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EC number: 266-587-2 | CAS number: 67151-63-7
Developmental toxicity / teratogenicity
Administrative data
- Endpoint:
- developmental toxicity
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Study period:
- 07 November 2017 - 15 March 2018
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- guideline study
Data source
Reference
- Reference Type:
- study report
- Title:
- Unnamed
- Year:
- 2 018
- Report date:
- 2018
Materials and methods
Test guidelineopen allclose all
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 414 (Prenatal Developmental Toxicity Study)
- Version / remarks:
- Adopted 22 January 2001
- Deviations:
- no
- Qualifier:
- according to guideline
- Guideline:
- EPA OPPTS 870.3700 (Prenatal Developmental Toxicity Study)
- Version / remarks:
- August 1998
- Deviations:
- no
- Qualifier:
- according to guideline
- Guideline:
- other: Japanese Ministry of Agriculture, Forestry and Fisheries Testing guideline for Toxicology studies, 12 NouSan No 8147
- Version / remarks:
- 24 November 2000
- Deviations:
- no
- GLP compliance:
- yes
- Limit test:
- no
Test material
- Reference substance name:
- 1-[bis[3-(dimethylamino)propyl]amino]propan-2-ol
- EC Number:
- 266-587-2
- EC Name:
- 1-[bis[3-(dimethylamino)propyl]amino]propan-2-ol
- Cas Number:
- 67151-63-7
- Molecular formula:
- C13H31N3O
- IUPAC Name:
- 1-Bis(3-(dimethylamino)propylamino)-2-propanol
- Test material form:
- liquid
Constituent 1
- Specific details on test material used for the study:
- SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: PFW160721
- Expiration date of the lot/batch: 30 December 2018
- Purity : ca. 98.32%
STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: Stored at ambient temperature/humidity in darkness; may be used/formulated in light
- Solubility and stability of the test substance in the solvent/vehicle: For the purpose of the study the test item was prepared at the appropriate concentrations as a solution in Distilled Water. The stability and homogeneity of the test item formulations were determined by Envigo Research Limited, Shardlow, UK Analytical Services. The formulations were stable for at least 21 days.
OTHER SPECIFICS: No correction for purity was made.
Test animals
- Species:
- rat
- Strain:
- Sprague-Dawley
- Remarks:
- Crl:CD (SD) IGS BR
- Details on test animals or test system and environmental conditions:
- TEST ANIMALS
- Source: Charles River (UK) Limited, Margate, Kent
- Age at study initiation: no data
- Weight at study initiation: At the start of treatment the females weighed 204 to 299g.
- Fasting period before study: no data
- Housing: The animals were housed individually in solid-floor polypropylene cages with stainless steel mesh lids furnished with softwood flakes (Datesand Ltd., Cheshire, UK).
- Diet (e.g. ad libitum): ad libitum; A pelleted diet (Rodent 2018C Teklad Global Certified Diet, Envigo RMS (UK) Limited, Oxon, UK) was used.
- Water (e.g. ad libitum): ad libitum, Mains drinking water was supplied from polycarbonate bottles attached to the cage.
- Acclimation period: no data
ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 3°C
- Humidity (%): 50 ± 20%
- Air changes (per hr): at least 15 air changes per hour
- Photoperiod (hrs dark / hrs light): twelve hours continuous light and twelve hours darkness
IN-LIFE DATES: From: 01 December 2017 (first day of treatment) To: 20 December 2017 (final day of necropsy)
Administration / exposure
- Route of administration:
- oral: gavage
- Vehicle:
- water
- Remarks:
- distilled water
- Details on exposure:
- PREPARATION OF DOSING SOLUTIONS:
Formulations are stable for at least 21 days and were therefore prepared twice and stored at approximately 4 °C in the dark.
VEHICLE
- Concentration in vehicle: 0, 2.5, 10 and 25 mg/mL
- Treatment volume 10 mL/kg - Analytical verification of doses or concentrations:
- yes
- Details on analytical verification of doses or concentrations:
- The test item concentration in the test samples was determined by gas chromatography (GC) using an external standard technique. The test item gave a chromatographic profile consisting of a single peak.
The analytical procedure was successfully validated with respect to specificity of chromatographic analysis, linearity of detector response, method accuracy and precision.
The homogeneity and stability was confirmed for test item in distilled water formulations at nominal concentrations of 2.5 mg/mL and 25 mg/mL when stored refrigerated for 21 days.
The mean concentrations of test item in test formulations analyzed for the study were within ± 10% of nominal concentrations, confirming accurate formulation.
The results indicate that the prepared formulations were within 3% of the nominal concentration. - Details on mating procedure:
- Animals were delivered in two batches containing females prior to Day 3 of gestation. The day that positive evidence of mating was observed was designated Day 0 of gestation.
- Duration of treatment / exposure:
- between Days 3 and 19 of gestation
- Frequency of treatment:
- daily
- Duration of test:
- From Day 3 to Day 20 of gestation.
Doses / concentrationsopen allclose all
- Dose / conc.:
- 25 mg/kg bw/day (nominal)
- Remarks:
- low treatment group
- Dose / conc.:
- 100 mg/kg bw/day (nominal)
- Remarks:
- Intermediate treatment group
- Dose / conc.:
- 250 mg/kg bw/day (nominal)
- Remarks:
- high treatment group
- No. of animals per sex per dose:
- 24
- Control animals:
- yes, concurrent vehicle
- Details on study design:
- - Dose selection rationale: The dose levels were chosen in collaboration with the Sponsor and were based on previous toxicity data (Envigo Research Limited, Study Number XX96MF). The preliminary oral (gavage) pre-natal development toxicity study in the rat was performed as dose range finder for the OECD 414 study. Following dose levels were recommended for use in the main prenatal developmental toxicity study: 0 (Control), 25, 100 and 250 mg/kg bw/day. Based on the results of the study, dose levels of 0 (Control), 25, 100 and 250 mg/kg bw/day were recommended for use in the planned prenatal developmental toxicity study.
The highest dose level should produce an effect on body weight gain and the low and intermediate dose levels follow the recommendations of the OECD 414 guideline for a 2 to 4-fold interval between dose levels.
- Rationale for animal assignment: The animals were randomly allocated to treatment groups using a randomization procedure based on stratified body weight to ensure similarity between the treatment groups.
Examinations
- Maternal examinations:
- DETAILED CLINICAL OBSERVATIONS: Yes
Following arrival, all animals were examined for overt signs of toxicity, ill-health or behavioral changes once daily during the gestation period. Additionally, during the dosing period, observations were recorded immediately before and soon after dosing and one hour post dosing. All observations were recorded.
BODY WEIGHT: Yes
Individual body weights were recorded on Day 3 (before the start of treatment) and on Days 5, 6, 7, 8, 11, 14 and 17 of gestation. Body weights were also recorded for animals at terminal kill (Day 20).
FOOD CONSUMPTION: Yes
Food consumption was recorded for each individual animal at Day 3, 5, 8, 11, 14, 17 and 20 of gestation.
WATER CONSUMPTION: Yes
Water intake was observed daily by visual inspection of the water bottles for any overt changes.
POST-MORTEM EXAMINATIONS: Yes
All animals were killed by carbon dioxide asphyxiation followed by cervical dislocation on Day 20 of gestation. All animals were subjected to a full external and internal examination and any macroscopic abnormalities were recorded.
