2,5-Furandione, reaction products with 2-ethyl-1-hexanamine and triethanolamine

Toxicity to reproduction

Administrative data

Endpoint:

screening for reproductive / developmental toxicity

Remarks:

based on test type (migrated information)

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP Guideline Study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2013

Report date:

2013

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

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

Test material information

Constituent 1

Reference substance name:

(2E)-3-({3-carboxy-3-[(2-ethylhexyl)amino]propanoyl}(2-ethylhexyl)carbamoyl)prop-2-enoic acid; (2E)-3-[(2-ethylhexyl)carbamoyl]prop-2-enoic acid; (2E)-4-{2-[bis(2-hydroxyethyl)amino]ethoxy}-4-oxobut-2-enoic acid; 2-[(2-hydroxyethyl)amino]ethyl (2E)-3-[(2-ethylhexyl)carbamoyl]prop-2-enoate

EC Number:

939-488-3

Cas Number:

1471311-93-9

Molecular formula:

C18 H36 N2 O6

IUPAC Name:

(2E)-3-({3-carboxy-3-[(2-ethylhexyl)amino]propanoyl}(2-ethylhexyl)carbamoyl)prop-2-enoic acid; (2E)-3-[(2-ethylhexyl)carbamoyl]prop-2-enoic acid; (2E)-4-{2-[bis(2-hydroxyethyl)amino]ethoxy}-4-oxobut-2-enoic acid; 2-[(2-hydroxyethyl)amino]ethyl (2E)-3-[(2-ethylhexyl)carbamoyl]prop-2-enoate

Test material form:

liquid

Details on test material:

- Analytical purity: ca. 100 % (UVCB)
- Lot/batch No.: 85979609T0
- Stability under test conditions: The stability of the test item under storage conditions over the study period was guaranteed by the sponsor, and the sponsor holds this responsibility.
- Storage condition of test material: room temperature

Please note: This kind of substance was in the past handle under the CAS nr 85204-21-3.
After analytical characterization the substance was renamed and received a new CAS nr. Today the substance is characterized by the CAS 1471311-93-9 EC Number: 939-488-3 and handled under these Identifiers.

Test animals

Species:

rat

Strain:

Wistar

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River Laboratories
- Age at study initiation: 11-12 weeks (upon receipt)
- Weight at study initiation: the animals were of comparable size and weight
- Housing: individually with the following exceptions:
• During overnight matings, male and female mating partners were housed together in Makrolon type M III cages.
• Pregnant animals and their litters were housed together until PND 4 (end of lactation).
• For motor activity (MA) measurements the animals were housed individually in polycarbonate cages supplied by TECNIPLAST, Hohenpeißenberg, Germany with wire covers from Ehret, Emmendingen, Germany (floor area of about 800 cm2) and small amounts of bedding material
- Diet: ad libitum
- Water: ad libitum
- Acclimation period: at least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20-24°C
- Humidity (%): 30-70%
- Air changes (per hr): 15 air changes per hour

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

other: water containing 5 mg/100 mL Cremophor EL

Details on exposure:

PREPARATION OF DOSING SOLUTIONS:
Korantin MAT was applied as a suspension. To prepare this suspension, the appropriate amount of test substance was weighed out depending on the desired concentration. Then, drinking water containing 5 mg/100 mL Cremophor EL was filled up to the desired volume, subsequently released with a magnetic stirrer. During administration of the test substance, preparations were kept homogeneous by stirring with a magnetic stirrer. The test substance preparations were produced at least once a week and were stored at room temperature. The administration volume was 10 mL/kg body weight.


VEHICLE
- Justification for use and choice of vehicle (if other than water): the vehicle was suitable for this type of test
- Concentration in vehicle: 1, 3 and 10 g/100 mL

Details on mating procedure:

In general, each of the male and female animals was mated overnight in a 1:1 ratio for a maximum of 2 weeks. Throughout the mating period, each female animal was paired with a predetermined male animal from the same test group.

The animals were paired by placing the female in the cage of the male mating partner from about 15.00 h until 07.00 - 09.00 h of the following morning. Deviations from the specified times were possible on weekends and public holidays and were reported in the raw data. A vaginal smear was prepared after each mating and examined for the presence of sperm. If sperm was detected, pairing of the animals was discontinued. The day on which sperm was detected was denoted gestation day (GD) 0 and the following day "GD 1".

