Registration Dossier

Administrative data

Key value for chemical safety assessment

Effects on fertility

Link to relevant study records
Reference
Endpoint:
screening for reproductive / developmental toxicity
Remarks:
based on test type (migrated information)
Type of information:
experimental study
Adequacy of study:
key study
Study period:
no data
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: 1a - OECD Guideline GLP-compliant study.
Qualifier:
according to
Guideline:
OECD Guideline 421 (Reproduction / Developmental Toxicity Screening Test)
Deviations:
yes
Remarks:
The relative humidity recorded in the animal room was sometimes out of the target range specified in the study plan.
GLP compliance:
yes (incl. certificate)
Limit test:
no
Species:
rat
Strain:
Sprague-Dawley
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Janvier, Le Genest-Sainte-Isle, France.
- Age at study initiation: (P) x 10 wks
- Weight at study initiation: (P) Males: 403-458 g; Females: 253-284 g
- Fasting period before study: no
- Housing: individually, except during prairing in wire-mesh cages (43.0. x 21.5 x 18.0 cm)
- Diet : ad libitum, A04 C P2.5 powder rodent maintenance diet, batch No. 71219 (SAFE, Augy, France)
- Water : ad libitum, tap water (filtered with a 0.22 µm filter)
- Acclimation period: 8 days


ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 +/-2
- Humidity (%): 50 +/- 20
- Air changes (per hr): about 12 cycles/hour of filtered, non-recycled air
- Photoperiod : 12 hrs dark / 12 hrs light (7:00-19:00)


IN-LIFE DATES: From: To:
Route of administration:
oral: feed
Vehicle:
unchanged (no vehicle)
Details on exposure:
PREPARATION OF DOSING SOLUTIONS: For each concentration, a premix containing the total amount of test item was prepared in the diet using a mixer. The final mix was prepared by dispersing the premix in the remaining diet in the mixer and then mixing for at least 10 minutes.


DIET PREPARATION
- Rate of preparation of diet (frequency): The dietary admixtures were prepared on a weekly basis in weeks 1 and 2 then two or three times a week from week 3 by the CIT Pharmacy.
- Mixing appropriate amounts with (Type of food): rodent diet A04 C P2.5, batch No. 71219.
- Storage temperature of food: in closed bags at room temperature prior to use


Details on mating procedure:
- M/F ratio per cage: 1/1
- Length of cohabitation: each female was placed with the same male until mating ocurred or 14 days had elapsed.
- Proof of pregnancy: vaginal plug or sperm in vaginal smearreferred to as day 0 of pregnancy
- After 14 days of unsuccessful pairing replacement of first male by another male with proven fertility.
- Further matings after two unsuccessful attempts: no
- After successful mating each pregnant female was caged (how): individually
- Any other deviations from standard protocol: no
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
The concentration of the test item was determined in samples of each control and test item dosage form prepared for use during weeks 1, 4 and 6. On each day of analysis, duplicate samples were taken at three different levels (top, middle and bottom) of the mixer (groups 2, 3 and 4) and a single level of the container (control group) for determination of the test item concentration.
Duration of treatment / exposure:
in the males:
- 2 weeks before pairing,
- during the pairing period (3 weeks),
- until sacrifice (at least 5 weeks in total),

in the females:
- 2 weeks before pairing,
- during the pairing period (3 weeks),
- during gestation,
- during lactation until day 4 p.p. inclusive (or until sacrifice),
- until sacrifice for non-pregnant females.
Frequency of treatment:
continuous (in diet)
Remarks:
Doses / Concentrations:
1250, 5000 and 20000 ppm
Basis:
nominal in diet
No. of animals per sex per dose:
10
Control animals:
yes, plain diet
Details on study design:
- Dose selection rationale: The dose-levels were selected in agreement with the sponsor, following the results of a previous 4-week toxicity study.
Positive control:
No
Parental animals: Observations and examinations:
CAGE SIDE OBSERVATIONS: Yes
- Time schedule: Each animal was checked for mortality or signs of morbidity once a day before the treatment period and twice a day during the treatment period including weekend and public holidays.



DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: From arrival, the animals were observed once a day as part of routine examinations. From the start of treatment period, each animal was observed once a day, at approximately the same time for the recording of clinical signs.


BODY WEIGHT: Yes
- Time schedule for examinations: The body weight of each male was recorded on the first day of treatment (day 1), then once a week until sacrifice. The body weight of each female was recorded on the first day of treatment (day 1), then once a week until mated (or until sacrifice) and on days 0, 7, 14 and 20 postcoitum (p.c.) and days 1 and 5 post-partum (p.p.).


FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study):
- Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: Yes




Oestrous cyclicity (parental animals):
The estrous cycle stage was determined from a fresh vaginal lavage (stained with methylene blue) each morning during the pairing period until the females had mated.
Sperm parameters (parental animals):
Parameters examined in P male parental generations:
testis weight, epididymis weight.
Litter observations:
STANDARDISATION OF LITTERS
- Performed on day 4 postpartum: no


PARAMETERS EXAMINED
The following parameters were examined in F1 offspring:
number and sex of pups, stillbirths, live births, postnatal mortality, presence of gross anomalies, weight gain, physical or behavioural abnormalities.


GROSS EXAMINATION OF DEAD PUPS:
yes, for external and internal abnormalities; possible cause of death was not determined for pups born or found dead.
Postmortem examinations (parental animals):
SACRIFICE
- Male animals: All surviving animals after the end of the mating period (at least 5 weeks of treatment in total).
- Maternal animals: All surviving animals on day 5 p.p.


GROSS NECROPSY
- Gross necropsy consisted of external and internal examinations including the cervical, thoracic, and abdominal viscera.


HISTOPATHOLOGY / ORGAN WEIGHTS
The tissues indicated in Table 1 were prepared for microscopic examination and weighed, respectively.
Postmortem examinations (offspring):
SACRIFICE
- The F1 offspring were sacrificed at 5 days of age.
- These animals were subjected to postmortem examinations (macroscopic and/or microscopic examination) as follows: Pups were carefully examined externally for gross external abnormalities and a macroscopic examination was performed.


