Registration Dossier

Administrative data

Endpoint:
short-term repeated dose toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2016-10-16 to 2017-01-12
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2017
Report Date:
2017

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 422 (Combined Repeated Dose Toxicity Study with the Reproduction / Developmental Toxicity Screening Test)
Deviations:
no
Qualifier:
according to
Guideline:
other: EPA Guidelines, OPPTS 870.3650 (Combined Repeated Dose Toxicity Study with the Reproduction/Developmental Toxicity Screening Test)
Deviations:
no
Principles of method if other than guideline:
No testing guidelines were applicable for the pilot phase, as this part of the study was intended for dose level selection purposes only.
GLP compliance:
yes (incl. certificate)
Limit test:
no

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
liquid
Details on test material:
- Name of test material (as cited in study reports): JNJ-118872-AAA (T000749)
- Physical state: liquid
- Appearance: Light yellow, light brown liquid
Specific details on test material used for the study:
SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: A16EB1463
- Expiration date of the lot/batch: 2017-09-12 (retest date)
- Purity test date: Certificate of Analysis was approved on 2017-06-29

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: At room temperature
- Solubility and stability of the test substance in the solvent/vehicle: Stability of formulations for at least 3 hours at room temperature and 26 days in the refrigerator (concentration range 1-200 mg/mL) was determined as part of the analytical method development and validation study (Test Facility Study No. 512824).

FORM AS APPLIED IN THE TEST (if different from that of starting material): Light yellow solutions

OTHER SPECIFICS: correction factor is 1.04

Test animals

Species:
rat
Strain:
Wistar
Remarks:
rat
Details on species / strain selection:
This species and strain of rat has been recognized as appropriate for general and reproduction toxicity studies. Charles River Den Bosch has general and reproduction/developmental historical data in this species from the same strain and source. This animal model has been proven to be susceptible to the effects of reproductive toxicants.
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Charles River Deutschland, Sulzfeld, Germany
- Females (if applicable) nulliparous and non-pregnant: yes
- Age at study initiation: Males approx. 10-12 weeks (at start F0-treatment); Females approx. 10-12 weeks (at start pretest) and approx. 12-14 weeks (at start F0-treatment).
- Weight at study initiation: 270-309 g (males) and 205-240 g (females)
- Fasting period before study: No
- Housing:
Pretest: Females were housed in groups of 5 females/cage in Macrolon plastic cages (MIV type, height 18 cm).
Pre-mating: Animals were housed in groups of 5 animals/sex/cage in Macrolon plastic cages (MIV type, height 18 cm).
Mating: Females were caged together with males on a one-to-one-basis in Macrolon plastic cages (MIII type, height 18 cm).
Post-mating: Males were housed in their home cage (Macrolon plastic cages, MIV type, height 18 cm) with a maximum of 5 animals/cage. Females were individually housed in Macrolon plastic cages (MIII type, height 18 cm).
- Diet (e.g. ad libitum): Free access to pelleted rodent diet
- Water (e.g. ad libitum): Free access to tap-water
- Acclimation period: At least 5 days prior to start of pretest (females) or treatment (males).


ENVIRONMENTAL CONDITIONS
- Temperature (°C): 18 to 24°C
- Humidity (%): 40 to 70%
- Air changes (per hr): at least 10 room air changes/hour
- Photoperiod (hrs dark / hrs light): 12-hour light/12-hour dark cycle.

IN-LIFE DATES:
From: 2016-11-04 (start pretest, females); 2016-11-18 (start treatment); 2016-12-25/26/27/28/29 (delivery of litters)
To: 2016-12-19 (necropsy males); 2017-01-09/10/12 (necropsy females); 2017-01-08/09/11 (necropsy pups)

Administration / exposure

Route of administration:
oral: gavage
Details on route of administration:
Method: Oral gavage, using a plastic feeding tube
Frequency: Once daily for 7 days per week, approximately the same time each day with a maximum of 6 hours difference between the earliest and latest dose
Vehicle:
polyethylene glycol
Remarks:
400, specific gravity 1.125
Details on oral exposure:
PREPARATION OF DOSING SOLUTIONS:
Formulations (w/w) were prepared daily within 3 hours prior to dosing and were homogenized to a visually acceptable level. Adjustment was made for specific gravity of the vehicle and test item. A correction was made for the purity/composition of the test item. A correction factor of 1.04 was used. Formulations were placed on a magnetic stirrer during dosing.

