2-Propen-1-one, 3-(dimethylamino)-1-(3-pyridinyl)-

Eye irritation

Administrative data

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

13 June - 04 July 2000

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: The study has been performed according to OECD and/or EC guidelines and according to GLP principles.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2000

Report date:

2000

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method B.5 (Acute Toxicity: Eye Irritation / Corrosion)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

STI571 Y5A

IUPAC Name:

STI571 Y5A

Details on test material:

- Name of test material (as cited in study report): STI571 Y5A
- Substance type: Brown solid
- Physical state: solid
- Analytical purity: >95%
- Lot/batch No.: 0093800007
- Expiration date of the lot/batch: 08 May 2001 (allocated by NOTOX, 1 year after receipt of the test substance)
- Storage condition of test material: In refrigerator in the dark.
- Purity: Not indicated by sponsor; treated as 100% pure
- Stability under storage conditions: Not indicated
- Stability in vehicle: see details on Analytical verification of doses or concentrations.

Test animals / tissue source

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Source: Charles River Deutschland, Kisslegg, Germany.
- Age at study initiation: Animals used within the study were at least 6 weeks old*
- Weight at study initiation: less than 3.5 kg*
- Housing: Individually housed in labelled cages with perforated floors (Scanbur, Denmark) and equipped with an automatic drinking system (ITL, Bergen, The Netherlands).
- Diet (e.g. ad libitum): Standard laboratory rabbit diet (Charles River Breeding and Maintenance Diet for Rabbits, Altromin, Germany) approx. 100 g. per day. In addition, hay (BMI, Helmond, the Netherlands) was provided once a week.
- Water (e.g. ad libitum): Free access to tap water.
- Acclimatization period: at least 5 days before start of treatment under laboratory conditions.

* Animal specifications (sex, age and body weight) are specified in the attached table.

A health inspection was performed prior to commencement of treatment, to ensure that the animals were in a good state of health. Special attention was paid to the eyes, which were free from any abnormality.

Results of analysis for diet (nutrients and contaminants), hay and water were assessed and did not reveal any findings that were considered to have affected the study integrity. All certificates and results of analysis are retained in the NOTOX archives.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21ºC
- Humidity (%): 50%
- Air changes (per hr): 15
- Photoperiod (hrs dark / hrs light): 12 hours artificial fluorescent light and 12 hours darkness per day
Deviations from these optimal conditions were noted, but were considered not to have affected study integrity.

IN LIFE DATES: From: 13 June to 04 July 2000

Test system

Vehicle:

unchanged (no vehicle)

Controls:

other: One eye of each animal remained untreated and served as the reference control.

Amount / concentration applied:

TEST MATERIAL- Amount(s) applied (volume or weight with unit):
72.4 ± 2.6 mg (a volume of approximately 0.1 ml)

Duration of treatment / exposure:

Single instillation on Day 1.

Observation period (in vivo):

The eyes of each animal were examined approximately 1, 24, 48 and 72 hours and 7, 14 and 21 days after instillation of the test substance

Number of animals or in vitro replicates:

3

Details on study design:

STUDY DESIGN
Each animal was treated by instillation of 72.4 ± 2.6 mg of the test substance (a volume of approximately 0.1 ml) in the conjunctival sac of one of the eyes after gently pulling the lower lid away from the eyeball. The lids were then gently held together for about one second to prevent loss of the test substance. The other eye remained untreated and served as the reference control.

Immediately after the 24 hour observation, a solution of 2% fluorescein in water (adjusted to pH 7.0) was instilled into both eyes of each animal to quantitatively determine corneal epithelial damage. When considered necessary, this procedure was repeated to assess recovery. Any bright green stained area, indicating epithelial damage, was estimated as a percentage of the total corneal area.

OBSERVATIONS
- Mortality/Viability: Twice daily.
- Toxicity: At least once daily.
- Body Weight: Day of treatment (prior to instillation).
- Irritation:
The eyes of each animal were examined approximately 1, 24, 48 and 72 hours and 7, 14 and 21 days after instillation of the test substance.
The irritation scores and a description of all other (local) effects were recorded.The irritation was assessed according to the following numerical scoring system. At each observation, the highest scores given were recorded:

CORNEAL IRRITATION
Opacity: degree of density (area most dense taken for reading)
0: No ulceration or opacity (may include slight dulling of normal luster)
1: Scattered or diffuse areas of opacity, details of iris clearly visible
2: Easily discernible translucent area, details of iris slightly obscured
3: Nacreous area, no details of iris visible, size of pupil barely discernible
4: Opaque cornea, iris not discernible through the opacity

Area of cornea involved:
0: No ulceration or opacity
1: One quarter or less but not zero
2: Greater than one quarter, but less than half
3: Greater than half, but less than three quarters
4: Greater than three quarters, up to whole area

IRIS
0: Normal
1: Markedly deepened rugae, congestion, swelling, moderate circumcorneal hyperaemia, or injection, any of these or combination thereof, iris still reacting to light (sluggish reaction is positive)
2: No reaction to light, hemorrhage, gross destruction (any or all of these)

CONJUNCTIVAL IRRITATION
Redness (refers to palpebrae and sclera, excluding cornea and iris):
0: Blood vessels normal
1: Some blood vessels definitely hyperaemic (injected)
2: Diffuse, crimson color, individual vessels not easily discernible
3: Diffuse beefy red

Chemosis (refers to lids and/or nictitating membranes):
0: No swelling
1: Any swelling above normal (includes nictitating membranes)
2: Obvious swelling with partial eversion of lids
3: Swelling with lids about half closed
4: Swelling with lids more than half closed

Discharge:
0: No discharge (may include small amounts observed in inner canthus of normal animals)
1: Any amount different from normal and/or lacrimation
2: Discharge with moistening of the lids and hairs just adjacent to lids
3: Discharge with moistening of the lids and hairs (considerable area around the eye)

Where standard lighting was considered inadequate for observing minor effects, eye examinations were performed using an ophthalmic examination lamp.
In cases of equivocal results when comparing the treated and untreated eyes, the illustrated guide from the Consumer Product Safety Commission, Washington, D.C. 20207 was used for additional control purposes.

