Reaction mass of Potassium sodium 3-{[4-({3-(substitutedamino)-4-[(2-sulfonato-4-{[2-(sulfonatooxy)ethyl]sulfonyl}phenyl)diazenyl]phenyl}amino)-6-chloro-heteromonocyl-2-yl]amino}-2-substituted-5-{[2-(sulfonatooxy)ethyl]sulfonyl}benzenesulfonate (1:3:1) and Potassium sodium 3-[(4-{[3-(subsitutedamino)-4-{[4-(ethenylsulfonyl)-2-sulfonatophenyl]diazenyl}phenyl]amino}-6-chloro-heteromonocy-2-yl)amino]-2-substituted-5-{[2-(sulfonatooxy)ethyl]sulfonyl}benzenesulfonate (3:9:3)

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

Study period:

Testing was conducted between 25th June 2014 and 2nd February 2015

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2015

Report date:

2015

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 421 (Reproduction / Developmental Toxicity Screening Test)

Deviations:

yes

Remarks:

Animal Husbandry. (Please see Principles of method if other tah guidleine part)

Principles of method if other than guideline:

Deviation No.1:
Animal Husbandry
Environment

The Study Plan target values for temperature and relative humidity (RH) were 22 ± 3ºC and 50 ± 20% respectively. While there were no deviations from the target range for temperature, the target range for relative humidity was exceeded for extended periods of time during the study due to adverse weather conditions with the maximum humidity achieved being 79.31% RH.

The results of the study indicate that the animals remained in good health and, while it is accepted that these deviations from the target range for relative humidity were less than ideal, overall it is considered that these deviations had no adverse impact on the scientific purpose of the study.

GLP compliance:

yes (incl. QA statement)

Limit test:

no

Test material

Test material information

Constituent 1

Test material form:

solid: particulate/powder

Remarks:

migrated information: powder

Details on test material:

Identification : FAT 40863/A TE
Harlan Description : Red solid
Sponsor Description : Yellowish to brown powder
Purity : 82.1%
Batch Number : BOP 02-13 (BS-ROE1670)
Date Received : 21 January 2014
Storage Conditions : Room temperature in the dark
Expiry Date : 05 June 2018

Test animals

Species:

rat

Strain:

Wistar

Sex:

male/female

Details on test animals or test system and environmental conditions:

Animals and Animal Husbandry:
A sufficient number of male and female Wistar Han™:RccHan™:WIST strain rats were obtained from Harlan Laboratories U.K. Ltd., Blackthorn, Bicester, Oxon, UK. On receipt the animals were examined for signs of ill-health or injury. The animals were acclimatized for eight days during which time their health status was assessed. A total of ninety six animals (forty eight males and forty eight females) were accepted into the study. At the start of treatment the males weighed 278 to 335g, the females weighed 210 to 234g, and were approximately twelve weeks old.

Initially, all animals were housed in groups of four in solid floor polypropylene cages with stainless steel mesh lids and softwood flake bedding (Datesand Ltd., Cheshire, UK). During the pairing phase, the animals were transferred to polypropylene grid floor cages suspended over trays lined with absorbent paper on a one male: one female basis. Following evidence of successful mating, the males were returned to their original cages. Mated females were housed individually during gestation and lactation in solid floor polypropylene cages with stainless steel mesh lids and softwood flakes.

The animals were allowed free access to food and water. A pelleted diet (Rodent 2018C Teklad Global Certified Diet, Harlan Laboratories UK. Ltd., Oxon, UK.) was used. Mains drinking water was supplied from polycarbonate bottles attached to the cage. Environmental enrichment was provided in the form of wooden chew blocks and cardboard fun tunnels (Dates and Ltd., Cheshire, UK) except for paired animals and mated females during the final week of gestation and lactation. Mated females were also given softwood flakes, as bedding, throughout gestation and lactation. The diet, drinking water, bedding and environmental enrichment were considered not to contain any contaminant at a level that might have affected the purpose or integrity of the study.

