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Toxicological information

Toxicity to reproduction

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Administrative data

Endpoint:
extended one-generation reproductive toxicity - with F2 generation and developmental neurotoxicity (Cohorts 1A, 1B with extension, 2A and 2B)
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2 April 2021 - 13 July 2022
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2022

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to guideline
Guideline:
other: OECD Guideline 424. Neurotoxicity Study in Rodents
Version / remarks:
21 July 1997
Deviations:
no
Qualifier:
according to guideline
Guideline:
OECD Guideline 443 (Extended One-Generation Reproductive Toxicity Study)
GLP compliance:
yes (incl. QA statement)
Limit test:
no
Justification for study design:
At this time, studies in laboratory animals provide the best available basis for extrapolation to humans and are required to support regulatory submissions. Acceptable models that do not use live animals currently do not exist. The Han Wistar rat was chosen as the animal model for this study as it is an accepted rodent species for nonclinical toxicity testing by regulatory agencies. The total number of animals to be used in this study is considered to be the minimum required to properly characterise the effects of the test item. This study has been designed such that it does not require an unnecessary number of animals to accomplish its objectives.
The oral route of exposure was selected because this is the intended route of human exposure. In the OECD 414 study (prenatal developmental toxicity), female rats dosed at 900 mg/kg bw/day had treatment related clinical signs like piloerection and lethargy, among others, that occurred over several days but mostly resolved by the end of treatment. Females had lower body weights and weight gains from Days 15-20 post coitum and absolute and relative food consumption was also lower Days 6-17 post coitum. There were no unscheduled deaths during the course of the study and no maternal toxicity was observed in the 100 or 300 mg/kg bw/day groups. No developmental toxicity was observed in the 100, 300 and 900 mg/kg bw/day groups. Based on the results in this prenatal developmental toxicity study (OECD 414, conducted with GLP certification) the maternal No Observed Adverse Effect Level (NOAEL) for the test substance was established as being 300 mg/kg bw/day and developmental NOAEL was established as 900 mg/kg bw/day.
The proposed dose levels for this study were: Low dose 100 mg/kg bw/day, Mid dose 300 mg/kg bw/day, High dose 900 mg/kg bw/day on the basis that evidence of systemic
toxicity will be observed at the top dose level.

Test material

Constituent 1
Chemical structure
Reference substance name:
Hydroxycyclohexyl phenyl ketone
EC Number:
213-426-9
EC Name:
Hydroxycyclohexyl phenyl ketone
Cas Number:
947-19-3
Molecular formula:
C13H16O2
IUPAC Name:
1-benzoylcyclohexan-1-ol
Test material form:
solid: particulate/powder
Details on test material:
Purity 99.7-99.9%
Reference
Constituent 1
Chemical structure
Reference substance name:
Hydroxycyclohexyl phenyl ketone
EC Number:
213-426-9
EC Name:
Hydroxycyclohexyl phenyl ketone
Cas Number:
947-19-3
Molecular formula:
C13H16O2
IUPAC Name:
1-benzoylcyclohexan-1-ol
Test material form:
solid: particulate/powder
Details on test material:
Purity 99.7-99.9%
Specific details on test material used for the study:
- Identification: Omnirad 184
- Alternate identification: Hydroxycyclohexyl phenyl ketone
- EC number: 213-426-9
- CAS number: 947-19-3
- Physical description: Light white solid
- Storage condition of test material: Ambient, protected from light
- Stability and homogeneity of the test material in the vehicle/solvent under test conditions (e.g. in the
exposure medium) and during storage: Test Item is stable in the vehicle when prepared and stored
under the same conditions at concentrations bracketing those used in the present study

Test animals

Species:
rat
Strain:
Wistar
Remarks:
Han Wistar (Crl:WI(Han))
Details on species / strain selection:
The Han Wistar rat was chosen as the animal model for this study as it is an accepted rodent species for nonclinical toxicity testing by regulatory agencies, it is also the preferred model for this OECD guideline study.
Sex:
male/female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: Charles River UK, Margate, Kent, UK
- Age at study initiation: ca 6-7 weeks for males and 5-6 weeks for females

- Weight at study initiation: 150 to 250 g (males)/100 to 150 g (females)
- Housing: Group housed (up to 3 or 4 animals of the same sex and same dosing group together). A few days prior to mating, F0 and Cohort 1B males were be transferred to individual cages with solid bottoms. F0 and Cohort 1B females were transferred to these cages for mating. Mated F0 and Cohort 1B females were transferred to individual solid bottomed cages. White paper tissue was supplied as nesting material from Gestation Day 20. F0 and Cohort 1B females with litters were retained in this type of cage until termination. On a suitable day after completion of mating, the F0 and Cohort 1B males were re-housed with their original cage mates.

- Diet (e.g. ad libitum): Special Diet Services VRF1. Ad libitum, except during designated procedures
- Water (e.g. ad libitum): Public supply tap water. Freely available to each animal from water bottles (except during
designated procedures).
- Acclimation period: The F0 animals were allowed to acclimate to the Test Facility rodent toxicology accommodation for 10 days before the commencement of dosing (replacement animals at least 5 days).
ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19 to 23°C
- Humidity (%): 40 - 70%
- Air changes (per hr): Ten or more air changes per hour
- Photoperiod (hrs dark / hrs light): 12 hours light and 12 hours dark (except during designated procedures)
IN-LIFE DATES: From: 12 Apr 2021 To: 21 Dec 2021

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
CMC (carboxymethyl cellulose)
Remarks:
0.5% (w/v) Carboxymethylcellulose (CMC) medium viscosity and 0.1% (w/v) Tween 80 in Milli-Q Water
Details on exposure:
PREPARATION OF DOSING SOLUTIONS:

Dose formulation: Control and Test Item
Frequency of preparation: At least weekly
Storage conditions: Temperature set to maintain 4°C

Dosing formulations were prepared in the vehicle (0.5% (w/v) CMC medium viscosity and 0.1% (w/v) Tween 80 in Milli-Q Water ) based on a method established at the Test Facility at appropriate concentrations to meet dose level requirements.

Formulation of 100 mg/ml stock suspension
• The required amount of test item was weighed into a suitable container.
• 100% of the required amount of vehicle was weighed into the container with the test item.
• The suspension was mixed by magnetic stirring and then high shear mixing until homogenous.
• During high shear mixing the stock formulation was kept within an ice bath in order to keep the formulation as cool as possible.
• The 100 mg/mL stock suspension was diluted to lower concentrations and split into suitable containers for dosing, as required.
The maximum volume of stock suspension prepared was 1000 mL due to mixing issues with preparations early on in the study and for one formulation occasion, multiple stock amounts were prepared as necessary.
High shear mixing was limited to approximately 1-2 hours, but was adjusted if necessary.

Dilution to a lower concentration
• The required amount of 100 mg/ml stock formulation was weighed into a suitable container.
• 100% of the required amount of vehicle was weighed into the container with the 100 mg/ml stock suspension.
• The formulation was mixed by magnetic stirring until homogenous.
The dosing aliquots were removed from the refrigerator and allowed to warm to room temperature for at least 30 minutes before dosing. Aliquots were also stirred for at least 30 minutes before dosing.

VEHICLE
- Justification for use and choice of vehicle: Carboxymethylcellulose, analysis performed in a separate study showed good stability in this vehicle
- Concentration in vehicle: 10, 30, 90mg/mL
- Amount of vehicle: 10ml/kg

Details on mating procedure:
Frequency: F0 - Day 71
Cohort 1B – PND 107-111

Procedure: Pairing was on a 1 male to 1 female basis. During the evening (after 5 pm), females were housed with their allocated co-group male partner. The animals were paired in ascending numerical order. Vaginal lavages were taken early each morning from the day of pairing until mating had occurred and the stage of estrous observed in each vaginal lavage was recorded. The day of detection of a copulatory plug in situ and/or of sperm in the lavage was designated Gestation Day 0.
The pairing period for each pair of animals was a maximum of 14 nights (unless a longer period was deemed appropriate).
If evidence of mating was not observed by the end of the pairing period, the female was separated from the male during the morning following the last night of pairing and treated as if mating had occurred during that night. Procedures for that female were continued as if it had mated on the last night of pairing.
For each female the time taken to show a positive mating sign and the number of failed opportunities to mate (estruses passed without a sign of mating) was evaluated.
Analytical verification of doses or concentrations:
yes
Remarks:
Concentration & Homogeneity, and stability analysis
Details on analytical verification of doses or concentrations:
Analyses was performed by Gas Chromatography (GC) using a validated analytical procedure.

Concentration and Homogeneity Analysis
Sample Allocation: 2 for analysis, 3 for backup for control; 2 for analysis and 3 for backup per stratum for Groups 2-4.
Sampling Containers: Appropriate sized volumetric flasks
Storage Conditions: Temperature set to maintain 4°C, protected from light
Acceptance Criteria: For concentration: mean sample concentration results within or equal to
± 15% of theoretical concentration. Each individual sample concentration result within or equal to ± 20%. For homogeneity, relative standard deviation (RSD) of concentrations of ≤ 10% for each group.

Stability analyses performed previously in conjunction with 426730 demonstrated that the Test Item is stable in the vehicle when prepared and stored under the same conditions at concentrations bracketing those used in the present study.
Duration of treatment / exposure:
F0 Males: 10 weeks prior to mating and continuing throughout and after mating until the day before termination.

F0 Females: 10 weeks prior to mating and continuing throughout mating and gestation until at least LD 21.
Cohort 1A: From PND 21 until the day before necropsy
Surplus Animals: From PND 21 until the day before necropsy
Cohort 1B: From PND 21 until the day before necropsy
Cohort 2A: From PND 21 until the day before necropsy
Frequency of treatment:
Once daily.
From 19 Aug 2021, F1 Group 4 animals were dosed twice daily (approximately 3 hours apart) at 10mL/kg and 450 mg/kg (45 mg/mL), equivalent to 900 mg/kg/day.
From 30 Aug 2021, F1 Group 4 animals were dosed once daily at 10 mL/kg and 900 mg/kg/day.

Details on study schedule:
F0
Animal Arrival (F0): 02 Apr 2021
Replacement Animals: 09 Apr 2021
Initiation of Dosing: 12 Apr 2021
F0 Start of Estrous Assessment: 07 Jun 2021
F0 Pairing for Mating: 21 Jun 2021
F0 Parturition (Expected): from ca 13 Jul 2021
F0 Dams (Lactation Day (LD) 21) and Unselected F1 Pups Necropsies: from ca 03 Aug 2021
F0 Male Necropsies (Including Computer Assisted Sperm Assessment (CASA)): from 26 Aug 2021

F1
First expected Post Natal Day (PND) 1: from 14 Jul 2021
PND 4 Culled Pups Necropsy and Terminal Blood Sampling: from 17 Jul 2021
Initiation of Dosing (Cohorts 1A + surplus animals, 1B and 2A): from ca 04 Aug 2021
PND 21 Unselected Pups Necropsy and Terminal Blood Sampling: from ca 04 Aug 2021
Cohort 2B Necropsies (PND 21): from 04 Aug 2021
Cohort 2A Necropsies (PND 78-80): from 29 Sep 2021
Cohort 1A Necropsies (PND 91) (males include CASA): from 12 Oct 2021
Surplus animals Necropsies (ca PND 52): 06 Sep 2021

F2
Cohort 1B - Start of Estrous Assessment: 19 Oct 2021
Cohort 1B - Mating: from 02 Nov 2021
Cohort 1B – Parturition (Expected): from 24 Nov 2021
Cohort 1B PND4 Culled Pups Necropsy and Terminal Blood Sampling: from 28 Nov 2021
Cohort 1B – Dams (LD 21) And F2 litter Necropsies: from 15 Dec 2021
Cohort 1B Male Necropsies: from 09 Dec 2021
Completion of In-life: 21 Dec 2021 (Last date of necropsy)
Doses / concentrationsopen allclose all
Dose / conc.:
0 mg/kg bw/day (nominal)
Remarks:
Control
Dose / conc.:
100 mg/kg bw/day (nominal)
Dose / conc.:
300 mg/kg bw/day (nominal)
Dose / conc.:
900 mg/kg bw/day (nominal)
No. of animals per sex per dose:
10 animals per sex per dose
Control animals:
yes, concurrent vehicle
Details on study design:
- Dose selection rationale:
- F0 animals were randomly assigned to groups. Males and females were randomised separately.
- Fasting period before blood sampling for clinical biochemistry: animals not fasted
Positive control:
N/A

Examinations

Parental animals: Observations and examinations:
F0 Animals

CAGE SIDE OBSERVATIONS:
Pre and Postdose Observations, All F0 animals.
Frequency: Once predose and up to 4 hours postdose after the end of each group for each sex.

DETAILED CLINICAL OBSERVATIONS:
All F0 animals Weekly; from at least Week 1 and throughout the study.

BODY WEIGHT:
All F0 animals.
Males: weekly beginning Week -1.
Females: weekly beginning Week -1 until pairing for mating and then on GD 0, 7, 14 and 20 and LD 1, 4, 7, 14 and 21.

FOOD CONSUMPTION:
All F0 animals.
Males: weekly from Week -1 until pairing for mating. Then weekly after mating and re-housing.
Females: weekly beginning week -1 until pairing for mating and then on GD 0-7, 7-14 and 14-20, and LD 1-7, 7-14 and 14-21

WATER CONSUMPTION:
All F0 animals. Regular basis throughout the study.

Mortality:
All F0 animals. At least twice daily (once at the start and once towards the end of the working day) beginning upon arrival through termination/release.
Oestrous cyclicity (parental animals):
Estrous Cycle Monitoring (Number of Estrous Cycles and Mean Cycle Length):
All F0 females. From 2 weeks prior to pairing until day of detection of a copulatory plug in situ and/or of sperm in the lavage. Until either mating was detected or the end of the mating period. Also, on the morning of necropsy.
Sperm parameters (parental animals):
Parameters examined in F0 and Cohort 1A Males:

Computer Aided Sperm Assessment (CASA)
From all F0 and Cohort 1A males at terminal euthanasia, the right cauda epididymis was placed in 0.3% BSA in Medium 199 (as per SOP/PAT/069) and the sperm was allowed to “swim out” into the medium. An appropriate dilution of the sperm suspension was prepared and examined using a Hamilton Thorne sperm motility analyser.

Sperm Count and Morphological Analysis
The cauda epididymis was minced and suspended. Dilutions of this sperm suspension were counted using a haemocytometer to obtain a total sperm count which was expressed per cauda epididymis and per gram of cauda epididymis. Optionally, the cauda epididymis was frozen prior to assessment. From all samples of the sperm suspension described in the preceding paragraph, a sperm smear was prepared and stained with eosin Y solution. At least two hundred sperm per animal were evaluated for morphological abnormalities

Spermatid Count
The right testes was decapsulated and homogenised. The number of homogenisation resistant spermatids in dilutions of this suspension were counted using a haemocytometer to obtain a total spermatid count which was expressed per testis and per gram of testis. Optionally, the testis was frozen prior to assessment.

F0 Animals and Cohort 1A
Testis were weighed.
Litter observations:
STANDARDISATION OF LITTERS
For females, on Lactation Day 4, litter size was standardised to 8 pups per litter (and if possible, comprised of 4 males and 4 females). Any additional pups were selected at random for use in blood-sampling or necropsy procedures. Where 4 males and 4 females were not available on PND 4, additional pups of the opposite sex were used to ensure there were a total of 8 pups. Where the total number of pups in the litter on PND 4 was 8 or fewer, no adjustment was performed. Where practicable, any pups that were found dead or were killed during the postnatal period were sexed and appropriately examined as above. Any externally abnormal decedent pup was preserved in 10% neutral buffered formalin; externally normal pups were discarded.

General In-life Assessments – F1 Animals (Cohorts 1A + Surplus Animals, 1B and 2A)
- Mortality:
All F1 animals At least twice daily (once at the start and once towards the end of the working day) beginning upon arrival through termination/release.
- Pre and Postdose Observations:
All F1 animals. Once predose and up to 4 hours postdose after the end of each group for each sex.
- Detailed Clinical Observations:
All F1 animals. Weekly from weaning, starting on a suitable day within one week of weaning of the majority of litters (weaning on PND 21).
- Individual Body Weights:
All F1 animals. On day of weaning and then weekly, starting on a suitable day within one week of weaning of the majority of litters (beginning of nominal Week 4 of age). For Cohort 1B females only, weekly body weights were continued until pairing for mating and then mated females were weighed on Gestation Day 0, 7, 14 and 20 and Lactation Day 1, 4, 7, 14 and 21. A body weight may also have been taken on Lactation Day 0 for dosing purposes only. Any Lactation Day 0 body
weights were not reported but were maintained in the study data. Pups were weighed individually on PND 1, 4, 7, 14 and 21.
- Food Consumption:
All F1 animals. Excluding surplus animals. Weekly, starting on a suitable day within one week of weaning of the majority of litters (Nominal Week 4 of age).
For Cohort 1B animals, food consumption was recorded until pairing for mating.
1B Mated females: over the periods Gestation Days 0-7, 7-14 and 14-20, and Lactation Days 1-7, 7-14 and 14-21.
1B Males: weekly after mating and re-housing.
- Estrous Cycle Monitoring:
F1 females. Cohort 1A + Surplus Animals. From the day after vaginal patency, continuing until one estrous smear had been identified.
F1 females. Cohort 1A. Smears were also taken for at least 14 consecutive days prior to necropsy, up to and including the day of necropsy.
F1 females. Cohort 1B. From 2 weeks prior to pairing until day of detection of a copulatory plug in situ and/or of sperm in the lavage.
- Water Consumption:
All F1 animals. Regular basis throughout the study.


PARAMETERS EXAMINED
The numbers of live and dead pups born in each litter were recorded as soon as possible after completion of parturition on LD 0. The live pups were counted and examined daily for the presence of milk in the stomach and for any externally visible abnormalities up to and including PND 4 and then on PND 7, 14 and 21. Individual pup weights were taken for each litter on PND 1, 4, 7, 14 and 21. A head count of the pups was performed each day.

Pre-Weaning Physical Development of F1 and F2 Pups:
Ano-genital distance was measured for the pups of both sexes on PND 1. Nipple retention was assessed in males on PND 13.

Assessment of Sexual Maturation Cohorts 1A+ Surplus Animals, 1B and 2A:
Commencing on PND 28, females were examined daily for vaginal opening. The day on which the vagina became open was recorded, as was the body weight on that day.
Commencing on PND 35, males were examined daily for balanopreputial separation. The day on which separation occurred was recorded, as was the body weight on that day.


GROSS EXAMINATION OF DEAD PUPS:
Pups Found Dead or Euthanised Prematurely Before PND 14: Where practicable, these animals were sexed and then checked for the presence of milk in the stomach and for the presence of any externally visible abnormalities. Any externally abnormal pups were fixed in 10% neutral buffered formalin for optional further examination. Externally normal pups were discarded.
Pups Found Dead or Euthanised Prematurely On or After PND 14: These animals were subject to a gross necropsy. An external examination was followed by an inspection of the cranial, thoracic and abdominal contents. Internal sex was also confirmed. Representative samples of any abnormal tissues were taken and fixed in neutral buffered 10% neutral buffered formalin. These carcasses were then discarded.


ASSESSMENT OF DEVELOPMENTAL NEUROTOXICITY:
Perfusion fixation, neuropathology and brain morphometry was performed for all animals. The fixed brains were removed and weighed, and the length and maximum width of the brain was measured for all animals selected for neuropathology.
Brain, dorsal root ganglion, and spinal cords tissues were prepared for neuropathology examination. Neuropathology and brain morphometry were performed.

