Registration Dossier

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Diss Factsheets

Administrative data

Key value for chemical safety assessment

Effects on fertility

Description of key information

No reproductive/fertility toxicity study is available on the substance hydroxybenzomorpholine. Two repeated dose toxicity studies (28-day and 90-day studies) via the oral route are available on hydroxybenzomorpholine (OECD TG 407 and OECD TG 408 respectively). Based on the results obtained in these two relevant and reliable key studies, the substance hydroxybenzomorpholine does not present effects for fertility toxicity via the oral route (absence of adverse effects on reproductive organs or tissues).


No Extended One- Generation Reproductive Toxicity Study is available for hydroxybenzomorpholine. According to Regulation (EC) No 1907/2006, Annex IX, 8.7.3, column 1, an Extended One- Generation Reproductive Toxicity Study (B.56 of the Commission Regulation on test methods as specified in Article 13(3) or OECD 443), basic test design (cohorts 1A and 1B without extension to include a F2 generation), one species, most appropriate route of administration, having regard to the likely route of human exposure, has to be performed if the available repeated dose toxicity studies (e.g. 28-day or 90-day studies, OECD 421 or 422 screening studies) indicate adverse effects on reproductive organs or tissues or reveal other concerns in relation with reproductive toxicity. Based on the results of the available repeated dose toxicity studies (OECD TG 407 and OECD TG 408) (i.e. absence of adverse effects on reproductive organs or tissue or any other concerns in relation with reproductive toxicity), an Extended One- Generation Reproductive Toxicity Study (B.56 of the Commission Regulation on test methods as specified in Article 13(3) or OECD 443), need not to be conducted.

Link to relevant study records
Reference
Endpoint:
screening for reproductive / developmental toxicity
Data waiving:
study scientifically not necessary / other information available
Justification for data waiving:
the study does not need to be conducted because a pre-natal developmental toxicity study is available
Effect on fertility: via oral route
Endpoint conclusion:
no study available
Effect on fertility: via inhalation route
Endpoint conclusion:
no study available
Effect on fertility: via dermal route
Endpoint conclusion:
no study available
Additional information

No reproductive/fertility toxicity data is available for hydroxybenzomorpholine. However, two repeated dose toxicity studies in the rat (28-day and 90-day studies) via the oral route are available (OECD TG 407 and OECD TG 408 respectively), which demonstrated the absence of adverse effects on reproductive organs or tissues.

In the 28-day study (Boutemy, similar to OECD TG 407, GLP, 1989, Klimisch 1) (detailed summary is provided in section 7.5 Repeated dose toxicity), hydroxybenzomorpholine, was administered by gavage once daily to groups of Sprague Dawley rats (10/sex) for 30 days (males) and 31 days (females). The dose levels were 10, 100 and 1000 mg/kg bw. The control group (10/sex) received the vehicle alone (hydrogel with 2 grams polysorbate 80 per 100 ml sterile water for injectable preparations). All animals were killed at the end of the study. Organ weights were recorded. Macroscopic and histopathologic examination of tissues was undertaken.

Examination of reproductive organ weights only showed changes in the high dose (1000 mg/kg/day dose with an increase in relative testes weight of 11 % in the males (no significant change in absolute weight was noted). The increase in testes weight noted in this group was supposed be related to the decrease in bodyweight (at the end of the study, a decrease in weight gain of the order of 7.5 %was noted in male animals treated with the 1000 mg/kg/day dose level).

In the 90-day study (Hopkins, OECD TG 408, GLP, 1996, Klimisch 1) (detailed summary is provided in section 7.5 Repeated dose toxicity), hydroxybenzomorpholine, was administered by gavage once daily to groups of rats of the Crl:CD BR strain (VAF plus) (10/sex) for 13 weeks. The dose levels were 5, 25 and 125 mg/kg/day. The control group (10/sex) received the vehicle alone (0.5% carboxymethylcellulose, with Tween 80 at 0.5% of the final formulation). At the end of the treatment period, all animals were killed by C02 asphyxiation and subjected to a macroscopic examination. Designated organs were weighed and a range of tissues preserved for microscopic examination.

There was no adverse effect on the bodyweight gain of males and females. There were no differences from control in organ weights (including reproductive organs: testes and ovaries) attributable to administration of hydroxybenzomorpholine. No abnormalities attributable to the test substance were noted during histopathological examination including reproductive tissues (epididymides, prostate, seminal vesicles, testes, mammary gland, ovaries, uterus and vagina).

Hence, based on the results from the two key repeated dose toxicity studies via the oral route, hydroxybenzomorpholine did not induce any relevant adverse effect on male and female reproductive organs or tissues.

Effects on developmental toxicity

Description of key information

In the key developmental toxicity study in the rat (Foulon, OECD TG 414, GLP, 2005, Klimisch 1), the NOAEL was concluded to be 125 mg/kg/day for maternal toxicity and 500 mg/kg/day for developmental toxicity (highest dose tested).

Link to relevant study records
Reference
Endpoint:
developmental toxicity
Type of information:
experimental study
Adequacy of study:
key study
Study period:
23 July 2004 to 12 April 2005
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to guideline
Guideline:
OECD Guideline 414 (Prenatal Developmental Toxicity Study)
Version / remarks:
22 January 2001
Deviations:
no
GLP compliance:
yes
Limit test:
no
Specific details on test material used for the study:
SOURCE OF TEST MATERIAL
- Source and lot/batch number of test material: 0508918
- Expiration date of the lot/batch: September 2005.
- Purity test date: 98.3%


STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: at +4°C, protected from light and under nitrogen gas
- Stability under storage conditions: not specified
- Stability under test conditions: Before preparation, the vehicle was degassed by sonication for at least 15 minutes, then saturated
with nitrogen gas and kept under nitrogen atmosphere for 15 minutes.
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 4, 25 and 100 mg/mL and then homogenized using a magnetic stirrer.
The test item dosage forms were prepared under nitrogen atmosphere and stored for up to 9 days at +4°C, protected from light (using a glass beaker covered with aluminium foil) and under nitrogen atmosphere until delivery.
- Solubility and stability of the test substance in the solvent/dispersant/vehicle/test medium: Before the start of treatment, the suitability of the proposed preparation procedure was confirmed by the analysis of concentration, homogeneity and stability of dosage forms prepared using this
procedure. These analyses were performed in CIT/Study No. 26976 AHS at concentrations spanning those used in the present study.
During the treatment period, the concentration of dosage forms prepared for use in the study was checked.
- concentration control : The concentration of samples taken from each control and test item dosage form, prepared for use on the first and the last day of the dosing period, was determined according to the analytical method previously validated in CIT/Study No. 26976 AHS.

Species:
rat
Strain:
Sprague-Dawley
Remarks:
Crl CD® (SD) IGS BR, Caesarian Obtained, Barrier Sustained-Virus Antibody Free, (COBS-VAF®).
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: Charles River Laboratories France, L’Arbresle, France
- Age at study initiation: at the beginning of the treatment period, the animals were 10-11 weeks old
- Weight at study initiation: mean body weight of 260 g (range: 220 g to 294 g). The females were sexually mature and primigravid
- Fasting period before study: no data
- Housing: The animal room was disinfected before the arrival of the animals and cleaned regularly thereafter. The animals were individually housed in suspended wire-mesh cages (43.0 x 21.5 x 18.0 cm). A metal tray, containing autoclaved sawdust (SICSA, Alfortville, France), was placed under each cage. The sawdust was changed at least once a week. The cages were placed in numerical order, vertically on the racks.
- Diet (e.g. ad libitum): The animals had free access to A04 C pelleted maintenance diet, batch Nos. 40518 and 40617 (SAFE, Villemoisson, Epinay-sur-Orge, France), which was distributed weekly
- Water (e.g. ad libitum): The animals had free access to bottles containing tap water (filtered with a 0.22 μm filter).
- Acclimation period: a period of 5 days acclimation to the conditions of the study preceded the beginning of the mating period.
- Monitoring of estrous cycle: during the week of mating, the estrous cycle stage was determined periodically from a fresh vaginal lavage (stained with methylene blue).

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 2°C
- Humidity (%): 50 ± 20%
- Air changes (per hr): about 12 cycles/hour of filtered, non-recycled air.
The corresponding instrumentation and equipment are checked and calibrated at regular intervals. The temperature and relative humidity are recorded continuously and the records are checked daily and filed.
- Photoperiod (hrs dark / hrs light): 12h/12h (7:00 - 19:00)

IN-LIFE DATES: From: 7 september 2004 To: 6 October 2004
Route of administration:
oral: gavage
Vehicle:
CMC (carboxymethyl cellulose)
Details on exposure:
PREPARATION OF DOSING SOLUTIONS:
Before preparation, the vehicle was degassed by sonication for at least 15 minutes, then saturated with nitrogen gas and kept under nitrogen atmosphere for 15 minutes.
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 4, 25 and 100 mg/mL and then homogenized using a magnetic stirrer.

