N,N'-bis{N-[N-(4-chlorophenyl)carbamimidoyl]carbamimidoyl}hexane-1,6-bis(aminium) diundec-10-enoate

Acute Toxicity: oral

Administrative data

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

February 22, 2016 - May 11, 2016

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

test procedure in accordance with generally accepted scientific standards and described in sufficient detail

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2016

Report date:

2016

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test type:

acute toxic class method

Limit test:

no

Test material

Test material information

Constituent 1

Reference substance name:

N,N'-bis{N-[N-(4-chlorophenyl)carbamimidoyl]carbamimidoyl}hexane-1,6-bis(aminium) diundec-10-enoate

EC Number:

813-944-0

Cas Number:

1884575-91-0

Molecular formula:

C44H70Cl2N10O4

IUPAC Name:

N,N'-bis{N-[N-(4-chlorophenyl)carbamimidoyl]carbamimidoyl}hexane-1,6-bis(aminium) diundec-10-enoate

Test material form:

solid: particulate/powder

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Batch No.of test material: 001/CHUA/01 16
- Expiration date of the lot/batch: December 2017
- Purity test date: January 2016

Test animals

Species:

rat

Strain:

Wistar

Sex:

female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Animal breeding, RCC laboratories India Private Limited
- Females nulliparous and non-pregnant: [yes]
- Age at study initiation: 9-12 weeks
- Weight at study initiation: Females 158.3 to 206.1 g
- Fasting period before study: fasted overnight
- Housing:
- Diet (e.g. ad libitum):
- Water (e.g. ad libitum):
- Acclimation period: Under laboratory conditions for 7, 12, 15 and 8 days for Step I, Step II, Step III and Step IV, respectively after veterinary observation. Only animals without any visible signs of illness were used for the study.

ENVIRONMENTAL CONDITIONS
- Temperature (°C):
- Humidity (%):
- Air changes (per hr):
- Photoperiod (hrs dark / hrs light):

IN-LIFE DATES: From: To:

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

peanut oil

Details on oral exposure:

VEHICLE
- Concentration in vehicle: 300 mg and 2000 mg
- Amount of vehicle (if gavage): 10 mL
- Justification for choice of vehicle: Refined groundnut oil was selected as vehicle based on preliminary solubility testing which was
performed at the test facility before the study initiation date.


MAXIMUM DOSE VOLUME APPLIED: 10 mL/kg body weight

DOSAGE PREPARATION : The dose levels were in terms of the test item as supplied by the sponsor. The dose formulations were prepared shortly before each dosing. The test item (300 mg for Step I, II and 2000 mg for Step III, Step IV) was weighed into a pre-weighed glass beaker on a Sartorius balance. The test item was transferred to a mortar and triturated with pestle. Three mL of the vehicle was added to mortar and again triturated. 2 ml of vehicle was used twice to rinse the mortar and pestle. This was then transferred to a 10 mL volumetric flask. Sufficient quantity of vehicle was added to make up the required volume of suspension (w/v) to 10 mL. This was then transferred back to the beaker and mixed properly. The test item formed suspension in refined ground nut oil. Homogeneity of the test item in the vehicle was maintained during administration using a magnetic
stirrer.

CLASS METHOD (if applicable)
- Rationale for the selection of the starting dose:

Doses:

300 mg/kg body weight and 2000 mg/kg body weight

No. of animals per sex per dose:

Three animals / dose

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: daily
- Necropsy of survivors performed: yes
- Other examinations performed: clinical signs, body weight,organ weights, histopathology, other: mortality, viability, clinical signs

Statistics:

No statistics

Results and discussion

Effect levels

Key result

Sex:

female

Dose descriptor:

LD50

Effect level:

5 000 mg/kg bw

Based on:

test mat.

