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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:
1988-03-08 to 1988-04-15
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
comparable to guideline study with acceptable restrictions

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1988
Report date:
1988

Materials and methods

Test guideline
Qualifier:
equivalent or similar to guideline
Guideline:
OECD Guideline 414 (Prenatal Developmental Toxicity Study)
Deviations:
no
GLP compliance:
yes
Limit test:
no

Test material

Constituent 1
Chemical structure
Reference substance name:
1-[2-(allyloxy)ethyl-2-(2,4-dichlorophenyl)-1H-imidazolium hydrogen sulphate
EC Number:
261-351-5
EC Name:
1-[2-(allyloxy)ethyl-2-(2,4-dichlorophenyl)-1H-imidazolium hydrogen sulphate
Cas Number:
58594-72-2
Molecular formula:
C14H14Cl2N2O.H2O4S
IUPAC Name:
1-[2-(2,4-DICHLOROPHENYL)-2-(PROP-2-EN-1-YLOXY)ETHYL]-1H-IMIDAZOLE SULFATE (1:1)
Test material form:
solid: particulate/powder
Details on test material:
- Product names: FUNGAFLOR® 75C / FUNGAFLOR® 75SP / FUNGAZIL® 750SP / FUNGAFLOR® 75PS / FUNGAZIL® 75SP / FUNGAFLOR® 750SP
- Reference number: JNJ-2634372-ABI / R027180 (product code)
- Physical state: solid
- Appearance: beige solid
Specific details on test material used for the study:
SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: Janssen Pharmaceutica laboratories; lot ZR027180G1A631. Agroform 047701 was used (Imaezalil 498.6 g/L)
- Appearance: off-white to beige coloured powder
- Expiration date of the lot/batch: no data
- Purity test date: 1998-03-04

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: at room temperature in closed containers
- Stability under test conditions: known
- Solubility and stability of the test substance in the solvent/vehicle: no data

FORM AS APPLIED IN THE TEST (if different from that of starting material) : given as a solution or a suspension

Test animals

Species:
rat
Strain:
Sprague-Dawley
Remarks:
OFA.SD. (IOPS Caw)
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: IFFA CREDO , young and healthy virgin female rats
- Age at study initiation: 2-3 months ; sexually mature at the start of the study
- Weight at study initiation: 245.9 - 252.4 g
- Fasting period before study: no
- Housing: plastic cages with 6 females per cage (37x20x15 cm), and isolated as soon as sperm is observed until the end of pregnancy
- Diet (e.g. ad libitum): ad libitum, pelleted rat food, administered in self-raising hoppers
- Water (e.g. ad libitum): ad libitum, administered in drinking bottles
- Acclimation period: at least one week under test conditions

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20 +- 2°C
- Humidity (%): 50 +- 20 %
- Air changes (per hr): 9
- Photoperiod (hrs dark / hrs light): 12h/day

IN-LIFE DATES: no data

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
water
Details on exposure:
PREPARATION OF DOSING SOLUTIONS:
The test article i given orally by gavage as a solution or a suspension and 1mL per 100g body weight is given.

VEHICLE
- Justification for use and choice of vehicle (if other than water): not specified
- Concentration in vehicle: not specified
- Amount of vehicle (if gavage): 1ml/100 g bw
- Lot/batch no. (if required): not specified
- Purity: not specified
Analytical verification of doses or concentrations:
not specified
Details on mating procedure:
- Impregnation procedure: cohoused
- If cohoused:
- M/F ratio per cage: 1F/2M
- Length of cohabitation: until sperm is observed
- No replacement of first male by another male with proven fertility after number of days of unsuccessful pairing
- Further matings after two unsuccessful attempts: no data
- Verification of same strain and source of both sexes: no
- Proof of pregnancy: sperm in vaginal smear referred to as day 1 of pregnancy
- Any other deviations from standard protocol: males are used for breeding and are not dosed
Duration of treatment / exposure:
from day 6 to day 16 of the pregnancy
Frequency of treatment:
daily
Duration of test:
22 days
Doses / concentrationsopen allclose all
Dose / conc.:
40 mg/kg bw/day (nominal)
Dose / conc.:
80 mg/kg bw/day (nominal)
Dose / conc.:
120 mg/kg bw/day (nominal)
Dose / conc.:
0 mg/kg bw/day (nominal)
Remarks:
control group
No. of animals per sex per dose:
24 females per group, 4 groups
Control animals:
yes, concurrent no treatment
Details on study design:
- Dose selection rationale: Dose levels are determined following review of available data in pharmacology, kinetics and toxicity studies. Dose determination aims to include a surely toxic dose and a no-effect dose level.
- Rationale for animal assignment (if not random): random
- Justification for selection of the test system: The rat is selected following review of available data on toxicity, pharmacology and/or pharmacokinetics and also because of the possibility to compare the hereby recorded data with the accumulated control data of rats.
- Route of administration: this route of administration is chosen following review of available data on pharmacology and/or because this route is the intended route for therapeutic applications of the test article in man and/or animal.

