N,N,N',N'-tetrakis(2-hydroxypropyl)adipamide

Basic toxicokinetics

Administrative data

Endpoint:

basic toxicokinetics

Type of information:

experimental study

Adequacy of study:

key study

Study period:

from May 4, 2001 to July 1, 2002

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Remarks:

N, N, N’, N’-Tetrakis (2-hydroxyethyl) hexanediamide, which is structurally similar to N,N,N',N'-tetrakis(2-hydroxypropyl)adipamide. Thus, the guideline study with GLP is used for read-across to avoid duplicate tests.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2002

Report date:

2002

Materials and methods

Objective of study:

toxicokinetics

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 417 (Toxicokinetics)

Deviations:

not specified

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

PRIMID XL-552

IUPAC Name:

PRIMID XL-552

Test material form:

other: solid

Radiolabelling:

yes

Test animals

Species:

rat

Strain:

Wistar

Sex:

male/female

Details on test animals or test system and environmental conditions:

Animals
Species: Rat
Strain: HanBrl WIST: Wistar rats, outbred, SPF-quality
Rationale: Recognized by international guidelines as the recommended animal species representing the acceptable model in case of human exposure.
Source: RCC Ltd, Biotechnology and Animal Breeding Division Wolferstrasse 4, CH-4414 Fueliinsdorf/Switzerland
Initial number: 36 males and 36 females; additionally one reserve animal of each sex for each batch.
Actual number: 36 males and 36 females.
Body weights: The body weights were determined at the beginning of acclimation (body weight males: 140-160 g, females: 150- 170 g) and at the day of treatment with 14C-PRIMID XL-552 (body weight males and females: 135-172 g, groups 1-8). For groups 9 and 10, the body weight prior to the radiolabelled test item administration ranged from 191- 256 g.
Age at acclimation: Males 6-8 weeks, females: 7-10 weeks
Identification: Individual numbers (ear tags)
Acclimation: At least 5 days to laboratory environment, including 1-3 days to cages with a stainless steel grid or to ail-glass metabolism cages. The animals of groups 9 and 10 were placed into the metabolism cages immediately after the radiolabelled administration.
Allocation:
Randomization: Animal numbers will be given randomly.
Husbandry
Room: No. 132, air-conditioned
Conditions: Target temperature: 19.0 - 25.0 °C
Target relative humidity: 40.0 - 70.0%
Photocycle: 12 hours
Air changes: 10-15 times/hour

Accommodation:In groups of 2-4 rats in Makrolon cages (type 3) under conventional hygienic conditions with standard soft wooden bedding during acclimatisation. The accommodations during the treatments are described in the respective experiments.
Diet: Pelleted 3433-Kliba rat maintenance diet ad libitum * (PROV丨Ml KUBA AG, CH-4303 Kaiseraugst/ Switzerland). The rats were fasted overnight prior to administration.
Water: Tap water ad libitum
Health Status:The health status of the treated animals was checked visually at daily intervals.

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

corn oil

Details on exposure:

Prior to dosing the rats were fasted overnight (16-18 hours). Oral administration at a target volume of 1 ml/100 g rat body weight was performed by gastric intubation using an appropriate device. The actual amount of solution administered was determined by weighing the device before and after administration.

Duration and frequency of treatment / exposure:

- a balance study in both sexes at two dose levels after single oral administration (groups 1-4):
One treatment; Duration of recovery: 96 hours
- a blood and plasma level study in both sexes at two dose levels after single oral administration (groups 5-8): 24 males and 24 females
Target dose levels (mg)/kg) Time intervals (hours)
5.0 100 5.0 100
Group 5 6 7 8
Sex male male female female
Animal-no. 17-20 29-32 41-44 53-56 0, 2, 8, 72
21-24 33-36 45-48 57-60 0.5, 4, 24, 96
25-28 37-40 49-52 61-64 1, 6, 48
number of treatments: one treatment
duration of recovery: maximally 96 hours
- a balance study in both sexes after repeated oral administration of non-labelled test item (14x) followed by single oral administration of 14C-labelled test item at the low dose level (groups 9 and 10):
treatment: 14 daily administrations with unlabelled test item followed by one oral administration with the 14C-iabelled test item.
duration of recovery: 96 hours

Doses / concentrations

Remarks:

Doses / Concentrations:
Target dose level: Low dose ; 5 mg/kg of body weight (spec. rad. 13.5 jJCi/mg or 500 kBq/mg)
High dose: 100 mg/kg of body weight (spec. rad. 1,35 pCi or 50 kBq/mg)
Target oral
administration volume: 1.0 ml/100 g of body weight

No. of animals per sex per dose / concentration:

- a balance study in both sexes at two dose levels after single oral administration (groups 1-4):
step1: 4 males; step 2: 4 males and 8 females
- a blood and plasma level study in both sexes at two dose levels after single oral administration (groups 5-8): 24 males and 24 females
- a balance study in both sexes after repeated oral administration of non-labelled test item (14x) followed by single oral administration of 14C-labelled test item at the low dose level (groups 9 and 10): 4 males and 4 females

Control animals:

yes, concurrent vehicle

Positive control reference chemical:

no

Details on study design:

Levels of radioactivity in expired air, urine, faeces, blood, plasma and organs/tissues and the identity of metabolites in plasma, urine, faeces and organs/tissues were determined based on the following experiments:
- a balance study in both sexes at two dose levels after single oral administration (groups 1-4)
The study was performed in two steps. In the first study part, male rats were orally treated with the high dose level of 14C-PRIMID XL-552 and radioactivity was determined in expired air, urine, faeces, blood, organs/tissues, residual carcass and cage wash.
In the second part of the study , female rats were orally treated with the low and high dose levels and male rats were treated with the low dose level. Radioactivity was determined in urine, faeces, blood, organs/tissues, residual carcass and cage wash.

- a blood and plasma level study in both sexes at two dose levels after single oral administration (groups 5-8)

- a balance study in both sexes after repeated oral administration of non-labelled test item (14x) followed by single oral administration of 14C-labelled test item at the low dose level (groups 9 and 10)

Details on dosing and sampling:

- a balance study in both sexes at two dose levels after single oral administration (groups 1-4):
step 1: 100 mg/kg of body weight; sampliing: Volatiles, urine, faeces, Intestinal Tract/Carcass/Bones, Organs/Tissues/Blood/Plasma, Cage Wash
step 2: Target dose levels (mg/kg)
5.0 5.0 100
Group 2 3 4
Sex male female female
Animal-no. 5-8 9-12 13-16
sampling: urine, faeces, blood, organs/tissues, residua! carcass and cage wash
- a blood and plasma level study in both sexes at two dose levels after single oral administration (groups 5-8):
Group 5 and 7: 5.0 mg/kg of body weight (low dose)
Group 6 and 8: 100 mg/kg of body weight (high dose)
sampling:
Blood (approximately 0.5 ml) was withdrawn retroorbital from 4 individual animals at 0, 2 and 8 hours or 0.5, 4 and 24 hours or 1 and 6 hours after administration and collected into heparinized tubes. Aliquots were separated and used for determination of total radioactivity. The rest of the blood was centrifuged at about 1500-2000 g for 10 min. The decanted plasma and the blood pellets were stored at -20 °C.
Animals were sacrificed by carbon dioxide at 48, 72 and 96 hours after administration. Body weights were recorded. Blood was collected into heparinized tubes from the chest cavity after heart incision and worked up analogous to the retroorbital blood samples.

- a balance study in both sexes after repeated oral administration of non-labelled test item (14x) followed by single oral administration of 14C-labelled test item at the low dose level (groups 9 and 10):
5.0 mg/kg of body weight (group 9 and 10);
sampling: urine, faeces, blood, organs/tissues, residual carcass and cage wash.

Statistics:

For additional analyses (samples after pooling, protein precipitation, extraction), all values of radioactivity expressed in percentages as well as in parent equivalents on a fresh weight basis was compared to the average value initially calculated from the respective individual animals.
Areas under the mean blood-plasma-curves were calculated and relevant pharmacokinetic parameters were determined. The data were evaluated by Non-compartimental Pharmacokinetics Data Analysis using the software PK Solutions 2.0™ (Summit Research Services, Montrose, CO 81401 USA)

Results and discussion

Main ADME results

Main ADME results 1

Type:

absorption

Results:

Oral exposure is the expected route of exposure to humans.

Main ADME results 2

Type:

metabolism

Results:

AUC(0-∞) levels for blood and plasma ranged from 131.2-234.1 ng h/ml for the low dose and from 2035.1 to 4442.2 ng h/ml for the high dose.

Main ADME results 3

Type:

distribution

Results:

At 96 hours after the administration, radioactive residues in organs/tissues were very low.

