WO2018190118A1 - Method for screening perfume material having amber gris note - Google Patents

Abstract

Provided is a method for screening a perfume material having an amber gris note. The screening method according to the present invention, whereby a candidate compound having an amber gris note is screened from test compounds by using an olfactory receptor showing responsiveness to a perfume having an amber gris note, comprises: (i) a step for contacting the test compounds with an olfactory receptor selected from the group consisting of OR7A17, OR7C1 and a protein, said protein containing an amino acid sequence having at least 80% identity to the amino acid sequence represented by SEQ ID NO: 2 or 4 and showing responsiveness to a perfume having an amber gris note, and measuring the responses of the olfactory receptor to the test compounds; (ii) a step for measuring the response of the olfactory receptor used in step (i) in the absence of the test compounds; and (iii) a step for comparing the measurement results of steps (i) and (ii) and selecting a test compound which shows a change in the responsiveness as a candidate compound having an amber gris note.

Description

Screening method for perfume material presenting Ambergris notes

The present invention relates to a method for screening a candidate compound for a fragrance material exhibiting Ambergris notes.

Ambergris is an important fragrance along with musk in the scent of animatic fragrance in perfume. Ambergris is a natural fragrance obtained from sperm whale stones drifting to the coast, but it is very valuable due to the prohibition of commercial whaling and so on. is there. Each fragrance company is developing a fragrance that exhibits Ambergris Note with better performance.
On the other hand, in recent years, a method using cultured cells expressing olfactory receptors has been proposed in order to search for new fragrance material candidates (for example, Japanese Patent No. 5520153 (Patent Document 1)). Japanese Patent No. 5912226 (Patent Document 2).
In vertebrates, olfactory receptors are sensor proteins that are expressed mainly in the olfactory epithelium in the nasal cavity and respond to various aroma compounds. When the aroma compound binds to the olfactory receptor, the intracellular G protein is activated, and the G protein activates adenylate cyclase to convert ATP into cyclic AMP (cAMP). cAMP opens a CNG channel, whereby sodium ions (Na ⁺ ) and calcium ions (Ca ²⁺ ) flow into cells to generate receptor potentials and send information to the brain. It is considered possible to discriminate odors by measuring intracellular cAMP increase and cation increase when aroma compounds are brought into contact with cultured cells expressing olfactory receptors.
It is known that there are less than 400 types of human olfactory receptors, and one kind of odorant compound responds to one or more kinds of olfactory receptors, and one kind of olfactory receptor responds to several kinds of odorant compounds. Are known.

Japanese Patent No. 5520153 Japanese Patent No. 5912226

There is a possibility that a candidate compound exhibiting Ambergris notes can be screened by searching for an olfactory receptor that responds to a known aroma compound exhibiting Ambergris notes and using the olfactory receptor. However, olfactory receptors that respond to perfume materials that exhibit Ambergris notes have not been known so far.
Under such circumstances, it is desired to search for olfactory receptors that respond to a fragrance material exhibiting Ambergris notes and to provide a method for screening candidate compounds for new fragrance materials exhibiting Ambergris notes.

As a result of intensive studies to solve the above problems, the present inventor succeeded in identifying an olfactory receptor that specifically responds to AMBROXAN (registered trademark) or Amber Xtreme (trademark) representing Ambergris Note. did. The present inventor has further studied and found that by using these olfactory receptors, a candidate compound for a fragrance material exhibiting Ambergris notes can be appropriately screened, and the present invention has been completed.

 
で示される化合物からなる群より選択される、［１］に記載の方法。
［３］嗅覚受容体を発現している生体から単離された細胞上または嗅覚受容体を遺伝子操作により人為的に発現させた細胞上で嗅覚受容体の応答を測定する、［１］または［２］に記載の方法。
［４］嗅覚受容体の応答をレポーターアッセイにより測定する、［１］から［３］のいずれか一項に記載の方法。
［５］工程（ｉ）における測定結果を、工程（ｉｉ）における測定結果で割ることで求められるＦｏｌｄ　Ｉｎｃｒｅａｓｅ値が２以上であるときに、工程（ｉｉｉ）においてアンバーグリスノートを呈する候補化合物として選択する、［１］から［４］のいずれか一項に記載の方法。 That is, this invention provides the method of screening the candidate compound of the fragrance | flavor raw material which exhibits the Ambergris note shown below.
[1] A method of screening candidate compounds exhibiting Ambergris notes from test compounds using an olfactory receptor that exhibits responsiveness to a fragrance exhibiting Ambergris notes,
(I) A group comprising OR7A17, OR7C1, and a protein comprising an amino acid sequence having 80% or more identity with the amino acid sequence represented by SEQ ID NO: 2 or 4, and responsive to a fragrance exhibiting Ambergris notes Contacting a test compound with a more selected olfactory receptor and measuring a response of the olfactory receptor to the test compound;
(Ii) measuring the response of the olfactory receptor used in step (i) in the absence of the test compound;
(Iii) comparing the measurement results in steps (i) and (ii), and selecting a test compound with altered responsiveness as a candidate compound exhibiting Ambergris notes.
[2] The fragrance presenting Ambergris Note has the following formula:

The method according to [1], which is selected from the group consisting of compounds represented by:
[3] The response of the olfactory receptor is measured on a cell isolated from a living body expressing the olfactory receptor or on a cell in which the olfactory receptor is artificially expressed by genetic manipulation, [1] or [ 2].
[4] The method according to any one of [1] to [3], wherein the response of the olfactory receptor is measured by a reporter assay.
[5] When the Fold Increase value obtained by dividing the measurement result in step (i) by the measurement result in step (ii) is 2 or more, it is selected as a candidate compound that exhibits an Ambergris note in step (iii) The method according to any one of [1] to [4].

By using the method of the present invention, candidate compounds exhibiting Ambergris notes can be screened from test compounds. The screening method of the present invention is expected to contribute to the development of a new fragrance material that exhibits Ambergris notes. In addition, in order to develop a new fragrance material, evaluating the odors of many candidate compounds only by human olfaction may cause problems of olfactory fatigue and individual differences, and it may be difficult to select candidate compounds appropriately. However, according to the method of the present invention, such a problem can be solved or reduced.

It is a figure which shows the measurement result of the response of various olfactory receptors with respect to a compound (a). It is a figure which shows the measurement result of the response of the various olfactory receptors with respect to a compound (b). It is a figure which shows the measurement result of the response of olfactory receptor OR7A17 with respect to compound (a) of various density | concentrations. It is a figure which shows the measurement result of the response of olfactory receptor OR7C1 with respect to compound (b) of various density | concentrations. It is a figure which shows the measurement result of the response of olfactory receptor OR7A17 with respect to a compound (c). It is a figure which shows the measurement result of the response of olfactory receptor OR7C1 with respect to a compound (d). It is a figure which shows the measurement result of the response of olfactory receptor OR7C1 with respect to a compound (e). It is a figure which shows the measurement result of the response of olfactory receptor OR7C1 with respect to a compound (f). It is a figure which shows the measurement result of the response of olfactory receptor OR7C1 with respect to a compound (g). It is a figure which shows the measurement result of the response of olfactory receptor OR7A17 and OR7C1 with respect to various compounds.

Hereinafter, the screening method of the present invention will be specifically described.

The present invention is a method for screening a candidate compound exhibiting Ambergris notes among test compounds using an olfactory receptor that exhibits responsiveness to a fragrance exhibiting Ambergris notes,
(I) A group comprising OR7A17, OR7C1, and a protein comprising an amino acid sequence having 80% or more identity with the amino acid sequence represented by SEQ ID NO: 2 or 4, and responsive to a fragrance exhibiting Ambergris notes Contacting a test compound with a more selected olfactory receptor and measuring a response of the olfactory receptor to the test compound;
(Ii) measuring the response of the olfactory receptor used in step (i) in the absence of the test compound;
(Iii) comparing the measurement results in steps (i) and (ii), and selecting a test compound whose responsiveness has changed as a candidate compound exhibiting Ambergris notes.

The screening method of the present invention is responsive to a fragrance comprising OR7A17, OR7C1, and an amino acid sequence having 80% or more identity with the amino acid sequence represented by SEQ ID NO: 2 or 4, and exhibiting an Ambergris note Candidate compounds exhibiting Ambergris notes are selected from the test compounds using the responsiveness of the test compound to an olfactory receptor selected from the group consisting of proteins as an index.
Since one kind of olfactory receptor is known to respond to multiple kinds of aroma compounds having similar structures, an olfactory receptor that shows responsiveness to a known fragrance exhibiting Ambergris Note By evaluating the responsiveness of the test compound to the olfactory receptor, candidate compounds exhibiting Ambergris notes can be selected from the test compounds. In the present specification, the term “fragrance material exhibiting Ambergris note” refers to a compound, composition or mixture serving as a material for the flavor exhibiting Ambergris note. Hereinafter, each step of the screening method of the present invention will be described.