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

The analyses of the test-substance preparations were carried out at the Analytical Chemistry Laboratory of Experimental Toxicology and Ecology of BASF SE, Ludwigshafen, Germany.
The stability of the test substance in drinking water containing 5 mg/100 mL Cremophor EL for a period of 7 days at room temperature was proven during the study.
Homogeneity and concentration control analyses of the test-substance preparations were performed in all concentrations at the start of the administration period.
Additionally, samples from all concentrations as reverse samples for concentration control analysis were taken at the end of the study.

Duration of treatment / exposure:

The duration of treatment covered a 2-week pre-mating and mating period in both sexes, approximately 1 week post-mating in males, and the entire gestation period as well as 4 days of lactation in females followed by an additional treatment until one day before sacrifice.

Frequency of treatment:

once daily

Details on study schedule:

N/A

Doses / concentrations

Remarks:

Doses / Concentrations:
0, 100, 300, 1000 mg/kg
Basis:
actual ingested

No. of animals per sex per dose:

10

Control animals:

yes, concurrent vehicle

Details on study design:

- Dose selection rationale: on request of the sponsor
- Rationale for animal assignment (if not random): random, the list of randomization instructions was compiled with a computer.

Positive control:

N/A

Examinations

Parental animals: Observations and examinations:

CAGE SIDE OBSERVATIONS / DETAILED CLINICAL OBSERVATIONS: Yes
A check for moribund and dead animals was made twice daily on working days and once daily on Saturdays, Sundays and public holidays. If animals were in a moribund state, they were sacrificed and necropsied.

A cageside examination was conducted at least once daily for any signs of morbidity, pertinent behavioral changes and signs of overt toxicity. Abnormalities and changes were documented daily for each affected animal.

The littering and lactation behavior of the dams was generally evaluated in the mornings in combination with the daily clinical inspection of the dams. Only particular findings (e.g. inability to deliver) were documented on an individual dam basis.

On weekdays (except public holidays) the parturition behavior of the dams was inspected in the afternoons in addition to the evaluations in the mornings.

The day of littering was considered the 24-hour period from about 15.00 h of one day until about 15.00 h of the following day.

Detailed clinical observations (DCO) were performed in all animals prior to the administration period and thereafter at weekly intervals. The findings were ranked according to the degree of severity, if applicable. The animals were transferred to a standard arena (50 × 37.5 cm with sides of 25 cm high).

The following parameters were examined:

1. abnormal behavior when handled
2. fur
3. skin
4. posture
5. salivation
6. respiration
7. activity/arousal level
8. tremors
9. convulsions
10. abnormal movements
11. impairment of gait
12. lacrimation
13. palpebral closure
14. exophthalmus
15. feces (appearance/consistency)
16. urine
17. pupil size

BODY WEIGHT: Yes
Body weight was determined before the start of the administration period in order to randomize the animals. During the administration period body weight was determined on study day 0 (start of the administration period) and thereafter once a week at the same time of the day (in the morning).
The body weight change of the animals was calculated from these results.

The following exceptions are notable for the female animals:

- During the mating period the parental fe¬males were weighed on the day of positive evidence of sperm (GD 0) and on GD 7, 14 and 20.

- Females with litter were weighed on the day of parturition (PND 0) and on PND 4.

- Females without a litter and without positive evidence of sperm in the vaginal smear were weighed weekly. These body weight data were solely used for the calculations of the dose volume.


FOOD CONSUMPTION: Yes
Generally, food consumption was determined once a week for male and female parental animals, with the following exceptions:

- Food consumption was not determined during the mating period (male and female F0 animals).
- Food consumption of the F0 females with evidence of sperm was determined on GD 0-7, 7-14, 14-20.
- Food consumption of F0 females, which gave birth to a litter, was determined for PND 1-4.
Food consumption was not determined in females without positive evidence of sperm (during the mating period of dams used in parallel) and females without litter (during the lactation period of dams used in parallel).

WATER CONSUMPTION: Yes
Drinking water consumption was monitored by daily visual inspection of the water bottles for any changes in volume.

Oestrous cyclicity (parental animals):

N/A

Sperm parameters (parental animals):

Special attention was given on stages of spermatogenesis in the male gonads upon Histopathology.

Litter observations:

LITTER DATA
Pup number and status at delivery
All pups delivered from the F0 parents (F1 litter) were examined as soon as possible on the day of birth to determine the total number of pups and the number of liveborn and stillborn pups in each litter. At the same time, the pups were also being examined for macroscopically evident changes. Pups, which died before the first determination of their status on the day of birth, were defined as stillborn pups.

