1,5-Pentanediamine, 2-methyl-, reaction products with 2-ethyl-1,4-butanediamine and glycidyl tolyl ether

Developmental toxicity / teratogenicity

Administrative data

Endpoint:

developmental toxicity

Type of information:

experimental study

Adequacy of study:

key study

Study period:

05 May 2017 - 03 July 2017

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

comparable to guideline study with acceptable restrictions

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2019

Report date:

2019

Materials and methods

Test guideline

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 414 (Prenatal Developmental Toxicity Study)

Version / remarks:

designed as dose range finding study

Deviations:

yes

Remarks:

designed as dose range finding study, thus, only limited ex aminations

GLP compliance:

yes (incl. QA statement)

Limit test:

no

Test material

Test material information

Constituent 1

Reference substance name:

1-[(5-amino-4-methylpentyl)amino]-3-(2-methylphenoxy)-propan-2-ol

Cas Number:

1617528-43-4

Molecular formula:

C16H28N2O2

IUPAC Name:

1-[(5-amino-4-methylpentyl)amino]-3-(2-methylphenoxy)-propan-2-ol

Constituent 2

Reference substance name:

1,1'-[(2-Methyl-1,5-pentanediyl)diimino]bis[3-(2-methylphenoxy)-propan-2-ol]

Cas Number:

1617528-45-6

Molecular formula:

C26H40N2O4

IUPAC Name:

1,1'-[(2-Methyl-1,5-pentanediyl)diimino]bis[3-(2-methylphenoxy)-propan-2-ol]

impurity 1

Reference substance name:

Mephenesin

EC Number:

200-427-4

EC Name:

Mephenesin

Cas Number:

59-47-2

Molecular formula:

C10H14O3

IUPAC Name:

3-(2-methylphenoxy)propane-1,2-diol

impurity 2

Reference substance name:

1,3-bis(2-methylphenoxy)propan-2-ol

Cas Number:

17181-49-6

Molecular formula:

C17H20O3

IUPAC Name:

1,3-bis(2-methylphenoxy)propan-2-ol

impurity 3

Reference substance name:

2-methylpentane-1,5-diamine

EC Number:

239-556-6

EC Name:

2-methylpentane-1,5-diamine

Cas Number:

15520-10-2

Molecular formula:

C6H16N2

IUPAC Name:

2-methylpentane-1,5-diamine

impurity 4

Reference substance name:

3,3’-(5-(2-hydroxy-3-(o-tolyloxy)propylamino)-4-methylpentylazanediyl)bis(1-(o-tolyloxy)propan-2-ol)

Molecular formula:

C36H52N2O6

IUPAC Name:

3,3’-(5-(2-hydroxy-3-(o-tolyloxy)propylamino)-4-methylpentylazanediyl)bis(1-(o-tolyloxy)propan-2-ol)

Test material form:

liquid

Test animals

Species:

rat

Strain:

Wistar

Remarks:

Wistar Han'": Rccl-lant'': WIST

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Envigo RMS (UK) Ltd., Oxon, UK
- Age at study initiation: time-mated females, delivered prior to day 3 of gestation
- Housing: individually in solid floor polypropylene cage with stainless steel mesh lids and softwood flake bedding
- Diet (e.g. ad libitum): Rodent 2018C Teklad Global Certified Pelleted Diet, ad libitum
- Water (e.g. ad libitum): tap water, ad libitum
- Acclimation period: day of delivery until day 3 of gestation

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 3
- Humidity (%): 30 - 70
- Air changes (per hr): at least 15
- Photoperiod (hrs dark / hrs light): 12/12

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

polyethylene glycol

Remarks:

PEG400

Analytical verification of doses or concentrations:

no

Details on mating procedure:

- Impregnation procedure:purchased timed pregnant
- Proof of pregnancy: vaginal plug / sperm in vaginal smear referred to as day 0 of pregnancy

Duration of treatment / exposure:

17 d (day 3 to 19 of gestation)

Frequency of treatment:

daily from day 3 to 19 of gestation

Duration of test:

20 d

Doses / concentrations

Doses / concentrations 1

Dose / conc.:

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

Doses / concentrations 2

Dose / conc.:

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

Doses / concentrations 3

Dose / conc.:

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

Doses / concentrations 4

Dose / conc.:

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

No. of animals per sex per dose:

5

Control animals:

yes, concurrent vehicle

Examinations

Maternal examinations:

- Time schedule:
mortality: Twice daily, early and late during the working period.
clinical signs: Individual clinical observations will be performed immediately before dosing, up to 30 minutes after dosing and one hour after dosing. In addition, post dosing observations will also be performed at approximately 4 hours after dosing during the normal working day (excluding weekends and Public Holidays). All observations will be recorded.