Mortality:

No mortality

Clinical signs:

other: No clinical signs

Gross pathology:

No abnormalities were observed in any of the treated animals of Step I (Animal No. 02, ;)~;; and Step II
(Animal No. 04, 05, 06) except Animal No. 01 revealed incidental finding on uterus which was
distended with watery content Moderately during necropsy at terminal sacrifice.
No abnormalities were observed in any of the 'Teated animals of Step III (Animal No 'Step N (Animal No. 10, 11, 12).

Other findings:

- Organ weights:
- Histopathology:
- Potential target organs:
- Other observations:

Any other information on results incl. tables

No mortalities were observed in any of the treated animals dosed at 300 and 2000 mg/kg body weight in Steps I, II, III and IV.

All the animals were normal throughout the acclimatization period. All the treated animals at 300 (Step I and Step II) and 2000 (Step III and Step IV) mg/kg body weight were found to be normal at approximately 30 minutes, at 1, 2, 3 and 4 hours observation an day 0 post

test item administration till the last day of observation period {day 14).

All the animals of Step I, Step II, Step III and Step IV had gained body weight by days 7 and 14 compared to day 0

Ne abnormalities were observed in any of the treated animals of Step I (Animal No. 02, 03) and Step II

(Animal No. 04, 05, 06) except Animal No. 01 revealed incidental finding on uterus which was distended with watery content Moderately during necropsy at terminal sacrifice. No abnormalities were observed in any of the 'treated animals of Step III (Animal No 07, 08, 09) and Step N (Animal No. 10, 11, 12).

Applicant's summary and conclusion

Interpretation of results:

Category 5 based on GHS criteria

Conclusions:

LD50 (cut off value): 5000 mg/kg body weight and is classified as category 5 or unclassified.
LD50 greater than 2000 mg/kg body weight.

Executive summary:

Four groups, each of three female Wistar rats, fasted overnight, were treated with CHUA by oral gavage administration at a dosage of 300 mg/kg body weight for Step I, II and 2000 mg/kg body weight for Step III, IV. The test item was formulated in vehicle (Refined groundnut oil) at a concentration of 30mg/mL (Step I, II) and 200 mg/mL (Step III, IV). The test item was administered at a dose volume of 10mL/kg body weight.

The animals were observed daily during the acclimatization period and mortality/viability and clinical signs were recorded. All the animals were observed for clinical signs during first 30 minutes and at approximately 1, 2, 3 and 4 hours post test item administration on test day 0 and once daily during test days 1-14. Mortality/viability was recorded during first 30 minutes and at approximately 1, 2, 3 and 4 hours post test item administration on test day 0 (in common with the clinical signs) and twice daily during days l-14 (once on day of sacrifice). Body weights were recorded on test days 0 (prior to test item administration), 7 and 14. All the animals were necropsied and examined macroscopically at the end of observation period.

The animals were treated as follows:

• Step I 3 females treated at 300 mg/kg, Mortality '0 out of 3'
• Step II 3 females treated at 300 mg/kg, Mortality '0 out of 3'
• Step III 3 females treated at 2000 mg/kg, Mortality '0 out of 3'
• Step IV 3 females treated at 2000 mg/kg, Mortality '0 out of 3'

All the animals appeared normal throughout the acclimatization period. No mortalities were observed in any of the treated animals dosed at 300 and 2000 mg/kg body weight in Steps I, II, III and IV.

All the treated animals at 300 (Step I and Step II) and 2000 (Step III and Step IV) mg/kg body weight were found to be normal at approximately 30 minutes, at 1, 2, 3 and 4 hours observation on day 0 post test item administration till the last day of observation period (day 14).

All the surviving animals of Step I, II, III and IV had gained body weight by days 7 and 14 compared to day 0.

No abnormalities were observed in any of the treated animals of Step I (Animal No. 02, 03) and Step II (Animal N^o. 04, 0'5, 06) except Animal N^o. 01 revealed incidental finding on uterus which was distended with watery content moderately during necropsy at terminal sacrifice.

No abnormalities were observed in any of the treated animals of Step III (Animal No. 07, 08, 09) and Step IV (Animal No. 10, 11, 12).