COAGULANT 122 (SOLID)

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

Study period:

2012-03-19 to 2012-10-26

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2012

Report date:

2012

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 421 (Reproduction / Developmental Toxicity Screening Test)

Deviations:

yes

Remarks:

The formulations for the stability trial were therefore prepared using purified water, and not distilled water as stated in the final protocol.

Principles of method if other than guideline:

The following deviations from protocol occurred:
Section 3.2 of the protocol stated that the vehicle on this study would be distilled water. At formulation preparation for the stability trial, study management
confirmed that purified water was considered equally acceptable for formulation preparation. The formulations for the stability trial were therefore prepared using purified water, and not distilled water as stated in the final protocol. The formulations for administration were prepared using purified water, as documented in Protocol Amendment 1.
Following the issue of Protocol Amendment 1, it was noted that distilled water was also referred to in the Method of Preparation in Section 3.2 of the protocol, but had not been altered in the amendment in error. A File Note acknowledged that the reference to distilled water remains in this section, but purified water was used for the formulations and as vehicle for the current study.
Sections 3.2 and 3.3 of the protocol refer to the establishment of homogeneity and stability as part of this study. The test item has been supplied as a solution, and the appropriate concentrations for use on the study had been and continued to be achieved by dilution of this solution. Establishment of homogeneity is not required for solutions, therefore only stability of the test item in the liquid matrix has been established and documented in Protocol Amendment 1.
These deviations were considered to have not affected the integrity of the study.

GLP compliance:

yes (incl. QA statement)

Limit test:

no

Test material

Test material information

Constituent 1

Test material form:

other: in aqueous solution

Details on test material:

Name of test material (as cited in study report): Coagulant 122 solution
Substance type: Colourless to yellow liquid
Purity: Supplied as a 30.8% solution
Batch number: 8351126521
Storage condition of test material: Room temperature (ca. 20°C in the dark)

Test animals

Species:

rat

Strain:

Crj: CD(SD)

Remarks:

Crl: CD(SD)

Sex:

male/female

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

water

Details on mating procedure:

Pairing to commence: After minimum of 2 weeks of treatment.

Male/female ration: 1:1

Duration of pairing: Up to 2 weeks.

Daily checks for evidence of mating: Ejected copulation plugs. Sperm within vaginal smear.

Day 0 of gestation: When positive evidence of mating detected.

Male/female separation: Day when mating evidence detected.

Pre-coital interval: Calculated for each female as time between first pairing and evidence of mating.

Analytical verification of doses or concentrations:

yes

Duration of treatment / exposure:

F0 animals For a minimum of 15 days before pairing until Day 6 after birth of F1 generation.
F1 animals No direct treatment

Frequency of treatment:

Once daily at approximately the same time each day.

Details on study schedule:

Study initiation: 19-Mar-12
(Protocol signed by Study Director)

Experimental start date: 26-Mar-12
(Pretreatment chemistry assays)

Animal arrival: 11-Apr-12
Treatment commenced: 16-Apr-12
Pairing commenced: 30-Apr-12
Necropsy completed: 3-Jun-12
Experimental completion date: 11-Jul-12
(Pathology)

Study completion: 26-Oct-12

Doses / concentrations

Doses / concentrations 1

Dose / conc.:

100 mg/kg bw/day (nominal)

Doses / concentrations 2

Dose / conc.:

300 mg/kg bw/day (nominal)

Doses / concentrations 3

Dose / conc.:

1 000 mg/kg bw/day (nominal)

No. of animals per sex per dose:

Group Treatment Dose (mg/kg/day) Number of animals Animal numbers
Male Female Male Female
1 Control 0 10 10 1-10 41-50
2 COAG-122 solution 100 10 10 11-20 51-60
3 COAG-122 solution 300 10 10 21-30 61-70
4 COAG-122 solution 1000 10 10 31-40 71-80

Control animals:

yes, concurrent vehicle

Results and discussion

Results: P0 (first parental generation)

General toxicity (P0)

Clinical signs:

no effects observed

Mortality:

mortality observed, non-treatment-related

Description (incidence):

One female of the high dose group, animal 79, was killed on Day 3 of lactation for welfare reasons because of poor litter condition/pup deaths and termination of the litter. At necropsy the mammary tissue appeared pale and inactive, although histological examination indicated the presence of secretory activity. The animal also had pelvic dilatation of the right kidney but this is a common background finding. There were no treatment-related findings in this animal.

