Registration Dossier

Administrative data

Endpoint:
screening for reproductive / developmental toxicity
Type of information:
experimental study
Adequacy of study:
key study
Study period:
From July 26, 2016 to January 23, 2017
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
other company data
Title:
Unnamed
Year:
2017
Report Date:
2017

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 421 (Reproduction / Developmental Toxicity Screening Test)
GLP compliance:
yes

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
solid: particulate/powder
Remarks:
migrated information: powder
Details on test material:
- Name of test material (as cited in study report): Everdirect SH12
- Substance type: Powder
- Composition of test material, percentage of components: 69.96 %
- Lot/batch No.: 3506
- Storage condition of test material: Ambient

Test animals

Species:
rat
Strain:
other: Crl: CD(SD)
Sex:
male/female
Details on test animals and environmental conditions:
- Source: BioLASCO Taiwan Co., Ltd., Taipei, Taiwan
- Age at study initiation: about 9-week old
- Weight at study initiation: Males: 322-375 g; Females: 206-249 g
- Housing: Except for the mating period, animals were housed individually in polycarbonate cages. While mating, animals were pair-housed in stainless steel wire mesh cages.
- Diet: ad libitum
- Water: ad libitum
- Acclimation period: 11 days
- Temperature (°C): 21 ± 2 °C
- Humidity (%): 50 ± 20%
- Photoperiod: 12-hrs dark / 12-hrs light

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
corn oil
Doses / concentrationsopen allclose all
Dose / conc.:
0 mg/kg bw/day (nominal)
Remarks:
Dose group 1
Dose / conc.:
100 mg/kg bw/day (nominal)
Remarks:
Dose group 2
Dose / conc.:
300 mg/kg bw/day (nominal)
Remarks:
Dose group 3
Dose / conc.:
1 000 mg/kg bw/day (nominal)
Remarks:
Dose group 4
No. of animals per sex per dose:
For male: ten
For female: ten
Control animals:
yes, concurrent vehicle

Results and discussion

Results: P0 (first parental animals)

General toxicity (P0)

Clinical signs:
effects observed, non-treatment-related
Description (incidence and severity):
All animals survived the study period. Test article-related clinical observations were purple or red-brown colored feces, brown, pink or purple colored hair stained on forelimbs/jaw/mouth and salivation. The noisy respiration at the high dose level was observed at low incidence (only one male rat) and low frequency (only observed on six days). One low dose female showed mass on right side.
Overall, no death was observed in males and females during the study period. Dosedependent
clinical observations were test article-like colored feces, test article-like
colored hair stained and salivation.
Mortality:
no mortality observed
Body weight and weight changes:
effects observed, non-treatment-related
Description (incidence and severity):
In females, statistically significantly increased body weight gain compared to the controls was observed in females of Group 4 on D8. There was no statistically significant differences in body weight and body weight gain changes in treated groups compared to control animals in males.
Overall, the test article did not cause change in body weight gain in males and females in premating, gestation and lactation phases.
Food consumption and compound intake (if feeding study):
effects observed, non-treatment-related
Description (incidence and severity):
In females, statistically significantly increased food consumption compared to the controls was observed in females of Group 4 on D8. There was no statistically significant difference between treated and control groups in males.
Overall, the test article did not cause change in food intake in premating of males and females or; in the gestation and lactation phases of females.

Reproductive function / performance (P0)

Reproductive function: oestrous cycle:
no effects observed
Description (incidence and severity):
There were no statistically significant differences in the number of corpora lutea and implantations between control and treated groups. Pre-implantation and pre-natal losses were observed in all groups including the controls. Decreased size of thymus was observed in one animal of Group 2 and two animals of Group 4.Overall, there were no test article-related effects in female gross and uterus examinations, or microscopic findings in the study animals examined.
Reproductive function: sperm measures:
no effects observed
Description (incidence and severity):
There were no statistically significant differences in testis and epididymis weights between control and treated groups. No gross changes were observed in any males.
Reproductive performance:
effects observed, non-treatment-related
Description (incidence and severity):
In males, a positive indication of mating was obtained for 9, 8, 9 and 9 of 10 animals in Groups 1 to 4, resulting in a mating index of 90%, 80%, 90% and 90%, respectively.
In females, a positive indication of mating was obtained for 10 of 10 animals in Groups 1 and 4 and for 9 of 10 animals in Groups 2 and 3. The mating index was
100%, 90%, 90% and 100% for Group 1 to 4, respectively.
Overall, the test article did not cause changes in mating indices of male and female, fertility indices, pre-coital intervals and pregnancy days.

Effect levels (P0)

Dose descriptor:
NOAEL
Effect level:
ca. 1 000 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
mortality
reproductive function (oestrous cycle)
reproductive function (sperm measures)
reproductive performance

Results: F1 generation

General toxicity (F1)

Clinical signs:
not examined
Mortality / viability:
mortality observed, non-treatment-related
Description (incidence and severity):
All pregnant rats delivered live pups. The gestation index was 100% in all groups. One dead pup was observed at birth in Groups 2 and 3. The incidence of dams with dead pups at birth was 0%, 13%, 13% and 0% in Group 1, 2, 3 and 4, respectively. On P4, a decreased number of live pups was observed in all groups. The 4-day survival index was 99.3%, 96.7%, 96.6% and 98.1% in Groups 1 to 4, respectively. No external abnormally pups were observed at birth in the control and treated groups. At birth, 44.6% to 51.9% of pups were male and on P4, 47.0% to 56.5% pups were male. The gender difference at birth and on P4 was a consequence of difficult sex determination at birth. The litter weights in all groups increased from P0 to P4.

