KR101928346B1 - Novel mangiferin derivatives and composition having anti-inflammatory activity comprising the same - Google Patents

Abstract

[화학식 2]

[화학식 3]

TECHNICAL FIELD The present invention relates to a maculopyrine derivative which inhibits nitric oxide production and an anti-inflammatory composition comprising the maculopyrin derivative and an effective ingredient thereof. The present invention has an excellent effect of inhibiting the NO production of macrophages and has no side effects.
The macroreticular perine derivative of the present invention is characterized in that it is produced by irradiating a mixed solution obtained by mixing maculopyrine with radiation, wherein the maculiperidine derivative is represented by the following Chemical Formula 1, Chemical Formula 2, Chemical Formula 3 .
[Chemical Formula 1]

(2)

(3)

Description

TECHNICAL FIELD [0001] The present invention relates to a macrorbitopherin derivative which inhibits nitric oxide production and an antiinflammatory composition comprising the macrorbitopiperin derivative as an effective ingredient. BACKGROUND ART < RTI ID = 0.0 >

TECHNICAL FIELD [0001] The present invention relates to a maculopyrine derivative inhibiting nitric oxide production and an anti-inflammatory composition containing the same.

Inflammatory reactions are caused by various inflammatory mediators and immune cells in the local blood vessels and body fluids when the tissue (cell) is damaged or exposed to external infectious agents (bacteria, fungi, viruses, various kinds of allergens) Inflammatory mediator secretion, body fluid infiltration, cell migration, tissue destruction, erythema, edema, and fever.

In general, the inflammatory reaction removes external infectious agents and regenerates damaged tissues, thereby inducing the function of restoring biological functions. However, when the antigen is not continuously removed or the specific internal substance causes the inflammatory reaction to persist, it becomes an irritable disease or chronic inflammation, and it is also a factor in the treatment process such as transfusion, drug administration, or organ transplantation.

It is known that nitric oxide (NO) is formed by nitric oxide synthase (NOS) and plays an important role in mediating the inflammatory reaction .

Nitric oxide is a reactive nitrogen having a short half-life and exhibits various physiological functions such as relaxation of blood vessels, neurotransmission, blood coagulation, inflammation reaction, and defense of the host immune system against tumor cells and parasites. However, Or high cytotoxicity in the presence of high concentrations.

There are several kinds of NOS (nitric oxide synthase), which are cNOS (constitutive NOS: a protein existing in the cell, which is activated by some stimulus), brain nOS (brain NOS) The present nNOS (neuronal NOS) and the eNOS (endothelial NOS) in the blood vessel system are always expressed in the body at a certain level. The small amount of NO produced by them plays an important role in maintaining normal body homeostasis, such as inducing neurotransmission or vasodilation.

On the other hand, NO, which is rapidly over-produced by various cytokines or external stimulants induced by iNOS (inducible NOS), is known to cause cytotoxicity and various inflammatory responses.

Macrophages are known to play a very important role in cell damage and inflammatory responses. Especially, macrophages express LPS (lipopolysaccharide), tumor necrosis factor-α (TNF-α), interleukin-1β (IL- And interferon-γ (IFN-γ), they secrete reactive oxygen species such as superoxide and hydrogen peroxide (H ₂ O ₂ ). By these, macrophage activation is promoted and microorganisms It shows functions such as sterilization effect and cell lysis function.

On the other hand, stimulated macrophages express nitric oxide (NO ₂ ^- ) and nitrate (NO ₃ ^- ) and nitrate (NO ₃ ^- ) through the expression of inducible nitric oxide synthase (iNOS) ), Thereby increasing the inflammatory response and causing cell and tissue damage.

Thus, treatment of inflammation is an important target to inhibit excessive production of NO in macrophages.

To date, nonsteroidal anti-inflammatory drugs (NSAIDs) have been used as anti-inflammatory agents. Nonsteroidal antiinflammatory agents are temporarily effective for reducing inflammation and relieving pain. However, if taken for a long time, it may lead to secondary anemia due to gastrointestinal peptic ulcer bleeding, inhibition of platelet function, induction of labor, side effects on kidney, It causes severe side effects such as hypersensitivity reaction.

Therefore, in order to overcome the side effects of conventional anti-inflammatory drugs, there is a strong demand for the development of anti-inflammatory agents derived from natural materials with less adverse effects on the human body and excellent anti-inflammatory effects.

