4-[(4-HYDROXYPYRIMIDIN-2-YL)AMINO]BENZONITRILE

Eye irritation

Administrative data

Endpoint:

eye irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

FRom 2004-11-01 to 2004-11-01

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

study well documented, meets generally accepted scientific principles, acceptable for assessment

Remarks:

While only a study summary was available for review which provided limited details on the test substance and methodology, sufficient information was provided to deem the study reliable with restrictions.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2004

Report date:

2004

Materials and methods

Test guideline

Qualifier:

no guideline followed

Principles of method if other than guideline:

The ocular irritancy potential of the test material was assessed using the rabbit enucleated eye test (REET) SPL Standard Test Method 569.04. This method involved the application of the test material onto the cornea of the enucleated eye.

GLP compliance:

no

Remarks:

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

Test material

Test material information

Constituent 1

Reference substance name:

4-[(4-Oxo-1,4-dihydropyrimidin-2-yl)amino]benzonitrile

EC Number:

606-195-4

Cas Number:

189956-45-4

Molecular formula:

C11H8N4O

IUPAC Name:

4-[(4-Oxo-1,4-dihydropyrimidin-2-yl)amino]benzonitrile

Test material form:

solid: particulate/powder

Details on test material:

- Name of test material (as cited in study report): JNJ-40370226-AAA (T002487)
- Physical state: solid (powder)
- Appearance: Off-white powder

Specific details on test material used for the study:

no data

Test animals / tissue source

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

SOURCE OF COLLECTED EYES
- Source: New Zealand White rabbits, no further data on test animals
- Storage, temperature and transport conditions of ocular tissue (e.g. transport time, transport media and temperature, and other conditions): the enucleated eyes were maintained at a temperature of 32°C +/- 1.5°C within the superfusion apparatus.

Test system

Vehicle:

not specified

Controls:

yes, concurrent negative control

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.1 mL of the test item, which had been found to weigh approximately 60 mg
- Concentration (if solution): no data

NEGATIVE CONTROL
- Amount(s) applied (volume or weight with unit): No data
- Concentration (if solution): 0.9%

Duration of treatment / exposure:

single application

Number of animals or in vitro replicates:

3 enucleated eyes were treated with the test item and 2 enucleated eyes were treated with the control.

Details on study design:

REMOVAL OF TEST SUBSTANCE: no data

OBSERVATION TIME POINTS
- corneal opacity and corneal epithelium condition: 60, 120, 180 and 240 min post dosing
- fluorescein uptake: 240 min post dosing
- corneal swelling: 60, 120 and 240 min post dosing

SCORING SYSTEM:
The direct effect of the test substance on the cornea was assessed by evaluation of corneal thickness, corneal opacity, alteration of corneal epithelium and fluorescein uptake, throughout the duration of the study.


TOOL USED TO ASSESS SCORE:
no data

Results and discussion

In vitro

Results

Results 1

Irritation parameter:

cornea opacity score

Remarks:

cloudiness/ mean of 3 eyes after 60, 120, 180 and 240 min post dosing

Run / experiment:

1

Value:

1

Vehicle controls validity:

not applicable

Negative controls validity:

valid

Positive controls validity:

not applicable

Remarks on result:

positive indication of irritation

Results 2

Irritation parameter:

cornea opacity score

Remarks:

area/mean of three eyes after 60, 120, 180 and 240 min post dosing

Run / experiment:

1

Value:

2.3

Vehicle controls validity:

not applicable

Negative controls validity:

valid

Positive controls validity:

not applicable

Results 3

Irritation parameter:

other: Fluorescein uptake

Remarks:

intensity/mean of three eyes

Run / experiment:

1

Value:

1

Vehicle controls validity:

not applicable

Negative controls validity:

valid

Positive controls validity:

not applicable

Remarks on result:

positive indication of irritation

Results 4

Irritation parameter:

other: Fluorescein uptake

Remarks:

area/mean of three eyes

Run / experiment:

1

Value:

2

Vehicle controls validity:

not applicable

Negative controls validity:

valid

Positive controls validity:

not applicable

Results 5

Irritation parameter:

percent corneal swelling

Remarks:

mean of three eyes 60 min post dosing

Run / experiment:

1

Value:

13.7

Vehicle controls validity:

not applicable

Negative controls validity:

valid

Positive controls validity:

not applicable

Remarks on result:

positive indication of irritation

Results 6

Irritation parameter:

percent corneal swelling

Remarks:

mean of three eyes 120 min post dosing

Run / experiment:

1

Value:

15.3

Vehicle controls validity:

not applicable

Negative controls validity:

valid

Positive controls validity:

not applicable

Remarks on result:

positive indication of irritation

Results 7

Irritation parameter:

percent corneal swelling

Remarks:

mean of three eyes 240 min post dosing

Run / experiment:

1

Value:

19.6

Vehicle controls validity:

not applicable

Negative controls validity:

valid

Positive controls validity:

not applicable

Remarks on result:

positive indication of irritation

Other effects / acceptance of results:

negative control eyes:
- corneal opacity: 0 (mean of 2 eyes)
- corneal epithelium condition: normal (2 eyes)
- fluorescein uptake: 0 (mean of 2 eyes)
- corneal swelling: 1.0 at 60 min post dosing, 1.1 at 120 minutes post dosing and 1.3 at 240 minutes post dosing

Corneal epithelium condition was normal at all timepoints for all test item treated eyes.

Any other information on results incl. tables

Table 1. Results obtained for corneal opacity at 60, 120, 180, and 240 minutes post exposure.

	Corneal Opacity
	Observation Period (minutes post dosing)
	60		120		180		240
	Cldy	Area	Cldy	Area	Cldy	Area	Cldy	Area
Test Eyes	1	3	1	4	1	4	1	4
	1	3	1	2	1	1	1	1
	1	2	1	2	1	1	1	1
Control Eyes	0	0	0	0	0	0	0	0
	0	0	0	0	0	0	0	0

Cldy = Corneal opacity

 

 

Table 2. Results obtained for corneal epithelium condition at 60, 120, 180, and 240 minutes post exposure.

	Corneal Epithelium Condition
	Observation Period (minutes post dosing)
	60	120	180	240
Test Eyes	Normal	Normal	Normal	Normal
	Normal	Normal	Normal	Normal
	Normal	Normal	Normal	Normal
Control Eyes	Normal	Normal	Normal	Normal
	Normal	Normal	Normal	Normal

 

 

Table 3. Results obtained for fluorescein uptake 240 minutes post exposure.

	Fluorescein Uptake (240 minutes)
	Test Eyes			Control Eyes
Int	1	1	1	0	0
Area	4	1	1	0	0

Int = Intensity of fluorscein uptake

 

Table 4. Results obtained for corneal swelling at 60, 120, and 240 minutes post exposure.

	Corneal Swelling (%)(minutes post dosing)
	60	120	240
Test Eyes	13.7	15.3	19.6
Control Eyes	1.0	1.1	1.3

Test eyes results are the mean of three eyes while control eyes are the mean of two eyes

Applicant's summary and conclusion

Interpretation of results:

Category 1 (irreversible effects on the eye) based on GHS criteria

Conclusions:

The test material is considered to have the potential to cause severe ocular irritancy in vivo.