VEHICLE
- Justification for use and choice of vehicle (if other than water): Based on trial formulations performed at Charles River Den Bosch.
- Concentration in vehicle:
From 2016-11-18 to 2016-11-23: 0 mg/mL (group 1), 11 mg/mL (group 2), 33 mg/mL (group 3), 100 mg/mL (group 4)
From 2016-11-24 to study end: 0 mg/mL (group 1), 2.2 mg/mL (group 2), 6.6 mg/mL (group 3)
- Amount of vehicle (if gavage): 1 mL/kg body weight (2016-11-18 to 2016-11-23); 5 mL/kg body weight (2016-11-24 to study end)
- Lot/batch no. (if required): no data
- Purity: no data
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
Chemical anlayses were conducted on a single occasion during the treatment phase (08 December 2016, Day 15 of treatment), according to a validated method (Test Facility Study No. 512824). One set of duplicate samples were collected and analysed. Samples of formulations were analysed for homogeneity (highest and lowest concentration) and accuracy of preparation (all concentrations). Samples of formulations were analyzed for homogeneity (highest and lowest concentration) and accuracy of preparation (all concentrations). In addition to the criteria mentioned in the validated analytical method, each calibration curve was accepted if the average of the retention times and response factors of the data points used to construct the calibration line were within a range of ±10.00% compared to those obtained during the method validation. The accuracy of preparation was considered acceptable if the mean measured concentrations were 90.00-110.00% of the target concentration. Homogeneity was demonstrated if the coefficient of variation was ≤ 10.00%. Formulations were considered stable if the relative difference before and after storage was maximally 10.00%. Stability of formulations over 3 hours at room temperature under normal laboratory light conditions (concentration range 1-200 mg/mL) was determined as part of the analytical method development and validation study (Test Facility Study No. 512824).
Duration of treatment / exposure:
6 days (Group 4); 31 days (Group 1-3 males); 52-55 days (Group 1-3 females that delivered); 41 days (female that failed to deliver).

Pups were not dosed directly but were potentially exposed to the test item in utero, via maternal milk or from exposure to maternal urine/faeces.
Frequency of treatment:
Once daily for 7 days per week, approximately the same time each day with a maximum of 6 hours difference between the earliest and latest dose.
Doses / concentrationsopen allclose all
Dose / conc.:
0 other: mg eq/kg/day
Remarks:
Group 1 (control group)
Dose / conc.:
11 other: mg eq/kg/day
Remarks:
Group 2
Dose / conc.:
33 other: mg eq/kg/day
Remarks:
Group 3
Dose / conc.:
100 other: mg eq/kg/day
Remarks:
Group 4
No. of animals per sex per dose:
10 animals/sex/dose level
Control animals:
yes, concurrent vehicle
Details on study design:
- Dose selection rationale: Dose levels were selected based on results of a dose range finding study (Test Facility Study No. 512824) in which animals were dosed for 10 days at 30, 100, 300 mg eq/kg/day. In summary, all animals in the 300 mg eq/kg group were sacrificed early on Day 6; no mortality was observed in the 100 mg eq/kg and 30 mg eq/kg groups. All animals in the 300 and 100 mg eq/kg groups displayed piloerection, hunched posture and salivation (mostly slight); lethargy was observed in all high dose animals and 1/3 mid-dose animals; ptosis was observed in all high dose animals. The only clinical sign in the 30 mg eq/kg group was slight salivation. All animals in the 300 mg eq/kg group exhibited 5-8% loss of initial body weight, while all animals at 30 and 100 mg eq/kg gained less weight than expected. Food consumption was markedly reduced in the 300 mg eq/kg group, while it was normal at 30 and 100 mg eq/kg. Upon macroscopic examination, abnormalities in forestomach were observed in 1/3 animals with many reddish foci and irregular surface; no macroscopic abnormalities were observed at 30 and 100 mg eq/kg. Liver and kidney weights were normal for all animals in all dose groups. Based on the results of this range finding study, dose levels for the main study were selected to be 0, 11, 33 and 100 mg eq/kg.