Pup viability/mortality
In general, a check was made for any dead or moribund pups twice daily on workdays (once in the morning and once in the afternoon) or as a rule, only in the morning on Saturdays, Sundays or public holidays.

The number and percentage of dead pups on the day of birth (PND 0) and of pups dying between PND 1 - 4 (lactation period) were determined. Pups which died accidentally or were sacrificed due to maternal death were not included in these calculations. The number of live pups/litter was calculated on the day after birth, and on lactation day 4. The viability index was calculated.

Sex ratio
On the day of birth (PND 0) the sex of the pups was determined by observing the distance between the anus and the base of the genital tubercle; normally, the anogenital distance is considerably greater in male than in female pups. The sex of the pups was finally confirmed at necropsy.
The sex ratio was calculated at day 0 and day 4 after birth.

Pup clinical observations
The live pups were examined daily for clinical symptoms (including gross-morphological findings) during the clinical inspection of the dams.

Pup body weight data
The pups were weighed on the day after birth (PND 1) and on PND 4.
Pups' body weight change was calculated from these results.
The individual weights were always determined at about the same time of the day (in the morning).
Pup body weights and pup body weight change are listed for males, females and males + females.
“Runts” were defined on the basis of the body weights on PND 1. "Runts" are pups that weigh less than 75% of the mean weight of the respective control pups.

Postmortem examinations (parental animals):

GROSS PATHOLOGY / HISTOPATHOLOGY: Yes
Necropsy
All parental animals were sacrificed by decapitation under isoflurane anesthesia. The exsanguinated animals were necropsied and assessed by gross pathology.

Organ weights
Weight assessment was carried out on all animals. The following weights were determined:

1. Anesthetized animals
2. Epididymides
3. Testes


The following weights were determined in 5 animals/sex and test group (females with litters, same animals as used for clinical pathology examinations):

1. Adrenal glands
2. Brain
3. Heart
4. Kidneys
5. Liver
6. Spleen
7. Thymus

Organ/ tissue fixation
The following organs / tissues were preserved in neutral-buffered 4% formaldehyde or in modified Davidson’s solution:

1. Adrenal glands
2. All gross lesions
3. Aorta
4. Bone marrow (femur)
5. Brain
6. Cecum
7. Cervix
8. Coagulating glands
9. Colon
10. Duodenum
11. Eyes with optic nerve
12. Esophagus
13. Extraorbital lacrimal gland
14. Epididymides (modified Davidson’s solution)
15. Femur with knee joint
16. Heart
17. Ileum
18. Jejunum (with Peyer’s patches)
19. Kidneys
20. Larynx
21. Liver
22. Lungs
23. Lymph nodes (axillary and mesenteric)
24. Mammary gland (male and female)
25. Nose (nasal cavity)
26. Ovaries (modified Davidson’s solution)
27. Oviducts
28. Pancreas
29. Parathyroid glands
30. Pharynx
31. Pituitary gland
32. Prostate gland
33. Rectum
34. Salivary glands (mandibular and sublingual)
35. Sciatic nerve
36. Seminal vesicles
37. Skeletal muscle
38. Spinal cord (cervical, thoracic and lumbar cord)
39. Spleen
40. Sternum with marrow
41. Stomach (forestomach and glandular stomach)
42. Testes (modified Davidson’s solution)
43. Thymus
44. Thyroid glands
45. Trachea
46. Urinary bladder
47. Uterus
48. Vagina

