Oxidative stress inhibitor, oxidative stress inhibitory composition, and oxidative stress inhibitory device

Abstract

To inhibit oxidative stress.SOLUTION: An oxidative stress inhibitor comprises a compound represented by chemical formula (I) in the figure or a salt thereof exclusively as an active ingredient.SELECTED DRAWING: Figure 1

Description

[Technical field]

[0001] The present disclosure relates to an oxidative stress-suppressing agent, a composition for suppressing oxidative stress, and an apparatus for suppressing oxidative stress. [Background technology]

[0002] When active oxygen and free radicals become excessive, they can damage the body and tissues, and even cause various diseases. When cells are subjected to various stresses from the outside world, they can be a problem, especially in nerve cells.

[0003] To combat such active oxygen, low molecular weight antioxidants such as vitamin E and catechin have been introduced, as described in Patent Document 1 (JP Patent Publication No. 2011-184429). DISCLOSURE OF THEINVENTION [Problem to be solved by the invention]

[0004] In addition to the antioxidants mentioned above, new substances having antioxidant properties are desired. [Means for solving the problem]

[0005] The inventors conducted extensive research into phthalide compounds capable of suppressing oxidative stress and discovered that sedanolide, which is contained in the aromatic components of celery, has an oxidative stress suppressing effect. Through further research, the inventors have completed the oxidative stress suppressant, composition for suppressing oxidative stress, and device for suppressing oxidative stress, which will be described below.

[0006] The oxidative stress suppressant according to the first aspect contains only a compound represented by the following chemical formula (I) or a salt thereof as an active ingredient.

[0007] [ka]

[0008] This oxidative stress suppressant suppresses oxidative stress.

[0009] A composition for suppressing oxidative stress according to a second aspect is a composition for suppressing oxidative stress, which comprises, as an active ingredient, only a compound represented by the following chemical formula (I) or a salt thereof:

[0010] [ka]

[0011] This composition for suppressing oxidative stress suppresses oxidative stress.

[0012] The composition for suppressing oxidative stress according to a third aspect is the composition for suppressing oxidative stress according to the first or second aspect, and is a food, cosmetic, or medicine.

[0013] This composition for suppressing oxidative stress makes it possible to enhance the antioxidant defense ability of the living body.

[0014] The composition for suppressing oxidative stress according to the fourth aspect is a composition for suppressing oxidative stress according to any one of the first to third aspects, which is used as a spray from a device selected from the group consisting of an air conditioner, an air purifier, a humidifier, a cosmetic spray device, and a medical spray device.

[0015] This composition for suppressing oxidative stress makes it possible to suppress oxidative stress in the user by operating the device.

[0016] An oxidative stress-suppressing device according to a fifth aspect of the present invention comprises the oxidative stress-suppressing composition according to the second aspect of the present invention and a device capable of spraying the oxidative stress-suppressing composition, the device being any one device selected from the group consisting of an air conditioner, an air purifier, a humidifier, a cosmetic spray device, and a medical spray device, According to this oxidative stress suppression device, it is possible to suppress oxidative stress in a user by operating the device. [Brief description of the drawings]

[0017] [Figure 1] FIG. 1 shows the results of Test 1 in Example 1. [Diagram 2] FIG. 1 is a diagram showing the results of Test 1 in Comparative Example 1. [Diagram 3] FIG. 1 is a diagram showing the results of Test 1 in Comparative Example 2. [Figure 4] FIG. 1 shows calculated values ​​of the area under the curve of fold change values ​​for Example 1 and Comparative Examples 1 and 2. [Diagram 5] This is a photograph of the results of test 2. [Figure 6] FIG. 1 shows the results of Test 3. [Figure 7] FIG. 1 shows the results of Test 4. [Figure 8] 1 is a graph showing the concentration of hydrogen peroxide that is IC25. [Figure 9] 1 is a graph showing cell viability by concentration in Example 1 and Comparative Examples 1 and 2. [Figure 10] FIG. 1 is a schematic diagram of an air conditioning device. [Figure 11] 1 is a schematic functional block diagram of an air conditioning apparatus. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0018] The oxidative stress-suppressing agent, the composition for suppressing oxidative stress, and the device for suppressing oxidative stress will be described in further detail below.

