JP7084104B2 - Putrefactive urine odor suppressant - Google Patents

Description

The present invention relates to a putrefactive urine odor suppressant.

In recent years, with the improvement of sewerage system, the problem of strong ammonia odor derived from urine in the toilet has been decreasing. The cause of the ammonia odor of urine is the ammonia produced by urease from urea. Ammonia has a high threshold as a malodorous component (it does not smell unless it has a high concentration), so now that excrement is immediately discharged to the outside due to the spread of flush toilets, ammonia derived from urine It is rare that a strong odor is felt.

On the other hand, in recent years, urine odor different from ammonia odor, which is emitted from urine scattered in the toilet and absorbent articles related to absorption of urine such as used diapers, has become a problem. Such urine odor is generated by the action of the enzyme β-glucuronidase on urine. β-glucuronidase is an enzyme that hydrolyzes a compound (glucuronide) in which various alcohols, phenols, amines and the like are conjugated with glucuronic acid, and is contained in bacteria contained in urine.

Patent Documents 1 and 2 describe that when β-glucuronidase produced by a bacterium acts on urine, phenol, p-cresol, 2-methoxy-4-vinylphenol, 4-vinylphenol, 2-methoxy-1,3-benzene It is described that components such as diol, 1,4-benzenediol, 1,3-benzenediol, and indol increase in urine and significantly increase urine odor intensity. Further, Patent Document 3 describes a used diaper with a composition of p-cresol and other phenolic compounds having 6 to 10 carbon atoms, for example, phenol, dimethylphenol, methoxyphenol, methoxyvinylphenol, and dihydroxybenzene. It is described that the urine odor emitted from can be reproduced. It is considered that these components have a high contribution to the urine odor different from the ammonia odor generated by the action of β-glucuronidase, which is emitted from urine scattered in the toilet and used diapers.

On the other hand, some phenolic compounds such as phenol, 2-methoxy-4-vinylphenol, and 4-vinylphenol may be contained in a trace amount in the initial urine immediately after urination (Patent Document 2). Therefore, even urine immediately after urination may emit a urine odor different from the above-mentioned ammonia odor, albeit slightly.

Absorbent articles related to urine absorption include not only diapers for infants and care recipients, but also light incontinence products such as diapers for light incontinence and urine absorbing pads, and sanitary products for women. This includes products that may be absorbed. Especially in recent years, not only diapers for infants and care recipients, but also demand for light incontinence products such as diapers or pads for light incontinence to care for mild incontinence such as elderly people, multiparous women, sickness or the latter is increasing. ing. Since these light incontinence products are used in daily life, prevention of urine odor not only after use but also during wearing is an important issue.

Recently, a method for suppressing the stink by specifically inhibiting the activity of the olfactory receptor for the targeted stink has been provided (for example, Patent Document 4). The mechanism of odor suppression or modulation based on the inhibition of receptor activity by such antagonist substances is called olfactory receptor antagonism. Smell suppression or modulation by the olfactory receptor antagonism can specifically defeat the perception of the targeted stink, unlike deodorization methods by adding other strong odors such as perfumes and air fresheners. It is a preferable deodorizing means because it can be produced and does not cause discomfort due to the strong odor of the fragrance.

Patent Document 4 describes OR1A1, OR2W1, OR5K1, OR5P3, OR8H1, OR10G4 and OR10G7 as olfactory receptors that respond to 2-methoxy-4-vinylphenol, which is a causative agent of β-glucuronidase-treated urine odor. It has been disclosed that by suppressing the response of these olfactory receptors, it is possible to suppress the odor of urine on which β-glucuronidase acts.

Methyldihydrojasmonate (MDJ) is a conventionally used jasmine-based fragrance. Patent Documents 5 to 7 describe that MDJ suppressed the odor of ammonia in urine. Patent Document 8 describes a fragrance for suppressing fecal odor, which contains a jasmine-based fragrance such as MDJ as a keynote fragrance component. However, to date, there have been no reports that MDJ suppressed the odor of urine acted by β-glucuronidase and the odor of 2-methoxy-4-vinylphenol.