<Process (i)>
In step (i), a protein comprising OR7A17, OR7C1, and an amino acid sequence having 80% or more identity with the amino acid sequence represented by SEQ ID NO: 2 or 4, and responsive to a fragrance exhibiting Ambergris notes An olfactory receptor selected from the group consisting of is contacted with a test compound, and the response of the olfactory receptor to the test compound is measured.

The olfactory receptor includes OR7A17, OR7C1, and an amino acid sequence having an identity of 80% or more with the amino acid sequence represented by SEQ ID NO: 2 or 4, and is responsive to a fragrance exhibiting Ambergris notes An olfactory receptor selected from the group consisting of is used.
OR7A17 is a protein that is registered in GenBank as NM — 030901 and has an amino acid sequence (SEQ ID NO: 2) encoded by the first to 930th DNAs of the base sequence represented by SEQ ID NO: 1.
OR7C1 is registered in GenBank as NM_198944, and is a protein consisting of an amino acid sequence (SEQ ID NO: 4) encoded by the first to 963rd DNAs of the base sequence represented by SEQ ID NO: 3.
OR7A17 responds strongly to compound (a) and compound (c), and OR7C1 responds selectively and strongly to compound (b), compound (d), compound (e), compound (f), compound (g), etc. Therefore, the screening method using OR7A17 and OR7C1 is expected to contribute to the development of a fragrance material exhibiting Ambergris notes.

Amino acids having 80% or more, preferably 85% or more, more preferably 90% or more, still more preferably 95% or more, particularly preferably 98% or more identity with the amino acid sequence of these olfactory receptors as olfactory receptors An olfactory receptor selected from the group consisting of proteins comprising a sequence and responsive to a fragrance exhibiting Ambergris notes may be used. In this specification, the sequence identity of amino acid sequences is calculated by a BLAST search algorithm (available publicly from NCBI).

In the present invention, the fragrance exhibiting Ambergris note is preferably one or more selected from the group consisting of compounds represented by the following formula.

 

 

Compound (a) (3aR, 5aS, 9aS, 9bR) -3a, 6,6,9a-tetramethyl-2,4,5,5a, 7,8,9,9b-octahydro-1H-benzo [e] [1] Benzofuran is known by the name AMBROXAN (registered trademark) and is available from Kao Corporation.
Compound (b) (2,2,6,6,7,8,8-heptamethyl-3,3a, 4,5,5a, 7,8a, 8b-octahydrocyclopenta [g] [1] benzofuran) is obtained from Amber Xtreme. It is the main component of a fragrance known by the name of (trademark) and is available from International Flavors and Fragrances.
Compound (c) (1- (1,2,3,4,5,6,7,8-octahydro-2,3,8,8, -tetramethyl-2-naphthyl) ethane-1-one) is represented by ORBITONE It is known by the name (registered trademark) and is available from Takasago International Corporation.
The compound (d) (1R, 6S)-(1- (2,2,6-Trimethylcyclohexyl) hexan-3-ol) is known under the name Ambestol® or NORLIMBANOL® DEXTRO, It is available from Takasago International Corporation or Firmenich.
Compound (e) (2- (2,2,7,7-Tetramethyltriclo [6.2.1.0] (1,6) undec-5-en-5-yl) propan-1-ol and 2- ( 2,2,7,7-Tetramethyltriclo [6.2.1.0] (1,6) undec-4-en-5-yl) propan-1-ol isomer mixture) is AMBERMAX® And is available from Givaudan.
Compound (f) ((4aR, 5R, 7aS, 9R) -Octahydro-2,2,5,8,8,9a-hexylmethyl-4H-4a, 9-methanezuleno [5,6-d] -1,3- (dioxole) is known under the name AMBROCENEIDE ™ and is available from Symrise.
Compound (g) (3,8,8,11a-Tetramethyldodecahydro-5H-3,5a-epoxy-naphth (2,1-c) oxepin and isomers), known under the name AMBERKETAL, is obtained from Givaudan Is possible.

