KR101060909B1 - Composition comprising plantain extract comprising brain neuronal cell protective material - Google Patents

Abstract

The present invention relates to a composition for the prevention or treatment of degenerative neurological diseases comprising a plantain extract comprising a brain neuronal cell protective material. More specifically, it contains 4-vinyl-2-methoxyphenol as a brain neuronal cell protective agent, reduces oxidative stress induction and increases cell viability, Alternative behavior and step through latency The present invention relates to a composition for the prevention or treatment of degenerative neurological diseases harmless to the human body including the plantain extract that maintains memory learning ability in a passive avoidance test.

Plantain extract, amyloid beta peptide-induced oxidative stress, degenerative neurological disease

Description

Composition comprising Plantago asiatica extract which contains inhibitor aganist amyloid β peptide-induced oxidative stress}

The present invention relates to a composition for degenerative neurological diseases comprising a plantain extract comprising a brain neuronal cell protective material, and more particularly, to inhibit brain neurotoxicity caused by amyloid beta peptide-induced oxidative stress. It relates to a composition comprising a plantain extract comprising an active ingredient having.

Recently, due to prolonged life expectancy and decrease in fertility rate, the elderly population aged 65 or older has already entered the aging society with 7.2% of the total population as of 2000.In this trend, 14.4% in 2019, and 2026 It is expected to reach an ultra-old society at 20.0%. Due to such changes in social conditions, the number of deaths from infectious diseases, which are the major causes of death, has been drastically decreasing, while the number of deaths from cancer, cerebrovascular diseases, and metabolic syndrome is steadily increasing. In particular, as life expectancy increases and the aging of society progresses, interest in aging and diseases related to aging, in particular, neurodegenerative diseases related to brain nerve cell damage, are increasing. In the United States, Alzheimer's disease (AD) is the fourth leading cause of death in all populations, after cardiovascular disease, malignancy and stroke, and in terms of treatment costs, third place after cancer and heart disease. As it occupies the above, the socio-economic impact seems to be significant.

The hallmark of Alzheimer's disease is the formation of neurofibrillary tangles and amyloid plaques, which are confirmed by cytopathologic examination of the patient's postmortem brain, which results in excessive phosphorylation of the tau protein and amyloid beta peptides , Hereinafter referred to as 'Aβ').

In patients with hereditary AD, mutations in the amyloid precursor protein (hereinafter referred to as 'APP') or presenilin genes increase A β and all progress to AD. Known as a factor. Aβ is a 40- or 42-protein protein produced by cleaving the precursor APP, beta-site APP-cleaving enzyme 1 (BACE1), and cutting the resulting C-terminal gamma secretase, including presenilin. The pieces are formed. Inactivation and necrosis of the cranial nerve are thought to be cytotoxic by Aβ, an abnormal metabolite of APP. Its neurotoxicity begins with brain nerve cell destruction caused by oxidative stress. The cause of neural cell necrosis in AD is that the production of oxygen free radicals and oxidative injury such as reactive oxygen species (ROS) are associated with brain nerve cell destruction and pathogenesis. It is estimated to be.

Recent AD studies have focused on the destruction of brain cells by Aβ and the reduction of levels of the neurotransmitter acetylcholine (ACh) due to their protein structure, physiology and accumulation in the brain. To date, the main targets for AD drug development have been FDA-approved by cholinesterase inhibitors (Aricept, Exelon, Reminyl, etc.) and glutamate antagonist Memantine, which target cholinergic neurons beyond neurotransmitters present in disease. Is currently on the market. However, since these drugs only temporarily relieve symptoms, there is an urgent need for drug development techniques that treat the underlying disease or inhibit the progression of the disease itself.

On the other hand, plantain (Plantago asiatica ) is a perennial herb belonging to Plantaginaceae, which is native to East Asia and Central Asia such as Korea, China, and Japan. The dried outposts of flowers are called chajeoncho and seeds as chajeon, which is widely used in nephritis, cystitis and urethritis as a powerful diuretic, and minerals, proteins, vitamins and polysaccharides It contains a lot of these ingredients and is known to be involved in anti-inflammatory action.