International Publication No. 2009/037861 Japanese Patent No. 5719951 Japanese Patent No. 5368696 Japanese Unexamined Patent Publication No. 2015-21167 Japanese Unexamined Patent Publication No. 2002-159565 Japanese Patent Application Laid-Open No. 2002-253651 JP-A-2002-285001. International Publication No. 2013/018805

The present invention relates to an agent for suppressing the odor of β-glucuronidase-acted urine emitted from absorbent articles such as diapers that are being worn or used, and urine that has been scattered in the toilet.

In the present invention, methyldihydrojasmonate (MDJ) suppresses the response of the 2-methoxy-4-vinylphenol receptors OR10G4 and OR10G7, and suppresses the odor caused by 2-methoxy-4-vinylphenol. Furthermore, it was found that β-glucuronidase acting on it suppresses the odor of urine.

Therefore, the present invention provides an agent for suppressing the odor of urine on which β-glucuronidase acts, which contains methyldihydrojasmonate as an active ingredient.
The present invention also provides a 2-methoxy-4-vinylphenol odor suppressant containing methyldihydrojasmonate as an active ingredient.
The present invention also provides a receptor antagonist for the olfactory receptor OR10G7, OR10G4, or an olfactory receptor polypeptide having an equivalent function, which comprises methyldihydrojasmonate as an active ingredient.

According to the present invention, the odor can be suppressed by suppressing the response of the olfactory receptor to 2-methoxy-4-vinylphenol. Further, since 2-methoxy-4-vinylphenol is a causative substance of the urine odor acted by β-glucuronidase, according to the present invention, the urine odor acted by β-glucuronidase, for example, urine scattered in the toilet, is used. It is possible to suppress the odor of urine emitted from absorbent articles such as finished diapers.

Responses of olfactory receptor OR10G4 expressing cells (left) and OR10G7 expressing cells (right) to various concentrations of 2-methoxy-4-vinylphenol. n = 3, error bar = ± SE. Inhibitory effect of MDJ on 2-methoxy-4-vinylphenol response of OR10G4 expressing cells (left) and OR10G7 expressing cells (right). The horizontal axis is the concentration of MDJ, and the vertical axis is the intensity of response to the stimulus of the rotten urine odor substance (2-methoxy-4-vinylphenol) alone as 100%. Represents the response (%) of the expressing cell. n = 3, error bar = ± SE. Sensory evaluation of the 2-methoxy-4-vinylphenol odor suppressing effect of MDJ (n = 9). Control: 2-Methoxy-4-vinylphenol only, Vehicle: Mineral oil only.

As used herein, "suppression of odor by olfactory receptor antagonism" means that by applying the target odor molecule and another molecule together, the other molecule inhibits the receptor activity for the target odor molecule. As a result, it is a means for suppressing the odor recognized by the individual. The suppression of odor by the olfactory receptor antagonism is distinguished from the means of concealing the desired odor by the fragrance of the fragrance, such as the deodorization by the fragrance, even by the means using other molecules as well. An example of the suppression of odor by the olfactory receptor antagonism is the case of using a substance that inhibits the activity of the olfactory receptor such as an antagonist. If a substance that inhibits the activity of an odor molecule that causes a specific odor is applied to the receptor, the activity of the receptor on the odor molecule is suppressed, so that the odor finally perceived by the individual is suppressed. Can be done.

In the present specification, "absorbent articles related to urine absorption" (or simply "absorbent articles") are diapers for infants and care recipients, diapers for light incontinence, light incontinence products such as urine absorbing pads, and women. A product that can be attached to humans or animals and eventually absorb urine, such as feminine hygiene products.