GROSS NECROPSY
- Gross necropsy consisted of external examinations.


HISTOPATHOLOGY / ORGAN WEIGTHS
No tissues were preserved and organ weights were not examined.
Statistics:
PathData (version 6.2b5) was used for the statistical analysis of organ weight data (level of significance: 0.05 or 0.01). The other data are compared by one-way analysis of variances and Dunnett test (mean values being considered as normally distributed, variances being considered as homogenous) or by fisher exact probability test (proportions).
Reproductive indices:
The following parameters were calculated:
- pre-implantation loss: (number of corpora lutea - number of implantation sites)/ (number of corpora lutea) x 100
- post-implantation loss (manually calculated): (number of implantation sites - number of live concepti) / (number of implantations)x 100
- mating index: (number of mated animals) / (number of paired animals) x 100
- fertility index: (number of pregnant female partners)/ (number of mated pairs) x 100
- gestation index: (number of females with live born pups) / (number of pregnant females) x 100
Offspring viability indices:
The following parameters were calculated:
- live birth index: (number of live born pups)/ (number of delivered pups) x 100
- viability index on day 4 p.p.: (number of surviving pups on day 4 p.p.) / (number of live born pups) x 100
Clinical signs:
effects observed, treatment-related
Body weight and weight changes:
effects observed, treatment-related
Food consumption and compound intake (if feeding study):
effects observed, treatment-related
Organ weight findings including organ / body weight ratios:
no effects observed
Gross pathological findings:
effects observed, treatment-related
Histopathological findings: non-neoplastic:
effects observed, treatment-related
Other effects:
not specified
Reproductive function: oestrous cycle:
no effects observed
Reproductive function: sperm measures:
not examined
Reproductive performance:
no effects observed
CLINICAL SIGNS AND MORTALITY (PARENTAL ANIMALS)
There was no premature deaths that could be related to treatment in the groups treated at 1250, 5000, or 20000 ppm. One male treated at 20000 ppm had abnormal growth of teeth from day 8 and was emaciated during the last week of the study. This observation coincided with low body weight gain recorded from the start of the dosing period and with body weight loss between days 29 and 36 (-5%).

BODY WEIGHT AND FOOD CONSUMPTION (PARENTAL ANIMALS)
The group mean body weight gain of males treated at 20000 ppm was affected. Differences were considerable (-34 % over the whole treatment period) and statistically significant when compared to the mean control values. This effect was observed from the beginning of the study and led to statistically significant differences in mean body weight from day 8. At the end of the pre-mating period (day 15), the body weight of males was 8% lower than controls (p<0.01).
The group mean body weight gain of females treated at 20000 ppm was 56% lower than controls during the premating period and mean body weight was 6% lower than controls (p<0.01) on days 8 and 15. During the first week of gestation, the mean body weight gain of females treated at 20000 ppm was 33% lower than controls (and statistically significant, p<0.01). This initial effect was responsible for the lower global body weight gain (-14%) recorded during this phase. Although this difference was not statistically significant at the end of gestation, it was considered to be treatment-related since the body weight was between 6 and 8 % lower than controls on days 7, 14 and 20 of gestation. The group mean body weight recorded on day 5 of lactation was lower than controls (-13%: p<0.001) as was the mean body weight gain from days 1 to 5 of lactation (32%).
At 5000 ppm, the group mean body weight recorded on day 5 of lactation was 6% lower than controls and statistical significant (p<0.01). The differences in the group mean body weight gain of males treated at 5000 ppm and males and females treated at 1250 ppm were considered not to be of biological importance as they were not dose-related.
(See Table 1 for succinct view of the data in the field "remarks on results including tables and figures")

Food consumption of males and females treated at 20000 ppm was 19 to 26% lower than controls during the premating period (p<0.001). During gestation, the females were still affected and food intake was 17 to 23 % lower than controls (with p values<0.01 or <0.001). Between days 1 and 5 of lactation, the food consumption of females treated at the highest concentration was 20 % lower than the control group (p<0.05). At 5000 ppm, this parameter was affected (or altered) by treatment in females during the first week of the premating period (p<0.05) and during gestation and lactation. Differences were slight (-6% to -14%) and did not correlate well with body weight gain. No effect was observed on male mean food consumption at 5000 or 1250 ppm.

TEST SUBSTANCE INTAKE (PARENTAL ANIMALS)
The mean achieved dosages increased in a near dose-proportionnal manner, except for males and females treated at 20000 ppm during premating. (See Table 2 for succinct view of the data in the field "remarks on results including tables and figures")

REPRODUCTIVE FUNCTION: ESTROUS CYCLE (PARENTAL ANIMALS)
No effect was observed on the estrous cycle

REPRODUCTIVE FUNCTION: SPERM MEASURES (PARENTAL ANIMALS)
not performed

REPRODUCTIVE PERFORMANCE (PARENTAL ANIMALS)
There were 0, 3, 2 and 0 non-pregnant females in the groups treated at 0, 1250, 5000 or 20000 ppm, respectively. The incidence of non-pregnant females was not dose-related and was therefore considered not to have been affected by treatment with the test item. Thus, the fertility index was considered not to be affected by treatment with the test item.

ORGAN WEIGHTS (PARENTAL ANIMALS)
No differences were noted between treated and control males in the absolute and relative testis or epididymis weights.

GROSS PATHOLOGY (PARENTAL ANIMALS)
Renal test item-related findings were noted in a dose-related manner, i.e. there were dilated pelvis in 2/10 males given 5000 ppm test item and in 4/10 males given 20000 ppm test item, and granular kidneys with green content in 1/10 females given 20000 ppm test item.

HISTOPATHOLOGY (PARENTAL ANIMALS)
Treatment-related changes were observed in the kidney and consisted of dilated pelvis (correlated with macroscopic examination), tubular dilation and/or basophilia, interstitial, intra-vascular, and/or tubular inflammatory cells sometimes associated with myxoid papilla stroma in some treated males and female given 5000 or 20000 ppm test item. The female given 20000 ppm had moderate suppurative nephritis associated with urothelium hyperplasia.

