Registration Dossier

Toxicological information

Repeated dose toxicity: oral

Currently viewing:

Administrative data

Endpoint:
repeated dose toxicity: oral, other
Remarks:
Dose Range Finding Study for Reproduction / Developmental Toxicity Screening Test after Oral Administration in Wistar Rats (OECD 421/422)
Type of information:
experimental study
Adequacy of study:
key study
Study period:
31 July 2018 - 07 January 2019
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2018
Report Date:
2019

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
other: DRF study in rats for OECD 421/422
Version / remarks:
The details in the study report results relevant to study number 183812 (Combined Repeated Dose Oral Toxicity Study with the Reproduction / Developmental Toxicity Screening Test in Wistar Rats with 4,6-dichloro-N-(1,1,3,3-tetramethylbutyl)- 1,3,5-triazin-2-amine).
GLP compliance:
no
Remarks:
DRF study; GLP not required
Limit test:
no

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
solid
Details on test material:
Colour: light yellow
Specific details on test material used for the study:
SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: Suqian Unitech Co., LTD; 2018041002
- Purity: ≥ 99.29 %

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: room temperature, protected from light
- Stability under test conditions: stable
- Solubility and stability of the test substance in the solvent/vehicle: stable in the corn oil

TREATMENT OF TEST MATERIAL PRIOR TO TESTING
- Treatment of test material prior to testing: The test item formulations were prepared freshly on each administration day before the administration procedure. The test item were emulsified in corn oil.

Test animals

Species:
rat
Strain:
Wistar
Details on species / strain selection:
healthy Wistar rats, Crl: WI(Han) (Full Barrier)
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Charles River, 97633 Sulzfeld, Germany
- Females (if applicable) nulliparous and non-pregnant: [yes]
- Age at study initiation: males: 14 - 15 weeks old, females: 14 - 15 weeks old.
- Weight at study initiation: males: 384 - 406 g (mean: 393.42 g, ± 20 % = 314.73 – 472.10 g); females: 228 - 256 g (mean: 238.92 g, ± 20 % = 191.13 – 286.70 g)
- Housing: Full barrier in an air-conditioned room; The animals were kept in groups of 3 animals / sex / cage in IVC polysulphoge cages (double decker, type GR1800) on Altromin saw fibre bedding (except during the mating period when one female was paired with one male and during gestation/lactation period when females were housed individually in type III H IVC polysulphone cages)
- Diet: Free access to Altromin 1324 maintenance diet for rats and mice
- Water: Free access to tap water, sulphur acidified to a pH of approximately 2.8 (drinking water, municipal residue control, microbiological controls at regular intervals)
- Acclimation period: at least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 3 °C
- Humidity (%): 55 ± 10 %
- Air changes (per hr): 10 x / hour
- Photoperiod (hrs dark / hrs light): 12 hours light, 12 hours dark

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
corn oil
Details on oral exposure:
PREPARATION OF DOSING SOLUTIONS:
The vehicle was selected based on the test item’s characteristics. The test item formulations were prepared freshly on each administration day before the administration procedure. The test item were emulsified in corn oil. The test item were weighed into a tared plastic vial on a precision balance. The dose formulations were prepared by adding the required volume of corn oil to give the appropriate final concentration of the test item. The formulation was homogenized by subjecting it to a water bath (40°C) until visual homogeneity was achieved (at least 30 min). Formulates were kept under magnetic stirring during the daily administration. The vehicle was used as control item.

