N-(4-chlorobenzhydryl)piperazine

Endpoint summary

Key value for chemical safety assessment

Effects on fertility

Description of key information

Toxicity to reproduction:

Waiver as the study dose not to be conducted because a pre-natal developmental toxicity study is avaliable.

Link to relevant study records

Reference

Endpoint:

screening for reproductive / developmental toxicity

Data waiving:

study scientifically not necessary / other information available

Justification for data waiving:

the study does not need to be conducted because a pre-natal developmental toxicity study is available

Effect on fertility: via oral route

Endpoint conclusion:

no study available

Quality of whole database:

Waiver

Effect on fertility: via inhalation route

Endpoint conclusion:

no study available

Effect on fertility: via dermal route

Endpoint conclusion:

no study available

Effects on developmental toxicity

Description of key information

Devlopmental toxicity study, NOAEL was considered  to be 12.5 mg/kg/day  as no developmental effects was observed and LOAEL was considered to be 25 mg/ kg/bw on the bases of  malformations, i.e., cleft palate with glossopalatine fusion, brachygnathia, micromelia, microstomia and inhibition of calcification  was observed at given dose level. When  female Sprague-Dawley rats were exposed  with  Norchlorcyclizine (303-26-4) through the 12-15 day of gestation by oral gavage. based on the above studies on Norchlorcyclizine(303 -26 -4) and its structurally similar read across substance chlorcyclizine hydrochloride (14362-31-3),It was considered that it had teratogenic potential on the bases of skeleton malformation observed in both the rodent species i.e rats and mice when exposed 11-14 days through gestation by oral gavage and also supported by study done using Intrauterine application .

Thus, comparing this value with the criteria of CLP regulation Norchlorcyclizine(303 -26 -4) can be classified as “Category 2”for reproductive toxicity.

Link to relevant study records

Reference

Endpoint:

developmental toxicity

Type of information:

experimental study

Adequacy of study:

supporting study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

data from handbook or collection of data

Justification for type of information:

Data from peer reviewed journal

Qualifier:

equivalent or similar to guideline

Guideline:

other: As mentioned below

Principles of method if other than guideline:

To evaluate the teratogenic potential of Norchlorcyclizine in rats.

GLP compliance:

not specified

Limit test:

no

Specific details on test material used for the study:

- Name of the test material: 1-[(4-chlorophenyl)(phenyl)methyl]piperazine
- IUPAC name: 1-[(4-chlorophenyl)(phenyl)methyl]piperazine
- Molecular weight: 286.804 g/mol
- Molecular formula: C17H19ClN2
- Substance type: Organic
- Smiles: c1([C@@H](c2ccccc2)N2CCNCC2)ccc(Cl)cc1
- Inchi: 1S/C17H19ClN2/c18-16-8-6-15(7-9-16)17(14-4-2-1-3-5-14)20-12-10-19-11-13-20/h1-9,17,19H,10-13H2

Species:

rat

Strain:

Sprague-Dawley

Details on test animals or test system and environmental conditions:

No data available

Route of administration:

oral: gavage

Vehicle:

water

Details on exposure:

PREPARATION OF DOSING SOLUTIONS:
Test material diluted with water
DIET PREPARATION
- Rate of preparation of diet (frequency):
- Mixing appropriate amounts with (Type of food):
- Storage temperature of food:

VEHICLE
- Justification for use and choice of vehicle (if other than water):
- Concentration in vehicle: 0,20,40,60,80 and 100 mg/kg
- Amount of vehicle (if gavage): 1.0ml
- Lot/batch no. (if required):
- Purity:

Analytical verification of doses or concentrations:

not specified

Details on mating procedure:

- Impregnation procedure: [artificial insemination / purchased timed pregnant / cohoused]
- If cohoused:
- M/F ratio per cage: No data available
- Length of cohabitation: No data available
- After ... days of unsuccessful pairing replacement of first male by another male with proven fertility.No data available
- Further matings after two unsuccessful attempts: [no / yes (explain)]No data available
- Verification of same strain and source of both sexes: [yes / no (explain)]No data available
- Proof of pregnancy: [vaginal plug / sperm in vaginal smear] referred to as [day 0 / day 1] of pregnancy:The day of finding sperm is now called day 1 of pregnancy rather than day 0.
- Any other deviations from standard protocol:No data available

Duration of treatment / exposure:

3 days (Through gestation day 13-16)

Frequency of treatment:

Drug was administered, in general, on days 13-16 rather than on previously designated days 12-15.