Dose descriptor:
NOEL
Effect level:
1 250 ppm (nominal)
Sex:
male/female
Basis for effect level:
other: (Corresponding to 79 or 103 mg/kg/day for males or females, respectively)
Clinical signs:
no effects observed
Mortality / viability:
no mortality observed
Body weight and weight changes:
effects observed, treatment-related
Sexual maturation:
not examined
Organ weight findings including organ / body weight ratios:
not examined
Gross pathological findings:
no effects observed
Histopathological findings:
not examined
VIABILITY (OFFSPRING)
It was considered that there was no effect of treatment with the test item on pup mortality

CLINICAL SIGNS (OFFSPRING)
No test item-related findings were noted.

BODY WEIGHT (OFFSPRING)
The body weight gain of male and female pups in the group treated at 20000 ppm was, respectively, 34% and 38% lower than controls on day 5 p.p.. This treatment-related effect coincided with the lower body weight gain recorded in parental animals in this group between days 1 and 5 p.p.. The terminal body weight differences were statistically significant for females only (p<0.05). No change on pups body weight was observed at the to lowest doses.

SEXUAL MATURATION (OFFSPRING)
not examined.

ORGAN WEIGHTS (OFFSPRING)
not examined.

GROSS PATHOLOGY (OFFSPRING)
not examined.

HISTOPATHOLOGY (OFFSPRING)
not examined.

OTHER FINDINGS (OFFSPRING)
Sex ratio: the percentage of male pups was between 44.2% and 54.4% on days 1 and 5 post-partum, which is considered to be within the normal range.
Reproductive effects observed:
not specified

Table 1: Body weight change (g) and percentage difference from controls (parental animals)

Sex Male Female
Concentration (ppm) 0 1250 5000 20000 0 1250 5000 20000
Treatment period (males) or premating period (females) days 1-15 +89 +76 (-15%) +80 (-10%) +51**(-43%) +34 +27 (-21%) +31 (-9%) +15# (-56%)
days 1-36 +165 +148 (-10%) +150 (-9%) +109# (-34%) na na na na
Gestation period GD 0-20 (overall mean) na na na na +165 +155 (-6%) +152 (-8%) +142 (-14%)
Lactation period LD 1-5 na na na na +22 +31 (+41%) +30 (+36%) +15 (-32%)

**: p<0.01, #: p<0.001, na: not applicable; GD: Gestation Day; LD: Lactation Day;

(): variations from controls.

Table 2: Mean achieved dosages (mg/kg/day)

Concentration (ppm)   1250 5000 (x4.0) 20000 (x4.0)
F0 generation Male premating (days 1-5) 79 319 (x4.0) 1099 (x4.0)
Female premating (days 1-5) 97 376 (x3.9) 1242 (x3.3)
gestation (GD 0-20) 90 356 (x4.0) 1285 (x3.6)
lactation (days 1-5 p.p.) 121 446 (x3.7) 1758 (x3.9)
Overall mean  103 393 1428

GD: Gestation Day, p.p.: post-partum,

(): ratio to the preceding concentration.

Conclusions:
Under the conditions of this test, 11-aminoundecanoic acid has no effect on mating, fertility, gestation or delivery.
Executive summary:

In a reproduction/developmental toxicity screening test following guideline OECD 421 (CIT, 2008), 11-aminoundecanoic acid (batch Nos. 0808204 and 0808295) was given to male and female Sprague-Dawley rats (10 animals per group) by dietary admixture for 2 weeks before mating, during mating, gestation and until day 4 post-partum, at the constant concentrations of 1250, 5000 or 20000 ppm. Another group of 10 males and 10 females received the untreated diet under the same experimental conditions and served as a control group. The concentrations of dietary admixture were checked in weeks 1, 4 and 6. Test item admixtures were within the acceptable concentration range of -7% to +19%. The nominal concentrations of 1250, 5000 or 20000 ppm corresponded to achieved dosages of 79, 319 or 1099 mg/kg/day for the males and 103, 393 or 1428 mg/kg/day for the females, thus demonstrating a satisfactory intake of the test item. There were no unscheduled deaths at any concentration. In-life findings recorded for the F0 animals were limited to one case of emaciated appearance in a male treated at 20000 ppm from day 36. Body weight was statistically significantly lower at 20000 ppm throughout the study and on day 5 of lactation in females treated at 5000 ppm. Food consumption was lower than controls during the study in goup treated at 20000 ppm and transiently in females treated at 5000 ppm (premating). No effects on mating, fertility, gestation or delivery were detected. There were no treatment-related effects on pup mortality or clinical signs. The body weight gain of pups at 20000 ppm was lower between days 1 and 5 of lactation. There were no relevant macroscopic abnormalities in pups sacrificed as scheduled or in pups found dead. There were no treatment-related effects on organ weights. Test item-related findings were observed in the kidneys of 2/10 males given 5000 ppm and 4/10 males and 1/10 females given 20000 ppm. These findings included dilated pelvis and granular kidneys with green content. Treatment-related changes were observed at microscopic examination in the kidney at 5000 and 20000 ppm and consisted of dilated pelvis, tubular dilatation and/or basophilia, interstitial, intra-vascular and/or tubular inflammatroy cells sometimes associated with myxoid papilla stroma. Based on the results of this study, the concentration of 1250 ppm was considered to be the No Observed Effect Level (NOEL) for parental toxicity and the concentration of 20000 ppm was considered to be the NOEL for reproductive performance.