Effect levels (F1)

Dose descriptor:
NOAEL
Generation:
F1
Based on:
test mat.
Remarks on result:
not measured/tested

Overall reproductive toxicity

Reproductive effects observed:
no

Any other information on results incl. tables

Dose Formulation Analysis

For the difference between the mean value and the targeted concentration, exception of the dose group 3 prepared on the first preparation day, other formulations were within ± 15.0% on the first and last preparation. The dose group 3 on first preparation day was calculated as 354 mg/kg and higher than the study design (300 mg/kg/day). The homogeneity verification revealed that the precision of samples on the first preparation for the dose group 2 was 17.4% and on the first and last preparation for other doses formulation were ≤ 10.0%. The dose level in Group 2 prepared on first preparation day may be up to 111 mg/kg and higher than the study design (100 mg/kg/day).

 

Mortality and Clinical Observations

All animals survived the study period. Test article-related clinical observations were purple or red-brown colored feces, brown, pink or purple colored hair stained on forelimbs/jaw/mouth and salivation. The noisy respiration at the high dose level was observed at low incidence (only one male rat) and low frequency (only observed on six days). One low dose female showed mass on right side.

 

Body Weights

In females, statistically significantly increased body weight gain compared to the controls was observed in females of Group 4 on D8. There was no statistically significant differences in body weight and body weight gain changes in treated groups compared to control animals in males.

 

Food Consumption

In females, statistically significantly increased food consumption compared to the controls was observed in females of Group 4 on D8. There was no statistically significant difference between treated and control groups in males.

 

Cohabitation and Pregnancy

In males, a positive indication of mating was obtained for 9, 8, 9 and 9 of 10 animals in Groups 1 to 4, resulting in a mating index of 90%, 80%, 90% and 90%, respectively. In females, a positive indication of mating was obtained for 10 of 10 animals in Groups 1 and 4 and for 9 of 10 animals in Groups 2 and 3. The mating index was 100%, 90%, 90% and 100% for Group 1 to 4, respectively. There was one animal in Group 1, 2 and 4, respectively, that showed a pre-coital interval longer 5 days. In each of these females, the mating with the first male failed but the copulation with another male was successful. In Groups 1 to 3, 1 animal with positive indications of mating was later found not to be pregnant. Thus, the fertility index was 90%, 89%, 89% and 100% in the control, low, mid and high dose group, respectively. The average length of gestation was 21.9 − 22.1 days without dose-dependency.

 

Litter Observation

All pregnant rats delivered live pups. The gestation index was 100% in all groups. One dead pup was observed at birth in Groups 2 and 3. The incidence of dams with dead pups at birth was 0%, 13%, 13% and 0% in Group 1, 2, 3 and 4, respectively. On P4, a decreased number of live pups was observed in all groups. The 4-day survival index was 99.3%, 96.7%, 96.6% and 98.1% in Groups 1 to 4, respectively. No external abnormally pups were observed at birth in the control and treated groups. At birth, 44.6% to 51.9% of pups were male and on P4, 47.0% to 56.5% pups were male. The gender difference at birth and on P4 was a consequence of difficult sex determination at birth. The litter weights in all groups increased from P0 to P4.

 

Male Gross Examination, Organ Weight and Histopathology

There were no statistically significant differences in testis and epididymis weights between control and treated groups. No gross changes were observed in any males.

 

Females Gross, Uterus Examination and Histopathology

There were no statistically significant differences in the number of corpora lutea and implantations between control and treated groups. Pre-implantation and pre-natal losses were observed in all groups including the controls. Decreased size of thymus was observed in one animal of Group 2 and two animals of Group 4.

Applicant's summary and conclusion

Conclusions:
According to OECD 421 test method, the no observed adverse effect level of Everdirect SH12 in rat’s reproductive performance was 1000 mg/kg/day.
Executive summary:

This test using the procedures outlined in the QPS Study Plan for T65315034-RP and OECD 421. Everdirect SH12 (100, 300 or 1000 mg/kg/day) was given orally by gavage once daily to male rats for 28 days and female rats for 40 to 53 days, depending on the time of copulation and gestation. Dose-related clinical observations in the treated groups includedtest article-like colored feces, test article-like colored hair stained and salivation. The test article did not cause change in body weight, food consumption in premating, gestation and lactation phases. There were no test article-related effects in mating index of both gender, and fertility index, pre-coital interval and pregnancy day in the females. There were no treatment related effects on pups viability, body weight, sex and gross external examinations. Macroscopic and microscopic evaluations did not reveal treatment-related effects in male and female fertility or reproductive organs. The no observed adverse effect level of Everdirect SH12 in rat’s reproductive performance was 1000 mg/kg/day.