Name: COMPLETE / SUBSTANCE : 1,5,9-Cyclododecatriene / cyclododeca-1,5,9-triene /4904-61-4 Fri, 16 Dec 2022, 15:45:15+0900 /Legal entity owner: National Institute of Health Sciences
Printing date: 2022-12-16T15:45:16.079+09:00

iTable of Contents
0/0 ............................................................................................................................. 1
National Institute of Health Science ..................................................................... 2
1,5,9-Cyclododecatriene ....................................................................................... 3
CORE .......................................................................................................... 3
1 General information .......................................................................... 3
1.10 Assessment approach (assessment entities) ......................... 3
Assessment approach (assessment entities) ........................ 3
OECD .......................................................................................................... 4
D Health Effects .................................................................................. 4
67 Repeated dose toxicity: oral .................................................... 4
Repeated dose toxicity: oral.001 ........................................... 470 Genetic toxicity in vitro ......................................................... 10
Genetic toxicity in vitro.001 ................................................ 10
Genetic toxicity in vitro.002 ................................................ 1573 Toxicity to reproduction ........................................................ 20
Toxicity to reproduction.001 ............................................... 20
DOMAIN .................................................................................................... 26
Substance ......................................................................................... 26
Substance ................................................................................ 26
References ................................................................................................................ 27
Reference Substances ...................................................................................... 27
cyclododeca-1,5,9-triene ............................................................................. 27
Test Materials .................................................................................................. 29
1,5,9-Cyclododecatriene ............................................................................. 29
Literatures ....................................................................................................... 30
Combined Repeated Dose Toxicity Study with theReproduction/
Developmental Toxicity Screening Test on 1,5,9-CYCLODODECATRIENE ....... 30
In Vitro Chromosomal Aberration Test of on 1,5,9-Cyclododecatriene
Cultured Chinese Hamster Cells. ................................................................ 31
Reverse Mutation Test of 1,5,9-Cyclododecatriene on Bacteria. .................... 32
Legal Entities ................................................................................................... 33
National Institute of Health Sciences .......................................................... 33ii

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1,5,9-Cyclododecatriene / cyclododeca-1,5,9-triene / 4904-61-4
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National Institute of Health Science
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LEGAL_ENTITY: National Institute of Health
Science
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General information
Legal entity nameNational Institute of Health Science2

1,5,9-Cyclododecatriene
CORE
General information
Assessment approach (assessment entities)
FIXED_RECORD: Assessment approach

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Remarks:3

OECD
Health Effects
Repeated dose toxicity: oral
ENDPOINT_STUDY_RECORD: Repeated dose toxicity: oral.001

UUID: 087b5c71-30cb-4926-bcde-a32e0e2f24b2
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Administrative data
Endpoint
short-term repeated dose toxicity: oral
Type of information
experimental study
Adequacy of study
key study
Robust study summary
true
Used for classification
false
Used for SDS
false
Reliability
1 (reliable without restriction)
Rationale for reliability incl. deficiencies
guideline study
Reliability 1

Data source
Reference
Combined Repeated Dose Toxicity Study with theReproduction/Developmental Toxicity Screening Teston / MHLW, Japan / study report
Data accessdata published

Materials and methods
4Test guideline
Qualifieraccording to guideline
Guideline
OECD Guideline 422 (Combined Repeated Dose Toxicity Study with the Reproduction /Developmental Toxicity Screening Test)
GLP compliance
yes

Test material
Test material information
1,5,9-Cyclododecatriene
Specific details on test material used for the study
Purity: 94.7% (sum of isomers)

Test animals
Speciesratcommon rodent species
Strainother: Crl:CD(SD)Sexmale/female

Administration / exposure
Route of administration
oral: gavage
Vehiclecorn oilAnalytical verification of doses or concentrations
yes
Duration of treatment / exposure
males: 42 days, females: 41-46 days from 14 days before mating to day 4 of lactation
Frequency of treatment
once a day
Doses / concentrations
Dose / conc.
0mg/kg bw/day (actual dose received)

Dose / conc.
12

mg/kg bw/day (actual dose received)

5

Dose / conc.
60

mg/kg bw/day (actual dose received)

Dose / conc.
300

mg/kg bw/day (actual dose received)

No. of animals per sex per dose
12 animals/sex/doseControl animals
yes, concurrent vehicle
Details on study design- Dose selection rationale:
Dose finding study: Rats were dosed the test substance at 0, 100, 300, and 1000 mg/kg/day for 14
days. Two males died at 1000 mg/kg bw/day. Shiver and reduced body weight gain were observed at 1000 mg/kg bw/day. At 300 mg/kg bw/day and higher, hyperactivity was found. At 100 mg/kg bw/
day and higher, there were slight decreased food consumption and increased liver weight. Based on
the dose finding study, the highest dose for the main test was set as 300 mg/kg bw/day, and middle
and low doses were set as 60 and 12 mg/kg bw/day with a common ratio of 3.
- Post-exposure recovery period in satellite groups: 14 days