Effect on fertility: via oral route
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
NOAEL
1 099 mg/kg bw/day
Study duration:
subchronic
Species:
rat
Effect on fertility: via inhalation route
Endpoint conclusion:
no study available
Effect on fertility: via dermal route
Endpoint conclusion:
no study available
Additional information

In a GLP reproduction/developmental toxicity screening test performed according to OECD 421 (CIT, 2008), the potential toxic effects of 11 -aminoundecanoic acid in rats were evaluated. This study provided information on all aspects of reproduction and development. The test substance was administered by dietary admixture for 15 days before mating, through mating, gestation and the beginning of the lactation period (until day 4 post-partum inclusive). The nominal concentrations used were 1250, 5000 or 20000 ppm corresponding to achieved dosages of 79, 319 or 1099 mg/kg/day for the males and 103, 393 or 1428 mg/kg/day for the females. No effects on mating, fertility, gestation or delivery were detected. There were no treatment-related effects on pup mortality or clinical signs and there were no relevant macroscopic abnormalities in pups sacrificed as scheduled or in pups found dead. No treatment-related effects were noted on testis or epididymis weights, at macroscopic examination or upon microscopic examination of the testes, epididymides or ovaries in treated rats. However, parental toxicity was observed substantiated by renal dose-related test item-related (dilated pelvis and granular kidneys) at 5000 and 20000 ppm test item associated with histopathological findings (tubular dilation and/or basophilia, interstitial, intra-vascular, and/or tubular inflammatory cells sometimes associated with myxoid papilla stroma in some treated males and female given 5000 or 20000 ppm test item). The female given 20000 ppm had moderate suppurative nephritis associated with urothelium hyperplasia. Based on these results, the concentration of 1250 ppm was considered to be the No Observed Effect Level (NOEL) for parental toxicity and the concentration of 20000 ppm was considered to be the NOEL for reproductive performance which is the highest dose tested.


Short description of key information:
Toxicity to reproduction: there is one key study performed according to OECD 421 where no effects were observed on mating, fertility, gestation or delivery.

Justification for selection of Effect on fertility via oral route:
No adverse effects were observed on the fertility performance.

Effects on developmental toxicity

Description of key information
Developmental toxicity:
. There is one developmental study performed in rats according to OECD 414 (Richard, 2001) with a NOAEL set at 172 mg/kg/day.
. There is a recent rabbit development study (Spézia, 2013). The NOAEL for maternal toxicity and fetal development was set at 100 mg/kg/day based on maternal toxicity associated with fetal findings at 300 mg/kg/day in group 4 while no fetal findings were observed in group 6 in the presence of maternal toxicity.
Link to relevant study records
Reference
Endpoint:
developmental toxicity
Type of information:
experimental study
Adequacy of study:
key study
Study period:
15 November 2011 - 6 December 2012
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Compliant to GLP and testing guidelines; adequate consistence between data, comments and conclusions.
Qualifier:
according to
Guideline:
OECD Guideline 414 (Prenatal Developmental Toxicity Study)
Deviations:
yes
Remarks:
the animals were acclimated to the study conditions for a period of 4 or 5 days before the beginning of the treatment period instead of a period of at least 5 days, 
GLP compliance:
yes (incl. certificate)
Limit test:
no
Species:
rabbit
Strain:
New Zealand White
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: breeder: Charles River Laboratories France (Châtillon-sur-Chalaronne, France)
- Age at study initiation: approximately 18 - 20 weeks old on the day of treatment
- Mean body weight at study initiation: 3421 g (range: 2795 g to 3805 g)
- Fasting period before study: no
- Housing: noryl cages
- Diet: SSNIFF R/M-H pelleted diet (free access)
- Water: tap water filtered with a 0.22 µm filter (free access)
- Acclimation period: the animals were acclimated to the study conditions for a period of 4 or 5 days before the beginning of the treatment period.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 2°C
- Humidity (%): 50 ± 20%
- Air changes (per hr): approximately 12 cycles/hour of filtered, non-recycled air
- Photoperiod (hrs dark / hrs light): 12 h/12 h (7:00 - 19:00).

IN-LIFE DATES: 14 November 2011 to 17 February 2012.
Route of administration:
oral: gavage
Vehicle:
CMC (carboxymethyl cellulose)
Details on exposure:
PREPARATION OF DOSING SOLUTIONS:
The test item was administered as a suspension in the vehicle. The test item was mixed with the required quantity of vehicle in order to achieve the concentrations of 10.0, 33.3 and 100.0 mg/mL. No correction factor was applied.
Homogeneity and stability of the dosage forms at 1 and 110 mg/mL were demonstrated for up to 10 days at +4°C and protected from light.
The test item dosage forms were prepared for use up to 10 days, stored at +4°C prior to use and delivered in brown flasks into crushed ice.

VEHICLE:
The vehicle was 0.5% carboxymethylcellulose aqueous solution (3 mL/kg) in drinking water prepared using:
- drinking water, treated by reverse osmosis using ELIX 5 plus apparatus (Millipore SA, Saint Quentin-en-Yvelines, France),
- carboxymethylcellulose, batch No. 100M0219V, supplied by Sigma (Saint-Quentin-Fallavier, France).

- Concentration in vehicle: 10.0, 33.3 and 100.0 mg/mL
- Amount of vehicle (if gavage): 3 mL/kg/day.
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
Type of method: HPLC-UV
Test item concentrations: The test item concentrations in the administered dosage forms analyzed in weeks 1 and 4 remained within an acceptable Range of variations (-8.5% to -0.4%) when compared to the nominal values (± 15%).
Dosage forms at 1 and 110 mg/mL were found to be homogeneous and stable after 10 days at +4°C and protected from light (CiToxLAB France Study No. 37498 AHS).
Details on mating procedure:
- Impregnation procedure: purchased time pregnant
- Proof of pregnancy: visual assessment (time-mated female rabbits obtained from the breeder).
Duration of treatment / exposure:
day 6 to day 28 post-coitum
Frequency of treatment:
Daily
Duration of test:
29 days
Remarks:
Doses / Concentrations:
0, 30, 100 and 300 mg/kg/day
Basis:
actual ingested
No. of animals per sex per dose:
22-23 females.
Control animals:
yes, concurrent vehicle
Details on study design:
- Dose selection rationale:
The dose-levels were selected in agreement with the Sponsor, following the results of a previous Maximum Tolerated Dose toxicity study by oral route (gavage) in non-pregnant rabbits and of a preliminary study of prenatal developmental toxicity by oral route (gavage) in rabbits (CIT/Study No. 37442 TSL).

