Registration Dossier

Administrative data

Description of key information

Oral: 90 day repeated dose studies: 
NOAEL dog: systemic: 30 mg/kg bw/day, local: 100 mg/kg bw/d
NOAEL rat: systemic: 150 mg/kg bw/day, local: 15 mg/kg bw/d

Key value for chemical safety assessment

Repeated dose toxicity: via oral route - systemic effects

Link to relevant study records
Reference
Endpoint:
sub-chronic toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Study period:
1990-07-06 - 1990-11-02
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: GLP guideline study with acceptable restrictions. Analytical purity of test material not specified.
Qualifier:
according to
Guideline:
OECD Guideline 409 (Repeated Dose 90-Day Oral Toxicity in Non-Rodents)
Deviations:
yes
Remarks:
analytical purity of test material not specified
GLP compliance:
yes
Species:
dog
Strain:
Beagle
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Hazleton Research Products, Inc., Kalamazoo, Michigan, on July 18, 1989.
- Age at study initiation: 5-6 months
- Weight at study initiation: 8.7-10.2 kg for males, 6.6-9.3 kg for females
- Housing: Individually in elevated, stainless-steel cages
- Diet: Ad libitum
- Water: Ad libitum
- Acclimation period: 2 weeks


ENVIRONMENTAL CONDITIONS
- Temperature (°C): 16.7-25.6
- Humidity (%): 13-100%
- Air changes (per hr): 10/h
- Photoperiod (hrs dark / hrs light): 12/12
Route of administration:
oral: capsule
Vehicle:
other: Capsule
Details on oral exposure:
VEHICLE
- Concentration in vehicle: 100%
- Amount of vehicle (if gavage): 1 capsule/day
- Purity: 100%
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
Routine concentration analyses indicated that animals were generally dosed with formulations that were within ± 10% of the target range. In cases where the percent target was not within this range, formulations were remixed and reanalyzed during the study week.
Duration of treatment / exposure:
13 weeks
Frequency of treatment:
Daily, 7 days/week
Remarks:
Doses / Concentrations:
15, 30 and 100 mg/kg bw/day
Basis:
other: Nominal in capsule
No. of animals per sex per dose:
4
Control animals:
yes, concurrent vehicle
Observations and examinations performed and frequency:
CAGE SIDE OBSERVATIONS: Yes
- Time schedule: Once each day

DETAILED CLINICAL OBSERVATIONS: Yes,
- Time schedule: Twice each day

BODY WEIGHT: Yes
- Time schedule for examinations: Prior to treatment and weekly thereafter

FOOD CONSUMPTION
- Time schedule for examinations: Weekly

OPHTHALMOSCOPIC EXAMINATION: Yes
- Time schedule for examinations: On all dogs prior to treatment and on the control and high-dose dogs prior to termination of treatment

HAEMATOLOGY: Yes, cell morphology hemoglobin, corrected leukocyte count, leukocyte count, erythrocyte count, leukocyte differential, hematocrit, platelet count
- Time schedule for collection of blood: Prior to termination of dosing
- Animals fasted: Yes, overnight

CLINICAL CHEMISTRY: Yes, alanine aminotransferase, glucose, albumin, inorganic phosghorus, aspartate aminotransferase, potassium, calcium, sodium, chloride, total bilirubin, creatinine, total protein, ganrna glutamyltransferase, urea nitrogen
- Time schedule for collection of blood: Prior to termination of dosing
- Animals fasted: Yes, overnight

URINALYSIS: No

NEUROBEHAVIOURAL EXAMINATION: No

OTHER:
-Organ weights of adrenals, testes with epididymides, kidneys, thyroid (with parathyroids), liver (with gallbladder drained)
Sacrifice and pathology:
GROSS PATHOLOGY: external surface of the body, all orifices, cranial cavity, carcass, external surfaces of the brain and spinal cord and the cut surfaces of the spinal cord (at necropsy); the cut surfaces of the brain were examined at the time of tissue trimming nasal cavity and nasal turbinates, thoracic, abdominal and pelvic cavities and their viscera, cervical tissues and organs

