Registration Dossier

Administrative data

Endpoint:
short-term repeated dose toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Study period:
12/01/2016 - 14/04/2016
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2016
Report Date:
2016

Materials and methods

Test guideline
Qualifier:
equivalent or similar to
Guideline:
OECD Guideline 407 (Repeated Dose 28-Day Oral Toxicity in Rodents)
Deviations:
no
GLP compliance:
yes (incl. certificate)

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
liquid

Test animals

Species:
rat
Strain:
Crj: CD(SD)
Sex:
male/female

Administration / exposure

Route of administration:
oral: gavage
Details on oral exposure:
Dose levels:

0 (control), 40, 200, and 1000 mg/kg/day

Reason for dose selection:

In the dose range-finding study in which the test article dissolved in corn oil was administered by oral gavage at 40, 200, and 1000 mg/kg/day to 3 male and 3 female SD rats repeatedly for 7 days, no test article-related changes were noted at any dose level. Therefore, 1000 mg/kg was selected as the high dose, and 200 and 40 mg/kg were selected as the middle and low doses, respectively, with a common ratio of 5. Recovery groups were set for the control and high dose groups to investigate the reversibility during 14-day withdrawal following 28-day administration.
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
Analytical method - Gas chromatograph
Column - DB-1301
Injection temp - 240°C
Detection temp. - 250°C
Column condition - Initial 40°C (10min hold), Rate lO°C/min, Final 240°C 15min
Frequency of treatment:
Once daily for 28 days
Doses / concentrationsopen allclose all
Dose / conc.:
40 mg/kg bw/day (nominal)
Dose / conc.:
200 mg/kg bw/day (nominal)
Dose / conc.:
1 000 mg/kg bw/day (nominal)
No. of animals per sex per dose:
6
Control animals:
yes, concurrent no treatment

Examinations

Observations and examinations performed and frequency:
The first day of administration was designated as Administration Day 1 (Day 1 ), and the day after Administration Day 28 as Recovery Day 1, and the day of necropsy was the day after Administration Day 28 (Day 29) or the day after Recovery Day 14 (Recovery Day 15). Administration Days 1 to 7 were designated as Administration Week 1, and subsequent days were counted on a 7 day-basis as follows: Administration Days 22 to 28 was Administration Week 4 (final week of administration), Recovery Days 1 to 7 was Recovery Week 1, and Recovery Days 8 to 14 was Recovery Week 2 (final week of recovery).
Sacrifice and pathology:
Animals were observed for external appearance, and blood was collected from the abdominal aorta under anesthesia with pentobarbital sodium. Animals were then euthanized by exsanguination and all organs and tissues were macroscopically observed. The organs and tissues described below were fixed and preserved in 10% neutral buffered formalin. The eyeballs and harderian glands were fixed and preserved in Davidson's fixative. The testes and epididymides were fixed in Bouin's solution and preserved in 70% ethanol. The lung was immersed in the fixative after infusing fixative into it. For bilateral organs, both sides of them were fixed and preserved.
Statistics:
Statistical analyses were performed using the computer system (MiTOX). The data during administration period obtained from the toxicity study groups and those from the recovery study groups were summed for statistical analyses.

Results and discussion

Results of examinations

Clinical signs:
effects observed, treatment-related
Description (incidence and severity):
Clinical signs:

Malocclusion that was likely attributable to an accident in the cage in 1 female in the control group. During recovery period, no abnormalities were noted in clinical observation in males or females in any treated group.

Detailed clinical observation

During administration and recovery periods, no significant differences were noted in any parameters in males or females in any treated group compared with the control group.