Korean Patent Laid-Open Publication No. 10-2015-0130207 discloses an anti-inflammatory drug using natural products, which contains aloe flower extract as an active ingredient, thereby eliminating reactive oxygen species (ROS) (COX-2, cyclooxygenase) by inhibiting the inducible nitric oxide synthetase (iNOS) and the antioxidant, anti-inflammatory or immunomodulating activity I am suggesting.

Korean Patent No. 10-0991398 discloses a method for inhibiting the production of nitrogen oxides (NO), inhibiting the production of prostaglandin E2 (PGE2), inhibiting the expression of inducible nitric oxide synthase (iNOS), and inhibiting the proinflammatory cytokine IL- An anti-inflammatory composition containing an extract of Desmarestia viridis, which is a seaweed having an anti-inflammatory effect through its expression-inhibiting activity, as an active ingredient.

On the other hand, manganese perine is a representative C-glucosidic xanthone found in plants belonging to the species of Anarcardiaceae, Gentianaceae, Clusiaceae and Asparagaceae, and is known to contain high content in the shell of mango belonging to Anarcardiaceae.

Manganese perrine has been identified as having anti-obesity, antioxidant, anti-diabetic, antiviral and immunomodulatory properties from previous studies, and is still being studied. However, the present study focuses on confirming the new efficacy of manganese perrin, and there has been no study to confirm the production of derivatives by the structural modification of manganese perrin and its effect.

The present inventors have found a novel derivative of mandarin fermented by irradiating manganese perrine and a mixed solution with radiation as a part of the development of an anti-inflammatory agent and an NO inhibitor derived from natural materials, and the derivative is RAW 264 induced by inflammation induced by LPS (lipopolysaccharide). 7 < / RTI > macrophages, leading to the present invention.

Korean Patent Laid-Open No. 10-2015-0130207 (composition having antioxidant, anti-inflammatory or immunomodulating activity containing aloe flower extract as an active ingredient) Korean Patent No. 10-0991398 (composition for antiinflammation containing an extract of Aspergillus oryzae as an active ingredient)

It is an object of the present invention to solve the above problems and to provide a novel maculopyrine derivative which is excellent in inhibiting the NO production of macrophages and has no side effects and an antiinflammatory composition containing the maculopyrin derivative as an active ingredient.

In order to solve the above-mentioned problems, the present invention provides the following compounds represented by the following formulas (1), (2) and (3)

[Chemical Formula 1]

(2)

(3)

The above-mentioned maculopyrine derivative is characterized in that it is produced by irradiating a mixed solution obtained by mixing maculopyrine represented by the following formula (4) with radiation.

[Chemical Formula 4]

The radiation may be any one selected from the group consisting of gamma rays, electron beams, UV and X-rays, and the radiation may be irradiated at a dose of 10 to 200 kGy.

The compounds of formula (1), (2) and (3) are characterized in that they inhibit the production of NO (nitric oxide) and have anti-inflammatory action.

The mixture of buckwheat ferrine according to the present invention may be prepared by directly purchasing buckwheat perinein reagent sold in the market and mixing with an organic solvent or using a product isolated from a plant extract containing buckwheat perlin such as mango extract.

The mango extract may contain water or an organic solvent extract, preferably an organic solvent extract, a crude extract or a concentrate thereof.

The organic solvent is preferably at least one selected from the group consisting of lower alcohols having 1 to 4 carbon atoms, hexane, ethyl acetate, dichloromethane, ether, chloroform and acetone.

The lower alcohol having 1 to 4 carbon atoms is preferably at least one selected from the group consisting of methanol, ethanol, propanol, isopropanol, butanol, and n-butanol, and the lower alcohol having 1 to 4 carbon atoms includes anhydride or hydrated alcohol .

The alcohol, such as ethanol or isopropanol, preferably contains 1 to 100% (v / v), preferably 30 to 100%, more preferably 70 to 100%, still more preferably 90%, 95%, 100 % Alcohol. When butanol is used as an extraction solvent, it is preferable to use water-saturated butanol (75 to 85% aqueous solution of butanol).

The mango extract may include not only extracts or fractions derived from the above-mentioned extraction solvent, but also extracts obtained through ordinary purification processes. For example, various additional purification methods, such as separation using an ultrafiltration membrane with a constant molecular weight cut-off value, separation by various chromatographies (made for separation by size, charge, hydrophobicity or affinity) The fractions obtained through this step are also included in the fractions of the present invention.

In order to solve the above problems, the present invention provides an anti-inflammatory composition comprising a macrorbitopherin derivative represented by any one of the above-mentioned Formulas (1), (2) and (3) as an active ingredient.

