N-(2-hydroxypropyl)oleamide

Developmental toxicity / teratogenicity

Administrative data

Endpoint:

developmental toxicity

Type of information:

experimental study

Adequacy of study:

key study

Study period:

November 2016 - June 2017

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2018

Report date:

2018

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 414 (Prenatal Developmental Toxicity Study)

Deviations:

no

Remarks:

no major deviations

GLP compliance:

yes (incl. QA statement)

Remarks:

23 march 2017

Limit test:

no

Test material

Test material information

Constituent 1

Reference substance name:

N-(2-hydroxypropyl)oleamide

EC Number:

203-828-2

EC Name:

N-(2-hydroxypropyl)oleamide

Cas Number:

111-05-7

Molecular formula:

C21H41NO2

IUPAC Name:

N-(2-hydroxypropyl)oleamide

Test material form:

solid

Remarks:

beige waxy solid

Details on test material:

LOT/BATCH: 160602013073 w/o solvent

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: 160602013073 w/o Solvent
- Expiration date of the lot/batch: 2018/05/30
- Purity test date: 2016/06/22

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material:
Immediately upon receipt, the test item was registered, then stored at room temperature in accordance with the Sponsor's instructions. The complete description of the chemical and physical properties of the test item including stability is the responsibility of the Sponsor.
- Solubility and stability of the test substance in the solvent/vehicle:
the test item formulations at 20 mg/mL to 200 mg/mL in its vehicle are stable for 30 days at room temperature

TREATMENT OF TEST MATERIAL PRIOR TO TESTING
- Final preparation of a solid:
The test item was warmed in a water bath at approximately 50°C. An appropriate amount of test item was weighed. Corn oil previously warmed in water bath at approximately 50°C was added up to obtain the appropriate amount. The formulation was mixed using magnetic stirrer in the water bath at approximately 50°C for 10 minutes. Then the formulation was mixed using magnetic stirrer at room temperature for at least 30 minutes
- Form of administration: On the day of treatment, the formulation and vehicle were warmed using a water bath at approximately 50°C under magnetic stirring for at least 30 minutes, then they were kept under magnetically stirring at ambient temperature for at least 30 minutes. The vehicle was warmed at 40°C on 16 Dec 2016. N-(2-hydroxypropyl) Oleamide was administered as a suspension in its vehicle. Control animals were given the vehicle under the same conditions as animals dosed with N-(2-hydroxypropyl) Oleamide. The test item formulations were prepared and kept at room temperature and used within one week after preparation.

Test animals

Species:

rat

Strain:

Sprague-Dawley

Remarks:

SPF (Specific Pathogen Free) Sprague-Dawley - Crl: OFA (SD)

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River Laboratories France, Domaine des Oncins, 69210 L'ARBRESLE Cedex, France
- Age at study initiation: 10 to 11 weeks at the beginning of the treatment period
- Weight at study initiation: Between 224.4 and 326.7g on the day of randomisation (D5pc) The weight variation of animals used was minimal (+/- 20% of the mean weight). About 10% more animals were ordered to allow selection of animals according to the criterion of body weight and they were used as spare animals in case of unforeseen events happen
- Fasting period before study:
- Housing: Animals were housed individually in cages of standard dimensions with sawdust bedding.
- Diet (e.g. ad libitum): RM3 (E)-SQC SDS/DIETEX feed (quality controlled/radiation sterilised) was available ad libitum except during the fasting experimental period. The certificate of analysis concerning this feed product is included in the report. The criteria for acceptable levels of contaminants in the feed supply were within the limits of the analytical specifications established by the diet manufacturer.
- Water (e.g. ad libitum): Drinking water was available ad libitum in polycarbonate bottles with a stainless steel nipple. A specimen of water is obtained approximately every 6 months and sent to Laboratoire de la Touraine - ZA n°1 du Papillon - Rue de l0Aviation - 37210 Parçay-Meslay - France, for analysis. The certificates of analysis are included in the report. The criteria for acceptable levels of contaminants in the water supplied were within the limits of the analytical specifications.
- Acclimation period:Animals arrived at CERB on Day 1 of pregnancy (D1pc). Animals were supplied in several batches for logistical reasons. Each animal had five days in the laboratory animal house where the experiment took place before beginning of dosing. Only animals without any visible sign of illness were used for the study.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): The animals were placed in an air-conditioned (20-24°C) animal house
- Humidity (%): relative humidity between 45% and 65% (except during the cleaning slot). Between 28 Nov at 07.35 p.m. up to 29 Nov at 11.10 a.m., an abnormal decrease of hygrometry was noted in animal room.
- Air changes (per hr): non-recycled filtered air was changed approximately 10 times per hour
- Photoperiod (hrs dark / hrs light): The artificial day/night cycle involved 12 hours light and 12 hours darkness with light on at 7.30 a.m

