WO2022259864A1 - Olfactory receptor inhibitor and application thereof - Google Patents

Abstract

The present invention provides: an olfactory receptor inhibitor that is practical and suitable for deodorants; and a deodorant and a deodorization method using said inhibitor.　An inhibitor of OR2T11 and the like according to the present invention is at least one selected from the group consisting of α-irone, furfuryl methyl disulfide, furfuryl mercaptan, 4-methoxy-2-methyl-2-butanthiol, geraniol, citral, linalyl acetate, terpinyl acetate, acetyl cedrene, δ-damascone, furfuryl methyl sulfide, thiogeraniol, 8-mercaptomenthone, benzyl mercaptan, β-ionone, α-ionone, α-isomethyl ionone, α-damascone, β-damascenone, and β-damascone.

Description

Olfactory receptor inhibitor and use thereof

TECHNICAL FIELD The present invention relates to inhibitors of olfactory receptors and uses thereof.
This application claims priority based on Japanese Patent Application No. 2021-095342 filed with the Japan Patent Office on June 7, 2021, the content of which is incorporated herein.

Olfaction is recognized by olfactory receptors present in olfactory neurons of the olfactory epithelium responding to odor molecules. It is known that one olfactory receptor can respond to multiple odorant molecules, and that one odorant molecule can bind to and act on multiple olfactory receptors.
It has been proposed to suppress the perception of malodors based on the response of olfactory receptors to odor molecules. For example, Patent Literatures 1 and 2 propose a methyl mercaptan odor inhibitor utilizing desensitization of OR4S2. The methyl mercaptan odor inhibitor functions as an OR4S2 agonist.

JP 2020-10794 A JP 2020-10795 A

The methyl mercaptan odor inhibitors of Patent Documents 1 and 2 desensitize receptors to suppress the odor of methyl mercaptan. Therefore, it is necessary to allow the methyl mercaptan odor inhibitor to act on the olfactory receptor in advance before smelling the odor of a sulfur-containing compound such as the odor of methyl mercaptan.
However, in order for this methyl mercaptan odor inhibitor to act in advance and reliably, the agonist must be present in the space to be deodorized at a higher concentration than the sulfur-containing compound odor. It may not be practical.
Accordingly, the present invention provides an olfactory receptor inhibitor suitable for a practical deodorant; a deodorant using the inhibitor; and a deodorizing method.

The present invention has the following aspects.
[1] OR2T11 and an inhibitor that suppresses the response of at least one or more olfactory receptors selected from the group consisting of polypeptides having functions equivalent to OR2T11; Furyl mercaptan, 4-methoxy-2-methyl-2-butanethiol, geraniol, citral, linalyl acetate, terpinyl acetate, acetylcedrene, δ-damascone, furfuryl methyl sulfide, thiogeraniol, 8-mercaptomenthone, benzyl An inhibitor selected from at least one selected from the group consisting of mercaptan, β-ionone, α-ionone, α-isomethylionone, α-damascone, β-damascenone and β-damascone.
[2] OR2T11 and an inhibitor that suppresses the response of at least one or more olfactory receptors selected from the group consisting of polypeptides having functions equivalent to OR2T11; terpinyl acetate, acetylcedrene, β - ionone, alpha-ionone, alpha-isomethylionone, alpha-irone, furfuryl methyl sulfide, furfuryl methyl disulfide, thiogeraniol, benzyl mercaptan, furfuryl mercaptan, alpha-damascone, delta-damascone, beta-damascone, An inhibitor selected from at least one selected from the group consisting of 8-mercaptomenthone and 4-methoxy-2-methyl-2-butanethiol.
[3] OR2T1 and an inhibitor that suppresses the response of at least one or more olfactory receptors selected from the group consisting of polypeptides having functions equivalent to OR2T1; benzyl acetate, methyl benzoate, linalyl acetate, methyl An inhibitor selected from at least one selected from the group consisting of butyrate, α-damascone, α-ionone, α-isomethylionone, β-damascenone, β-damascone, androstenone and diethylsuccinate.
[4] OR2T6 and an inhibitor that suppresses the response of at least one or more olfactory receptors selected from the group consisting of polypeptides having functions equivalent to OR2T6; methylbenzoate, methylbutyrate, β-damascenone and β-damascone, an olfactory receptor inhibitor.
[5] A deodorant containing an inhibitor according to any one of [1] to [4].
[6] A deodorizing method using the inhibitor of any one of [1] to [4].
[7] Use of the inhibitor according to any one of [1] to [4] in deodorizing bad odors involving responses of olfactory receptors.

