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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:
2002
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2002
Report Date:
2003

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
other: OECD Guideline 422 (Combined Repeated Dose Toxicity Study with the Reproduction / Developmental Toxicity Screening Test)
Deviations:
no
GLP compliance:
yes
Limit test:
no

Test material

Reference
Name:
Unnamed
Type:
Constituent
Type:
Constituent
Test material form:
solid
Details on test material:
- Lion Corporation Lot No 000203
- Overall purity 97.0% (impurities disodium alpha-sulfopalmitate, sodium methylsufate, palmitic acid)

Test animals

Species:
rat
Strain:
other: Crj: CD(SD)IGS

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
olive oil
Details on exposure:
- Pre-mating exposure period: 14 days (males and females)
Analytical verification of doses or concentrations:
no
Duration of treatment / exposure:
- Males: 47 days
- Females: 42-45 days (from 14 days before mating to Day 4 of lactation)
Frequency of treatment:
Once daily
Doses / concentrationsopen allclose all
Dose / conc.:
0 mg/kg bw/day (nominal)
Dose / conc.:
5 mg/kg bw/day (nominal)
Dose / conc.:
20 mg/kg bw/day (nominal)
Dose / conc.:
80 mg/kg bw/day (nominal)
Dose / conc.:
300 mg/kg bw/day (nominal)
No. of animals per sex per dose:
5 males and 5 females
Control animals:
yes, concurrent vehicle

Examinations

Maternal examinations:
CLINICAL OBSERVATIONS
- General condition was observed once a day.

BODYWEIGHT
- Bodyweights of males were determined on Day 1 (before dosing) and on Days 8, 15, 22, 29, 36, 43 and 49 of treatment.
- Bodyweights of females were determined on Day 1 (before dosing), on Days 1, 8 and 15 of treatment, on Days 0, 7, 14 and 20 of gestation, on Days 0 and 4 of lactation, and on the day of autopsy.

FOOD CONSUMPTION
- Food consumption by males was determined on Days 1, 8, 15, 22, 29, 36, 43 and 48 of treatment.
- Food consumption by females was determined on Days 1, 8 and 15 of treatment, on Days 0, 7, 14 and 20 of gestation, and on Days 0 and 4 of lactation.
- Food consumption of males and females was not determined during the mating period.

URINALYSIS
- Urinalysis was performed for all males on Day 41 or Day 42 of the administration period.

HEAMATOLOGY AND BIOCHEMISTRY
- Investigations were performed at the time of necropsy (after 48 days for males and 5 days after delivery for females).

ORGAN WEIGHTS
- Organ weights from male animals measured at the time of necropsy were brain; heart; liver; kidney; spleen; adrenal; thymus; testis and epididymis.
- Organ weights from female animals measured at the time of necropsy were brain; heart; liver; kidney;spleen; adrenal; thymus.
- Organ weight was determined for 10 males from the control group; 10 males from the 5 mg/kg bw/day group; 9 males from the 20 mg/kg bw/day group; 10 males from the 80 mg/kg bw/day group; 10 males from the 300 mg/kg bw/day group.
- Organ weight was determined for 7 females from the control group; 8 females from the 5 mg/kg bw/day group; 9 females from the 20 mg/kg bw/day group; 10 females from the 80 mg/kg bw/day group; 9 females from the 300 mg/kg bw/day group.

MICROSCOPIC EXAMINATION
- Organs examined in all animals were brain; spinal cord; intestine; liver; kidney; adrenal; spleen; heart; thymus; thyroid; parathyroid; trachea; lung; uterus; ovary; urinary bladder; ischiadic nerve; bone marrow; mesentery lymph node; mandibular lymph node; submandibular gland.
- The sublingual gland from 5 males and 5 females in the 0 and 300 mg/kg bw/day groups was examined.
- The stomach from 5 males and 5 females in the 5, 20 and 80 mg/kg bw/day groups was examined.
Ovaries and uterine content:
- Numbers of corpora lutea and number of implantation sites were considered.
Fetal examinations:
- Pups were examined for external and internal malformations.
Statistics:
STATISTICAL METHODS
- Dunnett's or Scheffe's test for continuous data.
- Chi square test for quantal data.
Indices:
- Number of pairs with successful copulation.
- Number of pregnant females.
- Copulation index (number of pairs with successful copulation / number of pairs mated * 100).
- Fertility index (number of pregnant animals / number of animals with successful copulation * 100).
- Estrous cycle.
- Number of pregnant females with live pups.
- Gestation length.
- Number of corpora lutea.
- Number of implantation sites.
- Number of pups born.
- Number of pups alive on Day 0 of lactation.
- Sex ratio.
- Number of dead pups.
- Gestation index (number of females with live pups / number of pregnant females * 100).
- Implantation index (number of implants / number of corpora lutea * 100).
- Live birth index (number of live pups born / number of pups born * 100)
- Viability index on Day 4 (number of live pups on Day 4 after birth / number of live pups born * 100).

