KR101839313B1 - Antidiabetic composition comprising radiation-treated rutin derivative as effective component - Google Patents

Abstract

The present invention relates to an antidiabetic composition containing a rutin derivative treated with radiation as an active ingredient. The antidiabetic composition of the present invention can be usefully utilized for a material of a diabetic medicine or health functional foods for preventing or alleviating diabetes since a rutin derivate hydroxymethylated by irradiating with radiation, more specifically a gamma ray, represents a batter α-glucosidase deactivating effect than that of rutin which is not irradiated with the gamma ray.

Description

[0001] The present invention relates to an antidiabetic composition containing a radiation-treated rutin derivative as an active ingredient,

The present invention relates to a composition for antidiabetic use containing a rutin derivative hydroxymethylated by treating with radiation as an active ingredient.

Diabetes is a type of metabolic disease that lacks insulin secretion or does not function normally. It is characterized by hyperglycemia with elevated blood glucose concentration. Hyperglycemia causes various symptoms and signs, and it excretes glucose from the urine.

Diabetes mellitus is divided into type 1 and type 2, type 1 diabetes was formerly called 'child diabetes' and is caused by failure to produce insulin at all. Type 2 diabetes is characterized by relatively insufficient insulin, insulin resistance that lowers insulin function that lowers blood sugar, and cells do not burn glucose effectively. Type 2 diabetes accounts for more than 90% of all diabetes, and type 2 diabetes is rapidly increasing as the obesity population increases due to low lifestyle and westernized diets.

The most effective treatment of type 2 diabetes is to achieve optimal blood glucose levels after meals and delayed absorption of monosaccharides through the inhibition of? -Glucosidase activity is presented as a beneficial strategy for the treatment of type 2 diabetes have.

Radiation Technology is a technique for irradiating a radiation source such as a gamma ray, an electron beam, or an X-ray or an energy generated from a radiation generator with an irradiation dose of 0.01 kGy to 200 kGy. In recent years, Research on the development of high value added biomaterials by enhancing the physiological activity of the effective ingredient extraction rate and structural change of the active ingredient by applying the irradiation technique is underway and it is known that the increase of activity of a specific substance, Studies have also been reported to increase or change the content, but research on the structural transformation of the routine due to irradiation has not been conducted.

On the other hand, a rutin is a glycoside with a sugar attached to a flavonoid and a brownish component. It was found in the rutaceae plants of Udonthwa, and later the flower bud of Sophora japonica , buckwheat of Fagopyrum esculentum ) were also isolated from many kinds of plants. It acts to strengthen capillaries, and can be used to prevent hemorrhage, radiation damage, hemorrhagic diseases, and the like.

Korean Patent No. 1628950 discloses a composition for preventing and treating complications due to pre-diabetes, diabetes, or diabetes comprising Rg3, rutin or a combination thereof as an active ingredient. However, the radiation-treated routine of the present invention, More specifically, it has not been disclosed about a composition for antidiabetic use containing a rutin derivative hydroxymethylated by gamma irradiation as an active ingredient.

The present invention provides a composition for anti-diabetes comprising a rutin derivative hydroxymethylated by irradiation with gamma rays as an active ingredient, and is characterized in that an α-glucosidase activity And that the inhibitory effect is enhanced as compared with the routine in which the gamma ray is not irradiated, thereby completing the present invention.

In order to solve the above problems, the present invention provides a pharmaceutical composition for preventing or treating diabetes mellitus comprising the lutein derivative represented by the general formula (1), (2) or (3) or a pharmaceutically acceptable salt thereof as an active ingredient.

The present invention also provides a health functional food composition for preventing or ameliorating diabetes, which comprises the lutin derivative represented by the general formula (1), (2) or (3) or a pharmaceutically acceptable salt thereof as an active ingredient.

The present invention relates to a composition for anti-diabetes containing radiation-treated rutin as an active ingredient, wherein a hydroxymethylated rutin derivative by gamma irradiation is a? -Glucosidase Activity inhibitory effect, it can be utilized as a therapeutic agent for diabetes or as a material for health functional foods for preventing or ameliorating diabetes.

