Registration Dossier

Administrative data

Key value for chemical safety assessment

Effects on fertility

Description of key information

A screening for reproductive / developmental toxicity (OECD 421) is available and well conducted.

Link to relevant study records
Reference
Endpoint:
screening for reproductive / developmental toxicity
Type of information:
experimental study
Adequacy of study:
key study
Study period:
from 2009-05-27 to 2010-04-27
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: see 'Remark'
Remarks:
This study has been performed in compliance with the: Swiss Ordinance relating to Good Laboratory Practice adopted May 18th, 2005 [SR 813.112.1]. This Ordinance is based on the OECD Principles of Good Laboratory Practice, as revised in 1997 and adopted on November 26th, 1997 by decision of the OECD Council [C (97)186/Final]. These principles were compatible with Good Laboratory Practice regulations specified by regulatory authorities throughout the European Community, the United States (EPA and FDA), and Japan (MHLW, MAFF and METI). There were no circumstances that may have affected the quality or integrity of the data.
Reason / purpose:
reference to same study
Qualifier:
according to
Guideline:
OECD Guideline 421 (Reproduction / Developmental Toxicity Screening Test)
Deviations:
no
GLP compliance:
yes (incl. certificate)
Remarks:
TYPES OF QA INSPECTIONS: Study plan; study based (animal delivery, raw data, test system, test item, dose preparation, treatment, body weight, necropsy); process based (analytics work up); analytic appendix; report
Limit test:
no
Species:
rat
Strain:
Wistar
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
Animals: Rat, HanRcc: WIST(SPF)
Rationale: Recognized by international guidelines as a recommended test system.
Breeder: Harlan Laboratories Ltd., Laboratory Animal Services, Wölferstrasse 4, 4414 Füllinsdorf / Switzerland
Number of Animals: 40 males (10 per group) and 40 females (10 per group)
Age (at Start of Treatment): 11 weeks
Body Weight Range (at Start of Treatment): Males (291 to 338 g) and females (185 to 211 g)
Identification: Cage card and individual animal number (ear tattoo).
Randomization: Computer-generated random algorithm. In addition body weights (recorded on the day of allocation) were taken into consideration in order to ensure similar mean body weights in all groups.
Acclimatization: Under test conditions after health examination. Only animals without any visible signs of illness were used for the study.

ENVIRONMENTAL CONDITIONS
Conditions: Standard laboratory conditions. Air-conditioned with 10 - 15 air changes per hour, continuously monitored environmental conditions (temp. range: 22 ± 3 °C; relative humidity range: 30 – 86.7%). There was 12 hour fluorescent light / 12-hour dark cycle with music during the light period.
Accommodation: Individually in Makrolon type-3 cages with wire mesh tops and sterilized standard softwood bedding (‘Lignocel’J. Rettenmaier & Söhne GmbH & CoKG, 73494 Rosenberg / Germany, imported by Provimi Kliba SA, 4303 Kaiseraugst / Switzerland). During the pre-pairing period, cages with males were interspersed amongst those holding females to promote the development of regular estrus cycles.
Diet: Pelleted standard Kliba Nafag 3433 rodent maintenance diet (Provimi Kliba SA, 4303 Kaiseraugst / Switzerland) was available ad libitum (batch no. 15/09).
Water: Community tap-water from Füllinsdorf was available ad libitum in water bottles.
Route of administration:
oral: gavage
Vehicle:
water
Details on exposure:
DOSE FORMULATIONS

The dose formulations were prepared daily using the test item as supplied by the Sponsor.

LCE08125 was weighed into a glass beaker on a tared precision balance and approximately 80% of the vehicle was added (w/v). Using an appropriate homogenizer, a homogeneous suspension was prepared. Having obtained a homogeneous mixture, the remaining vehicle was added. Separate formulations were prepared for each concentration.

Homogeneity of the test item in the vehicle was maintained during the daily administration period using a magnetic stirrer.

STORAGE OF DOSE FORMULATIONS

Dose formulations were stored at room temperature (20 ± 5 °C) in brown glass beakers.

