Bicyclo[2.2.1]heptane-1-methanesulfonic acid, 7,7-dimethyl-2-oxo-, (1R,4S)-, compd. with rel-(.alpha.R,.beta.S)-.alpha.-(2,4-difluorophenyl)-5-fluoro-.beta.-methyl-.alpha.-(1H-1,2,4-triazol-1-ylmethyl)-4-pyrimidineethanol (1:1)

Endpoint summary

Description of key information

Acute oral toxicity:
An acute oral toxicity study (HLS, 1994) was available which is key study. This study showed that the oral LD50 value of test substance is between 50 to 500 mg/kg body weight.
Acute dermal toxicity:
An acute dermal toxicity study (McRae, 1998) was available which is key study. This study showed that the oral LD50 value of test substance is established to exceed 2000 mg/kg body weight.

Key value for chemical safety assessment

Acute toxicity: via oral route

Link to relevant study records

Reference

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

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

Qualifier:

according to guideline

Guideline:

EU Method B.1 bis (Acute Oral Toxicity - Fixed Dose Procedure)

Deviations:

no

GLP compliance:

yes

Test type:

fixed dose procedure

Limit test:

no

Species:

rat

Strain:

Sprague-Dawley

Sex:

male/female

Route of administration:

oral: gavage

Vehicle:

other: 1% w/v aqueous methylcellulose

Doses:

A group of ten fasted rats(five males and five femals) was given a single dose by oral gavage of the test substance, formulated in 1% w/v aqueous methylcellulose and administered at an initial dose level of 500 mg/kg bodyweight. A further group of five males and five females rats was dosed at 50 mg/kg bodyweight to determine the discriminating dosage of the test material.

No. of animals per sex per dose:

Preliminary sighting study: 2 (female)
Main study: 5 (male)
Main study: 5 (female)

Control animals:

no

Details on study design:

A group of ten fasted rats(five males and five femals) was given a single dose by oral gavage of the test substance, formulated in 1% w/v aqueous methylcellulose and administered at an initial dose level of 500 mg/kg bodyweight. This dose level was chosen on the basis of preliminary study results and in compliance with the study guideling. A further group of five males and five females rats was dosed at 50 mg/kg bodyweight to determine the discriminating dosage of the test material.

Preliminary study:

Species/strain: Rat (Sprague-Dawley)
2000 mg/kg bw: Evident toxicity: Y; Mortality: Y
500 mg/kg bw: Evident toxicity: Y; Mortality: N
Observations:
These results were used to set the starting dose for the main study.

Sex:

male/female

Dose descriptor:

discriminating dose

Effect level:

50 mg/kg bw

Based on:

act. ingr.

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 50 - < 500 mg/kg bw

Based on:

act. ingr.

Mortality:

Three males and all females at 500 mg/kg died during the study. All deaths occurred within 4 days of dosing.
Female 500 mg/kg bw (fixed dose further) No. with evident toxicity: 5; No. of deaths: 5; No. of animals used: 5
Male 500 mg/kg bw (fixed dose further) No. with evident toxicity: 5; No. of deaths: 3; No. of animals used: 5
Female 50 mg/kg bw (fixed dose further) No. with evident toxicity: 5; No. of deaths: 0; No. of animals used: 5
Male 50 mg/kg bw (fixed dose further) No. with evident toxicity: 5; No. of deaths: 0; No. of animals used: 5

Clinical signs:

other: Signs of toxicity: Clinical signs of reaction to treatment included piloerection and hunched posture, seen in all rats at both dosages. In addition, waddling gait, lethargy, decreased respiration, partially closed eyelids, pallor of the extremities, calo

Gross pathology:

Effects on organs:
Macroscopic examination of surviving rats killed on Day 15 revealed no abnormalities.

Other findings:

Three males and all females at 500 mg/kg died during the study. All deaths occurred within four days of dosing.

Interpretation of results:

harmful

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

LD50 is between 50 and 500 mg/kg bw/day.

