Registration Dossier

Administrative data

Description of key information

Only insufficient data with Amines, polyethylenepoly-, triethylenetetramine fraction are available. A 90 -day study according to OECD 408 inlcuding a developmental/reproduction screening is ongoing. As soon as the data is available the dossier will be updated. Therefore, studies with the structural analogue substance TETA-2HCl (CAS 38260 -01 -4) are used as interim approach and considered reliable to evaluate the repeated dose toxicity of Amines, polyethylenepoly-, triethylenetetramine fraction.

LOAEL (TETA-2HCl, rat, oral, subchronic) = 50 mg/kg bw/day; recalculation for Amines, polyethylenepoly-, triethylenetetramine fraction based on molecular weight results in a LOAEL of 33 mg/kg bw/day

Key value for chemical safety assessment

Repeated dose toxicity: via oral route - systemic effects

Link to relevant study records
Reference
Endpoint:
sub-chronic toxicity: oral
Type of information:
read-across from supporting substance (structural analogue or surrogate)
Remarks:
Summary of available data used for the endpoint assessment of the target substance
Adequacy of study:
key study
Justification for type of information:
Considering the target substance Amines, polyethylenepoly-, triethylenetetramine fraction (EC no.: 292-588-2, CAS no.: 90640-67-8) hereafter referred to as TETA, the following source substance has been considered for the read-across: N,N'-bis(2-aminoethyl)ethylenediamine dihydrochloride (EC no.: 253-854-3, CAS no.: 38260-01-4) known as well as Trientine x 2HCl, hereafter referred to as TETA x 2HCl. It has to be noted that the target substance is a multi-constituent substance and shares the same main constituent with TETA x 2HCl but in form of an ammonium hydrochloride salt. TETA x 2HCl is considered to be a mono-constituent substance. The source substance (TETA x 2HCl) is not completely characterized with regards to the presence of impurities. The purity degree, as stated in its MSDS, suggests it is a highly pure mono-constituent substance (TETA x 2 HCl up to 100% pure). Therefore, taking into account the latter mentioned deficiency in the read-across approach, the registrants agree to conduct an OECD 408 study on the registration substance that is also in line with the request in the draft decision received from ECHA on 13 December 2018 (DEV-01-2119487919-13-0000-CCH-2_DEC_REG_CCH-D-2114449865-34-01). Until the OCED 408 study data on the registered substance are available, the data on the source substance TETA x 2 HCl will be used as interim approach to conduct quantitative risk assessment.
Reason / purpose for cross-reference:
read-across source
Reason / purpose for cross-reference:
read-across source
Reason / purpose for cross-reference:
read-across source
Dose descriptor:
NOAEL
Remarks:
rat
Effect level:
50 mg/kg bw/day (actual dose received)
Based on:
test mat.
Sex:
female
Basis for effect level:
other: irreversible toxic changes in the lung
Remarks on result:
other: CAS 38260-01-4, Yanagisawa, 1998
Dose descriptor:
LOAEL
Remarks:
rat
Effect level:
50 mg/kg bw/day (actual dose received)
Based on:
test mat.
Sex:
male
Basis for effect level:
other: irreversible toxic changes in the lung
Remarks on result:
other: CAS 38260-01-4, Yanagisawa, 1998
Dose descriptor:
NOAEL
Remarks:
dog
Effect level:
50 mg/kg bw/day (actual dose received)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
neuropathology
Remarks on result:
other: CAS 38260-01-4, Maemura, 1998
Dose descriptor:
NOAEL
Remarks:
mice
Effect level:
600 ppm
Based on:
test mat.
Remarks:
corresponds to 92 mg/kg bw
Sex:
male/female
Basis for effect level:
other: see remarks
Remarks on result:
other: CAS 38260-01-4, Greenman, 1996
Dose descriptor:
NOAEL
Remarks:
rats
Effect level:
>= 3 000 ppm
Based on:
test mat.
Remarks:
3000 ppm corresponds to 270 mg/kg bw
Sex:
male
Basis for effect level:
other: No test substance-related effects observed.
Remarks on result:
other: CAS 38260-01-4, Greenman, 1996
Dose descriptor:
NOAEL
Remarks:
rats
Effect level:
600 ppm
Based on:
test mat.
Remarks:
600 ppm corresponds to 60 mg/kg bw/day
Sex:
female
Basis for effect level:
other: uterine
Remarks on result:
other: CAS 38260-01-4, Greenman, 1996
Critical effects observed:
yes
Lowest effective dose / conc.:
50 mg/kg bw/day (actual dose received)
System:
respiratory system: lower respiratory tract
Organ:
lungs
Treatment related:
yes
Dose response relationship:
not specified
Relevant for humans:
not specified
Conclusions:
A 90-day study is availabe with the source substance triethylenetetramine dihydrochloride in mice and rats. Due to inflammation of the lung interstitium, hematopoietic cell proliferation of the spleen, liver periportal fatty infiltration, kidney weight reduction, reduced renal cytoplasmic vacuolization, and body weight gain reduction the NOAEL for mice was set to 92 mg/kg bw/day. In male rats no substance-related effects were observed and in female rats changes in the uterus were observed resulting in NOAEL (male) of 270 mg/kg bw/day and NOAEL (female) = 60 mg/kg bw/day. Sub-chronic treatment (26-weeks) of rats with the source substance triethylenetetramine dihydrochloride was associated with death and irreversible toxic changes in the lung. A dosage of 50 mg/kg bw/day was considered to be the NOAEL for females and for males the NOAEL was considered to be less than 50 mg/kg/day. Sub-chronic treatment (26-weeks) of dogs with the source substance triethylenetetramine dihydrochloride was associated with musculoskeletal effects. A dosage of 50 mg/kg bw/day was considered to be the NOAEL for female and male dogs. As explained in the analogue justification, it is considered that the target and the source substances are unlikely to lead to differences in repeated dose toxicity.
Endpoint conclusion
Endpoint conclusion:
adverse effect observed
Dose descriptor:
LOAEL
33 mg/kg bw/day
Study duration:
subchronic
Species:
rat
Quality of whole database:
The available information comprises adequate, reliable and consistent studies, from a reference substances with similar structure and intrinsic properties. Read-across is justified based on similarities in PC/ECO/TOX properties. The selected study is thus sufficient to fulfil the standard information requirements set out in Annex VIII-IX, 8.6, in accordance with Annex XI, 1.5, of Regulation (EC) No 1907/2006.
System:
respiratory system: lower respiratory tract
Organ:
lungs