VEHICLE
- Justification for use and choice of vehicle (if other than water): The test item does not make a solution or suspension with water, so the corn oil has to be used.
- Amount of vehicle (if gavage): 4 mL/kg body weight
- Lot/batch no. (if required): MKCD 1021
Analytical verification of doses or concentrations:
no
Details on analytical verification of doses or concentrations:
A dose formulation analysis was not performed in this study.
Duration of treatment / exposure:
The animals were treated with the test item formulation or vehicle on 7 days per week for a period of 54 days, i.e. during 14 days of pre-mating and maximum 14 days of mating in both males and females, during the gestation period and up to post-natal day 3 in females. Males were dosed after the mating period until the minimum total dosing period of 28 days was completed.
Frequency of treatment:
7 days per week
Doses / concentrationsopen allclose all
Dose / conc.:
100 mg/kg bw/day (nominal)
Remarks:
LD/low dose
Dose / conc.:
300 mg/kg bw/day (nominal)
Remarks:
MD/medium dose
Dose / conc.:
1 000 mg/kg bw/day (nominal)
Remarks:
HD/high dose, Test item-related mortality occurred in 3/3 males and 3/3 females of the HD group. Subsequently, the animals were not treated with the test item on study day 3 and the high dose was reduced from 1000 mg/kg body weight/day to 600 mg/kg body weight/day from study day 4 onwards.
Dose / conc.:
600 mg/kg bw/day (nominal)
Remarks:
HD(high dose); Test item-related mortality occurred in 3/3 males and 3/3 females of the HD group. Subsequently, the animals were not treated with the test item on study day 3 and the high dose was reduced from 1000 mg/kg body weight/day to 600 mg/kg body weight/day from study day 4 onwards.
Dose / conc.:
500 mg/kg bw/day (nominal)
Remarks:
HID/high intermediate dose. Due to the high mortality rate in the HD group and also after dose reduction from 1000 mg/kg body weight/day to 600 mg/kg body weight/day, an additional group was inserted and treated with 500 mg/kg body weight/day (HID) to investigate whether 500 mg/kg body weight/day is a feasible dose for higher tier studies.
No. of animals per sex per dose:
3 males
3 females
Control animals:
yes, concurrent vehicle
Details on study design:
- Dose selection rationale: In consultation with the sponsor the following doses were selected for the 3 dose groups (LD = low dose 100 mg/kg bw, MD = medium dose 300 mg/kg bw, HD = high dose 1000mg/kg bw) and 1 control group (C). Due to the high mortality rate in the HD group and also after dose reduction from 1000 mg/kg body weight/day to 600 mg/kg body weight/day, an additional group was inserted and treated with 500 mg/kg body weight/day (HID) to investigate whether 500 mg/kg body weight/day is a feasible dose for higher tier studies.
- Rationale for animal assignment (if not random): randomisation was performed with IDBS Workbook 10.1.2 software

Examinations

Observations and examinations performed and frequency:
DETAILED CLINICAL OBSERVATIONS: Yes; The health condition of the animals was recorded. Twice daily all animals were observed for morbidity and mortality except on weekends and public holidays when observations were made once daily. Females showing signs of abortion or premature delivery prior to the scheduled scarification of the animals were sacrificed and subjected to a thorough macroscopic examination.
Clinical observations included spontaneous activity, lethargy, recumbent position, convulsions, tremors, apnoea, asphyxia, vocalisation, diarrhoea, changes in the skin and fur, eyes and mucous membranes (salivation, discharge), piloerection and pupil size. Changes in gait, posture, response to handling as well as the presence of clonic or tonic movements, stereotypes, difficult or prolonged parturition or bizarre behaviour were recorded.
- Time schedule: at least once a day

BODY WEIGHT: Yes
- Time schedule for examinations: The body weight was recorded once before the assignment to the experimental groups, on the first day of administration and weekly during the treatment period as well as at the end of the study. During pregnancy, females were weighed on gestation days (GD) 0, 7, 14 and 20 and within 24 hours of parturition (day 0 post-partum) as well as day 4 post-partum along with pups. Animals prematurely sacrificed were weighed prior to the sacrifice.

FOOD CONSUMPTION AND COMPOUND INTAKE:
- Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: Yes; Food consumption was measured weekly on the corresponding days of the body weight measurements after the beginning of the dose administration. Food consumption was not measured during the mating period in males and females and the post-mating period in males.


Sacrifice and pathology:
GROSS PATHOLOGY: Yes (see table below)
The males were sacrificed after completion of the mating period on day 29 with the exception of animal nos. 10, 11, and 12 of the high dose group which were sacrificed during the premating phase due to animal welfare reasons (no.10 on PMD 7, no. 11 on PMD 3 and no. 12 on PMD 8). The females were sacrificed along with their pups on post-natal day 4 with the exception of animal nos. 22, 23, and 24 of the high dose group which were sacrificed due to animal welfare reasons during the premating phase (no. 22 and no. 23 on PMD 8 and no. 24 on PMD 7). Non-pregnant females were sacrificed on day 26 (female no. 17 of the LD group).