Duration of test:

19 days

Remarks:

0,20,40,60,80 and 100 mg/kg

No. of animals per sex per dose:

No data available

Control animals:

yes, concurrent vehicle

Details on study design:

No data available

Maternal examinations:

No data available

Ovaries and uterine content:

No data available

Fetal examinations:

- External examinations: Yes:
- Soft tissue examinations: No data
- Skeletal examinations: Yes: (At day 21 of gestation the palate was observed for the presence of a cleft by simply opening the mouth. At days 17,18, and 19, frozen fetal heads were first sectioned with a razor blade so as to remove the mandible; the tongue was then removed. The palate
could then be observed with the aid of low magnification, either directly or after brief staining with mildly alkaline 0.5 % aqueous alizarin red S.)
- Head examinations: Yes:

Statistics:

Yes statistics was observed.

Indices:

No data available

Historical control data:

No data available

Clinical signs:

not specified

Dermal irritation (if dermal study):

not specified

Mortality:

not specified

Body weight and weight changes:

not specified

Food consumption and compound intake (if feeding study):

not specified

Food efficiency:

not specified

Water consumption and compound intake (if drinking water study):

not specified

Ophthalmological findings:

not specified

Haematological findings:

not specified

Clinical biochemistry findings:

not specified

Urinalysis findings:

not specified

Behaviour (functional findings):

not specified

Immunological findings:

not specified

Organ weight findings including organ / body weight ratios:

not specified

Gross pathological findings:

not specified

Neuropathological findings:

not specified

Histopathological findings: non-neoplastic:

not specified

Histopathological findings: neoplastic:

not specified

Other effects:

not specified

Number of abortions:

not specified

Pre- and post-implantation loss:

not specified

Total litter losses by resorption:

not specified

Early or late resorptions:

not specified

Dead fetuses:

not specified

Changes in pregnancy duration:

not specified

Description (incidence and severity):

Migrated Data from removed field(s)
Field "Effects on pregnancy duration" (Path: ENDPOINT_STUDY_RECORD.DevelopmentalToxicityTeratogenicity.ResultsAndDiscussion.ResultsMaternalAnimals.MaternalDevelopmentalToxicity.EffectsOnPregnancyDuration): not specified

Changes in number of pregnant:

not specified

Other effects:

not specified

Dose descriptor:

other: not specified

Based on:

test mat.

Basis for effect level:

other: not specified

Remarks on result:

other: not specified

Abnormalities:

not specified

Localisation:

not specified

Fetal body weight changes:

not specified

Description (incidence and severity):

Migrated Data from removed field(s)
Field "Fetal/pup body weight changes" (Path: ENDPOINT_STUDY_RECORD.DevelopmentalToxicityTeratogenicity.ResultsAndDiscussion.ResultsFetuses.FetalPupBodyWeightChanges): not specified

Reduction in number of live offspring:

not specified

Changes in sex ratio:

not specified

Changes in litter size and weights:

not specified

Changes in postnatal survival:

not specified

External malformations:

not specified

Skeletal malformations:

effects observed, treatment-related

Description (incidence and severity):

Statistically significant Cleft palate was observed at 30 mg/kg/day dose level.

Visceral malformations:

not specified

Other effects:

not specified

Dose descriptor:

LOAEL

Effect level:

30 mg/kg bw/day (nominal)

Based on:

test mat.

Sex:

not specified

Basis for effect level:

skeletal malformations

Remarks on result:

other: Cleft palate was observed.

