KR20030062530A - Composition containing an wogonin derivatives for protecting brain cells and treating brain stroke - Google Patents

Abstract

PURPOSE: A composition containing wogonin or a derivative thereof exhibiting preventive and therapeutic effect on ischemic stroke caused by brain cells is provided. Therefore, the composition can be effectively used as a drug and a food supplementary food for protecting the brain. CONSTITUTION: A pharmaceutical composition for treatment of stroke contains 0.5 to 50% by weight of wogonin of the formula(1) or a derivative thereof as an effective component and a pharmaceutically acceptable carrier, based on the total weight of the composition. In formula, the substitutional group of R1, R4 and R5 is each independently H, halogen, C1-3 lower alkyl or lower alkoxy, lower alkyl amine or benzyl; and the substitutional group of R2 and R3 is each independently H, C1-3 lower alkyl or benzyl. The lower alcohol is methyl, ethyl or propyl, and the lower alkoxy is methoxy, ethoxy, propoxy.

Description

COMPOSITION CONTAINING AN WOGONIN DERIVATIVES FOR PROTECTING BRAIN CELLS AND TREATING BRAIN STROKE}

The present invention relates to a composition for protecting and treating brain cells comprising ugonin, that is, derivatives of 5,7-dihydroxy-8-methoxy-2-phenyl-chromen-4-one.

Stroke is divided into two main categories: ischemic stroke, which is caused by an ischemic stroke of the brain tissue due to a decrease or blocking of blood supply to the brain tissue, and a hemorrhagic stroke that causes blood vessels to bleed into the brain tissue. Ischemic stroke accounts for about 80% of all stroke patients, so ischemic stroke is a serious disease. Unlike the existing theories of the immune privileged organs, the nervous system has an inflammatory response in the central nervous system in the case of neurological diseases such as multiple sclerosis, trauma, Alzheimer's disease, cerebral ischemia and various brain injuries. It is reported that this is one of the major causes of neurodegeneration.

Cerebral nerve cell damage caused by stroke may be explained by the release of excessive excitatory neurotransmitters, the production of free radicals, inhibition of protein synthesis, abnormal gene expression, and activation of immune responses. Due to the complexity of cell damage mechanisms, there are no therapeutics that can protect the damage of neurons caused by stroke.

Inhibition of cyclooxygenase-2 (COX-2) in cerebral ischemia has been reported to have neuroprotective effects on the inhibition of production of PGE ₂ and consequent inhibition of glutamate, and many patients with arthritis or pain have already reported COX- Because of the use of inhibitors, it has been reported that epidemiological investigations of stroke incidence in these patients could be new targets for the prevention and treatment of stroke in the future [Iadecola et al. See Iadecola, C. et al., PNAS. , 30 p1294-1299, 2001.

On the other hand, inflammatory reactions occur in the central nervous system, which has been reported to be one of the main causes of neurodegeneration. Inflammatory response after cerebral ischemia is expected to be the target of new cerebrovascular ischemic agents. It is thought to consist of drugs that inhibit enzymes or inhibit the deposition of neutrophils (Dirnagl, U. et al; TINS ., 22 , pp391-397, 1999).

In addition, inducible nitric oxide synthase (iNOS), which can release NO from neutrophils penetrated into brain tissue, is induced, and NO is released from induced iNOS, which is known to cause brain cell damage. Neurotoxicity has also been reported to be via NO, and development of neuroprotective agents related to iNOS expression inhibition is expected (Vaughan CJ, Delanty N., Stroke 30: 1969-73, 1999).