Clinical signs of toxicity at the highest dose level selected for the main study (100 mg eq/kg) generally started immediately after dosing and were still present about 3 hours after dosing (piloerection and hunched posture). Based on this finding, clinical observations (clinical signs and arena) and functional observation tests in the main study were started at least one hour (± 30 minutes) after dosing.

- Rationale for animal assignment (if not random): randomized
Positive control:
no

Examinations

Observations and examinations performed and frequency:
CAGE SIDE OBSERVATIONS: Yes
- Time schedule: At least twice daily. Animals showing pain, distress or discomfort, which was considered not transient in nature or was likely to become more severe, were sacrificed for humane reasons based on OECD guidance document on humane endpoints (ENV/JM/MONO/ 2000/7). The circumstance of any death was recorded in detail.

Note: Due to the early termination of Group 4 animals treated at 100 mg eq/kg, they were not subjected to any of the following investigations (except for body weight, post-mortem examination and limited estrous cycle evaluation).

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: Daily from start of treatment onwards up to the day prior to necropsy, detailed clinical observations were made for all animals and were started at least at 1 hour (± 30 min) after dosing. Once prior to start of treatment and at weekly intervals during the treatment period this was also performed outside the home cage in a standard arena.

BODY WEIGHT: Yes
- Time schedule for examinations: Males and females were weighed on the first day of treatment (prior to first dosing) and weekly thereafter. Mated females were weighed on Days 0, 4, 7, 11, 14, 17 and 20 post-coitum and during lactation on PND 1, 4, 7 and 13. Body weight gain was calculated and reported.

FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study):
- Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: Yes
Weekly, except for males and females which were housed together for mating and for females without evidence of mating. Food consumption of mated females was measured on Days 0, 4, 7, 11, 14, 17 and 20 post-coitum and during lactation on PND 1, 4, 7 and 13.

WATER CONSUMPTION AND COMPOUND INTAKE (if drinking water study): No
- Time schedule for examinations: Subjective appraisal was maintained during the study, but no quantitative investigation was introduced as no treatment related effect was suspected.

HAEMATOLOGY: Yes
- Time schedule for collection of blood: Blood samples were collected at the end of the treatment period on the day of scheduled necropsy from the selected 5 animals/sex/group under anaesthesia using isoflurane between 7.00 and 10.30 a.m. The animals were deprived of food overnight (with a maximum of 24 hours) before blood sampling, but water was available. Blood samples were drawn from the retroorbital sinus and collected into tubes with K3-EDTA for hematology parameters and with citrate for clotting tests.
- Parameters: white blood cells, differential leukocyte counts, red blood cells, reticulocytes, red blood cell distribution width, haemoglobin, haematocrit, mean corpuscular volume, mean corpuscular haemoglobin, mean corpuscular haemoglobin concentration, platelets, prothrombin time, activated partial thromboplastin time.

CLINICAL CHEMISTRY: Yes
- Blood samples were collected at the end of the treatment period on the day of the scheduled necropsy from the selected 5 animals/sex/group under anesthesia using isoflurane between 7.00 and 10.30 a.m. The animals were deprived of food overnight (with a maximum of 24 hours) before blood sampling, but water was available. Blood samples were drawn from the retro-orbital sinus and collected into tubes with Li-heparin for clinical biochemistry parameters. An additional blood sample was collected into serum tubes for determination of bile acids.
- Parameters: alanine aminotransferase, aspartate aminotransferase, alkaline phosphatase, total protein, albumin, total bilirubin, bile acids, urea, creatinine, glucose, cholesterol, sodium, potassium, chloride, calcium, inorganic phosphate
- Thyroid hormone analysis