Main ADME results 4

Type:

excretion

Results:

Excretion occurred in all groups rapidly mainly via faeces. Within 48 hours, the radioactivity was almost completely excreted. At 96 hours after the administration, radioactive residues in organs/tissues were very low.

Toxicokinetic / pharmacokinetic studies

Details on absorption:

Oral exposure is the expected route of exposure to humans.

Details on distribution in tissues:

At 96 hours after the administration, radioactive residues in organs/tissues were very low. After oral administration at the high dose (groups 1 and 4), organs tissues with the highest amounts were: blood (0.029-1.138 µg eq/g), carcass (0.008-0.011 µg eq/g), intestinal tract (0.036-0.045 (µg eq/g), liver (0.070-0.130 (µg eq/g)and skin (0.013-0.106 µg eq/g). For the low dose (groups 2, 3, 9 and 10) all values were below 0,01 µg eq/g.

Details on excretion:

Excretion occurred in all groups rapidly mainly via faeces. Within 48 hours, the radioactivity was almost completely excreted. At 96 hours after the administration, radioactive residues in organs/tissues were very low.

Toxicokinetic parameters

Toxicokinetic parameters:

half-life 1st: 2.7 - 4.7 hours

Metabolite characterisation studies

Metabolites identified:

yes

Details on metabolites:

Metabolites patterns were determined by HPLC and TLC analyses in the faeces and urine.
In faeces up to 6 radioactive fractions (F1-F6) were detected. Besides the parent item (70.0 - 83.2% of the radioactivity administered), the following metabolite fractions could be detected: F1 (<0.1 - 4.4%), F3 (2.4 - 3.5%), F4 (14 - 3.2%), and F5 (3.7 - 7.7%) and F6 (1.2 - 3.0 %).
In urine at least 4 radioactive fractions (U1-U4) were detected. The following metabolite fractions could be detected in the urine: U1 (parent, 2.8 - 3.5 %), U2 (<0.1 - 0.3 %), U3 (<0.1 - 0.4 %) and U4 (0.1 - In accordance with Column 2 of REACH, Annexes IX the tests do not need to be conducted based on exposure considerations: The study does not need to be conducted because direct or indirect exposure of the soil compartment is unlikely; the substance is used solely in industrial processes for powder coating, technically required being water-free and applied under closed conditions. Thus, there is no exposure to the soil or the environment during use and no additional studies are required.0.4%).

Any other information on results incl. tables

The following balance was obtained (group means in percent of the radioactivity administered):

	Time interval (hours)	Animal group
		1	2	3	4	9	10
Urine	0-8	1.94	2.90	2.55	3.15	2.72	2.10
	8-24	2.06	1.02	0.75	0.66	1.07	0.66
	24-48	0.11	0.28	0.24	0.13	0.17	0.10
	48-72	0.04	0.05	0.16	0.15	0.04	0.05
	72-96	0.04	0.02	0.17	0.07	0.01	0.05
Subtotal		4.19	4.27	3.86	4.16	4.00	2.96
Faeces	0-24	77.06	83.49	72.6	89.2	86.45	78.81
	24-48	15.75	12.79	19.78	9.61	11,10	13.55
	48-72	1.11	0.94	3.15	0.95	0.67	3.12
	72-96	0.3	0.23	0.35	0.2	0.23	1.18
Subtotal		94.22	97.45	95.87	99.95	98.45	96.66
Cage wash		1.25	1.11	2.93	1.13	0.48	0.98
Volatiles	0-96	0.73	—	—	—	一
Total excreted		100.39	102.83	102.67	105.24	102.93	100.59
Kidney		<0.01	<0.01	<0.01	<0.01	<0.01	<0.01
Liver		0.01	0.01	< 0.01	<0.01	<0.01	<0.01
Organs/tissues		0.09	<0.01	<0.01	<0.01	<0.01	<0.01
Intestinal tract		0.01	<0.01	0.01	0.01	0.01	<0.01
Carcass		0.01	<0.01	0.01	0.01	<0.01	0.04
Subtotal		0.11	0.01	0.01	0.02	0.01	0.04
Total		100.50	102.83	102.68	105.25	102.94	100.63

The following organs/tissues levels were measured (group means in pg eq/g):

