4-Pyrimidineethanol, 6-chloro-.alpha.-(2,4-difluorophenyl)-5-fluoro-.beta.-methyl-.alpha.-(1H-1,2,4-triazol-1-ylmethyl)-, hydrochloride (1:1)

Repeated dose toxicity: oral

Administrative data

Endpoint:

short-term repeated dose toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

From 19 May 1999 to 17 June 1999

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Study run to a method comparable with current guidelines and to GLP.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1999

Report date:

1999

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

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

Deviations:

no

GLP compliance:

yes

Limit test:

no

Test material

Test material information

Constituent 1

Reference substance name:

3-(6-chloro-5-fluoropyrimidin-4-yl)-2-(2,4-difluorophenyl)-1-(1H-1,2,4-triazol-1-yl)butan-2-ol hydrochloride

EC Number:

928-729-8

Cas Number:

188416-20-8

Molecular formula:

C16H13ClF3N5O.HCl

IUPAC Name:

3-(6-chloro-5-fluoropyrimidin-4-yl)-2-(2,4-difluorophenyl)-1-(1H-1,2,4-triazol-1-yl)butan-2-ol hydrochloride

Details on test material:

Lot/batch No.:109496/F/33
Purity: not specified

Test animals

Species:

rat

Strain:

Crj: CD(SD)

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source:Charles River(UK) Limited, Margate, Kent, England
- Age at study initiation:34 to 38 days of age
- Weight at study initiation:The male animals used on the study weighed 136-157 g on the day that treatment commenced; the females were within the bodyweight range 124-139 g at this time.
- Fasting period before study:The animals were allowed free access, except overnight before routine blood sampling, to an expanded rodent diet, Rat and Mouse No.1 Matentiance Diet (Special Diets Services Limited, Witham, Essex, England).
- Housing:yes, a stainless steel body with a stainless steel mesh lid and floor
- Diet (e.g. ad libitum):This diet contained no added antibiotic or chemotherapeutic or prophylactic agent.
- Water (e.g. ad libitum):Water taken from the public supply was freely available, via polycarbonate bottles fitted with sipper tubes.
- Acclimation period:yes, eight days

ENVIRONMENTAL CONDITIONS
- Temperature (°C):21°C for temperature (acceptable range 19-25°C)
- Humidity (%):55% for relative humidity(acceptable range 40-70%)
- Air changes (per hr):
- Photoperiod (hrs dark / hrs light):a 12-hour light: 12-hour dark cycle

IN-LIFE DATES: From:1999-5-19 To:1999-6-17

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

corn oil

Details on oral exposure:

DIET PREPARATION
- Rate of preparation of diet (frequency):Weighed amounts of diet were provided at intervals during each week.
- Mixing appropriate amounts with (Type of food):
- Storage temperature of food:

VEHICLE
- Justification for use and choice of vehicle (if other than water):Uk-103,446-01 was homogenous and stable in corn oil for up to 15 days when stored at 4°C.
- Concentration in vehicle:at 1.0 and 100 mg/mL
- Amount of vehicle (if gavage):at a constant volume-dosage of 5mL/kg bodyweight
- Lot/batch no. (if required):
- Purity:

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

HPLC

Duration of treatment / exposure:

28 days

Frequency of treatment:

once each day, seven days per week.

Doses / concentrations

Remarks:

Doses / Concentrations:
15,150 and 500 mg/kg/day
Basis:
actual ingested

No. of animals per sex per dose:

5 male and 5 female

Control animals:

yes, concurrent vehicle

Examinations

Observations and examinations performed and frequency:

CAGE SIDE OBSERVATIONS: Yes
- Time schedule:Immediately before dosing. Immediately after dosing on return of the animal to its cage. On completion of dosing of each group. Between one and two hours after completion of dosing of all groups. As late as possible in the working day.
- Cage side observations checked in table [No.?] were included.

BODY WEIGHT: Yes
- Time schedule for examinations:Each animal was weighed during the acclimatisation period, on the day that treatment commenced, twice weekly throughout the treatment period and before necropsy.

