JP3635577B1 - Peroxide radical removing agent in gas phase and method for removing peroxide radical in gas phase - Google Patents

Abstract

（式中、ｎは１〜４の整数を、Ｒ¹¹は置換又は無置換の炭素数２〜８のアルケニル基を示す。） 【Task】
A remover for removing peroxide radicals and a method for removing peroxide radicals are disclosed.
[Solution]
A gas peroxide peroxide removing agent containing a compound represented by the following general formula (1) was employed.
General formula (1)
[Chemical 1]

(In the formula, n represents an integer of 1 to 4, and R ¹¹ represents a substituted or unsubstituted alkenyl group having 2 to 8 carbon atoms.)

Description

  The present invention relates to a removal agent and a removal method for removing peroxide radicals in a gas phase.

Technical background

  In recent years, it has been confirmed that free radicals play an important role in the elucidation of molecular mechanisms of various diseases. In particular, lung cancer is thought to be largely related to the factors that cause free radicals.

  Furthermore, among free radicals, peroxide radicals are particularly problematic. That is, the peroxide radical has characteristics different from other reactive oxygen species from the viewpoint of DNA damage. This point is described in Non-Patent Document 1 as follows. That is, peroxide radicals have an extremely long half-life compared to other reactive oxygen species and high chemical selectivity. For example, peroxide radicals react with thymine, which is the base of DNA, and are preferred. In particular, it is described that the methyl group at the 5-position is oxidized. As a result, 5-hydroperoxymethyl-2'-deoxyuridine (5- (hydroperoxymethyl) -2'-deoxyuridine, 5-formyl-2) is highly mutagenic. It is described to produce '-deoxyuridine) and 5- (hydroxymethyl) -2'-deoxyuridine (5- (hydroxymethyl) -2'-deoxyuridine). On the other hand, it is described that the oxidation of the methyl group at the 5-position by a hydroxy radical is a very minor reaction. It is also described that peroxide radicals also cause degradation of phosphodiester that leads to DNA single- and double-strand breaks (Non-Patent Document 1).

  Furthermore, in experiments using a drug-induced pulmonary DNA damage model in mice, peroxidation radicals were not suppressed by SOD or catalase, and peroxygen radicals were detected by ESR analysis. It has also been reported that peroxide radicals play an important role in DNA damage (Non-patent Document 2).

  Under such circumstances, various free radical scavengers have been studied. For example, JP-T-2001-516771 describes the use of L-glutathione, L-selenomethionine, vitamin C, vitamin E and the like for removing free radicals (Patent Document 1).

Chem Res Toxicol. 1997 Feb; 10 (2): 234-41. Biochem Biophys Res Commun. 1990 Apr 16; 168 (1): 58-64. JP-T-2001-516771

As described above, removers that remove free radicals in the gas phase have been studied. Therefore, the inventor examined the effect of the above-mentioned free radical removing agent by using tobacco smoke that is said to contain a large amount of free radicals. However, even when glutathione or vitamin C was used, there was little change in the total amount of free radicals in tobacco smoke. The inventor has considered and investigated the cause in detail, and has come to focus on “difference in free radical species”. That is, conventionally, superoxide (O _2. ) And hydroxyl radical (HO.) Have been widely studied as free radicals. However, in addition to the above, free radicals include carbon-centered radicals, nitric oxide radicals (NO), peroxide radicals (ROO.), And the like. Accordingly, the radicals present in the gas phase are not necessarily rich in superoxide (O _2. ) And hydroxyl radicals (HO.), But may contain a large amount of other types of free radicals. Recognized. Therefore, the inventor of the present application has recognized the necessity of examining the concentration of various free radicals in tobacco smoke.

  On the other hand, conventionally, it has been difficult to examine the amount of free radical components in the gas phase. However, as a result of intensive studies by the inventors this time, the inventors succeeded in measuring the concentration of various free radicals in a gas containing a plurality of types of free radicals. Specifically, an electron spin resonance apparatus (ESR) was used as described later. This time, we used tobacco smoke as the gas containing multiple types of free radicals. Table 1 shows the types and concentrations of various free radicals observed in the smoke in one cigarette.