[0019] (1) Oxidative stress inhibitors The oxidative stress inhibitor contains only a compound represented by the following chemical formula (I) or a salt thereof as an active ingredient.

[0020] [ka]

[0021] The above compound has the molecular formula C 12H 18 O 2 It is a tetrahydrophthalide compound, also known as sedanolide.

[0022] Sedanolide is a component that is widely found in plants of the Umbelliferae family, such as celery and cnidium, and has been reported to have antibacterial activity, insecticidal activity, and other activities. However, through screening by the inventors for the oxidative stress inhibitory effect, it has been newly confirmed that it also exhibits activity against oxidative stress.

[0023] The oxidative stress inhibitor is formulated with the above-mentioned sedanolide or a pharma- ceutically acceptable salt thereof. The sedanolide or a pharma- ceutically acceptable salt thereof is commercially available.

[0024] The dosage form of the oxidative stress inhibitor is not particularly limited, and examples thereof include tablets, capsules, pills, granules, powders, liquids, syrups, suspensions, etc. These preparations are prepared by known techniques and may contain acceptable conventional carriers, excipients, binders, stabilizers, etc.

[0025] (2) Composition for suppressing oxidative stress The composition for suppressing oxidative stress contains only a compound represented by the following chemical formula (I) or a salt thereof as an active ingredient.

[0026] [ka]

[0027] The composition for suppressing oxidative stress can be, for example, a food product, a cosmetic product, or a medicine.

[0028] The composition for suppressing oxidative stress may be, but is not limited to, a liquid or powder so as to be treated as a fluid. Such a fluid is preferably used as a spray from a device selected from the group consisting of an air conditioner, an air purifier, a humidifier, a cosmetic spray device, and a medical spray device.

[0029] Known air conditioners, air purifiers, humidifiers, cosmetic spray devices, and medical spray devices can be used, and the oxidative stress-suppressing composition can be sprayed by placing the oxidative stress-suppressing composition in the flow path through which the airflow passes.

[0030] The amount of sedanolide in the composition for suppressing oxidative stress is preferably 50 μM or more from the viewpoint of obtaining a significant effect of suppressing oxidative stress. Even if the amount of sedanolide in the composition for suppressing oxidative stress is 100 μM, no cytotoxicity has been confirmed. The amount of sedanolide in the composition for suppressing oxidative stress is not particularly limited, but may be, for example, 500 μM or less, or 200 μM.

[0031] (3) Oxidative stress suppression device The oxidative stress-suppressing device includes a composition for suppressing oxidative stress and a device capable of spraying the composition for suppressing oxidative stress, the device being selected from the group consisting of an air conditioner, an air purifier, a humidifier, a spray device for cosmetic use, and a spray device for medical use.

[0032] The following description will be given taking as an example an air conditioner 1 as a refrigeration cycle apparatus, which is shown in FIG. 10, which is a schematic configuration diagram, and FIG. 11, which is a schematic functional block configuration diagram.

[0033] The air conditioner 1 is a device that conditions the air in a target space by performing a vapor compression refrigeration cycle. The air conditioner 1 mainly has an outdoor unit 20, an indoor unit 30, a liquid side refrigerant connection pipe 6 and a gas side refrigerant connection pipe 5 that connect the outdoor unit 20 and the indoor unit 30, a remote control (not shown) as an input device and an output device, and a controller 7 that controls the operation of the air conditioner 1.

[0034] In the air conditioner 1, a refrigeration cycle is performed in which the refrigerant sealed in the refrigerant circuit 10 is compressed, cooled or condensed, depressurized, heated or evaporated, and then compressed again. In this embodiment, the refrigerant circuit 10 is filled with a refrigerant for performing a vapor compression refrigeration cycle.

[0035] The outdoor unit 20 is connected to the indoor unit 30 via the liquid side refrigerant connection pipe 6 and the gas side refrigerant connection pipe 5, and constitutes a part of the refrigerant circuit 10. The outdoor unit 20 mainly has a compressor 21, a four-way switching valve 22, an outdoor heat exchanger 23, an outdoor expansion valve 24, an outdoor fan 25, a liquid side shut-off valve 29, and a gas side shut-off valve 28.