Examinations
Observations and examinations performed and frequency
Clinical observation performed and frequency: General condition was observed 3 times a day during
the administration period (before dosing, and immediately after and approximately 2 hours after
dosing) and once a day (in the morning) during the recovery period.
Body weights were determined on days 1, 4, 8, 11, 15, 18, 22, 25, 29, 32, 36, 39 and 42 of adminis
tration for males and on days 1, 4, 8, 11 and 15 of administration, days 0, 4, 7, 11, 14, 17 and 20 of
gestation and days 0 and 4 of lactation for females, and the day of necropsy in males and females. In addition, males and females in the recovery groups were weighed on days 1, 4, 8, 11 and 14 ofrecovery and on the day of necropsy.
Food consumption was determined on days 1, 4, 8, 11, 15, 32, 36, 39 and 42 of administration for males and on days 1, 4, 8, 11 and 15 of administration, days 1, 4, 7, 11, 14, 17 and 20 of gestation and days
2 and 4 of lactation in females, but it was not determined during the mating period for males andfemales. In addition, it was determined on days 1, 4, 8, 11 and 14 of recovery for males and females
in the recovery groups.
Sacrifice and pathology
Necropsy: Detailed macroscopic examination was then conducted on the organs/tissues throughout
the body of each animal, including the external appearance, head, thorax and abdomen.
Measurement of organ weighs: The brain, thyroids (including parathyroids), adrenals, thymus, spleen,
heart, liver, kidneys, testes, epididymides were determined.
Histopathological examination: The liver in both sexes and the kidney in males of all groups, thecerebrum, cerebellum, pituitary, spinal cord (thoracic), sciatic nerve, thyroids, parathyroids, adrenals,
thymus, spleen, submandibular lymph node, mesenteric lymph node, heart, lung (including bronchus),
duodenum, jejunum, ileum, cecum, colon, rectum, liver, kidneys, urinary bladder, testes, epididymides,
ovaries, uterus, seminal vesicles, sternum (including bone marrow), femur (including bone marrow)6

in males and females at 0 and 300 mg/kg. In addition, all gross pathological lesions of all animals
were examined.
Statistics
Statistical methods: Dunnett's test for continuous data and Dunnett-type mean rank test for quantal
data were used.

Results and discussion
Results of examinations
Clinical signs
effects observed, treatment-related
Description (incidence and severity)
Salivation caused by irritation from the test article was observed during the administration period inboth sexes in the 300 mg/kg group. Salivation was observed sporadically at approximately 2 hours
after administration from week 3 of administration.
Mortality
no mortality observed
Body weight and weight changes
effects observed, treatment-related
Description (incidence and severity)
Reduced body weight gain was observed in males during the administration period and in females
from day 14 of gestation in the 300 mg/kg group.
Food consumption and compound intake (if feeding study)no effects observed
Haematological findings
effects observed, treatment-related
Description (incidence and severity)
At the end of administration period, prolongation of prothrombin time in males in the 60 mg/kg group, and prolongation of prothrombin time and activated partial thromboplastin time and high value in f
ibrinogen in males in the 300 mg/kg group were observed. Additionally, a low value in white blood cellcount with low values in lymphocyte and neutrophil counts and basophil percentage and count wasobserved in females in the 300 mg/kg group.
Clinical biochemistry findings
effects observed, treatment-related
Description (incidence and severity)
γ-GTP, total protein, and albumin were increased at 300 mg/kg bw/day in males. AST, ALT, and creatin
ine were decreased at 300 mg/kg bw/day in females.
Urinalysis findings
no effects observed
Behaviour (functional findings)
no effects observed
Description (incidence and severity)
Manipulative test, measurements of grip strength and motor activity: No test article-related changes
were observed in males or females.
7Organ weight findings including organ / body weight ratios
effects observed, treatment-related
Description (incidence and severity)
At the end of the administration period, high value in the liver weight in both sexes in the 300 mg/kg
group was observed. High value in the liver weight without hypertrophy of centrilobular hepatocytes
in females in the 60 mg/kg group was observed. Additionally, high value in the kidney weight wasobserved in males in the 300 mg/kg group.
Histopathological findings: non-neoplastic
effects observed, treatment-related
Description (incidence and severity)
At the end of the administration period, histopathology revealed hypertrophy of centrilobular hepa
tocytes in both sexes.
There were appearance of eosinophilic bodies in tubular epithelium caused by α2µ-globulin in 1, 2 and5 males in the 12, 60 and 300 mg/kg groups, respectively, and the incidence was increased in the 300
mg/kg group.