  As described above, it has been clarified that free radicals contained in tobacco smoke are remarkably rich in peroxide radicals. Traditionally, it was not known at all that tobacco smoke contained peroxide radicals. Furthermore, it is considered that the peroxide radical can be generated through the reaction of superoxide or hydroxy radical. As a result, it was considered that the free radical species contained in other gas phases may have a high ratio of peroxide radicals. Furthermore, as described above, peroxide radicals are greatly involved in DNA damage. Therefore, human and animal DNA continues to be damaged by peroxide radicals present in the air. That is, the inventor recognized the necessity of removing peroxide radicals in the gas phase. Although the presence of peroxide radicals has been conventionally known, the detection method and abundance thereof have not been recognized at all. The present invention is very significant in that it was first achieved by recognizing for the first time the abundance of peroxide radicals. That is, an object of the present invention is to disclose a removing agent for removing peroxide radicals and a method for removing peroxide radicals.

（式中、ｎは１〜４の整数を、Ｒ¹¹は、置換又は無置換の炭素数２〜８のアルケニル基を示す。）； Under such circumstances, as a result of extensive studies on substances that remove peroxide radicals in the gas phase, it has been found that the following compounds achieve the object of the present invention. Specifically, a peroxide radical scavenger in the gas phase containing a compound represented by the following general formula (1).
General formula (1)

(Wherein n represents an integer of 1 to 4, and R ¹¹ represents a substituted or unsubstituted alkenyl group having 2 to 8 carbon atoms);

（式中、Ｒ²¹及びＲ²²は、それぞれ独立に、−Ｈ、−ＯＨ、又は−ＯＣＯＲ²³を示す。Ｒ²³は、置換又は無置換の炭素数１〜７のアルキル基、或いは、置換又は無置換の炭素数２〜８のアルケニル基を示す。）； A peroxidic radical scavenger in a gas phase comprising a compound represented by the following general formula (2).
General formula (2)

(In the formula, R ²¹ and R ²² each independently represent —H, —OH, or —OCOR ^23. R ²³ represents a substituted or unsubstituted alkyl group having 1 to 7 carbon atoms, or a substituted or unsubstituted group. Represents an unsubstituted alkenyl group having 2 to 8 carbon atoms);

（式中、ｎは１〜４の整数を、Ｒ¹¹は、置換又は無置換の炭素数２〜８のアルケニル基を示す。）； A method for removing peroxide radicals in a gas phase, wherein a compound represented by the following general formula (1) is used.
General formula (1)

(Wherein n represents an integer of 1 to 4, and R ¹¹ represents a substituted or unsubstituted alkenyl group having 2 to 8 carbon atoms);

（式中、Ｒ²¹及びＲ²²は、それぞれ独立に、−Ｈ、−ＯＨ、又は−ＯＣＯＲ²³を示す。Ｒ²³は、置換又は無置換の炭素数１〜７のアルキル基、或いは、置換又は無置換の炭素数２〜８のアルケニル基を示す。）；
を採用した。 A method for removing peroxide radicals in a gas phase, wherein a compound represented by the following general formula (2) is used.
General formula (2)

(In the formula, R ²¹ and R ²² each independently represent —H, —OH, or —OCOR ^23. R ²³ represents a substituted or unsubstituted alkyl group having 1 to 7 carbon atoms, or a substituted or unsubstituted group. Represents an unsubstituted alkenyl group having 2 to 8 carbon atoms);
It was adopted.

  By employing the peroxide radical removing agent of the present invention, it has become possible to remove many peroxide radicals in the gas phase. In particular, a remover that removes peroxide radicals in the gas phase has not been studied at all, so that it has become a very innovative invention. It has become possible to effectively suppress the occurrence and deterioration of various diseases caused by peroxide radicals.

  Furthermore, since the peroxide radical removing agent of the present invention can be extracted from plants, it has become environmentally friendly. In other words, it has become possible to obtain the most demanded peroxide radical scavenger in the gas phase that is suitable for health and the environment.

（式中、ｎは１〜４の整数を、Ｒ¹¹は、置換又は無置換の炭素数２〜８のアルケニル基を示す。） The peroxide radical removing agent for removing peroxide radicals in the gas phase employed in the present invention is a compound represented by the following general formula (1).
General formula (1)

(In the formula, n represents an integer of 1 to 4, and R ¹¹ represents a substituted or unsubstituted alkenyl group having 2 to 8 carbon atoms.)

