KR101854762B1 - Composition for preventing or treating of depression and anxiety comprising alternariol monomethyl ether - Google Patents

Abstract

The present invention relates to a composition for preventing or treating depression and anxiety, comprising alternariol monomethyl ether (AME) as an active ingredient. The AME of the present invention is capable of selectively, reversibly, and competitively inhibiting activities of monoamine oxidase A (MAO-A) associated with the onset of depression and anxiety. The composition shows little cytotoxicity, and thus can be safely used in pharmaceutical or food compositions to prevent or treat depression and anxiety.

Description

TECHNICAL FIELD The present invention relates to a composition for preventing or treating depression and anxiety, which comprises, as an active ingredient,

The present invention relates to a composition for preventing or treating depression and anxiety, which comprises alternaria monomethyl ether (AME) as an active ingredient. More particularly, the present invention relates to a composition for preventing or treating depression And pharmaceutical or food compositions for the prevention or treatment of anxiety.

Korea has undergone rapid development over the last fifty years in such a way that it is hard to find any analogy in the world as it has experienced the industrialization, urbanization, westernization, nuclear family, advanced education, aging and informationization process in a short period of time. These rapid developments have led to high economic growth, but they have been associated with aging, excessive disparities in wealth, family dismemberment, alienation, neurological diseases such as Parkinson's disease due to psychological stress from intense competition, dementia, , Depression, and anxiety disorder (neuropsychiatric disorder) are increasing.

The onset of this neuropsychiatric disorder plays an important role in the metabolism of monoamines in vivo, and monoamine oxidase (MAO, E.C. 1.4.3.4) is known to be involved as an enzyme.

Specifically, monoamine oxidase (MAO) is an enzyme that is widely distributed in the central nervous system and peripheral tissues. It catalyzes the oxidative deamination of amine compounds and is derived from monoamine compounds and intestinal bacteria Which break down hormonal amines. That is, the neurotransmitter is decomposed by the monoamine oxidase, resulting in the deficiency of the monoamine compound in vivo, resulting in the neuropsychiatric disorder.

This monoamine oxidase (MAO) is mainly used as an endogenous substrate such as dopamine, tyramine, epinephrine or norepinephrine, and serotonin is selectively eliminated by substrate specificity The amino acid can be divided into two types: A-type (type A, MAO-A), B-type (type B, MAO-B) that selectively deaminates phenylethylamine and benzylamine (Youdim MB et al. , 2006, Nat. Rev. Neurosci., 7: 295-309). Among these, MAO-A is associated with the onset of neuropsychiatric disorders such as depression and anxiety, and MAO-B is associated with the onset of neurological diseases such as Alzheimer's diseases and Parkinson's diseases.

Thus, MAO inhibitors have been used to prevent or treat neuropsychiatric disorders domestically and internationally, and the MAO inhibitors include MAO-A selective inhibitors, MAO-B selective inhibitors, and MAO-A / And irreversible inhibitors [Malcomson T et al. , 2015, FEBS J., 282: 3190-3198; Mostert S et al. , 2015, Chem. Med. Chem., 10: 862-873).

(1990, Br. J. Clin. Pharmacol., 30: 805-816), Fowler et al. (1990), and the like in connection with MAO-A selective inhibitors that can prevent or treat depression and anxiety, Neuropsychopharmacology, 35: 623-631), Gentili F et al (2006, J. Med. Chem., 49: 5578-5586), and Lotufo-Neto F et al. (1999, Neuropsychopharmacology, 20: 226-247 ) Reports that moclobemide, brofaromine, toloxatone, amifuraline, and CX157 can be used as reversible inhibitors of MAO-A. (1997, Arch. Biochem. Biophys., 337: 137-142) and Mattsson C et al. (2014, Eur J. Med. Chem., 73: 177-186) have discovered novel beta-carbolines derivatives and coumarin derivatives, which have been reported to be available as selective inhibitors of MAO-A . However, the MAO-A reversible and selective inhibitor is a chemically synthesized substance that causes hepatotoxicity and somatic hypotension, and has the problem that side effects such as insomnia, excitement, and convulsions occur when overdose.

Therefore, there is a demand for the development of medicines or health functional foods which are safe for long-term use and can treat or ameliorate depression and anxiety.

On the other hand, the ether or rheol monomethyl ether (AME) is a mycotoxin produced mainly in the strain of the genus Alternaria, especially in the genus Alternaria brassicae , and has a toxic effect in animals.

However, it has not been reported that the ether or rheol monomethyl ether (AME) isolated from Alteraria brassicaceae has the effect of preventing or treating depression and anxiety.