BODY WEIGHT: Yes
- Time schedule for examinations: Individual body weights will be recorded on Days 3, 4, 5, 8, 11, 14, 17 and 20 of gestation.

FOOD CONSUMPTION:
Dietary intake will be recorded for individual animals on Days 3, 5, 8, 11, 14, 17 and 20 of gestation.

FOOD EFFICIENCY:
- Body weight gain in kg/food consumption in kg per unit time X 100 calculated as time-weighted averages from the consumption and body weight gain data: No

WATER CONSUMPTION: Yes
- Time schedule for examinations: Water consumption will be monitored daily by visual inspection of water bottles.

POST-MORTEM EXAMINATIONS: Yes
- Sacrifice on gestation day #20
- gross necropsy

Ovaries and uterine content:

The uterus of each adult female will be examined and where possible the following recorded;
i. Pregnancy status
ii. Number of corpora lutea
iii. Gravid uterus weight
iv. Number, status* and intra-uterine position of implantations
* = live fetus, dead fetus, late intra-uterine death or early intra-uterine death

Fetal examinations:

For each fetus/placenta, where possible, the following will be recorded:
i. External fetal abnonnalities
11. Fetal weight
iii. Fetal sex
iv. Placental weight

Results and discussion

Results: maternal animals

General toxicity (maternal animals)

Clinical signs:

effects observed, treatment-related

Description (incidence and severity):

Between Day 5 to 7 of gestation, the majority of females treated with 75 or 150 mg/kg bw/day showed instances of increased salivation and/or noisy respiration.
Incidental findings included one 75 mg/kg bw/day females showing observations of deceased respiratory rate, pilo-erection and diarrhoea between Days 5 to 7 with noisy respiration between Days 5 to 10 and 12 to16.

Mortality:

mortality observed, treatment-related

Description (incidence):

One 150 mg/kg bw/day female was found dead shortly after dosing on Day 5 of Gestation, there were no adverse effects on body weight, food consumption or clinical signs prior to death. Necropsy findings revealed darkened liver and kidneys, with dark patches in the lungs.
There were no further unscheduled deaths on the study.

Body weight and weight changes:

effects observed, treatment-related

Description (incidence and severity):

All female treatment groups exhibited lower body weight gains following the first dose (Day 3), with improvement evident thereafter.
During the first Week of treatment lower overall accumulative body weight gains were evident in females treated with 75 or 150 mg/kg bw/day in relation to controls. However, during the latter stage of gestation, overall body weight gains return to levels similar to controls. Body weight gains when adjusted for gravid uterus weight, revealed lower body weight gain of 17% and 40%, in females treated with 75 and 150 mg/kg bw/day, respectively. No such effect was detected at 25 mg/kg bw/day.

Food consumption and compound intake (if feeding study):

effects observed, treatment-related

Description (incidence and severity):

Instances of marginally lower food consumption were noted in 75 and 150 mg/kg bw/day dose groups between Days 3 to 8 and 3 to 5 of gestation, respectively. Thereafter food consumptions were generally similar to controls.

Food efficiency:

not examined

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

no effects observed

Description (incidence and severity):

Visual inspection of water consumptions revealed no adverse effects.

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:

not examined

Gross pathological findings:

no effects observed

Description (incidence and severity):

The macroscopic findings in the decedent female (No. 58) findings revealed darkened liver and kidneys, with dark patches in the lungs.
No macroscopic abnormalities were detected in animals examined at study termination.

