Cycloheptapentylose

Developmental toxicity / teratogenicity

Administrative data

Endpoint:

developmental toxicity

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: OECD Guideline study report

Data source

Reference

Reference 1

Reference Type:

study report

Title:

Unnamed

Year:

1991

Report date:

1991

Reference 2

Reference Type:

publication

Title:

THE REPRODUCTIVE AND DEVELOPMENTAL TOXICITY PROFILE OF BETA·CYCLODEXTRIN IN RODENTS

Author:

PAUL C. BARROW, PHILlPPE OUVIER and DANIEL MARZlN

Year:

1995

Bibliographic source:

Reproductiuve Toxicology, 9 (4), 389- 398, 1995

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 414 (Prenatal Developmental Toxicity Study)

Deviations:

no

GLP compliance:

yes

Limit test:

no

Test material

Test material information

Constituent 1

Reference substance name:

Cycloheptapentylose

EC Number:

231-493-2

EC Name:

Cycloheptapentylose

Cas Number:

7585-39-9

Molecular formula:

C42H70O35

IUPAC Name:

5,10,15,20,25,30,35-heptakis(hydroxymethyl)-2,4,7,9,12,14,17,19,22,24,27,29,32,34-tetradecaoxaoctacyclo[31.2.2.2~3,6~.2~8,11~.2~13,16~.2~18,21~.2~23,26~.2~28,31~]nonatetracontane-36,37,38,39,40,41,42,43,44,45,46,47,48,49-tetradecol (non-preferred name)

Details on test material:

Name Beta-cyclodextrin (KLEPTOSE )
Origin Sample department
Packaging Cardboard sack
lot number: lot 0152. sample 422223.
Quantity required 10 kg
Date of receipt 10-Jan-90
Form White powder
The water content of Beta-cyclodextrin used in this study is 9.6 %) .

Test animals

Species:

rat

Strain:

Sprague-Dawley

Details on test animals or test system and environmental conditions:

Animal species: rat
Strain: OFA originating from Sprague-Dawley
Source: IFFA CREDO (Les Oncin. France)
Sex: male and female
Age: 12 weeks
Weight: 257 - 288 grams
Number: sufficient number of virgin females in order to obtain 20 pregnant fe-males/group.
Rationale for species selection: Species usually used for this type of trial, and recom-mended by the OECD Guideline 414
Animal husbandry Housing: Makrolon cages, stainless-steel grid lid size 37.5 x 23.5 x 16 cm bedding of chopped maize cobs 5 animals/cage
Food: Pelleted diet UAR base 210 from Usine d'Alimentation Rafionnelle, Villemoisson-sur-Orge, France
Drinking water: Polypropylene water-bottles, Tap water, autoclaved for 10 minutes at 120 °C, changed 3 times a week.
Food and water were available at libitum. The quantities consumed were measured 3 times a week..
Environment: Temperature: 20 +/- 2 °C, humidity 50 +/- 10%, artificial lighting 12 hours/day, air changes 12 times/hour
The animals were housed under quarantine conditions for at least one week prior to the commencement of the study.

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

CMC (carboxymethyl cellulose)

Details on exposure:

The beta-Cyclodextrin suspensions were prepared daily at concentrations of 1.25, 2.5 and 5g/10ml of 1% CMC. They were stored at + 4°C after use and kept at the disposal of the laboratory for testing of the concentration. They were discarded after one week.

VEHICLE
- Justification for use and choice of vehicle (if other than water): solubility
- Concentration in vehicle: The test substance was suspended in carboxymethyl cellulose (CMC). This slurry was prepared by mixing 10 g CMC in 1 1 of water for injection (1%). and administered by oral gavage.
The doses are 0 ; 1.25 ; 2. 5 and 5 g/kg/d by gastric intubation in a volume of 10 ml/kg. These doses correspond to those ingested during a subchronic toxicity study in the rat.

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

B-cyclodextrin levels in the suspension were measured by High Performance Liquid Chromatography (HPLC).
The brand of the equipment used was WATERS (MILLIPORE - WATERS. USA) and comprised:an isocratic pump (type WATERS 510),a sampler (type WATERS 710B) or any injection system (type Rheodyne) with a loop of 10 microlitres, a differential refractometer (type WATERS R410), a recorder-integrator (type Shimadzu CRSA).The analytical column was SUPERCOSIL LC-18 5 microns (SUPELCO ref 5-B985).