  Here, the position of the OH group may be at any position. For example, when n is 2, the OH group may be in any of ortho, para, and meta. The alkenyl group may be a straight chain or may have a branched chain. Examples of the substituent for the alkenyl group include an OH group and an alkoxy group. More preferably, carbon number of an alkenyl group is 5-7. Moreover, you may have a double bond in any position.

（式中、Ｒ²¹及びＲ²²は、それぞれ独立に、−Ｈ、−ＯＨ、又は−ＯＣＯＲ²³を示す。Ｒ²³は、置換又は無置換の炭素数１〜７のアルキル基、或いは、置換又は無置換の炭素数２〜８のアルケニル基を示す。） More specifically, the peroxide radical removing agent for removing peroxide radicals in the gas phase employed in the present invention is a compound represented by the general formula (2).

(In the formula, R ²¹ and R ²² each independently represent —H, —OH, or —OCOR ^23. R ²³ represents a substituted or unsubstituted alkyl group having 1 to 7 carbon atoms, or a substituted or unsubstituted group. An unsubstituted alkenyl group having 2 to 8 carbon atoms is shown.)

In the general formula (2), more preferably, any of R ²¹ and R ²² is —H, —OH, or —OCOR ²³ . Examples of the substituent for the alkyl group or alkenyl group of R ²³ include an alkyl group and an OH group, and a methyl group is preferable.

Specific examples of the compound represented by the general formula (1) or (2) include one or more compounds selected from the group of sicon, shikonin, alkanine, nasin, and anthocyanin. More specifically, in the general formula (2), examples of R ²³ include substituents as shown in Table 2.

  The compound represented by the general formula (1) or (2) may be artificially synthesized, or may be collected and extracted from a plant or the like. Here, when plant pigments such as shikonin, alkanine, and nasin are employed, as described above, it is also preferable in terms of the environment.

  In the case of adopting a plant-derived material, for example, shikonin and alkanine can be used from hard purple roots or soft purple roots of plants, and nasnin extracted from eggplants can be used. A general method can be adopted for extraction. For example, these plants are added to a solvent such as boiled water, ethanol, methanol, ethyl acetate, acetone, polypropylene glycol (PG), butylene glycol (BG), and allowed to stand for about one week to about one week. Can be extracted. These are obtained as a powder or a concentrated liquid by volatilizing the solvent.

  The amount of the removing agent for removing the peroxide radical is 1 mole or more, preferably 1 to 2 moles of the compound represented by the general formula (1) or (2) with respect to 1 mole of the peroxide radical.

  The peroxide radical removing agent of the present invention can be widely used for removing peroxide radicals in the gas phase. For example, it can be widely used for removal of peroxide radicals contained in exhaust gas emitted from factories and automobiles, removal of peroxide radicals used in plasma treatment, sewage treatment, chemical reaction, etc., and elimination. In addition, when the peroxy radical removing agent is included in a mask, work clothes, or the like, it is provided for the safety of workers using the peroxy radical. Generally, the reaction in the gas phase is very difficult unlike in the liquid phase, and the present invention is very innovative from this viewpoint.

  Hereinafter, an example of a specific embodiment of the present invention will be described. The peroxide radical removing agent of the present invention can be used by including it in a filter. Specifically, a solid phase filter is impregnated with a peroxide radical removing agent. Then, by bringing the compound into contact with a gas containing a peroxide radical, the oxidizing action of the peroxide radical is inhibited, and as a result, the free radical is removed.

  As a method for including the peroxide radical removing compound in the filter, a conventionally widely used method can be employed. In particular, from the viewpoint of filter removal efficiency, it is preferable to employ a method of impregnating the solid phase filter. If the solid phase is impregnated, the peroxide radical removing compound acts evenly on the smoke passing through the filter, which is convenient. In the present invention, the solid phase refers to a solid layer. For example, it is a solid and is essentially a solid, a solidified solid, or an aggregate of particles or powder. Impregnation means a state in which at least a part of the peroxide radical removing compound is present in the solid phase. More preferably, it refers to a state in which the solid phase exists almost uniformly. That is, when a gas containing peroxide radicals passes through the filter, the filter can react with part or all of the peroxide radical removing substance. The amount of the peroxide radical removing compound contained in the filter can be appropriately set in accordance with the amount of the peroxide radical to be removed.