Ugonin of the present invention is a flavone-based compound that is isolated from Scutellariae radix, Wakabayashi and Yasui are Ugonin by lipopolysaccharide in macrophage (lipopolysaccharide) It is reported to inhibit the production of induced prostaglandin E2 (Wakabayashi, I. and Yasui, K., Eur. J. Pharmacol. , 406 , pp 477-481, 2000). Gao et al. Reported that ugonin and baicalin contained in Scutellariae radix have antioxidant activity and may be useful for the treatment of diseases caused by oxidative stress (Gao). et al., Pharmacol Res. 43 : pp 173-8, 2001). Shieh et al reported that ugonin, baikalline, and baicalein would be effective in brain tumors, suggesting that it was due to the inhibitory activity of xanthine oxidase, which is involved in the production of oxides (Shieh). et al., Anticancer Res. , 20 : pp 2861-5, 2000). Chang et al. Reported that among the various flavonoid compounds such as quercetin, ugonin, and bikallein, the most effective inhibitor of xanthine oxidase is Baikallein, and these compounds are known as hepatitis and brain tumors. It is reported to be useful (Chang et al., Anticancer Res., 13 : 2165-70, 1993). Kim et al. Reported that they inhibit cell death by inhibiting NO production using C6 rat glial cells derived from ugonine-activated glial cells (Kim et al., Neurosci Lett). ., 309 : pp67-71, 2001). Hui et al. Also suggest the possibility of ugonin as an anticonvulsant because of its strong binding ability to the benzodiazepine receptor (Hui et al., Planta Med., 66 : pp91-3, 2000).

However, all of these documents are the results of experiments using cells, not reports using animals. It is very effective at the cellular level, but in vivo it is not possible to develop it as a medicine without considering the variable blood-brain barrier. In other words, to protect brain damage caused by cerebral ischemia, the drug must move through the blood-brain-barrier to brain cells to exert an effect. Therefore, in vivo experiments are indispensable if you want to see the effects of drugs that have brain injury protection. However, none of the above documents specifically suggests the protective effect of ugnin on brain injury by cerebral ischemia through in vivo experiments.

Meanwhile, Korean Unexamined Patent Publication No. 2001-081188 (published on Aug. 29, 2001) discloses a golden extract having a neuroprotective effect and a pharmaceutical preparation containing the same, and Park et al. The anti-inflammatory action of ugonin is reported (Park et al., Eur J Pharmacol., 425 : pp153-7, 2001).

However, there is no teaching or suggestion in the above documents that ugonin and its derivatives isolated from gold may exhibit protective effects of brain injury by cerebral ischemia.

It is an object of the present invention to provide a ugonin-containing pharmaceutical composition and a dietary supplement showing a protective effect on brain cell transmission and a stroke prevention and treatment effect.

1 is a diagram showing the results of experiments on the inhibitory effect of TNF-α by ugonin in BV2 microglial cell line,

2 is a diagram showing the results of experiments on the effect of inhibiting the nitrite (Nitrite) production by the ugonin in BV2 microgreal cell line.

In accordance with the above object, the present invention provides a composition for protecting and preventing brain cells and stroke comprising a ugonin or a derivative thereof represented by the following formula (1).

Where

R ₁ , R ₄ and R ₅ substituents are each independently hydrogen, a halogen group, a C ₁ to C ₃ lower alkyl or lower alkoxy group, a lower alkyl amine group or a benzyl group,

R ₂ and R ₃ substituents are each independently hydrogen, C ₁ to C ₃ lower alkyl or benzyl groups.

Lower alkyl group in the present invention means methyl, ethyl, propyl group, lower alkoxy group means methoxy, ethoxy, propoxy group, halogen group means atoms such as fluorine, chlorine, bromine, iodine.

The brain cell protection and memory enhancing composition of the present invention, the compound of Formula 1 comprises about 0.5 to 50% by weight relative to the total weight of the composition.

Ugonin derivatives having brain cell protection and stroke prevention and treatment of the present invention can be synthesized through the synthesis and fractionation of conventional substituents. (Herbert O. House: Modern Synthetic Reactions, 2nd Ed., The Benjamin / Cummings Publishing Company, 1972)

The effect on the prevention and treatment of stroke of the composition of the present invention can be confirmed by the effect of inhibiting ischemic stroke of ugonin and its derivatives in vivo using a midbrain aortic occlusion model in which the test method has already been established.

The method involves inserting nylon filament into the internal carotid artery to occlude the middle cerebral aorta and then reperfusion by removing the filament after 120 minutes. Oral administration of ugonin and its derivatives to oral administration for ischemic stroke To see the effect.

Compositions comprising the ugonin of the present invention and derivatives thereof may further comprise suitable carriers, excipients and diluents commonly used in the manufacture of pharmaceutical compositions.

Carriers, excipients and diluents that may be included in the composition comprising the ugonin and derivatives of the present invention include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate , Gelatin, calcium phosphate, calcium silicate, cellulose, methyl cellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil.

