JP2009001569A - Apoptosis-controlling agent - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a safe apoptosis-controlling agent hardly causing side effects while exhibiting remarkable apoptosis-controlling action. <P>SOLUTION: The apoptosis-controlling agent comprises as an effective ingredient 3-[4-hydroxy-3,5-bis(3-methyl-2-butenyl)phenyl]-2-propenoic acid represented by the formula and/or its physiologically acceptable salt(s). <P>COPYRIGHT: (C)2009,JPO&INPIT

Description

  The present invention relates to apoptosis regulators, in particular, 3- [4-hydroxy-3,5-bis (3-methyl-2-butenyl) phenyl] -2-propenoic acid or a physiologically acceptable salt thereof as an active ingredient. The present invention relates to an apoptosis-regulating agent comprising.

  Recently, in the field of biology, natural death of cells, that is, “apoptosis” has attracted attention. Due to natural providence, in normal individuals, all cells are programmed to follow a series of processes of development, differentiation and maturation, and then die naturally and old cells are sequentially replaced by young cells. Yes. However, in malignant tumors, apoptosis of abnormal cells is promoted in the negative direction, and the generation is repeated indefinitely until the individual dies without the cells being differentiated and matured according to a predetermined program. On the other hand, in acquired immune deficiency syndrome (AIDS) and Alzheimer's disease, normal apoptosis of lymphocytes and neurons is promoted in a positive direction, and the patient exhibits an immunodeficiency or dementia state. In other words, the diseases such as “refractory diseases” and “adult diseases” are caused by apoptosis of normal cells and abnormal cells being promoted in a direction opposite to the original direction for some reason. I can say that. At present, there is a demand for substances that can regulate cell apoptosis, and many researchers have eagerly searched, but no effective substances have yet been discovered. The feature of apoptosis is that, when cells are killed, microscopically, cell atrophy and fragmentation occur, and genetically, DNA fragmentation is observed. These features make apoptosis distinct from other cell deaths.

  In view of such circumstances, an object of the present invention is to provide an apoptosis regulator that does not substantially affect normal apoptosis of normal cells while promoting apoptosis of abnormal cells.

  In order to solve this problem, the present inventors diligently searched for various natural extracts, especially propolis extract. As a result, 3- [4-hydroxy-3,5-bis (3 It was found that -methyl-2-butenyl) phenyl] -2-propenoic acid and its salts have a remarkable apoptosis-regulating action. Propolis extract has long been said to have various physiological actions including antibacterial action, anti-inflammatory action, and antitumor action. Recently, females of modern science have entered propolis extracts. For example, Shinobu Matsuda “Food and Food Ingredients of Japan”, No. 160, pp. 64-73 (1994) ) Describes various physiologically active substances contained in the propolis extract.

  3- [4-Hydroxy-3,5-bis (3-methyl-2-butenyl) phenyl] -2-propenoic acid, which has been confirmed by the present inventors in the presence of propolis extract and apoptosis-controlling effect, is known per se. For example, the same patent applicant disclosed in JP-A-6-256177 that the present compound can be isolated from a propolis extract. However, to the best of the inventors' knowledge, there are no publications that teach or suggest the apoptosis-regulating action of this compound, and 3- [4-hydroxy-3,5-bis (3-methyl-2) as an active ingredient An apoptosis regulator comprising -butenyl) phenyl] -2-propenoic acid or a physiologically acceptable salt thereof is an object of this invention.

  3- [4-Hydroxy-3,5-bis (3-methyl-2-butenyl) phenyl] -2-propenoic acid and physiologically acceptable salts thereof promote apoptosis of abnormal cells in humans It exerts an effect that does not substantially affect normal apoptosis of normal cells. Moreover, according to the results of an acute toxicity test using experimental animals, all of these compounds are extremely low in toxicity.

