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

Acute toxicity: via oral route

Link to relevant study records

Reference

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

20 July to 5 November 1999

Reliability:

1 (reliable without restriction)

Qualifier:

according to guideline

Guideline:

OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method)

Deviations:

not specified

Qualifier:

according to guideline

Guideline:

EU Method B.1 tris (Acute Oral Toxicity - Acute Toxic Class Method)

Deviations:

not specified

GLP compliance:

yes

Test type:

acute toxic class method

Limit test:

yes

Species:

rat

Strain:

other: Wistar Crl:(WI) BR

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River Deutschland, Sulzfeld, Germany
- Age at study initiation: Approximately 7 weeks old
- Weight at study initiation: Within 20% of sex mean
- Fasting period before study: Overnight prior to dosing until approximately 3-4 hours after administration of the test substance.
- Housing: Group housing of 3 animals per sex per cage in polycarbonate cages containing purified sawdust as bedding material.
- Diet (e.g. ad libitum): Free access to standard pelleted laboratory animal diet (Carfil Quality BVBA, OUd-Turnhour, Belgium).
- 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 Air changes per hour
- Photoperiod (hrs dark / hrs light): 12 hours of artificial fluorescent light/12 hours dark.

Route of administration:

oral: gavage

Vehicle:

polyethylene glycol

Details on oral exposure:

VEHICLE
- Amount of vehicle (if gavage): 10mL/kg
- Justification for choice of vehicle: The vehicle was selected based on a pretest performed at NOTOX.

DOSAGE PREPARATION (if unusual): Formulations (w/w) were prepared within 4 hours prior to dosing. Homogeneity was accomplished to a visually acceptable level.

CLASS METHOD (if applicable)
- Rationale for the selection of the starting dose: Toxicity of the test substance was assessed by stepwise treatment of groups of 3 animals.

Doses:

200 mg/kg
2000 mg/kg

No. of animals per sex per dose:

3 Animals

Control animals:

not specified

Details on study design:

- Duration of observation period following administration: 15 days
- Frequency of observations and weighing: Twice daily
- Necropsy of survivors performed: Yes, by asphyxiation using oxygen/carbon dioxide procedure.
- Other examinations performed: clinical signs, body weight, organ weights, histopathology, other: Twice daily viability/mortality; body weights Days 1 (pre-admin), 8 and 15 and at death; clinical signs were taken on day 1 and once daily thereafter, until day 15.

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 200 - < 2 000 mg/kg bw

Based on:

test mat.

Mortality:

The incidence of mortality is reported in Table No. 1 below.

Two females and two males were found dead on days 2 and 4 and on days 4 and 5, respectively. The two other animals (one female and one male) were sacrificed moribund on day 7 and day 5, respectively, as indicated by the combination of clinical signs and considerable body weight loss observed in these animals.

Clinical signs:

other: Within the 200 mg/kg dose group, signs of alopecia and/or scabs (in the neck) became apparent in two females during week 2. Based on the time of occurrence, these findings were considered not related to treatment and of no toxicological significance. Wi

Gross pathology:

Reduced spleen and thymus size was found among the 2000 mg/kg treated animals, at macroscopic post mortem examination.

Table 1:

Dose Level (mg/kg)	Mortality	Sex	Date of Treatment
200	0/3	females	20-Jul-99
200	0/3	males	22-Jul-99
2000	3/3	females	27-Jul-99
2000	3/3	males	29-Jul-99

Interpretation of results:

harmful

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

Under the current test conditions the oral LD50 value for the test substance is within the following range 200-2000 mg/kg body weight.

Endpoint conclusion

Endpoint conclusion:

adverse effect observed

Dose descriptor:

LD50

Value:

200 mg/kg bw

Acute toxicity: via inhalation route

Endpoint conclusion

Endpoint conclusion:

no study available

Acute toxicity: via dermal route

Link to relevant study records

Reference

Endpoint:

acute toxicity: dermal

Type of information:

experimental study

Adequacy of study:

key study

Study period:

12 August to 3 November 1999

Reliability:

1 (reliable without restriction)

Qualifier:

according to guideline

Guideline:

OECD Guideline 402 (Acute Dermal Toxicity)

Deviations:

not specified

Qualifier:

according to guideline

Guideline:

EU Method B.3 (Acute Toxicity (Dermal))

Deviations:

not specified

GLP compliance:

yes

Test type:

standard acute method

Limit test:

no

Species:

rat

Strain:

other: Wistar Crl:(WI) BR

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River Deutschland, Sulzfeld, Germany
- Age at study initiation: Approx. 9 weeks old
- Weight at study initiation: Did not exceed +/- 20% of sex mean.
- Housing: Individually housed in polycarbonated cages containing purified saw dust as bedding material.
- Diet (e.g. ad libitum): Free access to standard pelleted laboratory animal diet (Carfil Quality BVBA, Oud-Turnout, Belgium).
- 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): Approximately 15 air changes per hour.
- Photoperiod (hrs dark / hrs light): 12 hours of artificial light/12 hours dark

IN-LIFE DATES: From: 12 August 1999 To: 26 August 1999

Type of coverage:

semiocclusive

Vehicle:

polyethylene glycol

Details on dermal exposure:

TEST SITE
- Area of exposure: Back of animals
- % coverage: 10%
- Type of wrap if used: Surgical gauze patch, successively covered with aluminium foil and Coban flexible bandage. A piece of Micropore tape was additionally used for fixation of the bandages to females only.

REMOVAL OF TEST SUBSTANCE
- Washing (if done): Water
- Time after start of exposure: 24 hours

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 10 mL/kg body weight

Duration of exposure:

24 hours

Doses:

2000 mg/kg

No. of animals per sex per dose:

5 Males and 5 Females

Control animals:

not required

Details on study design:

- Duration of observation period following administration: 15 days
- Frequency of observations and weighing: Observations - twice daily, body weights - days 1 (pre-administration), 8 and 15.
- Necropsy of survivors performed: Yes, Sacrificed by asphyxiation using an oxygen/carbon dioxide procedure.
- Other examinations performed: clinical signs, body weight, organ weights, histopathology, other: Clinical signs - On day 1 of dosing and once daily thereafter, until day 15.

Statistics:

Not performed

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Mortality:

No mortality occurred

Clinical signs:

other: Scabs and scales were seen in the treated skin-area of one female between days 2 and 8. Red staining of the fur in the neck appeared in another female on day 9 and persisted until termination. Based on the incidence and time of occurrence, this finding

Gross pathology:

No abnormalities were found at macroscopic post mortem examination of the animals.

Interpretation of results:

practically nontoxic

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

Under current test conditions the LD50 value for test substance exceeds 2000 mg/kg body weight.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

LD50

Value:

2 000 mg/kg bw

Additional information

Justification for selection of acute toxicity – oral endpoint
GLP study conducted according to OECD Guideline 423 and EU Method B.1. Under the current test conditions the oral LD50 value for Pymordiol is within the following range 200-2000 mg/kg body weight.

Justification for selection of acute toxicity – dermal endpoint
CLP study conducted according to OECD Guideline 402 and EU Method B.3. Under current test conditions the LD50 value for test substance exceeds 2000 mg/kg body weight.

Justification for classification or non-classification

Based on acute oral LD50 greater than 200 but less than 2000 mg/kg bw, this substance might either be classified as EU CLP Acute Oral Tox Cat 3 (if the LD50 is less than 300 mg/kg bw) or Acute Oral Tox Cat 4 (if LD50 is greater than 300 mg/kg bw).

Precautionarily, this substance is therefore classified as Acute Oral Tox Cat 3, H301.