TREATMENT

Method: Oral, by gavage
Rationale for Method: Administration by gavage is a common and accepted route of exposure for studies of this type.
Frequency of Administration: Daily
Target Dose Levels: Group 1: 0 mg/kg/day (control group); Group 2: 100 mg/kg/day; Group 3: 300 mg/kg/day; Group 4: 1000 mg/kg/day
Rationale for Dose Level Selection: The dose levels were selected based on a previous dose range finding toxicity study in Han Wistar, Harlan Laboratories Study C37152, using dose levels of 100, 300 and 1000 mg/kg/day, resulting in a NOEL of 1000 mg/kg/day.
Dose Volume: 10 mL/kg body weight
Duration of Acclimatization Period: 9 days
Duration of Treatment Period: Males (minimum 4 weeks); females (approximately 7 weeks)


Details on mating procedure:
MATING, GESTATION AND LACTATION

During the pairing period, females were housed with sexually mature males (1:1) in special automatic mating cages i.e. with synchronized timing to initiate the nightly mating period, until evidence of copulation was observed. This system reduced the variation in the copulation times of the different females. The females were removed and housed individually if:

- the daily vaginal smear was sperm positive, or
- a copulation plug was observed.

The day of mating was designated day 0 post coitum.

All dams were allowed to give birth and rear their litters (F1 pups) up to day 4 post partum. Day 0 was designated as the day on which a female had delivered all her pups.
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
ANALYSIS OF DOSE FORMULATIONS

On the first treatment day one sample from the control group as well as three samples (top, middle and bottom) of about 2 g of each concentration were taken prior to dosing for analysis of concentration and homogeneity. Samples of about 2 g of each concentration were taken from the middle to confirm stability (4 hrs and 7 days). During the last week of the treatment, samples were taken from the middle to confirm concentration. The aliquots for analysis of dose formulations were frozen (-20 ± 5 °C) and delivered on dry ice to Analytic Department (Harlan Laboratories Ltd., Itingen / Switzerland) and stored there at -20 ± 5 °C until analysis.

The samples were analyzed by HPLC coupled to an ELSD detector following an analytical procedure developed at Harlan Laboratories. The test item was used as the analytical standard. Analyzed samples were not discarded without written consent from the study director.

The application formulations investigated during the study were found to comprise LCE08125 in the range of 85.2% to 101.7% and, thus, the required content limit of ±20% with reference to the nominal concentration was met. The homogeneous distribution of LCE08125 in the preparations was approved because single results found did not deviate more than 1.9% (<15%) from the corresponding mean. In addition, the test item was found to be stable in application formulations when kept 7 days at room temperature due to recoveries which met the variation limit of 10% from the time-zero (homogeneity) mean.
Duration of treatment / exposure:
Males: Minimum 4 weeks
Females: Approximately 7 weeks
Frequency of treatment:
Daily
Details on study schedule:
Acclimatization: 9 days (males and females)
First Test Item Administration: Day 1 of pre-pairing (males and females)
Pre-Pairing: 14 days (males and females)
Pairing: 14 days maximum (males and females)
Gestation: Approximately 21 days (females)
Treatment Ends: On day 3 post partum (females); on day before sacrifice (males)
Necropsy: On day 4 post partum (females); after a minimum of 28 days treatment (males)
Remarks:
Doses / Concentrations:
100 mg/kg bw/day
Basis: actual ingested
Remarks:
Doses / Concentrations:
300 mg/kg bw/day
Basis: actual ingested
Remarks:
Doses / Concentrations:
1000 mg/kg bw/day
Basis: actual ingested
No. of animals per sex per dose:
10
Control animals:
yes, concurrent vehicle
Parental animals: Observations and examinations:
Viability / Mortality: Twice daily
Clinical Signs: Daily cage-side clinical observations (once daily, during acclimatization and up to day of necropsy). Additionally females were observed for signs of difficult or prolonged parturition, and behavioral abnormalities in nesting and nursing.
Food Consumption: Males (weekly during pre-pairing and after pairing periods); females (pre-pairing period days 1 - 8 and 8 - 14, gestation period days 0 - 7, 7 - 14 and 14 - 21 post coitum and lactation period days 1 - 4 post partum). No food consumption was recorded during the pairing period.
Body Weights: Recorded daily from treatment start to day of necropsy.
Litter observations:
The litters were examined for litter size, live births, still births and any gross anomalies. The sex ratio of the pups was recorded. Pups were weighed individually (without identification) on days 0 (if possible), 1 and 4 post partum.
Postmortem examinations (parental animals):
TERMINATION OF THE STUDY

Males were sacrificed after they had been treated for at least 28 days. Dams were sacrificed on day 4 post partum.