The anti-inflammatory composition is characterized by its anti-inflammatory action as a mechanism of inhibiting the production of NO (nitric oxide).

The anti-inflammatory composition may be, but is not limited to, a pharmaceutical composition or a potentially health food composition.

The pharmaceutical composition of the present invention can be prepared using a physiologically acceptable adjuvant. Examples of the adjuvant include excipients, disintegrants, sweeteners, binders, coating agents, swelling agents, lubricants, lubricants, have.

In addition, one or more pharmaceutically acceptable carriers may be optionally formulated into pharmaceutical compositions.

The pharmaceutical composition may be in the form of granules, powders, tablets, coated tablets, capsules, suppositories, liquids, syrups, juices, suspensions, emulsions, drops or injectable solutions. For example, for formulation into tablets or capsules, the active ingredient may be combined with an oral, non-toxic pharmaceutically acceptable inert carrier such as ethanol, glycerol, water, and the like.

Also, if desired or necessary, suitable binders, lubricants, disintegrants and coloring agents may also be included as a mixture. Suitable binders include, but are not limited to, natural sugars such as starch, gelatin, glucose or beta-lactose, natural and synthetic gums such as corn sweeteners, acacia, tracker candles or sodium oleate, sodium stearate, magnesium stearate, sodium Benzoate, sodium acetate, sodium chloride, and the like. Disintegrants include, but are not limited to, starch, methyl cellulose, agar, bentonite, xanthan gum and the like.

Acceptable pharmaceutical carriers for pharmaceutical compositions of the present invention include those suitable for sterilization and in vivo such as saline, sterilized water, Ringer's solution, buffered saline, albumin injection solution, dextrose solution, maltodextrin solution, glycerol, , And if necessary, other conventional additives such as an antioxidant, a buffer, and a bacteriostatic agent may be added.

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. Further, it can be suitably formulated according to each disease or ingredient, using the method disclosed in Remington's Pharmaceutical Science, Mack Publishing Company, Easton PA as an appropriate method in the field.

The dosage of the pharmaceutical composition according to the present invention should be a pharmaceutically effective amount. A " pharmaceutically effective amount " means an amount sufficient to prevent or treat a disease at a reasonable benefit / risk ratio applicable to medical treatment, and the effective dosage level will vary depending on factors such as the formulation method, the condition and weight of the patient, The severity of the disease, the mode of administration, the route and time of administration, the rate of excretion, the responsiveness, and the like. Effective amounts may vary depending on the route of treatment, the use of excipients, and the likelihood of use with other agents, as will be appreciated by those skilled in the art.

According to still another aspect of the present invention, there is provided a health functional food composition comprising a mangiferin derivative as an active ingredient.

The health functional food is a food emphasizing the biological control function of food, and is added with added value so as to function and manifest to a specific purpose using physical, biochemical and biotechnological methods. These functional food ingredients are designed and manufactured so that the body control functions related to the regulation of the body defense and the body rhythm, the prevention and the recovery of the disease are sufficiently exhibited to the living body, and the food can be supplemented with food, a sweetener or a functional ingredient ≪ / RTI >

Examples of the health functional foods according to the present invention include various foods such as beverages, gums, tea, vitamin complexes, and health supplement foods. The drinks include natural fruit juices, fruit juice drinks, . The food composition of the present invention can be prepared in the form of tablets, tablets, granules, powders, capsules, liquid solutions, rings and the like according to known production methods.

In addition, the health functional food of the present invention may further contain various conventional flavors, natural carbohydrates and the like. Flavors include natural flavors such as tau Martin and stevia extract, and synthetic flavors such as saccharin and aspartame.

The above-mentioned natural carbohydrates may include polysaccharides such as disaccharides such as glucose, monosaccharide such as fructose, maltose, sucrose, dextrin, cyclodextrin, sugar alcohols such as xylitol, sorbitol, erythritol, and the like .

In addition, the food composition of the present invention may further contain various nutrients, vitamins, minerals (electrolytes), flavors such as synthetic flavors and natural flavors, coloring agents, heavy stabilizers (cheese, chocolate etc.), pectic acid and its salts, Salt, organic acids such as anhydrous citric acid, protective colloid thickener, pH adjusting agent, stabilizer, preservative, glycerin, alcohol, carbonating agent used in carbonated drinks, natural fruit juice, fruit juice etc. Lt; RTI ID = 0.0 > additive < / RTI > These additives may be used independently or in combination.

As described above, the novel maculopyrine derivative according to the present invention and the anti-inflammatory composition containing the maculopyrine derivative according to the present invention have an excellent effect of inhibiting macrophage NO production and no side effects.