The animals were housed in a single air-conditioned room within the Harlan Laboratories Ltd., Shardlow, UK Barrier Maintained Rodent Facility. The rate of air exchange was at least fifteen air changes per hour and the low intensity fluorescent lighting was controlled to give twelve hours continuous light and twelve hours darkness. Environmental conditions were continuously monitored by a computerized system, and print-outs of hourly temperatures and humidities are included in the study records. The Study Plan target ranges for temperature and relative humidity were 22 ± 3 °C and 50 ± 20% respectively. Deviations from these targets were considered not to have affected the purpose or integrity of the study; see deviations from Study Plan.

The animals were randomly allocated to treatment groups using a stratified body weight randomization procedure and the group mean body weights were then determined to ensure similarity between the treatment groups. The cage distribution within the holding rack was also randomized. The animals were uniquely identified within the study by an ear punching system routinely used in these laboratories.

Justification:
The rat was selected for this study as it is a readily available rodent species historically used in safety evaluation studies and is acceptable to appropriate regulatory authorities.

Administration / exposure

Route of administration:

oral: gavage

Type of inhalation exposure (if applicable):

other: Not applicable

Vehicle:

water

Remarks:

distilled water

Details on exposure:

PREPARATION OF DOSING SOLUTIONS:
For the purpose of this study the test item was prepared at the appropriate concentrations as a solution in Distilled water. The stability and homogeneity of the test item formulations were determined by Harlan Laboratories Ltd., Shardlow, UK, Analytical Services. Results showed the formulations to be stable for at least 22 days. Formulations were therefore prepared every two weeks and stored at approximately 4 °C in the dark.

DIET PREPARATION
A pelleted diet (Rodent 2018C Teklad Global Certified Diet, Harlan Laboratories U.K. Ltd., Oxon, UK.) was used throughout the study period.

- Rate of preparation of diet (frequency):
Not applicable

- Mixing appropriate amounts with (Type of food):
Not applicable

- Storage temperature of food:
No data

VEHICLE
- Justification for use and choice of vehicle (if other than water):
Not applicable as distilled water was used as the vehicle

Details on mating procedure:

Animals were paired on a 1 male: 1 female basis within each dose group, for a period of up to fourteen days. Cage tray-liners were checked each morning for the presence of ejected copulation plugs and each female was examined for the presence of a copulation plug in the vagina. A vaginal smear was prepared for each female and the stage of estrus or the presence of sperm was recorded. The presence of sperm within the vaginal smear and/or vaginal plug in situ was taken as positive evidence of mating (Day 0 of gestation) and the males were subsequently returned to their original holding cages. Mated females were housed individually during the period of gestation and lactation.

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

The test item concentration of the test samples was determined by HPLC with UV detection.

Instrumental setup:
HPLC: Agilent Technologies 1100 or 1200, incorporating autosampler and workstation
Column: Gemini 5µ C18 (100 x 4.6)
Mobile phase: Eluent A = water
Eluent B = Acetonitrile
Eluent C: 10g tetrabutylammonium bromide in 500 mL of acetonitrile
Time %A %B %C
0 65 30 5
5 0 95 5
7 0 95 5
7.1 65 30 5
12 65 30 5
Flow rate: 1mL/min
UV detector wave length: 254nm
Injection volume: 100 µL
Retention time: 4 to 8 minutes

The formulations investigated during the study were found to comprise the test item in the range of 98 to 105%.

Discussion: The detection system was found to have acceptable linearity. The analytical procedure had acceptable recoveries of the test item in the vehicle. The method of analysis was validated and proven to be suitable for use.

Duration of treatment / exposure:

The test item was administered by gavage to three groups, each of twelve male and twelve female Wistar Han™:RccHan™:WIST strain rats; males were treated for 42 consecutive days and females were treated up to and including Day 4 post partum (including a two week pre-pairing phase, pairing, gestation and early lactation for females), at dose levels of 100, 300 and 1000 mg/kg bw/day.

Frequency of treatment:

The test item was administered daily by gavage using a stainless steel cannula attached to a disposable plastic syringe.