ASSESSMENT OF DEVELOPMENTAL IMMUNOTOXICITY:
Lymph nodes were weighed to evaluate immunotoxic effects from 10 rats/sex/group from Groups 1-4.
One-half of the spleen was used for immunophenotyping assessment from 10 rats/sex/group from Groups 1-4.
Postmortem examinations (parental animals):
GROSS NECROPSY
- Gross necropsy consisted of a complete necropsy examination, which included evaluation of the carcass and musculoskeletal system; all external surfaces and orifices; cranial cavity and external surfaces of the brain; and thoracic, abdominal, and pelvic cavities with their associated organs and tissues. The reproductive tracts of all F0 females were be examined for signs of implantation, the number of any implantation sites being recorded. The total number of corpora lutea
graviditatis were recorded for each female. For F0 females necropsied on GD 24 as they had failed to produce a litter, the uteri of all non-pregnant females were examined for implantation sites and fixed in 10% neutral buffered formalin. On GD 24, if a female was found to be pregnant, the number and type of any implantation sites were recorded and the total number of corpora lutea graviditatis were recorded also.

HISTOPATHOLOGY / ORGAN WEIGHTS
The following tissues were prepared for microscopic examination and weighed, respectively: Artery, aorta, Body cavity, nasal (Processing at the level of the first
upper molar teeth, to include olfactory bulbs and nasopharynx and/or nasopharyngeal duct), Bone marrow sternum, Bone marrow smear, bone sternum, brain, epididymis, esophagus, eye, dorsal root lumbar ganglion, adrenal gland, clitoral gland, lacrimal gland, harderian gland, mammary gland, parathyroid gland, pituitary gland, preputial gland, prostate gland, submandibular gland, sublingual gland, parotid gland, seminal vesicle gland, thyroid gland, zymbal's gland, Gut-associated lymphoid tissue, heart, kidney, cecum, colon, rectum, larynx, liver, lung, axillary lymph node, mandibular lymph node, mesenteric lymph node, gastrocnemius muscle, quadriceps muscle, optic nerve, sciatic nerve, tibial nerve, ovary, oviduct, pancreas, skin, duodenum ileum, jejunum, spinal cord, spleen, stomach, testis, thymus, tongue, trachea, ureter, urinary bladder, uterus/cervix, vagina, vas deferens.

Immunophenotyping Sample Collection, Processing, and Analysis - F0 Animals and Cohort 1A:
Spleen samples (approximately half of the spleen) were taken from 10 rats/sex/group of the F0 and Cohort 1A animals at necropsy and will be collected into tubes containing RPMI-1640 medium and placed immediately on wet ice until processing. The remainder of the spleen was preserved in 10% neutral buffered formalin.
Upon receipt at the flow cytometry laboratory, the samples were stored on wet ice and were analysed as follows:
The cellular antigens and cell populations identified below were quantified, using specific antibodies against the marker antigens. The samples were analysed using the
Test Facility validated analytical method 996094:

Antigen markers:
CD45+CD3-CD45R+ (B cells)
CD45+CD3+CD45R- (Total T cells)
CD45+CD3+CD45R-CD4+CD8- (T helper cells)
CD45+CD3+CD45R-CD4+CD8- (Cytotoxic T cells)
CD45+CD3-CD45R-CD161+ (Natural killer cells)
Parameters to be Reported - % of lymphocytes
Postmortem examinations (offspring):
Necropsy (F1 and F2 Culled PND 4 and Unselected PND 21 pups):
Culled pups on PND 4 were necropsied. Necropsy consisted of external examination followed by macroscopic examination of the tissues and organs of the cranial, thoracic and abdominal cavities in situ. Samples of any grossly abnormal tissues were preserved in 10% neutral buffered formalin or other appropriate fixative. The carcasses were discarded.
All unselected pups on PND 21 were necropsied. Necropsy consisted of external examination followed by macroscopic examination of the tissues and organs of the cranial, thoracic and abdominal cavities in situ. Representative samples of abnormal tissues were preserved in 10% neutral buffered formalin or other appropriate fixative. For F1 and F2 pups, tissue samples and weights of the following tissues were taken from 10 male and 10 female pups per group and were preserved in 10% neutral buffered formalin or other appropriate fixative: Brain, mammary gland, parathyroid gland, thyroid gland, liver, ovary, spleen, testes, thymus.
Where 10 pups of each sex were not available for organ weight and tissue collection, additional pups of the opposite sex were selected such that 20 animals per group had organ weights and tissues collected.

Necropsy (Cohort 1B Animals):
Animals were subjected to a complete necropsy examination, which included evaluation of the carcass and musculoskeletal system; all external surfaces and orifices; cranial cavity and external surfaces of the brain; and thoracic, abdominal, and pelvic cavities with their associated organs and tissues.
The reproductive tracts of all F1 females were examined for signs of implantation, the number of any implantation sites being recorded. The total number of corpora lutea graviditatis were recorded for each female. For F1 females necropsied on GD 24 as they had failed to produce a litter, the uteri of all non-pregnant females were fixed in 10% neutral buffered formalin after being examined for implantation sites. On GD 24, if a female was found to be pregnant, the number and type of any implantation sites were recorded and the total number of corpora lutea graviditatis were recorded also.


Transcardial Perfusion Fixation of Cohorts 2A and 2B:
All animals were subjected to a limited examination, with special attention being paid to the reproductive organs.
The head (with brain exposed), spinal cord and hindlimbs (Cohort 2A) and head (with brain exposed) (Cohort 2B) was placed in 10% neutral buffered formalin and allowed to fix for at least 7 days. The fixed brains were removed and weighed, and the length and maximum width of the brain was measured for all animals selected for neuropathology. Subsequently, the brain was fixed in 10% neutral buffered formalin together with selected PNS tissues.

Statistics:
Body Weight Changes: Calculated between each scheduled interval
Food Consumption: Calculated between appropriate intervals
Organ Weight Relative to Body Weight: Calculated against the terminal body weight for scheduled
intervals
Organ Weight Relative to Brain Weight: Calculated against the brain weight for scheduled intervals

Descriptive Statistical Analyses:
Means, standard deviations, percentages, numbers, and/or incidences have been reported as appropriate by dataset.

Inferential Statistical Methods:
All statistical tests were conducted at the 5% significance level. All pairwise comparisons were conducted using two sided tests and have been reported at the 1% and 5% levels.
The pairwise comparisons of interest are listed below:
Group 2 vs. Group 1
Group 3 vs. Group 1
Group 4 vs. Group 1
Analyses were performed according to the matrix below when possible but excluded any group with less than 3 observations.

See Table 43 below.

Parametric/Non-parametric:
Levene’s test was used to assess the homogeneity of group variances.
The groups were compared using an overall one-way ANOVA F-test if Levene’s test was not significant or the Kruskal-Wallis test if it was significant. If the overall F-test or Kruskal Wallis test was found to be significant, then pairwise comparisons were conducted using Dunnett’s or Dunn’s test, respectively.

Non-Parametric:
The groups were compared using an overall Kruskal-Wallis test. If the overall Kruskal-Wallis test was found to be significant, then the above pairwise comparisons were conducted using Dunn’s test (equivalent to Wilcoxon Rank-Sum test in Nevis 2012 tables).

Reproductive indices:
Body Weight Changes: Calculated between each scheduled interval
Food Consumption: Calculated between appropriate intervals
Organ Weight Relative to Body Weight: Calculated against the terminal body weight for scheduled
intervals
Organ Weight Relative to Brain Weight: Calculated against the brain weight for scheduled intervals

Descriptive Statistical Analyses:
Means, standard deviations, percentages, numbers, and/or incidences have been reported as appropriate by dataset.

Inferential Statistical Methods:
All statistical tests were conducted at the 5% significance level. All pairwise comparisons were conducted using two sided tests and have been reported at the 1% and 5% levels.
The pairwise comparisons of interest are listed below:
Group 2 vs. Group 1
Group 3 vs. Group 1
Group 4 vs. Group 1
Analyses were performed according to the matrix below when possible but excluded any group with less than 3 observations.

See Table 43 below.

Parametric/Non-parametric:
Levene’s test was used to assess the homogeneity of group variances.
The groups were compared using an overall one-way ANOVA F-test if Levene’s test was not significant or the Kruskal-Wallis test if it was significant. If the overall F-test or Kruskal Wallis test was found to be significant, then pairwise comparisons were conducted using Dunnett’s or Dunn’s test, respectively.

Non-Parametric:
The groups were compared using an overall Kruskal-Wallis test. If the overall Kruskal-Wallis test was found to be significant, then the above pairwise comparisons were conducted using Dunn’s test (equivalent to Wilcoxon Rank-Sum test in Nevis 2012 tables).

Offspring viability indices:
Live Birth Index Percentage of pups born alive: Number of Live Newborn Pups /Number of Newborn Pups x100

Sex Ratio (% males) Percentage of male pups per litter:
Number of Live Male Pups /Total Number of Live Pups x100

Viability Index:
Number of Live Pups on Day 4 Postpartum / Number of Live Newborn Pups x100

Lactation Index:
Number of Live Pups on Day 21 Postpartum /Number of Live Pups on Day 4 (postculling) Postpartum x100

Post-Implantation Loss/Litter:
Number of Implants – Total Newborn Pups /Number of Implants x100

Results and discussion

Results: P0 (first parental generation)

General toxicity (P0)

Clinical signs:
effects observed, treatment-related
Description (incidence and severity):
At 900 mg/kg/day, decreased activity was observed for one male on Day 2 and abnormal, uncoordinated gait for 3 males and 4 females between Days 2 to 4. Decreased activity, abnormal, uncoordinated gait or low carriage were occasionally noted for a further 8 females between Days 5 to 70 and decreased activity on one occasion for each of 2 females during lactation. These findings were considered to be test item-related.
Salivation and/or ploughing (also a chewing action for one female at 900 mg/kg/day) were noted for all animals given ≥300 mg/kg/day and for several animals given 100 mg/kg/day, on multiple days throughout the dosing period. The incidence of these findings was dose level-related.
Animal 4515F (900 mg/kg/day) had abnormal clincal observations of irregular respiratory rate, hunched posture, erect fur, cold to touch, prominent backbone, abnormal gait, decreased activity and red discharge from vagina the day after parturition (LD 1) but these mostly resolved before scheduled termination. The clinical observations were considered to be likely due to the trauma of giving birth rather than being test item-related.
Animal 4525F (900 mg/kg/day) had abnormal clincal observations of irregular/decreased respiratory rate, erect fur, eyes partly closed, decreased activity up to 2-4 hours postdose on LD 7 only. No similar findings were observed for this animal during the dosing period and as a cause could not be determined, these findings were likely related to the dosing procedure.
See Table 1 and Appendix 5.
Mortality:
mortality observed, non-treatment-related
Description (incidence):
None of the unscheduled deaths were considered to be test item-related.
On Study Day 40, approximately 3 hours after dosing, Animal 4510F given 900 mg/kg/day was found with clinical signs of irregular respiratory rate, hunched posture, partly closed eyes, erect fur, uncoordinated and abnormal gait and decreased activity and was euthanised. There were no findings at necropsy or microscopically to suggest the likely cause of death. This animal had not had any abnormal clinical signs before this (apart from salivation and ploughing) and had consistently gained body weight.
Animals 2512F and 4502F given 100 or 900 mg/kg/day respectively, were euthanised on LD 1 or LD 0 and due to the very young age of the pups, they were also euthanised. Animal 2512F had clinical signs of irregular respiratory rate, hunched posture, pale skin and cold to touch, erect fur, uncoordinated and abnormal gait in the afternoon of LD 1. Animal 4502F was found subdued, cold to touch, prostrate/lying on side with decreased respiratory rate approximately 1 hour after dosing on LD 0. There were no findings at necropsy or microscopically to suggest the likely cause of death and both animals had not had any abnormal clinical signs before this (apart from salivation and ploughing for 4502F) and had consistently gained body weight.
See Appendix 4.
Body weight and weight changes:
effects observed, treatment-related
Description (incidence and severity):
For males given 900 mg/kg/day, there was slightly lower body weight gain over the dosing period resulting in male body weight being 3.6% lower than the concurrent control mean value by Day 134. For females given ≥ 100 mg/kg/day, a dose level-related higher body weight gain of up to 10.3% at 900 mg/kg/day compared to controls was noted during the pre-pairing period and this slightly higher body weight was generally maintained throughout gestation and lactation.
There was no test item-related effect on male body weight at 100 or 300 mg/kg/day.
See Table 2, Table 3, Appendix 6, Appendix 7 and Appendix 8.
Food consumption and compound intake (if feeding study):
effects observed, treatment-related
Description (incidence and severity):
There were occasional slightly higher food consumption values of up to 14.4% of controls noted for males at 300 or 900 mg/kg/day up to Day 106. At ≥100 mg/kg/day for males from Days 106 to 134, there was higher food consumption of up to 17.5%. For females given ≥100 mg/kg/day, there was an overall dose level-related higher food consumption of up to 20.6% of control values during the pre-pairing period, and during gestation and lactation, food consumption was higher for females by up to 13.2% at 300 or 900 mg/kg/day only.
See Table 4 and Appendix 9.
Haematological findings:
effects observed, non-treatment-related
Description (incidence and severity):
Haematology
See Table 18 and Appendix 29

Coagulation
See Table 19 and Appendix 30

All differences in haematology or coagulation parameters were considered not to be test item-related based on their small magnitude, inconsistent direction, absence of a dose response, general overlap of individual values with the range of control values and/or were of a magnitude of variation commonly observed in rats.
Clinical biochemistry findings:
effects observed, treatment-related
Description (incidence and severity):
See Table 20 and Appendix 31.
At ≥300 mg/kg/day for males and ≥100 mg/kg/day for females, triglycerides were lower than controls by up to 0.75-fold, and for males only at ≥300 mg/kg/day, cholesterol higher by up to 1.30-fold.
Endocrine findings:
effects observed, non-treatment-related
Description (incidence and severity):
See Table 22, Table 23, Appendix 33, Appendix 34 and Appendix 35.
There were slightly higher TSH levels (up to 3.1-fold of controls) for samples taken on the morning of necropsy in F0 adult males given 300 or 900 mg/kg/day.
There were no test item-related effects for F0 adult males given 100 mg/kg/day or F0 adult females given up to 900 mg/kg/day or for culled pups on PND 4 or unselected pups on PND 21.
There were no test item-related changes to T4 levels at up to 900 mg/kg/day for samples taken on the morning of necropsy for the F0 generation animals, for culled pups on PND 4 or unselected pups on PND 21.
The differences in these parameters were considered not to be test item-related based on their small magnitude, inconsistent direction, absence of a dose response, general overlap of individual values with the range of control and/or were of a magnitude of variation commonly observed in rats.
Urinalysis findings:
effects observed, non-treatment-related
Description (incidence and severity):
See Table 21 and Appendix 32.
All differences in urine parameters were considered not to be test item-related based on their small magnitude, inconsistent direction, absence of a dose response, general overlap of individual values with the range of control and/or were of a magnitude of variation commonly observed in rats.
Immunological findings:
no effects observed
Organ weight findings including organ / body weight ratios:
effects observed, treatment-related
Histopathological findings: non-neoplastic:
effects observed, treatment-related
Description (incidence and severity):
See Table 26, Appendix 37 and Appendix 46
Test item-related Microscopic Findings
In the liver, there was diffuse centrilobular hepatocyte hypertrophy of minimal to mild severity. For males only, this was seen in all dosage groups, whereas for females this was only identified in animals dosed at 900 mg/kg/day. Although this was also found in one control animal (M1021), the incidence was higher in animals treated with Omnirad 184 and followed a dose-related distribution. This finding was therefore considered test item-related in males dosed at ≥ 100 mg/kg/day and in females dosed at 900 mg/kg/day.
The higher liver weights recorded in animals given Omnirad 184, were correlated to the histopathologic findings of centrilobular hypertrophy. This correlation was evident in the males dosed at ≥ 100 mg/kg/day and females dosed at 900 mg/kg/day.
In the glandular portion of the stomach, there was diffuse mucosal hyperplasia with a minimal severity. This was characterised by an increase in the number of goblet cells. For males only, this was seen in at ≥ 300 mg/kg/day, whereas for females this was only identified in animals dosed at 900 mg/kg/day. Although this was also found in one control animal (M1013), the incidence was convincingly higher in animals treated with Omnirad 184 at ≥ 300 mg/kg/day and showed a dose-related distribution. This finding was therefore considered test item-related in males dosed at ≥ 300 mg/kg/day and in females dosed at 900 mg/kg/day.

Additional Microscopic Findings:
In the spleen, there was a slightly higher incidence of increased hematopoietic cells with a minimal severity, primarily in males given Omnirad 184 at a doses of 100 mg/kg/day and 900 mg/kg/day. A slightly higher incidence was seen in females dosed at 900 mg/kg/day only. However, due to the overall low incidences and because this is a common background finding, it was considered not test item-related.
Other microscopic findings were of the nature commonly observed in this strain and age of rat, or occurred at a similar incidence in control and treated animals, and, therefore, were considered not to be test item-related.

Reproductive function / performance (P0)

Reproductive function: oestrous cycle:
no effects observed
Description (incidence and severity):
See Table 5 and Appendix 10.
There was no effect on the number or length of estrous cycles during the 2 weeks before pairing or during the mating period at up to 900 mg/kg/day, compared with controls.
On the morning of necropsy, while the majority of females in all groups were still in diestrus, a few in each group had come back into estrus.
Reproductive function: sperm measures:
no effects observed
Description (incidence and severity):
See Tables 27, Table 28, Table 29, Table 30, Appendix 39, Appendix 40, Appendix 41 and Appendix 42.

Computer Assisted Sperm Analysis:
There were no test item-related effects on the percentage of motile sperm, progressively
motile sperm or straight line velocity at up to 900 mg/kg/day.
F0 control male 1020M control was paired with 1520F which was found not to be pregnant at
scheduled necropsy. The sperm parameters for male 1020M were within the normal ranges.
F0 Animal 2011M given 100 mg/kg/day had a very low sperm motility of 2% at necropsy,
however, the female it had been paired with 2511F, became pregnant and littered
successfully.
At 900 mg/kg/day, F0 Control 1024M was paired with 1524F, and 4010M was paired with
4511F. These females failed to produce litters, however, the sperm parameters for the males
were within the normal ranges.

Sperm Count and Morphological Analysis:
There were no test item-related effects on sperm count or morphology at up to
900 mg/kg/day.

Spermatid Count:
There was no test item-related effect on the number of spermatids at up to 900 mg/kg/day.
Reproductive performance:
no effects observed
Description (incidence and severity):
See Table 6, Table 7, Appendix 11 and Appendix 12.
There were no test item-related effects on male or female mating performance or fertility, or
duration of gestation in females, at up to 900 mg/kg/day.
F0 control 1520F was found not to be pregnant at scheduled necropsy.
F0 Control Animal 1524F and Animal 4511F given 900 mg/kg/day were terminated on
GD 24 as they had failed to produce litters. At necropsy, 1524F had 2 implants (one early
resorption) and 4511F was not pregnant.
F0 Animals 1501F, 1502F, 1506F, 1518F, 1521F (control), 2517F (100 mg/kg/day), 350F2
(300 mg/kg/day) and 4523F (900 mg/kg/day) did not have a positive mating sign during
pairing and therefore gave birth before their expected GD 21.

