Screening method for odor suppression materials

Abstract

The present invention pertains to: a screening method for a material for suppressing at least one offensive odor selected from the group consisting of a 1-octen-3-one odor, a 1,5-octadien-3-one odor, a 1-octen-3-ol odor, and a 1,5-octadien-3-ol odor; and an offensive odor suppressing composition that reduces the offensiveness of age-related body odors and / or underarm odors; and a flavor composition for reducing off-flavors derived from food and beverages. By using the screening method according to the present invention, it is possible to select, from among a large number of test substances, respective candidate substances for materials for suppressing the 1-octen-3-one odor, the 1,5-octadien-3-one odor, the 1-octen-3-ol odor, and the 1,5-octadien-3-ol odor. The usage of an offensive odor suppressing composition that contains an active ingredient selected by using the screening method according to the present invention is expected to provide contribution in reducing the offensiveness of age-related body odors and / or underarm odors and reducing off-flavors derived from food and beverages.

Description

[Technical field]

[0001] The present invention relates to a method for screening an inhibitory material for one or more malodors selected from the group consisting of 1-octene-3-one odor, 1,5-octadien-3-one odor, 1-octene-3-ol odor and 1,5-octadien-3-ol odor, an malodor inhibitor composition that reduces the unpleasantness of aging odor and / or armpit odor, and a flavor composition that reduces off-flavors derived from foods and beverages. [Background technology]

[0002] There is a strong demand for more effective deodorization of unpleasant odors and bad smells in daily life to improve living environments, and in recent years, awareness of body odor has increased. Furthermore, it is known that food and beverages develop a deterioration odor (off-flavor) over time, which reduces the quality of the food and beverages.

[0003] Body odor is roughly divided into "smells from each part of the body" such as bad breath, foot odor, armpit odor, scalp odor, etc., and "smells that are a combination of odors emitted from the trunk of the body". It is known that "smells that are a combination of odors emitted from the trunk of the body" change with age, and the aging odor that is recognized in middle-aged and older people tends to be disliked very much. It has been reported that unsaturated aldehydes such as trans-2-nonenal and trans-2-octenal are the causative substances of this aging odor (Patent Document 1). It has also been reported that vinyl ketones such as 1-octen-3-one and 1,5-octadien-3-one have a very low odor threshold, a strong metallic odor, and are greatly involved in armpit odor (Non-Patent Document 1). Nonenal, 1-octen-3-one, 1,5-octadien-3-one, etc. are also known as off-flavors (Patent Documents 2 and 3, Non-Patent Document 2).

[0004] The possibility of selective deodorization technology based on the antagonistic mechanism of searching for olfactory receptors that respond to specific odorants and suppressing their activation with specific substances is becoming clear. For example, the olfactory receptor OR2W1 responds to hexanoic acid, which has a bad odor. Florhydral is a substance that inhibits the activity of this OR2W1, and it has been reported that florhydral suppresses the bad odor caused by hexanoic acid in sensory tests (Patent Document 4). [Prior art documents] [Patent documents]

[0005] [Patent Document 1] Japanese Patent Application Publication No. 11-286423 [Patent Document 2] JP 2017-153371 A [Patent Document 3] Patent No. 6168999 [Patent Document 4] JP 2012-50411 A [Non-patent literature]

[0006] [Non-Patent Document 1] "Analysis of the body odor generation mechanism and its countermeasures (1)" Satoru Iida et al., J. Soc. Cosmet. Chem. Jpn. Vol. 37, No. 3 (2003) [Non-Patent Document 2] "Odor Components of Edible Oils and Fats" Yasushi Endo, Journal of the Japan Oil Chemists' Society, Vol. 48, No. 10, (1999) Summary of the Invention [Problem to be solved by the invention]

[0007] It may be possible to search for olfactory receptors that respond to known malodor-causing substances and use them to screen candidate substances for malodor-suppressing materials. However, olfactory receptors that respond to one or more malodor-causing substances selected from the group consisting of 1-octen-3-one, 1,5-octadien-3-one, 1-octen-3-ol, and 1,5-octadien-3-ol have not been widely known so far. Under these circumstances, it is desired to provide a method for searching for olfactory receptors that respond to one or more malodor-causing substances selected from the group consisting of 1-octen-3-one, 1,5-octadien-3-one, 1-octen-3-ol, and 1,5-octadien-3-ol, and to provide a method for screening an inhibitory material for one or more malodors selected from the group consisting of 1-octen-3-one odor, 1,5-octadien-3-one odor, 1-octen-3-ol odor, and 1,5-octadien-3-ol odor, an malodor inhibitor composition that reduces the unpleasantness of aging odor and / or armpit odor, and a flavor composition that reduces off-flavors derived from foods and beverages. [Means for solving the problem]

[0008] The present inventors conducted intensive research to solve the above-mentioned problems and succeeded in identifying new olfactory receptors that respond to one or more malodor-causing substances selected from the group consisting of 1-octen-3-one, 1,5-octadien-3-one, 1-octen-3-ol, and 1,5-octadien-3-ol. The inventors further conducted research and discovered that by using these olfactory receptors, it is possible to evaluate and select materials that suppress aging odor and / or axillary odor and materials that reduce off-flavors by the masking effect of olfactory receptor antagonists, as well as materials that suppress one or more unpleasant odors selected from the group consisting of 1-octen-3-one odor, 1,5-octadien-3-one odor, 1-octen-3-ol odor, and 1,5-octadien-3-ol odor. They also discovered that a group of candidate antagonist compounds that suppress the responses of these olfactory receptors, including OR2C1 and OR4Q3, have excellent effects in suppressing aging odor and / or axillary odor, and in reducing off-flavors derived from foods and beverages.

[0009] That is, the present invention provides a method for screening inhibitory materials for one or more malodors selected from the group consisting of 1-octen-3-one odor, 1,5-octadien-3-one odor, 1-octen-3-ol odor and 1,5-octadien-3-ol odor, as shown below; an malodor inhibitor composition that reduces the unpleasantness of aging odor and / or armpit odor; and a flavor composition that reduces off-flavors derived from foods and beverages.

[0010] [1] A method for screening for an inhibitory material for one or more malodors selected from the group consisting of 1-octen-3-one odor, 1,5-octadien-3-one odor, 1-octen-3-ol odor, and 1,5-octadien-3-ol odor, comprising the steps of: (a) adding a test substance and one or more malodor-causing substances selected from the group consisting of 1-octen-3-one, 1,5-octadien-3-one, 1-octen-3-ol and 1,5-octadien-3-ol to at least one olfactory receptor polypeptide selected from the group consisting of OR2C1, OR4Q3, and (b) a polypeptide comprising an amino acid sequence having at least 80% identity with the amino acid sequence of any of the polypeptides of (a) and showing responsiveness to one or more malodor-causing substances selected from the group consisting of 1-octen-3-one, 1,5-octadien-3-one, 1-octen-3-ol and 1,5-octadien-3-ol; Measuring the response of the olfactory receptor polypeptide to one or more malodor-causing substances selected from the group consisting of 1-octen-3-one, 1,5-octadien-3-one, 1-octen-3-ol and 1,5-octadien-3-ol; and identifying a test substance that inhibits the response of the olfactory receptor polypeptide based on the measured response; The method includes: [2] The method according to [1], wherein the malodor is a 1-octen-3-one odor and / or a 1,5-octadien-3-one odor. [3] The method according to [1] or [2], in which the response of the olfactory receptor polypeptide is measured on cells isolated from an organism expressing the olfactory receptor polypeptide, or on cells in which the olfactory receptor polypeptide is artificially expressed by genetic manipulation. [4] The method according to any one of [1] to [3], wherein the response of the olfactory receptor polypeptide is measured by a reporter gene assay. Effect of the Invention

[0011] By using the method of the present invention, it is possible to screen candidate substances for suppressing one or more malodors selected from the group consisting of 1-octene-3-one odor, 1,5-octadien-3-one odor, 1-octene-3-ol odor, and 1,5-octadien-3-ol odor. It is expected that the screening method of the present invention can contribute to the development of suppressing materials for one or more malodors selected from the group consisting of 1-octene-3-one odor, 1,5-octadien-3-one odor, 1-octene-3-ol odor, and 1,5-octadien-3-ol odor. Furthermore, evaluating the odors of a large number of candidate substances using only the human sense of smell in order to develop new fragrance materials can lead to problems such as olfactory fatigue and individual differences, making it difficult to select appropriate candidate substances; however, the method of the present invention can eliminate or reduce such problems. Furthermore, according to a preferred embodiment of the present invention, by combining a group of candidate antagonist compounds that suppress the response of OR2C1 and / or OR4Q3 with a specific group of compounds that have traditionally been used as fragrances, etc., it is possible to provide a superior odor suppression effect than using only the group of candidate antagonist compounds. [Brief description of the drawings]