FUNCTIONAL OBSERVATIONS:
- Time schedule: These tests were performed after observation for clinical signs (incl. arena observation, if applicable) on the selected 5 animals/sex/group. Selected males were tested during week 4 of treatment and the selected females were tested once during the last week of lactation.
- Parameters: hearing ability, pupillary reflex, static righting reflex, fore- and hindlimb grip strength recorded as the mean of three measurements per animal, locomotor activity







Sacrifice and pathology:
SACRIFICE
- Male animals: All surviving animals, following completion of the mating period (a minimum of 28 days of dose administration).
- Maternal animals: All surviving animals, on PND 14-16 (females which delivered) and on day 26 (one female which failed to deliver, with evidence of mating).

GROSS PATHOLOGY: Yes
- All animals surviving to the end of the observation period were deeply anaesthetized using isoflurane and subsequently exsanguinated. After sacrifice, all animals were subjected to a full post mortem examination, with special attention being paid to the reproductive organs. Descriptions of all macroscopic abnormalities were recorded.
- Necropsy was conducted as soon as possible after spontaneous death and always within 24 hours.
- Samples of the following tissues and organs of the selected 5 animals/sex/group were collected and fixed in 10% buffered formalin (neutral phosphate buffered 4% formaldehyde solution): Adrenal glands (M/F), (Aorta) (M/F), Brain - cerebellum, mid-brain, cortex (7 levels) (M/F), Caecum (M/F), Cervix (F), Clitoral gland (F), Colon (M/F), Coagulation gland (M), (Cowper’s gland) (M), Duodenum (M/F), Epididymides (M), Eyes (with optic nerve (if detectable) and Harderian gland) (M/F), Mammary gland area (M/F), Femur including joint (M/F), (Glans penis) (M), (Levator ani plus bulbocavernosus muscle complex (LABC)) (M), Heart (M/F), Ileum (M/F), Jejunum (M/F), Kidneys (M/F), (Lacrimal gland, exorbital) (M/F), (Larynx) (M/F), Liver (M/F), Lung, infused with formalin (M/F), Lymph nodes mandibular, mesenteric (M/F), (Nasopharynx) (M/F), (Esophagus) (M/F), Ovaries (F), (Pancreas) (M/F), Peyer's patches [jejunum, ileum] if detectable (M/F), Pituitary gland (M/F), Preputial gland (M), Prostate gland (M), Rectum (M/F), (Salivary glands - mandibular, sublingual) (M/F), Sciatic nerve ( M/F), Seminal vesicles (M), Skeletal muscle (M/F), (Skin) (M/F), Spinal cord -cervical, mid thoracic, lumbar (M/F), Spleen (M/F), Sternum with bone marrow (M/F), Stomach (M/F), Testes (M), Thymus(M/F), Thyroid including parathyroid if detectable (M/F), (Tongue) (M/F), Trachea (M/F), Urinary bladder (M/F), Uterus (F), Vagina (F), All gross lesions (M/F). Tissues/organs mentioned in parentheses were not examined by the pathologist, since no signs of toxicity were noted at macroscopic examination.
- Samples of the following tissues and organs of all remaining animals, males that fail to sire and females which fail to deliver, were collected and fixed in 10% buffered formalin: Cervix (F), Clitoral gland (F), Coagulation gland (M), Cowper’s glands (M), Epididymides (M), Glans penis (M), Levator ani plus bulbocavernosus muscle complex (LABC) (M), Mammary gland area (M/F), Ovaries (F), Preputial gland (M), Prostate gland (M), Seminal vesicles (M), Testes (M), Thyroid including parathyroid if detectable (M/F), Uterus (F), Vagina (F), All gross lesions (M/F).


