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Please be aware that this old REACH registration data factsheet is no longer maintained; it remains frozen as of 19th May 2023.

The new ECHA CHEM database has been released by ECHA, and it now contains all REACH registration data. There are more details on the transition of ECHA's published data to ECHA CHEM here.

Diss Factsheets

Administrative data

Endpoint:
in vitro gene mutation study in bacteria
Remarks:
Type of genotoxicity: gene mutation
Type of information:
experimental study
Adequacy of study:
key study
Study period:
From 07 Mar to 29 Mar 2016
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Study run to a method comparable with current guidelines and to GLP

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2016
Report date:
2016

Materials and methods

Test guideline
Qualifier:
according to guideline
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
Deviations:
no
GLP compliance:
yes
Type of assay:
bacterial reverse mutation assay

Test material

Constituent 1
Chemical structure
Reference substance name:
2-methylbutan-2-ol; tert-butyl N-{5-bromo-3-[(1R)-1-(2-{[(5-cyano-1-methyl-1H-pyrazol-3-yl)methyl](methyl)carbamoyl}-5-fluorophenyl)ethoxy]pyridin-2-yl}-N-[(tert-butoxy)carbonyl]carbamate
EC Number:
812-621-1
Cas Number:
1910113-99-3
Molecular formula:
C31H36BrFN6O6•C5H12O
IUPAC Name:
2-methylbutan-2-ol; tert-butyl N-{5-bromo-3-[(1R)-1-(2-{[(5-cyano-1-methyl-1H-pyrazol-3-yl)methyl](methyl)carbamoyl}-5-fluorophenyl)ethoxy]pyridin-2-yl}-N-[(tert-butoxy)carbonyl]carbamate
Constituent 2
Reference substance name:
di-tert-butyl {5-bromo-3-[(1R)-1-(2-{[(5-cyano-1-methyl-1H-pyrazol-3-yl)methyl](methyl)carbamoyl}-5- fluorophenyl)ethoxy]pyridin-2-yl}imidodicarbonate t-amyl alcohol solvate
IUPAC Name:
di-tert-butyl {5-bromo-3-[(1R)-1-(2-{[(5-cyano-1-methyl-1H-pyrazol-3-yl)methyl](methyl)carbamoyl}-5- fluorophenyl)ethoxy]pyridin-2-yl}imidodicarbonate t-amyl alcohol solvate
Test material form:
solid: particulate/powder
Remarks:
migrated information: powder
Details on test material:
Batch No.: E010016713
Purity: not specified

Method

Target gene:
Histidine gene in Salmonella typhimurium, tryptophan gene in Escherichia coli
Species / strainopen allclose all
Species / strain / cell type:
S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
Species / strain / cell type:
E. coli WP2 uvr A
Metabolic activation:
with and without
Metabolic activation system:
S9 mix
Test concentrations with justification for top dose:
Dose range finding test/first mutation experiment: 1.7, 5.4, 17, 52, 164, 512, 1600 and 5000 μg/plate
Experiment 2: 154, 275, 492, 1180 and 2800 μg/plate
Vehicle / solvent:
- Vehicle(s)/solvent(s) used: dimethyl sulfoxide
- Justification for choice of solvent/vehicle: A solubility test was performed and the test item was dissolved in dimethyl sulfoxide.
Controlsopen allclose all
Untreated negative controls:
no
Negative solvent / vehicle controls:
yes
True negative controls:
no
Positive controls:
yes
Positive control substance:
sodium azide
Remarks:
In the absence of S-9 mix for TA1535 strains
Untreated negative controls:
no
Negative solvent / vehicle controls:
yes
True negative controls:
no
Positive controls:
yes
Positive control substance:
other: ICR-191
Remarks:
In the absence of S-9 mix for TA1537 strains.
Untreated negative controls:
no
Negative solvent / vehicle controls:
yes
True negative controls:
no
Positive controls:
yes
Positive control substance:
2-nitrofluorene
Remarks:
In the absence of S-9 mix for TA98 strains.
Untreated negative controls:
no
Negative solvent / vehicle controls:
yes
True negative controls:
no
Positive controls:
yes
Positive control substance:
methylmethanesulfonate
Remarks:
In the absence of S-9 mix for TA100 strains.
Untreated negative controls:
no
Negative solvent / vehicle controls:
yes
True negative controls:
no
Positive controls:
yes
Positive control substance:
4-nitroquinoline-N-oxide
Remarks:
In the absence of S-9 mix for WP2uvrA strains.
Untreated negative controls:
no
Negative solvent / vehicle controls:
yes
True negative controls:
no
Positive controls:
yes
Positive control substance:
other: 2-aminoanthracene
Remarks:
In the presence of S-9 mix for TA 1535, TA 1537, TA 98, TA 100 and WP2uvrA strains.
Details on test system and experimental conditions:
METHOD OF APPLICATION: Dose range finding test and experiment 2: in agar (plate incorporation)
DURATION
- Preincubation period:
- Exposure duration: 48 ± 4 h both for dose range finding test and experiment 2
- Expression time (cells in growth medium):
- Selection time (if incubation with a selection agent):
- Fixation time (start of exposure up to fixation or harvest of cells):

