KR100951696B1 - Composition for preventing or treating a disease mediated by monoamine oxidase - Google Patents

Abstract

The present invention is Angelica keiskei Koidzumi ) or a composition for the prevention or treatment of diseases mediated by MAO (monoamine oxidase) comprising a compound extracted from the perennial leaf as an active ingredient, and pharmaceuticals and dietary supplement comprising the composition.

Description

Composition for preventing or treating a disease mediated by monoamine oxidase

The present invention relates to a composition for the prevention or treatment of a disease mediated by a monoamine oxidase inhibitor (MAO inhibitor), and more particularly to a MAO inhibitor comprising a plant herb or a compound isolated from the plant herb as an active ingredient. It relates to a composition for preventing or treating a disease mediated by.

 Monoamine oxidase (MAO) is an enzyme that is widely present in mitochondria in animal tissues such as the central nervous system and peripheral tissues, and is responsible for the metabolism of neurotransmitters and hormonal amine compounds. It catalyzes the breakdown of neurotransmitters and hormonal amines derived from food and intestinal bacteria.

RCH ₂ NH ₂ + O ₂ + H ₂ O → RCHO + NH ₃ + H ₂ O ₂

MAO mainly uses catecholamines, indolealkylamines, or derivatives thereof as endogenous substrates, and the oxidation of MAO type A, benzylamine, and phenethylamine, which oxidatively deamines serotonin, norepinephrine, and epinephrine according to substrate specificity, It can be divided into two types, catalyzed MAO B type.

The MAO inhibitors are commonly described in depression (Youdim MB et al., Handbook of Experimental Pharmacology. 90/1, pp. 119-192, 1988; Sambamoorhi U, et al., Med Care. 41 (1), pp180-94, 2003) and migraine (Silberstein SD., Curr Med Res Opin. 17 Suppl 1: s87-93, 2001) and have been used as drugs to delay the progression of Parkinson's disease (Danisi F., Feriatrics. 57 (3)). , pp. 46-50, 2002; Ahlskog JE. Neurology. 60 (3), pp. 381-389, 2003) and other causes of panic symptoms (Sheehan DV., J Clin. Psychiatry., 63 Suppl 14, pp.17-21, 2002), which has been applied as a drug to treat attention deficit and impulsive hyperactivity in young children (Spencer TJ, et al., J Clin Psychiatry. 63 Suppl 12, pp.16-22, 2002 ).

Recently, stimulants of hypersomnia (Annane D, et al., Cochrane Database Syst Rev., (4): CD003218, 2002), silent treatments (Berger I, et al., Isr Med Assoc J. 4 (12), pp. 1135-1137, 2002), smoking cessation aids (Fowler JS, et al., Neurotoxicology. 24 (1), pp. 75-82, 2003; Berlin I, et al., Addction. 97 (10), pp. 1347-1354., 2002; Vessichio JC et al., J. Clin.Psychiatry., 3 (7) .pp.5260-5279, 2002), drug treatments for overcoming social anger disease (Stein DJ, et al., Int. Clin. Psychopharmacol., 17 (4), pp.161-170, 2002).

Among MAO, MAO type B increases with age, and this increase has been reported to be a cause of aging-related gliosis (CJ Fowler et al., J. Neural. Transm. 1980, 49, 1- 20). In addition, MAO-B activity was found to be significantly higher in the brain of Alzheimer's disease patients (P. Dostert et al., Biochem. Pharmacol. 1989, 38, 555-561) and to be highly expressed in astrocytic cells around senile plaques. (Saura et al., Neuroscience 1994, 70, 755-774). In this regard, MAO-B inhibitors may be used to treat dementia and Parkinson's disease.

However, MAO inhibitors inhibit the activity of MAO and at the same time affect the organs affected by sympathetic amines and serotonin, and inhibit the action of other enzymes to interfere with the hepatic metabolism of many drugs. In addition, excessive central stimulation may cause tremor, insomnia, excessive sweating, excitement, hypomania, positional hypotension, dizziness, headache.

In addition, according to D. Bentue-Ferrer et al. (CNS Drugs 1996, 6, 217-236), MAO inhibitors that are not selective for MAO B are at risk of hypertension when ingested dietary tyramine. There is a risk of being induced.