According to the present invention, an olfactory receptor inhibitor suitable for a practical deodorant; a deodorant using the inhibitor; and a deodorizing method are provided.

FIG. 3 is a diagram showing the results of measuring the response strength (fold increase) of olfactory receptors to hydrogen sulfide in Examples. FIG. 2 is a diagram showing the results of measuring the fold increase of olfactory receptors to methyl mercaptan in Examples. FIG. 3 shows the results of measuring the response of OR2T11 to methyl mercaptan in Examples. FIG. 3 shows the results of measuring the response of OR2T1 to methyl mercaptan in Examples. FIG. 3 shows the results of measuring the response of OR2T6 to methyl mercaptan in Examples. 1 is a diagram showing an overview of the procedure of sensory test 1. FIG. 4 is a graph showing average values of scoring results of sensory test 1. FIG. 4 is a graph showing average values of scoring results of sensory test 2. FIG.

The following terms used herein have the following meanings.
The term "polypeptide having a function equivalent to that of an olfactory receptor" refers to a polypeptide that can be expressed on the cell membrane and that increases intracellular cAMP levels by binding odor molecules.
"Sulfur-containing compound odor" means an odor produced by a compound containing a sulfur atom in its molecule.
An "agonist" is a compound that binds to an olfactory receptor and activates the response of that olfactory receptor.
An "antagonist" is a compound that binds to an olfactory receptor and inhibits agonist activation of that olfactory receptor.
A “partial agonist” is an agonist and is a compound that acts relatively weakly on an olfactory receptor to which the agonist binds and activates the response of the olfactory receptor.
An "inverse agonist" is a compound that binds to an olfactory receptor and inactivates the response of that olfactory receptor. "Inverse agonists" have a high affinity for inactive receptors, shift the equilibrium in favor of inactive receptors, and suppress the generation of intracellular signals.
In the present specification and claims, "-" indicating a numerical range means that the numerical values before and after it are included as lower and upper limits.
The lower and upper limits of the numerical ranges disclosed herein can be arbitrarily combined to form new numerical ranges.

<Olfactory receptor inhibitor>
An inhibitor according to one embodiment suppresses the response of an olfactory receptor to a sulfur-containing compound.
In one embodiment, the olfactory receptor is selected from the group consisting of OR2T11, OR2T1, OR2T6, a polypeptide having an equivalent function to OR2T11, a polypeptide having an equivalent function to OR2T1, and a polypeptide having an equivalent function to OR2T6. At least one or more.

