Registration Dossier

Administrative data

Endpoint:
developmental toxicity
Type of information:
experimental study
Adequacy of study:
key study
Study period:
22 November 2016 - 28 November 2017
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Cross-reference
Reason / purpose:
reference to other study
Reference
Endpoint:
repeated dose toxicity: oral, other
Remarks:
dose range finding study
Type of information:
experimental study
Adequacy of study:
supporting study
Study period:
28 July 2016 - 16 January 2017
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
test procedure in accordance with generally accepted scientific standards and described in sufficient detail
Qualifier:
no guideline available
Principles of method if other than guideline:
- Principle of test: The toxicity of CAPA 2043, after daily oral administration (gavage) to rats, was investigated over a period of 2 consecutive weeks.
- Short description of test conditions: Three groups, each of 4 male and 4 female Sprague Dawley SD rats, received the test item at dose levels of 100, 300 and 1000 mg/kg/day for 2 consecutive weeks. A fourth similarly constituted group received the vehicle alone (corn oil) and acted as a control.
- Parameters analysed / observed: clinical signs, body weight and food consumption, haematology and clinical chemistry, post mortem organ weights and macroscopic observations.
GLP compliance:
yes
Limit test:
no
Specific details on test material used for the study:
Identity: CAPA 2043
Label name: CAPA(TM) 2043
Chemical name: 2-oxepanone, polymer with 1,4-butanediol
Batch number: WAC000478
CAS number: 31831-53-5
EC number: 608-670-1
Expiry date: 10 May 2018
Appearance: Clear, colourless liquid
Storage conditions: Cool, dry conditions in well-scaled receptacles
Species:
rat
Strain:
Sprague-Dawley
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Envigo RMS s.r.l., San Pietro al Natisone (UD), Italy
- Age at arrival: 27-29 days old
- Weight at arrival: 75-99 g
- Housing: up to 5 of one sex in a cage
- Diet (e.g. ad libitum): Laboratory rodent diet (4 RF 21, Mucedola S.r.l., Via G. Galilei, 4, 20019 Settimo Milanese (MI), Italy); ad libitum
- Water (e.g. ad libitum): Drinking water; ad libitum
- Acclimation period: 12 days


ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 2 °C
- Humidity (%): 55 ± 15%
- Air changes (per hr): 15 - 20
- Photoperiod (hrs dark / hrs light): 12 hrs dark/12 hrs light
Route of administration:
oral: gavage
Details on route of administration:
The test item was administered orally, by gavage at a dose volume of 5mL/kg. Control animals received the vehicle alone at the same dose volume. The dose was administered to each animal on the basis of the most recently recorded body weight and the volume administered was recorded for each animal.
Vehicle:
corn oil
Details on oral exposure:
- PREPARATION OF DOSING SOLUTIONS: The required amount of CAPA 2043 was suspended in the vehicle. The formulations were prepared daily (concentrations of 20, 60 and 200 mg/mL). Concentrations were calculated and expressed in terms of test item as supplied. The formulations were prepared daily (concentrations of 20, 60 and 200 mg/mL). Concentrations were calculated and expressed in terms of test item as supplied.

- VEHICLE: corn oil
Analytical verification of doses or concentrations:
no
Details on analytical verification of doses or concentrations:
n.a.
Duration of treatment / exposure:
2 weeks
Frequency of treatment:
Once a day, 7 days a week
Dose / conc.:
0 mg/kg bw/day (nominal)
Remarks:
Vehicle (corn oil) control
Dose / conc.:
100 mg/kg bw/day (nominal)
Remarks:
Low dose group
Dose / conc.:
300 mg/kg bw/day (nominal)
Remarks:
Mid dose group
Dose / conc.:
1 000 mg/kg bw/day (nominal)
Remarks:
High dose group
No. of animals per sex per dose:
4
Control animals:
yes, concurrent vehicle
Details on study design:
Three groups, each of 4 male and 4 female Sprague Dawley SD rats, received the test item at dose levels of 100, 300 and 1000 mg/kg/day for 2 consecutive weeks. A fourth similarly constituted group received the vehicle alone (corn oil) and acted as a control.
-Dose selection rationale: not reported
- Rationale for animal assignment: Each group comprised 4 male and 4 female rats. The group identification and animal numbers assigned to the treatment are summarized below:
Group 1 (0 mg/kg bw/day) Rat numbers: males (even) 2-8, females (odd) 1-7
Group 2 (100 mg/kg bw/day) Rat numbers: males (even) 10-16, females (odd) 9-15
Group 3 (300 mg/kg bw/day) Rat numbers: males (even) 18-24, females (odd) 17-23
Group 4 (1000 mg/kg bw/day) Rat numbers: males (even) 26-32, females (odd) 25-31
The rat numbers listed above formed the last digits of a computer generated 8 figure animal number (the remaining digits of the animal number were different for each concurrent study and served to ensure unique animal numbering for any study employing computerised data collection). The computerised system used in this study was Pristima, Version 6.4.1.
Positive control:
n.a.
Observations and examinations performed and frequency:
CAGE SIDE OBSERVATIONS: Yes
- Time schedule: Throughout the study, all animals were checked early in each working day and again in the afternoon. At weekends and Public Holiday, a similar procedure was followed except that the final check was carried out at approximately mid-day. This allowed post mortem examinations to be carried out during the working period of that day