Effect levels (P0)

Key result
Dose descriptor:
NOAEL
Effect level:
900 mg/kg bw/day
Based on:
test mat.
Sex:
male/female
Basis for effect level:
clinical signs
mortality
body weight and weight gain
food consumption and compound intake
haematology
clinical biochemistry
organ weights and organ / body weight ratios
gross pathology
histopathology: non-neoplastic
reproductive function (oestrous cycle)
reproductive function (sperm measures)
reproductive performance

Results: F1 generation

General toxicity (F1)

Clinical signs:
no effects observed
Description (incidence and severity):
See Table 1 and Appendix 5.
Salivation and/or ploughing were noted for all animals (except Surplus Animal 3650F) given ≥300 mg/kg/day and for a few animals given 100 mg/kg/day, on several days throughout the dosing period. The incidence of these findings was dose level-related.
Surplus Animal 4146M had erect fur on Day 5 postdose and 4147M had irregular respiratory rate, erect fur, eyes partly closed and abnormal gait on Day 4 postdose. Both animals (given 900 mg/kg/day) did not have any similar findings during the dosing period and as a cause could not be determined, these abnormal findings were likely related to the dosing procedure.
For all generations, the other clinical observations recorded were considered not to be test item-related as they were of low incidence, are commonly seen during gestation or lactation, were also seen in some control animals or were not dose level-related.
Mortality / viability:
mortality observed, non-treatment-related
Description (incidence and severity):
See Appendix 4.
Animal 3608F given 300 mg/kg/day had convulsions and died within minutes of receiving its first dose on Day 1. Grossly, the lungs had pale discoloration and abnormal consistency and the trachea had frothy pale content. Microscopically, there was multifocal mixed cell infiltration of the alveolar space of the lung which was considered to correlate with the gross findings in the lung. Overall, these findings were considered to have been related to the dosing procedure and were therefore considered to be the cause of death.
Control Animal 1628F was euthanised on LD 1 because all pups in the litter died. At necropsy, a pale pancreas was noted but no microscopic findings could be correlated and no cause of death was identified. The animal had not had any abnormal clinical signs before this and had consistently gained body weight.
Animal 4634F was found dead in the afternoon of LD 18. At necropsy, in the abdominal cavity, there was dark red fluid accumulation and a soft red mass adherent to the stomach. Microscopically this was correlated with a well demarcated hematocyst of marked severity in relation to hepatic tissue. This finding was considered to be the animal’s cause of death. As it’s pups had not been weaned, they were also euthanised.
Body weight and weight changes:
effects observed, treatment-related
Description (incidence and severity):
See Table 2, Table 3, Appendix 6, Appendix 7 and Appendix 8.
For animals in Cohorts 1A and 1B, there was a general trend of slightly lower body weight gain (up to
7.0%) for males at 900 mg/kg/day compared to controls, but no test item related-effects for males at 100 or 300 mg/kg/day or females at ≥100 mg/kg/day during pre-pairing, gestation or lactation.
In Cohort 2A and for Surplus animals, body weight gain was variable and did not follow any trend.
Food consumption and compound intake (if feeding study):
effects observed, treatment-related
Description (incidence and severity):
See Table 4 and Appendix 9.
There were occasional higher food consumption values of up to 19.6% of controls noted for males at 300 or 900 mg/kg/day, in some F1 generation cohorts. There was a general trend of higher food consumption for females of up to 22.4% higher at 300 or 900 mg/kg/day but during gestation and lactation female food consumption was similar to controls.
Haematological findings:
effects observed, non-treatment-related
Description (incidence and severity):
Haematology
See Table 18 and Appendix 29

Coagulation
See Table 19 and Appendix 30

All differences in haematology or coagulation parameters were considered not to be test item-related based on their small magnitude, inconsistent direction, absence of a dose response, general overlap of individual values with the range of control values and/or were of a magnitude of variation commonly observed in rats.
Clinical biochemistry findings:
effects observed, treatment-related
Description (incidence and severity):
See Table 20 and Appendix 31.
At ≥100 mg/kg/day for males and females, triglycerides were lower than controls by up to 0.62-fold,
and at 900 mg/kg/day only, cholesterol higher by up to 1.25-fold.
For both generations, all other differences in clinical chemistry parameters were considered not to be test item-related based on their small magnitude, inconsistent direction, absence of a dose response, general overlap of individual values with the range of control and/or were of a magnitude of variation commonly observed in rats.
Urinalysis findings:
effects observed, non-treatment-related
Description (incidence and severity):
See Table 21 and Appendix 32.
All differences in urine parameters were considered not to be test item-related
based on their small magnitude, inconsistent direction, absence of a dose response, general overlap of individual values with the range of control and/or were of a magnitude of variation commonly observed in rats.
Sexual maturation:
no effects observed
Description (incidence and severity):
See Table 13, Appendix 23 and Appendix 24.
There was considered to be no effect on sexual maturation for F1 males or females at up to 900 mg/kg/day.
Anogenital distance (AGD):
no effects observed
Description (incidence and severity):
See Table 12 and Appendix 21.
There was no effect on anogenital distance or normalised anogenital distance for males or females at up to 900 mg/kg/day.
Nipple retention in male pups:
no effects observed
Description (incidence and severity):
See Appendix 22.
On PND 13, there was no effect on nipple retention in male pups i.e. no nipples were present, at dose levels up to 900 mg/kg/day.
Organ weight findings including organ / body weight ratios:
effects observed, treatment-related
Description (incidence and severity):
See Table 24 and Appendix 36.
F1 Cohort 1A
Test item-related Organ Weight Differences:
Liver weights in animals dosed at ≥ 300 mg/kg/day were statistically significantly higher than the controls and showed a dose-related response. In males, liver weights were up to ~24% higher and in females up to ~29% higher (relative to body weight at 900 mg/kg/day). This was therefore considered test item-related.
Kidney weights in animals dosed at ≥ 300 mg/kg/day in males and at ≥ 100 mg/kg/day in females, were statistically significantly higher than the controls and showed a dose related response. In males, kidney weights were up to ~23% higher and in females up to ~16% higher (relative to body weight at 900 mg/kg/day). Although this was not correlated to any microscopic findings, this was considered test item-related in view of the similar weight differences recorded in the F0 and F1 cohort 1B animals.

Additional Organ Weight Differences:
Testis weights relative to body weight were statistically significantly higher by ~11% only at 900 mg/kg/day compared to the controls. However, males dosed at 900 mg/kg/day had a lower terminal body weight and in view of the minor weight difference and no microscopic findings that could be correlated. This was therefore considered incidental.
Ovary weights were statistically significantly higher by ~19% (relative to body weight at 300 mg/kg/day). However, no histologic findings could be correlated to this finding and no similar differences were found in any other generation or cohort of animals, so this was considered incidental.
Thyroid/parathyroid gland weights were statistically significantly higher by ~21% (relative to body weight) in female animals dosed at 900 mg/kg/day, compared to controls. However, no histologic findings could be correlated to this finding, thyroid hormone was not increased in a noteworthy fashion, and no similar differences were found in any other generation or cohort of animals, so this was considered incidental.
Seminal vesicle weights were statistically significantly higher by ~22% (relative to body weight) in animals dosed at 900 mg/kg/day, compared to controls. However, no histologic findings could be correlated to this finding, so this was considered not test item-related.
There were individual organ weight values that were different from their respective controls. There were, however, no patterns or correlating data to suggest these values were test item-related.

F1 Cohort 1B
Test item-related Organ Weight Differences:
Liver weights in animals dosed at ≥ 300 mg/kg/day were statistically significantly higher than the controls and showed a dose-related response. In males, liver weights were up to ~24% higher and in females up to ~29% higher (relative to body weight at 900 mg/kg/day). This was therefore considered test item-related.
Kidney weights in animals dosed at ≥ 300 mg/kg/day in males and at ≥ 100 mg/kg/day in females, were statistically significantly higher than the controls and showed a dose related response. In males, kidney weights were up to ~23% higher and in females up to ~16% higher (relative to body weight at 900 mg/kg/day). Although this was not correlated to any microscopic findings, this was considered test item-related in view of the similar weight differences recorded in the F0 and F1 cohort 1B animals.
Additional Organ Weight Differences:
Testis weights relative to body weight were statistically significantly higher by ~11% only at 900 mg/kg/day compared to the controls. However, males dosed at 900 mg/kg/day had a lower terminal body weight and in view of the minor weight difference and no microscopic findings that could be correlated. This was therefore considered incidental.
Ovary weights were statistically significantly higher by ~19% (relative to body weight at 300 mg/kg/day). However, no histologic findings could be correlated to this finding and no similar differences were found in any other generation or cohort of animals, so this was considered incidental.
Thyroid/parathyroid gland weights were statistically significantly higher by ~21% (relative to body
weight) in female animals dosed at 900 mg/kg/day, compared to controls. However, no histologic findings could be correlated to this finding, thyroid hormone was not increased in a noteworthy fashion, and no similar differences were found in any other generation or cohort of animals, so this was considered incidental.
Seminal vesicle weights were statistically significantly higher by ~22% (relative to body weight) in
animals dosed at 900 mg/kg/day, compared to controls. However, no histologic findings could be correlated to this finding, so this was considered not test item-related.
(Cohort 2A, 2B, F1 unselected pups, F2 unselected pups):
There were no test item-related organ weight differences.
Gross pathological findings:
no effects observed
Description (incidence and severity):
See Appendix 38.
Scheduled Euthanasia Animals - F1 Animals:
There were no test item-related gross findings.
Gross findings observed were of the nature commonly observed in this strain and age of rat, or occurred at a similar incidence in control and treated animals, and, therefore, were considered not to be test item-related.

Scheduled Euthanasia Animals (F1 Unselected pups):
There were no test item-related gross pathology findings.
The only abnormality noted was a dark red focus on the thymus in one male pup. Although the parents of these pups had been given Omnirad 184 at a dose of 900 mg/kg/day, this finding was considered not to be test item-related because it is a common agonal change.
Histopathological findings:
effects observed, treatment-related
Description (incidence and severity):
See Table 26, Appendix 37 and Appendix 46.
(Cohort 1A)
In the liver, there was diffuse centrilobular hypertrophy of minimal to mild severity. For males only,
this was seen in all dosage groups, whereas for females this was only identified in animals dosed at ≥ 300 mg/kg/day. The incidence followed a dose-related distribution and was therefore considered test item-related in males dosed at ≥ 100 mg/kg/day and in females dosed at ≥ 300 mg/kg/day.
The higher liver weights recorded in animals given Omnirad 184 were correlated to the histopathologic findings of centrilobular hypertrophy. This correlation was evident in males dosed at ≥ 100 mg/kg/day and females at ≥ 300 mg/kg/day.
In the glandular portion of the stomach, there was diffuse mucosal hyperplasia with a minimal severity. This was characterised by an increase in the number of goblet cells. For both females and males, this was seen at ≥ 300 mg/kg/day. Although at a low incidence, this finding was only seen in animals treated with Omnirad 184 and in view of similar findings seen in the F0 generation, this finding was considered test item-related in males and females dosed at ≥ 300 mg/kg/day.
Additional Microscopic Findings:
In the kidneys, there was a higher incidence of basophilic tubules with a minimal to mild severity, in male animals given Omnirad 184 at a dose of 900 mg/kg/day. This was however a small difference and although the males from F0 and F1 cohort 1B showed a similar difference, it was even less apparent at the highest dose level (900 mg/kg/day). Due to the fact that this lesion is a common background finding and occurred at a low severity, this was considered incidental.
In the liver of three animals given Omnirad 184 (two females dosed at 100 mg/kg/day and one female dosed at 300 mg/kg/day), there was focal necrosis with subacute inflammation in the medial lobe parenchyma, with a mild severity. Due to the low incidence, this finding was considered not test item related.

(Cohort 1B)
In the liver, there was diffuse centrilobular hypertrophy of minimal to mild severity. For males only, this was seen in all dosage groups, whereas for females this was only identified in animals dosed at ≥ 300 mg/kg/day. The overall incidence followed a dose related distribution and was therefore considered test item-related in males dosed at ≥ 100 mg/kg/day and in females dosed at ≥ 300 mg/kg/day.
The higher liver weights recorded in animals given Omnirad 184, was correlated to the histopathologic findings of centrilobular hypertrophy. This correlation was evident in males dosed at ≥ 100 mg/kg/day and females at ≥ 300 mg/kg/day. There were no test item-related microscopic findings in the reproductive tissues.

(F1 Cohort 2A and 2B)
There were no test item-related microscopic findings. Microscopic findings occurred at similar incidences in control and treated animals, and, therefore, were considered not to be test item-related.
Other effects:
no effects observed
Description (incidence and severity):
Acoustic startle:
See Table 14 and Appendix 25.
There were no test item-related effects on maximum response amplitude, average response amplitude or time to maximum response in males or females at up to 900 mg/kg/day.

Estrous Cycles:
See Table 5 and Appendix 10.
The number of days after vaginal opening before an estrus occurred was comparable with controls at all test item dose levels. There was no effect on the number or length of estrous cycles during the 2 weeks before necropsy at up to 900 mg/kg/day, compared with controls.
There was no effect on the number or length of estrous cycles during the 2 weeks before pairing or during the mating period at up to 900 mg/kg/day, compared with controls.

Mating Performance, Fertility and Duration of Gestation:
See Table 6, Table 7, Appendix 11 and Appendix 12.
The following Cohort 1B females were euthanised on GD 24 as they had failed to produce litters and were found not to be pregnant; 1629F (control), 2624F, 2627F, 2628F and 2637F (100 mg/kg/day) and 3629F (300 mg/kg/day).
See table 22, table 24, and table 26

Pre- and Post-Implantation Loss:
See Table 8 and Appendix 13.
There were no test item-related effects on number of corpora lutea, implant counts, pre- or post-implantation loss or total number of pups born at up to 900 mg/kg/day.

Developmental immunotoxicity (F1)

Developmental immunotoxicity:
no effects observed
Description (incidence and severity):
Splenic Immunophenotyping Evaluation:
See Appendix 44.
The results demonstrated no test item, dose-dependent or sex-dependent effects on any of the
immune cell populations in either the F0 adults rats analysed following administration of
Omnirad 184 by oral gavage, or their resulting offspring.

Effect levels (F1)

Key result
Dose descriptor:
NOAEL
Generation:
F1
Effect level:
900 mg/kg bw/day
Based on:
test mat.
Sex:
male/female
Basis for effect level:
sexual maturation
clinical signs
mortality
body weight and weight gain
food consumption and compound intake
haematology
clinical biochemistry
urinalysis
organ weights and organ / body weight ratios
gross pathology
histopathology: non-neoplastic
developmental immunotoxicity

Results: F2 generation

General toxicity (F2)

Clinical signs:
not examined
Mortality / viability:
no mortality observed
Description (incidence and severity):
There were no unscheduled deaths for F2 generation animals.
Body weight and weight changes:
not examined
Food consumption and compound intake (if feeding study):
not examined
Haematological findings:
not examined
Clinical biochemistry findings:
not examined
Urinalysis findings:
not examined
Sexual maturation:
not examined
Anogenital distance (AGD):
no effects observed
Description (incidence and severity):
There was no effect on anogenital distance or normalised anogenital distance for males or
females at up to 900 mg/kg/day.
Nipple retention in male pups:
not examined
Organ weight findings including organ / body weight ratios:
no effects observed
Description (incidence and severity):
See Table 24 and Appendix 36.
There were no test item-related organ weight differences.
Gross pathological findings:
no effects observed
Description (incidence and severity):
See Appendix 38.
There were no test item-related gross pathology findings.
Abnormalities noted at necropsy included a dark red focus on the thymus in one female pup. Although the parents of these pups had been given Omnirad 184 at a dose of 100 mg/kg/day, this finding was considered not to be test item-related because it is a common agonal change. In the liver of one male and three female pups whose parents had been treated with Omnirad 184 at a dose of 100 mg/kg/day there was mottled discoloration, abnormal appearance and a pale nodule on the medial lobe. Due to the low incidence and because no other animals were found with these gross lesions, they were considered not test item-related.
Histopathological findings:
not examined

Developmental neurotoxicity (F2)

Behaviour (functional findings):
not examined

Developmental immunotoxicity (F2)

Developmental immunotoxicity:
not examined

Effect levels (F2)

Key result
Dose descriptor:
NOAEL
Generation:
F2
Effect level:
900 mg/kg bw/day
Based on:
test mat.
Sex:
male/female
Basis for effect level:
viability
sexual maturation
mortality
organ weights and organ / body weight ratios
gross pathology

Overall reproductive toxicity

Key result
Reproductive effects observed:
no
Lowest effective dose / conc.:
900 mg/kg bw/day
Treatment related:
no

Any other information on results incl. tables

Table 2 - Summary of Body Weights (g)


F0 Males



































































































































































































Sex: Male Day(s) Relative to Start Date
-71815222936
0 mg/kg/dayMean  124.9176.6225.1268.9303.1333355.5
 Group 1SD  16.618.52123.824.726.327.3
 N  25    - 25    - 25    - 25    - 25    - 25    - 25    - 
          
 100  mg/kg/dayMean  123.8174.3225.7270.7305.6333.2355.7
 Group 2SD  16.318.823.425.226.428.530.7
 N  25252525252525
 %Diff  -0.9-1.30.30.70.80.10.1
 300  mg/kg/dayMean  121171.9222.1268.3302.8332.4355.9
 Group 3SD  15.118.92123.122.124.626.8
 N  25252525252525
 %Diff  -3.1-2.7-1.3-0.2-0.1-0.20.1
 900  mg/kg/dayMean  124.9178.2230.5277.9310.1334.8352.2
 Group 4SD  11.614.218.921.924.125.728.3
 N  25252525252525
 %Diff  00.92.43.42.30.5-0.9

 



































































































































































































Sex: Male Day(s) Relative to Start Date
43505764717885
0 mg/kg/dayMean  374.2389406.9417.8427.7428435.6
Group 1SD  30.231.633.73534.933.234.4
 N  25    - 25    - 25    - 25    - 25    - 25    - 25    - 
          
 100  mg/kg/dayMean  374.4392.8408417.3423.9427.8437.4
Group 2SD  32.834.936.135.536.737.437.7
 N   25252525252525
 %Diff 0.110.3-0.1-0.9-0.10.4
 300  mg/kg/dayMean  373.7390.5404.8415.6424426.6436.5
Group 3SD  29.832.233.233.836.936.438.4
 N25252525252525
 %Diff -0.10.4-0.5-0.5-0.9-0.30.2
 900  mg/kg/dayMean  365.9378.8393.4403.8411.4415.7423.1
Group 4SD  29.73233.134.735.534.636.7
 N25252525252525
 %Diff -2.2-2.6-3.3-3.3-3.8-2.9-2.9

 



































































































































































































Sex: Male Day(s) Relative to Start Date
9299106113120127134
0 mg/kg/dayMean  441.8453.6466.3472.8483.5489.2496.7
Group 1SD  35.936.137.837.838.93941.7
 N  25    - 25    - 25    - 25    - 25    - 25    - 25    - 
          
 100  mg/kg/dayMean  441.6450.2462.2470.4478487.7492.6
Group 2SD  37.137.639.740.741.143.845.4
 N   25252525252525
 %Diff -0.1-0.7-0.9-0.5-1.1-0.3-0.8
 300  mg/kg/dayMean  446.7452.6463.6475.6483.9492.7496.6
Group 3SD  40.742.344.74749.750.953.1
 N   25252525252525
 %Diff 1.1-0.2-0.60.60.10.70
 900  mg/kg/dayMean  434.1442.2449.5458.4464.4475.5478.6
Group 4SD  3839.840.340.740.742.544.8
 N   25252525252525
 %Diff -1.7-2.5-3.6-3-3.9-2.8-3.6

 
















































































































































Sex: Male Day(s) Relative to Start Date
137138141142
0 mg/kg/dayMean  507.9501.8493.1473.4
Group 1SD  58.233.536.143.3
 N  8    - 6    - 11    - 5    - 
       
 100  mg/kg/dayMean  488512.6490.8477.7
Group 2SD  46.539.650.563.9
 N   57137
 %Diff -3.92.1-0.50.9
 300  mg/kg/dayMean  458534.4491494.8
Group 3SD  26.457.846.452.9
 N   48138
 %Diff -9.86.5-0.44.5
 900  mg/kg/dayMean  473.8494.5480.2471.2
Group 4SD  41.827.554.165.1
 N   84135
 %Diff -6.7-1.5-2.6-0.5

 


Table 2 - Summary of Body Weights (g)


F0 Females - Prior to mating









































































































































































































Sex: Female    Day(s) Relative to Start Date   
-71815222936
0 mg/kg/dayMean  72.9110.5136.4155.4173.6188.9197.4
Group 1SD  13.513.91413.813.313.914.6
 N  25    - 25    - 25    - 25    - 25    - 25    - 25    - 
          