Dead pups and pups sacrificed on day 4 post-partum or shortly thereafter were carefully examined externally for gross abnormalities.

Non-pregnant females were sacrificed on study day 26 using the sperm-positive vaginal smear as an evidence of mating.

All animals were subjected to a detailed gross necropsy which includes careful examination of the external surface of the body, all orifices and the cranial, thoracic and abdominal cavities and their contents.
Special attention was paid to the organs of the reproductive system. The ovaries, uterus with cervix, vagina, testes, epididymides, accessory sex organs (prostate, seminal vesicles with coagulating glands as a whole), and all organs showing macroscopic lesions of all adult animals were preserved in 4% neutral-buffered formaldehyde, except for testes and epididymides which were preserved in modified Davidson’s Solution and then transferred in 70 % ethanol.

All macroscopic findings were recorded and organs showing gross abnormalities and all organs listed were preserved in 4 % neutral-buffered formaldehyde except testes and epididymides which were fixed in Modified Davidson’s fixative for approximately 24 hours before they were transferred to 70 % ethanol.

All animals found dead and/or intercurrently sacrificed were subjected to a gross necropsy.

The number of implantation sites and corpora lutea was recorded for each parental female at necropsy. If appropriate and possible, the number of corpora lutea and implantation sites was recorded for any females sacrificed 26 days after the end of the mating period with no evidence of mating and or on day 26 post-coitum due to non-delivery.

Organ Weights
Reproductive organs from all animals were weighed (testes, epididymides, prostateseminal vesicle with coagulating glands as whole, ovaries, uterus with cervix).
Organ weights from animals found dead or euthanized due to animal welfare were not taken.
Other examinations:
Litter Observations

The duration of the gestation was recorded and was calculated from day 0 of the pregnancy. Each litter was examined as soon as possible after delivery of the dam to establish the number and sex of pups, stillbirths, live births, runts and the presence of gross abnormalities.

Live pups were counted and sexed and litters weighed within 24 hours of parturition (day 0 post-partum) and on day 4 post-partum. Live pups were identified by writing numbers on the back with the help of a permanent marker or by tattooing. In addition to the observations of parent animals, any abnormal behaviour of the offspring was recorded.
Statistics:
Mean and standard deviations

Results and discussion

Results of examinations

Clinical signs:
effects observed, treatment-related
Description (incidence and severity):
For a detailed description of the findings see Table 6, Table 7, Table 8, Table 9, Table 29 and Table 30.

Male no. 10 which was euthanized in a moribund condition on premating day showed the clinical signs of a stilted gait, diarrhea, sunken flanks, dehydration, piloerection, moderately reduced spontaneous activity, and nasal discharge between PMD 3 and PMD 7 (day of sacrifice).

Prematurely sacrificed HD male no. 11 was observed with signs of piloerection, diarrhea, sunken flanks, dehydration and a stilted gait on the day of sacrifice (PMD 3).

Male no. 12 of the HD group which was sacrificed due to animal welfare reasons on PMD 8 was observed with clinical signs of piloerection, diarrhea, sunken flanks, dehydration, stilted gait, chromodacryorrhea, and a bloody wound on the tail root between PMD 3 and PMD 8 (day of sacrifice).
Prematurely sacrificed HD female no. 22 showed the clinical signs of diarrhea, dehydration, sunken flanks, stilted gait, reduced spontaneous activity, hypothermia, and a bloody wound on the tail root between PMD 3 and the day of sacrifice (PMD 8).

Female no. 23 of the HD group was euthanized in a moribund condition with clinical signs of diarrhea, sunken flanks, stilted gait, dehydration, and hypothermia between PMD 3 and PMD 8 (day of sacrifice).

Prematurely sacrificed HD female no. 24 was observed with clinical signs of stilted gait, sunken flanks, diarrhea, dehydration, reduced spontaneous activity between PMD 3 and the day of sacrifice (PMD 7).

3/3 females of the MD group were observed with the clinical sign of sunken flanks on PMD 3. As this finding occurred on a single day it is not considered toxicologically relevant.

