JP2006298807A - Novel compound and use thereof - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide an antioxidant and/or an anticancer agent containing as an active ingredient a novel compound separated from marine life, an ester thereof, a salt thereof, or these. <P>SOLUTION: The antioxidant and/or an anticancer agent contains as an active ingredient a compound represented by formula (1), formula (2), or a like formula, an ester thereof, a salt thereof, or these. <P>COPYRIGHT: (C)2007,JPO&INPIT

Description

The present invention relates to a novel compound isolated from marine organisms, an ester thereof or a salt thereof and use thereof.
Specifically, the present invention relates to an effective prophylactic or therapeutic agent for various diseases and cancers having an effective antioxidant effect, which contains the above-mentioned compound or an ester thereof or a salt thereof.

  In general, many organisms inhabit aerobic conditions, for example, in the air, in the ground where the air circulates, in water in which the air is dissolved. Since this kind of living organisms uses the oxidizing action of oxygen in the atmosphere to obtain energy for survival, oxygen is indispensable for survival. However, when oxygen is used in a living body, a molecular species (active oxygen species) that is highly reactive, that is, highly toxic to the living body, is generated in the living body. Therefore, the living body has a self-protection mechanism that can suppress the generation of reactive oxygen species or can eliminate the generated reactive oxygen species.

  However, the suppression and elimination of the generation of reactive oxygen species in the living body is possible under the delicate balance of the self-protection mechanism that performs these, and when the balance is lost due to various causes, the biological component is changed by the reactive oxygen species. Get oxidative injury. The target of reactive oxygen species in the living body is mainly lipids, nucleic acids, proteins and the like, and when these are damaged, various diseases are caused. That is, reactive oxygen species generated in the living body are various diseases such as cerebral disorders such as schizophrenia and manic depression, adult respiratory distress syndrome, arteriosclerosis, hypertension, circulatory disorders such as hypertension, nephritis, Kidney disorders such as renal failure, liver disorders such as alcoholic hepatitis and cirrhosis, diabetic complications such as cataracts and peripheral nerve disorders, gastrointestinal disorders such as gastric and duodenal ulcers, other rheumatoid arthritis, carcinogenesis, accelerated aging, UV damage Is also known to be involved. It is important for the prevention and treatment of diseases to eliminate reactive oxygen species that are one of the causes of these various diseases. Therefore, there is an expectation for the use of an antioxidant capable of eliminating active oxygen species.

  Conventionally, an antioxidant is generally blended in an appropriate amount in cosmetics and foods in order to prevent deterioration due to oxidation of oil and fat components generally contained in cosmetics and foods. As the antioxidant, a synthetic antioxidant, a natural antioxidant, or a combination thereof can be used. For example, butylhydroxytoluene (BHT) or dibutylhydroxyanisole is used as a synthetic antioxidant, and ascorbic acid, tocopherol, or the like is used as a natural antioxidant. Although conventional synthetic antioxidants such as the above-mentioned BHT have excellent antioxidant ability, natural antioxidants, which have few problems in terms of safety, can be expected to be applied to cosmetics and foods. In vivo efficacy is low. Under such circumstances, it is desired to develop an antioxidant as a pharmaceutical effective and safe for a living body.

  On the other hand, conventionally, chemotherapy using an anticancer agent occupies an important position as a cancer treatment method together with surgical treatment and radiotherapy, and various anticancer agents have been provided. Among anticancer agents, many anticancer agents derived from natural products such as adriamycin, mitomycin, bleomycin, vincristine and the like are known. However, conventional anticancer drugs still have problems in terms of unsatisfactory treatment results and serious side effects, and there are also problems of multidrug resistance, so the appearance of even better anticancer drugs is required. It has been. A compound having an unprecedented novel structure exhibits a new mechanism of action, and is likely to be a more effective drug with fewer side effects. Moreover, the effect as pharmaceuticals other than an anticancer agent can also be anticipated by the action mechanism.

  An object of the present invention is to provide a naturally-occurring novel compound having excellent antioxidant action and anticancer action.