Abnormalities:

effects observed, treatment-related

Localisation:

other: Cleft palate was observed.

Developmental effects observed:

yes

Lowest effective dose / conc.:

30 mg/kg bw/day (nominal)

Treatment related:

yes

Relation to maternal toxicity:

not specified

Dose response relationship:

not specified

Relevant for humans:

not specified

Conclusions:

Developmental toxicity study, LOAEL was considered to be 30 mg/kg/day for Norchlorcyclizine (303-26-4) in male and female Sprague Dawley rats fetuses when there dams where exposed for 13-16 days through gestation by oral gavage.

Executive summary:

Developmental toxicity study was observed in male and female Sprague Dawley rats fetuses when there dams were exposed for 13-16 days through gestation by oral gavage for Norchlorcyclizine .The test material diluted with water .The day of finding sperm is now called day 1 of pregnancy rather than day 0and the pregnant female Sprague Dawley rats  were exposed through the 13-16 days of gestation at concentration of 0, 20,40,60,80 and 100 mg/kg. At day 21 of gestation the palate was observed for the presence of a cleft by simply opening the mouth. At days 17,18, and 19, frozen fetal heads were first sectioned with a razor blade so as to remove the mandible; the tongue was then removed. The palate could then be observed with the aid of low magnification, either directly or after brief staining with mildly alkaline 0.5 % aqueous alizarin red S. Statistically significant Cleft palate was observed at 30 mg/kg/day in fetuses’ .Therefore LOAEL was considered to be 30 mg/kg/day for Norchlorcyclizine (303-26-4) in male and female Sprague Dawley rats fetuses when there dams were exposed for 13-16 days through gestation by oral gavage.

Effect on developmental toxicity: via oral route

Endpoint conclusion:

adverse effect observed

Dose descriptor:

LOAEL

25 mg/kg bw/day

Study duration:

subacute

Species:

rat

Quality of whole database:

Data is Klimicsh 2 and from peer reviewed journal

Effect on developmental toxicity: via inhalation route

Endpoint conclusion:

no study available

Effect on developmental toxicity: via dermal route

Endpoint conclusion:

no study available

Additional information

Developmental toxicity

In different studies, Norchlorcyclizine(303 -26 -4) has been investigated for developmental toxicity to a greater or lesser extent. Often are the studies based on in vivo experiments and estimated data in rodents, i.e. most commonly in rats for Norchlorcyclizine(303 -26 -4) also been compared with the experimental studies performed on structurally similar read across substance Tartrazine(1934-21-0).

An experimental study conducted by C. T. G. KING, S. A. WEAVER AND S. A. NARROD (Journal of Pharmacology and Experimental Therapeutics. 147,391, 1965) Developmental toxicity study for Norchlorcyclizine(303 -26 -4) in male and female Sprague-Dawley rats when there dams were exposed through the 12-15 day of gestation by oral gavage as a metabolite of antihistamines was observed .They were exposed at different concentration  12.5,25, 37.5,50 and 125 mg/kg/day. No significant change was observed at the resorption rate and the number of mean nidation sites at any dose level in dams. Fetal death at a dose level of 125 mg/kg /day was observed. Norchlorcyclizine induces malformations, i.e., cleft palate with glossopalatine fusion, brachygnathia, micromelia, microstomia and inhibition of calcification at dose level 25 mg/kg body weight /day. Therefore NOAEL was considered to be 12.5 mg/kg/day as no developmental effects was observed and LOAEL was considered to be 25 mg/ kg/bw on the bases of Norchlorcyclizine induces malformations, i.e., cleft palate with glossopalatine fusion, brachygnathia, micromelia, microstomia and inhibition of calcification at given dose level. When female Sprague-Dawley rats were exposed with Norchlorcyclizine (303-26-4) through the 12-15 day of gestation by oral gavage.