Paraffin embedding, sectioning and staining
After the organs were fixed, processing, examination by light microscopy and evaluation of findings were performed, staining with Hematoxylin-eosin (H&E):
all gross lesions (all affected animals per group),
control and high dose animals only:
Adrenal glands (5 animals per sex per group, females with litters only, same animals as used for clinical pathology examination)
Bone marrow (femur) (5 animals per sex per group, females with litters only, same animals as used for clinical pathology examination)
Brain (5 animals per sex per group, females with litters only, same animals as used for clinical pathology examination)
Cecum (5 animals per sex per group, females with litters only, same animals as used for clinical pathology examination)
Cervix (all animals per group)
Coagulation glands (all animals per group)
Colon (5 animals per sex per group, females with litters only, same animals as used for clinical pathology examination)
Duodenum (5 animals per sex per group, females with litters only, same animals as used for clinical pathology examination)
Epididymides (all animals per group)
Heart (5 animals per sex per group, females with litters only, same animals as used for clinical pathology examination)
Ileum (5 animals per sex per group, females with litters only, same animals as used for clinical pathology examination)
Jejunum (5 animals per sex per group, females with litters only, same animals as used for clinical pathology examination)
Kidneys (5 animals per sex per group, females with litters only, same animals as used for clinical pathology examination)
Liver (5 animals per sex per group, females with litters only, same animals as used for clinical pathology examination)
Lung (5 animals per sex per group, females with litters only, same animals as used for clinical pathology examination)
Lymph nodes (mesenteric and axillary lymph nodes) (5 animals per sex per group, females with litters only, same animals as used for clinical pathology examination)
Ovaries (all animals per group)
Oviducts (all animals per group)
Peyer’s patches (5 animals per sex per group, females with litters only, same animals as used for clinical pathology examination)
Prostate (all animals per group)
Rectum (5 animals per sex per group, females with litters only, same animals as used for clinical pathology examination)
Sciatic nerve (5 animals per sex per group, females with litters only, same animals as used for clinical pathology examination)
Seminal vesicles (all animals per group)
Spinal cord (cervical, thoracic and lumbar cords) (5 animals per sex per group, females with litters only, same animals as used for clinical pathology examination)
Spleen (5 animals per sex per group, females with litters only, same animals as used for clinical pathology examination)
Stomach (forestomach and glandular stomach) (5 animals per sex per group, females with litters only, same animals as used for clinical pathology examination)
Testes (all animals per group)
Thymus (5 animals per sex per group, females with litters only, same animals as used for clinical pathology examination)
Thyroid glands (5 animals per sex per group, females with litters only, same animals as used for clinical pathology examination)
Trachea (5 animals per sex per group, females with litters only, same animals as used for clinical pathology examination)
Urinary bladder (5 animals per sex per group, females with litters only, same animals as used for clinical pathology examination)
Uterus (all animals per group)
Vagina (all animals per group)

Special attention was given on stages of spermatogenesis in the male gonads.

A correlation between gross lesions and histopathological findings was attempted.

Postmortem examinations (offspring):

Pup necropsy observations
All pups with scheduled sacrifice on PND 4 were sacrificed under isoflurane anesthesia with CO2. All pups were examined externally and eviscerated; their organs were assessed macroscopically.
All stillborn pups and all pups that died before PND 4 were examined externally, eviscerated and their organs were assessed macroscopically.
All pups without notable findings or abnormalities were discarded after their macroscopic evaluation. Animals with notable findings or abnormalities were evaluated on a case-by-case basis, depending on the type of finding noted.

Statistics:

Detailed statistical analyses were conducted

Reproductive indices:

For the males, mating and fertility indices were calculated. For the females, mating, fertility and gestation indices were calculated. The live birth index was calculated for F1 litters and the postimplantation loss (in %) was calculated.

Offspring viability indices:

The viability index and the sex ratio were calculated

Results and discussion

Results: P0 (first parental generation)

Details on results (P0)

CLINICAL SIGNS AND MORTALITY (PARENTAL ANIMALS)
No animal died prematurely in the present study.

All animals of both sexes in test group 3 (1000 mg/kg bw/d) showed salivation within 2 hours after the administration on several days of the study. In test group 2 (300 mg/kg bw/d) male animal Nos. 25, 27 and 30 showed salivation within 2 hours after the administration on several days during the pre-mating period as well on mating day 0 (male animal No. 27, only). Female animal Nos. 122, 123, 126, 128 and 130 of test group 2 (300 mg/kg bw/d) showed salivation within 2 hours after the administration on several days during the pre-mating period. From the temporary, short appearance immediately after dosing it was concluded that salivation was induced by a bad taste of the test substance or local affection of the upper digestive tract.

During the mating phase male animal No. 6 of test group 0 (control group) showed respirations sounds on mating days 5-6. This finding was assessed as being incidental and not related to treatment.

The Detailed Clinical Observations on study days 0, 7, 14, 21, 28 in male animals and on study days 0, 7, 14, 21, 28, 35, 42, 49, 56 in female animals of test groups 0-3 (0, 100, 300 and 1000 mg/kg bw/d) did not reveal any abnormalities.

BODY WEIGHT AND FOOD CONSUMPTION (PARENTAL ANIMALS)
No test substance-related changes in mean body weights or body weight change values were observed in any test group.
On lactation day 4, mean body weights of female animals in test groups 2 and 3 (300 and 1000 mg/kg bw/d) were significantly lower. In addition, mean body weight change value during lactation was significantly lower in female animals of test group 2 (300 mg/kg bw/d) between study days 0-4. However, no significant changes were observed in on the other time points during the lactation period.