When birth did not occur on the expected date (day 21 post coitum), the dams were sacrificed on day 25 post coitum.

NECROPSY

All parent animals were sacrificed by an injection of sodium pentobarbital. All P generation animals were exsanguinated.

All parent animals were examined macroscopically for any structural changes, either at the scheduled necropsy or during the study if death occurred.

For the parent animals, special attention was directed at the organs of the reproductive system.
The number of implantation sites and corpora lutea was recorded for all dams with litters. The uteri of non-pregnant parent females were placed in a solution of ammonium sulfide to visualize possible hemorrhagic areas of implantation sites.

ORGAN WEIGHTS

The testes and epididymides of all parental males were weighed as pairs.

TISSUE PRESERVATION

The ovaries from all parental females were preserved in neutral phosphate buffered 4% formaldehyde solution.

The testes and epididymides from all parental males were preserved in Bouin’s fixative. The prostate and seminal vesicles from all parental males were fixed in neutral phosphate buffered 4% formaldehyde solution.

HISTOTECHNIQUE

All organ and tissue samples to be examined by the principal investigator were processed, embedded and cut at an approximate thickness of 2 - 4 micrometers and stained with hematoxylin and eosin. Additionally, the testis was stained by PAS–hematoxylin. Special stains were used at the discretion of the study pathologist.

HISTOPATHOLOGY

Slides of all organs and tissues listed collected at terminal sacrifice from the animals of the control and high-dose groups were examined by the principal investigator. The same applied to all occurring gross lesions and to all animals which died spontaneously or had to be terminated in extremis.

Special emphasis was made on the stages of spermatogenesis and histopathology of interstitial cell structure.

Histological examination of ovaries was carried out on females that did not give birth. In addition, microscopic examination of the reproductive organs of all infertile males was made, if necessary.

Postmortem examinations (offspring):
TERMINATION OF THE STUDY

Pups were sacrificed on day 4 post partum.

NECROPSY

All pups were sacrificed by an injection of sodium pentobarbital.