[0036] The compressor 21 is a device that compresses a low-pressure refrigerant in the refrigeration cycle until it becomes high pressure. The four-way switching valve 22 can switch between a cooling operation connection state in which the discharge side of the compressor 21 is connected to the outdoor heat exchanger 23 and the suction side of the compressor 21 is connected to the gas side shutoff valve 28, and a heating operation connection state in which the discharge side of the compressor 21 is connected to the gas side shutoff valve 28 and the suction side of the compressor 21 is connected to the outdoor heat exchanger 23, by switching the connection state. The outdoor heat exchanger 23 is a heat exchanger that functions as a condenser of high-pressure refrigerant in the refrigeration cycle during cooling operation and as an evaporator of low-pressure refrigerant in the refrigeration cycle during heating operation. The outdoor fan 25 draws outdoor air into the outdoor unit 20, exchanges heat with the refrigerant in the outdoor heat exchanger 23, and then generates an air flow to be discharged to the outside. The outdoor fan 25 is driven to rotate by an outdoor fan motor. The outdoor expansion valve 24 is provided between the liquid side end of the outdoor heat exchanger 23 and the liquid side shut-off valve 29. The liquid side shut-off valve 29 is a manual valve arranged at the connection portion between the outdoor unit 20 and the liquid side refrigerant connection pipe 6. The gas side shut-off valve 28 is a manual valve arranged at the connection portion between the outdoor unit 20 and the gas side refrigerant connection pipe 5.

[0037] The outdoor unit 20 has an outdoor unit control section 27 that controls the operation of each section constituting the outdoor unit 20. The outdoor unit control section 27 has a microcomputer including a CPU, memory, etc. The outdoor unit control section 27 is connected to the indoor unit control sections 34 of each indoor unit 30 via a communication line, and transmits and receives control signals, etc.

[0038] The outdoor unit 20 is provided with a discharge pressure sensor 61, a discharge temperature sensor 62, a suction pressure sensor 63, a suction temperature sensor 64, an outdoor heat exchanger temperature sensor 65, an outdoor air temperature sensor 66, and the like. Each of these sensors is electrically connected to the outdoor unit control unit 27 and transmits a detection signal to the outdoor unit control unit 27. The discharge pressure sensor 61 detects the pressure of the refrigerant flowing through a discharge pipe connecting the discharge side of the compressor 21 and one of the connection ports of the four-way switching valve 22. The discharge temperature sensor 62 detects the temperature of the refrigerant flowing through the discharge pipe. The suction pressure sensor 63 detects the pressure of the refrigerant flowing through a suction pipe connecting the suction side of the compressor 21 and one of the connection ports of the four-way switching valve 22. The suction temperature sensor 64 detects the temperature of the refrigerant flowing through the suction pipe. The outdoor heat exchanger temperature sensor 65 detects the temperature of the refrigerant flowing through the liquid side outlet of the outdoor heat exchanger 23, which is opposite to the side to which the four-way switching valve 22 is connected. The outdoor air temperature sensor 66 detects the outdoor air temperature before passing through the outdoor heat exchanger 23 .

[0039] The indoor unit 30 is installed on the wall or ceiling of the room, which is the target space. The indoor unit 30 is connected to the outdoor unit 20 via the liquid side refrigerant connection pipe 6 and the gas side refrigerant connection pipe 5, and constitutes a part of the refrigerant circuit 10. The indoor unit 30 has an indoor heat exchanger 31, an indoor fan 32, an indoor unit casing 30a, an air filter 37, and a chemical storage section 36. The indoor unit casing 30a is a housing having an intake port 38 and an outlet port 39. The indoor unit casing 30a accommodates the air filter 37, the indoor heat exchanger 31, the indoor fan 32, and the chemical storage section 36, which are arranged in order from the upstream side in the air flow from the intake port 38 to the outlet port 39. The indoor heat exchanger 31 has a liquid side connected to the liquid side refrigerant connection pipe 6 and a gas side end connected to the gas side refrigerant connection pipe 5. The indoor heat exchanger 31 is a heat exchanger that functions as an evaporator of a low-pressure refrigerant in the refrigeration cycle during cooling operation, and functions as a condenser of a high-pressure refrigerant in the refrigeration cycle during heating operation. The indoor fan 32 draws indoor air into the indoor air casing 30a through the suction port 38, exchanges heat with the refrigerant in the indoor heat exchanger 31, and then generates an air flow for discharging the air, together with the drug contained in the drug containing section 36, from the air outlet 39 to the outside of the indoor unit casing 30a. The indoor fan 32 is driven to rotate by an indoor fan motor. The drug containing section 36 contains the above-mentioned composition for suppressing oxidative stress. The drug containing section 36 has an air vent formed therein, and contains the composition for suppressing oxidative stress in a manner that the composition comes into contact with the air flow formed by the indoor fan 32.