  Hereinafter, embodiments of the present invention will be described, and 3- [4-hydroxy-3,5-bis (3-methyl-2-butenyl) phenyl] -2-propenoic acid used in the apoptosis regulator of the present invention and The salt has a structure shown in Chemical Formula 1 below. In Chemical Formula 1, X represents a physiologically acceptable cation. Examples of such a cation include a hydrogen ion, a sodium ion, a potassium ion, a calcium ion, a magnesium ion, and an ammonium ion.

  3- [4-Hydroxy-3,5-bis (3-methyl-2-butenyl) phenyl] -2-propenoic acid and its salt used in the present invention may be collected from natural sources or chemically synthesized. As long as it has the desired effect of regulating apoptosis, the origin and origin are not limited. Examples of natural sources include propolis and leaf stems of asteraceae plants such as Kawaramugi, for example, Japanese Patent Laid-Open No. 6-256177, See Zero et al. “Phytochemistry”, Vol. 25, No. 12. 2, 841 to 2,855 (1986) and I Okuno et al., “Chemical Pharmaceutical Bulletin”, Vol. 36, No. 2, 769 to 775 (1988). A method for isolating 3- [4-hydroxy-3,5-bis (3-methyl-2-butenyl) phenyl] -2-propenoic acid is described in detail. For chemical synthesis, for example, the method disclosed in JP-A-60-163841 can be advantageously applied.

  The outline of the method for isolation from natural sources is as follows. First, the raw propolis and the leaf stems of Asteraceae are crushed, and water or organic solvents such as methanol, ethanol, acetone, diethyl ether, ethyl acetate, or a mixture thereof. The resulting extract is concentrated and purified. Conventional methods for concentrating and purifying similar compounds may be applied to the concentration and purification of the extract. Examples of such methods include salting out, dialysis, filtration, concentration, fractional precipitation, crystallization, Gel filtration chromatography, ion exchange chromatography, gas chromatography, high performance liquid chromatography and the like can be mentioned, and these are applied in appropriate combination as required. In order to obtain a physiologically acceptable salt of 3- [4-hydroxy-3,5-bis (3-methyl-2-butenyl) phenyl] -2-propenoic acid, 3- [4-hydroxy- 3,5-bis (3-methyl-2-butenyl) phenyl] -2-propenoic acid, for example, hydroxide such as sodium hydroxide, potassium hydroxide, calcium hydroxide, magnesium hydroxide, ammonium hydroxide It only has to act. The salt thus obtained is better in water solubility than 3- [4-hydroxy-3,5-bis (3-methyl-2-butenyl) phenyl] -2-propenoic acid and processed into a desired form. Excellent handling when used.

  The apoptosis-regulating agent of the present invention contains 3- [4-hydroxy-3,5-bis (3-methyl-2-butenyl) phenyl] -2-propenoic acid and / or its physiologically acceptable as an active ingredient. Salt. These compounds suppress the abnormal apoptosis of normal cells while promoting the apoptosis of abnormal cells in humans, and do not substantially affect normal apoptosis of normal cells. Therefore, when the apoptosis regulator of this invention is ingested or administered to humans, it exerts a remarkable effect on treatment / prevention of various diseases caused by abnormal apoptosis of cells, health recovery, and health maintenance. In the present invention, “regulation” of apoptosis refers to an action that promotes the apoptosis of abnormal cells and suppresses the abnormal apoptosis of normal cells. The action which returns to this state or promotes in a preferable direction.

