Insulin Aspart Precursor

Acute Toxicity: dermal

Administrative data

Endpoint:

acute toxicity: dermal

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2000.03.03 - 2000.07.04.

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2000

Report date:

2010

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 402 (Acute Dermal Toxicity)

Deviations:

yes

Remarks:

Storage temperature was < -10 oC. The animals were transferred from another study and had a longer acclimatization period so they approximated to the protocol age range. The changes had no influences on the integrity or the results of the present study

Test guideline 2

Qualifier:

according to guideline

Guideline:

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

Deviations:

yes

Remarks:

Storage temperature was < -10 oC. The animals were transferred from another study and had a longer acclimatization period so they approximated to the protocol age range. The changes had no influences on the integrity or the results of the present study

Test guideline 3

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.1200 (Acute Dermal Toxicity)

Deviations:

yes

Remarks:

Storage temperature was < -10 oC. The animals were transferred from another study and had a longer acclimatization period so they approximated to the protocol age range. The changes had no influences on the integrity or the results of the present study

GLP compliance:

yes (incl. QA statement)

Test type:

fixed dose procedure

Limit test:

yes

Test material

Test material information

Constituent 1

Test material form:

other: Cream powder (84% w/w)

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL (MI3 dried)
- Source and lot/batch No.of test material: C202483
- Expiration date of the lot/batch: 2001.05.31
- Purity test date: 2001.05.31

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: < -10 degrees Celcius
- Stability under test conditions: NA
- Final preparation of a solid: dosed as powder/soild at 2000mg/kg bw

FORM AS APPLIED IN THE TEST (if different from that of starting material): as solid/ powder

Test animals

Species:

rat

Strain:

Wistar

Remarks:

CRL:Han Wist(Glx/BRL)BL)

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Crl:WI(Glx/BRL/Han)BR)
- Females (if applicable) nulliparous and non-pregnant: yes
- Age at study initiation: 7 - 8 weeks
- Weight at study initiation: 188 -208g (males); 137 - 163g (females)
- Fasting period before study: yes, overnight fasting.
- Housing: Three rats of same sex per cage (suspended stainless steel mesh cage).
- Diet (e.g. ad libitum): ad libitum.
- Water (e.g. ad libitum): ad libitum.
- Acclimation period: 22- 24 days.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19 - 25 degrees Celcuis.
- Humidity (%): 30 - 70%.
- Air changes (per hr): a minimum of 14 air changes per hour.
- Photoperiod (hrs dark / hrs light): 12- / 12 hours

IN-LIFE DATES: 31- 38 days.

Administration / exposure

Type of coverage:

semiocclusive

Vehicle:

unchanged (no vehicle)

Remarks:

Wetting of the application area with 0.2 mL water before adding MI3 powder

Details on dermal exposure:

TEST SITE
- Area of exposure: 5 x 5 cm
- % coverage: 10% of the total body surface
- Type of wrap if used: Semi occlusive

REMOVAL OF TEST SUBSTANCE
- Washing (if done): brushed and swabbed with moist cotton wool
- Time after start of exposure: 24 hours

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 2000 mg/kg bw
- Concentration (if solution): NA
- Constant volume or concentration used: NA
- For solids, paste formed: no

VEHICLE
Not used, application area moistened with water before application of powder.

Duration of exposure:

24 hours.

Doses:

single dose of 2000 mg/kg bw.

No. of animals per sex per dose:

2 female rats (Preliminary study) and 5 males and 5 females (main study)

Control animals:

no

Details on study design:

Duration of observation period following administration: 8 days (preliminary study) 14 days (main study)
- Frequency of observations and weighing:
Clinical signs:The day of dosing (fist day), once within half an hour of dosing and four times within the first four hours after administration. Twice daily on Days 2, 3 and 4, and once daily from the fifth to the last day of the observation period. Weighing: one day before dosing (day -1), day 1, 8 and 15.
Dermal reactions were recorded from day 2 of exposure and similar to that for clinical signs.
- Necropsy of survivors performed: yes
- Other examinations performed: clinical signs, body weight,

Statistics:

NA

Results and discussion

Preliminary study:

Yes: Two female rats dosed at 2000 mg/kg bw. Based on the results these investigations the limie dose level was selected for the main study. Since no compound-related motality occured in the preliminary test, a group of five male and female rats was subjected to a single dermal application of MI3 at 2000 mg/kg bw.

Effect levels

Key result

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat. (total fraction)

Remarks on result:

no indication of skin irritation up to the relevant limit dose level

Mortality:

No mortality was observed neither in the preliminary study nor the main study.

Clinical signs:

- No signs in the preliminary study.
- Three incidence of anogenital soiling for 3 female rats at day 2, recovery at day 3. No dermal reactions both sex.

Body weight:

all rats gained weight during the first and second week.

Gross pathology:

One male and female rat with slight pelvic dilatation of one kidney.

Other findings:

NA

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

The Study was conducted to access the acute dermal toxicity of MI3 given as single dose to a small group of rats of both sex. The study was designed in accordance with OECD guideline method 402. In a preliminary study two female rats and in the main study groups of five male and female rats were given a single dose of MI3 at a dose level of 2000 mg/kg bw. All animals were killed at day 8 (preliminary study) or day 15 (main study) after dosing. No death were observed, in the main study transient anogenital soiling was observed among three female rats, and one male and one female rat had slight pelvic dilatation of one kidney. The median LD50 of MI3 in rats was found to be > 2000 mg/kg bw.

Executive summary:

The Study was conducted to access the acute dermal toxicity of MI3 given as single dose to a small group of rats of both sex. The study was designed in accordance with OECD guideline method 402, the commission directive 92/69/EEC, Method B3, and the US EPA Health effects test Guidelines OPPTS 870.1200.

In the preliminary study two female rats had undiluted MI3 applied to the dorsa at a dose level of 2000 mg/kg bw. In the main study, groups of five male and female rats were given a single topical application of MI3 at a dose level of 2000 mg/kg bw to the dorsum with a semi occlusive bandage covering the dose site for 24 hours. Animals of the preliminary study were killed at day 8, while animals of the main study were killed at day 15. All animals underwent a full necropsy.

Weight gain was recorded among all animals during the study period. In the preliminary study, no death or macroscopic changes were evident 8 days after dosing. In the main study, transient anogenital soiling was observed among three female rats on day 2 with recovery on day 3. On day 15 one male and one female rat had slight pelvic dilatation of one kidney.

The median LD50 of MI3 in rats was found to be > 2000 mg/kg bw.