で表される化合物又はそのエステル或いはその塩、
２． １．に記載の化合物又はそのエステル或いはその塩を有効成分として含有する医薬、
３． １．に記載の化合物又はそのエステル或いはその塩を有効成分として含有する抗酸化剤、
４． １．に記載の化合物又はそのエステル或いはその塩を有効成分として含有する抗癌剤、及び
５． １．に記載の化合物又はそのエステル或いはその塩を有効成分として含有する脳障害、循環器障害、腎障害、肝障害、糖尿病合併症、消化管障害、慢性関節リウマチ、癌、
老化促進又は紫外線障害の予防或いは治療剤に関するものである。 As a result of intensive studies to solve the above problems, the present inventors have found that a novel compound isolated from marine organisms has an anticancer activity and an antioxidant activity, and completed the present invention. .
That is, the present invention
1. Formula (1), (2), (3) or (4)

Or a compound thereof or a salt thereof,
2. 1. A pharmaceutical comprising the compound or ester or salt thereof as described in
3. 1. An antioxidant containing as an active ingredient the compound or ester or salt thereof described in 1.
4). 1. 4. an anticancer agent comprising the compound or its ester or its salt as described in 1 above as an active ingredient, and 1. A brain disorder, cardiovascular disorder, renal disorder, hepatic disorder, diabetic complications, gastrointestinal tract disorder, rheumatoid arthritis, cancer,
The present invention relates to a preventive or therapeutic agent for aging promotion or UV damage.

  The novel compound or ester or salt thereof of the present invention has excellent antioxidant action and anticancer action and can be effectively used as a pharmaceutical product or a related product. In addition, the novel compound of the present invention can be easily obtained in a sufficient amount from naturally occurring seaweeds such as the seaweed Togemoku of the family Hawkidae. Since the seaweed Togemoku has not been used so far, it has abundant resources and can be mass-produced if necessary.

で表される化合物又はそのエステル或いはその塩である。
式（１）、（２）、（３）又は（４）で表される化合物は、海洋生物から分離することにより得ることができる。海洋生物の具体例としては、ホンダワラ科に属する海藻トゲモクが挙げられ、分離方法としては、抽出が挙げられる。
抽出方法としては、温水又は有機溶媒での抽出が挙げられ、好ましくは有機溶媒による抽出が挙げられる。
使用する有機溶媒としては、メタノール、エタノール、酢酸エチル、アセトニトリル、クロロホルム及び上述の２種以上の混合溶媒が挙げられる。抽出率を高めるために抽出工程を２回以上繰り返すのが好ましい。また、抽出工程を２回以上繰り返す場合、各抽出工程に使用する溶媒は同一又は異なっていてもよい。抽出に用いる海洋生物は新鮮なもの又は乾燥したものを用いることができる。得られた抽出溶液から溶媒を留去するために、例えば、遠心分離機又は回転式蒸発濃縮機のような分離、濃縮手段を用いることができる。 The compound of the present invention or an ester thereof or a salt thereof is represented by the following formula (1), (2), (3) or (4).

Or a salt thereof or a salt thereof.
The compound represented by the formula (1), (2), (3) or (4) can be obtained by separation from marine organisms. Specific examples of marine organisms include seaweed Togemoku, which belongs to the family Honda, and the separation method includes extraction.
Examples of the extraction method include extraction with warm water or an organic solvent, preferably extraction with an organic solvent.
Examples of the organic solvent to be used include methanol, ethanol, ethyl acetate, acetonitrile, chloroform, and the above-mentioned two or more mixed solvents. In order to increase the extraction rate, the extraction process is preferably repeated twice or more. Moreover, when repeating an extraction process twice or more, the solvent used for each extraction process may be the same or different. The marine organisms used for extraction can be fresh or dried. In order to distill off the solvent from the obtained extracted solution, for example, a separation / concentration means such as a centrifugal separator or a rotary evaporator / concentrator can be used.

  The extract obtained through these steps can be used directly for various applications such as food, cosmetics, and pharmaceuticals, but if necessary, the extract is dissolved in an organic solvent such as ethanol. The substance can be purified by removing insoluble substances or by dissolving in a solvent having a different polarity and fractionating. In the case of further purification, the compound represented by the formula (1), (2), (3) or (4) can be isolated by silica gel column chromatography or the like.