In the Maximum Tolerated Dose toxicity study by oral route (gavage) in non-pregnant rabbits, animals (three female rabbits per group) received 300 or 1000 mg/kg/day for 5 days and after a 3-day treatment-free period, 300 or 600 mg/kg/day for 5 days, respectively. In group 1 (300 → 300 mg/kg/day), there were no clinical signs, no biologically significant effects on body weight or food consumption which might be attributed to the treatment with the test item. In group 2 (1000 → 600 mg/kg/day), 1/3 females had absence of feces and was emaciated on study termination (up to -915 g body weight loss (-26.4%) vs. body weight on study) and had no food consumption. Therefore, 1000 mg/kg/day was considered to be an excessive high dose-level. 600 mg/kg/day was selected as a high-dose for the preliminary study of prenatal developmental toxicity by oral route (gavage) in rabbits.

In the preliminary study of prenatal developmental toxicity by oral route (gavage) in rabbits, animals (six mated female rabbits per group) received 150, 300 or 600 mg/kg/day from day 6 to day 28 p.c.. There were no treatment-related clinical signs. In the high-dose group, 1/6 females with an history of prolonged body weight decrease and reduced food consumption, aborted on day 26 p.c. Slight to moderate dose related reduction in mean body weight were observed at 300 mg/kg/day (up to -7.3% vs. controls on day 24 p.c.) and at 600 mg/kg/day (up to -9.0% vs. controls on day 19 p.c.).
Reductions in mean food consumption were observed in all treated groups. These reductions were moderate at 150 and 300 mg/kg/day (up to -33.6% on days 12-15 p.c. and -25% on days 15-19 p.c. vs. controls, respectively) and severe at 600 mg/kg/day (-58.3% on days 6-9 p.c. vs. controls, p < 0.001 and -43.5% on days 9-12 p.c. vs. controls, p < 0.01). On hysterectomies and on external fetal examination, there were no treatment related findings.

Taking into account the abortion and the severe decrease in mean food consumption in the high dose-group, 600 mg/kg/day was considered to be an excessive high dose-level for a further definitive study.

Therefore, 300 mg/kg/day was selected as the high dose-level. The low-dose and mid dose-levels were selected using a ratio representing a 3-fold interval (i.e. 30 and 100 mg/kg/day).

- Rationale for animal assignment: stratified procedure base.

Study design in the main study:
Three groups of 22 mated female KBL New Zealand White rabbits were administered the test item, 11-aminundecanoic acid (batch 11.08.481), once daily from day 6 to day 28 p.c., by gavage at dosages of 30, 100 or 300 mg/kg/day (groups 2 to 4). An additional group of 22 mated females received the vehicle CMC, 0.5% under the same experimental conditions. A dosage volume of 3 ml/kg was used. Following poor clinical conditions during the acclimation period and following inconclusive results, it was decided at the request of the sponsor to repeat the experiment at the high dose-level (300 mg/kg/day) in order to ascertain whether or not the test item treatemnt was associated with fetal observations in recorded in group 4 (300 mg/kg/day) and wether or not the maternal condition prior to dosing had modify the potential susceptibility of the animals to the test item. Two additional groups of 23 and 22 mated female KB rabbits were administered under the same experimental conditions as above, etither the vehicle (group 5) or the test item at 300 mg/kg/day (group 6).
Maternal examinations:
CAGE SIDE OBSERVATIONS:
- Time schedule: at least twice a day during the treatment period.

DETAILED CLINICAL OBSERVATIONS:
- Time schedule: once a day during the treatment period.

BODY WEIGHT (GAIN):
- Time schedule: on days 2, 4, 5, 6, 9, 12, 15, 19, 24 and 29 p.c., and prior to premature sacrifice.

FOOD CONSUMPTION:
- Time schedule: on the following intervals: days 2-4, 4-5, 5-6, 6-9, 9-12, 12-15, 15-19, 19-24 and 24-29 p.c.

POST-MORTEM MACROSCOPIC EXAMINATION:
- Sacrifice on day 29 post-coitum.
- Examined: principal thoracic and abdominal organs.
Ovaries and uterine content:
The ovaries and uterine content were examined after termination, including::
- Gravid uterus weight
- Number of corpora lutea
- Number of implantations
- Number of early resorptions
- Number of late resorptions
- Number of uterine scars, evaluation of placenta.
Fetal examinations:
- External examinations: Yes: all per litter
- Soft tissue examinations: Yes: half per litter
- Skeletal examinations: Yes: half per litter
- Head examinations: Yes: half per litter
- Other : number dead and live, body weight, sex
Statistics:
Mean values were compared by a one-way analysis of variance and a Dunnett's test.
Percentage values were compared by a Fisher exact probability test.
Indices:
% Pre-implantation loss = 100 * (Number of corpora lutea - Number of implantation sites) / Number of corpora lutea
% Post-implantation loss = 100 * (Number of implantation sites - Number of live fetuses) / Number of implantation sites
Details on maternal toxic effects:
Maternal toxic effects:yes

Details on maternal toxic effects:
Mortality and Abortion

There were no mortality or abortion in the control and 100 mg/kg/day groups.

30 mg/kg/day
Dam No. X30044 was sacrificed on day 29 p.c. after aborting 15 fetuses (five placenta in the bedding). This female had an emaciated appearance from day 28 p.c.. At necropsy, there were no findings.
This finding was observed on day 29 p.c. as a results of an early delivery (total gestation period in rabbit is about 30 days). Therefore a treatment-related effect was considered unlikely.

300 mg/kg/day
Dam No. 30075 was sacrificed on day 13 p.c. after evidence of abortion (blood in the bedding). This female lostosed weight since day 2 p.c., had an absence of feces from day 8 p.c. and, an emaciated appearance on day 9 p.c. and from day 12 p.c.. This dam had no food consumption from the start of the study. At necropsy, there were no findings that could be related to treatment. Taking into account the sanitary status of this female during the pre treatment period, a relationship to the treatment was considered unlikely.
Dam No. 30083 was sacrificed on day 24 p.c. after evidence of abortion (blood in the bedding). This female had an emaciated appearance from day 23 p.c. and a cutaneous lesion on neck from day 20 p.c.. At necropsy, there were no findings that could be related to treatment. However, taking into account both the sanitary status of this female the days preceding abortion (significant decrease in body weight change during the period of days 6 to 15 p.c. or day 6 to 19 p.c., a relationship to the treatment cannot be ruled out.