Details on mating procedure:

Each female was cage with a male on day 0, from the end of the afternoon until the following morning. A cervical smear was performed the day after mating. The presence of spermatozoa) detected .by microscopic examination. reveals the coitus had occured. The female was thus considered mated and at day 1 of gestation. The gestation is confirmed on day 14 by abdominal palpation .

Duration of treatment / exposure:

from day 7 to day 16 the period of organogenesis

Frequency of treatment:

once daily

Duration of test:

On day 21 of gestation, the females were sacrificed (ether) and subjected to a post-mortem examination .

No. of animals per sex per dose:

20 pregnant females

Control animals:

yes, concurrent vehicle

Details on study design:

- Dose selection rationale: These doses correspond to those ingested during a subchronic toxicity study in the rat. Thus the highest dose was considered as close to the No Observable Effect Level .
- Rationale for animal assignment (if not random): random

Examinations

Maternal examinations:

CAGE SIDE OBSERVATIONS: Yes
- Time schedule: Daily
- Cage side observations checked in table were included.


DETAILED CLINICAL OBSERVATIONS: no


BODY WEIGHT: Yes
- Time schedule for examinations: days 1, 3, daily from 7 to 16, 21


FOOD CONSUMPTION : Yes

WATER CONSUMPTION: Yes

POST-MORTEM EXAMINATIONS: Yes
- Sacrifice on gestation day 21

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

Fetal examinations:

- External examinations: Yes: [all per litter ]
- Soft tissue examinations: Yes: [half per litter ]
- Skeletal examinations: Yes: [half per litter ]
- Head examinations: Yes: [ half per litter ]

Statistics:

The following statistical methods were performed by means of Statgraphics and SAS programs.
1. Preliminary
From the data collected, three other variables were calculated: By Litter
VAR 13 - Percentage of resorptions over the number of implants
VAR 14 - Percentage of males over the number of live foetuses
VAR 15 - Ratio number af females/number of males.
2. Descriptive statistics:
The same calculations were done for each of the four groups as follows
• For the variables concerning the dams or litters (var. 1 to 9, 13, 14, 15)
• Firstly, calculation of some caracteristics (mean, minimum, maximum, standard devia-tion. standard error) on all dams of the group.
• On the other hand, only for var. 4 to 9. determination of the distribution of all dams ac-cording to the various values of the variable studied.
• For the variables concerning the foetuses (var. 10, 11, 12) :
• First, for each litter, calculation of the mean (and standard deviation) body weight. size. and determination of the distribution of the sex on all foetuses of the litter.
• On the other hand. on all dams of the group. calculation of the caracteristics previously cited and determination of the distribution of sex .
3. Groups comparison :
A univariate comparison (one variable at a time) between the 4 groups was first done; then, if a significant difference was observed. a multivariate comparison was performed. The tests used depend upon the type of variable analysed. The different tests used are : • One-factor variance analysis on the raw data or after arc sin Vp transformation on the percentage.
• Nested two~factor analysis of variance and n repetitions on raw values.
• Nested three-factor analysis of variance and n repetitions on raw values. For each variance analysis. if a significant difference is observed. then group homogeneity is de-termined by the test of Ouncan, Scheffe and LSD.
• Non-parametric Kruskall and Wallis test
• Chi-square test comparing several distributions observed

Indices:

no data

Historical control data:

given in report

Results and discussion

Results: maternal animals

Maternal developmental toxicity

Details on maternal toxic effects:

Maternal toxic effects:yes

Details on maternal toxic effects:
At the high dose level only, a statistically significant reduction of body weight gain in females, accompanied by a reduction of food consumption.

