JP2006089661A - Antioxidant - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide an antioxidant capable of preventing various diseases and aging, to provide a pharmaceutical composition, to provide a quasi-drug composition, to provide a cosmetic, and to provide a health food. <P>SOLUTION: This antioxidant contains at least any one of xanthone derivatives expressed by specified five formulae (formulae (2) to (5) are not shown), such as a derivative expressed by formula (1). <P>COPYRIGHT: (C)2006,JPO&NCIPI

Description

  The present invention relates to an antioxidant, a pharmaceutical composition, a quasi-drug composition, a cosmetic, and a health food.

  In the mitochondrial or microsomal electron transport system in the cell, some electrons leak from the system and cause an oxidation-reduction reaction cycle of transition metals such as adjacent iron, reducing oxygen molecules and generating superoxide anions. It continues to produce various active oxygens (hydrogen peroxide, hydroxy radicals) (see Non-Patent Document 1, for example).

These radicals induce a chain peroxidation reaction of lipids. Lipid peroxidation proceeds in a chain, and lipid peroxide destroys membrane lipids one after another, causing problems such as various diseases, carcinogenesis and aging.
Tokyo Metropolitan Geriatric Research and Welfare Foundation Tokyo Metropolitan Institute of Gerontology, “Mitochondrial Genes and Longevity”, [online], [searched on 26 November 2003], Internet <URL: http: //www.tmig .or.jp / J_TMIG / kouenkai / koza / 62koza_3.html>

  Therefore, the present invention is intended to provide an antioxidant, a pharmaceutical composition, a quasi-drug composition, a cosmetic, and a health food that can prevent various diseases and aging.

In order to solve the above problems, the present invention takes the following technical means.
(1) The antioxidant of this invention contains at least one of the xanthone derivatives represented by the following general formulas [Chemical Formula 1], [Chemical Formula 2], [Chemical Formula 3], [Chemical Formula 4], and [Chemical Formula 5]. It is characterized by doing.

この抗酸化剤は前記一般式〔化１〕、〔化２〕、〔化３〕、〔化４〕、〔化５〕で示されるキサントン誘導体のうち少なくともいずれかを含有するものであって、抗酸化作用を有するものであり、様々な疾病や老化を予防する内用剤や外用剤として用いることができる。

This antioxidant contains at least one of the xanthone derivatives represented by the general formulas [Chemical Formula 1], [Chemical Formula 2], [Chemical Formula 3], [Chemical Formula 4], and [Chemical Formula 5], It has an antioxidant action and can be used as an internal preparation or an external preparation for preventing various diseases and aging.

(2) The pharmaceutical composition of the present invention contains at least one of the xanthone derivatives represented by the general formulas [Chemical Formula 1], [Chemical Formula 2], [Chemical Formula 3], [Chemical Formula 4], and [Chemical Formula 5]. It has an antioxidant effect.

  This pharmaceutical composition contains at least one of the xanthone derivatives represented by the above general formulas [Chemical Formula 1], [Chemical Formula 2], [Chemical Formula 3], [Chemical Formula 4], and [Chemical Formula 5], It has an antioxidant action and can be used as an internal preparation or an external preparation for preventing various diseases and aging.

(3) The quasi-drug composition of the present invention comprises at least one of the xanthone derivatives represented by the above general formulas [Chemical Formula 1], [Chemical Formula 2], [Chemical Formula 3], [Chemical Formula 4], and [Chemical Formula 5]. It is characterized by having an antioxidant effect.

  This quasi-drug composition contains at least one of the xanthone derivatives represented by the above general formulas [Chemical Formula 1], [Chemical Formula 2], [Chemical Formula 3], [Chemical Formula 4], and [Chemical Formula 5]. Thus, it has an antioxidant action, and can be used as an internal preparation or an external preparation for preventing various diseases and aging.