All pups were examined macroscopically for any structural changes, either at the scheduled necropsy or during the study if death occurred.
Statistics:
The following statistical methods were used to analyze food consumption, body weights and reproduction data:
- Means and standard deviations of various data were calculated.
- The Dunnett-test (many to one t-test) based on a pooled variance estimate was applied if the variables could be assumed to follow a normal distribution for the comparison of the treated groups and the control groups for each sex.
- The Steel-test (many-one rank test) was applied instead of the Dunnett-test when the data could not be assumed to follow a normal distribution.
- Fisher's exact-test was applied if the variables could be dichotomized without loss of information.
Reproductive indices:
From the on-line recorded reproduction data, the following parameters were calculated: fertility indices, mean precoital time, post-implantation losses, mean litter size.
Offspring viability indices:
From the on-line recorded reproduction data, the following parameters were calculated: dead/live pups at first litter check, pup sex ratios and postnatal loss (up to day 4 post partum).
Clinical signs:
no effects observed
Mortality:
no mortality observed
Body weight and weight changes:
effects observed, non-treatment-related
Description (incidence and severity):
Body Weights of Males
No test item-related changes were observed in mean body weight and mean body weight gain in any groups during the entire duration of the study. In higher group, the statistically significantly higher body weight gain on days 6 and 8 of the pairing period was considered to be incidental.
Body Weights of Females
Mean body weight and mean body weight gain were not affected by treatment with the test item in all groups during the entire duration of the period. In absence of any obvious dose-relationship, the statistically significantly higher body weight gain in groups 2 and 3 (i.e.100 mg/kg body weight/day and 300 mg/kg body weight/day) noted during the prepairing period was considered to be incidental.
Food consumption and compound intake (if feeding study):
effects observed, non-treatment-related
Description (incidence and severity):
In male groups mean food consumption was not affected by the treatment with the test item
In females at the high dose level, mean food consumption was slightly slightly lower during the first week of the prepairing period (-7.1% compared to the control), and during the lactation period (-8.4%). None of these reductions was statistically significant and they were not considered of adverse nature since they did not affect mean body weight gain or were transient..
Food efficiency:
no effects observed
Water consumption and compound intake (if drinking water study):
not examined
Ophthalmological findings:
not examined
Haematological findings:
not examined
Clinical biochemistry findings:
not examined
Urinalysis findings:
not examined
Behaviour (functional findings):
not examined
Immunological findings:
not examined
Organ weight findings including organ / body weight ratios:
no effects observed
Gross pathological findings:
effects observed, non-treatment-related
Description (incidence and severity):
During necropsy of parent animals type and incidence of abnormal findings did not give any
indication of a test item-related effect.
Males
One male in higher treated group had a red brown diaphragmatic hernia. One male each in group 2 and 3 had reduced size of both testes and epididymides. These findings were considered incidental.
Females
No abnormal findings were observed during the macroscopical examination of females.
Neuropathological findings:
not examined
Histopathological findings: non-neoplastic:
effects observed, non-treatment-related
Description (incidence and severity):
The histopathological evaluation of the reproductive organs did not reveal any relevant changes in the high-dose animals.
Special emphasis on stages of spermatogenesis and histopathology of interstitial testicular cell structure did not reveal any differences between control (group 1) and high-dose (group 4) males.
Group 2 male No. 20 and group 3 male No. 29 both showed bilaterally a reduced size of testes and epididymides. These gross lesions correlated histopathologically in both males with marked diffuse tubular degeneration. In male No. 20, the atrophy was accompanied by moderate multifocal spermatic giant cells and minimal diffuse edema, and any stages of spermatogenesis could be recognized anymore. In male No. 29, the atrophy was accompanied by slight moderate multifocal spermatic giant cells and slight diffuse edema, and only in single tubules the stages of
spermatogenesis could unilaterally be recognized (stages IV-V, VIII, and III-XIV). In epididymis of male No. 20, a marked oligospermia with moderate intratubular cellular debris was present and in male No. 29, a massive oligospermia with moderate intratubular cellular
debris and unilateral focal lymphoid cell infiltration was noted. All these findings were considered to be most likely incidental and unrelated to the test item. The prostate and seminal vesicles did not reveal any morphological changes.
The ovaries from group 2 female No. 60 and group 3 female No. 69 which were both not pregnant did not show any morphological changes. The reason for missing pregnancy was the infertility present in the according mating males Nos. 20 and 29.
Histopathological findings: neoplastic:
no effects observed
Other effects:
not examined
Reproductive function: oestrous cycle:
not examined
Reproductive function: sperm measures:
no effects observed
Reproductive performance:
no effects observed
1 IN-LIVE DATA - PARENTAL ANIMALS

1.1 CLINICAL SIGNS OR OBSERVATIONS

All animals survived until the scheduled necropsy. No clinical signs were observed during the study.

1.2 FOOD CONSUMPTION OF MALES

Mean food consumption was not affected by the treatment with the test item. Lower food consumption which occurred in groups 2 and 3 during the first week of pre-pairing and during the after pairing period was considered to be incidental since there was no dose-dependent pattern.

See attached figures and tables on pp. 2 to 3; 22 to 23; 27 to 28

1.3 FOOD CONSUMPTION OF FEMALES

In group 4, mean food consumption was slightly lower during the first week of the pre-pairing period (-7.1% compared to the control), and during the lactation period (-8.4%). None of these reductions was statistically significant and they were not considered of adverse nature since they did not affect mean body weight gain or were transient.

In groups 2 and 3, no effects were noted on food consumption during the pre-pairing, gestation and lactation periods.