Effect levels (maternal animals)

1

Dose descriptor:

NOAEL

Effect level:

2 500 mg/kg bw/day (actual dose received)

Basis for effect level:

other: maternal toxicity

2

Dose descriptor:

LOAEL

Effect level:

5 000 mg/kg bw/day

Basis for effect level:

other: maternal toxicity

3

Dose descriptor:

NOAEL

Effect level:

5 000 mg/kg bw/day

Basis for effect level:

other: developmental toxicity

Results (fetuses)

Details on embryotoxic / teratogenic effects:

Embryotoxic / teratogenic effects:no effects

Details on embryotoxic / teratogenic effects:
not applicable

Effect levels (fetuses)

Dose descriptor:

NOAEL

Effect level:

5 000 mg/kg bw/day

Basis for effect level:

other: teratogenicity

Fetal abnormalities

Abnormalities:

not specified

Overall developmental toxicity

Developmental effects observed:

not specified

Applicant's summary and conclusion

Conclusions:

Substance induced slight reduction of body weight gain in the high dose group but no embryotoxic or fetotoxic effects in any of the tested groups.

Executive summary:

A teratogenicity study of .beta.-cyclodextrin (92.5 % pure) according to OECD 414 was carried out in Sprague-Dawley rats. Four groups, each comprising 20 females, beta Cyclodextrin was administered by oral gavage at levels of 0; 1250; 2500 and 5000 mg/kg body weight/d respectively from day 7 to day 16. The day the presence of spermatozoa was observed on the vaginal smear was considered as day 1 of gestation and the females were sacrificed on day 21 of gestation.

In the high dose group five females died from gavage accidents between day 7 and day 17. Necropsy revealed oesophageal perforation for the 5 females. On day 1, there were no statistically significant differences between control and each treated group concerning the body weight variability or mean. The treated animals presented a reduced body weight gain from the beginning of the treatment period. The difference was statistically significant for the high dose group from day 8. At cessation of treatment, the body weight gain of the animals of this group was similar to that of the controls but their body weight at day 21 was still significantly decreased

During the first and third weeks of the study there were no significant . differences between control ·and treated groups concerning food consumption. During the second week which corresponds to the treatment period, a significant decrease in food consumption was· noted for mid and high dose groups and ( - 7.4 and - 19,2 %).

No statistical differences were noted on food conversion efficiency during the 3 weeks, between control and treated groups, despite an increase observed during week 2 for mid and high dose groups. 

Water consumption was similar in all groups during the first week.

The visceral examination by's technique and skeletal examination by alizarin technique of rat fetuses. from darns treated during the sensitive phase of embryogenesis with the test substance at the oral doses of 1250, 2500 and 5000 mg/kg/day did not reveal any detectable embryotoxic or teratogenic effect.

Following parameters were examined in litters: weight of full and empty uterus, weight of placenta, number of live and dead foetuses, number of implantation sites and resorptions, number of corpora lutea, ratio - resorption/implantation. No differences are noted between the groups.

The following parameters were examined in foetuses: weight and size of the foetuses, sex, ratio male/live foetuses, ratio female/male. The only significant difference in both sexes is the increased size of the foetuses in groups low and mid dose groups, compared with controls: +0 0.3 (0.84%) and + 0.4 mm (1.14%) respectively for males and females in low dose group ; + 0.6 (1.68%) and + 0.6 mrn (1.72%) respectively for males and females in the mid dose group. 

During the study, it was observed that the administration of beta-cyclodextrin at doses of 0, 1250, 2500 or 5000 mg/kg induced at the high dose level only, a statistically significant reduction of body .weight gain in females, accompanied by a reduction of food consumption. On the other hand, these females showed difficulty in swallowing the gavage suspension, and five of them died following oesophageal perforation. The dose level of 5000 mg/kg in a volume of 10 ml/kg is the limit of ingestion in this type of study . The lower dose of 2500 mg/kg per day had no toxic effect on the females. However. this produced no observable differences in the weight of the uteri and placentae, the number of live (100%) or dead (0%) foetuses, the number of implantation sites, resorptions and corpora lutea The same applies to the following foetal parameters: size, body weight, sex ratio. visceral and skeletal examinations. which indicate no embryotoxic or teratogenic effects ascribable to the treatment.

No embryotoxic or teratogenic effects were observed. It is concluded that beta-cyclodextrin administration at levels up to 2500 mg/kg bw/d had no adverse effect observable on pregnant females and at levels up to 5000 mg/kg bw/d had no adverse effect observable in that kind of study on litters and foetuses.