VEHICLE
- Justification for use and choice of vehicle (if other than water): The vehicle was 0.5% carboxymethylcellulose in purified water prepared using:
. purified water, obtained by reverse osmosis using a Milli-Ro 8 plus apparatus (Millipore SA, Saint-Quentin en Yvelines, France),
. carboxymethylcellulose, batch No. 101K0185, supplied by Sigma (Saint-Quentin-Fallavier, France).
- Concentration in vehicle: 250, 400 and 550 mg/kg
- Amount of vehicle (if gavage): 4, 25 and 100 mg/mL
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
The concentration of samples taken from each control and test item dosage form, prepared for use on the first and the last day of the dosing period, was determined according to the analytical method previously validated in CIT/Study No. 26976 AHS.
Details on mating procedure:
- Mating: the females in proestrus were placed overnight in the home cage of singly housed stock males (one female with one male). The males were from the same strain and the same breeder.
Each morning following mating, rats with spermatozoa found in a vaginal smear or sperm plug in situ were considered as pregnant animals. The day where evidence of mating was found was designated as day 0 post-coitum (p.c.).
- Allocation to groups: before day 3 p.c., the animals were allocated to the groups, according to a stratification procedure based on body weight, recorded on day 0 p.c., to ensure a comparatively similar mean body weight among the groups. A larger number of animals than necessary were paired, to permit the selection and/or replacement of individuals before start of treatment. The data on the replaced animals are filed but not presented in the study report.
Duration of treatment / exposure:
Each animal was given the dosage form once a day, at approximately the same time, from day 6 to day 19 post-coitum, inclusive.
Frequency of treatment:
once a day
Dose / conc.:
0 mg/kg bw/day
Dose / conc.:
20 mg/kg bw/day
Dose / conc.:
125 mg/kg bw/day
Dose / conc.:
500 mg/kg bw/day
No. of animals per sex per dose:
24
Control animals:
yes, concurrent vehicle
Details on study design:
Rationale for dose-level selection in agreement with the Sponsor
The high dose-level in the present study was set at 500 mg/kg, based on the results of a preliminary study performed at 250, 400 and 550 mg/kg (CIT/Study No. 26999 RSR). In this study, a transient decrease in food consumption and a transient body weight loss were observed at 400 and 550 mg/kg. These body weight losses resulted in a slightly decreased overall body weight gain, when compared to controls (-10%, both dose-levels). Accordingly, the dose-level of 500 mg/kg was expected to be associated with sufficient maternal toxicity, and was selected as a high dose-level for the present study. Low and intermediate dose-levels were set at 20 and 125 mg/kg to assess the dose-relationship of the toxic effects.
Maternal examinations:
CAGE SIDE OBSERVATIONS: Yes
Each animal was checked for mortality or signs of morbidity:
. at least twice a day during treatment period,
. at least once a day on other days.

DETAILED CLINICAL OBSERVATIONS: Yes
From arrival, the animals were observed at least once a day as part of routine examinations.
From the start of the treatment period, each animal was observed at least 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 female was recorded on days 0, 3, 6, 9, 12, 15, 18 and 20 post-coitum.

FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study): Yes
- Food consumption : The quantity of food consumed by each female was recorded for the following intervals:
. days 0-3, 3-6, 6-9, 9-12, 12-15, 15-18 and 18-20 post-coitum.


WATER CONSUMPTION AND COMPOUND INTAKE (if drinking water study): No

POST-MORTEM EXAMINATIONS: Yes
- Sacrifice on gestation day : Hysterectomies
On day 20 p.c., females were sacrificed by inhalation of carbon dioxide gas followed by cervical dislocation, and were submitted for a macroscopic post-mortem examination of the principal thoracic and abdominal organs.
The weight of the gravid uterus of each pregnant female (with at least one live fetus) was recorded.

Ovaries and uterine content:
The ovaries and uterine content was examined after termination: Yes
Examinations included:
The ovaries and uterus of the females were examined to determine:
. number of corpora lutea,
. number and distribution of dead and live fetuses,
. number and distribution of early and late resorptions,
. number and distribution of uterine scars,
. number and distribution of implantation sites.

The following classification was used to record:
. uterine scar: uterine implantation without implant,
. early resorption: evidence of implant without recognizable embryo,
. late resorption: dead embryo or fetus with external degenerative changes,
. dead fetus: non live fetus with discernible digits.
Uterine horns without visible implantation sites were immersed in an aqueous solution of ammonium sulphide (Salewski) to reveal the presence of uterine scars.
A gross evaluation of placentas was also undertaken.
Fetal examinations:
These examinations were carried in all the litters (females with at least one live fetus).
The fetal findings were described according to the glossary of the International Federation of Teratology Societies (IFTS) and classified as malformations or variations (Wise, Chahoud):
. malformation refers to a permanent structural change which is likely to adversely affect the survival or health,
. variation refers to a change that occurs within the normal population under investigation and is unlikely to adversely affect the survival or health (this might include a delay in growth, or morphogenesis, that otherwise followed a normal pattern of development).

- External examinations: Yes - Each fetus was submitted for a detailed external examination, which included the observation of all visible structures, surfaces and orifices.
The fetuses were killed by a subcutaneous injection of thiopental sodium.
- Soft tissue examinations: Yes: Approximately half of the fetuses per litter were fixed with Harrisson’s fluid. A detailed soft tissue examination was performed according to a free-hand serial sectioning technique (Wilson technique), which included the observation of all the organs and structures of the head, neck, thorax and abdomen.
- Skeletal examinations: Yes: The remaining fetuses per litter were eviscerated and then fixed with ethyl alcohol. A detailed examination of the skeleton (bone) was performed after staining with alizarin red S (modified Dawson technique). This examination included the observation of all the bone structures of the head, spine, rib cage, pelvis and limbs.
- Head examinations: No
- Sex of fetuses: The sex of each fetus was determined at the time of evisceration or at the time of serial sectioning.
Statistics:
Mean values were compared by one-way analysis of variance and the Dunnett test (mean values being considered as normally distributed and variances being considered as homogeneous).
Percentage values were compared by the Fisher exact probability test.
Indices:
Pre- and post-implantation loss, fetal or litter incidence and mean proportion of affected fetuses. Photographs of fetuses were taken to document the findings and archived with the study files
Clinical signs:
no effects observed
Description (incidence and severity):
A satisfactory agreement was observed between the nominal and actual concentrations of the test item in the administered dosage forms analyzed, since the deviations from nominal concentration were within an acceptable range of ± 9%.
Clinical signs: All females given 500 mg/kg/day and the majority of females given 125 mg/kg/day had orange colored urine generally starting during early- or mid-gestation and lasting until the end of the study. This finding was considered to be evidence of systemic exposure following oral administration of test item. Six females given 500 mg/kg/day experienced excessive salivation, but this clinical sign was observed at a low frequency.
Mortality:
no mortality observed
Body weight and weight changes:
effects observed, treatment-related
Description (incidence and severity):
Net body weight change (body weight change over the dosing period, minus the gravid uterus weight) was statistically and significantly reduced by the test item treatment at 125 and 500 mg/kg/day (-19% p<0.05 and -52% p<0.001, respectively), when compared with the controls. The slight changes observed at 125 mg/kg/day were considered to bear no biological significance as there were no associated significant changes in body weight gain or carcass weight at this dose-level. Accordingly, there were no significant changes in body weight or body weight gain at 20 and 125 mg/kg/day.
Food consumption and compound intake (if feeding study):
effects observed, treatment-related
Description (incidence and severity):
When compared to controls, there was a moderate but statistically significant reduction in mean food consumption at 500 mg/kg/day throughout the dosing period. There were no changes in food intake at 20 and 125 mg/kg.
Number of abortions:
no effects observed
Pre- and post-implantation loss:
no effects observed
Total litter losses by resorption:
no effects observed
Early or late resorptions:
no effects observed
Dead fetuses:
no effects observed
Changes in pregnancy duration:
no effects observed
Changes in number of pregnant:
no effects observed
Other effects:
no effects observed
Description (incidence and severity):
At 500 mg/kg/day, the carcass weight, corresponding to maternal body weight on day 20 of gestation minus the gravid uterus weight, was lower (p<0.001, -7.4%) than the control group value.
There was no effect of treatment on the mean gravid uterus weight at any dose-level.
Details on maternal toxic effects:
The mean number of corpora lutea and implantations, and consequently the pre-implantation loss, were similar for all groups.
There was no effect of treatment on the group mean numbers of dead or live fetuses or on the extent of post-implantation losses.
There was also no effect of treatment on the percentage of male fetuses or on fetal body weight.
Key result
Dose descriptor:
NOAEL
Effect level:
ca. 125 mg/kg bw/day
Based on:
test mat.
Basis for effect level:
body weight and weight gain
Key result
Abnormalities:
no effects observed
Fetal body weight changes:
no effects observed
External malformations:
no effects observed
Description (incidence and severity):
Given their low incidence and distribution among groups, no external fetal malformations were considered to be related to treatment
Skeletal malformations:
effects observed, non-treatment-related
Description (incidence and severity):
There was a single fetus at 20 mg/kg showing various skeletal malformations. This isolated finding was considered to bear no relationship to treatment.
Visceral malformations:
no effects observed
Description (incidence and severity):
Given their low incidence and their distribution among groups, no soft tissue fetal malformations or variations were considered to be related to treatment. In particular, though there were 2/165, 3/159 and 1/161 fetuses showing a ventricular septal defect respectively at 20, 125 and 500 mg/kg, these slightly high incidences were considered to be fortuitous in the absence of a dose-relationship.
Description (incidence and severity):
Slightly high fetal and litter incidences of short 14th ribs (slightly above the upper limit of historical control data, fetal incidence range = 0.0 - 3.7%, litter incidence
range = 0.0 - 11.8%) were observed at 500 mg/kg, and a relationship to treatment with A025 could not be excluded. A slightly high fetal incidence of short 14th rib was also observed at 20 mg/kg, but since this fetal incidence was close to the upper limit of our historical control range, since litter incidence was similar to controls at this dose-level, and since there were no such changes at the higher dose-level of 125 mg/kg, this finding was considered to be unrelated to treatment with A025.
Key result
Dose descriptor:
NOAEL
Effect level:
ca. 500 mg/kg bw/day
Based on:
test mat.
Basis for effect level:
other:
Remarks on result:
other: no effects
Key result
Abnormalities:
no effects observed
Key result
Developmental effects observed:
no
Lowest effective dose / conc.:
500 mg/kg bw/day
Treatment related:
no


1.1       CHRONOLOGY OF THE STUDY

 

The chronology of the study is summarized as follows:


Procedure                                                                                                  Date

Study plan approved by:

.StudyDirector                                                                             17 December2003


.StudyMonitor                                                                              17 December2003

Mating of females(day 0post-coitum)

. First female (experimentalstartingdate)                                      7 September2004

.Last female                                                                                  16 September2004

 

First day of treatment(day 6post-coitum)

.First female                                                                                 13 September2004

.Last female                                                                                  22 September2004

 

Last day of treatment(day 19post-coitum)

.First female                                                                                 26 September2004

.Last female                                                                                        5 October2004

 

Hysterectomies(day 20post-coitum)

.First female                                                                                 27 September2004


. Last female (experimentalcompletiondate)                                    6 October2004

 

 

 

1.2   MATERNALDATA

The pregnancy status of the females is summarized in the table below:

 

Dose-level (mg/kg/day)

0

20

125

500

Number of mated females

24

24

24

24

Number of non-pregnant females

1

0

0

1

Number of females with live fetuses

23

24

24

23

 

 

 

Table: 1

 

 

 CLINICAL SIGNS (Summary table/Females/Pregnancy period)

 

 

Dose: (mg/kg/day)

0

20

125

500

 

MortalityFINALSACRIFICE

 

 

24

 

 

24

 

 

24

 

 

24

 

Secretion/ExcretionPTYALISM

 

 

0

 

 

0

 

 

0

 

 

6

ORANGE COLOURED URINE

0

0

22

24

CHROMODACRYORRHEA

0

0

0

1

Miscellaneous

ABNORMALGROWTHOFTEETH(cutregulary)

 

1

 

0

 

0

 

0

 

Normal

NOREMARKABLEOBSERVATIONS

 

 

23

 

 

24

 

 

2

 

 

0


  

Table: 2

 

 BODY WEIGHT (Mean values/grams/Females/Pregnancy period)

 

 

 

Dose: (mg/kg/day)

0

20

125

500

 

DAY

 

0

 

MEAN

 

216 d

 

222

 

220

 

222

 

 

S.D.