[0012] [Figure 1] FIG. 1 shows the results of measuring the response of the olfactory receptor OR2C1 to 1-octen-3-one. [Diagram 2] FIG. 1 shows the results of measuring the response of the olfactory receptor OR4Q3 to 1-octen-3-one. [Diagram 3] FIG. 1 shows the results of measuring the response of the olfactory receptor OR2C1 to 1,5-octadien-3-one. [Figure 4] FIG. 1 shows the results of measuring the response of the olfactory receptor OR4Q3 to 1,5-octadien-3-one. [Diagram 5] FIG. 1 shows the results of measuring the response of the olfactory receptor OR4Q3 to 1-octen-3-ol. [Figure 6] FIG. 1 shows the results of measuring the response of the olfactory receptor OR4Q3 to 1,5-octadien-3-ol. [Figure 7] FIG. 1 shows the results of the response-suppressing effect of the olfactory receptor OR2C1 to 1-octen-3-one by the addition of 2-methyl-4-(2,2,3-trimethylcyclopent-3-enyl)but-2-en-1-ol (Hindinol®). [Figure 8] FIG. 1 shows the results of the response-suppressing effect of the olfactory receptor OR2C1 to 1-octen-3-one by the addition of 1-(2,2,6-trimethylcyclohexyl)hexan-3-ol (Dextramber (registered trademark)). [Figure 9] FIG. 1 shows the results of the response-suppressing effect of the olfactory receptor OR2C1 to trans-2-nonenal by the addition of 2-methyl-4-(2,2,3-trimethylcyclopent-3-enyl)but-2-en-1-ol (Hindinol (registered trademark)). [Figure 10] FIG. 1 shows the results of the response-suppressing effect of the olfactory receptor OR2C1 to trans-2-nonenal by the addition of 1-(2,2,6-trimethylcyclohexyl)hexan-3-ol (Dextramber (registered trademark)). DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0013] The screening method, the malodor suppressing composition, and the flavor composition of the present invention will be specifically described below.

[0014] 1. Screening method for 1-octen-3-one odor suppressing materials The method for screening for a 1-octen-3-one odor suppressing material according to the present invention is a method for screening candidate substances for a 1-octen-3-one odor suppressing material from among test substances using an olfactory receptor polypeptide responsive to 1-octen-3-one, the method comprising the steps of: Adding a test substance and 1-octen-3-one to at least one olfactory receptor polypeptide selected from the group consisting of (a) OR2C1, OR4Q3, and (b) a polypeptide having an amino acid sequence having at least 80% identity to the amino acid sequence of any one of the polypeptides in (a) and exhibiting responsiveness to 1-octen-3-one; Measuring the response of the olfactory receptor polypeptide to the 1-octen-3-one; and Identifying a test substance that inhibits the response of the olfactory receptor polypeptide based on the measured response, i.e., identifying a test substance that inhibits the response of the olfactory receptor polypeptide based on the measured response as a candidate for a 1-octen-3-one odor suppressing material; The present invention is characterized in that it includes:

[0015] According to a preferred embodiment of the present invention, the screening method for a 1-octen-3-one odor suppressing material of the present invention comprises the steps of: (i) contacting an olfactory receptor polypeptide selected from the group consisting of (a) OR2C1, OR4Q3, and (b) a polypeptide having an amino acid sequence having 80% or more identity to the amino acid sequence of any one of the polypeptides of (a) and exhibiting responsiveness to 1-octen-3-one with 1-octen-3-one, and measuring the response of the olfactory receptor polypeptide to 1-octen-3-one; (ii) mixing a test substance with 1-octen-3-one and measuring the response of the olfactory receptor polypeptide used in step (i); (iii) comparing the measurement results in steps (i) and (ii) and selecting a test substance that reduces the response as a candidate substance for a 1-octen-3-one odor suppressing material; Includes.

[0016] The screening method for 1-octen-3-one odor suppressing materials of the present invention involves selecting candidate substances for 1-octen-3-one odor suppressing materials from among test substances, using as an index the responsiveness of the test substance to an olfactory receptor polypeptide selected from the group consisting of OR2C1, OR4Q3, and polypeptides that contain an amino acid sequence that has 80% or more identity to the amino acid sequences of these polypeptides and are responsive to odor-causing substances. In the screening method for 1-octen-3-one odor suppressing materials according to the present invention, the term "test substance" refers to a target for investigating the 1-octen-3-one odor suppressing effect, and is not particularly limited to a specific compound, composition, or mixture. The term "1-octen-3-one odor suppressing material" refers to a compound, composition, or mixture that can suppress the 1-octen-3-one odor, and is not particularly limited to a specific compound, composition, or mixture. The term "1-octen-3-one odor" refers to odors caused by 1-octen-3-one, and includes, for example, mushroom odor, iron rust odor, armpit odor, and off-flavor. Each step will be described below.

[0017] <Process (i)> In step (i), 1-octen-3-one is contacted with an olfactory receptor polypeptide selected from the group consisting of OR2C1, OR4Q3, and polypeptides that contain an amino acid sequence having 80% or more identity to the amino acid sequence of any of these polypeptides and are responsive to 1-octen-3-one, and the response of the olfactory receptor polypeptide to 1-octen-3-one is measured.

[0018] The olfactory receptor polypeptide used is an olfactory receptor polypeptide selected from the group consisting of OR2C1, OR4Q3, and polypeptides that contain an amino acid sequence that has 80% or more identity to the amino acid sequence of any of these polypeptides and are responsive to 1-octen-3-one. OR2C1 is registered in GenBank as NM_012368, and is a polypeptide consisting of an amino acid sequence (SEQ ID NO: 2) encoded by DNA from the 53rd to 991st positions of the nucleotide sequence shown in SEQ ID NO: 1. OR4Q3 is registered in GenBank as NM_172194, and is a polypeptide consisting of an amino acid sequence (sequence number 4) encoded by DNA having the base sequence shown in sequence number 3. Since these polypeptides selectively and strongly respond to 1-octen-3-one, it is expected that the screening method using OR2C1 and OR4Q3 will contribute to the development of materials that suppress the 1-octen-3-one odor.

[0019] As an olfactory receptor, an olfactory receptor polypeptide selected from the group consisting of polypeptides that contain an amino acid sequence that has 80% or more, preferably 85% or more, more preferably 90% or more, even more preferably 95% or more, and particularly preferably 98% or more identity to the amino acid sequence of these polypeptides (i.e., SEQ ID NO: 2 or 4) and are responsive to 1-octen-3-one may be used.

[0020] The olfactory receptor polypeptide may be used alone or in combination of two or more kinds.

[0021] In the present invention, the method of contacting an olfactory receptor polypeptide with 1-octen-3-one and measuring the response of the olfactory receptor polypeptide to 1-octen-3-one is not particularly limited. For example, the olfactory receptor polypeptide may be contacted with 1-octen-3-one on a cell isolated from a living organism expressing the olfactory receptor polypeptide and the response of the olfactory receptor polypeptide may be measured, or the olfactory receptor polypeptide may be contacted with 1-octen-3-one on a cell artificially expressed by genetic manipulation and the response of the olfactory receptor polypeptide may be measured. The time for contacting the olfactory receptor polypeptide with 1-octen-3-one depends on the concentration of 1-octen-3-one and the measurement method, so it cannot be generalized, but the response may be measured immediately after contact, usually 0 to 4 hours, preferably 2 to 4 hours. The same applies when a test substance is mixed with 1-octen-3-one and contacted with the olfactory receptor polypeptide.

[0022] Cells in which an olfactory receptor polypeptide is artificially expressed by genetic manipulation can be produced by transforming cells with a vector incorporating a gene encoding the olfactory receptor polypeptide.

[0023] 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 polypeptide. By incorporating the N-terminal 20 amino acid residues of bovine rhodopsin, the cell membrane expression of the olfactory receptor polypeptide can be promoted. Bovine rhodopsin is registered in GenBank as NM_001014890. Bovine rhodopsin is a polypeptide consisting of an amino acid sequence (SEQ ID NO: 6) encoded by DNA from the 1st to 1047th bases of the base sequence shown in SEQ ID NO: 5. In addition, instead of bovine rhodopsin, a polypeptide may be used that contains an amino acid sequence that has an identity of 80% or more, preferably 85% or more, more preferably 90% or more, even more preferably 95% or more, and particularly preferably 98% or more to the amino acid sequence shown in SEQ ID NO: 6, and that can promote cell membrane expression of an olfactory receptor polypeptide. In addition, amino acid residues of other polypeptides, not limited to bovine rhodopsin, may be used as long as they can promote cell membrane expression of the olfactory receptor polypeptide.