(4) The cosmetic of the present invention contains at least one of the xanthone derivatives represented by the above general formulas [Chemical Formula 1], [Chemical Formula 2], [Chemical Formula 3], [Chemical Formula 4], and [Chemical Formula 5]. It has an oxidizing action.

  This cosmetic product contains at least one of the xanthone derivatives represented by the above general formulas [Chemical Formula 1], [Chemical Formula 2], [Chemical Formula 3], [Chemical Formula 4], and [Chemical Formula 5]. It has an action and can be used as an external preparation for preventing various diseases and aging.

(5) The health food of this invention contains at least one of the xanthone derivatives represented by the above general formulas [Chemical Formula 1], [Chemical Formula 2], [Chemical Formula 3], [Chemical Formula 4], and [Chemical Formula 5]. It has an antioxidant effect.

  This health food contains at least one of the xanthone derivatives represented by the above general formulas [Chemical Formula 1], [Chemical Formula 2], [Chemical Formula 3], [Chemical Formula 4], and [Chemical Formula 5]. It has an oxidizing action and can be used as an internal preparation for preventing various diseases and aging.

  The present invention is configured as described above and has the following effects.

  Antioxidants, pharmaceutical compositions, quasi-drug compositions, cosmetics, and health foods that can prevent various diseases and aging can be provided.

  The antioxidant of this embodiment contains at least one of the xanthone derivatives represented by the following general formulas [Chemical Formula 1], [Chemical Formula 2], [Chemical Formula 3], [Chemical Formula 4], and [Chemical Formula 5].

Calopyllum inophyllum根皮のアセトン抽出物を展開溶媒としてn-hexane-EtOAc systemを用い、Si gel CCを行うことにより、7つの部分に粗分画を行った。そのn-hexane-EtOAc＝3：1溶出画分より得られた粗結晶をn-hexane-acetoneで再結晶し、前記一般式〔化１〕に示されるキサントン誘導体（Caloxanthone A）を得た。

Using an acetone extract of Calopyllum inophyllum root bark as an eluent, n-hexane-EtOAc system was used to perform Si gel CC, thereby roughly fractionating seven portions. The crude crystals obtained from the fraction eluted with n-hexane-EtOAc = 3: 1 were recrystallized with n-hexane-acetone to obtain a xanthone derivative (Caloxanthone A) represented by the above general formula [Chemical Formula 1].

Calopyllum inophyllum根皮のアセトン抽出物を展開溶媒としてn-hexane-EtOAc systemを用い、Si gel CCを行うことにより、7つの部分に粗分画を行った。そのn-hexane-EtOAc＝2：1溶出画分より得られた粗結晶をn-hexane-acetoneで再結晶し、前記一般式〔化２〕に示されるキサントン誘導体（macluraxanthone）を得た。

Using an acetone extract of Calopyllum inophyllum root bark as an eluent, n-hexane-EtOAc system was used to perform Si gel CC, thereby roughly fractionating seven portions. The crude crystals obtained from the fraction eluted with n-hexane-EtOAc = 2: 1 were recrystallized with n-hexane-acetone to obtain a xanthone derivative (macluraxanthone) represented by the above general formula [Chemical Formula 2].

Calopyllum inophyllum根材のn-ヘキサン抽出物より析出した粗結晶を濾別した後、濾液の濃縮物についてVLCを行い、n-hexane-EtOAc systemで溶出し、6つの分画が得られた。n-hexane-EtOAc＝10：1溶出画分をn-hexaneより再結晶し、前記一般式〔化３〕に示されるキサントン誘導体（6-deoxyjacareubin（antiinflammation））を得た。

After the crude crystals precipitated from the n-hexane extract of Calopyllum inophyllum rootstock were filtered off, the filtrate concentrate was subjected to VLC and eluted with n-hexane-EtOAc system to obtain 6 fractions. The fraction eluted with n-hexane-EtOAc = 10: 1 was recrystallized from n-hexane to obtain a xanthone derivative (6-deoxyjacareubin (antiinflammation)) represented by the above general formula [Chemical Formula 3].