As used herein, the term "rot urine odor" refers to the odor of urine on which β-glucuronidase acts, unless otherwise specified. In the present specification, "urine on which β-glucuronidase has acted" includes those in which β-glucuronidase is positively added and acted on urinating or collected urine, while β-glucuronidase is urinating or collecting urine. Since it is contained in urine and acts on the urine, urine that has been urinated or collected for a long time is also included in urine on which β-glucuronidase has acted. Therefore, specific examples of the "rot urine odor" (that is, the odor of urine on which β-glucuronidase has acted) in the present specification are the odor emitted from urine that has been pissed or aged after urine collection, and scattered and dried urine. Examples include the odor emitted from urine, the odor of urine emitted from an absorbent article to which urine is attached during or after use, such as a diaper left after use. These urines contain 2-methoxy-4-vinylphenol produced by the action of the enzyme β-glucuronidase. 2-Methoxy-4-vinylphenol is the causative agent of the "rot urine odor".

As used herein, the "odor due to 2-methoxy-4-vinylphenol (or 2-methoxy-4-vinylphenol odor)" is the odor brought about by 2-methoxy-4-vinylphenol. 2-Meth-4-vinylphenol is contained in urine that has been pissed or aged for a long time after collection, splattered and dried urine, absorbent articles with urine during or after wearing, or urine on which β-glucuronidase has acted. ing. Further, 2-methoxy-4-vinylphenol may be contained in a trace amount in the initial urine immediately after urination (Patent Document 2). Therefore, even an absorbent article immediately after urination or immediately after attachment of urine during wearing may emit a 2-methoxy-4-vinylphenol odor. Therefore, the suppression of 2-methoxy-4-vinylphenol odor is caused not only by the urine odor emitted from the absorbent article during or after the attachment of urine during use, but also by the attachment of urine during attachment. It may also be effective in suppressing the urine odor emitted from the absorbent article immediately after.

As used herein, the term "olfactory receptor polypeptide" refers to an olfactory receptor or a polypeptide having an equivalent function thereof, and a polypeptide having an equivalent function to an olfactory receptor is the same as the olfactory receptor. , Can be expressed on the cell membrane, activated by the binding of olfactory molecules, and when activated, has the function of increasing the amount of intracellular cAMP by activating adenylate cyclase in conjugation with intracellular Gαs. (Nat. Neurosci., 2004, 5: 263-278).

OR10G7 is an olfactory receptor expressed in human olfactory cells. OR10G7 is registered in GenBank as GI: 52218829. OR10G7 is a protein consisting of the amino acid sequence represented by SEQ ID NO: 2, encoded by the gene having the nucleotide sequence represented by SEQ ID NO: 1. As used herein, the "olfactory receptor polypeptide having the same function as OR10G7" comprises an amino acid sequence having at least 80% identity with the amino acid sequence shown in SEQ ID NO: 2, and 2-methoxy-4. -Vinylphenol An olfactory receptor polypeptide that is responsive to odor.

OR10G4 is an olfactory receptor expressed in human olfactory cells. OR10G4 is registered in GenBank as GI: 523171171. OR10G4 is a protein consisting of the amino acid sequence represented by SEQ ID NO: 4, encoded by the gene having the nucleotide sequence represented by SEQ ID NO: 3. As used herein, the "olfactory receptor polypeptide having the same function as OR10G4" comprises an amino acid sequence having at least 80% identity with the amino acid sequence shown in SEQ ID NO: 4, and 2-methoxy-4. -Vinylphenol An olfactory receptor polypeptide that is responsive to odor.

As used herein, "at least 80% identity" with respect to a nucleotide sequence and an amino acid sequence is 80% or more, preferably 85% or more, more preferably 90% or more, still more preferably 95% or more, still more preferably. 98% or more, more preferably 99% or more identity.

As used herein, the identity of nucleotide and amino acid sequences is calculated by the Lipman-Pearson method (Science, 1985, 227: 435-41). Specifically, the analysis is performed using the homology analysis (Search homology) program of the genetic information processing software Genetyx-Win (Ver. 5.1.; Software development) with Unit size to compare (ktup) set to 2. Is calculated by.