IN-LIFE DATES: From: 25 November 2016 To: 22 December 2016

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

corn oil

Details on exposure:

DIET PREPARATION no information available

VEHICLE
Corn oil will be used (Reference C8267)
- Lot/batch no. (if required): MKBS6944V and MKBW9504V, Expiry dates: 08 Oct 2020 and 06 Oct 2021 respectively
- Amount of vehicle (if gavage): 5 mL/kg body weight.

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

Concentrations of test item in formulations were checked during the first and last week of the study.
Each concentration level and the vehicle were checked.

1 mL samples of each test item dosing formulation were taken in duplicate by top, middle and bottom sampling. Similar samples from the vehicle were taken, from the middle of the formulation only. Only the middle samples were assayed. Samples were collected in glass ontainers and stored at room temperature.
Labels on the containers were marked in waterproof ink with Testing Facility, Study Number, Name of the test item and Concentrations, Sampling Date, Sampling Time and Storage conditions. The sample labels also indicated whether the samples were taken from the top, middle (for vehicle) or bottom of the formulation container.
The identity and concentration of the test item in the samples and the absence of the test item in the control formulation was determined by liquid chromatography with UV-detection within the validated stability period available at this moment i.e. one week after sampling Acceptance criteria of the formulation analysis were fixed usually +/- 15% to the nominal concentration.

The analytical method is validated within the range 85-115% of the theoretical concentration for formulations between 19.765 mg/mL and 198.120 mg/mL, with a precision better than 10%. The samples in corn oil must be analysed within the stability period (i.e. 30 days at room temperature) and must be within the range 85-115% to meet the acceptance criteria.

Formulation analysis was performed on one formulation prepared during the first and the last week of the study. The concentrations tested were 20 mg/mL, 60 mg/mL and 200 mg/mL. The concentrations found were within the range of acceptance (15% of the intended concentration). The absence of test
item was also confirmed in the vehicle samples.
Therefore, these data confirmed that the formulations were properly prepared.

Details on mating procedure:

- Impregnation procedure:
The breeding establishment will be responsible for mating. Females will be mated at the beginning of the morning. They will be inspected for the presence of a vaginal plug.

- Proof of pregnancy: vaginal plug referred to as day 0 of pregnancy (D0pc)

Duration of treatment / exposure:

The test item administered in pregnant rats from Day 6 to Day 19 of gestation

Frequency of treatment:

N-(2-hydroxypropyl) Oleamide or its vehicle will be administered once a day at approximately the same time (a maximum range of 4 hours between the start and the end of the daily treatment) at each chosen dose level

Duration of test:

21 days (Days 0-20 post coitum)

Doses / concentrations

Doses / concentrations 1

Dose / conc.:

100 mg/kg bw/day

Doses / concentrations 2

Dose / conc.:

300 mg/kg bw/day

Doses / concentrations 3

Dose / conc.:

1 000 mg/kg bw/day

No. of animals per sex per dose:

20 mated females/dose

Control animals:

yes, concurrent vehicle

Details on study design:

- Dose selection rationale: Proposed doses are selected in agreement with the Sponsor. The choice is based on previous studies (COMBINED REPEATED DOSE TOXICITY STUDY WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST BY ORAL ROUTE (GAVAGE) IN RATS - OECD 422 - Study No. 39644 RSR). Moreover, the highest dose should reveal signs of toxicity and the lowest dose should represent a no-observed-adverse effects level

Examinations

Maternal examinations:

CAGE SIDE OBSERVATIONS: No data

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: Animals were observed in the home cage before the first dosing and at least once a day during the study except on D7 for 3/20 female of group 2. The time of observation during the treatment period was at 60 min post dose (+/- 30 min). Females showing signs of abortion or of premature delivery during the study, the day on which such findings seen were noted.

