Method for identifying malodour counteractants

Patent metadata

Field	Value
Publication number	US-11448642-B2
Application number	US-201716468522-A
Country	US
Kind code	B2
Filing date	Dec 14, 2017
Priority date	Dec 14, 2016
Publication date	Sep 20, 2022
Grant date	Sep 20, 2022

Abstract

The invention relates to a method for determining whether a test compound can counteract malodour from a sulphur odorant. The method includes contacting the test compound and the sulphur odorant with an olfactory receptor OR4E2, and comparing the binding of the OR4E2 to the sulphur odorant, or the activity of the OR4E2, in the presence and in the absence of the test compound.

First claim

The invention claimed is:

1. A method for determining whether a test compound can counteract a perceived malodour from a sulfur odorant, the method comprising the steps of:
a) contacting cells expressing an OR4E2 polypeptide with the sulfur odorant in the presence and in the absence of the test compound;
b) measuring an activity of the OR4E2 polypeptide in the presence and in the absence of the test compound under an identical condition;
c) comparing the activity of the OR4E2 polypeptide in the presence of the test compound to the activity of the OR4E2 polypeptide in the absence of the test compound;
wherein a decrease in the activity of the OR4E2 polypeptide in the presence of the test compound relative to the activity of the OR4E2 polypeptide in the absence of the test compound identifies the test compound as a compound that can counteract the perceived malodour from the sulfur odorant;
wherein the sulfur odorant is selected from the group consisting of: a sulfanyl alkanol selected from the group consisting of 2-mercapto-3-methyl-1-butanol, 3-mercapto-2-methyl-1-pentanol, 3-mercapto-2-methyl-1-propanol, 3-mercapto-3-methyl-1-butanol, 3-mercapto-1-hexanol, 3-methyl-3(2-methyldisulfanyl)-butan-1-ol and 3-mercapto-3-methyl-1-hexanol; and a sulfide selected from the group consisting of methyl-2-propenyl disulfide, 2,4-dithiapentane, dimethyl sulfide, dimethyl disulfide, dimethyl trisulfide, diallyl sulfide, diallyl disulfide and diallyl trisulfide; and
wherein the activity of the OR4E2 polypeptide is measured using a reporting system capable of producing a readable or measurable signal.

2. The method of claim 1 , wherein said cells are mammalian cells.

3. The method of claim 2 , wherein said mammalian cells are selected from human embryonic kidney cells, Chinese hamster ovary cells and HeLa cells.

4. The method of claim 1 , wherein step a) is performed in the presence of Cu 2+ ions.

5. The method of claim 1 , wherein the OR4E2 polypeptide has the sequence of SEQ ID NO: 1.

6. The method of claim 1 , wherein the OR4E2 polypeptide is encoded by a nucleic acid having the sequence of SEQ ID NO: 2.

7. The method of claim 1 , wherein the reporting system comprises a gene whose transcription is driven by a cAMP response element which is stimulated by adenylate cyclase which is activated by a stimulatory G-protein.

8. A method for determining whether a test compound can counteract a perceived malodour from a sulfur odorant, the method comprising the steps of:
a) providing cells expressing an OR4E2 polypeptide;
b) measuring a magnitude of a response of the OR4E2 polypeptide to control conditions;
c) contacting said cells with the sulfur odorant in the presence and in the absence of the test compound under an identical condition;
d) measuring a magnitude of a response of the OR4E2 polypeptide to the sulfur odorant in the absence of the test compound under an identical condition;
e) calculating a first fold of change (FOC1), wherein FOC1 equals the magnitude of the response measured in step d) divided by the magnitude of the response measured in step b);
f) measuring a magnitude of a response of the OR4E2 polypeptide to the sulfur odorant in the presence of the test compound under an identical condition;
g) calculating a second fold of change (FOC2), wherein FOC2 equals the magnitude of the response measured in step f) divided by the magnitude of the response measured in step b);
h) comparing FOC1 with FOC2; and
i) identifying the test compound as a compound that can counteract the perceived malodour from the sulfur odorant if a ratio FOC2/FOC1 is less than 0.8;
wherein the sulfur odorant is selected from the group consisting of: a sulfanyl alkanol selected from the group consisting of 2-mercapto-3-methyl-1-butanol, 3-mercapto-2-methyl-1-pentanol, 3-mercapto-2-methyl-1-propanol, 3-mercapto-3-methyl-1-butanol, 3-mercapto-1-hexanol, 3-methyl-3(2-methyldisulfanyl)-butan-1-ol and 3-mercapto-3-methyl-1-hexanol; and a sulfide selected from the group consisting of methyl-2-propenyl disulfide, 2,4-dithiapentane, dimethyl sulfide, dimethyl disulfide, dimethyl trisulfide, diallyl sulfide, diallyl disulfide and diallyl trisulfide.

9. The method of claim 8 , wherein the test compound is identified as the compound that can counteract the perceived malodour from the sulfur odorant if the ratio FOC2/FOC1 is less than 0.7.

Assignees

• Takasago Perfumery Co Ltd

Inventors

• Warr Jonathan
• Winnig Marcel

Classifications

• G01N2333/726

  G protein coupled receptor, e.g. TSHR-thyrotropin-receptor, LH/hCG receptor, FSH · CPC title

• G01N33/566

  using specific carrier or receptor proteins as ligand binding reagents {where possible specific carrier or receptor proteins are classified with their target compounds} · CPC title

• G01N33/5008 Primary

  for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics · CPC title