The main bioactive components of plantain are iridoid glycosides, acubin and plantaglucoside, various sterols, adenine, choline and mucilage And various fatty acids. The main research related to plantain is to isolate and confirm aucubin from H ₂ O fraction as the main physiologically active component of plantain acting as interpolation agent, and also to prevent antibacterial, bile secretion, laxative action and hepatotoxicity Actions and blood pressure lowering actions have been reported. In addition, there have been studies on the anticancer effect using various kinds of herbal medicines including plantain, and on the effect of the generation of free radicals in liver and the regulation of antioxidant enzyme system, but the study on the effect of plantain extract on the toxicity of brain neurons by amyloid beta-induced oxidative stress There were very few results.

Therefore, the present inventors searched for phytochemicals for the prevention and treatment of brain diseases including brain cell necrosis, which are increasingly increased in aging diseases, and edible in preparing them as raw materials for medicine, food medicine and health functional food. Plant resources were searched to secure the safety of the final material and to increase the probability of practical use as a functional food material. In search of dozens of domestic food and medicinal plants, we searched for natural new materials that protect brain cells from oxidative stress caused by Aβ. The brain nerve cell damage protection material by the confirmation was confirmed, and this invention was completed.

An object of the present invention is to provide a pharmaceutical composition for the prevention or treatment of degenerative neurological diseases, including the plantain extract.

Another object of the present invention is to provide a pharmaceutical composition for preventing or treating neurodegenerative diseases including 4-vinyl-2-methoxyphenol.

Still another object of the present invention is to provide a dietary supplement for preventing or improving degenerative neurological diseases including plantain extract or 4-vinyl-2-methoxyphenol.

In one embodiment, the present invention relates to a pharmaceutical composition for preventing or treating neurodegenerative diseases comprising plantain extract or 4-vinyl-2-methoxyphenol.

Hereinafter, the present invention will be described in detail.

In the present invention, degenerative neurological disease refers to a term that generically refers to various diseases related to nerves, in particular, cranial nerves. Preferably, the neurodegenerative disease refers to a disease causing brain nerve cell damage caused by amyloid beta peptide-induced oxidative stress, specifically, Alzheimer's disease, dementia, amyotrophic laternal sclerosis, and Parkinson's disease. (Parkinson's disease), myocardial infacrtion or Huntington's disease, more preferably Alzheimer's disease.

Specifically, amyloid beta was found to accumulate in the mitochondria of neurons of Alzheimer's disease model, and the production of free radicals in mitochondria has been reported to cause oxidative damage. It has been reported to be associated with the development of Alzheimer's disease (Manczak et. al ., Hum Mol Genet. 15 (9): 1437-49); Reddy, J Neurochem . 96 (1): 1-13 (2006).

Plantain extract included in the composition according to the present invention can be prepared using any extraction method well known in the art without limitation. Therefore, conditions such as extraction time, solvent and temperature can be appropriately adjusted by those skilled in the art. Preferably it is extracted using ethanol.

The plantain extract included in such a composition according to the present invention has a protective activity against nerve cell damage caused by amyloid beta peptide-induced oxidative stress.

In In vitro , the plantain ethanol extract was fractionated in the order of hexane, chloroform, ethyl acetate according to the polarity, and then the oxidative stress induction and the cell activity (viability) were measured through the MTT reduction assay. Neuroprotective activity was confirmed.

Also in vivo In mice, alternative behavior and step through latency were passive in the group of 400 mg / kg and 800 mg / kg doses of plantain extract after amyloid beta was injected into mice. In the avoidance test, the effect was superior to the dementia group.

The active ingredient having a protective activity against amyloid beta-peptide-induced oxidative stress-induced nerve cell damage contained in the plantain extract is 4-vinyl-2-methoxyphenol, which may also be included in the composition according to the present invention. . 4-vinyl-2-methoxyphenol is not limited thereto, but is prepared by extraction and separation in the following manner.

First plantain (Plantago adding ethanol to the plantain obtained by pulverizing asiatica ), to prepare a plantain ethanol extract, fractionating the ethanol extract using an organic solvent having a different polarity, and obtaining the isolate; Obtaining the brain damage inhibiting active ingredient using an open silica gel column of the material obtained in the above step; And separating the active fractions obtained in the previous step using HPLC using a reversed phase column. Schematic of this manufacturing method is as follows.

[Scheme 1]

Methanol, ethanol, isopropyl alcohol, butanol, and the like may be used as the extract, preferably ethanol. In addition, hexane, chloroform and ethyl acetate are used as organic solvents having different polarities.