- Ovaries and uterine content:
- The ovaries and uteri of pregnant females were removed, examined and the following data recorded:
i) Number of corpora lutea
ii) Number, position and type of intrauterine implantation
iii) Fetal sex
iv) External fetal appearance
v) Fetal weight
vi) Placental weight
vii) Gravid uterus weight
The uteri of any apparently non-pregnant females were immersed in 0.5% ammonium polysulphide solution to reveal evidence of implantation.
Implantation types were divided into:
Early Death: No visible distinction between placental/decidual tissue and embryonic tissue
Late Death: Separate embryonic/fetal and placental tissue visible
Dead Fetus: A fetus that had died shortly before necropsy. These were included as late deaths for reporting purposes
All implantations and viable fetuses were numbered according to their intrauterine position as follows (as an example):
Left Horn Cervix Right Horn
L1 L2 L3 L4 L5 L6 L7 L8 R1 R2 R3 R4 R5 R6 R7 R8
V1 V2 V3 V4 V5 V6 V7 V8 V9 V10 V11 V12 V13 V14 V15 V16
V = viable fetus - Fetal examinations:
- The fetuses were killed by subcutaneous injection of a suitable barbiturate agent. Fetuses from each litter were divided into two groups and examined for skeletal alterations and soft tissue alterations. Alternate fetuses were identified using an indelible marker and placed in Bouin’s fixative. Fetuses were subsequently transferred to distilled water and examined for visceral anomalies under a low power binocular microscope and then stored in 10% Buffered Formalin. The remaining fetuses were identified using cardboard tags marked with chinagraph pencil and placed into 70% IMS in distilled water. The fetuses were subsequently eviscerated, processed and the skeletons stained with alizarin red S before being transferred to 50% glycerol for examination of skeletal development and anomalies and storage.
- Statistics:
- The following parameters were analyzed statistically, where appropriate, using the test methods outlined below:
Female body weight change, food consumption and gravid uterus weight: Shapiro Wilk normality test and Bartlett’s test for homogeneity of variance and one way analysis of variance, followed by Dunnett’s multiple comparison test or, if unequal variances were observed, on alternative multiple comparison test.
All caesarean necropsy parameters and fetal parameters: Kruskal-Wallis non-parametric analysis of variance; and a subsequent pairwise analysis of control values against treated values using the Mann-Whitney ‘U’ test, where significance was seen.
Fetal evaluation parameters, including skeletal or visceral findings: Kruskal-Wallis non-parametric analysis of variance and Mann-Whitney ‘U’ test.
Probability values (p) are presented as follows:
p<0.001 ***
p<0.01 **
p<0.05 *
p≥0.05 (not significant) - Indices:
- Percentage pre-implantation loss was calculated as:
[(number of corpora lutea-number of implantations)/number of corpora lutea] x 100
Percentage post-implantation loss was calculated as:
[(number of implantations - number of live fetuses)/number of implantations] x 100
Sex ratio:
Sex ratio was calculated as:
% male fetuses (sex ratio) = (number of male fetuses/total number of fetuses) x 100
Results and discussion
Results: maternal animals
General toxicity (maternal animals)
- Clinical signs:
- effects observed, non-treatment-related
- Description (incidence and severity):
- There were no significant adverse clinical signs evident in treated females.
One female treated with 250 mg/kg bw/day showed increased salivation post dosing on Day 5 only. A further female from this treatment group had pilo-erection on Day 8 only. In isolation, these findings were considered to be incidental and of no toxicological importance. - Dermal irritation (if dermal study):
- not examined
- Mortality:
- no mortality observed
- Description (incidence):
- There were no unscheduled deaths.
- Body weight and weight changes:
- effects observed, treatment-related
- Description (incidence and severity):
- Females treated with 250 mg/kg bw/day showed a statistically significant reduction (p<0.01-0.001) in body weight gain between Days 4 and 11 of gestation. Body weight for these females was also statistically significantly reduced (p<0.05-0.01) from Day 8 onwards. Cumulative body weight gain was statistically significantly reduced (p<0.05-0.001) in these females throughout the treatment period and body weight and body weight gain when adjusted for gravid uterus weight was also statistically significantly reduced (p<0.05 and p<0.001 respectively) when compared to controls.
A statistically significant reduction (p<0.05) in body weight gain was evident in females treated with 100 mg/kg bw/day between Days 8 and 11 of gestation. A statistically significant reduction (p<0.05) in cumulative body weight gain was also evident in these females between Days 3 and 11 and Days 3 and 17. Body weight gain for these females during the remaining periods was comparable to controls.
Body weight gain during gestation, including after adjustment for the contribution of the gravid uterus, was considered to be unaffected by treatment at 25 mg/kg bw/day. See section 'Additional information on results' for more details. Historical control data for the parameters is described, when available. - Food consumption and compound intake (if feeding study):
- effects observed, treatment-related
- Description (incidence and severity):
- Females treated with 250 mg/kg bw/day showed a statistically significant reduction (p<0.01-0.001) in food consumption throughout the treatment period.
No differences as compared to the control group were detected for food consumption in females treated with 100 or 25 mg/kg bw/day. - Food efficiency:
- not examined
- Water consumption and compound intake (if drinking water study):
- no effects observed
- Description (incidence and severity):
- No differences as compared to the control group were detected for water consumption.
- 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:
- effects observed, non-treatment-related
- Description (incidence and severity):
- No treatment related macroscopic abnormalities were detected in females treated with 250, 100 or 25 mg/kg bw/day.
One female treated with 100 mg/kg bw/day had a scab on the right shoulder and one control female had an enlarged right kidney. These findings were considered to be incidental findings. - Neuropathological findings:
- not examined
- Histopathological findings: non-neoplastic:
- not examined
- Histopathological findings: neoplastic:
- not examined
- Other effects:
- not specified
- Details on results:
- At 100 mg/kg bw/day, treatment was associated with slightly lower body weight gains between Days 8 and 11 of gestation and lower cumulative body weight gain between Days 3 and 11 and Days 3 and 17 of gestation. Body weight gain for these females during the remaining periods was comparable to controls. No macroscopic necropsy findings were apparent for adult females at this dosage. Given the transient nature of the effects on body weight gain, this dosage is considered to represent a No Observed Adverse Effect Level (NOAEL) for the pregnant female.
Maternal developmental toxicity
- Number of abortions:
- no effects observed
- Description (incidence and severity):
- There was no treatment-related effect observed in the pre- and post-implantation loss in all dose groups, when comparing to the control group:- preimplantation loss: 13.3%, 13.6%, 10.5%, 13.2% at 0, 25, 100 and 250 mg/kg bw/day-post-implantation loss: 0.5%, 0.9%, 1.4%, 1.2% at 0, 25, 100 and 250 mg/kg bw/day.All data was within the historical control range available for this species and strain. See section 'Additional information on results' for more details
- Pre- and post-implantation loss:
- no effects observed
- Description (incidence and severity):
- There was no treatment-related effect observed in the pre- and post-implantation loss in all dose groups, when comparing to the control group:
- preimplantation loss: 13.3%, 13.6%, 10.5%, 13.2% at 0, 25, 100 and 250 mg/kg bw/day
- post-implantation loss: 0.5%, 0.9%, 1.4%, 1.2% at 0, 25, 100 and 250 mg/kg bw/day.