As shown in Examples below, the olfactory receptors OR10G4 and OR10G7 are 2-methoxy-4-vinylphenol receptors that exhibit concentration-dependent activation with respect to 2-methoxy-4-vinylphenol (2-methoxy-4-vinylphenol). Figure 1). Therefore, the substance that suppresses the response of OR10G4 or OR10G7 is an individual 2-methoxy-4-vinylphenol odor or due to the olfactory receptor antagonism based on the suppression of the response of the receptor to 2-methoxy-4-vinylphenol. It is possible to change the perception of the odor of urine on which β-glucuronidase acts and suppress the odor.

Methyl 3-oxo-2-pentyl-1-cyclopentaneacetate (MDJ) is a 2-methoxy-4-vinylphenol of the olfactory receptors OR10G4 and OR10G7, as shown in the examples below. It suppressed the response (FIG. 2), indicating that MDJ is an antagonist to OR10G4, OR10G7, or an olfactory receptor polypeptide having equivalent function. Furthermore, MDJ also suppressed the odor of 2-methoxy-4-vinylphenol in the sensory test (Fig. 3). MDJ, which has been known as a fragrance material, suppresses the 2-methoxy-4-vinylphenol response of the receptor by its antagonistic action against OR10G4 or OR10G7, and has a 2-methoxy-4-vinylphenol odor. It was not known to have a suppressive function.

In one aspect of the invention, MDJ, an antagonist of OR10G4 or OR10G7, can be used as an active ingredient to suppress the odor of 2-methoxy-4-vinylphenol. Furthermore, MDJ can be used as an active ingredient for suppressing the odor of urine acted by β-glucuronidase caused by 2-methoxy-4-vinylphenol. Therefore, in one embodiment, the present invention provides an agent for suppressing the odor of urine on which β-glucuronidase containing MDJ as an active ingredient acts. In another embodiment, the present invention provides an agent for suppressing the odor of 2-methoxy-4-vinylphenol containing MDJ as an active ingredient.

MDJs can be purchased commercially, for example, from Zeon Corporation, Tokyo Chemical Industry Co., Ltd., Kao Corporation, etc. ("Synthetic Perfume Chemistry and Product Knowledge" Supplementary Revised Edition, Indo See Motoichi, The Chemical Daily, published in March 2005). Alternatively, MDJ can produce the same substance as the commercially available product or a composition containing the same substance by chemical synthesis.

In one embodiment, the inhibitor of putrefactive urine odor (odor of urine acted on by β-glucuronidase, the same applies hereinafter) of the present invention contains MDJ as an active ingredient. In one embodiment, the putrefactive urine odor suppressant of the present invention may be essentially composed of MDJ. In one embodiment, the 2-methoxy-4-vinylphenol odor suppressant of the present invention contains MDJ as an active ingredient. In one embodiment, the 2-methoxy-4-vinylphenol odor suppressant of the present invention may be essentially composed of MDJ.

In one embodiment, MDJ may be contained in a compound or composition for suppressing putrefactive urine odor or 2-methoxy-4-vinylphenol odor as an active ingredient for suppressing the odor. Alternatively, MDJ is a compound or composition for producing a suppressor of rotten urine odor or 2-methoxy-4-vinylphenol odor, or for suppressing rotten urine odor or 2-methoxy-4-vinylphenol odor. Can be used for manufacturing. Examples of compounds or compositions for suppressing rotting urine odor or 2-methoxy-4-vinylphenol odor include toilet deodorants or air fresheners, toilet cleaners, underwear or diaper detergents or fabric softeners. Agents and the like can be mentioned.

In one embodiment, MDJs are used for any compound or composition for which suppression of rotting urine odor or 2-methoxy-4-vinylphenol odor is desired, or for rotting urine odor or 2-methoxy-4-vinylphenol odor. It can be used as an active ingredient for suppressing the odor in any environment where the suppression of the odor is desired. Alternatively, MDJ can be used as an active ingredient for suppressing putrefactive urine odor or 2-methoxy-4-vinylphenol odor for the production of a compound or composition in which suppression of the odor is desired. Examples of compounds or compositions for which suppression of rotting urine odor or 2-methoxy-4-vinylphenol odor is desired include absorbent articles (eg, diapers for infants and care recipients, diapers for light incontinence and urinal pads). Light incontinence products such as, sanitary products for women), bed pads, bed mats, sheets, blankets, underwear, toilet bowls, toilet seat sheets, toilet mats, toilet walls or floor materials, etc.