  The apoptosis regulator of the present invention is not only in the form of 3- [4-hydroxy-3,5-bis (3-methyl-2-butenyl) phenyl] -2-propenoic acid or a salt thereof, but also in liquid, pasty and solid forms. In the form of a composition such as a food product, a cosmetic product and a pharmaceutical product. That is, in the case of food, for example, water, alcohol, starch, protein, fiber, saccharide, lipid, vitamin, mineral, flavoring, coloring, sweetener, seasoning, stabilizer, preservative, etc. In the case of cosmetics, as a composition with raw materials or raw materials that are usually blended in simple foods, the base agent, base, surfactant, foaming agent, wetting agent, thickener, clearing agent, flavoring agent, coloring agent In the case of pharmaceuticals, for example, carriers, excipients, diluents, stabilizers, and, if necessary, other physiological activities The form as a composition with 1 type, or 2 or more types of a substance is also included. Regardless of the above form, the apoptosis regulator of this invention is usually 3- [4-hydroxy-3,5-bis (3-methyl-2-butenyl) phenyl] -2-propenoic acid. Or the salt is contained 0.01weight% or more. In addition, since 3- [4-hydroxy-3,5-bis (3-methyl-2-butenyl) phenyl] -2-propenoic acid and its salt have a property of being easily oxidized, they are usually used in foods, cosmetics or pharmaceuticals. The combined use with the antioxidant to be blended is desirable. Examples of the individual antioxidants include sodium nitrite, L-ascorbic acid, sodium L-ascorbate, sodium hydrogen sulfite, sodium sulfite, isoquercetin, sodium edetate, ersorbic acid, cysteine hydrochloride, γ-oryzanol, catechin Licorice extract, citric acid, sodium citrate, quercetin, soy lecithin, sodium thioglycolate, sodium thiomalate, tocotrienol, tocopherol, vitamin E, sodium pyrosulfite, potassium pyrosulfite, butylhydroxyanisole, butylhydroxytoluene, 1 , 3-butylene glycol, benzotriazole, gallic acid, propyl gallate, sodium metabisulfite, malic acid, rutin, enzyme-treated rutin and the like, Content of about 0.01 to 10 wt% in apoptosis modulating agents of the invention, preferably, to be about 0.1 to 1 wt%, are used alone or in combination as appropriate.

  The method of using the apoptosis regulator of the present invention will be described. The apoptosis regulator of the present invention exhibits a remarkable apoptosis regulating action regardless of whether it is used orally or parenterally. Depending on the purpose of use, for example, for the purpose of disease prevention, it is usually taken or administered orally or transdermally in the form of food or cosmetics, while the purpose is to treat the disease In some cases, it is usually administered orally or parenterally by injection, external preparation or the like in the form of a pharmaceutical product. The dose is usually taken or administered orally so that the dose of the active ingredient is about 1 μg to 10 mg per adult, preferably about 10 μg to 1 mg 1 to 4 times / day or 1 to 5 times / week. Or administered parenterally intradermally, subcutaneously, intramuscularly or transdermally.

  The apoptosis regulator of this invention exhibits a remarkable effect in the treatment and prevention of various diseases caused by abnormal apoptosis of cells. Examples of individual diseases include infectious diseases, immune diseases, and malignant tumors, and these diseases can be effectively treated and prevented by ingesting or administering the apoptosis regulator of the present invention. In addition, the apoptosis regulator of the present invention exerts remarkable effects on health recovery and health maintenance when used by sick and healthy people.

  Next, the effectiveness and toxicity of the apoptosis regulator of this invention will be described with reference to experimental examples.

<Experiment 1>
<Preparation of 3- [4-hydroxy-3,5-bis (3-methyl-2-butenyl) phenyl] -2-propenoic acid and its salt>

<Experiment 1-1>
<Preparation of 3- [4-hydroxy-3,5-bis (3-methyl-2-butenyl) phenyl] -2-propenoic acid>
According to the method described in JP-A-6-256177, 3- [4-hydroxy-3,5-bis (3-methyl-2-butenyl) phenyl] -2-propenoic acid was prepared from Brazilian propolis. .

  That is, 153.3 parts by weight of Brazilian propolis mass was crushed and extracted by adding ethyl acetate, methanol was added, and the resulting precipitate was removed by centrifugation, while the supernatant was concentrated by an evaporator, and ethanol was added. Was added. The newly formed precipitate was removed by centrifugation, and the remaining supernatant was concentrated in the same manner as above, and methanol was added to obtain 63.9 parts by weight of the extract.