Calopyllum austroindicum心材を乾燥、粉砕し、アセトンを用いて抽出を行った。溶媒を減圧下除去し、その残査を、Si gel CC（benzene-acetone system）に付し、11の分画を得た。そのうち、3：1の分画をさらにVLC（CHCl3-MeOH system）に付し、5：1溶出画分から前記一般式〔化４〕に示されるキサントン誘導体を得た。

Calopyllum austroindicum heartwood was dried, ground, and extracted with acetone. The solvent was removed under reduced pressure, and the residue was subjected to Si gel CC (benzene-acetone system) to obtain 11 fractions. Among these, the 3: 1 fraction was further subjected to VLC (CHCl3-MeOH system), and the xanthone derivative represented by the above general formula [Chemical Formula 4] was obtained from the 5: 1 eluted fraction.

Calopyllum austroindicum心材を乾燥、粉砕し、ベンゼンを用いて抽出を行った。溶媒を減圧下除去し、その残査を、VLC（n-hexane-EtOAc system）に付し、9つの分画を得た。そのうち、5：1溶出分画から前記一般式〔化５〕に示されるキサントン誘導体（Jacareubin）を、再結晶を行うことによりを得た。

Calopyllum austroindicum heartwood was dried, ground, and extracted with benzene. The solvent was removed under reduced pressure, and the residue was subjected to VLC (n-hexane-EtOAc system) to obtain 9 fractions. Among them, the xanthone derivative (Jacareubin) represented by the general formula [Chemical Formula 5] was obtained from the 5: 1 elution fraction by recrystallization.

  This antioxidant contains at least one of the xanthone derivatives represented by the general formulas [Chemical Formula 1], [Chemical Formula 2], [Chemical Formula 3], [Chemical Formula 4], and [Chemical Formula 5], It has an anti-oxidant action and can be used as an internal preparation or an external preparation for preventing various diseases and aging, and can be applied as a pharmaceutical composition, a quasi-drug composition, a cosmetic, and a health food. .

  Here, mitochondria and microsomes were prepared as follows.

  After the rats were killed by anesthesia with diethyl ether, the liver was taken out, blood was removed with a homogenizing medium (pH 7.4) and washed several times. The cells were suspended in a homogenizing medium (pH 7.4) of 9 times the amount of liver, and the cells were crushed with a homogenizer. This crushed liquid was centrifuged at 1,000 × g for 10 minutes at 4 ° C. After centrifuging the supernatant at 4 ° C. and 7,000 × g for 10 minutes, microsomes were prepared from the supernatant and mitochondria were prepared from the precipitate.

  The precipitate was suspended in a homogenizing medium (pH 7.4) and centrifuged at 4 ° C. and 7,000 × g for 10 minutes. The obtained precipitate was suspended in 0.4 times HEPES-NaOH of the liver weight and subjected to ultrasonic treatment (BRANSON Model 450 Sonifer) for 5 seconds. This solution was made submitochondoria and used the same day.

  The supernatant was centrifuged at 105,000 × g for 60 minutes at 4 ° C., and the precipitate was suspended in a homogenizing medium (pH 7.4) and then centrifuged under the same conditions. The obtained precipitate was suspended in 100 mM Tris-HCl buffer, 0.7 times the liver weight, as a microsome solution and stored frozen.

  And each test of the following Examples 1-4 was done.

  The 18 types of xanthone derivatives shown in FIGS. 1 to 3 were used for the test. Of these, [Chemical Formula 1] is shown in IYC-2 of FIG. 1, and [Chemical Formula 2] is IYC of FIG. -4, [Chemical Formula 3] is shown in IYC-8 in FIG. 1, [Chemical Formula 4] is shown in IYC-10 in FIG. 2, and [Chemical Formula 5] is shown in FIG. This is shown in IYC-14.

