.beta.-D-Galactopyranoside, ethyl 3-O-[(1S)-1-(cyclohexylmethyl)-2-oxo-2-(phenylmethoxy)ethyl]-1-thio-, 2,4,6-tribenzoate

Eye irritation

Administrative data

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

28 September - 12 November 2015

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2015

Report date:

2016

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Test guideline 2

Qualifier:

according to guideline

Guideline:

other: EC 440/2008, B5: "Acute Toxicity: Eye Irritation/Corrosion"

Test guideline 3

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.2400 (Acute Eye Irritation)

Test guideline 4

Qualifier:

according to guideline

Guideline:

other: JMAFF Guidelines (2000), including the most recent revisions

GLP compliance:

yes

Test material

Test material information

Constituent 1

Reference substance name:

(2S,3R,4S,5S,6R)-5-(benzoyloxy)-6-[(benzoyloxy)methyl]-4-{[(2S)-1-(benzyloxy)-3-cyclohexyl-1-oxopropan-2-yl]oxy}-2-(ethylsulfanyl)oxan-3-yl benzoate

EC Number:

805-702-8

Cas Number:

322761-81-9

Molecular formula:

C45H48O10S

IUPAC Name:

(2S,3R,4S,5S,6R)-5-(benzoyloxy)-6-[(benzoyloxy)methyl]-4-{[(2S)-1-(benzyloxy)-3-cyclohexyl-1-oxopropan-2-yl]oxy}-2-(ethylsulfanyl)oxan-3-yl benzoate

Test material form:

solid: particulate/powder

Test animals / tissue source

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Source: Charles River France, L’Arbresle Cedex, France.
- Age at study initiation: 3 males, at start of dosing, the animals were between 12 and 24 weeks old.
- Weight at study initiation: Body weights were at leat 1.5 kg.
- Housing: Animals were individually housed in labeled cages with perforated floors (Ebeco, Germany,
dimensions 67 x 62 x 55 cm) and shelters (Ebeco, Germany, dimensions 40 x 32 x 23 cm).
- Diet: Pelleted diet for rabbits (Global Diet 2030 from Harlan Teklad®, Mucedola, Milanese, Italy)
approximately 100 grams per day. Hay (TecniLab-BMI BV, Someren, The Netherlands) and wooden
sticks (Swedish aspen wood, Bioservices, Uden, The Netherlands) were available during the study
period.
- Water: Free access to tap water.
- Acclimation period: Acclimatization period was at least 5 days before start of treatment under laboratory conditions.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 18 to 24°C
- Humidity (%): A relative humidity of 40
to 70%
- Air changes (per hr): At least 10 air changes/hour
- Photoperiod (hrs dark / hrs light): A 12-hour light/12-hour dark cycle

IN-LIFE DATES: From: 28 September 2015 To: 12 November 2015

Test system

Vehicle:

unchanged (no vehicle)

Controls:

yes, concurrent no treatment

Amount / concentration applied:

Each animal will be treated by instillation of a volume of 0.1 mL of the test item* in the conjunctival sac
of one of the eyes after gently pulling the lower lid away from the eyeball. The lids will then be gently
held together for approximately one second to prevent loss of test item. The non-treated eye will serve
as the control.
If a solid, the weight will be recorded. A maximum of approximately 100 mg will be instilled.

Duration of treatment / exposure:

Immediately after completion of the 1 hour examination the test eye of each animal may be rinsed with tap water to remove residual test item visually present. For reference control, the other eye will also be rinsed.

Observation period (in vivo):

Immediately after the 24 hour examination, both eyes of each animal will be further examined with the aid of fluorescein stain. Any bright green stained area, indicating epithelial damage, will be estimated as a percentage of the total corneal area. If epithelial damage is observed this procedure will be repeated at later observation times, to assess recovery.
Following the completion of the 24 hour examination the test eye of each animal may be rinsed with tap water to remove residual test item visually present. For reference control, the other eye will also be rinsed.
The eyes of each animal will be examined approximately 1, 24, 48 and 72
hours after administration of the test item. At least for the duration of the
irritation (including local effects and staining of the eye; criteria are
described in WIL Research Europe Standard Operating Procedure DIE
J/125)), further observations will be made 7, 14 and 21 (maximum) days
after administration. The irritation scores and a description of all other
(local) effects will be recorded.