Compositions comprising ugonin and its derivatives according to the present invention may be in the form of powders, tablets, capsules, suspensions, emulsions, syrups, aerosols and the like, oral preparations, suppositories, and sterile injectable solutions, respectively, according to conventional methods. Can be formulated and used.

The amount of ugonine derivatives may vary depending on the age, sex, and weight of the patient, but may be administered once or several times in an amount of 0.1 to 500 mg / kg body weight per day. The dosage of the ugonin derivatives can be increased or decreased depending on the route of administration, the severity of the disease, sex, weight, age and the like. Therefore, the above dosage does not limit the scope of the present invention in any aspect.

Ugonin or a derivative comprising the composition according to the present invention can be used in various formulations, such as drugs, foods and beverages for the protection of brain cells and memory enhancement in such a formulation. Foods to which ugonine derivatives can be added include, for example, various foods, beverages, gums, teas, vitamin complexes, and dietary supplements.

In addition, the ugonin and its derivatives according to the present invention may be added to food or beverage for the purpose of protecting brain cells and preventing stroke. In this case, the amount of the ugonin derivative in the food or beverage is generally 0.1 to 15% by weight, preferably 1 to 10% by weight of the total food weight in the case of the health food composition of the present invention, health drinks The composition may be added at a ratio of 1 to 30 g, preferably 3 to 10 g, based on 100 ml.

The health beverage composition of the present invention has no particular limitation on the liquid component except for containing the ugolinine derivative as an essential ingredient in the indicated ratio, and may contain various flavors or natural carbohydrates as additional ingredients, such as ordinary drinks. Examples of the above-mentioned natural carbohydrates include monosaccharides such as glucose, fructose and the like; Disaccharides such as maltose, sucrose and the like; And conventional sugars such as polysaccharides such as dextrin, cyclodextrin, and sugar alcohols such as xylitol, sorbitol, and erythritol. As flavoring agents other than those mentioned above, natural flavoring agents (tauumatin, stevia extract (e.g., rebaudioside A, glycyrrhizin, etc.), and synthetic flavoring agents (saccharin, aspartame, etc.) can be advantageously used. The ratio of said natural carbohydrate is generally about 1-20 g, preferably about 5-12 g per 100 ml of the composition of the present invention.

In addition to the above, the composition of the present invention includes various nutrients, vitamins, minerals (electrolytes), flavors such as synthetic flavors and natural flavors, coloring and neutralizing agents (such as cheese and chocolate), pectic acid and salts thereof, alginic acid and salts thereof. , Organic acids, protective colloid thickeners, pH adjusting agents, stabilizers, preservatives, glycerin, alcohols, carbonation agents used in carbonated beverages, and the like. The compositions of the present invention may also contain pulp for the production of natural fruit juices and fruit juice beverages and vegetable beverages. These components can be used independently or in combination. The proportion of such additives is not so critical but is generally selected from the range of 0 to about 20 parts by weight per 100 parts by weight of the composition of the present invention.

The present invention is described in more detail based on the following examples, although the present invention is not limited thereto.

Reference Example 1. 7- O Preparation of Benzyl Crycine

After dissolving 10.2 g (40 mmole) of chrysine in 200 ml of anhydrous acetone, 12.5 g (90 mmole) of anhydrous potassium carbonate and 5 ml (42 mmole) of benzyl bromide were added and reacted under reflux for 5 hours. After cooling the reaction solution, the solution was added to 800 ml of 1N dilute aqueous hydrochloric acid solution, and the resulting white white crystals were filtered and dried in air at about 60 ^o C to obtain 13.8 g of 7- O -benzyl chrysine, which was not subjected to separate purification. It was used as it is for the next reaction.

¹ H-NMR (200 MHz, CDCl ₃ ): δ 5.15 (s, 2H), 6.46 (d, 1H, J = 2.4 Hz), 6.59 (d, 1H, J = 2.4 Hz), 6.68 (s, 1H) , 7.38-7.55 (m, 8H), 7.86-7.91 (m, 2H), 12.74 (s, 1H)

Reference Example 2. 7- O Preparation of -benzyl-5,7-dihydroxy-2-phenyl-chromen-4-one