Effect levels
Key result
false
Dose descriptor
NOAEL
Effect level12

mg/kg bw/day (actual dose received)

Based onact. ingr.
Sexmale/female
Basis for effect level
haematology
prolonged PT in males at 60 mg/kg bw/day
organ weights and organ / body weight ratios
increased liver weight in females at 60 mg/kg bw/day

Any other information on results incl. tables
Figures and Tables (in English) are available in the following full report of the study.
http://dra4.nihs.go.jp/mhlw_data/home/pdf/PDF4904-61-4b.pdf

Applicant's summary and conclusion
Conclusions
Based on the effects on the liver of cyclododeca-1, 5, 9-trieneat 60 mg/kg bw/day, the NOAEL for its
repeated-dose toxicity was determined to be 12 mg/kg bw/day in rats.
8

Executive summary

A combined repeated oral dose toxicity study with the reproduction/developmental toxicity screening
test was performed in accordance with OECD TG 422. Male and female rats (12 animals/sex/dose) wereadministered 1,5,9-cyclododecatriene at 0 (vehicle: corn oil), 12, 60, and 300 mg/kg bw/day. Males were
dosed for 42 days, including a 14-day pre-mating period and subsequent mating period. Females were
dosed for 42–53 days, including 14-day pre-mating, mating, and gestation periods, and until lactation
day 4. Five out of 12 males dosed at 0 and 300 mg/kg bw/day were treated as a recovery group. Reduced
body weight gain was observed in both sexes in the 300 mg/kg bw/day group. Regarding hematologicalparameters, prolongation of prothrombin time in males in the 60 and 300 mg/kg bw/day groups as wellas activated partial thromboplastin time and a high level of fibrinogen in males in the 300 mg/kg bw/day
group were observed. High liver weights were also observed in both sexes at 300 mg/kg bw/day and infemales at 60 mg/kg bw/day. Moreover, histopathological analysis revealed hypertrophy of centrilobular
hepatocytes at 300 mg/kg bw/day. These changes were either not found or the degree and incidence
were reduced after the recovery period. Based on the effects on the liver of 1,5,9-cyclododecatriene 60
mg/kg bw/day, the NOAEL for its repeated-dose toxicity was determined to be 12 mg/kg bw/day in rats.

9

Genetic toxicity in vitro
ENDPOINT_STUDY_RECORD: Genetic toxicity in vitro.001

UUID: e351c082-0693-421e-aab6-f843c868f26c
Dossier UUID:
Author:Date: 2022-12-16T15:43:21.627+09:00
Remarks:

Administrative data
Endpoint
in vitro gene mutation study in bacteria
Type of information
experimental study
Adequacy of study
key study
Robust study summary
false
Used for classification
false
Used for SDS
false
Reliability
1 (reliable without restriction)
Rationale for reliability incl. deficiencies
guideline study
Reliability 1

Data source
Reference
Reverse Mutation Test of 1,5,9-Cyclododecatriene on Bacteria. / MHLW, Japan / study report
Data accessdata published

Materials and methods
Test guideline
Qualifieraccording to guideline
Guideline
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
in vitro gene mutation study in bacteria
GLP complianceyes
10Type of assay
bacterial reverse mutation assay
in vitro gene mutation study in bacteria

Test material
Test material information
1,5,9-Cyclododecatriene
Specific details on test material used for the study
Purity 96.6%

MethodSpecies / strain
Species / strain / cell typeS. typhimurium TA 1535, TA 1537, TA 98, TA 100 and E. coli WP2bacteria
Metabolic activation
with and without
Metabolic activation system
S9 mix: Rat liver, induced with phenobarbital and 5,6-benzoflavone
Test concentrations with justification for top dose
-S9 mix: 0, 0.15, 0.31, 0.61, 1.22, 2.44, 4.88 μg/plate(TA100, TA1535,
TA1537)0, 0.61, 1.22, 2.44, 4.88, 9.77, 19.5 μg/plate(TA98, WP2 uvrA)+S9 mix: 0, 2.44, 4.88, 9.77, 19.5, 39.1, 78.1 μg/plate(TA strains)
0, 9.77, 19.5, 39.1, 78.1, 156, 313 μg/plate(WP2 uvrA)
Maximum concentration was established based on the result of the preliminary test at concentration
up to 5000 ug/plate. In this test, the growth inhibition was observed at 4.88 μg/plate and above forS. typhimurium TA100, 1535 and 1537 without S9 mix, at 19.5 μg/plate and above for S. typhimurium
TA98 and E. coli WP2 uvrA without S9 mix, at 78.1 μg/plate and above for S. typhimurium TA strains
with S9 mix and at 313 μg/plate and above for E. coli WP2 uvrA with S9 mix.
Vehicle / solvent
AcetoneControls
Negative solvent / vehicle controls
yes
Positive controls
yes
Positive control substancesodium azide
without S9 mix (TA 1535)
benzo(a)pyrene
with S9 mix (TA100, TA98, TA1537)
other: without S9 mix:2-(2-Furyl)-3-(5-nitro -2-furyl)acrylamide (TA100, TA98, WP2uvrA), with S9 mix:
2-Aminoanthracene (TA1535, WP2 uvrA)
Details on test system and experimental conditions
METHOD OF APPLICATION: Preincubation
11

DURATION- Preincubation period: 20 min at 37°C
- Exposure duration:48 hrs
NUMBER OF PLATES: 3
NUMBER OF REPLICATIONS: 2
DETERMINATION OF CYTOTOXICITY- Method: other: growth inhibition
Evaluation criteria
A chemical was judged to be mutagenic when the mean number of revertant colonies per plateincreased more than twice that of the negative control and when the dose-related and reproducible
increase was observed.
Statistics
not used

