Insulin Aspart Ethyl Ester

Genetic toxicity: in vitro

Administrative data

Endpoint:

in vitro cytogenicity / chromosome aberration study in mammalian cells

Type of information:

experimental study

Adequacy of study:

key study

Study period:

Not specified

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

comparable to guideline study with acceptable restrictions

Justification for type of information:

The present study, NDA report No. T-24, study nr. 940373, 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:

according to guideline

Guideline:

OECD Guideline 473 (In Vitro Mammalian Chromosome Aberration Test)

GLP compliance:

yes

Type of assay:

other: chromosome aberration

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:

The study was conducted with the active pharmaceutical ingredient Insulin Aspart

Method

Target gene:

not specified

Species / strain

Species / strain / cell type:

lymphocytes: human periferal blood lymphocytes

Details on mammalian cell type (if applicable):

human lymphocyte cell cultures from a male and female donor

Metabolic activation:

with and without

Metabolic activation system:

rat liver post-mitochondrial fraction (S9) from aroclor 1254 induced animals

Test concentrations with justification for top dose:

up to 5000 ug/ml.

Vehicle / solvent:

Not specified

Controls

Untreated negative controls:

yes

Remarks:

Histroical negative controls

Negative solvent / vehicle controls:

yes

True negative controls:

not specified

Positive controls:

not specified

Details on test system and experimental conditions:

Eksperiment 1:
Treatment in the absence of S9 was continuous for 20 hours.

Treatment in the presence of S9 was for 3 hours, followed by 17 hours recovery period prior to harvest.

The test item concentrations for chromosome analysis were selected by evalutating the effect of insulin aspart on mitotic index. Chromosome aberrtions were analysed at three consecutive treatments concentrations up to 5000 ug/ml.


Eksperiment 2:
Treatment in the absence of S9 was continuous for 20 hours.

Treatment in the presence of S9 was for 3 hours, followed by 17 hours recovery period prior to harvest.

A delayed sampling time compared to experiment one of 44 hours and pulse treatment in the absesence of S9 was included in this experiment.

Chromosome aberrations were analysed in celss receiving 20 hour continuous treatments in the absence of S9 and 3 hour pulse treatment in its presence at three consecutive treatment concentrations up to 5000 ug/ml.
The effect of a singe concentration only (without S9) were also investigated at the delayed (44 hour) sampling time after pulse and continuous treatment and following the 3 hour treatment in the absence of S9, at the 20 hour sampling time.

NUMBER OF REPLICATIONS:
2

Rationale for test conditions:

The test item concentrations for chromosome analysis were selected by evalutating the effect of insulin aspart on mitotic index. Chromosome aberrtions were analysed at three consecutive treatments concentrations up to 5000 ug/ml.

Evaluation criteria:

Comparison to historical negative controls and solvent controls.

Statistics:

N/A

Results and discussion

Test results

Key result

Species / strain:

lymphocytes: Human periferal blood lymphocytes

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

not specified

Vehicle controls validity:

not specified

Untreated negative controls validity:

not specified

Positive controls validity:

not specified

Additional information on results:

No data

Applicant's summary and conclusion

Conclusions:

Insulin aspart did not induce chromosome aberrations in the cultured human periferal blood lymphocytes when tested up to a concentration of 5000 ug/ml in either the absece or presence of S9

Executive summary:

The in vitro clastogenic potential of Insulin aspart was investigated in cultured human peripheral blood lymphocytes. Induction of chromosomal aberrations was investigated in two experiments at concentrations up to 5000 ug/ml with and without metabolic activation by S9. The treated lymphocytes had frequiencies of cells with aberrations which were similar to and not significantly different from those in concurrent solvent controls. Numbers of aberrant cells in all treated cultures fell within historical negative control ranges.

It was therefore concluded that Insulin aspart does not induce chromosome aberrations in cultured human periferal blood lymphocytes when tested up to a concentration of 5000 ug/ml in either the absece or presence of S9.