Insulin Aspart Ethyl Ester

Genetic toxicity: in vivo

Administrative data

Endpoint:

in vivo mammalian somatic cell study: cytogenicity / erythrocyte micronucleus

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study without detailed documentation

Justification for type of information:

The present study, NDA report No. T-25, study nr. 940372, is described in a summary study report on Insulin aspart is based on GLP guideline studies prepared by Novo Nordisk. The summarised studies were performed as part of the non-clinical toxicity test regime for authorisation of Insulin aspart as human medicine and the studies are therefore in compliance with the guidelines for authorisation of human medicine.

Data source

Reference

Reference Type:

other company data

Title:

Unnamed

Year:

2001

Report date:

2001

Materials and methods

Test guideline

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 474 (Mammalian Erythrocyte Micronucleus Test)

GLP compliance:

yes

Type of assay:

other: micronuclei

Test material

Test material information

Constituent 1

Reference substance name:

Insulin aspart

Molecular formula:

C256H381N65O79S6

IUPAC Name:

Insulin aspart

Test material form:

solid: particulate/powder

Details on test material:

Molecular weight: 5793.6 Da

Specific details on test material used for the study:

Study performed using the active pharmaceutical ingredient Insulin aspart as test substance

Test animals

Species:

mouse

Strain:

not specified

Details on species / strain selection:

Not specified

Sex:

male/female

Details on test animals or test system and environmental conditions:

Not specified

Administration / exposure

Route of administration:

subcutaneous

Vehicle:

Not specified

Details on exposure:

Administration of 200 U/ml on two occasions, 10 hours apart, at a dose volume of 5 ml/kg, allowed a total dose of 2000 U/kg/day to be administered. This corresponds to approximately 2000 times the therapeutic dose, and was selected as the maximum dose administered. In addtion two lower dose levels of 1000 and 500 U/kg/day were administered.

Duration of treatment / exposure:

Two subcutaneous injection, 10 hours apart, over 1 day.

Frequency of treatment:

Ones (two subcutaneous injection, 10 hours apart)

Post exposure period:

Animals were sacrificed 24 or 48 hours after the second treatment.

Doses / concentrations

Doses / concentrations 1

Dose / conc.:

500 other: U/kg/day

Remarks:

Administered over two injections

Doses / concentrations 2

Dose / conc.:

1 000 other: U/kg/day

Remarks:

Administered over two injections

Doses / concentrations 3

Dose / conc.:

2 000 other: U/kg/day

Remarks:

Administered over two injections

No. of animals per sex per dose:

Not specified

Control animals:

yes, concurrent vehicle

Positive control(s):

Yes

Examinations

Tissues and cell types examined:

Polychromatic erythrocytes (PCE) from bone marrow

Details of tissue and slide preparation:

Not specified

Evaluation criteria:

Micronucleus frequency was estimate din 2000 PCE per animal

Statistics:

Not specified

Results and discussion

Test results

Key result

Sex:

male/female

Genotoxicity:

negative

Vehicle controls validity:

valid

Positive controls validity:

valid

Additional information on results:

Slides from all dose groups were analysed and micronucleus frequency estimated in 2000 PCE per animal. All positive control animals exhibited increased numbers of micronucleated PCE such that the micronucleus frequency in the positive control group was significantly greater than in concurrent controls.
Negative (vehicle) control mice exhibited normal group mean ratios of PCE to NCE and normal frequencies of micronucleated PCE within historical negative control ranges. Mice treated with insulin aspart at all doses exhibited group mean ratios of PCE to NCE and frequencies of micronucelated PCE which were similar to the values for vehicle control groups a both sampling times. There were no instances of statistically significant increases in micornucleus frequency for any of the groups receiving the test article at either sampling time.
The maximum dose caused a marked drop in blood glucose levels in treated mice.

Applicant's summary and conclusion

Conclusions:

Insulin aspart did not induce micronuclei in the polychromatic erythrocytes of the bone marrow of mice treated with subcutaneous injection using dose levels up to 2000 U/kg/day of inuslin aspart.

Executive summary:

Insulin aspart was teset for its ability to induce micronuclei in the bone marrow of subcutaneously dosed male and female mice. Administration of 2000, 1000 or 500 U/kg/day (divided in two injections) were administered to the mice. Postive and negative (vehicle) control animals were also included. All animals were sacrificed 24 or 48 hours after the second injection.

Slides from all dose groups were analysed and micronucleus frequency estimated in 2000 PCE per animal. All positive control animals exhibited increased numbers of micronucleated PCE such that the micronucleus frequency in the positive control group was significantly greater than in concurrent controls.

Negative (vehicle) control mice exhibited normal group mean ratios of PCE to NCE and normal frequencies of micronucleated PCE within historical negative control ranges. Mice treated with insulin aspart at all doses exhibited group mean ratios of PCE to NCE and frequencies of micronucelated PCE which were similar to the values for vehicle control groups a both sampling times. There were no instances of statistically significant increases in micornucleus frequency for any of the groups receiving the test article at either sampling time.

It was concluded that insulin aspart did not induce micronuclei in the polychromatic erythrocytes of the bone marrow of mice treated up to 2000 U/kg/day.