HISTOPATHOLOGY: adrenals, aorta, brain with brainstem, (medulla/pons, cerebellar cortex, and cerebral cortex), cecum, colon, rectum, cervical spinal cord, duodenum, jejunum, iIeum, esophagus, eyes, gallbladder, heart, kidneys, lesions, liver, lumbar spinal cord, lung (with mainstem bronchi), mammary gland, mesenteric lymph node
mid-thoracic spinal cord, ovaries, pancreas, pituitary, prostate, salivary glands (mandibular), sciatic nerve wi h skeletal muscle, spleen,sternum with bone marrow, stomach, testes with epididymides, thymus, thyroid (parathyroids), trachea, urinary bladder, uterus with vagina and cervix
Other examinations:
Detailed physical examinations performed weekly
Statistics:
Group comparisons were analyzed at the 5% two-tailed probability level.
Clinical signs:
effects observed, treatment-related
Description (incidence and severity):
no adverse effects
Mortality:
mortality observed, treatment-related
Description (incidence):
no adverse effects
Body weight and weight changes:
no effects observed
Food consumption and compound intake (if feeding study):
no effects observed
Food efficiency:
not examined
Water consumption and compound intake (if drinking water study):
not examined
Ophthalmological findings:
no effects observed
Haematological findings:
no effects observed
Clinical biochemistry findings:
no effects observed
Urinalysis findings:
not examined
Behaviour (functional findings):
not examined
Organ weight findings including organ / body weight ratios:
no effects observed
Gross pathological findings:
no effects observed
Histopathological findings: non-neoplastic:
effects observed, treatment-related
Description (incidence and severity):
no adverese effects
Histopathological findings: neoplastic:
not examined
Details on results:
CLINICAL SIGNS AND MORTALITY
All animals survived until terminal sacrifice. Apparent treatment-related changes noted in the 100mg/kg bw/day group males and/or females consisted of an increased incidence of salivation and emesis. In addition, soft feces, noted with mucus alone or mucus and bile-like material, were noted with a greater incidence in the high-dose animals.

BODY WEIGHT AND WEIGHT GAIN
Statistical evaluation of mean absolute body weights and mean body weight gain values failed to reveal any significant differences when treatment groups were compared to respective control values.

FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study)
Statistical evaluation of mean total food consumption values failed to reveal any significant differences when treatment groups were compared to respective control values.

OPHTHALMOSCOPIC EXAMINATION
There were no apparent compound-related ophthalmoscopic changes observed at termination.

HAEMATOLOGY
Statistical evaluation of clinical hematology values revealed a significantly elevated mean erythrocyte value in the 100 mg/kg bw/day group females.
Although the statistical significance was noted, the 100 mg/kg bw/day group female mean value is within the historical control range; therefore, the significance appears to be associated with an unusually low mean control value. The toxicological relevance of this finding is questionable.

CLINICAL CHEMISTRY
Statistical evaluation of mean blood chemistry values revealed an elevated alanine aminotransferase value in the 100 mg/kg bw/day group females, elevated mean calcium values in the 30 and 100 mg/kg bw/day group females, and a depressed mean blood urea nitrogen value in the 30mg/kg bw/day group males. In regard to the calcium and blood urea nitrogen values, the relatively low magnitude of change in the calcium value and the absence of a dose response for the urea nitrogen value also lend question as to their biological significance.

ORGAN WEIGHTS
Statistical evaluation of organ weight values failed to reveal any significant differences when the absolute or relative organ weights were compared to respective control values.

GROSS PATHOLOGY
There were no gross necropsy findings which were considered to be related to treatment with ATMER 163. Those observations which were noted were of the type commonly observed in this species of dog at this laboratory.

HISTOPATHOLOGY: NON-NEOPLASTIC
Histomorphologic examination of protocol-specified tissues revealed an increased incidence of pigment accumulation in the Kupffer cells and bile canaliculi in the livers of the Group 4 females. There were no other apparent compound-related tissue changes noted.

Dose descriptor:
NOAEL
Remarks:
systemic
Effect level:
30 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
female
Basis for effect level:
other: tissue alterations in the liver of females at 100 mg/kg bw/d
Dose descriptor:
NOAEL
Remarks:
local
Effect level:
100 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: no adverse effects up to the limit dose of 100 mg/kg bw/d
Critical effects observed:
not specified

Based on the data generated from this study, the no-observable-adverse-effect level NOAEL of Atmer 163 when administered via capsule for approximately 13 weeks to male and female beagle dogs is 100 mg/kg bw/day for local effects and 30 mg/kg bw/d for systemic effects.