In female animals of test group 3 (1000 mg/kg bw/d) food consumption during gestation was significantly lower between study days 7-14 and 14-20. This finding was assumed to be incidental because the mean value was still within the normal range for pregnant animals at the end of gestation. In addition, no significant impacts on body weight data were observed.


REPRODUCTIVE FUNCTION: SPERM MEASURES (PARENTAL ANIMALS)
Stages of spermatogenesis in the male gonads was comparable to controls.

REPRODUCTIVE PERFORMANCE (PARENTAL ANIMALS)
Male reproduction data

For all F0 parental males, which were placed with females to generate F1 pups, mating was confirmed. Thus, the male mating index was 100% in all test groups (0, 100, 300, 1000 mg/kg bw/d).

Fertility was proven for nearly all F0 parental males within the scheduled mating interval to produce F1 litter. Male animal No. 38 of test group 3 (1000 mg/kg bw/d) did not generate pups with female animal No. 138. Thus, the male fertility index ranged between 90% and 100%.

These values reflected the normal range of biological variation inherent in the strain of rats used for this study as all respective values were within the range of the historical control data.

Female reproduction and delivery data
The female mating index calculated after the mating period for F1 litter was 100% for all test groups (0, 100, 300 and 1000 mg/kg bw/d). The mean duration until sperm was detected (GD 0) was 1.6, 2.2, 1.4 and 1.8 days in test groups 0-3 (0, 100, 300 and 1000 mg/kg bw/d, respectively).

All sperm-positive rats delivered pups with the exception of female animal No. 138 (test group 3), which was mated with male animal No. 38 but did not become pregnant. The female fertility index varied between 90% and 100%. Female animal No. 138, which delivered no pups had no implantation sites.
The mean duration of gestation was between 22.0 and 22.2 days and did not show significant differences.

The gestation index reached 100% in all test groups including the control group.

The rate of liveborn pups was not affected by the test substance, as indicated by live birth indices of 99.1% (control group) and 100% (test groups 1, 2 and 3). One single stillborn pup was only seen in control group.

ORGAN WEIGHTS (PARENTAL ANIMALS)
When compared with control group 0 (set to 100%), the mean relative weights of livers (males only) and kidneys (both sexes) were significantly increased in test group 3 (1000 mg/kg bw/d).

The increase in absolute kidney weights of males and females of test group 3 (1000 mg/kg bw/d) was most likely treatment-related due to a dose response-relationship and also altered relative kidney weights. However, as no histopathologic correlate was present the finding was regarded to be not adverse.
The increase of relative liver weights in males of test group 3 (1000 mg/kg bw/d) was also regarded to be most likely treatment-related although no histopathologic finding could explain the weight increase. But as no other parameter with regard to liver cell metabolism was altered it was regarded to be not adverse in nature.

All other mean weight parameters (absolute and relative) did not show significant differences when compared to the control groups

GROSS PATHOLOGY (PARENTAL ANIMALS)
All gross lesions noted were single observations and they were regarded to have developed spontaneously and unrelated to compound and treatment.

Fertility:
The female that was not pregnant and its male mating partner did not show gross lesions in reproduction relevant organs which could explain the lack of offspring.

HISTOPATHOLOGY (PARENTAL ANIMALS)
All findings noted were either single observations or they were biologically equally distributed between control and treatment group. All of them were considered to be incidental or spontaneous in origin and without any relation to treatment.

Fertility:

The investigated not pregnant female as well as its male mating partner of test group 3 (1000 mg/kg bw/d) did not show relevant histopathological findings that could explain infertility.

Effect levels (P0)

Dose descriptor:

NOAEL

Remarks:

systemic, reproductive performance and fertility, development

Effect level:

1 000 mg/kg bw/day (nominal)

Based on:

test mat.

Sex:

male/female

Remarks on result:

other: Generation: P and F1 (migrated information)

Results: F1 generation

Details on results (F1)

LITTER DATA

Pup number and status at delivery
The mean number of delivered pups per dam and the rate of liveborn and stillborn pups reflect the normal range of biological variation inherent in the strain used in this study.