See attached figures and tables on pp. 4 to 6; 24 to 26; 29 to 31

1.4 BODY WEIGHTS OF MALES

No test item-related changes were observed in mean body weight and mean body weight gain in any groups during the entire duration of the study. In group 4, the statistically significantly higher body weight gain on days 6 and 8 of the pairing period was considered to be incidental.

See attached figures and tables on pp. 7 to 9; 14 to 16; 32 to 34; 40 to 42; 48 to 50

1.5 BODY WEIGHTS OF FEMALES

Mean body weight and mean body weight gain were not affected by treatment with the test item in all groups during the entire duration of the period. In absence of any obvious dose-relationship, the statistically significantly higher body weight gain in groups 2 and 3 noted during the pre-pairing period was considered to be incidental.

See attached figures and tables on pp. 10 to 13; 17 to 20; 35 to 39; 43 to 47; 51 to 53


2 REPRODUCTION AND BREEDING DATA

2.1 MATING PERFORMANCE AND FERTILITY

The median and mean precoital times were unaffected by treatment with the test item. Mean precoital times were 3.4, 3.7, 2.4 and 2.2 days in order of ascending dose level. The median precoital time was 4, 4, 2 and 2 days in order of ascending dose level.

One female (no. 60) in group 2 and one female (no. 69) in group 3 were not pregnant. Thus the fertility indices were 100.0%, 90.0%, 90.0% and 100.0% in groups 1, 2, 3 and 4.

2.2 DURATION OF GESTATION

The mean duration of gestation was unaffected by treatment with the test item. Mean duration of gestation was 21.2, 21.4, 21.3 and 21.4 days, in order of ascending dose level.

2.3 CORPORA LUTEA COUNT

The mean number of corpora lutea per dam (determined at necropsy) was similar in all groups (13.8, 14.1, 14.6 and 13.7 in order of ascending dose level) and gave no indication of a test item-related effect.

2.4 IMPLANTATION RATE AND POST-IMPLANTATION LOSS

The mean number of implantations per dam and the post-implantation losses were unaffected by treatment with the test item.

The mean numbers of implantations per litter were 13.6, 12.9, 13.2 and 12.4 in order of ascending dose level. The slightly lower number of implantation sites in group 4 was due to one female (no. 76) which had only six implantation sites. The mean incidence of post-implantation loss as a percentage of total implantations was 5.1, 6.9, 5.0 and 5.6% in order of ascending dose level.

2.5 LITTER SIZE AT FIRST LITTER CHECK

The number of live pups at first litter check was unaffected by treatment with the test item. The mean number of live pups per litter was 12.9, 12.0, 12.6 and 11.7 in order of ascending dose level.

2.6 POSTNATAL LOSS DAYS 0 - 4 POST PARTUM

Postnatal loss up to day 4 post partum was not affected by the treatment with the test item.

The viability indices were 100.0%, 100.0%, 99.1% and 99.1% in order of ascending dose levels.


3 TERMINAL FINDINGS - PARENTAL ANIMALS

3.1 ORGAN WEIGHTS

In males, weights (absolute and relative to body weight) of testes and epididymides were not affected by the treatment with the test item in any groups.

3.2 MACROSCOPICAL FINDINGS

During necropsy of parent animals type and incidence of abnormal findings did not give any indication of a test item-related effect.

Males: One male in group 4 had a red brown diaphragmatic hernia. One male each in group 2 and 3 had reduced size of both testes and epididymides. These findings were considered incidental.

Females: No abnormal findings were observed during the macroscopical examination of females.

3.3 HISTOPATHOLOGY FINDINGS

The histopathological evaluation of the reproductive organs did not reveal any relevant changes in the high-dose animals.
Special emphasis on stages of spermatogenesis and histopathology of interstitial testicular cell structure did not reveal any differences between control (group 1) and high-dose (group 4) males.

