Registration Dossier

Toxicological information

Developmental toxicity / teratogenicity

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Administrative data

Endpoint:
developmental toxicity
Type of information:
experimental study
Adequacy of study:
key study
Study period:
28 June 2016 - 21 July 2016
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2016
Report Date:
2016

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 414 (Prenatal Developmental Toxicity Study)
Version / remarks:
January 2001
Deviations:
no
Qualifier:
according to
Guideline:
EU Method B.31 (Prenatal Developmental Toxicity Study)
Version / remarks:
May 2008
Deviations:
no
Qualifier:
according to
Guideline:
EPA OPPTS 870.3700 (Prenatal Developmental Toxicity Study)
Version / remarks:
August 1998
Deviations:
no
GLP compliance:
yes (incl. certificate)
Remarks:
dd 3 November 2015
Limit test:
no

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
liquid
Details on test material:
- Lot/batch No.: 890000543374
- Appearance: Clear light yellow liquid
- Test item storage: At room temperature container flushed with nitrogen
- Purity/composition correction factor: No correction factor required
- Expiration date of the lot/batch: 10 February 2019
- Density: 0.8518 g/cm3 at 20°C
- Purity test date: 2016-02-16
- Analytical purity: 93.6%
- Purity/composition correction factor: No correction factor required
- Test item handling: No specific handling conditions required
- Solubility in water: Dispersible
- Stability in water: Stability for at least 6 hours at room temperature under normal laboratory light conditions is confirmed over the concentration range 0.5 - 5 mg/mL
Specific details on test material used for the study:
STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: At room temperature container flushed with nitrogen
- Stability under test conditions: stable
- Solubility and stability of the test substance in the solvent/vehicle: Stability for at least 6 hours at room temperature under normal laboratory light conditions is confirmed over the concentration range 0.5 – 5 mg/mL, Test Facility Study No. 498018.

Test animals

Species:
rat
Strain:
other: Crl:WI(Han)
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Charles River Deutschland, Sulzfeld, Germany
- Age at study initiation: 10-14 weeks
- Weight at study initiation: 171-247 g
- Fasting period before study: no
- Housing: individually housed in Macrolon plastic cages (MIII type, height 18 cm). Sterilized sawdust as bedding material (Lignocel S 8-15, JRS - J.Rettenmaier & Söhne GmbH + CO. KG, Rosenberg, Germany) and paper as cage enrichment/nesting material (Enviro-dri, Wm. Lillico & Son (Wonham Mill Ltd), Surrey, United Kingdom) were supplied.
- Diet: Free access to pelleted rodent diet (SM R/M-Z from SSNIFF® Spezialdiäten GmbH, Soest, Germany).
- Water: free access to tap water.
- Acclimation period: At least 5 days prior to treatment.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21.7-22.4
- Humidity (%): 45.9-69.4
- Air changes (per hr): at least 10
- Photoperiod (hrs dark / hrs light): 12/12

IN-LIFE DATES: From: 28 June 2016 To: 21 July 2016

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
water
Details on exposure:
PREPARATION OF DOSING SOLUTIONS:
Formulations (w/w) were prepared daily within 6 hours prior to dosing and were homogenized to a visually acceptable level. No adjustment was made for specific gravity/density of the vehicle. Adjustment was made for specific gravity of the test item (0.8518 g/cm3 at 20°C). No correction was made for the purity/composition of the test item.