Repeated dose toxicity: inhalation - systemic effects

Endpoint conclusion
Endpoint conclusion:
no study available

Repeated dose toxicity: inhalation - local effects

Endpoint conclusion
Endpoint conclusion:
no study available

Repeated dose toxicity: dermal - systemic effects

Endpoint conclusion
Endpoint conclusion:
no study available

Repeated dose toxicity: dermal - local effects

Endpoint conclusion
Endpoint conclusion:
no study available

Additional information

There are only insufficient data available on repeated dose toxicity with Amines, polyethylenepoly-, triethylenetetramine fraction. In order to fulfil the standard information requirements set out in Annex VII-IX, 8.6., in accordance with Annex XI, 1.5, of Regulation (EC) No 1907/2006, read-across from a structurally related substance was conducted. In accordance with Article 13 (1) of Regulation (EC) No 1907/2006, "information on intrinsic properties of substances may be generated by means other than tests, provided that the conditions set out in Annex XI are met.” In particular for human toxicity, information shall be generated whenever possible by means other than vertebrate animal tests, which includes the use of information from structurally related substances (grouping or read-across). Having regard to the general rules for grouping of substances and read-across approach laid down in Annex XI, Item 1.5, of Regulation (EC) No 1907/2006 whereby substances may be predicted as similar provided that their physicochemical, toxicological and ecotoxicological properties are likely to be similar or follow a regular pattern as a result of structural similarity.

Triethylenetetramine dihydrochloride (CAS 38260-01-4) and Amines, polyethylenepoly-, triethylenetetramine fraction (CAS 90640 -67 -8) are considered to be similar on the basis of the structural similar properties and/or activities. It has to be noted that the target substance is a multi-constituent substance and shares the same main constituent with TETA x 2HCl but in form of an ammonium hydrochloride salt. TETA x 2HCl is considered to be a mono-constituent substance. The source substance (TETA x 2HCl) is not completely characterized with regards to the presence of impurities. The purity degree, as stated in its MSDS, suggests however highly pure mono-constituent substance (TETA x 2 HCl up to 100% pure). Therefore, taking into account the latter mentioned deficiency in the read-across approach, the registrants agree to conduct an OECD 408 study on the registration substance that is also in line with the request in the final decision received from ECHA on 2 September 2019 (CCH-D-2114482145 -49 -01/F). Until the OCED 408 study data on the registered substance are available, the data on the source substance TETA x 2 HCl will be used as interim approach to conduct quantitative risk assessment.