Among these, as the fragrance exhibiting Ambergris Note, the compound (a), the compound (b), the compound (c) and the compound (e) are more preferable, and the compound (a) and the compound (e) are particularly preferable. In the present specification, “showing responsiveness” means that some change is observed in olfactory receptor-expressing cells as compared to the absence of a fragrance.

Olfactory receptors may be used singly or in combination of two or more.

In the present invention, the method for contacting the olfactory receptor with the test compound and measuring the response of the olfactory receptor to the test compound is not particularly limited. For example, a test compound may be contacted on a cell isolated from a living body expressing the olfactory receptor and the response of the olfactory receptor may be measured, or the olfactory receptor may be artificially expressed by genetic manipulation. The olfactory receptor response may be measured by contacting with a test compound on the cultured cells. The time for which the olfactory receptor is brought into contact with the test compound depends on the concentration of the test compound and cannot be generally specified, but is usually 2 to 4 hours.

A cell in which an olfactory receptor is artificially expressed by genetic manipulation can be produced by transforming the cell with a vector incorporating a gene encoding the olfactory receptor.

In a preferred embodiment of the present invention, the N-terminal 20 amino acid residues of bovine rhodopsin may be incorporated together with the olfactory receptor. Incorporation of the N-terminal 20 amino acid residues of bovine rhodopsin can promote cell membrane expression of olfactory receptors.
Bovine rhodopsin is registered as NM_001014890 with GenBank. Bovine rhodopsin is a protein consisting of an amino acid sequence (SEQ ID NO: 6) encoded by the first to 1047th DNA of the base sequence shown in SEQ ID NO: 5.
Further, in place of bovine rhodopsin, identity with the amino acid sequence represented by SEQ ID NO: 6 is 80% or more, preferably 85% or more, more preferably 90% or more, still more preferably 95% or more, particularly preferably 98% or more. A protein that includes an amino acid sequence having the above and can promote cell membrane expression of an olfactory receptor may be used.
In addition, as long as it can promote the cell membrane expression of an olfactory receptor, not only bovine rhodopsin but also amino acid residues of other proteins may be used.

The method for measuring the response of the olfactory receptor is not particularly limited, and any method used in this field can be used. For example, when an aroma compound binds to an olfactory receptor, the intracellular G protein is activated, the G protein activates adenylate cyclase, and ATP is converted into cyclic AMP (cAMP). It is known to increase. Therefore, the response of the olfactory receptor can be measured by measuring the amount of cAMP. As a method for measuring the amount of cAMP, an ELISA method, a reporter gene assay method, or the like is used. Among them, it is preferable to measure the response of the olfactory receptor by a reporter gene assay method using a luminescent substance such as luciferase.

<Process (ii)>
In step (ii), the response of the olfactory receptor used in step (i) is measured in the absence of the test compound.
As a method for measuring the response of the olfactory receptor, a method similar to the method shown in step (i) can be used except that the olfactory receptor and the test compound are not brought into contact with each other. For example, the response of an olfactory receptor may be measured on a cell isolated from a living body expressing the olfactory receptor, or the olfactory receptor may be measured on a cell in which the olfactory receptor is artificially expressed by genetic manipulation. Body response may be measured. In order to appropriately compare the measurement results in steps (i) and (ii), the measurement conditions in steps (i) and (ii) are preferably the same except for the presence or absence of contact with the test compound.

<Process (iii)>
In step (iii), the measurement results in steps (i) and (ii) are compared, and the test compound whose responsiveness has changed is selected as a candidate compound that exhibits Ambergris notes.
In the present invention, when the measurement results in steps (i) and (ii) are compared and responsiveness changes, the test compound used in step (i) is evaluated as a candidate compound exhibiting Ambergris Note. be able to.
According to one embodiment of the present invention, the change in responsiveness may be evaluated using as an index the Fold Increase value obtained by dividing the measurement result in step (i) by the measurement result in step (ii). For example, when the response of an olfactory receptor is measured by a reporter gene assay using a luminescent substance such as luciferase, a test compound having a Fold Increase value of preferably 2 or more, more preferably 4 or more, and even more preferably 10 or more is used. It can be evaluated as a candidate compound exhibiting a grease note.