[0040] The indoor unit 30 also has an indoor unit control unit 34 that controls the operation of each part constituting the indoor unit 30. The indoor unit control unit 34 has a microcomputer including a CPU, memory, etc. The indoor unit control unit 34 is connected to the outdoor unit control unit 27 via a communication line, and transmits and receives control signals, etc.

[0041] The indoor unit 30 is provided with an indoor liquid side heat exchanger temperature sensor 71, an indoor air temperature sensor 72, etc. Each of these sensors is electrically connected to the indoor unit control unit 34 and transmits a detection signal to the indoor unit control unit 34. The indoor liquid side heat exchanger temperature sensor 71 detects the temperature of the refrigerant flowing through the liquid side outlet of the indoor heat exchanger 31, which is opposite to the side to which the four-way switching valve 22 is connected. The indoor air temperature sensor 72 detects the temperature of the air in the room before it passes through the indoor heat exchanger 31.

[0042] The controller 7 is configured such that the outdoor unit control section 27 and the indoor unit control section 34 are connected via a communication line, and controls the operation of the air conditioner 1. The controller 7 mainly has a processor such as a CPU (Central Processing Unit) and memories such as ROM and RAM. Note that the various processes and controls performed by the controller 7 are realized by the integrated functioning of each section included in the outdoor unit control section 27 and / or the indoor unit control section 34.

[0043] The operation modes of the air conditioner 1 will be explained below.

[0044] The operation modes include a cooling operation mode and a heating operation mode.

[0045] The controller 7 determines whether the operation mode is a cooling operation mode or a heating operation mode based on an instruction received from a remote control or the like, and executes the determined operation mode.

[0046] In the cooling operation mode, the air conditioning device 1 sets the connection state of the four-way switching valve 22 to a cooling operation connection state in which the discharge side of the compressor 21 is connected to the outdoor heat exchanger 23 while connecting the suction side of the compressor 21 to the gas side shut-off valve 28, and circulates the refrigerant filled in the refrigerant circuit 10 mainly through the compressor 21, the outdoor heat exchanger 23, the outdoor expansion valve 24, and the indoor heat exchanger 31 in that order.

[0047] In the heating operation mode, the air conditioning apparatus 1 sets the connection state of the four-way switching valve 22 to a heating operation connection state in which the discharge side of the compressor 21 is connected to the gas side shut-off valve 28 while connecting the suction side of the compressor 21 to the outdoor heat exchanger 23, and circulates the refrigerant filled in the refrigerant circuit 10 primarily through the compressor 21, the indoor heat exchanger 31, the outdoor expansion valve 24, and the outdoor heat exchanger 23 in that order.

[0048] According to the air conditioner 1 described above, it is possible to supply the composition for suppressing oxidative stress contained in the chemical storage section 36 of the indoor unit 30 to the room together with temperature-conditioned air. This allows the user in the room to be provided with a temperature-regulated environment, and oxidative stress is suppressed. The composition for suppressing oxidative stress contained in the chemical storage section 36 is preferably liquid so that it can be easily diffused with the airflow, and is preferably sprayed into the airflow. The spray from the chemical storage section 36 may be one using a known spray structure. The chemical storage section 36 may be one that is controllable by an operation of a remote control or the like to select between a state in which the composition for suppressing oxidative stress is sprayed from the chemical storage section 36 and a state in which the spray is not performed.

[0049] In the above, an air conditioner 1 has been used as an example of an apparatus capable of spraying the oxidative stress-suppressing composition in an oxidative stress-suppressing device, but other devices that can be used include air purifiers, humidifiers, cosmetic spray devices, and medical spray devices.

[0050] An example of an air purifier is one that includes a casing, a fan, an air filter, and a drug storage section that contains a composition for suppressing oxidative stress, and that sprays the composition for suppressing oxidative stress together with purified air.