The above-mentioned agent for suppressing putrefactive urine odor or 2-methoxy-4-vinylphenol odor, and compounds or compositions for which suppression of the odor is desired include, in addition to MDJ, other components having a deodorizing effect. It may be contained as appropriate depending on the above. Another component having the deodorizing effect is Cyclohexadec-8-en-1-one (Grobanone®), which is a component that inhibits the enzyme β-glucuronidase that generates 2-methoxy-4-vinylphenol in urine. And so on. Further, the odor suppressant and the compound or composition desired to suppress the odor may contain any component generally added to the deodorant or deodorant.

As a further aspect, the present invention provides a method for suppressing putrefactive urine odor, which comprises a step of allowing putrefactive urine odor and MDJ to coexist. The present invention also provides a method for suppressing 2-methoxy-4-vinylphenol odor, which comprises a step of coexisting 2-methoxy-4-vinylphenol odor and MDJ.

In one embodiment of the method of the present invention, by applying MDJ to a subject in the presence of putrefactive urine odor or 2-methoxy-4-vinylphenol odor, the odor and MDJ coexist. Thereby, the OR10G4 or OR10G7 of the subject binds to its antagonist MDJ, and the response of the olfactory receptor to the 2-methoxy-4-vinylphenol odor is suppressed, so that the olfactory receptor antagonism suppresses the response of the olfactory receptor. , The odor of rotting urine or the odor of 2-methoxy-4-vinylphenol is suppressed.

In the above method of the present invention, the target is a target that requires suppression of rotten urine odor or 2-methoxy-4-vinylphenol odor, preferably rotten urine odor due to olfactory receptor antagonism or 2-methoxy-4- Examples include subjects that require suppression of the odor of vinylphenol. The subject is not particularly limited as long as it is a mammal, but is preferably a human. More specifically, examples of objects in the method of the present invention that require suppression of rotting urine odor or 2-methoxy-4-vinylphenol odor include sick people, absorbent articles (eg, for infants and care recipients). Users of diapers, light incontinence products such as diapers for light incontinence and urine absorbing pads, menstrual products for women), care workers of the sick or elderly, cleaners, people engaged in garbage collection work, etc. ..

In one embodiment of the method of the present invention, MDJ is preliminarily applied in an environment where putrefactive urine odor or 2-methoxy-4-vinylphenol odor may occur. As a result, when the odor is generated in the environment, MDJ is applied to the subject in the presence of the odor, so that the olfactory receptor antagonism causes the odor of rotting urine or 2-methoxy-4-vinylphenol odor. It is suppressed.

In one embodiment of the method of the invention, MDJ is incorporated into a compound or composition containing putrefactive urine odor or 2-methoxy-4-vinylphenol. This reduces the activity of OR10G4 or OR10G7 with respect to 2-methoxy-4-vinylphenol contained in the compound or composition, suppresses the recognition of rotten urine odor or 2-methoxy-4-vinylphenol odor in the subject, and suppresses the recognition of rotten urine odor or 2-methoxy-4-vinylphenol odor. As a result, the odor is suppressed. In yet another embodiment, MDJ is incorporated into a compound or composition that may generate putrefactive urine odor or 2-methoxy-4-vinylphenol. Thereby, MDJ reduces the activity of OR10G4 or OR10G7 with respect to 2-methoxy-4-vinylphenol generated from the compound or composition, and suppresses the recognition of rotten urine odor or 2-methoxy-4-vinylphenol odor in the subject. As a result, the odor is suppressed. Examples of compounds or compositions that can generate rotting urine odor or 2-methoxy-4-vinylphenol include urine, absorbent articles (eg, infant and caregiver diapers, light incontinence diapers and urine). Light incontinence products such as diapers, sanitary products for women), bed pads, bed mats, sheets, blankets, underwear, toilet bowls, toilet seat sheets, toilet mats, toilet walls or floor materials, etc. Therefore, as yet another aspect of the present invention, an absorbent article containing MDJ (for example, diapers for infants and care recipients, light incontinence products such as diapers for light incontinence and urinal pads, sanitary products for women). , Bed pads, bed mats, sheets, blankets, underwear, toilet bowls, toilet seats, toilet mats, toilet walls or flooring, toilet deodorants or fragrances, toilet cleaners, or underwear or diapers Detergents or softeners are provided.

