Registration Dossier

Administrative data

Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Guideline study and GLP

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2004
Report Date:
2004

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method)
Deviations:
no
GLP compliance:
yes (incl. certificate)
Test type:
acute toxic class method
Limit test:
yes

Test material

Reference
Name:
Unnamed
Type:
Constituent
Type:
Constituent
Details on test material:
batch No.: Kn-Gi 8634/1
KN 172
SAT 030678

Test animals

Species:
rat
Strain:
Sprague-Dawley
Sex:
male/female
Details on test animals and environmental conditions:
Healthy rats of the Hsd: Sprague Dawley SD strain were ordered and obtained from Harlan
Italy S.r.l., 33049 San Pietro al Natisone (UD), Italy. Animals were ordered weighing 176 to
200 grams and aged approximately 6 to 8 weeks with female animals nulliparous and nonpregnant.
Animals appeared to be in an acceptable condition following arrival in different
batches on 9. and 23.April 2004. A pre-dose acclimatisation period of at least 5 days was allowed.
Animals included in the study were housed, in groups of 3 animals of the Same Sex, in
polycarbonate cages measuring 42.5x26.6x18 cm and equipped with a stainless steel mesh lid
and floor. Cages were suspended over trays holding an absorbent material which was
inspected daily and changed as necessary. Throughout the study each cage was identified by a
colour coded labe1 recording the study number, animal number and the details of treatment.
Animal room controls were set to maintain temperature within the range of 22°C +- 2°C and
relative humidity within the range of 55% +-15%. Actual conditions were recorded.
The room was lit by fluorescent tubes controlled to give an artificial cycle of 12 hours light
and 12 hours dark each day.

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
water
Details on oral exposure:
On the day of dosing, the amount of the formulated test item to be administered was calculated
for each fasted animal according to body weight. This was administered, by gavage atallowed. a dose
volume of 10 mlkg, using a rubber catheter attached to a syringe of suitable capacity.
Food was made available approximately 4 hours after dosing.
Doses:
50, 300, 2000 mg/kg bw
No. of animals per sex per dose:
3
Control animals:
no
Details on study design:
Single groups of 3 females were dosed at a level of 2000 and 300 mgkg (steps 1 and 2).
One group of 3 females and 1 group of 3 males were subsequently dosed at a level of 50
mgkg.
Selection and allocation
The required number of animals for the study was allocated to treatment groups. Individuals
were permanently identified following arrival by a combination of ear notch (units) and tattoo
on the feet. Male animals were identified by even numbers and females by odd numbers. The
body weight of each individual was withn 20% of the mean.
Groups of 3 animals were allocated to the study as fo1lows:

Dose level Step I Anima1 number
(mgkg) (Males Females)

2000 337,339,341
300 355,357,359
50 379,381,383300


On the day of dosing, the amount of the formulated test item to be administered was calculated
for each fasted animal according to body weight. This was administered, by gavage at a dose
volume of 10 mlkg, using a rubber catheter attached to a syringe of suitable capacity.
Food was made available approximately 4 hours after dosing.
Mortality and morbidity
Throughout the study all animals were checked twice daily.
Clinical signs
Animals were observed for clinical signs immediately upon dosing, approximately 30 minutes,
2 and 4 hours after dosing and daily thereafter for a total of 14 days, where appropriate.
Body weight
All animals were weighed at allocation to the study (Day -I), immediately prior to dosing
(Day 1) and on Days 2, 8 and 15, where appropriate. Early decedent animals were weighed
when found dead.

Termination
Surviving animals were killed on Day 15 by carbon dioxide narcosis.
All animals, including those found dead, were subjected to a goss necropsy examination for
both external and internal abnormalities. The cranial, thoracic and abdominal cavities were
opened to allow examination of their contents. Larger Organs were sectioned. Both the
stomach and representative sections of the gastro-intestinal tract were opened for examination of the mucosal surfaces.

