Registration Dossier

Administrative data

Endpoint:
two-generation reproductive toxicity
Remarks:
based on test type (migrated information)
Type of information:
experimental study
Adequacy of study:
key study
Study period:
11 February 2013 - 12 July 2013
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Study performed according to OECD Guideline and in compliance with GLP.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2013
Report Date:
2013

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 421 (Reproduction / Developmental Toxicity Screening Test)
Deviations:
no
GLP compliance:
yes (incl. certificate)

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
other: liquid
Details on test material:
- Name of test material (as cited in study report): MTF (7.8-benzotricyclo[4.3.0.1(2.5)]deca-3-ene)
- Substance type: Organic
- Physical state: Colourless liquid
- Analytical purity:100%
- Lot/batch No.: M1204
- Expiration date of the lot/batch: End of December 2015
- Storage condition of test material: Dark, room temperature, under nitrogen

Test animals

Species:
rat
Strain:
other: Crl: CD(SD)
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: reputable commercial supplier
- Age at study initiation: ca 71 days old
- Weight at study initiation: Males: 336-390 g; Females: 234-284 g;
- Fasting period before study: no
- Housing: Cages comprised of a polycarbonate or polypropylene body with a stainless steel mesh lid; changed at appropriate intervals.
Solid (polycarbonate) bottom cages were used during the acclimatisation, pre-pairing, gestation, littering and lactation periods.
Polypropylene cages with a grid bottom were used during pairing. These were suspended above absorbent paper which was changed daily.
- Diet: ad libitum
- Water: ad libitum
- Acclimation period: 6 days before commencement of treatment

ENVIRONMENTAL CONDITIONS
- Temperature (°C) and Humidity (%): Monitored and maintained within the range of 19-23 ºC and 40-70 %.
- Air changes (per hr): Filtered fresh air which was passed to atmosphere and not recirculated.
- Photoperiod (hrs dark / hrs light): Artificial lighting, 12 hours light : 12 hours dark

IN-LIFE DATES: From: 18th February 2013 To: 05th May 2013

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
other: 0.1% Tween 80
Details on exposure:
PREPARATION OF DOSING SOLUTIONS:

VEHICLE
- Justification for use and choice of vehicle (if other than water): vehicle assessed in preliminary study and used for other animal testing studies
- Concentration in vehicle: 0.1%
- Amount of vehicle (if gavage): 10 mL/kg
- Lot/batch no. (if required): N/a
- Purity: As supplied
Details on mating procedure:
- M/F ratio per cage: 5/5 (during pre-pairing) and 1/1 (during pairing)

- M/F ratio per cage: 5/5 (during pre-pairing) and 1/1 (during pairing)
- Length of cohabitation: up to two weeks
- Proof of pregnancy: Ejected copulation plugs. sperm in vaginal smear referred to as day 1 of pregnancy
- After successful mating each pregnant female was caged (how): one female per cage
Analytical verification of doses or concentrations:
yes
Duration of treatment / exposure:
4 weeks
Frequency of treatment:
once daily
Details on study schedule:
Duration of treatment
F0 animals
Males - for a minimum of four weeks, including 15 days before pairing.
Females - for 15 days before pairing until Day 6 after birth of the

F1 generation.
F1 animals No direct treatment
Doses / concentrationsopen allclose all
Dose / conc.:
0 mg/kg bw/day (nominal)
Remarks:
Control
Dose / conc.:
60 mg/kg bw/day (nominal)
Dose / conc.:
200 mg/kg bw/day (nominal)
Dose / conc.:
600 mg/kg bw/day (nominal)
No. of animals per sex per dose:
Three groups, each comprising ten male and ten female
Control animals:
yes, concurrent vehicle

Results and discussion

Results: P0 (first parental animals)

General toxicity (P0)