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 six weeks later, after considering
the degree of eye irritation observed in the first animal.
Observations:
Mortality/Viability: At least twice daily. Special attention will be paid for signs of pain, distress
or discomfort in order to decide in a timely manner on (i) the application of
systemic analgesics and (ii) the sacrifice of the animals for humane
reasons if these signs are not transient in nature or are likely to become
more severe (based on OECD guidance document on humane endpoints
(ENV/JM/MONO/ 2000/7)). Decisions will be documented in the study
files.
Toxicity At least once daily. If toxicity is observed, all clinical signs will be recorded
until they have disappeared.
Body Weight Measured on Day 1 (prior to treatment) and after the final observation or
at death (if found dead after Day 1).

Results and discussion

In vivo

Results

Results 1

Irritation parameter:

cornea opacity score

Basis:

mean

Time point:

24/48/72 h

Score:

0

Max. score:

0

Reversibility:

other: No irritation noted

Results 2

Irritation parameter:

iris score

Basis:

mean

Time point:

24/48/72 h

Score:

0

Max. score:

0

Reversibility:

other: No irritation noted

Results 3

Irritation parameter:

conjunctivae score

Basis:

mean

Time point:

other: 1 hr

Score:

4.7

Max. score:

6

Reversibility:

fully reversible within: 48 hrs

Remarks on result:

probability of mild irritation

Results 4

Irritation parameter:

conjunctivae score

Basis:

mean

Time point:

24 h

Score:

0.7

Max. score:

2

Reversibility:

fully reversible within: 48 hrs

Remarks on result:

probability of mild irritation

Results 5

Irritation parameter:

chemosis score

Basis:

mean

Time point:

24/48/72 h

Score:

0

Max. score:

0

Reversibility:

other: No irritation noted

Results 6

Irritation parameter:

overall irritation score

Basis:

mean

Time point:

24/48/72 h

Score:

0

Max. score:

0

Reversibility:

other: No irritation noted

Results 7

Irritation parameter:

maximum mean total score (MMTS)

Basis:

mean

Time point:

other: 1 hr

Score:

4.7

Max. score:

6

Reversibility:

fully reversible within: 48 hrs.

Remarks on result:

probability of mild irritation

Results 8

Irritation parameter:

maximum mean total score (MMTS)

Basis:

mean

Time point:

24 h

Score:

0.7

Max. score:

2

Reversibility:

fully reversible within: 48 hrs

Remarks on result:

probability of mild irritation

Irritant / corrosive response data:

Instillation of approximately 38 mg of PF-06460246 (a volume of approximately 0.1 mL) into one eye of each of three rabbits resulted in irritation of the conjunctivae, which consisted of redness and discharge. The irritation had completely resolved within 24 hours in two animals and within 48 hours in the other animal.
No iridial irritation or corneal opacity was observed, and treatment of the eyes with 2% fluorescein 24 hours after test item instillation revealed no corneal epithelial damage.
There was no evidence of ocular corrosion.
Remnants of the test item were present in the eye of all animals on Day 1.
No signs of systemic toxicity were observed in the animals during the test period and no mortality occurred.

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

Based on these results, PF-06460246 does not have to be classified and has no obligatory labelling
requirement for eye irritation according to the:
- Globally Harmonized System of Classification and Labelling of Chemicals (GHS) of the United
Nations (2011) (including all amendments)

Executive summary:

Based on the maximum group mean of 4.7, PF-06460246 is classified as minimal irritant to the rabbit eye according to the Kay and Calandra classification system.