4-[(6-chloro-2-[(4-cyanophenyl)amino]-4-pyrimidinyl]oxy]-3,5-dimethylbenzonitrile

Endpoint summary

Description of key information

 Acute toxicity: Oral:

In an K1 acute oral toxicity study in female Wistar rats, following the acute toxic class method in accordance with the OECD Guideline 423 and EU Method B.1 tris, the LD50 was established to be greater than 2000 mg/kg.

Acute toxicity: Inhalation:

In addition to the oral route of exposure, for substances other than gases, the information mentioned under REACH section 8.5.2 to 8.5.3 shall be provided for at least one other exposure route (REACH Regulation, column 2 adaptation of Annex VIII). For T002615, a key study is available for the dermal route of exposure. Therefore, an acute inhalation toxicity study should not be performed.

Acute toxicity: Dermal:

In an acute dermal toxicity study in male and female Wistar rats, following the standard acute method according to OECD Guideline 402 and EC method B.3, the LD50 was established to exceed 2500 mg/kg body weight (Arcelin, 2003).

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

Study period:

2003-01-21 to 2003-03-04

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method)

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method B.1 tris (Acute Oral Toxicity - Acute Toxic Class Method)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test type:

acute toxic class method

Limit test:

yes

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: 00403487
- Expiration date of the lot/batch: 2003-12-31
- Purity > 98%

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: in the container, at room temperature (range of 20 +/- 3 °C), away from direct light.
- Stability under test conditions: Stable for at least 2 hrs in PEG 300 at room temperature; determined at RCC Study No. 847009 (non-GLP study).
- Solubility and stability of the test substance in the solvent/vehicle: The vehicle was chosen after a non-GLP solubility trial which was performed before the study initation date.
- Reactivity of the test substance with the solvent/vehicle of the cell culture medium:

TREATMENT OF TEST MATERIAL PRIOR TO TESTING
The test item was weighed into a tared glass beaker on a suitable precision balance and the vehicle added (weight:volume).
Homogeneity of the test item in the vehicle was maintained during administration using a magnetic stirrer.

Species:

rat

Strain:

Wistar

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: RCC Ltd., Biotechnology and Animal Breeding Division CH-4414 Fullinsdorf/Switzerland
- Age at study initiation: Males: 8 weeks, Females: 12 weeks
- Weight at study initiation: Males: 242.2-259.3 g, Females: 186.7-192.5 g
- Fasting period before study: yes: for app. 18 h (access to water was permitted)
- Housing: In groups of three per sex in Makrolon type-4 cages with wire mesh tops and standard softwood bedding ('Lignocel' Schill AG, CH-4132 Muttenz/Switzerland).
- Diet: Pelleted standard Provimi Kliba 3433 rat/mouse maintenance diet, batch no. 67/02 (Provimi Kliba AG, CH-4303 Kaiseraugst/Switzerland) ad libitum.
- Water: community tap water from Fullinsdorf ad libitum
- Acclimation period: under laboratory conditions, after health examination (only animals without any visible signs of illness were used foor the study)

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 +/- 3 °C
- Humidity (%): 30-70 % (values above 70% during the cleaning process possible)
- Air changes (per hr): 10-15 air changes per hour
- Photoperiod (hrs dark / hrs light): 12 hrs fluorescent light/ 12 hrs dark (music during the light period)

Route of administration:

oral: gavage

Vehicle:

polyethylene glycol

Remarks:

PEG 300

Details on oral exposure:

VEHICLE
- Concentration in vehicle: 0.2 g/mL
- Justification for choice of vehicle: The vehicle was chosen after non-GLP solubility trial which was performed before the study initiation date.
- Lot/batch no. (if required): 435875/1 43002
- Purity: no data

MAXIMUM DOSE VOLUME APPLIED
-10 mL/kg bw

DOSAGE PREPARATION (if unusual)
- The dose formulations were made shortly before each dosing occasion using a magnetic stirrer and a spatula.
- The test substance was weighed into a tared glass beaker on a suitable balance and the vehicle added (Weight: volume).
- Homogeneity of the test substance in the vehicle was maintained during administration using a magnetic stirrer.

Doses:

2000 mg/kg bw

No. of animals per sex per dose:

3 females and 3 males per dose (2 groups at dose level of 2000 mg/kg bw)

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: Mortality/ Viability: daily during acclimatization and twice daily during days 1-15. Body weights: on test days 1 (prior to administration), 8 and 15. Clinical signs: daily during acclimatization and at app. 1, 2, 3 and 5 h after administration on test day 1 and once daily during days 2-15.
- Necropsy of survivors performed: yes

Statistics:

No statistical analysis was used.

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Mortality:

No deaths occured during the study.