Neuropathological findings:

not examined

Histopathological findings: non-neoplastic:

not examined

Histopathological findings: neoplastic:

not examined

Maternal developmental toxicity

Number of abortions:

no effects observed

Pre- and post-implantation loss:

effects observed, treatment-related

Total litter losses by resorption:

no effects observed

Early or late resorptions:

no effects observed

Dead fetuses:

no effects observed

Changes in pregnancy duration:

not examined

Description (incidence and severity):

Migrated Data from removed field(s)
Field "Effects on pregnancy duration" (Path: ENDPOINT_STUDY_RECORD.DevelopmentalToxicityTeratogenicity.ResultsAndDiscussion.ResultsMaternalAnimals.MaternalDevelopmentalToxicity.EffectsOnPregnancyDuration): not examined

Changes in number of pregnant:

not examined

Details on maternal toxic effects:

Litter Data
There was no detrimental effect of maternal treatment on litter data as assessed by corpora lutea, numbers of implantations, in-utero offspring survival (as assessed by the mean numbers of early or late resorptions), live litter size and sex ratio at 25, 75 or 150 mg/kg bw/day.
Females treated with 150 mg/kg bw/day showed slightly higher pre-implantation losses when compared to controls. Females at 25 or 150 mg/kg bw/day also showed slightly higher post-implantation losses in comparison with controls, without any dose response.

Litter Placental and Foetal weights
There were no effects of maternal treatment on mean foetal weights, placental weights or litter weight at 25, 75 or 150 mg/kg bw/day.

Effect levels (maternal animals)

Dose descriptor:

other: upper dose level for definitive OECD TG 422 study

Effect level:

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

Based on:

act. ingr.

Basis for effect level:

body weight and weight gain

clinical signs

mortality

Maternal abnormalities

Abnormalities:

not specified

Results (fetuses)

Fetal body weight changes:

no effects observed

Description (incidence and severity):

Migrated Data from removed field(s)
Field "Fetal/pup body weight changes" (Path: ENDPOINT_STUDY_RECORD.DevelopmentalToxicityTeratogenicity.ResultsAndDiscussion.ResultsFetuses.FetalPupBodyWeightChanges): no effects observed

Reduction in number of live offspring:

no effects observed

Changes in sex ratio:

no effects observed

Changes in litter size and weights:

no effects observed

Changes in postnatal survival:

not examined

External malformations:

no effects observed

Skeletal malformations:

not examined

Visceral malformations:

not examined

Details on embryotoxic / teratogenic effects:

External Examinations
There were no findings apparent for foetuses from treated females at external examination on Day 20 of gestation.
At 150 mg/kg bw/day incidental findings included one litter with five pale fetuses and a further one litter contained one fetus with encephaolcoele.

Effect levels (fetuses)

Dose descriptor:

other: upper dose level for definitive OECD TG 422 study

Effect level:

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

Based on:

act. ingr.

Sex:

male/female

Basis for effect level:

other: overall effects

Fetal abnormalities

Abnormalities:

no effects observed

Description (incidence and severity):

only external examinations performed in this dose range finding study

Overall developmental toxicity

Developmental effects observed:

no

Any other information on results incl. tables

Bodyweights

Group		Days
		1	3	4	5	8	11		14		17		20
1	Mean	215.0	229.0	233.8	235.0	240.8	250.0		260.5		282.5		314.3
	SD	15.4	17.2	18.5	15.7	17.5	18.6		21.0		19.8		16.5
	n	4	4	4	4	4	4		4		4		4
2	Mean	211.0	226.6	227.2	229.8	237.8	249.6		261.0		287.4		323.2
	SD	15.6	16.6	19.1	18.6	18.6	19.4		22.1		24.0		29.4
	n	5	5	5	5	5	5		5		5		5
3	Mean	210.0	225.2	226.6	229.8	233.2	241.0		256.2		278.0		311.6
	SD	15.2	16.2	16.3	14.5	15.2	14.4		16.3		18.3		22.6
	n	5	5	5	5	5	5		5		5		5
4	Mean	209.8	223.0	222.3	225.8	229.0	243.3		256.3		280.3		308.0
	SD	17.8	18.8	22.1	19.0	17.6	19.5		19.6		23.3		30.5
	n	4	4	4	4	4	4		4		4		4

 

Body weight gains

Group		Days
		1	3	4	5	8		11		14		17
		3	4	5	8	11		14		17		20
1	Mean	14.0	4.8	1.3	5.8	9.3		10.5		22.0		31.8
	SD	2.2	1.3	3.0	2.2	2.1		3.1		5.0		3.8
	n	4	4	4	4	4		4		4		4
2	Mean	15.6	0.6	2.6	8.0	11.8		11.4		26.4		35.8
	SD	6.8	4.0	2.4	1.4	0.8		3.6		2.5		5.8
	n	5	5	5	5	5		5		5		5
3	Mean	15.2	1.4	3.2	3.4	7.8		15.2		21.8		33.6
	SD	2.2	1.8	2.4	22.3	21.1		3.1		4.4		4.6
	n	5	5	5	5	5		5		5		5
4	Mean	13.3	-0.8	3.5	3.3	14.3		13.0		24.0		27.8
	SD	4.5	3.9	3.5	2.8	2.8		2.2		3.7		8.1
	n	4	4	4	4	4		4		4		4

