Benzonitrile, 4-[(1,6-dihydro-4-hydroxy-6-oxo-2-pyrimidinyl)amino]-

Endpoint summary

Description of key information

• Acute toxicity: Oral

Under the described conditions of the study performed according to OECD Guideline 423, the acute oral median lethal dose (LD50) of the test substance in the female outbred albino mouse was estimated to be greater than 2000 mg/kg bw (Sanders, 2004).

• Acute toxicity: Inhalation

No acute toxicity study via the inhalation route was available. However, this endpoint is waived as specific data are available for the oral and dermal exposure routes.

• Acute toxicity: Dermal

In an acute dermal toxicity study in male and female Crl:WI (Han) (SPF) rats, following the standard acute method according to OECD Guideline 402 and EC method B.3, the LD50 was established to exceed 2000 mg/kg body weight (Latour, 2016).

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:

2003-06-18 to 2003-07-15

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study without detailed documentation

Qualifier:

according to guideline

Guideline:

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

Deviations:

yes

Remarks:

Only a study summary was available for review which provided limited details on the test substance, tested animals, methodology and results.

GLP compliance:

no

Remarks:

The study was conducted in a facility operating to Good Laboratory Practice within the UK national GLP monitoring program, but the study report was not audited by the QA Unit. No formal claim of GLP compliance was made for this study.

Test type:

acute toxic class method

Limit test:

no

Species:

mouse

Strain:

other: outbred albino

Sex:

female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Weight at study initiation: 23-29 g
- Fasting period before study: animals were fasted before treatment but the fasting period was not provided

ENVIRONMENTAL CONDITIONS: no data

Route of administration:

oral: unspecified

Vehicle:

other: distilled water

Details on oral exposure:

VEHICLE
- Concentration in vehicle: no data
- Amount of vehicle (if gavage): no data
- Justification for choice of vehicle: no data
- Lot/batch no. (if required): no data
- Purity: no data


MAXIMUM DOSE VOLUME APPLIED:
- no data


DOSAGE PREPARATION:
- no data


CLASS METHOD (if applicable)
- Rationale for the selection of the starting dose: no data

The test item was administered orally as a suspension in distilled water.

Doses:

300 or 2000 mg/kg bw
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 of this dose level further groups of fasted females were treated at a dose level of 2000 mg/kg bodyweight. Dosing was performed sequentially.

No. of animals per sex per dose:

3 (1 group for 300 mg/kg bw and 2 groups for 2000 mg/kg bw)

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: All animals were examined for clinical signs and mortality at approximately 30 minutes, 1, 2, 3 and 4 hours after treatment and then daily during test days 1-14; body weights were recorded on day 0 (prior to administration) and on days 7 and 14. In addition, body weight gain was monitored during week 1 and week 2
- Necropsy of survivors performed: yes

Statistics:

no data

Preliminary study:

not applicable

Sex:

female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Mortality:

There were no deaths during the study.

Clinical signs:

other: There were no signs of systemic toxicity.

Gross pathology:

There were no abnormalities in macroscopic observations.

Other findings:

- Organ weights: no data
- Histopathology: no data
- Potential target organs: no data
- Other observations: no data

Interpretation of results:

GHS criteria not met

Conclusions:

Under the described conditions of the study, the acute oral median lethal dose (LD50) of the test substance in the female outbred albino mouse was estimated to be 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:

From 2015-12-22 to 2016-01-05

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

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.1200 (Acute Dermal Toxicity)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test type:

standard acute 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: I15CB0987
- Expiration date of the lot/batch: 01 March 2016 (retest date)
- Purity: 99.4%

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: at room temperature
- Solubility and stability of the test substance in the solvent/vehicle: stability for at least 6 hours at room temperature is confirmed over the concentration range 1 to 200 mg/mL.

TREATMENT OF TEST MATERIAL PRIOR TO TESTING
- Treatment of test material prior to testing:
The preparation (w/w) was kept at room temperature and dosed within 4 hours after adding the vehicle to the test item. Homogeneity was assessed by visual inspection of the solutions and the formulations were stirred during dosing, which ensures homogeneity sufficient for these kinds of studies. Adjustment was made for specific gravity of the vehicle. No correction was made for purity of the test item as the correction factor is 1.

Species:

rat

Strain:

Wistar

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: 5 male and 5 female rats (nulliparous and non-pregnant), Wistar strain Crl:WI (Han) (outbred, SPF-Quality); Charles River Deutschland, Sulzfeld, Germany
- Age at study initiation: Young adult animals (approx. 11 weeks old)
- Weight at study initiation: 200 - 332 grams
- Housing:group housed in Makrolon cages (MIV type, height 18 cm) during acclimatization period; Individually housed in labeled Makrolon cages (MIII type, height 18 cm.) containing sterilized sawdust as bedding material (Lignocel S 8-15, JRS - J.Rettenmaier & Söhne GmbH + CO. KG, Rosenberg, Germany) and paper as cage-enrichment (Enviro-dri, Wm. Lillico & Son (Wonham Mill Ltd), Surrey, United Kingdom)
- Diet (e.g. ad libitum): ad libitum, free access to pelleted rodent diet
- Water (e.g. ad libitum): ad libitum, free access to tap water
- Acclimation period: at least 5 days before start of treatment under laboratory conditions

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

IN-LIFE DATES: From 2015-12-22 to 2016-01-05

Type of coverage:

not specified

Vehicle:

propylene glycol

Details on dermal exposure:

TEST SITE
- Area of exposure: approx. 10% of the total body surface, i.e. approx. 25 cm² for males and 18 cm² for females
- % coverage: no data
- Type of wrap if used: 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.