All data was within the historical control range available for this species and strain. See section 'Additional information on results' for more details. - Total litter losses by resorption:
- no effects observed
- Description (incidence and severity):
- There were no total litter losses by resorption observed. See Section 'Additional information on results' for more details.
- Early or late resorptions:
- no effects observed
- Description (incidence and severity):
- There were no treatment-related effects on early or late resorptions observed. See Section 'Additional information on results' for more details.
- Dead fetuses:
- no effects observed
- Description (incidence and severity):
- There were no treatment-related effects on the number of dead fetuses observed. See Section 'Additional information on results' for more details.
- Changes in pregnancy duration:
- no effects observed
- Description (incidence and severity):
- There was no treatment-related, toxicologically relevant effect on the number of pregnant dams per dose group. The number of pregnant females per dose group were 24/24, 23/24, 23/24 and 22/24 at 0, 25, 100 and 250 mg/kg bw/day, resp. No historical control data is available for this parameter. See section 'Additional information on results' for more details
- Changes in number of pregnant:
- no effects observed
- Description (incidence and severity):
- There was no treatment-related, toxicologically relevant effect on the number of pregnant dams per dose group. The number of pregnant females per dose group were 24/24, 23/24, 23/24 and 22/24 at 0, 25, 100 and 250 mg/kg bw/day, resp. No historical control data is available for this parameter. See section 'Additional information on results' for more details
- Other effects:
- not specified
- Details on maternal toxic effects:
- There was no obvious effect of maternal treatment on litter data as assessed by numbers of implantations, in-utero offspring survival (as assessed by the mean numbers of early or late resorptions), live litter size, sex ratio or pre- or post-implantation losses at 25, 100 or 250 mg/kg bw/day.
Intergroup differences for mean fetal, litter or placental weights did not indicate any obvious effects of maternal treatment at 25, 100 or 250 mg/kg bw/day.
Statistical analysis of the data did not reveal any significant intergroup differences.
Effect levels (maternal animals)
- Key result
- Dose descriptor:
- NOAEL
- Effect level:
- 100 mg/kg bw/day (nominal)
- Based on:
- test mat.
- Basis for effect level:
- body weight and weight gain
Maternal abnormalities
- Abnormalities:
- no effects observed
Results (fetuses)
- Fetal body weight changes:
- no effects observed
- Description (incidence and severity):
- There is no treatment-related effect on the mean fetal body weight, mean male fetal body weight or mean female fetal body weight. See section 'Additional information on results' for further details. Available historical control data is described.
- Reduction in number of live offspring:
- no effects observed
- Description (incidence and severity):
- There is no treatment-related effect on the number of live offspring. The percentage of live offspring was calculated as the number of live implants - the number of implants x 100. This means for the dose groups: 99.3%, 99.3%, 98.6% and 98.5% for the control, 25, 100 and 250 mg/kg bw/day. See section 'Additional information on results' for further details. The available historical control data is described.
- Changes in sex ratio:
- no effects observed
- Description (incidence and severity):
- There was no treatment-related effect observed on sex ratio. See Section 'Additional information on results' for more details. Historical control data is described, when available.
- Changes in litter size and weights:
- no effects observed
- Description (incidence and severity):
- There was no treatment-related effects observed on litter weight or litter size. See Section 'Additional information on results' for more details. Historical control data is described, when available.
- Changes in postnatal survival:
- no effects observed
- Description (incidence and severity):
- There was no treatment-related effects observed on postnatal survival. See Section 'Additional information on results' for more details. Historical control data is described, when available.
- External malformations:
- no effects observed
- Description (incidence and severity):
- No treatment-related effect on external malformations was observed in all dose groups. See section 'additional information on results' for further details. Available historical control data is described.
- Skeletal malformations:
- effects observed, non-treatment-related
- Description (incidence and severity):
- A statistically significant increase (p<0.05-0.01) in the number of fetuses/litters showing incomplete ossification of the parietal, interparietal, hyoid, sacral arch, sternebra and femur were evident at 250 mg/kg bw/day. The effect on the sternebra also extended to fetuses/litters at 100 and 25 mg/kg bw/day. A statistically significant increase (p<0.05) in the number of fetuses/litters showing incomplete ossification of the zygomatic process of squamosal were evident at 25 and 250 mg/kg bw/day. A statistically significant increase (p<0.05) in the number of fetuses/litters showing incomplete ossification of the hyoid was evident at 100 mg/kg bw/day. With the exception of the sacral arch and sternebra, true dose related responses were not evident in the remaining parameters and group mean values were generally within historical control ranges (excluding interparietal, sacral arch and femur). In the absence of any particular pattern of abnormal skeletal development of skeletal structures affecting treated fetuses and in the absence of an effect on mean fetal weight, the observation of isolated affected skeletal structures can be considered unlikely to represent true developmental abnormalities.
See Section 'Additional information on results' for further details. Available historical control data is described. - Visceral malformations:
- no effects observed
- Description (incidence and severity):
- No treatment-related effect on visceral malformations was observed in all dose groups. See section 'additional information on results' for further details. Available historical control data is described.
- Other effects:
- not specified
- Details on embryotoxic / teratogenic effects:
- No treatment-related changes were detected in the offspring parameters measured or on embryofetal development. The ‘No Observed Adverse Effect Level’ (NOAEL) for developmental toxicity was therefore considered to be 250 mg/kg bw/day.
Effect levels (fetuses)
- Key result
- Dose descriptor:
- NOAEL
- Effect level:
- 250 mg/kg bw/day (nominal)
- Based on:
- test mat.