 100  mg/kg/dayMean  76114.9140.8162181.1195.7206.3
Group 2SD  10.211.111.613.514.414.516.5
 N   25252525252525
  %Diff4.23.93.34.34.33.64.5
 300  mg/kg/dayMean  74.9114.2142.2162.9183.5198.8209.1*
Group 3SD  12.411.110.612.312.814.715.6
 N   25252525252525
  %Diff2.83.44.34.95.75.25.9
 900  mg/kg/dayMean  77.2116.4147.2*171.3**191.6**208.2**214.9**
Group 4SD  14.114.315.317.217.417.918.2
 N   25252525252525
  %Diff5.95.37.910.310.310.28.9

 





































































































































































Sex: Female   Day(s) Relative to Start Date  
4350576471
0 mg/kg/dayMean  207.3214.6222.2226.2230.5
Group 1SD  14.913.114.814.415.6
 N  25    - 25    - 25    - 25    - 25    - 
        
 100  mg/kg/dayMean  216225.6229.6232.8240.8
Group 2SD  17.118.41920.520.5
 N   2525252525
  %Diff4.25.23.32.94.5
 300  mg/kg/dayMean  217.2227.9*234.7*238.4245.9*
Group 3SD  16.518.619.418.919
 N   2525252525
  %Diff4.86.25.65.46.7
 900  mg/kg/dayMean  227.0**233.0**242.0**244.5**250.5**
Group 4SD  17.217.518.61919.7
 N   2424242424
  %Diff9.58.68.98.18.7

Anova & Dunnett: * = p ≤ 0.05; ** = p ≤ 0.01


 


Table 2 - Summary of Body Weights (g)


F0 Females - Gestation


















































































































































Sex: Female  Day(s) Relative to Mating (Litter: A) 
071420
0 mg/kg/dayMean  222.8246.1269.9325.9
Group 1SD  14.513.516.324
 N  18    - 18    - 18    - 18    - 
       
 100  mg/kg/dayMean  237.2*259.5282.3338.5
Group 2SD  20.120.522.931.2
 N   24242424
  %Diff6.55.54.63.9
 300  mg/kg/dayMean  242.1**262.4*284.2343.6
Group 3SD  20.522.424.628.1
 N   24242424
  %Diff8.66.65.35.4
 900  mg/kg/dayMean  243.7**264.5*291.1*352.5*
Group 4SD  17.221.123.434.4
 N   22222222
  %Diff9.47.57.98.2

Anova & Dunnett: * = p ≤ 0.05; ** = p ≤ 0.01


 


Table 2 - Summary of Body Weights (g)


F0 Females - Lactation

























































































































































Group / 14Day1421
Sex7
1FMean261.7274275.1282.8280.1
  SD19.120.819.321.216.6
 N2322232323
 Mean268.2280.9283.3290.9288
2F SD22.122.62323.621.7
 N2324242424
 %Diff G12.52.532.92.8
 Mean270.8282.7285.6294.8291
  SD25.72524.423.419.9
3FN2525252525
 %Diff G13.53.23.84.23.9
 Mean271.6286.4292.2302.6a300.1b
4F SD23.42424.825.725.2
 N2222222222
 %Diff G13.84.56.277.1

Significantly different from control group 1 value :a=p≤0.05,b=p≤0.01 (Dunnett)


 


Table 2 - Summary of Body Weights (g)


F1 Animals - Cohort 1A









































































































































































































Sex: Male    Day(s) Relative to Start Date   
1 [G]2 [I]6 [G]13 [G]20 [G]27 [G]34 [G]
0 mg/kg/dayMean 56-71.8119.9172.1219.5269.6
Group 1SD 5.1-8.812.215.819.723.4
 20    --20    -20    -20    -20    -20    -
   -     
 100  mg/kg/dayMean 56.361.0n70.8116.4166.9213.7261.2
Group 2SD 6.66.6913.616.520.526.4
 N  193    -1820202020
 %Diff 0.7 -1.4-2.9-3-2.6-3.1
 300  mg/kg/dayMean 57.976.0n76.8126.3179.8227276.8
Group 3SD 4.9-9.31416.919.624.9
 N  201    -1920202020
 %Diff 3.5 6.95.44.53.42.7
 900  mg/kg/dayMean 54.759.0n 4.370.6115.8167.5212.9258.5
Group 4SD 5.56    -8.212.516.521.725.5
 N  15 1920202020
 %Diff -2.2 -1.6-3.4-2.7-3-4.1

[G] - Anova & Dunnett [I] - n - Inappropriate for statistics


 









































































































































































































Sex: Male    Day(s) Relative to Start Date   
41485562686970
0 mg/kg/dayMean 309.2343.6368.2390.7408.1394405.5
Group 1SD 26.731.5333740.142.240.7
 20    -20    -20    -20    -14    -6    -20    -
         
 100  mg/kg/dayMean 300.1335.3360.5382.8393408.2401.6
Group 2SD 29.432.635.736.535.342.734.3
 N  2020202016616
 %Diff -2.9-2.4-2.1-2-3.73.6-1
 300  mg/kg/dayMean 317.3349.4376.1396.8411.1445.9411.1
Group 3SD 283337.638.746.236.636.1
 N  2020202014719
 %Diff 2.61.72.11.60.813.21.4
 900  mg/kg/dayMean 299.2325.5346.4363.5386.8381.7382.7
Group 4SD 2932.137.54040.648.238.9
 N  20202020141115
 %Diff -3.2-5.3-5.9-7-5.2-3.1-5.6

 


































Sex: Male



 



Day(s) Relative to Start Date



71



 0


 mg/kg/day


 


 Group 1



Mean 


SD 



 



-


-


-


-



 100  mg/kg/day


 


 Group 2



Mean 


SD 


N  %Diff 



447.0n


69.3


2    -



 300  mg/kg/day


 


 Group 3



Mean 


SD 


N  %Diff 



-


-


-


-



 900  mg/kg/day


 


 Group 4



Mean 


SD 


N  %Diff 



-


-


-


-



 






































































Sex: Female



 



 



 



 



Day(s) Relative to Start Date



 



 



 



1 [G]



2 [I]



6 [G]



13 [G]



20 [G]



27 [G]



34 [G]



 0


 mg/kg/day


 


 Group 1



Mean 


SD 



 



52.6


4.9


20    -



-


-


-


-



66.0


7.8


20    -



103.8


9.7


20    -



136.5


10.7


20    -



157.2


10.2


20    -



176.5


12.3


20    -



 100  mg/kg/day


 


 Group 2



Mean 


SD 


N  %Diff 



55.3


4.9


19   


5.3



58.0n


3.0


3    -



68.9


5.7


18   


4.5



106.1


9.4


20   


2.2



138.3


11.9


20   


1.3



159.7


14.1


20   


1.6



178.8


16.8


20   


1.3



 300  mg/kg/day


 


 Group 3



Mean 


SD 


N  %Diff 



55.0


5.0


21   


4.7



71.0n


-


1    -



69.4


8.9


19   


5.2



105.5


11.3


20   


1.7



138.6


11.6


20   


1.5



160.2


12.3


20   


1.9



179.9


13.8


20   


1.9



 900  mg/kg/day


 


 Group 4



Mean 


SD 


N  %Diff 



52.1


4.8


15   


-0.8



56.3n 6.0


7    -



66.1


8.2


18   


0.2



103.2


10.6


20   


-0.6



138.6


12.8


20   


1.5



159.9


15.9


20   


1.7



182.0


15.5


20   


3.1



[G] - Anova & Dunnett


 






































































Sex: Female



 



 



 



 



Day(s) Relative to Start Date



 



 



 



41 [G]



48 [G]



55 [G]



62 [G]



68 [G]



69 [G]



70 [I]



 0


 mg/kg/day


 


 Group 1



Mean 


SD 



 



189.0


12.1


20    -



203.2


14.4


20    -



212.2


15.5


20    -



221.0


18.8


20    -



231.6


18.8


13    -



228.1


18.9


7    -



245.3n


11.0


3    -



 100  mg/kg/day


 


 Group 2



Mean 


SD 


N  %Diff 



192.4


18.5


20   


1.8



205.7


17.3


20   


1.2



214.1


18.0


20   


0.9



224.2


20.5


20   


1.4



235.2


22.6


17   


1.6



216.7


6.8


3   


-5.0



-


-


-


-



 300  mg/kg/day


 


 Group 3



Mean 


SD 


N  %Diff 



194.4


13.6


20   


2.8



206.5


14.6


20   


1.6



216.4


16.9


20   


2.0



226.1


17.7


20   


2.3



231.5


16.3


14   


0.0



242.3


12.6


6   


6.2



220.0n


-


1    -10.3



 900  mg/kg/day


 


 Group 4



Mean 


SD 


N  %Diff 



201.6


17.1


20   


6.7



209.2


17.6


20   


3.0



218.0


17.7


20   


2.7



226.5


18.3


20   


2.5



237.5


19.1


15   


2.6



235.6


26.0


5   


3.3



-


-


-


-



[G] - Anova & Dunnett


 


















































Sex: Female



 



Day(s) Relative to Start Date



 



71 [G]



72 [I]



73 [G]



74 [G]



 0


 mg/kg/day


 


 Group 1



Mean 


SD 



 



226.9


16.6


8    -



-


-


-


-



228.6


21.1


5    -



228.8


17.9


4    -



 100  mg/kg/day


 


 Group 2



Mean 


SD 


N  %Diff 



228.3


29.4


8   


0.6



223.7n


12.2


3    -



233.8


11.4


5   


2.3



233.8


25.4


4   


2.2



 300  mg/kg/day


 


 Group 3



Mean 


SD 


N  %Diff 



233.1


15.5


9   


2.7



268.0n


-


1    -



225.0n


11.3


2   


-1.6



237.7


20.5


7   


3.9



 900  mg/kg/day


 


 Group 4



Mean 


SD 


N  %Diff 



235.0


23.9


10   


3.6



247.0n


11.3


2    -



232.8


19.2


6   


1.9



235.0n


25.5


2   


2.7



[G] - Anova & Dunnett: n - Inappropriate for statistics


 


Table 2 - Summary of Body Weights (g)


F1 Animals - Cohort 1B Males






































































Sex: Male



 



 



 



 



Day(s) Relative to Start Date



 



 



 



1 [G]



2 [I]



6 [G]



13 [G]



20 [G]



27 [G]



34 [G]



 0


 mg/kg/day


 


 Group 1



Mean 


SD 



 



54.6


5.4


20    -



-


-


-


-



70.5


8.5


20    -



116.5


12.0


20    -



167.7


13.3


20    -



212.9


17.3


20    -



261.7


20.9


20    -



 100  mg/kg/day


 


 Group 2



Mean 


SD 


N  %Diff 



57.4


5.2


19   


5.1



60.0n


8.5


3    -



72.9


8.7


18   


3.5



117.5


14.9


20   


0.8



166.8


20.5


20   


-0.6



211.2


25.9


20   


-0.8



258.5


29.7


20   


-1.2



 300  mg/kg/day


 


 Group 3



Mean 


SD 


N  %Diff 



56.8


5.7


20   


4.0



70.0n


-


1    -



74.9


10.8


19   


6.2



124.1


15.9


20   


6.5



175.7


18.3


20   


4.7



222.6


21.4


20   


4.6



271.2


25.8


20   


3.6



 900  mg/kg/day


 


 Group 4



Mean 


SD 


N  %Diff 



54.5


4.5


15   


-0.1



59.3n 5.9


7    -



69.9


8.7


18   


-0.8



115.4


10.8


20   


-0.9



166.8


12.3


20   


-0.5



212.2


15.2


20   


-0.3



258.6


18.7


20   


-1.2



 






































































Sex: Male



 



 



 



 



Day(s) Relative to Start Date



 



 



 



41



48



55



62



69



76



83



 0


 mg/kg/day


 


 Group 1



Mean 


SD 



 



300.5


25.6


20    -



334.6


30.5


20    -



358.4


34.8


20    -



381.5


38.4


20    -



395.5


41.5


20    -



410.3


44.0


20    -



421.2


46.7


20    -



 100  mg/kg/day


 


 Group 2



Mean 


SD 


N  %Diff 



297.5


32.5


20   


-1.0



330.9


34.9


20   


-1.1



357.3


37.9


20   


-0.3



379.6


39.8


20   


-0.5



399.5


39.4


20   


1.0



414.5


41.7


20   


1.0



429.9


42.0


20   


2.1



 300  mg/kg/day


 


 Group 3



Mean 


SD 


N  %Diff 



310.2


26.4


20   


3.2



342.6


29.7


20   


2.4



370.5


31.5


20   


3.4



393.0


33.5


20   


3.0



413.3


33.9


20   


4.5



427.1


37.4


20   


4.1



438.9


39.0


20   


4.2



 900  mg/kg/day


 


 Group 4



Mean 


SD 


N  %Diff 



297.9


23.2


20   


-0.8



324.7


26.1


20   


-2.9



348.3


30.2


20   


-2.8



365.3


33.5


20   


-4.2



383.9


36.0


20   


-2.9



399.6


35.5


20   


-2.6



410.4


40.6


20   


-2.6



 






































































Sex: Male



 



 



 



 



Day(s) Relative to Start Date



 



 



 



90



97



104



111



118



125



128



 0


 mg/kg/day


 


 Group 1



Mean 


SD 



 



431.7


46.1


20    -



434.7


44.9


20    -



441.9


43.5


20    -



453.5


48.3


20    -



464.3


49.9


20    -



473.8


52.6


20    -



484.1


58.9


8    -



 100  mg/kg/day


 


 Group 2



Mean 


SD 


N  %Diff 



440.9


44.6


20   


2.1



444.3


45.0


20   


2.2



452.5


46.6


20   


2.4



463.5


46.1


20   


2.2



477.7


47.1


20   


2.9



492.3


48.2


20   


3.9



494.2


58.5


11   


2.1



 300  mg/kg/day


 


 Group 3



Mean 


SD 


N  %Diff 



448.1


41.2


20   


3.8



452.0


38.8


20   


4.0



456.2


39.7


20   


3.2



471.1


40.7


20   


3.9



483.8


43.3


20   


4.2



494.5


45.3


20   


4.4



515.1


42.9


9   


6.4



 900  mg/kg/day


 


 Group 4



Mean 


SD 


N  %Diff 



415.5


40.8


20   


-3.8



424.9


41.7


20   


-2.3



428.4


41.3


20   


-3.1



442.0


42.1


20   


-2.5



451.7


42.8


20   


-2.7



460.8


41.2


20   


-2.8



463.5


45.3


12   


-4.3



 














































Sex: Male



 



 



Day(s) Relative to Start Date



 



129 [G]



130 [G]



131 [I]



 0


 mg/kg/day


 


 Group 1



Mean 


SD 



 



465.0


62.2


6    -



475.3


45.0


9    -



-


-


-


-



 100  mg/kg/day


 


 Group 2



Mean 


SD 


N  %Diff 



504.9


36.5


7   


8.6



502.0


45.0


7   


5.6



-


-


-


-



 300  mg/kg/day


 


 Group 3



Mean 


SD 


N  %Diff 



514.8


41.9


5   


10.7



471.0


41.8


8   


-0.9



485.5n


36.8


4    -



 900  mg/kg/day


 


 Group 4



Mean 


SD 


N  %Diff 



476.3


43.3


8   


2.4



436.3


10.1


3   


-8.2



529.0n


-


1    -



 


Table 2 - Summary of Body Weights (g)


F1 Animals - Cohort 1B Females - Prior to Mating






































































Sex: Female



 



 



 



 



Day(s) Relative to Start Date



 



 



 



1 [G]



2 [I]



6 [G]



13 [G]



20 [G]



27 [G]



34 [G]



 0


 mg/kg/day


 


 Group 1



Mean 


SD 



 



52.6


5.1


20    -



-


-


-


-



66.2


7.1


20    -



103.6


9.0


20    -



136.1


9.5


20    -



156.9


10.6


20    -



174.8


12.3


20    -



 100  mg/kg/day


 


 Group 2



Mean 


SD 


N  %Diff 



53.8


7.9


19   


2.4



57.3n


4.0


3    -



66.6


10.4


18   


0.7



103.4


13.9


20   


-0.1



133.7


13.8


20   


-1.7



155.3


14.0


20   


-1.0



173.5


14.1


20   


-0.7



 300  mg/kg/day


 


 Group 3



Mean 


SD 


N  %Diff 



55.7


4.4


20   


6.0



68.0n


-


1    -



71.6


7.5


19   


8.3



109.3


8.1


20   


5.5



143.2


9.6


20   


5.3



164.1


10.0


20   


4.6



180.9


11.8


20   


3.5



 900  mg/kg/day


 


 Group 4



Mean 


SD 


N  %Diff 



51.3


4.3


15   


-2.3



56.3n 6.6


7    -



64.6


7.4


18   


-2.4



102.8


10.3


20   


-0.7



136.6


9.8


20   


0.4



157.6


13.1


20   


0.4



176.8


13.7


20   


1.1



 






































































Sex: Female



 



 



 



 



Day(s) Relative to Start Date



 



 



 



41



48



55



62



69



76



83



 0


 mg/kg/day


 


 Group 1



Mean 


SD 



 



189.6


11.3


20    -



204.2


11.0


20    -



210.9


11.8


20    -



218.6


12.6


20    -



227.6


11.8


20    -



232.5


11.5


20    -



236.2


12.5


20    -



 100  mg/kg/day


 


 Group 2



Mean 


SD 


N  %Diff 



188.1


15.8


20   


-0.8



200.3


15.1


20   


-1.9



209.8


15.4


20   


-0.5



218.7


14.7


20   


0.1



226.3


16.5


20   


-0.6



230.7


18.2


20   


-0.8



232.7


17.3


20   


-1.5



 300  mg/kg/day


 


 Group 3



Mean 


SD 


N  %Diff 



199.2


12.4


20   


5.1



211.3


13.2


20   


3.5



220.1


14.6


20   


4.4



227.0


15.2


20   


3.9



236.9


15.5


20   


4.1



243.1


16.4


20   


4.5



246.7


17.5


20   


4.5



 900  mg/kg/day


 


 Group 4



Mean 


SD 


N  %Diff 



195.6


14.1


20   


3.2



203.7


14.7


20   


-0.2



212.2


15.4


20   


0.6



219.5


16.9


20   


0.4



231.0


16.5


20   


1.5



236.4


18.1


20   


1.7



237.4


17.1


20   


0.5



 


































Sex: Female



 



Day(s) Relative to Start Date



90



 0


 mg/kg/day


 


 Group 1



Mean 


SD 



 



240.0


13.4


20    -



 100  mg/kg/day


 


 Group 2



Mean 


SD 


N  %Diff 



239.6


16.7


20   


-0.2



 300  mg/kg/day


 


 Group 3



Mean 


SD 


N  %Diff 



251.2


17.1


20   


4.6



 900  mg/kg/day


 


 Group 4



Mean 


SD 


N  %Diff 



242.7


18.8


20   


1.1



 


Table 2 - Summary of Body Weights (g)


F1 Animals - Cohort 1B - Gestation



















































Sex: Female



 



 



Day(s) Relative to Mating (Litter: A)



 



0



7



14



20



 0


 mg/kg/day


 


 Group 1



Mean 


SD 



 



241.9


12.7


19    -



261.8


15.3


19    -



284.1


15.0


19    -



337.5


18.9


19    -



 100  mg/kg/day


 


 Group 2



Mean 


SD 


N  %Diff 



238.8


14.3


16   


-1.3



258.2


14.3


16   


-1.4



277.9


14.9


16   


-2.2



334.6


19.9


16   


-0.9



 300  mg/kg/day


 


 Group 3



Mean 


SD 


N  %Diff 



253.5


16.6


19   


4.8



271.6


16.9


19   


3.8



291.6


16.6


19   


2.6



348.9


19.3


19   


3.4



 900  mg/kg/day


 


 Group 4



Mean 


SD 


N  %Diff 



241.1


16.7


20   


-0.3



260.8


20.7


20   


-0.4



278.0


23.3


20   


-2.1



336.5


32.1


20   


-0.3



 