  35 parts by weight of this extract was converted to a solid substance, loaded onto a column of “silica gel 60 G650” manufactured by Katayama Chemical Industry, and mixed with hexane and ethyl acetate under a concentration gradient. Then, a fraction eluted with a concentration ratio of hexane and ethyl acetate in the range of 59:41 to 57:43 was collected, loaded onto a column of “Sephadex LH-20” manufactured by Pharmacia, and methanol was added to SV0.16. The fraction eluted at around 1.1 times the gel volume was collected and concentrated to precipitate crystals. The crystals were washed with hexane and dried to obtain 0.28 parts by weight of a crystalline solid.

  A part of this crystalline solid is taken and analyzed by elemental analysis, mass spectrometry, ultraviolet spectroscopy, infrared spectroscopy and nuclear magnetic resonance spectroscopy according to conventional methods. 3 [4-Hydroxy-3,5-bis (3-methyl-2-butenyl) reported in Pharmaceutical Bulletin, Vol. 36, No. 2, 769 to 775 (1988) When compared with the analysis result of phenyl] -2-propenoic acid, all showed good agreement. Thus, the crystalline compound obtained from propolis by the above operation (hereinafter referred to as “compound 1”) is 3- [4-hydroxy-3,5-bis (3-methyl-2-butenyl) phenyl. ] -2-propenoic acid.

<Experiment 1-2>
<Preparation of 3- [4-hydroxy-3,5-bis (3-methyl-2-butenyl) phenyl] -2-propenoate>
Compound 1 prepared by the method of Experiment 1-1 was dissolved in ethanol so as to be about 10% (w / w), and equimolar sodium hydroxide, calcium hydroxide, magnesium hydroxide, or potassium hydroxide was added. Then, it concentrated and dried with the centrifugal concentrator and obtained the solid substance. Compound 1 prepared by the method of Experiment 1-1 and its sodium salt (hereinafter referred to as “Compound 2”), calcium salt (hereinafter referred to as “Compound 3”), magnesium salt (hereinafter referred to as “Compound 3”) prepared in this example. , “Compound 4”) and potassium salt (hereinafter referred to as “compound 5”) were appropriately adjusted to an aqueous solution, subjected to sterilization filtration by a conventional method, and then subjected to the next experiment.

<Experiment 2>
<Apoptosis regulating action>

<Experiment 2-1>
<Apoptotic regulation in abnormal and normal cells>
With respect to the compounds 1 to 5 prepared by the method of Experiment 1, an experiment was carried out to confirm the apoptosis-regulating action in an ultramorphic manner using a system using an adherent malignant tumor cell line and a normal cell line as human abnormal cells.

That is, MEM medium (pH 7.0) supplemented with 10% (v / v) fetal bovine serum was placed in a petri dish, and one of the six types of cell lines shown in Table 1 had a cell density of about 1 × 10 ⁵ cells / ml. And cultured at 37 ° C. for 24 hours in a 5% CO ₂ incubator. The culture is aspirated to remove the supernatant, the cells are washed with phosphate buffered saline (pH 7.2), and then any one of compounds 1 to 5 is added at 0 μg / ml, 25 μg / ml, 50 μg / ml or Fresh MEM medium (pH 7.0) containing 100 μg / ml and supplemented with 10% (v / v) fetal bovine serum was added, and further cultured as described above.

  After 18 hours, the culture was treated with trypsin to detach the cells from the petri dish, washed with physiological saline, centrifuged, and first, according to a conventional method, first, 2.5% (w / v) aqueous glutaraldehyde solution Then, after treatment with 1% (w / v) osmium tetroxide aqueous solution and fixation, an embedded specimen was obtained. Thereafter, the specimen was observed under a transmission electron microscope and photographed. Among the photographed photographs, photographs of gastric cancer cell line HGC and normal skin cell line SF-TY when compound 2 is cultured in a culture medium containing 100 μg / ml are shown in FIGS. 1 and 2, respectively.