Group 2 male No. 20 and group 3 male No. 29 both showed bilaterally a reduced size of testes and epididymides. These gross lesions correlated histopathologically in both males with marked diffuse tubular degeneration. In male No. 20, the atrophy was accompanied by moderate multifocal spermatic giant cells and minimal diffuse edema, and any stages of spermatogenesis could be recognized anymore. In male No. 29, the atrophy was accompanied by slight moderate multifocal spermatic giant cells and slight diffuse edema, and only in single tubules the stages of spermatogenesis could unilaterally be recognized (stages IV-V, VIII, and III-XIV). In epididymis of male No. 20, a marked oligospermia with moderate intratubular cellular debris was present and in male No. 29, a massive oligospermia with moderate intratubular cellular debris and unilateral focal lymphoid cell infiltration was noted. All these findings were considered to be most likely incidental and unrelated to the test item. The prostate and seminal vesicles did not reveal any morphological changes.
The ovaries from group 2 female No. 60 and group 3 female No. 69 which were both not pregnant did not show any morphological changes. The reason for missing pregnancy was the infertility present in the according mating males Nos. 20 and 29.
Key result
Dose descriptor:
NOAEL
Effect level:
1 000 mg/kg bw/day (actual dose received)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: Since no adverse effects were observed, the general NOAEL (No Observed Adverse Effect Level) was established at 1000 mg/kg/day. NOAL = highest dose tested
Key result
Dose descriptor:
NOEL
Effect level:
1 000 mg/kg bw/day (actual dose received)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: Under the conditions of this study, the NOEL (No Observed Effect Level) for reproduction/ developmental toxicity was considered to be 1000 mg/kg/day. NOEL = highest dose tested
Remarks on result:
other: Generation: reproduction/development (migrated information)
Clinical signs:
not examined
Mortality:
not examined
Body weight and weight changes:
not examined
Food consumption and compound intake (if feeding study):
not examined
Food efficiency:
not examined
Water consumption and compound intake (if drinking water study):
not examined
Ophthalmological findings:
not examined
Haematological findings:
not examined
Clinical biochemistry findings:
not examined
Urinalysis findings:
not examined
Behaviour (functional findings):
not examined
Immunological findings:
not examined
Organ weight findings including organ / body weight ratios:
not examined
Gross pathological findings:
not examined
Neuropathological findings:
not examined
Histopathological findings: non-neoplastic:
not examined
Histopathological findings: neoplastic:
not examined
Reproductive function: oestrous cycle:
not examined
Reproductive function: sperm measures:
not examined
Reproductive performance:
not examined
Remarks on result:
not measured/tested
Remarks:
no second parental generation
Clinical signs:
no effects observed
Description (incidence and severity):
At first litter check and during lactation no findings were observed during the external examination of pups.
No clinical signs were noted in any groups.
Mortality / viability:
no mortality observed
Body weight and weight changes:
no effects observed
Food consumption and compound intake (if feeding study):
not examined
Food efficiency:
not examined
Water consumption and compound intake (if drinking water study):
not examined
Ophthalmological findings:
not examined
Haematological findings:
not examined
Clinical biochemistry findings:
not examined
Urinalysis findings:
not examined
Sexual maturation:
not examined
Organ weight findings including organ / body weight ratios:
not examined
Gross pathological findings:
no effects observed
Histopathological findings:
not examined
Other effects:
no effects observed
Description (incidence and severity):
Sex ratios at first litter check and on day 4 post partum were unaffected by exposure to the test
item.
Behaviour (functional findings):
not examined
Developmental immunotoxicity:
not examined
1 LITTER DATA - F1 PUPS

1.1 EXTERNAL EXAMINATION AT FIRST LITTER CHECK AND DURING LACTATION

At first litter check and during lactation no findings were observed during the external examination of pups.

No clinical signs were noted in any groups.

1.2 SEX RATIOS

Sex ratios at first litter check and on day 4 post partum were unaffected by exposure to the test item.

The proportion of males on day 4 post partum was 47%, 42%, 49% and 46% in order of ascending dose level.

1.3 PUP WEIGHTS TO DAY 4 POST PARTUM

Mean pup weights were unaffected by treatment with the test item. On day 1 post partum mean pup weights were 5.9, 6.0, 6.1 and 6.3 g in order of ascending dose level.

Mean pup weight gain during lactation was also not affected by the treatment with the test item. Mean pup weights on day 4 post partum were 8.6, 8.8, 8.9 and 9.1 g in order of ascending dose level.

