D-threo-Pentitol, 2,5-anhydro-1,3,4-trideoxy-2-C-(2,4-difluorophenyl)-4-[[4-[4-[4-[[[1-[(1S,2S)-1-ethyl-2-(phenylmethoxy)propyl]hydrazino]carbonyl]amino]phenyl]-1-piperazinyl]phenoxy]methyl]-1-(1H-1,2,4-triazol-1-yl)-

Genetic toxicity: in vitro

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:

10 November to 01 December, 1998

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Study conducted according to OECD method and in accordance with GLP. Study material is well characterized.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1998

Report date:

1999

Materials and methods

Test guideline

Test guideline 1

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Test guideline 2

Qualifier:

according to guideline

Guideline:

other: Japan's MITI, MHW, MOL and MAFF.

GLP compliance:

yes

Remarks:

OECD ENV/MC/CHEM(98)17

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

2,5-anhydro-4-{[4-(4-{4-[({1-[(2S,3S)- 2-(benzyloxy)pentan-3-yl]hydrazino}carbonyl)amino]phenyl}piperazin1-yl)phenoxy]methyl}-1,3,4-trideoxy-2-(2,4-difluorophenyl)-1-(1H-1,2,4-triazol-1-yl)-

EC Number:

701-059-1

Cas Number:

345217-03-0

Molecular formula:

C43H50F2N8O4

IUPAC Name:

2,5-anhydro-4-{[4-(4-{4-[({1-[(2S,3S)- 2-(benzyloxy)pentan-3-yl]hydrazino}carbonyl)amino]phenyl}piperazin1-yl)phenoxy]methyl}-1,3,4-trideoxy-2-(2,4-difluorophenyl)-1-(1H-1,2,4-triazol-1-yl)-

Details on test material:

WHITE SOLID

Method

Species / strain

Species / strain 1

Species / strain / cell type:

S. typhimurium TA 1535, TA 1537, TA 98 and TA 100

Species / strain 2

Species / strain / cell type:

E. coli WP2 uvr A

Metabolic activation:

with and without

Metabolic activation system:

S-9 mix from Aroclor 1254 induced rat liver

Test concentrations with justification for top dose:

For genotoxicity experiment concentrations (with & without metabolic activation) used:
Preliminary Toxicity Study: 0, 0.15, 0.5, 1.5, 5, 15, 50, 150, 500, 1500 and 5000 µg/plate.
Mutation Study: 0, 50, 150, 500, 1500 and 5000 µg/plate

Vehicle / solvent:

solvent- dimethyl sulphoxide.

Controls

Controls 1

Positive control substance:

4-nitroquinoline-N-oxide

9-aminoacridine

N-ethyl-N-nitro-N-nitrosoguanidine

benzo(a)pyrene

other: 2-Aminoanthracene

Controls 2

Negative solvent / vehicle controls:

yes

Remarks:

DMSO

Details on test system and experimental conditions:

S9 was prepared from the livers of male Sprague-Dawley rats weighing ~ 250g. Each received a single i.p. injection of Aroclor 1254 at 500 mg/kg. The S9 was checked for suitability using the indirect mutagens 2AA and BP.

Preliminary Toxicity Study, Mutation Study 1 and Mutation Study 2 test plates were incubated at 37°C for 48 hrs and assessed for numbers of relevant colonies using a Domino colony counter. Manual counts were performed at and above 1500 µg/plate due to test material precipitation.

Toxicity: No toxicity was exhibited to any of the strains of bacteria used.
A white powdery precipitate was observed at and above 1500µg/plate. This did not prevent the scoring of revertant colonies.

Evaluation criteria:

Evaluation criteria: test article would be considered mutagenic if:
The test material should have induceda reproducible, dose-related and statistically (Dunnett's method of linear regression) significant increase in the revertant count in at least one strain of bacteria.
If a greater than twofold increase in revertant count is observed in two experiments then this is taken as evivence of a positive response.

Statistics:

Mean and standard deviation of the plate counts for each treatment were determined

Results and discussion

Test results

Test results 1

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

Untreated negative controls validity:

valid

Positive controls validity:

valid

Test results 2

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

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

Observations:
All positive control chemicals used in the test induced marked increases in the frequency of revertant colonies, both with and without metabolic activation. Thus, the sensitivity of the assay and the efficacy of the S9 mix were validated.

Applicant's summary and conclusion

Conclusions:

Interpretation of results (migrated information):
negative

No dose-related and reproducible increases in revertant colony frequency were observed in any tester strains at any concentration, both with and without S9. The test material was considered to be non-mutagenic under the conditions of this test.

Executive summary:

No toxicity was observed to any of the strains of bacteria used.

No significant increases in the frequency of revertant colonies were recorded for any of the strains of bacteria, at any dose level either with or without metabolic activation.

All of the positive control chemicals used in the test induced marked increases in the frequency of revertant colonies thus confirming the activity of the S9 -mix and the sensitivity of the bacterial strains.