  At the same time, the occurrence of apoptosis is observed under an electron microscope. When the incidence of apoptosis is 10% or more of the test cells, “+”, when 1% or more and less than 10%, “±”, The case of less than 1% was determined as “−”. The results are shown in Table 1. In addition, in the effect | action with respect to the cell line used for experiment, since the substantial difference was not recognized between the compounds 1-5, in Table 1, the kind of compound is not distinguished.

  The results in Table 1 and FIG. 1 indicate that compounds 1 to 5 act on tumor cells, which are human abnormal cells, and significantly promote apoptosis thereof. Even when observed under an electron microscope, abnormal cells cultured in the presence of compounds 1 to 5 have, for example, the ultrathin form shown in FIG. (The part that looks black) is fragmented and dispersed within the cell. Such cell atrophy and nuclear fragmentation are phenomena characteristic of apoptosis, and cell atrophy and fragmentation characteristic of apoptosis were observed, which were observed in gastric cancer cell line HGC, colon cancer cell line RPMI-4788. (FERM BP-2429), lung cancer cell line HLC, and hepatocellular carcinoma line PLC / PRF / 5 were remarkable. On the other hand, as can be seen from the results of Table 1 and FIG. 2, in the case of normal cells, such findings were not observed even when the same treatment was performed. These facts indicate that 3- [4-hydroxy-3,5-bis (3-methyl-2-butenyl) phenyl] -2-propenoic acid and its physiologically acceptable salts are It shows that it promotes apoptosis but does not substantially affect apoptosis of normal cells.

<Experiment 2-2>
<Apoptotic regulation in abnormal cells>
With respect to compounds 1 to 5 prepared by the method of Experiment 1, apoptosis-regulating action is confirmed at the DNA level by a system using leukemia cell lines U-937 (ATCC CRL 1593) and HL-60 (ATCC CCL240) as human abnormal cells. The experiment was conducted.

That is, RPMI-1640 medium (pH 7.0) supplemented with 10% (v / v) fetal bovine serum was placed in a culture bottle, and one of the two leukemia cell lines shown in Table 2 was added at a cell density of about 4 × 10 4. ^After seeding at 5 / ml and adding any one of compounds 1 to 5 prepared by the method of Experiment 1 to 0 μg / ml, 25 μg / ml, 50 μg / ml or 100 μg / ml, 5 The cells were cultured at 37 ° C. for 7 hours or 24 hours in a% CO ₂ incubator.

  Cells are collected from the culture by centrifugation, washed with phosphate buffered saline, and a portion thereof is treated in the same manner as in Experiment 2-1, to make an embedded specimen, which is observed under an electron microscope and photographed. I took a picture. Further, a part of the remaining cells was taken, and DNA was extracted with a DNA extraction kit “Easy-DNA Kit” manufactured by Invitrogen. / V) After electrophoresis on an agarose gel, the gel was transferred onto a UV transilluminator and photographed.

  Then, the photographed gel electrophoresis image was examined, and DNA fragmentation characteristic of apoptosis, that is, “+” was displayed when “ladder” was observed, and “−” was displayed when it was not observed. Are summarized in Table 2. In addition, in the effect | action with respect to the leukemia cell strain used for experiment, since the substantial difference was not recognized between the compounds 1-5, in Table 2, the kind of compound is not distinguished. 3 and 4 show photographs of ultra-morphology and DNA gel electrophoresis images extracted from the U-937 cells cultured in a culture medium containing 100 μg / ml of Compound 2 for 24 hours, respectively. Show.

  As shown in FIG. 3, as in the case of the adherent tumor cell used in Experiment 2-1, the suspension tumor cell of this example can be seen, for example, in a cell slightly to the right of the center when Compound 2 is allowed to act. As can be seen, the cells are somewhat more atrophic and their nuclei are becoming fragmented. Moreover, as can be seen in the gel electrophoresis image of FIG. 4, a remarkable ladder was observed in the DNA extracted from the cells. Furthermore, as can be seen in Table 2, the occurrence of ladder was observed for compounds 1 to 5. Dependence on dose and incubation time indicates that the atrophy and fragmentation in the ultrastructure of the cells is undoubtedly due to apoptosis.

