Registration Dossier

Toxicological information

Repeated dose toxicity: oral

Currently viewing:

Administrative data

Endpoint:
short-term repeated dose toxicity: oral
Type of information:
experimental study
Adequacy of study:
supporting study
Study period:
2015-08-31 to 2015-10-02
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Preliminary toxicity study, with supporting results, not according to OECD and EC guidelines but based on OECD 407 and GLP principles.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2015
Report Date:
2015

Materials and methods

Principles of method if other than guideline:
The preliminary toxicity study was performed based on the principles described in OECD guideline 407.
GLP compliance:
no
Remarks:
The work was performed following GLP principles, no claim is made.
Limit test:
no

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
liquid
Details on test material:
- Physical state: liquid
- Storage condition of test material: Room temperature
Specific details on test material used for the study:
- Name as cited in study report: POPDA, reaction products of propane-1,2-diol, propoxylated by animation of the terminal hydroxyl groups
- Color: yellow
- Analytical purity: 100%
- Impurities: water 0.12%
- Lot/Batch number: DR66700414
- Expiry date: 2016-12-05
- Stability: satisfactory stability was documented at 10 and 200 mg/ml following refrigerated (nominally 4-8°C) or ambient (nominally 21°C) storage for 15 days
- Storage: dark
- Total correction factor: none
-Ppurity/weighing factor: none

Test animals

Species:
rabbit
Strain:
New Zealand White
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: 6 female New Zealand White rabbits, obtained from stock
- Age at start of study (day 1): 12 to 40 weeks old
- Weight at start of study (day 1): 3.00 to 3.44 kg
- Fasting period before study: no data
- Housing: individually housed in suspended plastic cages fitted with perforated floor panels and mounted in batteries. Undertrays lined with absorbent paper were changed at least 3 times a week.
- Diet (e.g. ad libitum): restricted availability (200 g/animal/day), Harlan High Fiber 2931C diet
- Water (e.g. ad libitum): ad libitum, potable water from the public supply via polycarbonate bottles with sipper tubes
- Acclimation period: 3 days before commencement of treatment

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 16-20 °C
- Humidity (%): 40-70%
- Air changes (per hr): filtered fresh air which was passed to atmosphere and not recirculated, no further data
- Photoperiod (hrs dark / hrs light): 12/12, artificial lighting

IN-LIFE DATES:
- group 1: from 2015-09-03 to 2015-09-12
- group 2: from 2015-09-21 to 2015-10-02

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
water
Remarks:
purified
Details on oral exposure:
PREPARATION OF DOSING SOLUTIONS:
- An appropriate volume of vehicle was added to the test substance and mixed by magnetic stirring until dissolution was achieved. A series of solutions at the required concentrations were prepared by serial dilution of the primary solution with the vehicle. Formulations were stirred using a magnetic stirrer before and throughout the dosing procedure.
- Frequency of preparation: weekly
- Storage of formulation: ambient temperature (nominally 21°C)
- Detailed records of compound usage were maintained. The amount of test substance necessary to prepare the formulations and the amount actually used were determined on each occasion. The difference between these amounts was checked before the formulations were dispensed.


VEHICLE
- Concentration in vehicle: 60 mg/ml (300 mg/kg) and 120 mg/ml (600 mg/kg)
- Amount of vehicle (if gavage): 5 ml/kg
Analytical verification of doses or concentrations:
no
Duration of treatment / exposure:
group 1: 10 days (group prematurely terminated on day 10 of study)
group 2: 11 days (group prematurely terminated on day 12 of study, with animals having received 11 consecutive daily doses)
Frequency of treatment:
group 1: single doses on days 1 and 8 - third planned dose on day 15 was not administered
group 2: repeated daily dosing (11 occasions)
Doses / concentrationsopen allclose all
Dose / conc.:
300 mg/kg bw/day (actual dose received)
Remarks:
group 1 day 1, group 2
Dose / conc.:
600 mg/kg bw/day (actual dose received)
Remarks:
group 1, day 8
No. of animals per sex per dose:
3 females/group; 2 groups
Control animals:
no
Details on study design:
- Dose selection rationale: the starting dose of 300 mg/kg/day was selected by the sponsor. The subsequent dose levels for group 1 was based upon the apparent lack of a reaction observed, following a single dose at 300 mg/kg/day. Due to marked effects in the stomach of group 1 animals following a single dose at 600 mg/kg/day, the dose level for group 2 animals for repeated daily dosing was set at 300 mg/kg/day.
Positive control:
no