  In Tables 1 to 3 showing the test results, Compd.1 to Compd.18 correspond to IYC-1 to IYC-14 in FIGS.

We measured membrane lipid peroxidation in mitochondria.

Mitochondria were each peroxidized with NADH dependence, NADPH dependence, succinic acid dependence and AsA-Fe ³⁺ , and the lipid peroxidation inhibitory action of each xanthone derivative was measured by the thiorubicuric acid (TBA) method. NADH-dependent and succinic acid-dependent lipid peroxidation is electron transport dependent.

(1) Mitochondrial membrane lipid peroxidation dependent on NADH in the presence of Fe ³⁺ -ADP In a test tube, 10 μl of xanthone solution (DMSO), 500 μl of 100 mM HEPES-NaOH buffer, 100 μl of 20 mM ADP (final concentration 2 mM), 20 mM FeCl ₃ (final concentration 2 mM) was added 100 μl, distilled water 130 μl, 1 mM rotenone (final concentration 10 μM) 10 μl and submitochondoria 50 μl were added and incubated at 37 ° C. for 5 minutes. The reaction was started by adding 100 μl of 1 mM NADH (final concentration 100 μM) and allowed to react at 37 ° C. for 5 minutes. 90 μl of 2% BHT (2,6-Di-t-butyl-4-methylphenol) was added to stop the reaction, and 2 ml of 15% TCA (trichloroaceticacid) -0.375% TBA (2-thiobarbituric acid) -2.5% HCl solution The mixture was heated at 100 ° C. for 15 minutes, centrifuged at 5,000 rpm for 10 minutes, and the absorbance of the supernatant was measured at 535 nm.

  The results of 50% inhibitory concentration (μM) are listed in the item “(1) NADH-dependent” in Table 1. As shown in this table, the xanthone derivative of [Chemical Formula 1] (Compd. 2 in the table) is 2.1, the xanthone derivative of [Chemical Formula 2] (Compd. 4 in the table) is 0.9, and the xanthone of [Chemical Formula 3] The derivative (Compd. 8 in the table) is 27.3, the xanthone derivative (Compd. 10 in the table) is 0.5, and the xanthone derivative (Compd. 14 in the table is 4.1) is 4.1. Yes, it shows very good numerical values for xanthone derivatives having other structures.

(2) Fe ³⁺ -NADPH-dependent mitochondrial membrane lipid peroxidation in the presence of FeDP ^{+ In a} test tube, 10 μl of xanthone, 500 μl of 100 mM HEPES-NaOH buffer, 100 μl of 20 mM ADP (final concentration 2 mM)
Then, 100 μl of 20 mM FeCl ₃ (final concentration 2 mM), 140 μl of distilled water and 50 μl of submitochondoria were added, and the mixture was incubated at 37 ° C. for 5 minutes. 100 μl of 1 mM NADPH (final concentration 100 μM) was added to start the reaction, and the reaction was allowed to proceed at 37 ° C. for 15 minutes. The reaction was stopped as in (1), and the absorbance at 535 nm was measured by the TBA method.

  The result of 50% inhibitory concentration (μM) is described in the item “(2) NADPH-dependent” in Table 1. As shown in this table, the xanthone derivative of [Chemical Formula 1] (Compd. 2 in the table) is 4.8, the xanthone derivative of [Chemical Formula 2] (Compd. 4 in the table) is 1.3, and the xanthone of [Chemical Formula 3] The derivative (Compd. 8 in the table) is 57.8, the xanthone derivative (Compd. 10 in the table) is 1.4, and the xanthone derivative (Compd. 14 in the table) is 2.2. It shows very good numerical values for xanthone derivatives having other structures.