Results and discussion
Test results
Key result
false
Species / strain
S. typhimurium TA 100
bacteria
Metabolic activation
with and without
Genotoxicity
negative
Cytotoxicity / choice of top concentrations
cytotoxicity -S9 mix: 4.88 μg/plate, +S9 mix: 78.1 μg/plate
Vehicle controls validity
valid
Positive controls validity
valid
Key result
false
Species / strain
S. typhimurium TA 1535
bacteria
Metabolic activation
with and without
Genotoxicity
negative
Cytotoxicity / choice of top concentrations
cytotoxicity -S9 mix: 4.88 μg/plate, +S9 mix: 39.1 μg/plate
Key result
false
Species / strain
E. coli WP2 uvr A pKM 101
12

bacteria
Metabolic activation
with and without
Genotoxicity
negative
Cytotoxicity / choice of top concentrations
cytotoxicity -S9 mix: 9.77 μg/plate, +S9 mix: 156 μg/plate
Vehicle controls validity
valid
Positive controls validity
valid
Key result
false
Species / strain
S. typhimurium TA 98
bacteria
Metabolic activation
with and without
Genotoxicity
negative
Cytotoxicity / choice of top concentrations
cytotoxicity -S9mix: 9.77 μg/plate, +S9 mix: 156 μg/plate
Vehicle controls validity
valid
Positive controls validity
valid
Key result
false
Species / strain
S. typhimurium TA 1537
bacteria
Metabolic activation
with and without
Genotoxicity
negative
Cytotoxicity / choice of top concentrations
cytotoxicity -S9 mix: 4.88 μg/plate, +S9 mix: 39.1 μg/plate
Vehicle controls validity
valid
Positive controls validity
valid
Additional information on results
There were no precipitation in any test concentration.
13

Cytotoxic concentration: Growth inhibition was observed at 4.88 μg/plate and above for S. typh
imurium TA100, TA1535 and TA1537 without S9 mix, at 9.77 μg/plate and above for S. typhimurium
TA98 and E. coli WP2 uvrA without S9 mix, at 39.1 μg/plate and above for S. typhimurium TA1535
and TA1537 with S9 mix, at 78.1μg/plate and above for S. typhimurium TA100 and TA98 with S9 mix
and at 156 μg/plate and above for E. coli WP2 uvrA with S9 mix.
Genotoxic effects:

Any other information on results incl. tables

Figures and Tables (in Japanease) are available in the following full report of the study.
http://dra4.nihs.go.jp/mhlw_data/home/pdf/PDF4904-61-4e.pdf
Tables (in English) are attachted to this document. Please download the export file to see the Tables.

Overall remarks, attachments
Attachments
Attached (sanitised) documents for publication
4904-61-4_Ames Tables.xlsx / 34.59 KB (application/vnd.openxmlformatsofficedocument.spreadsheetml.sheet)

Applicant's summary and conclusion
Conclusions
Genotoxic effects:
With metabolic activation: Negative
Without metabolic activation: Negative
Executive summary
In the bacterial reverse mutation assay using S. typhimurium TA100, TA1535, TA98, and TA1537, andE. coli WP2uvrA/pKM101 (OECD TG 471), negative results were obtained for 1,5,9-cyclododecatrienewith or without metabolic activation.

14

ENDPOINT_STUDY_RECORD: Genetic toxicity in vitro.002

UUID: 86d5e6eb-2f49-4f10-a61d-c382dcf83394
Dossier UUID:
Author:Date: 2022-12-16T15:44:03.408+09:00
Remarks:

Administrative data
Endpoint
in vitro cytogenicity / chromosome aberration study in mammalian cells
Type of information
experimental study
Adequacy of study
key study
Robust study summary
true
Used for classification
false
Used for SDS
false
Reliability
1 (reliable without restriction)
Rationale for reliability incl. deficiencies
guideline study
Reliability 1

Data source
Reference
In Vitro Chromosomal Aberration Test of on 1,5,9-Cyclododecatriene Cultured Chinese Hamster Cells. /MHLW, Japan / study report
Data accessdata published

Materials and methods
Test guideline
Qualifieraccording to guideline
Guideline
OECD Guideline 473 (In Vitro Mammalian Chromosome Aberration Test)in vitro cytogenicity / chromosome aberration study in mammalian cells
GLP complianceyes
15Type of assay
in vitro mammalian chromosome aberration testin vitro cytogenicity / chromosome aberration study in mammalian cells

Test material
Test material information
1,5,9-Cyclododecatriene
Specific details on test material used for the study
Purity 96.6%

