3-Quinolinecarboxylic acid, 1-cyclopropyl-6,8-difluoro-1,4-dihydro-7-[(4aS,7aS)-octahydro-6H-pyrrolo[3,4-b]pyridin-6-yl]-4-oxo-

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:

other information

Study period:

August-October 1997

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Well reported GLP study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1997

Report date:

1997

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method B.13/14 (Mutagenicity - Reverse Mutation Test Using Bacteria)

Test guideline 3

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.5265 (The Salmonella typhimurium Bacterial Reverse Mutation Test)

GLP compliance:

yes (incl. QA statement)

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

7-[(4aS,7aS)-1,2,4a,5,7,7a-hexahydropyrrolo[3,4-b]pyridin-6-yl]-1-cyclopropyl-6,8-difluoro-4-oxo-3H-quinoline-3-carboxylic acid

EC Number:

604-776-7

Cas Number:

151213-15-9

Molecular formula:

C20 H21 F2 N3 O3

IUPAC Name:

7-[(4aS,7aS)-1,2,4a,5,7,7a-hexahydropyrrolo[3,4-b]pyridin-6-yl]-1-cyclopropyl-6,8-difluoro-4-oxo-3H-quinoline-3-carboxylic acid

Details on test material:

BAY z 7906, yellowish powder, batch 501779, content 99.8%

Method

Target gene:

histidin gene locus

Species / strain

Species / strain / cell type:

S. typhimurium, other: TA 1535, TA 100, TA 1537, TA 98 and TA 102

Metabolic activation:

with and without

Metabolic activation system:

S9-mix

Test concentrations with justification for top dose:

BAY z 7906: 0.00005-5000 µg/plate or tube (first test 50-5000 µg/plate, due to toxicity of the substance doses from 0.5-16 µg/plate and 0.00005-0.016 µg/plate were chosen for the repeat tests)
Sodium-azide: 10 µg/plate (TA 1535)
Nitrofurantoin: 0.2 µg/plate (TA 100)
4-Nitro-1,2-phenylene diamine: 10 µg/plate (TA 1537), 0.5 µg/plate (TA 98)
Cumene: 50 µg/plate (TA 102)
2-Aminoanthracene: 3 µg/plate

Controls

Untreated negative controls:

yes

Negative solvent / vehicle controls:

no

Remarks:

No solvent control was used since sufficient evidence was available in the literature and from testing laboratory experience, indicating that the solvents used had no influence on the spontaneous mutant counts of the used strains.

Positive controls:

yes

Positive control substance:

other: Sodium-azide, Nitrofurantoin, 4-Nitro-1,2-phenylene diamine, Cumene, 2-Aminoanthracene

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:

cytotoxicity

Remarks:

up to and including 0.0016 µg/plate no bacteriotoxic effects; at higher doses strong, strain-specific bacteriotoxic effect, so that this range could only be used to a limited extent up to and including 0.016 µg/plate for assessment purposes

Untreated negative controls validity:

valid

Positive controls validity:

valid

Test results 2

Species / strain:

S. typhimurium TA 102

Metabolic activation:

with and without

Genotoxicity:

positive

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

up to and including 0.0016 µg/plate no bacteriotoxic effects; at higher doses strong, strain-specific bacteriotoxic effect, so that this range could only be used to a limited extent up to and including 0.016 µg/plate for assessment purposes

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

Strain TA 102 revealed in the plate incorporation test a dose–related increase in mutant counts to well over double those of negative controls. The findings for Salmonella typhimurium TA 102 were confirmed by the results of the preincubation trial. The lowest dose at which this finding was reproducible was 0.005 μg per plate for Salmonella typhimurium TA 102. Positive findings were obtained with and without S9 mix, the effects being comparable in both cases.

Summary of the Results with Bay Z 7906 in the Salmonella/Microsome Test

S9 mix	TA 1535	TA 100	TA 1537	TA 98	TA 102
without	–ve	–ve	–ve	–ve	+ve
with	–ve	–ve	–ve	–ve	+ve

–ve = negative, +ve = positive

Applicant's summary and conclusion

Executive summary:

Bay Z 7906 was investigated using the Salmonella/microsome plate incorporation and preincubation test for point-mutagenic effects in doses up to and including 5000 µg/plate or tube on the five histidine-auxotrophic Salmonella typhimurium LT2 strains TA 1535, TA 100, TA 1537, TA 98 and TA 102. Doses up to and including 0.0016 μg/plate did not cause any bacteriotoxic effects. No

inhibition of growth was noted. Higher doses had a strong, strain–specific bacteriotoxic effect. Therefore they could only partly be used for assessment purposes up to and including 0.016 μg per plate.

Evaluation of individual dose groups, with respect to relevant assessment parameters (dose effect, reproducibility), revealed biologically relevant variations from the respective negative controls for TA 102 without and with S9 mix. The lowest reproducible effective dose was 0.005 µg/plate. The Salmonella/microsome test thus showed Bay Z 7906 to induce mutations in the S. typhimurium strain TA 102.