4-(methylamino)pentane-2-ol dibenzoate

Specific investigations: other studies

Administrative data

Endpoint:

endocrine system modulation

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

single dose application, 46.5 h exposure time

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

comparable to guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2014

Report date:

2014

Materials and methods

Test guideline

Qualifier:

equivalent or similar to guideline

Guideline:

other: OECD 455

Version / remarks:

The YES/YAS assay works with genetically modified yeast cells (human estrogen and androgen receptors hERa nd hAR, reporter gene lac-Z for ß-galactosidase on plasmid)

Deviations:

not applicable

Remarks:

1. experiment invalid due to calculation error

Principles of method if other than guideline:

The YES/YAS assay uses genetically modified yeast cells, which contain the human estrone and androgen receptors hERa and hAR. The receptor gene lac-Z, which encodes enzyme ß-galactosidas is also integrated in the cells.
If a test item interacts with a receptor, the reporte gene will be expressed and ß-galactosidase will be secreted into the medium. The medium contains an indicator (CPRG), which is converted by ß-galactosidase into a red product. This red product can be measured photometrically.

GLP compliance:

yes (incl. QA statement)

Remarks:

certified by Landesamt für Umwelt, Wasserwirtschaft und Gewerbeaufsicht Rheinland-Pfalz, Kaiser-Friedrich-Str. 7, D-55116 Mainz, Germany

Type of method:

in vitro

Test material

Test material information

Constituent 1

Reference substance name:

4-(methylamino)pentane-2-ol dibenzoate

EC Number:

807-124-1

Cas Number:

1519029-23-2

Molecular formula:

C20H23NO3

IUPAC Name:

4-(methylamino)pentane-2-ol dibenzoate

Test material form:

solid: particulate/powder

Specific details on test material used for the study:

4-(methylamino)pentan-2-ol dibenzoate, CAS No.: 15 19029-23-2, monoconstituent, solid, homogenous, stable at room temperature, soluble in DMSO
batch no. STCB/226/14, production date: Jun. 2014, purity: 96.06%

Test animals

Species:

other: yeast (Saccharomyces cerevisiae), genetically modified

Strain:

other: yeast, genetically modified: hERa and hAR receptors, plasmid containing lac-Z

Sex:

not specified

Details on test animals or test system and environmental conditions:

The YES/YAS Assay Kit was including Saccharomyces cerevisiae was obtained from Xenometrix AG, Gewerbestraße 25, CH-4123 Allschwil, Switzerland
batch no: N05-2785 (invalid experiment) and N05-2862 (valid experiment)

Administration / exposure

Route of administration:

other: dropped into culture flasks

Vehicle:

DMSO

Details on exposure:

After checking the yeast cultures for growth witha icroplate reader by measuring the optical density, 8 dilutions of positive controls and test item dilution respectively were pipetted into a 96-well dilution plate.
Next 4 assay plates were labelled (YES agonist/YAS agibost/YES antagonist/YAS antagonist) and filled with the corresponding medium. Then each positive control dilution and test item dilution was pipetted from the dilution plate to an assay plate in duplicate. After addition of yeast cells the plates were incubated at 32 +/-°C in a humidity container with agitation. Colour change of the medium in the positive controls indicated reaction after 46.5h.

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

Yeast cells were checked for growth by measuring of the optical density at 690 nm.

Duration of treatment / exposure:

46.5 h

Frequency of treatment:

single dose

Doses / concentrations

Doses / concentrations 1

Dose / conc.:

0 other: mol/L

Doses / concentrations 2

Dose / conc.:

0 other: mol/L

Doses / concentrations 3

Dose / conc.:

0 other: mol/L

Doses / concentrations 4

Dose / conc.:

0 other: mol/L

Doses / concentrations 5

Dose / conc.:

0 other: mol/L

Doses / concentrations 6

Dose / conc.:

0 other: mol/L

Doses / concentrations 7

Dose / conc.:

0 other: mol/L

Doses / concentrations 8

Dose / conc.:

0 other: mol/L

Details on study design:

Two experiments were performed. The first experiment was invalid, because the concentration of the stock solution of the test item was too low. Therefore the experiment was repeated. The second experiment was valid.
8 dilutions of positive controls or test item were examined in duplicate. Estrogen and androgen agonistic activities were detected directly by ß-galactodsidase activity. Antagonistic activities were detected by inhibition of ß-galactosidase activity in the presence of estrogen or androgen.
The solvent DMSO served as negative control. Cytotoxicity was checked be measurement of the cell density.

Examinations

Examinations:

Colour change by ß-galctosidase activity.
The ß-galactosidase activity correlates directly with the activating (agonists) or inhibiting (antagonists) activities of the test item.

Positive control:

YES agonist assay: 17ß-estradiol (E2)
YES antagonist assay: 4-hydroxytamoxifen (HT)
YAS agonist assay: 5a-dihyrotestosterone (DHT)
YAS antagonist assay: Flutamide (FL)

Results and discussion

Details on results:

Under the conditions of this test the test item 4-(methylamino)pentan-2-ol dibenzoate showed no estrogenic and androgenic agonistic or antagonistic properties.

Applicant's summary and conclusion

Conclusions:

Under the conditions of this YES/YAS assay no estrogenic or androgenic agonistic or antagonistic activities for the test item 4-(methylamino)pentane-2-ol dibenzoate were found.