Accordingly, the inventors of the present invention have found that, while searching for a substance that is safe for long-term use and can prevent or treat depression and anxiety, it has been found that the use of ether or ranol monomethyl ether (AME), isolated from Alternaria brassicae , It can be used as a prophylactic or therapeutic agent for mental disorders, and the present invention has been completed.

Accordingly, a technical problem to be solved by the present invention is to provide a composition for preventing or treating depression and anxiety.

In order to solve the above technical problems, the present invention provides a pharmaceutical composition for preventing or treating depression and anxiety comprising, as an active ingredient, alternaria monomethyl ether or a pharmaceutically acceptable salt thereof .

In addition, the present invention provides a food composition for preventing or ameliorating depression and anxiety, which comprises, as an active ingredient, an ether or a rhein monomethyl ether or a pharmaceutically acceptable salt thereof.

The term "depression" as used in the present invention is a feeling of feeling and feeling of sadness, tastelessness, loss of emotion, lack of pleasure (lack of pleasure), grief, agitation or delay, guilt and worthlessness, , ≪ / RTI > and delusions.

As used herein, the term "anxiety" refers to a psychological disorder in which the physical or behavioral symptoms such as headache, increased heart rate, increased respiratory rate, abnormal gastrointestinal symptoms, panic disorder, sweating, It means disorder.

The term "prevention ", as used herein, refers to inhibiting the occurrence of a disease or disease in an individual who has never been diagnosed as having a disease or disease, but has a tendency to become susceptible to such disease or disease.

The term "treatment ", as used herein, refers to (a) inhibiting the development of a disease or disorder, (b) relieving the disease or disorder and (c) eliminating the disease or disorder.

The term "individual" as used herein means a mammal such as a monkey, a cow, a horse, a pig, a sheep, a dog, a cat, a rat, a mouse, a chimpanzee, etc. including a human having a disease whose symptoms may be improved by administering the composition of the present invention Of mammals.

Alternaria monomethyl ether (AME), an active ingredient of the composition of the present invention, is a compound having a structure represented by the following formula (1).

The ether or ranol monomethyl ether may be isolated from the genus Alteraria or chemically synthesized and preferably isolated from the strain of Alternaria brassicae .

The ether or ranol monomethyl ether of the present invention selectively, reversibly and competitively inhibits the activity of monoamine oxidase A (MAO-A), which is associated with the onset of depression and anxiety.

Therefore, the composition containing the ether or ranol monomethyl ether of the present invention as an active ingredient can be usefully used for preventing or treating depression and anxiety.

The ether or ranol monomethyl ether contained as an active ingredient in the composition of the present invention is contained in an amount of 0.001 to 50% by weight, preferably 0.01 to 30% by weight, more preferably 0.01 to 10% by weight, based on the total weight of the total composition ≪ / RTI > When the content of the ether or ranol monomethyl ether is less than 0.001% by weight, the MAO-A inhibitory activity is insufficient. When the content of the ether or ranol monomethyl ether is more than 50% by weight, the effect of increasing the content is not proportional, There is a problem in that it can not be secured.

As one specific use of the present invention, the composition of the present invention can be used as a pharmaceutical composition for preventing or treating depression and anxiety.

When the composition of the present invention is used as a pharmaceutical composition, a pharmaceutically acceptable carrier other than the ether or the ranol monomethyl ether may be further contained as an active ingredient. The pharmaceutically acceptable carriers to be contained in the pharmaceutical composition of the present invention are those conventionally used in the present invention and include carbohydrate-type compounds (e.g., lactose, amylose, dextrose, sucrose, sorbitol, mannitol, starch, Corn oil, cottonseed oil, cottonseed oil, cottonseed oil, cottonseed oil, cottonseed oil, cottonseed oil, cottonseed oil, cottonseed oil, cottonseed oil, cottonseed oil, But are not limited to, polyethylene glycol, methylcellulose, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil, and the like. The pharmaceutical composition of the present invention may further contain a lubricant, a wetting agent, a sweetening agent, a flavoring agent, an emulsifying agent, a suspending agent, a preservative, etc., in addition to the above components. Suitable pharmaceutically acceptable carriers and formulations are described in detail in Remington ' s Pharmaceutical Sciences (19th ed., 1995).

The pharmaceutical composition may be formulated in the form of powders, granules, tablets, capsules, suspensions, emulsions, oral formulations such as aerosols, external preparations, suppositories, and sterilized injection solutions according to a conventional method.