BODY WEIGHT: Yes
- Time schedule for examinations: Animals were weighed on the following days:
• on D1 and D5 (day of randomisation)
• daily during treatment (from D6 to D19)
• on D20, day of necropsy, not fasted and not exsanguinated

FOOD CONSUMPTION : Yes
- Food consumption was measured and presented daily from D6 to D19

WATER CONSUMPTION : No data

POST-MORTEM EXAMINATIONS: Yes
- Sacrifice on gestation day # 20: On day 20 of pregnancy, all surviving animals were killed by subtotal exsanguination following isoflurane inhalation. Females showing signs of abortion or of premature delivery during the study were killed on the day of such findings
- Organs examined: Animals showing signs of abortion or of premature delivery were necropsied as quickly as possible and specimens as required by the study plan obtained whenever practically possible. The main organs cited below were examined macroscopically. All animals were subjected to gross necropsy and their organs (brain, liver, stomach, small intestine (duodenum, jejunum, ileum), large intestine (caecum, colon, rectum), kidneys, spleen, adrenals, lungs, heart) were examined macroscopically. Uterus and ovaries from each female were macroscopically observed and fixed in an appropriate fixative. Gravid uteri were weighed before extraction of foetuses

OTHER:
All organs showing macroscopic signs of pathology and corresponding organs of control groups for comparison were fixed in an appropriate fixative. Remaining tissues were destroyed 6 months after issue of the draft release.

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
- Number and uterine location of foetuses (live and dead): Yes

Fetal examinations:

- External examinations: Yes: [all per litter]
- Soft tissue examinations: No data
- Skeletal examinations: Yes: [performed in the first instance only on control and high dose groups]
- Number of live or dead foetuses: Yes
- Individual foetal body weight (live and dead): Yes : [all per litter]
- Caudo-cranial measurement of live and dead foetuses: Yes : [all per litter]
- Gross evaluation and weight of placenta of all foetuses: Yes : [all per litter]
- Sexing of all foetuses: Yes : [all per litter]

Statistics:

See "Any other information on materials and methods incl. tables"
The validated computerised system used in this phase was the Xybion Path/Tox System, Version 4.2.2.

Indices:

Pre-implantation loss and post-implantation loss were calculated according to the following formula:
Pre-implantation loss (%):
((Number of corpora lutea - number of implantations) / Number of corpora lutea) x 100
Post-implantation loss (%):
((Number of implantations - number of live foetuses) / Number of implantations) x 100

Foetal or litter incidence was calculated according to the following formula:
Foetal or litter incidence (%):
(Total number of foetus or litter with a particular finding / Total number of foetus per group) x 100

Historical control data:

No data available

Results and discussion

Results: maternal animals

General toxicity (maternal animals)

Clinical signs:

no effects observed

Description (incidence and severity):

There was an increased salivation in some females treated with test item at 300 mg/kg (in 1/20) on D13 and D14 and at 1000 mg/kg from D12 up to the end of the study (between 1 to 3/20). There was also chromodacryorrhoea in 1 or 2/20 different animals following the day of observation in females treated with test item at 300 mg/kg or at 1000 mg/kg. These signs were of low incidence and were not attributed to a toxicological effect of the test item.
There was aggressiveness on D5 in 1/20 female treated with test item at 100 mg/kg. This sign was only observed on the first day of treatment and was not attributed to the test item.

Mortality:

no mortality observed

Description (incidence):

No mortality and no relevant clinical signs or signs of reaction to treatment were noted in treated females.

Body weight and weight changes:

no effects observed

Description (incidence and severity):

There was no change in body weight gain.

Food efficiency:

no effects observed

Description (incidence and severity):

There was a statistically significant isolated lower food consumption in females treated with test item at 100 mg/kg on D10 (-13%) and D18 ( -20%) or in females treated with test item at 1000 mg/kg on D6, D11 or D18 (between -14 and -18%). This lower food consumption is transient and attributable to
the high variability in the control group.
From D15pc, food consumption was lower for Female No. 1602737 treated with test item at 100 mg/kg.

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):

no effects observed

Immunological findings:

not examined

Organ weight findings including organ / body weight ratios:

no effects observed

Description (incidence and severity):

There was no change in uterus weight.

Gross pathological findings:

no effects observed

Description (incidence and severity):

There were no macroscopic findings in mated females killed on D20pc.

