3,7-Diazabicyclo[3.3.1]nonane-1,5-dicarboxylic acid, 3-methyl-9-oxo-2,4-di-2-pyridinyl-7-(2-pyridinylmethyl)-, 1,5-dimethyl ester, (1R,2S,4R,5S)-rel-

Eye irritation

Administrative data

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

From 8 January 2004 to 29 March 2004

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2004

Report date:

2004

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Version / remarks:

Adopted the : 24th April 2002

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

GLP compliance:

yes

Test material

Test material information

Constituent 1

Reference substance name:

618-281-9

EC Number:

618-281-9

Cas Number:

895520-71-5

Molecular formula:

C28 H29 N5 O5

IUPAC Name:

618-281-9

Test material form:

solid: particulate/powder

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: E-699-2003
- Expiration date of the lot/batch: 1 July 2004
- Purity test date: 27 and 28 October 2003

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: white to light beige powder stored at approximately 20°C in a fume cupboard

Test animals / tissue source

Species:

rabbit

Strain:

New Zealand White

Remarks:

Crl: KBL (NZW) BR (N.Zeal.White)

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Source: Charles River, Deutschland
- Age at study initiation:Not specified
- Weight at study initiation: 3.2 to 3.8 kg
- Housing: In fully air conditioned romms in separate cages arranged in a battery
- Diet (e.g. ad libitum):ssniff K-H (V2333), ad libitum and hay (15g daily)
- Water (e.g. ad libitum): tap water in plastic bottles ad libitum
- Acclimation period: 1 week under study conditions

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20±3 deg celsius
- Humidity (%): 50±20%
- Air changes (per hr): not specified
- Photoperiod (hrs dark / hrs light): 12h/12h

IN-LIFE DATES: From: 13 January 2004 To: 23 January 2004

Test system

Vehicle:

unchanged (no vehicle)

Controls:

yes, concurrent no treatment

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 100 mg of DP-2

VEHICLE
No vehicle was used

Duration of treatment / exposure:

24 hours

Observation period (in vivo):

1 hours, 24 hours, 48 hours and 72 hours

Number of animals or in vitro replicates:

3 animals were used

Details on study design:

REMOVAL OF TEST SUBSTANCE
- Washing (if done): isotonic saline solution at 37 degree celsius
- Time after start of exposure: 24 hours

SCORING SYSTEM:
Cornea

Opacity: degree of density (readings should be taken from most dense area)*No ulceration or opacity.... 0
Scattered or diffuse areas of opacity (other than slight dulling of normal lustre); details of iris clearly visible ................ 1
Easily discernible translucent area; details of iris slightly obscured ............................................................ 2
Nacrous area; no details of iris visible; size of pupil barely discernible ...................................................... 3
Opaque cornea; iris not discernible through the opacity .............................................................................. 4

* The area of corneal opacity should be noted
Iris

Normal .......................................................................................................................................................... 0
Markedly deepened rugae, congestion, swelling, moderate circumcorneal hyperaemia;
or injection; iris reactive to light (a sluggish reaction is considered to be an effect .................................. 1
Hemorrhage, gross destruction, or no reaction to light ................................................................................. 2

Conjunctivae

Redness (refers to palpebral and bulbar conjunctivae; excluding cornea and iris)
Normal .......................................................................................................................................................... 0
Some blood vessels hyperaemic (injected) ................................................................................................... 1
Diffuse, crimson colour; individual vessels not easily discernible ............................................................... 2
Diffuse beefy red........................................................................................................................................... 3

Chemosis

Swelling (refers to lids and/or nictating membranes)
Normal .......................................................................................................................................................... 0
Some swelling above norma ......................................................................................................................... 1
Obvious swelling, with partial eversion of lids............................................................................................. 2
Swelling, with lids about half closed ............................................................................................................ 3
Swelling, with lids more than half closed ..................................................................................................... 4

TOOL USED TO ASSESS SCORE: fluorescein

Results and discussion

In vivo

Results

Results 1

Irritation parameter:

conjunctivae score

Basis:

mean

Time point:

24/48/72 h

Score:

0.44

Max. score:

1

Reversibility:

fully reversible

Remarks on result:

no indication of irritation

Results 2

Irritation parameter:

chemosis score

Basis:

mean

Time point:

24/48/72 h

Score:

0.11

Max. score:

0.33

Reversibility:

fully reversible

Remarks on result:

no indication of irritation

Results 3

Irritation parameter:

cornea opacity score

Basis:

mean

Time point:

24/48/72 h

Score:

0

Max. score:

0

Reversibility:

fully reversible

Remarks on result:

no indication of irritation

Results 4

Irritation parameter:

iris score

Basis:

mean

Time point:

24/48/72 h

Score:

0

Max. score:

0

Reversibility:

fully reversible

Remarks on result:

no indication of irritation

Irritant / corrosive response data:

From one hour up to 48 hours after administration the conjunctivae of the animals showed sporadically definitely hyperaemic blood vessels up to diffuse crimson color. Furthermore, the animals showed slight swellings one hour up to 24 hours after administration in two animals. 72 hours after administration all signs of irritation had disappeared.

Any other information on results incl. tables

Opacity of cornea	All animals	0.0	Iris	All animals	0.0
	animal 1	0.0		animal 1	0.0
	animal 2	0.0		animal 2	0.0
	animal 3	0.0		animal 3	0.0
Redness of conjunctiva	All animals	0.44	Chemosis of conjunctiva	All animals	0.11
	animal 1	1.00		animal 1	0.33
	animal 2	0.33		animal 2	0.0
	animal 3	0.0		animal 3	0.0