Functional observations

During Administration Week 4, motor activity (30-40, 40-50, and 50-60 min) and grip strength of the forelimbs of males in the 1000 mg/kg group were significantly
lower than those in the control group. No significant differences were noted in any parameters in males or females in the 40 or 200 mg/kg group. During Recovery Week 2, motor activity (0-10, 10-20, 30-40, and 0-60 min) of females in the 1000 mg/kg group was significantly high. This change was not noted during Administration Week 4, and was considered unrelated to the test article administration. No significant differences were noted in motor activity in males or sensory and motor reactivity to stimuli or grip strength of males or females compared with the control group.
Mortality:
mortality observed, non-treatment-related
Description (incidence):
During administration period, 1 male (animal No. 10301) in the 200 mg/kg group was found dead before dosing on Administration Day 20.
Body weight and weight changes:
effects observed, non-treatment-related
Description (incidence and severity):
No significant differences were noted in body weight or body weight gain during administration period in males or females in any treated group compared with the control group. During recovery period, body weight gain in females in the 1000 mg/kg group was significantly low. This was considered toxicologically insignificant because no significant differences were noted in body weight. In males, no significant differences were noted compared with the control group.
Food consumption and compound intake (if feeding study):
no effects observed
Description (incidence and severity):
There were no significant differences in food consumption during administration or recovery period in males or females in any treated group compared with the control group.
Food efficiency:
not examined
Water consumption and compound intake (if drinking water study):
not examined
Ophthalmological findings:
not examined
Haematological findings:
effects observed, non-treatment-related
Description (incidence and severity):
In the toxicity study groups (at the end of administration), no significant differences were noted in any parameter in males or females in any treated group compared with the control group. In the recovery study groups (at the end of recovery), WBC and lymphocyte were significantly low in males and PT was significantly shortened in females in the 1000 mg/kg group. These changes were not noted at the end of administration and are considered toxicologically insignificant.
Clinical biochemistry findings:
effects observed, non-treatment-related
Description (incidence and severity):
In the toxicity study groups ( at the end of administration), T-Bil in males in the 40 and 1000 mg/kg groups was significantly lower than that in the control group; however, individual values were within the range of historical control data (mean 0.049 mg/dL, range 0.029 to 0.069 mg/dL). Since the low values have little clinical significance, this change was considered toxicologically insignificant. TG in males 2) in the 200 mg/kg group was significantly low, which was considered unrelated to the test article administration because no dose relationship was noted. No significant difference was noted in any parameter in females compared with the control group. In the recovery study groups, TG was significantly low and IP was significantly high in females in the 1000 mg/kg group. These changes were not noted at the end of administration and individual values were within the ranges of historical control data" (TG, mean 13.2 mg/dL, range 2.2 to 24.2 mg/dL; IP, mean 7.51 mg/dL, range 5.35 to 9.67 mg/dL) or slightly deviated from the ranges; therefore, these changes were considered toxicologically insignificant. In males, no significant differences were noted in any parameter compared with the control group.
Urinalysis findings:
no effects observed
Description (incidence and severity):
There were no significant differences in any parameters during Administration Week 4 or Recovery Week 2 in males or females in any treated group compared with the control group.
Behaviour (functional findings):
not examined
Immunological findings:
not examined
Organ weight findings including organ / body weight ratios:
effects observed, non-treatment-related
Description (incidence and severity):
In the recovery study groups, absolute testis weight and relative prostate weight were significantly low in males and relative adrenal weight was significantly high in females in the 1000 mg/kg group. These changes were considered toxicologically insignificant because no changes related to the test article administration were noted in the weight or histopathology of these organs at the end of administration, and individual values were within the ranges of historical control data
(testis, mean 3.108 g, range 2.542 to 3.674 g; prostate, mean 145.993 mg/100 g, range 81.573 to 210.413 mg/100 g; adrenal, females, mean 31.336 mg/100 g, range 21.130 to 41.542 mg/100 g).
Gross pathological findings:
effects observed, non-treatment-related
Description (incidence and severity):
Necropsy at the end of administration in the toxicity study groups revealed dilatation of the renal pelvis in 1 male and malocclusion in 1 female in the control group.
Other than these, no abnormalities were noted in any animal. The male that died in the 200 mg/kg group (animal No. 10301) showed no abnormalities. Necropsy in the recovery study groups revealed no abnormalities in males or females in any treated groups.
Neuropathological findings:
not examined
Histopathological findings: non-neoplastic:
effects observed, non-treatment-related
Description (incidence and severity):
In the toxicity study groups, histopathological findings including microgranuloma in the liver and basophilic change of the renal tubule in the kidneys were sporadically noted in males or females in the 1000 mg/kg group. These changes were noted also in the control group or historical control data", and of low frequency; therefore, these were determined to be incidental findings. The male that died in the 200 mg/kg group (animal No. 10301) showed slight alveolar and perivascular edema in the lung.
Histopathological findings: neoplastic:
not examined
Other effects:
not examined

Effect levels

Key result
Dose descriptor:
NOAEL
Effect level:
200 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
behaviour (functional findings)

Target system / organ toxicity

Critical effects observed:
no

Applicant's summary and conclusion

Conclusions:
A repeated dose oral toxicity study was conducted in which HCFO-1233yd(Z) was administered to 6 males and 6 females per group of Crl:CD (SD) rats by oral gavage at O (control: com oil), 40, 200, and 1000 mg/kg once daily for 28 days. Recovery study groups at doses of O and 1000 mg/kg were separately employed, and during the 14-day withdrawal period following the end of 28-day administration, reversibility of the toxicity that occurred was investigated in 6 males and 6 females per group.

In the 40 mg/kg group, no changes related to the test article administration were noted in clinical observation, body weight, food consumption, detailed clinical observation, functional observations, urinalysis, hematology, biochemistry, necropsy, or organ weight in males or females.

In the 200 mg/kg group, 1 male died on Administration Day 20. Histopathological examination of this animal revealed slight alveolar and perivascular edemas in the lung, which indicated circulatory disorder. This was considered unrelated to the test article administration because this change was not noted in the 1000 mg/kg group. No effects of the test article administration were noted in any of the other parameters in males or females.

In the 1000 mg/kg group, motor activity and grip strength of the forelimbs were significantly low during Administration Week 4 in males. In the acute toxicity study of HCFO-1233yd(Z), staggering gait was noted at 2000 mg/kg; therefore, the changes noted in the present study were considered related to the test article administration. This change was not noted during the 2 recovery weeks, indicating reversibility of the change. There were no changes related to the test article administration in males or females in any other examination parameters including histopathology.

As described above, administration of 1000 mg/kg HCFO-1233yd(Z) resulted in decreases in motor activity and grip strength of the forelimbs in males. While in females, no toxic changes related to the test article administration were noted in clinical observation, detailed clinical observation, functional observations, body weight, food consumption, urinalysis, hematology, biochemistry, necropsy, organ weight, or histopathology at doses up to 1000 mg/kg.
Executive summary:

On the basis of these results, the No-Observed Adverse Effect Level (NOAEL) of HCFO-1233yd(Z) was determined to be 200 mg/kg/day for males and 1000 mg/kg/day for females under the present study conditions.