Carbamic acid, N-[[5-[[[2-[2-(dimethylamino)ethoxy]ethoxy]carbonyl]amino]-1,3,3-trimethylcyclohexyl]methyl]-, 2-[2-(dimethylamino)ethoxy]ethyl ester

Repeated dose toxicity: oral

Administrative data

Endpoint:

short-term repeated dose toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

13 Mar - 29 Apr 2002

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP Guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2002

Report date:

2002

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

EU Method B.7 (Repeated Dose (28 Days) Toxicity (Oral))

Version / remarks:

EEC Directive 96/54, B.7

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

OECD Guideline 407 (Repeated Dose 28-Day Oral Toxicity Study in Rodents)

Deviations:

no

Test guideline 3

Qualifier:

according to guideline

Guideline:

other: Japanese Guidelines for Screening, Toxicity Testing of Chemicals: Testing Methods for new Substances, enacted July 13, 1974, amended Dec 5, 1986

GLP compliance:

yes

Limit test:

no

Test material

Test material information

Constituent 1

Reference substance name:

-

EC Number:

441-100-8

EC Name:

-

Cas Number:

351197-46-1

Molecular formula:

Hill formula: C24 H48 N4 O6 CAS formula: C24 H48 O6 N4

IUPAC Name:

2-[2-(dimethylamino)ethoxy]ethyl N-{[1,3,3-trimethyl-5-(9-methyl-2-oxo-3,6-dioxa-1,9-diazadecan-1-yl)cyclohexyl]methyl}carbamate

Constituent 2

Reference substance name:

Carbamic acid, [[5-[[[2-[2- (dimethylamino)ethoxy]ethoxy]carbonyl]amino]-1,3,3- trimethylcyclohexyl]methyl]-,2-[2- (dimethylamino)ethoxy]ethyl ester

IUPAC Name:

Carbamic acid, [[5-[[[2-[2- (dimethylamino)ethoxy]ethoxy]carbonyl]amino]-1,3,3- trimethylcyclohexyl]methyl]-,2-[2- (dimethylamino)ethoxy]ethyl ester

Details on test material:

- Name of test material (as cited in study report): UAX-1179
- Physical state / appearance: Liquid / pale amber
- Analytical purity: >99% proprietary tertiary amines
- Lot/batch No.: M. 17-08-01
- Expiration date of the lot/batch: 19 Sept 2003
- Storage condition of test material: In the original container under nitrogen at room temperature (17 - 23 °C), keep container closed and away from heat and flame
- Safety precautions: Routine hygienic procedures (gloves, goggles, face mask)

Test animals

Species:

rat

Strain:

other: HanBrl: Wistar (SPF)

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: RCC Ltd Biotechnology & Animal Breeding Division, CH-4414 Fijllinsdorf / Switzerland
- Age at delivery: 6 weeks
- Body weight range at acclimatization: Males: 130 - 158 g, Females: 109 - 130 g
- Housing: In groups of five in Makrolon type-4 cages with wire mesh tops and standardized softwood bedding ('Lignocel' Schill AG, CH-4132 Muttenz/Switzerland).
- Diet (e.g. ad libitum): Pelleted standard Provimi Kliba 3433 (batch nos. 1 19/01 and 05/02) rat maintenance diet (Provimi Kliba AG, CH-4303 Kaiseraugstl Switzerland) was available ad libitum.
- Water (e.g. ad libitum): Community tap-water from ltingen was available ad libitum in water bottles.
- Acclimation period: one week

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 +/- 3 °C
- Humidity (%): 30 - 70%
- Air changes (per hr): 10 - 15
- Photoperiod (hrs dark / hrs light): 12 (12 hours fluorescent light / 12 hours dark)
- music during the light period

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

other: bidistilled water

Details on oral exposure:

Method of administration: gavage
Dose volume: 10 mL/kg bw

Analytical verification of doses or concentrations:

not specified

Details on analytical verification of doses or concentrations:

Concentration, homogeneity and stability (after 2 hours and 7 days) of the dose formulations were determined in samples taken after experimental start. Concentration and homogeneity of the dose formulations were determined in samples taken during week 3 of the treatment. The analyses were performed by RCC Ltd (Environmental Chemistry & Pharmanalytics Division) according to a HPLC method supplied by the Sponsor.