Details on study schedule:

Chronological Sequence of Study:
i. Groups of twelve male and twelve female animals were treated daily at the appropriate dose level throughout the study (except for females during parturition where applicable). The first day of dosing was designated as Day 1 of the study.
ii. On Day 15, animals were paired on a 1 male: 1 female basis within each dose group for a maximum of fourteen days.
iii. Following evidence of mating (designated as Day 0 post coitum) the males were returned to their original cages and females were transferred to individual cages.
iv. Pregnant females were allowed to give birth and maintain their offspring until Day 5 post partum. Litter size, offspring weight and sex, surface righting and clinical signs were also recorded during this period.
v. The male dose groups were killed and examined macroscopically on Day 43.
vi. At Day 5 post partum, all females and surviving offspring were killed and examined macroscopically.

Doses / concentrations

Doses / concentrations 1

Remarks:

Doses / Concentrations:
0 mg/kg/day
Basis:
actual ingested
0 mg/ml

Doses / concentrations 2

Remarks:

Doses / Concentrations:
100 mg/kg/day
Basis:
actual ingested
20 mg/ml

Doses / concentrations 3

Remarks:

Doses / Concentrations:
300 mg/kg/day
Basis:
actual ingested
60 mg/ml

Doses / concentrations 4

Remarks:

Doses / Concentrations:
1000 mg/kg/day
Basis:
actual ingested
200 mg/ml

No. of animals per sex per dose:

12 animals per sex per dose (including control).

Control animals:

yes, concurrent vehicle

Details on study design:

- Dose selection rationale:
The dose levels were chosen based on previous toxicity data.

-Rationale for animal assignment (if not random):
Random

- Other:
The oral route was selected as the most appropriate route of exposure, based on the physical properties of the test material, and the results of the study are beleived to be of value in screening potential adverse effects on reproduction.

Positive control:

Not applicable

Examinations

Parental animals: Observations and examinations:

DETAILED CLINICAL OBSERVATIONS:
Yes
- Time schedule:
All animals were examined for overt signs of toxicity, ill-health and behavioral change immediately before dosing, soon after dosing, and one hour after dosing (except for females during parturition where applicable). All observations were recorded.

BODY WEIGHT:
Yes
- Time schedule for examinations:
Individual body weights were recorded on Day 1 (prior to dosing) and then weekly for males until termination and weekly for females until pairing. During the pairing phase females were weighed daily until mating was confirmed. Body weights were then recorded for females on Days 0, 7, 14 and 20 post coitum, and on Days 1 and 4 post partum.

FOOD CONSUMPTION:
Yes
- During the maturation period, weekly food consumption was recorded for each cage of adults. This was continued for males after the mating phase. During the pre-pairing period, weekly food consumption was recorded for each cage of adults until pairing. This was continued for males after the mating phase. For females showing evidence of mating, food consumption was recorded for the periods covering post coitum Days 0-7, 7-14 and 14-20. For females with live litters, food consumption was recorded during the lactation period (Days 1-4).

FOOD EFFICIENCY:
Yes
- Weekly food efficiency (body weight gain/food intake) was calculated retrospectively for males and for females during the pre-pairing phase. Due to offspring growth and milk production for lactation, food efficiency for females could not be accurately calculated during gestation and lactation.

WATER CONSUMPTION: Yes
- Time schedule for examinations: Water intake was observed daily by visual inspection of water bottles for any overt changes.

PREGNANCY AND PARTURITION
Each pregnant female was observed at approximately 0830, 1230 and 1630 hours and around the period of expected parturition. Observations were carried out at approximately 0830 and 1230 hours at weekends and public holidays. The following was recorded for each female:

i. Date of pairing
ii. Date of mating
iii. Date and time of observed start of parturition
iv. Date and time of observed completion of parturition

Oestrous cyclicity (parental animals):

No Data

Sperm parameters (parental animals):

Parameters examined in [all/P/F1/F2] male parental generations:
The epididymides and testes were removed from terminal kill adult males dissected free from fat and weighed before fixation.

Litter observations:

LITTER DATA:
On completion of parturition (Day 0 post partum), the number of live and dead offspring was recorded. Offspring were individually identified within each litter by tattoo on Day 1 post partum.

