Insulin Aspart Precursor

Genetic toxicity: in vivo

Administrative data

Endpoint:

in vivo mammalian cell study: DNA damage and/or repair

Remarks:

unscheduled DNA synthesis

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-26, study nr. 940374, 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

GLP compliance:

yes

Type of assay:

unscheduled DNA synthesis

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:

rat

Strain:

other: CD rats

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:

Insulin aspart was administered subcutaneously on two occasions approximately 10 hours apart at a dose volume of 5 ml/kg

Duration of treatment / exposure:

Two injectiions subcutaneusly, approximately 10 hours apart at a dose volume of 5 ml/kg

Frequency of treatment:

Two injections

Post exposure period:

2-4 hours after the second injection (12-14 hours after initial dosing), animals were killed and their liver perfused with collagenase to provide a primary culture of hepatocytes.

Doses / concentrations

Doses / concentrations 1

Dose / conc.:

2 000 other: U/kg/day

Remarks:

Administered over two doses of 1000 U/kg

Doses / concentrations 2

Dose / conc.:

800 other: U/kg/day

Remarks:

Administered over two doses

No. of animals per sex per dose:

5

Control animals:

yes, concurrent vehicle

Positive control(s):

yes

Examinations

Tissues and cell types examined:

hepatocytes from perfused livers

Details of tissue and slide preparation:

Livers were perfused with collagenase to provide primary cultures of hepatocytes. Cultures were made from three animals in each dose group and were treated with [3H] thymidine. Six slides from each animal were prepared with fixed hepatocytes and of these, three were dipped in photographic emulsion to prepare autoradiograms. The net grain count (NNG), the number of grains present in the nucleus minus the mean number of grains in three equivalent areas of cytoplasm, was determined for each of two of the three slides, for each animal and dose group.

Evaluation criteria:

Not specified

Statistics:

Not specified

Results and discussion

Test results

Key result

Sex:

male/female

Genotoxicity:

negative

Toxicity:

not specified

Vehicle controls validity:

valid

Positive controls validity:

valid

Additional information on results:

N/A

Any other information on results incl. tables

Negative (vehicle) control animals gave a group mean NNG ( net grain count) value of less than zero with only 0.7 -1.7% cells in repair. Group mean NNG values were increased by positive control treatment to at least 7.5%, with more than 50% cells found to be in repair. The vehicle control NNG value was consistent with both published and historical control data, and the system was shown to be sensitive to a known DNA damaging agent requiring metabolism for its action. The assay was therefore accepted as valid.

Treatment of male and female rats with 800 or 2000 U/kg/day insulin aspart subcutaneously, did not produce a group mean NNG value greater than -1.2, nor were any more than 1% cells found in repair at either dose level.

Applicant's summary and conclusion

Conclusions:

Insulin aspart did not to induce unscheduled DNA synsthesis at doses up to 2000 U/kg/day

Executive summary:

Treatment of male and female CD rats with 800 or 2000 U/kg/day insulin aspart subcutaneously, did not produce a group mean NNG value greater than -1.2, nor were any more than 1% cells found in repair at either dose level.

It was therefore concluded that insulin aspart failed to induce unscheduled DNA synsthesis detectable under the experimental conditions employed