The sulfur-containing compound is not particularly limited as long as it contains a sulfur element in its molecule. Inorganic sulfur compounds such as, for example, hydrogen sulfide and sulfur dioxide;
2-mercapto-3-methyl-1-butanol, 3-mercapto-2-methyl-1-pentanol, 3-mercapto-2-methyl-1-propanol, 3-mercapto-3-methyl-1-butanol, 3 - sulfanylalkanols such as mercapto-1-hexanol, 3-methyl-3(2-methyldisulfanyl)-butan-1-ol, 3-mercapto-3-methyl-1-hexanol;
sulfanilaldehydes such as 3-mercapto-2-methylpentanal, 1-mercapto-3-pentanone, 2-mercapto-3-pentanone, 3-mercapto-3-pentanone, 4-mercapto-4-methyl-2-pentanone, or ketones;
sulfanyl esters such as 3-methyl-3-mercaptobutyl acetate, 3-methyl-3-mercaptobutyl formate;
methyl mercaptan, ethyl mercaptan, allyl mercaptan, t-butyl mercaptan, 2-methylpropyl mercaptan, 1-methoxyheptane-3-thiol, 4-methoxy-2-methylbutane-2-thiol, 1-propanethiol, 2-propanethiol , 2-sulfanylethanol, 2-(methylthio)-2-propanethiol, 3-mercapto-3-methylbutan-1-ol, etc.;
isothiocyanates such as methyl isothiocyanate, ethyl isothiocyanate, allyl isothiocyanate;
thiomethanes such as methylthiomethane, methyldithiomethane, methyltrithiomethane, ethylthioethane and ethyldithioethane; thiophenes such as thiophene, dithiophene, terthiophene, tetrahydrothiophene, 2,5-dimethylthiophene and 2-acetylthiophene;
thiazoles such as 2-isobutylthiazole, 2-acetylthiazole, 4-ethyl-5-propylthiazole;
thiolanes such as 3,5-dimethyl-1,2,4-trithiolane, 1,2,4-trithiolane, 3-methyl-1,3,5-trithiolane;
thioglycolic acid, dithioglycolic acid;
sulfides such as methyl-2-propenyl disulfide, dimethyl sulfide, dimethyl disulfide, dimethyl trisulfide, diallyl sulfide, diallyl disulfide, diallyl trisulfide, diallyl tetrasulfide, thiirane, (1-methylethyl)-thiirane, diphenyl disulfide;
allicin, alliin, ajoene, lentionine, cysteine, glutathione, methylthiomethane, thioacetic acid, methyl thioacetate;

(Inhibitor of OR2T11)
OR2T11 is an olfactory receptor whose expression in human olfactory receptor cells has been confirmed. OR2T11 is registered in GenBank (NCBI) as Gene ID: 127077. OR2T11 is a polypeptide consisting of the amino acid sequence of SEQ ID NO:1.
In one embodiment, OR2T11 can be replaced with a polypeptide having equivalent function as OR2T11. The amino acid sequence of the polypeptide having a function equivalent to OR2T11 preferably exhibits 80% or more homology, more preferably 85% or more homology, and 90% or more homology with the amino acid sequence of OR2T1. more preferably, 95% or more homology is even more preferred, 98% or more homology is particularly preferred, and 99% or more homology is most preferred.

For example, when the sulfur-containing compound is methyl mercaptan, inhibitors such as OR2T11 are α-ylone, furfurylmethyl disulfide, furfuryl mercaptan, 4-methoxy-2-methyl-2-butanethiol, geraniol, citral, linalyl Acetate, terpinyl acetate, acetylcedrene, δ-damascone, furfuryl methyl sulfide, thiogeraniol, 8-mercaptomenthone, benzyl mercaptan, β-ionone, α-ionone, α-isomethylionone, α-damascone, It is selected from at least one selected from the group consisting of β-damascenone and β-damascone.

When the sulfur-containing compound is methyl mercaptan, α-irone, furfurylmethyl disulfide, furfuryl mercaptan, 4-methoxy-2-methyl-2-butanethiol are partial agonists such as OR2T11.
When the sulfur-containing compound is methyl mercaptan, geraniol, citral, linalyl acetate, terpinyl acetate, acetylcedrene, δ-damascone, furfuryl methyl sulfide, thiogeraniol, 8-mercaptomenthone, benzyl mercaptan, OR2T11, etc. is an antagonist.
When the sulfur-containing compound is methyl mercaptan, β-ionone, α-ionone, α-isomethylionone, α-damascone, β-damascenone, β-damascone are inverse agonists such as OR2T11.

For example, when the sulfur-containing compound is hydrogen sulfide, inhibitors such as OR2T11 include terpinyl acetate, acetylcedrene, β-ionone, α-ionone, α-isomethylionone, α-ylone, furfurylmethyl from the group consisting of sulfide, furfurylmethyl disulfide, thiogeraniol, benzyl mercaptan, furfuryl mercaptan, α-damascone, δ-damascone, β-damascone, 8-mercaptomenthone and 4-methoxy-2-methyl-2-butanethiol It is selected from at least one selected.