For each litter the following was recorded:

i. Number of offspring born
ii. Number of offspring alive recorded daily and reported on Days 1 and 4 post partum
iii. Sex of offspring on Days 1 and 4 post partum
iv. Clinical condition of offspring from birth to Day 5 post partum
v. Individual offspring weights on Days 1 and 4 post partum (litter weights were calculated retrospectively from this data)

Postmortem examinations (parental animals):

POST-MORTEM EXAMINATIONS: Yes
- Sacrifice
Adult males were killed by intravenous overdose of a suitable barbiturate agent followed by exsanguination on Day 43. Adult females were killed by intravenous overdose of sodium pentobarbitone followed by exsanguination on Day 5 post partum. Surviving offspring were terminated via intracardiac overdose of a suitable barbiturate agent. Any females which failed to achieve pregnancy or produce a litter were killed on or after Day 25 post coitum.

- Organs examined:
For all females, the uterus was examined for signs of implantation and the number of uterine implantations in each horn was recorded. This procedure was enhanced; as necessary, by staining the uteri with a 0.5% ammonium polysulphide solution (Salewski 1964). The corpora lutea were also counted. The epididymides and testes were removed from terminal kill adult males dissected free from fat and weighed before fixation.

Histopathology/organ weights:
All adult animals and offspring, including those dying during the study, were subjected to a full external and internal examination, and any macroscopic abnormalities were recorded.

Samples of the following tissues were preserved from all animals from each dose group, in buffered 10% formalin, except where stated:

Coagulating gland Prostate
Epididymides Seminal vesicles
Ovaries Testes
Mammary gland Uterus/Cervix
Pituitary Vagina

Postmortem examinations (offspring):

POST-MORTEM EXAMINATION
Surviving offspring were terminated via intracardiac overdose of a suitable barbiturate agent. All adult animals and offspring, including those dying during the study, were subjected to a full external and internal examination, and any macroscopic abnormalities were recorded.

Statistics:

Statistical Analysis
Where considered appropriate, quantitative data was subjected to statistical analysis to detect the significance of intergroup differences from control; statistical significance was achieved at a level of p<0.05. Statistical analysis was performed on the following parameters:

Body Weight, Body Weight Change, Food Consumption during gestation and lactation, Pre-Coital Interval, Gestation Length, Litter Size, Litter Weight, Sex Ratio, Corpora Lutea, Implantation Sites, Implantation Losses, Viability Indices, Offspring Body Weight, Offspring Body Weight Change, Offspring Surface Righting, Absolute Organ Weights, Body Weight-Relative Organ Weights.

Data were analyzed using the decision tree from the ProvantisTM Tables and Statistics Module .

Data not analyzed by the Provantis data capture system were assessed separately using the the R Environment for Statistical Computing. Initially, the distribution of the data was assessed by the Shapiro-Wilk normality test, followed by assessment of the homogeneity of the data using Bartlett’s test. Where considered appropriate, parametric analysis of the data was applied incorporating analysis of variance (ANOVA), which if significant, was followed by pair-wise comparisons using Dunnett’s test. Where parametric analysis of the data was considered to be unsuitable, non-parametric analysis of the data was performed incorporating the Kruskal-Wallis test which if significant was followed by the Mann-Whitney "U" test. . Dose response relationships were investigated by linear regression.

Due to the preponderance of non-normally distributed data, reproductive parameters (implantation losses, offspring sex ratio and offspring surface righting) were analyzed using non-parametric analyses.