[0024] The method for measuring the response of an olfactory receptor polypeptide is not particularly limited, and any method used in the art can be used. For example, it is known that when an aroma compound binds to an olfactory receptor polypeptide, it activates intracellular G protein, which in turn activates adenylate cyclase to convert ATP to cyclic AMP (cAMP), thereby increasing the amount of cAMP in the cell. Therefore, the response of an olfactory receptor polypeptide can be measured by measuring the amount of cAMP. Methods for measuring the amount of cAMP include ELISA and reporter gene assays. Among these, it is preferable to measure the response of an olfactory receptor polypeptide by a reporter gene assay using a luminescent substance such as luciferase.

[0025] According to one embodiment of the present invention, the response of an olfactory receptor polypeptide may be evaluated using as an index the FoldIncrease value obtained by dividing the measurement result in the presence of 1-octen-3-one by the measurement result in the absence of 1-octen-3-one. For example, when measuring the response of an olfactory receptor polypeptide by a reporter gene assay method using a luminescent substance such as luciferase, the evaluation can be performed using 1-octen-3-one at a concentration that provides a FoldIncrease value of preferably 2 or more, more preferably 4 or more, and even more preferably 10 or more.

[0026] <Process (ii)> In step (ii), a test substance is mixed with 1-octen-3-one, and the response of the olfactory receptor used in step (i) is measured. The method for measuring the response of the olfactory receptor polypeptide can be the same as the method shown in step (i), except that 1-octen-3-one and a test substance are mixed and contacted with the olfactory receptor polypeptide. For example, the response of the olfactory receptor polypeptide may be measured on cells isolated from an organism expressing the olfactory receptor polypeptide, or the response of the olfactory receptor polypeptide may be measured on cells in which the olfactory receptor polypeptide is artificially expressed by genetic engineering. In order to appropriately compare the measurement results in steps (i) and (ii), it is preferable that the measurement conditions in steps (i) and (ii) are the same, except for the presence or absence of a test substance.

[0027] <Step (iii)> In step (iii), the measurement results in steps (i) and (ii) are compared, and the test substance showing a reduced response is selected as a candidate for the 1-octen-3-one odor suppressing material. In the present invention, when a reduction in response is observed by comparing the measurement results in steps (i) and (ii), the test substance used in step (ii) can be evaluated as a candidate substance for a 1-octen-3-one odor suppressing material.

[0028] As described above, candidate substances for 1-octen-3-one odor suppressing materials can be screened from among the test substances. According to the present invention, candidate substances for 1-octen-3-one odor suppressing materials can be selected from a large number of test substances without problems such as olfactory fatigue and individual differences caused by sensory evaluation performed by human olfaction. The selected substances can be used as candidate substances for 1-octen-3-one odor suppressing materials. Based on the selected substances, modifications, etc. can be carried out as necessary to develop novel compounds with optimal odors. Furthermore, the selected substances can be blended with other fragrance materials to suppress the 1-octen-3-one odor and develop fragrance materials with optimal odors. Use of the screening method of the present invention can contribute to the development of new fragrance materials for 1-octen-3-one odor suppressing materials.

[0029] 2. Screening method for 1,5-octadien-3-one odor suppressing materials The method for screening for a 1,5-octadien-3-one odor suppressing material according to the present invention is a method for screening candidate substances for a 1,5-octadien-3-one odor suppressing material from among test substances using an olfactory receptor polypeptide responsive to 1,5-octadien-3-one, the method comprising the steps of: Adding a test substance and 1,5-octadien-3-one to at least one olfactory receptor polypeptide selected from the group consisting of (a) OR2C1, OR4Q3, and (b) a polypeptide having an amino acid sequence having at least 80% identity to the amino acid sequence of any of the polypeptides in (a) and exhibiting responsiveness to 1,5-octadien-3-one; Measuring the response of the olfactory receptor polypeptide to the 1,5-octadien-3-one; and Identifying a test substance that inhibits the response of the olfactory receptor polypeptide based on the measured response, i.e., identifying a test substance that inhibits the response of the olfactory receptor polypeptide based on the measured response as a candidate 1,5-octadien-3-one odor suppressing material; The present invention is characterized in that it includes:

[0030] According to a preferred embodiment of the present invention, the screening method for a 1,5-octadien-3-one odor suppressing material of the present invention comprises the steps of: (i) contacting an olfactory receptor polypeptide selected from the group consisting of (a) OR2C1, OR4Q3, and (b) a polypeptide having an amino acid sequence having 80% or more identity to the amino acid sequence of any of the polypeptides of (a) and exhibiting responsiveness to 1,5-octadien-3-one with 1,5-octadien-3-one, and measuring the response of the olfactory receptor polypeptide to 1,5-octadien-3-one; (ii) mixing a test substance with 1,5-octadien-3-one and measuring the response of the olfactory receptor polypeptide used in step (i); (iii) comparing the measurement results in steps (i) and (ii) and selecting a test substance that reduces the response as a candidate substance for a 1,5-octadien-3-one odor suppressing material; Includes.

[0031] The screening method for 1,5-octadien-3-one odor suppressing materials of the present invention involves selecting candidate substances for 1,5-octadien-3-one odor suppressing materials from among test substances, using as an index the responsiveness of the test substance to an olfactory receptor polypeptide selected from the group consisting of OR2C1, OR4Q3, and polypeptides that contain an amino acid sequence that has 80% or more identity with the amino acid sequences of these polypeptides and are responsive to odor-causing substances. In the screening method for 1,5-octadien-3-one odor suppressing materials according to the present invention, the term "test substance" refers to a target for investigating the effect of suppressing 1,5-octadien-3-one odor, and is not particularly limited to a specific compound, composition, or mixture. The term "1,5-octadien-3-one odor suppressing material" refers to a compound, composition, or mixture that can suppress 1,5-octadien-3-one odor, and is not particularly limited to a specific compound, composition, or mixture. The term "1,5-octadien-3-one odor" refers to odors caused by 1,5-octadien-3-one, and includes, for example, mushroom odor, iron rust odor, armpit odor, and off-flavor. Each step will be described below.

[0032] <Process (i)> In step (i), an olfactory receptor polypeptide selected from the group consisting of OR2C1, OR4Q3, and polypeptides that contain an amino acid sequence that has 80% or more identity to the amino acid sequence of any of these polypeptides and are responsive to 1,5-octadien-3-one is contacted with 1,5-octadien-3-one, and the response of the olfactory receptor polypeptide to 1,5-octadien-3-one is measured.

[0033] The olfactory receptor polypeptide used is an olfactory receptor polypeptide selected from the group consisting of OR2C1, OR4Q3, and polypeptides that contain an amino acid sequence that has 80% or more identity to the amino acid sequence of any of these polypeptides and are responsive to 1,5-octadien-3-one. OR2C1 is registered in GenBank as NM_012368, and is a polypeptide consisting of an amino acid sequence (SEQ ID NO: 2) encoded by DNA from the 53rd to 991st positions of the nucleotide sequence shown in SEQ ID NO: 1. OR4Q3 is registered in GenBank as NM_172194, and is a polypeptide consisting of an amino acid sequence (sequence number 4) encoded by DNA having the base sequence shown in sequence number 3. Since these polypeptides selectively and strongly respond to 1,5-octadien-3-one, it is expected that the screening method using OR2C1 and OR4Q3 will contribute to the development of materials that suppress the 1,5-octadien-3-one odor.

[0034] As an olfactory receptor, an olfactory receptor polypeptide selected from the group consisting of polypeptides that contain an amino acid sequence that has 80% or more, preferably 85% or more, more preferably 90% or more, even more preferably 95% or more, and particularly preferably 98% or more identity to the amino acid sequence of these polypeptides (i.e., SEQ ID NO: 2 or 4) and that are responsive to 1,5-octadien-3-one may be used.

[0035] The olfactory receptor polypeptide may be used alone or in combination of two or more kinds.