VEHICLE
- Justification for use and choice of vehicle: Based on trial formulations performed at Charles River Den Bosch and on information provided by the Sponsor.
- Concentration in vehicle: 0.8, 2.5 and 7.5 mg/mL
- Amount of vehicle (if gavage): 10 mL/kg bw
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
Analyses were conducted on a single occasion during the treatment phase (05 July 2016), according to a validated method (Test Facility Study No. 498027). Samples of formulations were analyzed for homogeneity (highest and lowest concentration) and accuracy of preparation (all concentrations). Stability in vehicle over 6 hours at room temperature under normal laboratory light conditions was also determined (highest concentration only).
The accuracy of preparation was considered acceptable if the mean measured concentrations were 85-115% of the target concentration. Homogeneity was demonstrated if the coefficient of variation was ≤ 10%. Formulations were considered stable if the relative difference before and after storage was maximally 10%.
Details on mating procedure:
Untreated females were mated at the Supplier, and were at Day 0 or 1 post-coitum on arrival at the Test Facility (Day 0 post-coitum was the day of successful mating; confirmed by vaginal plug).
Duration of treatment / exposure:
From Days 6 to 20 post-coitum, inclusive
Frequency of treatment:
Once daily
Duration of test:
15 days
Doses / concentrationsopen allclose all
Dose / conc.:
0 mg/kg bw/day
Remarks:
Vehicle controls
Dose / conc.:
8 mg/kg bw/day
Dose / conc.:
25 mg/kg bw/day
Dose / conc.:
75 mg/kg bw/day
No. of animals per sex per dose:
22 females/dose
Control animals:
yes, concurrent vehicle
Details on study design:
- Dose selection rationale: Dose levels were selected based on the results of a combined 28-day/repro-screening study (OECD 422, Test Facility Study No. 498018), in which dose levels of 0, 5, 15 and 50 mg/kg were tested. At 50 mg/kg, hunched posture (both sexes) and lower motor activity (males only) were noted. Clinical biochemistry changes (lower total protein level, higher sodium and inorganic phosphate level) were observed in males at 50 mg/kg. Moreover, rats treated at 50 mg/kg showed treatment-related findings in the stomach, consisting of diffuse lymphogranulocytic inflammation of the forestomach (males only) and hyperplasia of the forestomach (both sexes). These findings were considered not to be adverse in toxicological terms. No reproduction or developmental toxicity was observed by treatment up to 50 mg/kg. In the previous dose range finding study (Test Facility Study No. 498017), rats treated at 150 mg/kg for 14 days showed significant clinical signs, weight loss, reduced food intake, lower thymus and spleen weights and 1/3 female was killed in extremis.

Examinations

Maternal examinations:
CAGE SIDE OBSERVATIONS: Yes
- Time schedule: at least twice daily for mortality and viability, at least once daily from day 2 post-coitum onwards up to the day prior to necropsy for clinical signs.

BODY WEIGHT: Yes
- Time schedule for examinations: Days 2, 6, 9, 12, 15, 18 and 21 post-coitum.

FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study): Yes
- Time schedule for examinations: Days 2-6, 6-9, 9-12, 12-15, 15-18 and 18-21 post-coitum

WATER CONSUMPTION AND COMPOUND INTAKE (if drinking water study): Subjective appraisal was maintained during the study, but no quantitative investigation was introduced as no treatment related effect was suspected.

POST-MORTEM EXAMINATIONS: Yes
- Sacrifice on gestation day # 21
- Organs examined: each ovary and uterine horn, stomach (10 females/group in groups).

Ovaries and uterine content:
The ovaries and uterine content was examined after termination: Yes
Examinations included:
- Gravid uterus weight: Yes
- Number of corpora lutea: Yes
- Number of implantations: Yes
- Number of early resorptions: Yes
- Number of late resorptions: Yes
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]
Statistics:
The following statistical methods were used to analyze the data:
• If the variables could be assumed to follow a normal distribution, the Dunnett-test (many-to-one t-test) based on a pooled variance estimate was applied for the comparison of the treated groups and the control group.
The Steel-test (many-to-one rank test) was applied if the data could not be assumed to follow a normal distribution.
• The Fisher Exact-test was applied to frequency data.
• The Mann Whitney test was used to compare mean litter proportions (percent of litter) of the number of viable and dead fetuses, early and late resorptions, total resorptions, pre- and post-implantation loss, and sex distribution.
• Mean litter proportions (percent per litter) of total fetal malformations and developmental variations (external, visceral and skeletal), and each particular external, visceral and skeletal malformation or variation were subjected to the Kruskal-Wallis nonparametric ANOVA test to determine intergroup differences. If the ANOVA revealed statistically significant (p<0.05) intergroup variance, Dunn’s test was used to compare the compound-treated groups to the control group.
All tests were two-sided and in all cases p < 0.05 was accepted as the lowest level of significance. Group means were calculated for continuous data and medians were calculated for discrete data (scores) in the summary tables. Test statistics were calculated on the basis of exact values for means and pooled variances. Individual values, means and standard deviations might be rounded off before printing. Therefore, two groups might display the same printed means for a given parameter, yet display different test statistics values.
No statistics were applied for data on maternal survival, pregnancy status, group mean numbers of dead fetuses, early and late resorptions, and pre- and post-implantation loss.
Indices:
For each litter the following calculations were performed:
Pre-implantation loss (%) = [(number of corpora lutea - number of implantation sites)/number of corpora lutea] x 100
Post-implantation loss (%) = [(number of implantation sites - number of live fetuses)/number of implantation sites] x 100
Viable fetuses affected/litter (%) = [(number of viable fetuses affected/litter)/(number of viable fetuses/litter)] x 100
Historical control data:
Historical control data from the same testing lab from 2014-2015 are available.