Duration of treatment / exposure:

Test duration: 28 days

Frequency of treatment:

Dosing regime: 7 days/week

Doses / concentrations

Remarks:

Doses / Concentrations:
0, 30, 150, 450 mg/kg bw/day
Basis:
actual ingested

No. of animals per sex per dose:

at dose 0 mg/kg bw/day : 10 males and 10 females
at dose 30 mg/kg bw/day : 5 males and 5 females
at dose 150 mg/kg bw/day : 5 males and 5 females
at dose 450 mg/kg bw/day : 10 males and 10 females

The recovery groups include the half of group 1 (0 mg/kg bw) and 4 (450 mg/kg bw)

Control animals:

yes, concurrent vehicle

Details on study design:

- Dose selection rationale: Based upon the results of a non-GLP 5-day dose-range- finding study (RCC Study Number 838440) in which UAX-1179 was administered by gavage to 2 rats per group and sex.

Positive control:

no

Examinations

Observations and examinations performed and frequency:

Observations for mortality/viability were recorded twice daily.

CAGE SIDE OBSERVATIONS: Yes
- Time schedule: The animals were observed for clinical signs once before commencement of administration; twice daily on days 1-3; as well as once daily on days 4-28, and once daily during days 29-42 (recovery).
Appearance: Piloerection, Salivation, Hunched posture
Motor: Ataxia, Tremor/twitching, Prostration, Circling, Spasm
Behavior: Hyperactivity, Somnolence, Increased exploration, Reduced grooming, Vocalisation
Respiration: Dyspnea, Tachypnea, Bradypnea
Reflexes: Blink, Pinna, Iridic light reflex, Push-off (hind leg), Pain response, Startle/hearing
Miscellaneous: Lacrimation, Limbs cyanotic, Mydriasis, Miosis, Exophthalmos, Reduced muscle tone

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: weekly; The animals were observed in their home cages, outside their home cages in a standard arena and in the hand. These observations were performed in random sequence once before commencement of administration and once weekly (weeks 1-3) thereafter.

BODY WEIGHT: Yes
- Time schedule for examinations: Body weights were recorded weekly during pretest, treatment and recovery and before necropsy.

FOOD CONSUMPTION:
- The food consumption was recorded once during the pretest period and weekly thereafter.

HAEMATOLOGY: Yes
Paramters: Erythrocyte count, Hemoglobin, Hematocrit, Mean corpuscular volume, Red cell volume distribution width, Mean corpuscular hemoglobin, Mean corpuscular hemoglobin concentration, Hemoglobin concentration distribution width, Reticulocyte count, Reticulocyte maturity index, Leukocyte count, Differential leukocyte count, Neutrophils, Eosinophil, Basophil, Lymphocytes, Monocytes, Large unstained cells, Thrombocyte count, Methemoglobin, Heinz bodies, Prothrombin time, Partial thromboplastin time

CLINICAL CHEMISTRY: Yes
Glucose, Urea, Creatinine, Bilirubin total, Cholesterol, Triglycerides, Phospholipids, Aspartate aminotransferase, Alanine aminotransferase, Lactate dehydrogenase, Creatine kinase, Alkaline phosphatase, Gamma-glutamyl transpeptidase, Sodium, Potassium, Chloride, Calcium, Phosphorus inorganic, Protein total, Albumin, Globulin, Albumin/ Globulin Ratio

URINALYSIS: Yes
Urine volume (18 h), Relative Density, Osmolality, Color, Appearance,
pH-value, Nitrite, Protein, Glucose, Ketones, Urobilinogen, Bilirubin, Erythrocytes, Leukocytes, Cells, Casts, Crystals

NEUROBEHAVIOURAL EXAMINATION: Yes
- Battery of functions tested: during week 4, relevant parameters from a modified Irwin screen test were evaluated in all animals.