The ester of the compound represented by the formula (1), (2), (3) or (4) can be produced by acylating the hydroxyl group of each compound, but has two or more hydroxyl groups. The number of hydroxyl groups to be acylated in the compound can be arbitrarily selected within the range of the number of hydroxyl groups possessed, and the site of the hydroxyl group to be acylated can also be arbitrarily selected.
Specific ester types include those in which the hydroxyl group of the compound is acetylated, palmitoylated, propanoylated, pivaloylated, succinylated, fumarylated, alanylated, and dimethylaminomethylcarbonylated. When two or more places are esterified by the compound which has one or more, each ester may be the same or different.

The above-mentioned ester can be produced by reacting a compound represented by the formula (1), (2), (3) or (4) under known esterification conditions.
Specifically, it can be produced by reacting a compound represented by the formula (1), (2), (3) or (4) with an acyl halide or acid anhydride in the presence of a base.
Examples of the base include inorganic bases such as potassium carbonate, sodium carbonate, potassium hydrogen carbonate and sodium hydrogen carbonate, and organic bases such as triethylamine, pyridine and collidine.
In addition, the above-mentioned ester can also be used as a prodrug.

Examples of the salt of the compound represented by the formula (1), (2), (3) or (4) or an ester thereof include, for example, metal salts, ammonium salts, salts with organic bases, salts with inorganic acids, and organic acids. And salts with basic or acidic amino acids. Preferable examples of the metal salt include alkali metal salts such as sodium salt and potassium salt; alkaline earth metal salts such as calcium salt, magnesium salt and barium salt; aluminum salt and the like. Preferable examples of the salt with an organic base include, for example, trimethylamine, triethylamine, pyridine, picoline, 2,6-lutidine, ethanolamine, diethanolamine, triethanolamine, cyclohexylamine, dicyclohexylamine, N, N′-dibenzylethylenediamine. And the like. Preferable examples of the salt with inorganic acid include salts with hydrochloric acid, hydrobromic acid, nitric acid, sulfuric acid, phosphoric acid and the like. Preferable examples of the salt with organic acid include formic acid, acetic acid, trifluoroacetic acid, phthalic acid, fumaric acid, oxalic acid, tartaric acid, maleic acid, citric acid, succinic acid, malic acid, methanesulfonic acid, and benzenesulfone. And salts with acid, p-toluenesulfonic acid and the like. Preferable examples of salts with basic amino acids include salts with arginine, lysine, ornithine and the like, and preferable examples of salts with acidic amino acids include salts with aspartic acid and glutamic acid, for example. It is done.

Of these, pharmaceutically acceptable salts are preferred. For example, when the compound has an acidic functional group, an inorganic salt such as an alkali metal salt (eg, sodium salt, potassium salt), an alkaline earth metal salt (eg, calcium salt, magnesium salt, barium salt), When an ammonium salt or the like has a basic functional group in the compound, for example, a salt with an inorganic acid such as hydrochloric acid, hydrobromic acid, nitric acid, sulfuric acid, phosphoric acid, or acetic acid, phthalic acid, fumaric acid, Preference is given to salts with organic acids such as oxalic acid, tartaric acid, maleic acid, citric acid, succinic acid, methanesulfonic acid, p-toluenesulfonic acid.

  The compound of the present invention or an ester thereof or a salt thereof is based on its antioxidant action (for example, antilipid peroxidation action), and in the medical field, for example, brain disorders such as schizophrenia and manic depression, adult respiratory distress syndrome, Cardiovascular disorders such as arteriosclerosis, hypertension, thrombosis, renal disorders such as nephritis and renal failure, liver disorders such as alcoholic hepatitis and cirrhosis, diabetic complications such as cataracts and peripheral nerve disorders, gastric ulcers and duodenal ulcers It is useful as a preventive or therapeutic agent for gastrointestinal disorders, other rheumatoid arthritis, cancer, aging promotion, and UV damage.