At the necropsy, there was a dark liquid in the vulva and vagina, a black abnormal content in stomach and enlarged spleen in Female No. 1602755 treated with test item at 300 mg/kg.
In Female No. 1602730 treated with test item at 1000 mg/kg, there was dark liquid in the vagina, transparent and abnormal area in stomach, adhesion between lungs and thoracic cavity, adhesion between lungs, heart and diaphragm, cloudy liquid in the thoracic cavity and heart with firm area and granular aspect

Neuropathological findings:

not examined

Histopathological findings: non-neoplastic:

not examined

Histopathological findings: neoplastic:

no effects observed

Other effects:

no effects observed

Maternal developmental toxicity

Number of abortions:

effects observed, non-treatment-related

Description (incidence and severity):

Signs of abortion (blood near genital orifice and in cage) were seen in two females, on D19 in one female treated with the intermediate dose (No. 1602755) and on D16 in one female treated with the highest dose (No. 1602730)

Pre- and post-implantation loss:

effects observed, non-treatment-related

Description (incidence and severity):

There was a higher percentage of pre and post implantation loss for Female No. 1602737 treated with test item at 100 mg/kg and Female No. 1602766 treated with test item at 300 mg/kg. For these females, there was no foetus, only resorptions were observed

Total litter losses by resorption:

no effects observed

Early or late resorptions:

effects observed, non-treatment-related

Description (incidence and severity):

In Female No. 1602755, there is 16 implantation sites, with 1 early resorption.
In Female No. 1602730, there is 17 implantation sites, with 5 early and 3 late resorptions.

Dead fetuses:

effects observed, non-treatment-related

Description (incidence and severity):

In Female No. 1602755, there is 16 implantation sites with 1 dead foetus.
In Female No. 1602730, there is 17 implantation sites, with 9 dead foetus.

Changes in pregnancy duration:

no effects observed

Description (incidence and severity):

On D20 of pregnancy (D20pc), all mated females were necropsied and all foetuses were examined.

Changes in number of pregnant:

no effects observed

Other effects:

not examined

Effect levels (maternal animals)

Key result

Remarks on result:

not determinable due to absence of adverse toxic effects

Maternal abnormalities

Key result

Abnormalities:

no effects observed

Results (fetuses)

Fetal body weight changes:

no effects observed

Description (incidence and severity):

The mean body weight per litter and per group of live foetuses were indicated as well as the mean caudal-cranial measurement per litter and per group and the mean weight of the placenta of these foetuses per litter and per group.

Reduction in number of live offspring:

no effects observed

Changes in sex ratio:

no effects observed

Changes in litter size and weights:

no effects observed

Changes in postnatal survival:

not specified

External malformations:

effects observed, non-treatment-related

Description (incidence and severity):

There were isolated macroscopic findings seen in the control or treated groups at the same incidence such as point or area in head, limbs or back

Skeletal malformations:

no effects observed

Description (incidence and severity):

The test item administered in pregnant rats from Day 6 to Day 19 of gestation did not induce any relevant changes in foetuses examined at skeletal examination.

Visceral malformations:

no effects observed

Description (incidence and severity):

The test item administered in pregnant rats from Day 6 to Day 19 of gestation did not induce any relevant changes in foetuses examined at visceral examination.

Details on embryotoxic / teratogenic effects:

There was no change in pre and post-implantation loss, in the number of corpora lutea, in the number of live and abnormal foetuses, in the number of normal and abnormal dead foetuses or in the early and late resorptions.
There was no change in caudo-cranial measurement,foetus weight or proportion of male/female foetus.
There was a statistically significant lower placenta weight in the group receiving test item at 100 mg/kg.
This was low in amplitude (-6%) and was not attributed to a toxicological effect of test item.

Effect levels (fetuses)

Key result

Sex:

male/female

Remarks on result:

not determinable due to absence of adverse toxic effects

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 7.8.2/2: Clinical signs – autonomic profile/miscellaneous - Group incidences

Clinicalsigns	Time	Vehicle	Testitem100mg/kg	Testitem300mg/kg	Testitem1000mg/kg
Increasedsalivation	D5 D12 D13 D14 D15 D16 D17 D18 D19	0/20 0/20 0/20 0/20 0/20 0/20 0/20 0/20 0/20	0/20 0/20 0/20 0/20 0/20 0/20 0/20 0/20 0/20	0/20 0/20 1/20 1/20 0/20 0/20 0/20 0/20 0/19	0/20 1/20 0/20 2/20 1/20 1/19 3/19 2/19 2/19
Chromodacryorrhoea	D5 D9 D10 D11 D12 D14 D15 D17 D18 D19	0/20 0/20 0/20 0/20 0/20 0/20 1/20 0/20 0/20 0/20	0/20 0/20 0/20 0/20 0/20 0/20 0/20 0/20 0/20 0/20	0/20 1/20 1/20 1/20 1/20 0/20 0/20 1/20 1/20 0/19	0/20 0/20 1/20 0/20 0/20 1/20 2/20 1/19 1/19 1/19

Only times with clinical signs and showing differences with between groups are reported.