The clinical sign of moving the bedding was observed transiently in 2/3 males of the LD group, 3/3 males of the MD group, 2/3 males of the HD group and 1/3 females of the control group, 1/3 females of the LD group, 3/3 females of the MD group, and 3/3 females of the HD group. The clinical sign of increased salivation was seen in 1/3 males of the LD group, 3/3 males of the MD group, 2/3 males of the HD group, and1/3 females of the LD group, 3/3 females of the MD group, and 3/3 females of the HD group. As both signs were observed in short timely relation to test item administration or in anticipation thereof they were considered to be a sign of discomfort or a local reaction of the test item.

Low incidences of clinical signs like hairless areas, crusts, or wounds in few animals of the control or test item-treated groups were seen without dose dependency and were considered as incidental.

None of the females showed clinical signs of abortion or premature delivery.

The clinical signs in males of the HID group which were sacrificed prematurely included diarrhea, piloerection, reduced spontaneous activity, hypothermia, stilted gait, sunken flanks, hunched posture, dehydration, nasal discharge, and abnormal breathing.
Females were observed with clinical signs of piloerection, diarrhea, hunched posture, and reduced spontaneous activity in the second half of the treatment period.

The clinical sign of moving the bedding was observed transiently in 3/3 males and 3/3 females of the HID group. The clinical sign of increased salivation was seen in 2/3 females of the HID group. Both signs were observed in relation to test item administration or in anticipation thereof they were considered to be a sign of discomfort or a local reaction of the test item.
Mortality:
mortality observed, treatment-related
Description (incidence):
For a summary see Table 4 and Table 5.

During the treatment period of this study mortality occurred in 3/3 males of the HD group, 3/3 females of the HD group, and 3/3 males of the HID group.

Male no. 11 of the HD group was euthanized in a moribund condition on premating day 3. HD male no. 10 was sacrificed due to animal welfare reasons on premating day 7, and male no. 12 of the HD group was euthanized due to animal welfare reasons on premating day 8.

HD female no. 24 was sacrificed in a moribund condition on premating day 7 and female nos. 22 and 23 of the HD group were euthanized due to animal welfare reasons on premating day 8.

Male no. 25 was euthanized in a moribund condition on treatment day 9, male no. 26 was euthanized due to animal welfare reasons on treatment day 10, and male no. 27 was euthanized in a moribund condition on treatment day 28.

All remaining animals (control, LD, and MD) survived their scheduled study period.
Body weight and weight changes:
effects observed, treatment-related
Description (incidence and severity):
For a detailed description of the findings see Table 10 to Table 16 and Table 31 to Table 38.

Treatment with the test item had no toxicologically relevant effect on body weight development of male and female animals of the LD group during the whole treatment period of this study.

Mean body weight of males of the HD group was lower compared to the controls on premating day 7 (deviation of the HD group from the control group: 17%) and compared to the body weight on premating day 1. HD animals showed mean body weight loss of 57 g during the first week of treatment.

Mean body weights of the MD males were marginally lower when compared to the control group from premating day 7 until the end of the treatment period (mean body weight of the MD group was 3 % - 6 % lower compared to the mean body weight of the controls). Accordingly, males of the MD group showed a lower mean body weight gain compared to the controls throughout the treatment period (body weight gain of MD males was 34 % lower compared to the control group considering premating day 1 to postmating day 14).

Females of the HD group showed a lower mean body weight on premating day 7 when compared to the female control group (deviation from control: 13%) and premating day 1. HD females showed a mean body weight loss of 35% during the first week of treatment.

Mean values of body weigh were slightly lower in the females of the MD group when compared to the controls from premating day 14 onwards (mean body weight of MD females was 4 % - 12 % below mean body weight of the control group. Mean body weight gain was comparable between MD females and females of the control group with the exception of the second week of treatment (premating day 7 - premating day 14) where females of the MD group showed a mean body weight loss of 15 g compared to a mean body weight gain of 5 g in the control group.

Males of the HID group were observed with a lower mean body weight on day 7 compared to day 1 of body weight measurement (mean bw day 1: 427 g, mean bw day 7: 386 g). Females of the HID group showed a body weight loss during the first week of treatment (mean bw day 1: 223 g, mean bw day 7: 218).
Food consumption and compound intake (if feeding study):
effects observed, treatment-related
Description (incidence and severity):
For a detailed description of the findings see Table 18, Table 19, Table 39 and Table 40.