HISTOPATHOLOGY: Yes
- All organ and tissue samples were processed, embedded and cut at a thickness of 2-4 micrometers . These slides were stained with haematoxylin and eosin. The additional slides of the testes (to examine staging of spermatogenesis) were stained with PAS/haematoxylin.
- The following slides were examined by a pathologist: The preserved organs and tissues of the selected 5 animals/sex of Groups 1 and 3 and all animals of Group 4; Additional slides of the testes of the selected 5 males of Groups 1 and 3, all males of Group 4, and male no. 28 (Group 3) that failed to sire to examine staging of spermatogenesis; All gross lesions of all animals (all dose groups); Stomach and esophagus of all selected 5 animals of Group 2 (males and females), based on treatment-related changes in this organ in Groups 3 and 4; The reproductive organs of male no.28 that failed to sire and female no.68 that failed to deliver healthy pups.
- All abnormalities were described and included in the report. An attempt was made to correlate gross observations with microscopic findings.
- A peer review on the histopathology data was performed by a second pathologist.

Other examinations:
ORGAN WEIGHTS
- Absolute organ weights and organ to body weight ratios were reported.
- The following organ weights and terminal body weight were recorded from the selected 5 animals/sex/group (Group 1-3 animals) on the scheduled day of necropsy: Adrenal glands, Brain, Epididymides, Heart, Kidneys, Liver, Ovaries, Prostate, Seminal vesicles including coagulating glands, Spleen, Testes, Thymus, Thyroid, Uterus (including cervix).
- The following organ weights and terminal body weight were recorded from all remaining animals on the scheduled day of necropsy: Epididymides, Testes, Thyroid.
Statistics:
The following statistical methods were used to analyse the data:
- If the variables could be assumed to follow a normal distribution, the Dunnett-test based on a pooled variance estimate was applied for the comparison of the treated groups and the control groups for each sex.
- The Steel-test was applied if the data could not be assumed to follow a normal distribution.
- The Fisher Exact-test was applied to frequency data.
- The Kruskal-Wallis nonparametric ANOVA test was applied to motor activity data to determine intergroup differences. All tests were two-sided and in all cases p < 0.05 was accepted as the lowest level of significance.
Group means were calculated for continuous data and medians were calculated for discrete data (scores) in the summary tables. Test statistics were calculated on the basis of exact values for means and pooled variances. Individual values, means and standard deviations may have been rounded off before printing. Therefore, two groups may display the same printed means for a given parameter, yet display different test statistics values.

Results and discussion

Results of examinations

Clinical signs:
effects observed, treatment-related
Description (incidence and severity):
Treatment-related clinical signs of toxicity were noted at 100 mg eq/kg in both sexes. Findings in females at 100 mg eq/kg included hunched posture, piloerection and a lean appearance in up to all females, starting on Day 4, slight lethargy in half of the females at Day 6, and rales in a few females starting on Day 2. Males at 100 mg eq/kg were less severely affected. They showed hunched posture from Day 5 and one male had a lean appearance.

At 33 mg eq/kg, one female showed hunched posture and piloerection between Days 5-16, and diarrhoea on Days 8-10.

Salivation was noted after dosing at all dose levels with a dose-related trend in frequency and time of onset. This salivation was considered to be a physiological response rather than a sign of systemic toxicity considering its severity and may be related to the irritancy of the test item.

No additional clinical signs of toxicity were noted during the weekly arena observations.

The only other findings were alopecia in two females (treated at 11 or 33 mg eq/kg) and single occasions of piloerection in a single female at 11 mg eq/kg and among a few females at 33 mg eq/kg. These findings were considered to be incidental and not related to treatment.
Mortality:
mortality observed, treatment-related
Description (incidence):
One female treated at 100 mg eq/kg was found dead on Day 7. Prior to death this animal showed clinical signs of toxicity including piloerection, hunched posture, lethargy, rales and a lean appearance. Similar signs of toxicity were noted in the surviving animals of this group, particularly in females. Moreover, these animals showed significant body weight loss (on average 10% or 15% of the initial weight for males and females, respectively). Therefore, all remaining animals treated at 100 mg eq/kg were sacrificed for humane reasons on the same day.