Effect levels (P0)

Dose descriptor:

NOAEL

Effect level:

> 1 000 mg/kg bw/day

Based on:

not specified

Sex:

male/female

Basis for effect level:

reproductive performance

Remarks on result:

other:

Remarks:

Adult animals dosed with COAG-122 at doses of up to 1000 mg/kg/day showed minimal evidence of adverse response. During the first week of treatment effects were seen on male bodyweight at the high dose level but the animals quickly recovered to show normal growth patterns through the rest of the study. Reversible effects in the first week are not considered to equate to an "adverse" response. It was concluded that the No-Observed-Adverse-Effect-Level (NOAEL) for COAG-122 was in excess of 1000 mg/kg/day (the limit dose) for reproductive performance and offspring growth and survival in the CD rat following oral gavage administration in a standard screening test.

Results: P1 (second parental generation)

Effect levels (P1)

Dose descriptor:

NOAEL

Effect level:

> 1 000 mg/kg bw/day

Based on:

not specified

Sex:

male/female

Basis for effect level:

reproductive performance

Remarks on result:

other:

Remarks:

Adult animals dosed with COAG-122 at doses of up to 1000 mg/kg/day showed minimal evidence of adverse response. During the first week of treatment effects were seen on male bodyweight at the high dose level but the animals quickly recovered to show normal growth patterns through the rest of the study. Reversible effects in the first week are not considered to equate to an "adverse" response. It was concluded that the No-Observed-Adverse-Effect-Level (NOAEL) for COAG-122 was in excess of 1000 mg/kg/day (the limit dose) for reproductive performance and offspring growth and survival in the CD rat following oral gavage administration in a standard screening test.

Results: F1 generation

Effect levels (F1)

Dose descriptor:

NOAEL

Generation:

F1

Effect level:

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

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: Reproductive NOAEL

Overall reproductive toxicity

Reproductive effects observed:

not specified

Any other information on results incl. tables

Signs of offspring

The offspring of female 79 appeared scattered and cold to touch from Day 1 of age. These offspring were small at birth, and failed to thrive after birth. The female and her offspring were therefore killed for welfare reasons on Day 3 of lactation.

Otherwise, the general appearance and behaviour of the offspring were not affected by maternal treatment.

Litter size, survival indices and sex ratio

One female which received COAG-122 at 1000 mg/kg/day (4F 79) was killed on Day 3 of lactation for welfare reasons because of poor litter condition/pup deaths and termination of the litter. The assessment of litter responses detailed below is made based on the 10, 10, 10 and 10 litters in each of groups 1, 2, 3 and 4, respectively, which gave birth to a live litter.

The mean number of implantations, the live litter size on Day 1, the offspring survival up to Day 7 of age and sex ratio showed no adverse effect of parental treatment at 100, 300 or 1000 mg/kg/day.

Bodyweight

Offspring bodyweight on Day 1 of age and subsequent bodyweight gain up to Day 7 of age was similar to control values for the litters of females receiving COAG-122 at 100 , 300 or 1000 mg/kg/day.

Offspring macropathology

Macroscopic examination of offspring which died or were killed for welfare reasons generally confirmed the last in-life finding of no milk present in stomach. No other macroscopic abnormalities were found amongst these animals.

Macroscopic examination of offspring killed at scheduled termination on Day 7 of age revealed no abnormality.

Applicant's summary and conclusion

Conclusions:

Adult animals dosed with COAG-122 at doses of up to 1000 mg/kg/day showed minimal evidence of adverse response. During the first week of treatment effects were seen on male bodyweight at the high dose level but the animals quickly recovered to show normal growth patterns through the rest of the study. Reversible effects in the first week are not considered to equate to an "adverse" response.

Conclusion
It was concluded that the No-Observed-Adverse-Effect-Level (NOAEL) for COAG-122 was in excess of 1000 mg/kg/day (the limit dose) for reproductive performance and offspring growth and survival in the CD rat following oral gavage administration in a standard screening test.

Executive summary:

The influence of COAG-122 solution, on reproductive performance and development in the Crl:CD(SD) rat was assessed in an OECD 421 screening study. Three groups of ten male and female rats received COAG-122 (supplied in solution) by gavage at doses of 100, 300 or 1000 mg/kg/day. The adults were treated daily for a minimum of 15 days before pairing until Day 6 after the birth of the F1 generation. A similarly constituted Control group received the vehicle, purified water, at the same volume-dose (10 mL/kg/day) throughout the same period.

The F1 generation received no direct administration of the test substance; any exposure was in utero or via breast milk.

During the study, clinical condition, bodyweight, food consumption, macroscopic and microscopic pathology investigations were undertaken in all adults. Oestrous cycles and gestation length were assessed and parturition observations were performed for F0 females.

Organ weights were recorded for F0 males only. The clinical condition, litter size and survival, sex ratio and bodyweight of all offspring were assessed.