- Sex:
- male/female
- Basis for effect level:
- other: no treatment-related changes up to 250 mg/kg bw/day
Fetal abnormalities
- Abnormalities:
- no effects observed
Overall developmental toxicity
- Developmental effects observed:
- no
Any other information on results incl. tables
Table 1: Summary of Female Performance: number of pregnant and non-pregnant dams per dose group
Category |
Number of Females at Dose Level (mg/kg bw/day) |
|||
0 (Control) |
25 |
100 |
250 |
|
Initial Group Size |
24 |
24 |
24 |
24 |
Non-Pregnant |
0 |
1 |
1 |
2 |
Pregnant |
24 |
23 |
23 |
22 |
° No historical data available
Table 2: Group Mean Litter Data Values: pre- and postimplantation loss, number and percent
Dose Level (mg/kg bw/day) |
|
Number of Corpora Lutea |
Number of Implants |
Number of Embryonic/Fetal Deaths |
Implantation Loss |
Number of live implants | |||||
Early |
Late |
Total |
Pre |
Post |
Male | Female | Total | ||||
0 (Control) |
mean |
16.3 |
14.0 |
0.0 |
0.0 |
0.1 |
13.3 |
0.5 |
6.0 | 7.9 | 13.9 |
sd# |
2.1 |
1.6 |
0.2 |
0.2 |
0.3 |
8.2 |
1.8 |
1.8 | 2.0 | 1.5 | |
number of animals |
24 |
24 |
24 |
24 |
24 |
24 |
24 |
24 | 24 | 24 | |
25 |
mean |
16.7 |
14.4 |
0.1 |
0.0 |
0.1 |
13.6 |
0.9 |
6.7 | 7.6 | 14.3 |
sd# |
1.8 |
1.8 |
0.3 |
0.0 |
0.3 |
7.9 |
2.5 |
1.7 | 1.9 | 1.9 | |
number of animals |
23 |
23 |
23 |
23 |
23 |
23 |
23 |
23 | 23 | 23 | |
100 |
mean |
15.9 |
14.2 |
0.2 |
0.0 |
0.2 |
10.5 |
1.4 |
7.3 | 6.7 | 14.0 |
sd# |
1.5 |
1.5 |
0.5 |
0.2 |
0.6 |
7.6 |
3.9 |
1.9 | 1.7 | 1.5 | |
number of animals |
23 |
23 |
23 |
23 |
23 |
23 |
23 |
23 | 23 | 23 | |
250 |
mean |
15.9 |
13.7 |
0.1 |
0.1 |
0.2 |
13.2 |
1.2 |
6 | 7.5 | 13.5 |
sd# |
2.1 |
2.0 |
0.3 |
0.3 |
0.5 |
8.9 |
3.2 |
2.6 | 2.2 | 1.9 | |
number of animals |
22 |
22 |
22 |
22 |
22 |
22 |
22 |
22 | 22 | 22 | |
| Historical data | range$ | 11 - 20 | 9 - 18 | 0 - 2 | 0 - 4 | 0 - 4 | 0 - 31.8 | 0 - 6.9 | 3 - 10 | 3 - 10 | 10 - 17 |
| mean | 15 | 14 | 0 | 0 | 0 | 11.4 | 1.3 | 6 | 7 | 13 | |
| sd# | 2 | 2 | 0 | 1 | 1 | 10.2 | 2.8 | 2 | 2 | 2 | |
| number of animals | 194 | 200 | 210 | 210 | 210 | 203 | 198 | 201 | 196 | 203 | |
# sd: range = minimum to maximum
$ range = mean + / - 2 standard deviations
pre-implantation loss = number of corpora lutea - number of implants; pre-implantation loss % = pre-implantation loss / number of corpora lutea x 100; post-implantation loss = number of implants - number of live implants (total); postimplantation loss % = post implantation loss / number of implants x 100
Table 3: Group Mean Body Weight Values:
Dose Level (mg/kg bw/day) |
|
Body Weight (g) on Day of Gestation |
|||||||
3 |
4 |
5 |
8 |
11 |
14 |
17 |
20 |
||
0 (Control) |
mean |
252.8 |
257.2 |
263.0 |
276.1 |
292.8 |
311.9 |
339.1 |
382.5 |
sd |
23.1 |
24.4 |
25.8 |
26.7 |
28.8 |
29.2 |
31.8 |
33.6 |
|
number of animals |
24 |
24 |
24 |
24 |
24 |
24 |
24 |
24 |
|
25 |
mean |
251.8 |
258.4 |
263.7 |
276.7 |
293.3 |
312.8 |
338.0 |
383.3 |
sd |
16.4 |
19.4 |
19.0 |
18.7 |
20.4 |
21.0 |
21.2 |
24.4 |
|
number of animals |
23 |
23 |
23 |
23 |
23 |
23 |
23 |
23 |
|
100 |
mean |
248.6 |
253.4 |
257.7 |
269.1 |
283.1 |
302.4 |
326.8 |
370.7 |
sd |
13.2 |
15.3 |
15.3 |
15.5 |
17.0 |
16.7 |
17.9 |
21.0 |
|
number of animals |
23 |
23 |
23 |
23 |
23 |
23 |
23 |
23 |
|
250 |
mean |
252.0 |
253.6 |
254.3 |
260.1* |
268.7** |
290.5** |
316.1** |
358.9* |
sd |
18.6 |
18.2 |
17.5 |
18.6 |
18.9 |
19.1 |
20.6 |
26.5 |
|
number of animals |
22 |
22 |
22 |
22 |
22 |
22 |
22 |
22 |
|
| Historical data | range | 204 - 291 | / ° | 214 - 306 | 225 - 321 | 243 - 341 | 258 - 361 | 283 - 396 | 322 - 447 |
| mean | 247 | / ° | 260 | 273 | 292 | 310 | 340 | 385 | |
| sd | 22 | / ° | 23 | 24 | 25 | 26 | 28 | 31 | |
| number of animals | 288 | / ° | 290 | 290 | 289 | 290 | 289 | 290 | |
° For Day 4 no historical data is available
° *p<0.05; ** p<0.01
Table 4: Group Mean Body Weight Change Values
Dose Level (mg/kg bw/day) |
|
Body Weight Change (g) during Days of Gestation |
||||||
3 to 4 |
4 to 5 |
5 to 8 |
8 to 11 |
11 to 14 |
14 to 17 |
17 to 20 |
||
0 (Control) |
mean |
4.4 |
5.8 |
13.1 |
16.8 |
19.1 |
27.2 |
43.4 |
sd |
4.1 |
3.7 |
3.4 |
4.1 |
3.5 |
4.9 |
5.9 |
|
number of animals |
24 |
24 |
24 |
24 |
24 |
24 |
24 |
|
25 |
mean |
6.6 |
5.3 |
13.1 |
16.5 |
19.6 |
25.2 |
45.3 |
sd |
3.7 |
3.0 |
4.4 |
3.4 |
3.8 |
3.5 |
6.8 |
|
number of animals |
23 |
23 |
23 |
23 |
23 |
23 |
23 |
|
100 |
mean |
4.9 |
4.3 |
11.3 |
14.0* |
19.3 |
24.4 |
43.9 |
sd |
3.9 |
2.9 |
5.0 |
5.6 |
4.7 |
4.8 |
5.8 |
|
number of animals |
23 |
23 |
23 |
23 |
23 |
23 |
23 |
|
250 |
mean |
1.6 |
0.6*** |