Endpoint conclusion

Endpoint conclusion:

adverse effect observed

Dose descriptor:

discriminating dose

Value:

50 mg/kg bw

Quality of whole database:

1 (reliable without restriction)

Acute toxicity: via dermal route

Link to relevant study records

Reference

Endpoint:

acute toxicity: dermal

Type of information:

experimental study

Adequacy of study:

key study

Study period:

From 28 January to 11 February 1997

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

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

Qualifier:

according to guideline

Guideline:

EU Method B.3 (Acute Toxicity (Dermal))

Deviations:

no

GLP compliance:

yes

Test type:

standard acute method

Limit test:

yes

Species:

rat

Strain:

Sprague-Dawley

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Harlan U.K.Ltd., Bicester, Oxon, England
- Age at study initiation: Eight to ten weeks of age
- Weight at study initiation: 220 to 250 g
- Fasting period before study:
- Housing: They were housed individually in metal cages with wire mesh floors
- Diet: A standard laboratory rodent diet
- Water: ad libitum
- Acclimation period: A minimum period of five days prior to the start of the study

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19-21ºC
- Humidity (%): 32-52% RH
- Air changes (per hr): Maintained at 10 to 15 air changes per hour
- Photoperiod (hrs dark / hrs light): 12 hours of artificial light (0700-1900 hours) in 24-hour period

IN-LIFE DATES: From 28 January To 11 February 1997

Type of coverage:

occlusive

Vehicle:

other: aqueous methylcellulose

Details on dermal exposure:

TEST SITE
- Area of exposure: Approximately 50 mm * 50 mm
- % coverage: Approximately 10% of the total body surface area
- Type of wrap if used: Covered with porous gauze held in place with a non irritating dressing, and further covered by a waterproof dressing encircled firmly around the trunk of the animal

REMOVAL OF TEST SUBSTANCE
- Washing (if done): Washed with warm water (30 to 40ºC )
- Time after start of exposure: 24 hours

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 2000 mg/kg bodyweight
- Concentration (if solution): A maximum practical concentration of 66.67% w/v in 1% w/v aqueous methylcellulose
- Constant volume or concentration used: no


VEHICLE
- Amount(s) applied (volume or weight with unit): 3 ml/kg bodyweight
- Concentration (if solution): 1% w/v aqueous methylcellulose
- Lot/batch no. (if required):
- Purity:

Duration of exposure:

24h

Doses:

2000 mg/kg bodyweight

No. of animals per sex per dose:

a group of ten rats (five males and five females)

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: Once in the morning and again at the end of the experimental day (with the exception of Day 15-morning only)
- Necropsy of survivors performed: yes
- Other examinations performed: clinical signs, body weight,organ weights, mortality

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Mortality:

No deaths

Clinical signs:

other: There were no signs of systemic reaction to treatment.

Gross pathology:

No macroscopic abnormalities were observed for the animals killed on Day 15.

Other findings:

Dermal reactions:
Transient slight to well defined dermal irritation (Grade 1 or 2 erythema and oedema) was evident in two males and two females on removal of the dressings, resolving completely by Day 5. In addition, desquamation (characterised by dryness/sloughing/scaling and/or spot/scab formation) was noted in two females during the first week of the study only. No dermal response to treatment was observed in the remaining six animals throughout the study.

Interpretation of results:

not classified

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

The acute lethal dermal dose to rats of this substance was demonstrated to be greater than 2000 mg/kg bodyweight.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

LD50

Value:

2 000 mg/kg bw

Quality of whole database:

1 (reliable without restriction)

Additional information

Acute oral toxicity:

An acute oral toxicity study (HLS, 1994) was conducted according to EU Method B1 which is key study. This study showed that the oral LD50 value of test substance is between 50 to 500 mg/kg body weight.

Acute dermal toxicity:

An acute dermal toxicity study (McRae, 1998) was conducted according to EU Method B3 which is key study. This study showed that the oral LD50 value of test substance is established to exceed 2000 mg/kg body weight.

Justification for selection of acute toxicity – oral endpoint
Study run to a method comparable with current guidelines and to GLP.

Justification for selection of acute toxicity – dermal endpoint
Study run to a method comparable with current guidelines and to GLP.

Justification for classification or non-classification

Oral LD50 = 50-500 mg/kg bw, Dermal LD50 >2000 mg/kg bw.

Therefore in accordance with Regulation (EC) No. 1272/2008 Table 3.1.1 the substance is classified as "Category 4" for acute oral toxicity and not classified for the acute dermal toxicity endpoint.