14

13

12

11

 

 

N

23

24

24

23

DAY

3

MEAN

240 d

244

242

245

 

 

S.D.

15

14

14

12

 

 

N

23

24

24

23

DAY

6

MEAN

259 d

261

260

263

 

 

S.D.

15

15

14

10

 

 

N

23

24

24

23

DAY

9

MEAN

274 d

275

271

259*

 

 

S.D.

19

17

14

20

 

 

N

23

24

24

23

 

DAY 12

 

MEAN

 

296 d

 

 

296

 

294

 

280**

 

S.D.

22

 

18

16

13

 

N

23

 

24

24

23

DAY 15

MEAN

314 d

 

315

311

299*

 

S.D.

24

 

21

17

15

 

N

23

 

24

24

23

DAY 18

MEAN

356 d

 

354

350

339

 

S.D.

30

 

24

20

18

 

N

23

 

24

24

23

DAY 20

MEAN

393 d

 

389

384

372*

 

S.D.

34

 

27

23

20

 

N

23

 

24

24

23

Statistical key:

d=ANOVA+ dunnett+test

* =p<0.05

**=p<0.01

 

 

 


 

  

Table: 3

 

 

 BODY WEIGHT CHANGE (Mean values/grams/Females/Pregnancy period)

 

 

Dose: (mg/kg/day)

0

20

125

500

 

DAYS 0 TO 3

 

MEAN

 

24 d

 

22

 

23

 

22

 

S.D.

4

4

4

6

 

N

23

24

24

23

mean percent change

MEAN%

11.2

10.1

10.3

10.1

 

DAYS 3 TO 6

 

MEAN

 

18 d

 

17

 

17

 

18

 

S.D.

5

6

5

6

 

N

23

24

24

23

mean percent change

MEAN%

7.7

7.1

7.2

7.4

 

DAYS 6 TO 9

 

MEAN

 

15 d

 

13

 

11

 

-4#

 

S.D.

6

6

5

17

 

N

23

24

24

23

mean percent change

MEAN%

5.7

5.1

4.2

-1.6

 

DAYS 9 TO 12

 

MEAN

 

22 d

 

21

 

23

 

21

 

S.D.

6

5

6

14

 

N

23

24

24

23

mean percent change

MEAN%

8.0

7.7

8.5

8.5

 

DAYS 12 TO 15

 

MEAN

 

19 d

 

19

 

17

 

20

 

S.D.

5

5

5

4

 

N

23

24

24

23

mean percent change

MEAN%

6.3

6.5

6.0

7.0

 

DAYS 15 TO 18

 

MEAN

 

42 d

 

39

 

39

 

39

 

S.D.

7

6

6

6

 

N

23

24

24

23

mean percent change

MEAN%

13.2

12.3

12.7

13.1

 

DAYS 18 TO 20

 

MEAN

 

37 d

 

35

 

34

 

33

 

S.D.

7

7

5

5

 

N

23

24

24

23

meanpercentchange                      MEAN%

10.4

9.9

9.6

9.8

DAYS 6TO20                         MEAN

134 d

128

125

109#

 

S.D.

21

16

14

14

 

N

23

24

24

23

mean percent change

MEAN%

51.7

48.9

48.0

41.6

Statistical key:  d=ANOVA+Dunnett-test  #=p<0.001


  

Table: 4

 NET BODY WEIGHT CHANGE (Mean values/grams/Females/Pregnancy period)

 

 

Dose: (mg/kg/day)

0

20

125

500

GRAVIDUTERUS                           MEAN

83.7 d

84.3

83.5

85.1

S.D.

11.1

10.6

11.6

10.1

N

23

24

24

23

CARCASS                                MEAN

309.4 d

305.1

300.7

286.6#

S.D.

27.4

21.3

17.1

14.3

N

23

24

24

23

NET WEIGHT CHANGE FROM DAY 6 MEAN

50.5 d

43.7

41.0*

24.1#

S.D.

14.4

10.6

9.2

10.3

N

23

24

24

23

  

Statisticalkey:   d=ANOVA+Dunnett-test  

* =p<0.05

 #=p<0.001

CARCASS WEIGHT = TERMINAL BODY WEIGHT MINUS UTERINE WEIGHT

NET WEIGHT CHANGE FROM DAY 6 = CARCASS WEIGHT MINUS DAY 6 BODY WEIGHT


 Table 5

FOOD CONSUMPTION (Mean values/grams per day/Females/Pregnancy period)

 

Dose:(mg/kg/day)                  0               20               125          500

DAYS

0TO

3

MEAN

21 d

22

21

21

 

 

 

S.D.

2

2

2

2

 

 

 

N

23

24

24

23

DAYS

3TO

6

MEAN

25 d

25

25

25

 

 

 

S.D.

2

2

2

2

 

 

 

N

23

24

24

23

DAYS

6TO

9

MEAN

26 d

25

24

17#

 

 

 

S.D.

3

2

2

3

 

 

 

N

23

24

24

23

DAYS

9TO

12

MEAN

27 d

26

26

24#

 

 

 

S.D.

3

3

2

2

 

 

 

N

22

24

24

23

 

DAYS12 TO15

 

MEAN

 

28 d

 

27

 

27

 

25**

 

S.D.

4

3

2

2

 

N

23

24

24

23

DAYS15 TO18

MEAN

29 d

28

29

27*

 

S.D.

4

3

2

2

 

N

23

24

24

23

DAYS18 TO20

MEAN

29 d

28

28

25#

 

S.D.

4

3

2

2

 

N

23

24

24

23

Statistical key:

d=ANOVA+Dunnett-test

* =p<0.05

**=p<0.01 #=p<0.001

 

 


 

 Table 6

SUMMARY OF NECROPSY OBSERVATIONS

 

Dose:(mg/kg/day)                   0               20               125          500

FEMALES                          N             24                24                24        24

NO REMARKABLE

OBSERVATIONS                           24                24                24        24


  

Table: 7

 

 Hysterectommy DATA (Summary table)

 

 

Dose:(mg/kg/day)

 

0

 

20

125

500

 

Pregnant FemalesAliveatTerm

 

N

 

 

23

 

24

 

24

 

23

withTotalResorptions

N

 

0

0

0

0

with all Dead Fetuses

N

 

0

0

0

0

with LiveFetuses

N

 

23

24

24

23

 

CorporaLutea                          TOTAL

 

368

 

395

 

385

 

375

No.peranimal                       MEAN

16.0 d

16.5

16.0

16.3

S.D.

1.7

2.2

2.3

2.0

ImplantationSites                          TOTAL

338

351

343

348

No.peranimal                       MEAN

14.7 d

14.6

14.3

15.1

S.D.

1.9

1.7

1.9

1.5

PreimplantationLoss                          TOTAL

30 f

44

42

27

%

8.2

11.1

10.9

7.2

Fetuses                          N

328

342

333

333

No.peranimal                       MEAN

14.3 d

14.3

13.9

14.5

S.D.

1.8

1.8

2.1

2.0

Alive                    %

100.0

100.0

100.0

100.0

Dead                     %

0.0

0.0

0.0

0.0

LiveFetuses                     N

328 f

342

333

333

% of implantation sites

97.0

97.4

97.1

95.7

No.peranimal                         MEAN

14.3 d

14.3

13.9

14.5

S.D.

1.8

1.8

2.1

2.0

DeadFetuses                     N

0 f

0

0

0

% of implantation sites

0.0

0.0

0.0

0.0

No.peranimal                         MEAN

0.0

0.0

0.0

0.0

S.D.

0.0

0.0

0.0

0.0

Statisticalkey:   d=ANOVA+Dunnett-test

f=Fishersexacttest

 

 

 


 

  

 Table: 7 (continued)

 

 HYSTERECTOMY DATA (Summary table)

 

 

Dose: (mg/kg/day)

0

20

125

500

 

Resorptions+Scars

 

N

 

10 f

 

9

 

10

 

15

%ofimplantationsites

 

3.0

2.6

2.9

4.3

No. peranimal

MEAN

0.4 d

0.4

0.4

0.7

 

S.D.

0.8

0.9

0.7

1.1

 

Implant Scars

 

N

 

0 f

 

0

 

0

 

0

%ofimplantationsites

 

0.0

0.0

0.0

0.0

No. peranimal

MEAN

0.0

0.0

0.0

0.0

 

S.D.

0.0

0.0

0.0

0.0

 

Resorptions: early

 

N

 

9 f

 

7

 

10

 

14

%ofimplantationsites

 

2.7

2.0

2.9

4.0

No. peranimal

MEAN

0.4 d

0.3

0.4

0.6

 

S.D.

0.7

0.7

0.7

1.1

Resorptions:late             N              1f        2

 

0

 

1

%ofimplantationsites                    0.3           0.6

0.0

0.3

No.peranimal                       MEAN

0.0 d

0.1

0.0

0.0

S.D.

0.2

0.3

0.0

0.2

PostimplantationLoss                          TOTAL

10 f

9

10

15

% of implantation sites

3.0

2.6

2.9

4.3

No.peranimal                       MEAN

0.4 d

0.4

0.4

0.7

S.D.