Thus, there is a need for the development of MAO inhibitors that not only improve the side effects but are also selective for MAO B, if possible.

On the other hand, Angelica keiskei Koidzumi is a perennial plant of the Araaceae family and is also known as Sinseoncho, Seonsamcho, and Sinchocho. Myeongilyeop is mainly grown in subtropical regions, and in Korea in 1970s, it is widely used as food such as green juice in Korea. In addition, Myeongilyeop has been used as a folk remedy for the treatment of hypertension, liver disease, neuralgia, etc., and physiological activities such as antihyperlipidemia, lowering blood pressure, anticancer action, gastric acid secretion are reported.

There is an active research on the active compounds present in the leaf. It is known that there are chalcone compounds such as xantho-angerol having a structure of Formula 1 and 4-hydroxydericin having a structure of Formula 2, and these compounds have anticancer effects (Yoshiyuki Kimura et al., Antitumor). and antimetastatic activities of 4-hydroxyderricin isolated from Angelica keiskei roots, Planta Med, 2004, 70, 211-219), Inamori Yoshihiko et al., Antibeacterial activity of two chalcones, xanthoangelol and 4-hydroxyderricin, isolated from the root of Angelica keiskei KOIDZUMI, Chemical & Pharmaceutical bulletin, Vol 39, No. 6, pp. 1604-1605), hypoglycemic effect (Tatsuji Enoki et al., Antidiabetic activities of chalcones isolated from a Japanese Herb, Angelica keiskei, J. Agric Food Chem., 55 (15), 6013-6017), has been reported to have a hepatoprotective effect (Korean Patent Publication 2006-106065).

However, until now, the extract of Mt. Ilyun and compounds isolated therefrom have not been reported to have a MAO inhibitory effect, and there have been no reports of an improvement effect of various indications by the MAO inhibitory effect.

Accordingly, the present inventors have completed the present invention as a result of trying to find a plant herbal medicine having low toxicity or side effects even if taken for a long time while having MAO inhibitory activity.

Accordingly, it is an object of the present invention to provide a composition derived from a plant herbal medicine having MAO inhibitory activity.

Another object of the present invention is to provide a composition comprising a compound derived from a plant herb having MAO inhibitory activity.

Still another object of the present invention is to provide a medicine comprising the composition.

Another object of the present invention to provide a dietary supplement comprising the composition.

In order to achieve the above object, the present invention provides a composition for the prophylaxis or treatment of diseases mediated by MAO comprising Myeongil lobe as an active ingredient.

The present invention also provides a composition for the prophylaxis or treatment of a disease mediated by MAO comprising the compound of formula 3 or a pharmaceutically acceptable salt thereof as an active ingredient:

In Chemical Formula 3,

R ¹ is a hydroxy group or C ₁ -C ₄ alkoxy,

또는

이다. R ² is

or

to be.

The present invention also provides a medicine comprising the composition.

In addition, the present invention provides a health supplement comprising the composition.

Hereinafter, the present invention will be described in more detail.

The present inventors have conducted studies on various plant herbals to find plant herbals having MAO inhibitory activity. As a result, they have found that Mt.

Therefore, in one aspect, the present invention provides a composition for the prevention or treatment of diseases mediated by MAO, which comprises the unicolored leaf as an active ingredient.

Myeongil leaf, which is an active ingredient of the composition, may be included in the form of an extract, or may be included as a pulverized product of the herbal medicine or a dry pulverized product of the herbal medicine.

The extract of Myeongil Leaf may be a crude extract of water, a C ₁ -C ₄ alcohol, and a solvent selected from the group consisting of a combination thereof. The crude extract of the day leaf is preferably a C ₁ -C ₄ alcohol extract, more preferably an extract of ethanol, more preferably 70 to 100% ethanol extract. When extracting with a solvent, the extraction may be heat extraction, cold extraction, reflux extraction, or ultrasonic extraction. This extraction may be carried out at room temperature, but for more efficient extraction, it may be carried out under warm conditions, preferably at a temperature of 35 to 100 ° C, more preferably 40 to 80 ° C. Extraction time may be performed for 1 to 4 days, preferably 3 to 12 hours, and may vary depending on conditions such as an extraction solvent and an extraction temperature. The extraction may be extracted one or more times several times in order to obtain a larger amount of the active ingredient, preferably one to five times, more preferably three times the continuous extraction can be used combined extract.