Results and discussion

Results: maternal animals

General toxicity (maternal animals)

Clinical signs:
effects observed, treatment-related
Description (incidence and severity):
- Transitional softening stools were observed in a few males and females in the 80 and 300 mg/kg bw/day groups.
Mortality:
mortality observed, non-treatment-related
Description (incidence):
- No animal deaths related to treatment with test material took place.
Body weight and weight changes:
effects observed, non-treatment-related
Description (incidence and severity):
- There were no statistically significant changes for males or females.
Food consumption and compound intake (if feeding study):
effects observed, non-treatment-related
Description (incidence and severity):
- No statistically significant changes for males or females.
Food efficiency:
not examined
Water consumption and compound intake (if drinking water study):
not examined
Ophthalmological findings:
not examined
Haematological findings:
effects observed, non-treatment-related
Description (incidence and severity):
- No statistically significant changes for males or females.
Clinical biochemistry findings:
effects observed, treatment-related
Description (incidence and severity):
- An increase in GPT levels and a decrease in triglyceride levels was observed in males from the 300 mg/kg bw/day group.
- No statistically significant differences were observed in females.
- Mean GPT was reported as 41 i.u./L (10 animals; 0 mg/kg bw/day; SD 4 i.u./L); 40 i.u./L (10 animals; 5mg/kg bw/day; SD 6 i.u./L); 38 i.u./L (9 animals; 20 mg/kg bw/day; SD 3 i.u./L); 44 i.u./L (10 animals; 80 mg/kg bw/day; SD 6 i.u./L); 53 i.u./L (10 animals; 300 mg/kg bw/day; SD 7 i.u./L; statistically significant p < 0.01).
- Mean triglyceride was reported as 63 mg/dL (10 animals; 0 mg/kg bw/day; SD 28 mg/dL); 43 mg/dL (10 animals; 5 mg/kg bw/day; SD 18 mg/dL); 50 mg/dL (9 animals; 20 mg/kg bw/day; SD 22 mg/dL); 49 mg/dL (10 animals; 80 mg/kg bw/day; SD 18 mg/dL); 29 mg/dL (10 animals; 300 mg/kg bw/day; SD 13 mg/dL; statistically significant p < 0.05).
Urinalysis findings:
effects observed, non-treatment-related
Description (incidence and severity):
- No statistically significant changes.
Behaviour (functional findings):
not examined
Immunological findings:
not examined
Organ weight findings including organ / body weight ratios:
effects observed, non-treatment-related
Description (incidence and severity):
- No statistically significant changes were observed for males or females.
Gross pathological findings:
effects observed, treatment-related
Description (incidence and severity):
- Thickening of the forestomach mucosa was observed in the 80 mg/kg bw/day group (effect seen in 6 out of 10 males and 10 out of 10 females).
- Thickening of the forestomach mucosa was observed in the 300 mg/kg bw/day group (effect seen in 10 out of 10 males and 9 out of 9 females).
Neuropathological findings:
not examined
Histopathological findings: non-neoplastic:
effects observed, treatment-related
Description (incidence and severity):
- Squamous hyperplasia, erosion and lamina propria and/or submucosa edema and inflammatory infiltration were observed in the forestomach of males and females in the 80 and 300 mg/kg bw/day groups (see table, below).
Histopathological findings: neoplastic:
not examined
Other effects:
not examined
Details on results:
- Reproductive and developmental parameters are shown in the table below.

Effect levels (maternal animals)

Key result
Dose descriptor:
NOAEL
Effect level:
> 300 mg/kg bw/day (nominal)
Based on:
test mat.
Basis for effect level:
other:
Remarks on result:
not determinable due to absence of adverse toxic effects

Maternal abnormalities

Key result
Abnormalities:
no effects observed

Results (fetuses)

Fetal body weight changes:
no effects observed
Reduction in number of live offspring:
no effects observed
Changes in sex ratio:
no effects observed
Changes in postnatal survival:
no effects observed
External malformations:
no effects observed
Skeletal malformations:
no effects observed
Visceral malformations:
no effects observed

Effect levels (fetuses)

Key result
Dose descriptor:
NOAEL
Effect level:
300 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other:
Remarks on result:
not determinable due to absence of adverse toxic effects

Fetal abnormalities

Key result
Abnormalities:
no effects observed

Overall developmental toxicity

Key result
Developmental effects observed:
no
Treatment related:
no

Any other information on results incl. tables

Histopathology results

Males

 

 

 

 

 

Dose (mg/kg bw/day)

0

5

20

80

300

Number of animals examined

5

5

5

5

5

Forestomach

 

 

 

 

 

Hyperplasia, squamous (slight to severe)

0/5

0/5

0/5

4/5

5/5 (p < 0.01)

Erosion (slight

to moderate)

0/5

0/5

0/5

0/5

3/5

Edema, lamina

propria/

submucosa

(slight to

moderate)

0/5

0/5

0/5

0/5

4/5 (p < 0.05)

Cellular

infiltration,

inflammatory

cell, lamina

propria/

submucosa

(slight to

moderate)