As an exemplary embodiment of the present invention, the following substances, production methods, uses, methods and the like are further disclosed in the present specification. However, the present invention is not limited to these embodiments.

[1] An agent for suppressing the odor of urine on which β-glucuronidase acts, which contains methyldihydrojasmonate as an active ingredient.

[2] A 2-methoxy-4-vinylphenol odor suppressant containing methyldihydrojasmonate as an active ingredient.

[3] Use of methyldihydrojasmonate for suppressing the odor of urine acted by β-glucuronidase.

[4] Use of methyldihydrojasmonate for suppressing 2-methoxy-4-vinylphenol odor.

[5] Use of methyldihydrojasmonate for the production of an agent for suppressing the odor of urine on which β-glucuronidase has acted.

[6] Use of methyldihydrojasmonate for the production of 2-methoxy-4-vinylphenol odor suppressant.

[7] A method for suppressing the odor of urine on which β-glucuronidase acts, which comprises applying methyldihydrojasmonate to a subject in need thereof.

[8] A method for suppressing a 2-methoxy-4-vinylphenol odor, which comprises applying methyldihydrojasmonate to a subject in need thereof.

[9] In the above [1], [3], [5] and [7], the odor of urine acted by β-glucuronidase is the odor emitted from urine that has been pissed or aged after urine collection, and scattered and dried urine. The odor emitted from, or the odor of urine emitted from an absorbent article to which urine is attached during or after wearing.

[10] A receptor antagonist for OR10G7, OR10G4, or an olfactory receptor polypeptide having an equivalent function, which comprises methyldihydrojasmonate as an active ingredient.

[11] Use of methyldihydrojasmonate as a receptor antagonist against OR10G7, OR10G4, or an olfactory receptor polypeptide having an equivalent function thereof.

[12] In the above [10] or [11],
Preferably, OR10G7 is an olfactory receptor polypeptide consisting of the amino acid sequence set forth in SEQ ID NO: 2.
Preferably, the olfactory receptor polypeptide having the same function as OR10G7 comprises an amino acid sequence having at least 80% identity with the amino acid sequence set forth in SEQ ID NO: 2 and with respect to 2-methoxy-4-vinylphenol odor. Responsive olfactory receptor polypeptide
Preferably, OR10G4 is an olfactory receptor polypeptide consisting of the amino acid sequence set forth in SEQ ID NO: 4.
Preferably, the olfactory receptor polypeptide having the same function as OR10G4 comprises an amino acid sequence having at least 80% identity with the amino acid sequence set forth in SEQ ID NO: 4 and with respect to 2-methoxy-4-vinylphenol odor. It is a responsive olfactory receptor polypeptide.

Hereinafter, the present invention will be described in more detail with reference to examples.

The odorous substances and test substances used in this example are shown in Table 1 below.

