Insulin Aspart Ethyl Ester

Acute Toxicity: oral

Administrative data

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2000.03.31 - 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:

2000

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

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

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

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

Deviations:

no

Test guideline 3

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.1100 (Acute Oral Toxicity)

Deviations:

no

Principles of method if other than guideline:

NA

GLP compliance:

yes (incl. QA statement)

Test type:

acute toxic class method

Limit test:

yes

Test material

Test material information

Constituent 1

Reference substance name:

X14DesB30

Molecular formula:

X14DesB30

IUPAC Name:

X14DesB30

Test material form:

solid: particulate/powder

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL (X14DesB30, solid)
- Source and lot/batch No.of test material: X1YIS0201014
- Expiration date of the lot/batch: NA
- Purity test date: 2000.02.18

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: < -70 degrees Celcius
- Stability under test conditions: NA
- Solubility and stability of the test substance in the solvent/vehicle: Full solubility in vehicle (water)
- Reactivity of the test substance with the solvent/vehicle of the cell culture medium: NA

TREATMENT OF TEST MATERIAL PRIOR TO TESTING
- Treatment of test material prior to testing: Purified water (Elgastat purifier used)
- Final preparation of a solid: 100mg/ml X14DesB30 in water

FORM AS APPLIED IN THE TEST (if different from that of starting material): solid disolved in purified water.

Test animals

Species:

rat

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: 10 - 11 weeks
- Weight at study initiation: 278 – 336g (males); 180 - 194g (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: 14 - 16 days.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19 - 25 degrees Celcuis.
- Humidity (%): 40 - 70%.
- Air changes (per hr): a minimum of 14 air changes per hour.
- Photoperiod (hrs dark / hrs light): 12- / 12 hours

IN-LIFE DATES: From: To: 29 - 31 days.

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

water

Details on oral exposure:

VEHICLE
- Concentration in vehicle: 100 mg/ml
- Amount of vehicle (if gavage): 20 ml/kg
- Justification for choice of vehicle: MI3 was dispersed in purified water, which is non-toxic in the volume given.
- Lot/batch no. (if required): C202483
- Purity: Pure water (Elgastat purifier)

MAXIMUM DOSE VOLUME APPLIED: 20 ml/kg bw

DOSAGE PREPARATION (if unusual): NA

CLASS METHOD (if applicable)
- Rationale for the selection of the starting dose: Dose levels were chosen in complience with limits stated in the guidelines - i.e. first dose 2000 mg/kg to three animals of one sex.

Doses:

2000 mg/kg bw

No. of animals per sex per dose:

3 (females) / 3 (males)

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing:
Clinical signs: The day of dosing (fist day), one 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.
- Necropsy of survivors performed: yes
- Other examinations performed: clinical signs, body weight,

Statistics:

NA

Results and discussion

Effect levels

Key result

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat. (total fraction)

Mortality:

There were no deaths following a single oral dose of X14desB30 of both sexes dosed at 2000 mg/kg.

Clinical signs:

There were no clinical signs of reaction to treatment were apparent apart from a single transient observation of pilo-erection for one female one hour after dosing.

Body weight:

Two male and three females showed low weight gains or small losses between Day -1 and Day 8. The females continued to show small gains during the second week of the study.
The majority of rats gained weight during the second week of their observation period.

Gross pathology:

No macroscopic changes were observed for animals killed on Day 15.

Other findings:

NA

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

The Study was conducted to access the acute oral toxicity of X14DesB30 given as single dose to a small group of rats of both sex. The study was designed in accordance with OECD guideline method 423. Groups of three male or female rats were fasted overnight and given a single dose of X14DesB30 by oral gavage at a dose level of 2000 mg/kg. All animals were killed at day 15 after dosing. No death or post mortem abnormalities were observed for rats of both sexes after a single oral dose of 2000 mg/kg. However, a single transient observation of pilo-erection was observed for one female one hour after dosing. The median lethal dose 50 of X14DesB30 in rats was found to be > 2000 mg/kg bw.

Executive summary:

The Study was conducted to access the acute oral toxicity of X14DesB30 given as single dose to a small group of rats of both sex. The study was designed in accordance with OECD guideline method 423, the commission directive 94/54/EEC, Method B1 tris, and the US EPA Health effects test Guidelines OPPTS 870.1100.

The study design provided information for hazard assessment and classification and enables the X14DesB30 to be assigned to one of the four toxicity classes identified in Annex of Commission Directive 93/21/EEC, while severely restricting animal usage.

Groups of three male and female rats were fasted overnight and given a single dose of MI3 oral gavage at a dose level of 2000 mg/kg. The X14DesB30 was dispersed in purified water and dosed at a dose volume of 20 mL/kg. All animals were killed at day 15 after dosing and underwent a full necropsy.

No deaths were observed for rats of both sexes after a single oral dose of 2000 mg/kg. No clinical signs were observed, apart from a single transient observation of pilo-erection for one female one hour after dosing. Two male and three females showed low weight gains or small losses the first week. The females continued to show small gains during the second week of the study, although the majority of rats gained weight during the second week of their observation period. Necropsy did not reveal any macroscopic abnormalities among the dosed animals.

The median lethal dose 50 of X14DesB30 in rats was found to be > 2000 mg/kg bw.