Results and discussion

Results: maternal animals

General toxicity (maternal animals)

Clinical signs:
effects observed, treatment-related
Description (incidence and severity):
No toxicologically relevant clinical signs were noted by treatment up to 75 mg/kg.
Rales was observed for three females at 75 mg/kg (nos. 74, 79 and 80) for 1-3 days. As this finding was considered to be related to local stomach effects observed in some females at 75 mg/kg (see sections 7.2.5 and 7.2.6), this was probably related to treatment with test item.
Incidental findings noted included alopecia, scabs and salivation. These findings were not considered to be test item related as they occurred in single females, in absence of a dose-related trend and within the range of background findings to be expected for rats of this age and strain treated under the conditions in this study.
Mortality:
no mortality observed
Body weight and weight changes:
effects observed, treatment-related
Description (incidence and severity):
At 75 mg/kg, mean body weights were lower than controls from Day 9 post-coitum onwards, reaching statistical significance on Day 15 and 21 post-coitum. Mean body weight gains were statistically significantly lower at 75 mg/kg from Day 9 post-coitum onwards, when compared to controls. In addition, weight gain corrected for gravid uterus was statistically significantly lower at 75 mg/kg when compared to controls (5.4 gram versus 26.5 gram).
Mean body weights and (corrected) body weight gains at 8 and 25 mg/kg remained in the same range as controls over the treatment period.
Food consumption and compound intake (if feeding study):
effects observed, treatment-related
Description (incidence and severity):
Absolute and relative food consumption were statistically significantly lower at 75 mg/kg during the entire treatment period.
Food consumption was considered to be unaffected by treatment up to 25 mg/kg.
Food efficiency:
not examined
Water consumption and compound intake (if drinking water study):
no effects observed
Description (incidence and severity):
Only subjective appraisal was made, as no treatment-related effect was suspected.
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, treatment-related
Description (incidence and severity):
Three females at 75 mg/kg (nos. 70, 74 and 77) showed an irregular surface of the forestomach, which was considered to be test item related. The microscopic correlate was diffuse hyperplasia of the forestomach and/or hyperkeratosis of the limiting ridge. Moreover, thymus reduced in size and Gi-tractus distended with gas were noted for single females at 75 mg/kg (nos. 70 and 75, respectively).
No macroscopic findings were noted in the control, 8 and 25 mg/kg groups.
Neuropathological findings:
not examined
Histopathological findings: non-neoplastic:
effects observed, treatment-related
Description (incidence and severity):
Test item-related microscopic findings noted for the stomach of the 75 mg/kg group females consisted of:
• Ulceration of the forestomach (marked) in 1/10 females.
• Diffuse hyperplasia of the forestomach in 2/10 females (minimal and slight).
• Hyperkeratosis of the limiting ridge at increased incidence and/or severity (3/10 minimal, 2/10 slight) compared to 1/10 female at 25 mg/kg at minimal degree which was regarded to be within background levels.
These stomach findings were considered to be most likely a local response to exposure with the test item, whereby the marked ulceration can be regarded as adverse.
There were no other test item-related histologic changes. Remaining histologic stomach changes were considered to be incidental findings. There was no test item related alteration in the prevalence, severity, or histologic character of those incidental tissue alterations.
Histopathological findings: neoplastic:
no effects observed
Other effects:
no effects observed

Maternal developmental toxicity

Number of abortions:
no effects observed
Description (incidence and severity):
There were no toxicologically relevant effects by treatment up to 75 mg/kg.
One Group 3 female (no. 45) had resorptions only. All other females (with the exception onf one control female and one group 2 female that were not pregnant) were pregnant and had litters with viable fetuses.
Pre- and post-implantation loss:
no effects observed
Description (incidence and severity):
There were no toxicologically relevant effects on the numbers of pre- or post-implantation loss by treatment up to 75 mg/kg.
The number of pre-implantation loss was slightly higher in the 75 mg/kg group when compared to controls. This change was not statistically significant and as treatment started from implantation onwards (i.e. Day 6 post-coitum), this was not considered to be treatment related.
Total litter losses by resorption:
no effects observed
Description (incidence and severity):
There were no toxicologically relevant effects by treatment up to 75 mg/kg.
Early or late resorptions:
no effects observed
Description (incidence and severity):
There were no toxicologically relevant effects by treatment up to 75 mg/kg.
Dead fetuses:
no effects observed
Description (incidence and severity):
There were no effects by treatment up to 75 mg/kg.
Changes in pregnancy duration:
not examined
Description (incidence and severity):
Migrated Data from removed field(s)
Field "Effects on pregnancy duration" (Path: ENDPOINT_STUDY_RECORD.DevelopmentalToxicityTeratogenicity.ResultsAndDiscussion.ResultsMaternalAnimals.MaternalDevelopmentalToxicity.EffectsOnPregnancyDuration): not examined
Changes in number of pregnant:
no effects observed
Description (incidence and severity):
One control female (no. 5) and one Group 2 female (no. 30) were not pregnant. One Group 3 female (no. 45) had resorptions only. All other females were pregnant and had litters with viable fetuses.