On the other hand, degenerative neurological disease prevention or treatment composition according to the present invention may comprise a pharmaceutically acceptable carrier in addition to plantain extract or 4-vinyl-2-methoxyphenol. Furthermore, the composition of the present invention may contain one or more known active ingredients having the effect of neuronal cell protection and neurotoxicity inhibition or prevention together with plantain extract or 4-vinyl-2-methoxyphenol.

The composition of the present invention can be prepared by including one or more pharmaceutically acceptable carriers in addition to the above-described active ingredients for administration. Pharmaceutically acceptable carriers may be used in combination with saline, sterile water, Ringer's solution, buffered saline, dextrose solution, maltodextrin solution, glycerol, ethanol, and one or more of these components, as necessary. And other conventional additives such as bacteriostatic agents. In addition, diluents, dispersants, surfactants, binders, and lubricants may be additionally added to formulate into injectable solutions, pills, capsules, granules or tablets such as aqueous solutions, suspensions, emulsions and the like. Furthermore, it may be preferably formulated according to each disease or component by a suitable method in the art or using a method disclosed in Remington's Pharmaceutical Science (Recent Edition), Mack Publishing Company, Easton PA. Examples of acceptable carriers, formulations of pharmaceutical compositions, and methods of preparing these formulations are by known methods.

These pharmaceutical compositions are useful for administering the plantain extract of the present invention to a subject for the treatment of various diseases, including neurodegeneration and / or symptoms associated therewith, as described herein. Plantain extract is provided in an amount effective to treat neurodegenerative diseases in a subject to which the pharmaceutical composition is administered, and the dosage is weight, age, sex, health condition, diet, time of administration, method of administration, excretion rate and disease of the patient. The range varies depending on the severity of the disease. As noted above, one skilled in the art can conveniently determine an appropriate amount.

The compositions of the present invention can be administered orally or parenterally (eg, intravenously, subcutaneously, intraperitoneally, intramuscularly, intravascularly or subcutaneously) according to the desired method, preferably orally or intravenously. to be.

For oral administration, the plantain extract formulation may be provided in capsules, tablets, powders, granules or suspensions. These formulations may contain conventional additives such as lactose, mannitol, corn starch or potato starch. These formulations may also be provided with binders such as crystalline cellulose, cellulose analogs, acacia, corn starch or sodium carboxymethyl cellulose. These formulations may also be provided with dibasic calcium phosphate or anhydrous sodium starch glycolate. Finally, these formulations may be provided with lubricants, such as talc or magnesium stearate.

For parenteral administration, the plantain extract preparation can be mixed with a sterile aqueous solution, preferably with a sterile aqueous solution that is isotonic with the blood of the individual. These formulations can be prepared by dissolving the solid active ingredient in water containing a physiologically compatible substance such as sodium chloride, glycine and the like and having a buffered pH suitable for physiological conditions to produce an aqueous solution and to make the solution sterile. have.

The composition of the present invention can be used alone or in combination with methods using surgery, hormonal therapy, drug therapy and biological response modifiers for the prevention or treatment of degenerative neurological diseases.

As another aspect, the present invention relates to a composition of a dietary supplement for the purpose of preventing or improving degenerative neurological diseases. When the plantain extract of the present invention is used as a food additive, the plantain extract or 4-vinyl-2-methoxyphenol may be added as it is, or used together with other food or food ingredients, and may be appropriately used according to a conventional method. . The mixed amount of the active ingredient can be determined suitably according to the purpose of use (prevention, health or therapeutic treatment).

There is no particular limitation on the kind of food. Examples of the food to which the substance may be added include meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen, other dairy products including noodles, gum, ice cream, various soups, drinks, tea, drink, Alcoholic beverages and vitamin complexes, and the like and include all of the health foods in the conventional sense.

The plantain extract of the present invention is a natural product which can be easily obtained from the surroundings and has little concern about toxicity, side effects, etc., and in particular, the fact that the extract contains a protective substance against nerve cell damage caused by oxidative stress. Newly revealed. Therefore, the plantain extract or the composition containing 4-vinyl-2-methoxyphenol according to the present invention can be usefully used as a pharmaceutical composition and health functional food composition for the prevention or treatment of neurodegenerative diseases, preferably Alzheimer's disease. .