The pharmaceutical composition may prevent or treat depression and anxiety. The method of treatment includes administering the pharmaceutical composition in a pharmaceutically effective amount by various routes in a mammal such as a mouse, a mouse, a domestic animal, or a human. The administration method may be carried out in any manner, for example, by oral, rectal or intravenous infusion, intraperitoneal administration, intramuscular administration, or intracerebral administration.

The dosage of the pharmaceutical composition may vary depending on factors such as the formulation method, administration method, age, body weight, sex, pathological condition, and the like of the patient, and may be appropriately selected by those skilled in the art. In general, the pharmaceutical composition of the present invention may be administered in an amount of 0.001-100 mg / kg on an adult basis, once to several times a day. However, the dose is not limited in any way to the scope of the present invention.

As another specific use of the present invention, the composition of the present invention can be used as a food composition for preventing or ameliorating depression and anxiety.

The term "improvement" as used in the present invention means all actions that improve symptoms of a disease or disease in an individual.

When the composition of the present invention is used as a food composition, components other than the ether or ranol monomethyl ether, which are conventionally added in food production, such as proteins, carbohydrates, fats, nutrients, seasonings and flavors, May be further included.

Examples of such carbohydrates are monosaccharides, such as glucose, fructose, and the like; Disaccharides such as maltose, sucrose, oligosaccharides and the like; And polysaccharides such as dextrin, cyclodextrin and the like, and sugar alcohols such as xylitol, sorbitol and erythritol. Natural flavorings such as tau martin and stevia extract (e.g., rebaudioside A and glycyrrhizin) and synthetic flavors (saccharin, aspartame, etc.) may be used as flavorings.

The food composition of the present invention may contain various nutrients, vitamins, electrolytes, flavors, colorants, pectic acid and salts thereof, alginic acid and salts thereof, organic acids, protective colloid thickeners, pH adjusting agents, stabilizers, preservatives, glycerin, Alcohols, carbonating agents used in carbonated drinks, and the like. In addition, the food composition of the present invention may contain flesh for the production of natural fruit juices, beverages and vegetable drinks. These components may be used independently or in combination.

There is no particular limitation on the kind of the food. Examples of foods to which the ether or rheol monomethyl ether of the present invention can be added include meat, sausage, bread, chocolate, candy, snack, confectionery, pizza, ramen noodle, other noodles, dairy products including ice- Drinks, tea, drinks, alcoholic beverages, and vitamin complexes, all of which include health foods in a conventional sense.

When the food composition of the present invention is prepared as a drink, citric acid, liquid fructose, sugar, glucose, acetic acid, malic acid, fruit juice and the like may be further added in addition to the ether or ranol monomethyl ether of the present invention.

In addition, the food composition of the present invention may further include food additives, and the suitability of the food additives as food additives is not limited to those specified in the General Rules for Food Additives approved by the Food and Drug Administration, Standards and standards.

Examples of the foodstuffs that have been used in the above food additives include chemical compounds such as ketones, glycine, potassium citrate, nicotinic acid, and cinnamic acid; Natural additives such as crystalline cellulose and guar gum; Sodium L-glutamate preparation; Noodle-added alkaline agent; Preservative formulation; And mixed preparations such as tar coloring agents.

On the other hand, the ether or ranol monomethyl ether of the present invention is a harmless substance having no cytotoxicity. Therefore, the ether or ranol monomethyl ether of the present invention can be safely used for a long period of use, and can be safely used especially for the above-mentioned pharmaceutical or plant composition.

The ether or ranol monomethyl ether of the present invention exhibits an effect of selectively, reversibly and competitively inhibiting the activity of monoamine oxidase A (MAO-A), which is associated with the onset of depression and anxiety, and has little cytotoxicity , Depression and anxiety. ≪ Desc / Clms Page number 2 >

Figure 1 shows the time-dependency of MAO-A inhibition by alternaria monomethyl ether (AME).
Figure 2 shows the recovery of MAO-A inhibitory activity by AME.
Figure 3 shows the line Weibull-Burk plot (A) of inhibition of MAO-A by AME and the second plot (B) of the slope versus inhibitor concentration.

Hereinafter, the present invention will be described in more detail through preparation examples and test examples. These preparations and preparations are only for the purpose of describing the present invention in more detail and that the scope of the present invention is not limited by these preparations and test examples according to the gist of the present invention, It will be obvious to those who have.