[0051] An example of a humidifier is one that includes a casing, a fan, a moisture retention section, and a drug storage section that contains the oxidative stress-suppressing composition, and that sprays the oxidative stress-suppressing composition together with humidified air.

[0052] An example of a cosmetic spray device is one that includes a casing, a fan, and a drug storage section that contains a cosmetic drug and an oxidative stress-suppressing composition, and sprays air containing both the cosmetic drug and the oxidative stress-suppressing composition.

[0053] An example of a medical spray device is one that includes a casing, a fan, and a drug storage section that contains a medical drug and an oxidative stress-suppressing composition, and sprays air containing both the medical drug and the oxidative stress-suppressing composition. EXAMPLES

[0054] The contents of the present disclosure will be specifically described below with reference to examples and comparative examples.

[0055] Using Example 1 and Comparative Examples 1 and 2, which are substances with a history of ingestion, as test substances, screening for substances having an inhibitory effect on oxidative stress was carried out in Tests 1 to 3.

[0056] Example 1 In Example 1, sedanolide was used.

[0057] Comparative Example 1 In Comparative Example 1, butylphthalide represented by the following formula (II) was used.

[0058] [ka]

[0059] Comparative Example 2 In Comparative Example 2, n-butylidenephthalide represented by the following formula (III) was used.

[0060] [ka]

[0061] (Test 1) In Test 1, each of the test substances of Example 1, Comparative Examples 1 and 2 was tested for its ability to activate Nrf2 protein.

[0062] Specifically, a test was conducted to determine whether Nrf2 is activated when each of Example 1, Comparative Examples 1, and 2 is applied to a human hepatoma cell line (HepG2 cells). As the human hepatoma cell line (HepG2 cells), a stable cell line was used that was stably introduced with a reporter vector consisting of an antioxidant response element, a thymidine kinase promoter, and luciferase, and that emits light when Nrf2 is activated. As a result, activation of Nrf2 can be confirmed by the phenomenon of increased luminescence intensity.

[0063] The results of Test 1 in Example 1 are shown in Figure 1. This shows the change over time in the degree of Nrf2 activation (luminescence intensity) for each of the cases where the sedanolide concentration was 0 μM (Control), 13.17 μM, 19.75 μM, 29.63 μM, 44.44 μM, 66.67 μM, and 100 μM.

[0064] The results of Test 1 of Comparative Example 1 are shown in Figure 2. This shows the change over time in the degree of Nrf2 activation (luminescence intensity) for each of the cases where the blended concentration of butylphthalide was 0 μM (Control), 13.17 μM, 19.75 μM, 29.63 μM, 44.44 μM, 66.67 μM, and 100 μM.

[0065] The results of Test 1 of Comparative Example 2 are shown in Figure 3. This shows the change over time in the degree of Nrf2 activation (luminescence intensity) for each of the cases where the blended concentration of n-butylidenephthalide was 0 μM (Control), 13.17 μM, 19.75 μM, 29.63 μM, 44.44 μM, 66.67 μM, and 100 μM.

[0066] FIG. 4 shows the results of calculating the area under the curve of the fold change value after 72 hours for each of the substances in Example 1 and Comparative Examples 1 and 2.

[0067] According to the above Figures 1 to 4, it can be seen that the Nrf2 protein was not activated by butylphthalide of Comparative Example 1 and n-butylidenephthalide of Comparative Example 2, whereas Nrf2 was activated by sedanolide of Example 1.

[0068] (Test 2) In Test 2, it was examined whether the sedanolide of Example 1 promotes the translocation of Nrf2 protein into the nucleus of cells.

[0069] Specifically, we treated a human hepatocellular carcinoma cell line (HepG2 cells) with sedanolide for 4 hours and then used an anti-Nrf2 antibody to quantify the nuclear and cytoplasmic localization of Nrf2 protein to examine whether the translocation of Nrf2 protein into the nucleus was promoted.

[0070] Furthermore, under non-stress conditions and without treatment with the oxidative stress-suppressing composition, Nrf2 is stable in a complex with other proteins, whereas in cells treated with the oxidative stress-suppressing composition, it is believed that Nrf2 dissociates from the complex, promoting the translocation of Nrf2 into the nucleus.