As described above, the plantain extract of the present invention not only reduces oxidative stress and increases cellular activity, which is known to be correlated with degenerative neurological diseases, but also maintains memory learning ability in dementia models and does not cause side effects in the human body. The composition comprising may be usefully used as a pharmaceutical or nutraceutical composition for the prevention and improvement of the disease.

Hereinafter, preferred embodiments of the present invention will be described in order to facilitate understanding of the present invention. However, the following examples are merely provided to more easily understand the present invention, and the contents of the present invention are not limited by the examples.

Oxidative stress was measured by spectrofluorometer using DCF-DA (2 ＇, 7'-dichlorofluorescin diacetate) assay to confirm the inhibitory activity of plantain extract, and cell viability. ) Was measured via MTT reduction assay.

Example 1 confirmed brain neuroprotective activity of ethanol extract of plantain

In order to confirm the neuroprotective activity of cerebral neurons from oxidative stress-induced cerebral neuronal cytotoxicity, planar neuronal necrosis inhibition was measured for each concentration of plantain ethanol extract.

PC12 cells were cultured in RPMI-1640 medium containing antifungal / antibiotic and treated with 100 μM of H ₂ O ₂ to induce oxidative stress when the number of cells reached 10 ⁴ -10 ⁶ cells / ml.

Group C is deionized water treatment group and Group H is H ₂ O ₂ 100 μM treatment group, V was Vit. For 48 hours before H ₂ O ₂ treatment. C pre-incubated with 100 μM, the other groups were H ₂ O ₂ Precultured at different concentrations (0.5-2.0 mg / ml) for 48 hours before treatment. The degree of oxidative injury induction was measured through a spectrofluorometer using a DCF-DA assay to confirm the inhibitory activity of the plantain extract (see FIG. 1). Cell viability was measured via MTT reduction assay.

As shown in Figure 1, it was confirmed that the degree of oxidative stress induced by H ₂ O ₂ is reduced in proportion to the concentration of the ethanol extract, the cell activity (cell viability) compared to the vitamin C pre-treatment group ethanol extract The treatment group showed good activity.

Example 2 Plantain ethanol extract prepared and brain nerve cell protective material separating

1. Fractionation from plantain ethanol extract using organic solvents with different polarities

Selected Plantain ( Plantago Asiatica ) finely pulverized with a hand mixer, 3 kg of plantain is extracted with shaking 5 times the volume of ethanol five times for 24 hours shaking (shaking) and then concentrated under reduced pressure to prepare a plantain ethanol extract. Completely remove ethanol from the extract, and then fractionate three times in the order of hexane, chloroform, and ethyl acetate, depending on the polarity. The degree of oxidative stress and cell viability (%) were determined. Confirmed. Group C is deionized water treatment group and Group H is H ₂ O ₂ 100 μM treatment group, V is H ₂ O ₂ Vit. For 48 hours before treatment. C 100 μM pre-incubated group, the other groups H ₂ O ₂ Different treatments for 48 hours before treatment (H1 is the first layer fractionated with hexanes; H2 is the second layer fractionated with hexanes; H3 is the third layer fractionated with hexanes; C1 is the first layer fractionated with chloroform; C2 Is the second layer fractionated with chloroform; C3 is the third layer fractionated with chloroform; E1 is the first layer fractionated with ethyl acetate; E2 is the second layer fractionated with ethyl acetate; and E3 is fractionated with ethyl acetate The third layer). As shown in FIG. 2, the second fractional layer (E2) and the third fractional layer (E3) of the ethyl acetate layer showed the highest oxidative stress inhibitory activity and the highest cellular activity (see FIG. 2).

2. The separated ethyl acetate fractions were opened on silica gel. Column  Through fractions

The E2 and E3 layers of the ethyl acetate layer showing the highest inhibitory activity in this step were fractionated into 33 small fractions by open silica gel column chromatography. The 33 fractions consisted of CHCl ₃ : EtOH 100: 0, 90:10, 80:20, 70:30, 60:40, 50:50, 40:60, 30:70, 20:80, 10:90 and The fraction obtained by performing each fraction three times for 11 fractions of 0: 100.