Mean food consumption of was not affected in males of the LD and MD group when compared to the control males throughout the premating period. Food consumption of HD males was lower compared to the control group during the first week of treatment (deviation of the HD group from controls: 84%). However, this value has to be considered carefully as food consumption was available from only 2 males from treatment day 3 onwards.

Mean food consumption of females was not affected by treatment with the test in the LD and MD group compared to the female control group throughout the treatment period (premating, gestation, and lactation). Females of the HD group showed lower mean food consumption compared to the controls during the first week of treatment (deviation of the HD group from the control group: 36%).
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
Immunological findings:
not examined
Organ weight findings including organ / body weight ratios:
effects observed, non-treatment-related
Description (incidence and severity):
For a detailed description of the findings see Table 27 and Table 48.

There were no test item-caused differences in the weight of reproductive organs between treated groups and the respective control group.
However, mean prostate weight of LD males was slightly lower compared to controls (deviation from control: 20%). Due to the lack of dose dependency this trend was not considered test item-related.

Mean ovary weight was shown to be slightly higher in the females of the LD (deviation from control: 18%) and MD (deviation from control: 21%) group compared to the mean ovary weight of the control females. Mean uterus weight was slightly higher in LD females compared to the control group (deviation from control: 13%). However, these effects are not considered toxicologically relevant as female reproductive organs may undergo changes depending on the stage of the estrous cycle.
Gross pathological findings:
effects observed, treatment-related
Description (incidence and severity):
For a detailed description of the findings see Table 26.

Macroscopic findings that could be attributed to treatment with the test item consisted of an abnormal colored stomach (white and spotted) of male no. 11 which was euthanized due to animal welfare reasons and a stomach containing few (2-6) masses of male no. 12 which was euthanized in a moribund condition.

Single findings of an enlarged spleen in 1/3 females of the control group, a thymus with several (7-12) red foci in 1/3 males of the MD group, a small thymus in 1/3 females of the MD group, kidneys with an abnormal surface (crater) in 1/3 females of the MD group were within the range of normal background alterations and thus are not considered toxicologically relevant.

Male no. 26 of the HID group which was euthanized in a moribund condition on treatment day 10 was observed with a fluid filled (clear) and dark/white spotted stomach.
Neuropathological findings:
not examined
Histopathological findings: non-neoplastic:
not examined
Histopathological findings: neoplastic:
not examined
Other effects:
effects observed, non-treatment-related
Description (incidence and severity):
Litter Data
For a detailed description of the findings see Table 20 and Table 41.
Treatment with the test item had no toxicologically relevant effects on pup associated parameters including total number of pups, still births, runts, and sex ratio.
Litters were delivered by all dams of the control and the MD group and by 2/3 females of the LD group. Treatment with the test item had no effect on the total number of pups when comparing treated groups with the control group on PND 0 and PND 4.
There were no considerable differences in the sex ratio between test item-treated groups and the control group. No runts and still births were observed in any of the test item-treated groups or the control group.

Litter Weight Data
For a detailed description of the findings see Table 21 and Table 42.
Treatment with the test item had no toxicologically relevant effect on litter weight data.
Mean pup weight was comparable between pups of the test item-treated groups (LD and MD) and the controls.
In accordance with a slightly lower number of live male pups in the MD group on PND 0 and PND 4 male litter weight and total litter weight was slightly lower in the MD group when compared to the controls. However, this was not considered toxicologically relevant.

Precoital Interval and Duration of Gestation
For a detailed description of the findings see Table 22 and Table 43.
Treatment with the test item had no toxicologically relevant effect on precoital interval and the duration of gestation.
Mean precoital interval was slightly lower in the MD group when compared to the controls. However, this variation is within the normal range of variation. Mean duration of gestation was comparable between test item-treated groups and the control group.

Pre- and Post-Natal Data
For a detailed description of the findings see Table 23 and Table 44.
Treatment with the test item had no toxicologically relevant effect of pre- and postnatal data in this study. Mean values of corpora lutea, implantation sites, alive pups on PND 0 and PND 4, pre-implantation loss, and post-implantation loss. Differences were within the normal range of variation of this strain.