0.8

0.9

0.7

1.1

MaleFetuses                                  N

162 f

167

152

170

%

49.4

48.8

45.6

51.1

FemaleFetuses                                  N

166 f

175

181

163

%

50.6

51.2

54.4

48.9

FetalBodyWeight(g)                           MEAN

3.64 d

3.69

3.79

3.72

S.D.

0.21

0.13

0.23

0.34

MaleFetuses                            MEAN

3.73 d

3.79

3.88

3.82

S.D.

0.24

0.16

0.23

0.36

FemaleFetuses                            MEAN

3.55 d

3.60

3.71

3.61

S.D.

0.19

0.16

0.24

0.30

Statisticalkey:   d=ANOVA+Dunnett-test

f=Fishersexacttest

 

 

 


  

Table: 8

 

   SUMMARY OF FETAL EXTERNAL MALFORMATIONS

 

 

Dose:(mg/kg/day)

0

20

125

500

Litters Evaluated

N

23

24

24

23

Fetuses Evaluated

N

328

342

333

333

Live

N

328

342

333

333

Dead

N

0

0

0

0

 

MOUTH, JAW, PALATE

 

LitterIncidence                             N

0

1

0

0

FetalIncidence                             N

0

1

0

0

CLEFT PALATE

FetalIncidence                             N

 

0 f

 

1

 

0

 

0

%

0.0

0.3

0.0

0.0

LitterIncidence                             N

0 f

1

0

0

%

0.0

4.2

0.0

0.0

AffectedFetuses/Litter                            MEAN%

0.0 d

0.3

0.0

0.0

S.D.

0.0

1.3

0.0

0.0

CLEFT LIP

FetalIncidence                             N

 

0 f

 

1

 

0

 

0

%

0.0

0.3

0.0

0.0

LitterIncidence                             N

0 f

1

0

0

%

0.0

4.2

0.0

0.0

AffectedFetuses/Litter                            MEAN%

0.0 d

0.3

0.0

0.0

S.D.

0.0

1.3

0.0

0.0

 

EYES

 

 

 

 

LitterIncidence                             N

0

1

0

0

FetalIncidence                             N

0

1

0

0

OPEN EYE

FetalIncidence                             N

 

0 f

 

1

 

0

 

0

%

0.0

0.3

0.0

0.0

LitterIncidence                             N

0 f

1

0

0

%

0.0

4.2

0.0

0.0

AffectedFetuses/Litter                            MEAN%

0.0 d

0.3

0.0

0.0

S.D.

0.0

1.3

0.0

0.0

Statisticalkey:   d=ANOVA+Dunnett-test

f=Fishers exact test

 

 

 


  

Table: 8 (continued)

 

  SUMMARY OF FETAL EXTERNAL MALFORMATIONS

 

 

Dose:(mg/kg/day)

0

20

125

500

Litters Evaluated

N

23

24

24

23

Fetuses Evaluated

N

328

342

333

333

Live

N

328

342

333

333

Dead

N

0

0

0

0

 

TRUNK

 

Litter Incidence

N

0

0

0

1

Fetal Incidence

N

0

0

0

1

ANAL ATRESIA

Fetal Incidence

 

N

 

0 f

 

0

 

0

 

1

 

%

0.0

0.0

0.0

0.3

Litter Incidence

N

0 f

0

0

1

 

%

0.0

0.0

0.0

4.3

AffectedFetuses/Litter

MEAN%

0.0 d

0.0

0.0

0.3

 

S.D.

0.0

0.0

0.0

1.3

 

TAIL

 

 

 

 

 

Litter Incidence

N

0

0

0

1

Fetal Incidence

N

0

0

0

1

THREAD-LIKE TAIL

Fetal Incidence

 

N

 

0 f

 

0

 

0

 

1

 

%

0.0

0.0

0.0

0.3

Litter Incidence

N

0 f

0

0

1

 

%

0.0

0.0

0.0

4.3

AffectedFetuses/Litter

MEAN%

0.0 d

0.0

0.0

0.3

 

S.D.

0.0

0.0

0.0

1.3

  

CRANIUM

 

Litter Incidence

N

0

2

0

0

Fetal Incidence

N

0

2

0

0

Statistical key:  d=ANOVA+Dunnett-test   f=Fishers exact test


 

 

Table: 8 (continued)

 

   SUMMARY OF FETAL EXTERNAL MALFORMATIONS

  

Dose: (mg/kg/day)

0

20

125

500

LittersEvaluated                N

23

24

24

23

FetusesEvaluated                N

328

342

333

333

Live                      N

328

342

333

333

Dead                      N

0

0

0

0

EXENCEPHALY

FetalIncidence               N

 

0 f

 

2

 

0

 

0

%

0.0

0.6

0.0

0.0

LitterIncidence              N

0 f

2

0

0

%

0.0

8.3

0.0

0.0

AffectedFetuses/Litter                            MEAN%

0.0 d

0.5

0.0

0.0

S.D.

0.0

1.8

0.0

0.0

 

TOTAL FETAL EXTERNAL MALFORMATIONS

 

 

 

 

FetalIncidence               N

0 f

2

0

1

%

0.0

0.6

0.0

0.3

LitterIncidence              N

0 f

2

0

1

%

0.0

8.3

0.0

4.3

AffectedFetuses/Litter                            MEAN%

0.0 d

0.5

0.0

0.3

S.D.

0.0

1.8

0.0

1.3

Statisticalkey:   d=ANOVA+Dunnett-test

f=Fishers exact test

 

 

 


 

 

Table: 9

 

  SUMMARY OF FETAL EXTERNAL VARIATIONS

 

 

Dose:(mg/kg/day)

0

20

125

500

Litters Evaluated

N

23

24

24

23

Fetuses Evaluated

N

328

342

333

333

Live

N

328

342

333

333

Dead

N

0

0

0

0

 

TOTAL FETALEXTERNAL

Fetal Incidence

 

VARIATIONS

N

 

 

0 f

 

 

0

 

 

0

 

 

0

 

%

0.0

0.0

0.0

0.0

LitterIncidence

N

0 f

0

0

0

 

%

0.0

0.0

0.0

0.0

 

AffectedFetuses/Litter

 

MEAN%

 

0.0

 

0.0

 

0.0

 

0.0

 

S.D.

0.0

0.0

0.0

0.0

Statistical key:  f=Fishers exact test


 

Table: 10

 

 

 SUMMARY OF FETAL SOFT TISSUE MALFORMATIONS

 

 

Dose: (mg/kg/day)

0

20

125

500

LittersEvaluated                                  N

23

24

24

23

FetusesEvaluated                                  N

160

165

159

161

 

BRAIN

 

 

 

 

LitterIncidence                               N

0

1

0

0

FetalIncidence                               N

0

1

0

0

HYDROCEPHALY

FetalIncidence                               N

 

0 f

 

1

 

0

 

0

%

0.0

0.6

0.0

0.0

LitterIncidence                               N

0 f

1

0

0

%

0.0

4.2

0.0

0.0

AffectedFetuses/Litter                            MEAN%

0.0 d

0.5

0.0

0.0

S.D.

0.0

2.6

0.0

0.0

 

HEART

 

 

 

 

LitterIncidence                               N

0

2

3

1

FetalIncidence                               N

0

2

3

1

VENTRICULAR SEPTUM DEFECT

FetalIncidence                               N

 

0 f

 

2

 

3

 

1

%

0.0

1.2

1.9

0.6

LitterIncidence                               N

0 f

2

3

1

%

0.0

8.3

12.5

4.3

AffectedFetuses/Litter                            MEAN%

0.0 d

1.2

2.4

0.9

S.D.

0.0

4.0

6.4

4.2

 

LUNGS

 

 

 

 

LitterIncidence                               N

0

1

0

0

FetalIncidence                               N

0

1

0

0

Statisticalkey:   d=ANOVA+Dunnett-test

f=Fishers exact test

 

 

 


  

Table: 10 (continued)

  

 SUMMARY OF FETAL SOFT TISSUE MALFORMATIONS

 

 

Dose: (mg/kg/day)

0

20

125

500

LittersEvaluated                                  N

23

24

24

23

FetusesEvaluated                                  N

160

165

159

161

SMALL LUNG

FetalIncidence                               N

 

0 f

 

1

 

0

 

0

%

0.0

0.6

0.0

0.0

LitterIncidence                               N

0 f

1

0

0

%

0.0

4.2

0.0

0.0

AffectedFetuses/Litter                            MEAN%

0.0 d

0.5

0.0

0.0

S.D.

0.0

2.6

0.0

0.0

 

DIAPHRAGM

 

 

 

 

LitterIncidence                               N

0

1

0

0

FetalIncidence                               N

0

1

0

0

DIAPHRAGMATIC HERNIA

FetalIncidence                               N

 

0 f

 

1

 

0

 

0

%

0.0

0.6

0.0

0.0

LitterIncidence                               N

0 f

1

0

0

%

0.0

4.2

0.0

0.0

AffectedFetuses/Litter                            MEAN%

0.0 d

0.5

0.0

0.0

S.D.

0.0

2.6

0.0

0.0

 

INTESTINES

 

 

 

 

LitterIncidence                               N

0

0

0

1

FetalIncidence                               N

0

0

0

1

ANAL ATRESIA

FetalIncidence                               N

 

0 f

 

0

 

0

 

1

%

0.0

0.0

0.0

0.6

LitterIncidence                               N

0 f

0

0

1

%

0.0

0.0

0.0

4.3

AffectedFetuses/Litter                            MEAN%

0.0 d

0.0

0.0

0.5

S.D.

0.0

0.0

0.0

2.6

Statisticalkey:   d=ANOVA+Dunnett-test

f=Fishers exact test

 

 

 

 

 Table: 10 (continued)

 

  SUMMARY OF FETAL SOFT TISSUE MALFORMATIONS

 

 

Dose: (mg/kg/day)

0

20

125

500

LittersEvaluated                                  N

23

24

24

23

FetusesEvaluated                                  N

160

165

159

161

 

GONADS

 

 

 

 

LitterIncidence                               N

0

1

0

0

FetalIncidence                               N

0

1

0

0

SUPERNUMERARY GONAD

FetalIncidence                               N

 

0 f

 

1

 

0

 

0

%

0.0

0.6

0.0

0.0

LitterIncidence                               N

0 f

1

0

0

%

0.0

4.2

0.0

0.0

AffectedFetuses/Litter                            MEAN%

0.0 d

0.5

0.0

0.0

S.D.