  The compound of the present invention or an ester thereof or a salt thereof is used as an anti-ultraviolet ray preventive agent for sunburn prevention, cosmetics (for example, lotion, essence, emulsion, cream, haptic agent, paste agent, gel agent, powder, foundation, lotion, Cosmetics such as packs, skin cleansers such as soaps, facial cleansers, body soaps, hair care products such as shampoos, rinses, hair nicks, hair nourishing agents, bath preparations, whitening agents, UV care products, etc.), cosmetics (eg creams, It can be applied to basic cosmetics such as emulsions, lotions, face wash, packs, makeup cosmetics such as lipsticks, foundations, body soaps, soaps, etc. Also, hair tonics, hair liquids, hair set lotions, Hair products such as hair blowing agents, hair creams and hair coats Npu, rinsing, can also be used as an active ingredient, such as hair care products, etc.) toiletries such for hair such as hair treatment.

  Furthermore, the compound of the present invention or an ester thereof or a salt thereof is used as a highly safe antioxidant, such as various foods and drinks such as retort food, canned food, bottled food, kneaded food, dried food, powdered food, processed food, and smoked food. It can be widely applied to breads, rice, beverages, liquors, confectionery and the like.

  In addition, the compound of the present invention or an ester thereof or a salt thereof is based on its anticancer activity, and cancer (eg, colorectal cancer, breast cancer, lung cancer, prostate cancer, esophageal cancer, stomach cancer, liver cancer, biliary tract cancer, spleen cancer, kidney) Cancer, bladder cancer, uterine cancer, ovarian cancer, testicular cancer, thyroid cancer, pancreatic cancer, colon cancer, brain tumor, blood tumor, etc.) and is useful as a preventive or therapeutic agent, particularly as a preventive or therapeutic agent for colon cancer Useful.

The present invention provides a pharmaceutical composition or veterinary pharmaceutical composition containing an effective amount of a compound represented by the formula (1), (2), (3) or (4) or an ester or salt thereof.
Examples of the administration form of the compound of the present invention or an ester thereof or a salt thereof include parenteral administration by injection (subcutaneous, intravenous, intramuscular, intraperitoneal injection), ointment, suppository, aerosol, etc., tablet, capsule Oral administration by granules, pills, syrups, solutions, emulsions, suspensions and the like can be mentioned.
The above-mentioned pharmaceutical or veterinary pharmaceutical composition containing the compound of the present invention or an ester thereof or a salt thereof is about 0.01 to 99.5% of the compound of the present invention based on the weight of the total composition, preferably about Contains 0.1-30%.

Other pharmaceutically or veterinary active compounds may be included in the compound of the present invention or ester or salt thereof or in addition to the composition containing the compound or ester or salt thereof.
In addition, these compositions may contain a plurality of the compounds of the present invention, their esters or their salts.
The clinical dose of the compound of the present invention or an ester thereof or a salt thereof varies depending on age, body weight, patient sensitivity, symptom level, etc., but an effective dose is usually 0.003 to 1.5 g per day for an adult,
Preferably, it is about 0.01 to 0.6 g. However, if necessary, an amount outside the above range can be used.
The compounds of the invention or their esters or their salts are formulated for administration by conventional pharmaceutical means.

That is, tablets, capsules, granules, pills for oral administration are excipients such as sucrose, lactose, glucose, starch, mannit; binders such as hydroxypropylcellulose, syrup, gum arabic, gelatin, sorbit , Tragacanth, methylcellulose, polyvinylpyrrolidone; disintegrants such as starch, carboxymethylcellulose or calcium salts thereof, microcrystalline cellulose, polyethylene glycol; lubricants such as talc, magnesium or calcium stearate, silica; lubricants such as sodium laurate It is prepared using glycerol or the like.
Injections, solutions, emulsions, suspensions, syrups and aerosols are active ingredient solvents such as water, ethyl alcohol, isopropyl alcohol, propylene glycol, 1,3-butylene glycol, polyethylene glycol; surfactants such as Sorbitan fatty acid ester, polyoxyethylene sorbitan fatty acid ester, polyoxyethylene fatty acid ester, polyoxyethylene ether of hydrogenated castor oil, lecithin; suspending agent, for example, carboxymethyl sodium salt, cellulose derivatives such as methylcellulose, tragacanth, gum arabic, etc. Natural rubbers; prepared using preservatives such as esters of paraoxybenzoic acid, benzalkonium chloride, sorbate and the like.
Examples of ointments that are transdermal preparations include white petrolatum, liquid paraffin, higher alcohol, macrogol ointment, hydrophilic ointment, and aqueous gel base.
Suppositories are prepared using, for example, cacao butter, polyethylene glycol, lanolin, fatty acid triglyceride, coconut oil, polysorbate and the like.