7 g of 7- O -benzylcrisine obtained in Reference Example 1 was dissolved by adding an aqueous solution in which 6.3 g of potassium hydroxide was dissolved in 200 ml of pyridine and 120 ml of water, followed by dissolving 10.5 g of K ₂ S ₂ O _{8 in} 300 ml of water. It was added slowly at room temperature over 2 hours, and reacted for 24 hours. The light brown reaction solution was added to 1000 ml of water, and the pH was adjusted to pH 6 by adding 6N aqueous hydrochloric acid solution to recover the unreacted pale yellow 7- O -benzylcristine precipitate, which was then precipitated. Was added, heated at about 90 ^o C for 2 hours, and allowed to stand at room temperature to gradually precipitate crystals. The slightly yellow crude product was purified again by recrystallization and 2.4 g of 7- O -benzyl-5,7-dihydroxy-2 was added. The -phenyl-chromen-4-one product was obtained.

¹ H-NMR (200 MHz, CDCl ₃ ): δ 5.22 (s, 2H), 6.53 (s, 1H), 6.71 (s, 1H), 7.43-7.58 (m, 8H), 7.94-7.98 (m, 2H ), 12.30 (s, 1 H)

Reference Example 3. Preparation of 7-benzyloxy-5-hydroxy-8-methoxy-2-phenyl-chromen-4-one

After dissolving 3.2 g of 7- O -benzyl-5,7-dihydroxy-2-phenyl-chromen-4-one obtained in Reference Example 2 in 150 ml of acetone, 8 g of potassium carbonate and 0.4 ml of dimethyl sulfate were added thereto for 6 hours. It was refluxed. After cooling the reaction solution, the resulting dark yellow solid was added to 800 ml of 12% aqueous hydrochloric acid solution. The resultant mixture was separated by filtration. Recrystallized with methanol, 2.2 g of a slightly yellow solid, 7-benzyloxy-5-hydroxy-8-methoxy- 2-phenyl-chromen-4-one was obtained.

¹ H-NMR (200 MHz, CDCl ₃ ): δ 3.97 (s, 3H), 5.23 (s, 2H), 6.49 (s, 1H), 6.69 (s, 1H), 7.38-7.56 (m, 8H), 7.94-7.98 (m, 2H), 12.54 (s, 1H)

Example 1 Preparation of 5,7-Dihydroxy-8-methoxy-2-phenyl-chromen-4-one (ugonine)

In Reference Example 3, 1.9 g of 7-benzyloxy-5-hydroxy-8-methoxy-2-phenyl-chromen-4-one was dissolved in 20 ml of concentrated hydrochloric acid and 66 ml of vinegar acid, and heated to reflux for 2 hours. The precipitated solid was filtered during the reaction, washed with water and recrystallized to obtain 1.1 g of pale yellow ugonin.

¹ H-NMR (200 MHz, DMSO-d ₆ ): δ 3.87 (s, 3H), 6.33 (s, 1H), 7.03 (s, 1H), 7.60-7.65 (m, 3H), 8.07-8.11 (m , 2H), 10.85 (s, 1H), 12.53 (s, 1H)

¹³ C-NMR (50 MHz, DMSO-d ₆ ): δ 60.9, 99.0, 105.0, 126.2, 127.7, 129.2, 130.8, 132.0, 149.6, 156.2, 157.3, 163.0, 182.1

mp = 195-197 ^o C

Experimental Example 1 Protective Effect of Ugonin and Its Derivatives on Ischemic Stroke

The ugonin and ugonin derivatives obtained in Example 1 were administered at 1, 3, 10, 50 mg / kg and reperfused about 5 minutes before the start of reperfusion in a rat ischemic animal model using rats (Biogenomeics, Korea). Approximately 24 hours after the first operation, the animal is killed and the brain is extracted, and a brain slice is made 2 mm thick using a brain matrix, and then 2, 3, 5-triphenyltetrazolinum After staining using Chloride (TTC) staining, the cerebral infarction site was analyzed using an image analysis system. As shown in Table 1 below, ugonin and its derivatives exhibited the maximum brain injury protection effect by cerebral ischemia at 10 mg / kg, and it was known to be effective for cerebral ischemia and used as a positive control. ) And it was confirmed that the effect is superior to Baikalein (baicalein).

The protective effect of the following Table 1 is converted in% of the degree of protection for the vehicle (physiological saline or 5% carboxymethylcellulose) administration group.

