N6-(1-oxododecyl)-L-lysine

Endpoint summary

Description of key information

The aim of this repeated dose toxicity study was to obtain information on the toxicity of Famex LL T administered once daily by oral administration via gavage to rats for 28 consecutive days. The animals were treated with 100, 300 or 1000 mg Famex LL T/kg b.w./day. The control animals received the vehicle (0.5% Methylcellulose 400). None of the animals died or had to be sacrificed prematurely. No test itemrelated changes were observed for the behaviour or external appearance of the animals, the detailed clinical observations, the neurological screening, the body weight, body weight gain and body weight at autopsy, the food and drinking water consumption, for any of the haematological and clinical chemical parameters, the eyes or optic region, the auditory acuity, the relative and absolute organ weights, and at macroscopic inspection at necropsy at any dose level. The histopathological examination did not reveal any test item-related morphological changes. The experimental no-observed-effect level (NOEL) was above 1000 mg Famex LL T/kg b.w./day, by daily oral administration.

Key value for chemical safety assessment

Repeated dose toxicity: via oral route - systemic effects

Link to relevant study records

Reference

Endpoint:

short-term repeated dose toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2017

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

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

Version / remarks:

2008

Deviations:

no

Qualifier:

according to guideline

Guideline:

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

Version / remarks:

2008

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Limit test:

no

Specific details on test material used for the study:

Designation: Famex LL T
Batch no.: 130918
Characteristics: White to pale yellow powder
Storage conditions: At +10°C to +25°C
Content: Main component: 99.9%; Accessory component: Water 0.1%

Species:

rat

Strain:

CD-1

Remarks:

Crl:CD(SD)

Details on species / strain selection:

Selected because of its proven suitability in toxicology studies and to comply with regulatory requirements for testing in a rodent animal species.

Sex:

male/female

Details on test animals or test system and environmental conditions:

Breeder: Charles River Laboratories Germany GmbH, Germany
Body weight (at 1st dosing): 227 - 329 g
Age of animals (at 1st dosing): 62 days
Adaptation period: 8 days
Diet: commercial diet ad libitum. Food residue was removed and weighed weekly.
Drinking water: ad libitum
Housing: singly in MAKROLON cages (type III plus)
Room temperature: 22 +/- 3 °C
Relative humidity: 55 +/- 10%
Light / Dark: 12 hrs / 12 hrs

Route of administration:

oral: gavage

Vehicle:

methylcellulose

Details on oral exposure:

Preparation of the test item-formulations: The test item was suspended in the vehicle to the appropriate concentrations. Test item formulations with concentrations of 10, 30 and 100 mg Famex LL T/mL were prepared. The test item formulations were freshly prepared every day.
ration
Vehicle / Control: 0.5% Methylcellulose 400
Administration volume: 10 mL/kg b.w./day
Selection of route of administration: According to international guidelines.
The control animals received the vehicle at the same administration volume in the same way.
The amount of the test item was adjusted to each animal's current body weight daily.

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

Test item formulation analysis: Tests by appropriate analytical methods were carried out for the pre-test study (LPT Study No. 34891) to determine the concentration, homogeneity and stability of the test item in the formulations. The results revealed that the test item formulations were correctly prepared by LPT and that the formulations were very homogeneous and stable for at least 24 hours.

Duration of treatment / exposure:

28 consecutive days

Frequency of treatment:

Once daily

Dose / conc.:

0 mg/kg bw/day (nominal)

Dose / conc.:

100 mg/kg bw/day (nominal)

Dose / conc.:

300 mg/kg bw/day (nominal)

Dose / conc.:

1 000 mg/kg bw/day (nominal)

No. of animals per sex per dose:

Number and sex of animals: 40 animals (20 male and 20 female rats)
5 animals/sex/group
4 spare animals (2 males and 2 females) of the same delivery were available for possible replacement during the adaptation period, however, not used

Control animals:

yes, concurrent vehicle

Details on study design:

Rationale for dose selection
The dose levels were selected in agreement with the Sponsor based on the results of a 7 day dose-range-finding study (LPT Study No. 34891). In this dose-range-finding study rats were treated with 100, 300 and 1000 mg Famex LL T/kg b.w./day by oral administration via gavage once daily for 7 days. None of the animals died or had to be sacrificed prematurely. No changes in behaviour, external appearance or faeces were noted. No test item-related influence was noted on the body weight and the food and drinking water consumption. No changes were noted at macroscopic inspection. After consideration of these data dose levels of 100, 300 and 1000 mg Famex LL T/kg b.w./day were selected for this 28 day toxicity study.

Positive control:

no

Observations and examinations performed and frequency:

Clinical signs: Each animal was observed before and after dosing at each time of dosing for any signs of behavioural changes, reaction to treatment, or illness. Animals were checked regularly throughout the working day from 7.30 a.m. to 4.30 p.m. On Saturdays and Sundays animals were checked regularly from 8.00 a.m. to 12.00 a.m. with a final check performed at approximately 4.00 p.m. Additionally, once before the first exposure (to allow for within-subject comparisons) and once a week thereafter (always on the first day of the test week), detailed clinical observations were made in all animals. In test week 4 these observations were performed prior to any laboratory investigations. These observations were made outside the home cage in a standard arena and at the same time of day, each time. Signs noted included changes in skin, fur, eyes, mucous membranes, occurrence of secretions and excretions and autonomic activity (e.g. lacrimation, pilo-erection, pupil size, unusual respiratory pattern). Changes in gait, posture and response to handling as well as the presence of clonic or tonic movements, stereotypies (e.g. excessive grooming, repetitive circling) or bizarre behaviour (e.g. self-mutilation, walking backwards) were also recorded.
Neurological screening: At the end of test week 4 (on test day 28), screening of sensory reactivity to stimuli of different types (e.g. auditory, visual and proprioceptive stimuli) (based on GAD ), as well as the assessment of grip strength (MEYER ) and motor activity assessment were conducted in all animals 2 hours after dosing outside the home cage.
Functional tests: Grip strength and Locomotor activity.
Mortality: Checks were made early in the morning and again in the afternoon of each working day to look for dead or moribund animals. This would have allowed post mortem examination to be carried out during the working period of that day. On Saturdays and Sundays a similar procedure was followed except that the final check was carried out at approximately 4:00 p.m. However, no premature deaths occurred in this study.
Body weight: The weight of each rat was recorded at the time of group allocation, on the day of commencement of treatment and daily thereafter for dose adjustment. Weekly values are stated in the report.
Food consumption: The quantity of food left by individual animals was recorded on a weekly basis throughout the experimental period. Food intake per rat (g/rat/week) was calculated using the total amount of food given to and left by each rat in each group upon completion of a treatment week. From these data the food consumption (in g/kg b.w./day) was
determined.
The drinking water consumption was monitored daily by visual appraisal throughout the study.
Laboratory examinations: Standard parameters concerning haematology, coagulation and clinical chemistry were recorded.
Ophthalmological and auditory examinations: Examinations were performed on all animals before first dosing and at study termination (on test day 28)

Sacrifice and pathology:

Necropsy: On test day 29 (approximately 24 hours after the last administration) the animals were dissected following a randomisation scheme. The animals were euthanized by carbon dioxide (CO2) inhalation, exsanguinated by cutting the aorta abdominalis, weighed, dissected and inspected macroscopically under the direction of a pathologist. All superficial tissues were examined visually and by palpation and the cranial roof was removed to allow observation of the brain, pituitary gland and cranial nerves. After ventral midline incision and skin reflection, all subcutaneous tissues were examined. The condition of the thoracic viscera was noted with due attention to the thymus, lymph nodes and heart. The abdominal viscera were examined before and after removal, the urinary bladder was examined externally and by palpation. The gastro-intestinal tract was examined as a whole and the stomach and caecum were incised and examined. The lungs were removed and all pleural surfaces examined under suitable illumination. The liver and the kidneys were examined. Any abnormalities in the appearance and size of the gonads, adrenal glands, uterus, intra-abdominal lymph nodes and accessory reproductive organs were recorded. The weights of the following organs of all animals were determined before fixation: Adrenal gland (2), Brain, Epididymis (2), Heart, Kidney (2), Liver, Ovary (2), Spleen, Testicle (2), Thymus, Thyroid incl. parathyroids (1), Uterus incl. cervix) and as whole: Prostate and seminal vesicles with coagulating glands.
Histopathology: Routinely examination of all relevant organs.