Hereinafter, preferred embodiments of the present invention will be described in detail with reference to the accompanying drawings and diagrams. The detailed description of the functions and configurations of the present invention will be omitted if it is determined that the gist of the present invention may be unnecessarily blurred.

Hereinafter, the respective compounds represented by the formulas (1), (2), (3) and (4) will be referred to as compounds 1, 2, 3 and 4 in order.

Example  1. Isolation of Compound

A sample solution containing 500 mg of manganese perlein (Compound 4) in 300 ml of methanol was irradiated at 50 kGy (absorbed dose) and dried.

The dried sample was separated by column chromatography using a YMC GEL ODS AQ 120-50S column (1.5 cm i.d. x 40 cm).

The solvent was used in a stepwise gradient manner while increasing the amount of methanol with a mixed solvent of methanol and water. Compound 1 (40.7 mg, t _R 7.2 min), Compound 2 (5.9 mg, t _R 9.5 min) compound _{3 (14.1 mg, t R 10.3} min) was isolated pure.

Reversed-phase HPLC analysis was performed using a YMC-Pack ODS A-302 column (4.6 mm i.d. x 150 mm; YMC Co., Ltd.).

The solvent system consisted of linear gradient MeCN-0.1% HCOOH / H ₂ O (measurement: UV 280 nm; flow rate: 1.0 ml / min; temperature 40 ° C).

Example  2. Identification of compounds

Compound 1 was isolated as a yellow amorphous powder and was characterized by ¹³ C-NMR and HRFABMS measurements ( m / z 467.1188 [MH] ^- (calcd. For C ₂₁ H ₂₃ O ₁₂ , 467.1190) ₂₁ H ₂₄ O ₁₂ .

The maximum absorption wavelength at 220 and 260 nm of the UV spectrum was estimated to be due to the xanthene nucleus.

Table 1 below shows ¹ H NMR and ¹³ C-NMR data of Compound 1, Compound 2 and Compound 3.

As shown in Table 1, ¹ H NMR spectrum of the compound 1 is a _{δ H 6.93 (1H, s,} H-8), 6.41 (1H, s, H-5) and 6.06 (1H, s, H- 4) in Showed three proton signals, and Compound 1 was found to belong to an aromatic compound having a xanthene moiety.

Six oxygen-bearing protons were observed at δ _H 4.91 (1H, d, J = 10.2 Hz, H-1 ') at δ _H 3.82-3.41 and the presence of glucopyranosyl unit there was.

The ¹ H NMR spectrum of compound 1 showed the following characteristics: δ _H 4.43 (1H, d, J = 11.4 Hz, H-11) 11.4 Hz, H-11) and 3.81 (1H, d, J = 11.4 Hz, H-12).

As shown by ¹ H NMR, the ¹³ C NMR and HSQC spectra of Compound 1 showed that two hydroxymethyl groups (H-11 (隆 c 66.7) and H-12 (隆 c 67.7) in place of the ketone moiety at C- Except for the presence of the parent compound.

As shown in Formula 1, the HMBC spectrum of Compound 1 showed H-11 / C-8a, 9, 9a and H-12 / C-8a, 9 and 9a correlations and hydroxymethyl residues were found at C-9 .

The spectroscopic analysis was performed by Extended 2D NMR techniques and the new compound 1 showed that it had two hydroxymethyl groups modified by radiation. Based on the above results, Compound 1 was named mangiferdiol.

HRFABMS of compounds 2 and 3 showed identical molecular ion peaks at m / z 451.1241 [MH] (calcd. For C ₂₁ H ₂₃ O ₁₁ , 451.1240). In addition, it has been confirmed that the compounds 2 and 3 have a structural formula of C ₂₁ H ₂₄ O _{11 with} one oxygen atom less than Mangiferdiol.

¹ H NMR and ¹³ C NMR spectral data showed that compounds 2 and 3 were similar to compound 1, but compound 1 showed a hydroxymethyl signal at H-12 whereas compounds 2 and 3 showed a methyl signal at H-12 It showed existence.

The position of the methyl group was confirmed by the HMBC correlations of H-12 / C-8a, 9, 9a. Compound 3, which is the relative stereostructure of Compound 2 and its epimer, was confirmed by comparing the degree of non-linearity.

Based on the above results, compounds 2 and 3 were named mangiferinol and isomangiferinol, respectively, as a new radiolytic product of macelferrin.