  The material used for the filter of the present invention is not particularly defined as long as it does not depart from the spirit of the present invention. For example, acetate, polypropylene, polyamide, polyvinyl alcohol, polystyrene, cotton, silk, paper, rayon, etc. are widely adopted. Can do. Furthermore, materials generally known in filter technology such as activated carbon can be used in combination.

  The filter of the present invention can be used as a cigarette filter or an air purifier filter. In the case of a tobacco filter, the content of the peroxide radical removing compound is preferably 15 mM to 25 mM per cigarette.

  Moreover, when the peroxide radical removing agent of the present invention is included in the filter, it can be used as an index of the amount of peroxide radical trapped in the gas phase by utilizing the color tone change of the peroxide radical removing agent. Here, as the color tone change, for example, in the case of nasin, it is possible to use the color change from purple to yellow. That is, when tobacco smoke comes into contact with the filter, the color tone of the peroxide radical removing compound changes due to the change in pH. In addition, shikonin or alkanine is initially colored red to magenta, but changes to a stable semiquinone type as the peroxide radical is removed, so that shikonin and alkanine itself are yellow or blue. Therefore, these color tone changes can be used as an index of the removal amount of the peroxide radical removing compound.

(1) Measurement of various free radical contents in tobacco smoke The measurement of the contents of various free radicals in tobacco smoke was carried out by the following method applying a quantitative method for antioxidant substances. That is, after being exposed to a free radical reagent by sucking tobacco smoke for 60 seconds, it was transferred to an ESR measurement aqueous solution cell (Labotec) and measured with an ESR apparatus (JEOL-FA100). As the suction device, a mini pump (DP-5) manufactured by Tokyo Deodorant Co., Ltd. was used. Here, as a free radical scavenging reagent, DMPO (5,5-dimethyl-1-pyrrole-N-oxde) (manufacturer: Doujin Chemical), Fe-MGD (iron-N-Methyl-D-gulcamine-dithiocarbamate) (manufacturer) : Doujin Chemical) and DPPH (1,1-diphenyl-2-picrylhydrazyl) (manufactured by Wako Pure Chemical Industries, Ltd.). Here, DMPO captures superoxide (O _2. ), Hydroxyl radical (HO.), And carbon-centered radical. Fe-MGD captures nitric oxide radicals (NO). DPPH captures carbon-centered radicals and peroxide radicals (ROO.). Here, the measurement conditions are as shown in Table 3.

  A change in signal intensity proportional to the exposure time is observed due to the reaction between the free radical scavenging reagent and various free radicals. Therefore, the free radical concentration was measured from the change in signal intensity. Here, the concentration of the free radical scavenging reagent was measured in multiple stages from 1 μM, and the concentration was gradually increased until the amount of decrease in the free radical scavenging reagent became constant. Then, when the concentration became constant, the amount of free radical scavenging reagent decreased was taken as the amount of free radicals. Here, the hydroxyl radical which did not show a large change in signal intensity at a concentration of 1 μM was set to 1 μM or less. For the carbon-centered radical, since no change in signal intensity was observed, it was set below the detection limit. On the contrary, even if the free radical scavenging reagent reaches a saturated state, those whose trapping amount does not become a steady state are shown as containing free radicals having a concentration higher than the saturated state.

  In addition, in the ESR spectrum when DMPO was used as a capture reagent, the peak of DMPO due to carbon center radicals was not obtained, and therefore the amount of carbon center radicals in tobacco smoke was recognized to be below the detection limit. Therefore, in this experiment, the amount of change in DPPH becomes the amount of peroxide radicals. Peroxide radicals were quantified by creating a calibration curve from the signal intensities of DPPH at various concentrations. Furthermore, when plural types of free radicals are supplemented with one reagent, they are distinguished by the position where the signal appears.