5.8*** |
8.6** |
21.9 |
25.6 |
42.7 |
sd |
3.9 |
4.9 |
5.2 |
16.4 |
17.3 |
6.5 |
10.6 |
|
number of animals |
22 |
22 |
22 |
22 |
22 |
22 |
22 |
|
° * p<0.05; ** p<0.01; *** p<0.001
Table 4a: Group Mean Body Weight Change Values - Historical Data
Days 3 -5 |
Days 5 - 6 |
Days 6 - 7 |
Days 7 - 8 |
Days 8 - 11 |
Days 11 - 14 |
Days 14 - 18 |
Days 17 - 20 |
||
Historical data: body weight change (g) |
range |
3 - 22 |
-5 - 10 |
-2 - 13 |
-2 - 11 |
10 - 29 |
10 - 27 |
20 - 40 |
32 - 53 |
mean |
12 |
3 |
5 |
5 |
19 |
18 |
30 |
44 |
|
sd |
5 |
4 |
4 |
3 |
5 |
4 |
5 |
6 |
|
number of animals |
286 |
128 |
205 |
298 |
293 |
288 |
288 |
289 |
Table 5: Group Mean Gravid Uterus Weight and Adjusted Body Weight and Body Weight Change Values
Dose Level (mg/kg bw/day) |
|
Body Weight (g) on Days of Gestation |
Body Weight Change (g) during Days of Gestation |
Gravid Uterus Weight |
Adjusted |
Adjusted |
|
3 |
20 |
3-20 |
|||||
0 (Control) |
mean |
252.8 |
382.5 |
129.7 |
82.880 |
299.6 |
46.8 |
sd |
23.1 |
33.6 |
12.9 |
8.641 |
29.4 |
10.1 |
|
number of animals |
24 |
24 |
24 |
24 |
24 |
24 |
|
25 |
mean |
251.8 |
383.3 |
131.4 |
85.789 |
297.5 |
45.6 |
sd |
16.4 |
24.4 |
10.7 |
9.994 |
22.1 |
10.7 |
|
number of animals |
23 |
23 |
23 |
23 |
23 |
23 |
|
100 |
mean |
248.6 |
370.7 |
122.1 |
82.770 |
287.9 |
39.4 |
sd |
13.2 |
21.0 |
14.0 |
7.550 |
17.4 |
10.7 |
|
number of animals |
23 |
23 |
23 |
23 |
23 |
23 |
|
250 |
mean |
252.0 |
358.9 |
106.9 |
78.508 |
280.4* |
28.4*** |
sd |
18.6 |
26.5 |
12.1 |
8.685 |
24.4 |
12.4 |
|
number of animals |
22 |
22 |
22 |
22 |
22 |
22 |
|
°* p<0.05; ** p<0.01; *** p<0.001
° Available Historical Data:
- Gravid Uterus Weight (g):
~ Range: 63.56 - 106.59 ~ Mean: 85.08 +/- 10.76 ~ Number of animals: 286
- Adjusted Body Weight Day 20 (g):
~ Range: 247 - 353 ~ Mean 300 +/- 26 ~ Number of animals: 286
- Adjusted Body Weight Change Days 5 - 20 (g):
~ Range: 20 - 64 ~ Mean 45 +/- 11 ~ Number of animals: 126
Table 6: Mean Number and Percent of Live Offspring
| Dose Level (mg/kg bw/day) | Number of Live Implants | % Male Fetuses | total live implants / number of implants x 100 | |||
| Male | Female | Total | ||||
| 0 (Control) | mean | 6.0 | 7.9 | 13.9 | 43.1 | 99.3 |
| sd | 1.8 | 2.0 | 1.5 | 12.7 | / | |
| 25 | mean | 6.7 | 7.6 | 14.3 | 46.8 | 99.3 |
| sd | 1.7 | 1.9 | 1.9 | 11.2 | / | |
| 100 | mean | 7.3 | 6.7 | 14.0 | 51.9 | 98.6 |
| sd | 1.9 | 1.7 | 1.5 | 11.7 | / | |
| 250 | mean | 6.0 | 7.5 | 13.5 | 43.9 | 98.5 |
| sd | 2.6 | 2.2 | 1.9 | 17.1 | / | |
| Historical data | mean | 6 | 7 | 13 | 48.5 | / |
| sd | 2 | 2 | 2 | 31.1 | / | |
Table 7: Mean Foetal / Pup Body Weight by Sex and Sexes Combined
| Dose Level (mg/kg bw/day | Mean Male Fetal Weight (g) | Mean Female Fetal Weight (g) | Mean Fetal Weight (g) | Mean Placental Weight (g) | Litter Weight (g) | Total Placental Weight (g) | |
| 0 (Control) | mean | 3.959 | 3.747 | 3.840 | 0.541 | 53.382 | 7.537 |
| sd | 0.201 | 0.152 | 0.168 | 0.046 | 5.739 | 1.081 | |
| number of animals | 24 | 24 | 24 | 24 | 24 | 24 | |
| 25 | mean | 3.923 | 3.733 | 3.825 | 0.548 | 54.584 | 7.803 |
| sd | 0.314 | 0.270 | 0.287 | 0.053 | 7.193 | 1.117 | |
| number of animals | 23 | 23 | 23 | 23 | 23 | 23 | |
| 100 | mean | 3.922 | 3.686 | 3.805 | 0.566 | 52.922 | 7.883 |
| sd | 0.282 | 0.313 | 0.290 | 0.071 | 5.116 | 1.234 | |
| number of animals | 23 | 23 | 23 | 23 | 23 | 23 | |
| 250 | mean | 3.778 | 3.618 | 3.691 | 0.560 | 49.832 | 7.475 |
| sd | 0.345 | 0.320 | 0.319 | 0.106 | 6.747 | 1.038 | |
| number of animals | 22 | 22 | 22 | 22 | 22 | 22 | |
| Historical data | range | 3.64 - 4.59 | 3.36 - 4.45 | 3.46 - 4.54 | 0.4 - 0.73 | 35.43 - 71.14 | 5.24 - 9.71 |
| mean | 4.11 | 3.9 | 4 | 0.57 | 53.28 | 7.47 | |
| sd | 0.24 | 0.27 | 0.27 | 0.08 | 8.93 | 1.12 | |
| number of animals | 200 | 203 | 203 | 203 | 202 | 202 |
° range = mean + / - 2 standard deviations
Table 8: Summary Incidence of Fetal External Findings
External Findings |
Dose level (mg/kg bw/day) |
|||||||||||
0 (Control) |
25 |
100 |
250 |
|||||||||
Number of fetuses (litters) examined |
||||||||||||
334 (24) |
329 (23) |
321 (23) |
298 (22) |
|||||||||
NF |
NL |
%† |
NF |
NL |
%† |
NF |
NL |
%† |
NF |
NL |
%† |
|
Small fetus |
5 |
4 |
1.5 |
6 |
5 |
1.8 |
7 |
5 |
2.0 |
13 |
4 |
5.6 |
Small placenta |
3 |
2 |
0.9 |
4 |
3 |
1.2 |
1 |
1 |
0.3 |
1 |
1 |
0.3 |
Pale fetus |
1 |
1 |
0.3 |
0 |
0 |
0.0 |
3 |
3 |
0.9 |
1 |
1 |
0.3 |
Large placenta |
0 |
0 |
0.0 |
1 |
1 |
0.3 |
5 |
2 |
1.5 |
12 |
2 |
5.4 |
Pale placenta |
0 |
0 |
0.0 |
0 |
0 |
0.0 |
0 |
0 |
0.0 |
1 |
1 |
0.3 |
Total |
8 |
5 |
2.4 |
8 |
6 |
2.3 |
