Insulin Aspart Precursor

Skin irritation / corrosion

Administrative data

Endpoint:

skin irritation: in vivo

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 404 (Acute Dermal Irritation / Corrosion)

Deviations:

yes

Remarks:

The relative humidity in the animal holding room was consistently lower (15 %) than stated in the protocol. The sentinel rabbit assessments were conducted on Day 9 rather than Day 8. The deviations are not considered to compromise the study integrity

Test guideline 2

Qualifier:

according to guideline

Guideline:

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

Deviations:

yes

Remarks:

The relative humidity in the animal holding room was consistently lower (15 %) than stated in the protocol. The sentinel rabbit assessments were conducted on Day 9 rather than Day 8. The deviations are not considered to compromise the study integrity

Test guideline 3

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.2500 (Acute Dermal Irritation)

Deviations:

yes

Remarks:

The relative humidity in the animal holding room was consistently lower (15 %) than stated in the protocol. The sentinel rabbit assessments were conducted on Day 9 rather than Day 8. The deviations are not considered to compromise the study integrity

GLP compliance:

yes (incl. QA statement)

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)
- Source and lot/batch No.of test material: X1YIS0201014
- Expiration date of the lot/batch: NA
- Purity 80%
- 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: Soluble 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.
- Final preparation of a solid: A 50% m/v dispersion in purified water, pH of 3.22.

FORM AS APPLIED IN THE TEST (if different from that of starting material): A 50% m/v dispersion in purified water.

Test animals

Species:

rabbit

Strain:

New Zealand White

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Rabbit (New Zealand White rabbit)
- Sex: male and female
- Age at study initiation: 11 - 13 weeks
- Weight at study initiation: 2.35 - 2.41 kg
- Housing: Single housed in floor-pens with wood chips bedding
- Diet (e.g. ad libitum): ad libitum
- Water (e.g. ad libitum): ad libitum
- Acclimation period: NA

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 16-22
- Humidity (%): 40 – 80 %
- Air changes (per hr): at least 10 air changes per hour.
- Photoperiod (hrs dark / hrs light): 12 hours/12 hours

IN-LIFE DATES: From: To: at least 17 days

Test system

Type of coverage:

semiocclusive

Preparation of test site:

clipped

Vehicle:

water

Remarks:

50% m/v dispersion, pH of 3.22

Controls:

no

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.5mL X14DesB30 as a paste

Duration of treatment / exposure:

four hours

Observation period:

One hour, 24, 48 and 72 hours

Number of animals:

3 (one male and two females)

Details on study design:

TEST SITE
- Area of exposure: 30 x 20 mm
- % coverage: NA
- Type of wrap if used: Dense elastic gauze bandage (semi-occlusive)

REMOVAL OF TEST SUBSTANCE
- Washing (if done): Light brushing and swabbing with moistened cotton wool
- Time after start of exposure: 4 hours

OBSERVATION TIME POINTS
(indicate if minutes, hours or days) one hour, 24, 48 and 72 hours after application.

Results and discussion

In vivo

Results

Results 1

Irritation parameter:

erythema score

Basis:

animal #1

Time point:

other: 1/24/48/72 h

Score:

0

Max. score:

0

Remarks on result:

other: Petechial haemorrhage (transient - recovery after 4 days) present after removal of dose residuals.

Results 2

Irritation parameter:

erythema score

Basis:

animal #2

Time point:

other: 1/24/48/72 h

Score:

0

Max. score:

0

Remarks on result:

other: Petechial hemorrhage (transient - recovery after 24h) present after removal of dose residuals. Developed scabs by Day 3 and showed full resolution by Day 4.

Results 3

Irritation parameter:

erythema score

Basis:

animal #3

Time point:

other: 1/24/48/72 h

Score:

0

Max. score:

0

Remarks on result:

other: Petechial hemorrhage (transient - recovery after 24h) present after removal of dose residuals. Developed scabs by Day 3 and showed full resolution by Day 4.

Results 4

Irritation parameter:

edema score

Basis:

animal #1

Time point:

other: 1/24/48/72 h

Score:

0

Max. score:

0

Remarks on result:

other: Petechial haemorrhage (transient - recovery after 4 days) present after removal of dose residuals.

Results 5

Irritation parameter:

edema score

Basis:

animal #2

Time point:

other: 1/24/48/72 h

Score:

0

Max. score:

0

Remarks on result:

other: Petechial hemorrhage (transient - recovery after 24h) present after removal of dose residuals. Developed scabs by Day 3 and showed full resolution by Day 4.

Results 6

Irritation parameter:

edema score

Basis:

animal #3

Time point:

other: 1/24/48/72 h

Score:

0

Max. score:

0

Remarks on result:

other: Petechial hemorrhage (transient - recovery after 24h) present after removal of dose residuals. Developed scabs by Day 3 and showed full resolution by Day 4.

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

The Study was conducted to determine the irritation caused to intact rabbit skin following a single (4 hour), semi-occluded, topical application of X14DesB30. The study was designed in accordance with OECD guideline method 404. One male and two female rabbits were exposed to a single dose of 0.5 mL X14DesB30 applied to a 30 x 20 mm area on the clipped dorsum. All animals were killed by an intravenous injection of an overdose of barbiturate at day 4. X14DesB30 did not cause any notable dermal reactions. All observed reactions were a result of mechanical trauma caused by removal of the dose residuals.

Executive summary:

The Study was conducted to determine the irritation caused to intact rabbit skin following a single (4 hour), semi-occluded, topical application of X14DesB30. The study was designed in accordance with OECD guideline method 404, the commission directive 92/69/EEC, Method B4, and the US EPA Health effects test Guidelines OPPTS 870.2500. A single dose of 0.5 mL X14DesB30 was applied to a 30 x 20 mm area on the clipped dorsum of each of three rabbits, one male and two female rabbits. A semi- occlusive bandage covered the treated area of skin for 4 hours and dermal reactions to treatment were assessed for up to nine days after removal of the dressings. All animals were killed by an intravenous injection of an overdose of barbiturate at Day four or Day nine. Transient petechial hemorrhage was observed as a result of mechanical trauma caused by removal of the dose residuals. Summarized, X14DesB30 did not cause any notable dermal reactions and classified not to cause skin irritation.