As described above, candidate compounds exhibiting Ambergris notes can be screened from test compounds. According to the present invention, candidate compounds exhibiting Ambergris notes can be selected from a large number of test compounds without causing problems due to olfactory fatigue or individual differences due to sensory evaluation performed by human olfaction.
The selected compound can be used as a candidate compound for a fragrance material exhibiting Ambergris notes. Based on the selected compound, it may be modified as necessary to develop a compound having an optimal odor. Furthermore, it is also possible to develop a fragrance material having an optimal odor by blending the selected compound with other fragrance materials. By using the screening method of the present invention, it is possible to contribute to the development of a new fragrance material exhibiting Ambergris notes.

Hereinafter, the present invention will be described in more detail with reference to examples, but the present invention is not limited to these examples.

[Example 1]
Search for olfactory receptors responding to perfume compound (a) and compound (b) exhibiting Ambergris notes (1) Olfactory receptor gene cloning Human olfactory receptors OR7A17 and OR7C1 genes are based on sequence information registered in GenBank. In addition, it was obtained by cloning by PCR from Human Genomic DNA: Female (Promega). The human olfactory receptor gene expression vector was obtained by incorporating the N-terminal 20 amino acid residues of bovine rhodopsin into the pME18S vector and further incorporating the obtained human olfactory receptor gene downstream thereof.

(2) Expression of olfactory receptor gene in HEK293T cells 0.05 μg of human olfactory receptor gene expression vector, 0.01 μg of RTP1S vector, 0.01 μg of firefly luciferase vector pGL4.29 (Promega) containing cAMP response element promoter, Then, 0.005 μg of Renilla luciferase vector pGL4.74 (Promega) containing thymidine kinase promoter was dissolved in 10 μL of Opti-MEM I (Thermo Fisher Scientific) to obtain a gene solution (for 1 hole). HEK293T cells are seeded in a 96-well plate (Biocoat, Corning) 100 μL at a cell count that reaches confluence after 24 hours, and a gene solution is added to each well by lipofection according to the usage of Lipofectamine 3000. After introduction, the cells were cultured at 37 ° C. in a 5% carbon dioxide atmosphere for 24 hours.

(3) Luciferase Reporter Gene Assay After removing the culture solution, 50 μL of each prepared in CD293 (Thermo Fisher Scientific) medium (20 μM L-glutamine added) so that the concentration of the fragrance compound as a sample is measured. After the addition and stimulation for 3 hours, luciferase activity was measured according to the method of use of Dual-Luciferase Reporter Assay System (Promega). The response intensity of the olfactory receptor was expressed as a fold increase value obtained by dividing the luciferase activity produced by stimulation of the fragrance compound by the luciferase activity produced in the test system not containing the fragrance compound.

(4) Measurement of olfactory receptor response to compound (a) and compound (b) In cells expressing OR7A17 and OR7C1, compound (a) (Kao Corporation) 100 μM and compound (b) (International Flavors and Fragrances, Inc.) ) Receptor response to 300 μM was measured by luciferase reporter gene assay. The results are shown in FIGS. 1A and B, respectively. Only OR7A17 responded to the compound (a), and only OR7C1 responded to the compound (b) with a Fold Increase value of 10 or more.
On the other hand, OR2L8 and OR4K5 described in the Mock test using cells that did not express the olfactory receptor and the response to Ambergris fragrance compound in US Patent Application Publication No. 2015/0260707 are Fold Increase. No response of value 10 or higher was shown.
Note that OR2L8 is a protein that is registered in GenBank as NM_001001963 and has an amino acid sequence (SEQ ID NO: 8) encoded by the first to 939th DNA of the base sequence represented by SEQ ID NO: 7. OR4K5 is registered in GenBank as NM_001005483 and is a protein consisting of an amino acid sequence (SEQ ID NO: 10) encoded by the first to 972nd DNAs of the base sequence shown in SEQ ID NO: 9.

Also, the receptor response of OR7A17 to various concentrations of compound (a) and the response of OR7C1 to various concentrations of compound (b) were measured by luciferase reporter gene assay. The results are shown in FIGS. 2A and B, respectively. OR7A17 showed a concentration-dependent response to compound (a) and OR7C1 to compound (b). On the other hand, no response was seen in the Mock test. That is, it was shown that OR7A17 specifically responds to compound (a) and OR7C1 to compound (b).