Fig. 1 shows the structure of the novel compounds 1 to 3, wherein (a) shows the interaction relationship between hydrogen and carbon as a result of the HMBC of the compounds 1 to 3, and (b) 1 < / RTI > and 2, which show the interaction relationship between the working hydrogen.

The present invention relates to a pharmaceutical composition for preventing or treating diabetes mellitus comprising, as an active ingredient, a lutein derivative represented by the following general formula (1), (2) or (3) or a pharmaceutically acceptable salt thereof.

The lutein derivative is produced by irradiating the lutein with radiation, preferably by irradiating gamma ray to form a lutein derivative, but the present invention is not limited thereto.

The gamma rays are preferably irradiated so that the absorbed dose is 20 to 30 kGy, more preferably the absorbed dose is 25 kGy, but the present invention is not limited thereto.

In one embodiment of the present invention, the lutein derivative has an inhibitory activity of? -Glucosidase enhanced as compared to that of lutein, but is not limited thereto.

The pharmaceutical composition of the present invention may further comprise a pharmaceutically acceptable carrier, excipient or diluent in addition to the active ingredient. Such a carrier is usually used at the time of formulation, and includes lactose, dextrose, sucrose, sorbitol, mannitol, Starch, acacia gum, calcium phosphate, alginate, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water, syrup, methylcellulose, methylhydroxybenzoate, propylhydroxybenzoate, talc, Magnesium stearate, 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.

The pharmaceutical composition of the present invention can be prepared by any one of formulations selected from capsules, powders, granules, tablets, suspensions, emulsions, syrups and aerosols, but is not limited thereto.

The appropriate dosage of the pharmaceutical composition according to the present invention may vary depending on such factors as the formulation method, the administration method, the patient's age, body weight, sex, pathological condition, food, administration time, administration route, excretion rate, .

The pharmaceutical composition of the present invention may be administered orally or parenterally. In the case of parenteral administration, the composition may be administered topically to the skin, intravenously, subcutaneously, intramuscularly, intraperitoneally, or transdermally.

The present invention also relates to a health functional food composition for preventing or ameliorating diabetes comprising the lutein derivative represented by the above formula (1), (2) or (3) or a pharmaceutically acceptable salt thereof as an active ingredient.

The health functional food composition may be prepared in the form of powders, granules, pills, tablets, capsules, candies, syrups or beverages, but is not limited thereto.

When the health functional food composition of the present invention is used as an additive, the health functional food composition may be added as it is, or may be used together with other food or food ingredients, and may be suitably used according to a conventional method. The amount of the active ingredient can be suitably used depending on its use purpose (prevention or improvement). Generally, the health functional food composition of the present invention is added in an amount of not more than 15 parts by weight, preferably not more than 10 parts by weight based on the raw material, when the food or beverage is produced. However, in case of long-term ingestion for health, the amount may be less than the above range, and since there is no problem in terms of safety, the active ingredient may be used in an amount exceeding the above range.

There is no particular limitation on the kind of the health functional food. Examples of foods to which the health functional food composition can be added include beverages, tea, spices, gum, confectionery, capsules, powders, suspensions, and pills, all of which include health foods in a conventional sense.

In addition, the health functional food composition of the present invention can be produced as a food, particularly a functional food. The functional food of the present invention includes components that are ordinarily added in food production, and includes, for example, proteins, carbohydrates, fats, nutrients, and seasonings. For example, in the case of a drink, a natural carbohydrate or a flavoring agent may be included as an additional ingredient in addition to the active ingredient. The natural carbohydrate may be selected from the group consisting of monosaccharides (e.g., glucose, fructose, etc.), disaccharides (e.g., maltose, sucrose etc.), oligosaccharides, polysaccharides (e.g., dextrin, cyclodextrin, , Xylitol, sorbitol, erythritol, etc.). The flavoring agent may be a natural flavoring agent (e.g., tau Martin, stevia extract, etc.) and a synthetic flavoring agent (e.g., saccharin, aspartame, etc.).