(3) Fe ³⁺ -Mitochondrial membrane lipid peroxidation that depends on succinic acid in the presence of ADP Xanthone 10 μl, 100 mM HEPES-NaOH buffer 500 μl, 20 mM ADP (final concentration 2 mM) 100 μl, 20 mM FeCl ₃ (final) 100 μl of concentration 2 mM), 130 μl of distilled water, 10 μl of 1 mM antimycin (final concentration 10 μM) and 50 μl of submitochondoria were added and incubated at 37 ° C. for 5 minutes. 100 μl of 4 mM sodium succinate (final concentration 400 μM) was added to start the reaction, and the reaction was carried out at 37 ° C. for 60 minutes. The reaction was stopped as in (1), and the absorbance at 535 nm was measured by the TBA method.

  The result of 50% inhibition concentration (μM) is described in the item “(3) Succinate-dependent” in Table 1. As shown in this table, the xanthone derivative of [Chemical Formula 1] (Compd. 2 in the table) is 3.7, the xanthone derivative of [Chemical Formula 2] (Compd. 4 in the table) is 1.3, and the xanthone of [Chemical Formula 3] The derivative (Compd. 8 in the table) is 42.8, the xanthone derivative (Compd. 10 in the table) is 1.0, and the xanthone derivative (Compd. 14 in the table is Compd. 14) is 3.0. It shows very good numerical values for xanthone derivatives having other structures.

(4) Lipid peroxidation of mitochondrial membranes by hydroxy radicals (.OH) generated by low concentrations of ascorbic acid and iron In a test tube, xanthone 10 μl, 100 mM HEPES-NaOH buffer (pH 7.4) 500 μl, 200 mM KCl ( 100 μl of 100 μM FeSO ₄ (final concentration of 10 μM), 140 μl of distilled water and 50 μl of submitochondoria were added to give a reaction solution. After incubating at 37 ° C. for 5 minutes, 100 μl of 2 mM AsA (ascorbic acid) (final concentration 200 μM) was added and reacted at 37 ° C. for 20 minutes. The reaction was stopped as in (1), and the absorbance at 535 nm was measured by the TBA method.

  The result of 50% inhibition concentration (μM) is described in the item “(4) Ascorbate-induced” in Table 1. As shown in this table, the xanthone derivative of [Chemical Formula 1] (Compd. 2 in the table) is 7.1, the xanthone derivative of [Chemical Formula 2] (Compd. 4 in the table) is 5.7, and the xanthone of [Chemical Formula 3] The derivative (Compd. 8 in the table) is 18.3, the xanthone derivative (Compd. 10 in the table) is 5.5, and the xanthone derivative (Compd. 14 in the table) is 5.6. It shows very good numerical values for xanthone derivatives having other structures.

We measured lipid peroxidation in microsomes.

The microsomes were peroxidized with NADPH-dependent, NADH-dependent, AsA-Fe ₃ ⁺ , t-BOOH, CCl ₄ and the lipid peroxidation inhibitory action was measured by the TBA method. NADPH-dependent lipid peroxidation is NADPH-cytochrome-P-450 reductase dependence, NADH dependence is NADH-cytochrome b ₅ reductase dependence, and t-BOOH and CCl ₄ are P-dependent.

(1) Microsomal lipid peroxidation dependent on NADPH in the presence of Fe ³⁺ -ADP In a test tube, 10 μl of xanthone, 500 μl of 100 mM Tris-HCl buffer (pH 7.5), 100 μl of 20 mM ADP (final concentration 2 mM), 1.2 mM FeCl ₃ (final concentration 120 μM) 100 μl, distilled water 140 μl and microsome 50 μl were added to prepare a reaction solution. After incubating at 37 ° C. for 5 minutes, 100 μl of 1 mM NADPH (final concentration 100 μM) was added and reacted at 37 ° C. for 15 minutes. The reaction was stopped as in the case of mitochondria, and the absorbance at 535 nm was measured by the TBA method.

