1,4-diazabicyclooctane

Toxicity to reproduction

Administrative data

Endpoint:

one-generation reproductive toxicity

Remarks:

based on generations indicated in Effect levels (migrated information)

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: This study was conducted according to OECD 422 guidelines and GLP principles

Cross-reference

Reason / purpose for cross-reference:

reference to same study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2000

Report date:

2000

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 422 (Combined Repeated Dose Toxicity Study with the Reproduction / Developmental Toxicity Screening Test)

GLP compliance:

yes

Limit test:

no

Test material

Test material information

Constituent 1

Reference substance name:

1,4-diazabicyclooctane

EC Number:

205-999-9

EC Name:

1,4-diazabicyclooctane

Cas Number:

280-57-9

Molecular formula:

C6H12N2

IUPAC Name:

1,4-diazabicyclooctane

Test animals

Species:

rat

Strain:

Sprague-Dawley

Sex:

male/female

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

water

Remarks:

deionized

Details on mating procedure:

Following 14 days of dose administration, 10 F0 males were paired 1:1 with 10 F0 females from the same treatment group in the Reproductive/Developmental Toxicity Phase on September 7, 1999. A breeding record containing the male and female identification numbers and the date of cohabitation was generated. Positive evidence of mating was confirmed by the presence of sperm in a vaginal smear or a copulatory plug. Each mating pair was examined daily with the following exception. On study day 25 (September 18, 1999), the examination for evidence of mating for 300 mg/kg/day group female no. 28451 was inadvertently not documented. This female was determined to be nongravid at the scheduled necropsy. This deviation had no impact on the integrity or outcome of the study due to its singular occurrence. The day when evidence of mating was identified was termed day 0 of gestation. A maximum of 14 days was allowed for mating. Following 14 days of cohabitation, females that did not have positive evidence of mating were transferred to plastic maternity cages containing nesting material. It should be noted that animals selected for the Recovery Phase (five rats/sex, Groups 1 and 4) were not to be evaluated for reproductive parameters. However, the Recovery Phase animals were inadvertently paired for breeding. The females that did not show evidence of mating by the presence of a copulatory plug or by a vaginal smear for sperm were randomized, via a computer program, to determine the five females assigned to the Recovery Phase. The males paired with these selected females were also assigned to the Recovery Phase. This deviation did not affect the quality or integrity of the study.

Duration of treatment / exposure:

Exposure period: 29 days
Premating exposure period (males): 14 days
Premating exposure period (females): 14 days
Duration of test: 57 days

Frequency of treatment:

daily

Doses / concentrations

Remarks:

Doses / Concentrations:
0, 100, 300, 1000 mg/kg/day
Basis:

Control animals:

yes, concurrent vehicle

Examinations

Litter observations:

All females from each dose group in the Reproduction/Developmental Phase were allowed to deliver naturally and rear their young to postnatal day 4 (PND 4). During the period of expected parturition, the females were observed twice daily for initiation and completion of parturition and for signs of dystocia. The duration of gestation was calculated for each animal using the date delivery initiated. The day on which delivery was judged complete was designated lactation day 0.

When parturition was complete, litters were sexed and examined for gross malformations, and the number of stillborn and live pups was recorded. All pups were individually identified by the application of tattoo markings on the digits at the completion of parturition. Each litter was observed daily for survival and all deaths were recorded. A daily record of litter size was maintained.

Litters were examined daily for any adverse changes in appearance and behavior. Each pup received a detailed clinical examination on PND 1 and 4. Any abnormalities in nesting and nursing behavior were recorded. Each pup was sexed on lactation days 0 and 4.

Pups were individually weighed on PND 1 and 4.

All surviving pups were euthanized and necropsied on PND 4 with emphasis placed on developmental morphology. Stillborn pups and pups dying between birth and PND 4 were also necropsied. Tissues were preserved in 10% neutral buffered formalin for possible future histopathologic examination only as deemed necessary by the gross findings. All carcasses were then discarded.