Probability values (p) are presented as follows:
p<0.01 **
p<0.05 *
p>0.05 (not significant)

Reproductive indices:

Reproductive Indices:
Mating Performance and Fertility

The following parameters were calculated from the individual data during the mating period of the parental generation:

i. Pre-coital Interval

Calculated as the time elapsing between initial pairing and the observation of positive evidence of mating.

ii. Fertility Indices

For each group the following were calculated:

Mating Index (%) = Number of animals mated / number of animals paired x 100

Pregnancy Index (%) = Number of pregnant females / number of animals mated x 100

Offspring viability indices:

Litter Responses:
The standard unit of assessment was considered to be the litter, therefore values were first calculated for each litter and the group mean was calculated using their individual litter values. Group mean values included all litters reared to termination (Day 5 of age).
 
       i.           Implantation Losses (%)
 
Group mean percentile pre-implantation and post-implantation loss were calculated for each female/litter as follows:
 
Pre–implantation loss (%) = (Number of corpora lutea - number of implantati on sited / number of corpora lutea ) x 100
 
Post–implantation loss (%) = (Number of implantati on sites - Total number of offspring born) x 100
 
     ii.           Live Birth and Viability Indices
 
The following indices were calculated for each litter as follows:
 
Live Birth Index (%) = Number of offspring alive on Day 1 / Number of offspring born  x 100
 
Viability Index (%) = Number of offspring alive on Day 4 / Number of offspring alive on Day 1  x 100
 
       i.           Sex Ratio (% males)
 
Sex ratio was calculated for each litter value on Days 1 and 4 post partum, using the following formula:
 
 Number of male off spring / total number of off spring x 100

Results and discussion

Results: P0 (first parental generation)

General toxicity (P0)

Clinical signs:

no effects observed

Description (incidence and severity):

Neither the type, incidence or distribution of observed clinical signs indicated an adverse effect of treatment at 100, 300 or 1000 mg/kg bw/day.

Body weight and weight changes:

no effects observed

Description (incidence and severity):

There were no treatment-related effects on body weight gain at 100, 300 or 1000 mg/kg bw/day.

Food consumption and compound intake (if feeding study):

no effects observed

Description (incidence and severity):

There were no treatment-related effects on body weight gain at 100, 300 or 1000 mg/kg bw/day.

Organ weight findings including organ / body weight ratios:

no effects observed

Histopathological findings: non-neoplastic:

no effects observed

Description (incidence and severity):

There were no treatment-related histopathological changes. All of the histopathological findings encountered were considered to have arisen spontaneously or at post mortem.

Other effects:

not specified

Reproductive function / performance (P0)

Reproductive function: oestrous cycle:

not examined

Reproductive function: sperm measures:

not examined

Reproductive performance:

no effects observed

Description (incidence and severity):

There were no treatment-related effects on conception rates for treated animals.

Details on results (P0)

CLINICAL SIGNS AND MORTALITY (PARENTAL ANIMALS):
There were no unscheduled deaths.

Clinical Observations:
Neither the type, incidence or distribution of observed clinical signs indicated an adverse effect of treatment at 100, 300 or 1000 mg/kg bw/day.

Four males treated with 1000 mg/kg bw/day had incidences of orange staining of the fur between Days 1 and 23. Three females treated with 1000 mg/kg bw/day also had incidences of orange stained fur between Days 18 and 21. Similar observations were apparent in one female treated with 300 mg/kg bw/day on Day 16. These observations were considered to reflect the coloration of the test item.

One male treated with 1000 mg/kg bw/day had red/brown staining of the fur between Days 35 and 36. In isolation this observation was considered not to be toxicologically significant.

One female treated with 1000 mg/kg bw/day had increased salivation on Day 21. Observations of this nature are commonly experienced following the oral administration of an unpalatable or slightly irritant test item formulation and in isolation are considered not to be of toxicological importance.


BODY WEIGHT AND FOOD CONSUMPTION (PARENTAL ANIMALS)
There were no treatment-related effects on body weight gain at 100, 300 or 1000 mg/kg bw/day.
There were no treatment-related effects on food consumption or food efficiency at 100, 300 or 1000 mg/kg bw/day.


WATER CONSUMPTION:
Daily visual inspection of water bottles did not indicate any overt differences in water consumption at 100, 300 or 1000 mg/kg bw/day.


REPRODUCTIVE PERFORMANCE:
-Mating:
There were no treatment-related effects on mating performance.

-Fertility:
There were no treatment-related effects on fertility. One female treated with 1000 mg/kg bw/day was non-pregnant. One female treated with 300 mg/kg bw/day had a total litter loss.