When the sulfur-containing compound is hydrogen sulfide, terpinyl acetate, acetylcedrene, β-ionone, α-ionone, α-isomethylionone, α-irone, furfurylmethylsulfide, furfurylmethyldisulfide, thiogeraniol , benzyl mercaptan, furfuryl mercaptan are antagonists such as OR2T11.
When the sulfur-containing compound is hydrogen sulfide, α-damascone, δ-damascone, β-damascone, 8-mercaptomenthone, 4-methoxy-2-methyl-2-butanethiol are inverse agonists such as OR2T11.

(Inhibitor of OR2T1)
OR2T1 is an olfactory receptor whose expression in human olfactory receptor cells has been confirmed. OR2T1 is registered in GenBank (NCBI) as Gene ID: 26696. OR2T1 is a polypeptide consisting of the amino acid sequence of SEQ ID NO:2.
In one embodiment, OR2T1 can be replaced with polypeptides having functional equivalents of OR2T1. The amino acid sequence of a polypeptide having a function equivalent to OR2T1 preferably exhibits 80% or more homology, more preferably 85% or more homology, and 90% or more homology with the amino acid sequence of OR2T1. more preferably, 95% or more homology is even more preferred, 98% or more homology is particularly preferred, and 99% or more homology is most preferred.

For example, when the sulfur-containing compound is hydrogen sulfide, inhibitors such as OR2T1 include benzyl acetate, methyl benzoate, linalyl acetate, methyl butyrate, α-damascone, α-ionone, α-isomethylionone, β-damascenone. , β-damascone, androstenone and diethylsuccinate.

When the sulfur-containing compound is hydrogen sulfide, benzyl acetate, methyl benzoate, linalyl acetate, methyl butyrate, α-damascone, α-ionone, α-isomethylionone, β-damascenone, β-damascone, androstenone, diethyl Succinates are antagonists such as OR2T1.

(Inhibitor of OR2T6)
OR2T6 is an olfactory receptor whose expression in human olfactory receptor cells has been confirmed. OR2T6 is registered in GenBank (NCBI) as Gene ID: 254879. OR2T6 is a polypeptide consisting of the amino acid sequence of SEQ ID NO:3.
In one embodiment, OR2T6 can be replaced with a polypeptide having equivalent function as OR2T6. The amino acid sequence of the polypeptide having a function equivalent to OR2T6 preferably exhibits 80% or more homology, more preferably 85% or more homology, and 90% or more homology with the amino acid sequence of OR2T6. more preferably, 95% or more homology is even more preferred, 98% or more homology is particularly preferred, and 99% or more homology is most preferred.

For example, when the sulfur-containing compound is hydrogen sulfide, the inhibitor such as OR2T6 is selected from at least one selected from the group consisting of methylbenzoate, methylbutyrate, β-damascenone and β-damascone.
When the sulfur-containing compound is hydrogen sulfide, methyl benzoate, methyl butyrate, β-damascenone, β-damascone are antagonists such as OR2T6.

(Effect)
The olfactory receptor inhibitor according to one embodiment described above functions as an olfactory receptor antagonist. Therefore, it is not necessary to act on the olfactory receptor before smelling the odor of a sulfur-containing compound. Also, there is no need to have the antagonist present in the target space at a concentration higher than that of the sulfur-containing compound odor.
Therefore, the olfactory receptor inhibitor according to one embodiment is useful as a deodorant.

<Use of deodorant, deodorant method, inhibitor>
According to one embodiment, there is provided a deodorant comprising an olfactory receptor inhibitor according to one embodiment described above. According to the deodorant according to one embodiment, the olfactory receptor inhibitor binds to the olfactory receptor and can suppress the response of the olfactory receptor to the odor of sulfur-containing compounds. As a result, the perception of the sulfur-containing compound odor is inhibited, and the deodorizing effect is exhibited.

According to one embodiment, there is provided a deodorizing method using the olfactory receptor inhibitor according to the above-described embodiment. According to the deodorizing method according to one embodiment, the olfactory receptor inhibitor binds to the olfactory receptor and can suppress the response of the olfactory receptor to the odor of a sulfur-containing compound. As a result, the perception of the sulfur-containing compound odor is inhibited, and the deodorizing effect is exhibited.