OTHER: Grip strength and locomotor activity were tested.

Sacrifice and pathology:

GROSS PATHOLOGY: Yes
All animals were weighed and necropsied. Descriptions of all macroscopic abnormalities were recorded. Samples of the following tissues and organs were collected from all animals at necropsy and fixed in neutral phosphate buffered 4 % formaldehyde solution:
Adrenal glands, Aorta, Bone (sternum, femur including joint), Bone marrow (femur), Brain (cerebrum, cerebellum, brain stem), Cecum, Colon, Duodenum, Epididymides (fixed in Bouin's solution), Esophagus, Eyes with optic nerve (fixed in Davidson's solution), Harderian gland (fixed in Davidson's solution), Heart, Ileum, with Peyer's patches, Jejunum with Peyer's patches, Kidneys, Larynx, Lacrimal gland (exorbital), Liver, Lungs (infused with formalin at necropsy), Lymph nodes (mesenteric, mandibular), Mammary gland area, Nasal cavity, Ovaries, Pancreas, Pituitary gland, Prostate gland, Rectum, Salivary glands (mandibular, sublingual), Sciatic nerve, Seminal vesicles, Skeletal muscle, Skin, Spinal cord (cervical, midthoracic, lumbar), Spleen, Stomach, Testes (fixed in Bouin's solution), Thymus, Thyroid (incl. parathyroid gland), Tongue, Trachea, Urinary bladder (infused with formalin at necropsy), Uterus, Vagina, Gross lesions.
The following organ weights were recorded on the scheduled dates of necropsy:
Brain, Heart, Liver, Thymus, Kidneys, Adrenals, Spleen Ovaries, Testes, Epididymides

HISTOPATHOLOGY: Yes
Slides of all organs and tissues listed which were collected at scheduled sacrifice from the animals of control and high-dose groups were
examined by a pathologist.
All organ and tissue samples, as defined under Histopathology, were processed, embedded and cut at an approximate thickness of 2 to 4 micrometers, and stained with hematoxylin and eosin.

Statistics:

The following statistical methods were used to analyze the grip strength, locomotor activity, body weight, organ weights and ratios, as well as:
The Dunnett-test (many to one t-test) based on a pooled variance estimate will be applied if the variables can be assumed to follow a normal distribution for the comparison of the treated groups and the control groups for each sex.
The Steel-test (many-one rank test) will be applied instead of the Dunnett-test when the data can not be assumed to follow a normal distribution.
Student's t-test was applied to grip strength and locomotor activity.
Fisher's exact-test will be applied to the macroscopic findings.
For clinical laboratory data, quantitative data were analyzed by a one-way analysis of variance (ANOVA) when the variances are considered homogeneous according to Bartlett. Alternatively, if the variances are considered to be heterogenous (p<= 0.05), a non-parametric Kruskal-Wallis test will be used. Treated groups were compared to the control groups using Dunnett's test if the ANOVA was significant at the 5% level and by Dunn's test in the case of a significant Kruskal-Wallis test (p<= 0.05).

References :
C.W. Dunnett: A Multiple Comparison Procedure for Comparing Several Treatments with a Control, J.
Amer. Stat. Assoc. 50, 1096-1 121 (1955).
S.C. Gad and C.S. Weil: Statistics and Experimental Design for Toxicologists. The Telford
Press, Caldwell, New Jersey, 43-45 (1986).
W.H. Kruskal and W.A. Wallis: Use of ranks in one-criterion variance analysis. Journal of the American
Statistical Association, 47, 583-621 (1952).
O.J. Dunn: Multiple comparisons using rank sums. Technometrics 6, 241 -252 (1964).
R.G. Miller: Simultaneous Statistical Inference, Springer Verlag, New York (1981).
R.A. Fisher: Statistical Methods for Research Workers, Oliver and Boyd, Edinburgh (1950).