  Here, the details of the measuring method will be described by taking the content of peroxide radical as an example. FIG. 1 shows a schematic diagram of ESR measurement. Firstly, the tip (fire side) 1 of a cigarette (trade name: Seven Star, distributor: Nihon Tobacco) is lit, and the smoke is removed from the DPPH solution 3 while sucking gas from the cigarette mouth 2 for 60 seconds. Moved to. Here, the saturated solution in 25 degreeC ethanol was employ | adopted for DPPH solution. In FIG. 1, the filter 5 is provided, but the filter is not employed in this measurement method. After the transfer, the free radical scavenging reagent solution was immediately transferred to the ESR measurement aqueous solution cell 4 (Labotec), and measurement was performed with an ESR device (JEOL-FA100). The concentrations of various free radical scavenging reagents decreased with the passage of exposure time. In DPPH, when the gas suction amount was adjusted to 1.5 L / min, DPPH was 100% erased in 45 seconds. When the gas suction amount was adjusted to 0.75 L / min, DPPH was erased 100% in 90 seconds. Here, since the saturated solubility of DPPH in ethanol at 25 ° C. is 1 mM or more, it was confirmed that tobacco smoke contained peroxide radicals of 1 mM or more.

(2) Method for producing tobacco filter Shikonin (manufactured by Tokyo Chemical Industry Co., Ltd.) as a peroxide radical removing agent was dissolved in ethanol so that its concentration was 10 mM. Next, a cylindrical acetate filter (manufacturer: Sunny Health) having a diameter of 5 mm and a height of 10 mm was placed in a beaker, and the shikonin solution was added from above. At this time, if necessary, the air was removed by pinching with tweezers. After a few minutes, the filter was removed and allowed to dry naturally in the shade for 1-2 days. The obtained product was used as a tobacco filter. As a comparative example, a filter was prepared in the same manner by replacing shikonin with vitamin C (manufactured by Kanto Chemical) and glutathione (manufactured by Sigma).

(3) Effect of removing peroxide radicals from shikonin filter The shikonin filter, vitamin C filter and glutathione filter obtained in (2) above were prepared, and the amounts of peroxide radical removal were compared. That is, as shown in FIG. 1, the gas is sucked from the cigarette mouth 2 for 60 seconds, the gas is passed through one of the filters 5 and then moved to the DPPH solution 3, and the same as (1) above. Measured. The amount of DPPH after 60 seconds is shown in FIG.

  As shown in FIG. 2, 0.75 mM DPPH remained in the shikonin filter. On the other hand, only 0.02 mM DPPH remained in the vitamin C filter and 0.011 mM DPPH remained in glutathione. That is, the shikonin filter removed peroxide radicals 20 times or more that of other filters. On the other hand, in the vitamin C filter and glutathione filter, peroxide radicals reacted with DPPH, and most of DPPH was erased. That is, it became clear that the peroxidic radicals could hardly be removed with the filter.

(4) Peroxygen radical removal effect of shikonin filter-(2)
The above-mentioned (3) peroxide radical removing effect was further verified. Here, except the following point, it carried out by the method similar to said (3).
1) The concentration of DPPH was precisely 1 mM.
2) As a comparative example, instead of shikonin, one using the same amount of vitamin C (manufactured by Kanto Chemical) and epigallocatechin gallate (manufactured by Sunny Health Co., Ltd.) was employed.
3) The gas suction speed was between 0.70 and 0.75 L / min.

  FIG. 3 shows the result of measuring the radical residual rate by ESR measurement for the DPPH solution into which cigarette smoke was blown 30 seconds and 60 seconds after the cigarette was burned for the shikonin filter. Here, control indicates a case where the cigarette is sucked without igniting. As shown in FIG. 3, it was found that the intensity of the ESR signal derived from DPPH decreases depending on the suction time of tobacco smoke. In addition, when the cigarette was not burned, the ESR signal did not decrease regardless of the time.

  FIG. 4 shows DPPH when cigarette smoke is passed through for 60 seconds through three types of filters and blank filters (without antioxidants) containing shikonin, vitamin C or epigallocatechin gallate as antioxidants. The radical residual ratio was measured. Here, the ratio of the remaining amount of DPPH is expressed in% (molar ratio). In the figure, (1) shows a blank filter, (2) shows a vitamin C filter, (3) shows epigallocatechin gallate, and (4) shows a filter containing shikonin. Only the filter containing shikonin of (4) was clearly found to have a high radical residual rate. From this, it was recognized that shikonin is particularly effective as a substance used in a filter for reducing active oxygen / free radical species in tobacco smoke. The amount of peroxide radicals trapped this time was 0.61 mM.