 

Food consumption (g/rat/day)

Group		Days
		3	5	8	11	14	17
		5	8	11	14	17	20
1	Mean	16.0	14.8	18.3	21.5	19.8	22.1
	SD	1.1	3.9	1.3	2.6	2.0	1.0
	n	4	4	4	4	4	4
2	Mean	18.2	16.7	19.0	22.3	21.4	23.1
	SD	1.9	1.6	1.1	1.6	2.3	0.5
	n	5	5	5	5	5	5
3	Mean	15.1	11.6	16.9	21.3	19.7	22.5
	SD	1.7	4.7	1.3	2.4	2.1	2.6
	n	5	5	5	5	5	5
4	Mean	12.8	14.3	17.7	21.3	19.0	19.1
	SD	1.9	1.8	1.7	2.3	2.5	3.4
	n	4	4	4	4	4	4

 

 

Group		Copora Lutea	Number of Implants	Embryonic/Fetal Deaths			Implantation losses		Live young			Sex Ratio (% male)	Mean Fetal Weight			Mean Placental Weight	Litter Weight	Total Placental Weight
				Early	Late	Total	Pre-	Post-	Male	Female	Total		Male	Female	Combined
1	Mean	13.0	10.5	0.0	0.0	0.0	19.3	0.0	5.0	5.5	10.5	49.0	3.967	3.669	3.817	0.489	40.023	5.118
	SD	1.4	1.3	0.0	0.0	0.0	4.3	0.0	2.6	3.3	1.3	25.8	0.250	0.295	0.255	0.023	4.887	0.538
	n	4	4	4	4	4	4	4	4	4	4	4	4	4	4	4	4	4
2	Mean	14.5	11.8	0.2	0.0	0.2	19.0	1.3	5.2	6.6	11.8	43.2	4.017	3.856	3.933	0.500	46.534	5.908
	SD	1.7	2.2	0.4	0.0	0.4	10.7	3.0	1.9	0.9	1.6	10.9	0.244	0.236	0.233	0.018	7.775	0.944
	n	4	4	5	5	5	4	5	5	5	5	5	5	5	5	5	5	5
3	Mean	13.0	10.2	0.0	0.0	0.0	20.8	0.0	6.0	4.2	10.2	58.1	4.381	4.165	4.297	0.538	43.648	5.448
	SD	1.9	1.6	0.0	0.0	0.0	13.7	0.0	1.6	0.4	1.6	6.9	0.196	0.248	0.194	0.078	5.933	0.937
	n	5	5	5	5	5	5	5	5	5	5	5	5	5	5	5	5	5
4	Mean	14.5	11.0	0.3	0.0	0.3	24.7	2.5	5.3	5.5	10.8	45.2	4.462	4.176	4.281	0.565	45.493	6.153
	SD	1.3	2.9	0.5	0.0	0.5	15.0	5.0	3.6	1.0	3.1	19.5	0.402	0.474	0.447	0.123	10.959	2.402
	n	4	4	4	4	4	4	4	4	4	4	4	4	4	4	4	4	4

 

Applicant's summary and conclusion

Conclusions:

In consideration of the marked body weight effects on the males during the fourteen day repeated dose phase with non-pregnant animals (Phase I) at 200 mg/kg bw/day and the observed effects on body weight gains on females at 150 mg/kg bw/day during the repeated dose phase on pregnant animals (Phase II), these effects were considered not to be so significant to exclude 150 mg/kg bw/day as the high dose level for further investigating. Therefore, dose levels of 0, 25, 75 and 150 mg/kg bw/day are recommended for the forthcoming Oral (Gavage) Combined Repeat Dose Toxicity Study with Reproduction/Developmental Toxicity Screening Test in the Rat (OECD 422) (Study No. HD72JH).

Executive summary:

The purpose of this study was to give a preliminary indication of the effects of repeated oral administration of the test item to rats over a period of fourteen consecutive days (Phase I).