 

Short-term toxicity

A key study similar to OECD 407 with the structural analogue substance TETA-2HCl is available (Yanagisawa, 1998). Triethylenetetramine dihydrochloride (TETA-2HCl), a copper chelating agent used to treat Wilson's disease, was administered orally to male and female F-344 rats for 4 or 8 weeks at dosages of 0, 100, 350 or 1200 mg/kg bw/day. Two males receiving 1200 mg/kg bw/day died during week 8 of treatment. In males receiving 1200 mg/kg bw/day during weeks 5 to 8 of treatment, body weight gain and food consumption were decreased and hunched posture and thin build were observed. During week 4 or 8 of treatment urinalysis revealed, for males receiving 100 mg/kg bw/day or animals receiving 350 mg/kg bw/day or more, increased electrolyte outputs possibly due to the hydrochloride nature of TETA-2HCl, with low plasma alkaline phosphatase activities evident in animals receiving 350 or 1200 mg/kg bw/day. After 4 and 8 weeks, and during 8 weeks of treatment, high lung weights and bronchiolar epithelium hypertrophy and broncho-alveolar pneumonia were recorded for animals receiving 1200 mg/kg bw/day, and submucosal acute inflammation within the glandular region of the stomach was recorded for males receiving 350 or 1200 mg/kg bw/day and in all treated female groups. Those effects lead to a systemic NOAEL of 350 mg/kg bw/day.

 

A 7 -day repeated dose toxicity study with Amines, polyethylenepoly-, triethylenetetramine fraction is available which is used as supporting study (Meyers, 1976). The test substance was added to the diet and fed to groups of 5 Wistar rats (30 days of age) per sex for 7 days. In the first part of the study 4 groups were used, these were dosed at 0, 0.55, 1.35 and 3.35 g/kg bw/day. Dosages attained (calculated based on nominal levels and food intake) were: 0, 0.51, 1.23, 2.98 g/kg bw/day for males, and 0, 0.47, 1.38, 2.63 g/kg bw/day for females. Animals treated at the highest dose and second highest dose showed a decrease in food intake, body weight loss, and decreased absolute and relative liver weight and decreased relative kidney weight. The NOAELs, therefore, were, 0.51 and 0.47 g/kg bw/day for males and females, respectively.

 

Sub-chronic toxicity

A key study similar to OECD 408 with the structural analogue substance TETA.2HCl is available (Greenman, 1996). Mice and rats received TETA.2HCl in the drinking water at concentrations of 0, 120, 600, or 3000 ppm for up to 92 days; they were fed diets containing nutritionally adequate levels of copper. An additional control group of rats and mice received a Cu-deficient diet. This low copper diet resulted in Cu-deficiency symptoms, such as anemia, liver periportal cytomegaly, pancreatic atrophy and multifocal necrosis, spleen hematopoietic cell proliferation, and increased heart weight, together with undetectable levels of plasma copper in rats but not in mice. TETA.2HCl lowered plasma copper levels somewhat (at 600 and 3000 ppm) in rats, but did not induce the usual signs of copper deficiency. TETA.2HCl caused an increased frequency of uterine dilatation at 3000 ppm in rats fed the adequate copper diet but was not noted in females fed the Cu-deficient diet. This observation may reflect an altered endocrine milieu since uterine dilatation is a normal phenomenon of the estrous cycle and is known to be sensitive to changes in estrogen exposure. TETA.2HCl toxicity occurred only in mice in the highest dose group. Increased frequencies of inflammation of the lung interstitium and liver periportal fatty infiltration were seen in both sexes, and hematopoietic cell proliferation was seen in the spleen of males. Kidney and body weights were reduced in males as was the incidence of renal cytoplasmic vacuolization. There were no signs of copper deficiency in mice exposed to TETA.2HCl. Based on the effects in mice the NOAEL was 600 ppm or 92 mg/kg bw/day for males and 99 mg/kg bw/day for females. In rats the NOAEL was 3000 ppm or 270 mg/kg bw/day for males and 600 ppm or 60 mg/kg bw/day for females.