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: Once before commencement of treatment and at least once daily during treatment, each animal was observed, and any clinical signs recorded. Observations were performed at the same time interval each day; the interval was selected taking into consideration the presence of post-dose reactions.

BODY WEIGHT: Yes
- Time schedule for examinations: Each animal was weighed on the day of allocation to treatment group, on the day that treatment commenced, twice weekly thereafter and just prior to necropsy.

FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study):
- Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: Yes
- Time schedule: The weight of food consumed by each cage of rats was recorded at weekly intervals following allocation. The group mean daily intake per rat was calculated

HAEMATOLOGY: Yes
- Time schedule for collection of blood: End of week 2
- Anaesthetic used for blood collection: Yes (isofluorane)
- Animals fasted: Yes
- How many animals: all surviving male and female rats from each group
- Parameters checked in table No.1 in box “Any other information on materials & methods” were examined.

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: End of week 2
- Animals fasted: Yes
- How many animals: all surviving male and female rats from each group
- Parameters checked in table No.2 in box “Any other information on materials & methods” were examined.

OTHER: Organ weights: spleen, testes, thymus (where present)
Sacrifice and pathology:
GROSS PATHOLOGY: Yes, see table 3 in box "Any other information on materials & methods incl. tables"

HISTOPATHOLOGY: No
Statistics:
For continuous variables the significance of the differences amongst group means was assessed by Dunnett’s test or a modified t test, depending on the homogeneity of data.
Clinical signs:
effects observed, treatment-related
Description (incidence and severity):
Clinical signs in the surviving animals were limited to decreased activity and prone posture, observed on Day 6 in a single male animal of the high dose group (1000 mg/kg/day).
Lethargy, piloerection and prone posture were seen in one female of the same group on Day 11.
Mortality:
mortality observed, treatment-related
Description (incidence):
Two male animals of the high dose group, were found dead on Day 11 of the study. One animal was cannibalised by cage mates, while no signs were oberved during the in vivo phase in athe other animal. Post mortem macroscopic examination showed enlarged liver and distended urinary bladder in the latter animal. These changes were not considered to clearly establish the cause of death.
Body weight and weight changes:
effects observed, treatment-related
Description (incidence and severity):
Very slight but statistically significant reductions in body weight were seen in the high dose males on Days 8 and 11 (-6% and -7%, respectively). No significant differences were observed in the females.
Food consumption and compound intake (if feeding study):
effects observed, treatment-related
Description (incidence and severity):
Food consumption was slightly reduced in the high dose males on Days 8 and 15 (-14% and -22%), respectively when compared to controls. No significant differences in food consumption were observed in the females.
Food efficiency:
not examined
Water consumption and compound intake (if drinking water study):
not examined
Ophthalmological findings:
not examined
Haematological findings:
effects observed, non-treatment-related
Description (incidence and severity):
When compared to controls, erythrocytes were slightly higher in males dosed at 1000 mg/kg/day (9%). Due to the minimal severity and direction, this change was considered of no toxicological significance.
In addition, platelets were increased in males dosed at 100 and 300 mg/kg/day (40% and 48%, respectively) and decreased in one female receiving 300 mg/kg/day (95%). Due to the absence of dose-relation, those findings were considered incidental.
Coagulation: No changes were recorded.
Clinical biochemistry findings:
effects observed, non-treatment-related
Description (incidence and severity):
When compared to controls, glucose was increased in animals dosed at 1000 mg/kg/day (86% in males, 50% in females). The severity of this finding and the absence of other related changes were not considered to be suggestive of tissue/organ injury.
Other fluctuations were recorded, such as: decrease of urea in males dosed at 100 and 300 mg/kg/day (22% and 23%, respectively), increase of calcium in those receiving 300 and 1000 mg/kg/day (4%, both groups) and increase of alanine aminotransferase and aspartate aminotransferase in one male (300 mg/kg/day, 3.4 and 2.6-fold, respectively). Due to the absence of dose-relation and/or the minimal severity, these findings were considered of no toxicological relevance.
Urinalysis findings:
not examined
Behaviour (functional findings):
not examined
Immunological findings:
not examined
Organ weight findings including organ / body weight ratios:
no effects observed
Description (incidence and severity):
No changes of toxicological significance were seen in the weight of the organs.
Gross pathological findings:
effects observed, non-treatment-related
Description (incidence and severity):
Animals killed at termination did not show any relevant macroscopic changes.
The changes observed such as multiple red areas in the thymus are suggested to be incidental in untreated Sprague Dawley SD rats of the same age.
Neuropathological findings:
no effects observed
Histopathological findings: non-neoplastic:
not examined
Histopathological findings: neoplastic:
not examined
Other effects:
no effects observed
Details on results:
Two male animals dosed at 1000 mg/kg/day were found dead on Day 11 of the study. One of the 2 animals was cannibalised by cage mates. No signs were observed during the in vivo phase in the second one. Changes observed at post mortem observations were not considered to clearly establish the cause of death. Clinical signs (lethargy/decreased activity, prone posture and/or piloerection) were occasionally observed in individual animals dosed at 1000 mg/kg/day. Slight reductions in body weight, terminal body weight and food consumption were also observed during the in vivo phase in the males dosed at 1000 mg/kg/day. Increases in glucose and other minor fluctuations of clinical chemistry parameters were seen in animals dosed at 1000 mg/kg/day. No changes of toxicological significance were observed at post mortem observations (organ weights and macroscopic observations).