In addition to the above health functional food composition, it is also possible to use various nutrients, vitamins, electrolytes, flavors, colorants, pectic acids and salts thereof, alginic acid and salts thereof, organic acids, protective colloid thickening agents, pH adjusting agents, stabilizers, preservatives, glycerin, A carbonating agent used in beverages, and the like. Although the proportion of the above-mentioned ingredients to be added is not critical, it is generally selected in the range of 0.01 to 0.1 part by weight based on 100 parts by weight of the health functional food composition of the present invention.

Hereinafter, the present invention will be described in more detail with reference to Examples. It is to be understood by those skilled in the art that these embodiments are merely illustrative of the present invention and that the scope of the present invention is not limited thereto.

Example  1. Irradiation of gamma-rays and separation of lutein derivatives for routine

Cobalt-60, which has an activity of about 215 kCi at a dose rate of 25 kGy per hour, was used as a gamma-ray source to generate a novel lutein derivative by irradiating gamma rays to the routine.

A solution of 1 g of rutin dissolved in 1 liter of methanol was irradiated with a gamma ray to a dose of 25 kGy. The irradiated routine was dried immediately and used as a test sample. shifted products.

The sample was run on a Tolryopearl HW-40 column (coarse grade; 2.8 cm id x 32 cm) with water containing constantly increasing methanol and a step gradient on a YMC GEL ODS AQ 120-50 S column (1.5 cm id x 36 cm) ( T _R 10.3 min), 9.8 mg ( t _R 11.6 min) of the new compound 2 and 1.7 mg ( t _R 11.6 min) of the new compound 3 were dissolved in 1.7 to give ㎎ (t _R 7.8min).

Example  2. Structural analysis of lutein derivatives produced by gamma irradiation

The structures of the lutein derivatives (new compounds 1 to 3) obtained in Example 1 were analyzed.

High Performance Liquid Chromatography (HPLC) analysis was carried out using an HPLC system equipped with a YMC-Pack ODS A-302 column (4.6 mm id x 150 mm; YMC Co., Ltd.) (v / v) for 15 minutes using acetonitrile (water: UV 280 nm; flow rate: 1.0 ml / min; The nitrile was increased and then linearly gradiented to 100% acetonitrile for 20 minutes.

In addition, 0.5 mg of the novel compounds 1-3 obtained in Example 1 were dissolved in 0.1 ml of 0.5 N HCl, and then heated at 90 캜 for 2 hours for neutralization. The neutralized solution was dried under vacuum, and the residue was dissolved in 0.1 ml of pyridine to which 0.5 mg of L-cysteine methyl ester hydrochloride had been added, and the mixture was heated at 60 ° C for 1 hour. Then, 0.1 ml of o -torylisothiocyanate (0.5 mg) solution dissolved in pyridine was added to the warmed mixture, which was then heated at 60 ° C for 1 hour. The mixture was immediately analyzed by reverse phase chromatography. Chromatographic analysis was performed on a Cosmosil 5C ₁₈ -MS-II column (4.6 mm id x 250 mm; Nacalai Tesque Inc.) in which 25% (w / v) acetonitrile was dissolved in 50 mM phosphoric acid (H ₃ PO ₄ ) (Detection: UV 250 nm; flow rate: 0.8 ml / min; at 40 ° C). Under these assay conditions, certified D / L-glucose and D / L-rhamnose showed HPLC peaks at t _R 17.8 / 16.4 and 15.7 / 30.2 min, respectively.

As a result of the analysis, the novel compound 1 is a yellow amorphous powder and the specific linearity is [?] ²⁵ _D -15.6 ( c 0.1, MeOH). The UV (MeOH) spectrum is λ _max (log ε): 220 (2.02), 290 (1.58) nm and CD (MeOH) is λ _max (Δε): 291 (+9.71) and 331 (-7.43) nm. The m / z value of FABMS is 641 [MH] ^- and the m / z value of HRFABMS is 641.1710 [MH] ^-: and _{(Calculated for C 28 H 33 O} 17 641.1718), molecular formula is C ₂₈ H ₃₄ O _17. ^{The results of 1} H-NMR and ¹³ C-NMR analyzes are summarized in Table 1, and the structure confirmed from the results of HMBC and NOESY of the novel compound 1 is shown in FIG.