Formulation examples containing the compound of the present invention or an ester thereof or a salt thereof are shown below.
Formulation Example 1
Tablet 10g of the present compound
Lactose 260g
600g microcrystalline cellulose
Corn starch 350g
Hydroxypropylcellulose 100g
CMC-Ca 150g
Magnesium stearate 30g
Total amount 1,500g
After the above ingredients are mixed by a conventional method, 10,000 sugar-coated tablets containing 1 mg of the active ingredient are produced.
Formulation Example 2
Capsule 10g of the present compound
440 g of lactose
1,000g microcrystalline cellulose
Magnesium stearate 50g
Total amount 1,500g
The above ingredients are mixed by a conventional method and then filled into gelatin capsules to produce 10,000 capsules containing 1 mg of active ingredient in one capsule.
Formulation Example 3
Soft capsule 10g of the present compound
PEG400 479g
Saturated fatty acid triglyceride 1,500 g
Peppermint oil 1g
Polysorbate 80 10g
Total amount 2,000g
After the above ingredients are mixed, a No. 3 soft gelatin capsule is filled by a conventional method to produce 10,000 capsules containing 1 mg of the active ingredient in one capsule.
Formulation Example 4
Ointment Compound of the present invention 1.0 g
Liquid paraffin 10.0g
Cetanol 20.0g
White petrolatum 68.4g
Ethylparaben 0.1g
l-Menthol 0.5g
Total amount 100.0g
The above ingredients are mixed by a conventional method to make a 1% ointment.
Formulation Example 5
Suppository compound of the present invention 1 g
Whittepsol H15 * 478g
Whittepsol W35 * 520g
Polysorbate 80 1g
Total amount 1,000g
“Trademark name of triglyceride compound Witepsol”
The above ingredients are melt-mixed in a conventional manner, poured into a suppository container and solidified by cooling to produce 1,000 1 g suppositories containing 1 mg of the active ingredient.
Formulation Example 6
Injection preparation Compound of the present invention 1 mg
5mL distilled water for injection
When used, dissolve.

Hereinafter, although an example is given and the present invention is explained in full detail, the present invention is not limited to these examples at all.
Example 1: Extraction and purification of compounds (1) and (2) 2.3 kg of the collected seaweed seaweed Togemoku (scientific name: Sargassum micracanthum) was extracted with methanol (3 L × 3 times), and the solvent was distilled off. A methanol extract (442 g) was obtained. Chloroform-methanol mixture (3: 1) was added to the extract to precipitate and remove impurities such as salts, and the solvent was evaporated to concentrate. This oily substance was separated by silica gel column chromatography (developing solvent: hexane / ethyl acetate (4/1, 2/1, 1/1), methanol) to obtain two fractions (A1, A2). A1 (39.6 g) was separated by silica gel column chromatography (developing solvent: hexane / ethyl acetate (4/1, 2/1, 1/1), ethyl acetate, methanol) and separated into four fractions (A1-1 to A1-1). A1-4) was obtained. A1-2 (2.67 g) was separated by reverse phase silica gel column chromatography (developing solvent: methanol / water (5/1, 8/1), methanol / acetone (3/1)), and three fractions (A1 -2-1, A1-2-2, A1-2-3). For fraction A1-2-1, silica gel column chromatography (developing solvent: hexane / ethyl acetate (4/1, 3/1, 1 /
1), ethyl acetate), and 4 fractions (A1-2-1-1 to A1-2-1-4) were obtained. Fraction A1-2-1-3 was purified by medium pressure liquid chromatography (developing solvent: hexane / ethyl acetate (3/1)) to obtain 8.0 mg of compound (1) and 10.7 mg of compound (2). It was.