The composition comprising Myeongil Lobe according to the present invention may include Myeongil Lobe as a crude extract as described above, but may also comprise the crude extract as a soluble extract of a nonpolar solvent obtained by further extracting with a nonpolar organic solvent. Hexane, dichloromethane, ethyl acetate, butanol, and the like may be used as the nonpolar organic solvent, but is not limited thereto. Preferably, the crude ethanol crude extract is extracted with hexane to obtain a hexane fraction, and the remaining fractions are suspended in water, followed by dichloromethane, ethyl acetate and n-butanol in order to obtain soluble fractions of each solvent. In addition, the above extract can be obtained as a soluble extract of a nonpolar solvent. Among the soluble extracts of the nonpolar solvent, dichloromethane soluble fraction and ethyl acetate soluble fraction are preferred because of their high MAO inhibitory activity.

The extract extracted by the method as described above may be used as it is, but may be used in the form of concentrated X, or may be concentrated and then lyophilized to be used as a lyophilized form. Preferably, it is used in concentrated form so that it can be used at a sufficient concentration.

The soluble fraction of the nonpolar solvent may be further purified by chromatography to yield the active compound of the sunleaf. The dichloromethane soluble fraction of the soluble fraction of the non-polar solvent can be purified by chromatography to separate xanthoangele and 4-hydroxydericin, which are known as pharmacologically active ingredients of the daylight. The present inventors confirmed that the compounds of xanthoangelol and 4-hydroxyderycin isolated also have MAO inhibitory activity.

Accordingly, in another aspect of the present invention, the present invention provides a composition for the prevention or treatment of diseases mediated by MAO, comprising as an active ingredient a compound of formula 3 or a pharmaceutically acceptable salt thereof:

(3)

In Chemical Formula 3,

R ¹ is a hydroxy group or C ₁ -C ₄ alkoxy,

또는

이다. R ² is

or

to be.

As described above, the compound of Formula 3 may be obtained from the extract of Sunil Leaf by a method known to be capable of separating the substance, or obtained by chemical synthesis using conventional techniques known in the art. You may lose.

The compound of formula 3 is preferably xanthoangelol or hydroxydericin.

According to an embodiment of the present invention, the compound of formula 3 may be obtained from the dichloromethane soluble fraction. The dichloromethane soluble fraction was subjected to silica gel column chromatography to elute with hexane / dichloromethane (1: 3) to dichloromethane / methanol (30: 1) to obtain four small fractions (Fr. I -IV). Among them, Fr.I can be subjected to Lichroprep RP-18 column chromatography, eluting with 80% methanol to obtain xanthoangelol as a yellow needle, and Fr.II can be obtained by Lichroprep RP-18 column chromatography. It can be carried out eluting with 80% methanol to obtain 4-hydroxydericin as a yellow powder.

The composition of the present invention comprising the compound of formula (I) or the compound of Formula 3 as an active ingredient can be used for the prevention or treatment of diseases mediated by MAO. The disease mediated by the MAO refers to any disease in which MAO inhibitory activity may be beneficial for the treatment or prevention of the disease. Such diseases include, but are not limited to, depression, migraine, Parkinson's disease, Alzheimer's disease, neuroglia, panic disorder, attention deficit disorder, excess sleep, or withdrawal symptoms caused by alcohol or nicotine.

It is demonstrated in the examples below that the compositions according to the invention have MAO inhibitory activity. Specifically, the ethanol extract of Myeongil Leaf extracted according to one embodiment of the present invention, the ethanol extract of each soluble fraction extracted in the order of hexane, dichloromethane, ethyl acetate, butanol, and separated from the dichloromethane soluble fraction The inhibitory activity of MAO enzymes obtained from rat brain or liver against santoangerol or hydroxydericin was found to be excellent.

The composition for the prevention or treatment of diseases mediated by MAO having active compound of the present invention or the compound of formula 3 according to the present invention can be used as a raw material of medicines effective for the treatment or prevention of diseases mediated by MAO. have.

Accordingly, the present invention in another aspect, the present invention provides a pharmaceutical composition comprising the composition or the pharmaceutically acceptable carrier or additive containing the compound of formula (3).

