Amines, N-(3-aminopropyl)-N-(C16-18 and C18-unsatd. alkyl)trimethylenedi-

Basic toxicokinetics

Administrative data

Endpoint:

basic toxicokinetics

Type of information:

experimental study

Adequacy of study:

key study

Study period:

17-01-1996 to 24-04-1996

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP-compliant guideline study, no restrictions, fully adequate for assessment.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1996

Materials and methods

Objective of study:

toxicokinetics

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 417 (Toxicokinetics)

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

N-(3-aminopropyl)-N-dodecylpropane-1,3-diamine

EC Number:

219-145-8

EC Name:

N-(3-aminopropyl)-N-dodecylpropane-1,3-diamine

Cas Number:

2372-82-9

Molecular formula:

C18H41N3

IUPAC Name:

N-(3-aminopropyl)-N-dodecylpropane-1,3-diamine

Details on test material:

- Name of test material (as cited in study report): P4198
N,N-bis-(3-aminopropyl)dodecylamine
- Physical state: Liquid
- Radiochemical purity (if radiolabelling): ≥96%
- Specific activity (if radiolabelling): 19.7mCi/mmol; 65.8µCi/mg

Radiolabelling:

yes

Remarks:

14C

Test animals

Species:

rat

Strain:

Sprague-Dawley

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Harlan UK, Bicester, Oxfordshire, UK
- Age at study initiation: ca. 6-10 weeks
- Weight at study initiation: ca. 200 g
- Fasting period before study:
- Housing: during acclimatisation in stainless steel cages with suspended mesh floor. Rats from excretion study were housed in individual glass metabolism cages, rats from other studies (blood/plasma kinetics and tissue distribution) in stainless steel battery cages (25 x 25 x 50 cm).
- Individual metabolism cages: yes
- Diet (e.g. ad libitum): Standard Laboratory Rat LAD 1, ad libitum
- Water: ad libitum
- Acclimation period: at least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19-23
- Humidity (%): 40-60
- Photoperiod (hrs dark / hrs light): 12 / 12

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

unchanged (no vehicle)

Duration and frequency of treatment / exposure:

single dose

Doses / concentrations

Remarks:

Doses / Concentrations:
3 and 30 mg/kg bw

No. of animals per sex per dose / concentration:

Excretion studies:
Eight rats (4 male, 4 female) received single oral doses of 14C-P4198 at a nominal dose level of 3 mg/kg bw
Eight rats (4 male, 4 female) received single oral doses of 14C-P4198 at a nominal dose level of 30 mg/kg bw
Blood/plasma kinetics: Eighteen animals (9 male, 9 female) received single oral doses (nominal 3 mg/kg bw) of 14C-P4198
Quantitative tissue distribution: five male rats received single oral doses (nominal 3 mg/kg bw) of 14C-P4198

Control animals:

no

Details on study design:

- Dose selection rationale: the high dose of 30 mg/kg bw was selected as ca. 10-fold lower dose of LD50 (261 mg/kg bw). A further 10-fold lower dose was selected as a dose at which no toxic effects were expected.

Details on dosing and sampling:

PHARMACOKINETIC STUDY (Absorption, distribution, excretion)
- Tissues and body fluids sampled: urine, faeces, expired air, blood, cage wash
- Time and frequency of sampling: urine was collected at 0-6 and 6024 hours after dosing and then at 24 hours interval up to 120 hours. Faeces were collected at 24 hours interval for 120 hours. Expired air was collected at 24 hours interval for 48 hours. Cages were washed at 120 hours.

BLOOD/PLASMA kinetics:
- Time and frequency of sampling: Group 1: Pre-dose, 14, 24 and 96 hours. Group 2: 0.25, 2, 6, 48 and 120 hours. Group 3: 0.5, 3, 12, 72 and 168 hours.

QUALITATIVE TISSUE DISTRIBUTION:
- Time and frequency of sampling: at 3, 6, 24, 48 and 120 hours post-administration sacrifce

METABOLITE CHARACTERISATION STUDIES:
- Tissues and body fluids sampled: urine, faeces, expired air, blood, cage wash, tissues
- Time and frequency of sampling: urine was collected at 0-6 and 6024 hours after dosing and then at 24 hours interval up to 120 hours. Faeces were collected at 24 hours interval for 120 hours. Expired air was collected at 24 hours interval for 48 hours. Cages were washed at 120 hours. Blood was taken from a tail vein at following intervals: Group 1: Pre-dose, 14, 24 and 96 hours. Group 2: 0.25, 2, 6, 48 and 120 hours. Group 3: 0.5, 3, 12, 72 and 168 hours. Tissues were analyzed at at 3, 6, 24, 48 and 120 hours post-administration sacrifce.
- From how many animals: (samples pooled or not): 2 groups (2 dose levels) of 8 rats (4 males and 4 females) for excretion studies; 6 groups (2 dose levels, 3 groups each) of 6 rats (3 males and 3 females) for blood/plasma kinetics studies; 5 males for qualitative tissue distribution studies. Samples were pooled.
- Method type(s) for identification: liquid scintillation counting; TLC for samples of faecal extracts.