Conclusions:
CLP: not classified
Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
NOAEL
30 mg/kg bw/day
Study duration:
subchronic
Species:
rat
Quality of whole database:
The available information comprises adequate, reliable (Klimisch score 2) and consistent studies, and is thus sufficient to fulfil the standard information requirements set out in Annex VIII-IX, 8.6, of Regulation (EC) No 1907/2006.

Repeated dose toxicity: inhalation - systemic effects

Endpoint conclusion
Endpoint conclusion:
no study available

Repeated dose toxicity: inhalation - local effects

Endpoint conclusion
Endpoint conclusion:
no study available

Repeated dose toxicity: dermal - systemic effects

Endpoint conclusion
Endpoint conclusion:
no study available

Repeated dose toxicity: dermal - local effects

Endpoint conclusion
Endpoint conclusion:
no study available

Additional information

The test material, Atmer 163, was administered orally via gavage to rats for approximately 13 weeks to assess subchronic toxicity (Hazleton 564/162). Twenty rats/sex (Group 1) received deionized water and served as the control. Twenty additional rats/sex/group received the test material at levels of 15, 30, or 150 mg/kg bw/day (Groups 2, 3, and 4, respectively). The dose volume for each group was 5 mL/kg bw/day.

Parameters evaluated in the determination of toxicity were as follows: clinical signs assessed by mortality and moribundity checks performed twice each day, cageside observations performed once each day, and hands-on physical examinations performed weekly; body weights recorded prior to treatment and weekly thereafter; food consumption recorded weekly; ophthalmoscopic examinations performed prior to treatment and during Week 13; predetermined clinical pathology parameters; gross pathology; organ weight data; and microscopic examination of tissues.

Treatment-related effects were noted at the 30 and 150 mg/kg/day dose levels.

Two 30 mg/kg bw/day animals and five 150 mg/kg bw/day animals were found dead during the study period. All animals in the 150 mg/kg bw/day group exhibited salivation and wheezing. These signs were first noted during Week 2 and persisted throughout the duration of the treatment period. Other clinical signs noted primarily in this group included thinness, urine stains, alopecia, ocular opacity, rough haircoat, and red area on the face, ear, neck, leg(s), mouth, chest, and/or paw(s). Body weights for the 150 mg/kg bw/day males were generally lower than control values and corresponded with the lower food consumption values for this group. There was a 30% decrement in overall body weight gain when compared with control values. For the 150 mg/kg bw/day females, there was also a decrement in overall body weight gain (15%); however, it was less pronounced than in the males and there was no correlating lower total food consumption value.

Notable ophthalmoscopic findings observed in-life included posterior subcapsular or complete cataracts in two 30 mg/kg bw/day animals and twenty-one 150 mg/kg bw/day animals. The cataract noted for one of the animals in the 30 mg/kg bw/day group was attributed to a hyaloid remnant and is not considered to be related to the administration of the test material. Microscopically, the incidence of cataracts was noted to be 18/39 animals in the 150 mg/kg bw/day group. For the 30 mg/kg bw/day group, eyes were examined microscopically only from animals in which a gross lesion had been identified at necropsy, or from which cataracts had been diagnosed by the examining veterinary ophthalmologist. There were no microscopically identifiable cataracts in the mid-dose group.

Changes in clinical hematology parameters included significant increases in platelet count, leukocyte count, corrected leukocyte count, absolute segmented neutrophil count, and absolute lymphocyte count (females only) in the 150 mg/kg bw/day group. These increases were suggestive of a response to an inflammatory process. Changes in serum chemistry parameters were noted at the 150 mg/kg bw/day level and included a decrease in glucose (significant for the males) and albumin (significant for the males), and a significant increase in blood urea nitrogen for the female animals. These changes are considered to be attributable (either directly or secondarily) to the decline in nutritional status of this group. Additionally, there was a significant increase in gamma glutamyltransferase in the Group 4 animals.

Gross findings in the nonglandular stomach were noted in 4/40 animals in the 30 mg/kg bw/day group and 38/40 animals in the 150 mg/kg bw/day group. Findings included thickened mucosa, desquamation, and/or rough or pale mucosa. Microscopically, acanthosis was diagnosed in the nonglandular stomach in 6/40 animals in the 30 mg/kg/day group and 38/40 animals in the 150 mg/kg/day group.