It is supported by experimental study conducted by HERBERT S. POSNER AND ALZENA DARR (TOXICOLOGY AND APPLIED PHARMACOLOGY 17,67-75(1970)) Developmental toxicity study was observed in male and female Sprague Dawley rats fetuses when there dams were exposed for 13-16 days through gestation by oral gavage for Norchlorcyclizine .The test material diluted with water .The day of finding sperm is now called day 1 of pregnancy rather than day 0and the pregnant female Sprague Dawley rats  were exposed through the 13-16 days of gestation at concentration of 0, 20,40,60,80 and 100 mg/kg. At day 21 of gestation the palate was observed for the presence of a cleft by simply opening the mouth. At days 17,18, and 19, frozen fetal heads were first sectioned with a razor blade so as to remove the mandible; the tongue was then removed. The palate could then be observed with the aid of low magnification, either directly or after brief staining with mildly alkaline 0.5 % aqueous alizarin red S. Statistically significant Cleft palate was observed at 30 mg/kg/day in fetuses’ .Therefore LOAEL was considered to be 30 mg/kg/day for Norchlorcyclizine (303-26-4) in male and female Sprague Dawley rats fetuses when there dams were exposed for 13-16 days through gestation by oral gavage.

It is further supported by experimental study conducted by C. T. G. KING JOYCE HOWELL (American J of Obst. & Gynec, 1996 Vol 95 Is 1 PP 109-111) on structurally similar read across substance chlorcyclizine hydrochloride (14362-31-3). Developmental toxicity study of chlorcyclizine hydrochloride (14362-31-3) was performed on Forty-one mature female NIH albino mice weighing 20 ±3 grams. The compound was intubated in water solution (not to exceed 0.5 ml.) from the eleventh to the fourteenth day of gestation in dose concentration150, 200,250,350mg/kg. Onset of gestation was established by the presence of a vaginal plug and the day following recorded as the first of pregnancy. 325 fetuses examined for malformations namely, cleft palate, fusion of the tongue to the palatine shelves, micrognathia, microstomia, micromelia, and incomplete calcification of the axial skeleton. Treatment with chlorcyclizine hydrochloride (350 mg. per kilogram) yielded malformations in 100 per cent of the young. At 250 mg. per kilogram the incidence was 72 per cent and at 200 mg. per kilogram 68.5 per cent. Administration of this compound at 150 mg. per kilogram did not induce malformations in this strain of mice. Furthermore, it is suggested that their teratogenic action is mediated by the formation of the demethylated metabolite, norchlorcyclizine, Hence NOAEL was considered to be 150 mg/kg/ as no malformation observed and LOAEL was considered to be 250mg/kg on the bases of malformation observed in 68% fetuses .When female mice were exposed with chlorcyclizine hydrochloride (14362-31-3) for 11-14 days through gestation by oral gavage.

 Thus, based on the above studies on Norchlorcyclizine(303 -26 -4) and its structurally similar read across substance chlorcyclizine hydrochloride (14362-31-3),It was considered that it had teratogenic potential on the bases of skeleton malformation observed in both the rodent species i.e rats and mice when exposed 11-14 days through gestation by oral gavage and also supported by study done using Intrauterine application .Thus, comparing this value with the criteria of CLP regulation Norchlorcyclizine(303 -26 -4) can be classified for developmental toxicity.

 