  The result of 50% inhibitory concentration (μM) is described in the item “(5) NADPH-dependent” in Table 2. As shown in this table, the xanthone derivative of [Chemical Formula 1] (Compd. 2 in the table) is 18.9, the xanthone derivative of [Chemical Formula 2] (Compd. 4 in the table) is 6.4, and the xanthone of [Chemical Formula 3] The derivative (Compd. 8 in the table) is 64.4, the xanthone derivative (Compd. 10 in the table) is 0.7, and the xanthone derivative (Compd. 14 in the table) is 1.3. It shows very good numerical values for xanthone derivatives having other structures.

(2) Microsomal lipid peroxidation that depends on NADH in the presence of Fe ³⁺ -ADP In a test tube, 10 μL of xanthone, 500 μl of 100 mM Tris-HCl buffer (pH 7.5), 100 μl of 20 mM ADP (final concentration 2 mM), 1.2 mM FeCl ₃ (final concentration 120 μM) 100 μl, distilled water 140 μl and microsome 50 μl were added to prepare a reaction solution. After incubating at 37 ° C. for 5 minutes, 100 μl of 1 mM NADH (final concentration 100 μM) was added and reacted at 37 ° C. for 60 minutes. Similarly, the reaction was stopped, and absorbance at 535 nm was measured by the TBA method.

  The result was the same as the item (1).

(3) Microsomal lipid peroxidation by hydroxy radicals (.OH) produced by low concentrations of ascorbic acid and iron In test tubes, 10 μl of xanthone, 500 μl of 40 mM phosphate buffer (pH 6.0), 1.2 mM FeCl ₃ (final concentration) 120 μM), 100 μl of 900 mM (final concentration 90 mM) KCl, 140 μl of distilled water, and 50 μl of microsome were added to prepare a reaction solution. After incubation at 37 ° C. for 5 minutes, 100 μl of 5 mM AsA (ascorbic acid) (final concentration 500 μM) was added and reacted at 37 ° C. for 15 minutes. Similarly, the reaction was stopped, and absorbance at 535 nm was measured by the TBA method.

  The result of 50% inhibition concentration (μM) is described in the item “(7) Ascorbate-induced” in Table 2. As shown in this table, the xanthone derivative of [Chemical Formula 1] (Compd. 2 in the table) is 2.4, the xanthone derivative of [Chemical Formula 2] (Compd. 4 in the table) is 1.6, and the xanthone of [Chemical Formula 3] The derivative (Compd. 8 in the table) is 45.7, the xanthone derivative (Compd. 10 in the table) is 0.6, the xanthone derivative (Compd. 14 in the table) is 0.7, It shows very good numerical values for xanthone derivatives having other structures.

(4) p-450-dependent lipid peroxidation by CCl _{4 In a} test tube, 10 μl xanthone, 2 units glucose-6-phosphate-dehydrogenase, 200 mM K phosphate buffer (pH 7.4) containing 0.4 mM EDTA, 500 μl, 100 mM glucose- 100 μl of 6-phosphate, disodium salt (final concentration 10 mM), 240 μl of distilled water and 50 μl of microsome were added to prepare a reaction solution. After incubating at 37 ° C. for 2 minutes, 10 μl of 2M CCl ₄ (carbon tetrachloride) / EtOH (final concentration 20 mM) was added, and further incubated at 37 ° C. for 2 minutes, and then 100 μl of 100 mM (final concentration 1 mM) NADP ⁺ was added and incubated at 37 ° C. for 20 minutes. Reacted for 1 minute. Similarly, the reaction was stopped, and absorbance at 535 nm was measured by the TBA method.

The result of 50% inhibition concentration (μM) is described in the item of “(8) CCl ₄ -induced” in Table 2. As shown in this table, the xanthone derivative of [Chemical Formula 1] (Compd. 2 in the table) is 3.3, the xanthone derivative of [Chemical Formula 2] (Compd. 4 in the table) is 2.9, and the xanthone of [Chemical Formula 3] The derivative (Compd. 8 in the table) is 13.5, the xanthone derivative of [Chem. 4] (Compd. 10 in the table) is 3.5, and the xanthone derivative of [Chem. 5] (Compd. 14 in the table) is 2.3. It shows very good numerical values for xanthone derivatives having other structures.