Clinical signs:

other: Slightly ruffled fur and hunched posture was noted in all 2000 mg/kg treated female animals one hour after the treatment. No clinical signs were observed in the male animals during the course of the study.

Gross pathology:

No macroscopic findings were recorded at necroscopy.

Other findings:

no data

Interpretation of results:

GHS criteria not met

Conclusions:

The median lethal dose of T002615 after single oral administration to rats of both sexes observed over a period 14 days is: LD50 (rat) greater than 2000 mg/kg bw.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

LD50

Value:

2 000 mg/kg bw

Acute toxicity: via inhalation route

Link to relevant study records

Reference

Endpoint:

acute toxicity: inhalation

Data waiving:

other justification

Justification for data waiving:

other:

Endpoint conclusion

Endpoint conclusion:

no study available

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:

2003-01-21 to 2003-03-05

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 402 (Acute Dermal Toxicity)

Deviations:

no

Qualifier:

according to guideline

Guideline:

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

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test type:

fixed dose procedure

Limit test:

no

Specific details on test material used for the study:

- Name of test material (as cited in study report): T002615
- Substance type: no data
- Physical state: white solid
- Analytical purity: > 98 %
- Impurities (identity and concentrations): no data
- Composition of test material, percentage of components: no data
- Isomers composition: no data
- Purity test date: no data
- Lot/batch No.: 00403487
- Expiration date of the lot/batch: 2003-12-31
- Stability under test conditions: stable for at least 7 days in PEG 300 at room temperature; determined at RCC Study No. 846996 (Non GLP)
- Storage condition of test material: in the original container, at room temperature (range of 20 +/- 3 °C), away from direct sunlight
- Other: no data

Species:

rat

Strain:

Wistar

Remarks:

HanBrl: WIST (SPF)

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: RCC Ltd. Biotechnology & Animal Breeding Division, CH-4414 Füllinsdorf / Switzerland
- Age at study initiation: Males: 8-9 weeks; females: 11-12 weeks.
- Weight at study initiation: Male: 246.9-257.3 g; Females: 191.0-204.9 g.
- Fasting period before study: no data
- Housing: During acclimatization in groups of five per sex in Makrolon type-4 cages with standard softwood bedding. Individually in Makrolon type-3 cages with standard softwood bedding ("Lignocel", Schill AG, CH-4132 Muttenz) during treatment and observation
- Diet (e.g. ad libitum): Pelleted standard Provimi Kliba 3433, batch no. 67/02 rat maintenance diet (Provimi Kliba AG, CH-4303 Kaiseraugst) available ad libitum.
- Water (e.g. ad libitum): Community tap water from Itingen, available ad libitum.
- Acclimation period: 6 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 +/-3 °C
- Humidity (%): 30-70%
- Air changes (per hr): 10-15 air changes per hour
- Photoperiod (hrs dark / hrs light): 12 hrs flourescent light / 12 hrs dark

Type of coverage:

semiocclusive

Vehicle:

polyethylene glycol

Remarks:

PEG 300

Details on dermal exposure:

TEST SITE
- Area of exposure: back of the animals (free of fur)
- Type of wrap if used: dressing (the dressing was wrapped around abdomen and fixed with an elastic adhesive bandage)

REMOVAL OF TEST SUBSTANCE
- Washing (if done): yes (with lukewarm tap water and dried with disposable paper towels)
- Time after start of exposure: 24 hours after the application the dressing was removed

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 4 mL/kg bw
- Concentration (if solution): not applicable
- Constant volume or concentration used: yes
- For solids, paste formed: no

VEHICLE
- Amount(s) applied (volume or weight with unit): no data
- Concentration (if solution): a weight by volume dilution was prepared. Homogenity of the test item in the vehicle was maintained during administration using a magnetic stirrer.
- Lot/batch no. (if required):435875/1 43002
- Purity: no data

Duration of exposure:

24 h

Doses:

2000 mg/kg bw

No. of animals per sex per dose:

5 male and 5 female rats per dose

Control animals:

not required

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: mortaliy/viability: daily during acclimatization and twice daily during 1-15; body weights: on test days 1 ( prior to administration) , 8 and 15; clinical signs: daily during acclimatization and at app. 1, 2, 3 and 5 hours after administration on test day 1. Once daily during days 2-15.
- Necropsy of survivors performed: yes. All animals were killed at the end of the observation period by an intraperitoneal injection of Vetanarcol at a dose of at least 2.0 mL/kg/bw (equivalent to at least 324 mg sodium pentobarbitone/kg/bw) and discarded after macroscopic examinations were performed. No organs or tissues were retained.

Statistics:

No statistical analysis was used.

Preliminary study:

No data

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Mortality:

No deaths occurred during the study.

Clinical signs:

other: No systematic or local signs of toxicity were observed during the study period.