Results and discussion

Results of examinations

Details on results:

Clinical observations:
MORTALITY / VIABILITY

One male treated with 450 mg/kg/day died at treatment day 23 and one male of the same dose group at day 26. Four females treated with 450 mg/kg/day died on treatment days 5, 10 and 19. The cause of the deaths was not evident at microscopic level. However, a relation to the treatment with the test item cannot be excluded. All other animals survived until scheduled necropsy.


CLINICAL SIGNS

No test item related clinical signs were evident in animals treated with 30 or 150 mg/kg/ bw/day. The following signs were observed in animals treated with 450 mg/kg/day: Slight piloerection was seen in two males and three females in treatment week three and in one female in treatment week four. These findings may be test item related. Slight ataxia was observed in two males and three females in treatment week three and in one female in treatment week two. Slight tremor was observed in two males and five females in treatment week three and in one female in treatment week two. It could not be excluded that these findings were test item related. Slight spasm was noted in two females in treatment week one. It could not be excluded that this was test item related. Slight sedation was noted in two males and seven females in treatment week three and one female in treatment week two. It could not be excluded that this was test item related. Slight dyspnea was noted in four females in treatment week one. Slight bradypnea was noted in one male in treatment week one. Breathing noise was evident in two males and in four females in treatment week three. It could not be excluded that these findings are due to test item treatment. No other clinical signs were evident in males or females (week 1-3).

The following test item related findings were noted in males or females treated with 450 mg/kg/day:
Moderate to slight alopecia was noted in one male from the pretest until treatment day nine. Slightly pale feces was observed in all surviving males and females at 450 mg/kg/day from treatment day 7 until the first day of the recovery period. This was considered to be a passive effect of the test item with no toxicological relevance. Slight piloerection was seen on treatment day 26 in four of eight males and in two of six females and in one of eight males as well as in one of six females on day 7. This finding was considered to be test item related. Slight tremor was noted in two males on day 26 and in one male on days 27 and 28. This finding was noted in one female on days 4 and 5, three females on day 26, four females on day 27 and one female on day 28. Slight sedation was observed in five, three or one of eight males and in two, one or one of six females on days 26, 27, or 28, respectively. Abnormal breathing (dyspnea or bradypnea) as well as breathing noises were noted in several males and some females. Although transient this finding was considered to be test item related.
Hunched posture was observed in one male on day 28. Slight prostration was observed in one male and one female on day 26. Although transcient these findings were considered to be test item related. No other clinical signs were evident in animals treated with 450 mg/kg/day.


FUNCTIONAL OBSERVATIONAL BATTERY

Grip Strength

No test item related changes in fore- or hind limb grip strength were evident in males or females when compared with controls.

Locomotor Activity

Significantly decreased (p<0.01) locomotor activity was observed in males treated with 450 mg/kg/day during the whole observation period of 60 minutes when compared with controls. In females of the same dose group this activity was significantly decreased in the first 30 minutes (p<0.01), and between 45 and 60 minutes (p<0.05) of observation. Between 30 and 45 minutes this decrease was not significant. Significantly decreased locomotor activity was also observed in males treated with 150 mg/kg/day in the first 30 minutes of observation (p<0.01), and between 45 and 60 minutes (p<0.05). Between 30 and 45 minutes this decrease was not significant. These differences in locomotor activity were considered to be test item related.

No further changes in locomotor activity were noted when compared with controls.


FOOD CONSUMPTION

No changes in mean daily or relative food consumption were observed in males or females after four or six weeks when compared with control animals.


BODY WEIGHT

Significantly decreased (p<0.05) mean body weight was noted in males treated with 450 mg/kg/day in treatment week four when compared with the controls. Significantly decreased (p<0.05) mean body weight gain was observed in males treated with 450 mg/kg/day in treatment weeks three and four when compared with the controls. Significantly (p<0.01) decreased mean body weight gain was noted in females treated with 450 mg/kg/day on days 22 (week four) and 28 (week four) and in females treated with 150 mg/kg/day on day 28 (week four). Significantly decreased (p<0.05) mean body weight gain was noted in females treated with 450 mg/kg/day within week two of the recovery period when compared with controls. These effects were considered to be test item related. No other significant changes were observed in males or females after four or six weeks.