For summary of relevant finding see table 4 in box "Any other information on results incl. tables".
Key result
Dose descriptor:
NOEL
Effect level:
300 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
clinical signs
mortality
body weight and weight gain
food consumption and compound intake
Critical effects observed:
not specified

Table 4: Summary of relevant findings in the 2 -week study with CAPA 2043

Males Females
mg/kg bw/day 0 100 300 1000 0 100 300 1000
n 4 4 4 4 4 4 4 4
Mortality - - - 2 - - - -
Clinical signs - - - 1 animal: Cannibalized, Muzzle; Cannibalized, Ventrum
1 animal: Decreased activity, slight prone
- - - 1 animal: Lethargy, Piloerection, Moderate Prone
1 animal: Abrasion, Tail, Slight
Mean body weight (g) Dosing Day 1/4/8/11/15 185.98/209.28/236.00/254.10/259.63 186.43/208.70/237.60/261.50/266.95 187.00/210.00/240.80/264.35/268.73 143.03/186.08/201.90/221.03*D/235.70/241.75 150.88/159.83/172.88/185.18/180.38 147.20/157.65/170.65/184.28/181.10 147.48/156.48/168.73/180.73/179.98 146.33/154.13/165.33/175.75/173.88
Mean food consumption (g) Dosing Day 6/8/15 19.46/20.09/17.63 19.25/20.66/18.44 19.33/20.87/18.99 19.25/17.29/13.78 15.81/13.72/12.48 14.32/13.79/12.33 14.26/14.13/12.43 13.81/13.11/11.84
Haematology
Erythrocytes (x10^6/µL) 6.890 6.738 6.913 7.515*D 7.103 6.878 7.270 7.203
Platelets (x10^3/µL) 777.7 1089.0* 1151.3*D 940.0 1114.3 1124.5 783.0 1109.3
Clinical Chemistry
Glucose (mg/dL) 60.23 49.55 63.83 111.90+D 62.90 65.50 87.78 94.23*D
Ca (mMmol/L) 2.535 2.598 2.638+D 2.635*D 2.578 2.565 2.553 2.650
Urea (mg/dL) 33.63 26.28+D 25.83+D 30.30 36.60 36.38 38.43 32.15
Alanine aminotransferase (U/L) 41.18 43.73 66.58 64.15 33.58 30.60 34.20 33.70
Aspartate aminotransferase (U/L) 91.83 84.80 120.85 73.35 84.08 67.48 69.38 67.90
Gross pathological findings
1 animal: Thymus red - left lobe 1 animal: liver: single, pale 1 animal: Thymus multiple red - right lobe 1. animal (unscheduled death; cannibalised animal): Thymus: Multiple, Dark red
2. animal /unscheduled death): Epididymis small, liver enlarged, prostate gland small, seminal vesicles small, urinary bladder distended)
1 animal (final sacrifice): Thymus multiple red - right lobe
- 1 animal: Uterus distended, clear, fluid - -

