2-Butyne-1,4-diol, polymer with 2-(chloromethyl)oxirane, brominated, dehydrochlorinated, methoxylated

Repeated dose toxicity: oral

Administrative data

Endpoint:

short-term repeated dose toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP compliant, non-guideline study, available as unpublished report, no restrictions, fully adequate for assessment.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2014

Materials and methods

Test guideline

Qualifier:

equivalent or similar to guideline

Guideline:

other: OECD Guidelines for the Testing of Chemicals (425), "Acute Oral Toxicity: Up and Down Procedure," adopted: 3 October, 2008

GLP compliance:

yes (incl. QA statement)

Limit test:

yes

Test material

Test material information

Constituent 1

Reference substance name:

2-Butyne-1,4-diol, polymer with 2-(chloromethyl)oxirane, brominated, dehydrochlorinated, methoxylated

EC Number:

614-503-3

Cas Number:

68441-62-3

Molecular formula:

(C3H7O2)xC4H4O2Br2(C4H9O2)y with x + y = 2.5

IUPAC Name:

2-Butyne-1,4-diol, polymer with 2-(chloromethyl)oxirane, brominated, dehydrochlorinated, methoxylated

Test material form:

liquid: viscous

Details on test material:

- Name of test material (as cited in study report): Polyol IXOL B 350
- Substance type: Halogenated polyetherpolyol
- Physical state: viscous, dark brown liquid
- Analytical purity: ≥99%
- Lot/batch No.: 08-01-2013
- Expiration date of the lot/batch: 2015-01-08
- Storage condition of test material: ambient temperature, protected from light

Test animals

Species:

rat

Strain:

Sprague-Dawley

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: BioLASCO Taiwan Co., Ltd.
- Age at study initiation: Females and males, approx. 7 weeks,
- Weight at study initiation: 181 to 216 g (females) and 260 to 290 g (males),
- Housing: On arrival, animals were individually housed until randomization. Following randomization, animals were group housed (up to 3 animals of the same sex and same dosing group together) in solid bottom cages, with corn cob bedding
- Diet: rodent feed, ad libitum
- Water: reverse-osmosis purified and chlorinated water, ad libitum
- Acclimation period: The pretest period (from arrival to the study to first day of dosing) was 14 days.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): between 21.7°C to 23°C,
- Humidity (%): between 40.1% to 73.9%
- Air changes (per hr): 10 to 20
- Photoperiod (hrs dark / hrs light): 12/12

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

other: (w/v) Gum tragacanth powder in purified water

Details on oral exposure:

PREPARATION OF DOSING SOLUTIONS:
Formulations (w/w) were prepared at least once weekly and were stirred until a homogeneous suspension was achieved. The dose formulations were stirred at room temperature for at least 30 minutes before dosing and continuously during dosing. No correction was made for the purity of the test substance. Solutions were stored at ambient temperature.

VEHICLE
- Justification for use and choice of vehicle: 1% (w/v) Gum tragacanth powder in purified water. This vehicle was selected based on trial formulations performed at WuXi AppTec.
- Amount of vehicle (if gavage): 10 mL/kg body weight. Actual dose volumes were calculated according to the latest body weight.

Details on analytical verification of doses or concentrations:

CONCENTRATION VERIFICATION:
The concentration of Polyol IXOL B350 was determined in all dosing formulations according to a validated method (WuXi AppTec Study No.: 239-0002-AC).

Duration of treatment / exposure:

14 days.

Frequency of treatment:

Once daily

Doses / concentrations

Doses / concentrations 1

Dose / conc.:

417 mg/kg bw/day (nominal)

Doses / concentrations 2

Dose / conc.:

625 mg/kg bw/day (nominal)

Doses / concentrations 3

Dose / conc.:

938 mg/kg bw/day (nominal)

No. of animals per sex per dose:

16/sex per dose

Control animals:

yes, concurrent vehicle

Details on study design:

The purpose of this study was to assess the maximum tolerable dose of Polyol IXOL B350 when administered by oral gavage for 14 days to rats. Dosages were selected based on the data from the previous acute study (Koopman, 1985). Single oral dose at 1250 mg/kg caused mortality (3/5) and signs noted included but not limited to decreased respiration rate, decreased locomotor activity, abnormal posture and gait, pilo-erection, abnormal touch response, and hypothermia. Onset of most signs was between 6 and 24 hours post dose. Signs disappeared 48 hours post dose. No signs were observed after a single oral dose at 625 mg/kg. Therefore, 938 mg/kg/day was selected as the highest dose level for this study.

Examinations

Observations and examinations performed and frequency:

VIABILITY:
Viability (morbidity and mortality) checks were made twice daily, except on animal arrival and the day of necropsy, where animals were examined at least once.

CAGE SIDE OBSERVATIONS: Yes
- Time schedule: Daily during prestudy from Day -2, twice daily during the dosing (at approximately 2 to 3 hours and 10 to 12 hours postdose)
phase.

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: Once during prestudy, once predose on Day 1 for all study animals, and once weekly thereafter during the dosing (at
approximately 4 to 6 hours postdose) phase. Detailed observations were also conducted once on the day of scheduled necropsy.

BODY WEIGHT: Yes
- Time schedule for examinations: Once during pretest, on Day 1 prior to dosing, twice weekly throughout the treatment phase, and on the day of scheduled necropsy (fasted terminal body weight).