In one embodiment, the deodorant may be in the form of an olfactory receptor antagonist or in the form of a composition. In the case of a composition, the olfactory receptor antagonist is included in the composition as an active ingredient for suppressing sulfur-containing compound odors. The composition may further contain components other than the olfactory receptor antagonist as long as the sulfur-containing compound odor suppressing action of the olfactory receptor antagonist is not impaired. Other components include, for example, deodorant components, deodorant components, aromatic components, and additives.

According to one embodiment, a method for suppressing sulfur-containing compound odors using an olfactory receptor antagonist is provided. In the method for suppressing the odor of sulfur-containing compounds, an olfactory receptor antagonist is applied to a subject (individual) whose perception of the odor of methyl mercaptan is to be suppressed. The application of the olfactory receptor antagonist may be performed before the object to be inhibited is exposed to the sulfur-containing compound as long as the sulfur-containing compound odor inhibiting action of the olfactory receptor antagonist is not impaired. may be after exposure to the sulfur-containing compound, or at the same time as the object to be inhibited is exposed to the sulfur-containing compound.

In one embodiment, a deodorant is applied as an olfactory receptor antagonist prior to exposure of the sulfur-containing compound odor control target to the sulfur-containing compound. The applied olfactory receptor antagonist inhibits the response of the olfactory receptor to be inhibited. As a result, even if the suppression target is exposed to the sulfur-containing compound, the response of the olfactory receptor to the sulfur-containing compound is suppressed, and the perception of the smell of the sulfur-containing compound is suppressed.
The deodorant may be carried by an object to suppress sulfur-containing compound odors, may be left standing in a space where sulfur-containing compound odors may occur, or may be mixed with a substance capable of generating sulfur-containing compound odors. good.

The object (individual) to suppress the sulfur-containing compound odor is not particularly limited. For example, those who want to reduce the odor of permanent agents; those who want to reduce the odor of human and animal feces; those who can engage in excretion treatment at home, hospital wards, nursing homes, etc.; those engaged in inspection, cleaning, and construction of drainage pipes and sewage treatment facilities. The subject to be suppressed is not particularly limited, but humans are preferred.
Spaces and substances that can generate sulfur-containing compound odors include, for example, toilets for humans and animals; excrement treatment in medical facilities and nursing care facilities; paper diapers, sanitary products; permanent agents; beauty salons using permanent agents; , underwear, masks, face shields, clothes such as linens, fabrics, fabrics; laundry detergents, softeners; cosmetics, detergents, external agents such as deodorants, medicines; food, etc.; refrigerators; garbage, etc. waste; equipment for manufacturing products that emit sulfur-containing compound odors. However, application of the deodorant and deodorant method is not limited to these examples.

The present invention will be described in more detail below using examples. However, the present invention is not limited to the description of the following examples.

<Preparation of human olfactory receptor-expressing cells>
(pCI-human olfactory receptor vector, pCI-human RTP1S vector)
Genes encoding human olfactory receptors listed in Tables 1 and 2 were cloned based on the sequence information registered in GenBank. Each gene was cloned by PCR using human genomic DNA Human mixed (G3041: Promega) as a template. Each gene amplified by the PCR method was incorporated into a pCI vector (Promega) according to the product protocol. Specifically, the Rho tag sequence is incorporated using the NheI restriction enzyme site and BamHI restriction enzyme site present on the pCI vector, and the Rho tag sequence is integrated using the downstream MluI restriction enzyme site and NotI restriction enzyme site. An olfactory receptor gene was incorporated downstream. Next, the gene encoding human RTP1S was inserted into the MluI restriction enzyme site and NotI restriction enzyme site of the pCI vector.

(olfactory receptor-expressing cells)
Hana3A cells were cultured in 96-well plates (Corning, BioCoat) to 50% confluence. A reaction solution having the composition shown in Table 3 was prepared, allowed to stand on a clean bench for 15 minutes, and then added to each well of a 96-well plate (Corning BioCoat) in an amount of 50 μL. The cells were cultured for 24 hours in an incubator maintained at 37° C. and 5% CO ₂ atmosphere to prepare Hana3A cells expressing each of the 392 types of human olfactory receptors.