When evaluating in detail individual data on body weight, body weight change or food consumption, there were the following observations:

- Dosing period:

Mean body weight changes on the period of days 6 to 9 p.c. in the 100 and 300 mg/kg/day groups were 26 and -2 g vs. 47 g in the control group, respectively. Mean food consumption in the 300 mg/kg/day group on the period of days 6 to 9 p.c. was 115 g vs. 148 g in the control group. These slight but not significant decreases in body weight and food consumption at 300 mg/kg/day associated with the abortion of one female are most probably indicative of maternal toxicity.

- Pre-dosing period:
On an individual basis, both in the control and test item-treated groups, there were dams with repeated episodes of weight loss and almost no food consumption (<10g/day) before treatment.
Episodes of body weight (BW) loss and almost no food consumption (FC<10 g/day) during days 2 to 6 before treatment before treatment as detailed below :


Females No. X30011:
BW: -305 g (-9.8%)
FC: 0 to 5 g/day (days 2 to 6 p.c.)

X30018:
BW: -50 g (-1.5%)
FC: 0 g/day (days 2 to 4 p.c.)
X30029:
BW: -325 g (-9.1%)
FC: 0 to 5 g/day (days 2 to 6 p.c.)

X30043:
BW: -450 g (-12.2%)
FC: 0 to 5 g/day (days 2 to 5 p.c.)

X30065:
BW: -325 g (-8.8%)
FC: 0 to 5 g/day (days 4 to 6 p.c.)

X30073:
BW: -65 g (-2.0%)
FC: 2 g/day (days 2 to 4 p.c.)

X30075:
BW: -410 g (-11.1%)
FC: 0 g/day (days 4 to 6 p.c.)

X30076:
BW: -225 g (-5.9%)
FC: 0 g/day (days 4 to 6 p.c.)

X30079:
BW: -245 g (-6.6%)
FC: 0 g/day (days 5 to 6 p.c.)





Dose descriptor:
NOAEL
Effect level:
100 mg/kg bw/day (actual dose received)
Based on:
test mat.
Basis for effect level:
other: maternal toxicity
Dose descriptor:
NOAEL
Effect level:
100 mg/kg bw/day (actual dose received)
Based on:
test mat.
Basis for effect level:
other: developmental toxicity
Details on embryotoxic / teratogenic effects:
Embryotoxic / teratogenic effects:yes

Details on embryotoxic / teratogenic effects:
When compared with the control group, there were no evidence of adverse effects of the treatment with the test item on reproductive parameters,
numbers of corpora lutea or implantations.

Fetal weight and sex :
There was no evidence of an adverse effect of the treatment with the test item on mean fetal body weights or fetal sex ratios.

External variation examination :
There were no effects on the incidence of external variations per fetuses or per litter.

External malformation examination:

Group 1 (vehicle control): one fetus (X30011-09) had a short tail (this finding was not observed in the Historical Control Data).

Group 3 (100 mg/kg/day): one fetus (X30057-03) had a spina bifida (occulta). This finding was not observed in control groups from other studies (Historical Control Data) and also observed in one fetus from the high-dose group but with no skeletal observation correlates. Spina bifida was also a common malformation reported in this animal species and strain (Charles River Historical Control Data, 2008-2010). Therefore a treatment-related effect was considered unlikely.

Group 4 (300 mg/kg/day): five fetuses from three different litters (X30076-01, X30081-07, X30087-01, X30087-03 and X30087-08) had a series of malformations (limb/extremities and/or central nervous system) for which a test item treatment-related effect cannot be excluded. All malformations were observed at litter and fetal incidences higher than the upper limit of the Historical Control Data.

Group 5 (vehicle control): fetuses had no external malformations.

Group 6 (300 mg/kg/day): fetuses had no external malformations.

Litter and fetal incidences of external malformations in group 4 were higher than the upper limit of the Historical Control Data while not external malformations were recorded at the same dose-level in group 6. Therefore, a relationship to the treatment was not excluded.


Soft tissues malformation
There were no soft tissue malformations in fetuses from group (vehicle control group), and, from group 2 (30 mg/kg/day).

Group 3 (100 mg/kg/day): one fetus (X30065-03) had a retroesophageal subclavian artery. This malformation was previously recorded in one fetus in control group 5 and in control groups from other studies at higher litter and fetal incidences (Historical Control Data). Therefore a relationship to treatment with the test item was considered unlikely.

Group 4 (300 mg/kg/day): one fetus (X30088-02) had an absent subclavian artery. This malformation was previously recorded in the Historical Control Data. Two fetuses from the same litter (X30087-08 and X30087-03) had a series of malformations (cardiovascular defects, misshapen cerebrum, open diaphragm, absent, gall bladder and/or adrenals). Some of these malformations were also recorded in the Historical Control Data (cardiovascular effects, absent gallbladder) or are regularly reported in public literature (Morita et al., 1987 and Stadler et al., 1983). However, for other malformations recorded in these fetuses (misshapen cerebrum, open diaphragm, absent adrenals) a test item treatment-related effect can not be excluded.

Group 6 (300 mg/kg/day): two fetuses from the same litter (X30116-02 and X30116-05) had malpositioned kidneys with short ureters. Malpositioned kidney is a malformation previously recorded in Historical Control Data.


Skeletal examination:
There were no singificant changes at cartilage examination and the incidence. The types of fetal variations findings recorded in groups 4 and 6 (300 mg/kg/day) are generally considered to represent slightly delayed ossification rather than dysmorphogenic changes.
There were no effect that could be related to the treatment at 30 and 100 mg/kg/day as the incidence of changes were either not dose-related or within the historical control data.

Group 2 (30 mg/kg/day): there were no skeletal malformations recorded at incidence higher than the upper limit of the Historical Control Data.