Gross pathology:

No macroscopic findings were observed at necroscopy.

Interpretation of results:

GHS criteria not met

Conclusions:

The median lethal dose of T002615 after single dermal administration to rats of both sexes observed over a period of 14 days is: LD50 (rat) greater than 2000 mg/kg bw.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

LD50

Value:

2 500 mg/kg bw

Additional information

Acute oral toxicity:

An acute oral toxicity study with T002615 according to the acute toxic class method in female Crl:WI (Han) (SPF) rats (OECD guideline 423 and EU Method B.1 tris) was performed (Arcelin, 2003). The test item was diluted in vehicle (PEG 300) at a concentration of 0.2 g/ml and administered at a volume dosage of 10 ml/kg. The animals were examined daily during the acclimatization period and mortality, viability and clinical signs were recorded. All animals were examined for clinical signs at approximately 1, 2, 3 and 5 hours after treatment on day 1 and once daily during test days 2-15. Mortality/viability was recorded twice daily during test days 1-15. Body weights were recorded on day 1 (prior to administration) and on days 8 and 15. All animals were necropsied and examinedmacroscopically. All animals survived until the end of the study period. Slightly ruffled fur and hunched posture was noted in all 2000 mg/kg treated female animals one hour after the treatment. No clinical signs were observed in the male animals during the course of the study. The body weight of the animals was within the range commonly recorded for this strain and age. No macroscopic findings were recorded at necropsy. The LD50 was established to be greater than 2000 mg/kg. This study was used as key study.

In addition, a supporting acute oral toxicity study was available which assessed the acute oral toxicity following a single oral administration in female outbred albino mouse.

A group of three fasted females was treated with the test material at a dose level of 300 mg/kg bodyweight. Based on the results from this dose level further groups of fasted females were treated at a dose level of 2000 mg/kg bodyweight. Dosing was performed sequentially. The test item was administered orally as a suspension in arachis oil BP. Clinical signs and bodyweight development were monitored during the study. All animals were subjected to gross necropsy.

The acute oral median lethal dose (LD50) of the test item in the female outbred albino mouse was estimated as being greater than 2500 mg/kg bodyweight.

As much less details were available on test item and environmental conditions and as the results of the study were similar as in the key study, this K2 study was used as supporting evidence.

 

Acute inhalation toxicity:

In addition to the oral route of exposure, for substances other than gases, the information mentioned under REACH section 8.5.2 to 8.5.3 shall be provided for at least one other exposure route (REACH Regulation, column 2 adaptation of Annex VIII). For this test item a key study is available for the dermal route of exposure. Therefore, an acute inhalation toxicity study should not be performed.

 

Acute dermal Toxicity:

An acute dermal toxicity study with T002615 according to OECD guideline 402 and EU Method B.3 in male and female Crl:WI (Han) (SPF) rats was performed (Arcelin G., 2003). The test item was dissolved in 1% aqueous CMC (carboxymethyl cellulose) and applied on a clipped area on the back at 2000 mg/ kg bw. The test item formulation was held in contact with the skin with a dressing consisting of a surgical gauze patch (Surgy 1D), successively covered with aluminum foil and Coban elastic bandage. A piece of Micropore tape was additionally used for fixation of the bandages in females only. After 24 hours of exposure remainings of the test item were washed-off using tap water. The animals were observed during 14 days. Mortality/viability was observed twice daily, and bodyweight was monitored at day 1 (prior to administration), 8 and 15. Clinical signs were observed at periodic intervals on the day of dosing (day 1) and once daily thereafter, until day 15. Necropsy of survivors and macroscopic examination were also performed. Internal macroscopic abnormalities were recorded.

No mortality occurred. Clinical signs observed were: Chromodacryorrhoea (nose) was noted for two male and three female animals on Days 1 and/or 2. General erythema, scars, scales, scabs and/or white staining were seen in the treated skin-area of all animals during the observation period. These local effects were considered not to have affected the conclusion of the study.

The changes noted in body weight gain in males and females were within the range expected for rats used in this type of study and were therefore considered not indicative of toxicity.

Abnormalities of the epididymides (right side, tail: yellowish, soft nodule) or liver (left lateral lobe: irregular surface) were found in two male animals at macroscopic post mortem examination. Macroscopic post mortem examination of the other animals did not reveal any abnormalities.

The dermal LD50 value of T002615 in Wistar rats was established to exceed 2000 mg/kg body weight.

Justification for classification or non-classification

Acute oral toxicity:

Based on the available data and the criteria of the CLP Regulation, this test item should not be classified for acute oral toxicity.

Acute inhalation toxicity:

No data were available for the inhalation route so no classification can be derived.

Acute dermal toxicity:

Based on the available data and the criteria of the CLP Regulation, this test item should not be classified for acute dermal toxicity.