[0036] In the present invention, the method of contacting an olfactory receptor polypeptide with 1,5-octadien-3-one and measuring the response of the olfactory receptor polypeptide to 1,5-octadien-3-one is not particularly limited. For example, the olfactory receptor polypeptide may be contacted with 1,5-octadien-3-one on a cell isolated from a living organism expressing the olfactory receptor polypeptide and the response of the olfactory receptor polypeptide may be measured, or the olfactory receptor polypeptide may be contacted with 1,5-octadien-3-one on a cell in which the olfactory receptor polypeptide is artificially expressed by genetic engineering and the response of the olfactory receptor polypeptide may be measured. The time for which the olfactory receptor polypeptide is contacted with 1,5-octadien-3-one depends on the concentration of 1,5-octadien-3-one and the measurement method, so it cannot be generalized, but the response may be measured immediately after contact, usually for 0 to 4 hours, preferably for 2 to 4 hours. The same applies when a test substance is mixed with 1,5-octadien-3-one and contacted with the olfactory receptor polypeptide.

[0037] Cells in which an olfactory receptor polypeptide is artificially expressed by genetic manipulation can be produced by transforming cells with a vector incorporating a gene encoding the olfactory receptor polypeptide.

[0038] 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 polypeptide. By incorporating the N-terminal 20 amino acid residues of bovine rhodopsin, the cell membrane expression of the olfactory receptor polypeptide can be promoted. Bovine rhodopsin is registered in GenBank as NM_001014890. Bovine rhodopsin is a polypeptide consisting of an amino acid sequence (SEQ ID NO: 6) encoded by DNA from the 1st to 1047th bases of the base sequence shown in SEQ ID NO: 5. In addition, instead of bovine rhodopsin, a polypeptide may be used that contains an amino acid sequence that has an identity of 80% or more, preferably 85% or more, more preferably 90% or more, even more preferably 95% or more, and particularly preferably 98% or more to the amino acid sequence shown in SEQ ID NO: 6, and that can promote cell membrane expression of an olfactory receptor polypeptide. In addition, amino acid residues of other polypeptides, not limited to bovine rhodopsin, may be used as long as they can promote cell membrane expression of the olfactory receptor polypeptide.

[0039] The method for measuring the response of an olfactory receptor polypeptide is not particularly limited, and any method used in the art can be used. For example, it is known that when an aroma compound binds to an olfactory receptor polypeptide, it activates intracellular G protein, which in turn activates adenylate cyclase to convert ATP to cyclic AMP (cAMP), thereby increasing the amount of cAMP in the cell. Therefore, the response of an olfactory receptor polypeptide can be measured by measuring the amount of cAMP. Methods for measuring the amount of cAMP include ELISA and reporter gene assays. Among these, it is preferable to measure the response of an olfactory receptor polypeptide by a reporter gene assay using a luminescent substance such as luciferase.

[0040] According to one embodiment of the present invention, the response of an olfactory receptor polypeptide may be evaluated using as an index the FoldIncrease value obtained by dividing the measurement result in the presence of 1,5-octadien-3-one by the measurement result in the absence of 1,5-octadien-3-one. For example, when measuring the response of an olfactory receptor polypeptide by a reporter gene assay method using a luminescent substance such as luciferase, the evaluation can be performed using a concentration of 1,5-octadien-3-one that provides a FoldIncrease value of preferably 2 or more, more preferably 4 or more, and even more preferably 10 or more.

[0041] <Process (ii)> In step (ii), a test substance is mixed with 1,5-octadien-3-one, and the response of the olfactory receptor used in step (i) is measured. The method for measuring the response of the olfactory receptor polypeptide can be the same as the method shown in step (i), except that 1,5-octadien-3-one and the test substance are mixed and contacted with the olfactory receptor polypeptide. For example, the response of the olfactory receptor polypeptide may be measured on cells isolated from an organism expressing the olfactory receptor polypeptide, or the response of the olfactory receptor polypeptide may be measured on cells in which the olfactory receptor polypeptide is artificially expressed by genetic engineering. In order to appropriately compare the measurement results in steps (i) and (ii), it is preferable that the measurement conditions in steps (i) and (ii) are the same, except for the presence or absence of the test substance.

[0042] <Step (iii)> In step (iii), the measurement results in steps (i) and (ii) are compared, and the test substance which gives a reduced response is selected as a candidate for the 1,5-octadien-3-one odor suppressing material. In the present invention, when a reduction in response is observed by comparing the measurement results in steps (i) and (ii), the test substance used in step (ii) can be evaluated as a candidate substance for a 1,5-octadien-3-one odor suppressing material.

[0043] As described above, candidate substances for 1,5-octadien-3-one odor suppressing materials can be screened from among the test substances. According to the present invention, candidate substances for 1,5-octadien-3-one odor suppressing materials can be selected from a large number of test substances without problems such as olfactory fatigue and individual differences caused by sensory evaluation using human olfaction. The selected substances can be used as candidate substances for 1,5-octadien-3-one odor suppressing materials. Based on the selected substances, modifications, etc. can be carried out as necessary to develop novel compounds with optimal odors. Furthermore, the selected substances can be blended with other fragrance materials to suppress the 1,5-octadien-3-one odor and develop fragrance materials with optimal odors. Use of the screening method of the present invention can contribute to the development of new fragrance materials for 1,5-octadien-3-one odor suppressing materials.

[0044] 3. Screening method for 1-octen-3-ol odor suppressing materials The method for screening for a 1-octen-3-ol odor-suppressing material according to the present invention is a method for screening candidate substances for a 1-octen-3-ol odor-suppressing material from among test substances using an olfactory receptor polypeptide responsive to 1-octen-3-ol, the method comprising the steps of: Adding a test substance and 1-octen-3-ol to at least one olfactory receptor polypeptide selected from the group consisting of (a) OR2C1, OR4Q3, and (b) a polypeptide comprising an amino acid sequence having at least 80% identity to the amino acid sequence of any of the polypeptides in (a) and exhibiting responsiveness to 1-octen-3-ol; Measuring the response of the olfactory receptor polypeptide to the 1-octen-3-ol; and Identifying a test substance that inhibits the response of the olfactory receptor polypeptide based on the measured response, i.e., identifying a test substance that inhibits the response of the olfactory receptor polypeptide based on the measured response as a candidate substance for a 1-octen-3-ol odor-inhibiting material; The present invention is characterized in that it includes:

[0045] According to a preferred embodiment of the present invention, the screening method for a 1-octen-3-ol odor suppressing material of the present invention comprises the steps of: (i) contacting 1-octen-3-ol with an olfactory receptor polypeptide selected from the group consisting of (a) OR2C1, OR4Q3, and (b) a polypeptide that has an amino acid sequence having 80% or more identity to the amino acid sequence of any of the polypeptides in (a) and is responsive to 1-octen-3-ol, and measuring the response of the olfactory receptor polypeptide to 1-octen-3-ol; (ii) mixing a test substance with 1-octen-3-ol and measuring the response of the olfactory receptor polypeptide used in step (i); (iii) comparing the measurement results in steps (i) and (ii) and selecting a test substance that reduces the response as a candidate substance for a 1-octen-3-ol odor suppressing material; Includes.

[0046] The screening method for 1-octen-3-ol odor-suppressing materials of the present invention involves selecting candidate substances for 1-octen-3-ol odor-suppressing materials from among test substances, using as an index the responsiveness of the test substance to an olfactory receptor polypeptide selected from the group consisting of OR2C1, OR4Q3, and polypeptides that contain an amino acid sequence that has 80% or more identity with the amino acid sequences of these polypeptides and are responsive to odor-causing substances. In the screening method for 1-octen-3-ol odor suppressing materials according to the present invention, the term "test substance" refers to a target for investigating the 1-octen-3-ol odor suppressing effect, and is not particularly limited, and refers to a compound, composition, or mixture. In addition, the term "1-octen-3-ol odor suppressing material" refers to a compound, composition, or mixture that can suppress the 1-octen-3-ol odor, and is not particularly limited. In addition, the term "1-octen-3-ol odor" refers to an odor caused by 1-octen-3-ol, and includes, for example, mushroom odor, iron rust odor, armpit odor, and off-flavor. Each step will be described below.

[0047] <Process (i)> In step (i), 1-octen-3-ol is contacted with an olfactory receptor polypeptide selected from the group consisting of OR2C1, OR4Q3, and polypeptides that contain an amino acid sequence having 80% or more identity to the amino acid sequence of any of these polypeptides and are responsive to 1-octen-3-ol, and the response of the olfactory receptor polypeptide to 1-octen-3-ol is measured.