	1	2	Anima 3	group 4	9	10
Adrenal gland*	0.000	0.000	0.000	0.000	0.000	0.000
Blood	1.138	0.001	0.001	0.029	0.000	0.000
Brain	0.038	0.000	0.000	0.000	0.000	0.000
Carcass	0.011	0.000	0.001	0.008	0.001	0.003
Epididymes	0.010	0.000		—	0.000	—
Fat	0.047	0.000	0.000	0.000	0.000	0.000
Femur	0.003	0.000	0.001	0.011	0.000	0.000
Heart	0.037	0.000	0.000	0.000	0.000	0.000
Intestinal tract	0.045	0.000	0.003	0.036	0.003	0.003
Kidney	0.028	0.000	0.000	0.003	0.000	0.000
Liver	0.130	0.006	0.001	0.070	0.000	0.000
Lung	0.017	0.000	0.000	0.000	0.000	0.000
Muscie	0.028	0.002	0.000	0.000	0.000	0.000
Ovary*	—-	—	0.000	0.000	--	0.000
Pancreas	0.058	0.000	0.000	0.000	0.000	0.000
Plasma	0.239	0.001	0.001	0.003	0.000	0.000
Skin	0.106	0.000	0.001	0.013	0.000	0.000
Spleen	0.045	0.000	0.000	0.000	0.000	0.000
Stomach	0.043	0.000	0.000	0.000	0.000	0.004
Testicles	0.044	0.000	一-	—	0.000	___
Thymus	0.035	0.000	0.000	0.000	0.000	0.000
Thyroid gland *	0.008	0.000	0.000	0.000	0.000	0.000
Uterus	…	—	0.001	0.006	…	0.001

After orai administration of [¹⁴C]-PRIMID XL-552, excretion of radioactivity occurred rapidly, mainly via faeces, almost completely within 48 hours. No differences in the excretion patterns between male/female rats, low/fiigh dose as well as single/repeated administration could be observed.

In conclusion, after oral administration of¹⁴C-PRIMID XL-552 to male and female rats at two dose levels, generally very low levels of radioactive residues in organs/tissues were measured.

After single oral administration of¹⁴C-PRIMID XL-552 at the high dose level, higher values were found in the organs/tissues of group 1 (males, high dose) as compared to group 4 (females, high dose). However, no sex related difference was found for the low dose level after single and repeated oral administration.

Applicant's summary and conclusion

Conclusions:

Interpretation of results: No bioaccumulation potential was observed in study results; within 48 hours the radioactivity was almost completely excreted.
Based on the results obtained after single oral administration of [14C]-test article to male and female rats at two dose levels as well as after repeated oral administration of unlabelled PRIMID followed by [14C]-PRIMID XL-552 at the low dose level to male and female rats, the following can be concluded:
Excretion occurred in all groups rapidly, mainly via faeces. Within 48 hours, the radioactivity was almost completely excreted. At 96 hours after the administration, radioactive residues in organs/tissues were very low.
Concerning pharmacokinetic parameters, no differences between male/female rats, low/high dose as well as single/repeated administration were found.
The major amount of the radioactivity in the faeces could be extracted by using acetonitrile/purified water (8+2, v/v). In faeces up to 6 radioactive fractions (F1-F6) were detected. F2 was identical to the parent item. Fractions F1, F3-F6 were unknown metabolite fractions.
In urine at least 4 radioactive fractions (U1-U4) were detected. U1 was identical to the parent item. Fractions U2-U4 were unknown metabolite fractions.

Executive summary:

This study was conducted in compliance with OECD guideline 417 and GLP principles with both sexes exposed to test item at low dose level and high dose level of 5 mg/kg/day and 100 mg/kg/day, respectively. The object of this study was to follow the absorption, distribution, metabolism and excretion of 14C-test article after single and repeated oral administration to rats.

Excretion occurred in all groups rapidly, mainly via faeces. Within 48 hours, the radioactivity was almost completely excreted. At 96 hours following administration, radioactive residues in organs/tissues were very low. Concerning pharmacokinetic parameters, no differences between male/female rats, low/high dose as well as single/repeated administration were found. The major amount of the radioactivity in the faeces could be extracted by using acetonitrile/purified water (8+2, v/v). In faeces up to 6 radioactive fractions (F1-F6) were detected. F2 was identical to the parent item. Fractions F1, F3-F6 were unknown metabolite fractions. In urine at least 4 radioactive fractions (U1-U4) were detected. U1 was identical to the parent item. Fractions U2-U4 were unknown metabolite fractions.