Effect levels (maternal animals)

open allclose all
Key result
Dose descriptor:
NOAEL
Remarks:
systemic effects
Effect level:
25 mg/kg bw/day
Based on:
test mat.
Basis for effect level:
body weight and weight gain
food consumption and compound intake
histopathology: non-neoplastic
Key result
Dose descriptor:
NOAEL
Remarks:
local effects
Effect level:
25 mg/kg bw/day
Based on:
test mat.
Basis for effect level:
gross pathology
histopathology: non-neoplastic

Maternal abnormalities

Key result
Abnormalities:
no effects observed

Results (fetuses)

Fetal body weight changes:
effects observed, non-treatment-related
Description (incidence and severity):
Mean combined (male and female) body weights were 5.3, 5.2, 5.4 and 5.1 gram for the control, 8, 25 and 75 mg/kg groups, respectively.
Fetal body weights (both sexes) were statistically significantly lower at 75 mg/kg when compared to controls. No clear dose related trend was observed and the values at 75 mg/kg were within or only slightly below the range of available historical control data. The level of statistical significance was considered to have arisen as a result of the slightly high concurrent control values, when compared to the available historical control data.
Migrated Data from removed field(s)
Field "Fetal/pup body weight changes" (Path: ENDPOINT_STUDY_RECORD.DevelopmentalToxicityTeratogenicity.ResultsAndDiscussion.ResultsFetuses.FetalPupBodyWeightChanges): not examined
Reduction in number of live offspring:
no effects observed
Description (incidence and severity):
No treatment-related effects were observed up to the dose level of 75 mg/kg bw/day.
Changes in sex ratio:
no effects observed
Description (incidence and severity):
The male:female sex ratio was unaffected by treatment up to 75 mg/kg.
Mean sex ratios (males:females) were 51:49, 50:50, 52:48 and 52:48 for the control, 8, 25 and 75 mg/kg groups, respectively.
Changes in litter size and weights:
no effects observed
Description (incidence and severity):
There were no treatment-related effects on litter size for any group.
Mean litter sizes were 10.2, 11.1, 10.2 and 10.6 fetuses/litter for the control, 8, 25 and 75 mg/kg groups, respectively.
Changes in postnatal survival:
not examined
External malformations:
effects observed, non-treatment-related
Description (incidence and severity):
There were no treatment related effects on external morphology following treatment up to 75 mg/kg.
The only malformation observed in groups treated with the test item was a small lower jaw that occurred in one fetus per group at 25 and 75 mg/kg bw/day. Both cases were confirmed skeletally, whereby the mandibles appeared to be fused along the mandibular symphysis. As these cases occurred singly and a small or absent lower jaw was seen previously (skeletally) in historical control data, they were considered chance findings.
Besides the noted jaw findings, one control fetus (A009-04) had a meningocele.
External variations were not seen in any group.
Skeletal malformations:
effects observed, non-treatment-related
Description (incidence and severity):
There were no treatment related effects on skeletal morphology following treatment up to 75 mg/kg.
The only skeletal malformation observed in this study was fused mandibles. This finding was noted for three fetuses, one per group at 8, 25 and 75 mg/kg bw/day. The same fetuses at 8 and 75 mg/kg bw/day had a small lower jaw as well, and the fetus at 25 mg/kg bw/day had only one socket. These findings occurred singly and do not indicate a treatment relationship.
The variation of slightly to moderately malaligned sternebrae was statistically significantly lower at 25 mg/kg bw/day. The incidence of this finding was 27.7%, 35.8%, 16.7% and 36.9% per litter in controls, 8, 25 and 75 mg/kg bw/day groups, respectively. The concurrent control value was for unknown reasons higher than the available historical control upper limit, while the Group 3 (25 mg/kg bw/day) value was within the applicable data range (4.4%-21.3% per litter). As no dose response could be established for this finding and the occurrence of fewer malaligned sternebrae is not adverse, the lower incidence in Group 3 was not considered to be toxicologically relevant.
Other skeletal variations that were noted occurred in the absence of a dose-related incidence trend, infrequently and/or at frequencies that were within the range of available historical control data. Therefore, they were not considered treatment related.
Visceral malformations:
effects observed, non-treatment-related
Description (incidence and severity):
There were no treatment related effects on visceral morphology following treatment up to 75 mg/kg.
Visceral malformations were only noted for one Group 2 and two Group 3 fetuses. No visceral malformations were noted in the control and highest dose group. In Group 3, a situs inversus was noticed in one fetus during eviscerating, and in another fetus a small left eye was found at soft tissue cephalic examination. In Group 2, the fetus with a small lower jaw had no urinary bladder. Due to the single occurrence and/or as they were noted in historical control fetuses (i.e. small eyes and situs inversus), all three malformations were considered to be chance findings.
Of the visceral variations, the finding of small supernumerary liver lobes was most common. It was observed in 4.5%, 4.4%, 6.7% and 8.6% of fetuses per litter in Groups 1, 2, 3 and 4, respectively. The mean litter incidence of Group 4 was slightly above the maximum historical control value (7.7% per litter), but as it was not statistically significantly higher when compared to the concurrent control group and this variation is most common in historical controls, this was not considered to be treatment related.
The other variations that were noted in this study occurred singly or at low incidences in the absence of a dose-related incidence trend and therefore were not considered to be treatment related.