0.0

2.6

0.0

0.0

 

TOTAL FETALSOFTTISSUEMALFORMATIONS

 

 

 

 

FetalIncidence                               N

0 f

4

3

2

%

0.0

2.4

1.9

1.2

LitterIncidence                               N

0 f

4

3

2

%

0.0

16.7

12.5

8.7

AffectedFetuses/Litter                            MEAN%

0.0 d

2.2

2.4

1.4

S.D.

0.0

5.1

6.4

4.8

Statisticalkey:   d=ANOVA+Dunnett-test

f=Fishers exact test

 

 

 


 

 Table: 11

   

 SUMMARY OF FETAL SOFT TISSUE VARIATIONS

 

 

Dose: (mg/kg/day)

0

20

125

500

LittersEvaluated                                  N

23

24

24

23

FetusesEvaluated                                  N

160

165

159

161

 

HEART

 

 

 

 

LitterIncidence                               N

0

2

0

0

FetalIncidence                               N

0

2

0

0

ENLARGED ATRIAL CHAMBER

FetalIncidence                               N

 

0 f

 

2

 

0

 

0

%

0.0

1.2

0.0

0.0

LitterIncidence                               N

0 f

2

0

0

%

0.0

8.3

0.0

0.0

AffectedFetuses/Litter                            MEAN%

0.0 d

1.2

0.0

0.0

S.D.

0.0

4.0

0.0

0.0

 

SPLEEN

 

 

 

 

LitterIncidence                               N

1

0

0

0

FetalIncidence                               N

1

0

0

0

SPLENOMEGALY

FetalIncidence                               N

 

1 f

 

0

 

0

 

0

%

0.6

0.0

0.0

0.0

LitterIncidence                               N

1 f

0

0

0

%

4.3

0.0

0.0

0.0

AffectedFetuses/Litter                            MEAN%

0.6 d

0.0

0.0

0.0

S.D.

3.0

0.0

0.0

0.0

 

STOMACH

 

 

 

 

LitterIncidence                               N

0

1

0

0

FetalIncidence                               N

0

1

0

0

Statisticalkey:   d=ANOVA+Dunnett-test

f=Fishers exact test

 

 

 


 

 Table: 11 (continued)

 

 UMMARY OF FETAL SOFT TISSUE VARIATIONS

 

 

Dose: (mg/kg/day)

0

20

125

500

LittersEvaluated                                  N

23

24

24

23

FetusesEvaluated                                  N

160

165

159

161

ENLARGED STOMACH

FetalIncidence                               N

 

0 f

 

1

 

0

 

0

%

0.0

0.6

0.0

0.0

LitterIncidence                               N

0 f

1

0

0

%

0.0

4.2

0.0

0.0

AffectedFetuses/Litter                            MEAN%

0.0 d

0.5

0.0

0.0

S.D.

0.0

2.6

0.0

0.0

 

KIDNEYS

 

 

 

 

LitterIncidence                               N

4

3

4

3

FetalIncidence                               N

5

4

4

3

DILATED RENAL PELVIS

FetalIncidence                               N

 

5 f

 

4

 

4

 

3

%

3.1

2.4

2.5

1.9

LitterIncidence                               N

4 f

3

4

3

%

17.4

12.5

16.7

13.0

AffectedFetuses/Litter                            MEAN%

3.3 d

2.8

2.4

2.0

S.D.

8.2

8.8

5.5

5.5

 

URETER

 

 

 

 

LitterIncidence                               N

4

1

3

2

FetalIncidence                               N

6

2

3

4

DILATED URETER

FetalIncidence                               N

 

6 f

 

2

 

3

 

4

%

3.8

1.2

1.9

2.5

LitterIncidence                               N

4 f

1

3

2

%

17.4

4.2

12.5

8.7

AffectedFetuses/Litter                            MEAN%

3.5 d

1.7

1.9

2.8

S.D.

8.9

8.2

5.1

9.6

Statisticalkey:   d=ANOVA+Dunnett-test

f=Fishers exact test

 

 

 


 

 Table: 11 (continued)

 

 

 SUMMARY OF FETAL SOFT TISSUE VARIATIONS

 

 

Dose: (mg/kg/day)

0

20

125

500

LittersEvaluated                                  N

23

24

24

23

FetusesEvaluated                                  N

160

165

159

161

 

BLADDER

 

 

 

 

LitterIncidence                               N

0

0

0

2

FetalIncidence                               N

0

0

0

3

DILATED URETHRA

FetalIncidence                               N

 

0 f

 

0

 

0

 

3

%

0.0

0.0

0.0

1.9

LitterIncidence                               N

0 f

0

0

2

%

0.0

0.0

0.0

8.7

AffectedFetuses/Litter                            MEAN%

0.0 d

0.0

0.0

2.0

S.D.

0.0

0.0

0.0

6.5

TOTAL FETALSOFTTISSUEVARIATIONS

FetalIncidence               N

 

10 f

 

6

 

4

 

5

%

6.3

3.6

2.5

3.1

LitterIncidence                               N

7 f

5

4

3

%

30.4

20.8

16.7

13.0

AffectedFetuses/Litter                            MEAN%

6.1 d

4.0

2.4

3.4

S.D.

11.0

9.3

5.5

9.9

Statisticalkey:   d=ANOVA+Dunnett-test

f=Fishersexacttest

 

 

 


  

Table: 12

  SUMMARY OF FETAL SKELETAL MALFORMATIONS

  

Dose:(mg/kg/day)

 

0

20

125

500

Litters Evaluated

N

23

24

24

23

Fetuses Evaluated

N

168

177

174

172

 

THORACIC VERT.

 

 

 

 

 

Litter Incidence

N

0

1

0

0

Fetal Incidence

N

0

1

0

0

 

THORACIC VERTEBRA(E): FUSED ARCH

FetalIncidence                               N

0 f

1

0

0

%

0.0

0.6

0.0

0.0

LitterIncidence                               N

0 f

1

0

0

%

0.0

4.2

0.0

0.0

AffectedFetuses/Litter                            MEAN%

0.0 d

0.5

0.0

0.0

S.D.

0.0

2.6

0.0

0.0

 

LUMBAR VERT.

 

 

 

 

LitterIncidence                               N

0

1

0

0

FetalIncidence                               N

0

1

0

0

SUPERNUMERARY LUMBAR VERTEBRA

FetalIncidence                               N

 

0 f

 

1

 

0

 

0

%

0.0

0.6

0.0

0.0

LitterIncidence                               N

0 f

1

0

0

%

0.0

4.2

0.0

0.0

AffectedFetuses/Litter                            MEAN%

0.0 d

0.5

0.0

0.0

S.D.

0.0

2.6

0.0

0.0

 

RIB

 

 

 

 

LitterIncidence                               N

0

1

0

0

FetalIncidence                               N

0

1

0

0

Statisticalkey:   d=ANOVA+Dunnett-test

f=Fishersexacttest

 

 

 


 Table: 12 (continued)

 

 

 SUMMARY OF FETAL SKELETAL MALFORMATIONS

  

Dose: (mg/kg/day)

0

20

125

500

LittersEvaluated                N

23

24

24

23

FetusesEvaluated                N

168

177

174

172

FUSED RIBS

FetalIncidence               N

 

0 f

 

1

 

0

 

0

%

0.0

0.6

0.0

0.0

LitterIncidence              N

0 f

1

0

0

%

0.0

4.2

0.0

0.0

AffectedFetuses/Litter   MEAN%

0.0 d

0.5

0.0

0.0

S.D.

0.0

2.6

0.0

0.0

 

SPINE

 

 

 

 

LitterIncidence              N

0

1

0

0

FetalIncidence               N

0

1

0

0

 

PRESENCEOF 28PRE-SACRALVERTEBRAE

 

 

 

 

Fetal Incidence

N

0 f

1

0

0

 

%

0.0

0.6

0.0

0.0

Litter Incidence

N

0 f

1

0

0

 

%

0.0

4.2

0.0

0.0

AffectedFetuses/Litter

MEAN%

0.0 d

0.5

0.0

0.0

 

S.D.

0.0

2.6

0.0

0.0

 

TOTAL FETAL SKELETAL MALFORMATIONS

FetalIncidence               N

0 f

1

0

0

%

0.0

0.6

0.0

0.0

LitterIncidence              N

0 f

1

0

0

%

0.0

4.2

0.0

0.0

AffectedFetuses/Litter   MEAN%

0.0 d

0.5

0.0

0.0

S.D.

0.0

2.6

0.0

0.0

Statisticalkey:   d=ANOVA+Dunnett-test

f=Fishersexacttest

 

 

 


 

 Table: 13

  

  SUMMARY OF FETAL SKELETAL VARIATIONS

  

Dose:(mg/kg/day)

 

0

20

125

500

Litters Evaluated

N

23

24

24

23

Fetuses Evaluated

N

168

177

174

172

 

HEAD-SKULL

 

 

 

 

 

Litter Incidence

N

10

7

7

2

Fetal Incidence

N

14

17

8

2

 

INCOMPLETE OSSIFICATION OF INTERPARIETAL

Fetal Incidence

N

12 f

15

6

2

 

%

7.1

8.5

3.4

1.2

Litter Incidence

N

8 f

7

5

2

 

%

34.8

29.2

20.8

8.7

AffectedFetuses/Litter

MEAN%

6.9 d

9.1

4.0

1.3

 

S.D.

12.7

17.6

8.6

4.5

ENLARGED FONTANEL

Fetal Incidence

 

N

 

1 f

 

0

 

0

 

0

 

%

0.6

0.0

0.0

0.0

Litter Incidence

N

1 f

0

0

0

 

%

4.3

0.0

0.0

0.0

AffectedFetuses/Litter

MEAN%

0.6 d

0.0

0.0

0.0

 

S.D.

3.0

0.0

0.0

0.0

 

INCOMPLETE OSSIFICATION OF PARIETAL

FetalIncidence               N

1 f

5

1

0

%

0.6

2.8

0.6

0.0

LitterIncidence              N

1 f

4

1

0

%

4.3

16.7

4.2

0.0

AffectedFetuses/Litter   MEAN%

0.5 d

2.9

0.5

0.0

S.D.

