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Toxicological information

Skin sensitisation

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Administrative data

Endpoint:
skin sensitisation: in vitro
Type of information:
experimental study
Adequacy of study:
key study
Study period:
14 August 2017 to 31 October 2017 and 7 November 2017 to 14 March 2018
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Referenceopen allclose all

Reference Type:
study report
Title:
Unnamed
Year:
2017
Report Date:
2017
Reference Type:
study report
Title:
Unnamed
Year:
2018
Report Date:
2018

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 442D (In Vitro Skin Sensitisation: ARE-Nrf2 Luciferase Test Method)
Version / remarks:
OECD Guideline TG 442D: In Vitro Skin Sensitisation: ARE-Nrf2 Luciferase Test Method (adopted February, 2015).
Deviations:
no
GLP compliance:
yes
Type of study:
activation of keratinocytes

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
liquid
Details on test material:
CAS 1629579-82-3
Molecular formula C18H32N2O11
EC 818-033-1

Results and discussion

Positive control results:
The luciferase activity induction obtained with the positive control, Ethylene dimethacrylate glycol, was above the threshold of 1.5-fold in at least one concentration.
The EC1.5 of the positive control was between 5 and 125 µM (43 µM and 63 µM in experiment 1 and 2, respectively). A dose response was observed and the induction at 250 µM was higher than 2-fold (2.57-fold and 2.35-fold in experiment 1 and 2, respectively).

In vitro / in chemico

Resultsopen allclose all
Key result
Parameter:
other: Maximum luciferase activity (Imax fold)
Run / experiment:
1
Value:
ca. 1.76
Vehicle controls valid:
yes
Negative controls valid:
yes
Positive controls valid:
yes
Key result
Parameter:
other: Maximum luciferase activity (Imax fold)
Run / experiment:
2
Value:
ca. 1.51
Vehicle controls valid:
yes
Negative controls valid:
yes
Positive controls valid:
yes
Key result
Parameter:
other: Maximum luciferase activity
Remarks:
(lmax fold)
Run / experiment:
3 (2018)
Value:
ca. 2.36
Vehicle controls valid:
yes
Negative controls valid:
yes
Positive controls valid:
yes

Any other information on results incl. tables

The average coefficient of variation of the luminescence reading for the negative (solvent) control DMSO was below 20% (4.8% and 5.9% in experiment 1 and 2, respectively).

Applicant's summary and conclusion

Conclusions:
2018 study
In conclusion, based on the current study and the two experiments performed in Test Facility Study No. 518597, Bis-Aminopropyl Diglycol Dimaleate is classified as positive (activation of the antioxidant/electrophile responsive element (ARE)-dependent pathway in keratinocytes) in the KeratinoSensTM assay

2017 study
The test item showed no toxicity (no IC30 and IC50 value) and no biologically relevant induction of the luciferase activity (EC1.5 values of 1048 µM and 1975 µM in experiment 1 and 2, respectively) was measured at concentrations lower than 1000 µM in both experiments. The maximum luciferase activity induction (Imax) was 1.76-fold and 1.51-fold in experiment 1 and 2 respectively. The test item is classified as negative in the KeratinoSensTM assay since in both experiments no biological relevant luminescence induction at concentrations below 1000 µM was observed.
In conclusion, Bis-Aminopropyl Diglycol Dimaleate is classified as negative (no biological relevant activation of the antioxidant/electrophile responsive element (ARE)-dependent pathway in keratinocytes) under the experimental conditions described in this report.
Executive summary:

2018 study

The objective of this study was to evaluate the ability of Bis-Aminopropyl Diglycol Dimaleateto activate theantioxidant/electrophile responsive element (ARE)-dependent pathway in the KeratinoSensÔassay.

In Test Facility Study No. 518597 two KeratinoSens experiments were performed. One assay run gave a positive result (EC1.5of 757 µM) and the second assay run gave a negative result (EC1.5of 1427 but no induction below 1000 µM). At first instance the conclusion of Test Facility Study No. 518597 was negative but due to a change in the correction factor used for the correction of the water content no conclusion about the classification was possible since the results of the two experiments in Test Facility Study No. 518597 were not concordant. A third assay run (experiment) was required for a classification in the KeratinoSensTMassay. This assay run is performed in the current study.

The study procedures described in this report were based on the most recent OECD guideline.