[0048] The olfactory receptor polypeptide used is an olfactory receptor polypeptide selected from the group consisting of OR2C1, OR4Q3, and polypeptides that contain an amino acid sequence that has 80% or more identity to the amino acid sequence of any of these polypeptides and are responsive to 1-octen-3-ol. OR2C1 is registered in GenBank as NM_012368, and is a polypeptide consisting of an amino acid sequence (SEQ ID NO: 2) encoded by DNA from the 53rd to 991st positions of the nucleotide sequence shown in SEQ ID NO: 1. OR4Q3 is registered in GenBank as NM_172194, and is a polypeptide consisting of an amino acid sequence (sequence number 4) encoded by DNA having the base sequence shown in sequence number 3. Since these polypeptides selectively and strongly respond to 1-octen-3-ol, it is expected that the screening method using OR2C1 and OR4Q3 will contribute to the development of materials that suppress the 1-octen-3-ol odor.

[0049] As an olfactory receptor, an olfactory receptor polypeptide selected from the group consisting of polypeptides that contain an amino acid sequence that has 80% or more, preferably 85% or more, more preferably 90% or more, even more preferably 95% or more, and particularly preferably 98% or more identity to the amino acid sequence of these polypeptides (i.e., SEQ ID NO: 2 or 4) and are responsive to 1-octen-3-ol may be used.

[0050] The olfactory receptor polypeptide may be used alone or in combination of two or more kinds.

[0051] In the present invention, the method of contacting an olfactory receptor polypeptide with 1-octen-3-ol and measuring the response of the olfactory receptor polypeptide to 1-octen-3-ol is not particularly limited. For example, the olfactory receptor polypeptide may be contacted with 1-octen-3-ol on a cell isolated from a living organism expressing the olfactory receptor polypeptide and the response of the olfactory receptor polypeptide may be measured, or the olfactory receptor polypeptide may be contacted with 1-octen-3-ol on a cell in which the olfactory receptor polypeptide is artificially expressed by genetic manipulation and the response of the olfactory receptor polypeptide may be measured. The time for which the olfactory receptor polypeptide is contacted with 1-octen-3-ol depends on the concentration of 1-octen-3-ol and the measurement method, so it cannot be generally stated, but the response may be measured immediately after contact, usually for 0 to 4 hours, preferably for 2 to 4 hours. The same applies when a test substance is mixed with 1-octen-3-ol and contacted with the olfactory receptor polypeptide.

[0052] Cells in which an olfactory receptor polypeptide is artificially expressed by genetic manipulation can be produced by transforming cells with a vector incorporating a gene encoding the olfactory receptor polypeptide.

[0053] 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 polypeptide. By incorporating the N-terminal 20 amino acid residues of bovine rhodopsin, the cell membrane expression of the olfactory receptor polypeptide can be promoted. Bovine rhodopsin is registered in GenBank as NM_001014890. Bovine rhodopsin is a polypeptide consisting of an amino acid sequence (SEQ ID NO: 6) encoded by DNA from the 1st to 1047th bases of the base sequence shown in SEQ ID NO: 5. In addition, instead of bovine rhodopsin, a polypeptide may be used that contains an amino acid sequence that has an identity of 80% or more, preferably 85% or more, more preferably 90% or more, even more preferably 95% or more, and particularly preferably 98% or more to the amino acid sequence shown in SEQ ID NO: 6, and that can promote cell membrane expression of an olfactory receptor polypeptide. In addition, amino acid residues of other polypeptides, not limited to bovine rhodopsin, may be used as long as they can promote cell membrane expression of the olfactory receptor polypeptide.

[0054] The method for measuring the response of an olfactory receptor polypeptide is not particularly limited, and any method used in the art can be used. For example, it is known that when an aroma compound binds to an olfactory receptor polypeptide, it activates intracellular G protein, which in turn activates adenylate cyclase to convert ATP to cyclic AMP (cAMP), thereby increasing the amount of cAMP in the cell. Therefore, the response of an olfactory receptor polypeptide can be measured by measuring the amount of cAMP. Methods for measuring the amount of cAMP include ELISA and reporter gene assays. Among these, it is preferable to measure the response of an olfactory receptor polypeptide by a reporter gene assay using a luminescent substance such as luciferase.

[0055] According to one embodiment of the present invention, the response of an olfactory receptor polypeptide may be evaluated using as an index the FoldIncrease value obtained by dividing the measurement result in the presence of 1-octen-3-ol by the measurement result in the absence of 1-octen-3-ol. For example, when the response of an olfactory receptor polypeptide is measured by a reporter gene assay method using a luminescent substance such as luciferase, the evaluation can be performed using 1-octen-3-ol at a concentration that provides a FoldIncrease value of preferably 2 or more, more preferably 4 or more, and even more preferably 10 or more.

[0056] <Process (ii)> In step (ii), a test substance is mixed with 1-octen-3-ol, and the response of the olfactory receptor used in step (i) is measured. The method for measuring the response of the olfactory receptor polypeptide can be the same as the method shown in step (i), except that 1-octen-3-ol and a test substance are mixed and contacted with the olfactory receptor polypeptide. For example, the response of the olfactory receptor polypeptide may be measured on cells isolated from an organism expressing the olfactory receptor polypeptide, or the response of the olfactory receptor polypeptide may be measured on cells in which the olfactory receptor polypeptide is artificially expressed by genetic engineering. In order to appropriately compare the measurement results in steps (i) and (ii), it is preferable that the measurement conditions in steps (i) and (ii) are the same, except for the presence or absence of a test substance.

[0057] <Step (iii)> In step (iii), the measurement results in steps (i) and (ii) are compared, and the test substance showing a reduced response is selected as a candidate substance for the 1-octen-3-ol odor suppressing material. In the present invention, when a reduction in response is observed by comparing the measurement results in steps (i) and (ii), the test substance used in step (ii) can be evaluated as a candidate substance for a 1-octen-3-ol odor suppressing material.

[0058] As described above, candidate substances for 1-octen-3-ol odor suppressing materials can be screened from among the test substances. According to the present invention, candidate substances for 1-octen-3-ol odor suppressing materials can be selected from a large number of test substances without problems such as olfactory fatigue and individual differences caused by sensory evaluation performed by human olfaction. The selected substances can be used as candidate substances for 1-octen-3-ol odor suppressing materials. Based on the selected substances, modifications, etc. can be carried out as necessary to develop novel compounds with optimal odors. Furthermore, the selected substances can be blended with other fragrance materials to suppress the 1-octen-3-ol odor and develop fragrance materials with optimal odors. Use of the screening method of the present invention can contribute to the development of new fragrance materials for 1-octen-3-ol odor suppressing materials.

[0059] 4. Screening method for 1,5-octadien-3-ol odor suppressing materials The method for screening for a 1,5-octadien-3-ol odor-suppressing material according to the present invention is a method for screening candidate substances for a 1,5-octadien-3-ol odor-suppressing material from among test substances using an olfactory receptor polypeptide responsive to 1,5-octadien-3-ol, the method comprising the steps of: Adding a test substance and 1,5-octadien-3-ol to at least one olfactory receptor polypeptide selected from the group consisting of (a) OR2C1, OR4Q3, and (b) a polypeptide having an amino acid sequence having at least 80% identity to the amino acid sequence of any of the polypeptides in (a) and exhibiting responsiveness to 1,5-octadien-3-ol; Measuring the response of the olfactory receptor polypeptide to the 1,5-octadien-3-ol; and Identifying a test substance that inhibits the response of the olfactory receptor polypeptide based on the measured response, i.e., identifying a test substance that inhibits the response of the olfactory receptor polypeptide based on the measured response as a candidate 1,5-octadien-3-ol odor-inhibiting material; The present invention is characterized in that it includes:

[0060] According to a preferred embodiment of the present invention, the screening method for a 1,5-octadien-3-ol odor suppressing material of the present invention comprises the steps of: (i) contacting an olfactory receptor polypeptide selected from the group consisting of (a) OR2C1, OR4Q3, and (b) a polypeptide that contains an amino acid sequence having 80% or more identity to the amino acid sequence of any of the polypeptides in (a) and is responsive to 1,5-octadien-3-ol with 1,5-octadien-3-ol, and measuring the response of the olfactory receptor polypeptide to 1,5-octadien-3-ol; (ii) mixing a test substance with 1,5-octadien-3-ol and measuring the response of the olfactory receptor polypeptide used in step (i); (iii) comparing the measurement results in steps (i) and (ii) and selecting a test substance that reduces the response as a candidate substance for a 1,5-octadien-3-ol odor suppressing material; Includes.