Examinations

Observations and examinations performed and frequency:
CAGE SIDE OBSERVATIONS: Yes
- Time schedule: at least twice daily (at least once daily during acclimatization period)
- Animals were inspected visually for evidence of ill-health or reaction to treatment. Cages were inspected for evidence of animal ill-health amongst the occupant.

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: at least twice daily:
- on dosing days: predose observation, upon completion of dosing, 30 minutes to 1 hour after completion of dosing, 2 to 3 hours after completion of dosing, as late as possible in the working day;
-on non-dosing days: in the morning, as late as possible in the working day.

BODY WEIGHT: Yes
- Time schedule for examinations: weekly during acclimatization, daily from 3 days prior to the start of treatment (Day -3), on the day that treatment commenced (Day 1), daily throughout the study and before necropsy.

FOOD CONSUMPTION:
- The weight of food supplied to each cage, that remaining and an estimate of any spilled was recorded daily from 3 days prior to the start of treatment (day -3) and throughout the study.

FOOD EFFICIENCY: No

WATER CONSUMPTION AND COMPOUND INTAKE (if drinking water study): No

OPHTHALMOSCOPIC EXAMINATION: No

HAEMATOLOGY: No

CLINICAL CHEMISTRY: No

URINALYSIS: No

NEUROBEHAVIOURAL EXAMINATION: No

OTHER:
MORTALITY:
- A viability check was performed near the start and end of each working day.
Sacrifice and pathology:
GROSS PATHOLOGY: Yes
- All animals were subject to a detailed necropsy. After a review of the history of each animal, a full macroscopic examination of the tissues was performed. All external features and orifices were examined visually. Any abnormality in the appearance or size of any organ and tissue (external and cut surface) was recorded and the required tissue samples preserved in appropriate fixative. In light of the macropathology findings for the Group 1 decedent animal, the stomach was also retained.
- The retained tissues were checked before disposal of the carcass.
- Schedule: Group 1 animals: day 10 (2 days after last dose); Group 2 animals: day 12 (1 day after last dose)
- Method of kill: Intravenous injection of sodium pentobarbitone.

HISTOPATHOLOGY: No
Statistics:
No statitical analysis performed.

Results and discussion

Results of examinations

Clinical signs:
effects observed, treatment-related
Description (incidence and severity):
- Following a single dose at 300 mg/kg/day in group 1, there were no marked clinical signs to Day 8 of study. A single dose of PODPA at 300 mg/kg/day was well tolerated.
- Following a single dose at 600 mg/kg in group 1, one female (number 2) showed grinding of teeth the day after dosing (day 9). On Day 10 of study (2 days after dosing at 600 mg/kg/day) female 2 was found dead.
- Repeat dosing of 300 mg/kg/day (group 2) elicited clinical signs of little hay eaten and few faeces for the two most affected females (numbers 4 and 6). Clinical signs of female 5 were restricted to little hay eaten on one occasion.
Mortality:
mortality observed, treatment-related
Description (incidence):
- Female 4 (group 2) was found dead on day 12, having received 11 consecutive daily doses at 300 mg/kg/day.
Body weight and weight changes:
effects observed, treatment-related
Description (incidence and severity):
- Following a single dose at 300 mg/kg/day in group 1, there were no adverse effects upon body weight performance to Day 8 of study. A single dose of PODPA at 300 mg/kg/day was well tolerated.
- Repeat dosing of 300 mg/kg/day (group 2) elicited a marked reduction in food intake in 2 out of 3 females (number 4 and 6), with subsequent body weight loss of between 210 and 220 g. Female 5 showed no clear or marked effect of treatment upon food consumption. Female 5 and overall body weight loss of just 20 g was recorded. The reduction in food intake was considered treatment related, with secondary effects manifest as body weight loss and clinical signs consistent with inappetance.
Food consumption and compound intake (if feeding study):
not examined
Description (incidence and severity):
- Following a single dose at 300 mg/kg/day in group 1, there were no adverse effects upon food consumption to Day 8 of study. A single dose of PODPA at 300 mg/kg/day was well tolerated.
- Following a single dose at a dose level of 600 mg/kg in group 1, all animals showed reduced food intake from the day of dosing.
- Repeat dosing of 300 mg/kg/day (group 2) elicited a marked reduction in food intake in 2 out of 3 females (number 4 and 6), with subsequent body weight loss of between 210 and 220 g. Female 5 showed no clear or marked effect of treatment upon food consumption. Female 5 and overall body weight loss of just 20 g was recorded. The reduction in food intake was considered treatment related, with secondary effects manifest as body weight loss and clinical signs consistent with inappetance.

