2-Hydroxypropyl-β-cyclodextrine ethers

Repeated dose toxicity: oral

Administrative data

Endpoint:

sub-chronic toxicity: oral

Type of information:

experimental study

Adequacy of study:

supporting study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

study well documented, meets generally accepted scientific principles, acceptable for assessment

Data source

Reference

Reference Type:

publication

Title:

Unnamed

Year:

2010

Materials and methods

Test guideline

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 408 (Repeated Dose 90-Day Oral Toxicity Study in Rodents)

GLP compliance:

yes

Test material

Test material information

Constituent 1

Reference substance name:

.beta.-Cyclodextrin, 2-hydroxypropyl cycloheptaamylose

IUPAC Name:

.beta.-Cyclodextrin, 2-hydroxypropyl cycloheptaamylose

Constituent 2

Reference substance name:

-

EC Number:

420-920-1

EC Name:

-

Cas Number:

128446-35-5

Molecular formula:

Hill formula: (C42H70-nO35)(C3H7O)n; n(mittel)=5,25

IUPAC Name:

5,10,15,25-tetrakis(hydroxymethyl)-40,44,47,49-tetrakis(2-hydroxypropoxy)-20,30,35-tris[(2-hydroxypropoxy)methyl]-2,4,7,9,12,14,17,19,22,24,27,29,32,34-tetradecaoxaoctacyclo[31.2.2.2³,⁶.2⁸,¹¹.2¹³,¹⁶.2¹⁸,²¹.2²³,²⁶.2²⁸,³¹]nonatetracontane-36,37,38,39,41,42,43,45,46,48-decol

Test material form:

solid: particulate/powder

Remarks:

migrated information: powder

Details on test material:

Active ingredient: hydroxypropylated .beta.-cyclodextrin
% Active: 86.3
Nature and quantity of impurities: Propylene glycol, water, Na
Physical form: powder
Colour: white
Density: Not Applicable
Solubility: water soluble
pH: 6.8
Adsorptive properties (on glass, plastic): N/A
Volatility: non-volatile
Expiry date: 1 April 2002
Other relevant characteristics: N/A

Specific details on test material used for the study:

HPbCD was obtained from Cargill, Inc. (Cedar Rapids, IA)

Test animals

Species:

rat

Strain:

Sprague-Dawley

Details on species / strain selection:

Sprague-Dawley (Crl:CD[SD]VAF/Plus)

Details on test animals or test system and environmental conditions:

Sprague-Dawley (Crl:CD[SD]VAF/Plus) rats (10/sex/group) were obtained from Charles River Laboratories (Wilmington, MA). Rats were 6 ± 1 weeks old at initiation of dosing. Males were 220.7– 295.5 g and females 150.0–201.5 g at the initiation of dosing. All rats on study were individually housed in suspended, stainless steel cages, were maintained on a 12-h light cycle (temperature 72°F ± 4°F, 30–70% humidity), and were given water and certified rodent lab diet (PMI Nutrition International, Inc.) ad libitum.

Administration / exposure

Route of administration:

oral: gavage

Details on oral exposure:

5 ml/kg, calculated from most recent body weight
syringe and a ball-tipped gavage tube.

Analytical verification of doses or concentrations:

not specified

Duration of treatment / exposure:

92-93 days

Frequency of treatment:

daily

Doses / concentrations

Doses / concentrations 1

Dose / conc.:

500 mg/kg bw/day (nominal)

Doses / concentrations 2

Dose / conc.:

1 000 mg/kg bw/day (nominal)

No. of animals per sex per dose:

10/sex/dose

Control animals:

other: historical control

Examinations

Observations and examinations performed and frequency:

General observations: Body weight was measured weekly beginning 1 week prior to dosing, and a fasted terminal body weight was recorded prior to necropsy. Food consumption was measured weekly beginning 1 week prior to dosing. Rats were checked at least once daily for viability and clinical observations were recorded twice daily, at the time of dosing, and 1–3 h postdose.

Ophthalmologic examinations: Ophthalmologic examinations consisted of focal illumination and indirect ophthalmoscopy in a subdued light setting. Examinations were preformed 2 weeks prior to dosing and during weeks 5 and 11.

Hematology and serum chemistry evaluations: Blood for hematology and serum chemistry was collected during weeks 2 and 13. Total blood volume collected per interval for hematology and serum chemistry was 1 ml from mice, 1.5 ml from rats, and 3 ml from dogs and monkeys. Hematology analysis was performed using Cell- Dyn 3500 (Abbott Laboratories, Abbott Park, IL) and Sysmex R-3000 (Medical Electronics, Miami, FL) hematology analyzers. Serum chemistry parameters were assessed using a CX5CE/Delta (Beckman Instruments, Brea, CA).
Hematology parameters examined (measured and calculated): Erythrocyte count, Hemoglobin, Hematocrit, Mean corpuscular volume, Mean corpuscular hemoglobin, Mean corpuscular hemoglobin concentration, Reticulocyte count, Platelet count, Mean platelet volume, Total leukocyte count, Differential leukocyte, count Blood smear morphology. Serum chemistry parameters examined (measured and calculated) Hemolysis, Lipemia, Icterus, Glucose, Blood, urea, nitrogen, ALT, Aspartate aminotransferase, AP, Total bilirubin, Total protein, Albumin, Globulin, Albumin/globulin, ratio Creatinine GGT, Cholesterol, Triglycerides, Sodium, Potassium, Chloride, Calcium, Inorganic phosphorus.
Coagulation assessment: Blood for coagulation assessment was collected at necropsy. Prothrombin time and activated partial thromboplastin time were assessed using an STA Compact hemostasis analyzer (Diagnostica Stago, Inc., Parsippany, NJ).