Effect levels (fetuses)

Key result
Dose descriptor:
NOAEL
Effect level:
75 mg/kg bw/day
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: no adverse effects at the highest tested dose

Fetal abnormalities

Key result
Abnormalities:
no effects observed

Overall developmental toxicity

Key result
Developmental effects observed:
no

Any other information on results incl. tables

Accuracy of preparations and homogeinity:

The concentrations analysed in the formulations of Group 2, Group 3 and Group 4 were in agreement with target concentrations (mean accuracies 93, 94 and 90% with the coefficient of variation 1.6%, not determined and 3.6%, respectively; n = 6 for groups 2 and 4 and 2 for group 3).

Stability:

Analysis of the Group 4 formulation after storage yielded a relative difference of 15%. The value was higher than the criterion range of 10%. The accuracies of the samples after storage were 103% and 104%. The relatively high difference was obtained since the mean accuracy at t=0 was somewhat lower (i.e. 90%). Both at t=0 and t=6 hours the results were in the range of 85-115%. It demonstrated that the formulation of Group 4 set at level 7.51 mg/g could be used for adequately dosing in the storage period of 6 hours. The value of 15% indicated that degradation of the test item had not occurred during storage. Based on this, the formulation was found to be stable during storage at room temperature for at least 6 hours.

Table 1. Summary of the developmental effects observed in the study

Dose level (mg/kg bw/day)

0

8

25

75

Pregnant/total dams

21/22

21/22

22/22

22/22

-early resorptions

-late resorptions

(% per litter)

8.6

0.0

6.1

0.0

8.5

0.0

5.1

0.0

Dams with abortion, early deliveries, stillbirths, resorptions only and/or dead fetuses only

0

0

1

0

Pre-implantation loss (number and percent)

14 (5.1%)

11 (3.9%)

22 (7.6%)

28 (9.6%)

Post-implantation loss (number and percent)

21 (8.6%)

14 (6.1%)

12 (8.5%)

12 (5.1%)

Body weight on day 21

316

322

320

294*

Body weight gain day 6-21 (%)

47

49

48

35**

Gravid uterine weight (g)

73.8

77.4

73.8

71.4

Mean live offspring (number)

10.2

11.1

10.2

10.6

Live offspring (percent)

91.4

93.9

91.5

94.9

Mean fetal body weight males (g)

5.5

5.4

5.5

5.2*

Mean fetal body weight females

5.2

5.1

5.3

4.9*

Mean fetal body weight (sexes combined)

5.3

5.2

5.4

5.1*

Malformations (including runts) number and percent of fetuses per litter

 

 

1 (0.5%)

 

 

1 (0.3%)

 

 

2 (1.7%)

 

 

1 (0.4%)

Variations (% per litter)

-external

-soft tissue

-skeletal

 

0

9.9

76.6

 

0

9.4

77.1

 

0

12.6

69.0

 

0

11.6

77.9

*: significantly different from the control group at 0.05

**: significantly different from the control group at 0.01

Table 2. Historical control data

3-(isodecyloxy)-1-propanamine

 