2.3

6.8

2.6

0.0

Statisticalkey:   d=ANOVA+Dunnett-test

f=Fishers exact test

 

 

 


 

 Table: 13 (continued)

  

 

 SUMMARY OF FETAL SKELETAL VARIATIONS

  

Dose:(mg/kg/day)

0

20

125

500

Litters Evaluated

N

23

24

24

23

Fetuses Evaluated

N

168

177

174

172

 

INCOMPLETE OSSIFICATION OF FRONTAL

Fetal Incidence

N

1 f

2

0

0

 

%

0.6

1.1

0.0

0.0

Litter Incidence

N

1 f

2

0

0

 

%

4.3

8.3

0.0

0.0

AffectedFetuses/Litter

MEAN%

0.5 d

1.0

0.0

0.0

 

S.D.

2.6

3.5

0.0

0.0

 

INCOMPLETE OSSIFICATION OF SUPRAOCCIPITAL

Fetal Incidence

N

1 f

1

2

0

 

%

0.6

0.6

1.1

0.0

Litter Incidence

N

1 f

1

2

0

 

%

4.3

4.2

8.3

0.0

AffectedFetuses/Litter

MEAN%

0.6 d

0.5

1.3

0.0

 

S.D.

3.0

2.6

4.4

0.0

 

INCOMPLETE OSSIFICATION OF NASAL

FetalIncidence               N

0 f

1

0

0

%

0.0

0.6

0.0

0.0

LitterIncidence              N

0 f

1

0

0

%

0.0

4.2

0.0

0.0

AffectedFetuses/Litter   MEAN%

0.0 d

0.5

0.0

0.0

S.D.

0.0

2.6

0.0

0.0

UNOSSIFIED SUPRAOCCIPITAL

FetalIncidence               N

 

0 f

 

1

 

0

 

0

%

0.0

0.6

0.0

0.0

LitterIncidence              N

0 f

1

0

0

%

0.0

4.2

0.0

0.0

AffectedFetuses/Litter   MEAN%

0.0 d

0.5

0.0

0.0

S.D.

0.0

2.6

0.0

0.0

Statisticalkey:   d=ANOVA+Dunnett-test

f=Fishers exact test

 

 

 


 Table: 13 (continued)

 

   SUMMARY OF FETAL SKELETAL VARIATIONS

  

Dose: (mg/kg/day)

0

20

125

500

LittersEvaluated                N

23

24

24

23

FetusesEvaluated                N

168

177

174

172

UNOSSIFIED PARIETAL

FetalIncidence               N

 

0 f

 

1

 

0

 

0

%

0.0

0.6

0.0

0.0

LitterIncidence              N

0 f

1

0

0

%

0.0

4.2

0.0

0.0

AffectedFetuses/Litter                            MEAN%

0.0 d

0.5

0.0

0.0

S.D.

0.0

2.6

0.0

0.0

UNOSSIFIED INTERPARIETAL

FetalIncidence               N

 

0 f

 

1

 

0

 

0

%

0.0

0.6

0.0

0.0

LitterIncidence              N

0 f

1

0

0

%

0.0

4.2

0.0

0.0

AffectedFetuses/Litter                            MEAN%

0.0 d

0.5

0.0

0.0

S.D.

0.0

2.6

0.0

0.0

 

HEAD-OTHERS

 

 

 

 

LitterIncidence              N

10

14

10

4

FetalIncidence               N

19

24

18

5

 

INCOMPLETEOSSIFICATIONOFHYOID

 

 

 

 

Fetal Incidence

N

10 f

20

15

5

 

%

6.0

11.3

8.6

2.9

Litter Incidence

N

8 f

12

8

4

 

%

34.8

50.0

33.3

17.4

AffectedFetuses/Litter

MEAN%

5.6 d

11.5

8.0

2.7

 

S.D.

8.5

15.1

13.9

6.5


 

 Table: 13 (continued)

 

SUMMARY OF FETAL SKELETAL VARIATIONS

 

 

Dose: (mg/kg/day)

0

20

125

500

LittersEvaluated                                 N

23

24

24

23

FetusesEvaluated                                 N

168

177

174

172

UNOSSIFIED HYOID

FetalIncidence                              N

 

9 f

 

3

 

3

 

0

%

5.4

1.7

1.7

0.0

LitterIncidence                              N

5 f

2

2

0

%

21.7

8.3

8.3

0.0

AffectedFetuses/Litter                            MEAN%

5.3 d

2.1

1.7

0.0

S.D.

12.6

7.5

6.3

0.0

 

INCOMPLETEOSSIFICATIONOFMAXILLA

 

 

 

 

Fetal Incidence

N

0 f

1

0

0

 

%

0.0

0.6

0.0

0.0

Litter Incidence

N

0 f

1

0

0

 

%

0.0

4.2

0.0

0.0

AffectedFetuses/Litter

MEAN%

0.0 d

0.5

0.0

0.0

 

S.D.

0.0

2.6

0.0

0.0

 

INCOMPLETE OSSIFICATION OF PALATE

Fetal Incidence

N

0 f

1

0

0

 

%

0.0

0.6

0.0

0.0

Litter Incidence

N

0 f

1

0

0

 

%

0.0

4.2

0.0

0.0

AffectedFetuses/Litter

MEAN%

0.0 d

0.5

0.0

0.0

 

S.D.

0.0

2.6

0.0

0.0

 

CERVICAL VERT.

 

 

 

 

 

Litter Incidence

N

0

1

0

0

Fetal Incidence

N

0

1

0

0


 

 Table: 13 (continued)

  

 SUMMARY OF FETAL SKELETAL VARIATIONS

  

Dose:(mg/kg/day)

0

20

125

500

Litters Evaluated

N

23

24

24

23

Fetuses Evaluated

N

168

177

174

172

 

CERVICAL VERTEBRA(E): INCOMPLETE OSSIFICATION OF ARCH

Fetal Incidence

N

0 f

1

0

0

 

%

0.0

0.6

0.0

0.0

Litter Incidence

N

0 f

1

0

0

 

%

0.0

4.2

0.0

0.0

AffectedFetuses/Litter

MEAN%

0.0 d

0.5

0.0

0.0

 

S.D.

0.0

2.6

0.0

0.0

 

THORACIC VERT.

 

 

 

 

 

Litter Incidence

N

12

9

7

5

Fetal Incidence

N

25

12

15

12

 

THORACIC VERTEBRA(E): INCOMPLETE OSSIFICATION OF CENTRUM

Fetal Incidence

N

22 f

10

11

10

 

%

13.1

5.6

6.3

5.8

Litter Incidence

N

10 f

9

5

5

 

%

43.5

37.5

20.8

21.7

AffectedFetuses/Litter

MEAN%

13.2 d

5.9

6.0

5.6

 

S.D.

22.3

8.3

12.7

12.7

 

THORACIC VERTEBRA(E): BIPARTITE OSSIFICATION OF CENTRUM

Fetal Incidence

N

3 f

3

3

2

 

%

1.8

1.7

1.7

1.2

Litter Incidence

N

3 f

2

3

1

 

%

13.0

8.3

12.5

4.3

AffectedFetuses/Litter

MEAN%

1.7 d

1.8

1.9

1.2

 

S.D.

4.5

6.4

5.1

6.0


 

 Table: 13 (continued)

 SUMMARY OF FETAL SKELETAL VARIATIONS

 

 

Dose:(mg/kg/day)

0

20

125

500

Litters Evaluated

N

23

24

24

23

Fetuses Evaluated

N

168

177

174

172

 

THORACIC VERTEBRA(E): UNOSSIFIED CENTRUM

Fetal Incidence

N

2 f

1

1

0

 

%

1.2

0.6

0.6

0.0

Litter Incidence

N

2 f

1

1

0

 

%

8.7

4.2

4.2

0.0

AffectedFetuses/Litter

MEAN%

1.1 d

0.5

0.7

0.0

 

S.D.

3.6

2.6

3.4

0.0

 

THORACIC VERTEBRA(E): INCOMPLETE OSSIFICATION OF HEMICENTRUM

Fetal Incidence

N

0 f

1

0

0

 

%

0.0

0.6

0.0

0.0

Litter Incidence

N

0 f

1

0

0

 

%

0.0

4.2

0.0

0.0

AffectedFetuses/Litter

MEAN%

0.0 d

0.5

0.0

0.0

 

S.D.

0.0

2.6

0.0

0.0

 

LUMBAR VERT.

 

 

 

 

 

Litter Incidence

N

1

0

0

0

Fetal Incidence

N

1

0

0

0

 

LUMBAR VERTEBRA(E): INCOMPLETE OSSIFICATION OF CENTRUM

Fetal Incidence

N

1 f

0

0

0

 

%

0.6

0.0

0.0

0.0

Litter Incidence

N

1 f

0

0

0

 

%

4.3

0.0

0.0

0.0

AffectedFetuses/Litter

MEAN%

0.5 d

0.0

0.0

0.0

 

S.D.

2.6

0.0

0.0

0.0


 

 Table: 13 (continued)

  

  SUMMARY OF FETAL SKELETAL VARIATIONS

 

Dose:(mg/kg/day)

 

0

20

125

500

Litters Evaluated

N

23

24

24

23

Fetuses Evaluated

N

168

177

174

172

 

SACRAL VERT.

 

 

 

 

 

Litter Incidence

N

1

1

0

0

Fetal Incidence

N

1

1

0

0

 

SACRAL VERTEBRA(E): INCOMPLETE OSSIFICATION OF ARCH

Fetal Incidence

N

1 f

0

0

0

 

%

0.6

0.0

0.0

0.0

Litter Incidence

N

1 f

0

0

0

 

%

4.3

0.0

0.0

0.0

AffectedFetuses/Litter

MEAN%

0.6 d

0.0

0.0

0.0

 

S.D.

3.0

0.0

0.0

0.0

 

SACRAL VERTEBRA(E): INCOMPLETE OSSIFICATION OF CENTRUM

Fetal Incidence

N

0 f

1

0

0

 

%

0.0

0.6

0.0

0.0

Litter Incidence

N

0 f

1

0

0

 

%

0.0

4.2

0.0

0.0

AffectedFetuses/Litter

MEAN%

0.0 d

0.5

0.0

0.0

 

S.D.