Laboratory findings:
Hematology

Significantly decreased relative and absolute reticulocytes were noted in males treated with 450 mg/kg/day after four weeks when compared with controls. A dose response relationship could be observed within the absolute values. The decrease was also observed in females of this dose group but without statistical significance after four or six weeks when compared with controls. It can not be excluded that this effect is test item related. No other test item related changes in parameters of hematology were seen in test item treated animals after four or six weeks when compared with controls.


Clinical Biochemistry

Significantly increased activity of Aspartate aminotransferase was measured in males treated with 150 and 450 mg/kg/day after four and six weeks when compared with controls. This increase was also observed in females but was only significant at 450 mg/kg/day. Due to this and due to dose response relationship the increase was considered to be test item related.

The activity of Lactate dehydrogenase was significantly increased in males treated with 150 or 450 mg/kg/day after four weeks when compared with controls. This was considered to be test item related because it was also seen in males treated with 450 mg/kg/day after six weeks and in females treated with 450 mg/kg/day after four weeks when compared with controls.


Urinalysis

No test item related changes in parameters of urinalysis were evident in males or females after four or six weeks when compared with control animals

The volume was significantly decreased (p<0.05) in males treated with 450 mg/kg/day after four weeks when compared with the controls. This finding was considered to be incidental because it was seen in males only.
A significantly increased mean value for ketones was noted in females treated with 450 mg/kg/day after four weeks when compared with the controls. This finding was considered to be incidental because the value lies within the range of the historical control data.
The pH value was significantly decreased in males treated with 450 mg/kg/day after six weeks when compared with controls. This finding was considered to be incidental because this value lies within the range of the historical control data.
No other changes in urinalysis parameters were noted in males or females after four or six weeks when compared with the controls.

Effects in organs:
ORGAN WEIGHTS

No changes in mean organ weights, organ to body- or organ to brain weight ratios were noted in test item treated males after four weeks when compared with controls.

Significantly increased (p<0.01) mean liver to body weight ratios were noted in females treated with 150 or 450 mg/kg/day after four weeks when compared with controls. This was considered to be test item related because a dose response relationship was observed.

No test item related changes in mean organ weights, organ to body- or organ to brain weight ratios were noted in males and females treated with 450 mg/kg/day after six weeks when compared with controls.

Significantly decreased (pe0.05) mean thymus weight and heart to brain weight ratios were noted in females treated with 450 mg/kg/day when compared with controls. These effects were considered to be incidental because no dose-response relationship was observed and the finding was seen in one sex only.
Significantly increased (p<0.05) mean brain to body weight ratios were noted in females treated with 450 mg/kg/day. This finding was considered to be incidental because it was likely related to the decreased body weights of the females at this dose level.
Significantly decreased (p<0.05) mean thymus weights, mean thymus to brain weight ratios, mean liver to body weight ratios, and mean heart to brain weight ratios were noted in males treated with 450 mg/kg/day.
These findings were considered to be incidental because they were only seen in males and did not occur after four weeks.

MACROSCOPIC / MICROSCOPIC FINDINGS

A limited number of findings noted in the 450 mg/kg/day dose group only distinguished animals that died spontaneously from animals that survived to the scheduled necropsy:
Reddish to dark red discoloration of the: lung (1 male, 1 female), pancreas (1 male), mandibular lymph node and salivary glands (1 male)
Dark red focus/foci of the: lung (2 females), ovaries (1 female), thymus (2 males, 1 female)
intestinal tract distended with gas (1 female)
Dark red foci were also noted on the fundus of the stomach in a female animal that survived to the scheduled necropsy.