Results and discussion

Effect levels
Sex:
female
Dose descriptor:
LD50
Effect level:
50 - 300 mg/kg bw
Mortality:
Following dosing a single group of 3 female animals at 2000 mgkg (step l), one animal was
found dead within 30 minutes of dosing and another was found dead on Day 2. Lethargy was
noted in all animals on the day of dosing. Hunched posture was observed in the surviving
animal on Day 2 and piloerection from Day 2 to Day 4. Complete recovery had occurred by
Day 5. No mortality occurred following dosing in the two groups of 3 female and 3 male animals at
50 mgkg .
Clinical signs:
Following dosing the 3 female animals at 300 mgkg (step 2), one animal died within 4 hours
of dosing and the remaining two animals were found dead on Day 2. Rales and hunched
posture were noted on the day of dosing. Animals were observed for clinical signs immediately upon dosing, approximately 30 minutes,
2 and 4 hours after dosing and daily thereafter for a total of 14 days, where appropriate. No clinical signs were noted, with the exception of a damaged left ear which was observed in 1 male animal on Day 15.
Body weight:
Changes in body weight observed during the period of the study in the surviving animals were
within the range expected for this strain and age of animal.
Gross pathology:
Abnormal contents were found in the stomach (clear or yellow fluid material) of 2 females
dosed at 2000 mgkg and 2 females dosed at 300 mgkg. Abnormal contents (red, mucoid
material) were noted in the jejunum and ileum of a single animal dosed at 2000 mgkg.
Extemal examination revealed brown staining of the skidfw around the wo-genital region in
a single early decedent animal.
No abnormalities were observed in any animal killed on termination of the study.
Other findings:
Termination
Surviving animals were killed on Day 15 by carbon dioxide narcosis.
All animals, including those found dead, were subjected to a goss necropsy examination for
both external and internal abnormalities. The cranial, thoracic and abdominal cavities were
opened to allow examination of their contents. Larger Organs were sectioned. Both the
stomach and representative sections of the gastro-intestinal tract were opened for examination of the mucosal surfaces.

Applicant's summary and conclusion

Interpretation of results:
toxic
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
The acute toxicity of KN 172 was investigated following administration of a single oral dose
to the rat.
Mortality occurred following dosing at 2000 and 300 mg/kg. No mortality or other signs of
toxicity were observed in animals dosed at 50 mg/kg.
These results indicate that the test item, KN 172, has a toxic effect in the rat following single
oral administration at dose levels of 2000 and 300 mg/kg. The mortality pattern demonstrates
the LD50 to be less than 300 mg/kg, but greater than 50 mg/kg body weight.


Executive summary:

The acute toxicity of KN 172 was investigated following administration of a single oral dose

to the rat.

A single group of 3 female animals was dosed at a level of 2000 mgkg and obsemed for a

period of 14 days (step 1). Two animals died on days 1 and 2. Clinical signs included lethargy,

hunched posture and piloerection. Complete recovery had occurred in the surviving animal by

Day 5.

An additional 3 female animals were then dosed at 300 mgkg (step 2). All animals died

following dosing. Clinical signs noted on the day of dosing were hunched posture and rales.

Three female animals were subsequently dosed at 50 mg/kg and observed for a period of 14

days (step 3). No mortality occurred and no clinical signs were observed following dosing.

Three male animals were finally dosed at 50 mgkg and observed for a period of 14 days (step

4). No mortality occurred and no clinical signs were obsemed following dosing.

Surviving animals were killed at the end of the observation period. All animals were subjected

to necropsy examination.

Changes in body weight obsemed were not remarkable.

Abnormal contents in the jejunum, ileum and/or in the stomach were observed in the early

decedent animals. Skinlfur staining was also noted in a single early decedent animal. No

abnormalities were obsemed in any animal killed on termination of the study.

These results indicate that the test item, KN 172, has a toxic effect in the rat following oral

administration of single dose levels of 2000 and 300 mgkg. No mortality or other signs of

toxicity were observed at 50 mgkg/day. The mortality pattem demonstrates the LD50 to be

less than 300 mgkg, but greater than 50 mgkg body weight.

European Directives conceming the classification, packaging and labelling of dangerous

substances (6715481EEC and subsequent revisions) would suggest the fol1owing:-

Classification : Required

Symbol : T

R phrase : R25 -Toxic if swallowed