[0061] The screening method for 1,5-octadien-3-ol odor-suppressing materials of the present invention involves selecting candidate substances for 1,5-octadien-3-ol odor-suppressing materials from among test substances, using as an index the responsiveness of the test substance to an olfactory receptor polypeptide selected from the group consisting of OR2C1, OR4Q3, and polypeptides that contain an amino acid sequence that has 80% or more identity with the amino acid sequences of these polypeptides and are responsive to odor-causing substances. In the screening method for 1,5-octadien-3-ol odor suppressing materials according to the present invention, the term "test substance" refers to a target for investigating the effect of suppressing 1,5-octadien-3-ol odor, and refers to a compound, composition, or mixture, without any particular limitation. The term "1,5-octadien-3-ol odor suppressing material" refers to a compound, composition, or mixture that can suppress 1,5-octadien-3-ol odor, without any particular limitation. The term "1,5-octadien-3-ol odor" refers to odors caused by 1,5-octadien-3-ol, and includes, for example, mushroom odor, iron rust odor, armpit odor, and off-flavor. Each step will be described below.

[0062] <Process (i)> In step (i), an olfactory receptor polypeptide selected from the group consisting of OR2C1, OR4Q3, and polypeptides that contain an amino acid sequence that has 80% or more identity to the amino acid sequence of any of these polypeptides and are responsive to 1,5-octadien-3-ol is contacted with 1,5-octadien-3-ol, and the response of the olfactory receptor polypeptide to 1,5-octadien-3-ol is measured.

[0063] The olfactory receptor polypeptide used is an olfactory receptor polypeptide selected from the group consisting of OR2C1, OR4Q3, and polypeptides that contain an amino acid sequence that has 80% or more identity to the amino acid sequence of any of these polypeptides and are responsive to 1,5-octadien-3-ol. OR2C1 is registered in GenBank as NM_012368, and is a polypeptide consisting of an amino acid sequence (SEQ ID NO: 2) encoded by DNA from the 53rd to 991st positions of the nucleotide sequence shown in SEQ ID NO: 1. OR4Q3 is registered in GenBank as NM_172194, and is a polypeptide consisting of an amino acid sequence (sequence number 4) encoded by DNA having the base sequence shown in sequence number 3. Since these polypeptides selectively and strongly respond to 1,5-octadien-3-ol, it is expected that the screening method using OR2C1 and OR4Q3 will contribute to the development of materials that suppress the odor of 1,5-octadien-3-ol.

[0064] As an olfactory receptor, an olfactory receptor polypeptide selected from the group consisting of polypeptides that contain an amino acid sequence that has 80% or more, preferably 85% or more, more preferably 90% or more, even more preferably 95% or more, and particularly preferably 98% or more identity to the amino acid sequence of these polypeptides (i.e., SEQ ID NO: 2 or 4) and are responsive to 1,5-octadien-3-ol may be used.

[0065] The olfactory receptor polypeptide may be used alone or in combination of two or more kinds.

[0066] In the present invention, the method of contacting an olfactory receptor polypeptide with 1,5-octadien-3-ol and measuring the response of the olfactory receptor polypeptide to 1,5-octadien-3-ol is not particularly limited. For example, the olfactory receptor polypeptide may be contacted with 1,5-octadien-3-ol on a cell isolated from a living organism expressing the olfactory receptor polypeptide and the response of the olfactory receptor polypeptide may be measured, or the olfactory receptor polypeptide may be contacted with 1,5-octadien-3-ol on a cell in which the olfactory receptor polypeptide is artificially expressed by genetic manipulation and the response of the olfactory receptor polypeptide may be measured. The time for which the olfactory receptor polypeptide is contacted with 1,5-octadien-3-ol depends on the concentration of 1,5-octadien-3-ol and the measurement method, so it cannot be generalized, but the response may be measured immediately after contact, usually for 0 to 4 hours, preferably for 2 to 4 hours. The same applies when a test substance is mixed with 1,5-octadien-3-ol and contacted with the olfactory receptor polypeptide.

[0067] Cells in which an olfactory receptor polypeptide is artificially expressed by genetic manipulation can be produced by transforming cells with a vector incorporating a gene encoding the olfactory receptor polypeptide.

[0068] 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 polypeptide. By incorporating the N-terminal 20 amino acid residues of bovine rhodopsin, the cell membrane expression of the olfactory receptor polypeptide can be promoted. Bovine rhodopsin is registered in GenBank as NM_001014890. Bovine rhodopsin is a polypeptide consisting of an amino acid sequence (SEQ ID NO: 6) encoded by DNA from the 1st to 1047th bases of the base sequence shown in SEQ ID NO: 5. In addition, instead of bovine rhodopsin, a polypeptide may be used that contains an amino acid sequence that has an identity of 80% or more, preferably 85% or more, more preferably 90% or more, even more preferably 95% or more, and particularly preferably 98% or more to the amino acid sequence shown in SEQ ID NO: 6, and that can promote cell membrane expression of an olfactory receptor polypeptide. In addition, amino acid residues of other polypeptides, not limited to bovine rhodopsin, may be used as long as they can promote cell membrane expression of the olfactory receptor polypeptide.

[0069] The method for measuring the response of an olfactory receptor polypeptide is not particularly limited, and any method used in the art can be used. For example, it is known that when an aroma compound binds to an olfactory receptor polypeptide, it activates intracellular G protein, which in turn activates adenylate cyclase to convert ATP to cyclic AMP (cAMP), thereby increasing the amount of cAMP in the cell. Therefore, the response of an olfactory receptor polypeptide can be measured by measuring the amount of cAMP. Methods for measuring the amount of cAMP include ELISA and reporter gene assays. Among these, it is preferable to measure the response of an olfactory receptor polypeptide by a reporter gene assay using a luminescent substance such as luciferase.

[0070] According to one embodiment of the present invention, the response of an olfactory receptor polypeptide may be evaluated using as an index the FoldIncrease value obtained by dividing the measurement result in the presence of 1,5-octadien-3-ol by the measurement result in the absence of 1,5-octadien-3-ol. For example, when measuring the response of an olfactory receptor polypeptide by a reporter gene assay method using a luminescent substance such as luciferase, the evaluation can be performed using 1,5-octadien-3-ol at a concentration that provides a FoldIncrease value of preferably 2 or more, more preferably 4 or more, and even more preferably 10 or more.

[0071] <Process (ii)> In step (ii), a test substance is mixed with 1,5-octadien-3-ol, and the response of the olfactory receptor used in step (i) is measured. The method for measuring the response of the olfactory receptor polypeptide can be the same as the method shown in step (i), except that 1,5-octadien-3-ol and a test substance are mixed and contacted with the olfactory receptor polypeptide. For example, the response of the olfactory receptor polypeptide may be measured on cells isolated from an organism expressing the olfactory receptor polypeptide, or the response of the olfactory receptor polypeptide may be measured on cells in which the olfactory receptor polypeptide is artificially expressed by genetic engineering. In order to appropriately compare the measurement results in steps (i) and (ii), it is preferable that the measurement conditions in steps (i) and (ii) are the same, except for the presence or absence of a test substance.

[0072] <Step (iii)> In step (iii), the measurement results in steps (i) and (ii) are compared, and the test substance showing a reduced response is selected as a candidate substance for the 1,5-octadien-3-ol odor suppressing material. In the present invention, when a reduction in response is observed by comparing the measurement results in steps (i) and (ii), the test substance used in step (ii) can be evaluated as a candidate substance for a 1,5-octadien-3-ol odor suppressing material.

[0073] As described above, candidate substances for 1,5-octadien-3-ol odor suppressing materials can be screened from among the test substances. According to the present invention, candidate substances for 1,5-octadien-3-ol odor suppressing materials can be selected from a large number of test substances without problems such as olfactory fatigue and individual differences caused by sensory evaluation performed by human olfaction. The selected substances can be used as candidate substances for 1,5-octadien-3-ol odor suppressing materials. Based on the selected substances, modifications, etc. can be carried out as necessary to develop novel compounds with optimal odors. Furthermore, the selected substances can be blended with other fragrance materials to suppress the 1,5-octadien-3-ol odor and develop fragrance materials with optimal odors. Use of the screening method of the present invention can contribute to the development of new fragrance materials for 1,5-octadien-3-ol odor suppressing materials.