Food efficiency:
not examined
Water consumption and compound intake (if drinking water study):
not examined
Ophthalmological findings:
not examined
Haematological findings:
not examined
Clinical biochemistry findings:
not examined
Urinalysis findings:
not examined
Behaviour (functional findings):
not examined
Organ weight findings including organ / body weight ratios:
not examined
Gross pathological findings:
effects observed, treatment-related
Description (incidence and severity):
- Macroscopic examination of female 2 (group 1) revealed red fluid in the abdomen, thickened omental adipose tissue with poorly defined dark areas, dark areas on the jejunum which contained yellow viscous fluid, a pale liver, and a pyloric perforation and several depression(s) in the pylorus of the stomach.
- Macroscopic examination of females 1 and 3 revealed findings including pale liver and/or duodenum, depressions on the pylorus and corpus, thickened pylorus, a distended stomach with liquefied or clear gelatinous contents and yellow viscous fluid in the jejunum. These findings were considered treatment-related.
- Macroscopic examination of female 4 (group 2) revealed dark areas on the mucosal aspect of the caecum, which contained liquified contents, multiple pylorus depressions on the stomach which was also distended and contained clear, viscous fluid and a thickened pylorus, and a depression of the stomach fundus. Due to the similarity of the findings of the GI tract in this decedent to those observed at 600 mg/kg/day, treatment at 300 mg/kg/day was subsequently suspended with surviving females killed for reasons of animal welfare on day 12 of their treatment period, having received 11 consecutive daily doses.
- Macroscopic examination of female 6 revealed a distended caecum, with liquified contents, a pale liver, multiple pylorus depressions and dark stomach contents.
- There were no macroscopic findings observed in female 5.
- The consistent macroscopic findings relating to the GI tract observed in two out of three females following repeat daily dosing at 300 mg/kg/day were considered treatment related.
Histopathological findings: non-neoplastic:
not examined
Histopathological findings: neoplastic:
not examined
Description (incidence and severity):
- Due to the marked macroscopic findings in female 2 of group 1, which were considered treatment related, and the reduced food intake and body weight loss observed in surviving females, which was also considered treatment related, with the associated clinical sign of little hay eaten, this dose group (group 1) was terminated and a dose of 600 mg/kg/day was considered unsuitable for any further investigation. Surviving females 1 and 3 were subsequently killed for reasons of animal welfare.

Target system / organ toxicity

Critical effects observed:
not specified

Applicant's summary and conclusion

Conclusions:
Dose levels of 300 mg/kg/day or above are clearly unsuitable for any further repeat dosing investigations in the female New Zealand White Rabbit.
Due to the marked nature of the macroscopic necropsy findings observed in the stomach and gastrointestinal tract, a recommended high dose level for the first phase of a two-phased preliminary embryo-fetal study in the New Zealand White Rabbit would be 150 mg/kg/day.