This study was also intended to establish the effects of the test item on the embryonic and fetal development of the rat when administered orally during gestation, including organogenesis (Phase II). The results of the second phase may identify potential effects of the test item on the development of the embryo/fetus in utero. The study was designed to provide toxicological data sufficient to assist in dose level selection for subsequent longer term toxicological evaluation of the test item in the rat and may also provide suitable information to assist in dose level selection for any subsequent oral pre-natal investigation of developmental toxicity (Study No. HD72JH). Clinical observations, body weight changes, food and water consumption and gross pathology will be monitored, for any adverse effects resulting from

exposure to the test item.

 

In Study Phase I, the test item was administered by oral gavage to three groups, each of five male and five female Wistar Han: RccHan: WIST strain rats, for fourteen consecutive days, at dose levels of 25, 100 and 200 mg/kg bw/day. A control group of five males and five females was dosed with vehicle alone (Polyethylene glycol 400) over the same treatment period.

Clinical signs, body weight development, food consumption and water intake were monitored during the study and all animals were subjected to a gross necropsy examination at termination.

In Study Phase II, the test item was administered once daily by oral gavage to three groups, each of five time-mated Wistar Han: RccHan: WIST strain female rats from Day 3 to Day 19 of gestation at dose levels of 25, 75 and 150 mg/kg bw/day. A control group of five timemated females received vehicle alone (Polyethylene glycol 400) over the same treatment period. All females were killed on Day 20 of gestation.

Clinical signs, body weight development and food consumption were monitored during the study and all animals were subjected to gross necropsy examination including examination of the uterine contents and gross external necropsy of fetuses.

 

Results

Phase I

Mortality

There were no unscheduled deaths during the study phase.

 

Clinical Observations

At 200 or 100 mg/kg bw/day animals of either sex exhibited increased post dosing salivation between Days 7 and 13 of treatment.

 

Body Weight

There were adverse effects on body weight development for animals receiving 200 mg/kg bw/day. No such effects were detected in animals of either sex at 25 or 100 mg/kg bw/day.

 

Food Consumption

There were reduced food consumptions and food conversion efficiency for animals of either sex at 200 mg/kg bw/day.

 

Water Consumption

There were no adverse treatment-related effects in water intake compared to controls.

 

Necropsy

Treatment-related macroscopic findings were confined to enlarged and darkened spleens observed in four 200 mg/kg bw/day females. There were no macroscopic findings in the remaining animals.

 

Phase II

Mortality

One female dosed with 150 mg/kg bw/day was found dead on Day 5 of gestation, showing no adverse effects prior to the death. Due to this isolated incidence at this dose level the death was considered to be of no toxicological significance.

There were no further unscheduled deaths during Phase II of the study.

 

Clinical Observations

The majority of females treated with 75 or 150 mg/kg bw/day showed instances of increased salivation and/or noisy respiration.

 

Body Weight

At 150 or 75 mg/kg bw/day body weight gains and overall accumulated body weight gains showed instances of lower weight gains compared to controls, however, improvement was evident by the end of the treatment period. Body weight gains when adjusted for gravid uterus weight revealed lower group mean body weight gains compared to control at these dose levels.

No such effects were detected at 25 mg/kg bw/day.

 

Food Consumption

During the first week of treatment dietary intake was generally lower compared to controls at 75 and 150 mg/kg bw/day. However, improvement was evident thereafter.

No such effects were detected at 25 mg/kg bw/day.

 

Water Consumption

There was no effect evident on water intake at any dose level.

 

Necropsy

No macroscopic abnormalities were detected in the surviving females.

 

Litter Data and Placental/Fetal Weights

There were no obvious adverse effects of maternal treatment on litter data as assessed by the number of implantations, early and late embryonic/fetal deaths and live fetuses, or sex ratios as assessed by percentage males.

 

Fetal Examination

There were no treatment-related findings apparent for fetuses from treated females at external examination of Day 20 of gestation.

 

Conclusion

Based on the findings of this study, the administration of CGE-PMDA adduct at a dosage of 200 mg/kg bw/day is deemed too high for continued dosing. Although reduced body weight development was observed at 150 mg/kg bw/day with no reproductive effects to treatment, the effects were considered not to be significant enough to dismiss further investigation.

Therefore, dose levels of 25, 75 and 150 mg/kg bw/day are recommended for the forthcoming Oral (Gavage) Combined Repeat Dose Toxicity Study (with Recovery Groups) with Reproduction /Developmental Toxicity Screening Test in the Rat (OECD 422).