Batch 11_13_17_1 of Bis-Aminopropyl Diglycol Dimaleate was a clear to transparent yellow liquid. A correction factor of 3.85 was used to correct for the water content (74%). Bis-Aminopropyl Diglycol Dimaleate was dissolved in dimethyl sulfoxide at 76 mM. From this stock 11 spike solutions in DMSO were prepared. The stock and spike solutions were diluted 100-fold in the assay resulting in test concentrations of0.38 – 776 µM (2-fold dilution series). 

No precipitate was observed at any dose level tested. One experiment was performed.

The experiment passed the acceptance criteria:

·        The luciferase activity induction obtained with the positive control,Ethylene dimethacrylate glycol, was above the threshold of 1.5-fold in at least one concentration. 

·        The EC1.5of the positive control was between 5 and 125 µM (96 µM). A dose response was observed and the induction at 250 µM was higher than 2-fold (3.17-fold).

·        Finally, the average coefficient of variation of the luminescence reading for the negative (solvent) control DMSO was below 20% (6.6%).

Overall it is concluded that the test conditions were adequate and that the test system functioned properly. 

Bis-Aminopropyl Diglycol Dimaleate showed no toxicity (no IC30and IC50value) in the current experiment. This result is similar to the results that were observed in the two experiments performed inTest Facility Study No.518597.

A biologically relevant, dose-related induction of the luciferase activity (EC1.5value 269 µM) with a maximum induction of 2.36-fold was measured in the current experiment. In Test Facility Study No. 518597 two KeratinoSens runs were performed. One assay run gave a positive result (EC1.5of 757 µM) and the second assay run gave a negative result (EC1.5of 1427 µM but no induction below 1000 µM).

Bis-Aminopropyl Diglycol Dimaleateis classified as positive in the KeratinoSensTMassay sincepositive results (>1.5-fold induction) were observed in 2 out of 3 experiments at test concentrations ≤ 1000µMwith a cell viability >70%.

In conclusion, based on the current study and the two experiments performed in Test Facility Study No. 518597, Bis-Aminopropyl Diglycol Dimaleate is classified as positive(activation of theantioxidant/electrophile responsive element (ARE)-dependent pathway in keratinocytes) in the KeratinoSensTMassay

2017 study

The objective of this study was to evaluate the ability of Bis-Aminopropyl Diglycol Dimaleate to activate the antioxidant/electrophile responsive element (ARE)-dependent pathway in the KeratinoSensTM assay.

The study procedures described in this report were based on the most recent OECD guideline.

Batch 3 of the test item was a clear colourless to pale yellow liquid. A correction factor of 2.778 was used to correct for the water content (74% water). The test item was dissolved in dimethyl sulfoxide at 200 mM. From this stock 11 spike solutions in DMSO were prepared. The stock and spike solutions were diluted 100-fold in the assay resulting in test concentrations of 0.98 – 2000 µM (2-fold dilution steps) in experiment 1 and of 172 – 2000 µM (1.25-fold) in experiment 2. In the second experiment, a more narrow dose-response analysis was performed using a lower dilution factor of 1.25-fold to investigate the induction at 1000 µM in experiment 1 in more detail. Two independent experiments were performed.

Both experiments passed the acceptance criteria:

  • The luciferase activity induction obtained with the positive control, Ethylene dimethacrylate glycol, was above the threshold of 1.5-fold in at least one concentration.
  • The EC1.5 of the positive control was between 5 and 125 µM (43 µM and 63 µM in experiment 1 and 2, respectively). A dose response was observed and the induction at 250 µM was higher than 2-fold (2.57 -fold and 2.35-fold in experiment 1 and 2, respectively).
  • Finally, the average coefficient of variation of the luminescence reading for the negative (solvent) control DMSO was below 20% (4.8% and 5.9% in experiment 1 and 2, respectively).

Overall it is concluded that the test conditions were adequate and that the test system functioned properly.

The test item showed no toxicity (no IC30 and IC50 value) and no biologically relevant induction of the luciferase activity (EC1.5 values of 1048 µM and 1975 µM in experiment 1 and 2, respectively) was measured at concentrations lower than 1000 µM in both experiments. The maximum luciferase activity induction (Imax) was 1.76-fold and 1.51-fold in experiment 1 and 2 respectively. The test item is classified as negative in the KeratinoSensTM assay since in both experiments no biological relevant luminescence induction at concentrations below 1000 µM was observed.

In conclusion, Bis-Aminopropyl Diglycol Dimaleate is classified as negative (no biological relevant activation of the antioxidant/electrophile responsive element (ARE)-dependent pathway in keratinocytes) under the experimental conditions described in this report.