A number of findings were noted at the end of the treatment period. From these findings, the following ones distinguished decedent rats from rats at terminal sacrifice:
Lung, adrenal glands, stomach, liver, kidneys, thymus, mesenteric and mandibular lymph nodes, ovaries, brain, pancreas, salivary glands: congestion was present in some males and/or females treated at 450 mg/kg
Lung, thymus:hemorrhage occurred each in one female treated at 450 mg/kg.
Lung: alveolar edema was noted in one female treated at 450 mg/kg.
Microscopically, these congestions and/or hemorrhages in the various organs and a pulmonary edema were noted at the termination of the treatment period in decedents and were considered to be associated with the spontaneous deaths occurring in these animals.
A number of findings were diagnosed in the other organs and tissues examined at the termination of the treatment period. Their incidence, severity and morphologic appearance did not distinguish treated rats from the control rats.
A number of findings were noted at the end of the recovery period. From these findings, the following ones distinguished decedent rats from rats at terminal sacrifice: Salivary glands: congestion was present in one male treated at 450 mg/kg.

Effect levels

1

Dose descriptor:

NOAEL

Effect level:

150 mg/kg bw/day (nominal)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: decreased locomotor activity in males, decreased body weight gain in females

2

Dose descriptor:

NOEL

Effect level:

30 mg/kg bw/day (nominal)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: no test item related effects

3

Dose descriptor:

LOAEL

Effect level:

450 mg/kg bw/day (nominal)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: transient, slight clinical signs, decreased locomotor activity, decreased mean body weight gain and a number of gross findings (in males and females)

Target system / organ toxicity

Critical effects observed:

not specified

Any other information on results incl. tables

Table 1: Locomotor activity, summary, low beams count, males and females, week 4

Week 4		15 min.	30 min.	45 min.	60 min.	Total
Group1	Males	1005	544	296	407	2251
	Females	873	504	223	251	1850
Group 2	Males	844	459	406	378	2087
	Females	1114	390	366	341	2211
Group 3	Males	272**	242**	159	264*	936**
	Females	1021	438	159	239	1856
Group 4	Males	315**	164**	113**	137**	729**
	Females	209**	159**	133	38*	538**

*/** T-Test sig. at 5% or 1% level

Table 2: Body weight gain (%), summary, males and females, mean values

Mean body weight gain (%)		Group 1 0 mg/kg bw/d	Group 2 30 mg/kg bw/d	Group 3 150 mg/kg bw/d	Group 4 450 mg/kg bw/d
Day 1 week 1	Males	0	0	0	0
	Females	0	0	0	0
Day 8 week 2	Males	21	21	24	18
	Females	16	15	16	14
Day 15 week 3	Males	37	36	39	29*
	Females	27	25	25	22
Day 22 week 4	Males	47	49	50	38
	Females	38	36	34	33**
Day 28 week 4	Males	59	56	59	42*
	Females	44	42	35**	35**

*/**: Dunnett-Test based on pooled variance significant at 5% (*) or 1% (**) level

Table 3a: Macroscopical findings, summary, males

MALES	Group 1		Group 2		Group 3		Group 4		Group 1		Group 4
After 4 weeks									After 6 weeks
Animals examined	5		5		5		5		5		5
Animals without findings	5		5		5		3		5		4
Stomach: Focus/Foci	0	0%	0	0%	0	0%	1	20%
Pancreas: Discoloration	0	0%	0	0%	0	0%	1	20%
Thymus: Focus/Foci	0	0%	0	0%	0	0%	1	20%	0	0%	1	20%
General observations: Beginning autolysis									0	0%	1	20%
Lungs: Discoloration									0	0%	1	20%
Ileum: organ missing									0	0%	1	20%
Thymus: Focus/Foci									0	0%	1	20%
Mandibular L. Node: Discoloration									0	0%	1	20%
Salivary glands: Discoloration									0	0%	1	20%

empty = without findings (0 / 0%)