Erosion/ulceration in the esophagus and/or forestomach were considered to be the main contributors responsible for the moribund condition or death of these rats. Main test item related macroscopic findings were present in the esophagus in the form of thickened wall and grey white contents (microscopic correlates erosion/ulceration squamous epithelium, hypertrophy squamous epithelium and/or lymphogranulocytic cell infiltrate), in the forestomach in the form of irregular surface (microscopic correlate hyperplasia squamous cell) and distended with gas and mucous/gelatinous/foamy contents (no microscopic correlate), and in various parts of the intestines in the form of distended with gas and gelatinous/foamy contents (no microscopic correlate). The macroscopic and microscopic changes in the esophagus and forestomach indicated irritancy of the test item.

In addition, one control male was sacrificed in extremis on Day 19 when it showed piloerection, a pale appearance, red discolouration of the penis, and a hardened abdomen with a nodule. The large malignant nephroblastoma in the right kidney was regarded as the main cause of moribundity.
Body weight and weight changes:
effects observed, treatment-related
Description (incidence and severity):
Males and females treated at 100 mg eq/kg showed significant weight loss between Days 1-7 (on average 10 and 15%, respectively). Reduced mean body weight gain was observed in females at 33 mg eq/kg over the premating period, achieving levels of statistical significance on Day 8 and at start of the mating period when compared to controls. However, the reduction was mainly the results of marked body weight loss in one female. From start of mating onwards, body weight gain at 33 mg eq/kg was normal and body weights of these females were within the same range as controls.
Food consumption and compound intake (if feeding study):
no effects observed
Description (incidence and severity):
No treatment-related changes in food consumption before or after allowance for body weight were noted for males and females treated at 11 and 33 mg eq/kg in comparison with controls.

No data for food consumption of Group 4 are available, because these animals were sacrificed within one week after start of treatment.
Food efficiency:
not examined
Water consumption and compound intake (if drinking water study):
not examined
Ophthalmological findings:
not examined
Haematological findings:
effects observed, treatment-related
Description (incidence and severity):
Statistically significant differences in haematological parameters were limited to a lower percentage of neutrophils at 33 mg eq/kg in males. The mean value at 33 mg eq/kg was 35% lower than the concurrent control value and slightly below the historical control range. A concurrent increase in lymphocytes was observed in these males, not reaching a level of statistical significance but slightly above the historical control range.

Animals treated at 100 mg eq/kg were not subjected to haematology examinations due to their early sacrifice.
Clinical biochemistry findings:
effects observed, non-treatment-related
Description (incidence and severity):
No toxicologically relevant changes occurred in clinical biochemistry parameters of rats treated up to 33 mg eq/kg. Isolated statistically significant variations noted in clinical biochemistry parameters were unrelated to treatment or not toxicologically relevant due to the lack of a dose-related response and/or slight magnitude of the difference from controls (values in treated rats remained within normal limits).

Thyroid hormone analyses: Serum levels of T4 in F0 males were not affected by treatment.

Animals treated at 100 mg eq/kg were not subjected to clinical biochemistry examinations due to their early sacrifice.
Urinalysis findings:
not examined
Behaviour (functional findings):
effects observed, non-treatment-related
Description (incidence and severity):
Hearing ability, pupillary reflex and static righting reflex were normal in all examined animals. Grip strength was not affected by treatment.
The variation in motor activity did not indicate a relation with treatment. All groups showed a similar habituation profile with a decreasing trend in activity over the duration of the test period.

Animals treated at 100 mg eq/kg were not subjected to functional observations due to their early sacrifice.
Immunological findings:
not examined
Organ weight findings including organ / body weight ratios:
no effects observed
Description (incidence and severity):
There were no treatment-related alterations in organ weights up to 33 mg eq/kg. Organs of animals treated at 100 mg eq/kg were not weighed due to the early sacrifice of these animals.
Gross pathological findings:
effects observed, treatment-related
Description (incidence and severity):
Animals treated at 100 mg eq/kg (found dead or sacrificed moribund on Day 7): Main test item-related macroscopic findings were present in the esophagus (thickened wall and grey white contents), stomach (irregular surface of the forestomach and distended with gas and mucous/gelatinous/foamy contents) and various parts of the intestines (distended with gas and gelatinous/foamy contents).