MethodSpecies / strain
Species / strain / cell typeother: Chinese hamster lung(CHL/IU) cell
Metabolic activation
with and without
Metabolic activation system
S9 mix: Rat liver, induced with phenobarbital and 5,6- benzoflavone
Test concentrations with justification for top dose
-S9 mix(short-term treatment): 0, 44.0, 57.2, 74.4, 96.7 μg/mL+S9 mix(short-term treatment): 0, 13.8, 19.4, 27.1, 37.9, 53.1 μg/mL
Preliminary study
0, 13.3, 26.6, 53.1, 106, 213, 425, 850, 1700 μg/mL (=10 mM)
50% growth inhibition was observed
at 53.1 μg/mL with S9 mix
at 106 μg/mL without S9 mix
at 65.8 μg/mL 24h
at 63.5 μg/mL 48 h

Vehicle / solvent
AcetoneControls
Untreated negative controls
yes
Negative solvent / vehicle controls
yes
Positive controls
yes
Positive control substancecyclophosphamide+S9 mix
16

mitomycin C- S9 mix
Details on test system and experimental conditions
METHOD OF APPLICATION: Exposure duration: [short-term treatment]:6 hrs + 18 hr,
STAIN: Giemsa stain (3 v/v%) for 15 min.
NUMBER OF REPLICATIONS: 2
NUMBER OF CELLS EVALUATED: 100 + 100 cells /concentration
DETERMINATION OF CYTOTOXICITY- Method: relative total growth
Evaluation criteria
For the evaluation of the frequencies of structural aberrations and of polyploidy induced, the following
criteria were employed. For the evaluation of the frequencies of structural aberrations and of polyploi
dy induced, the following criteria were employed. Appearance incidence of cell with chromosomal abe
rrations: Negative(-): less than 5%, Equivocal(±): 5% or more and less than 10%, Positive(+): 10% or
more
Statistics
Not used

Results and discussion
Test results
Key result
false
Species / strainother: Chinese hamster lung(CHL/IU) cells
Metabolic activation
with
Genotoxicity
positive clastogenicity
Cytotoxicity / choice of top concentrations
cytotoxicity
Vehicle controls validity
valid
Untreated negative controls validity
valid
Positive controls validity
valid
Key result
false
Species / strainother: Chinese hamster lung(CHL/IU) cells
Metabolic activation
with
Genotoxicity
ambiguous polyploidy
17

Cytotoxicity / choice of top concentrations
cytotoxicity
Vehicle controls validity
valid
Untreated negative controls validity
valid
Positive controls validity
valid
Key result
false
Species / strainother: Chinese hamster lung(CHL/IU) cells
Metabolic activation
without
Genotoxicity
negative
Cytotoxicity / choice of top concentrations
cytotoxicity
Vehicle controls validity
valid
Untreated negative controls validity
valid
Positive controls validity
valid
Additional information on results
Increase in structural and numerical chromosomal aberrations was observed in the short-term
treatment with metabolic activation, while not observed in the short-term treatment without metabo
lic activation.

Any other information on results incl. tables
Figures and Tables (in English) are available in the following full report of the study.
http://dra4.nihs.go.jp/mhlw_data/home/pdf/PDF4904-61-4f.pdf

Applicant's summary and conclusion
Conclusions
Positive with S9 mix
Executive summary
In an in vitro chromosomal aberration test using CHL/IU cells (OECD TG 473), 1,5,9cyclododecatriene was clastogenic with metabolic activation and had weak potential to induce18

polyploidy with metabolic activation. Based on these results, 1,5,9-cyclododecatriene was judged tocause chromosomal aberration in vitro.

19

Toxicity to reproduction
ENDPOINT_STUDY_RECORD: Toxicity to reproduction.001

UUID: bc99dfd6-e5f8-4f59-abe7-333bd20e3176
Dossier UUID:
Author:Date: 2022-12-16T15:44:47.259+09:00
Remarks:

Administrative data
Endpoint
screening for reproductive / developmental toxicity
Type of information
experimental study
Adequacy of study
key study
Robust study summary
true
Used for classification
false
Used for SDS
false
Reliability
1 (reliable without restriction)
Rationale for reliability incl. deficiencies
guideline study
Reliability 1

Data source
Reference
Reverse Mutation Test of 1,5,9-Cyclododecatriene on Bacteria. / MHLW, Japan / study report
Data accessdata published

Materials and methods
Test guideline
Qualifieraccording to guideline
Guideline
OECD Guideline 422 (Combined Repeated Dose Toxicity Study with the Reproduction /Developmental Toxicity Screening Test)
GLP complianceyes
20Test material
Test material information
1,5,9-Cyclododecatriene
Specific details on test material used for the study
Purity: 94.7% (sum of isomers)

Test animals
Speciesrat
Strainother: Crl:CD(SD)Sexmale/female

Administration / exposure
Route of administration
oral: gavage
Vehiclecorn oil
Details on mating procedure
Males and females in the same dose group of the main groups were co-housed overnight on a oneto-one basis after the end of the pre-mating administration period. Copulation was considered s
uccessful if the formation of vaginal plugs or presence of sperm in vaginal smears was confirmed
the following morning. The length of the mating period for the same male and female was 4 days at
maximum.
Delivery and delivery/lactation status: All copulated females were allowed to deliver spontaneously
and examined for any abnormality of delivery. Dams which completed delivery were observed forclearance of placenta and amnion, and the end of delivery was designated as day 0 of lactation. D
ams were then allowed to nurse their liveborn pups until day 4 of lactation and examined for lactation
status using the gathering of pups, nesting and lactating as indicators
Analytical verification of doses or concentrations
yes
Duration of treatment / exposure
males: 42 days, females: 41-46 days from 14 days before mating to day 4 of lactation
Frequency of treatment
once a day
Doses / concentrations
Dose / conc.
0mg/kg bw/day (actual dose received)