REMOVAL OF TEST SUBSTANCE
- Washing (if done): after removal of dressing, the skin was cleaned of residual test item using tap water
- Time after start of exposure: 24 h

Duration of exposure:

24 h

Doses:

2000 mg/kg (single dosing)

No. of animals per sex per dose:

5

Control animals:

not specified

Details on study design:

Preparation of test item:
The preparation (w/w) was kept at room temperature and dosed within 4 hours after adding the vehicle to the test item. Homogeneity was assessed by visual inspection of the solutions and the formulations were stirred during dosing, which ensures homogeneity sufficient for these kinds of studies.
Adjustment was made for specific gravity of the vehicle. No correction was made for purity of the test item as the correction factor is 1.

Treatment of animals and application of test item:
Method: Dermal application. Test preparation was stirred on a magnetic stirrer during application.
Clipping: One day before exposure (Day -1) an area of approximately 5x7 cm on the back of each animal was clipped.

Frequency of dosing: Single dosage, on Day 1.

Observations:
Observation period: until day 15 after treatment
- Mortality/Viability: Twice daily.
- Body weights: Days 1 (pre-administration), 8 and 15.
- Clinical signs: At periodic intervals on the day of dosing (Day 1) and once daily thereafter, until Day 15. The time of onset, degree and duration were recorded and the symptoms graded according to fixed scales:
Maximum grade 4: grading slight (1) to very severe (4)
Maximum grade 3: grading slight (1) to severe (3)
Maximum grade 1: presence is scored (1).
- Necropsy At the end of the observation period, all animals were sacrificed by oxygen/carbon dioxide procedure and subjected to necropsy. Descriptions of all internal macroscopic abnormalities were recorded.

Statistics:

No statistical analysis was performed

Key result

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Mortality:

No mortality occurred.

Clinical signs:

other: Chromodacryorrhoea was noted for four male and two female animals on Day 1. Erythema maculate, scales, scabs were seen in the treated skin-area and/or right flank of five animals during the observation period. These local effects were considered not to ha

Gross pathology:

No abnormalities were found at macroscopic post mortem examination of the animals.

Interpretation of results:

GHS criteria not met

Conclusions:

The dermal LD50 value of JNJ-4370561-AAA (T002326) in Wistar rats was established to exceed 2000 mg/kg body weight. Based on these results, JNJ-4370561-AAA (T002326) does not have to be classified and has no obligatory labelling requirement for acute dermal toxicity according to the:
- Globally Harmonized System of Classification and Labelling of Chemicals (GHS) of the United Nations (2015) (including all amendments),
- Regulation (EC) No 1272/2008 on classification, labelling and packaging of substances and mixtures (including all amendments).

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

LD50

Value:

2 000 mg/kg bw

Additional information

Acute oral toxicity :

The study aimed to assess the acute oral toxicity of the substance to the mouse using the acute toxic class method (Sanders, 2004). Three groups of three females were administered orally with the test substance as a suspension in distilled water. A first group of three fasted females was treated with the test material at a dose level of 300 mg/kg bw. Based on the results from this dose level, another two groups of fasted females were treated at a dose level of 2000 mg/kg bw. Dosing was performed sequentially.

None of the animals died during the study. There were no signs of systemic toxicity, no abnormalities in body weight changes or macroscopic observations over the observation period of 14 days following administration of the test item. The acute lethal oral dose (LD50) of the substance to mice was > 2000 mg/kg bodyweight.

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 this substance a key study is available for the dermal route of exposure. Therefore, an acute inhalation toxicity study should not be performed.

Acute toxicity: dermal:

An acute dermal toxicity study with T002326 according to the standard acute method in male and female Crl:WI (Han) (SPF) rats (OECD guideline 402 and EU Method B.3 was performed (Latour, 2016). The substance was dissolved in propylene glycol and applied on a clipped area on the back at 2000 mg/kg bw. The 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. After 24 h hours of exposure remainings of the test item were washed-off. The animals were observed during 14 days. Lethality and viability were 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. Necropsy of survivors and macroscopic examination were also performed. Internal macroscopic abnormalities were recorded. No mortality occurred. Clinical signs observed were: Chromodacryorrhoea was noted for four male and two female animals on Day 1. Erythema maculate, scales, scabs were seen in the treated skin-area and/or right flank of five 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. No abnormalities were found at macroscopic post mortem examination of the animals.

The dermal LD50 value of T002326 in Wistar rats was established to exceed 2000 mg/kg body weight.

Justification for classification or non-classification

Based on the results showing an LD50 exceeding 2000 mg/kg bw, T002326 does not have to be classified and has no obligatory labelling requirement for acute oral toxicity according to the criteria laid down in Regulation (EC) No 1272/2008.

No data were available to decide on the classification for the inhalation route.

The dermal LD50 value of T002326 in Wistar rats was established to exceed 2000 mg/kg body weight. Based on these results, T002326 does not have to be classified and has no obligatory labelling requirement for acute dermal toxicity according to the criteria laid down in Regulation (EC) No 1272/2008.