Group 3 (100 mg/kg/day): absent cervical hemivertebra(e) (fetus X30052-03), absent thoracic hemivertebra(e) (fetuses X30046-07, X30065-04, X30065-05) and absent rib(s) (fetus X30046 07) were not observed in the control group and not recorded in control groups from other studies (Historical Control Data). These findings were not observed in the higher dose group and only recorded in a few fetuses. Therefore, these findings were not considered to be treatment-related.


Group 4 (300 mg/kg/day): findings not recorded in the control group or in control groups from other studies (Historical Control Data) concerned two fetuses from two different litters: X30076 02 with absent thoracic hemivertebra(e) and X30087-03 with multiple severe malformations (cervical vertebra(e) with open arch, misshapen scapula, absent sternum and a series of abnormalities of limb and extremities). Based on litter incidences, a test item treatment-related effect cannot be excluded.

Group 6 (300 mg/kg/day): findings not recorded in the control groups (misaligned and fused lumbar vertebra(e) or extra sternebra) concerned the same fetuses. These malformations were recorded at incidences lower than the upper limit of the Historical Control Data, therefore a test item treatment-related effect was considered unlikely.

Abnormalities:
not specified
Developmental effects observed:
not specified
Conclusions:
The NOAELs obtained in the oral developmental toxicity study in rabbits are:
- maternal toxicity: 100 mg/kg/day
- development: 100 mg/kg/day
Executive summary:

The objective of this prenatal developmental toxicity study performed according to guideline OECD 414 was to evaluate the potential toxic effects of the test item, 11-AMINOUNDECANOIC ACID,on the pregnant female and on embryonic and fetal development following daily oral administration (gavage) to pregnant female rabbits from implantation to the day prior to the scheduled hysterectomy (day 6 to day 28 post‑coitum inclusive).

Three groups of 22 mated female KBL NewWhite rabbits were administered the test item, 11-AMINOUNDECANOIC ACID(batch No. 11.08.481), once daily from day 6 to day 28p.c., by gavage, at dosages of 30, 100 or 300 mg/kg/day (groups 2 to 4). An additional group of 22 mated females received the vehicle, 0.5% (w/v) carboxymethylcellulose, under the same experimental conditions and acted as the control group (group 1).A dose volume of 3 mL/kg/day was used.

In order to ascertain whether or not the test-item treatment was associated with fetal observations recorded in the high-dose group 4,it was decided at the request of the sponsor to complete the study by repeating administration at the high dose-level. Two additional groups of 23 and 22 mated female KBL New Zealand White rabbits were administered either the vehicle (group 5) or the test item, 11-AMINOUNDECANOIC ACID (batch No. 11.08.481) at 300 mg/kg/day (group 6) under the same experimental conditions as above.

Results

 

On an individual basis, both in control group 1 and test item-treated groups 2 to 4, there were dams with repeated episodes of weight loss and almost no food consumption (<10 g/day) before treatment. 

The inclusion of the two additional groups (group 5 and group 6) revealed/confirmed that the animals entering the study in groups 1 to 4 were not at the expected health conditions (body weight losses, reduced or absence of food consumption during the predosing period).

 

At termination on day 29p.c., there were 20, 21, 22, 17, 19 and 21 dams with live fetuses in groups 1 to 6, respectively.

 In the 30 mg/kg/day group 2, no findings were observed that were considered to be related to the treatment with the test item.

 In the 100 mg/kg/day group 3,the findings recorded in this dose group were observed at a low incidence and only in a few fetuses. Most findings were also recorded in the Historical Control Data at comparable incidences. Therefore it was concluded that the effects observed in this group were not related to treatment with the test item.

In the 300 mg/kg/day group 4, there were:

.         Maternal toxicity indicated by:

o      one aborted female on day 13p.c.and one aborted female on day 24p.c.,

o      not statistically significant decreases (especially recorded during the period of days 6 to 9 p.c.) on mean body weight, mean body weight change or on mean food consumption.

.         Fetal toxicity indicated by:

o      slightly higher incidences of external, soft tissue and skeletal malformations.

 

In the 300 mg/kg/day group 6, there were:

.         Maternal toxicity indicated by:

o      statistically significant reductions in mean body weight and mean body weight change,

o      statistically significant decreases in mean food consumption.

.         No fetal toxicity.

Conclusion

 

The test item, 11-AMINOUNDECANOIC ACID (batch No. 11.08.481), was administered by gavage, once daily, from days 6 to 28 p.c., inclusive, to mated female KBL New Zealand White rabbits at dosages of 30, 100 or 300 mg/kg/day.

Slight to moderate maternal toxicity was recorded in females given 300 mg/kg/day (group 4 and 6) while no maternal toxicity was recorded at 30 and 100 mg/kg/day.

The oral administration of 11-AMINOUNDECANOIC ACID to the dams at all dose-levels had no influence on gestational parameters. Conception rate, mean number of corpora lutea, total implantations, resorptions and live fetuses, fetal sex ratio and the pre-and the post-implantation losses were all unaffected by the treatment.

Fetal examinations revealed no influence of the test compound on sex ratio and fetal body weight. The fetal findings associated with slight maternal toxicity observed in group 4 (300 mg/kg/day) were not recorded in group 6 (300 mg/kg/day) in the presence of maternal toxicity, therefore a treatment related effect could not be ascertained.

 

On the basis of the results obtained in this study:

.         The No Observed Adverse Effect Level (NOAEL) for maternal parameters was considered to be 100 mg/kg/day, based on decreased mean body weight gain and food consumption at 300 mg/kg/day,

.         The NOAEL for embryo-fetal development was considered to be 100 mg/kg/day based on dubious findings recorded at 300 mg/kg/day in a context of maternal toxicity.