The absorption maxima at 290 nm in the UV spectrum are due to dihydroflavonol nuclei and the composition of the sugars is determined by small scale acid hydrolysis and HPLC analysis of phenylisothiocyanate derivatives using an ultraviolet detector D-glucose and L-rhamnose. The presence of the dihydroflavonol skeleton was confirmed by ¹ H-NMR analysis of the following Table 1 (隆_H 5.45 (1H, s, H-3); 隆_H 7.13 (1H, d, J = 1.8㎐, H-2 '), 6.68 (1H, dd, J = 8.4, 1.8㎐, H-6') , and 6.65 (1H, d, J = 8.4㎐, H-5 ') ABX system;隆_H 6.00 (1H, d, J = 1.8㎐, H-6) , and 5.80 (1H, d, J = 1.8㎐, H-8) the two meta -coulped AB system). ¹³ C-NMR spectrum (隆_C The presence of the dihydroflavonol skeleton of this molecule was confirmed with a characteristic C-ring resonance at 197.1 (C-4), 89.3 (C-2) and 76.4 (C-3)隆_H An additional specific signal of two anomeric protons at 4.85 (1H, d, J = 7.8 Hz, H-1 ") and 4.76 (1H, d, J = 1.8 Hz, H- _H 3.97 ~ 3.37 10 of oxygen-bearing signal and a δ _H 1.19 doublet methyl proton of the said support for the presence of residues on the side Rutino molecule. ^The resonance corresponding to the hydroxymethyl substituent at ¹ H-NMR results (隆_H 3.98 (1H, d, J = 12.6 Hz, H-11a) and 3.65 (1H, d, J = 12.6 Hz, H- Respectively. The ¹³ C-NMR and HSQC spectral results were identical to those of the parent routine except for the oxygenated hydroxymethyl (H-11) and oxymetin (H-3) signals. The point of attachment of the hydroxymethyl moiety at C-2 was clearly determined from the HMBC spectrum showing the H-11 / C-2, 3, 1 'correlation. Relative stereostructure of pseudo-axial catechol at C-2 position and pseudo-equatorial sugar moiety at C-3 position in C-ring was determined using NOESY spectrum. The spectrum shows a correlation between H-3 / H-11, 1 ", 2 ', indicating a trans -relationship between the phenyl group and the rutinosyl moiety. The pseudo-equatorial sugar moiety can be discarded by the observed NOESY correlation of H-3 / H-2 'Furthermore, based on the estimated batch, the energy minimization model generated by Chem3D Ultra 10.0 can be compared with the NOESY result The CD (circular dichroism) spectrum showed a positive and negative Cotton effect at 291 and 331 mm, respectively, confirming that the arrangement of the new compound 1 was a 2R, 3S-configuration. 1 radiorutinol with a novel structure.

Further, the novel compound 2 is a yellow amorphous powder and the specific linearity is [?] ²⁵ _D -33.7 ( c 0.1, MeOH). The UV (MeOH) spectrum is λ _max (log ε): 220 (2.03), 290 (1.57) nm and CD (MeOH) is λ _max (Δε): 301 (-7.43) and 332 (+7.51) nm. The m / z value of FABMS is 642 [M] ^- , the m / z value of HRFABMS is 642.1793 [M] ^- (Calculated for C ₂₈ H ₃₄ O ₁₇ : 642.1796) and the molecular formula is C ₂₈ H ₃₄ O ₁₇ . ^{The results of 1} H-NMR and ¹³ C-NMR analyzes are summarized in the following Table 1, and the structure of the novel compounds was analyzed from the results of HMBC and NOESY of the novel compound 2, which are shown in FIG.