[0074] 5. Odor Control Composition By using the screening method of the present invention, an appropriate malodor suppression material can be selected depending on the malodor-causing substance, and by using the malodor suppression material as an active ingredient, a malodor suppression composition suitable for the purpose can be obtained. In one embodiment of the present invention, the malodor suppressing composition contains, as an active ingredient, at least one compound selected from the following group (A): Examples of group (A) include 2-methyl-4-(2,2,3-trimethylcyclopent-3-enyl)but-2-en-1-ol (Hindinol (registered trademark)), 1-(2,2,6-trimethylcyclohexyl)hexan-3-ol (Dextramber (registered trademark)), 1,4-dioxacycloheptadecane-5,17-dione, eucalyptus oil, patchouli oil, vetiver oil, etc. The amount of group (A) in the malodor suppressing composition of the present invention is not strictly limited, and is preferably 0.0001% by weight or more, more preferably 0.001 to 20% by weight, and even more preferably 0.01 to 10% by weight.

[0075] The malodor suppressing composition containing the above (A) group can particularly suppress aged odor and / or armpit odor, which are malodors composed of at least one selected from the group consisting of 1-octene-3-one odor and 1,5-octadiene-3-one odor. Also, the malodor suppressing composition containing the above (A) group can suppress aged odor and / or armpit odor, which are malodors composed of at least one selected from the group consisting of trans-2-nonenal odor and trans-2-octenal odor.

[0076] The malodor suppression composition of the present invention may contain known fragrances or conventional additives as described below within a range that does not impair the effects of the present invention, i.e., within a quantitative and qualitative range that can eliminate the unpleasant sensations associated with aging odor and / or underarm odor. Examples of known fragrances include, for example, hydrocarbons such as α-pinene, limonene, and cedrene; aliphatic saturated or unsaturated alcohols such as 1-octen-3-ol and nonanol; saturated or unsaturated chain terpene alcohols such as myrcenol, tetrahydrolinalool, and nerol; cyclic terpene alcohols such as α-terpineol and bornylmethoxycyclohexanol; sesquiterpene alcohols such as farnesol; aromatic alcohols such as γ-phenylpropyl alcohol and dimethylbenzylcarbinol; aliphatic aldehydes, terpene aldehydes, aromatic aldehydes, and aliphatic ketones such as Iso-E-Super (7-acetyl-1,2,3,4,5,6,7,8-octahydro-1,1,6,7-tetramethylnaphthalene); terpene cyclic ketones, cyclic ketones, aliphatic esters, acetates, propionates, butyrates, benzoates, phenyl acetates, salicylates, and anthranilates. The amount of these known fragrances to be blended in the present malodor suppressing composition is not particularly limited and can be appropriately selected depending on the type of fragrance to be blended, the purpose, etc.

[0077] The malodor suppressing composition of the present invention can be incorporated into various products, and cosmetics are one of the preferred products. The above-mentioned malodor suppressing composition of the present invention can be incorporated into cosmetics and the like together with conventional additives depending on the form and formulation.

[0078] Examples of conventional additives include known ultraviolet absorbers, cooling agents such as menthol and methyl salicylate, and skin irritants such as capsicum tincture and nonyl vanillylamide, etc. The amount of these conventional additives to be added is not particularly limited, but is generally in the range of 0.005 to 5.0% by weight based on the entire malodor suppressing composition of the present invention.

[0079] Furthermore, the malodor control composition of the present invention can be blended in an amount commonly used in the industry into, for example, fragrance products, cosmetics, basic cosmetics, finishing cosmetics, hair cosmetics, suntan cosmetics, medicated cosmetics, hair care products, soaps, body cleansers, bath products, laundry detergents, laundry softeners, cleaning agents, kitchen detergents, bleaches, aerosol agents, deodorizers / fragrances, repellents, and miscellaneous goods to enhance their commercial value.

[0080] For example, fragrance products include perfume, eau de parfum, eau de toilette, eau de cologne, etc.; basic cosmetics include facial cleansing cream, vanishing cream, cleansing cream, cold cream, massage cream, milk lotion, lotion, serum, pack, makeup remover, etc.; finishing cosmetics include foundation, lipstick, lip balm, etc.; hair cosmetics include hair tonic, hair liquid, hair spray, etc.; tanning cosmetics include tanning products, sunscreen products, etc.; medicated cosmetics include antiperspirants, aftershave lotions and gels, permanent wave agents, medicated soaps, medicated shampoos, medicated skin cosmetics, etc.

[0081] Examples of hair care products include shampoo, rinse, rinse-in-shampoo, conditioner, treatment, hair pack, etc.; examples of soaps include cosmetic soap, bath soap, etc.; examples of body cleansers include body soap, body shampoo, hand soap, etc.; examples of bath additives include bath additives (bath salts, bath tablets, bath liquid, etc.), foam baths (bubble baths, etc.), bath oils (bath perfumes, bath capsules, etc.), milk baths, bath jellies, bath cubes, etc.

[0082] Detergents include heavy-duty laundry detergents, light-duty laundry detergents, liquid detergents, laundry soaps, compact detergents, powdered soaps, etc.; fabric softeners include softeners, furniture care, etc.; cleaning agents include cleansers, house cleaners, toilet cleaners, bathroom cleaners, glass cleaners, mold removers, drain cleaners, etc.; kitchen detergents include kitchen soaps, kitchen synthetic soaps, dishwashing detergents, etc.; bleaching agents include oxidation bleaches (chlorine bleaches, oxygen bleaches, etc.), reduction bleaches (sulfur bleaches, etc.), optical bleaches, etc.; aerosol agents include spray types, powder sprays, etc.; deodorizing and air freshening agents include solid types, gel types, liquid types, etc.; miscellaneous goods include various forms such as tissue paper and toilet paper.

[0083] When the malodor control composition of the present invention is used in the above-mentioned products, it may be used in its original state, or in a liquid state obtained by dissolving it in, for example, alcohols or polyhydric alcohols such as propylene glycol or glycerin; in a solubilized or dispersed state obtained by solubilizing or dispersing it using a surfactant such as a nonionic surfactant, anionic surfactant, cationic surfactant, amphoteric surfactant, etc.; or in a microcapsulated state obtained by treating it with an encapsulating agent; any form may be selected depending on the purpose.

[0084] Furthermore, the malodor suppressing composition may be used after being stabilized and sustained-released by being enclosed in an enclosure agent such as cyclodextrin. These are appropriately selected and used depending on the form of the final product, such as liquid, solid, powder, gel, mist, aerosol, etc.

[0085] Furthermore, the malodor suppression composition of the present invention is not an odorant by itself, but is used by blending it with a professional fragrance capable of releasing the malodor suppression composition under the conditions of use / application. In this case, the blending amount of the malodor suppression composition is not particularly limited, but is generally in the range of 0.001 to 20.0% by weight based on the entire product.

[0086] 6. Flavor Composition The flavor composition of the present invention contains as an active ingredient a fragrance component that contains an amino acid sequence having at least 80% identity to the amino acid sequence of OR2C1, OR4Q3, or any of these polypeptides, and suppresses the response intensity to at least one olfactory receptor polypeptide selected from the group consisting of polypeptides that are responsive to one or more malodor-causing substances selected from the group consisting of 1-octen-3-one, 1,5-octadien-3-one, 1-octen-3-ol, and 1,5-octadien-3-ol. The flavor composition of the present invention can be blended with known flavors or conventional additives as described below within a range that does not impair the effects of the present invention, i.e., within a quantitative and qualitative range that can reduce the off-flavor of foods and beverages. The flavor composition of the present invention can also be blended with an oral composition within a range that does not impair the effects of the present invention. Other components that can be blended into the flavor composition of the present invention include various synthetic flavors, natural flavors, natural essential oils, plant extracts, etc., such as the natural essential oils, natural flavors, and synthetic flavors described in "Japan Patent Office Official Gazette, Collection of Well-Known and Commonly Used Techniques (Flavors), Part II Food Flavors, pp. 88-131, published on January 14, 2000." The flavor composition of the present invention may contain, as necessary, commonly used solvents such as water and ethanol; and retaining agents such as ethylene glycol, propylene glycol, dipropylene glycol, hexylene glycol, glycerin, triethyl citrate, medium-chain fatty acid triglycerides, and medium-chain fatty acid diglycerides.

[0087] The food and drink of the present invention includes: Beverages such as fruit juice drinks, fruit liquors, dairy drinks, carbonated drinks, soft drinks, and energy drinks; tea drinks or luxury drinks such as green tea, oolong tea, black tea, persimmon leaf tea, chamomile tea, kumazasa tea, mulberry tea, dokudami tea, pu-erh tea, mate tea, rooibos tea, gymnema tea, guava tea, coffee, and cocoa; soups such as Japanese-style soup, Western-style soup, and Chinese-style soup; various instant drinks, etc.; Frozen desserts such as ice cream, sorbets, and popsicles; desserts such as jellies and puddings; Western confectioneries such as cakes, cookies, chocolates, and chewing gum; Japanese confectioneries such as manju, yokan, and uilou; jams; candies; breads; flavor seasonings; various instant foods; various snack foods, etc.; The oral compositions include toothpaste, oral cleansers, mouthwashes, lozenges, chewing gums, etc.; Examples include: EXAMPLES

[0088] The present invention will be described in more detail below by way of examples, but the present invention is not limited to these examples in any way.