Cerebral Ischemia Protective Effect of Various Ugonin Derivatives Treatment group Protective effect (%) Ebceline (30 mg / kg x 1) 45.2% Bicalane (50 mg / kg x 2 times) 41.1% Ugonin (1 mg / kg x 1) 27.6% Ugonin (5 mg / kg x 1) 49.8% Ugonin (10 mg / kg x 1) 68.5% Ugonin (50 mg / kg x 1) 71.2%

Experimental Example 2 Free Radical Scavenging Effect of Ugonin Derivatives

DPPH (1,1-diphenyl-2-picrylhydrazyl radical) scavenging function was used as an index of free radical scavenging activity. Since DPPH itself is a free radical, it is known to have a function of removing purple color of DPPH in the case of a compound having antioxidant activity. Experimental methods have already used a known method described in several documents (Vivot, E. et al., J. Ethnopharmacology, 76 pp65-71, 2001), the results are shown in Table 2 below. It was also confirmed that the DPPH scavenging activity of the wogoin derivatives was excellent.

DPPH Scavenging Activity Experiment Results Sample group (1 mg / ml) effect (%) Evceline 5.4% Baikallein 92.2% Ugonin 91.2%

Experimental Example 3. Scavenging function of peroxynitrite of ugonine derivatives

The purpose of this study is to determine how much of the ugonin derivatives can scaveng the peroxynitrite produced by SIN-1 (3-morpholinosydnonimine) to determine the mechanism of action of ugonin and its derivatives. First, 200 μM of SIN-1 and various concentrations (0.05-300 μg / ml) of ugonin and its derivatives were dissolved in PBS and incubated at 37 ° C. for 20 minutes. After incubation, 100 μM of dihydrorhodamine-123 (DHR123), which is oxidized by peroxynitrite and generates fluorescence, was added to the culture medium, and then incubated again at 37 ° C. for 30 minutes to give 536 nm (emission), Fluorescence was measured at 500 nm (excitation) to determine the degree of scavenging action of peroxynitrite of the ugonine and its derivatives. The results are shown in Table 3 below. As a result, it was confirmed that the ugonin and its derivatives were very excellent in the scavenging activity of the peroxynitrite produced by SIN-1 (IC ₅₀ : 200 µg / ml).

Peroxynitrite Scavenging Activity of Ugonin and Its Derivatives Concentration (㎍ / ml) Oxidation Rate of DHR123 (%) 10 minutes 20 minutes 40 mins 60 mins 0 100 100 100 100 3 97 98 101 102 10 99 101 98 99 30 96 97 99 95 100 87 88 90 86 300 32 41 28 30 1000 6 4 7 5

Experimental Example 4 Inhibition of Nitrite Production by Activated Microglial Cells

Dispense 100 micrograms of mouse microglial cell-based BV2 cells (Weill Medical College of Cornell University, USA) into 24 well plates at room temperature per L, incubate overnight in medium, and then test for 100 ng / ml of LPS and test drugs. After incubation for 24 hours and the amount of nitrite released into the medium using Grease's reagent (Choi, JJ and Kim, WK, J. Neurosci. Res., 54 : 870-875, 1998). As shown in FIG. 1, ugonin reduced the amount of NO released by LPS (100 ng / ml) to about 1/2 at a concentration of 60 μM and reduced to about 1/8 at a concentration of 100 μM. I was. The NO inhibitory effect of ugonin showed similar results in microglia cells of primary culture rats.

Experimental Example 5: Inhibition of TNF-α Production by Activated Microglial Cells

Treating BV2 mouse microglia with LPS and test drugs in the same manner as in Experimental Example 4 and releasing the TNF-α released into the medium after 6 or 24 hours of culturing with the TNF-α ELISA kit (DIACLONE Research, USA). It measured using.

As shown in FIG. 2, ugonin reduced TNF-α liberated by LPS (100 ng / ml) to about 1/2 at a concentration of 10 μM and 100 μM to about 1/3 to 1/4 I was. The degree of reduction was the same at 6 hours and 24 hours. Ugonin's TNF-α free inhibitory activity showed similar results in primary cultured rat microglial cells.

Experimental Example 6. Acute toxicity test of ugonin and its derivatives using mouse

Mice were used for acute toxicity testing of various ugonin derivatives. As a result of the acute toxicity test, no mortality was observed at 330 mg / kg of ugonine derivatives, and no significant abnormalities were found in weight gain and feed intake. Therefore, it was found that the ugonine derivative is a safe drug.