<Sulfur-containing compound>
The following odorants were used as sulfur-containing compounds.
・Methyl mercaptan (2% gas/nitrogen) (Sogo Sangyo Co., Ltd.)
・Hydrogen sulfide (1.5% gas/nitrogen) (Sogo Sangyo Co., Ltd.)

<Glo Sensor Assay>
A Glo Sensor assay was performed to measure the responses of the olfactory receptors. Olfactory receptors expressed in Hana3A cells increase the amount of intracellular cAMP by coupling with intracellular Gas and activating adenylate cyclase. An increase in the amount of intracellular cAMP was measured as a luminescence value derived from the firefly luciferase gene, and responses of olfactory receptors were measured.
Luciferase activity was measured using Glo Sensor cAMP Reagent (Promega) according to the product protocol. For various stimulation conditions, the luciferase-derived luminescence value before odor stimulation was divided by the luciferase-derived luminescence value after odor stimulation to calculate (luminescence value after stimulation)/(luminescence value before stimulation). . (Luminescence value after stimulation)/(Luminescence value before stimulation) induced by odorant stimulation was taken as the measured value of response intensity.

<Search for olfactory receptors that respond to sulfur-containing compounds>
(Identification of OR2T11, OR2T1 and OR2T6)
The medium was removed from the culture of olfactory receptor-expressing cells, and 25 μL of Glo Sensor cAMP Reagent diluted with HBSS buffer containing 10 mM HEPES was added to each well of a 96-well plate. After culturing the cells for 2.5 to 3 hours in a light-shielded environment to introduce the cAMP Reagent into the cells, a 96-well plate and an air circulation fan were placed in a 5 L Flex sampler bag and filled with pure air. Finally, methyl mercaptan gas or hydrogen sulfide gas as malodorous molecules was injected with a syringe so that the gas phase final concentration reached a predetermined value. The olfactory receptor-expressing cells were contacted with the malodorous molecules for 10 minutes, and then the Glo Sensor assay was performed to measure the fold increase of the olfactory receptors to the malodorous molecules. The results are shown in FIGS.

The results are expressed as the relative response intensity to odor stimuli in each receptor-expressing cell in the presence of copper ions, where the response intensity in the absence of odor stimulus is set to 1 (Figs. 1 and 2).
As a result of measuring the responses to malodorous molecules in the presence of copper ions, OR2T11 was identified as the olfactory receptor that showed the highest responsiveness to methylmercaptan. Another olfactory receptor responsive to methylmercaptan was identified as OR2T1. OR2T11 was identified as the olfactory receptor with the highest responsiveness to hydrogen sulfide. In addition, OR2T1 and OR2T6 were identified as olfactory receptors that showed responsiveness to hydrogen sulfide.

(Concentration dependence of responses of OR2T11, OR2T1, and OR2T6 to sulfur-containing compounds)
The responses of OR2T11, OR2T1, OR2T6 to different concentrations of methyl mercaptan were measured. Results are shown in FIGS.
As a result, OR2T11 and OR2T1 exhibited a methylmercaptan concentration-dependent response, confirming that they are methylmercaptan receptors. Among them, OR2T11 was found to be the receptor with the highest responsiveness and particularly high sensitivity to methyl mercaptan (Figs. 3, 4 and 5).

<Test substance>
As test substances (Compounds), 105 kinds of compounds were diluted with Glo Sensor Buffer and prepared to 200 μM (final concentration 100 μM).