[0089] [Example 1] Measurement of olfactory receptor responses to 1-octen-3-one (1) Cloning of olfactory receptor genes The human olfactory receptor gene was obtained by PCR cloning from Human Genomic DNA: Female (Promega) based on the sequence information registered in GenBank. The N-terminal 20 amino acid residues of bovine rhodopsin were inserted into the pME18S vector, and the obtained human olfactory receptor gene was inserted downstream to obtain a human olfactory receptor gene expression vector. (2) Expression of olfactory receptor genes in HEK293T cells 0.05μg of human olfactory receptor gene expression vector, 0.01μg of RTP1S vector, 0.01μg of firefly luciferase vector pGL4.29 (Promega) containing a cAMP response element promoter, and 0.005μg of Renilla luciferase vector pGL4.74 (Promega) containing a thymidine kinase promoter were dissolved in 10μL of Opti-MEM I (Gibco) to prepare a gene solution (for one well). 100μL of HEK293T cells were seeded on a 96-well plate (Biocoat, Corning) at the number of cells that reached confluence after 24 hours, and the gene solution was added to each well by the lipofection method according to the instructions for use of Lipofectamine3000 to transfect the cells, and the cells were cultured at 37℃ under a 5% carbon dioxide atmosphere for 24 hours. (3) Luciferase reporter gene assay After removing the culture medium, 50μL of a sample of odorant prepared in CD293 (Gibco) medium (with 20μM L-glutamine) was added to each well to obtain a concentration for measurement. After 3 hours of stimulation, luciferase activity was measured according to the instructions for the Dual-Luciferase Reporter Assay System (Promega). The response strength of the olfactory receptor polypeptide was expressed as a FoldIncrease value calculated by dividing the luciferase activity generated by odorant stimulation by the luciferase activity generated in a test system without odorants. (4) Identification of olfactory receptor polypeptides that respond to 1-octen-3-one The receptor response to various concentrations of 1-octen-3-one in cells expressing human olfactory receptors OR2C1 and OR4Q3 was measured by luciferase reporter gene assay. The results are shown in Figures 1 and 2, respectively. Both receptors showed a concentration-dependent response to 1-octen-3-one. On the other hand, no response was observed in mock tests using cells that do not express olfactory receptors. In other words, OR2C1 and OR4Q3 were shown to specifically respond to 1-octen-3-one.

[0090] [Example 2] Measurement of olfactory receptor responses to 1,5-octadien-3-one As in Example 1, the receptor response to various concentrations of 1,5-octadien-3-one was measured by luciferase reporter gene assay in cells expressing human olfactory receptors OR2C1 and OR4Q3. The results are shown in Figures 3 and 4, respectively. Both receptors showed a concentration-dependent response to 1,5-octadin-3-one. On the other hand, no response was observed in the mock test. In other words, it was shown that OR2C1 and OR4Q3 specifically responded to 1,5-octadien-3-one.

[0091] [Example 3] Measurement of olfactory receptor responses to 1-octen-3-ol In the same manner as in Example 1, the receptor response to various concentrations of 1-octen-3-ol was measured by luciferase reporter gene assay in cells expressing the human olfactory receptor OR4Q3. The results are shown in Figure 5. OR4Q3 showed a concentration-dependent response to 1-octen-3-ol. On the other hand, no response was observed in the mock test. In other words, it was shown that OR4Q3 specifically responds to 1-octen-3-ol.

[0092] [Example 4] Measurement of olfactory receptor responses to 1,5-octadien-3-ol As in Example 1, the receptor response to various concentrations of 1,5-octadien-3-ol was measured by luciferase reporter gene assay in cells expressing the human olfactory receptor OR4Q3. The results are shown in Figure 6. OR4Q3 showed a concentration-dependent response to 1,5-octadien-3-ol. On the other hand, no response was observed in the mock test. In other words, it was shown that OR4Q3 specifically responds to 1,5-octadien-3-ol.

[0093] [Example 5] Evaluation of the inhibitory effect of odor-suppressing materials on the OR2C1 response to 1-octen-3-one odor As in Example 1, 30 μM 1-octen-3-one was mixed with various concentrations of the malodor suppression material 2-methyl-4-(2,2,3-trimethylcyclopent-3-enyl)but-2-en-1-ol (also known as 2-methyl-4-(2,2,3-trimethyl-3-cyclopentenyl)-2-buten-1-ol) (Hindinol®) and 1-(2,2,6-trimethylcyclohexyl)hexan-3-ol (Dextramber®), and the receptor response was measured in cells expressing OR2C1. The ratio of the FoldIncrease value in the test in which the malodor suppression material was mixed to the FoldIncrease value in the test in which the malodor suppression material was not mixed was calculated. The results are shown in Figures 7 and 8, respectively. 2-Methyl-4-(2,2,3-trimethylcyclopent-3-enyl)but-2-en-1-ol and 1-(2,2,6-trimethylcyclohexyl)hexan-3-ol showed a concentration-dependent effect of reducing the response of OR2C1 to 1-octen-3-one.

[0094] [Example 6] Evaluation of the inhibitory effect of odor-suppressing materials on the OR2C1 response to trans-2-nonenal odor As in Example 5, 60 μM trans-2-nonenal was mixed with various concentrations of the odor-suppressing materials 2-methyl-4-(2,2,3-trimethylcyclopent-3-enyl)but-2-en-1-ol (Hindinol®) and 1-(2,2,6-trimethylcyclohexyl)hexan-3-ol (Dextramber®), and the receptor response was measured in cells expressing OR2C1. The results are shown in Figures 9 and 10, respectively. 2-Methyl-4-(2,2,3-trimethylcyclopent-3-enyl)but-2-en-1-ol and 1-(2,2,6-trimethylcyclohexyl)hexan-3-ol showed the effect of reducing the response of OR2C1 to trans-2-nonenal in a concentration-dependent manner. [Industrial Applicability]

[0095] By using the screening method of the present invention, it is possible to select candidate substances for 1-octene-3-one odor, 1,5-octadien-3-one odor, 1-octene-3-ol odor, and 1,5-octadien-3-ol odor suppressing materials from a large number of test substances. It is expected that the screening method of the present invention can contribute to the development of 1-octene-3-one odor suppressing materials, 1,5-octadien-3-one odor suppressing materials, 1-octene-3-ol odor suppressing materials, and 1,5-octadien-3-ol odor suppressing materials. In addition, it is expected that the malodor suppressing composition of the present invention can contribute to the reduction of unpleasant aging odor and / or armpit odor.

Claims

1. A method for screening an inhibitory material for one or more malodors selected from the group consisting of 1-octen-3-one odor and 1,5-octadien-3-one odor, comprising the steps of: (a) adding a test substance and one or more malodor-causing substances selected from the group consisting of 1-octene-3-one and 1,5-octadien-3-one to at least one olfactory receptor polypeptide selected from the group consisting of human olfactory receptor polypeptide OR2C1 consisting of the amino acid sequence (SEQ ID NO: 2), human olfactory receptor polypeptide OR4Q3 consisting of the amino acid sequence (SEQ ID NO: 4), and (b) a polypeptide comprising an amino acid sequence having at least 90% identity with the amino acid sequence of any of the polypeptides of (a) and showing responsiveness to one or more malodor-causing substances selected from the group consisting of 1-octene-3-one and 1,5-octadien-3-one; Measuring the response of the olfactory receptor polypeptide to one or more malodor-causing substances selected from the group consisting of 1-octen-3-one and 1,5-octadien-3-one; and identifying a test substance that inhibits the response of the olfactory receptor polypeptide based on the measured response; The method includes:

2. The method according to claim 1, wherein the response of the olfactory receptor polypeptide is measured on cells isolated from an organism expressing the olfactory receptor polypeptide, or on cells in which the olfactory receptor polypeptide is artificially expressed by genetic manipulation.

3. The method described in claim 1, wherein the response of the olfactory receptor polypeptide is measured by a reporter gene assay on a cell in which the olfactory receptor polypeptide is artificially expressed by genetic manipulation.

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