Table 2 - Summary of Body Weights (g)


F1 Animals - Cohort 1B Females - Lactation

























































































































































Group / 14Day1421
Sex7
1FMean264.2268.9276.9290.6284.2
  SD14.412.914.916.817.9
 N1817181818
 Mean262.3271.3275.8288.6286.3
2F SD14.51514.216.615.4
 N1615161616
 %Diff G1-0.70.9-0.4-0.70.8
 Mean278.4a288.5b292.6a306.2a302.1a
  SD18.916.615.715.616.2
3FN1919191919
 %Diff G15.47.35.65.46.3
 Mean265.9276.3283.6292.4291.5
4F SD21.624.520.321.722.9
 N2019202019
 %Diff G10.62.72.40.62.6

 


Table 2 - Summary of Body Weights (g)


F1 Animals - Cohort 2A






































































Sex: Male



 



 



 



 



Day(s) Relative to Start Date



 



 



 



1 [G]



2 [I]



6 [G]



13 [G]



20 [G]



27 [G]



34 [G]



 0


 mg/kg/day


 


 Group 1



Mean 


SD 



 



54.6


4.7


10    -



-


-


-


-



68.4


8.0


10    -



115.1


11.9


10    -



169.5


12.1


10    -



219.4


13.6


10    -



269.5


18.2


10    -



 100  mg/kg/day


 


 Group 2



Mean 


SD 


N  %Diff 



54.6


5.5


9   


-0.1



59.0n


7.1


2    -



67.0


8.9


9   


-2.0



111.0


12.0


10   


-3.6



160.7


12.0


10   


-5.2



205.3


14.3


10   


-6.4



250.8


18.2


10   


-6.9



 300  mg/kg/day


 


 Group 3



Mean 


SD 


N  %Diff 



58.5


5.9


10   


7.1



76.0n


-


1    -



74.7


10.9


9   


9.2



122.9


14.3


10   


6.8



177.1


16.3


10   


4.5



224.4


19.6


10   


2.3



273.9


23.0


10   


1.6



 900  mg/kg/day


 


 Group 4



Mean 


SD 


N  %Diff 



55.4


3.6


9   


1.5



59.0n 7.1


2    -



67.3


9.2


9   


-1.6



112.4


11.1


10   


-2.3



164.3


12.4


10   


-3.1



211.4


15.1


10   


-3.6



259.5


21.4


10   


-3.7



 































































Sex: Male



 



 



 



Day(s) Relative to Start Date



 



 



41



48



55



58



59



60



 0


 mg/kg/day


 


 Group 1



Mean 


SD 



 



308.0


21.9


10    -



342.0


24.0


10    -



368.4


25.7


10    -



369.8


24.4


4    -



396.8


32.0


5    -



405.0


37.8


3    -



 100  mg/kg/day


 


 Group 2



Mean 


SD 


N  %Diff 



287.9


19.6


10   


-6.5



320.4


24.2


10   


-6.3



347.9


26.8


10   


-5.6



376.8


42.4


4   


1.9



369.8


20.2


6   


-6.8



360.5


16.2


4    -11.0



 300  mg/kg/day


 


 Group 3



Mean 


SD 


N  %Diff 



313.5


26.9


10   


1.8



348.1


27.9


10   


1.8



372.8


25.9


10   


1.2



403.3


32.3


6   


9.1



404.0


54.1


3   


1.8



369.5


14.5


4   


-8.8



 900  mg/kg/day


 


 Group 4



Mean 


SD 


N  %Diff 



298.6


22.8


10   


-3.1



326.5


27.2


10   


-4.5



347.3


31.0


10   


-5.7



362.0


46.0


4   


-2.1



376.9


25.2


7   


-5.0



341.5n 0.7


2    -15.7



 






































































Sex: Female



 



 



 



 



Day(s) Relative to Start Date



 



 



 



1 [G]



2 [I]



6 [G]



13 [G]



20 [G]



27 [G]



34 [G]



 0


 mg/kg/day


 


 Group 1



Mean 


SD 



 



51.5


4.2


10    -



-


-


-


-



62.9


6.5


10    -



98.9


9.8


10    -



128.6


9.3


10    -



149.3


8.6


10    -



166.7


9.1


10    -



 100  mg/kg/day


 


 Group 2



Mean 


SD 


N  %Diff 



54.2


2.9


9   


5.3



54.0n


2.8


2    -



65.4


5.5


9   


4.0



100.5


8.6


10   


1.6



131.3


8.2


10   


2.1



155.1


8.6


10   


3.9



172.1


10.8


10   


3.2



 300  mg/kg/day


 


 Group 3



Mean 


SD 


N  %Diff 



55.7


7.4


10   


8.2



76.0n


-


1    -



68.8


10.0


9   


9.3



108.9


13.2


10   


10.1



145.1**


12.9


10   


12.8



167.6**


15.5


10   


12.3



186.0**


18.3


10   


11.6



 900  mg/kg/day


 


 Group 4



Mean 


SD 


N  %Diff 



53.6


3.7


9   


4.0



57.0n 5.7


2    -



63.2


5.3


9   


0.5



100.5


6.2


10   


1.6



135.7


10.3


10   


5.5



159.2


10.4


10   


6.6



179.3


12.7


10   


7.6



 






























































Sex: Female



 



 



Day(s) Relative to Start Date



 



 



41 [G]



48 [G]



55 [G]



58 [G]



59 [G1]



60 [G]



 0


 mg/kg/day


 


 Group 1



Mean 


SD 



 



179.8


10.1


10    -



193.9


9.9


10    -



203.7


10.7


10    -



204.8


13.1


4    -



214.0


12.7


5    -



214.3


12.2


3    -



 100  mg/kg/day


 


 Group 2



Mean 


SD 


N  %Diff 



185.7


12.3


10   


3.3



199.7


13.1


10   


3.0



210.3


14.1


10   


3.2



206.8


12.0


4   


1.0



218.0


20.1


6   


1.9



210.8


12.0


4   


-1.7



 300  mg/kg/day


 


 Group 3



Mean 


SD 


N  %Diff 



202.3**


20.4


10   


12.5



215.0**


20.3


10   


10.9



223.8*


23.4


10   


9.9



237.2


33.5


6   


15.8



247.0


47.1


3   


15.4



223.5


14.2


4   


4.3



 900  mg/kg/day


 


 Group 4



Mean 


SD 


N  %Diff 



198.6*


14.7


10   


10.5



210.7*


14.0


10   


8.7



218.6


13.4


10   


7.3



223.0


22.0


4   


8.9



229.9


10.5


7   


7.4



220.7


6.0


3   


3.0



 


 


Tables 20. Summary of Clinical Chemistry Values


F0 Animals - Males. Day: 137 Relative to Start Date






































































Sex: Male



 



 



 



 



Reporting Biochemistry



 



 



 



AST


(U/L)


[G]



ALT


(U/L)


[G]



ALP


(U/L)


[G]



GGT


(U/L)


[G1]



CK


(U/L)


[G1]



TBIL


(umol/L)


[G1]



UREA


(mmol/L)


[G]



 0


 mg/kg/day


 


 Group 1



Mean


SD


N


 



71.4  


12.3  


10      -



51.0  


11.7  


10      -



128.5  


12.4  


10      -



1.5  


0.0  


10      -



217.1   68.4  


10      -



1.25  


0.00  


10        -



7.35  


0.66  


10        -



 100  mg/kg/day


 


 Group 2



Mean


SD


N


tCtrl



74.6  


10.3  


9     


1.04



49.1  


12.0  


9     


0.96



126.0  


25.8  


9     


0.98



1.5  


0.0  


9     


1.00



292.7  


163.0  


9     


1.35



1.25  


0.00  


9       


1.00  



6.90  


0.68  


9       


0.94  



 300  mg/kg/day


 


 Group 3



Mean


SD


N


tCtrl



60.5  *


4.9  


10     


0.85



44.5  


7.8  


10     


0.87



125.1  


32.5  


10     


0.97



1.5  


0.0  


10     


1.00



213.2   70.8  


10     


0.98



1.25  


0.00  


10       


1.00  



7.16  


0.35  


10       


0.97  



 900  mg/kg/day


 


 Group 4



Mean


SD


N


tCtrl



61.6  *


5.4  


10     


0.86



36.8   14.1  


9     


0.72



115.4  


28.5  


10     


0.90



1.5   0.0  


10     


1.00



196.8   60.0  


10     


0.91



1.25   0.00  


10       


1.00  



6.94   0.68  


10       


0.94  



[G] - Anova & Dunnett: * = p ≤ 0.05
[G1] - Kruskal-Wallis & Dunn


 





































































Sex: Male



 



 



 



reporting Biochemistry



 



 



 



CREAT


(umol/L)


[G]



GLUC


(mmol/L)


[G]



CHOL


(mmol/L)


[G]



TRIG


(mmol/L)


[G]



TPROT


(g/L)


[G]



ALB


(g/L)


[G]



GLOB


(g/L)


[G]



 0


 mg/kg/day


 


 Group 1



Mean


SD


N


 



34.2  


3.2  


10      -



8.251  


0.699  


10          -



1.789  


0.249  


10          -



1.949  


0.657  


10          -



66.29  


1.89  


10        -



40.36  


1.53  


10        -



25.93  


2.18  


10        -



 100  mg/kg/day


 


 Group 2



Mean


SD


N tCtrl



32.1  


3.5  


9     


0.94



8.778  


1.195  


9         


1.06    



1.853  


0.393  


9         


1.04    



1.933  


0.681  


9         


0.99    



66.09   3.31  


9       


1.00  



41.22   2.83  


9       


1.02  



24.87   2.61  


9       


0.96  



 300  mg/kg/day


 


 Group 3



Mean


SD


N tCtrl



29.8  *


2.9  


10     


0.87



8.113  


0.521  


10         


0.98    



2.170  *


0.404  


10         


1.21    



1.594  


0.199  


10         


0.82    



66.67  


2.46  


10       


1.01  



41.93  


1.91  


10       


1.04  



24.74  


2.11  


10       


0.95  



 900  mg/kg/day


 


 Group 4



Mean


SD


N tCtrl



31.5  


3.6  


10     


0.92



8.208   0.634  


10         


0.99    



2.327  **


0.270  


10         


1.30    



1.457   0.567  


10         


0.75    



68.21  


2.50  


10       


1.03  



43.17  *


2.11  


10       


1.07  



25.04  


2.86  


10       


0.97  



 































































Sex: Male



 



 



 



Reporting Biochemistry



 



 



A/G


(ratio) [G]



CA


(mmol/L)


[G]



NA


(mmol/L)


[G]



K


(mmol/L)


[G]



CL


(mmol/L)


[G]



PHOS


(mmol/L)


[G]



 0


 mg/kg/day


 


 Group 1



Mean


SD


N


 



1.58  


0.18  


10        -



2.618  


0.059  


10          -



141.1  


1.3  


10      -



4.98  


0.29  


10        -



103.7  


1.2  


10      -



1.404  


0.193  


10          -



 100  mg/kg/day


 


 Group 2



Mean


SD


N tCtrl



1.69  


0.22  


9       


1.07  



2.637  


0.076  


9         


1.01    



141.4  


1.9  


9     


1.00



4.90  


0.34  


9       


0.98  



103.7  


1.4  


9     


1.00



1.531  


0.178  


9         


1.09    



 300  mg/kg/day


 


 Group 3



Mean


SD


N tCtrl



1.71  


0.17  


10       


1.08  



2.654  


0.055  


10         


1.01    



140.3  


2.2  


10     


0.99



4.97  


0.15  


10       


1.00  



102.3  


1.8  


10     


0.99



1.545  


0.152  


10         


1.10    



 900  mg/kg/day


 


 Group 4



Mean


SD


N tCtrl



1.75   0.28  


10       


1.11  



2.716  **


0.069  


10         


1.04    



141.3  


1.7  


10     


1.00



5.02   0.29  


10       


1.01  



101.4  **


2.0  


10     


0.98



1.591   0.187  


10         


1.13    



 


Tables 20. Summary of Clinical Chemistry Values


F0 Animals - Females. Day: 115 Relative to Start Date






































































Sex: Female



 



 



 



 



Reporting Biochemistry



 



 



 



AST


(U/L)


[G]



ALT


(U/L)


[G]



ALP


(U/L)


[G]



GGT


(U/L)


[G1]



CK


(U/L)


[G]



TBIL


(umol/L)


[G1]



UREA


(mmol/L)


[G]



 0


 mg/kg/day


 


 Group 1



Mean


SD


N


 



131.2  


28.7  


9      -



117.1  


17.5  


9      -



223.4  


49.2  


9      -



1.5  


0.0  


9      -



630.0  


384.5  


9      -



1.25  


0.00  


9        -



10.50   1.01  


9        -



 100  mg/kg/day


 


 Group 2



Mean


SD


N


tCtrl



105.3  


10.9  


8     


0.80



110.0  


18.4  


8     


0.94



200.3  


73.8  


7     


0.90



1.5  


0.0  


8     


1.00



356.8  


132.6  


8     


0.57



1.25  


0.00  


8       


1.00  



10.11   0.94  


8       


0.96  



 300  mg/kg/day


 


 Group 3



Mean


SD


N


tCtrl



116.1  


21.8  


9     


0.88



105.7  


14.1  


9     


0.90



157.1  


38.5  


9     


0.70



1.5  


0.0  


9     


1.00



614.7  


356.8  


9     


0.98



1.25  


0.00  


9       


1.00  



9.38  


0.84  


9       


0.89  



 900  mg/kg/day


 


 Group 4



Mean


SD


N


tCtrl



106.4  


25.3  


10     


0.81



100.4  


21.0  


10     


0.86



173.2  


49.5  


10     


0.78



1.5   0.0  


9     


1.00



428.3  


244.8  


9     


0.68



1.25   0.00  


9       


1.00  



10.57   1.26  


9       


1.01  



[G] - Anova & Dunnett
[G1] - Kruskal-Wallis & Dunn






































































Sex: Female



 



 



 



 



Reporting Biochemistry



 



 



 



CREAT


(umol/L)


[G]



GLUC


(mmol/L)


[G]



CHOL


(mmol/L)


[G1]



TRIG


(mmol/L)


[G]



TPROT


(g/L)


[G]



ALB


(g/L)


[G]



GLOB


(g/L)


[G]



 0


 mg/kg/day


 


 Group 1



Mean


SD


N


 



29.4  


3.2  


9      -



4.740  


1.150  


9          -



2.828  


0.449  


9          -



0.589  


0.061  


9          -



56.30   2.58  


9        -



40.21   1.79  


9        -



16.09   2.22  


9        -



 100  mg/kg/day


 


 Group 2



Mean


SD


N tCtrl



24.6  *


4.2  


8     


0.84



5.008  


0.434  


8         


1.06    



2.641  


0.493  


8         


0.93    



0.461  


0.140  


8         


0.78    



55.21   2.41  


8       


0.98  



39.51   2.62  


8       


0.98  



15.70   0.72  


8       


0.98  



 300  mg/kg/day


 


 Group 3



Mean


SD


N tCtrl



24.0  * 2.5  


8     


0.82



4.863  


0.808  


8         


1.03    



2.889  


0.655  


9         


1.02    



0.535  


0.177  


8         


0.91    



56.38   2.85  


8       


1.00  



40.59   2.45  


9       


1.01  



16.19   1.63  


8       


1.01  



 900  mg/kg/day


 


 Group 4



Mean


SD


N tCtrl



25.6  


4.6  


9     


0.87



5.003   0.815  


9         


1.06    



2.827   0.925  


9         


1.00    



0.479   0.115  


9         


0.81    



58.48   3.36  


9       


1.04  



42.81  


2.93  


10       


1.06  



15.68   1.98  


9       


0.97  



 































































Sex: Female



 



 



 



Reporting Biochemistry



 



 



A/G


(ratio) [G]



CA


(mmol/L)


[G]



NA


(mmol/L)


[G]



K


(mmol/L)


[G]



CL


(mmol/L)


[G]



PHOS


(mmol/L)


[G]



 0


 mg/kg/day


 


 Group 1



Mean


SD


N


 



2.53  


0.34  


9        -



2.680  


0.100  


9          -



140.7  


2.1  


9      -



5.12  


0.42  


9        -



99.8  


2.2  


9      -



1.708  


0.538  


9          -



 100  mg/kg/day


 


 Group 2



Mean


SD


N tCtrl



2.54  


0.23  


8       


1.00  



2.621  


0.139  


8         


0.98    



140.0  


3.5  


8     


1.00



5.05  


0.57  


8       


0.99  



100.1  


1.9  


8     


1.00



1.649  


0.518  


8         


0.97    



 300  mg/kg/day


 


 Group 3



Mean


SD


N tCtrl



2.50  


0.29  


8       


0.99  



2.694  


0.096  


9         


1.01    



140.1  


1.3  


9     


1.00



5.24  


0.62  


9       


1.02  



99.3  


1.7  


9     


1.00



1.538  


0.510  


8         


0.90    



 900  mg/kg/day


 


 Group 4



Mean


SD


N tCtrl



2.78   0.47  


9       


1.10  



2.781   0.183  


9         


1.04    



140.1  


2.3  


10     


1.00



5.19   0.42  


10       


1.01  



97.8  


2.9  


10     


0.98



1.949   0.766  


9         


1.14    



 


Tables 20. Summary of Clinical Chemistry Values


F1 Animals - Cohort 1A. Day 70 Relative to Start Date





































































Sex: Male



 



 



 



Reporting Biochemistry



 



 



 



AST


(U/L)


[G]



ALT


(U/L)


[G]



ALP


(U/L)


[G]



GGT


(U/L)


[G1]



CK


(U/L)


[G]



TBIL


(umol/L)


[G1]



UREA


(mmol/L)


[G]



 0


 mg/kg/day


 


 Group 1



Mean


SD


N


 



68.5  


6.0  


10      -



57.6  


11.4  


10      -



153.4  


23.5  


10      -



1.5  


0.0  


9      -



239.9  


42.0  


9      -



1.25  


0.00  


9        -



7.02  


0.68  


9        -



 100  mg/kg/day


 


 Group 2



Mean


SD


N


tCtrl



63.0  


7.2  


10     


0.92



52.0  


7.9  


10     


0.90



153.5  


23.8  


10     


1.00



1.5  


0.0  


10     


1.00



239.2  


101.7  


10     


1.00



1.25  


0.00  


10       


1.00  



6.32  


0.62  


10       


0.90  



 300  mg/kg/day


 


 Group 3



Mean


SD


N


tCtrl



61.3  *


5.3  


10     


0.89



49.9  


8.6  


10     


0.87



143.2  


27.9  


10     


0.93



1.5  


0.0  


10     


1.00



210.5   55.4  


10     


0.88



1.25  


0.00  


10       


1.00  



6.84  


0.59  


10       


0.97  



 900  mg/kg/day


 


 Group 4



Mean


SD


N


tCtrl



61.2  *


4.0  


10     


0.89



42.5  **


13.0  


10     


0.74



141.5  


23.9  


10     


0.92



1.5   0.0  


10     


1.00



174.9   37.4  


10     


0.73



1.25   0.00  


10       


1.00  



7.24   0.85  


10       


1.03  



[G] - Anova & Dunnett: * = p ≤ 0.05; ** = p ≤ 0.01


[G1] - Kruskal-Wallis & Dunn





































































Sex: Male



 



 



 



Reporting Biochemistry



 



 



 



CREAT


(umol/L)


[G]



GLUC


(mmol/L)


[G]



CHOL


(mmol/L)


[G]



TRIG


(mmol/L)


[G]



TPROT


(g/L)


[G]



ALB


(g/L)


[G]



GLOB


(g/L)


[G]



 0


 mg/kg/day


 


 Group 1



Mean


SD


N


 



30.8  


4.0  


9      -



9.268  


0.551  


9          -



1.723  


0.237  


9          -



2.458  


1.001  


9          -



66.48   1.47  


9        -



41.30  


1.39  


10        -



25.16   1.24  


9        -



 100  mg/kg/day


 