0/5

0/5

0/5

0/5

4/5 (p < 0.05)

Females

 

 

 

 

 

Dose (mg/kg

bw/day)

0

5

20

80

300

Number of

animals

examined

5

5

5

5

5

Forestomach

 

 

 

 

 

Hyperplasia,

squamous

(slight to

moderate)

0/5

0/5

0/5

4/5 (p < 0.05)

5/5 (p < 0.01)

Erosion

(moderate)

0/5

0/5

0/5

0/5

1/5

Edema, lamina

propria/

submucosa

(slight to

moderate)

0/5

0/5

0/5

2/5

5/5 (p < 0.01)

Cellular

infiltration,

inflammatory

cell, lamina

propria/

submucosa

(slight)

0/5

0/5

0/5

1/5

2/5

Reproductive and developmental parameters

Dose (mg/kg bw/day)

0

5

20

80

300

Estrous cycle (days)

Mean 4.2

Mean 4.0

Mean 4.0

Mean 4.0

Mean 4.0

SD 0.6

SD 0.00

SD 0.0

SD 0.1

SD 0.1

Number of pairs mated

10

10

10

10

10

Number of pairs copulated

10

10

9

10

10

Copulation index (%)

100

100

90

100

100

Number of pregnant females

7

10

9

10

10

Fertility index (%)

70

100

100

100

100

Number of pregnant females with parturition

7

8

9

10

10

Number of pregnant females with live pups

7

8

9

10

10

Gestation length (days)

Mean 22.4

Mean 22.5

Mean 22.4

Mean 22.6

Mean 22.5

SD 0.5

SD 0.5

SD 0.5

SD 0.5

SD 0.5

Number of corpora lutea

Mean 19.7

Mean 18.4

Mean 19.3

Mean 17.5

Mean 18.2

SD 2.8

SD 4.1

SD 3.3

SD 1.9

SD 2.6

Number of implantation sites

Mean 15.6

Mean 13.2

Mean 14.9

Mean 15.9

Mean 15.3

SD 1.6

SD 6.3

SD 2.4

SD 1.5

SD 2.4

Number of pups born

Mean 15.0

Mean 11.9

Mean 13.1

Mean 14.8

Mean 13.9

SD 1.7

SD 6.4

SD 4.0

SD 2.0

SD 1.7

Delivery index (%)

Mean 96.3

Mean 73.8

Mean 85.4

Mean 93.3

Mean 91.4

SD 4.8

SD 39.4

SD 20.0

SD 10.1

SD 6.1

Sex ratio (male/female)

0.78

0.98

0.97

0.85

0.88

Number of pups alive on Day 0 of lactation

Mean 15.0

Mean 14.9

Mean 13.0

Mean 14.4

Mean 12.3

SD 1.7

SD 1.6

SD 3.9

SD 2.0

SD 4.2

Live birth index (%)

Mean 100

Mean 100

Mean 99.3

Mean 97.4

Mean 88.3

SD 0

SD 0

SD 2.1

SD 4.6

SD 27.2

Number of pups alive on Day 4 of lactation

Mean 14.7

Mean 14.9

Mean 12.9

Mean 14.2

Mean 12.0

SD 1.4

SD 1.6

SD 3.8

SD 1.8

SD 4.7

Viability index (%)

Mean 98.3

Mean 100

Mean 99.3

Mean 98.7

Mean 89.3

SD 2.9

SD 0

SD 2.2

SD 2.7

SD 31.5

 

Applicant's summary and conclusion

Conclusions:
The NOAEL for reproduction/developmental toxicity was determined to be 300 mg/kg bw/day in rats.
Executive summary:

In a study involving an analogue substance, a combined repeated dose toxicity study with the reproduction/developmental toxicity screening test [OECD 422] was used to assess the effect of test material administered to Crj:CD (SD) IGS rats (10 animals/sex/dose) by gavage at 0, 5, 20, 80, or 300 mg/kg bw/day. Males were dosed for 47 days from day 14 before mating and females were dosed for 42-45 days from day 14 before mating to day 4 of lactation throughout the mating and pregnancy period.Three females in the control group did not become pregnant due to abnormality of spermatogenesis in paired males. No decrease in fertility index was observed in the groups given this compound. There was no compound-related effect on the estrous cyclicity, copulation index, gestation length, numbers of corpora lutea, or number of implantation sites found in dams. No compound-related effects on the number, sex ratio, body weight, or viability were detected in pups on days 0 and 4 of lactation. No abnormal findings considered to be attributable to administration of the test material were observed in dead pups during lactation and pups at scheduled sacrifice. No external or internal malformations were also noted in pups of any groups. Based on these findings, the NOAEL for reproductive/developmental toxicity was considered to be 300 mg/kg bw/day in rats. Reproduction/developmental parameters, i.e. mating, pregnancy, delivery, lactation, and viability and body weight of pups, were not affected by the test material up to 300 mg/kg bw/day. The NOAEL for reproduction/developmental toxicity was considered to be 300 mg/kg bw/day in rats.