Dose / conc.
12

mg/kg bw/day (actual dose received)

21

Dose / conc.
60

mg/kg bw/day (actual dose received)

Dose / conc.
300

mg/kg bw/day (actual dose received)

No. of animals per sex per dose
12 animals/sex/doseControl animals
yes, concurrent vehicle

Examinations
Parental animals: Observations and examinations
Clinical observation performed and frequency: General condition was observed 3 times a day during
the administration period (before dosing, and immediately after and approximately 2 hours after
dosing) and once a day (in the morning) during the recovery period.
Body weights were determined on days 1, 4, 8, 11, 15, 18, 22, 25, 29, 32, 36, 39 and 42 of adminis
tration for males and on days 1, 4, 8, 11 and 15 of administration, days 0, 4, 7, 11, 14, 17 and 20 of
gestation and days 0 and 4 of lactation for females, and the day of necropsy in males and females. In addition, males and females in the recovery groups were weighed on days 1, 4, 8, 11 and 14 ofrecovery and on the day of necropsy.
Food consumption was determined on days 1, 4, 8, 11, 15, 32, 36, 39 and 42 of administration for males and on days 1, 4, 8, 11 and 15 of administration, days 1, 4, 7, 11, 14, 17 and 20 of gestation and days
2 and 4 of lactation in females, but it was not determined during the mating period for males andfemales. In addition, it was determined on days 1, 4, 8, 11 and 14 of recovery for males and females
in the recovery groups.
Oestrous cyclicity (parental animals)
Vaginal smears were collected from all females in the main groups and microscopically examined
every day (in the morning) from the day after the start of administration until the day copulation was
confirmed. Vaginal smear pictures were classified as proestrus, estrus, metestrus and diestrus andexamined for the frequency of estrus and interval between estruses (estrous cycle).
Postmortem examinations (offspring)
The numbers of liveborn pups and stillborn pups were counted on the day of birth. After liveborn pu
ps were examined for any external abnormality, sexed and weighed, dams were allowed to nurse theirpups. Liveborn pups were observed for mortality once daily until day 4 after birth. All liveborn pups
were exsanguinated after measurement of body weight on day 4 after birth, necropsied and examined
for any abnormality in organs/tissues, including those in the head, thorax and abdomen. Individual
body weights of liveborn pups were recorded, and the average body weight per litter was calculated
by sex.
Pathological examinations were performed.
Statistics
Dunnett's test for continuous data, Dunnett-type mean rank test for quantal data and chi-square testwith Yates’ continuity correction or chi-square test with Yates’ continuity correction for other data
were used.
Reproductive indicesNo. of copulated animals, No. of males that impregnated females, No. of pregnant females, No. offemales that delivered liveborn pups, estrous cycle, gestational length, No.of corpora lutea, No. of im
plantation sites, total No. of liveborn and stillborn pups, No. of liveborn pups, sex ratio on day 0 and
22

day 4 after birth, copulation index (No. of copulated animals / No. of animals housed together x 100),
insemination index (No. of pregnant females / No. of copulated males x 100), fertility index (No. of
pregnant females / No. of copulated females x 100), delivery index (No. of females that delivered liv
eborn pups / No. of pregnant females x 100), implantation index (No. of implantation sites / No. of
corpora lutea x100), stillbirth index (No. of stillborn pups / No. of pups born x 100)
Offspring viability indices
Index of external abnormalities (No. of pups with external abnormalities / No. of pups born x 100), live
birth index (No. of liveborn pups / No. of pups born x 100), and viability index on day 4 after birth (No.
of live pups on day 4 after birth / No. of liveborn pups x 100) were determined.