1.4 MACROSCOPICAL FINDINGS

No abnormal findings were noted at macroscopic examination of the pups.

Key result
Dose descriptor:
NOAEL
Generation:
F1
Effect level:
1 000 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: no significant adverse effect noted
Clinical signs:
not examined
Mortality / viability:
not examined
Body weight and weight changes:
not examined
Food consumption and compound intake (if feeding study):
not examined
Food efficiency:
not examined
Ophthalmological findings:
not examined
Haematological findings:
not examined
Clinical biochemistry findings:
not examined
Urinalysis findings:
not examined
Sexual maturation:
not examined
Organ weight findings including organ / body weight ratios:
not examined
Gross pathological findings:
not examined
Histopathological findings:
not examined
Behaviour (functional findings):
not examined
Developmental immunotoxicity:
not examined
Remarks on result:
not measured/tested
Remarks:
only F1 generation examined
Reproductive effects observed:
not specified

SUMMARY OF PERFORMANCE

P Animals Breeding for F1 Litters

Group
(mg/kg/day)

1
(0)

2
(100)

3
(300)

4
(1000)

Female numbers

41-50

51-60

61-70

71-80

Number of females paired

10

10

10

10

Number of females mated

10

10

10

10

Number of pregnant females (A)

10

9

9

10

Number of females which reared their pups until day 4 post partum

10

9

9

10

(A) Female Nos. 60 and 69 were not pregnant.

 

Conclusions:
This study is a valid investigation of the toxicological effects resulting from repeated oral-gavage administration of the test item LCE08125 to rats. LCE08125 was administered in Milli-Q-Water as vehicle at dosages of 100, 300, and 1000 mg/kg body weight/day, and controls received the vehicle only. LCE08125 was administered to male rats for at least 28 days and to female rats for 14 days prior to pairing, through the pairing and gestation periods until the F1 generation reached day 3 post partum.

Since no adverse effects were observed, the general NOAEL (No Observed Adverse Effect Level) was established at 1000 mg/kg/day.

Under the conditions of this study, the NOEL (No Observed Effect Level) for reproduction/ developmental toxicity was considered to be 1000 mg/kg/day.
Executive summary:

The purpose of this study was to generate preliminary information concerning the effects of LCE08125 on male and female reproductive performance such as gonadal function, mating behavior, conception and parturition.

Four groups of 10 males and 10 females were treated by gavage with LCE08125 once daily. Males were treated over a 14-day pre-pairing period and during the pairing period up to one day before necropsy. Females were treated throughout the pre-pairing, pairing, gestation and lactation period up to day 3 post partum.

The following dose levels were used:

Group 1:                0 mg/kg body weight/day (control group)

Group 2             100 mg/kg body weight/day

Group 3:            300 mg/kg body weight/day

Group 4           1000 mg/kg body weight/day

 

A standard dose volume of 10 mL/kg body weight with a daily adjustment to the actual body weight was used. Control animals were dosed with the vehicle alone (highly purified water).

  

PARENTAL ANIMALS

 

General Tolerability

All animals survived until the scheduled necropsy and no clinical signs were observed during the entire study.

 

Food Consumption and Body Weights

In males and females, mean food consumption, mean body weight and mean body weight gains were not adversely affected by the treatment with the test item for the whole duration of the treatment.

 

Reproductive Data

Relevant reproduction data (mean number of corpora lutea, mean number of implantations per dam, and the post-implantation losses) were not affected by treatment with the test item. The mean duration of gestation was similar in all groups.

 

Organ Weights

Mean weight of testes and epididymides were not affected by treatment with the test item.

 

Macroscopical Findings and Histopathological Examinations

No test item-related effects were noted in reproductive organs in high-dose animals.

 

LITTER DATA - F1 PUPS

Mean litter size at first litter check and on day 4 post partum was not affected by treatment with the test item. Mean pup weights and weight gains were also not affected.

