Reaction mass of rel-{(1R,2S)-1-methyl-2-[(2R)-5-methylhex-4-en-2-yl]cyclopropyl}methanol and rel-{(1S,2R)-1-methyl-2-[(2R)-5-methylhex-4-en-2-yl]cyclopropyl}methanol

Genetic toxicity: in vitro

Administrative data

Endpoint:

in vitro cytogenicity / chromosome aberration study in mammalian cells

Remarks:

Type of genotoxicity: chromosome aberration

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2015

Reliability:

1 (reliable without restriction)

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Report date:

2015

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 473 (In Vitro Mammalian Chromosome Aberration Test)

GLP compliance:

yes (incl. QA statement)

Type of assay:

in vitro mammalian chromosome aberration test

Test material

Test material information

Constituent 1

Reference substance name:

[(1R,2S)-1-methyl-2-[(2R)-5-methylhex-4-en-2-yl]cyclopropyl]methanol; [(1R,2S)-1-methyl-2-[(2S)-5-methylhex-4-en-2-yl]cyclopropyl]methanol; [(1S,2R)-1-methyl-2-[(2R)-5-methylhex-4-en-2-yl]cyclopropyl]methanol; [(1S,2R)-1-methyl-2-[(2S)-5-methylhex-4-en-2-yl]cyclopropyl]methanol

EC Number:

942-597-9

Cas Number:

1655500-83-6

Molecular formula:

C12H22O

IUPAC Name:

[(1R,2S)-1-methyl-2-[(2R)-5-methylhex-4-en-2-yl]cyclopropyl]methanol; [(1R,2S)-1-methyl-2-[(2S)-5-methylhex-4-en-2-yl]cyclopropyl]methanol; [(1S,2R)-1-methyl-2-[(2R)-5-methylhex-4-en-2-yl]cyclopropyl]methanol; [(1S,2R)-1-methyl-2-[(2S)-5-methylhex-4-en-2-yl]cyclopropyl]methanol

Test material form:

other: liquid

Details on test material:

Batch VE00340479
Purity 87.8%

Specific details on test material used for the study:

GR-50-1408 is the Givaudan identification code which was employed for ROSYFOLIA during the early, developmental and testing period.

Method

Species / strain

Species / strain / cell type:

lymphocytes: Human

Metabolic activation:

with and without

Metabolic activation system:

S9 mix

Test concentrations with justification for top dose:

1st cytogenetic assay : 155 and 170 μg/ml , for 3 h exposure time exposure time with a 24 h fixation time with and without metabolic activation system
2nd cytogenetic assay : 100 μg/ml for a 24 h continuous exposure time with a 24 h fixation time and up to 75 μg/ml for a 48 h continuous exposure time with a 48 h fixation time without of S9-mix

Controls

Controls 1

Untreated negative controls:

yes

Remarks:

with vehicule

Negative solvent / vehicle controls:

yes

Remarks:

dimethyl sulfoxide

True negative controls:

no

Positive controls:

yes

Positive control substance:

mitomycin C

Remarks:

Without metabolic activation (-S9-mix)

Controls 2

Untreated negative controls:

yes

Remarks:

with vehicule

Negative solvent / vehicle controls:

yes

Remarks:

dimethyl sulfoxide

True negative controls:

no

Positive controls:

yes

Positive control substance:

cyclophosphamide

Remarks:

With metabolic activation (+S9-mix)

Rationale for test conditions:

Cultured peripheral human lymphocytes were used as test system. Peripheral human lymphocytes are recommended in international guidelines (OECD, EC).
Blood was collected from healthy adult, non-smoking, male volunteers. The Average Generation Time (AGT) of the cells and the age of the donor at the time the AGT was determined (December 2014) are presented below:
Dose range finding study: age 23, AGT = 12.9 h
First cytogenetic assay: age 32, AGT = 12.8 h
Cytogenetic assay 1A: age 25, AGT = 12.8 h
Second cytogenetic assay: age 27, AGT = 12.6 h (24 h exposure time)
age 25, AGT = 12.8 h (48 h exposure time)

Results and discussion

Test results

Key result

Species / strain:

lymphocytes: Human

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

Appropriate toxicity was reached at these dose levels

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

other: all strains/cell types tested

Remarks:

Migrated from field 'Test system'.

Any other information on results incl. tables

At a concentration of 512 µg/ml GR-50-1408 precipitated in the culture medium and was used as the highest concentration of GR-50-1408. In the dose range finding test blood cultures were treated with 5.4, 17, 52, 164 and 512 µg GR-50-1408/ml culture medium with and without S9-mix.

Based on the results of the dose range finding test the following dose levels were selected for the cytogenetic assay:

Without and with S9-mix: 5, 50, 100, 125, 150, 175 and 200 µg/ml culture medium

(3 h exposure time, 24 h fixation time).

Both in the absence and presence of S9-mix, GR-50-1408 did not induce a statistically significant or biologically relevant increase in the number of cells with chromosome aberrations. Both in the absence and presence of S9-mix, GR-50-1408 did not increase the number of polyploid cells and cells with endoreduplicated chromosomes.

To obtain more information about the possible clastogenicity of GR-50-1408, a second cytogenetic assay was performed in which human lymphocytes were continuously exposed to GR-50-1408 in the absence of S9-mix for 24 or 48 hours. The following dose levels were selected for the second cytogenetic assay:

Without S9-mix : 5, 50, 75, 100, 125 and 150 µg/ml culture medium

(24 h and 48 h exposure time, 24 h and 48 h fixation time).

GR-50-1408 did not induce a statistically significant or biologically relevant increase in the number of cells with chromosome aberrations. GR-50-1408 did not increase the number of polyploid cells and cells with endoreduplicated chromosomes

Applicant's summary and conclusion

Conclusions:

Interpretation of results (migrated information): negative - it is concluded that this test is valid and that GR-50-1408 is not clastogenic in human lymphocytes.
Both in the absence and presence of S9-mix GR-50-1408 did not induce a statistically significant or biologically relevant increase in the number of cells with chromosome aberrations in two independent experiments.
No effects of GR-50-1408 on the number of polyploid cells and cells with endoreduplicated chromosomes were observed both in the absence and presence of S9-mix. Therefore it can be concluded that GR-50-1408 does not disturb mitotic processes and cell cycle progression and does not induce numerical chromosome aberrations under the experimental conditions describedt.