-Gestation length:
There were no differences in gestation lengths that indicated an effect of treatment. Gestation lengths were between 22 and 23½ days.


ORGAN WEIGHTS:
There were no toxicologically significant differences in organ weights detected.


GROSS PATHOLOGY (PARENTAL ANIMALS)
Neither the type, incidence or distribution of macroscopic observations indicated an adverse effect of treatment at 100, 300 or 1000 mg/kg bw/day.


HISTOPATHOLOGY (PARENTAL ANIMALS)
There were no treatment-related histopathological changes. All of the histopathological findings encountered were considered to have arisen spontaneously or at post mortem.

Effect levels (P0)

Dose descriptor:

NOEL

Effect level:

1 000 mg/kg bw/day

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: The ‘No Observed Effect Level’ (NOEL) for reproductive/developmental toxicity was considered to be 1000 mg/kg bw/day.

Results: F1 generation

General toxicity (F1)

Clinical signs:

no effects observed

Description (incidence and severity):

There was no sign of clinical signs within the litter to Day 5 of age at the 100, 300 or 1000 mg/kg bw/day dose groups.

Mortality / viability:

no mortality observed

Description (incidence and severity):

There was no effect of treatment on corpora lutea count, pre-implantation loss, numbers of implantations, post-implantation loss, litter size, sex ratio and subsequent offspring survival to Day 5 of age at 100, 300 or 1000 mg/kg bw/day.

Body weight and weight changes:

no effects observed

Description (incidence and severity):

There was no effect of treatment indicated by body weight gain on Day 1 within the 100, 300 or 1000 mg/kg bw/day dose groups.

Sexual maturation:

not examined

Organ weight findings including organ / body weight ratios:

not examined

Gross pathological findings:

no effects observed

Description (incidence and severity):

Neither the type, incidence or distribution of macroscopic observations in the offspring indicated any adverse effect of treatment at 100, 300 or 1000 mg/kg bw/day.

Histopathological findings:

not examined

Details on results (F1)

Litter Responses:
In total, twelve females from control, and 100 mg/kg bw/day dose groups and eleven females from the 300 and 1000 mg/kg bw/day dose groups gave birth to a live litter and successfully reared young to Day 5 of age. The following assessment of litter response was based on all litters reared to termination on Day 5 of lactation/age.

Offspring Litter Size, Sex Ratio and Viability:
There was no effect of treatment on the corpora lutea count, pre-implantation loss, numbers of implantations, post-implantation loss, litter size at birth/Day 1 post partum and subsequent offspring survival to Day 4 post partum at 100, 300 or 1000 mg/kg bw/day. Sex ratio for the offspring was similar in all groups and did not indicate any selective effect of maternal treatment on survival for either sex.

Offspring Growth and Development:
Offspring body weight gain and litter weights at birth and subsequently on Days 1 and 4 post partum were unaffected by maternal treatment at 100, 300 or 1000 mg/kg bw/day. Clinical signs apparent for offspring on the study were typical for the age observed. Neither the incidence or distribution of these observations indicated any adverse effect of maternal treatment on offspring development at 100, 300 or 1000 mg/kg bw/day.

Statistical analysis of surface righting reflex data did not reveal any significant intergroup differences.

NECROPSY:
Offspring
Macroscopic necropsy findings for offspring on the study were typical for the age observed and neither the incidence or distribution of these observations indicated any adverse effect of maternal treatment on offspring development at 100, 300 or 1000 mg/kg bw/day.

Overall reproductive toxicity

Reproductive effects observed:

not specified

Applicant's summary and conclusion

Conclusions:

The oral administration of FAT 40863/A TE to male rats by gavage for a period of 42 consecutive days and to female rats by gavage up to and including Day 4 post partum (including a two week pre-pairing phase, pairing, gestation and early lactation) at dose levels of 100, 300 and 1000 mg/kg bw/day did not result in any toxicologically significant effects of treatment.

The ‘No Observed Effect Level’ (NOEL) for reproductive/developmental toxicity was considered to be 1000 mg/kg bw/day.