Clinical signs:
no effects observed
Description (incidence and severity):
There were no signs associated with dosing and no general clinical signs related to treatment.
Mortality:
mortality observed, non-treatment-related
Description (incidence):
Treatment up to 600 mg/kg/day was well tolerated, with no adverse effects on general condition or signs related to treatment. One female treated at 600 mg/kg/day was killed for
welfare reasons as it displayed signs of distress, had abdominal distension and was bleeding from the vagina. Macroscopic examination revealed that parturition had been incomplete with
two dead fetuses, two live fetuses and four placentae present in the uterus. This was considered to be the result of an unfortunate normal biological event and not related to
treatment.
Body weight and weight changes:
effects observed, non-treatment-related
Description (incidence and severity):
It was considered that there was no treatment related effect on body weight gain or food consumption on adult males or females during the two-week pre-mating period and no effect
on body weight during gestation or early lactation (to Day 7 post-partum). Males treated at 60, 200 or 600 mg/kg/day had low mean body weight gain during the week after pairing and
females receiving 600 mg/kg/day had marginally low food consumption during the first week of treatment and marginally high food consumption was seen in treated females during
gestation (Days 10-13); however, they were considered not to be toxicologically significant.
Food consumption and compound intake (if feeding study):
effects observed, non-treatment-related
Description (incidence and severity):
It was considered that food consumption was not affected by treatment, during the two week pre-pairing period, or during gestation or lactation.
Food consumption of females treated at 600 mg/kg/day was marginally low (X 0.87 Control) during the first week of treatment and was marginally high in treated females during gestation
(Days 10-13 only), when compared with Control, but these incidences were considered to be un-related to treatment.
Food efficiency:
not specified
Water consumption and compound intake (if drinking water study):
not specified
Ophthalmological findings:
not specified
Haematological findings:
not specified
Clinical biochemistry findings:
not specified
Urinalysis findings:
not specified
Behaviour (functional findings):
not specified
Immunological findings:
not specified
Organ weight findings including organ / body weight ratios:
no effects observed
Description (incidence and severity):
There was no effect of treatment on organ weights.
Gross pathological findings:
effects observed, non-treatment-related
Description (incidence and severity):
Pale areas were seen on the liver of two males treated at 600 mg/kg/day and an enlarged liver was seen in one male treated at 600 mg/kg/day. The nature and incidence of all other findings were consistent with the commonly seen background of macroscopic changes in CD rats at these laboratories.
Neuropathological findings:
not specified
Histopathological findings: non-neoplastic:
no effects observed
Description (incidence and severity):
There were no changes observed in the reproductive organs which were considered to be related to treatment with MTF.
The seminiferous tubules of the testes were evaluated with respect to their stage in the spermatogenic cycle and the integrity of the various cell types present within the different
stages. No cell or stage specific abnormalities were noted. Two males treated at 600 mg/kg/day had pale areas on the liver and the liver of another male
was enlarged. Histopathological investigation revealed centrilobular hypertrophy, which was considered to be an adaptive change resulting from exposure to the test substance.
Histopathological findings: neoplastic:
not specified
Other effects:
not specified

Reproductive function / performance (P0)

Description (incidence and severity):
A higher proportion of females had regular cycles of 4/5 days than expected at 600 mg/kg/day, when compared with Control. The reason for this finding is not known, but as other
investigations for reproductive performance were not affected by treatment, it was considered not to represent an adverse effect of treatment.
Reproductive function: sperm measures:
not specified
Reproductive performance:
no effects observed
Description (incidence and severity):
Pre-coital interval, mating performance and fertility were not affected by treatment. One Control animal (No. 42) was not pregnant and had no sign of any implantations pre and post staining.

Details on results (P0)

There were no changes observed in the reproductive organs which were considered to be related to treatment with MTF. There was considered to be no effect of treatment on pre-coital interval, mating performance, fertility, gestation length, gestation index, sex ratio or survival and development of the offspring, or macroscopic change in the offspring.

Effect levels (P0)

Key result
Dose descriptor:
NOAEL
Effect level:
600 mg/kg bw/day
Based on:
test mat.
Sex:
male/female
Basis for effect level:
clinical signs
body weight and weight gain
food consumption and compound intake
organ weights and organ / body weight ratios
gross pathology
histopathology: non-neoplastic
reproductive function (oestrous cycle)
reproductive performance

Target system / organ toxicity (P0)

Key result
Critical effects observed:
no

Results: F1 generation

General toxicity (F1)

Clinical signs:
effects observed, non-treatment-related
Description (incidence and severity):
Clinical signs recorded for the offspring identified occasional pups with findings, but the type of findings (i.e. cold to touch, bruised muzzle) and incidence were typical of post-natal
offspring and showed no relationship to treatment.
Mortality / viability:
no mortality observed
Description (incidence and severity):
There was no effect of treatment on litter size, offspring survival or sex ratio.
Body weight and weight changes:
no effects observed
Description (incidence and severity):
The slightly higher body weight gain of offspring between Days 1 and 4 of age, following parental treatment at 200 or 600 mg/kg/day, was attributed to the marginally low gains seen
in the Control and low dose group (60 mg/kg/day), which was attributed to the higher than expected mean litter size in these groups.
Food consumption and compound intake (if feeding study):
not specified
Food efficiency:
not specified
Water consumption and compound intake (if drinking water study):
not specified
Ophthalmological findings:
not specified
Haematological findings:
not specified
Clinical biochemistry findings:
not specified
Urinalysis findings:
not specified
Sexual maturation:
not specified
Organ weight findings including organ / body weight ratios:
not specified
Gross pathological findings:
not specified
Histopathological findings:
not specified

Details on results (F1)

There was considered to be no effect of treatment on pre-coital interval, mating performance, fertility, gestation length, gestation index, sex ratio or survival and development of the
offspring, or macroscopic change in the offspring.

Effect levels (F1)

Key result
Dose descriptor:
NOAEL
Generation:
F1
Effect level:
600 mg/kg bw/day (actual dose received)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
viability
clinical signs
body weight and weight gain
gross pathology

Target system / organ toxicity (F1)

Key result
Critical effects observed:
no

Overall reproductive toxicity

Key result
Reproductive effects observed:
no

Applicant's summary and conclusion

Conclusions:
It is concluded that once daily oral administration of MTF to male and female Crl:CD(SD) rats for at least four weeks at doses of 60, 200 or 600 mg/kg/day was well tolerated with no
effect of treatment on reproductive performance, including mating performance, fertility and offspring survival and development up to Day 7 of age.
For reproductive/developmental toxicity the no-observed-adverse-effect-level (NOAEL) was considered to be 600 mg/kg/day.