Results are expressed as the group incidence of animals showing the sign.

Corn oil

D: Day

P>0.05, when compared with the control group dosed with the vehicle, Fisher's test.

†: mortality occurred, no statistical analysis was performed.

Table 7.8.2/3: Body weight (mean table)

Treatment

D5

D6

D7

D8

D9

D10

D11

Vehicle

Mean SEM % N

260 6 NA 20

264 6 +2 20

267 6 +3 20

272 6 +5 20

277 6 +7 20

284 6 +9 20

288 6 +11 20

Testitem 100mg/kg

Mean SEM % N P

262 5 NA 20 NS

267 6 +2 20 NS

270 6 +3 20 NS

275 6 +5 20 NS

279 5 +6 20 NS

286 6 +9 20 NS

289 6 +10 20 NS

Testitem 300mg/kg

Mean SEM % N P

257 5 NA 20 NS

260 5 +1 20 NS

263 4 +2 20 NS

268 4 +4 20 NS

273 4 +6 20 NS

278 5 +8 20 NS

284 4 +11 20 NS

Testitem 1000mg/kg

Mean SEM % N P

262 5 NA 20 NS

266 5 +2 20 NS

268 4 +2 20 NS

273 4 +4 20 NS

277 4 +6 20 NS

283 5 +8 20 NS

288 4 +10 20 NS

Threshold

18

18

18

18

18

18

18

Treatment

D12

D13

D14

D15

D16

D17

D18

D19

D20

Vehicle

Mean SEM % N

295 6 +13 20

300 6 +15 20

306 6 +18 20

313 6 +20 20

323 6 +24 20

337 6 +30 20

350 6 +35 20

363 6 +40 20

376 6 +45 20

Testitem 100mg/kg

Mean SEM % N P

294 6 +12 20 NS

301 6 +15 20 NS

306 6 +17 20 NS

314 6 +20 20 NS

324 7 +24 20 NS

337 8 +29 20 NS

348 9 +33 20 NS

357 9 +36 20 NS

368 10 +40 20 NS

Testitem 300mg/kg

Mean SEM % N P

289 5 +12 20 NS

294 4 +14 20 NS

300 4 +17 20 NS

308 4 +20 20 NS

318 4 +24 20 NS

333 4 +30 20 NS

342 5 +33 20 NS

355 6 +38 19 NS

367 6 +43 19 NS

Testitem 1000mg/kg

Mean SEM % N P

293 4 +12 20 NS

299 4 +14 20 NS

306 5 +17 20 NS

311 5 +19 20 NS

320 5 +22 20 NS

336 5 +28 19 NS

348 5 +33 19 NS

359 5 +37 19 NS

370 6 +41 19 NS

Threshold

18

18

18

18

18

20

20

22

22

Results expressed in g

D: day

Vehicle: Corn oil

NS:P>0.05, when compared to control group

Analysis of variance for repeated measurements with Dunnett's test

NA: not applicable

%: variation expressed in percentage in relation to predose values

Table 7.8.2/4: Absolute weight of uterus (mean values)

Treatment		UterusWeight(g)
Vehicle	Mean SEM N	71.6 2.5 20
Testitem 100mg/kg	Mean SEM N % P	70.4 5.5 20 -2 NS
Testitem 300mg/kg	Mean SEM N % P	67.6 4.0 20 -6 NS
Testitem 1000mg/kg	Mean SEM N % P	69.6 3.9 20 -3 NS
	Threshold	14.0

%: variation expressed in percentage in relation to control values

Vehicle: Corn oil

NS:P>0.05, when compared with control group

Analysis of variance with Dunnett's test if P <0.05

Table 7.8.2/5: Macroscopic observations - Group incidences

Organs	Observations	vehicle	Testitem100mg/kg	Testitem300mg/kg	Testitem1000mg/kg
Placenta	Dark Total animals involved	0/256 0	0/259 0	14/243 14	0/246 0
Head	Punctate or point Area Total animals involved	1/256 2/256 3	0/259 2/259 2	1/243 1/243 2	1/246 2/246 3
Limbs	Punctate or point Area Total animals involved	1/256 2/256 3	2/259 3/259 5	1/243 5/243 6	1/246 4/246 5
Abdomen	Punctate or point Total animals involved	1/256 1	0/259 0	0/243 0	0/246 0
Tail	Area Twisted Total animals involved	1/256 0/256 1	0/259 0/259 0	0/243 1/243 1	0/246 0/246 0
Back	Punctate or point Area Total animals involved	14/256 0/256 14	10/259 5/259 15	11/243 0/243 11	11/246 2/246 13