Mangiferdiol (Compound 1) : Yellow amorphous powder, [a] ²⁵ _D -34.0 (c 0.1, MeOH); UV (MeOH)? Max (log?): 220 (3.09), 260 (1.03) nm; ¹ H and ¹³ C NMR, shown in Table 1; FABMS m / z 467 [MH] ^- , HRFABMS m / z 467.1188

^{[MH] - (. Calcd for} C 21 H 23 O 12, 467.1190).

Mangiferinol (Compound 2) : Yellow amorphous powder, [a] ²⁵ _D -42.1 (c 0.1, MeOH); UV (MeOH)? Max (log?): 221 (3.24), 260 (1.09) nm; ¹ H and ¹³ C NMR, shown in Table 1; FABMS m / z 451 [MH] ^- , HRFABMS m / z 451.1241

^{[MH] - (. Calcd for} C 21 H 23 O 11, 451.1240).

Isomangiferinol (Compound 3) : yellow amorphous powder, [a] ²⁵ _D +53.0 (c 0.1, MeOH); UV (MeOH)? Max (log?): 221 (3.23), 260 (1.08) nm; ¹ H and ¹³ C NMR, shown in Table 1; FABMS m / z 451 [MH] ^- , HRFABMS m / z 451.1241

^{[MH] - (. Calcd for} C 21 H 23 O 11, 451.1240).

Example  3. Measurement of NO inhibitory activity

NO production inhibitory activity was measured in LPS-stimulated RAW 264.7 cells of compound 1, compound 2, and compound 3, which were purely isolated from manganese perine (compound 4).

Table 2 shows the NO production inhibitory activity of the compounds 1 to 3.

All experimental results were performed more than 3 times and Mean ± SD. , And it was judged to be significant when the p-value was less than 0.05. Aminoguanidine was used as a positive control.

In this new compound, the modified kappelegine derivative (compound 1) (with two hydroxymethyl groups instead of the ketone moiety at C-9) showed an IC ₅₀ value of 47.1 ± 1.7 μM, indicating excellent NO production inhibition.

On the other hand, compound 2 and its C-9 epimers of compound 3 showed a somewhat lower NO production inhibitory activity by showing respectively 76.0 ± 2.0 a IC ₅₀ value of 82.0 ± 2.1 μM.

These results suggest that the number of hydroxymethyl groups at the C-9 ketone position in the mangiferin skeleton has a positive effect on the inhibition of NO production.

Radiative decomposition of methanol in the presence of oxygen causes various free radicals such as methoxy (CH3O ·), hydroxyalkyl (· CH ₂ OH), hydrogen (H ·), superoxide anion (O ₂ ·), and peroxyl (HOO ·) And expected to lead to the production of oxygen containing compounds, and the results showed that the free radicals generated by methanol radiolysis affect the synthesis of new compounds.

While the present invention has been particularly shown and described with reference to exemplary embodiments thereof, it is clearly understood that the same is by way of illustration and example only and is not to be taken as limiting the scope of the present invention. The present invention can be variously modified or modified. The scope of the invention should, therefore, be construed in light of the claims set forth to cover many of such variations.

Claims (9)

A mangiferin derivative represented by any one of the following formulas (1), (2) and (3)
[Chemical Formula 1]

(2)

(3)

The method according to claim 1,
The maculopyrine derivative
Characterized in that it is produced by irradiating a mixed solution obtained by mixing a mandarin perine represented by the following formula (4)
Manganese perine derivative.

[Chemical Formula 4]

3. The method of claim 2,
The mortar ferrine mixed solution
Characterized in that it is a mixture of geranium perlein and an organic solvent having 1 to 4 carbon atoms
Manganese perine derivative.
3. The method of claim 2,
The radiation
Gamma ray, electron beam, UV and X-ray.
Manganese perine derivative.
5. The method of claim 4,
The radiation
Is irradiated at a dose of 10 to 200 kGy
Manganese perine derivative.
The method according to claim 1,
The maculopyrine derivative
Characterized in that production of NO (nitric oxide) is inhibited
Manganese perine derivative.
1. An anti-inflammatory composition comprising a mangiferin derivative represented by any one of the following general formulas (1), (2) and (3) as an active ingredient.
[Chemical Formula 1]

(2)

(3)

8. The method of claim 7,
The anti-
Characterized in that production of NO (nitric oxide) is inhibited
An anti-inflammatory composition comprising a mangiferin derivative as an active ingredient.
8. The method of claim 7,
The anti-
Characterized in that it is a pharmaceutical composition or a potentially health food composition
An anti-inflammatory composition comprising a mangiferin derivative as an active ingredient.