[Example 2]
Evaluation of response strength of OR7A17 and OR7C1 to Ambergris fragrance compound Compound (c) which is a fragrance compound exhibiting Ambergris notes using OR7A17 or OR7C1 which showed a strong response to Compound (a) or Compound (b) (Takasago Fragrance Co., Ltd.), Compound (d) (Takasago Fragrance Industrial Co., Ltd.), Compound (e) (obtained as a 50% Dowanol TPM diluted product from Givaudan), Compound (f) (10% TEC diluted product from Symrise) And the intensity of response to compound (g) (available as a 8.5% IPM dilution from Givaudan) was measured by luciferase reporter gene assay. The results are shown in FIGS. 3A, B, C, D and E, respectively. OR7A17 showed a concentration-dependent response to compound (c) and OR7C1 to compound (d), compound (e), compound (f) and compound (g). On the other hand, no response was seen in the Mock test. That is, it was shown that OR7A17 and OR7C1 respond specifically to various Ambergris fragrance compounds.

[Example 3]
Evaluation of response intensity of OR7A17 and OR7C1 to fragrance compounds with various fragrances The above-mentioned Ambergris note fragrance compound (a), compound (c), compound (d), compound (b), compound (e), compound ( In addition to f) and compound (g), the response intensities of OR7A17 and OR7C1 to the non-Amberlis note fragrance compounds listed in Table 1 of various fragrances for comparison were measured by a luciferase reporter gene assay. The concentration of each compound was set so as not to show cytotoxicity or non-receptor-specific response in the Mock test system.

 

 

In addition, the following were used for the non-Amberlis note fragrance compounds listed in Table 1.
D-Limone (Nacalai Tesque Corporation)
Cis-3-Hexenol (Nippon Zeon Corporation)
Melonal (Givaudan)
L-Menthol (Takasago International Corporation)
Linearyl acetate (Tokyo Chemical Industry Co., Ltd.)
Vanillin (Tokyo Chemical Industry Co., Ltd.)
Citronellol (Renessenz)
THESARON (registered trademark), (1R, 6S) -Ethyl 2,2,6-trimethylcyclohexanecarboxylate (Takasago International Corporation)
CALONE (registered trademark), 7-methylbenzo [b] [1,4] dioxepin-3-one (Firmenich)
HINDINOL (registered trademark), (R, E) -2-methyl-4- (2,2,3-trimethylcyclic-3-enyl) but-2-en-1-ol) (Takasago International Corporation)
JAVANOL (registered trademark), [(1R, 2S) -1-methyl-2-[[(1R, 3S, 5S) -1,2,2-trimethyl-3-bicyclo [3.1.0] hexenyl] methyl ] Cyclopropyl] methanol (Givaudan)
L-Muscone, (R) -3-methylcyclopentadecan-1-one (Takasago International Corporation)

The results are shown in FIG. OR7A17 and OR7C1 responded to any of the seven compounds exhibiting Ambergris notes and did not respond to the other aromatic compounds. That is, it was shown that OR7A17 and OR7C1 respond specifically to the Ambergris note fragrance compound.

By using the screening method of the present invention, candidate compounds exhibiting Ambergris notes can be selected from a large number of test compounds. The screening method of the present invention is expected to contribute to the development of a new fragrance material that exhibits Ambergris notes.

Claims (5)

A method for screening candidate compounds exhibiting Ambergris notes from test compounds using an olfactory receptor that exhibits responsiveness to a fragrance exhibiting Ambergris notes,
(I) A group comprising OR7A17, OR7C1, and a protein comprising an amino acid sequence having 80% or more identity with the amino acid sequence represented by SEQ ID NO: 2 or 4, and responsive to a fragrance exhibiting Ambergris notes Contacting a test compound with a more selected olfactory receptor and measuring a response of the olfactory receptor to the test compound;
(Ii) measuring the response of the olfactory receptor used in step (i) in the absence of the test compound;
(Iii) comparing the measurement results in steps (i) and (ii), and selecting a test compound with altered responsiveness as a candidate compound exhibiting Ambergris notes. The fragrance presenting Ambergris Note has the following formula:

The method according to claim 1, which is selected from the group consisting of compounds represented by: The response of the olfactory receptor is measured on a cell isolated from a living body expressing the olfactory receptor or on a cell in which the olfactory receptor is artificially expressed by genetic manipulation. Method. The method according to any one of claims 1 to 3, wherein the response of the olfactory receptor is measured by a reporter assay. When the Fold Increase value obtained by dividing the measurement result in step (i) by the measurement result in step (ii) is 2 or more, it is selected as a candidate compound that exhibits Ambergris note in step (iii). Item 5. The method according to any one of Items 1 to 4.

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