Group C of FIG. 3 is deionized water treatment group, group H is H ₂ O ₂ 100 μM treatment group, V is H ₂ O ₂ Vit. For 48 hours before treatment. C 100 μM pre-incubated group, other groups (33 fractions) H ₂ O ₂ Group pre-incubated with sample extract for 48 hours before treatment.

Oxidative stress (%) was induced for a total of 33 small fractions, and five fractions with small induction were selected. Sample 5 of cell viability (%) of FIG. 3 is the second fraction of the 90:10 ratio of CHCl ₃ : EtOH, sample 8 is CHCl ₃ : Second fraction of EtOH = 80: 20, sample 9 is CHCl ₃ : Third fraction of EtOH = 80:20, sample 15 is CHCl ₃ : Third fraction of EtOH = 60:40, sample 16 is CHCl ₃ : The first fraction of EtOH = 50: 50 and the 18th sample were CHCl ₃ : Third fraction of EtOH = 50: 50.

Experimental results of the cell viability (%) of the five fractions selected above are shown in Figure 3, through which the best inhibitory activity fractions of chloroform (chloroform) and ethanol (fraction of the open silica gel column chromatography) ethanol) was found to be the third fraction of 80:20 (v / v) (see FIG. 3).

3. TLC Active substance separation using

CHCl ₃ with good inhibitory activity in fractions of open silica gel column chromatography : EtOH = 80: 20 (v / v) fractions were separated by thin layer chromatography (TLC). Nine bands were identified by TLC and their Rf values were 0.07, 0.19, 0.28, 0.41, 0.54, 0.59, 0.77, 0.92 and 0.96.

The degree of oxidative stress induced by H ₂ O ₂ according to each fraction separated by TLC (see FIG. 4) and cell viability by Aβ (see FIG. 5) were measured. The results confirm the 2 ^nd activity is a good band (R _f value 0.19).

In the case of oxidative stress (%) of FIG. 4, group C is treated with deionized water, and group H is H ₂ O _2. 100 μM treatment group, V is H ₂ O ₂ Vit. For 48 hours before treatment. C 100 μM pre-incubated group, the other groups H ₂ O ₂ Group pre-incubated with sample extract for 48 hours prior to treatment.

In the case of cell viability (%) of FIG. 5, group C was treated with deionized water, group A was treated with Aβ 100 μM, and V was treated with Vit. C 100 μM pre-incubated group, the other groups were pre-incubated with sample extract for 48 hours prior to Aβ treatment. Each number represents an R _f value.

4. HPLC Active substance identification through

The HPLC assay was performed to remove the 2 ^nd group pull band showing the highest activity (R _f value 0.19) active ingredients of part (active compound). The HLPC usage conditions are as follows. Data detection: PDA, 323.3 nm, Column: C _{18 μ} bondapak ^TM (reverse phase column, size: 3.9 x 300 mm), Mobile phase: gradient of 0-100% ethanol in water (total 75 min), Flow rate: 0.5 ml / min, Injection volume: 10 μl (30 mg / ml)

Separation by HPLC confirmed 4-vinyl-2-methoxyphenol having a molecular weight of 105.17 m / z (see Fig. 6).

[ Example 3 ] Confirmation of the Active Effect Using Animals

In To determine the activity of the plantain extract in vivo, a dementia model was established by injecting mice with 410 pmol / horse of Aβ.

Experimental method of alternative behavior (Y-maze test) for measuring memory and learning ability activity and step through latency (manual evasion test) to confirm the change of memory and learning ability are shown in Table 1.

In vivo As an assay, the two trials were run together: mice were divided into four groups, the control group and the negative control group for 4 weeks of normal feed, and the other two groups for 400 and 800 mg / kg per day. In the administration of amyloid beta peptide (Aβ) prior to the experiment, the control group was administered with Aβ (42-1), the rest of the group was administered Aβ (1-42).

In the Y- maze (maze) test for Alternative behavior experiment, the water mixed with the sample to ICR- male mice (male mouse) about three weeks ad Provided as lib (unlimited arbitrary diet). Behavioral experiments were performed in Y-maze with three arms ⓐ, ⓑ and ⓒ by injection of amyloid beta peptide (Aβ) into the ventricle (intracerebroventricular (ICV)). The Y-maze was 32.5 cm long, 15 cm high, and 4 cm wide, with each arm set to A, B, and C, and the arms entered. The mouse was placed in the maze and allowed to move freely for 8 minutes without any stimulation, and the number was calculated by measuring the two hind feet in the arm except for the tail.