* Dunnett LSD Test Significant at the 0.05 level

+D Dunnett LSD Test Significant at the 0.01 level

Conclusions:
In conclusion, the test item CAPA 2043, when orally administered at dose levels of 100, 300 and 1000 mg/kg/day, resulted to be satisfactorily tolerated by Sprague Dawley rats at the low and intermediate dose levels (100 and 300 mg/kg/day), while the highest dose level of 1000 mg/kg/day induced some toxicity (mortality or occasional clinical signs, slight reductions in body weight and food consumption). Therefore, it can be concluded that 300 mg/kg/day was the NOEL (No Observed Effect Level) in this study. The highest dose of 1000 mg/kg/day was not considered to be safe in a subsequent toxicity study of longer duration in Sprague Dawley rats.
Executive summary:

In a 2-week repeated dose toxicity study, the test item CAPA 2043 (2-oxepanone, polymer with 1,4-butanediol) in corn oil was administered to Sprague-Dawley rats (4/sex/dose) by gavage at dose levels of 0, 100, 300 and 1000 mg/kg bw/day for once a day, 7 days a week for 2 weeks.

Two male animals dosed at 1000 mg/kg/day were found dead on Day 11 of the study. One of the 2 animals was cannibalised by cage mates. No signs were observed during thein vivophase in the second one. Changes observed at post mortem observations were not considered to clearly establish the cause of death. Clinical signs (lethargy/decreased activity, prone posture and/or piloerection) were occasionally observed in individual animals dosed at 1000 mg/kg/day. Slight reductions in body weight, terminal body weight and food consumption were also observed during thein vivophase in the males dosed at 1000 mg/kg/day. Increases in glucose and other minor fluctuations of clinical chemistry parameters were seen in animals dosed at 1000 mg/kg/day. No changes of toxicological significance were observed atpost mortemobservations (organ

weights and macroscopic observations). In conclusion, the test item CAPA 2043, when orally administered at dose levels of 100, 300 and 1000 mg/kg/day, resulted to be satisfactorily tolerated by Sprague Dawley rats at the low and intermediate dose levels (100 and 300 mg/kg/day), while the highest dose level of 1000 mg/kg/day induced some toxicity (mortality or occasional clinical signs, slight reductions in body weight and food consumption). Therefore, it can be concluded that 300 mg/kg/day was the NOEL (No Observed Effect Level) in this study. The highest dose of 1000 mg/kg/day was not considered to be safe in a subsequent toxicity study of longer duration in Sprague Dawley rats.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2017
Report Date:
2017

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 414 (Prenatal Developmental Toxicity Study)
Version / remarks:
22 January 2001
Deviations:
no
GLP compliance:
yes
Limit test:
no

Test material

Reference
Name:
Unnamed
Type:
Constituent
Type:
Constituent
Details on test material:
Trade name: Capa 2043
Chemical name: 2-oxepanone, polymer with 1,4-butanediol
CAS number: 31831-53-5
EC number: 608-670-1
Batch number: WAC 000392
Carbon content (TOC): 61%
Appearnce: Clear colourless liquid
Vapour pressure : <1 Pa at 25oC
Density 20oC: 1.05 g/cm3
Stability: stable at normal conditions
Storage conditions: Sealed containers, dark
Specific details on test material used for the study:
Identity: CAPA 2043
Label name: CAPA(TM) 2043
Chemical name 2-oxepanone, polymer with 1,4-butanediol
Batch no. WAC000478
Purity: 100 % (UVCB)
CAS number 31831-53-5
EC number 608-670-1
Expiry date 10 May 2018
Storage conditions Cool, dry conditions in well-scaled receptacles
Appearance Clear, colourless liquid

Test animals

Species:
rat
Strain:
Sprague-Dawley
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Envigo RMS srl, San Pietro al Natisone (UD), Italy
- Age at arrival: at least 11 weeks old
- Weight at arrival: 202-219 g for females and 306-344 g for males
- Housing: Before and after the pairing period, the animals were housed no more than 5 of one sex to a cage in polysulfone solid bottomed cages. Nesting material was provided inside suitable bedding bags and changed at least 3 times a week. During the pairing period, the rats were housed on the basis of 1 male to 1 female in clear polysulfone cages with a stainless steel mesh lid and floor. Each cage tray held absorbent material which was inspected and changed daily.
- Diet: commercially available laboratory rodent diet (4 RF 21, Mucedola S.r.l., Via G. Galilei, 4, 20019 Settimo Milanese (MI), Italy); ad libitum
- Water: drinking water; ad libitum
- Acclimation period: 19 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 °C ± 2 °C
- Humidity (%): 55% ± 15%
- Air changes (per hr): 15-20
- Photoperiod (hrs dark / hrs light): 12 hrs dark/12 hrs light

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
corn oil
Details on exposure:
The test item was administered orally by gavage at a dose volume of 4mL/kg bw. Control animals received the vehicle alone at the same dose volume. The dose was administered to each animal on the basis of the most recently recorded body weight and the volume administered was recorded for each animal.