14 |
8 |
4.1 |
15 |
5 |
6.4 |
%†: Group mean percent
NF: Number of Fetuses
NL: Number of litters
Table 9: Summary Incidence of Fetal Visceral Findings
Visceral Findings |
Dose Level (mg/kg bw/day) |
|||||||||||
0 (Control) |
25 |
100 |
250 |
|||||||||
Number of Fetuses (litters) Examined |
||||||||||||
172 (24) |
169 (23) |
166 (23) |
156 (22) |
|||||||||
NF |
NL |
%† |
NF |
NL |
%† |
NF |
NL |
%† |
NF |
NL |
%† |
|
External |
|
|
|
|
|
|
|
|
|
|
|
|
Hemorrhage |
1 |
1 |
0.6 |
1 |
1 |
0.5 |
0 |
0 |
0.0 |
0 |
0 |
0.0 |
Situs inversus |
1 |
1 |
0.6 |
0 |
0 |
0.0 |
0 |
0 |
0.0 |
0 |
0 |
0.0 |
Head |
|
|
|
|
|
|
|
|
|
|
|
|
Tongue - short |
0 |
0 |
0.0 |
0 |
0 |
0.0 |
1 |
1 |
0.7 |
0 |
0 |
0.0 |
Rugae - non-uniform patterning |
5 |
5 |
2.8 |
11 |
8 |
6.6 |
7 |
7 |
4.3 |
13 |
9 |
7.9 |
Brain - olfactory ventricle - enlarged |
0 |
0 |
0.0 |
0 |
0 |
0.0 |
1 |
1 |
0.5 |
0 |
0 |
0.0 |
Brain - cerebral aqueduct - enlarged |
0 |
0 |
0.0 |
0 |
0 |
0.0 |
2 |
1 |
1.1 |
0 |
0 |
0.0 |
Abdomen |
|
|
|
|
|
|
|
|
|
|
|
|
Liver - additional lobe between right and left median |
2 |
2 |
1.2 |
1 |
1 |
0.7 |
0 |
0 |
0.0 |
0 |
0 |
0.0 |
Liver - papillary process - reduced in size |
1 |
1 |
0.6 |
0 |
0 |
0.0 |
0 |
0 |
0.0 |
0 |
0 |
0.0 |
Umbilical artery - left-sided |
1 |
1 |
0.5 |
0 |
0 |
0.0 |
0 |
0 |
0.0 |
0 |
0 |
0.0 |
Testis - partially undescended |
1 |
1 |
0.5 |
1 |
1 |
0.6 |
0 |
0 |
0.0 |
1 |
1 |
0.6 |
Ureter - kinked |
28 |
13 |
16.3 |
17 |
11 |
10.2 |
10 |
7 |
5.7 |
12 |
8 |
7.3 |
Ureter - dilated |
23 |
11 |
13.4 |
12 |
8 |
7.4 |
7 |
4 |
4.0* |
3 |
3 |
1.9** |
Renal pelvic cavitation - increased |
9 |
5 |
5.1 |
10 |
7 |
5.9 |
4 |
4 |
2.3 |
11 |
6 |
6.4 |
Renal papilla - absent |
2 |
2 |
1.2 |
1 |
1 |
0.6 |
1 |
1 |
0.5 |
4 |
2 |
2.5 |
Renal medulla - reduced in size |
0 |
0 |
0.0 |
0 |
0 |
0.0 |
1 |
1 |
0.5 |
0 |
0 |
0.0 |
Renal medulla - absent |
0 |
0 |
0.0 |
0 |
0 |
0.0 |
0 |
0 |
0.0 |
1 |
1 |
0.6 |
Thorax |
|
|
|
|
|
|
|
|
|
|
|
|
Thyroid - Isthmus thickened |
0 |
0 |
0.0 |
0 |
0 |
0.0 |
2 |
1 |
1.1 |
0 |
0 |
0.0 |
Thyroid - parathyroid - enlarged |
0 |
0 |
0.0 |
0 |
0 |
0.0 |
1 |
1 |
0.5 |
0 |
0 |
0.0 |
Thymus - lobe partially undescended |
9 |
5 |
4.9 |
10 |
6 |
6.0 |
4 |
4 |
2.4 |
13 |
9 |
8.0 |
Lungs - right lobe - single lobe present |
1 |
1 |
0.6 |
0 |
0 |
0.0 |
0 |
0 |
0.0 |
0 |
0 |
0.0 |
%†: Group mean percent
NF: Number of Fetuses
NL: Number of litters
* p<0.05; ** p<0.01
Table 9a: Summary Incidence of Fetal Visceral Findings Continued
Visceral Findings |
Dose Level (mg/kg bw/day) |
|||||||||||
0 (Control) |
25 |
100 |
250 |
|||||||||
Number of Fetuses (litters) Examined |
||||||||||||
172 (24) |
169 (23) |
166 (23) |
156 (22) |
|||||||||
NF |
NL |
%† |
NF |
NL |
%† |
NF |
NL |
%† |
NF |
NL |
%† |
|
Thorax (continued) |
|
|
|
|
|
|
|
|
|
|
|
|
Atrium - enlarged |
1 |
1 |
0.6 |
0 |
0 |
0.0 |
1 |
1 |
0.6 |
0 |
0 |
0.0 |
Single atrioventricular valve |
1 |
1 |
0.6 |
0 |
0 |
0.0 |
0 |
0 |
0.0 |
0 |
0 |
0.0 |
Atrial septal defect |
1 |
1 |
0.6 |
0 |
0 |
0.0 |
0 |
0 |
0.0 |
0 |
0 |
0.0 |
Ventricle - enlarged |
1 |
1 |
0.6 |
0 |
0 |
0.0 |
0 |
0 |
0.0 |
0 |
0 |
0.0 |
Ventricular septal defect |
1 |
1 |
0.6 |
0 |
0 |
0.0 |
0 |
0 |
0.0 |
0 |
0 |
0.0 |
Total |
45 |
17 |
25.7 |
37 |
18 |
22.2 |
25 |
16 |
14.8 |
39 |
15 |
23.7 |
%†: Group mean percent
NF: Number of Fetuses
NL: Number of litters
Table 10: Summary Incidence of Fetal Skeletal Findings
Skeletal Findings |
Dose Level (mg/kg bw/day) |
|||||||||||
0 (Control) |
25 |
100 |
250 |
|||||||||
Number of Fetuses (litters) Examined |
||||||||||||
162 (24) |
160 (23) |
155 (23) |
142 (22) |
|||||||||
NF |
NL |
%† |
NF |
NL |
%† |
NF |
NL |
%† |
NF |
NL |
%† |
|
Skull |
|
|
|
|
|
|
|
|
|
|
|
|
Fontanelle (anterior) - large |
2 |
1 |
1.0 |
0 |
0 |
0.0 |
4 |
4 |
2.5 |
1 |
1 |
0.9 |
Nasal - incomplete ossification |
15 |
8 |
8.9 |
23 |
13 |
14.3 |
14 |
8 |
8.7 |
32 |
14 |
21.1 |
Nasal/Frontal - sutural bone |
1 |
1 |
0.6 |
1 |
1 |
0.5 |
0 |
0 |
0.0 |
0 |
0 |
0.0 |
Frontal - incomplete ossification |
3 |
3 |
1.7 |
4 |
4 |
2.5 |
4 |
4 |
2.4 |
12 |
9 |
7.8 |
Frontal - unossified area |
3 |
3 |
1.8 |
1 |
1 |
0.7 |
5 |
4 |
3.1 |
14 |
7 |
9.9 |
Frontal - fissure |
0 |
0 |
0.0 |
1 |
1 |
0.7 |
0 |
0 |
0.0 |
0 |
0 |
0.0 |
Parietal - incomplete ossification |
9 |
6 |
5.3 |
14 |
10 |
8.6 |
12 |
7 |
7.4 |
27 |
12 |
17.9* |
Parietal/Interparietal - sutural bone |
0 |
0 |
0.0 |
0 |
0 |
0.0 |
0 |
0 |
0.0 |
2 |
2 |
1.3 |
Interparietal - incomplete ossification |
32 |
13 |
19.9 |
27 |
12 |
16.9 |
26 |
13 |
16.6 |
54 |
17 |
37.2* |
Occipital (Supra-occipital) - incomplete ossification |
26 |