Material preparation

Benzylamine, quinolamine, and recombinant human MAO-A and MAO-B were purchased from Sigma-Aldrich (St. Louis, Mo.); MAO-A and MAO-B were stored at -70 C in 100 mM potassium phosphate, pH 7.4, 0.25 M sucrose, 0.1 mM EDTA, and 5% glycerol.

Reference Example 1 Measurement of initial oxidation rate of MAO

The initial oxidation rate of MAO was measured at 25 DEG C in a 1 ml cuvette containing 50 mM sodium phosphate (pH 7.2). The activity of MAO-A was analyzed using 0.2 mM quinolamine as a substrate for 10 minutes at 316 nm and the activity of MAO-B was analyzed at 250 nm for 10 minutes using 0.2 mM benzylamine. The reaction was initiated by adding a substrate to the enzyme mixture. The reaction rate was expressed as the change in absorbance per minute. When this method was used, the K _m values for quinolamine were 0.033 mM and the substrate concentrations were 6.1 X Km, respectively.

Example 1 Preparation and Analysis of Altera and Riol Monomethyl Ether

SNF-008 strain (this strain has 99.6% homology with the strain of Alternaria brassicae ) was isolated from Rheum palmatum stem.

The SNF-008 strain was cultured at 27 DEG C using 1/10 MEA (2.0 g of malt extract) and stirred at 150 rpm for 7 days. After fermentation, the bacterial culture was filtered to separate the culture broth from the mycelium. The mycelial layer was extracted using a 1: 1 mixture of acetone and methanol and the solvent removed under vacuum.

Then, ultrasonic treatment was performed and filtration was performed. A crude extract was obtained after mixing the mycelium extract and the culture solution. The crude extract was fractionated by silica flash column chromatography and eluted with CH ₂ Cl ₂ / CH ₃ OH (100: 1, 50: 1, 20: 1, 1: 100) to give 4 fractions.

Fraction 1 (128.8 g) was loaded on a reversed phase C18 column (Watchers 120 ODS-BP (250 mm ㅧ 10 mm, 5 袖 m), 2.0 ml / min, UV = 254 nm and isocratic condition = 55% CH3CN To give compound 1 (1.3 mg) and compound 2 (1.6 mg).

Compound 1 ¹ H NMR spectral analysis of the two metacoupled aromatic protons set (δ 7.21 (d, 1H, J = 2.1 Hz), 6.75 (d, 1H, J = 2.5 Hz), 6.67 (d, 1H, J = 2.1 Hz), and 6.65 (d, 1H, J = 2.5 Hz)); And two methyl singlets (? 3.61 (s, 3H) and 2.72 (s, 3H)).

¹³ C NMR spectra of Compound 1 showed 14 carbon signals (δ 137.4 (C), 99.1 (CH), 164.2 (C), 99.9 (CH), 165.7 152.2 (CH), 101.2 (CH), 158.1 (C), 117.2 (CH), 138.0 (C), 55.4 (CH3), and 24.5 (CH3).

Compound 1 obtained by comparing and analyzing the result of the 2D NMR spectrum analysis with the previous NMR data was identified as AME having the structural formula of the following formula (1).

[Chemical Formula 1]

¹ H NMR spectral analysis of the compound 2, 2 orthocoupled aromatic protons (δ 8.16 (d , 1H, J = 8.8 Hz), 8.08 (d, 1H, J = 8.7 Hz), 7.16 (d, 1H, J = 8.8 (D, 1H, J = 3.5Hz), 3.60 (m, 1H), and 3.35 (s, 1H) ), And three methine protons (2.99 (m, 1H), 2.78 (m, 1H), and 2.45 (dt, 1H, J = 3.5, 2.0 Hz).

As a result of analyzing the spectrum of the compound 2, it was identified as ATX-II having the structure of the following formula (2).

Example 2 Measurement of MAO-A and MAO-B Inhibitory Activity of AME and ATX-II

MAO-A and MAO-B inhibition of AME and ATX-II. IC ₅₀ values were determined by constructing an S-shaped dose-response curve using residual MAO activity in the presence of various concentrations of inhibitor. Toloxatone and Clorgyline were used as reference inhibitors for MAO-A and MAO-B, respectively. The results are shown in Table 1.

compound IC ₅₀ ([mu] M) SI ^a  MAO-A  MAO-B AME 1.71 ± 0.13 > 80  > 46.8 ATX  > 80  > 80  - Toloxatone  3.92  -  - Chlorgyline  0.0049 ± 0.0005  > 2.0  -

The value is the mean of three experiments + SE. Values for chloroglycine and pargyline were determined after preliminary incubation of the enzyme for 10 minutes.