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: No. Weighed amounts of diet were provided at intervals during each week to each cage. At the end of each treatment week the weight of uneaten food was recorded. The uneaten diet may have been included in that returned to the cage, after appropriate measurement.
- Compound intake calculated as time-weighted averages from the consumption and body weight gain data: No data

FOOD EFFICIENCY:
- Body weight gain in kg/food consumption in kg per unit time X 100 calculated as time-weighted averages from the consumption and body weight gain data: Yes

HAEMATOLOGY: Yes
- Time schedule for collection of blood:On Day 29 of treatment, blood samples were obtained from all animals.
- Anaesthetic used for blood collection: No data
- Animals fasted: Yes
- How many animals:40
- Parameters checked in table [No.?] were examined.

Sacrifice and pathology:

GROSS PATHOLOGY: Yes
HISTOPATHOLOGY: Yes

Results and discussion

Results of examinations

Clinical signs:

effects observed, treatment-related

Mortality:

mortality observed, treatment-related

Body weight and weight changes:

effects observed, treatment-related

Food consumption and compound intake (if feeding study):

effects observed, treatment-related

Food efficiency:

no effects observed

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

no effects observed

Ophthalmological findings:

not specified

Haematological findings:

no effects observed

Clinical biochemistry findings:

effects observed, treatment-related

Urinalysis findings:

not specified

Behaviour (functional findings):

no effects observed

Organ weight findings including organ / body weight ratios:

effects observed, treatment-related

Gross pathological findings:

no effects observed

Histopathological findings: non-neoplastic:

no effects observed

Histopathological findings: neoplastic:

no effects observed

Details on results:

No animals died.
Salivation after dose administration was observed from Day2 onwards in males and females receiving 150 or 500 mg/kg/day, with a few animals receiving 15 mg/kg/day also being affected transiently.
Bodyweight gain and food intake was high, compared with the Controls, in females receiving 500 mg/kg/day and bodyweight gain was high in females receiving 150 mg/kg/day. The bodyweight gain and food intake of males was unaffected. Food conversion efficiency in both sexes was unaffected by treatment.
No clear effect of treatment was identified at the functional observation battery investigations.
There were no treatment-related haematological changes after four weeks of treatment.
Biochemical changes in the plasma that were attributed to treatment comprised: high alkaline phosphatase and alanine and aspartate amino-transferase activities and high urea concentrations in males receiving 500mg/kg/day; high creatinine and phosphorus concentrations in males and females receiving 500mg/kg/day; low glucose concentrations in females receiving 500mg/kg/day; slightly high calcium concentrations in females receiving 150 or 500 mg/kg/day.
After four weeks of treatment liver weights were slightly high in females receiving 500mg/kg/day and spleen weights were slightly low in males at this dosage.
There were no treatment-related macroscopic or histopathological changes.

Effect levels

1

Dose descriptor:

NOAEL

Effect level:

ca. 500 mg/kg bw/day (actual dose received)

Based on:

act. ingr.

Sex:

male/female

Basis for effect level:

other: no findings at toxicological significance at highest dose level

2

Dose descriptor:

LOEL

Effect level:

ca. 15 mg/kg bw/day (actual dose received)

Based on:

act. ingr.

Sex:

male/female

Basis for effect level:

other: overall effects clinical signs; body weight; food consumption; food efficiency; water consumption and compound intake; haematology; gross pathology; organ weights; histopathology

Target system / organ toxicity

Critical effects observed:

not specified

Applicant's summary and conclusion

Conclusions:

It is concluded that oral administration of this substance to CD rats at dosages up 500mg/kg/day for four weeks was well-tolerated, producing no toxicologically significant finding. This study provided no evidence for any neurotoxic potential. An adaptive response by the liver was indicated at the high dosage. There were no changes at 500mg/kg/day that were considered to be of any toxicological significance and this dosage is, therefore, considered to be the no-observed-adverse-effect(NOAEL) in this study.The lowest observed effect level(LOEL) was considered to be 15 mg/kg/day.