0.0

2.6

0.0

0.0

 

CAUDAL VERT.

 

 

 

 

 

Litter Incidence

N

3

6

0

1

Fetal Incidence

N

4

7

0

1

 

CAUDAL VERTEBRA(E): UNOSSIFIED CENTRUM

FetalIncidence               N

3 f

3

0

1

%

1.8

1.7

0.0

0.6

LitterIncidence              N

3 f

2

0

1

%

13.0

8.3

0.0

4.3

AffectedFetuses/Litter   MEAN%

1.9 d

1.4

0.0

0.5

S.D.

5.0

5.1

0.0

2.6

Statisticalkey:   d=ANOVA+Dunnett-test

f=Fishers exact test

 

 

 


 

 Table: 13 (continued)

 

 SUMMARY OF FETAL SKELETAL VARIATIONS

 

 

Dose:(mg/kg/day)

0

20

125

500

Litters Evaluated

N

23

24

24

23

Fetuses Evaluated

N

168

177

174

172

 

CAUDAL VERTEBRA(E): INCOMPLETE OSSIFICATION OF CENTRUM

Fetal Incidence

N

2 f

5

0

0

 

%

1.2

2.8

0.0

0.0

Litter Incidence

N

1 f

5

0

0

 

%

4.3

20.8

0.0

0.0

AffectedFetuses/Litter

MEAN%

1.1 d

2.6

0.0

0.0

 

S.D.

5.2

5.3

0.0

0.0

 

STERNEBRA

 

 

 

 

 

Litter Incidence

N

23

24

24

23

Fetal Incidence

N

161

168

161

157

 

INCOMPLETE OSSIFICATION OF 5th STERNEBRA

Fetal Incidence

N

118 f

119

131

127

 

%

70.2

67.2

75.3

73.8

Litter Incidence

N

23 f

24

24

23

 

%

100.0

100.0

100.0

100.0

AffectedFetuses/Litter

MEAN%

70.0 d

67.0

75.3

73.3

 

S.D.

23.0

19.0

22.5

27.2

 

INCOMPLETE OSSIFICATION OF 6th STERNEBRA

FetalIncidence               N

106 f

103

97

114

%

63.1

58.2

55.7

66.3

LitterIncidence              N

23 f

23

22

22

%

100.0

95.8

91.7

95.7

AffectedFetuses/Litter   MEAN%

63.9 d

58.6

55.0

66.4

S.D.

23.7

28.5

29.0

28.6

Statistical key:   d=ANOVA+Dunnett-test

f=Fishers exact test

 

 

 


 

Table: 13 (continued)

  SUMMARY OF FETAL SKELETAL VARIATIONS

 

 

Dose: (mg/kg/day)

0

20

125

500

LittersEvaluated                N

23

24

24

23

FetusesEvaluated                N

168

177

174

172

UNOSSIFIED 5th STERNEBRA

FetalIncidence               N

 

34 f

 

43

 

22

 

21

%

20.2

24.3

12.6

12.2

LitterIncidence              N

15 f

17

12

10

%

65.2

70.8

50.0

43.5

AffectedFetuses/Litter   MEAN%

21.0 d

24.1

12.7

12.1

S.D.

23.5

20.6

17.0

18.2

UNOSSIFIED 6th STERNEBRA

FetalIncidence               N

 

6 f

 

6

 

5

 

7

%

3.6

3.4

2.9

4.1

LitterIncidence              N

2 f

5

2

4

%

8.7

20.8

8.3

17.4

AffectedFetuses/Litter   MEAN%

3.7 d

3.3

3.3

4.1

S.D.

15.1

7.2

11.5

10.5

 

INCOMPLETE OSSIFICATION OF 1st TO 4th STERNEBRA(E)

Fetal Incidence

N

49 f

32

34

59

 

%

29.2

18.1

19.5

34.3

Litter Incidence

N

18 f

16

16

17

 

%

78.3

66.7

66.7

73.9

AffectedFetuses/Litter

MEAN%

28.5 d

17.8

20.1

32.5

 

S.D.

21.5

17.9

21.6

29.2

 

BIPARTITE OSSIFICATION OF STERNEBRA(E)

FetalIncidence               N

0 f

2

0

3

%

0.0

1.1

0.0

1.7

LitterIncidence              N

0 f

2

0

2

%

0.0

8.3

0.0

8.7

AffectedFetuses/Litter   MEAN%

0.0 d

1.0

0.0

1.6

S.D.

0.0

3.5

0.0

5.7

Statisticalkey:   d=ANOVA+Dunnett-test

f=Fishers exact test

 

 

 


  

Table: 13 (continued)

 SUMMARY OF FETAL SKELETAL VARIATIONS

 

 

Dose:(mg/kg/day)

0

20

125

500

Litters Evaluated

N

23

24

24

23

Fetuses Evaluated

N

168

177

174

172

 

UNOSSIFIED 1st TO 4th STERNEBRA(E)

Fetal Incidence

N

1 f

1

0

0

 

%

0.6

0.6

0.0

0.0

Litter Incidence

N

1 f

1

0

0

 

%

4.3

4.2

0.0

0.0

AffectedFetuses/Litter

MEAN%

0.6 d

0.5

0.0

0.0

 

S.D.

3.0

2.6

0.0

0.0

 

RIB

 

 

 

 

 

Litter Incidence

N

5

8

3

6

Fetal Incidence

N

5

12

5

9

SHORT RIB(S)

Fetal Incidence

 

N

 

2 f

 

5

 

2

 

0

 

%

1.2

2.8

1.1

0.0

Litter Incidence

N

2 f

5

2

0

 

%

8.7

20.8

8.3

0.0

AffectedFetuses/Litter

MEAN%

1.3 d

2.6

1.2

0.0

 

S.D.

4.2

5.1

4.0

0.0

 

SHORT SUPERNUMERARY 14th RIB(S)

Fetal Incidence

N

3 f

7

3

9

 

%

1.8

4.0

1.7

5.2

Litter Incidence

N

3 f

3

1

6

 

%

13.0

12.5

4.2

26.1

AffectedFetuses/Litter

MEAN%

2.0 d

4.2

1.6

5.6

 

S.D.

5.2

14.9

7.7

11.9


 

 Table: 13 (continued)

  SUMMARY OF FETAL SKELETAL VARIATIONS

 

 

Dose: (mg/kg/day)

0

20

125

500

LittersEvaluated                                  N

23

24

24

23

FetusesEvaluated                                  N

168

177

174

172

FULL SUPERNUMERARY 14th RIB(S)

FetalIncidence                               N

 

0 f

 

1

 

0

 

0

%

0.0

0.6

0.0

0.0

LitterIncidence                               N

0 f

1

0

0

%

0.0

4.2

0.0

0.0

AffectedFetuses/Litter                            MEAN%

0.0 d

0.5

0.0

0.0

S.D.

0.0

2.6

0.0

0.0

 

INCOMPLETEOSSIFICATIONOFRIB(S)

 

 

 

 

FetalIncidence                               N

0 f

1

0

0

%

0.0

0.6

0.0

0.0

LitterIncidence                               N

0 f

1

0

0

%

0.0

4.2

0.0

0.0

AffectedFetuses/Litter                            MEAN%

0.0 d

0.5

0.0

0.0

S.D.

0.0

2.6

0.0

0.0

 

METACARPAL BONE

 

 

 

 

LitterIncidence                               N

12

5

9

5

FetalIncidence                               N

30

12

13

20

UNOSSIFIED 4th METACARPAL(S)

FetalIncidence                               N

 

30 f

 

12

 

13

 

20

%

17.9

6.8

7.5

11.6

LitterIncidence                               N

12 f

5

9

5

%

52.2

20.8

37.5

21.7

AffectedFetuses/Litter                            MEAN%

18.2 d

7.2

8.7

11.7

S.D.

28.0

19.0

14.7

29.5


 

 Table: 13 (continued)

 

 

 SUMMARY OF FETAL SKELETAL VARIATIONS

 

 

Dose:(mg/kg/day)

0

20

125

500

Litters Evaluated

N

23

24

24

23

Fetuses Evaluated

N

168

177

174

172

 

INCOMPLETE OSSIFICATION OF 1st TO 3rd METACARPALS

Fetal Incidence

N

0 f

2

0

0

 

%

0.0

1.1

0.0

0.0

Litter Incidence

N

0 f

2

0

0

 

%

0.0

8.3

0.0

0.0

AffectedFetuses/Litter

MEAN%

0.0 d

1.0

0.0

0.0

 

S.D.

0.0

3.5

0.0

0.0

 

METATARSAL BONE

 

 

 

 

 

Litter Incidence

N

1

1

0

0

Fetal Incidence

N

1

1

0

0

 

INCOMPLETE OSSIFICATION OF METATARSAL(S)

Fetal Incidence

N

1 f

1

0

0

 

%

0.6

0.6

0.0

0.0

Litter Incidence

N

1 f

1

0

0

 

%

4.3

4.2

0.0

0.0

AffectedFetuses/Litter

MEAN%

0.6 d

0.5

0.0

0.0

 

S.D.

3.0

2.6

0.0

0.0

 

PELVIS

 

 

 

 

 

Litter Incidence

N

2

2

2

1

Fetal Incidence

N

4

2

2

1

 

INCOMPLETE OSSIFICATION OF PUBIS

FetalIncidence               N

4 f

1

2

1

%

2.4

0.6

1.1

0.6

LitterIncidence              N

2 f

1

2

1

%

8.7

4.2

8.3

4.3

AffectedFetuses/Litter   MEAN%

2.5 d

0.6

1.3

0.7

S.D.

9.3

2.9

4.4

3.5

Statisticalkey:   d=ANOVA+Dunnett-test

f=Fishers exact test

 

 

 


 

Table: 13 (continued)

  

  SUMMARY OF FETAL SKELETAL VARIATIONS

  

Dose:(mg/kg/day)

0

20

125

500

Litters Evaluated

N

23

24

24

23

Fetuses Evaluated

N

168

177

174

172

 

INCOMPLETE OSSIFICATION OF ISCHIUM

Fetal Incidence

N

0 f

1

1

0

 

%

0.0

0.6

0.6

0.0

Litter Incidence

N

0 f

1

1

0

 

%

0.0

4.2

4.2

0.0

AffectedFetuses/Litter

MEAN%

0.0 d

0.5

0.7

0.0

 

S.D.