12 |
16.1 |
20 |
12 |
12.6 |
22 |
13 |
14.2 |
46 |
16 |
30.7 |
Occipital (Supra-occipital) - unossified area(s) |
11 |
9 |
7.1 |
10 |
8 |
6.3 |
9 |
6 |
5.7 |
6 |
6 |
4.0 |
Squamosal - incomplete ossification |
18 |
9 |
11.3 |
22 |
11 |
13.6 |
18 |
9 |
11.5 |
33 |
13 |
22.1 |
Squamosal - unossified area(s) |
0 |
0 |
0.0 |
1 |
1 |
0.5 |
2 |
2 |
1.2 |
4 |
3 |
2.5 |
Jugal - incomplete ossification |
5 |
4 |
2.9 |
11 |
8 |
7.3 |
6 |
6 |
4.2 |
17 |
9 |
11.7 |
Zygomatic process of maxilla - incomplete ossification |
9 |
7 |
5.5 |
19 |
11 |
12.4 |
18 |
10 |
11.7 |
25 |
11 |
17.4 |
Zygomatic process of maxilla -elongated |
1 |
1 |
0.6 |
0 |
0 |
0.0 |
0 |
0 |
0.0 |
0 |
0 |
0.0 |
Zygomatic process of squamosal - incomplete ossification |
0 |
0 |
0.0 |
5 |
4 |
3.0* |
1 |
1 |
0.6 |
9 |
5 |
6.2* |
Zygomatic process of squamosal - fused to jugal |
1 |
1 |
0.6 |
0 |
0 |
0.0 |
0 |
0 |
0.0 |
0 |
0 |
0.0 |
Premaxilla - incomplete ossification |
4 |
3 |
2.3 |
1 |
1 |
0.6 |
4 |
4 |
2.3 |
9 |
6 |
6.2 |
Hyoid - incomplete ossification |
10 |
9 |
5.8 |
16 |
11 |
9.7 |
24 |
15 |
15.7* |
17 |
8 |
11.1* |
Hyoid - not ossified |
21 |
12 |
12.5 |
17 |
12 |
9.7 |
26 |
13 |
16.6 |
30 |
14 |
19.2 |
Tympanic annulus - incomplete ossification |
0 |
0 |
0.0 |
0 |
0 |
0.0 |
1 |
1 |
0.6 |
0 |
0 |
0.0 |
%†: Group mean percent
NF: Number of Fetuses
NL: Number of litters
* p<0.05
Table 10a: Summary Incidence of Fetal Skeletal Findings - Continued
Skeletal Findings |
Dose Level (mg/kg bw/day) |
|||||||||||
0 (Control) |
25 |
100 |
250 |
|||||||||
Number of Fetuses (litters) Examined |
||||||||||||
162 (24) |
160 (23) |
155 (23) |
142 (22) |
|||||||||
NF |
NL |
%† |
NF |
NL |
%† |
NF |
NL |
%† |
NF |
NL |
%† |
|
Skull (continued) |
|
|
|
|
|
|
|
|
|
|
|
|
Presphenoid - incomplete ossification |
1 |
1 |
0.7 |
5 |
2 |
2.6 |
2 |
2 |
1.2 |
3 |
2 |
1.9 |
Presphenoid - not ossified |
0 |
0 |
0.0 |
2 |
2 |
1.1 |
0 |
0 |
0.0 |
3 |
3 |
1.9 |
Vertebral column |
|
|
|
|
|
|
|
|
|
|
|
|
Odontoid - ossification present |
3 |
3 |
1.8 |
0 |
0 |
0.0 |
0 |
0 |
0.0 |
0 |
0 |
0.0 |
Ventral arch of vertebra 1 - ossification present |
30 |
16 |
18.6 |
47 |
22 |
29.8 |
40 |
19 |
26.8 |
49 |
16 |
34.8 |
Cervical (neural) arch - incomplete ossification |
13 |
7 |
7.5 |
10 |
7 |
6.7 |
9 |
8 |
5.7 |
8 |
6 |
5.4 |
Cervical (neural) arch - fused |
0 |
0 |
0.0 |
0 |
0 |
0.0 |
0 |
0 |
0.0 |
1 |
1 |
0.6 |
Thoracic centrum - incomplete ossification |
5 |
4 |
2.9 |
5 |
4 |
3.4 |
2 |
2 |
1.2 |
5 |
4 |
3.5 |
Thoracic centrum - not ossified |
1 |
1 |
0.6 |
0 |
0 |
0.0 |
2 |
2 |
1.2 |
0 |
0 |
0.0 |
Thoracic centrum - bipartite ossification |
2 |
2 |
1.1 |
3 |
3 |
2.1 |
1 |
1 |
0.6 |
2 |
2 |
1.7 |
Thoracic centrum - dumb-bell-shaped |
18 |
12 |
11.4 |
15 |
11 |
9.2 |
12 |
9 |
8.1 |
20 |
12 |
14.0 |
Thoracic centrum - asymmetrically ossified |
4 |
2 |
2.3 |
2 |
2 |
1.6 |
4 |
4 |
2.7 |
2 |
2 |
1.2 |
Lumbar centrum - incomplete ossification |
1 |
1 |
0.6 |
1 |
1 |
0.5 |
0 |
0 |
0.0 |
0 |
0 |
0.0 |
Lumbar (neural) arch - incomplete ossification |
0 |
0 |
0.0 |
0 |
0 |
0.0 |
0 |
0 |
0.0 |
1 |
1 |
0.6 |
Sacral centrum - incomplete ossification |
0 |
0 |
0.0 |
0 |
0 |
0.0 |
1 |
1 |
0.5 |
0 |
0 |
0.0 |
Sacral (neural) arch - incomplete ossification |
25 |
12 |
15.0 |
28 |
12 |
17.7 |
33 |
16 |
21.2 |
51 |
17 |
34.9* |
Sacral (neural) arch - not ossified |
0 |
0 |
0.0 |
0 |
0 |
0.0 |
1 |
1 |
0.5 |
1 |
1 |
0.8 |
Caudal vertebrae - less than 4 ossified |
39 |
18 |
23.6 |
39 |
15 |
23.9 |
43 |
15 |
27.4 |
59 |
16 |
39.6 |
Number of pre-sacral vertebrae = 25/27 |
2 |
2 |
1.3 |
0 |
0 |
0.0 |
1 |
1 |
0.6 |
0 |
0 |
0.0 |
Ribs |
|
|
|
|
|
|
|
|
|
|
|
|
Ossification centre - associated with 7th cervical vertebra |
0 |
0 |
0.0 |
0 |
0 |
0.0 |
1 |
1 |
0.6 |
0 |
0 |
0.0 |
14th rib - extra - associated with 1st lumbar vertebra |
0 |
0 |
0.0 |
0 |
0 |
0.0 |
1 |
1 |
0.6 |
0 |
0 |
0.0 |
Ossification centre - associated with 1st lumbar vertebra |
5 |
5 |
3.2 |
9 |
3 |
5.8 |
6 |
5 |
4.0 |
2 |
2 |
1.3 |
%†: Group mean percent
NF: Number of Fetuses
NL: Number of litters
* p<0.05
Table 10b: Summary Incidence of Fetal Skeletal Findings - Continued
Skeletal Findings |
Dose Level (mg/kg bw/day) |
|||||||||||
0 (Control) |
25 |
100 |
250 |
|||||||||
Number of Fetuses (litters) Examined |
||||||||||||
162 (24) |
160 (23) |
155 (23) |
142 (22) |
|||||||||
NF |
NL |
%† |
NF |
NL |
%† |
NF |
NL |
%† |
NF |
NL |
%† |
|
Ribs (continued) |
|
|
|
|
|
|
|
|
|
|
|
|
One or more ribs - wavy |
1 |
1 |
0.7 |
1 |
1 |
0.6 |
0 |
0 |
0.0 |
1 |
1 |
0.6 |
One or more ribs - thickened |
1 |
1 |
0.7 |
0 |
0 |
0.0 |
1 |
1 |
0.6 |
0 |