Results and discussion

In vivo

Results

Results 1

Irritation parameter:

cornea opacity score

Remarks:

(opacity)

Basis:

animal #1

Remarks:

(mean)

Time point:

other: 24, 48 and 72 hrs.

Score:

0.3

Max. score:

4

Reversibility:

fully reversible within: 48 hours

Results 2

Irritation parameter:

cornea opacity score

Remarks:

(opacity)

Basis:

animal #2

Remarks:

(mean)

Time point:

other: 24, 48 and 72 hrs.

Score:

0.7

Max. score:

4

Reversibility:

fully reversible within: 72 hours

Results 3

Irritation parameter:

cornea opacity score

Remarks:

(opacity)

Basis:

animal #3

Remarks:

(mean)

Time point:

other: 24, 48 and 72 hrs.

Score:

1

Max. score:

4

Reversibility:

fully reversible within: 14 days

Remarks on result:

other: pannus (neovascularisation of the cornea) noted at 7 days after instillation.

Results 4

Irritation parameter:

iris score

Basis:

animal #1

Remarks:

(mean)

Time point:

other: 24, 48 and 72 hrs.

Score:

0

Max. score:

2

Results 5

Irritation parameter:

iris score

Basis:

animal #2

Remarks:

(mean)

Time point:

other: 24, 48 and 72 hrs.

Score:

0

Max. score:

2

Results 6

Irritation parameter:

iris score

Basis:

animal #3

Remarks:

(mean)

Time point:

other: 24, 48 and 72 hrs.

Score:

0

Max. score:

2

Results 7

Irritation parameter:

conjunctivae score

Remarks:

(redness)

Basis:

animal #1

Remarks:

(mean)

Time point:

other: 24, 48 and 72 hrs.

Score:

2.3

Max. score:

3

Reversibility:

fully reversible within: 14 days

Results 8

Irritation parameter:

conjunctivae score

Remarks:

(redness)

Basis:

animal #2

Remarks:

(mean)

Time point:

other: 24, 48 and 72 hrs.

Score:

2.3

Max. score:

3

Reversibility:

fully reversible within: 14 days

Results 9

Irritation parameter:

conjunctivae score

Remarks:

(redness)

Basis:

animal #3

Remarks:

(mean)

Time point:

other: 24, 48 and 72 hrs.

Score:

2.3

Max. score:

3

Reversibility:

fully reversible within: 14 days

Remarks on result:

other: reduced elasticity of the eyelids observed at 48 hours, and 7 and 14 days following instillation.

Results 10

Irritation parameter:

chemosis score

Basis:

animal #1

Remarks:

(mean)

Time point:

other: 24, 48 and 72 hrs.

Score:

1

Max. score:

4

Reversibility:

fully reversible within: 72 hours

Results 11

Irritation parameter:

chemosis score

Basis:

animal #2

Remarks:

(mean)

Time point:

other: 24, 48 and 72 hrs.

Score:

1.3

Max. score:

4

Reversibility:

fully reversible within: 7 days

Results 12

Irritation parameter:

chemosis score

Basis:

animal #3

Remarks:

(mean)

Time point:

other: 24, 48 and 72 hrs.

Score:

1.7

Max. score:

4

Reversibility:

fully reversible within: 14 days

Irritant / corrosive response data:

Irritation
Instillation of approximately 72 mg of the test substance (a volume of approximately 0.1 ml) into one eye of each of three rabbits resulted in effects on the cornea, iris and conjunctivae. Corneal injury was seen as opacity (maximum grade 1) and epithelial damage (maximum 25% of the corneal area). As a result of the corneal injury, pannus (neovascularisation of the cornea) was apparent in one animal 7 days after instillation. The corneal injury had resolved within 72 hours in two animals and within 14 days in the other animal. Iridial irritation (grade 1) was observed in two animals after 1 hour. Reduced elasticity of the eyelids was noted in one animal after 48 hours and after 7 and 14 days. Irritation of the conjunctivae was seen as redness, chemosis and discharge, which had completely resolved within 14 days in two animals and within 21 days in the other animal.

Corrosion
There was no evidence of ocular corrosion.

Individual and mean eye irritation scores are specified in the attached table.

Any other information on results incl. tables

Colouration

Yellow staining of the fur on the head and paws, caused by the test substance, was noted on all animals throughout the observation period.

 

Toxicity / Mortality

No symptoms of systemic toxicity were observed in the animals during the test period and no mortality occurred.

Applicant's summary and conclusion

Conclusions:

Based on these results and according to the EC criteria for classification and labelling requirements for dangerous substances and preparations (Guidelines in Commission Directive 93/21/EEC), STI571 Y5A does not have to be classified and has no obligatory labelling requirement for eye irritation.