Toxicity to reproduction: other studies

Description of key information

 Developmental toxicity via other route: Intrauterine application

The teratologenic potential of Norchlorcyclizine (303-26-4) was evaluated by A. L. WILK(Teratology Vol 2, Is 1, PP 55-65, feb 1969) using Intrauterine application technique. The teratognicity of Norchlorcyclizine (303-26-4) was studied in female Sprague-Dawley rats when they were exposed through the 14-15 day of gestation after intrauterine application of test material. In this technique of intrauterine application a compound, which by passes the metabolism of the adult rat and directly subjects the fetus to microgram concentrations of the compound for a known period of time. Pregnant Sprague-Dawley rats were used on days 11-16 of gestation. Vaginal smears were taken to determine pregnancy with the appearance of sperm designated as day 0 of gestation. Semisterile operational procedure was followed. Animals weighing approximately 200 g were anesthetized with an intraperitoneal injection of1ml of a 10% pentobarbital sodium solution supplemented, when necessary, with light ether inhalation. A midline abdominal incision was made and the double horned uterus was exteriorized.Asmall cut was then made through the uterine wall and a Millipore filter square impregnated with norchlorcyclizine HCl, or HCl was inserted either on the yolk sac over the intact amniotic sac (fetal side) or on the placenta .Once inserted, the filters remained in place until after day 16 when fetal movements seemed to dislodge them. In each pregnant rat, fetuses in one uterine horn were usually subjected to experimental filters and the HCl-containing, control filter was placed over fetuses in the other horn. Prior to day 14 of gestation the position of the embryo cannot be seen through the uterus and the filters were always inserted centrally over the fetus. On days 14 and 15 it was possible to insert the filters centrally, over the hindlimb, or over the head area .All fetuses observed on days 14, 15, and 16 appeared to lie with their left side uppermost and the right side toward the placenta. After these uterine manipulations the uterus was returned to the abdominal cavity, the muscle layer sutured, and the skin clamped with stainless steel clips. The animal was then placed on its back in a warm recovery area and when it recovered from the anesthesia it was returned to the animal room and gestation was allowed to continue to day 20.Pregnant rats were weighed and killed on day 20 and the young removed. Gross malformations were noted by observation and the fetuses were weighed and then placed in 80% ethanol. After alcohol fixation the fetuses were cleared of most soft tissue with 1% KOH and stained with 0.5% alizarin redS.The stained specimens were examined for skeletal malformations. The effect of the surgical procedure on the maternal weight gain from days 13-20 and on the fetal weight on day 20. Postoperational survival of pregnant rats was excellent. Pregnant operated rats gained approximately one-third as much weight as did unoperated pregnant animals and operated fetuses had a mean weight of about 3.3g on day 20while that of unoperated fetuses was 3.9 g. Fetal survival after surgery and insertion of HCl-containing filters was over 60% on every day except day 14 when it was approximately 42%. Cleft palate was produced at rates of 29 and 35% after norchlorcyclizine (50ug/filter) was inserted centrally on the fetal side on day 13 or 14 respectively. When the concentration of norchlorcyclizine was increased to 90 or 100 ug/filter and implanted centrally or over the head area of fetuses on day 15, less than 3% cleft palate was observed. Another malformation frequently seen following intrauterine application of norchlorcyclizine, was a limb malformation, which affected either the left forelimb or left hindlimb depending on whether the filter was placed centrally or over the hindlimb area of the fetus. The severity of the malformation ranged from simple fusion of the digits to complete absence of a paw. Left forelimb anomalies occurred at a rate of 24 and 38% after norchlorcyclizine (50 ug/filter) was inserted centrally on day 13 or 14 respectively and at a rate of about 3% after norchlorcyclizine- filter insertion on day 11, 12, 15, or 16Apositive relation between filter position over the fetus and cleft palate production was also found .When norchlorcyclizine (50 ug/filter) was inserted over the head area on the fetal side 42% of the fetuses had cleft palates and 13% had hindlimb malformations. Filters with similar concentration of norchlorcyclizine inserted over the left hind limb, however, resulted in 11% cleft palate and 55% left hind limb malformation. Gross observation of whole and cleared fetuses showed that the heads of some fetuses treated with test material filters were misshapen and the bodies more squat than normal. The control filters produced no malformation when implanted on any of the experimental days. Similarly no abnormalities resulted after norchlorcyclizine-filter insertion over the placenta on day 13.Hence LOAEL was considered to be 0.05mg/kg/day on the bases of teratogenic potential. When female Sprague-Dawley rats were exposed with Norchlorcyclizine (303-26-4) through the 13-14 day of gestation after intrauterine application

 

Justification for classification or non-classification

Thus, comparing this value with the criteria of CLP regulation Norchlorcyclizine(303 -26 -4) can be classified as “Category 2”for reproductive toxicity.