 

A further study similar to OECD 408 with the structural analogue substance TETA.2HCl is available (Yanagisawa, 1998). Triethylenetetramine dihydrochloride (TETA.2HCl), a copper chelating agent used to treat Wilson's disease, was administered orally to male and female F-344 rats for 26 weeks at dosages of 0, 50, 175 or 600 mg/kg bw/day. One male receiving 175 mg/kg bw/day and three males receiving 600 mg/kg bw/day died, showing lung changes. With regard to the lungs, histopathology revealed a dose-related incidence and severity of focal chronic interstitial pneumonitis accompanied by fibrosis of the alveolar walls in females receiving 175 mg/kg/day or more and all treated male groups. Apart from the histological changes found in the lung, all other changes were reversible. It was concluded that the NOAEL of TETA.2HCl in this 26 -week study was considered to be 50 mg/kg bw/day for females and less than 50 mg/kg bw/day for males; the latter, therefore, was considered a LOAEL.

The authors of the study assume that TETA.2HCl could be taken up into the bronchiolar epithelium and/or alveolar epithelium by an endogenous polyamine uptake process and therefore could accumulated in the specific tissue site of the lung. Polyamines are generally irritating agents for mucous membranes, upper respiratory tract and skin (Maisey et al., 1988). In the 4 - or 8 -week study, the histopathological changes observed in the stomach (submucosal inflammation within the glandular region) indicates that TETA.2HCl was slightly irritating. Therefore, it is likely that the chronic interstitial pneumonitis was caused by the cytotoxic effect of TETA.2HCl which could have accumulated in the bronchiolar epithelial cells and the alveolar pneumocytes.

 

In addition, a subchronic study in dogs similar to OECD 409 is available as supporting study (Maemura, 1998). Beagle dogs received TETA.2HCl orally at dosages of 50, 100 or 200 mg/kg bw/day for 26 weeks followed by a 13 -week reversibility phase. However, in view of the severe signs that resulted in the sacrifice for human reasons of two males and one female receiving 200 mg/kg bw/day during week 9 of treatment, surviving dogs of this group were only treated for 10 weeks. Signs before killing included marked underactivity, body tremors, abnormal gait, limited use of limbs and prone posture. The ante mortem neurological examination generally indicated depressed postural and flexor withdrawal reactions. The signs were rapidly reversible except in one female which was killed humanly on day 2 of the reversibility period. Abnormal "stiff legged" gait and underactivity were evident, from week 23 of treatment, in two males and one female receiving 100 mg/kg bw/day. In the absence of any macroscopic or histopathologic findings, even after the examination of additional samples of muscle and nerve, the exact nature of this condition could not be elucidated. In all treated groups low copper and zinc concentrations in the livers and high urinary copper and zinc concentrations were found. The NOAEL was considered to be 50 mg/kg bw/day.

 

Overall, the results from the 26 -week key study in rats (Yanagisawa, 1998) and the 13 -week study in mice (Greenman, 1996) showed focal chronic interstitial pneumonitis. In the 13 -week study in rats described by Greenman, 1996 female rats resulted effects in the uterine only. The 26 -week study in dogs (Maemura, 1998) did not show lung toxicity but showed neurological effects instead. The NOAELs in these studies were in the same order of magnitude, i.e. between 50 and 99 mg/kg bw/day, although for male rats the level of 50 mg/kg bw/day should be considered a LOAEL.

 

The above study results with the structural analogue substance triethylenetetramine dihydrochloride (CAS 38260-01-4) indicate that the registration substance (CAS 90640 -67 -8) could also result in adverse effects after repeated oral exposure. To confirm whether the registration substance will cause adverse systemic effects after repeated oral exposure, an OECD 408 study on the registration will be conducted. The level of 50 mg TETA.2HCl per kg bw per day corresponds to (146.23 g/mol / 219.16 g/mol) x 50 mg/kg bw/day = 33 mg Amines, polyethylenepoly-, triethylenetetramine fraction per kg bw per day. Thus, the LOAEL of 33 mg/kg bw/day is used for DNEL derivation until data on the registration substance are available.

Justification for classification or non-classification

No reliable repeated dose toxicity data on the registration substance are available but further testing is ongoing. In the meanwhile, reliable repeated dose toxicity data from a structural analogue was read-across and indicated adverse systemic effects after repeated oral exposure. However, due to deficiencies in the read-across approach the data on the source substance is only used as interim approach for quantitative risk assessment. Therefore, a conclusion on the classification of the registration substance according to Regulation (EC) No. 1272/2008 cannot be made and is thus deemed inconclusive until further repeated dose toxicity data on the registration substance becomes available.