The medicament may be formulated in conventional pharmaceutical formulations known in the art. The medicament may be formulated and administered in any dosage form including, but not limited to, oral, injectable, suppository, transdermal, and non-administrative agents, but preferably liquids, suspensions, powders, granules, It may be formulated into a formulation for oral administration such as tablets, capsules, pills, or excipients.

When formulated into each of the above formulations, it may be prepared by the addition of a pharmaceutically acceptable carrier or additive necessary for the preparation of each formulation. Representatively, when formulated into a dosage form for oral administration, one or more of a diluent, a lubricant, a binder, a disintegrant, a sweetener, a stabilizer, and a preservative may be used as the carrier, and as an additive, flavors, vitamins, and antioxidants may be used. One or more types can be selected and used out of them.

The carrier and the additive may be any pharmaceutically acceptable, and specifically, lactose, corn starch, soybean oil, microcrystalline cellulose, or mannitol as a diluent, magnesium stearate or talc as a lubricant, and polyvinylpyrrolidone as a binder. Or hydroxypropyl cellulose is preferred. In addition, as a disintegrant, carboxymethyl cellulose calcium, sodium starch glycolate, potassium polyacrylic acid, or crospovidone, sweetener as white sugar, fructose, sorbitol, or aspartame, stabilizer as carboxymethyl cellulose sodium, beta-cyclodextrin, As lead, or xanthan gum, and preservative, methyl paraoxybenzoate, propyl paraoxybenzoate, or potassium sorbate is preferable.

In addition to the above ingredients, in order to enhance the taste as a known additive, natural flavors such as plum flavor, lemon flavor, pineapple flavor, herbal flavor, natural pigments such as natural fruit juice, chlorophyllin, flavonoid, fructose, honey, sugar alcohol, sugar Sweetening ingredients such as, or may be used by mixing an acidulant such as citric acid, sodium citrate.

The drug is effective in preventing or treating a disease in which MAO inhibitors are effective, including depression, migraine, Parkinson's disease, Alzheimer's disease, neuroglia, panic disorder, attention deficit disorder, hyperactivity, or withdrawal symptoms caused by alcohol or nicotine. For this purpose, the total daily dose of the active ingredient may be 0.01 to 0.5 g / kg as an extract or a compound, and 0.1 to 5.0 g / kg as a crude drug powder. The dosage may be appropriately increased or decreased depending on the route of administration, disease progression, sex, age, weight, and the like.

In addition, the composition for the prevention or treatment of diseases mediated by MAO having the active ingredient of the compound or formula 3 of the present invention according to the present invention of the health supplement foods effective in the prevention or treatment of diseases mediated by the MAO It can be used as raw material.

Therefore, in yet another aspect of the present invention, the present invention provides a dietary supplement comprising the composition of the light or the compound of Formula 3, and a food or acceptable carrier or additive.

Such dietary supplements may be formulated in the form of conventional dietary supplements known in the art. The dietary supplement may be prepared, for example, as a powder, granules, tablets, capsules, suspensions, emulsions, syrups, liquids, extracts, teas, jelly, extracts, or beverages. As the food acceptable carrier or additive, any carrier or additive known to be usable in the art may be used to prepare a formulation to be prepared. As such, the health supplement of the present invention processed in various forms can be used not only for the prevention of various diseases mediated by MAO, but also easy to take and useful.

In addition, Myeongil leaf is currently being widely used as a food, Myeongil leaf extract or a compound of formula 3 derived from it can be seen that there is no problem of toxicity or side effects, the pharmaceutical and health supplement according to the present invention is to prevent or There is an advantage that can be used with confidence even for long time use for therapeutic purposes.

As described above, the composition comprising the compound of the present invention or the compound of formula 3 as an active ingredient according to the present invention can be used for the prevention or treatment of various diseases mediated by MAO due to its excellent MAO inhibitory activity, and proved safety By using the raw herbal medicines as raw materials, there is an advantage that can solve various side effects that can be exhibited by the conventional MAO inhibitor.

Hereinafter, the present invention will be described in more detail with reference to the following examples. However, these examples are only for the understanding of the present invention, and the scope of the present invention is not limited by them in any sense.

Example 1 Bright Day Material Preparation

Myeongilyeop was purchased from Myeongilyeop Farm in Wonju, Gangwon-do in July 2006, dried at 80 ℃ and pulverized to powder.