Example 1 Identification of 2-methoxy-4-vinylphenol receptor 1) Preparation of olfactory receptor-expressing cells Human olfactory receptors are human genometric DNA female (G1521: Promega) based on the sequence information registered in GenBank. Was cloned by the PCR method using the above as a template. The gene amplified by the PCR method was incorporated into the pENTR vector (Invitrogen) according to the manual, and the NotI and AscI sites present on the pENTR vector were used to prepare the NotI and AscI sites downstream of the Flag-Rho tag sequence on the pME18S vector. Recombined into. In addition, a gene encoding human RTP1S, which is a protein that translocates the olfactory receptor protein produced in cultured cells to the cell membrane surface, was incorporated into the EcoRI and XhoI sites of another pME18S vector.
A reaction solution having the composition shown in Table 2 was prepared, allowed to stand in a clean bench for 15 minutes, and then added to each well of a 96-well plate (BD). Then, 100 μL of HEK293 cells (2 × 10 ⁵ cells / cm ² ) were seeded in each well and cultured at 37 ° C. in an incubator holding 5% CO ₂ for 24 hours. As a control, cells (mock) that do not express the olfactory receptor were prepared.

2) Luciferase assay The olfactory receptor expressed in HEK293 cells increases the amount of intracellular cAMP by activating adenylate cyclase in conjugation with intracellular Gαs. For the measurement of the 2-methoxy-4-vinylphenol response of the olfactory receptor in this study, we used the luciferase reporter gene assay, which monitors the increase in intracellular cAMP level as the luminescence value derived from the firefly luciferase gene (fluc2P-CRE-hygro). board. In addition, a gene fused with a sea urchin shiitake luciferase gene (hRluc-CMV) was simultaneously introduced downstream of the CMV promoter and used as an internal standard for correcting errors in gene transfer efficiency and cell number.
The medium was removed from the culture prepared in 1) above, and 75 μL of a solution containing 2-methoxy-4-vinylphenol prepared in a new medium at a predetermined concentration was added. The cells were cultured in a CO ₂ incubator for 4 hours to fully express the luciferase gene in the cells. The activity of luciferase was measured using Dual-Glo ^TM luciferase assay system (Promega) according to the operation manual of the product. The value obtained by dividing the luminescence value derived from firefly luciferase induced by 2-methoxy-4-vinylphenol stimulation by the luminescence value in cells not stimulated with 2-methoxy-4-vinylphenol was calculated as a field increase, and the response was obtained. It was used as an index of strength.

3) Results As a result of measuring the response to 2-methoxy-4-vinylphenol for cells expressing each of the 387 types of olfactory receptors, OR10G4 was used as an olfactory receptor activated against 2-methoxy-4-vinylphenol. And OR10G7 were found. The response of OR10G4 expressing cells and OR10G7 expressing cells to 2-methoxy-4-vinylphenol was concentration-dependent (Fig. 1). From these results, OR10G4 and OR10G7 were identified as 2-methoxy-4-vinylphenol receptors that recognize 2-methoxy-4-vinylphenol.

Example 2 Identification of OR10G4 or OR10G7 antagonist 1) Luciferase assay A luciferase assay using OR10G4 or OR10G7 expressing cells in the same procedure as in Example 1 against the 2-methoxy-4-vinylphenol response of OR10G4 or OR10G7 expressing cells. The effect of the test substance was investigated. The concentration of 2-methoxy-4-vinylphenol was set to 300 μM. The response intensity of the cells expressing the olfactory receptor was calculated as follows. For various stimulus conditions, the value fLuc / hRluc was calculated by dividing the luminescence value derived from firefly luciferase by the luminescence value derived from sea urchin luciferase. The fLuc / hRluc value (X) induced by 2-methoxy-4-vinylphenol monostimulation was divided by the fLuc / hRluc value (Y) in cells into which the same receptor was introduced and not stimulated, and 2- Receptor activity Mold increase (X / Y) by methoxy-4-vinylphenol monostimulation was determined. Similarly, the fLuc / hRluc value (Z) due to stimulation with a mixture of 2-methoxy-4-vinylphenol and the test substance is divided by the fLuc / hRluc value (Y) in unstimulated cells, and the test substance is used. The receptor activity (Z / Y) at the time of addition was determined. The ratio of the receptor activity (Z / Y) when the test substance was added to the receptor activity (X / Y) by stimulation with 2-methoxy-4-vinylphenol alone was calculated by the following formula, and in the presence of the test substance. Response intensity (Receptor%) was determined. Independent experiments were performed 2-3 times, and the average value of each experiment was obtained.
Response (%) = (Z / Y) / (X / Y) x 100

2) Results Methyldihydrojasmonate (MDJ) suppressed the 2-methoxy-4-vinylphenol response of OR10G4 expressing cells and OR10G7 expressing cells in a concentration-dependent manner, and was determined to be an antagonist of OR10G4 and OR10G7 (OR10G4 and OR10G7). Figure 2).