<Inhibitor of OR2T11>
The medium was removed from the OR2T11-expressing cell culture, and 25 μL of Glo Sensor cAMP Reagent diluted with HBSS buffer containing 10 mM HEPES was added to each well of a 96-well plate. The cells were cultured for 2.5-3 hours to introduce the cAMP Reagent into the cells. After that, Glo Sensor assay was performed to measure the response intensity of the olfactory receptor before adding the test substance to obtain reference data. Next, 25 μL of the test substance was added to each well of the 96-well plate, Glo Sensor assay was performed 10 minutes later, and the response strength (fold increase) of the olfactory receptor to the test substance was measured to obtain test data.
After that, the 96-well plate and a fan for air circulation were placed in a 5 L Flex sampler bag and filled with pure air. Finally, methyl mercaptan as a malodorous molecule was injected with a syringe so that the gas phase final concentration reached a predetermined value. The olfactory receptor-expressing cells were contacted with the malodorous molecules for 10 minutes, and then the Glo Sensor assay was performed to measure the fold increase of the olfactory receptors to the malodorous molecules.
Response strength measurements identified inhibitors of OR2T11. Table 4 shows the measurement results of the response intensity of inhibitors of OR2T11.

As shown in Table 4, these inhibitors suppressed the response of OR2T11 to methyl mercaptan.

Next, the sulfur-containing compound was changed to hydrogen sulfide, and an inhibitor of OR2T11 was similarly identified. Table 5 shows the measurement results of the response intensity.

As shown in Table 5, these inhibitors suppressed the response of OR2T11 to methyl mercaptan.

<Inhibitor of OR2T1>
Inhibitors of OR2T1 were identified in the same manner, except that cultures of OR2T1-expressing cells were used and hydrogen sulfide was used as the sulfur-containing compound. Table 6 shows the measurement results of the response intensity.

As shown in Table 6, these inhibitors suppressed the response of OR2T1 to hydrogen sulfide.

<Inhibitor of OR2T6>
Inhibitors of OR2T6 were identified in the same manner, except that cultures of OR2T6-expressing cells were used and hydrogen sulfide was used as the sulfur-containing compound. Table 7 shows the measurement results of the response intensity.

As shown in Table 7, these inhibitors suppressed the response of OR2T1 to hydrogen sulfide.

<Sensory test 1>
(test sample)
The following test samples were used to test the deodorizing effect of methyl mercaptan odor.
・β-damascone ・δ-damascone ・Iso E super (synthetic fragrance, Tetramethyl Acetylactahydronaphthalenes)

(contents of the test)
Sensory test 1 was conducted by 12 evaluators. First, 0.5 mL of the test sample is impregnated into a filter paper, sealed in a 10 L bag together with a fan, left at room temperature for 1 hour or more to prepare saturated scented air, and the saturated scented air is transferred to 3 L sachet 1. changed. On the other hand, another sachet 2 with a volume of 3 L was prepared and filled with odorless air. After that, 0.5 mL of gaseous methyl mercaptan (2% gas/nitrogen) was injected into each of sachet 1 and 2 to prepare a bag of "odorless air + bad smell" and a bag of "fragrant air + bad smell".
As shown in FIG. 6, the evaluators were simultaneously presented with the odor of methyl mercaptan and the odor of the test sample in the order of δ-damascone, Iso E super, and β-damascone. Each evaluator scored the two items of "comfort/discomfort" and "offensive odor intensity" according to the following criteria.
FIG. 7 and Table 8 show the average values of the scoring results for 12 people.

"Comfort/Discomfort"
Evaluation was made on a 9-point scale from +4 to -4 in 1-point increments according to the following evaluation criteria.
+4: extremely comfortable;
+3: very comfortable;
+2: comfortable;
+1: moderately comfortable;
0: neither comfortable nor uncomfortable;
-1: slightly uncomfortable;
-2: uncomfortable;
-3: very uncomfortable;
-4: Extremely unpleasant.

"Foul Odor Strength"
Evaluation was made on a 6-point scale from +5 points to 0 points according to the following evaluation criteria.
+5: strong;
+4: strong;
+3: easily perceptible;
+2: weak;
+1: barely perceptible;
0: Odorless.

In this example, the standard of sensory deodorant test stipulated by the Aroma Deodorant and Deodorant Council was adopted. The Aromatic Deodorant and Deodorant Association considers a case where the odor intensity or the pleasantness/discomfort level is reduced by 1 or more to be "deodorant effective".
From the results of sensory test 1, β-damascone and δ-damascone showed a superior deodorizing effect than Iso E super. From this, β-damascone and δ-damascone were expected to be useful as deodorants for methyl mercaptan odor. In addition, β-damascone was expected to exhibit deodorizing performance superior to that of δ-damascone.