PREPARATION OF DOSING SOLUTIONS: The required amount of CAPA 2043 was suspended in the vehicle. The formulation was prepared daily (concentrations of 50, 100 and 200 mg/mL). Concentrations were calculated and expressed in terms of test item as supplied.

VEHICLE: corn oil
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
Prior to commencement of treatment, analysis was performed to confirm that the proposed formulation procedure was acceptable. Results of the analyses were within the limits of acceptance. Stability at 3 and 8 days at room temperature was verified in the range from 10 to 120 mg/mL in RTC Study No. A2242. In addition, the stability was verified in this study at 200 mg/mL and found to be 28 hours and 8 days at room temperature. Samples of the formulations prepared at Week 1 and Week 3 of the study were also analysed to check the concentration and homogeneity. Results of the analyses were within the limits of acceptance. Chemical analysis was carried out by the laboratory, according to a validated method (RTC Study no. A2242 with validation in the range from 10 to 120 mg/mL and the present RTC Study no. X0460 with validation up to 200 mg/mL.
Details on mating procedure:
- Impregnation procedure: cohoused
- If cohoused: Females were paired 1 to 1 in the home cage of the male and left overnight.
- M/F ratio per cage: 1:1
- Length of cohabitation: overnight; vaginal smears were taken daily in the morning from the day after pairing until a positive identification of mating was made
- Proof of pregnancy: sperm in vaginal smear referred to as day 0 of pregnancy
- Any other deviations from standard protocol: no
Duration of treatment / exposure:
Gestation Day 6-19
Frequency of treatment:
Once daily (GD 6-19)
Duration of test:
The animals were euthanised on Gestation Day 20.
Doses / concentrationsopen allclose all
Dose / conc.:
0 mg/kg bw/day
Remarks:
Vehicle (corn oil) control
Dose / conc.:
200 mg/kg bw/day
Remarks:
Low dose group
Dose / conc.:
400 mg/kg bw/day
Remarks:
Mid dose group
Dose / conc.:
800 mg/kg bw/day
Remarks:
High dose group
No. of animals per sex per dose:
Each group comprised 24 mated female rats.
Control animals:
yes, concurrent vehicle
Details on study design:
- Dose selection rationale: Dose levels were selected in consultation with the Sponsor based on information from a preliminary non GLP compliant study (RTC Study No. E0064, see IUCLID section 7.5.1, Oberto G. (2017)). This oral gavage preliminary study was carried out at dose levels of 100, 300 and 1000 mg/kg bw/day. In this study, the test item CAPA 2043 was satisfactorily tolerated by Sprague Dawley rats at the low and intermediate dose levels (100 and 300 mg/kg bw/day), while the highest dose level of 1000 mg/kg bw/day induced some toxicity (mortality or occasional clinical signs, slight reductions in body weight and food consumption). A NOEL of 300 mg/kg bw/day was determined in this preliminary study.
As the highest dose in the preliminary study (1000 mg/kg bw/day) is not considered to be safe in a subsequent toxicity of longer duration in Sprague Dawley rats, a lower dose was chosen which is higher than the NOEL of 300 mg/kg bw/day. Thus, a high dose of 800 mg/kg bw/day was used, with the aim to induce some developmental and/or maternal toxicity (clinical signs or a decrease in body weight) but not death or severe suffering, as required in the OECD guideline 414.
Furthermore, according to OECD guideline 414, at least one intermediate dose level should produce minimal observable toxic effects. The lowest dose level should not produce any evidence of either maternal or developmental toxicity. A descending sequence of dose levels should be selected with a view to demonstrating any dosage-related response and no-observed-adverse-effect level (NOAEL) or doses near the limit of detection that would allow the determination of a benchmark dose. It is stated that two to four-fold intervals are frequently optimal for setting the descending dose levels. Since a NOAEL of 300 mg/kg bw/day was determined in the 2-week preliminary study, a mid-dose of 400 mg/kg bw was chosen for the prenatal developmental study and a low dose of 200 mg/kg bw/day was selected based on the recommendations of the guideline.
- Rationale for animal assignment: On the day of allocation (Day 0 post coitum) all females were weighed and allocated to the groups by computerised stratified randomisation to give approximately equal initial group mean body weights. Each female was identified within the study by ear notch and housed no more than 4 to a cage. The cages were identified by a label recording the study number, animal numbers and details of treatment. The arrangement of cages in batteries was such that cages from each treatment group were evenly distributed across the battery to minimize possible environmental effects.
The group identification and animal numbers assigned to the treatment are summarized below:

Group 1 (0 mg/kg bw/day): Females numbers (odd only): 1-47
Group 2 (200 mg/kg bw/day): Females numbers (odd only): 49-95
Group 3 (400 mg/kg bw/day): Females numbers (odd only): 97-143
Group 4 (800 mg/kg bw/day): Females numbers (odd only): 145-191

The rat numbers listed above formed the last digits of a computer generated 8 figure animal number (the remaining digits of the animal number were different for each concurrent study and served to ensure unique animal numbering for any study employing computerised data collection). The computerised system used in this study was the Xybion Path/Tox System, Version 4.2.2.

Examinations

Maternal examinations:
CAGE SIDE OBSERVATIONS: Yes
- Time schedule: Throughout the study, all animals were checked early in each working day and again in the afternoon. At weekends and Public Holidays, a similar procedure was followed except that the final check was carried out at approximately mid-day.

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: clinical signs were recorded for individual animals. Each animal was observed twice daily during treatment before dosing and 30 minutes -1 hour after dosing and any clinical signs recorded starting from allocation until sacrifice.

BODY WEIGHT: Yes
- Time schedule for examinations: Days 0, 3, 6, 9, 12, 15, 18 and 20 post coitum

FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study): Yes
- Time schedule for examinations: Days 0, 3, 6, 9, 12, 15, 18 and 20 post coitum
- Cages with at least one not pregnant female were excluded from group mean calculation
- Compound intake calculated as time-weighted averages from the consumption and body weight gain data: No (no feeding study)

WATER CONSUMPTION AND COMPOUND INTAKE (if drinking water study): No

POST-MORTEM EXAMINATIONS: Yes
- Sacrifice on Gestation day 20
- Organs examined: Ovaries and uteri
Ovaries and uterine content:
The ovaries and uterine content was examined after termination: Yes
Examinations included:
- Gravid uterus weight: Yes
- Number of corpora lutea: Yes
- Number of implantations: Yes
- Number of early resorptions: Yes
- Number of late resorptions: Yes
- Other: number, sex and weight of all live foetuses; number and sex of dead foetuses (foetuses at term without spontaneous movements and breathing); number of intra-uterine deaths; gross evaluation of placentae
Fetal examinations:
- External examinations: Yes: all per litter
- Soft tissue examinations: Yes: approx. half per litter
- Skeletal examinations: Yes: approx. half per litter
- Head examinations: Yes: no data

Skeletal and fixed-visceral examinations were performed in all groups. Structural deviation was classified as follows:
- Malformations: Major abnormalities that are rare and/or affect survival or health.
- Anomalies: Minor abnormalities that are detected relatively frequently.
- Variants: A change that occurs within the normal population under investigation and is unlikely to adversely affect survival or health. This might include a delay in growth or morphogenesis that would have otherwise followed a normal pattern of development.
Statistics:
For continuous variables the significance of the differences amongst group means was assessed by Dunnett’s test or a modified t-test, depending on the homogeneity of data. Statistical analysis of non-continuous variables was carried out by means of the Kruskal-Wallis test and intergroup differences between the control and treated groups assessed by a non-parametric version of Williams test.
Indices:
Pre-implantation loss was calculated as a percentage from the formula: Pre-implantation Loss (%) = [no. corpora lutea − no. implantations / no. corpora lutea] ×100

Post-implantation loss was calculated as a percentage from the formula: Post implantation Loss (%) = [no. implantations − no. live foetuses / no. implantations] ×100

Total implantation loss was calculated as a percentage from the formula: Total implantation Loss (%) = [no. corpora lutea − no. live foetuses / no. corpora lutea] ×100

Sex ratios of the foetuses were calculated as the percentage of males per litter. All derived values (e.g., means, percentages, ratios) were first calculated within the litter and the group values derived as a mean of individual litter values. Foetal structural deviations were expressed as the percentage of affected foetuses relative to all foetuses examined per group, as well as in terms of the mean litter percentage of affected litters.
Historical control data:
No data

Results and discussion

Results: maternal animals

General toxicity (maternal animals)