Example 2: Preparation of Sunil Leaf Extract

10 kg of the fresh-leaf powder prepared in Example 1 was added to 100 L of 70% ethanol and refluxed at 80 ° C. for 4 hours, and then concentrated under reduced pressure to obtain a concentrate of 2.8 kg.

Example 3 Preparation of Nonpolar Solvent Fractions

1000 g of the 70% ethanol concentrate prepared in Example 2 was suspended in 90% methanol, extracted with hexane, and concentrated under reduced pressure to obtain a hexane fraction (110 g). Then, the 90% methanol layer was concentrated, suspended with water, extracted with dichloromethane, and concentrated under reduced pressure to obtain a dichloromethane fraction (56 g). Then extracted with dichloromethane and the remaining water layer was extracted with ethyl acetate and concentrated under reduced pressure to obtain an ethyl acetate fraction (45g), extracted with ethyl acetate and extracted with butanol and concentrated under reduced pressure to give a butanol fraction (138g). . Finally, the remaining water layer was concentrated under reduced pressure to obtain a water fraction (400 g).

Example 4: Isolation of Compounds

The dichloromethane fraction (56 g) obtained in Example 3 was subjected to silica gel column chromatography, eluting with hexane / dichloromethane (1: 3) to dichloromethane / methanol (30: 1) to obtain four small fractions (Fr. I). -IV) was obtained. Among them, Fr. I (15 g) was subjected to Lichroprep RP-18 column chromatography to elute with 80% methanol to afford yellow needles Compound 1 (3.1 g) on methanol and dichloromethane mixed solution.

Next, the Fr. II (6 g) was subjected to Lichroprep RP-18 column chromatography to concentrate the solution eluted with 80% methanol, and concentrated to give Compound 2 (450 mg) as a yellow powder.

Separately, the butanol fraction (138 g) obtained in Example 3 was subjected to silica gel column chromatography, eluting with dichloromethane / methanol (10: 1) to dichloromethane / methanol (3: 1) to give four small fractions (Fr. I-IV). Among them, Fr. II (13 g) was subjected to silica gel column chromatography, eluted with dichloromethane / methanol 5: 1, and recrystallized in methanol solution to obtain compound 3 (300 mg) as yellow needles.

Compounds 1 to 3 were analyzed by NMR and MS, and the results are shown below.

(1) Compound 1: xanthoangelol

Appearance: Yellow couch.

EI-MS: m / z 392 [M] ⁺ (C ₂₅ H ₂₈ O ₄ ),

¹ H-NMR (600 MHz, CDCl ₃ ): δ 1.59 (3H, s, 7′-CH ₃ ), 1.63 (3H, s, 8′-CH ₃ ), 1.85 (3H, s, 3′-CH ₃ ) , 2.08 (2H, m, 4 μs-H), 2.10 (2H, m, 5 μs-H), 3.49 (1H, d, J = 7.1 Hz, 1 μs-H), 5.05 (1H, m, 6 μs) -H), 5.30 (1H, t, J = 7.1 Hz, 2 Hz-H), 6.43 (1H, d, J = 8.9 Hz, 5'-H), 6.88 (2H, d, J = 8.5 Hz, 3 , 5-H), 7.46 (1H, d, J = 15.4 Hz, α), 7.55 (2H, d, J = 8.5 Hz, 2,6-H), 7.72 (1H, d, J = 8.9 Hz, 6 '-H), 7.83 (1H, d, J = 15.4 Hz, b), 13.88 (1H, s, 2'-OH).

¹³ C-NMR (150 MHz, CDCl ₃ ): δ 16.5 ( ₃ ˝- C H ₃ ), 17.9 (7 ˝- C H ₃ ), 21.9 (1 ˝-C), 25.9 (8 ˝-C), 26.6 ( 5 ˝-C), 39.9 (4 ˝-C), 108.1 (5 ', 3 ˝-C), 114.2 (1'-C), 114.3 (3'-C), 116.2 (3, 5-C), 118.2 (aC), 121.2 (2˝-C), 123.9 (6˝-C), 127.9 (1-C), 130.8 (2, 6'-C), 139.9 (7˝-C), 144.4 (bC) , 162.1 (4, 2'-C), 164.1 (4'-C), 192.5 (C = O)

(2) Compound 2: hydroxyderricin

Appearance: Yellow powder.

