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Toxicological information

Genetic toxicity: in vivo

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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, Novo Nordisk preclinical expert report No. T-13, study nr. LSR 85/NLP005/508, is described in a tabular summary study report on Human Insulin 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 Human Insulin 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

Constituent 1
Chemical structure
Reference substance name:
Human Insulin
Molecular formula:
C257H383N65O77S6
IUPAC Name:
Human Insulin
Test material form:
solid: particulate/powder
Remarks:
White powder
Details on test material:
Molecular formula: C257H383N65O77S6
Molecular weight: 5807.66 g/mol
Specific details on test material used for the study:
Study performed using the active pharmaceutical ingredient Human Insulin as test substance

Test animals

Species:
mouse
Strain:
not specified
Details on species / strain selection:
CD-1 Charles River BR Lab (UK)
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:
S.C. administration of 4000, 800, 160 U/kg as single dose in the morning.

Sampling time 24, 48 and 72 hours after treatment.
Number of animals analyzed per group: 5 males and 5 females (160-800 U/kg) and 15 males and 15 females (4000 U/kg)

Positive control: Chlorabucil (60mg/kg, single oral administration) analyzed 24 post exposure.

Negative control: Acrapid Medium.

Duration of treatment / exposure:
One subcutaneous injection in the morning.
Frequency of treatment:
Onces (one subcutaneous injection in the morning)
Post exposure period:
Animals were sacrificed 24, 48 and 72 hours after exposure.
Doses / concentrationsopen allclose all
Dose / conc.:
160 other: U/kg
Dose / conc.:
800 other: U/kg
Dose / conc.:
4 000 other: U/kg
No. of animals per sex per dose:
above 15 animals/group
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.
There were no instances of statistically significant increases in micornucleus frequency for any of the groups receiving the test article at either sampling time.

Applicant's summary and conclusion

Conclusions:
Human Insulin did not induce micronuclei in the polychromatic erythrocytes of the bone marrow of mice treated with subcutaneous injection using dose levels up to 4000 U/kg of Human inuslin.
Executive summary:

Human insulin was teset for its ability to induce micronuclei in the bone marrow of subcutaneously dosed male and female mice. Administration of 4000, 800 or 160 U/kg (one s.c. injection) were administered to the mice. Postive and negative (vehicle) control animals were also included. All animals were sacrificed 24, 48 or 72 hours after administration.

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.

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 4000 U/kg.