Statistics:

Standard methods for statistics were applied.

Clinical signs:

no effects observed

Mortality:

no mortality observed

Body weight and weight changes:

no effects observed

Food consumption and compound intake (if feeding study):

no effects observed

Food efficiency:

no effects observed

Water consumption and compound intake (if drinking water study):

no effects observed

Ophthalmological findings:

no effects observed

Haematological findings:

no effects observed

Clinical biochemistry findings:

no effects observed

Urinalysis findings:

no effects observed

Behaviour (functional findings):

no effects observed

Immunological findings:

not examined

Organ weight findings including organ / body weight ratios:

no effects observed

Gross pathological findings:

no effects observed

Neuropathological findings:

no effects observed

Histopathological findings: non-neoplastic:

no effects observed

Histopathological findings: neoplastic:

no effects observed

Key result

Dose descriptor:

NOEL

Effect level:

> 1 000 mg/kg bw/day (nominal)

Based on:

test mat.

Sex:

male/female

Remarks on result:

not determinable due to absence of adverse toxic effects

Key result

Critical effects observed:

no

Conclusions:

The aim of this repeated dose toxicity study was to obtain information on the toxicity of Famex LL T administered once daily by oral administration via gavage to rats for 28 consecutive days. The animals were treated with 100, 300 or 1000 mg Famex LL T/kg b.w./day. The control animals received the vehicle (0.5% Methylcellulose 400). None of the animals died or had to be sacrificed prematurely. No test item-related changes were observed for the behaviour or external appearance of the animals, the detailed clinical observations, the neurological screening, the body weight, body weight gain and body weight at autopsy, the food and drinking water consumption, for any of the haematological and clinical chemical parameters, the eyes or optic region, the auditory acuity, the relative and absolute organ weights, and at macroscopic inspection at necropsy at any dose level. The histopathological examination did not reveal any test item-related morphological changes.
The experimental no-observed-effect level (NOEL) was above 1000 mg Famex LL T/kg b.w./day, by daily oral administration.

Executive summary:

The aim of this repeated dose toxicity study was to obtain information on the toxicity of Famex LL T administered once daily by oral administration via gavage to rats for 28 consecutive days. The animals were treated with 100, 300 or 1000 mg Famex LL T/kg b.w./day. The control animals received the vehicle (0.5% Methylcellulose 400). None of the animals died or had to be sacrificed prematurely. No test item-related changes were observed for the behaviour or external appearance of the animals, the detailed clinical observations, the neurological screening, the body weight, body weight gain and body weight at autopsy, the food and drinking water consumption, for any of the haematological and clinical chemical parameters, the eyes or optic region, the auditory acuity, the relative and absolute organ weights, and at macroscopic inspection at necropsy at any dose level. The histopathological examination did not reveal any test item-related morphological changes.

The experimental no-observed-effect level (NOEL) was above 1000 mg Famex LL T/kg b.w./day, by daily oral administration.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

NOAEL

1 000 mg/kg bw/day

Study duration:

subacute

Species:

rat

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

Justification for classification or non-classification

The test item was tested in a 28 day study in rats with daily oral dosing of 0, 100, 300 and 1000 mg/kg via gavage. The experimental no-observed-effect level (NOEL) was above 1000 mg/kg. Therefore, there is no need for classification and labelling of the test item according to CLP Regulation 1272/2008/EG.