Memory and learning step through latency to check for changes in capacity (passive avoidance test, passive avoidance test) tests the food and selection of samples to ICR- male mice as the above conditions, about three weeks ad Provided as lib (unlimited arbitrary diet). Before the test, Aβ was tested for changes in memory and learning ability. In the experiment, the day before entering the experiment, the training was conducted in the same place as the memory box and the behavior experiment was conducted. All mice were subjected to adaptive training (Ming-Ming, No-Shock, Shock) in the test box the day before the experiment, 24 hours later without shock and with light. Experiment for 300 sec per animal.

Group C is a group injected with non-toxic reversed phase Aβ (42-1), group A is injected with Aβ (1-42), PA400 and PA800 plantain (400 mg / kg, 800 mg / kg per day) was ingested with Aβ (1-42), the group was confirmed to improve and maintain memory learning ability significantly in the plantain extract dosing group compared to the Aβ dosing group (see Figure 7).

1 is a diagram showing the damage inhibiting activity of brain neurons by oxidative stress of plantain ethanol extract. (Data is mean ± SD, and it is compared with * P <0.01 C group and # P <0.01 H group.)

2 is a diagram showing the inhibitory activity on brain neuronal cell toxicity for each fraction after fractionation using organic solvents having different polarities in plantain ethanol extract. (Data is mean ± SD, and it is compared with * P <0.01 C group and # P <0.01 H group.)

Figure 3 is a diagram showing the result of the separation of the ethyl acetate fraction separated by solvent partition (solvent partition) by open silica gel column chromatography. (Data is mean ± SD, and it is compared with * P <0.01 C group and # P <0.05 H group.)

Figure 4 is a diagram confirming the degree of oxidative stress induced by H ₂ O ₂ fractions of the active material separated using TLC. (Data is mean ± SD, * P <0.01 compared with C group, # P <0.01 compared with H group, # # P <0.05 compared with H group.)

FIG. 5 is a diagram showing cell viability (%) after adding Aβ to fractions in which an active substance is separated using TLC. Each number represents an R _f value. (Data is mean ± SD, * P <0.05 compared with C group, # P <0.01 compared with A group, # # P <0.05 compared with A group.)

Figure 6 shows the 2 ^nd highest activity HLPC results for identifying the active ingredient of the band.

7 is in Figure showing the results of confirming the protective effect of plantain extract in vivo dementia model. (Data is mean ± SD, * P <0.01 compared with C group, # P <0.01 compared with A group, # # P <0.05 compared with A group.)

Claims (11)

A pharmaceutical composition for the prevention or treatment of degenerative neurological diseases, comprising a plantain ethanol extract or a fraction thereof having 4-vinyl-2-methoxyphenol as an active ingredient. The composition of claim 1, wherein the neurological disease is a disease in which brain nerve cell damage is caused by amyloid beta peptide-induced oxidative stress. The composition of claim 1, wherein the neurological disease is Alzheimer's disease. The composition of claim 1 wherein the fraction is a hexane, chloroform, or ethyl acetate fraction. Pharmaceutical composition for the prevention or treatment of degenerative neurological diseases containing 4-vinyl-2-methoxyphenol. 6. The composition of claim 5, wherein said 4-vinyl-2-methoxyphenol is obtained from plantain extract. The composition of claim 5, wherein the neurological disease is a disease in which brain nerve cell damage is caused by amyloid beta peptide-induced oxidative stress. The composition of claim 5, wherein the neurological disease is Alzheimer's disease. The method of claim 5, wherein 4-vinyl-2-methoxyphenol, Iii) fractionating the plantain extract using an organic solvent having a different polarity to obtain a isolate; Ii) obtaining the brain damage inhibiting active ingredient through the open silica gel column of the isolate obtained in step iii); And Iii) obtaining the active fraction obtained in step ii) by using a reverse phase column HPLC to obtain 4-vinyl-2-methoxyphenol. Degenerative neurological disease prevention or improvement composition for health functional food containing 4-vinyl-2-methoxyphenol. A composition for preventing or improving degenerative neurological disease, comprising a plantain ethanol extract or a fraction thereof having 4-vinyl-2-methoxyphenol as an active ingredient.

Images