Results and discussion

Preliminary studies:

No visible toxic effects were observed in the 4 rats that received a single oral dose of 30 mg/kg of non-radiolabelled P4198. The high dose was confirmed as 30 mg/kg.

Main ADME results

Type:

absorption

Results:

The gastrointestinal absorption is considered to be 2.5% based on a retained dose.

Toxicokinetic / pharmacokinetic studies

Details on absorption:

Low level doses:
After single low level doses of 14C-P4198 to three groups of six rats (3 male, 3 female), peak of mean concentrations of radioactivity in plasma were below the limit of reliable measurement. Peak plasma concentration occurred at 4 hours after administration of the test substance.
High level doses:
After single high level doses of 14C-P4198 to three groups of six rats (3 male, 3 female), peak of mean concentrations of radioactivity in plasma of 0.12 (male) and 0.12 (female) µg equiv./ml occurred at 12 hours. Concentrations declined with an approximate half-life (measured between 12 and 48 hours) of 28.5 (male) and 46.3 (female) hours. The area under the concentration v time curve (AUCt) was 4.6 (male) and 5.1 (female) µg equiv./ml.hour.

Details on distribution in tissues:

At 3 hours after administration of the 14C-test compound at a nominal dose level of 3 mg/kg, high concentrations of radioactivity were detected in the oesophagus as well as in the contents of the caecum, small intestine and stomach. Intermediate concentrations of radioactivity were detected in the bladder. Lowest concentrations of radioactivity were detected in the adrenals, blood, kidney, large intestine contents, liver, lungs, pancreas, salivary gland and spleen.
At 6 hours after administration, high concentrations of radioactivity were detected in the oesophagus as well as in the contents of the caecum, small intestine and stomach. Intermediate concentrations of radioactivity were detected in the kidney, liver, lungs and spleen. Lowest levels of radioactivity were detected in the adrenals, brown fat, blood, bone marrow, exorbital lachrymal gland, myocardium, pancreas, preputial gland, pituitary gland, salivary gland, thymus and thyroid.
At 24 hours after administration, high concentrations of radioactivity were detected in the contents of the caecum, large intestine and stomach. Intermediate concentrations of radioactivity were detected in the adrenals, brown fat, Harderian gland, kidney, liver, lung, myocardium, preputial gland, oesophagus, small intestine contents, salivary gland and spleen. Lowest levels of radioactivity were in the blood, bone marrow, exorbital lachrymal gland, intra-orbital lachrymal gland, pancreas, pituitary, thymus and thyroid.
At 48 hours after administration, high levels of radioactivity were detected in the contents of the caecum, the large intestine and the large intestine mucosa. Intermediate concentrations of radioactivity were detected in the kidney, lung, small intestine contents, small intestine mucosa, spleen, stomach contents and stomach mucosa. Lowest concentrations of radioactivity were detectd in the adrenals, blood, bone marrow, exorbital lachrymal gland, Harderian gland, intra-orbital lachrymal gland, liver, myocardium, pancreas, preputial gland, pituitary gland, salivary gland and thymus.
At 120 hours after administration, intermediate concentrations of radioactivity were detected in the kidney, lung, pancreas, salivary gland, smal intestine mucosa, spleen and stomach mucosa. Lowest levels of radioactivity were detected in the adrenals, blood, bone marrow, exorbital lachrymal gland, Harderian gland, liver, intra-orbital lachrymal gland, muscle, myocardium, pituitary gland, thymus, testes and thyroid.

Details on excretion:

Low level dose:
After a single low level oral dose of 14C-P4198 to rats (4 male, 4 female), means of 0.2% (male) and 0.2% (female) were excreted in the urine during 0 – 120 hours. Most of this radioactivity was excreted in the 0 – 48 hour urine (0.1% for both male and female). During the 5 days after dosing 97% (male) and 96% (female) was excreted in the faeces with most of this in the 0 – 72 hour samples (97% male, 95% female). Radioactivity in expired air traps was measured over 48 hours and accounted for 0.1% (male) and 0.2% (female) dose. After sacrifice at 120 hours radioactivity in the gastrointestinal tract accounted for 0.5% (male) and 0.4% (female) dose whilst radioactivity in the remaining carcass accounted for 1.0% (male) and 1.5% (female) dose. Thus, means of 99% (male) and 98% (female) were recovered after 5 days.
High level dose:
After a single high level oral dose of 14C-P4198 to rats (4 male, 4 female), means of 0.2% (male) and 0.2% (female) were excreted in the urine during 0 – 120 hours. Most of this radioactivity was excreted in the 0 – 48 hour urine (0.2% for male and 0.1% for female). During the 5 days after dosing 95% (male) and 90% (female dose) was excreted in the faeces with most of this in the 0 – 72 hour samples (95% male, 89% female). Radioactivity in expired air traps was measured over 48 hours and accounted for 0.4% (male) and 0.4% (female) dose. After sacrifice at 120 hours radioactivity in the gastrointestinal tract accounted for 1.2% (male) and 1.3% (female) dose whilst radioactivity in the remaining carcass accounted for 3.0% (male) and 2.6% (female) dose. Thus, means of 100% (male) and 94% (female) were recovered after 5 days.