Example 2: Chemical structure analysis of compounds (1) and (2) Properties of compound (1) obtained in Example 1 are shown below. Compound (1) was found to have a molecular formula of C ₂₇ H ₃₈ O ₄ from high resolution electron impact mass spectrometry (HREIMS). Also,
In the measurement of the infrared absorption spectrum, (film) V _max 1645, 171
0. ¹ H-NMR spectrum (CDCl ₃ ) and ¹³ C-NMR spectrum (CDCl ₃ ) are shown in Table 1. From these results, it was confirmed that the structure of the compound (1) was a compound represented by the formula (1).
The properties of the compound (2) obtained in Example 1 are shown below. Compound (2) was found to have a molecular formula of C ₂₇ H ₄₀ O ₄ from high resolution electron impact mass spectrometry (HREIMS).
In the measurement of infrared absorption spectrum, (film) V _max 1710 (
Carbonyl group). ^{The 1} H-NMR spectrum (CDCl ₃ ) and ¹³ C-NMR spectrum (CDCl ₃ ) are shown in Table 2. From these results, the structure of the compound (2) has the formula (
It confirmed that it was a compound represented by 2).

Example 3: Extraction and purification of compounds (3) and (4) Silica gel column chromatography (developing solvent: hexane / acetone (8/1, 6) for fraction A-1-2-3 obtained in Example 1 / 1, 4/1), acetone), reverse phase silica gel column chromatography (developing solvent: methanol / water (19/1), acetone), silica gel column chromatography (developing solvent: hexane / acetone (5/1, 3). / 1), acetone), and silica gel column chromatography (developing solvent: hexane / acetone (4/1, 5/2), acetone), followed by sequential purification to obtain compounds (3) and (4).

Example 4: Chemical structure analysis of compounds (3) and (4) Properties of compound (3) obtained in Example 3 are shown below. Compound (3) was found to have a molecular formula of C ₄₅ H ₇₄ O ₅ from high resolution electron impact mass spectrometry (HREIMS). Also,
For the measurement of infrared absorption spectra, (film) V _max 3460, 174
0 and 1655. ^{The 1} H-NMR spectrum (CDCl ₃ ) and ¹³ C-NMR spectrum (CDCl ₃ ) are shown in Table 3. From these results, the structure of the compound (3) is represented by the formula (3
).
Moreover, the property of the compound (4) obtained in Example 3 is shown below. Compound (4) was found to have a molecular formula of C ₄₅ H ₇₆ O ₅ from high resolution electron impact mass spectrometry (HREIMS).
In the measurement of the infrared absorption spectrum, (film) V _max 3460,
1739, 1655. ^{The 1} H-NMR spectrum (CDCl ₃ ) and ¹³ C-NMR spectrum (CDCl ₃ ) are shown in Table 4. From these results, the structure of the compound (4) is
It confirmed that it was a compound represented by Formula (4).

Example 5: Measurement of antioxidant activity 1
DPPH radical scavenging activity was measured using the compound (2) obtained in Example 1 and the compounds (3) and (4) obtained in Example 2.
(1) Test method Antioxidant activity was measured by measuring the radical scavenging activity of 1,1-diphenyl-2-picrylhydrazyl (DPPH) having a free radical in the molecule by the broth method. That is, 0.5 mL of an ethanol solution of each test substance (100 μg / mL) was added to 1.5 mL of a 0.1 mM DPPH ethanol solution and reacted at 37 ° C. for 20 minutes, and then the absorbance was measured at 517 nm. The rate of decrease in absorbance with respect to the control group (test substance concentration 0 μg / mL) was defined as DPPH radical scavenging activity.
(2) Test results As shown in Table 5 below, the DPPH radical scavenging activity of the compounds (2) to (4) has IC ₅₀ values of 3.0 ± 0.5 and 32.3 ± 12.3, respectively. And 52.6 ± 1.7 μg / mL. From the above, it was found that the compounds (2) to (4) have very strong antioxidant activity.