Effect on fertility: via oral route
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
NOAEL
1 000 mg/kg bw/day
Study duration:
subacute
Species:
rat
Quality of whole database:
The key study was conducted according to the OECD test method 421.
Effect on fertility: via inhalation route
Endpoint conclusion:
no study available
Effect on fertility: via dermal route
Endpoint conclusion:
no study available
Additional information

Short description of key information:
An OECD test n°421 was conducted on rats with the test substance, NOAEL parental and reproduction = 1000 mg/kg bw/d

Justification for selection of Effect on fertility via oral route:
A well conducted reproduction toxicity study is available

Justification for selection of Effect on fertility via inhalation route:
Based on exposure considerations, this endpoint is not relevant for the test item.
This substance is not volatile (vapor pressure of 3.6 10-4 Pa at 25°C).
No exposition is expected via this route.

Justification for selection of Effect on fertility via dermal route:
According to ECHA guidance - Chapter R7C, if water solubility is above 10000 mg/l and the log Kow value below 0 the substance may be too hydrophilic to cross the lipid rich environment of the stratum corneum. Dermal uptake for these substances will be low.
So it is more relevant to take into considerations the oral repeated toxicity study.

Effects on developmental toxicity

Description of key information

Two developmental toxicity studies (OECD 414) are available on supporting "source substances" (read-across: CAS 110615-47-9 and xylitol) and showed no adverse effect (NOAEL for maternal and developmental toxicity = 1000 mg/kg.

Based on robust developmental toxicity studies available on the source substances, the NOAEL of the target substance for the developmental toxicity (OECD 414, oral, rat) can be determined at 1000 mg/kg bw by read-across.

Link to relevant study records
Reference
Endpoint:
developmental toxicity
Type of information:
read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
weight of evidence
Reliability:
2 (reliable with restrictions)
Justification for type of information:
JUSTIFICATION FOR DATA WAIVING
See the document in section 13 of IUCLID entitled "Rationale & justification for the read-across for “D-glucopyranose, oligomers, xylityl glycosides and 1,4 anhydro D-xylitol and D-xylitol” (Human health endpoints)"
Reason / purpose:
read-across source
Reason / purpose:
read-across source
Qualifier:
equivalent or similar to
Guideline:
OECD Guideline 414 (Prenatal Developmental Toxicity Study)
Species:
rat
Dose descriptor:
NOAEL
Effect level:
1 000 mg/kg bw/day
Basis for effect level:
other: no adverse effect noted
Dose descriptor:
NOAEL
Effect level:
1 000 mg/kg bw/day (nominal)
Basis for effect level:
other: no adverse effect noted
Abnormalities:
not specified
Developmental effects observed:
not specified
Conclusions:
Based on robust developmental toxicity studies available on the source substances, the NOAEL of the target substance for the developmental toxicity (OECD 414, oral, rat) can be determined at 1000 mg/kg bw by read-across.
See the document in section 13 of IUCLID entitled "Rationale & justification for the read-across for “D-glucopyranose, oligomers, xylityl glycosides and 1,4 anhydro D-xylitol and D-xylitol” (Human health endpoints)"
Effect on developmental toxicity: via oral route
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
NOAEL
1 000 mg/kg bw/day
Quality of whole database:
All the studies used to complete the read-across approach for this endpoint were robust studies (Klimisch 1 or 2)
Effect on developmental toxicity: via inhalation route
Endpoint conclusion:
no study available
Effect on developmental toxicity: via dermal route
Endpoint conclusion:
no study available
Additional information

Justification for selection of Effect on developmental toxicity: via oral route:
The NOAEL value was determined based on a read-across approach (see section 13 of IUCLID)

Justification for selection of Effect on developmental toxicity: via inhalation route:
Based on exposure considerations, this endpoint is not relevant for the test item.
This substance is not volatile (vapor pressure of 3.6 10-4 Pa at 25°C).
No exposition is expected via this route.

Justification for selection of Effect on developmental toxicity: via dermal route:
According to ECHA guidance - Chapter R7C, if water solubility is above 10000 mg/l and the log Kow value below 0 the substance may be too hydrophilic to cross the lipid rich environment of the stratum corneum. Dermal uptake for these substances will be low.
So it is more relevant to take into considerations the oral repeated toxicity study.

Justification for classification or non-classification

According to toxicity to reproduction studies and developmental toxicity studies available, the registered substance is not classified according to CLP regulation.