An example of the preparation of the pharmaceutical composition is exemplified below, which is intended to explain in detail rather than limit the present invention thereto.

Formulation Example 1 Preparation of Powder

According to the preparation method of powder in the pharmacopeia formulation, it is prepared in the following component content per packet.

Ugonin Powder ・ ・ ・ ・ ・ ・ ・ ・ ・ 300 mg

Lactose · ・ ・ ・ ・ ・ ・ ・ ・ ・ ・ ・ ・ ・ ・ ・ 100 mg

Talc 10 mg

Formulation Example 2 Preparation of Tablet

According to the preparation method of tablets in the pharmacopeia formulation, it is prepared in the following component content per tablet.

Ugonin Powder ・ ・ ・ ・ ・ ・ ・ ・ ・ 300 mg

Corn starch ㆍ ・ ・ ・ ・ ・ ・ ・ ・ ・ ・ ・ ・ ・ ・ ・ ・ ・ ・ 100 mg

Lactose · ・ ・ ・ ・ ・ ・ ・ ・ ・ ・ ・ ・ ・ ・ ・ ・ 100 mg

Magnesium Stearate 2 mg

Formulation Example 3 Preparation of Capsule

According to the preparation method of capsules in the pharmacopeia formulation, it is prepared in the following component content per capsule.

Ugonin Powder ・ ・ ・ ・ ・ ・ ・ ・ ・ 300 mg

Corn starch ・ ・ ・ ・ ・ ・ ・ ・ ・ ・ ・ ・ ・ ・ ・ ・ 100 mg

Lactose ····················· 100 mg

Magnesium stearate 2 mg

Formulation Example 4 Preparation of Injection

According to the preparation method of injection in the pharmacopeia formulation, it is prepared in the following component content per ampoules (2 ml).

Ugonin Powder ・ ・ ・ ・ ・ ・ ・ ・ ・ 300 mg

Sterile distilled water for injection ㆍ ·············

pH regulator ㆍ ··················

Formulation Example 5 Preparation of Liquid

According to the preparation method of the liquid formulation in the Pharmacopoeia General Formulation, it is prepared in the following component content per 100 ml of the liquid formulation.

Ugonin Powder ... ・ ・ ・ ・ ・ ・ ・ ・ 1 g

Heterosaccharides ················· 10 g

Mannitol ···················· 5 g

Purified water ㆍ ···················

In addition, to prepare a healthy beverage containing ugonin in the following manner.

Add 0.05% to 25% of ugonine powder, 5% to 10% sugar, 0.05% to 0.3% citric acid, 0.005% to 0.02% caramel, and 0.1% to 1% vitamin C and mix it with 79% to 94% purified water to make syrup. In addition, the syrup was sterilized at 85 to 98 ° C. for 20 to 180 seconds, mixed with cooling water at a ratio of 1: 4, and then a carbonated beverage containing ugonine powder was prepared by injecting carbon dioxide gas at 0.5 to 0.82%.

As a subsidiary ingredient, a healthy beverage composed of liquid fructose (0.5%), oligosaccharide (2%), sugar (2%), salt (0.5%) and water (75%) may be prepared.

Although the composition ratio is a composition suitable for a preferred beverage in a preferred embodiment, the compounding ratio may be arbitrarily modified according to regional and ethnic preferences such as demand hierarchy, demand country, and use purpose.

The composition containing ugonin and its derivatives can be usefully used as a brain protection agent for treating brain injury since it exhibits a prophylactic and therapeutic effect against ischemic stroke due to damage of cerebral nerve cells.

Claims (3)

A pharmaceutical composition for treating stroke, comprising a ugonine or a derivative thereof represented by Formula 1 as an active ingredient and a conventional pharmaceutically acceptable carrier: Formula 1 Where R ₁ , R ₄ and R ₅ substituents are each independently hydrogen, a halogen group, a C ₁ to C ₃ lower alkyl or lower alkoxy group, a lower alkyl amine group or a benzyl group, R ₂ and R ₃ substituents are each independently hydrogen, C ₁ to C ₃ lower alkyl or benzyl groups. The method of claim 1, Pharmaceutical composition characterized in that the stroke is an ischemic stroke A dietary supplement comprising a ugonin or a derivative thereof and a foodstuff acceptable food supplement additive that have a protective effect on brain cells and prevent stroke.