Results and discussion
Results: P0 (first parental generation)
General toxicity (P0)
Clinical signs
effects observed, treatment-related
Description (incidence and severity)
Salivation caused by irritation from the test article was observed during the administration period inboth sexes in the 300 mg/kg group. Salivation was observed sporadically at approximately 2 hours
after administration from week 3 of administration.
Mortality
no mortality observed
Body weight and weight changes
effects observed, treatment-related
Description (incidence and severity)
Reduced body weight gain was observed in males during the administration period and in females
from day 14 of gestation in the 300 mg/kg group.
Food consumption and compound intake (if feeding study)no effects observed
Haematological findings
effects observed, treatment-related
Description (incidence and severity)
At the end of administration period, prolongation of prothrombin time in males in the 60 mg/kg group, and prolongation of prothrombin time and activated partial thromboplastin time and high value infibrinogen in males in the 300 mg/kg group were observed. Additionally, a low value in white blood
cell count with low values in lymphocyte and neutrophil counts and basophil percentage and count w
as observed in females in the 300 mg/kg group.
Clinical biochemistry findings
effects observed, treatment-related
Description (incidence and severity)
γ-GTP, total protein, and albumin were increased at 300 mg/kg bw/day in males. AST, ALT, andcreatinine were decreased at 300 mg/kg bw/day in females.
Urinalysis findings
no effects observed
Behaviour (functional findings)
no effects observed
23Organ weight findings including organ / body weight ratios
effects observed, treatment-related
Description (incidence and severity)
At the end of the administration period, high value in the liver weight in both sexes in the 300 mg/kg
group was observed. High value in the liver weight without hypertrophy of centrilobular hepatocytes
in females in the 60 mg/kg group was observed. Additionally, high value in the kidney weight wasobserved in males in the 300 mg/kg group.
Histopathological findings: non-neoplastic
effects observed, treatment-related
Description (incidence and severity)
At the end of the administration period, histopathology revealed hypertrophy of centrilobular hepa
tocytes in both sexes.
There were appearance of eosinophilic bodies in tubular epithelium caused by α2μ-globulin in 1, 2
and 5 males in the 12, 60 and 300 mg/kg groups, respectively, and the incidence was increased inthe 300 mg/kg group.

Reproductive function / performance (P0)
Reproductive function: oestrous cycle
no effects observed
Reproductive performance
effects observed, treatment-related
Description (incidence and severity)
A low value in the number of liveborn pups was observed in the 300 mg/kg group. There were no test
article-related effects on the estrous cycle, number of elapsed days until copulation, copulation index,
insemination index or fertility index. In addition, no test article-related effects were observed in thedelivery index, duration of gestation, numbers of corpora lutea and implantation sites, implantation index, stillborn index, birth index or sex ratio. No abnormalities were found in the lactation statusduring the lactation period.

Effect levels (P0)
Key result
false
Dose descriptor
NOAEL
Effect level60

mg/kg bw/day (actual dose received)

Based onact. ingr.
Sexmale/female
Basis for effect level
other: a significantly low value of the number of liveborn pups at 300 mg/kg bw/day

Results: F1 generation
24

General toxicity (F1)Mortality / viability
mortality observed, treatment-related
Description (incidence and severity)
A significantly low value of the number of liveborn pups was observed in the 300 mg/kg bw/day
group.
Body weight and weight changes
no effects observed

Any other information on results incl. tables
Figures and Tables (in English) are available in the following full report of the study.
http://dra4.nihs.go.jp/mhlw_data/home/pdf/PDF4904-61-4b.pdf

Applicant's summary and conclusion
Conclusions
A significantly low value of the number of liveborn pups was observed in the 300 mg/kg bw/day
group. No other effects were observed for fertility and development. The NOAEL for the rat repro
ductive/developmental toxicity of cyclododeca-1, 5, 9-triene was determined to be 60 mg/kg bw/day,at which maternal general toxicity was observed as described above.
Executive summary
A combined repeated oral dose toxicity study with the reproduction/developmental toxicity screening
test was performed in accordance with OECD TG 422. Male and female rats (12 animals/sex/dose) wereadministered 1,5,9-cyclododecatriene at 0 (vehicle: corn oil), 12, 60, and 300 mg/kg bw/day. Males were
dosed for 42 days, including a 14-day pre-mating period and subsequent mating period. Females were
dosed for 42–53 days, including 14-day pre-mating, mating, and gestation periods, and until lactation
day 4. Five out of 12 males dosed at 0 and 300 mg/kg bw/day were treated as a recovery group. Reduced
body weight gain was observed in both sexes in the 300 mg/kg bw/day group. Regarding hematologicalparameters, prolongation of prothrombin time in males in the 60 and 300 mg/kg bw/day groups as wellas activated partial thromboplastin time and a high level of fibrinogen in males in the 300 mg/kg bw/day
group were observed. High liver weights were also observed in both sexes at 300 mg/kg bw/day and infemales at 60 mg/kg bw/day. Moreover, histopathological analysis revealed hypertrophy of centrilobular
hepatocytes at 300 mg/kg bw/day. These changes were either not found or the degree and incidence
were reduced after the recovery period. A significantly low value of the number of liveborn pups wasobserved in the 300 mg/kg bw/day group. No other effects were observed for fertility and development.
The NOAEL for the rat reproductive/developmental toxicity of 1,5,9-cyclododecatriene was determined
to be 60 mg/kg bw/day, at which maternal general toxicity was observed as described above.