 

 

 

 

 

Project 511810

 

 

 

 

 

 

 

 

Historical Control Data Rat: Crl:WI(Han) (outbred, SPF-Quality)

 

 

 

 

 

 

 

Gestation Day 21

 

Mean of Study Means

 

 

 

 

 

Study Date Range: 2014 - 2015

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Endpoint

Total

Mean

SD

Median

Min

Max

P5

P95

No of Studies

13

 

 

 

 

 

 

 

Total No. of Animals in the Control Group

304

 

 

 

 

 

 

 

No. of Animals that Died

0

 

 

 

 

 

 

 

No. of Animals that were Euthanized

0

 

 

 

 

 

 

 

No. of Animals that Aborted or Delivered

3

 

 

 

 

 

 

 

Percent Pregnant

 

98.8

2.73

100.0

90.9

100.0

97.1

100.0

No. of Animals Examined at Laparohysterectomy

301

 

 

 

 

 

 

 

No. Nongravid

4

 

 

 

 

 

 

 

No. Gravid

297

 

 

 

 

 

 

 

No. with Only Resorptions

2

 

 

 

 

 

 

 

No. of Dams with Live Fetuses

295

 

 

 

 

 

 

 

Mean No. Viable Fetuses/Dam

 

10.7

0.71

10.6

9.1

11.6

10.3

11.2

Total No. Viable Fetuses

3194

 

 

 

 

 

 

 

Viable Fetuses (%/Litter)

 

95.2

2.63

95.9

88.9

98.4

93.6

96.8

Mean No. Postimplantation Loss/Dam

 

0.5

0.15

0.4

0.2

0.7

0.4

0.6

Total No. Postimplantation Losses

134

 

 

 

 

 

 

 

Postimplantation Loss (%/Litter)

 

4.8

2.63

4.1

1.6

11.1

3.2

6.4

Dead Fetuses (%/Litter)

 

0.0

0.11

0.0

0.0

0.4

0.0

0.1

Early Resorptions (%/Litter)

 

4.7

2.62

4.1

1.6

11.1

3.2

6.3

Late Resorptions (%/Litter)

 

0.0

0.11

0.0

0.0

0.4

0.0

0.1

Mean No. Implantations/Dam

 

11.2

0.69

11.1

9.6

12.0

10.8

11.6

Mean No. Corpora Lutea/Dam

 

11.9

0.71

11.7

10.9

13.2

11.5

12.3

Mean No. Preimplantation Loss/Dam

 

0.7

0.32

0.6

0.2

1.3

0.5

0.9

Total No. Preimplantation Losses

207

 

 

 

 

 

 

 

Preimplantation Loss (%/Litter)

 

6.2

3.43

5.8

2.0

14.5

4.2

8.3

Total No. Male Fetuses

1617

 

 

 

 

 

 

 

Total No. Female Fetuses

1577

 

 

 

 

 

 

 

% Males/Litter

 

50.8

2.12

50.7

46.6

53.7

49.5

52.0

% Female/Litter

 

49.2

2.12

49.3

46.3

53.4

48.0

50.5

Mean Fetal Body Weight (g)

 

5.2

0.08

5.2

5.1

5.3

5.1

5.2

Mean Male Body Weight (g)

 

5.4

0.10

5.4

5.2

5.5

5.3

5.4

Mean Female Body Weight (g)

 

5.1

0.07

5.1

5.0

5.2

5.0

5.1

Mean Male Placenta Weight (g)1

 

0.46

0.01

0.47

0.44

0.47

0.4

0.5

Mean Female Placenta Weight (g)1

 

0.44

0.01

0.44

0.42

0.45

0.4

0.5

1Based on 4 datasets

 

 

 

 

 

 

 

 

3-(isodecyloxy)-1-propanamine

 

 

 

 

 

 

 

Project 511810

 

 

 

 

 

 

 

 

 

Historical Control Data Rat: Crl:WI(Han) (outbred, SPF-Quality)

 

 

 

 

 

 

 

 

 

Gestation Day 21

 

 

 

 

 

 

 

 

 

Study Date Range: 2014 - 2015

 

 

 

 

 

 

 

 

 

No. of Studies

13

 

 

 

 

 

 

 

 

Total No. Fetuses/Litters Examined Externally

3194

295

 

 

 

 

 

 

 

Total No. Fetuses/Litters Examined Viscerally

2061

295

 

 

 

 

 

 

 

Total No. Fetuses/Litters Examined Skeletally

2059

295

 

 

 

 

 

 

 

 

