Androstane-3,17-diol, 2-(4-morpholinyl)-16-(1-pyrrolidinyl)-, (2.beta.,3.alpha.,5.alpha.,16.beta.,17.beta.)-

Endpoint summary

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:

31 August to 5 November 1999

Reliability:

1 (reliable without restriction)

Qualifier:

according to guideline

Guideline:

OECD Guideline 404 (Acute Dermal Irritation / Corrosion)

Deviations:

not specified

Qualifier:

according to guideline

Guideline:

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

Deviations:

not specified

GLP compliance:

yes

Species:

rabbit

Strain:

New Zealand White

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River Nederland, Someren, the Netherlands
- Age at study initiation: 6 weeks
- Weight at study initiation: Less than 3.5 kg
- Housing: Individually in cages with perforated floors.
- Diet: Approximately 100 g per day of standard laboratory rabbit diet (Charles River Breeding and Maintenance Diet for Rabbits, Altromin, Germany). Hay (BMI, Helmond, the Netherlands) was provided once a week.
- Water (e.g. ad libitum): Free-access to tap water.
- Acclimation period: 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21˚C
- Humidity (%): 50%
- Air changes (per hr): 15 changes per hour
- Photoperiod (hrs dark / hrs light): 12 hours artificial fluorescent light/ 12 hours dark

IN-LIFE DATES: From: To:

Type of coverage:

semiocclusive

Preparation of test site:

shaved

Vehicle:

water

Controls:

not required

Amount / concentration applied:

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

Duration of treatment / exposure:

4 hours

Observation period:

72 hours

Number of animals:

3 of single sex

Details on study design:

TEST SITE
- Area of exposure: Dorsal flank
- % coverage: 150 square centimetres
- Type of wrap if used: Metalline patch of 2x3 cm. Patch was mounted on Micropore tape, which was wrapped around the abdomen and secured with Coban elastic bandage.

REMOVAL OF TEST SUBSTANCE
- Washing (if done): Water
- Time after start of exposure: 4 hours

SCORING SYSTEM: See table below

Irritation parameter:

overall irritation score

Basis:

mean

Time point:

other: 72 hours

Score:

ca. 0

Max. score:

4

Irritant / corrosive response data:

No evidence of irritation after 4 hours of exposure to test substance was reported.

See tables No. 1 & 2 below for details on individual animals and animal means.

Other effects:

No staining of the treated skin by the test substance was observed. 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 No.	494		496		518
Time after exposure (hours)	Erythema	Oedema	Erythema	Oedema	Erythema	Oedema
1	0	0	0	0	0	0
24	0	0	0	0	0	0
48	0	0	0	0	0	0
72	0	0	0	0	0	0

Table 2: Mean Values of Skin Irritation Scores (24, 48 and 72 h after exposure):

Animal No.	Mean 24-72 hours
	Erythema	Oedema
494	0	0
496	0	0
518	0	0

Interpretation of results:

not irritating

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

Under the current test conditions test substance is not considered to be irritating to the skin.

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:

6 September to 5 November 1999

Reliability:

1 (reliable without restriction)

Qualifier:

according to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Deviations:

not specified

Qualifier:

according to guideline

Guideline:

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

Deviations:

not specified

GLP compliance:

yes

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Source: Charles River Deutschland, Kisslegg, Germany
- Age at study initiation: At least 6 weeks of age
- Weight at study initiation: 3.5 kg
- Housing: Individually cages with perforated floors
- Diet: 100 g per day of Standard laboratory rabbit diet (Charles River Breeding and Maintenance Diet for Rabbits, Altromin, Germany). Hay was provided once per week (BMI, Helmond, the Netherlands).
- Water (e.g. ad libitum): Free access to tap water
- Acclimation period: 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21˚C
- Humidity (%): 50%
- Air changes (per hr): 15 changes per hour
- Photoperiod (hrs dark / hrs light): 12 hours artificial fluorescent light/ 12 hours dark