Table 3b: Macroscopical findings, summary, females

FEMALES	Group 1		Group 2		Group 3		Group 4		Group 1		Group 4
After 4 weeks									After 6 weeks
Animals examined	5		5		5		5		5		5
Animals without findings	5		3		4		2		3		5
Lungs: Discoloration Foamy fluid released from bronchi Focus/Foci	0	0%	0	0%	0	0%	1	20%	1	20%	0	0%
	0	0%	0	0%	0	0%	1	20%
	0	0%	0	0%	0	0%	2	40%
Jejunum distended with gas	0	0%	0	0%	0	0%	1	20%
Ileum distended with gas Organ missing	0	0%	0	0%	0	0%	1	20%
	0	0%	0	0%	0	0%	1	20%
Caecum: distended with gas	0	0%	0	0%	0	0%	1	20%
Kidneys: Pelvic dilation	0	0%	2	40%	0	0%	0	0%	1	20%	0	0%
Ovaries: Focus/Foci Reduced in size	0	0%	0	0%	0	0%	1	20%
	0	0%	0	0%	1	20%	0	0%
Thymus: Focus/Foci	0	0%	0	0%	0	0%	1	20%

empty = without findings (0 / 0%) Table 4: Weekly outside cage observations, summary data (only affected points), males / females

MALES / FEMALES

Group 1

Group 2

Group 3

Group 4

Number of Animals in group

10

10

10

10

10

5

5

10

10

10

10

9

Findings/ study week

score

P

1

2

3

4

P,1-4

P,1-4

P

1

2

3

4

APP

Piloerection

1-3

0/2-

2-/4-

5-/3-

Alopecia, neck, r.

1-3

1-/0

1-/0

MOT

Ataxia

1-3

0/1-

2-/3-

Tremor/twitching

1-3

0/1-

2-/5-

3-/3-

Prostration

1

2-/2-

Spasm

1-3

0/2-

BEH

Sedated

1-3

0/1-

2-/7-

6-/2-

RES

Dyspnea

1

0/4-

2-/2-

Bradypnea

1

1-/0

Breathing noise

1-3

2-/4-

0/2-

MIS

Raddish scab, r. eye

1-3

1-/0

1-/0

1-/0

1-/0

1-/0

Abbreviations (1a and 1b): APP = Appearance, MOT = Motor, BEH = Behavioral, RES = Respiration, MIS = Miscellaneous, P = Pretest examination (during acclimatization) number in squares= Number of affected animals, - = mean severity between 1.00 and 1.67, + = mean severity between 1.68 and 2.33, ! = mean severity between 2.34 and 3.00

Applicant's summary and conclusion

Conclusions:

The NOEL (oral, rat, 28 days) is considered to be 30 mg/kg bw/day .
The NOAEL (oral, rat, 28 days) is considered to be 150 mg/kg bw/day.

Executive summary:

Oral administration of UAX-1179 to Wistar rats at doses of 30, 150 and 450 mg/kg bw/day, for 28 days resulted in mortality of two males and four females treated with 450 mg/kg bw/day and no effects upon food consumption, grip strength and urinanalysis. A total of 10 females and 10 males were treated with 450 mg/kg bw/day or left untreated. Respectively 5 animals per sex/dose were treated at 30 and 150 mg/kg bw daily/28 days.

Treatment-related findings were generally restricted to various transient, slight clinical signs (slight piloerection, slight ataxia, slight tremor, slight spasm, slight sedation, slight dyspnea, slight bradypnea and breathing noise daily and/or weekly) at 450 mg/kg bw/day, significantly decreased locomotor activity at 150 or 450 mg/kg bw/day at different time points, significantly decreased mean body weights and body weight gain in males treated with 450 mg/kg bw/day and significantly decreased body weight gain in females at 150 and 450 mg/kg bw/day, and a number of gross findings distinguishing some treated decendents from rats at terminal sacrifice.

Based on these results the NOEL of UAX-1179 is considered to be 30 mg/kg body weight/day and 150 mg/kg body weight/day the NOAEL.