Several mean absolute organ weights and/or organ-to-terminal- body-weight ratios were significantly different than control values for males and/or females of the 150 mg/kg bw/day group; however, there was no gross or microscopic correlate. Affected organs included the kidney, liver, adrenal, and testis/epididymis.

Inflammatory lung lesions were noted for 2/40 animals in the 30 mg/kg bw/day group and 10/40 animals in the 150 mg/kg bw/day group. These lesions are attributed to incidental aspiration of the test material.

In conclusion, based on the results of this study, Atmer 163, when administered orally via gavage to rats for 13 weeks at levels of 15, 30, and 150 mg/kg bw/day, yielded signs of toxicity at the mid- and high-dose levels. Based on these results, the no-observable-adverse-effect level (NOAEL) for Atmer 163 in Crl:CD®BR rats via gavage administration is 150 mg/kg bw/day for systemic effects and 15 mg/kg bw/d for local effects.

The corresponding dose finding study (Hazleton 564/161) that was conducted for 14 days with 30, 100 and 300 mg/kg bw/day, revealed an NOAEL of 100 mg/kg bw/day.

 

Another study was designed in a second species to characterize the potential subchronlc toxicity of a test material when administered to dogs for at least 13 weeks (Hazleton 564/164). The test material, Atmer 163, was administered via capsule to three groups of dogs (four dogs/sex/group) at dose levels of 15 (Group 2), 30 (Group 3), or 100 (Group 4) mg/kg bw/day. An additional group of four dogs/sex (Group 1) received an empty capsule daily and was thus considered the concurrent control group.

All dogs were observed twice daily for mortality and moribundity and once daily (approximately 2 to 3 hours postdose) for obvious indications of a toxic and/or pharmacologic effect. Individual body weights were recorded prior to initiation of dosing and weekly thereafter. Individual food consumption measurements and physical examinations were performed weekly. Indirect ophthalmoscopic examinations were performed on all dogs prior to initiation of dosing and on control and high-dose dogs prior to termination. Clinical pathology parameters (hematology and blood chemistry) were evaluated at termination. Following at least 13 weeks of administration of the test material, all dogs were humanely sacrificed and subjected to a complete gross necropsy. Organ weight evaluations and a histomorphologic examination of protocol-specified tissues were performed.

All dogs survived to the scheduled termination of the study. Clinical observations, apparently related to administration of the test material, included an increased incidence of salivation, emesis, and/or soft feces (mucoid only or mucoid/bilious) in the Group 4 males and/or females. Statistical evaluation of mean body weight, mean body weight gain, and mean food consumption values failed to demonstrate any significant differences when treated groups were compared to respective control values. Statistical evaluation of clinical pathology values revealed a significantly elevated mean erythrocyte value and a mean alanine aminotransferase value in the Group 4 females and a significantly elevated mean calcium value in the Group 3 and 4 females. In addition, the Group 3 male mean blood urea nitrogen value was significantly depressed as compared to the control value. Based on the small magnitude of change, absence of a dose response, or unusually low mean control value, the changes In calcium, blood urea nitrogen, and erythrocyte values, respectively, are felt to be incidental to treatment.

There were no apparent compound-related gross necropsy observations noted at termination. In addition, statistical evaluation of mean absolute and relative organ weight data failed to reveal any significant differences when treated groups were compared to the respective mean control values.

Histomorphologic examination of tissue sections revealed apparent compound-related changes in the livers of the Group 4 females, comprised of increased pigment accumulation noted in the Kupffer cells and bile canaliculi.

Based on the data generated from this study, the no-observable- adverse-effect level of Atmer 163 when administered via capsule for approximately 13 weeks to male and female beagle dogs is 30 mg/kg bw/day for systemic effects and 100 mg/kg bw/d for local effects.

The corresponding dose finding study (Hazleton 564/163) hat was conducted for 14 days with 10, 30 and 100 mg/kg bw/day, revealed an NOAEL of 100 mg/kg bw/day.


Justification for selection of repeated dose toxicity via oral route - systemic effects endpoint:
The selected study is the most adequate and reliable study with the lowest dose descriptor.

Justification for classification or non-classification

Based on the available data, Atmer 163 does not meet the classification criteria to classify for repeated dose toxicity according to Regulation (EC) 1272/2008 and therefore the data is conclusive, but not sufficient for classification.