  The present invention is very significant in that 25% or more peroxide radicals can be trapped from a gas having a peroxide radical concentration of 13.3 μmol / L.

The schematic of the measuring apparatus of the density | concentration of various radicals is shown. The removal effect of the radical removal filter of this invention is shown. The ESR spectrum of DPPH is shown. The removal effect (2) of the radical removal filter of this invention is shown.

Explanation of symbols

DESCRIPTION OF SYMBOLS 1 Cigarette tip side 2 Cigarette mouthpiece 3 DPPH solution 4 ESR measurement aqueous solution cell 5 Shikonin filter

Claims (11)

The removal agent of the peroxide radical (ROO *) in a gaseous phase containing the compound represented by following General formula (1).
General formula (1)

(In general formula (1), n represents an integer of 1 to 4, and R ¹¹ represents a substituted or unsubstituted alkenyl group having 2 to 8 carbon atoms.) The removal agent of the peroxide radical (ROO *) in the gaseous phase of Claim 1 whose compound represented by the said General formula (1) is a compound represented by the following general formula (2).
General formula (2)

(In the general formula (2), R ²¹ and R ²² each independently represent —H, —OH or —OCOR ^23. R ²³ represents a substituted or unsubstituted alkyl group having 1 to 7 carbon atoms, or And represents a substituted or unsubstituted alkenyl group having 2 to 8 carbon atoms.) The compound represented by the above general formula (1) is, shea Konin, alkannin, is at least one selected from the group consisting of Nasunin and anthocyanins, over the gas phase according to claim 1 or 2 Remover of oxidation radical (ROO . ) . A method for removing peroxide radicals (ROO . ) In a gas phase, wherein a compound represented by the following general formula (1) is used.
General formula (1)

(In general formula (1), n represents an integer of 1 to 4, and R ¹¹ represents a substituted or unsubstituted alkenyl group having 2 to 8 carbon atoms.) A compound represented by the following general formula (1), by contact with a gas containing peroxide radical (ROO ·), the gas phase and removing the peroxide radicals in the gas (ROO ·) Of removing peroxide radicals (ROO . ) Therein .
General formula (1)

(In general formula (1), n represents an integer of 1 to 4, and R ¹¹ represents a substituted or unsubstituted alkenyl group having 2 to 8 carbon atoms.) In Claim 4 or 5, the compound represented by the said General formula (1) is a compound represented by the following general formula (2) of the peroxide radical (ROO *) in the gaseous phase characterized by the above-mentioned. Removal method.
General formula (2)

(In General Formula (2), R ²¹ and R ²² each independently represent —H, —OH, or —OCOR ^23. R ²³ represents a substituted or unsubstituted alkyl group having 1 to 7 carbon atoms, Alternatively, it represents a substituted or unsubstituted alkenyl group having 2 to 8 carbon atoms.) Compound represented by the general formula (1) is to Konin, alkannin, is at least one selected from the group consisting of Nasunin and anthocyanins, over the gas phase according to any one of claims 4-6 Removal method of oxidation radical (ROO . ) . In the gas phase according to any one of claims 5 to 7, the compound represented by the general formula (1) is used in an amount of 1 mol or more per 1 mol of the peroxide radical (ROO.) Contained in the gas to be contacted. For removing peroxide radicals (ROO . ) The gas phase according to any one of claims 5 to 7, wherein the compound represented by the general formula (1) is used in an amount of 1 to 2 mol per 1 mol of a peroxide radical (ROO.) Contained in a gas to be contacted. Of removing peroxide radicals (ROO . ) Therein . Comprising the step of an index of free radicals trapped amount of color change of the compound represented by general formula (1), peroxide radical in the gas phase according to any of claims 4 to 9 (ROO ·) Removal method. The compound represented by the said General formula (1) is included in a solid-phase filter, The process which passes the gas containing a peroxide radical (ROO *) through this filter is included in any one of Claims 4-10. A method for removing peroxide radicals (ROO . ) In a gas phase.

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