2-{2-[(2R,4R,6R)-4,6-dihydroxytetrahydro-2H-pyran-2-yl]ethyl}-1H-isoindole-1,3(2H)-dione

Eye irritation

Administrative data

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

28 January 2008 - 07 February 2008

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Study performed in accordance with OECD and EC guidelines and conducted to GLP.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2008

Report date:

2008

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

Test guideline 3

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.2400 (Acute Eye Irritation)

Deviations:

no

Test guideline 4

Qualifier:

according to guideline

Guideline:

other: Japanese Ministry of Agriculture, Forestry and Fisheries (JMAFF), 12 Nousan, Notification No. 8147, November 2000, including the most recent partial revisions.

Deviations:

no

GLP compliance:

yes

Test material

Test material information

Constituent 1

Reference substance name:

2-{2-[(2R,4R,6R)-4,6-dihydroxyoxan-2-yl]ethyl}-2,3-dihydro-1H-isoindole-1,3-dione

EC Number:

700-188-0

Cas Number:

1333481-90-5

Molecular formula:

C15H17NO5

IUPAC Name:

2-{2-[(2R,4R,6R)-4,6-dihydroxyoxan-2-yl]ethyl}-2,3-dihydro-1H-isoindole-1,3-dione

Constituent 2

Reference substance name:

Lactol

IUPAC Name:

Lactol

Constituent 3

Reference substance name:

Phthalamido Lactol

IUPAC Name:

Phthalamido Lactol

Details on test material:

- Name of test material (as cited in study report): Lactol
- Physical state: Solid (White powder)
- Analytical purity: Not indicated by the sponsor; treated as 100% Pure
- Impurities (identity and concentrations): Not indicated
- Lot/batch No.: PDC-010-194
- Expiration date of the lot/batch: 13 December 2008 (allocated by NOTOX, 1 year after receipt of the test substance)
- Stability under test conditions: Stable
- Storage condition of test material: At room temperature in the dark.

Test animals / tissue source

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Source: Harlan, Horst, The Netherlands
- Age at study initiation: At least 10 weeks old.
- Weight at study initiation: At least 2.0 kg.
- Housing:Individually in cages with perforated floors (Scanbur, Denmark, dimensions 56 x 44 x 37.5 cm).
- Diet (e.g. ad libitum): Pelleted diet for rabbits (K-H from SSNIFF® Spezialdiaten GmbH, Soest, Germany) approximately 100 grams per day. Hay (TechniLab-BMI BV, Someren, The Netherlands) 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 the start of treatment under laboratory conditions.


ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21.0 ± 3.0°C (actual range: 18.3 - 21.3°C)
- Humidity (%): 30-70% (actual range 41-93%)
- Air changes (per hr): 15
- Photoperiod (hrs dark / hrs light): 12 hours artificial fluorescent light and 12 hours darkness per day.


IN-LIFE DATES: From: 28 January 2008 To: 07 February 2008

Test system

Vehicle:

unchanged (no vehicle)

Controls:

no

Amount / concentration applied:

Average 31 mg (Range 30.8 - 31.4 mg) of the test substance (a volume of approximately 0.1 mL).

Duration of treatment / exposure:

Single exposure

Observation period (in vivo):

7 days (observations made at 1, 24, 48, and 72 hours, and 7 days after exposure).

Number of animals or in vitro replicates:

3 males.

Details on study design:

REMOVAL OF TEST SUBSTANCE
- Washing (if done): N/A
- Time after start of exposure:


SCORING SYSTEM:

The eyes of each animal were examined approximately 1, 24, 48, 72 hours and/or 7 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)
No ulceration or opacity (may include slight dulling of normal lustre) 0
Scattered or diffuse areas of opacity, details of iris clearly visible 1
Easily discernible translucent area, details of iris slightly obscured 2
Nacreous area, no details of iris Visible, size of pupil barely discernible 3
Opaque cornea, iris not discernible through the opacity . 4

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

IRIS
Normal 0
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) 1
No reaction to light, hemorrhage, gross destruction (any or all of these) 2

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

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

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

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.


TOOL USED TO ASSESS SCORE: Immediatelt after the 24-hour abservation, a solution of 2% Fluorescin in water was instilled into both eyes of each animal to quantitatively determie corneal damage.

Results and discussion

In vivo

Results

Irritation parameter:

overall irritation score

Basis:

mean

Time point:

other: 24, 48 and 72 hours

Score:

>= 0.3 - <= 1.3

Max. score:

2

Reversibility:

fully reversible within: 7 days

Irritant / corrosive response data:

Refer to table 1, below and figure 1, attached.
Installation of approximately 31 mg of LACTOL (a volume of approximately 0.1 mL) into one eye of each of three rabbits resulted in irritation of the
conjuntivae, which consisted of redness, chemosis and discharge. The irritation had completely resolved within 7 days in one animal and within 72
hours in the other two animals. No iridial irritation or corneal opacity were observed, and treatment of the eyes with 2% flourescein, 24 hours after
test substance installation revealedno corneal epithelial damage.

Other effects:

Corrosion
There was no evidence of ocular corrosion.

Colouration I Remnants
No staining of (peri) ocular tissues by the test substance was observed and no test substance remnants were seen.

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

Any other information on results incl. tables

Table 1: Individual eye irritation scores.

Animal	Time after dosing	Cornea			Iris	Conjunctivae			Comments
		Opacity (0-4)	Area (0-4)	Fluor Area (%)^2	(0-2)	Redness (0-3)	Chemosis (0-4)	Discharge (0-3)
710^1	1 hour	0	0	-	1	2	1	1
	24 hours	0	0	0	0	2	0	0
	48 hours	0	0	-	0	1	0	0
	72 hours	0	0	-	0	1	0	0
	7 days	0	0	-	0	0	0	0
714	1 hour	0	0	-	0	2	1	1
	24 hours	0	0	0	0	1	0	0
	48 hours	0	0	-	0	0	0	0
	72 hours	0	0	-	0	0	0	0
716	1 hour	0	0	-	0	2	1	1
	24 hours	0	0	0	0	2	0	0
	48 hours	0	0	-	0	1	0	0
	72 hours	0	0	-	0	0	0	0

¹Sentinel, ²Green staining after fluorescein treatment (percentage of corneal area).

Applicant's summary and conclusion

Interpretation of results:

not irritating

Remarks:

Migrated information Criteria used for interpretation of results: other: EU and GHS

Conclusions:

Treatment with Lactol caused reversible effects on the cornea, iris and conjunctivae, in the eye of three (3/3) rabbits. However, it can be classified as non irritating according to the EU criteria.

Executive summary:

An eye irritation study was performed by NOTOX B.V., The Netherlands on behalf of Pfizer Ireland Pharmaceuticals, Ltd to assess the potential for eye irritation of the test substance, Lactol. The test was performed under GLP according to OECD guideline 405 and EC guideline B5, and also to US and Japanese guidelines.

Approximately 31 mg of test substance were applied to the conjunctival sac of one of the eyes of each of three male rabbits, and observations taken after 1, 24, 48, and 72 hours, and after 7 days.

Instillation of the test substance resulted in irritation of the conjunctivae, which consisted of redness, chemosis and discharge. The irritation had completely resolved within 7 days in one animal and within 72 hours in the other two animals. No iridial irritation or corneal opacity were observed, and treatment of the eyes with 2% fluorescein, 24 hours after test substance instillation, revealed no corneal epithelial damage. Treatment with LACTOL caused reversible effects on the cornea, iris and conjunctivea in the eye of three (3/3) rabbits; it can be classified as non irritating.