2,2,4,4-tetramethyl-7-oxa-3,20-diazadispiro[5.1.11.2]henicosan-21-one hydrochloride

Eye irritation

Administrative data

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2008-06-13 till 2008-07-17

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: guideline-conform study under GLP without deviations

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

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

2,2,4,4-tetramethyl-7-oxa-3,20-diazadispiro[5.1.11.2]henicosan-21-one hydrochloride

EC Number:

277-620-5

EC Name:

2,2,4,4-tetramethyl-7-oxa-3,20-diazadispiro[5.1.11.2]henicosan-21-one hydrochloride

Cas Number:

73833-37-1

Molecular formula:

C22H40N2O2.xClH

IUPAC Name:

2,2,4,4-tetramethyl-7-oxa-3,20-diazadispiro[5.1.11.2]henicosan-21-one hydrochloride

Details on test material:

- Name of test material (as cited in study report): 2,2,4,4-tetramethyl-7-oxa-3,20-diazadispiro[5.1.11.2]henicosan-21-one hydrochloride
- Physical state: solid, white
- Analytical purity: => 99%
- Lot/batch No.: DEF2036584
- Expiration date of the lot/batch: 2018-05-08
- Stability under test conditions: stable
- Storage condition of test material: At room temperature (range of 20 ± 5 °C), light protected.

Test animals / tissue source

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Source: Harlan Netherlands BV
- Age at study initiation: 15 weeks (male); 16-17 weeks (females)
- Weight at study initiation: 2767 g (male); 2677 g and 3033 g (females)
- Housing: Individually in stainless steel cages equipped with feed hoppers and drinking water bowls.
- Diet (e.g. ad libitum): Pelleted standard Provimi Kliba 3418 rabbit maintenance diet ad libitum
- Water (e.g. ad libitum): Community tap water from Füllinsdorf ad libitum
- Acclimation period: 2008-06-13 to 2008-06-17


ENVIRONMENTAL CONDITIONS
- Temperature (°C): 17 - 23°C
- Humidity (%): 30 - 70%
- Air changes (per hr): 10 - 15
- Photoperiod (hrs dark / hrs light):12 / 12

Test system

Vehicle:

unchanged (no vehicle)

Controls:

other: control area on same animal (right eye)

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.1 g per animal

Duration of treatment / exposure:

72 hrs

Observation period (in vivo):

1, 24, 48, 72 hours as well as 7, 10, 14, 17, 21 and 28 days after administration.

Number of animals or in vitro replicates:

3 (one male, two females)

Details on study design:

REMOVAL OF TEST SUBSTANCE
- Washing (if done): The treated eye of each animal was rinsed with NaCl 0.9% after each assessment from test days 2 to 18 due to sticky test item remnants in the eye.


SCORING SYSTEM: numerical scoring system listed in the COUNCIL REGULATION (EC) No 440/2008 of 30 May 2008

Results and discussion

In vivo

Results

Results 1

Irritation parameter:

cornea opacity score

Basis:

mean

Time point:

other: 24 h, 48 h, 72 h

Score:

>= 1 - <= 1.67

Max. score:

1.67

Reversibility:

not fully reversible within: 28 days

Results 2

Irritation parameter:

cornea opacity score

Basis:

mean

Time point:

other: 7, 10, 14, 17, 21 and 28 days

Score:

>= 0.33 - <= 2.17

Max. score:

2.17

Reversibility:

not fully reversible within: 28 days

Results 3

Irritation parameter:

iris score

Basis:

mean

Time point:

other: 24 h, 48 h, 72 h

Score:

1

Max. score:

1

Reversibility:

fully reversible within: 17 days

Results 4

Irritation parameter:

iris score

Basis:

mean

Time point:

other: 7, 10, 14, 17, 21 and 28 days

Score:

>= 0 - <= 0.5

Max. score:

0.5

Reversibility:

fully reversible within: 17 days

Results 5

Irritation parameter:

conjunctivae score

Basis:

mean

Time point:

other: 24 h, 48 h, 72 h

Score:

3

Max. score:

3

Reversibility:

not fully reversible within: 28 days

Results 6

Irritation parameter:

conjunctivae score

Basis:

mean

Time point:

other: 7, 10, 14, 17, 21 and 28 days

Score:

>= 0.5 - <= 1

Max. score:

1

Reversibility:

not fully reversible within: 28 days

Results 7

Irritation parameter:

chemosis score

Basis:

mean

Time point:

other: 24 h, 48 h, 72 h

Score:

>= 2.67 - <= 4

Max. score:

4

Reversibility:

not fully reversible within: 28 days

Results 8

Irritation parameter:

chemosis score

Basis:

mean

Time point:

other: 7, 10, 14, 17, 21 and 28 days

Score:

>= 0.33 - <= 1.67

Max. score:

1.67

Reversibility:

not fully reversible within: 28 days

Results 9

Irritation parameter:

other: sclera

Basis:

mean

Time point:

other: 24 h, 48 h, 72 h, 7 d, 10 d, 14 d, 17 d, 21 d and 28 d

Score:

3

Max. score:

3

Reversibility:

fully reversible within: 28 days

Any other information on results incl. tables

Table 1: Irritant/corrosive response data for each animal at each observation time up to removal of each animal from the test

Score at time point / Reversibility	Cornea	Iris	Conjunctivae	Chemosis	Sclera
	Max. score: 3	Max. score: 1	Max. score: 3	Max. score: 4	Max. score: 3
60 min	0/0/0	0/0/0	1/1/1	1/1/1	1/1/2
24 h	1/1/1	1/1/1	3/3/3	4/4/4	n.a.
48 h	2/1/1	1/1/1	3/3/3	4/2/2	n.a./3/3
72 h	2/1/1	1/1/1	3/3/3	4/2/2	n.a./3/3
Average 24h, 48h, 72h	1.67/1/1	1/1/1	3/3/3	4/2.67/2.67
Maximum average score (including area affected, max 110)	1.67	1	3	4
7 days	3/2/1	1/0/0	3/2/2	2/1/1	3/2/1
10 days	3/2/1	1/0/0	3/2/2	2/1/1	3/2/1
14 days	3/1/0	1/1/0	2/1/0	2/1/0	3/1/0
17 days	2/1/0	0/0/0	2/1/0	2/1/0	2/1/0
21 days	1/1/0	0/0/0	1/0/0	1/0/0	1/0/0
28 days	1/0/0	0/0/0	1/0/0	1/0/0	0/0/0
Area effected	4	-	-	-	-
Reversibility*)	n.c.	c	n.c.	n.c.	c
Average time (unit) for reversion	-	17 days	-	-	28 days

*) Reversibility: c. = completely reversible; = not completely reversible; n. = not reversible

Applicant's summary and conclusion

Interpretation of results:

highly irritating

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

67/548/EEC: R41
1272/2008/EC:Irreversible effects on the eye ( Category 1) / H318

Executive summary:

The primary irritation potential of Hostavin N 20 - Hydrochlorid was investigated according to OECD test guideline 405.

A delayed/reduced light reflex of the iris was noted in all animals from the 24 hour observation until the 72-hour reading and in the male on days 7, 10 and 14 after treatment and in one female again on day 14 after tretament.

A slight reddening of the conjunctivae was noted 1 hour after treatment in all animals and increased to marked 24 hours post treatment with bleeding at the conjunctivae on test day 2 in the male. A marked redness persisted up to the 72-hour observation and, thereafter decreased to

moderate and slight up to 28 days post treatment in the male. Slight swelling (chemosis) of the conjunctivae was present at the 1-hour reading in all animals. A marked swelling (with half-closed to closed lids) was noted at the 24-hour reading in all animals. The swelling decreased to an obvious swelling with partial eversion of lids and slight swelling 48 hours after treatment or on day 7 post treatment and persisted up to days 10, 21 or 28 post treatment.

A slight to moderate reddening of the sclerae was present in all animals at the 1-hour reading. At 24 hours assessment of the sclera was not assessable in all animals due to the marked swelling of the conjunctivae. A marked reddening was noted at the 48- and 72-hour reading in the two females. In the male the sclerae was still not assessable due to the swelling of conjunctivae. On days 7 and 10 post treatment a slight to marked reddening was recorded that persisted in two animals up to day 17 and in the male to test day 21 after treatment.

A very slight opacity affecting the whole area of the cornea was observed at the 24-hour reading in all animals and persisted as very slight to moderate (affecting the whole area) in all animals until day 10 post treatment. This persisted decreasing to very slight in two animals up to day 21 and in the male (greater than one quarter but less than the half corneal area) up to day 28 post treatment.

Neovascularisation as well as Plaques (size: a few millimiters) was noted in the cornea in all animals on days 22 (additional observation) and 28 post treatment.

Slight ocular discharge was visible 1 hour after instillation in all animals, increased to marked at 24 hours and persisted as such up to the 72-hour reading. Afterwards, slight to moderate discharge was noted in all animals up to days 10 and, further on up to days 14 and/or 17 after treatment. Mucus was present in all animals 24 to 72 hours post treatment and in one animal again at the

10-day observation.

White test item remnants were evident in the treated eyes of all three animals at the 1-hour reading and again from the 48-hour reading up to day 14 in all animals and up to day 17 post tretament in one animal. Due to mucus / discharge test item remnants were not assessable on the 24 -hour reading.

No staining produced by the test item of the treated eye was observed. No corrosion of the cornea was observed at any of the reading times. One female did not gain weight between the first day of acclimatization and the day of treatment, but recovered until the end of the study. Otherwise, the body weights of all rabbits were considered to be within the normal range of variability.