<Experiment 3>
<Acute toxicity test>
A group of 10 ddy mice weighing 10 to 20 g, and containing appropriate concentrations of compounds 1 to 5 and 5% (w / w) gum arabic obtained by the methods of Experiments 1-1 and 1-2 in the abdominal cavity Saline was injected or administered orally with a gastric sonde, followed by observation for 7 days to examine the number of deaths, and the LD50 value was determined by the van der Vaerden area method. As a result, LD50 of compounds 1 to 5 was remarkably high at 200 mg / kg or higher by any administration route. This is because 3- [4-hydroxy-3,5-bis (3-methyl-2-butenyl) phenyl] -2-propenoic acid and its salts are usually effective in adults at a dose of several μg / dose. Considering some things, it can be said that the apoptosis regulator of the present invention is very safe.

  Hereinafter, embodiments of the present invention will be described specifically by way of examples.

<healthy food>
150 parts by weight of reduced maltose starch jelly “Mabit” sold by Hayashibara Shoji Co., Ltd. was concentrated under reduced pressure to a moisture content of 15% (w / w), and 13 parts by weight of gelatin was dissolved in 18 parts by weight of water. After mixing 0.05 parts by weight of any one of compounds 1 to 5 obtained by the method of Experiment 1, 1 part by weight of citric acid, 1 part by weight of L-ascorbic acid and appropriate amounts of colorant and flavor, Molded and packaged to obtain 5 types of gummy candy containing compounds 1 to 5, respectively.

  Since it contains an antioxidant, the apoptosis-controlling action of compounds 1 to 5 is unlikely to decrease, and this product having a good texture and flavor is useful as a health food.

<healthy food>
3 parts by weight of gum base is heated and melted until soft, 4 parts by weight of sucrose and 3 parts by weight of maltose powder are added thereto, and 0.001 part by weight of any one of compounds 1 to 5 obtained by the method of Experiment 1 is added to vitamin E. After mixing 0.1 part by weight and an appropriate amount of colorant, they were kneaded, molded, and packaged by a conventional method to obtain 5 types of chewing gums each containing compounds 1 to 5.

  Since it contains an antioxidant, the apoptosis-controlling action of compounds 1 to 5 is unlikely to decrease, and this product having a good texture and flavor is useful as a health food.

<Health drink>
10 parts by weight of skim milk was sterilized by heating at 80 ° C. for 20 minutes, cooled to 40 ° C., 0.3 parts by weight of a starter was added thereto, and fermented at about 37 ° C. for 10 hours. After homogenizing the fermented product, 5 parts by weight of isomaltoligosaccharide syrup, 1 part by weight of sucrose and 2 parts by weight of isomerized sugar syrup were added, and heated to 70 ° C. to sterilize. After cooling, 0.01 parts by weight of each of compounds 1 to 5 obtained by the method of Experiment 1 and malic acid were added and bottled to obtain five types of lactic acid beverages containing compounds 1 to 5, respectively.

  Since it contains an antioxidant, the apoptosis-controlling action of compounds 1 to 5 is unlikely to decrease, and this product with good flavor and taste is useful as a health drink.

<Cosmetics>
0.5 parts by weight of polyoxyethylene behenyl ether, 1 part by weight of polyoxyethylene sorbitol tetraoleate, 1 part by weight of glyceryl monostearate, 0.5 parts by weight of pyruvic acid, 0.5 parts by weight of behenyl alcohol, avocado oil 1 part by weight, 0.05 part by weight of any one of compounds 1 to 5 obtained by the method of Experiment 1, an appropriate amount of vitamin E and an antiseptic is mixed, dissolved by heating according to a conventional method, and 1 part by weight of L-sodium lactate 1, 5 parts by weight of 1,3-butylene glycol, 0.1 part by weight of carboxy polymer and 85.3 parts by weight of purified water were added and emulsified with a homogenizer. Five types of emulsions were obtained.