Clinical signs:
effects observed, non-treatment-related
Description (incidence and severity):
Signs of reaction to treatment were limited to salivation observed during the dosing period in 11 out of 24 females of the high-dose group starting on GD8 post coitum and in 1 out of 24 females of the low dose group on GD 13. Hairloss of the lower forelimb was also detected in one female of the high-dose group at the end of treatment. These signs were not to be considered of toxicological significance.
Dermal irritation (if dermal study):
not examined
Mortality:
no mortality observed
Description (incidence):
No animals died during the study.
Body weight and weight changes:
effects observed, non-treatment-related
Description (incidence and severity):
No differences in body weight were noted between the control and treated groups. A significant decrease in body weight gain of approximately 11% was noted on GD20 in females of the low-dose group when compared to controls. This change was considered to be without toxicological relevance in the absence of findings in other dose groups.
Food consumption and compound intake (if feeding study):
no effects observed
Description (incidence and severity):
No changes were detected in food consumption between treated and control females.
Food efficiency:
not examined
Water consumption and compound intake (if drinking water study):
not examined
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:
no effects observed
Description (incidence and severity):
No significant differences in terminal body weight, gravid uterus weight and absolute weight gain were observed in treated groups compared to the control group.
Gross pathological findings:
effects observed, non-treatment-related
Description (incidence and severity):
No treatment-related changes were noted. The sporadic changes, such as marked hairloss of the forelimbs in one high-dose female or thickened mucosa, white firm contents of the urinary bladder and distended right ureter in one low-dose female, were not attributable to treatment.
Neuropathological findings:
not examined
Histopathological findings: non-neoplastic:
not examined
Histopathological findings: neoplastic:
not examined
Details on results:
Effects were observed in clinical signs (hairloss, salivation), as well as in body weight, food consumption, gravid uterus weight and macroscopic observation of treated females when compared to controls. These effects were not considered to be treatment-related.

Maternal developmental toxicity

Number of abortions:
no effects observed
Description (incidence and severity):
No abortions were reported.
Pre- and post-implantation loss:
effects observed, non-treatment-related
Description (incidence and severity):
In the control group one female showed unilateral implantation with total resorption. Unilateral implantation was also detected in one female of the low dose group.
Total litter losses by resorption:
no effects observed
Description (incidence and severity):
No effects on total litter losses by resorption were reported.
Early or late resorptions:
effects observed, non-treatment-related
Description (incidence and severity):
In the control group one female showed unilateral implantation with total resorption.
Dead fetuses:
no effects observed
Description (incidence and severity):
No dead fetuses were observed.
Changes in pregnancy duration:
not examined
Changes in number of pregnant:
effects observed, non-treatment-related
Description (incidence and severity):
A total of 10 females were found not pregnant at necropsy: one in the control group, four in the low-dose group, two in the mid-dose group and three in the high-dose group.
Other effects:
not examined
Details on maternal toxic effects:
In the control group one female showed unilateral implantation with total resorption. Unilateral implantation was also detected in one female of the low dose group. A total of 10 females were found not pregnant at necropsy: one in the control group, four in the low-dose group, two in the mid-dose group and three in the high-dose group. These effects were considered to be non-treatment related, since no dose-response relationship was observed.

Effect levels (maternal animals)

Key result
Dose descriptor:
NOAEL
Effect level:
800 mg/kg bw/day
Based on:
test mat.
Basis for effect level:
other: no adverse effects observed

Maternal abnormalities

Key result
Abnormalities:
no effects observed

Results (fetuses)

Fetal body weight changes:
no effects observed
Description (incidence and severity):
Mean foetal weight was not affected by treatment.
Reduction in number of live offspring:
no effects observed
Description (incidence and severity):
A reduction in the number of live offspring was not reported.
Changes in sex ratio:
no effects observed
Description (incidence and severity):
Sex ratio was not affected by treatment.
Changes in litter size and weights:
no effects observed
Description (incidence and severity):
Litter data were not affected by treatment.
Changes in postnatal survival:
not examined
External malformations:
effects observed, non-treatment-related
Description (incidence and severity):
A total of 2 small foetuses (< 2.7 g) were detected, both in the mid-dose group. No abnormalities were detected in the control, low- or high-dose groups.
Skeletal malformations:
no effects observed
Description (incidence and severity):
No changes were noted at the skeletal examination of the foetuses which were considered to be treatment-related.
Visceral malformations:
effects observed, non-treatment-related
Description (incidence and severity):
No changes were seen at the visceral examination of the foetuses which were considered to be treatment-related. Extreme pelvic dilatation and/or extremely enlarged ureter (malformations) were seen in a single foetus each in the low- and the mid-dose groups. These isolated cases were considered without toxicological relevance. Anomalies were seen in treated as well as control groups, with similar (testes displaced) or higher (generalised oedema, ureter moderately enlarged) incidence in the control group and, therefore, considered toxicologically unremarkable.
Other effects:
no effects observed
Details on embryotoxic / teratogenic effects:
No changes were noted at the visceral foetuses examination and in relation to external malformations which were considered treatment-related.
There was no evidence of teratogenicity or developmental toxicity in this study.