Mean of Study Means

 

 

 

 

 

Summary Incidence

 

(% Per Litter Basis)

 

 

 

 

 

(Total No. Affected)

MALFORMATIONS

Mean

SD

Median

Min

Max

P5

P95

Fetuses

Litters

Total External Malformations

 

 

 

 

 

 

 

1

1

Total Visceral Malformations

 

 

 

 

 

 

 

7

7

Total Skeletal Malformations

 

 

 

 

 

 

 

15

15

Total Malformations

 

 

 

 

 

 

 

22

22

EXTERNAL

 

 

 

 

 

 

 

 

 

Exencephaly

0.0

0.14

0.0

0.0

0.5

0.0

0.1

1

1

Eye(s)- Open

0.0

0.14

0.0

0.0

0.5

0.0

0.1

1

1

VISCERAL

 

 

 

 

 

 

 

 

 

Diaphragmatic Hernia

0.0

0.08

0.0

0.0

0.3

0.0

0.1

1

1

Eye(s)- Absent and/or Small

0.1

0.26

0.0

0.0

0.9

0.0

0.2

3

3

Hydrocephaly- External

0.0

0.12

0.0

0.0

0.5

0.0

0.1

1

1

Situs Inversus

0.2

0.34

0.0

0.0

1.0

0.0

0.4

3

3

SKELETAL

 

 

 

 

 

 

 

 

 

Jaw- Upper Jaw Small

0.1

0.22

0.0

0.0

0.8

0.0

0.2

1

1

Jaw- Lower Jaw Absent or Small

0.1

0.22

0.0

0.0

0.8

0.0

0.2

1

1

Limb Bone(s)- Bent

0.3

0.44

0.0

0.0

1.1

0.0

0.5

4

4

Rib Anomaly

0.1

0.31

0.0

0.0

1.1

0.0

0.3

1

1

Skull Anomaly

0.1

0.34

0.0

0.0

1.2

0.0

0.3

2

2

Sternebra(e)- Fused

0.1

0.29

0.0

0.0

1.0

0.0

0.3

2

2

Sternebra(e) Malaligned (Severe)

0.0

0.08

0.0

0.0

0.3

0.0

0.1

1

1

Sternoschisis

0.1

0.22

0.0

0.0

0.8

0.0

0.2

1

1

Vertebral Anomaly With or Without Associated Rib Anomaly

0.2

0.53

0.0

0.0

1.9

0.0

0.5

3

3

Vertebral Centra Anomaly

0.1

0.22

0.0

0.0

0.8

0.0

0.2

1

1

3-(isodecyloxy)-1-propanamine

 

 

 

 

 

 

 

Project 511810

APPENDIX 5

 

 

 

 

 

 

 

 

 

Historical Control Data Rat: Crl:WI(Han) (outbred. SPF-Quality)

 

 

 

 

 

 

 

 

 

Gestation Day 21

 

 

 

 

 

 

 

 

 

 

Mean of Study Means

 

 

 

 

Summary Incidence

 

(% Per Litter Basis)

 

 

 

 

 

(Total No. Affected)

VARIATIONS

Mean

SD

Median

Min

Max

P5

P95

Fetuses

Litters

EXTERNAL

 

 

 

 

 

 

 

 

 

None Observed

 

 

 

 

 

 

 

 

 

VISCERAL

 

 

 

 

 

 

 

 

 

Kidney(s)- Renal Papilla(e) Absent and/or Small

0.1

0.25

0.0

0.0

0.9

0.0

0.2

2

2

Liver- Appendix

1.2

0.56

1.3

0.3

2.3

0.9

1.6

23

21

Liver- Discolored

0.1

0.30

0.0

0.0

1.0

0.0

0.3

3

3

Liver- Small Supernumerary Lobe(s)

4.0

1.96

4.0

1.3

7.7

2.8

5.2

69

58

Spleen- Supernumerary

0.0

0.14

0.0

0.0

0.5

0.0

0.1

1

1

Thymus- Partially Undescended Horn(s)

1.3

1.55

0.8

0.0

4.3

0.3

2.2

34

23

Thyroid- Discolored

0.1

0.36

0.0

0.0

1.3

0.0

0.3

1

1

Ureter(s)- Convoluted

1.0

2.39

0.0

0.0

8.7

0.0

2.5

43

28

Ureter(s)- Dilated

0.9

2.33

0.0

0.0

8.5

0.0

2.3

44

19

SKELETAL

 

 

 

 

 

 

 

 

 

7th Cervical Rudimentary Rib(s)