Pup viability/mortality
The viability index as indicator for pup mortality between PND 0 and 4 was 100% for test groups 0 and 1 (control and 100 mg/kg bw/d). In control group one cannibalized pup was found. In test group 2 (300 mg/kg bw/d) the viability index was 99% because of 1 pup found dead and 1 cannibalized pup in different litters. In test group 3 (1000 mg/kg bw/d) the viability index was 98% because of 2 pups found dead in different litters. The remaining pups which belonged to these animals did not show any findings until sacrifice.

The viability index was still within the historical control data and reflected the normal range of biological variation inherent in the strain used in this study.

Sex ratio
The sex distribution and sex ratios of live F1 pups on the day of birth and on PND 4 did not show biologically relevant differences between test groups.

Pup clinical observations
One pup of female animal No. 134 (test group 3, 1000 mg/kg bw/d) showed a thread-like tail on PND 0-3 and a short tail on PND 4.
One pup of control group (female No. 108) was not assessed because it died during interval on PND 0. The surviving F1 pups of any test group did not show clinical signs up to scheduled sacrifice on PND 4.

Pup body weight data
Mean pup body weights/pup body weight changes of all pups in all test groups were comparable to the concurrent control values.
One female runt was seen in test group 0 as well as test group 2 (control group, 300 mg/kg bw/d) on PND 1. One male runt was seen in test group 3 (1000 mg/kg bw/d) on PND1.
All values were within the range of the biological variation inherent in the strain of rats used for this study.

Pup necropsy observations
In 1 pup of test group 3 (1000 mg/kg bw/d; female No. 134) a small tail was observed. This finding was assessed as being spontaneous in nature and without biological relevance as it can also be found in the historical control data. One pup of test group 0 (control group; female No. 107) was not assessed because it was cannibalized on PND4.

Effect levels (F1)

Dose descriptor:

NOAEL

Generation:

F1

Effect level:

1 000 mg/kg bw/day

Based on:

test mat.

Sex:

male/female

Remarks on result:

not determinable due to absence of adverse toxic effects

Overall reproductive toxicity

Reproductive effects observed:

not specified

Any other information on results incl. tables

Korantin MATwas administered orally by gavage to groups of 10 male and 10 female Wistar rats (F0 animals) at dose levels of 0 mg/kg body weight/day (mg/kg bw/d; test group 0), 100 mg/kg bw/d (test group 1), 300 mg/kg bw/d (test group 2)and 1000 mg/kg bw/d (test group 3).

 

Regardingclinical examinations,signs of general systemic toxicity were not observed in male or female parental animals of test groups 1-3 (100, 300 and 1000 mg/kg bw/d) during the entire study period.

Salivation after treatment was seen in all animals of test group 3 (1000 mg/kg bw/d) and sporadically in a few animals of test group 2 (300 mg/kg bw/d). From the temporary, short appearance immediately after dosing it is likely, that this finding was induced by a bad taste of the test substance or local affection of the upper digestive tract. This finding was not considered to be an adverse and toxicologically relevant effect.

 

Fertility indicesfor male and female animals were not impaired by test-substance administration even at a dose level of 1000 mg/kg bw/d. In addition,live birth indicesof pups in all test groups were not influenced.

 

Theviability indexas indicator for pup mortality was not altered dose-dependently.

 

Concerningclinical pathology, no treatment-related, adverse effects were observed up to a dose of the compound of 1000 mg/kg bw/d.

 

Regardingpathology, the treatment of male and female Wistar rats with up to 1000 mg/kg bw/d of the test substance led to treatment-related increase in male and female absolute and relative kidney weights in test group 3 (1000 mg/kg bw/d). In addition, the relative liver weight in males of test group 3 (1000 mg/kg bw/d) was also significantly and treatment-related increased. As there were no histopathologic findings in both organs these weight increases were not regarded to be adverse in nature but an adaptive effect.

No findings on the reproduction tract were observed.

All other findings occurred either individually or were biologically equally distributed over control and treatment groups. They were considered to be incidental or spontaneous in origin and without any relation to treatment.

CONCLUSION

Under the conditions of this Combined Repeated Dose Toxicity Study with the Reproduction/Developmental Toxicity Screening Test the oral administration by gavage of KORANTIN MAT to Wistar rats revealed no signs of systemic toxicity up to a dose level of 1000 mg/kg bw/d in animals of both sexes. Thus, the no observed adverse effect level (NOAEL) for general systemic toxicity was 1000mg/kg bw/d in male and in female Wistar rats.

The NOAEL for reproductive performance and fertility was set to 1000 mg/kg bw/d in male and female Wistar rats.

TheNOAEL for developmental toxicity was effective 1000 mg/kg bw/d.