EI-MS: m / z 338 [M] ⁺ (C ₂₁ H ₂₂ O ₄ ),

¹ H-NMR (600 MHz, CDCl ₃ ): δ 1.60 (3H, s, 4′-CH ₃ ), 1.71 (3H, s, 5′-CH ₃ ), 3.31 (1H, d, J = 7.0 Hz, 1 ˝-H), 3.82 (3H, s, 4-OCH ₃ ), 5.30 (1H, t, J = 7.0 Hz, 2 ˝-H), 6.41 (1H, d, J = 9.0 Hz, 5'-H) , 6.79 (2H, d, J = 8.5 Hz, 3,5-H), 7.36 (1H, d, J = 15.4 Hz, α), 7.70 (2H, d, J = 9.4 Hz, 2,6-H) 7.70 (1H, d, J = 9.4 Hz, 6'-H), 7.72 (1H, d, J = 15.4 Hz, b), 13.38 (1H, s, 2'-OH).

¹³ C-NMR (150 MHz, CDCl ₃ ): δ 18.0 (3′-CH ₃ ), 21.9 (1′-C), 26.0 (4′-C), 56.0 (4-OCH ₃ ), 102.4 (5'- C), 108.1 (5 ', 3'-C), 114.8 (1'-C), 114.3 (3-C), 117.8 (5-C), 118.1 (3', aC), 122.2 (2'-C) ), 127.7 (1-C), 129.4 (6'-C), 130.8 (6-C), 132.2 (3˝-C), 144.4 (bC), 163.2 (4, 2'-C), 163.5 (4 '-C), 158.6 (4-C), 163.5 (4'-C), 192.7 (C = O)

(3) Compound 3: Cynaroside

Appearance: Yellow couch.

¹ H-NMR (600 MHz, DMSO d ₆ + D ₂ O) δ 5.04 (1H, d, J = 7.5 Hz, G1), 6.44 (H, d, J = 2.0 Hz, 6-H), 6.73 (1H, s, 3-H), 6.80 (1H, d, J = 2.1 Hz, 8-H), 6.91 (1H, d, J = 8.4 Hz, 5'-H), 6.91 (1H, d, J = 8.4 Hz , 5'-H), 7.40 (1H, d, J = 2.2 Hz, 2'-H), 7.44 (1H, d, J = 8.4 Hz, 6'-H).

¹³ C-NMR (150 MHz, DMSO d ₆ + D ₂ O): δ 60.8 (G6-C), 69.8 (G4-C), 73.3 (G2-C), 76.4 (G3-C), 77.3 (G5-C ), 95.3 (C-8), 99.9 (6-C), 100.2 (G1-C), 105.7 (10-C), 113.7 (2'-C), 116.2 (3-C), 116.3 (5'- C), 119.6 (6'-C), 121.7 (1'-C), 145.9 (3'-C), 150.1 (4'-C), 151.1 (5-C), 157.3 (9-C), 163.2 (7-C), 164.8 (2-C), 182.2 (4-C).

Experimental Example 1: Reagents and Experimental Preparation

1-1. reagent

Serotonin, benzylamine, ion exchange resin Amberlite CG-50, etc. used for the measurement of the MAO enzyme activity were manufactured by Sigma, and other reagents for column chromatography and sample extraction were used with a special reagent (Merck).

1-2. Experimental animal

Sprague Dawley male rats, 5 weeks old, were supplied by Orient Bio Co., Ltd. and were supplied with general solid feed and water freely in the animal breeding room with temperature control 23 ± 3 ℃, humidity 50 ± 10%, and lighting for 12 hours 1 Daytime adaptation was used for the experiments.

Experimental Example 2: Brain MAO-A Inhibitory Activity Test

2-1. Enzyme source preparation

Rats were anesthetized in an ether-doped anesthesia, opened, and then bled by blood collection from the left ventricle, and immediately dissected the skull to extract the brain. The extracted brain was washed with 0.01 M phosphate buffered solution (PBS; pH 7), 9 mL of cold 0.25 M sucrose solution per 1 g of wet weight was added, and homogenized with a Turrax homogenizer for 1 minute.