Example 3 2-Methoxy-4-vinylphenol odor suppressing ability by MDJ The 2-methoxy-4-vinylphenol odor suppressing effect by MDJ, which is an OR10G4 and OR10G7 antagonist identified in Example 2, was confirmed by a sensory test.
For 2-methoxy-4-vinylphenol, a 1% (v / v) aqueous solution of distilled water was prepared. MDJ was dissolved in mineral oil (Sigma) to prepare a 0.1% (v / v) solution, which was used as a test substance solution.
Put two cotton balls in a 20 mL glass bottle (Maruem, No. 6), and soak one cotton ball with 30 μL of 2-methoxy-4-vinylphenol aqueous solution and the other cotton ball with 30 μL of the test substance solution. I let you. The glass bottle containing the cotton ball was covered and allowed to stand at 37 ° C. for 1 hour, and then used as a test sample for the test. Only 2-methoxy-4-vinylphenol was prepared as a reference sample (Control), and only mineral oil was prepared as a control sample (Vehicle).

The sensory test was performed by 9 evaluators in a single-blind manner. The test was conducted near the draft to prevent the spread of odors. In order to eliminate the effect of adaptation to odor, the perceived intensity of malodor from the reference sample was confirmed as appropriate during the test, and breaks were taken if necessary. The samples to be evaluated were presented at random, and the 2-methoxy-4-vinylphenol odor suppressing effect of the test substance was evaluated. In the evaluation of the test sample, the following five criteria "2-methoxy-4-vinylphenol odor is 1: not understood, 2: feels weak, 3: is easy to understand, 4: feels strong, 5: is unbearably strong. When "feel" was set and the malodor intensity of the reference sample was 3, the malodor intensity of each test sample was evaluated on a 9-point scale from 1.0 to 5.0 in increments of 0.5.

The evaluation results of the sensory test by each evaluator are shown in FIG. MDJ has all nine subjects respond with a weaker 2-methoxy-4-vinylphenol odor intensity (score less than 3) compared to when 2-methoxy-4-vinylphenol was sniffed alone. It showed a suppressive effect. From the results of this example, it was shown that MDJ, which is an antagonist of OR10G4 and OR10G7, is effective in suppressing the urine odor acted by β-glucuronidase or the 2-methoxy-4-vinylphenol odor.

Claims (2)

A 2-methoxy-4-vinylphenol odor suppressant containing methyldihydrojasmonate as an active ingredient. A receptor antagonist for the olfactory receptor OR10G7, OR10G4, or an olfactory receptor polypeptide having an equivalent function, which comprises methyldihydrojasmonate as an active ingredient.
OR10G7 is an olfactory receptor polypeptide consisting of the amino acid sequence shown in SEQ ID NO: 2.
OR10G4 is an olfactory receptor polypeptide consisting of the amino acid sequence represented by SEQ ID NO: 4.
The olfactory receptor polypeptide having the same function as OR10G7 consists of an amino acid sequence having 90% or more identity with the amino acid sequence shown in SEQ ID NO: 2, and has a responsiveness to 2-methoxy-4-vinylphenol odor. It is an olfactory receptor polypeptide that has
The olfactory receptor polypeptide having the same function as OR10G4 consists of an amino acid sequence having 90% or more identity with the amino acid sequence shown in SEQ ID NO: 4, and has a responsiveness to 2-methoxy-4-vinylphenol odor. It is an olfactory receptor polypeptide that has
Receptor antagonist.

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