Examinations

Maternal examinations:
CAGE SIDE OBSERVATIONS: Yes
- Time schedule: daily
- Cage side observations include examination of signs of waning health, abnormal behaviour or unusual appearance, occurence of untoward clinical effects and manifestations of toxic and pharmacological response, morbidity and mortality.

DETAILED CLINICAL OBSERVATIONS: No

BODY WEIGHT: Yes
- Time schedule for examinations: in the morning of days 1, 6, 17 and 22 of the presumed pregnancy. Body weights are also determined daily during the dosing period (day 6 through day 16) for adjustment of dose.

FOOD CONSUMPTION: Yes
- Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: No
- Individual records are made prior to, during and after dosing or respectively in the morning of day 6 (non-dosing period: day 1 through day 5), in the morning of day 17 (dosing period: day 6 through day 16) and in the morning of day 22 (non dosing period : day 17 through day 21).

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

POST-MORTEM EXAMINATIONS: Yes
- Prior to sacrifice, a complete physical examination is performed on all surviving rats
- Sacrifice on gestation day 22: all surviving females by decapitation
- autopsy is performed as soon as possible after death during the stud or after sacrifice at the end of the study and any macroscopic pathological changes are noted

ORGAN WEIGHT:
- after removing the uterus in toto, the weight of the uterus is recorded.

Ovaries and uterine content:
The ovaries and uterine content was examined after termination: Yes
Examinations included: Gravid uterus weight, Number of corpora lutea, Number of implantations, Number of resorptions, Number of live and dead fetuses, any abnormal condition
- in case a reduced litter size (0 to 3 fetuses) is noted, the Salewski technique is performed in order to discriminate between resorption and pseudopregnancy
Fetal examinations:
- External examinations: Yes
- all per litter
- all live fetuses are individually weighed
- all live and dead fetuses are carefully examined for any external anomalies
- Soft tissue examinations: No
- Skeletal examinations: Radiographic examinations are carried out for all fetuses of all groups. Rat fetuses of each litter are randomized for dissection (half) and if indicated by the results of the radiographic examination for clearing and bone staining with alizarin (half).
- Head examinations: No
- Malformations or fetal anomalies encountered at the moment delivery by caeserean section or at each stage of dissection are noted
Statistics:
Adult rat:
- mortality and pregnancy: Chi-square test
- body weight, bodyweight change and food consumption: Mann-Whitney U test

Litter data:
live fetuses, dead fetuses, resorbed fetuses, litter size, body weight, abnormalities: Mann-Whitney U test
Indices:
no data
Historical control data:
no data

Results and discussion

Results: maternal animals

General toxicity (maternal animals)

Clinical signs:
not specified
Description (incidence and severity):
No data on clinical signs is available
Dermal irritation (if dermal study):
not examined
Mortality:
mortality observed, non-treatment-related
Description (incidence):
One female (n° 34-not pregnant of the 40 mg/kg dosed group) died for unknown reason on day 12. This animal did not show drug- or dose-related effects.
Body weight and weight changes:
effects observed, treatment-related
Description (incidence and severity):
There was an increase in weight among all dams of control as well as dams doses at 40, 80 and 120 mg/kg. Lower body weight was observed at 40, 80 and 120 mg/kg at the end of dosing period (day 17) and at 120 mg/kg at the end of post-dosing perdiod. In addition, body weight change was lower when compared to controls in the 80 and 120 mg/kg.
Food consumption and compound intake (if feeding study):
effects observed, treatment-related
Description (incidence and severity):
Food consumption significantly decreased during dosing in the three dosed groups compared to controls.
Food efficiency:
not examined
Water consumption and compound intake (if drinking water study):
not examined
Ophthalmological findings:
not examined
Haematological findings:
not examined
Clinical biochemistry findings:
not examined
Urinalysis findings:
not examined
Behaviour (functional findings):
not specified
Immunological findings:
not examined
Organ weight findings including organ / body weight ratios:
not specified
Description (incidence and severity):
No further data available
Gross pathological findings:
not specified
Description (incidence and severity):
No further data available
Neuropathological findings:
not examined
Histopathological findings: non-neoplastic:
not examined
Histopathological findings: neoplastic:
not examined
Other effects:
not specified