Reproductive Indices
For a detailed description of the findings see Table 24 and Table 45.
There were no treatment-related effects on the reproductive indices including copulation index, fertility index, delivery index and viability index when comparing the LD and the MD group with the control group.
Slightly lower fertility index of the LD group (66.67 %) compared to the control group was based on one non-pregnant female of the LD group. This was considered incidental and thus not toxicologically relevant.
Apart from the fertility index of 66.67 % in the LD group all indices were 100 % in the control, the LD, and MD group.

Pup Survival Data
For a detailed description of the findings see Table 25 and Table 46.
Survival of live born pups was not different between the control group and groups treated with the test item. All live born pups survived from PND 1 to PND 4 in all groups.

Pup External Findings
For a detailed description of the findings see Table 47.
One pup of the control group (dam no. 14, pup no. 3) was observed to be pale on PND 1. As this finding occurred in the control group it is not considered toxicologically relevant. There were no further external macroscopic findings in any of the pups of the control or the test item-treated groups.

Effect levels

Dose descriptor:
other: DRF study
Effect level:
0 mg/kg bw/day (nominal)
Sex:
male/female
Basis for effect level:
other: DRF study

Target system / organ toxicity

Critical effects observed:
no

Applicant's summary and conclusion

Conclusions:
Based on the data generated from this dose range finding study, dose levels of 40, 120 and 360 mg/kg bw per day are suggested for the subsequent main Combined Repeated Dose Toxicity Study with Reproductive/Developmental Toxicity Screening Test (OECD 422) with 4,6-dichloro-N-(1,1,3,3-tetramethylbutyl)-1,3,5-triazin-2-amine.
Executive summary:

In a dose range finding study for a reproduction/developmental toxicity screening test (183813), 4,6-dichloro-N-(1,1,3,3-tetramethylbutyl)-1,3,5-triazin-2-amine (99.29%) was administered to 5 groups of Wistar rats (3 animals/sex/group) by gavage in corn oil at dose levels of 0, 100 (LD), 300 (MD), and 1000 (HD) or 600 (HD) mg/kg body weight/day and additionally 500 (HID) mg/kg body weight/day, 7 days per week. The animals were treated for a period of 54 days, i.e. during 14 days of pre-mating and maximum 14 days of mating in both males and females, during the gestation period and up to post-natal day 3 in females. Males were dosed after the mating period until the minimum total dosing period of 28 days was completed.

Test item-related mortality occurred in 3/3 males and 3/3 females of the HD group. Subsequently, the animals were not treated with the test item on study day 3 and the high dose was reduced from 1000 mg/kg body weight/day to 600 mg/kg body weight/day from study day 4 onwards. Males of the additional group dosed with 500 mg/kg body weight/day were all euthanized in a moribund condition between treatment day 8 and 10.

Severe clinical findings in the HD group caused by treatment with 4,6-dichloro-N-(1,1,3,3-tetramethylbutyl)-1,3,5-triazin-2-amine were reduced spontaneous activity, diarrhea, dehydration, stilted gait and sunken flanks.  Gross pathological changes which may be caused by treatment with the test item were an abnormal colored stomach in one animal of the HD group and masses in the stomach of another HD animal.  No mortality or clinical symptoms indicating systemic toxicity were observed at dose levels of 100 and 300 mg/kg body weight/day.  At a dose level of 300 mg/kg body weight/day (MD) a slightly attenuated body weight gain was observed in males and females.

Under the conditions of the study performed, it is assumed that 4,6-dichloro-N-(1,1,3,3-tetramethylbutyl)-1,3,5-triazin-2-amine has no toxic effects on male and female reproductive performance such as gonadal function, mating behavior, conception, development of the conceptus and parturition as well as pup-related parameters.

Based on the data generated from this dose range finding study, dose levels of 40, 120 and 360 mg/kg bw per day are suggested for the subsequent main Combined Repeated Dose Toxicity Study with Reproductive/Developmental Toxicity Screening Test (OECD 422; 183812) with 4,6-dichloro-N-(1,1,3,3-tetramethylbutyl)-1,3,5-triazin-2-amine.