 

Effect on developmental toxicity: via oral route
Endpoint conclusion:
adverse effect observed
Dose descriptor:
NOAEL
100 mg/kg bw/day
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

1- Prenatal developmental toxicity study (CIT, 2001): the potential toxic effects of 11 -aminoundecanoic acid on embryonic and fetal development following daily oral administration (dietary admixture) to pregnant female rats during the entire period of gestation: from fertilization to approximately one day before the expected day of parturition (day 2 to day 19 post-coitum inclusive) were evaluated. 11 -aminoundecanoic acid was administered at the constant concentrations of 2000, 6000 and 18000 ppm (equivalent to a daily intake of 172, 520 and 1394 mg/kg/day, respectively). As a consequence of poor appetite, possibly due to the unpalatability of the dietary admixture at 18000, lower food consumption and body weight gain were recorded in dams at this high concentration. A slight retardation of growth/skeletal development substantiated by a decrease in the incidence of two skeletal variations (unossified 4 th metacarpal and unossified 6 th sternebrae) was observed in offsprings at 6000 ppm and particularly 18000 ppm (dose-level at which a slight reduction in fetal body weight was also noted), which was considered as an indirect effect of maternal toxicity. Under the conditions of this test, the No Observed Effect Level for embryofetal development was established at 2000 ppm (i.e 172 mg/kg/day); 6000 ppm (520 mg/kg/day) can be considered as a No Observed Adverse Effect Level.

2- Prenatal rabbit developmental study (Spézia, 2013),

The objective of this prenatal developmental toxicity study performed according to guideline OECD 414 was to evaluate the potential toxic effects of the test item,11-AMINOUNDECANOIC ACID,on the pregnant female and on embryonic and fetal development following daily oral administration (gavage) to pregnant female rabbits from implantation to the day prior to the scheduled hysterectomy (day 6 to day 28post‑coituminclusive) (Spézia, 2013).

Three groups of 22 mated female KBL NewWhite rabbits were administered the test item, 11-AMINOUNDECANOIC ACID(batch No. 11.08.481), once daily from day 6 to day 28p.c., by gavage, at dosages of 30, 100 or 300 mg/kg/day (groups 2 to 4). An additional group of 22 mated females received the vehicle, 0.5% (w/v) carboxymethylcellulose, under the same experimental conditions and acted as the control group (group 1).A dose volume of 3 mL/kg/day was used.

As the health conditions of the animals before start of dosing were not optimal among the groups and in order to ascertain whether or not the test-item treatment was associated with fetal observations recorded in the high-dose group 4 and , it was decided to complete the study by repeating administration at the high dose-level. Two additional groups of 23 and 22 mated female KBL New Zealand White rabbits were administered either the vehicle (group 5) or the test item, 11-AMINOUNDECANOIC ACID (batch No. 11.08.481) at 300 mg/kg/day (group 6) under the same experimental conditions as above.

Slight to moderate maternal toxicity was recorded in females given 300 mg/kg/day (group 4 and 6) while no maternal toxicity was recorded at 30 and 100 mg/kg/day.

The oral administration of 11-AMINOUNDECANOIC ACID to the dams at all dose-levels had no influence on gestational parameters. Conception rate, mean number of corpora lutea, total implantations, resorptions and live fetuses, fetal sex ratio and the pre-and the post-implantation losses were all unaffected by the treatment.

Fetal examinations revealed no influence of the test compound on sex ratio and fetal body weight.

The inclusion of the two additional groups (group 5 and group 6) revealed/confirmed that the animals entering the study in groups 1 to 4 were not at the expected health conditions (body weight losses, reduced or absence of food consumption during the predosing period).

The fetal findings associated with slight maternal toxicity observed in group 4 (300 mg/kg/day) were NOT recorded in group 6 (300 mg/kg/day) in the presence of maternal toxicity, therefore a treatment related effect could not be ascertained.

On the basis of the results obtained in this study:

.         the No Observed Adverse Effect Level (NOAEL) for maternal parameters was considered to be 100 mg/kg/day, based on decreased mean body weight gain and food consumption at 300 mg/kg/day,

.         the NOAEL for embryo-fetal development was considered to be 100 mg/kg/day based on dubious findings recorded at 300 mg/kg/day in a context of maternal toxicity.


Justification for selection of Effect on developmental toxicity: via oral route:
In the rabbit developmental study, slightly higher incidence of fetal findings associated with slight maternal toxicity was observed at the high dose-level. The fetal findings were not observed in the complementary group at the limit dose-level of 300 mg/kg/day while maternal toxicity was recorded.
In the rat developmental study, the No effect Level for embryofetal development was established at 2000 ppm (i.e. 172 mg/kg/day) and 6000 ppm of 11-AMINOUNDECANOIC ACID in diet (equivalent to 520 mg/kg/day) was established as a No Observed Adverse Effect Level for both maternal and embryofetal toxicity in rats based on significant decrease in fetal body weight and food consumption and delayed ossification at the limit dose of 18000 ppm (1394 mg/kg/day).

Justification for classification or non-classification

Based on the lack of effect on male and female fertility, reproduction organs and prenatal parameters at dose-level of 20000 ppm (1099 mg/kg/day for males and 1428 mg/kg/day for females), no classification for fertility is warranted. In the rat developmental toxicity study the NOAEL for maternal toxicity and developmental toxicity was 520 mg/kg/day. Minor fetal findings (delayed ossification) were observed at the maximal highest tested dose-level of 1394 mg/kg/day together with maternal toxicity. In the rabbit developmental toxicity study, the NOAEL for maternal toxicity and developmental toxicity was 100 mg/kg/day based on decreased maternal body weight and minor fetal effects observed at 300 mg/kg/day. The minor developmental effects observed were associated with maternal toxicity at the MTD dose-level in the rat (1394 mg/kg/day) and at the highest dose tested of 300 mg/kg/day in the rabbits. Furthermore, no fetal findings attributed to the test item were observed in the rabbit study in the complementary group given the high dose-level (300 mg/kg/day) although there was maternal toxicity. No fetal or maternal effects that could be attirbuted to the test item were observed at the low and mid dose-levels in both species. Because the fetal findings were minor and associated with maternal toxicity at the MTD dose-levels (1394 mg/kg/day) in the rats and at the limit dose of 300 mg/kg/day in the rabbits, no classification is warranted for developmental toxicity.