Urinalysis: Freshly voided (4 h) and 24-h urine samples were collected from during weeks 2 and 13. Urine was collected within 24 h prior to blood collection for hematology and serum chemistry. Urinalysis parameters from the freshly voided (4 h) samples were evaluated using a CX5CE/Delta (Beckman Instruments), whereas urinalysis parameters from the 24-h samples were analyzed using a Bayer Clintek ATLAS (Bayer Diagnostics, Tarrytown, NY).
Parameters examined:
4-h Urinalysis/urine parameters examined—3-month: Color Clarity pH, Protein Glucose, Ketones, Bilirubin Blood Urobilinogen.
24-h Urinalysis/urine parameters examined—3-month: Volume, Sodium Chloride, Phosphorus, Osmolality, Potassium, Calcium (total), Creatinine clearance.

Sacrifice and pathology:

Macroscopic and histopathological evaluation: Rats were humanely euthanized by exsanguination under isoflorane anesthesia. All tissues were fixed in 10% neutral buffered formalin with the exception of the eyes (3% glutaraldehyde), harderian glands (3% glutaraldehyde), and testes (10% neutral buffered formalin in mice and rats; modified Davidson’s fixative in dogs and monkeys). Tissues were then trimmed, embedded in paraffin wax, and sectioned at 4–5 microns. All tissues were stained with hematoxylin and eosin, with the exception of the testes and epididymides, which were stained with periodic acid- Schiff stain. All tissues from all dose groups were examined by a boardcertified veterinary pathologist. Bone marrow smears were obtained from the sternum and were evaluated for cytology by a board-certified veterinary clinical pathologist.
Organweights measured: Ventral prostate gland, Liver, Spleen, Brain, Lungs, Thymus, Epididymides, Ovaries, Heart, Testes, Kidneys, Uterus (plus cervix).
Tissues collected and examined histopathologically: adrenal gland, Aorta—thoracic, Bone (Femur and sternum), Bone marrow (sternum), Brain, Epididymides,Esophagus, Eyes, Gall, bladder, Harderian glands, Heart, Kidneys, Large intestines (cecum and colon), Larynx/pharynx, Liver, Lungs, Lymph nodes (mandibular and mesenteric), Mammary gland, Ovaries, Pancreas, Parathyroid gland, Peripheral nerve (sciatic), Pituitary gland, Prostate gland, Salivary gland, Seminal vesicles, Skeletal muscle (biceps), Skin, Small intestines (duodenum, jejunum, ileum), Spinal cord (thoracolumbar), Spleen, Stomach, Testes, Thymus, Thyroid gland, Tongue, Trachea, Urinary bladder, Uterus (plus cervix),Vagina.

Statistics:

Statistics: Descriptive statistics (mean, index of variability, SEM, SD) were calculated for all parameters except ECGs. A student’s t-test was used to assess statistical significance of the serum transanimase values in mice and rats.

Results and discussion

Results of examinations

Clinical signs:

no effects observed

Mortality:

no mortality observed

Body weight and weight changes:

no effects observed

Food consumption and compound intake (if feeding study):

no effects observed

Food efficiency:

not specified

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

not specified

Ophthalmological findings:

no effects observed

Haematological findings:

no effects observed

Clinical biochemistry findings:

effects observed, treatment-related

Description (incidence and severity):

In female rats, treatment with 500 or 1000 mg/kg HPbCD resulted in time- and dose-dependent increases in serum AST and ALT levels. After 3 weeks of treatment with 1000 mg/kg HPbCD, serum AST and ALT levels were elevated (þ 38.2% and þ 31.1%, respectively), whereas after 13 weeks of treatment, serum AST and ALT levels were elevated in animals treated with 500 and 1000 mg/kg HPbCD (AST: þ 49.3% and þ 11.5%, ALT: þ 64.8% and þ 195.2%, respectively). In the 1000 mg/kg dose group at week 13, increases in serum AST and ALT levels were significantly higher than those seen at week 2, suggesting a progression of hepatic toxicity in these animals. AP and GGT were unchanged, and male rats were unaffected.

Urinalysis findings:

no effects observed

Behaviour (functional findings):

no effects observed

Immunological findings:

no effects observed

Organ weight findings including organ / body weight ratios:

no effects observed

Gross pathological findings:

no effects observed

Neuropathological findings:

not examined

Histopathological findings: non-neoplastic:

no effects observed

Description (incidence and severity):

There was no hepatic histopathologic correlate for the elevated transaminase findings, and liver weight and macroscopic appearance were normal. AST and ALT levels in male rats were similar to controls are all time points examined.

Histopathological findings: neoplastic:

no effects observed

Effect levels

1

Key result

Dose descriptor:

NOAEL

Effect level:

>= 1 000 mg/kg bw/day (nominal)

Based on:

test mat.

Sex:

male/female

Remarks on result:

not determinable due to absence of adverse toxic effects

2

Dose descriptor:

LOEL

Effect level:

500 mg/kg bw/day (nominal)

Based on:

test mat.

Sex:

female

Basis for effect level:

clinical biochemistry

Target system / organ toxicity

Key result

Critical effects observed:

no

Applicant's summary and conclusion

Conclusions:

In rats HP-beta-CD did not indcue adverse effects in rats when dosed orally for 90 days. The effect on liver enzymes is considered of an adaptive nature as neither liver weight nor histopathological examination showed a correlating effect. The NOAEL is considered >= 1000 mg/kg b.w.