(5) p-450-dependent lipid peroxidation by t-BOOH To a test tube, 10 μl of xanthone, 500 μl of 200 mM K phosphate buffer (pH 7.4), 430 μl of distilled water, and 50 μl of microsome were added to obtain a reaction solution. After incubating at 37 ° C. for 5 minutes, 100 μl of 100 mM t-BOOH (tert-butylhydroperooxide: final concentration 10 mM) was added and reacted at 37 ° C. for 30 minutes. Similarly, the reaction was stopped, and absorbance at 535 nm was measured by the TBA method.

  The result of 50% inhibition concentration (μM) is described in the item “(9) t-BuOOH-induced” in Table 2. As shown in this table, the xanthone derivative of [Chemical Formula 1] (Compd. 2 in the table) is 58.5, the xanthone derivative of [Chemical Formula 2] (Compd. 4 in the table) is 29.5, and the xanthone of [Chemical Formula 4] The derivative (Compd. 10 in the table) is 35.7, and the xanthone derivative (Compd. 14 in the table is 47.2) is 47.2, which shows very good numerical values for xanthone derivatives having other structures. Yes.

The radical removal effect was measured.

(1) DPPH radical removal action Direct radical removal action was measured using DPPH (diphenyl-p-picrylhydradical) radical, which is a neutral radical.

  Add 20 μl of xanthone, 400 μl of 250 mM acetate buffer, pH 5.5 and 390 μl of ethanol to a test tube, incubate at 37 ° C for 5 minutes, add 200 μl of 250 μM (final concentration 50 μM), react at 30 ° C for 30 minutes, and measure absorbance at 517 nm did. Immediately after the measurement, 30 μm of 3,000 ppm BHT was added, the mixture was kept at 30 ° C. for 30 minutes, and the absorbance at 517 nm was measured.

The result of 50% inhibition concentration (μM) is described in the item “(10) CCl ₄ -induced” in Table 3. As shown in this table, the xanthone derivative of [Chemical Formula 1] (Compd. 2 in the table) is 9.9, the xanthone derivative of [Chemical Formula 2] (Compd. 4 in the table) is 5.8, and the xanthone of [Chemical Formula 4] The derivative (Compd. 10 in the table) is 12.6, and the xanthone derivative (Compd. 14 in the table) is 7.2, showing very excellent numerical values for xanthone derivatives having other structures. Yes.

(2) Superoxide anion (O ₂ ⁻ ) removal action Among various active oxygens, particularly the superoxide anion (O ₂ ⁻ ), the elimination action of O ₂ ⁻ produced by xanthine oxidase (XOD) was measured.

Add 0.5 ml of 100 mM Na phospate buffer, pH7.8 containing 1.1 x 10 ^-3 units XOD, 0.1 mM EDTA, 0.1% BSA (bovine serum albumin fatty acid free), 50 µM nitroblue tetrazolium (NBT) to 10 µl xanthone. It was set as the reaction liquid. After incubating at 37 ° C. for 5 minutes, 0.5 ml of 0.2 mM xanthine (final concentration 0.1 mM) was added as a substrate to start production of superoxide anion. After 20 minutes of reaction, 35 μl of 6 mM CuCl ₂ (final concentration 200 μM) was added to stop the reaction, and the reduced NBT was measured by absorbance at 560 nm.

The result of 50% inhibition concentration (μM) is described in the item “(11) CCl ₄ -induced” in Table 3. As shown in this table, the xanthone derivative of [Chemical Formula 1] (Compd. 2 in the table) is 51.8, the xanthone derivative of [Chemical Formula 2] (Compd. 4 in the table) is 5.7, and the xanthone of [Chemical Formula 4] The derivative (Compd. 10 in the table) is 5.2, and the xanthone derivative of [Chemical Formula 5] (Compd. 14 in the table) is 6.3, which shows very good numerical values for xanthone derivatives having other structures. Yes.