25

DOMAIN
Substance
SUBSTANCE: 1,5,9-Cyclododecatriene
UUID: e16ca988-a4be-4cd9-9dc8-64a4e1e8f212
Dossier UUID:
Author:Date: 2022-12-16T15:45:04.247+09:00
Remarks:
Substance name
1,5,9-Cyclododecatriene
Legal entity
National Institute of Health Sciences / Kawasaki / Japan

Identification of substance
Reference substancecyclododeca-1,5,9-triene / cyclododeca-1,5,9-triene / 4904-61-4 / 225-533-8
EC number

EC name225-533-8

EC Inventory

CAS number

CAS name

4904-61-4
IUPAC namecyclododeca-1,5,9-triene

Role in the supply chain
Manufacturer
false
Importer
false
Only representativefalse
Downstream user
false

26

References
Reference Substances
REFERENCE_SUBSTANCE: cyclododeca-1,5,9-triene
UUID: ECB5-32e2ca12-bb3c-4055-929c-602a4bce5dc7
Dossier UUID:
Author:Date: 2018-03-08T10:45:13.000+09:00
Remarks:
Reference substance namecyclododeca-1,5,9-triene
IUPAC namecyclododeca-1,5,9-triene

Inventory
Inventory number
Inventory namecyclododeca-1,5,9-triene
Inventory
EC Inventory
Inventory number
225-533-8
CAS number
4904-61-4
Molecular formulaC12H18Description
CAS number
4904-61-4

Synonyms
Synonyms
Identity
1,5,9-Cyclododecatriene

Molecular and structural information
Molecular formulaC12H1827

Molecular weight
162.2713
SMILES notation
C1C\C=C\CC\C=C\CC\C=C/1
InChl
InChI=1/C12H18/c1-2-4-6-8-10-12-11-9-7-5-3-1/h1-2,7-10H,3-6,11-12H2
Structural formula28Test Materials
TEST_MATERIAL_INFORMATION: 1,5,9Cyclododecatriene
UUID: a87598f4-1681-4f77-bb10-1b3effa8b6cd
Dossier UUID:
Author:Date: 2018-03-08T10:47:51.000+09:00
Remarks:
Name
1,5,9-Cyclododecatriene

29Literatures
LITERATURE: Combined Repeated Dose Toxicity Studywith theReproduction/Developmental Toxicity Screening
Test on 1,5,9-CYCLODODECATRIENEUUID: fa643fc0-02bc-4b23-a9b3-38dce416c64b
Dossier UUID:
Author:Date: 2018-03-08T16:53:19.000+09:00
Remarks:

General information
Reference Typestudy report
Title
Combined Repeated Dose Toxicity Study with theReproduction/Developmental Toxicity Screening
Test on 1,5,9-CYCLODODECATRIENE
Author
MHLW, Japan
Year
2008
Bibliographic source
Japan Existing Chemical Data Base (JECDB) http://dra4.nihs.go.jp/mhlw_data/jsp/Se archPage
ENG.jsp
Testing facility
Bozo Research Center Inc.

30

LITERATURE: In Vitro Chromosomal Aberration Test of on
1,5,9-Cyclododecatriene Cultured Chinese Hamster Cells.
UUID: 190607e5-f62e-4c0d-8caa-6826131d2902
Dossier UUID:
Author:Date: 2018-03-08T15:29:36.000+09:00
Remarks:

General information
Reference Typestudy report
Title
In Vitro Chromosomal Aberration Test of on 1,5,9-Cyclododecatriene Cultured Chinese Hamster Cells.
Author
MHLW, Japan
Year
2008
Bibliographic source
Japan Existing Chemical Data Base (JECDB) http://dra4.nihs.go.jp/mhlw_data/jsp/Sea rchPage
ENG.jsp
Testing facility
Bozo Research Center Inc.

31

LITERATURE: Reverse Mutation Test of 1,5,9Cyclododecatriene on Bacteria.
UUID: 6470f598-9801-49b9-8c09-d092c960b48f
Dossier UUID:
Author:Date: 2018-03-08T10:25:04.000+09:00
Remarks:

General information
Reference Typestudy report
Title
Reverse Mutation Test of 1,5,9-Cyclododecatriene on Bacteria.
Author
MHLW, Japan
Year
2008
Bibliographic source
Bibliographic source JECDB http://dra4.nihs.go.jp/mhlw_data/jsp/SearchPageENG.jsp
Testing facility
Bozo Research Center Inc.

32

Legal Entities
LEGAL_ENTITY: National Institute of Health Sciences
UUID: IUC4-b036ff75-0f3c-323b-b200-ed5f46cf5101
Dossier UUID:
Author:Date: 2022-11-07T15:49:29.000+09:00
Remarks:

General information
Legal entity nameNational Institute of Health Sciences
Remarks
Disclaimer: The contents in this document were created based on the MHLW (Ministry of Health,Labour and Welfare) peer reviewed study reports (in Japanese) in JECDB (Japan Existing Chemical
Database) at http://dra4.nihs.go.jp/mhlw_data/jsp/SearchPageENG.jsp. Authorship is in the Divisionof Risk Assessment, the National Institute of Health Sciences, and the contents do not reflect any official MHLW opinions or any other regulatory policies.

Address
Address 1
Tonomachi 3-25-26
Address 2
Kawasaki-ku
Postal code
210-9501
Town
KawasakiRegion / State
Kanagawa
CountryJapan
JP

Identifiers
Other IT system identifiers
IT system
LEO
ID
10767
IT systemIUCLID433

ID
16558402024DIV750

34