The homogenate was centrifuged at 700 x g for 20 minutes at 4 ° C, the supernatant was taken and again centrifuged at 18,000 x g for 20 minutes at high speed, and the supernatant was discarded and the pellet was suspended and suspended in 5 mL of PBS per 1 g weight. It was used as an enzyme source.

2-2. Enzyme Activity Measurement

0.5 mL of the enzyme source prepared above was placed in a test tube, and 0.5 mL of 0.1 mM serotonin solution was added as a substrate solution, and the resultant was incubated for 90 minutes in a 37.5 ° C incubator. The reaction was stopped by heating for 3 minutes in a 95 ° C. water bath, immediately centrifuged at 700 × g, and 1.0 mL of the supernatant was poured into a previously prepared Amberlite GC-50 (H ⁺ form) column (0.6 × 4 cm). After sufficiently washing the resin with distilled water (40 mL or more), 3 mL of 4 N acetic acid solution was poured into the resin, and the eluate was received in a test tube and the absorbance was measured at 277 nm.

Separately, a correction group with a substrate solution at the reaction end point instead of the reaction start point was carried out with the test group. The test was carried out in the 70% ethanol concentrate prepared in Example 2, the hexane fraction obtained in Example 3, the dichloromethane fraction, the butanol fraction, the aqueous layer fraction, and the xantho-angerol obtained in Example 4, 4-hydroxydericin , And sinarosides. Each concentrate or fraction was dissolved in distilled water to a 1 mg / mL solution, and 1 (1 mg / mL) and 1/10 (0.1 mg / mL) dilutions were used as the sample solution. The three compounds obtained in Example 4 were diluted to a concentration of 3-50 μg / mL to measure the inhibitory activity of MAO-A.

Based on the control group of each test group, the change in enzyme activity at the time of sample administration according to the change in concentration was calculated.

The experimental results are shown in Table 1 below.

TABLE 1

According to the results shown in Table 1, all of the ethanol extract, dichloromethane extract, ethyl acetate extract, and butanol extract of Myeongil Leaf were shown to inhibit MAO A activity. The ethanol extract of Sunil leaf was shown to significantly inhibit the enzymatic activity of MAO-A in vitro, and the IC ₅₀ value for MAO-A of the ethanol extract was 0.51 mg / mL. The ethyl acetate fraction in the solvent fraction was found to have the strongest inhibitory activity against MAO-A, and the IC ₅₀ value for the MAO-A of the ethyl acetate fraction was 0.085 mg / mL. The dichloromethane fraction also showed inhibitory activity against MAO-A with an IC ₅₀ value of 0.30 mg / mL. Xantho angerol of the isolated compound showed an inhibitory activity with an IC ₅₀ value of 0.05 mM for MAO-A, and no inhibitory activity was observed for 4-hydroxydericin and sinaroside.

Experimental Example 3: Determination of Liver MAO-B Activity

3-1. Enzyme source preparation

Rats were anesthetized in an ether-doped anesthesia bottle, opened, and blood was collected from the left ventricle and blood was immediately extracted. The extracted liver was washed with 0.01 M phosphate buffered saline (PBS; pH 7), 9 mL of cold 0.25 M sucrose solution of 9 mL per 1 g of wet weight was added, and homogenized with a homogenizer for 1 minute.

The homogenate was centrifuged at 700 × g for 20 minutes at 4 ° C., the supernatant was taken and again centrifuged at 18,000 × g for 20 minutes at high speed. The supernatant was discarded and the pellet was submerged in 5 mL of PBS / g. Suspension was used as an enzyme source.

3-2. Enzyme Activity Measurement

Enzyme activity was measured according to the method of McWeen et al. (McWeen, C.M. et al., J. Lab. Clin. Med., 62, pp. 766-776, 1963).

0.5 mL of the enzyme source was placed in a test tube, and 0.5 mL of a 4 mM benzylamine-HCl solution was added as a substrate solution, and incubated for 90 minutes in a 37.5 ° C thermostat. In order to stop the reaction, 0.2 mL of 60% perchloric acid solution was added, and 4 mL of cyclohexane was added thereto, followed by shaking. Then, centrifugation was performed at 700 x g for 20 minutes, and a cyclohexane layer was taken to measure absorbance at 242 nm.