Effect levels (fetuses)

Key result
Dose descriptor:
NOAEL
Effect level:
800 mg/kg bw/day
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: no adverse effects observed

Fetal abnormalities

Key result
Abnormalities:
no effects observed

Overall developmental toxicity

Key result
Developmental effects observed:
no

Any other information on results incl. tables

Table 1: Summary of relevant findings in the prenatal developmental toxicity study with CAPA 2043

Dose level (mg/kg bw/day) 0 200 400 800
Mated (#) 24 24 24 24
Deaths (#) - - - -
Not pregnant (#) 1 4 2 3
Dams with abortions (#) - - - -
Dams with early deliveries (#) - - - -
Dams with stillbirths (#) - - - -
Dans wutg dead foetuses (#) - - - -
Unilateral implantation and total resorption 1 0 0 0
Unilateral implantation 0 1 0 0
Dams with live fetuses on GD20 (#) 22 20 22 21
Clinical signs of females - Salivation (1/24 animals/4.2% of animals with observation during interval) - Hairloss (1/24 animals/4.2% of animals with observation during interval)
Salivation (11/24 animals/45.8% of animals with observations during interval)
Mean body weight (g) GD0 232.71 231.74 228.83 231.76
GD6 256.02 256.69 254.23 256.15
GD9 265.36 265.09 263.15 265.81
GD18 342.80 341.83 343.49 336.61
GD20 375.06 370.55 375.04 367.21
Mean body weight gain (g/day) GD6 3.422 3.652 3.367 3.269
GD9 3.115 2.832 2.974 3.222
GD20 16.132 14.363* 15.778 15.299
Mean body weight gain (g) GD6-9 9.34 8.40 8.92 9.66
GD6-20 119.04 113.86 120.81 111.06
GD0-20 142.35 138.81 146.21 135.45
Mean gravid uterus weight (g) 75.33 71.12 74.24 71.54
Mean absolute weight gain (g) 59.62 65.90 68.89 60.55
Corpora lutea (#) 13.14 12.45 12.77 12.76
Implantations (#) 13.00 12.35 12.45 12.62
Pre-implantation loss (%) 1.02 0.67 2.66 1.07
Post-implantation loss (%) 3.54 1.60 0.86 4.64
Total implantation loss (%) 4.51 2.27 3.52 5.68
Mean viable foetuses (#) 12.55 12.15 12.32 12.05
Viable foetuses (%) 100 100 100 100
Mean foetal weight (g) (sexes combined) 3.91 3.86 3.92 3.94
Mean foetal weight (g) by sex not reported
Small foetuses (#) - - 2 -
Mean litter weight (g) 48.92 46.86 48.17 47.24
Early uterine deaths (%) 0.45 0.20 0.14 0.57
Late uterine deaths (%) 0.00 0.00 0.00 0.00
Males (%) 51.37 50.19 47.73 46.41
External abnormalities in foetuses (% of foetuses) - - 0.74 -
External abnormalities in foetuses (# of foetuses) - - 2 -

*significantly different to controls (p<0.05)

Applicant's summary and conclusion

Conclusions:
In this study, maternal and developmental NOAELs of 800 mg/kg bw/day can be considered.
Executive summary:

In a developmental toxicity study conducted according to OECD guideline 414, CAPA 2043 (2-oxepanone, polymer with 1,4-butanediol diluted in corn oil) was administered to 24 pregnant female Sprague-Dawley rats/dose by gavage at dose levels of 0, 200, 400 or 800 mg/kg bw/day from days 6 through 19 of gestation. Rats were terminated on gestation day 20 and the uterine contents investigated. Foetuses were assessed for external, visceral and skeletal abnormalities.

No maternal deaths occurred during the study. Clinical signs of treatment were limited to post-dosing salivation in a number of maternal rats at the highest dose level, which is not considered to be of toxicological significance. A reduction of the body weight in the low-dose group is considered to be not treatment-related, since no dose-response relationship was observed. Food consumption was unaffected by treatment. Litter parameters were unaffected by treatment. Litter size and foetal weight were comparable in all groups. There were no changes in the incidence or nature of foetal external, visceral or skeletal findings indicative of an effect of treatment. In the absence of any toxicologically significant maternal effects and in the absence of any developmental toxicity, maternal and developmental NOAELs of 800 mg/kg bw/day can be determined for this study.

The developmental toxicity study in the rat is classified acceptable and satisfies the guideline requirement for a developmental toxicity study (OECD 414) in rats.