOSCOPIC EXAMINATION: No

HAEMATOLOGY: Yes
- Time schedule for collection of blood: on the day of scheduled necropsy
- Anaesthetic used for blood collection: Yes (isoflurane)
- Animals fasted: Yes (overnight)
- How many animals: 5 animals/sex/group
- Parameters examined: white blood cells, differential leukocyte count, red blood cells, reticulocytes, red blood cell distribution width, haemoglobin, haematocrit, MCV, MCH, MCHC, platelets, prothrombin time, activated partial thromboplastin time

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: on the day of scheduled necropsy
- Animals fasted: Yes
- How many animals: 5/sex/group
- Parameters examined: ALAT, ASAT, ALP, total protein, albumin, total bilirubin, urea, creatinine, glucose, cholesterol, sodium, potassium, chloride, calcium, phosphate, bile acids

URINALYSIS: No

NEUROBEHAVIOURAL EXAMINATION: Yes
- Time schedule for examinations: males during week 4 of treatment, females from lactation day 4 onward (before blood sampling)
- Dose groups that were examined: 5 animals/sex/group
- Battery of functions tested: sensory activity / grip strength / motor activity

OTHER:
- pup mortality was determined on day 1 of lactation and daily thereafter.
- cliinical signs for pups were recorded at least once daily
- body weight of live pups was determined on Days 1 and 4 of lactation
- sex was determined for all pups on days 1 and 4 of lactation

Sacrifice and pathology:

GROSS PATHOLOGY: Yes
- males at completion of mating period (at least 28 days of dose administration), females which deliver: lactation days 5-7, females which fail to deliver: post-coitum days 25-27
- organ weights: 5 animals/sex/group, adrenal glands, brain, epididymides (all males), heart, kidneys, liver, ovaries, spleen, testes (all males), thymus, uterus (including cervix), prostate (weighed when fixed for at least 24 hours), seminal vesicles including coagulation glands (weighed when fixed for at least 24 hours), thyroid including parathyroid

HISTOPATHOLOGY: Yes
- The following organs were preserved for all animals, and organs and tissues of the selected 5 animals/sex of Groups 1 and 4 were analyzed except for those organs listed in parentheses: adrenal glands, (aorta), brain, caecum,cervix, clitoral gland, colon, coagulation gland, duodenum, epididymides, eyes, mammary gland area, femur including joint, heart, ileum, jejunum, kidneys, liver, lung, lymph nodes, ovaries, (pancreas), peyer's patches, (esophagus), pituitary gland, preputial gland, prostate gland, rectum, (salivary glands), sciatic nerve, seminal vesicles, sceletal muscle, (skin), spinal cord, spleen, sternum, stomach, testes, thymus, thyroid, (tongue), trachea, urinary bladder, uterus, vagina, all gross lesions.
- The additional slides of the testes of all males of Groups 1 and 4 and all males that failed to sire to examine staging of spermatogenesis and histopathology of interstitial cell structure.
- The liver of the selected males of Groups 2 and 3 based on suspected treatment-related microscopic findings in this organ.
- All gross lesions of all animals (all dose groups).
- The reproductive organs (cervix, clitoral gland, coagulation gland, epididymides, ovaries, preputial gland, prostate gland, seminal vesicles, testes, uterus, and viagina) of all animals of Groups 1 and 4 and all males that failed to sire and all females that failed to deliver healthy pups.

Statistics:

The following statistical methods were used to analyse the data:
- If the variables could be assumed to follow a normal distribution, the Dunnett-test (Ref. 2; many-to-one t-test) based on a pooled variance estimate was applied for the comparison of the treated groups and the control groups for each sex.
- The Steel-test (Ref. 3; many-to-one rank test) was applied if the data could not be assumed to follow a normal distribution.
- The Fisher Exact-test (Ref. 4) was applied to frequency data.
- The Kruskal-Wallis nonparametric ANOVA test (Ref. 5) was applied to motor activity data to determine intergroup differences.

All tests were two-sided and in all cases p < 0.05 was accepted as the lowest level of significance.

Results and discussion

Results of examinations

Clinical signs:

no effects observed

Mortality:

no mortality observed

Body weight and weight changes:

no effects observed

Food consumption and compound intake (if feeding study):

no effects observed

Food efficiency:

not examined

Water consumption and compound intake (if drinking water study):

not examined

Ophthalmological findings:

not examined

Haematological findings:

no effects observed

Clinical biochemistry findings:

no effects observed

Urinalysis findings:

not examined

Behaviour (functional findings):

no effects observed

Organ weight findings including organ / body weight ratios:

effects observed, treatment-related

Description (incidence and severity):

Liver weight was increased in males in dose-dependent manner from 100 mg/kg bw/d.

Gross pathological findings:

no effects observed

Histopathological findings: non-neoplastic:

effects observed, treatment-related

Description (incidence and severity):

Minimal hepatocellular hypertrophy was detected in 2/5 males at 300 mg/kg bw/d and 4/5 males in 1000 mg/kg bw/d

Histopathological findings: neoplastic:

no effects observed

Effect levels

Dose descriptor:

NOAEL

Effect level:

1 000 mg/kg bw/day (actual dose received)

Based on:

test mat.

Sex:

male/female

Target system / organ toxicity

Critical effects observed:

not specified