1.7

1.34

1.2

0.0

4.4

0.9

2.5

30

26

7th Cervical Full Rib(s)

0.1

0.36

0.0

0.0

1.1

0.0

0.4

2

2

14th Full Rib(s)

5.7

4.65

5.2

0.0

13.1

2.9

8.5

88

64

14th Rudimentary Rib(s)

44.1

19.84

54.4

19.0

72.0

32.1

56.1

798

250

Metacarpal(s) and/or Metatarsal(s) Unossified

2.2

1.97

1.0

0.0

6.3

1.0

3.4

41

24

Pelvic Girdle- Caudal Shift

6.6

3.77

7.1

1.7

12.8

4.3

8.9

127

71

Rib(s)- Bent

10.6

7.78

10.2

0.8

22.3

5.9

15.3

162

85

Rib(s)- Short

0.0

0.06

0.0

0.0

0.2

0.0

0.0

1

1

Skull- Reduced Ossification

2.7

2.55

1.8

0.0

7.0

1.2

4.3

81

46

Skull- Supernumerary Site

0.0

0.14

0.0

0.0

0.5

0.0

0.1

1

1

Sternebra(e) #1, #2, #3 and/or #4 Unossified

0.2

0.31

0.0

0.0

0.8

0.0

0.3

3

3

Sternebra(e) #5 and/or #6 Unossified

0.9

1.33

0.0

0.0

4.1

0.1

1.7

37

23

Sternebrae- Malaligned (Slight or Moderate)

11.1

5.72

8.9

4.4

21.3

7.6

14.5

188

131

Sternum- Supernumerary Ossification Site

0.1

0.31

0.0

0.0

1.1

0.0

0.3

1

1

Vertebral Centra- Reduced Ossification

0.6

0.88

0.4

0.0

3.0

0.1

1.2

12

12

Applicant's summary and conclusion

Conclusions:
In a GLP-compliant guideline study, pregnant Crl:WI(Han) rats were dosed the test substance by oral gavage at 0 (vehicle controls), 8, 25 and 75 mg/kg bw/day from Day 6 to Day 21 post-coitum. The NOAEL for maternal toxicity was set at 25 mg/kg bw/day based on reduced body weight and body weight gain, reduced consumption and local irritation effects in the stomach (ulceration, diffuse forestomach hyperplasia and/or increased incidence and/or severity of hyperkeratosis of the limiting ridge) at the highest dose level. The NOAEL for developmental effects was set at 75 mg/kg bw/day, based on the absence of adverse effects at the highest dose level.
Executive summary:

In a GLP-compliant OECD Guideline 414 study, pregnant rats (22/dose) were administered the test substance solution in water by oral gavage at 0 (vehicle controls), 8, 25 and 75 mg/kg bw/day from Day 6 to Day 21 post-coitum. All animals surviving to Day 21 post-coitum were subjected to an examination post-mortem and external, thoracic and abdominal macroscopic findings were recorded. The stomach and gross lesions were collected and fixed from all animals at necropsy.

Histopathological examination was performed on the stomach from 10 selected females of all groups.

 

Treatment at 75 mg/kg resulted in statistically significantly lower body weights and body weight gains on Days 15 and 21 post-coitum and from Day 9 post-coitum onwards, respectively. In addition, weight gain corrected for gravid uterus was statistically significantly lower at 75 mg/kg bw/day when compared to controls (5.4 gram versus 26.5 gram). Absolute and relative food consumption were statistically significantly lower at 75 mg/kg bw/day during the entire treatment period. Test item-related morphological findings were present in the stomach of females treated at 75 mg/kg bw/day, consisting of ulceration, diffuse forestomach hyperplasia and/or increased incidence and/or severity of hyperkeratosis of the limiting ridge. Based on these effects the NOAELs for maternal toxicity for both systemic and local effects were set to 25 mg/kg bw/day.

There were no effects on the number of abortions, early or late resorptions, total implantation losses or number of live fetuses. There were no toxicologically relevant or test item related external, visceral and skeletal malformations or variations noted by treatment up to 75 mg/kg. Fetal body weights in both sexes were statistically significantly lower at 75 mg/kg bw/day when compared to controls. The level of statistical significance was considered to have arisen as a result of the slightly high concurrent control value. In addition, as no clear dose related trend was observed and no effects on skeletal developmental were noted, this was considered to be related to the significant lower body weights of the dams rather than a developmental cause. Hence, this minimal change in fetal body weight was considered to be non-adverse.

Based on this the NOAEL for developmental effects was set at 75 mg/kg bw/day.