 Group 2



Mean


SD


N tCtrl



28.1  


3.8  


10     


0.91



8.529  **


0.254  


10         


0.92    



1.894  


0.316  


10         


1.10    



2.030  


0.674  


10         


0.83    



66.44  


1.28  


10       


1.00  



41.71  


2.18  


10       


1.01  



24.73  


2.00  


10       


0.98  



 300  mg/kg/day


 


 Group 3



Mean


SD


N tCtrl



29.8  


2.6  


10     


0.97



8.735  


0.590  


10         


0.94    



2.007  


0.325  


10         


1.16    



1.784  


0.635  


10         


0.73    



65.87  


2.39  


10       


0.99  



41.11  


2.06  


10       


1.00  



24.76  


2.03  


10       


0.98  



 900  mg/kg/day


 


 Group 4



Mean


SD


N tCtrl



32.6  


3.7  


10     


1.06



8.617  *


0.503  


10         


0.93    



2.147  *


0.325  


10         


1.25    



1.522   0.615  


10         


0.62    



65.97  


1.76  


10       


0.99  



42.49  


1.31  


10       


1.03  



23.48  


1.78  


10       


0.93  



 































































Sex: Male



 



 



 



Reporting Biochemistry



 



 



A/G


(ratio) [G]



CA


(mmol/L)


[G]



NA


(mmol/L)


[G]



K


(mmol/L)


[G]



CL


(mmol/L)


[G1]



PHOS


(mmol/L)


[G]



 0


 mg/kg/day


 


 Group 1



Mean


SD


N


 



1.64  


0.10  


9        -



2.592  


0.033  


9          -



141.0  


0.8  


10      -



5.09  


0.30  


10        -



100.5  


0.5  


10      -



1.747  


0.191  


9          -



 100  mg/kg/day


 


 Group 2



Mean


SD


N tCtrl



1.70  


0.19  


10       


1.03  



2.620  


0.064  


10         


1.01    



142.0  


1.2  


10     


1.01



5.12  


0.30  


10       


1.01  



101.7  *


0.8  


10     


1.01



2.034  **


0.122  


10         


1.16    



 300  mg/kg/day


 


 Group 3



Mean


SD


N tCtrl



1.67  


0.19  


10       


1.02  



2.626  


0.074  


10         


1.01    



141.1  


1.5  


10     


1.00



4.99  


0.20  


10       


0.98  



101.3  


1.2  


10     


1.01



1.968  **


0.179  


10         


1.13    



 900  mg/kg/day


 


 Group 4



Mean


SD


N tCtrl



1.83   0.17  


10       


1.11  



2.657   0.064  


10         


1.02    



141.6  


2.0  


10     


1.00



4.95   0.18  


10       


0.97  



99.9  


1.6  


10     


0.99



2.065  **


0.116  


10         


1.18    



 






































































Sex: Female



 



 



 



 



Reporting Biochemistry



 



 



 



AST


(U/L)


[G]



ALT


(U/L)


[G]



ALP


(U/L)


[G]



GGT


(U/L)


[G1]



CK


(U/L)


[G]



TBIL


(umol/L)


[G1]



UREA


(mmol/L)


[G]



 0


 mg/kg/day


 


 Group 1



Mean


SD


N


 



78.0  


10.7  


7      -



55.9  


14.4  


7      -



100.6  


26.1  


7      -



1.5  


0.0  


7      -



295.7  


114.0  


7      -



1.25  


0.00  


7        -



8.09  


1.15  


7        -



 100  mg/kg/day


 


 Group 2



Mean


SD


N


tCtrl



71.9  


7.5  


9     


0.92



54.6  


17.9  


9     


0.98



93.2  


23.3  


9     


0.93



1.5  


0.0  


9     


1.00



242.9  


83.5  


9     


0.82



1.25  


0.00  


9       


1.00  



7.43  


0.78  


9       


0.92  



 300  mg/kg/day


 


 Group 3



Mean


SD


N


tCtrl



68.3  


6.1  


10     


0.88



49.9  


13.2  


10     


0.89



94.3  


24.1  


10     


0.94



1.5  


0.0  


10     


1.00



259.2   98.3  


10     


0.88



1.25  


0.00  


10       


1.00  



7.70  


0.89  


10       


0.95  



 900  mg/kg/day


 


 Group 4



Mean


SD


N


tCtrl



73.7  


9.9  


10     


0.94



59.8   26.8  


10     


1.07



135.4  


38.1  


10     


1.35



2.2   1.5  


10     


1.47



296.8  


105.5  


10     


1.00



1.25   0.00  


10       


1.00  



7.66   1.24  


10       


0.95  



 






































































Sex: Female



 



 



 



 



Reporting Biochemistry



 



 



 



CREAT


(umol/L)


[G]



GLUC


(mmol/L)


[G]



CHOL


(mmol/L)


[G]



TRIG


(mmol/L)


[G]



TPROT


(g/L)


[G]



ALB


(g/L)


[G]



GLOB


(g/L)


[G]



 0


 mg/kg/day


 


 Group 1



Mean


SD


N


 



34.0  


3.5  


7      -



8.577  


0.866  


7          -



1.589  


0.267  


7          -



1.421  


0.304  


7          -



65.96   2.49  


7        -



45.56   2.60  


7        -



20.40   2.41  


7        -



 100  mg/kg/day


 


 Group 2



Mean


SD


N tCtrl



34.8  


4.5  


9     


1.02



7.801  


0.566  


9         


0.91    



1.512  


0.297  


9         


0.95    



1.021  


0.261  


9         


0.72    



67.76   2.46  


9       


1.03  



46.70   1.56  


9       


1.03  



21.06   1.80  


9       


1.03  



 300  mg/kg/day


 


 Group 3



Mean


SD


N tCtrl



32.7  


4.0  


10     


0.96



8.161  


0.464  


10         


0.95    



1.647  


0.235  


10         


1.04    



1.268  


0.529  


10         


0.89    



68.21  


2.48  


10       


1.03  



46.77  


2.62  


10       


1.03  



21.44  


2.18  


10       


1.05  



 900  mg/kg/day


 


 Group 4



Mean


SD


N tCtrl



32.4  


4.5  


10     


0.95



8.082   0.391  


10         


0.94    



1.913  *


0.234  


10         


1.20    



0.978   0.268  


10         


0.69    



67.82  


2.61  


10       


1.03  



46.97  


2.19  


10       


1.03  



20.85  


1.87  


10       


1.02  



 






























































Sex: Female



 



 



Reporting Biochemistry



 



 



A/G


(ratio) [G]



CA


(mmol/L)


[G]



NA


(mmol/L)


[G]



K


(mmol/L)


[G]



CL


(mmol/L)


[G]



PHOS


(mmol/L)


[G]



 0


 mg/kg/day


 


 Group 1



Mean


SD


N


 



2.27  


0.35  


7        -



2.574  


0.035  


7          -



140.3  


1.1  


7      -



4.43  


0.42  


7        -



103.6  


1.4  


7      -



1.227  


0.104  


7          -



 100  mg/kg/day


 


 Group 2



Mean


SD


N tCtrl



2.24  


0.21  


9       


0.99  



2.581  


0.060  


9         


1.00    



140.2  


1.2  


9     


1.00



4.78  


0.29  


9       


1.08  



102.9  


1.2  


9     


0.99



1.436  


0.206  


9         


1.17    



 300  mg/kg/day


 


 Group 3



Mean


SD


N tCtrl



2.18  


0.30  


10       


0.96  



2.591  


0.053  


10         


1.01    



140.6  


0.8  


10     


1.00



4.45  


0.20  


10       


1.00  



104.1  


1.3  


10     


1.01



1.379  


0.205  


10         


1.12    



 900  mg/kg/day


 


 Group 4



Mean


SD


N tCtrl



2.30   0.28  


10       


1.01  



2.664  **


0.039  


10         


1.03    



140.4  


1.1  


10     


1.00



4.86  *


0.27  


10       


1.10  



102.2  


0.9  


10     


0.99



1.312   0.263  


10         


1.07    



 


 


Table 19. Summary of Coagulation Values


F0 Animals – Males, Day: 137 Relative to Start Date














































Sex: Male



 



 



Reporting Coagulation



 



PT


(sec)


[G]



APTT


(sec)


[G]



FIB


(g/L)


[G]



 0


 mg/kg/day


 


 Group 1



Mean


SD


N


 



10.55  


0.20  


10        -



13.51  


1.35  


10        -



2.278  


0.177   10          .



 100  mg/kg/day


 


 Group 2



Mean


SD


N tCtrl



10.43  


0.15  


10       


0.99  



12.52  


1.72  


10       


0.93  



2.282  


0.122  


10         


1.00    



 300  mg/kg/day


 


 Group 3



Mean


SD


N tCtrl



10.30  **


0.14  


10       


0.98  



12.35  


1.14  


10       


0.91  



2.286  


0.185  


10         


1.00    



 900  mg/kg/day


 


 Group 4



Mean


SD


N tCtrl



10.26  **


0.12  


10       


0.97  



11.77  


1.51  


10       


0.87  



2.454  *


0.114  


10         


1.08    



[G] - Anova & Dunnett: * = p ≤ 0.05; ** = p ≤ 0.01


 


Table 19. Summary of Coagulation Values


F0 Animals – Females, Day: 115 Relative to Start Date














































Sex: Female



 



 



Reporting Coagulation



 



PT


(sec)


[G]



APTT


(sec)


[G]



FIB


(g/L)


[G]



 0


 mg/kg/day


 


 Group 1



Mean


SD


N


 



10.19  


0.21  


9        -



11.17  


1.22  


9        -



1.713  


0.292   9          .



 100  mg/kg/day


 


 Group 2



Mean


SD


N tCtrl



10.48  


0.34  


10       


1.03  



12.17  


1.55  


10       


1.09  



1.612  


0.157  


10         


0.94    



 300  mg/kg/day


 


 Group 3



Mean


SD


N tCtrl



10.27  


0.32  


9       


1.01  



11.82  


1.49  


9       


1.06  



1.743  


0.316  


9         


1.02    



 900  mg/kg/day


 


 Group 4



Mean


SD


N tCtrl



10.39  


0.73  


10       


1.02  



11.16  


2.64  


10       


1.00  



1.682   0.425  


10         


0.98    



[G] - Anova & Dunnett


 


Table 19. Summary of Coagulation Values


F1 Animals - Cohort 1A, Day: 70 Relative to Start Date














































Sex: Male



 



 



Reporting Coagulation



 



PT


(sec)


[G]



APTT


(sec)


[G]



FIB


(g/L)


[G]



 0


 mg/kg/day


 


 Group 1



Mean


SD


N


 



10.50  


0.14  


9        -



12.27  


1.80  


9        -



2.580  


0.172   9          .



 100  mg/kg/day


 


 Group 2



Mean


SD


N tCtrl



10.40  


0.11  


10       


0.99  



12.79  


2.10  


10       


1.04  



2.516  


0.141  


10         


0.98    



 300  mg/kg/day


 


 Group 3



Mean


SD


N tCtrl



10.34  *


0.14  


10       


0.98  



12.09  


2.30  


10       


0.99  



2.655  


0.219  


10         


1.03    



 900  mg/kg/day


 


 Group 4



Mean


SD


N tCtrl



10.33  *


0.05  


9       


0.98  



11.51  


2.25  


9       


0.94  



2.707   0.250  


9         


1.05    



[G] - Anova & Dunnett: * = p ≤ 0.05














































Sex: Female



 



 



Reporting Coagulation



 



PT


(sec)


[G]



APTT


(sec)


[G]



FIB


(g/L)


[G]



 0


 mg/kg/day


 


 Group 1



Mean


SD


N


 



10.31  


0.26  


9        -



12.21  


1.52  


8        -



2.116  


0.251   9          .



 100  mg/kg/day


 


 Group 2



Mean


SD


N tCtrl



10.17  


0.17  


9       


0.99  



12.33  


1.57  


9       


1.01  



2.128  


0.172  


9         


1.01    



 300  mg/kg/day


 


 Group 3



Mean


SD


N tCtrl



10.15  


0.14  


10       


0.98  



11.65  


0.99  


10       


0.95  



2.186  


0.194  


10         


1.03    



 900  mg/kg/day


 


 Group 4



Mean


SD


N tCtrl



10.02  **


0.17  


10       


0.97  



11.71  


1.65  


10       


0.96  



2.182   0.099  


10         


1.03    



[G] - Anova & Dunnett: ** = p ≤ 0.01


 


Table 18. Summary of Hematology Values


F0 Animals – Males Day: 137 Relative to Start Date






































































Sex: Male



 



 



 



 



Reporting Hematology



 



 



 



WBC


(10^9/L)


[G]



NEUT


(10^9/L)


[G]



LYMPH


(10^9/L)


[G1]



MONO


(10^9/L)


[G]



EOS


(10^9/L)


[G]



BASO


(10^9/L)


[G1]



LUC


(10^9/L)


[G1]



 0


 mg/kg/day


 


 Group 1



Mean


SD


N


 



4.651  


0.478  


10          -



0.961  


0.257  


10          -



3.480  


0.289  


10          -



0.065  


0.014  


10          -



0.116  


0.046  


10          -



0.008  


0.004  


10          -



0.019  


0.006  


10          -



 100  mg/kg/day


 


 Group 2



Mean


SD


N tCtrl



4.340  


0.805  


10         


0.93    



0.769  


0.202  


10         


0.80    



3.390  


0.784  


10         


0.97    



0.062  


0.018  


10         


0.95    



0.096  


0.024  


10         


0.83    



0.006  


0.005  


10         


0.75    



0.016  


0.008  


10         


0.84    



 300  mg/kg/day


 


 Group 3



Mean


SD


N tCtrl



4.469  


0.783  


10         


0.96    



0.687  *


0.169  


10         


0.71    



3.605  


0.697  


10         


1.04    



0.057  


0.021  


10         


0.88    



0.089  


0.023  


10         


0.77    



0.010  


0.000  


10         


1.25    



0.019  


0.007  


10         


1.00    



 900  mg/kg/day


 


 Group 4



Mean


SD


N tCtrl



5.192   0.960  


10         


1.12    



0.854   0.199  


10         


0.89    



4.126   0.908  


10         


1.19    



0.079   0.028  


10         


1.22    



0.092   0.040  


10         


0.79    



0.009   0.006  


10         


1.13    



0.027   0.013  


10         


1.42    



[G] - Anova & Dunnett: * = p ≤ 0.05


[G1] - Kruskal-Wallis & Dunn






































































Sex: Male



 



 



 



 



Reporting Hematology



 



 



 



RBC


(10^12/L)


[G]



HGB


(g/L)


[G]



HCT


(L/L)


[G]



MCV


(fL)


[G]



MCH


(pg)


[G]



MCHC


(g/L)


[G]



RDWG


(%)


[G]



 0


 mg/kg/day


 


 Group 1



Mean


SD


N


 



8.813  


0.449  


10          -



153.6  


4.7  


10      .



0.4464  


0.0164  


10            -



50.68  


1.37  


10        -



17.43  


0.57  


10        -



343.9  


7.0  


10      -



11.88  


0.31  


10        -



 100  mg/kg/day


 


 Group 2



Mean


SD


N tCtrl



8.630  


0.303  


10         


0.98    



150.7  


6.5  


10     


0.98



0.4441  


0.0155  


10           


0.99      



51.52  


2.26  


10       


1.02  



17.46  


1.03  


10       


1.00  



338.9  


7.5  


10     


0.99



12.00  


0.48  


10       


1.01  



 300  mg/kg/day


 


 Group 3



Mean


SD


N tCtrl



8.572  


0.317  


10         


0.97    



149.6  


5.2  


10     


0.97



0.4388  


0.0129  


10           


0.98      



51.25  


1.89  


10       


1.01  



17.47  


0.71  


10       


1.00  



341.1  


6.6  


10     


0.99



12.01  


0.52  


10       


1.01  



 900  mg/kg/day


 


 Group 4



Mean


SD


N tCtrl



8.573   0.220  


10         


0.97    



147.4  


5.4  


10     


0.96



0.4364   0.0139  


10           


0.98      



50.92  


1.39  


10       


1.00  



17.17  


0.55  


10       


0.99  



337.5  


4.5  


10     


0.98



12.16  


0.48  


10       


1.02  



 







































Sex: Male



 



Reporting Hematology



PLT


(10^9/L)


[G]



RETIC


(10^9/L)


[G]



 0


 mg/kg/day


 


 Group 1



Mean


SD


N


 



781.6  


146.2  


10      -



155.83  


19.62  


10        -



 100  mg/kg/day


 


 Group 2



Mean


SD


N tCtrl



765.3  


76.5  


10     


0.98



142.51  


41.33  


10       


0.91  



 300  mg/kg/day


 


 Group 3



Mean


SD


N tCtrl



760.4  


92.6  


10     


0.97



160.37  


18.17  


10       


1.03  



 900  mg/kg/day


 


 Group 4



Mean


SD


N tCtrl



883.4  


160.3  


10     


1.13



171.47  


21.32  


10       


1.10  



 


Table 18. Summary of Hematology Values


F0 Animals – Females Day: 115 Relative to Start Date
































































Sex: Female



 



Reporting Hematology



WBC


(10^9/L)


[G]



NEUT


(10^9/L)


[G]



LYMPH


(10^9/L)


[G]



MONO


(10^9/L)


[G]



EOS


(10^9/L)


[G]



BASO


(10^9/L)


[G]



LUC


(10^9/L)


[G]



 0


 mg/kg/day


 


 Group 1



Mean


SD


N


 



5.168  


1.072  


8          -



1.669  


0.559  


8          -



3.265  


0.790  


8          -



0.109  


0.031  


8          -



0.095  


0.038  


8          -



0.011  


0.006  


8          -



0.023  


0.009  


8          -



 100  mg/kg/day


 


 Group 2



Mean


SD


N tCtrl



5.529  


0.603  


10         


1.07    



1.737  


0.408  


10         


1.04    



3.564  


0.620  


10         


1.09    



0.113  


0.025  


10         


1.04    



0.077  


0.039  


10         


0.81    



0.012  


0.008  


10         


1.07    



0.025  


0.010  


10         


1.11    



 300  mg/kg/day


 


 Group 3



Mean


SD


N tCtrl



5.866  


1.252  


10         


1.14    



1.903  


0.424  


10         


1.14    



3.694  


0.894  


10         


1.13    



0.143  


0.060  


10         


1.31    



0.091  


0.038  


10         


0.96    



0.013  


0.007  


10         


1.16    



0.026  


0.013  


10         


1.16    



 900  mg/kg/day


 


 Group 4



Mean


SD


N tCtrl



5.570   0.802  


10         


1.08    



1.762   0.315  


10         


1.06    



3.555   0.672  


10         


1.09    



0.144   0.051  


10         


1.32    



0.073   0.013  


10         


0.77    



0.008   0.004  


10         


0.71    



0.028   0.011  


10         


1.24    



 
































































Sex: Female



 



Reporting Hematology



RBC


(10^12/L)


[G]



HGB


(g/L)


[G]



HCT


(L/L)


[G]



MCV


(fL)


[G]



MCH


(pg)


[G]



MCHC


(g/L)


[G]



RDWG


(%)


[G]



 0


 mg/kg/day


 


 Group 1



Mean


SD


N


 



7.958  


0.449  


8          -



157.3  


3.4  


8      .