SI ^a (selectivity index) is the ratio of IC ₅₀ (MAO-B) / IC ₅₀ (MAO-A).

As shown in Table 1 above, AME strongly inhibited MAO-A with an IC ₅₀ value of 1.71 μM, but did not effectively inhibit MAO-B. AME was more selective for MAO-A than MAO-B with an SI value greater than 46.8.

On the other hand, ATX-II had an inhibitory effect on MAO-B or MAO-A with an IC ₅₀ value of less than 80 μM.

Example 3 Measurement of Time-Dependent Inhibitory Activity of MAO-A according to Pre-Culture Time of AME

Reversibility of MAO-A inhibition by AME was investigated by testing the time-dependence of inhibition and testing for restoration of inhibitory activity. Specifically, after AME at 25 ° C and various pretreatment (0, 1, 2, 5, 10, 15, and 30 minutes), the remaining activity was measured using 2.0 mM quinolamine. AME was used at a concentration of 8.0 μM (4.7 × IC ₅₀ ), which was about twice as high as its IC ₅₀ value. Activity was assayed at 316 nm for 10 minutes in the presence of 2.0 mM quinolamine as substrate. Control groups were reacted in the presence of substrate without inhibitor.

Figure 1 shows the time-dependency of MAO-A inhibition by AME. As shown here, the pre-incubation time tended to be somewhat lower up to 5 minutes, but the inhibition effect of AMO on MAO-A activity was not different according to the pre-incubation time. In other words, it was found that the MAO-A inhibition of AME occurs immediately regardless of the pre-incubation time.

In addition, restoration of the inhibitory activity was performed by modifying the existing technique. AME (100 IC ₅₀ ) was incubated with MAO-A for 10 minutes and then diluted 100-fold (1.0 X IC ₅₀ ). For comparison, chlorgyline was used as an irreversible MAO-A inhibitor.

Figure 2 shows the recovery of MAO-A inhibitory activity by AME. Where I is the reaction with the substrate in the presence of the inhibitor (1.0 X IC ₅₀ ), U + D is the 100-fold diluted enzyme mixture (final of the inhibitor Concentration 1.0 X IC ₅₀ ).

As a result, the recovery activity of AME (51.2%) in the diluted condition was almost the same (97.0%) as in the undiluted condition (52.8%). On the other hand, in the case of chlorgyline, the recovery activity was reduced from 45.0% to 24.9% (55.3%). These results indicate that AMEr is not an irreversible inhibitor.

Example 4 Measurement of MAO-A Inhibitory Activity by AME Concentration or Substrate Concentration

Recombinant human MAO-A inhibition mechanics by AME were tested using different concentrations of substrate and inhibitor. The inhibition pattern and K _i values were measured using a Lineweaver-Burk plot.

Figure 3 shows the line Weibull-Burk plot (A) of inhibition of MAO-A by AME and the second plot (B) of the slope versus inhibitor concentration. Quinolamine was used as a substrate at five different concentrations (0.02-0.5 mM).

The catalytic rate of MAO-A was measured in the presence or absence of inhibitor at five different substrate concentrations (0.2-5.0 mM). The line weir plot for inhibition of MAO-A by AME was linear and crossed the y-axis (Fig. 4), indicating that AME is a mixed inhibitor of MAO-A. From the second plot of slope to inhibitor concentration, the K _i value for MAO-A inhibition by AME was determined to be 0.34 ± 0.04 μM, which is slightly higher than that of harman (0.26 μM) (3.34 [mu] M) and anthiactin A (1.84 [mu] M). From this, it can be seen that AME is the most potent selective reversible MAO-A inhibitor.

The ether or ranol monomethyl ether of the present invention exhibits an effect of selectively, reversibly and competitively inhibiting the activity of monoamine oxidase A (MAO-A), which is associated with the onset of depression and anxiety, and has little cytotoxicity , Depression and anxiety. ≪ Desc / Clms Page number 2 >

Claims (4)

A pharmaceutical composition for preventing or treating depression and anxiety, which comprises an alternaria monomethyl ether or a pharmaceutically acceptable salt thereof as an active ingredient. The method according to claim 1,
The pharmaceutical composition for preventing or treating depression and anxiety according to claim 1, wherein the ether or ranol monomethyl ether is isolated from Alternaria brassicae . Wherein the food composition comprises at least one selected from the group consisting of aldehyde monohydrate, The method of claim 3,
The food composition for preventing or ameliorating depression and anxiety according to claim 1, wherein the ether or ranol monomethyl ether is isolated from Alternaria brassicae .

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