Results and discussion

Results: P0 (first parental generation)

General toxicity (P0)

Clinical signs:

effects observed, treatment-related

Reproductive function / performance (P0)

Reproductive performance:

no effects observed

Details on results (P0)

No test article-related effects on male reproductive performance were observed at any dose level. The male mating indices were l00%, l00%, 90.0% and 100% in the control, 100, 300, and 1000 mg/kg/day groups, respectively. The male fertility indices were l00%, 100%. 90.0% and 90.0% in these same respective groups. No statistically significant differences from the control group values were observed. The mean number of days between pairing and coitus in the treated groups was not adversely affected by test article administration. The mean number of days between pairing and coitus in the 1000 mg/kg/day group (2.0 days) was reduced and statistically significant (p<0.01) when compared to the control group value (3.1 days). The mean value for the pre-coital interval in the WIL historical control data is 3.1 days; the minimum value is 2.0 days. Therefore, this decrease in pre-coital interval was not attributed to test article administration.

Effect levels (P0)

Dose descriptor:

NOAEL

Effect level:

100 mg/kg bw/day

Based on:

test mat.

Sex:

female

Basis for effect level:

clinical signs

mortality

Results: F1 generation

Effect levels (F1)

Dose descriptor:

NOAEL

Generation:

F1

Effect level:

300 mg/kg bw/day

Based on:

test mat.

Sex:

male/female

Basis for effect level:

clinical signs

mortality

Overall reproductive toxicity

Reproductive effects observed:

not specified

Any other information on results incl. tables

Oral administration of Triethylenediamine at dose levels of 100, 300, and 1000 mg/kg/day did not adversely affect F0 survival, pregnancy status or duration of gestation.

 

Test article-related effects on reproductive performance were limited to the 1000 mg/kg/day group and consisted of a slight decrease in the mean number of pups born per litter and the live litter size on PND 0. One female in the 1000 mg/kg/day group had an entirely resorbed litter. The only potential disturbance in parturition was the euthanasia of a single 1000 mg/kg/day group female on lactation Day 0 due to dystocia. This female delivered nine pups, had three late resorptions retained in utero and one late resorption retained in the vagina. No other test article-related effects were noted on male and female reproductive performances at any dose level.

Applicant's summary and conclusion

Conclusions:

Oral administration of Triethylenediamine resulted in F0 toxicity in both males and females at a dose level of 1000 mg/kg/day as evidenced by changes in clinical conditions of the animals, reduced body weight and food consumption, reduced motor activity (females only), increased serum alkaline phosphatase concentrations (females only), increased liver weights (females only) and microscopic changes (inflammatory and/or proliferative lesions) in the kidneys and urinary bladder of a single 1000 mg/kg/day group female, none of the above findings persisted to the end of the 14-day recovery period. F0 toxicity in the 300 mg/kg/day groups was limited to chronic inflammation of the kidneys in the males. There were no indications of F0 toxicity in the 100 mg/kg/day group males and females. 
 
F0 mating and fertility indices were not affected by test article administration.
 
F1 neonatal toxicity was exhibited at 1000 mg/kg/day by decreased live litter size, decreased postnatal pup survival and decreased pup body weights. No indications of neonatal toxicity were observed at 100 and 300 mg/kg/day.
 
Based on the data obtained, NOAEL (no-observed-adverse-effect level) for F0 reproductive toxicity was considered to be 300 mg/kg/day. The NOAEL for Fl neonatal toxicity was considered to be 300 mg/kg/day. The NOAEL for F0 parental systemic toxicity was considered to be 100 mg/kg/day.

Executive summary:

Oral administration of Triethylenediamine resulted in F0 toxicity in both males and females at a dose level of 1000 mg/kg/day as evidenced by changes in clinical conditions of the animals, reduced body weight and food consumption, reduced motor activity (females only), increased serum alkaline phosphatase concentrations (females only), increased liver weights (females only) and microscopic changes (inflammatory and/or proliferative lesions) in the kidneys and urinary bladder of a single 1000 mg/kg/day group female, none of the above findings persisted to the end of the 14-day recovery period. F0 toxicity in the 300 mg/kg/day groups was limited to chronic inflammation of the kidneys in the males. There were no indications of F0 toxicity in the 100 mg/kg/day group males and females. 

 

F0 mating and fertility indices were not affected by test article administration.

 

F1 neonatal toxicity was exhibited at 1000 mg/kg/day by decreased live litter size, decreased postnatal pup survival and decreased pup body weights. No indications of neonatal toxicity were observed at 100 and 300 mg/kg/day.

 

Based on the data obtained, NOAEL (no-observed-adverse-effect level) for F0 reproductive toxicity was considered to be 300 mg/kg/day. The NOAEL for Fl neonatal toxicity was considered to be 300 mg/kg/day. The NOAEL for F0 parental systemic toxicity was considered to be 100 mg/kg/day.