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

Long-term toxicity to fish

Administrative data

Endpoint:

fish early-life stage toxicity

Type of information:

experimental study

Adequacy of study:

key study

Study period:

Oct 2013 - NOV 2014

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Study conducted with the base (BAY 73-4605) 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:

2014

Report date:

2014

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 210 (Fish, Early-Life Stage Toxicity 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
- Analytical purity: 100.7%
- Lot/batch No.: BXR4Z07

Sampling and analysis

Analytical monitoring:

yes

Test solutions

Vehicle:

yes

Test organisms

Test organisms (species):

Pimephales promelas

Study design

Test type:

flow-through

Water media type:

freshwater

Limit test:

no

Total exposure duration:

28 d

Remarks on exposure duration:

post hatch (ph)

Results and discussion

Effect concentrations

Effect concentrations 1

Duration:

28 d

Dose descriptor:

NOEC

Effect conc.:

0.007 µg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

test mat.

Basis for effect:

mortality

Effect concentrations 2

Duration:

28 d

Dose descriptor:

LOEC

Effect conc.:

0.025 µg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

test mat.

Basis for effect:

mortality

Any other information on results incl. tables

Table 1: Summary of survival, length and weight of fathead minnow exposed to Regorafenib in an ELS test over 28 days post hatch (ph)

Regorafenib [ug/L] nominal / extrapolated	Hatching rate [%] n=80	Survival day 7 ph [% of hatched larvae]	Survival day 28 ph [% of hatched larvae]	Weight day 28 ph [mg] n=counted larvae	Length day 28 ph [mm] n=counted larvae
0 (control)	98,8	98,7	84,9	24,48 (n=67)	11,22 (n=67)
0 (DMF)	95,0	100	85,3	38,75 (n=65)	12,04 (n=65)
0,03 / 0,01	93,8	98.5	89,3	19,86 (n=67)	10,65 (n=67)
0.1 / 0,03	93,8	95,9	45,0	40,83 (n=34)	12,51 (n=34)
0.3 / 0.08	85,0	96,9	38,2	29,39 (n=26)	11,5 (n=26)
1.0 / 0.38	77,5	93,7	43,5	35,7 (n=27)	12,0 (n=27)
3.0 / 1,39	73,8	94,3	39,4	14,17 (n=23)	8,91 (n=23)

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Conclusions:

The results of this study showed that regorafenib has effects on survival of fish (fathead minnow) at concentrations from 0.025 ¿g/L onwards.
Thus, the overall no observed effect concentration (NOEC) was 0.007 ¿g/L, the lowest observable effect concentration (LOEC) was 0.025 ¿g/L.

Executive summary:

The purpose of this study was to determine the aquatic toxicity of Regorafenib in the early-life-stage test on the fathead minnow (Pimephales promelas). Regorafenib is a multi-kinase inhibitor developed for tumor therapy. The study was conducted following the OECD guideline 210.

Freshly fertilized eggs (80 per treatment group, 4 replicates) from a laboratory culture of adult fathead minnow were used for each test solution of regorafenib and for the control group. For each group two separate glass vessels, each with 20 eggs, were introduced.

The eggs and larvae were exposed for a period of 28 days post-hatch (ph) to five test solutions of regorafenib with nominal concentrations of 0.03 ¿g/L, 0.1 ¿g/L, 0.3 ¿g/L, 1 ¿g/L und 3 ¿g/l. Additionally a tap water control and a solvent control (DMF) was used under the same conditions. For the preparation of the stock solutions, the test substance was dissolved in DMF.

Mortalities and visible abnormalities were recorded daily over the exposure phase. Hatching success and survival were recorded during the first phase of embryonic development as well as growth and survival during the larval stage until the end of the exposure period. The hydrographic parameters were recorded daily and after day 7 ph weekly.

Fish larvae hatched 6 days after the introduction of the eggs in all test solutions. The hatching success was >95% in both the water and solvent controls, and decreased concentration related in all test solutions from 94 to 74%. This was, however, not statistically significant. For weight increase, there was a decrease compared to control fish, however, this was also not statistically significant. The NOEC and LOEC for length increase was 0.34 ¿g/L and 1.39 ¿g/L.

The number of dead fish increased with increasing concentration. For post-hatch survival the NOEC was 0.007 ¿g/L and the LOEC was 0.025 ¿g/L.