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

Endpoint summary

Description of key information

Skin irritation:

The test substance was applied to the skin of three rabbits (OECD 404 & EC B4) for four hours then washed off. No skin irritation was caused by 4 hours exposure to the test substance in three (3/3) rabbits. The substance can be considered non irritant to the skin.  

Eye irritation:

The test substance was also applied to the conjunctival sac of one of the eyes of each of three male rabbits (OECD 405 & EC B5). Treatment with the test substance caused reversible effects on the cornea, iris and conjunctivea in the eye of three (3/3) rabbits; it can be classified as non irritating.

Key value for chemical safety assessment

Skin irritation / corrosion

Link to relevant study records

Reference

Endpoint:

skin irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

29 January 2008 - 01 February 2008

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Study performed to OECD, European and other guidelines and statement of GLP compliance is included in report.

Qualifier:

according to guideline

Guideline:

OECD Guideline 404 (Acute Dermal Irritation / Corrosion)

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method B.4 (Acute Toxicity: Dermal Irritation / Corrosion)

Deviations:

no

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.2500 (Acute Dermal Irritation)

Deviations:

no

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

Species:

rabbit

Strain:

New Zealand White

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Harlan, Horst, The Netherlands
- Age at study initiation: One animal used within the study was 9 to 11 weeks old and the two other animals were at least 11 weeks old.
- Weight at study initiation: At least 2.0 kg.
- Housing: Individually in labelled cages with perforated floors (Scanbur, Denmark, dimensions 56x44x37.5 cm).
- Diet (e.g. ad libitum): Pelleted diet for rabbits (K-H from SSNIFF® Spezialdiaten GmbH, Soest, Germany) approximately 100 grams per day. Hay
(TecniLab-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 start of treatment under laboratory conditions.


ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21.0 ± 3.0°C (actual range: 18.3 - 21.4°C)
- Humidity (%): relative humidity of 30 - 70% (actual range: 38 - 56%)
- 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.


IN-LIFE DATES: From: 29 January 2008 To: 01 February 2008

Type of coverage:

semiocclusive

Preparation of test site:

shaved

Vehicle:

other: Test substance moistened with water immediately before application.

Controls:

no

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.5 g

VEHICLE
Water (Elix, Millipore S.A.S., Molsheim, France)
- Amount(s) applied (volume or weight with unit): 0.7 mL.

Duration of treatment / exposure:

Test item was applied, them removed (washed off) after fours hours.

Observation period:

Skin reactions were assessed at approximately 1, 24, 48 and 72 hours after the removal of the dressings and test substance.

Number of animals:

3 males.

Details on study design:

TEST SITE
- Area of exposure: 2 x 3 cm (6 cm²)
- % coverage: Not specified.
- Type of wrap if used: Metalline patch (Lohmann GmbH, Neuwied, Germany) mounted on Micropore tape which was wrapped around the abdomen and secured with Coban elastic bandage (Micropore and Coban - 3M, St. Paul, Minnesota, USA).


REMOVAL OF TEST SUBSTANCE
Four hours after the application, the dressing was removed and the skin cleaned of residual test substance using tap water.

SCORING SYSTEM:

Erythema and eschar formation:
No erythema 0
Very slight erythema (barely perceptible) 1
Well-defined erythema 2
Moderate to severe erythema 3
Severe erythema (beet redness) 4
* Where signs of necrosis or corrosion (injuries in depth) prevent erythema scoring, the maximum grade for erythema (= 4) is given.

Oedema formation:
No oedema 0
Very slight oedema (barely perceptible) 1
Slight oedema (edges of area well-defined by definite raising) 2
Moderate oedema (raised approximately 1 millimeter) 3
Severe oedema (raised more than 1 millimeter and extending beyond the area of exposure) 4

Irritation parameter:

erythema score

Basis:

animal: All animals

Time point:

other: 1, 24, 48 and 72 hours

Score:

0

Max. score:

0

Irritation parameter:

other: Oedema

Basis:

animal: All animals

Time point:

other: 1, 24, 48 and 72 hours

Score:

0

Max. score:

0

Irritant / corrosive response data:

No skin irritation was caused by 4 hours exposure to LACTOL. There was no evidence of a corrosive effect on the skin.

Other effects:

Colouration / Remnants
No staining of the treated skin by the test substance was observed and no test substance remnants were seen.
Toxicity / Mortality
No symptoms of systemic toxicity were observed in the animals during the test period and no mortality occurred.

Table 1: Individual skin irritation scores

Animal	684^1			711			713
Time after exposure	Erythema	Oedema	Comments	Erythema	Oedema	Comments	Erythema	Oedema	Comments
1 hour	0	0		0	0		0	0
24 hours	0	0		0	0		0	0
48 hours	0	0		0	0		0	0
72 hours	0	0		0	0		0	0

¹Sentinel

Refer also to figure 1, attached.

Interpretation of results:

not irritating

Remarks:

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

Conclusions:

Treatment with LACTOL caused no skin irritation in three (3/3) rabbits.

Executive summary:

A Skin Irritation study was performed by NOTOX B.V. on behalf of Pfizer Ireland Pharmaceuticals, Ltd to assess the potential for skin irritation or corrosion of the test substance LACTOL. The study was performed to GLP and according to OECD guideline 404, EC guideline B4, and aditional US and Japanese guidelines.

Test substance (0.5 g) was applied to the skin of three rabbits for four hours then washed off. Skin reactions were assessed 1, 24, 48 and 72 hours after removal of the test substance.

No skin irritation was caused by 4 hours exposure to LACTOL in three (3/3) rabbits. The substance can be considered non irritant.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Eye irritation

Link to relevant study records

Reference

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.

Qualifier:

according to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Deviations:

no

Qualifier:

according to guideline

Guideline:

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

Deviations:

no

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.2400 (Acute Eye Irritation)

Deviations:

no

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

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

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.

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.

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).

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.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Additional information

Skin irritation:

The test substance was applied to the skin of three rabbits (OECD 404 & EC B4) for four hours then washed off. Skin reactions were assessed 1, 24, 48 and 72 hours after removal of the test substance. No skin irritation was caused by 4 hours exposure to the test substance in three (3/3) rabbits. The substance can be considered non irritant to the skin.

Eye irritation:

The test substance was also applied to the conjunctival sac of one of the eyes of each of three male rabbits (OECD 405 & EC B5) , and observations taken after 1, 24, 48, and 72 hours, and after 7 days. Irritation of the conjunctivae was observed, 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 the test substancecaused reversible effects on the cornea, iris and conjunctivea in the eye of three (3/3) rabbits; it can be classified as non irritating.

Justification for selection of skin irritation / corrosion endpoint:

Study performed to OECD, European and other guidelines and statement of GLP compliance is included in report.

Justification for selection of eye irritation endpoint:

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

Justification for classification or non-classification

According to the results of the tests and the Official Journal of the European Communities, Annex VI, 'General Classification and Labelling Requirements for Dangerous Substances and Preparations' the substance is not classified on the basis of its irritancy/corrosivity to the skin or to the eye.