  Since it contains an antioxidant, the apoptosis-controlling action of compounds 1 to 5 is unlikely to decrease, and this product having a remarkable sunscreen action and skin beautifying action is useful as a cosmetic.

<tablet>
0.5 parts by weight of any one of compounds 1 to 5 obtained by the method of Experiment 1 in 10 parts by weight of L-ascorbic acid, 19 parts by weight of crystalline α-maltose powder and α-glycosyl rutin “αG rutin from Toyo Seika Co., Ltd. After mixing 1 part by weight uniformly, tableting was carried out by a conventional method to obtain 5 types of tablets each containing compounds 1 to 5.

  Because it contains an antioxidant, the apoptosis-regulating action of compounds 1 to 5 is difficult to decrease, easy to take, and stable. This product is useful for the treatment / prevention of various diseases caused by abnormal apoptosis, health recovery, and health maintenance. Useful.

<Liquid>
Sodium chloride 6 parts by weight, potassium chloride 0.3 parts by weight, calcium chloride 0.2 parts by weight, sodium lactate 3.1 parts by weight, maltose 45 parts by weight, any one of compounds 1 to 5 obtained by the method of Experiment 1 and L -0.5 parts by weight of each ascorbic acid was dissolved in 1,000 parts by weight of a 2% (v / v) aqueous ethanol solution, subjected to microfiltration according to a conventional method, and 25 ml each was filled in a sterilized plastic container. Five types of solutions containing 5 to 5 were obtained.

  Since it contains an antioxidant, the apoptosis-regulating action of compounds 1 to 5 is difficult to decrease, and this product, which also has caloric supplementation and mineral supplementation, is used to promote treatment of various diseases caused by abnormal apoptosis and to restore health. It is useful as an injection.

  As explained above, the present invention relates to 3- [4-hydroxy-3,5-bis (3-methyl-2-butenyl) phenyl] -2-propenoic acid and physiologically acceptable salts thereof in humans. This is based on the unique finding that it exerts a remarkable apoptosis regulating action. These compounds are originally isolated from the natural world and have higher safety than those artificially created. Therefore, the apoptosis regulator of the present invention comprising these compounds as active ingredients is in the form of food, cosmetics, pharmaceuticals, etc. to treat / prevent human diseases involving the abnormal apoptosis of cells, restore health, There are practical benefits that can be safely and regularly used for maintenance. In addition, the apoptosis regulator of the present invention containing an antioxidant has a feature that the apoptosis-regulating action of the active ingredient is hardly lowered and can be effectively used for a long time.

It is the photograph which image | photographed the human gastric cancer cell cultured in the presence of 3- [4-hydroxy-3,5-bis (3-methyl-2-butenyl) phenyl] -2-propenoate under an electron microscope. (Magnification 7,000 times). A photograph of normal human skin cells cultured in the presence of 3- [4-hydroxy-3,5-bis (3-methyl-2-butenyl) phenyl] -2-propenoate taken under an electron microscope. Yes (5,000 times magnification). It is the photograph which image | photographed the human leukemia cultured in the presence of 3- [4-hydroxy-3,5-bis (3-methyl-2-butenyl) phenyl] -2-propenoate under a cell electron microscope. (10,000 times magnification). DNA was extracted from human leukemia cells cultured in the presence of 3- [4-hydroxy-3,5-bis (3-methyl-2-butenyl) phenyl] -2-propenoate, and this was extracted from agarose gel electricity. It is the photograph which image | photographed the gel electrophoresis image obtained by electrophoresis.

Claims (1)

  The physiologically acceptable sodium, calcium, magnesium or potassium salt of 3- [4-hydroxy-3,5-bis (3-methyl-2-butenyl) phenyl] -2-propenoic acid is 0.01 An apoptosis promoter that promotes apoptosis of abnormal cells and that does not substantially affect normal apoptosis of normal cells, comprising at least 10% by weight and comprising 0.01 to 10% by weight of an antioxidant.