The NMR and UV spectral results were nearly identical to those of the novel compound 1, suggesting that it is a diastereomer. The monosaccharide sequence was determined by D-glucose and L-rhamnose in HPLC analysis using C ₁₈ reversed phase column. The position of the aliphatic compound residue was clearly revealed by the HMBC correlation of the interaction of H-11 / C-2, 3, 1 '. The 2,3- trans stereochemistry of the pseudo-equatiral catechol at the C-2 position of the aglycone moiety and the pseudo-axial sugar moiety at the C-3 position is the NOESY correlation of H-3 / H-2 ' And the absolute stereochemistry of 2S, 3R was determined based on the negative and positive Cotton effect at 301 and 332 mm, respectively, in the CD spectrum based on comparison with intrinsic analogs. One isoradiorutinol.

The novel compound 3 is a yellow amorphous powder and has a specific linearity [?] Of ²⁵ _D +21.4 ( c 0.1, MeOH) and a UV (MeOH) spectrum of λ _max (log ε) 1.65) nm. The m / z value of FABMS is 655 [MH] ^- , the m / z value of HRFABMS is 655.1879 [MH] ^- (Calculated for C ₂₉ H ₃₅ O ₁₇ : 655.1874), and the molecular formula is C ₂₉ H ₃₆ O ₁₇ . ^{The results of 1} H-NMR and ¹³ C-NMR analyzes are summarized in the following Table 1, and the structure of the novel compounds was analyzed from the results of HMBC and NOESY of the new compound 3, which are shown in FIG.

The maximum absorption at 290 nm in the UV spectrum is due to dihydroflavonol and the high similarity of the 1D NMR spectrum with the new compounds 1 and 2, including the HMQC, HMBC and NOESY experimental results, Lt; / RTI > Also, the arrangement of monosaccharides was determined by D-glucose and L-rhamnose. Similarities between the results of the ¹ H-NMR and ¹³ C-NMR experiments of the novel compound 3 and the experimental results of the novel compounds 1 and 2 indicated the presence of additional hydroxymethyl functionality and the unsaturation of C-3 and C- Compound 3 is a unique flavonoid having two hydroxymethyl groups in the dihydropyran ring. Based on the HMBC correlation, the hydroxymethyl groups were located at C-2 and C-4. Based on the above results, the new compound 3 was deduced to be a radiorutindiol containing a unique 2H-pyran C-ring system with two hydroxymethyl groups.

¹ H-NMR and ¹³ C-NMR analysis results location The novel compound 1 The new compound 2 The novel compound 3 隆_H ( J in Hz) δ _c , mult. 隆_H ( J in Hz) δ _c , mult. 隆_H ( J in Hz) δ _c , mult. 2 - 89.3 - 89.3 - 82.3 3 5.45 (s) 76.4 5.45 (s) 78.0 - 143.9 4 - 197.1 - 195.9 - 123.7 5 - 162.2 - 161.5 - 154.5 6 6.00 (d, 1.8) 97.3 6.01 (d, 1.8) 96.6 5.86 (d, 2.4) 98.6 7 - 168.2 - 167.4 - 159.3 8 5.80 (d, 1.8) 96.8 5.81 (d, 1.8) 96.0 5.81 (d, 2.4) 97.1 9 - 163.8 - 162.9 - 155.9 10 - 102.4 - 101.3 - 104.6 11a 3.98 (d, 12.6) 67.4 3.92 (d, 12.6) 66.5 4.17 (d, 12.6) 65.8 11b 3.65 (d, 12.6) 3.72 (d, 12.6) 4.10 (d, 12.6) 12a - - - - 5.07 (d, 12.0) 56.9 12b - - - - 4.00 (d, 12.0) One' - 128.9 - 127.8 - 131.0 2' 7.13 (d, 1.8) 115.5 7.11 (d, 1.8) 114.5 7.00 (d, 2.4) 115.3 3 ' - 145.1 - 144.7 - 145.9 4' - 145.9 - 145.1 - 146.1 5 ' 6.65 (d, 8.4) 115.8 6.66 (d, 8.4) 115.0 6.71 (d, 8.0) 115.2 6 ' 6.68 (dd, 8.4, 1.8) 119.7 6.86 (dd, 8.4, 1.8) 118.8 6.80 (dd, 8.0, 2.4) 120.1 One" 4.85 (d, 7.8) 103.8 4.77 (d, 7.8) 105.2 4.59 (d, 7.8) 100.8 2" 3.50 (m) 77.0 3.53 (m) 76.2 3.50 (m) 76.0 3 " 3.37 (t, 9.6) 73.5 3.46 (t, 9.6) 74.2 3.44 (t, 9.6) 73.6 4" 3.52 (m) 73.4 3.51 (m) 75.5 3.47 (m) 74.2 5 " 3.57 (m) 76.7 3.54 (m) 76.2 3.34 (m) 77.4 6 "a 3.97 (dd, 12.0, 2.4) 67.1 4.09 (dd, 12.0, 2.4) 67.1 3.85 (m) 71.7 6 "b 3.73 (dd, 12.0, 4.8) 3.50 (dd, 12.0, 4.8) 3.70 (m) One"' 4.76 (d, 1.8) 101.7 4.73 (d, 1.2) 100.8 4.78 (d, 1.2) 101.1 2"' 3.85 (dd, 3.6, 1.8) 71.4 3.94 (dd, 3.6, 1.8) 70.3 3.88 (m) 71.2 3 "' 3.69 (m) 69.2 3.66 (m) 68.5 3.55 (m) 67.6 4"' 3.40 (m) 71.9 3.38 (t, 9.6) 72.4 3.33 (m) 71.2 5 " 3.45 (m) 70.6 3.56 (m) 71.0 3.67 (dd, 9.6, 6.0) 69.3 6 " 1.19 (d, 6.6) 18.1 1.19 (d, 6.0) 17.2 1.25 (d, 6.0) 18.1

