Registration Dossier

Diss Factsheets

Administrative data

Endpoint:
skin irritation: in vitro / ex vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
31 October 2016 - 30 Nov 2016
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2017
Report date:
2017

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to guideline
Guideline:
OECD Guideline 439 (In Vitro Skin Irritation: Reconstructed Human Epidermis Test Method)
Deviations:
no
Qualifier:
according to guideline
Guideline:
EU Method B.46 (In Vitro Skin Irritation: Reconstructed Human Epidermis Model Test)
Deviations:
no
GLP compliance:
yes
Remarks:
including compliance statement and signature page

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
solid: bulk

In vitro test system

Test system:
human skin model
Remarks:
EpiDermTM SIT (EPI-200) three-dimensional human skin model
Cell type:
non-transformed keratinocytes
Vehicle:
other: deionised water and isopropanol
Details on test system:
EpiDermTM SIT (EPI-200) tissues were kept in their packaging until the next step. The tissues
were set up the day prior to treatment by placing each tissue onto 0.9 mL maintenance
medium (supplied with the EpiDermTM SIT (EPI-200) tissues) in 6-well plates and incubating
at 37°C.
Control samples:
yes, concurrent negative control
yes, concurrent positive control
yes, concurrent MTT non-specific colour control
Amount/concentration applied:
25 mg of test article was added to 1 mL of MTT In order to assess the potential non-specific reduction of the test article.
25 mg of the test article was added to 0.3 mL deionised water and 0.3 mL isopropanol in a suitable glass container in order to assess the potential of staining.
25 mg were used for exam of test substance.

A volume of 30 μL of the positive and negative control solutions was used.
Duration of treatment / exposure:
The treated tissues were paced into an incubator at 37±1ºC, 5±1% CO2 for 35 minutes. The
plates were removed from the incubator and placed into a sterile hood until the 60 minute
treatment period was complete for each tissue. Following treatment, substances were
removed by washing the tissues.
Duration of post-treatment incubation (if applicable):
The tissues were then placed on the appropriate medium and
incubated for 41 hours and 20 minutes.
Number of replicates:
3

Results and discussion

In vitro

Results
Irritation / corrosion parameter:
% tissue viability
Value:
ca. 44.6
Negative controls validity:
valid
Remarks:
100.0%
Positive controls validity:
valid
Remarks:
4.4%
Remarks on result:
positive indication of irritation

Any other information on results incl. tables

The test article, IN 79066, was considered to be irritant in the in vitro skin model EpiDermTM

SIT (EPI-200).

Applicant's summary and conclusion

Interpretation of results:
Category 1 (corrosive) based on GHS criteria
Conclusions:
This study was conducted to determine whether the test article causes irritation in the in vitro
skin model EpiDermTM SIT (EPI-200).
EpiDermTM SIT (EPI-200) inserts were treated with IN 79066, negative control (phosphate
buffered saline (PBS)) and positive control (5% w/v sodium dodecyl sulphate (SDS)) for
60 minutes. At the end of the treatment period, the tissues were washed with PBS and cell
viability was assessed using the MTT assay. The skin irritation potential was classified
according to the remaining cell viability obtained after test article treatment.
The group mean viability for the test article was 44.6%, for the negative control was 100%
and for the positive control was 4.4%.
The test article, IN 79066, was considered to be irritant in the in vitro skin model EpiDermTM
SIT (EPI-200).
Corrosive properties of BI 730357 Amin My can’t be excluded as OECD 431 test is not available. Therefore Category 1 is classified.