Controls and animals treated at 11 or 33 mg eq/kg : There were no treatment-related gross observations up to 33 mg eq/kg. All of the recorded macroscopic findings were within the range of background gross observations encountered in rats of this age and strain. These necropsy findings were therefore considered to be unrelated to treatment. In control male no.10 that was sacrificed in extremis on day 19, an enlarged right kidney and an enlarged spleen were observed at necropsy. Histopathology revealed a large malignant nephroblastoma in the right kidney . This type of finding in a control animal occurred spontaneously and was considered non-study related, incidental finding.
Neuropathological findings:
not examined
Histopathological findings: non-neoplastic:
effects observed, treatment-related
Description (incidence and severity):
The test item-related findings in the stomach of males and females treated at 100 mg eq/kg consisted of lymphogranulocytic inflammation forestomach, hyperplasia squamous cell and ulcer forestomach, all up to moderate degree. The test item-related findings in the esophagus of males and females treated at 100 mg eq/kg consisted of erosion/ulceration squamous epithelium (up to massive degree), hypertrophy squamous epithelium (up to moderate degree) and lymphogranulocytic inflammatory cell infiltrate (up to marked degree).

Controls and animals treated at 11 or 33 mg eq/kg: There was a test item-related finding in the stomach of females treated at 33 mg eq/kg, consisting of hyperplasia squamous cell at minimal degree. There were no other test item-related histologic changes. The remainder of the recorded microscopic findings were within the range of background pathology encountered in rats of this age and strain. There was no test item-related alteration in the prevalence, severity, or histologic character of those incidental tissue alterations.
Histopathological findings: neoplastic:
not examined
Other effects:
not examined
Details on results:
- Analysis of dose preparations: The concentrations analysed in the formulations of Group 2, Group 3 and Group 4 were considered to be in agreement with target concentrations (i.e. mean accuracies between 90.00% and 110.00%). No test item was detected in the Group 1 formulation. The formulations of Group 2 and Group 4 were homogeneous (i.e. coefficient of variation ≤ 10.00%).

Effect levels

Key result
Dose descriptor:
NOAEL
Effect level:
33 other: mg eq/kg/day in a dose volume of 5 ml/kg
Based on:
test mat.
Sex:
male/female
Basis for effect level:
gross pathology
histopathology: non-neoplastic
other:
Remarks on result:
other: :
Remarks:
The animals had macroscopic and microscopic changes in the esophagus and stomach indicative of local toxicity resulting from the irritant properties of the test item (which is classified as a skin irritant). Main macroscopic findings in the esophagus included a thickened wall and grey white contents which correlated microscopically with erosion/ulceration of the squamous epithelium, squamous hyperplasia and/or lymphogranulocytic cell infiltrate. Gastric findings included an irregular surface of the forestomach at necropsy and histopathological changes consisting of lymphogranulocytic inflammation, squamous cell hyperplasia and/or ulceration in the forestomach. Additional macroscopic findings included distension of the stomach (with gas and mucous/gelatinous/ foamy contents) and various parts of the intestines (with gas and gelatinous/foamy contents). The morbidity of the animals treated at 100 mg eq/kg was considered to have resulted from the local toxic effects in the esophagus and stomach.

Target system / organ toxicity

Key result
Critical effects observed:
yes
Lowest effective dose / conc.:
100 other: mg eq/kg/day
System:
gastrointestinal tract
Organ:
oesophagus
stomach
Treatment related:
yes
Dose response relationship:
not specified
Relevant for humans:
not specified

Applicant's summary and conclusion

Conclusions:
In conclusion, treatment with JNJ-118872-AAA (T000749) by oral gavage in male and female Wistar Han rats at dose levels of 11, 33 and 100 mg eq/kg/day resulted in macroscopic and microscopic changes in the esophagus and stomach in the 100 mg eq/kg/day animals indicative of local toxicity resulting from the irritant properties of the test item. Based on these results, the No Observed Adverse Effect Level (NOAEL) was concluded to be 33 mg eq/kg/day in a dose volume of 5 ml/kg.

Therefore, the substance is classified for specific target organ toxicity after repeated exposure (STOT RE) category 2 according to the CLP Regulation.