6,10-Dioxaspiro[4.5]decane-7-acetic acid, 9-[2-[2-(4-fluorophenyl)-5-(1-methylethyl)-3-phenyl-4-[(phenylamino)carbonyl]-1H-pyrrol-1-yl]ethyl]-, 1,1-dimethylethyl ester, (7R,9R)-

Eye irritation

Administrative data

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

From 02 to 09 February 2009

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

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

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2009

Report date:

2009

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Deviations:

no

GLP compliance:

yes

Test material

Test material information

Constituent 1

Reference substance name:

tert-butyl 2-[(7R,9R)-9-{2-[2-(4-fluorophenyl)-3-phenyl-4-(phenylcarbamoyl)-5-(propan-2-yl)-1H-pyrrol-1-yl]ethyl}-6,10-dioxaspiro[4.5]decan-7-yl]acetate

EC Number:

920-205-7

Cas Number:

1035204-85-3

Molecular formula:

C42H49FN2O5

IUPAC Name:

tert-butyl 2-[(7R,9R)-9-{2-[2-(4-fluorophenyl)-3-phenyl-4-(phenylcarbamoyl)-5-(propan-2-yl)-1H-pyrrol-1-yl]ethyl}-6,10-dioxaspiro[4.5]decan-7-yl]acetate

Details on test material:

Batch No.: TCA-153-74
Purity: 98%

Test animals / tissue source

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Source: Harlan France SARL, Gannat, France
- Age at study initiation: 10-20 weeks old
- Weight at study initiation: 2-4 kg
- Housing: Animals were individually housed in labeled cages with perforated floors and shelters.
- Diet (e.g. ad libitum): Pelleted diet for rabbits was approximately 100 grams per day. Hay was provided at least three times a week.
- Water (e.g. ad libitum): Free access to tap water
- Acclimation period: At least 5 days before start of treatment under laboratory conditions.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21.0 ± 3.0 ℃ (actual range: 21.1-22.7 ℃)
- Humidity (%): 30-70% (actual range: 42-65%)
- Air changes (per hr): Approximately 15 air changes per hour
- Photoperiod (hrs dark / hrs light): 12 hours artificial fluorescent light and 12 hours darkness per day

No additional data

Test system

Vehicle:

unchanged (no vehicle)

Controls:

not required

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): on average 37.0 mg (range 36.9-37.0 mg) of the test substance (a volume of approximately 0.1 mL)

No additional data

Duration of treatment / exposure:

72 h

Observation period (in vivo):

72 h

Number of animals or in vitro replicates:

3 males

Details on study design:

The study was performed in a stepwise manner and was started by treatment of a single rabbit (sentinel). The two other animals were treated in a similar manner 1 week later, after considering the degree of eye irritation observed in the first animal.

Animals were treated by instillation of the test substance 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 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.

After the final observation, the animals were sacrificed by intra-venous injection of Euthasol® 20%.

Results and discussion

In vivo

Results

Results 1

Irritation parameter:

cornea opacity score

Basis:

mean

Time point:

other: Mean of 24, 48 and 72 hours

Score:

0

Max. score:

0

Results 2

Irritation parameter:

iris score

Basis:

mean

Time point:

other: mean of 24, 48 and 72 hours

Score:

0

Max. score:

0

Results 3

Irritation parameter:

conjunctivae score

Basis:

mean

Time point:

other: Mean of 24, 48 and 72 hours

Score:

0.3

Max. score:

1

Reversibility:

fully reversible within: 48 hours

Remarks on result:

other: Redness were observed at 1 hour and 24 hour in all of three animals.

Results 4

Irritation parameter:

chemosis score

Basis:

mean

Time point:

other: Mean of 24, 48 and 72 hours

Score:

0

Max. score:

1

Reversibility:

fully reversible within: 24 hours

Remarks on result:

other: Chemosis were observed at 1 hour in all of three animals.

Irritant / corrosive response data:

Irritation
Instillation of approximately 37 mg of the test substance into one eye of each of three rabbits resulted in irritation of the conjunctivae, which consisted of redness, chemosis and discharge. The irritation completely resolved within 48 hours.
No iridial irritation or corneal opacity was observed, and treatment of the eyes with 2% fluorescein 24 hours after test substance instillation revealed no corneal epithelial damage.

Corrosion
There was no evidence of ocular corrosion.

Other effects:

Coloration/Remnants
No staining of (peri) ocular tissues by the test substance was observed. Remnants of the test substance were present in the eye on day 1.

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

Any other information on results incl. tables

Individual eye irritation scores

Animal	Time after dosing	Cornea			Iris	Conjunctivae
		Opacity (0-4)	Area (0-4)	Fluor area (%)*	(0-2)	Redness (0-3)	Chemosis (0-4)	Discharge (0-3)
121#	1 h	0	0	0	0	1	1	1
	24 h	0	0		0	1	0	0
	48 h	0	0		0	0	0	0
	72 h	0	0		0	0	0	0
116	1 h	0	0	0	0	1	1	1
	24 h	0	0		0	1	0	0
	48 h	0	0		0	0	0	0
	72 h	0	0		0	0	0	0
128	1 h	0	0	0	0	1	1	1
	24 h	0	0		0	1	0	0
	48 h	0	0		0	0	0	0
	72 h	0	0		0	0	0	0

# Sentinel

* Green staining after fluorescein treatment (percentage of total corneal area)

Applicant's summary and conclusion

Interpretation of results:

not irritating

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

Treatment with the test substance caused reversible effects on the conjunctivae of three (3/3) rabbits.