[0071] Figure 5 shows photographs of the results of Test 2.

[0072] In FIG. 5, the upper row shows a control without any compounding, and the lower row shows the one containing sedanolide of Example 1 at a concentration of 100 μM. The two upper and lower images on the left side show the results of quantifying the localization of Nrf2 protein by staining it with an anti-Nrf2 antibody. The two upper and lower images in the center show the cell nuclei stained with 4',6-diamidino-2-phenylindole (DAPI). The two upper and lower images on the left side show the localization of Nrf2 protein in the nucleus by superimposing the above two images. According to these, it can be seen that Nrf2 protein has migrated inside the nucleus in the lower right photograph. Therefore, it was confirmed that the use of sedanolide of Example 1 promotes the migration of Nrf2 protein into the nucleus.

[0073] (Test 3) In Test 3, it was examined whether the sedanolide of Example 1 inhibits the generation of active oxygen when oxidative stress is induced in cells.

[0074] Specifically, a human hepatocellular carcinoma cell line (HepG2 cells) was treated with sedanolide and cultured overnight, after which the medium was replaced with one containing menadione (100 μM), an active oxygen generator, and cultured for an additional 2 hours. The amount of active oxygen generated within the cells was quantified using a dye for detecting active oxygen (CellRox (registered trademark)).

[0075] The results of Test 3 are shown in Figure 6. The relative values ​​of the fluorescence intensity of the reactive oxygen detection dye when the sedanolide concentration was 0 μM, 66 μM, and 100 μM are shown. Note that when the sedanolide concentration was 66 μM and 100 μM, a significant difference of 1% was confirmed.

[0076] From the above, it was confirmed that pretreatment of cells with sedanolide suppressed the amount of active oxygen generated in a sedanolide concentration-dependent manner.

[0077] (Test 4) In Test 4, it was examined whether the sedanolide of Example 1 inhibits cell death caused by oxidative stress.

[0078] Specifically, human hepatocellular carcinoma cell line (HepG2 cells) was treated with sedanolide for 24 hours, cultured overnight, exposed to hydrogen peroxide, and the cell viability after 24 hours was measured.

[0079] The results of Test 4 are shown in Figure 7. This shows the cell viability at different concentrations of hydrogen peroxide when the sedanolide concentration was 0 μM, 25 μM, 50 μM, and 100 μM.

[0080] Based on the above, it was confirmed that cell death caused by hydrogen peroxide was suppressed in a concentration-dependent manner by sedanolide.

[0081] With regard to Test 4, Fig. 8 shows a graph showing the concentration of hydrogen peroxide at which the cell viability becomes 75% (IC25) versus the concentration of sedanolide in Example 1. This also shows that the higher the concentration of sedanolide, the higher the concentration of hydrogen peroxide required for cell death, indicating a concentration-dependent inhibitory effect of sedanolide on cell death.

[0082] FIG. 9 shows the results of examining the cell viability at different concentrations for each of sedanolide of Example 1, butylphthalide of Comparative Example 1, and n-butylidenephthalide of Comparative Example 2.

[0083] (Additional Note) Although the embodiments of the present disclosure have been described above, it will be understood that various changes in form and details can be made without departing from the spirit and scope of the present disclosure described in the claims. [Explanation of symbols]

[0084] 1. Air conditioning equipment [Prior art documents] [Patent documents]

[0085] [Patent Document 1] JP 2011-184429 A

Claims

1. An oxidative stress inhibitor comprising as an active ingredient only a compound represented by the following chemical formula (I) or a salt thereof:

2. A composition for suppressing oxidative stress, comprising as an active ingredient only a compound represented by the following chemical formula (I) or a salt thereof:

3. It is a food, cosmetic, or medicine; The composition for suppressing oxidative stress according to claim 2.

4. The composition is used as a spray from a device selected from the group consisting of an air conditioner, an air purifier, a humidifier, a cosmetic spray device, and a medical spray device. A composition for suppressing oxidative stress according to claim 2 or 3.

5. The composition for suppressing oxidative stress according to claim 2, A device selected from the group consisting of an air conditioner (1), an air purifier, a humidifier, a cosmetic spray device, and a medical spray device, the device being capable of spraying the composition for suppressing oxidative stress; An oxidative stress suppression device equipped with

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