Executive summary:

Introduction:

The study was performed to screen for potential adverse effects of the test item on reproduction, including offspring development, and provides an initial hazard assessment for effect on reproduction. The study is compatible with the requirements of the recommendations of the OECD Guidelines for Testing of Chemicals No. 421 “Reproduction/Developmental Toxicity Screening Test” (adopted 27 July 1995).

 

This study was also designed to be compatible with Commission Regulation (EC) No 440/2008 of 30 May 2008 laying down test methods pursuant to Regulation (EC) No 1907/2006 of the European Parliament and of the Council on the Registration, Evaluation, Authorisation and Restriction of Chemicals (REACH).

 

Methods:

The test item was administered by gavage to three groups, each of twelve male and twelve female Wistar Han™:RccHan™:WIST strain rats; males were treated for 42 consecutive days and females were treated up to and including Day 4 post partum (including a two week pre-pairing phase, pairing, gestation and early lactation for females), at dose levels of 100, 300 and 1000 mg/kg bw/day. A control group of twelve males and twelve females was dosed with vehicle alone (Distilled water).

 

Clinical signs, body weight change, dietary intake and water consumption were monitored during the study. 

 

Pairing of animals within each dose group was undertaken on a one male: one female basis within each treatment group on Day 15 of the study, with females subsequently being allowed to litter and rear their offspring to Day 5 of lactation.

 

During the lactation phase, daily clinical observations were performed on all surviving offspring, together with litter size and offspring weights and assessment of surface righting reflex.

 

Adult males were terminated on Day 43, followed by the termination of all females and offspring on Day 5 post partum. Any female which did not produce a pregnancy was terminated on or after Day 25 post coitum. All animals were subjected to a gross necropsy examination and histopathological evaluation of reproductive tissues was performed.

 

 

Results:

Adult Responses

Mortality

There were no unscheduled deaths.

 

Clinical Observations

Neither the type, incidence or distribution of observed clinical signs indicated an adverse effect of treatment at 100, 300 or 1000 mg/kg bw/day.

 

Body Weight

There were no treatment-related effects on body weight gain at 100, 300 or 1000 mg/kg bw/day.

 

Food Consumption

There were no treatment-related effects on food consumption or food efficiency at 100, 300 or 1000 mg/kg bw/day.

 

Water Consumption

Daily visual inspection of water bottles did not indicate any overt differences in water consumption at 100, 300 or 1000 mg/kg bw/day.

 

Reproductive Performance

Mating

There were no treatment-related effects on mating performance.

 

Fertility

There were no treatment-related effects on conception rates for treated animals.

 

Gestation Length

There were no differences in gestation lengths that indicted an effect of treatment.

 

Litter Responses:

Offspring Litter Size, Sex Ratio and Viability:

There was no effect of treatment on corpora lutea count, pre-implantation loss, numbers of implantations, post-implantation loss, litter size, sex ratio and subsequent offspring survival to Day 5 of age at 100, 300 or 1000 mg/kg bw/day.

 

Offspring Growth and Development:

There was no effect of treatment indicated by offspring body weight or body weight gain and litter weights, surface righting ability on Day 1 or clinical signs to Day 5 of age at 100, 300 or 1000 mg/kg bw/day.

 

Pathology:

Necropsy

Neither the type, incidence or distribution of macroscopic observations in adult animals or offspring indicated any adverse effect of treatment at 100, 300 or 1000 mg/kg bw/day.

 

Organ Weights

There were no treatment-related differences in organ weights detected.

 

Histopathology

There were no treatment-related histopathological changes. All of the histopathological findings encountered were considered to have arisen spontaneously or at post mortem.

Conclusion:

The oral administration of FAT 40863/A TE to male rats by gavage for a period of 42 consecutive days and to female rats by gavage up to and including Day 4 post partum (including a two week pre-pairing phase, pairing, gestation and early lactation) at dose levels of 100, 300 and 1000 mg/kg bw/day did not result in any toxicologically significant effects of treatment.

 

The `No Observed Effect Level' (NOEL) for reproductive/developmental toxicity was considered to be 1000 mg/kg bw/day.