FOOD CONSUMPTION AND COMPOUND INTAKE
- Time schedule for examinations: Food consumption was measured (weighed) for all Study animals for once daily after randomization throughout the study (except during fasting periods). Animals were fasted overnight prior to scheduled necropsies and blood collections for clinical pathology.

CLINICAL PATHOLOGY
- All study animals were evaluated for hematology, clinical chemistry, and coagulation once prior to necropsy.

Sacrifice and pathology:

- Gross examination: A complete necropsy will be conducted on all study animals. At the end of the exposure period, surviving animals were anesthetized by isoflurane, euthanatized by exsanguination, and necropsied. A thorough autopsy was also performed on the animals that died intercurrently. The following organs of all surviving animals were weighed (paired organs together) as soon as possible after dissection to avoid drying: kidneys, liver, ovaries and testes
- Preservation: The tissues and organs of the animals of the range finding study mentioned above were preserved in a neutral aqueous phosphate-buffered 4 per cent solution of formaldehyde (10% solution of formalin).
- Histopathological examination: Histopathological examination was not performed.

Statistics:

All reported numerical data were subjected to calculations of group means and standard deviations, unless otherwise stated hereafter. Males and females were analyzed separately. Whenever there were more than two groups, the homogeneity of the group variances was evaluated using the Levene’s test at the 0.05 significance level.
Clinical observations and necropsy observations were organized into incidence tables to summarize findings. The data were evaluated for compound-related effects by making comparisons of individual and group values between the compound-treated and control groups, and between intervals where appropriate

Results and discussion

Results of examinations

Clinical signs:

no effects observed

Description (incidence and severity):

There were no test article-related findings observed during the 14 day dosing period.

Mortality:

no mortality observed

Description (incidence):

No mortality occurred during the study

Body weight and weight changes:

no effects observed

Description (incidence and severity):

There were no test article-related changes in body weight or body weight gain in males or females given Polyol IXOL B350 up to 938 mg/kg/day.

Food consumption and compound intake (if feeding study):

no effects observed

Description (incidence and severity):

There were no test article-related changes in food consumption.

Food efficiency:

not examined

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

not examined

Ophthalmological findings:

not examined

Haematological findings:

no effects observed

Description (incidence and severity):

No test article-related effects on hematology or coagulation at any dose levels tested.

Clinical biochemistry findings:

no effects observed

Description (incidence and severity):

No test article-related effects on serum chemistry at any dose levels tested.

Endocrine findings:

not examined

Urinalysis findings:

not examined

Behaviour (functional findings):

not examined

Immunological findings:

not examined

Organ weight findings including organ / body weight ratios:

effects observed, treatment-related

Description (incidence and severity):

Test article-related organ weight changes were noted in the livers from the rats given ≥417 mg/kg/day. Compared with the control group, statistically significant increases in the liver weights (absolute and relative to body weight and brain weight) were noted in both males (↑22% to 41%) and females (↑11% to 33%) given ≥417 mg/kg/day, which correlated with macroscopically noted enlarged liver in the males given ≥417 mg/kg/day.
Statistically significant increases in the kidney weights (absolute and relative to body weight) were noted in the males given 417 mg/kg/day and 938 mg/kg/day, these changes were not like test article-related due to lack of dose dependency.

Gross pathological findings:

effects observed, treatment-related

Description (incidence and severity):

Minimal/mild enlarged liver was noted in 2/4 males given 417 mg/kg/day, 3/4 males given 625 mg/kg/day and all 4 males given 938 mg/kg/day, which correlated with increased liver weights. Minimal enlarged thyroid glands were noted in 4 males given ≥417 mg/kg/day and 3 females given ≥625 mg/kg/day. Other changes were not considered as test article-related.

Neuropathological findings:

not examined

Histopathological findings: non-neoplastic:

not examined

Histopathological findings: neoplastic:

not examined

Effect levels

Dose descriptor:

LOAEL

Effect level:

417 mg/kg bw/day (actual dose received)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: Increased liver weights

Target system / organ toxicity

Critical effects observed:

not specified

Applicant's summary and conclusion

Conclusions:

A LOAEL of 417 mg/kg bw/day was established in a 14-day oral range finding study.

Executive summary:

Polyol IXOL B350 was tested in a GLP 14-day dose range finding study in male and female rat with application via the oral route (Hongping Wan, 2014). The dose range finding study was performed as preparation for the reproduction toxicity studies.

The test substance was administered as a solution in 1% (w/v) Gum tragacanth powder in purified water by oral gavage to 4 groups of 16 male and 16 female Sprague-Dawley rats each, once a day. The control group was administered vehicle only for 14 consecutive days. The test article-treated groups were dosed at 417, 625, or 938 mg/kg/day for 14 consecutive days. The dosing volume for all animals was 10 mL/kg. Animals were necropsied on Day 15. Toxicological assessments included morbidity/mortality, clinical observations, body weights, food consumption, clinical pathology (hematology, coagulation, and serum chemistry), organ weight and gross necropsy examination.

There were no test article-related effects on clinical signs, body weight, food consumption, hematology, coagulation, or serum chemistry at any dose levels tested. Test article-related pathology changes were noted as enlarged liver in the males with increased liver weights in both sexes given ≥417 mg/kg/day, and enlarged thyroid glands in the males given ≥417 mg/kg/day and females given ≥625 mg/kg/day. Based on the increased liver weights (females and males at all dose levels) and enlarged thyroid glands in males (males at all dose levels, females at mid and high dose), a LOAEL of 417 mg/kg bw/day was established.