Vehicle:

unchanged (no vehicle)

Controls:

not required

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 54.6 mg +/- 0.4 mg (volume of approx. 0.1 mL)

Duration of treatment / exposure:

24 hours

Observation period (in vivo):

72 hours

Number of animals or in vitro replicates:

3 Animals of one sex

Details on study design:

SCORING SYSTEM: See Tables No. 1 - 5 below

TOOL USED TO ASSESS SCORE: Fluorescein, and the use of ophthalmic examination lamp

Irritation parameter:

cornea opacity score

Basis:

mean

Time point:

other: 24-72 hours

Score:

ca. 0

Max. score:

4

Irritation parameter:

iris score

Basis:

mean

Time point:

other: 24-72 hours

Score:

ca. 0

Max. score:

2

Irritation parameter:

conjunctivae score

Remarks:

Redness

Basis:

mean

Time point:

other: 24-72 hours

Score:

ca. 0.67

Max. score:

3

Reversibility:

fully reversible within: 72 hours

Irritation parameter:

chemosis score

Basis:

mean

Time point:

other: 24-72 hours

Score:

ca. 0

Max. score:

4

Irritant / corrosive response data:

Iridial irritation was reported in all animals on day 1 and was resolved within 24 hours.

Irritation of the conjunctivae was seen as redness, chemosis and discharge, which was completely resolved within 48 hours in one animal and 72 hours in the other test animals.

No corneal opacity was reported.

See tables No. 6 - 7 below for details on individual test animals and mean scores.

Other effects:

No symptoms of systemic toxicity was observed in the animals during the test period and no mortality was observed.

Table 6: Individual Eye Irritation Scores

Animal No.			469			470			471
Time After Dosing	Tissue/Finding		Score	Fluor. Area (%)	Comments	Score	Fluor. Area (%)	Comments	Score	Fluor. Area (%)	Comments
1 hour	Corneal	Opacity	0		Remnants of test substance in the eye	0		Remnants of test substance in the eye	0		Remnants of test substance in the eye
		Area	0			0			0
	Iris		1			1			1
	Conj	Redness	1 (ABC)			2 (C)			2(C)
		Chemosis	1 (B)			1 (ABC)			2(C)
	Discharge		1			1			1
24 hours	Corneal	Opacity	0	0		0			0
		Area	0			0			0
	Iris		0			0			0
	Conj	Redness	1 (ABC)			2 (C)			1 (ABC)
		Chemosis	0			0			0
	Discharge		0			1			0
48 hours	Corneal	Opacity	0			0			0
		Area	0			0			0
	Iris		0			0			0
	Conj	Redness	0			1(C)			1(C)
		Chemosis	0			0			0
	Discharge		0			0			0
72 hours	Corneal	Opacity	0			0			0
		Area	0			0			0
	Iris		0			0			0
	Conj	Redness	0			0			0
		Chemosis	0			0			0
	Discharge		0			0			0

Score given for:

A = Eyelids,

B = Nictitating membrane, and

C = Sclera

Table 7: Mean Values of Eye Irritation Scores (24 -72 h after instillation)

Animal No.	Mean 24 - 72 hours
	Corneal Opacity	Iris	Conjunctivae
			Redness	Chemosis
469	0	0	0.3	0
470	0	0	1	0
471	0	0	0.7	0

Interpretation of results:

not irritating

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

Under current test conditions test substance is not considered irritating to the eye.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Respiratory irritation

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

Justification for selection of skin irritation / corrosion endpoint:
GLP study conducted according to OECD Guideline 404 and EU Method B.4. Under the current test conditions test substance is not considered to be irritating to the skin.

Justification for selection of eye irritation endpoint:
GLP study conducted according to OECD Guideline 405 and EU Method B.5. Under current test conditions test substance is not considered irritating to the eye.

Justification for classification or non-classification

Pymordiol does not meet the classification criteria for irritation and is therefore not classified as a skin or eye irritant according to EU CLP and EU DSD.