SELECTION AGENT (mutation assays):
SPINDLE INHIBITOR (cytogenetic assays):
STAIN (for cytogenetic assays):

NUMBER OF REPLICATIONS: 3 replications both for dose range finding test and experiment 2

NUMBER OF CELLS EVALUATED:

DETERMINATION OF CYTOTOXICITY
- Method: cloning efficiency

OTHER EXAMINATIONS:
- Determination of polyploidy:
- Determination of endoreplication:
- Other: Evidence of test item precipitate on the plates and the condition of the bacterial background lawn were evaluated when considered necessary, macroscopically and/or microscopically by using a dissecting microscope.

OTHER:
Evaluation criteria:
A Salmonella typhimurium reverse mutation assay and/or Escherichia coli reverse mutation assay is considered acceptable if it meets the following criteria:
a) The vehicle control and positive control plates from each tester strain (with or without S9-mix) must exhibit a characteristic number of revertant colonies when compared against relevant historical control data generated at WIL Research Europe.
b) The selected dose range should include a clearly toxic concentration or should exhibit limited solubility as demonstrated by the preliminary toxicity range-finding test or should extend to 5 mg/plate.
c) No more than 5% of the plates are lost through contamination or some other unforeseen event. If the results are considered invalid due to contamination, the experiment will be repeated.
Statistics:
A test item is considered negative (not mutagenic) in the test if:
a) The total number of revertants in the tester strain TA100 or WP2uvrA is not greater than two (2) times the concurrent control, and the total number of revertants in tester strains TA1535, TA1537 or TA98 is not greater than three (3) times the concurrent vehicle control.
b) The negative response should be reproducible in at least one follow-up experiment.
A test item is considered positive (mutagenic) in the test if:
a) The total number of revertants in the tester strain TA100 or WP2uvrA is greater than two (2) times the concurrent control, or the total number of revertants in tester strains TA1535, TA1537, TA98 is greater than three (3) times the concurrent vehicle control.
b) In case a follow up experiment is performed when a positive response is observed in one of the tester strains, the positive response should be reproducible in at least one follow up experiment.

Results and discussion

Test resultsopen allclose all
Species / strain:
S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
not specified
Vehicle controls validity:
valid
Positive controls validity:
valid
Species / strain:
E. coli WP2 uvr A
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
not specified
Vehicle controls validity:
valid
Positive controls validity:
valid
Additional information on results:
Dose range finding test/first mutation experiment:
- Precipitate: Precipitation of test substance on the plates was observed at the start and at the end of the incubation period at concentrations of 1600 and 5000 μg/plate, except in the first mutation experiment, where precipitation already was observed at the concentration of 512 μg/plate at the start of the incubation period.
- Toxicity: To determine the toxicity of the test substance, the reduction of the bacterial background lawn, the increase in the size of the microcolonies and the reduction of the revertant colonies were examined. No reduction of the bacterial background lawn revertants was observed. No biologically relevant decrease in the number of revertants was observed up to the dose level of 5000 μg/plate (dose range finding) and 1600 μg/plate (first mutation experiment). Since the test substance precipitated heavily on the plates at the test substance concentration of 5000 μg/plate in the first mutation experiment, the number of revertants of this dose level could not be determined.
- Mutagenicity: No increase in the number of revertants was observed upon treatment with the test item under all conditions tested.

Experiment 2:
- Precipitate: Precipitation of test substance on the plates was observed at the start and at the end of the incubation period at concentrations of 1180 and 2800 μg/plate.
- Toxicity: In the second mutation assay, there was no reduction of the bacterial background lawn. No biologically relevant decrease in the number of revertants was observed up to the dose level of 1180 μg/plate. Since the test substance precipitated heavily on the plates at the test substance concentration of 2800 μg/plate, the number of revertants of this dose level could not be determined.
- Mutagenicity: In the second mutation assay, no increase in the number of revertants was observed upon treatment with test substance under all conditions tested.
Remarks on result:
other: all strains/cell types tested
Remarks:
Migrated from field 'Test system'.

Applicant's summary and conclusion

Conclusions:
Interpretation of results (migrated information):
negative

Based on the results of this study it is concluded that the test substance is not mutagenic in the Salmonella typhimurium reverse mutation assay and in the Escherichia coli reverse mutation assay.