N,N'''-1,6-hexanediylbis[N'-cyanoguanidine]

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)

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1993

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

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

GLP compliance:

yes

Limit test:

no

Test material

Test material information

Constituent 1

Test material form:

solid: crystalline

Details on test material:

The dose preparations were corrected for purity (68.9%)

Test animals

Species:

rat

Strain:

Wistar

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Barriered Animal Breeding Unit, Zeneca
- Age at study initiation: 5 weeks
- Weight at study initiation: mean weight = 190g males, 160g females
- Housing: stainless steel cages 26.5 x 50.0 x 20.7 cm, 5 animals per cage
- Diet (e.g. ad libitum): ad libitum
- Water (e.g. ad libitum): ad libitum
- Acclimation period: 1 week

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19-23°C
- Humidity (%):40-70%
- Air changes (per hr): 25-30
- Photoperiod (hrs dark / hrs light): 12/12

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

other: 0.5% (w/v) hydroxypropylmethylcellulose in 0.1% (w/v) aqueous polysorbate 80

Details on oral exposure:

Dosing formulations were shaken then stirred using a magnetic stirrer prior to and during dosing. Rats were dosed by gavage at 1.0ml/100g according to their daily individual bodyweights. Rats were dosed sequentially within group order at approximately the same time each day.

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

Portions of sample were dissolved in 25ml DMF and further diluted with methanol to give concentrations of approximately 40ug/ml HMBDA. Solutions were analysed by HPLC using a spherisorb S50DS1 column, mobile phase water/methanol/ammonium acetate (700:300:5 g/l), UV detection at 220nm. The sample concentrations were calcualted by reference to the interspersed calibration standard.

Duration of treatment / exposure:

Single oral gavage dose

Frequency of treatment:

Daily dosing for 28 consecutive days

Doses / concentrations

Doses / concentrations 1

Remarks:

Doses / Concentrations:
50 mg/kg bw/day
Basis:
actual ingested

Doses / concentrations 2

Remarks:

Doses / Concentrations:
250 mg/kg bw/day
Basis:
actual ingested

Doses / concentrations 3

Remarks:

Doses / Concentrations:
1000 mg/kg bw/day
Basis:
actual ingested

No. of animals per sex per dose:

5 males and 5 females

Control animals:

yes, concurrent vehicle

Details on study design:

Dose levels for the study were selected on the basis of a preliminary sighting study.

Positive control:

None

Examinations

Observations and examinations performed and frequency:

CAGE SIDE OBSERVATIONS: Yes
- Time schedule: Daily

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: Daily

BODY WEIGHT: Yes
- Time schedule for examinations: Daily

FOOD CONSUMPTION: Yes
- Food consumption for each group of animals wasa recorded continuously and calculated on a weekly basis

HAEMATOLOGY: Yes
- Time schedule for collection of blood: Termination
- Method: Cardiac puncture
- Parameters: haemoglobin, total white cell count, red cell count, mean cell colume, mean cell haemoglobin concentration, haematocrit, platelet count, prothrombin and kaolin-cephalin times, differential white cell count, red cell morphology

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: Termination
- Method: Cardiac puncture
- Parameters: plasma urea, creatinine, glucose, albumin, total protein, cholesterol, triglycerides, total bilirubin, sodium, potassium, chloride, calcium, phosphorous, plasma alkaline phosphatase, alanine transaminase, creatine kinase, aspartate transminase, gamma-glutamyl transferase

Sacrifice and pathology:

GROSS PATHOLOGY: Yes
- Organs weighed: Testes, adrenal glands, kidneys, liver

HISTOPATHOLOGY: Yes
- Organs examined by light microscopy: Adrenal glands, epididymis, heart, kidney, liver, spleen, testis, macroscopically abnormal tissue

Statistics:

Body weights were considered by analysis of covariance.
Haematology and blood clincial chemistry were considered by analysis of variance.
Organ weights were considered by analysis of variance and analysis of covariance on final bodyweight.

Results and discussion

Results of examinations

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:

not examined

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

not examined

Ophthalmological findings:

not examined

Haematological findings:

no effects observed

Clinical biochemistry findings:

no effects observed

Urinalysis findings:

not examined

Behaviour (functional findings):

not examined

Organ weight findings including organ / body weight ratios:

no effects observed

Gross pathological findings:

no effects observed

Histopathological findings: non-neoplastic:

no effects observed

Histopathological findings: neoplastic:

no effects observed

Effect levels

Dose descriptor:

NOEL

Effect level:

1 000 mg/kg bw/day (nominal)

Based on:

test mat.

Sex:

male/female

Target system / organ toxicity

Critical effects observed:

not specified

Any other information on results incl. tables

The achieved concentrations of test substance in vehicle were within 6% of nominal values. The homogeniety and stability of the dose formulation were shown to be satisfactory.

Applicant's summary and conclusion

Conclusions:

In a subacute oral study in the rat there were no effects of toxicological significance. The NOEL is 1000 mg/kg bw/day.