0.4679  


0.0124  


8            -



58.88   2.37  


8        -



19.80   0.92  


8        -



336.4   5.2  


8      -



12.00   1.06  


8        -



 100  mg/kg/day


 


 Group 2



Mean


SD


N


tCtrl



7.678  


0.380  


10         


0.96    



153.3  


5.2  


10     


0.97



0.4515  


0.0208  


10           


0.97      



58.86  


1.56  


10       


1.00  



19.99  


0.77  


10       


1.01  



339.8  


8.3  


10     


1.01



11.84  


0.79  


10       


0.99  



 300  mg/kg/day


 


 Group 3



Mean


SD


N


tCtrl



7.466  


0.606  


9         


0.94    



152.7  


5.1  


10     


0.97



0.4468  


0.0194  


9           


0.95      



59.99   2.69  


9       


1.02  



20.48   1.21  


9       


1.03  



341.6   7.1   9     


1.02



11.82  


0.97  


10       


0.99  



 900  mg/kg/day


 


 Group 4



Mean


SD


N


tCtrl



7.528   0.566  


10         


0.95    



151.3  


8.8  


10     


0.96



0.4428   0.0316  


10           


0.95      



58.85  


1.83  


10       


1.00  



20.10  


1.07  


10       


1.02  



341.5   10.8  


10     


1.02



12.17  


1.05  


10       


1.01  



 







































Sex: Female



 



Reporting Hematology



PLT


(10^9/L)


[G]



RETIC


(10^9/L)


[G]



 0


 mg/kg/day


 


 Group 1



Mean


SD


N


 



894.6  


119.2  


8      -



177.99  


20.44  


8        -



 100  mg/kg/day


 


 Group 2



Mean


SD


N tCtrl



794.3  


132.8  


10     


0.89



178.18  


31.61  


10       


1.00  



 300  mg/kg/day


 


 Group 3



Mean


SD


N tCtrl



872.4  


121.5  


9     


0.98



161.09  


22.77  


10       


0.91  



 900  mg/kg/day


 


 Group 4



Mean


SD


N tCtrl



833.4  


177.2  


10     


0.93



173.25  


30.61  


10       


0.97  



 


Table 18. Summary of Hematology Values


F1 Animals – Cohort 1A. Day 70 Relative to Start Date






































































Sex: Male



 



 



 



 



Reporting Hematology



 



 



 



WBC


(10^9/L)


[G]



NEUT


(10^9/L)


[G]



LYMPH


(10^9/L)


[G]



MONO


(10^9/L)


[G]



EOS


(10^9/L)


[G]



BASO


(10^9/L)


[G1]



LUC


(10^9/L)


[G1]



 0


 mg/kg/day


 


 Group 1



Mean


SD


N


 



6.989  


1.038  


10          -



0.930  


0.183  


10          -



5.762  


0.940  


10          -



0.121  


0.043  


10          -



0.108  


0.036  


10          -



0.008  


0.009  


10          -



0.059  


0.018  


10          -



 100  mg/kg/day


 


 Group 2



Mean


SD


N tCtrl



7.203  


0.765  


10         


1.03    



0.970  


0.322  


10         


1.04    



5.976  


0.743  


10         


1.04    



0.099  


0.031  


10         


0.82    



0.091  


0.031  


10         


0.84    



0.011  


0.006  


10         


1.38    



0.054  


0.023  


10         


0.92    



 300  mg/kg/day


 


 Group 3



Mean


SD


N tCtrl



7.256  


0.780  


10         


1.04    



0.993  


0.193  


10         


1.07    



6.050  


0.666  


10         


1.05    



0.086  


0.030  


10         


0.71    



0.074  


0.023  


10         


0.69    



0.007  


0.005  


10         


0.88    



0.047  


0.009  


10         


0.80    



 900  mg/kg/day


 


 Group 4



Mean


SD


N tCtrl



8.090  *


0.978  


10         


1.16    



1.081   0.253  


10         


1.16    



6.736   1.071  


10         


1.17    



0.115   0.030  


10         


0.95    



0.080   0.032  


10         


0.74    



0.011   0.006  


10         


1.38    



0.064   0.023  


10         


1.08    



[G] - Anova & Dunnett: * = p ≤ 0.05


[G1] - Kruskal-Wallis & Dunn


 





































































Sex: Male



 



 



 



Reporting Hematology



 



 



 



RBC


(10^12/L)


[G]



HGB


(g/L)


[G]



HCT


(L/L)


[G]



MCV


(fL)


[G]



MCH


(pg)


[G]



MCHC


(g/L)


[G]



RDWG


(%)


[G]



 0


 mg/kg/day


 


 Group 1



Mean


SD


N


 



9.078  


0.461  


10          -



163.2  


4.9  


10      .



0.4747  


0.0207  


10            -



52.33  


1.52  


10        -



17.99  


0.90  


10        -



343.7   10.9  


10      -



11.47  


0.49  


10        -



 100  mg/kg/day


 


 Group 2



Mean


SD


N tCtrl



8.551  *


0.488  


10         


0.94    



156.9  *


5.4  


10     


0.96



0.4471  **


0.0181  


10           


0.94      



52.33  


2.09  


10       


1.00  



18.37  


0.87  


10       


1.02  



350.8  


6.2  


10     


1.02



11.59  


0.52  


10       


1.01  



 300  mg/kg/day


 


 Group 3



Mean


SD


N tCtrl



8.522  **


0.298  


10         


0.94    



157.1  *


4.1  


10     


0.96



0.4519  *


0.0171  


10           


0.95      



53.06  


1.54  


10       


1.01  



18.44  


0.54  


10       


1.03  



347.7  


7.0  


10     


1.01



11.63  


0.51  


10       


1.01  



 900  mg/kg/day


 


 Group 4



Mean


SD


N tCtrl



8.340  **


0.285  


10         


0.92    



154.7  **


4.4  


10     


0.95



0.4456  **


0.0166  


10           


0.94      



53.45  


1.51  


10       


1.02  



18.56  


0.67  


10       


1.03  



347.4  


7.6  


10     


1.01



11.84  


0.32  


10       


1.03  



[G] - Anova & Dunnett: * = p ≤ 0.05; ** = p ≤ 0.01


 







































Sex: Male



 



Reporting Hematology



PLT


(10^9/L)


[G]



RETIC


(10^9/L)


[G]



 0


 mg/kg/day


 


 Group 1



Mean


SD


N


 



763.2  


131.0  


10      -



193.85  


15.23  


10        -



 100  mg/kg/day


 


 Group 2



Mean


SD


N tCtrl



876.8  


89.5  


10     


1.15



171.56  


23.85  


10       


0.89  



 300  mg/kg/day


 


 Group 3



Mean


SD


N tCtrl



899.7  


76.1  


10     


1.18



198.49  


23.89  


10       


1.02  



 900  mg/kg/day


 


 Group 4



Mean


SD


N tCtrl



823.3  


144.5  


10     


1.08



208.78  


30.71  


10       


1.08  



 






































































Sex: Female



 



 



 



 



Reporting Hematology



 



 



 



WBC


(10^9/L)


[G]



NEUT


(10^9/L)


[G1]



LYMPH


(10^9/L)


[G]



MONO


(10^9/L)


[G1]



EOS


(10^9/L)


[G]



BASO


(10^9/L)


[G]



LUC


(10^9/L)


[G]



 0


 mg/kg/day


 


 Group 1



Mean


SD


N


 



5.104  


1.013  


9          -



0.587  


0.094  


9          -



4.321  


0.997  


9          -



0.070  


0.018  


9          -



0.081  


0.019  


9          -



0.003  


0.005  


9          -



0.038  


0.010  


9          -



 100  mg/kg/day


 


 Group 2



Mean


SD


N tCtrl



5.207  


1.111  


9         


1.02    



0.671  


0.142  


9         


1.14    



4.371  


1.058  


9         


1.01    



0.053  


0.017  


9         


0.76    



0.074  


0.028  


9         


0.92    



0.004  


0.005  


9         


1.33    



0.029  


0.014  


9         


0.76    



 300  mg/kg/day


 


 Group 3



Mean


SD


N tCtrl



4.656  


0.870  


10         


0.91    



0.708  


0.238  


10         


1.21    



3.780  


0.791  


10         


0.87    



0.060  


0.021  


10         


0.86    



0.067  


0.020  


10         


0.83    



0.003  


0.005  


10         


0.90    



0.034  


0.008  


10         


0.90    



 900  mg/kg/day


 


 Group 4



Mean


SD


N tCtrl



5.515   1.387  


10         


1.08    



0.681   0.323  


10         


1.16    



4.640   1.417  


10         


1.07    



0.076   0.039  


10         


1.09    



0.079   0.031  


10         


0.97    



0.004   0.005  


10         


1.20    



0.037   0.014  


10         


0.98    



[G] - Anova & Dunnett


[G1] - Kruskal-Wallis & Dunn


 






































































Sex: Female



 



 



 



 



Reporting Hematology



 



 



 



RBC


(10^12/L)


[G]



HGB


(g/L)


[G]



HCT


(L/L)


[G]



MCV


(fL)


[G]



MCH


(pg)


[G]



MCHC


(g/L)


[G]



RDWG


(%)


[G]



 0


 mg/kg/day


 


 Group 1



Mean


SD


N


 



7.842  


0.436  


9          -



151.6  


5.3  


9      .



0.4248  


0.0164  


9            -



54.23   1.82  


9        -



19.38   0.95  


9        -



356.8   8.1  


9      -



10.70   0.38  


9        -



 100  mg/kg/day


 


 Group 2



Mean


SD


N tCtrl



7.868  


0.225  


9         


1.00    



148.8  


6.5  


9     


0.98



0.4244  


0.0173  


9           


1.00      



53.91   1.31  


9       


0.99  



18.89   0.64  


9       


0.97  



350.7   8.5   9     


0.98



10.91   0.36  


9       


1.02  



 300  mg/kg/day


 


 Group 3



Mean


SD


N tCtrl



7.867  


0.362  


10         


1.00    



149.7  


5.4  


10     


0.99



0.4244  


0.0143  


10           


1.00      



54.01  


2.25  


10       


1.00  



19.06  


0.93  


10       


0.98  



352.6  


4.3  


10     


0.99



10.68  


0.35  


10       


1.00  



 900  mg/kg/day


 


 Group 4



Mean


SD


N tCtrl



7.744   0.368  


10         


0.99    



144.3  


5.5  


10     


0.95



0.4116   0.0156  


10           


0.97      



53.20  


1.94  


10       


0.98  



18.64  


0.75  


10       


0.96  



350.5  


6.8  


10     


0.98



10.82  


0.33  


10       


1.01  



 







































Sex: Female



 



Reporting Hematology



PLT


(10^9/L)


[G]



RETIC


(10^9/L)


[G1]



 0


 mg/kg/day


 


 Group 1



Mean


SD


N


 



833.0  


141.8  


9      -



183.07  


34.30  


9        -



 100  mg/kg/day


 


 Group 2



Mean


SD


N tCtrl



801.3  


145.2  


8     


0.96



178.02  


22.41  


9       


0.97  



 300  mg/kg/day


 


 Group 3



Mean


SD


N tCtrl



796.3  


97.2  


9     


0.96



185.18  


28.61  


10       


1.01  



 900  mg/kg/day


 


 Group 4



Mean


SD


N tCtrl



910.6  


58.3  


10     


1.09



177.80  


38.40  


10       


0.97  



 


 


Table 6. Summary of Reproductive Performance


F0 Females








































Sex: Female


Day(s) Relative to Pairing (Litter: A



)



0  mg/kg/day


 Group 1



100  mg/kg/day


 Group 2



300  mg/kg/day


 Group 3



900  mg/kg/day


 Group 4



 Group Size - Females



 


N+ve


N+ve


Mean


SD


N


%Diff


N+ve


%


N+ve %


% ProA



25     


25


25


5.3  


5.1  


25     


-


19


76.0


6


24.0


100.0


25/25



25     


25


25


3.2  


2.5  


25     


-39.4  


24


96.0


1


4.0


100.0


25/25



25     


25


25


2.7  


2.6  


25     


-49.2  


24


96.0


1


4.0


100.0


25/25



25     


24


24


3.0  


2.7  


24     


-43.2  


23


95.8


1


4.2


100.0


24/24



 Paired - Females



 Mated Females



 Pre-coital Interval (Days)



 Confirmed Mating Days 1-7



 Confirmed Mating Days 8-14



 Female Mating Index



Mated Females - Pregnant or Not Pregnant with confirmed mating date (by sperm or plug).


 


Table 6. Summary of Reproductive Performance


F1 Animals - Cohort 1B Females








































Sex: Female


Day(s) Relative to Pairing (Litter: A



)



0  mg/kg/day


 Group 1



100  mg/kg/day


 Group 2



300  mg/kg/day


 Group 3



900  mg/kg/day


 Group 4



 Group Size - Females



 


N+ve


N+ve


Mean


SD


N


%Diff


N+ve


%


N+ve %


% ProA



20     


20


20


2.7  


1.0  


20     


-


20


100.0


0


0.0


100.0


20/20



20     


20


20


3.9  


3.7  


20     


45.3  


18


90.0


2


10.0


100.0


20/20



20     


20


20


3.5  


2.7   20     


32.1  


19


95.0


1


5.0


100.0


20/20



20     


20


20


2.5  


1.2   20     


-7.5  


20


100.0


0


0.0


100.0


20/20



 Paired - Females



 Mated Females



 Pre-coital Interval (Days)



 Confirmed Mating Days 1-7



 Confirmed Mating Days 8-14



 Female Mating Index



Mated Females - Pregnant or Not Pregnant with confirmed mating date (by sperm or plug).


 


 


 


Table 7. Mating Performance and Fertility


F0 Animals




























































































































Number of Nights to  Positive Mating Sign



Group/Dose Level (mg/kg/day)



1


(0)



2


(100)



3


(300)



4


(900)



Number of Animals (Number of these not becoming pregnant)



1



6(1)*



4



8



7(1)



2



2



5



6



5



3



6



8



7



3



4



5



7



3



8



No clear indication of mating



6(1)



1



1



1



Mean number of nights to positive mating sign



2.5



2.8



2.2



2.5



Number passing one estrus



0



0



0



0



Number of males paired



25



25



25



24



Number of siring males



24



25



25



23



Male Mating Index



0.96



1.00



1.00



1.00



Male Fertility Index



1.00



1.00



1.00



0.96



Male Pregnancy Index



0.96



1.00



1.00



0.96



Female Mating Index



0.96



1.00



1.00



1.00



Female Fertility Index



1.00



1.00



1.00



0.96



Female Pregnancy Index



0.96



1.00



1.00



0.96



*Includes Animal 1524 that had 2 implant sites (no live or dead pups)


 


Table 7. Mating Performance and Fertility


F1 Animals










































































































































Number of Nights to  Positive Mating Sign



Group/Dose Level (mg/kg/day)



1


(0)



2


(100)



3


(300)



4


(900)



Number of Animals (Number of these not becoming pregnant)



1



3



5



2



7



2



5



2



4



2



3



9



5(2)



8



6



4



2



5



4



5



5



1(1)



1



0



0



6



0



0



1



0



No clear indication of mating



0



2(2)



1(1)



0



Mean number of nights to positive mating sign



2.7



2.7



2.9



2.5



Number passing one estrus



0



1



1



0



Number of males paired



20



20



20



20



Number of siring males



19



16



19



20



Male Mating Index



1.00



0.90



0.95



1.00



Male Fertility Index



0.95



0.89



0.95



1.00



Male Pregnancy Index



0.95



0.80



0.95



1.00



Female Mating Index



1.00



0.90



0.95



1.00



Female Fertility Index



0.95



0.89



0.95



1.00



Female Pregnancy Index



0.95



0.80



0.95



1.00



 


 


Table 9. Summary of Duration of Gestation and Overall Litter Performance


F0 Animals






















































































































































F0 Generation



Group/Dose Level (mg/kg/day)



 



1



2



3



4



 



(0)



(100)



(300)



(900)



Number Pregnant



24



25



25



23



Duration of Gestation (Days)


21



 


1



 


1



 


0



 


0



22



17



17



23



20



23



0



6



1



2



No clear indication of mating



6



1



1



1



Mean Duration



21.9



22.2



22.0



22.1



Number of females producing a live litter



23



25



25



23



Gestation index as %



96



100



100



100



Mean number of implant sites per pregnancy ± standard deviation



12.0 ± 3.4



12.5 ± 2.4



12.6 ± 1.9



12.9 ± 1.6



Mean total number of pups born per litter ± standard deviation



11.0 ± 3.0



11.2 ± 2.3



11.0 ± 2.5



11.0 ± 3.3



Mean number of live pups per litter ± standard deviation:


Lactation Day 1



 


11.0 ± 3.0



 


11.1 ± 2.2



 


10.9 ± 2.6



 


11.1 ± 2.6



Lactation Day 4



10.9 ± 3.1



11.2 ± 2.2



10.9 ± 2.6



11.0 ± 2.7



Lactation Day 4a



7.7 ± 1.1



7.9 ± 0.4



7.8 ± 0.7



7.7 ± 0.9



Lactation Day 7



7.7 ± 1.1



7.9 ± 0.4



7.8± 0.7



7.7 ± 0.9



Lactation Day 14



7.7 ± 1.1



7.9 ± 0.4



7.8 ± 0.7



7.7 ± 0.9



Lactation Day 21



7.7 ± 1.1



7.9 ± 0.4



7.8 ± 0.7



7.7 ± 0.9



Number of males on  Lactation Day 1 (%)



120 (47)



135 (49)



145 (53)



116 (47)



Number of females on  Lactation Day 1 (%)



134 (53)



143 (51)



127 (47)



129 (53)



a After cull of pups on PND 4 to standardise the size of the litter


 


Table 9. Summary of Duration of Gestation and Overall Litter Performance


F1 Animals















































































































































F1 Generation



Group/Dose Level (mg/kg/day)



 



1



2



3



4



 



(0)



(100)



(300)



(900)



Number Pregnant



19



16



19



20



Duration of Gestation (Days)


21



 


1



 


3



 


1



 


1



22



15



9



17



18



23



3



4



1



1



Mean Duration



22.1



22.1



22.0



22.0



Number of females producing a live litter



19



16



19



20



Gestation index as %



100



100



100



100



Mean number of implant sites per pregnancy ± standard deviation



12.6 ± 1.7



12.3 ± 2.3



13.0 ± 1.6



12.5 ± 2.3



Mean total number of pups born per litter ± standard deviation*



11.6 ± 1.9



11.1 ± 2.8



10.5 ± 2.8



11.3 ± 2.2



Mean number of live pups per litter ± standard deviation*:


Lactation Day 1



 


11.3 ± 2.2



 


11.1 ± 3.0



 


10.4 ± 2.7



 


11.1 ± 2.1



Lactation Day 4



11.3 ± 2.2



11.0 ± 2.9



10.3 ± 2.8



11.0 ± 2.1



Lactation Day 4a



7.9 ± 0.5



7.7 ± 0.9



7.6 ± 1.2



7.9 ± 0.5



Lactation Day 7



7.9 ± 0.5



7.7 ± 0.9



7.6 ± 1.2



7.9 ± 0.5



Lactation Day 14



7.9 ± 0.5



7.7 ± 0.9



7.6 ± 1.2



7.9 ± 0.5



Lactation Day 21



7.9 ± 0.5



7.7 ± 0.9



7.6 ± 1.2



7.8 ± 0.5



Number of males on  Lactation Day 1 (%)



96 (47)



96 (54)



93 (49)



100 (45)



Number of females on  Lactation Day 1 (%)



108 (53)



81 (46)



96 (51)



121 (55)



a After cull of pups on PND 4 to standardise the size of the litter


* Excludes Animal 1628 that had a total litter loss


 


 


Table 21. Summary of Urinalysis Values


F0 Animals - Males
Day: 131 Relative to Start Date











































































































































Sex: Male   Reporting Urinalysis 
VOLUMESPECIFICURINE pH
(mL)GRAVITY[G]
[G][G1] 
0 mg/kg/dayMean 4.321.01527.5
Group 1SD 2.940.00890.94
 1010            - 10        - 
   -   
 100  mg/kg/dayMean 5.641.01358
Group 2SD 2.990.00520.62
 101010
 tCtrl 1.3111.07
 300  mg/kg/dayMean 4.221.02158.15
Group 3SD 2.110.00860.63
 101010
 tCtrl 0.981.011.09
 900  mg/kg/dayMean 5.251.0300  **7.35   0.71   
Group 4SD 2.390.013310
 10100.98
 tCtrl 1.221.01 

 