Separately, a correction group with a substrate solution at the reaction end point instead of the reaction start point was carried out with the test group. The test was carried out in the 70% ethanol concentrate prepared in Example 2, the hexane fraction obtained in Example 3, the dichloromethane fraction, the butanol fraction, the aqueous layer fraction, and the xantho-angerol obtained in Example 4, 4-hydroxydericin , And sinarosides. Each concentrate or fraction was dissolved in distilled water to a 1 mg / mL solution, and 1 (1 mg / mL) and 1/10 (0.1 mg / mL) dilutions were used as the sample solution. The three compounds obtained in Example 4 were diluted to a concentration of 3-50 μg / mL to measure the inhibitory activity of MAO-B.

The change in concentration and the change in enzyme activity according to the administration of the sample were calculated based on the control group of each test group.

The experimental results are shown in Table 2 below.

TABLE 2

According to the results shown in Table 2, the extract of Myeongil-Yeop showed an overall stronger inhibitory activity against MAO-B than the inhibitory activity against MAO-A. The IC ₅₀ value for MAO-B of the ethanol extract was 0.33 mg / mL. The strongest inhibitory activity against MAO-B was identified in the dichloromethane fraction, and the IC ₅₀ value for MAO-B in the dichloromethane fraction was 0.055 mg / mL. The IC ₅₀ values for the MAO-B of the ethyl acetate fraction and the butanol fraction were measured at 0.13 mg / mL and 0.85 mg / mL, respectively. The IC ₅₀ values for the MAO-B of the hexane and water fractions were 1.9 mg / mL, respectively. And 2.75 mg / mL showed weak inhibitory activity. Among the isolated compounds, xantho angerol also showed strong inhibitory activity against MAO-B, IC ₅₀ value was 0.04 mM, and 4-hydroxydericin showed very strong activity with IC ₅₀ value of 3.85 μM. It became. On the other hand, cinnaroside did not show inhibitory activity in MAO-B as in MAO-A.

In conclusion, the extract of M. ileum inhibits both MAO-A and MAO-B throughout the soluble fraction of ethanol extract and non-polar solvent. It can be seen that it can be used for treatment and prevention. In addition, it can be seen that xantho-angerol and 4-hydroxyderincin, which are compounds extracted from the day, also exhibit MAO inhibitory activity and can be used for the treatment and prevention of diseases mediated by MAO. In addition, the extract of Myeongil Leaf and the compound released therefrom have a stronger inhibitory activity on MAO-B than MAO-A, which may reduce the risk of hypertension induced when ingesting dietary tyramine. .

Claims (10)

Pharmaceutical composition for the prevention or treatment of depression comprising Angelica keiskei Koidzumi as an active ingredient. A composition for health supplement food for preventing or improving depression comprising Myeong Il-Yop ( Angelica keiskei Koidzumi ) as an active ingredient. The crude extract according to claim 1 or 2, wherein the light leaf is contained as a crude extract of a crude product itself, a dry ground product of the herbal medicine, or a solvent selected from the group consisting of water, C ₁ -C ₄ alcohols, and combinations thereof. A composition, characterized in that. The composition according to claim 1 or 2, wherein the light leaf is contained as an extract of a solvent selected from the group consisting of hexane, dichloromethane, ethyl acetate, and combinations thereof. A pharmaceutical composition for preventing or treating depression comprising the compound of formula 3 or a pharmaceutically acceptable salt thereof as an active ingredient: (3)

In Chemical Formula 3, R ¹ is a hydroxy group or C ₁ -C ₄ alkoxy, R ² is

or

to be. A composition for preventing or ameliorating depression comprising the compound of Formula 3 or a pharmaceutically acceptable salt thereof as an active ingredient: (3)

In Chemical Formula 3, R ¹ is a hydroxy group or C ₁ -C ₄ alkoxy, R ² is

or

to be. The composition of claim 5 or 6, wherein the compound of Formula 3 is xanthoangelol or hydroxydericin. The composition according to claim 1 or 5, which is in the form of a powder, granule, tablet, capsule, suspension, emulsion, syrup, liquid, aerosol, extract, injectable, transdermal, or suppository. delete The composition according to claim 2 or 6, which is in the form of a powder, granule, tablet, capsule, suspension, emulsion, syrup, liquid, aerosol, extract, tea, jelly, or beverage.