  For the xanthone derivative of Compd. 2 [Chemical Formula 1], mitochondrial peroxidation and enzyme activity were measured.

  In mitochondria, active oxygen is generated by NADH dehydrogenase or coenzyme Q cycle, and as a result, NADH dehydrogenase and complex III are inactivated by oxidative stress. Non-enzymatic generation of hydroxyl radicals by dihydroxyfumarate (DHF) reduces mitochondrial enzyme activity. The enzyme activity protective action of the xanthone derivative was investigated.

To 100 μl of xanthone, 7.7 ml of 50 mM phosphate buffer (pH 7.4), 1 ml of 1 mM FeCl ₃ (final concentration of 100 μM) and 1 ml of 10 mM ADP (final concentration of 1 mM) were added and preincubated at 37 ° C. After 5 minutes, 30 mM DHF (final concentration 300 μM) and 100 μl of mitochondria were added to initiate the reaction. 1 ml was sampled every 30 minutes, and its NADH-cytochrome c reductase activity and succinate-cytochrome c activity were measured.

The enzyme activity was measured continuously for 3 minutes at an absorbance of 550 nm by adding 950 μl of a solution obtained by adding 10 mM NaN ₃ and cytochrome c to 50 mM phosphate buffer and 66 μl of 2 mM NADH or 200 mM succinate to 1 ml of the sampling solution. This was continuously performed for 2 hours from the start of the reaction.

  The results are shown in the graph of the effect of [Chemical Formula 2] on the oxidative damage of the mitochondrial respiratory chain by dihydroxyfumaric acid in FIG. In the figure, the enzyme is not inactivated by the control of (X) DHF-free. When a xanthone derivative of Compd. 2 [Chemical Formula 1] is added to (Y) in the figure, the deactivation is suppressed and the activity is protected. Deoxidizes when peroxidized with DHF. The same protective action was observed for the xanthone derivative of Compd.4 [Chemical 2], the xanthone derivative of Compd.10 [Chemical 4], and the xanthone derivative of Compd.14 [Chemical 5].

  It can prevent various diseases and aging, and can be applied to various uses such as antioxidants, pharmaceutical compositions, quasi-drug compositions, cosmetics, and health foods.

The figure explaining the structure of the xanthone derivative (IYC-1 to IYC-8) used for the test. The figure explaining the structure of the xanthone derivative (IYC-9-IYC-16) used for the test. The figure explaining the structure of the xanthone derivative (IYC-17-IYC-18) used for the test. The graph which shows the effect of [Chemical Formula 2] with respect to the oxidative damage of the mitochondrial respiratory chain by dihydroxy fumarate.

Claims (5)

An antioxidant comprising at least one of xanthone derivatives represented by the following general formulas [Chemical Formula 1], [Chemical Formula 2], [Chemical Formula 3], [Chemical Formula 4], and [Chemical Formula 5].

It contains at least one of the xanthone derivatives represented by the above general formulas [Chemical Formula 1], [Chemical Formula 2], [Chemical Formula 3], [Chemical Formula 4], and [Chemical Formula 5], and has an antioxidant effect. Pharmaceutical composition. It contains at least one of the xanthone derivatives represented by the above general formulas [Chemical Formula 1], [Chemical Formula 2], [Chemical Formula 3], [Chemical Formula 4], and [Chemical Formula 5], and has an antioxidant effect. Quasi-drug composition. It contains at least one of the xanthone derivatives represented by the above general formulas [Chemical Formula 1], [Chemical Formula 2], [Chemical Formula 3], [Chemical Formula 4], and [Chemical Formula 5], and has an antioxidant effect. Cosmetics. It contains at least one of the xanthone derivatives represented by the above general formulas [Chemical Formula 1], [Chemical Formula 2], [Chemical Formula 3], [Chemical Formula 4], and [Chemical Formula 5], and has an antioxidant effect. healthy food.

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