Example 6: Measurement of antioxidant activity 2
Using the compound (2) obtained in Example 1 and the compounds (3) and (4) obtained in Example 2, the lipid peroxidation inhibitory action was measured.
(1) Measurement method 10 μL of rat liver microsome fraction (protein concentration 30 mg / mL) and 10 μL of ethanol solution of the test substance were added to 920 μL of 50 mM Tris-HCl buffer (pH 7.5) containing 7 mM MgCl ₂ , and 37 ° C. For 5 minutes. Next, a NADPH regeneration system (20 μL of 12 mM ferrous sulfate aqueous solution containing 0.2 M ADP, 10 mM β-NADPH 40 μL containing 12.5 U glucose 6-phosphate dehydrogenase and 160 mM glucose 6-phosphate) was added at 37 ° C. For 10 minutes. After the reaction, 2 mL of 0.25M hydrochloric acid (containing 0.375% thiobarbituric acid and 15% trichloroacetic acid) was added to this solution and reacted in boiling water for 15 minutes. By this reaction, malon produced from lipid peroxide was produced. The amount of dialdehyde was determined by measuring the absorbance at a wavelength of 530 nm. The concentration (IC ₅₀ ) at which lipid peroxidation was inhibited by 50% was determined using the absorbance decreased with respect to the control group (test substance concentration 0 μg / mL) as an index for inhibiting lipid peroxidation.
(2) Test results The lipid peroxidation-inhibiting activities of the compounds (2) to (4) have IC ₅₀ values of 0.95 ± 0.25 and 1.15 ± 0.00, respectively, as shown in Table 6 below. 55 and 44.3 ± 22.2 μg / mL. From the above, it was found that the compounds (2) to (4) have very strong antioxidant activity.

Example 7: Cytotoxicity against cancer cell line Using the compound (2) obtained in Example 1 and the compounds (3) and (4) obtained in Example 2, cytotoxicity against the cancer cell line was measured.
(1) Test method Mouse colon cancer cell line used for the test: colon26-L5 is 10% inactivated fetal bovine serum: FBS (Gibco), 100 U / mL penicillin (Wako), 0.1 mg / mL Of Streptomycin (manufactured by Wako) and 55 μM 2-mercaptoethanol (manufactured by Wako) and passaged and maintained in RPMI-1640 medium (manufactured by Gibco).
Cell proliferation was evaluated using WST-1 Cell Counting Kit (manufactured by Wako). Cancer cells were adjusted to a cell density of 2 × 10 ⁵ cells / mL with RPMI-1640 medium containing 5% FBS, and 0.5 to 2 × 10 ⁴ cells were seeded per well of a 96-well plate (manufactured by Iwaki). . After the cells adhered to the plate, test substances were added at various concentrations, and cultured in an incubator (37 ° C., 5% CO ₂ ) for 24-72 hours. For control, dimethyl sulfoxide (manufactured by Wako) was added in the same manner. After completion of the culture, the medium in each well was replaced with 10% FBS-RPMI-1640 medium containing 10% WST-1, and further cultured for 2 to 2.5 hours. Then, the absorbance at a wavelength of 450 nm was measured with a microplate reader (Towa Lab. Product), the decrease rate of the absorbance with respect to the control was determined and expressed as the cell growth inhibition rate.
(2) Test Results It was found that the compounds (2) to (4) have a very strong cytotoxic activity against mouse colon cancer cells as shown in FIG.

The cell growth-inhibition rate with respect to a mouse | mouth colon cancer cell of a compound (2)-(4) and a cisplatin is shown.

Claims (5)

Formula (1), (2), (3) or (4)

Or a salt or a salt thereof. The pharmaceutical which contains the compound of Claim 1, its ester, or its salt as an active ingredient. The antioxidant containing the compound of Claim 1, its ester, or its salt as an active ingredient. The anticancer agent which contains the compound of Claim 1, its ester, or its salt as an active ingredient. A brain disorder, cardiovascular disorder, renal disorder, liver disorder, diabetic complications, gastrointestinal tract disorder, rheumatoid arthritis, cancer, accelerated aging or ultraviolet rays comprising the compound according to claim 1 or an ester thereof or a salt thereof as an active ingredient Preventive or therapeutic agent for disorders.

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