0.0

2.6

3.4

0.0

UNOSSIFIED PUBIS

Fetal Incidence

 

N

 

0 f

 

1

 

1

 

0

 

%

0.0

0.6

0.6

0.0

Litter Incidence

N

0 f

1

1

0

 

%

0.0

4.2

4.2

0.0

AffectedFetuses/Litter

MEAN%

0.0 d

0.5

0.7

0.0

 

S.D.

0.0

2.6

3.4

0.0

 

INCOMPLETE OSSIFICATION OF ILIUM

FetalIncidence               N

0 f

1

0

0

%

0.0

0.6

0.0

0.0

LitterIncidence              N

0 f

1

0

0

%

0.0

4.2

0.0

0.0

AffectedFetuses/Litter   MEAN%

0.0 d

0.5

0.0

0.0

S.D.

0.0

2.6

0.0

0.0

Statisticalkey:   d=ANOVA+Dunnett-test

f=Fishers exact test

 

 

 


 

 

Table: 13 (continued)

   

 SUMMARY OF FETAL SKELETAL VARIATIONS

 

 

Dose: (mg/kg/day)

0

20

125

500

 

TOTAL FETALSKELETAL

Fetal Incidence

 

VARIATIONS

N

 

 

163 f

 

 

171

 

 

164

 

 

160

 

%

97.0

96.6

94.3

93.0

LitterIncidence

N

23 f

24

24

23

 

%

100.0

100.0

100.0

100.0

 

AffectedFetuses/Litter

 

MEAN%

 

97.3 d

 

96.6

 

94.0

 

92.9

 

S.D.

7.9

8.9

12.9

15.7

Statisticalkey:  d=ANOVA+Dunnett-test   f=Fishersexacttest


 

 

 

 

Conclusions:
Administration of Hydroxybenzomorpholine (A025) to pregnant female rats during gestation elicited maternal toxicity at 500 mg/kg/day, consisting of reduced body weight gain and food intake. At 125 mg/kg/day, the maternal conditions were generally unaffected by treatment, except the net body weight change which was slightly but statistically significantly decreased from controls. This slight change was considered to bear no biological significance in the absence of associated changes in body weight gain or carcass weight. No effects were noted in the dams at 20 mg/kg/day.
There were no effects of treatment on litter parameters or fetuses, apart from a slightly high incidence of short 14th rib at 500 mg/kg which was conservatively attributed to treatment with A025. In the absence of 14th full rib or abnormal number of pre-sacral vertebrae, these slight changes are readily reversible (Foulon, 2000) and were not considered to be adverse.
The No Observed Adverse Effect Levels (NOAEL) for this study are considered to be:
. maternal toxicity: 125 mg/kg/day,
. developmental toxicity: 500 mg/kg/day.
Executive summary:

The test solutions were prepared under nitrogen and stored for up to 9 days at + 4 °C and under nitrogen until used. Hydroxybenzomorpholine was suspended in 0.5% aqueous carboxymethylcellulose and given at 5 ml/kg. These dose levels were selected on the basis of the

results from a preliminary study where dose-related maternal toxicity was observed at 400 and 550 mg/kg/day.

Each animal was checked at least twice on dosing days and once on other days for clinical signs and death. Body weight was recorded on GD 0, 3, 6, 9 12, 15 18 and 20. Food intake, was recorded over intervals; GD 0-3, 3-6, 6–9, 9-12, 12–15, 15–18, and 18-20.

Post-mortems of all animals were conducted on GD 20. Adrenals, brain, heart, kidneys, liver, Ovaries and intact uterus removed and examined for corpora lutea, implantation sites, the presence of resorption sites (early and late) and foetuses (live, dead and position). Live foetuses

were weighed sexed and checked for gross malformations. Approximately half of the live foetuses where examined for visceral anomalies, and the remaining foetuses were examined for skeletal anomalies.

Results

There were no deaths. Clinical signs were limited to orange-coloured urine in all females at 500 mg/kg/day and most females at 125 mg/kg/day. Presence of coloured urine was considered to be evidence of systemic exposure following oral administration of hydroxybenzomorpholine.

Mean body weight gain was statistically significantly decreased at 500 mg/kg/day (-19%) compared with controls, mainly due to a transient body weight loss at the onset of dosing.

Consequently, mean body weight was lower than for controls over the entire dosing period, and carcass weight (body weight minus gravid uterine weight) was also lower at 500 mg/kg/day.

Group mean food intake at 500 mg/kg/day was slightly but statistically significantly decreased in comparison with controls over the entire dosing period.

Litter parameters including group mean foetal weight were similar in all groups. The incidences of foetuses with a ventricular septal defect, (2/165, 3/159 and 1/161 at 20, 125 and 500 mg/kg, respectively), were not considered to be treatment-related in the absence of any dose-response.

Higher foetal and litter incidences of short supplementary ribs were observed at 500 mg/kg/day.

In comparison with controls, these incidences were not very high, but fell outside the laboratory historical control range

Effect on developmental toxicity: via oral route
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
NOAEL
500 mg/kg bw/day
Study duration:
subacute
Species:
rat
Quality of whole database:
Reliability 1 (key study guideline and GLP compliant) and 2 (similar to guideline and GLP compliant) studies.
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

The key study is a GLP compliant OECD 414 test performed to evaluate the potential toxic effects of the substance hydroxybenzomorpholine, on the pregnant female and on embryonic and fetal development, following daily oral administration (gavage) to pregnant female rats, from implantation to the day prior to the scheduled hysterectomy. Three groups of 24 mated female Sprague-Dawley rats received the substance by daily oral administration from day 6 until day 19 post-coitum, at the dose-levels of 20, 125 or 500 mg/kg/day. These dose levels were selected on the basis of the results from a preliminary study where dose-related maternal toxicity was observed at 400 and 550 mg/kg/day. One group of 24 females received the vehicle, 0.5% carboxymethylcellulose, only, under the same experimental conditions and acted as a control group.

Clinical signs and mortality were checked daily. Maternal body weight and food consumption were recorded at designated intervals. On day 20 post-coitum, the dams were sacrificed and subjected to a macroscopic examination of the principal thoracic and abdominal organs. The numbers of corpora lutea, implantation sites, early and late resorptions, and live and dead fetuses were recorded for each female. The fetuses were weighed, sexed and examined for external, visceral or skeletal malformations and variations.

Administration of Hydroxybenzomorpholine to pregnant female rats during gestation elicited maternal toxicity at 500 mg/kg/day, consisting of reduced body weight gain and food intake. At 125 mg/kg/day, the maternal conditions were generally unaffected by treatment, except the net body weight change which was slightly but statistically significantly decreased from controls. This slight change was considered to bear no biological significance in the absence of associated changes in body weight gain or carcass weight. No effects were noted in the dams at 20 mg/kg/day.

There were no effects of treatment on litter parameters or fetuses, apart from a slightly high incidence of short 14th rib at 500 mg/kg which was conservatively attributed to treatment with A025. In the absence of 14th full rib or abnormal number of pre-sacral vertebrae, these slight changes are readily reversible and were not considered to be adverse.

The No Observed Adverse Effect Levels (NOAEL) for this key study were considered to be 125 mg/kg/day for maternal toxicity and 500 mg/kg/day for developmental toxicity.

A supporting developmental toxicity study via the oral route is also available. In this GLP compliant study (similar to OECD 414 TG), Hydroxybenzomorpholine (suspended in a hydrogel at 2 grams of polysorbate 80 per 100 mL of water for injection) was administered orally (gavage) to pregnant rat from day 6 to day 15 inclusive of gestation at doses of 5, 50 and 500 mg/kg/day (24 mated females per batch). An equivalent control batch was also formed and received the vehicle. The females were necropsied on day 21 of gestation and the reproductive parameters were recorded. An external macroscopic examination was carried out on all the foetuses. 2/3 of the foetuses per litter were examined at the skeletal level, the other third at the visceral level.

No mortality was noted. From the first administration of the product, the animals of the high dose group presented very quickly hypomotricity associated with a palpebral ptosis. The intensity of clinical signs tended to decrease with repeated treatment. At the end of the study, a significant slowdown of 3.5% in weight growth was observed in females treated with 50 mg/kg/day, associated with a significant slowdown in food consumption (5.5%). At the dose level of 500 mg/kg/day between D6 and D7, there was a significant decrease of 3.2% in body weight gain, then weight growth resumed but remained until the end of the study significantly decreased (from 5.6 to 6.7%) compared to the control group. This was associated with a slowing down of the order of 21.7% and 7.3% between D15 and D21, compared to the control group. The reproduction parameters studied were without variation between the treated and control groups. External and internal macroscopic examinations did not reveal any abnormality related to the administration of the test item. Skeletal examination revealed a slight delay in ossification, observed mainly in the skull bones, 5th sternebra, anterior fontanel and metatarsals but without any difference between the treated and control groups. In the high-dose group, there was an increase in the number of fetuses with a 14th rib either in rough state (13.2%) or fully developed (16.3%). This anomaly, of minor order, was considered not detrimental to the subsequent morphological development of the animal and therefore not teratogenic for the species used.  Under the experimental conditions of this study, it was concluded that Hydroxybenzomorpholine administered at doses of 5, 50, and 500 mg/kg/day was devoid of any embryotoxic and/or teratogenic effects in rats.

Justification for classification or non-classification

Based on the results obtained in the two available repeated dose toxicity studies via the oral route (OECD TG 407 and OECD TG 408) , hydroxybenzomorpholine did not induce any relevant adverse effect on male and female rat reproductive organs or tissues. Hence, the substance hydroxybenzomorpholine does not require classification for fertility toxicity according to the CLP criteria.

Based on the available key developmental toxicity study in the rat (OECD TG 414) via the oral route, the NOAEL for maternal toxicity was 125 mg/kg/day and the NOAEL for developmental toxicity value was found to 500 mg/kg bw (highest dose tested). Hence, the substance hydroxybenzomorpholine does not require classification for developmental toxicity according to the CLP criteria.

Additional information