Maternal developmental toxicity

Number of abortions:
not specified
Pre- and post-implantation loss:
no effects observed
Description (incidence and severity):
No significant difference wasnoticed in the number of implantations between the various groups.
Total litter losses by resorption:
not specified
Early or late resorptions:
effects observed, treatment-related
Description (incidence and severity):
At 80 mg/kg resorption rate showed a tendency for increase (statistically not significant). At 120 mg/kg bw, this increase was statistically significant (p<0.01). At 120 mg/kg this finding resulted in a lower litter size (p<0.01) and a decreased number of live fetuses (p<0.01).
Dead fetuses:
effects observed, treatment-related
Description (incidence and severity):
At 120 mg/kg there was a decreased number of live fetuses (p<0.01)
Changes in pregnancy duration:
not specified
Changes in number of pregnant:
no effects observed
Description (incidence and severity):
Althought somewhat fluctuating between groups, pregnancy rate was comparable between groups:
Control group: 24/24 or 100 %
40 mg/kg: 20/24 or 83%
80 mg/kg: 23/24 or 96%
120 mg/kg: 22/24 or 92%
Other effects:
not specified

Effect levels (maternal animals)

Key result
Dose descriptor:
LOAEL
Effect level:
40 mg/kg bw/day (nominal)
Based on:
test mat.
Basis for effect level:
body weight and weight gain
food consumption and compound intake

Maternal abnormalities

Abnormalities:
no effects observed

Results (fetuses)

Fetal body weight changes:
effects observed, treatment-related
Description (incidence and severity):
Mean weight was comparable between controls and the 40 mg/kg dosed group. At 80 and 120 mg/kg body weight of pups decreased (p<0.05 and p<0.001, respectively).
Reduction in number of live offspring:
effects observed, treatment-related
Description (incidence and severity):
Decreased number of live fetuses observed at 120 mg/kg bw.
Changes in sex ratio:
no effects observed
Description (incidence and severity):
Fetal sex ratios determined for the dosed groups were 0.84, 0.96 and 0.91 respectively whereas in the controls the ratio was 0.76 which is slightly lower value than normally expected.
Changes in litter size and weights:
effects observed, treatment-related
Description (incidence and severity):
Lower litter size at 120 mg/kg bw.
Changes in postnatal survival:
not specified
External malformations:
no effects observed
Skeletal malformations:
effects observed, non-treatment-related
Description (incidence and severity):
Split vertebrae center, rudimentary 14th ribs and wavy ribs are skeletal variations revealed by the X-ray method and confirmed by the alizarin staining. These findings are reguarly encountered in rat fetuses.
Visceral malformations:
effects observed, non-treatment-related
Description (incidence and severity):
Coelosomia and macroglossia are visceral abnormalities that might occur spontaneously and are randomly distributed among control and dosed groups.
Other effects:
not specified
Details on embryotoxic / teratogenic effects:
It is concluded that no teratogenic effects were evidenced in this study.

Effect levels (fetuses)

open allclose all
Key result
Dose descriptor:
LOAEL
Effect level:
80 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
not specified
Basis for effect level:
reduction in number of live offspring
fetal/pup body weight changes
Key result
Dose descriptor:
NOAEL
Effect level:
40 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
not specified
Basis for effect level:
reduction in number of live offspring
fetal/pup body weight changes

Fetal abnormalities

Abnormalities:
no effects observed

Overall developmental toxicity

Key result
Developmental effects observed:
no

Any other information on results incl. tables

Litter data

     

 Fetuses (number expressed as mean+/-SE)     
 dosage groups litter size  N   live dead  resorbed 
 control group  13.9  24  13.8 +/-0.8  0.0  0.4 +/-0.1
 40 mg/kg  12.1  20  12.0 +/-1.1  0.1 +/-0.1  0.5 +/-0.2
 80 mg/kg  13.0  23  13.0 +/-0.9  0.0  2.0 +/-0.7
 120 mg/kg  11.2**  22  11.1 +/-0.9**  0.1 +/-0.1  3.7 +/-0.7***

N: the number of litters examined per group

**p<0.01; ***p<0.001

Body weight

 Dosage groups N  weight of pups (grams) 
 Control  24  5.6 +/-0.1
 40 mg/kg  20  5.5 +/-0.1
 80 mg/kg  22  5.2 +/-0.1*
 120 mg/kg  22  4.6 +/-0.1***

N represents the number of litters examined per group

*p<0.05; ***p<0.001

Applicant's summary and conclusion

Conclusions:
The test substance, when administered orally to pregnant Sprague-Dawley rats from day 6 through day 16 of pregnancy at the dose of 40 mg/kg was slightly maternally toxic (decreased food consumption and body weight at the end of the dosing) but without any adverse effect on litter parameters. Dosing at 80 and 120 mg/kg also resulted in maternal toxicity (decreased food consumption, body weight at the end of dosing and body weight gain) and herwith associated, in slight to moderate embryonal resorption and decreased pup weight. No teratogenic effect was evidenced up to the dose of 120 mg/kg in rats.