Table 7.8.2/6: Caudo-cranial measurements and weights of live foetuses, weights of the placenta (mean values)

Treatment		Caudo-cranial measurement(mm)	Foetus weight(g)	Placenta weight(g)
Vehicle	Mean SEM N	35.2 0.2 256	3.77 0.03 256	0.567 0.006 255
Testitem 100mg/kg	Mean SEM N % P	34.8 0.1 259 -1 NS	3.69 0.02 259 -2 NS	0.533 0.005 259 -6 ??
Testitem 300mg/kg	Mean SEM N % P	34.7 0.3 243 -1 NS	3.70 0.04 243 -2 NS	0.557 0.006 242 -2 NS
Testitem 1000mg/kg	Mean SEM N % P	35.2 0.1 246 0 NS	3.78 0.02 246 0 NS	0.569 0.005 246 0 NS
	Threshold	0.6	0.09	0.018

%: variation expressed in percentage in relation to control values

Vehicle: Corn oil

NS:P>0.05, ??:P<0.01, when compared with control group

Analysis of variance with Dunnett's test if P <0.05

Applicant's summary and conclusion

Conclusions:

Under the experimental conditions adopted, N-(2-hydroxypropyl) Oleamide (Batch No. 160602013073 w/o Solvent) at 100, 300 and 1000 mg/kg administered daily by the oral route to the pregnant Sprague Dawley rats from day 6 to day 19 of gestation did not induce any changes on the embryo-foetal development.
The No Observable Adverse Effect Level (NOAEL) for embryo-foetal development was considered to be 1000 mg/kg.

Executive summary:

In a GLP-compliant prenatal developmental toxicity study performed according to OECD guideline 414, N-(2-hydroxypropyl) Oleamide diluted in corn oil was administered by gavage to groups of mated female Sprague-Dawley rats (20 mated females/dose) at the dose levels of 0, 100, 300, 1000 mg /kg bw/ day from Days 6 to 19 after mating.

The aim of the study was to investigate any possible adverse effects of N-(2-hydroxypropyl) Oleamide on the pregnant female rat and on the developing embryo and foetus following daily administration to pregnant rats by the oral route from implantion throughout organogenesis and until late gestation.

Morbidity/mortality checks were performed twice daily. Clinical observations were performed before the first dosing and daily.

Body weight was recorded on D5 then daily from D6 to D20. Food consumption was measured daily from D6 to D19.

On D20 of pregnancy (D20pc), all mated females were necropsied and all foetuses were examined.

Foetuses were examined macroscopically and the following data were noted:

• Number of live or dead foetuses

• Individual foetal body weight (live and dead)

• Caudo-cranial measurement of live and dead foetuses

• Gross evaluation and weight of placenta of all foetuses

• External morphological examination of these foetuses

• Sexing of all foetuses

The clinical signs (increased salivation and chromodacryorrhoea) observed were at low incidence and were not attributed to a toxicological effect of the test item.

There was no change in pre and post-implantation loss, in the number of corpora lutea, in the number of live and abnormal foetuses, in the number of normal and abnormal dead foetuses or in the early and late resorptions.

There was no change in caudo-cranial measurement, foetus weight or proportion of male/female foetus. There was a statistically significant lower placenta weight in the group receiving test item at 100 mg/kg. This was low in amplitude and was not attributed to a toxicological effect of test item.

The test item administered in pregnant rats from Day 6 to Day 19 of gestation did not induce any relevant changes in foetuses examined at skeletal and visceral examination.

Based on the above results, it can be concluded that the test item has no harmful effects on the prenatal development of the rat offspring at doses used in the present study.

Under the experimental conditions adopted, N-(2-hydroxypropyl) Oleamide (Batch No. 160602013073 w/o Solvent) at 100, 300 and 1000 mg/kg bw/day administered daily by the oral route to the pregnant Sprague Dawley rats from day 6 to day 19 of gestation did not induce any changes on the embryo-foetal development. The No Observable Adverse Effect Level (NOAEL) for embryo-foetal development was considered to be 1000 mg/kg bw/day.