0 |
0.0 |
Rib - short |
1 |
1 |
0.7 |
1 |
1 |
0.5 |
0 |
0 |
0.0 |
0 |
0 |
0.0 |
Rib - rudimentary |
1 |
1 |
0.6 |
0 |
0 |
0.0 |
0 |
0 |
0.0 |
0 |
0 |
0.0 |
Rib - fused |
1 |
1 |
0.6 |
0 |
0 |
0.0 |
0 |
0 |
0.0 |
0 |
0 |
0.0 |
Rib - no articulation point |
1 |
1 |
0.6 |
1 |
1 |
0.5 |
0 |
0 |
0.0 |
0 |
0 |
0.0 |
Costal cartilage - misaligned |
4 |
4 |
2.4 |
2 |
2 |
1.0 |
2 |
2 |
1.2 |
1 |
1 |
0.6 |
Costal cartilage - not fused to sternebra |
28 |
12 |
17.9 |
16 |
8 |
10.3 |
14 |
9 |
9.2 |
13 |
10 |
9.6 |
Sternebrae |
|
|
|
|
|
|
|
|
|
|
|
|
Sternebra - incomplete ossification |
0 |
0 |
0.0 |
5 |
4 |
3.0* |
5 |
5 |
3.1* |
8 |
7 |
5.4** |
Sternebra - not ossified |
2 |
2 |
1.2 |
0 |
0 |
0.0 |
2 |
2 |
1.2 |
0 |
0 |
0.0 |
Sternebra - bipartite ossification |
1 |
1 |
0.6 |
0 |
0 |
0.0 |
1 |
1 |
0.9 |
0 |
0 |
0.0 |
Sternebra - misaligned |
2 |
2 |
1.2 |
3 |
3 |
1.7 |
6 |
6 |
4.0 |
5 |
4 |
3.6 |
Sternebra - misshapen |
1 |
1 |
0.6 |
0 |
0 |
0.0 |
0 |
0 |
0.0 |
0 |
0 |
0.0 |
Xiphoid cartilage - partially split |
4 |
4 |
2.5 |
6 |
6 |
3.8 |
11 |
7 |
8.0 |
6 |
4 |
4.4 |
Pectoral Girdle |
|
|
|
|
|
|
|
|
|
|
|
|
Scapula - bent |
0 |
0 |
0.0 |
0 |
0 |
0.0 |
1 |
1 |
0.5 |
0 |
0 |
0.0 |
Scapula - misshapen |
3 |
3 |
1.8 |
0 |
0 |
0.0 |
1 |
1 |
0.7 |
4 |
4 |
2.7 |
Pelvic Girdle |
|
|
|
|
|
|
|
|
|
|
|
|
Ischium - incomplete ossification |
1 |
1 |
0.6 |
2 |
2 |
1.3 |
1 |
1 |
0.6 |
6 |
5 |
4.0 |
Pubis - not ossified |
1 |
1 |
0.6 |
2 |
2 |
1.1 |
1 |
1 |
0.6 |
3 |
3 |
2.2 |
Pubis - incomplete ossification |
8 |
6 |
4.6 |
5 |
5 |
3.1 |
12 |
7 |
7.6 |
15 |
11 |
9.9 |
Table 10c: Summary Incidence of Fetal Skeletal Findings - Continued
Skeletal Findings |
Dose Level (mg/kg bw/day) |
|||||||||||
0 (Control) |
25 |
100 |
250 |
|||||||||
Number of Fetuses (litters) Examined |
||||||||||||
162 (24) |
160 (23) |
155 (23) |
142 (22) |
|||||||||
NF |
NL |
%† |
NF |
NL |
%† |
NF |
NL |
%† |
NF |
NL |
%† |
|
Forelimbs |
|
|
|
|
|
|
|
|
|
|
|
|
Metacarpal - not ossified |
56 |
19 |
34.3 |
46 |
16 |
28.4 |
63 |
19 |
39.5 |
83 |
18 |
56.4 |
Metacarpal - incomplete ossification |
2 |
2 |
1.2 |
2 |
2 |
1.2 |
3 |
3 |
1.9 |
9 |
6 |
5.8 |
Forepaw phalanges - 1 or more - ossified |
8 |
4 |
4.8 |
14 |
7 |
8.5 |
10 |
6 |
6.3 |
8 |
6 |
5.6 |
Humerus - incomplete ossification |
1 |
1 |
0.6 |
3 |
3 |
1.8 |
2 |
2 |
1.2 |
7 |
5 |
4.8 |
Humerus - hole |
3 |
3 |
1.8 |
0 |
0 |
0.0 |
1 |
1 |
0.6 |
0 |
0 |
0.0 |
Humerus - bent |
0 |
0 |
0.0 |
0 |
0 |
0.0 |
1 |
1 |
0.5 |
0 |
0 |
0.0 |
Hindlimbs |
|
|
|
|
|
|
|
|
|
|
|
|
Metatarsal - 1st - ossified |
1 |
1 |
0.8 |
1 |
1 |
0.5 |
0 |
0 |
0.0 |
0 |
0 |
0.0 |
Metatarsal - not ossified |
0 |
0 |
0.0 |
0 |
0 |
0.0 |
2 |
2 |
1.2 |
0 |
0 |
0.0 |
Metatarsal - incomplete ossification |
1 |
1 |
0.6 |
0 |
0 |
0.0 |
2 |
2 |
1.2 |
2 |
2 |
1.6 |
Femur - incomplete ossification |
11 |
7 |
6.9 |
6 |
4 |
3.8 |
17 |
8 |
10.1 |
30 |
14 |
20.6** |
Total |
136 |
24 |
84.3 |
132 |
23 |
82.3 |
134 |
23 |
86.8 |
135 |
22 |
95.2 |
%†: Group mean percent
NF: Number of Fetuses
NL: Number of litters
* p<0.05; ** p<0.01
Table 11 Caesarea necropsy findings
| number of females | V | ED | LD | DF | NP | |
| control | 24 | 22 | 1 | 1 | ||
| 25 mg/kg bw/d | 24 | 20 | 3 | 1 | ||
| 100 mg/kg bw/d | 24 | 20 | 3 | 1 | 1 | |
| 250 mg/kg bw/d | 24 | 19 | 2 | 1 | 1 | 2 |
V: viable fetuses; ED: early death (no visible distinction between placental/decidual tissue and embryonic tissue); LD: late death: seperate embryonic/fetal and placental tissue visible; DF: dead fetus: a fetus that died shortly before necropsy; NP: non pregnant
Applicant's summary and conclusion
- Conclusions:
- The oral (gavage) administration of the test substance to pregnant rats during gestation at dose levels of 25, 100 and 250 mg/kg bw/day resulted in reductions in body weight gain and food consumption throughout the treatment period in females treated with 250 mg/kg bw/day. Although a slight reduction in body weight gain and cumulative body weight gain was noted for the 100 mg/kg bw/day dose group between Days 8 and 11 and Days 3 and 11 and 3 and 17 respectively, the effects were minimal and body weight gains for the remainder of the periods were comparable to controls. Consequently, 100 mg/kg bw/day was considered to represent the No Observed Adverse Effect Level (NOAEL) for the pregnant female.
No treatment-related changes were detected in the offspring parameters measured or on embryofetal development. The ‘No Observed Adverse Effect Level’ (NOAEL) for developmental toxicity was therefore considered to be 250 mg/kg bw/day.
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