¹ H-NMR was measured at 600 MHz, and ¹³ C-NMR at 150 MHz. Also, acetone- d ₆ + D ₂ O with TMS was used as an internal standard.

Example  3. α-Glucosidase Inhibitory Activity of Lutein Derivatives Produced by Gamma Irradiation

To confirm the? -Glucosidase inhibitory activity of the novel lucin derivatives produced by gamma irradiation, an enzyme solution (? -Glucosidase (0.02 unit, 50 μl), substrate (1 mM ρ-nitrophenyl-α-D-glucopyranoside , 50 μl)) was mixed with 50 mM phosphate buffer (pH 7.0) mixed with 5% DMSO (10 μl). After reaction at 37 ° C for 20 minutes, the reaction was stopped by adding 2M NaOH. The amount of ρ-nitrophenyl released using a UV micro-reader was measured at 410 nm. Acarbose was used as a positive control.

As a result, as shown in Table 2, the novel compounds 1 to 3 produced by the irradiation with irradiation showed an increase of the? -Glucosidase inhibitory activity compared to the rutin. In particular, the IC ₅₀ value of the new compound 2 showed the highest inhibitory activity of 11.2 ± 0.7.

compound IC ₅₀ value (μM) Routine > 500 The novel compound 1 23.1 ± 1.2 The new compound 2 11.2 ± 0.7 The novel compound 3 56.2 ± 3.5 Akaboz 310.2 ± 3.6

Claims (6)

A pharmaceutical composition for preventing or treating diabetes mellitus comprising the lutein derivative represented by the following formula (1), (2) or (3) or a pharmaceutically acceptable salt thereof as an active ingredient.
[Chemical Formula 1]

(2)

(3)

The pharmaceutical composition for preventing or treating diabetes according to claim 1, wherein the rutin derivative is produced by irradiating gamma-rays to the rutin. The pharmaceutical composition for preventing or treating diabetes according to claim 2, wherein the gamma ray is irradiated so that an absorbed dose is 20 to 30 kGy. The pharmaceutical composition for preventing or treating diabetes according to claim 1, wherein the rutin derivative has an inhibitory effect on the activity of? -Glucosidase as compared to rutin. A health functional food composition for preventing or ameliorating diabetes comprising an effective ingredient of a lutein derivative represented by the following formula (1), (2) or (3) or a pharmaceutically acceptable salt thereof:
[Chemical Formula 1]

(2)

(3)

The dietary supplement according to claim 5, wherein the health functional food composition is prepared from any one selected from the group consisting of powder, granule, ring, tablet, capsule, candy, syrup and beverage. Health functional food composition.

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