Metabolite characterisation studies

Metabolites identified:

yes

Details on metabolites:

Metabolites were quantified using TLC. Samples of faecal extracts were analysed directly. Urine was not analysed as the total radioactivity excreted in the urine was < 0.3%.
Faecal extracts:
In the 0 – 48 hour faecal extracts of rats after administration of 14C-P4198 there was one major radioactive component that accounted for 7 – 13% dose (39 – 51% sample radioactivity). This component co-chromatographed with authentic P4198 in two TLC systems. Six further minor components were detected in faecal extracts accounting for 1 –3% dose after both low and high level doses. One of these components was shown to co-chromatograph with 1-dodecylamine using TLC. The nature of the other components was not investigated further due to the low levels of radioactivity involved (<3% dose).
A large proportion of the dose (66 – 72%) remained unextracted after both low and high level doses. Further extraction with 1M HCl and 2M NaOH extracted a further 17 – 20% (low level) and 20 –25% (high level) dose. Attempts to analyses these extracts (after neutralisation) were unsuccessful.

Any other information on results incl. tables

After single oral doses of 14C-P4 198 approximately 0.2% of the dose was excreted in the urine. Approximately 90-97%
dose was excreted in faeces during 5 days. At sacrifice approximately 1 -3% dose was retained in the carcass and 0.4
- 1.3 % of the dose was recovered in the gastrointestinal tract. Radioactivity in expired air traps accounted for 
0.4% dose. There was no noticeable difference in excretion after the different dosing regimens or between male and
female rats. This indicated that an average of 2.5% of the initial dose was not excreted.

Concentrations in plasma were consistent with low absorption of P4198 from the gastrointestinal tract. After low level
doses, concentrations were generally below the limit of reliable measurement and appeared to peak 4 hours after dose
administration. After high level doses, concentrations of radioactivity in plasma were again low with values generally
less than twice the limit of reliable measurement. The plasma concentrations of radioactivity indicated a fairly
rapid clearance of radioactivity with peak concentrations measured 12 hours after dosing. Qualitative analysis of the
tissue distribution of radioactivity by whole- body autoradiography indicated high levels of radioactivity in
the gastrointestinal tract and its contents. Levels of radioactivity observed in other tissues were consistent with
low absorption from the gastrointestinal tract.
The major component in faecal extracts was unchanged P4198, accounting for 7-13% dose. The assignment was confirmed by
co-chromatography using TLC. Six minor components were also observed in faecal extracts, each accounting for 1 -3 %
dose. One of these was shown to co-chromatograph with 1-dodecylamine.
As the liver showed constant less activity then the kidney in all  radiographs, significant entero-hepatic circulation can be excluded

Applicant's summary and conclusion

Executive summary:

After single oral doses of 14C-P4 198 approximately 0.2% of the dose was excreted in the urine. Approximately 90-97% dose was excreted in faeces during 5 days. At sacrifice approximately 1 -3% dose was retained in the carcass and 0.4 - 1.3 % of the dose was recovered in the gastrointestinal tract. Radioactivity in expired air traps accounted for 0.4% dose. There was no noticeable difference in excretion after the different dosing regimens or between male and female rats. This indicated that an average of 2.5% of the initial dose was not excreted.

 

Concentrations in plasma were consistent with low absorption of P4198 from the gastrointestinal tract. After low level doses, concentrations were generally below the limit of reliable measurement and appeared to peak 4 hours after dose administration. After high level doses, concentrations of radioactivity in plasma were again low with values generally less than twice the limit of reliable measurement. The plasma concentrations of radioactivity indicated a fairly rapid clearance of radioactivity with peak concentrations measured 12 hours after dosing. Qualitative analysis of the tissue distribution of radioactivity by whole- body autoradiography indicated high levels of radioactivity in the gastrointestinal tract and its contents. Levels of radioactivity observed in other tissues were consistent with low absorption from the gastrointestinal tract.

The major component in faecal extracts was unchanged P4198, accounting for 7-13% dose. The assignment was confirmed by co-chromatography using TLC. Six minor components were also observed in faecal extracts, each accounting for 1 -3 % dose. One of these was shown to co-chromatograph with 1-dodecylamine.

As the liver showed constant less activity then the kidney in all radiographs, significant entero-hepatic circulation can be excluded.