<Sensory test 2>
(test sample)
The following test samples were used.
・Paraffin (blank)
・β-ionone ・Iso E super (synthetic fragrance, Tetramethyl Acetylactahydronaphthalenes)

(contents of the test)
Sensory test 2 was conducted by 7 evaluators. First, a cotton ball impregnated with 500 μL of the test sample was sealed in a brown bottle with a volume of 15 mL. A 3 L sachet 3 was also filled with odorless air and 1.5 mL of methyl mercaptan (2% gas/nitrogen) was added.
Each evaluator was asked to deeply inhale the test sample 10 times at the mouth of a brown bottle filled with paraffin. Immediately after that, the subjects were asked to smell the odor inside the sachet 3, answer the intensity of the odor, and evaluate it on a scale of 6 according to the same scoring criteria as in the sensory test 1.
Next to paraffin, Iso E super and β-ionone were similarly evaluated in this order for malodor intensity by each evaluator. Since the duration of inhibitory effects by antagonists was unknown, paraffin, Iso E super, and β-ionone were evaluated in that order.
Here, considering the influence of psychological bias, three sachet 3 with the same concentration were prepared and a different sachet was used after each sample.
FIG. 8 and Table 9 show the average values of the scoring results for the seven persons.

From the results of sensory test 2, β-ionone was expected to be useful as a deodorant for the smell of methyl mercaptan.

According to the present invention, an olfactory receptor inhibitor suitable for a practical deodorant; and a practical sulfur-containing compound odor deodorant can be provided.

Claims (7)

1. An inhibitor that suppresses the response of at least one or more olfactory receptors selected from the group consisting of OR2T11 and polypeptides having functions equivalent to OR2T11,
   α-Iron, furfuryl methyl disulfide, furfuryl mercaptan, 4-methoxy-2-methyl-2-butanethiol, geraniol, citral, linalyl acetate, terpinyl acetate, acetylcedrene, δ-damascone, furfuryl methyl sulfide , thiogeraniol, 8-mercaptomenthone, benzylmercaptan, β-ionone, α-ionone, α-isomethylionone, α-damascone, β-damascone and β-damascone selected from at least one selected from the group consisting of , inhibitor.
2. An inhibitor that suppresses the response of at least one or more olfactory receptors selected from the group consisting of OR2T11 and polypeptides having functions equivalent to OR2T11,
   terpinyl acetate, acetylcedrene, β-ionone, α-ionone, α-isomethylionone, α-irone, furfuryl methyl sulfide, furfuryl methyl disulfide, thiogeraniol, benzyl mercaptan, furfuryl mercaptan, α- An inhibitor selected from at least one selected from the group consisting of damascone, δ-damascone, β-damascone, 8-mercaptomenthone and 4-methoxy-2-methyl-2-butanethiol.
3. An inhibitor that suppresses the response of at least one or more olfactory receptors selected from the group consisting of OR2T1 and polypeptides having functions equivalent to OR2T1,
   at least one selected from the group consisting of benzyl acetate, methyl benzoate, linalyl acetate, methyl butyrate, α-damascone, α-ionone, α-isomethylionone, β-damascenone, β-damascone, androstenone and diethylsuccinate An inhibitor selected from more than one species.
4. An inhibitor that suppresses the response of at least one or more olfactory receptors selected from the group consisting of OR2T6 and polypeptides having functions equivalent to OR2T6,
   An olfactory receptor inhibitor selected from at least one selected from the group consisting of methylbenzoate, methylbutyrate, β-damascenone and β-damascone.
5. A deodorant containing the inhibitor according to any one of claims 1 to 4.
6. A deodorizing method using the inhibitor according to any one of claims 1 to 4.
7. Use of the inhibitor according to any one of claims 1 to 4 in deodorizing bad odors in which responses of olfactory receptors are involved.

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