2-Pyridinecarboxamide, 4-[4-[[[[4-chloro-3-(trifluoromethyl)phenyl]amino]carbonyl]amino]-3-fluorophenoxy]-N-methyl-, hydrochloride (1:1)

Long-term toxicity to aquatic invertebrates

Administrative data

Endpoint:

long-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

key study

Study period:

Nov 2012 - Jan 2013

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Study conducted with the base (BAY 73-4506) according to OECD guideline under GLP

Cross-reference

Cross-reference 1

Reason / purpose for cross-reference:

reference to same study

Cross-reference 2

Reason / purpose for cross-reference:

reference to other study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2013

Report date:

2013

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 211 (Daphnia magna Reproduction Test)

Deviations:

no

Principles of method if other than guideline:

not relevant

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

4-(4-{3-[4-Chloro-3-(trifluoromethyl)phenyl]ureido}-3-fluorophenoxy)- N-methylpyridine-2-carboxamide

IUPAC Name:

4-(4-{3-[4-Chloro-3-(trifluoromethyl)phenyl]ureido}-3-fluorophenoxy)- N-methylpyridine-2-carboxamide

Details on test material:

- Name of test material (as cited in study report): Regorafenib (BAY 73-4506)
- Analytical purity: 100%
- Lot/batch No.: BXR4Z07

Sampling and analysis

Analytical monitoring:

yes

Test solutions

Vehicle:

yes

Test organisms

Test organisms (species):

Daphnia magna

Study design

Test type:

semi-static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

21 d

Results and discussion

Effect concentrations

Effect concentrations 1

Duration:

21 d

Dose descriptor:

NOEC

Effect conc.:

24.7 µg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

test mat.

Basis for effect:

reproduction

Effect concentrations 2

Duration:

21 d

Dose descriptor:

NOEC

Effect conc.:

10.5 µg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

test mat.

Basis for effect:

immobilisation

Effect concentrations 3

Duration:

21 d

Dose descriptor:

LOEC

Effect conc.:

57.6 µg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

test mat.

Basis for effect:

reproduction

Effect concentrations 4

Duration:

21 d

Dose descriptor:

LOEC

Effect conc.:

24.7 µg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

test mat.

Basis for effect:

immobilisation

Any other information on results incl. tables

Table 1: Cumulative number of off-spring and number of brood per surviving animals after 21 days of exposure to Regorafenib (mean values ± standard deviation)

Nominal	Number of surviving	No. of off-spring per	Number of broods per
concentration of	animals	surviving parent	surviving parent
BAY 73-4506	(survival / total)	animals	animal
[microg/L]		[MV ± SD]	[MV ± SD]
0 (control)	10/10	74.0 ± 7.4	4.8 ± 0.6
0 (vehicle control)	8/10	68.3 ± 13.0	4.4 ± 0.7
2.6	9/10	71.0 ± 10.6	4.7 ± 0.5
5.5	9/10	67.8 ± 14.7	4.8 ± 0.4
10.5	9/10	68.4 ± 5.1	5.1 ± 0.3
24.7	4/10	64.8 ± 18.0	4.8 ± 0.5
57.6	1/10	33.0	3.0

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Conclusions:

Regorafenib had a concentration related mortality and effects on reproduction in Daphnia magna. The most sensitive NOEC for chronic
immobilization was 10.5 microg/L, the LOEC was 24.7 microg/L.

Executive summary:

The purpose of this study was to determine the chronic toxicity of the compound Regorafenib (BAY 73-4506) on reproduction of Daphnia magna in the context of evaluating the risk to the aquatic environment. Regorafenib is developed for cancer therapy.

The study was conducted in agreement with the guideline of the OECD, no. 211. Due to the low water solubility of Regorafenib, the study was conducted with DMF as vehicle.

10 Daphnia magna were used for each of the 5 test concentrations of Regorafenib and 10 for the dilution water control and the vehicle control. The Daphnia were exposed to the test solution, the vehicle control and the tap water for a period of 21 days (start of treatment = day 1) under semistatic conditions, i.e. the solutions were renewed three times a week. The Daphnia were held individually. The number of living off-spring was recorded at each change of test solution 3 times per week, immobilization of parent animals was recorded daily.

For the preparation of the stock solution an amount of 100 mg Regorafenib was dissolved in 100 mL DMF by stirring. Aliquots of the obtained stock solution were further diluted with tap water and DMF in order to prepare the appropriate test concentrations: 6.3 microg/L, 12.5 microg/L, 25 microg/L, 50 microg/L and 100 microg/L. The vehicle control and the test solutions contained 0.1 mL DMF/L in water.

The test temperature was in the range of 19.9°C to 21.6°C. The pH was in a range between 7.6 and 8.6, and the oxygen concentration between 7.7 and 9.7 mg/L. The light/dark rhythm was adjusted to 16 hours/8 hours.

Samples for the concentration analysis by HPLC/MS-detection were taken weekly from the test and control solutions and samples from the stock solution were taken at the beginning and at the end of the study.

The mean number of off-spring per parent animal varied between 74.0 and 33.0, the number of broods varied between 4.8 and 3.0 showing a dose dependence. The first brood in the control and the vehicle control group was recorded at day 9 (mean) of the exposure.

Regorafenib had a concentration related mortality and effects on reproduction in Daphnia magna.