Table 21. Summary of Urinalysis Values


F0 Animals - Females
Day: 108 Relative to Start Date











































































































































Sex: Female   Reporting Urinalysis 
VOLUMESPECIFICURINE pH
(mL)GRAVITY[G1]
[G][G] 
0 mg/kg/dayMean 4.851.01888.6
Group 1SD 2.330.00520.32
 10        - 10            - 10        - 
     
 100  mg/kg/dayMean 5.231.01928.45
Group 2SD 2.960.00690.16
 N101010
  tCtrl 1.0810.98
 300  mg/kg/dayMean 6.011.01998.2
Group 3SD 1.680.00390.42
 101010
 tCtrl 1.2410.95
 900  mg/kg/dayMean 7.05   2.58   1.0218   0.0051   8.05  *
Group 4SD 10100.72
 1.45110
 tCtrl   0.94

[G] - Anova & Dunnett
[G1] - Kruskal-Wallis & Dunn: * = p ≤ 0.05


 


Table 21. Summary of Urinalysis Values


F1 Animals - Cohort 1A
Day: 60 Relative to Start Date











































































































































Sex: Male   Reporting urinalysis 
VOLUMESPECIFICURINE pH
(mL)GRAVITY[G]
[G][G] 
0 mg/kg/dayMean 2.671.01828.11
Group 1SD 1.870.00780.6
 10        - 10            - 9        - 
     
 100  mg/kg/dayMean 3.071.01687.9
Group 2SD 0.950.01230.61
 101010
 tCtrl 1.1510.97
 300  mg/kg/dayMean 1.961.0302  *7.11  ** 0.70   
Group 3SD 1.250.01189
 10100.88
 tCtrl 0.731.01 
 900  mg/kg/dayMean 2.94   1.43   1.0346  **6.25  **
Group 4SD 100.00910.42
 1.11010
 tCtrl  1.020.77

 


Day: 60 Relative to Start Date


 














































Sex: Female



 



 



Reporting Urinalysi



s



VOLUME


(mL)


[G]



SPECIFIC


GRAVITY


[G]



URINE pH


[G]



 0


 mg/kg/day


 


 Group 1



Mean


SD


N


 



1.88  


1.25  


10        -



1.0226  


0.0117  


10            -



7.00  


0.94  


9        -



 100  mg/kg/day


 


 Group 2



Mean


SD


N tCtrl



3.20  


2.88  


10       


1.70  



1.0161  


0.0086  


10           


0.99      



7.15  


0.91  


10       


1.02  



 300  mg/kg/day


 


 Group 3



Mean


SD


N tCtrl



1.33  


1.08  


10       


0.71  



1.0333  


0.0118  


10           


1.01      



6.61  


0.55  


9       


0.94  



 900  mg/kg/day


 


 Group 4



Mean


SD


N tCtrl



2.61   1.55  


9       


1.39  



1.0267   0.0087  


9           


1.00      



6.44   0.68  


9       


0.92  



 


[G] - Anova & Dunnett


 


 


 


 


 


 


 


 

Applicant's summary and conclusion

Conclusions:
The administration of Omnirad 184 once daily (or twice daily 3 hours apart on some occasions) via
oral gavage was well tolerated in Han Wistar rats at dose levels of up to 900 mg/kg/day with no
test item-related deaths. There were occasional observations of decreased activity, abnormal, un
coordinated gait and low carriage for F0 animals at 900 mg/kg/day and for F0 and F1 animals, dose le
vel-related salivation and/or ploughing. F0 and F1 males at 900 mg/kg/day had slightly lower body we
ight gain, and F0 and F1 females a slightly higher dose level-related body weight gain at ≥ 100 mg/kg/
day. Food consumption was generally slightly higher for males and females at ≥ 100 mg/kg/day. Ther
e were no test item-related effects on F0 and F1 generation fertility, mating or reproduction, or F1 and
F2 generation pup numbers, survival or development. For males at 900 mg/kg/day, motor activity
was 9.6% or 12.7% lower overall for basic or fine movements, respectively, but this was not confirme
d by overall ambulation values, or any associated clinical findings, detailed functional observations or
brain morphometry values for Cohort 2A. Triglyceride values were lower for F0 males given ≥300 mg/
kg/day, F0 females and F1 animals given ≥100 mg/kg/day, and cholesterol values higher for F0 male
s only at ≥300 mg/kg/day and F1 animals at 900 mg/kg/day. There were slightly higher TSH levels
(up to 3.1-fold of controls) on the morning of necropsy in F0 and F1 adult males given 300 or 900 mg/
kg/day. There were no test item-related effects on haematology, coagulation, urinalysis, T4 values
or the immune cell populations analysed for immunophenotyping, at up to 900 mg/kg/day. Target
organ effects were observed at dose levels of ≥100 mg/kg/day and consisted of centrilobular hypertro
phy in the liver and stomach mucosal hyperplasia of the glandular region. Higher liver and/or kidney
weights were seen in both sexes. In neurological investigations, there were no test item-related brain
weight differences, gross or histological findings or effects on the brain dimensions. There were no
histological findings in the reproductive tissues of the females that had not been observed to litter tha
t would have caused reduced fertility. The NOAEL (No Observed Adverse Effect Level) for F0 and F1
adult toxicity, male and female fertility and reproduction was considered to be 900 mg/kg/day as the i
n-life and pathological findings were considered not to be adverse. For F1 and F2 developmental para
meters, the NOAEL was considered to be 900 mg/kg/day as there were no test item-related effects on
the pup numbers, survival or development.
Executive summary:

The objective of this study was to determine the potential toxicity of the chemical Omnirad 184, when given by oral gavage to adult rats and their offspring. This study was designed to provide an evaluation of reproduction and development as well as a thorough evaluation of systemic toxicity in pregnant and lactating females and their offspring. Detailed examination of key developmental endpoints, such as offspring viability, neonatal health, developmental status at birth, physical and functional development until adulthood, nervous and immune system development was expected to identify any specific target organs in the offspring. In addition, the study provided information about the effects of Omnirad 184 on the integrity and performance of the adult male and female reproductive systems and a neurobehavioural assessment of the F1 generation.


The study design was as follows:


Text Table 1 Experimental Design – F0 Animals
































































Group No.



Treatment



Dose Level (mg/kg/day)



Dose Volumea


(mL/kg)



Dose Conc.


(mg/mL)



Corrected Dose Conc.


(mg/mL)



Corrected Dose Levels (mg/kg/day)



Number of Animals



Males



Females



1



Control



0



10



0



0



0



25



25



2



Omnirad 184



100



10



10



10.03



100



25



25



3



Omnirad 184



300



10



30



30.09



301



25



25



4



Omnirad 184



900



10



90



90.27



903



25



25



a Dose volume was based on the most recent body weight measurement.
Control/vehicle = 0.5% (w/v) Carboxymethylcellulose (CMC) medium viscosity and 0.1% (w/v) Tween 80 in Milli-Q water
Conc. = concentration


Text Table 2 Experimental Design – F1 Unselected PND (Post Natal Day) 21 Pups


































Group No.a



Number of Animalsb



Males



Females



1



10



10



2



10



10



3



10



10



4



10



10



a Unselected pups were not dosed, group number refers to dam group number.


b These pups had TSH and T4 blood sampling and necropsy procedures.


 


Text Table 3 Experimental Design – Cohort 1A (Reproductive)
































































Group No.



Treatment



Dose Level (mg/kg/day)



Dose Volumea


(mL/kg)



Dose Conc.


(mg/mL)



Corrected Dose Conc.


(mg/mL)



Corrected Dose Levels (mg/kg/day)



Number of Animals



Males



Females



1



Control



0



10



0



0



0



20



20



2



Omnirad 184



100



10



10



10.03



100



20



20



3



Omnirad 184



300



10



30



30.09



301



20



21d



4



Omnirad 184



900bc



10bc



90bc



90.27



903



20



20



a Dose was based on the most recent body weight measurement.
b From 19 Aug 2021, F1 Group 4 animals were dosed twice daily (approximately 3 hours apart) at 10 mL/kg and
  450 mg/kg (45 mg/mL), equivalent to 900 mg/kg/day.


c From 30 Aug 2021, F1 Group 4 animals were dosed once daily at 10 mL/kg and 900 mg/kg/day.
d One female given 300 mg/kg/day died within minutes of receiving its first dose on Day 1, and another female
  was added to the group.


Control/vehicle = 0.5% (w/v) CMC medium viscosity and 0.1% (w/v) Tween 80 in Milli-Q water
Conc. = concentration


 


Text Table 4 Experimental Design – Surplus F1 Animals
































































Group No.



Treatment



Dose Level (mg/kg/day)



Dose Volumea


(mL/kg)



Dose Conc.


(mg/mL)



Corrected Dose Conc.


(mg/mL)



Corrected Dose Levels (mg/kg/day)



Number of Animals



Males



Females



1



Control



0



10



0



0



0



10



10



2



Omnirad 184



100



10



10



10.03



100



10



10



3



Omnirad 184



300



10



30



30.09



301



10



10



4



Omnirad 184



900bc



10bc



90bc



90.27



903



10



10



a Dose was based on the most recent body weight measurement.


b From 19 Aug 2021, F1 Group 4 animals were dosed twice daily (approximately 3 hours apart) at 10 mL/kg and 450 mg/kg (45 mg/mL), equivalent to 900 mg/kg/day.


c From 30 Aug 2021, F1 Group 4 animals were dosed once daily at 10 mL/kg and 900 mg/kg/day.
Control/vehicle = 0.5% (w/v) CMC medium viscosity and 0.1% (w/v) Tween 80 in Milli-Q water
Conc. = concentration


 


Text Table 5 Experimental Design – Cohort 1B (Reproductive)
































































Group No.



Treatment



Dose Level (mg/kg/day)



Dose Volumea


(mL/kg)



Dose Conc.


(mg/mL)



Corrected Dose Conc.


(mg/mL)



Corrected Dose Levels (mg/kg/day)



Number of Animals



Males



Females



1



Control



0



10



0



0



0



20



20



2



Omnirad 184



100



10



10



10.03



100



20



20



3



Omnirad 184



300



10



30



30.09



301



20



20



4



Omnirad 184



900bc



10bc



90bc



90.27



903



20



20



a Dose was based on the most recent body weight measurement.


b From 19 Aug 2021, F1 Group 4 animals were dosed twice daily (approximately 3 hours apart) at 10 mL/kg and 450 mg/kg (45 mg/mL), equivalent to 900 mg/kg/day.


c From 30 Aug 2021, F1 Group 4 animals were dosed once daily at 10 mL/kg and 900 mg/kg/day.
Control/vehicle = 0.5% (w/v) CMC medium viscosity and 0.1% (w/v) Tween 80 in Milli-Q water
Conc. = concentration


 


Text Table 6 Experimental Design – F2 Unselected PND 21 Pups


































Group No.a



Number of Animalsb



Males



Females



1



10



10



2



10



10



3



10



10



4



10



10



a Unselected pups were not dosed, group number refers to dam group number.


b These pups had TSH and T4 blood sampling and necropsy procedures.


 


Text Table 7 Experimental Design – Cohort 2A (Neurobehavioural)
































































Group No.



Treatment



Dose Level (mg/kg/day)



Dose Volumea


(mL/kg)



Dose Conc.


(mg/mL)



Corrected Dose Conc.


(mg/mL)



Corrected Dose Levels (mg/kg/day)



Number of Animals



Males



Females



1



Control



0



10



0



0



0



10



10



2



Omnirad 184



100



10



10



10.03



100



10



10



3



Omnirad 184



300



10



30



30.09



301



10



10



4



Omnirad 184



900bc



10bc



90bc



90.27



903



10



10



a Dose was based on the most recent body weight measurement.


b From 19 Aug 2021, F1 Group 4 animals were dosed twice daily (approximately 3 hours apart) at 10 mL/kg and 450 mg/kg (45 mg/mL), equivalent to 900 mg/kg/day.


c From 30 Aug 2021, F1 Group 4 animals were dosed once daily at 10 mL/kg and 900 mg/kg/day.
Control/vehicle = 0.5% (w/v) CMC medium viscosity and 0.1% (w/v) Tween 80 in Milli-Q water
Conc. = concentration


 


Text Table 8 Experimental Design – Cohort 2B (Neuropathology)a


































Group No.



Number of Animals



Males



Females



1



10



10



2



10



10



3



10



10



4



10



10



a Cohort 2B were not dosed as they were terminated on PND 21/22.


In Week 17 of the study (F0 dosing and the start of F1 dosing), the 900 mg/kg/day dosing formulation could not be pulled up into the gavage tube being used for newly weaned pups, due to an unknown viscosity property, resulting in some subsets of F0 and/or F1 animals either not being dosed or being given the 300 mg/kg/day formulation either once or twice daily instead (refer to section 4.6.1 for details). From 19 Aug 2021, Group 4 F1 animals were dosed twice daily 3 hours apart at 450 mg/kg (equivalent to 900 mg/kg/day) until they were old enough for the next size gavage tube on 30 Aug 2021. Although there were occasions when animals were not dosed or received a dose level lower than that required by the study protocol, there was considered to be no overall impact on the results or study outcome due to the length of time that each generation was administered the test item dose levels correctly.


The following parameters and end points were evaluated in this study: clinical observations, body weights, food consumption, estrous cycles, mating performance, fertility indices, duration of gestation and overall litter performance, litter survival indices, litter and pup weights, pre-weaning physical development of F1 pups, assessments of sexual maturation of F1 animals, clinical pathology parameters (haematology, coagulation, clinical chemistry, and urinalysis), thyroid stimulating hormone (TSH) and thyroxine (T4) analysis, gross necropsy findings, splenic immunophenotyping analysis, organ weights, sperm evaluation, ovarian follicle counts and histopathological examinations.


None of the unscheduled deaths in any generation were considered to be test item-related.


For the F0 generation at 900 mg/kg/day, test item-related clinical observations of decreased activity, abnormal/uncoordinated gait and low carriage were occasionally seen for males and/or females before pairing, and decreased activity only during lactation. For F0 and F1 generations, salivation and/or ploughing were noted for all animals given ≥300 mg/kg/day and for several animals given 100 mg/kg/day, throughout the dosing periods. F0 and F1 males given 900 mg/kg/day had slightly lower body weight gain, and F0 females a slightly higher dose level-related body weight gain at ≥ 100 mg/kg/day. Food consumption was generally slightly higher for males and females at ≥ 100 mg/kg/day.


There were no test item-related effects on F0 and F1 generation fertility, mating or reproduction, or F1 and F2 generation pup numbers, survival or development.


For males at 900 mg/kg/day, during the last 20 minutes of the 1 hour session, motor activity was 9.6% or 12.7% lower overall for basic or fine movements, respectively. Ambulation values were also lower for the same time period, but this was not confirmed by the overall ambulation values. As there were no clinical findings or detailed functional observations recorded that could have been associated with this, the cause of this apparent reduction in motor activity could not be determined.


For F0 males given ≥300 mg/kg/day and F0 females given ≥100 mg/kg/day, triglycerides were lower by up to 0.75-fold, and for males only, cholesterol higher by up to 1.3-fold. For F1 animals at ≥100 mg/kg/day, triglycerides were lower than controls by up to 0.62-fold, and at 900 mg/kg/day only, cholesterol higher by up to 1.25-fold. There were slightly higher TSH levels (up to 3.1-fold of controls) for samples taken on the morning of necropsy in F0 and F1 adult males given 300 or 900 mg/kg/day. There were no test item-related effects on haematology, coagulation, urinalysis, T4 values or the immune cell populations analysed for immunophenotyping, at up to 900 mg/kg/day.


In the F0 animals dosed from 10 weeks prior to mating and continuing throughout mating and gestation until the females were at least LD 21, microscopically, centrilobular hypertrophy in the liver was considered test item-related in all dose groups in males but only at                                  900 mg/kg/day in females and mucosal hyperplasia of the glandular region, appearing as an increased number of goblet cells, was considered test item-related at ≥ 300 mg/kg/day in males and only at 900 mg/kg/day in females. Higher liver and kidney weights were seen in both sexes at all dose levels.


In the F1 cohort 1A animals, dosed from PND 21 until at least PND 90, microscopically there was diffuse centrilobular hypertrophy in the liver. This finding was considered test item-related at all dose levels in males and only at ≥ 300 mg/kg/day in females. In the stomach, there was mucosal hyperplasia of the glandular regions, appearing as an increased number of goblet cells. This finding was considered test item-related in males and females at ≥ 300 mg/kg/day. Liver weights were higher at ≥ 300 mg/kg/day in males and females. In addition, higher kidney weights were seen in females at all dose levels and at ≥ 300 mg/kg/day in males.


In the F1 cohort 1B animals, that were used to produce an F2 generation, and that were dosed from PND 21 until at least LD 21, microscopically there was diffuse centrilobular hypertrophy in the liver. This finding was considered test item-related at all dose levels in males and only at ≥ 300 mg/kg/day in females. Liver weights were higher at all dose levels in males and only at 900 mg/kg/day in females. In addition, higher kidney weights were seen in animals dosed at 900 mg/kg/day. Stomachs were not assessed microscopically in these animals. There were no test item-related organ weight differences in the female reproductive tissues.


There were no test item-related organ weight differences in a limited list of tissues in F1 and F2 pups that were euthanised at PND 21 without active dosing.


In the F1 cohort 2A and 2B animals used for neurological investigations (euthanised on PND 21 or dosed from PND 21 until at least PND 78-80), there were no test item-related brain weight differences, gross or histological findings or effects on the brain dimensions.


There were no test item-related gross findings in the entire study, and there were only 6 premature deaths, all of which were considered not test item-related.


There were no histological findings in the reproductive tissues of the females that had not been observed to litter that would have caused reduced fertility.


In conclusion, administration of Omnirad 184 by once daily (or twice daily 3 hours apart on some occasions) oral gavage was well tolerated in Han Wistar rats at dose levels of up to 900 mg/kg/day with no test item-related deaths. There were occasional observations of decreased activity, abnormal, uncoordinated gait and low carriage for F0 animals at 900 mg/kg/day and for F0 and F1 animals, dose level-related salivation and/or ploughing. F0 and F1 males at 900 mg/kg/day had slightly lower body weight gain, and F0 and F1 females a slightly higher dose level-related body weight gain at ≥ 100 mg/kg/day. Food consumption was generally slightly higher for males and females at ≥ 100 mg/kg/day. There were no test item-related effects on F0 and F1 generation fertility, mating or reproduction, or F1 and F2 generation pup numbers, survival or development. For males at 900 mg/kg/day, motor activity was 9.6% or 12.7% lower overall for basic or fine movements, respectively, but this was not confirmed by overall ambulation values, or any associated clinical findings, detailed functional observations or brain morphometry values for Cohort 2A. Triglyceride values were lower for F0 males given ≥300 mg/kg/day, F0 females and F1 animals given ≥100 mg/kg/day, and cholesterol values higher for F0 males only at ≥300 mg/kg/day and F1 animals at 900 mg/kg/day. There were slightly higher TSH levels (up to 3.1-fold of controls) on the morning of necropsy in F0 and F1 adult males given 300 or 900 mg/kg/day. There were no test item-related effects on haematology, coagulation, urinalysis, T4 values or the immune cell populations analysed for immunophenotyping, at up to 900 mg/kg/day. Target organ effects were observed at dose levels of ≥100 mg/kg/day and consisted of centrilobular hypertrophy in the liver and stomach mucosal hyperplasia of the glandular region. Higher liver and/or kidney weights were seen in both sexes. In neurological investigations, there were no test item-related brain weight differences, gross or histological findings or effects on the brain dimensions. There were no histological findings in the reproductive tissues of the females that had not been observed to litter that would have caused reduced fertility. The NOAEL (No Observed Adverse Effect Level) for F0 and F1 adult toxicity, male and female fertility and reproduction was considered to be 900 mg/kg/day as the in-life and pathological findings were considered not to be adverse. For F1 and F2 developmental parameters, the NOAEL was considered to be 900 mg/kg/day as there were no test item-related effects on the pup numbers, survival or development.