KR101642322B1 - Composition comprising Saussurea grandifolia extract or compound isolated from the same for preventing or treating of diabetic complication - Google Patents

Abstract

The present invention relates to a composition for preventing or treating diabetic complications comprising a Sadler extract or a compound isolated therefrom. The extract of the seeds of the present invention or the compound isolated therefrom of the present invention is excellent in the aldose reductase inhibitory activity and thus can be effectively used for the prevention or treatment of diabetic complications.

Description

A composition for prevention or treatment of diabetic complications comprising a Sadler extract or a compound isolated therefrom, the composition comprising Safflower extract or a compound isolated therefrom,

The present invention relates to a composition for preventing or treating diabetic complications comprising a Sadler extract or a compound isolated therefrom.

Diabetes mellitus is a disease in which blood glucose (blood sugar) concentration in blood is higher than normal, and glucose is released into the urine. It is caused by abnormality of insulin metabolism or abnormal physiological activity of insulin in β-cells of Langerhans islets in the pancreas. Diabetes mellitus causes high osmotic stress to the body's hyperglycemic environment and causes complications such as cataract and kidney disease. Complications due to diabetes vary widely in its type and frequency, including vascular disorders, neurological disorders and infectious diseases.

The pathogenesis of diabetes mellitus has not yet been clarified. However, it is known that the longer the duration of diabetes mellitus and the poorer the blood glucose control, the greater the incidence of diabetes mellitus. Thus, hyperglycemia plays an important role in the development of diabetic complications Has come. Recent DCCT studies in the United States report that hyperglycemia plays an important role in the development of complications by reporting that intensive blood glucose control can significantly reduce the incidence of complications in patients with insulin dependent diabetes mellitus (IDDM) . In addition, there is no complication in patients with long-term uncontrolled diabetes, and DCCT studies have shown that even if the blood glucose control is thorough, the complications can not be completely prevented. As a result, And other genetic factors, which have not yet been clarified, are thought to determine the susceptibility of complications. Recently, studies have been actively conducted on these metabolic disorders. (Giugliano D, Paolisso G, Ceriello A: Oxidative stress and diabetic vascular comlications. Diabetes Care 19: 257-267, 1996)

The mechanism by which hyperglycemia causes complications has not yet been fully elucidated. (1) Chronic hyperglycemia exacerbates the glycation of various proteins in the body, and therefore, (3) the possibility of promoting intracellular osmolality and degeneration by raising the intracellular sorbitol concentration by decreasing myolinositol concentration by the polyol pathway, (3) reducing the intracellular oxidation / reduction ratio (redox status) And (4) the possibility of promoting oxidative damage to the cells. These mechanisms are not mutually exclusive, but rather involve multiple mechanisms or involve different mechanisms as a result of one mechanism And so on.

It has been reported that oxidative stress is induced by lowering the defensive system function against harmful oxygen free radicals occurring at the same time that lipid metabolism abnormality occurs during the production of the final glycation product in hyperglycemic state (Yokozawa, T., et al , 2001, J. of Trad. Med., 18: 107-112). These non-enzymatic glycation reactions and oxidative stress mechanisms are related to each other.

One of the enzymes directly involved in the treatment of complications caused by hyperglycemia is aldose reductase. Aldose reductase is an enzyme involved in the polyol pathway, one of the metabolic pathways in vivo. The polyol pathway consists of two pathways, a pathway in which aldose reductase is involved and reduces glucose to sorbitol, and a pathway in which sorbitol dehydrogenase is involved and sorbitol is dehydrogenated to D-fructose. The polyol pathway is present in the brain, liver, pancreas, kidney, adrenal glands, testes, seminal vesicles, placenta, red blood cells, lens, cornea, iris, retina and peripheral nerves to reduce glucose to sorbitol, To be converted into fructose. However, when the intracellular glucose inflow is increased, glucose produces a large amount of intermediate metabolites and sorbitol through the polyol pathway, and sorbitol is not effectively diffused out of the cell and accumulates in the cell, resulting in increased intracellular osmotic pressure . High osmotic conditions cause variations in the various metabolic processes within the cell and reduce the stability of the cell membrane, resulting in diabetic neuropathy, diabetic nephropathy, diabetic retinopathy, diabetic cataracts (diabetic cataract) and other complications.

Therefore, an aldose reductase inhibitor has been developed and prescribed for the purpose of preventing, alleviating or treating the above-mentioned complications of diabetes, but the prescription is extremely limited due to side effects.

On the other hand, the seedlings are perennial herbaceous plants grown in the deep mountains of each place in Korea. The growth environment is rich in leafiness in the soil and it grows in a semi - The height is 30 ~ 50㎝, the leaves gradually become shorter as they go up, and the petiole length is 5 ~ 12㎝. The surface of the leaf is green, with a slight white light on the backside, with sharp sawtooth on the edge of the leaf. The flower is a purple flower, 4 ~ 6 at the end of the main stem. It blooms in July ~ October, and 4 ~ 6 heads (at the end of the stem, flowers with bloom heads gathered at the end of the stem). The head is made of purple color and has a diameter of 1.8 ~ 2cm, and the corolla corolla is 14 ~ 16.5mm long. The involucre is a bell-shaped, narrow-tipped, white-haired, spider-like. The bracts are arranged in 7 to 10 rows and the inner pieces are 4 to 5 times longer than the outer pieces. The fruit is an achene, it runs in about October, and the white hairs are in the shape of a dagger and the length is about 1㎝. They are distributed in Korea, Japan, and northeastern China. Eat young leaves as herbs. Sudoru is used as a medicinal product for bruises, low back pain, deep sea, genomes, and blood. The main pharmacological ingredients include terpenoids, flavonoids, lignans, sesquiterpene lactones, linolenic acid, chamomile, jacobine, ameleme), and it is known that it has antioxidant, anti-cancer and anti-arteriosclerosis effects. As described above, various pharmacological effects of Suterol are known, but it has not yet been clarified whether the Suterol extract has a preventive or therapeutic effect on diabetic complications.

Therefore, the inventors of the present invention conducted a study on the extracts of the seeds to develop new drugs, and found out that the novel compounds were identified in the seeds extract, and that the extracts of the seeds and the compounds isolated therefrom had excellent aldose reductase inhibitory activity And completed the present invention.

It is an object of the present invention to provide a method of inhibiting the activity of calycopteretin-7-O- alpha -L-rhamnoside or scutellarin-8-C-beta having aldose reductase inhibitory activity -D-glucoside (scutellarein-8-C- [beta] -D-glucoside).

It is also an object of the present invention to provide a composition for preventing or treating diabetic complications comprising an extract of Suratococcus sp. Or a compound isolated therefrom as an active ingredient.

It is an object of the present invention to provide a method of inhibiting the activity of calycopteretin-7-O- alpha -L-rhamnoside or scutellarin-8-C-beta having aldose reductase inhibitory activity -D-glucoside (scutellarein-8-C- [beta] -D-glucoside).

It is also an object of the present invention to provide a composition for preventing or treating diabetic complications comprising an extract of Sodaliae or a compound isolated therefrom as an active ingredient.

The extract of the seeds of the present invention or the compound isolated therefrom of the present invention is excellent in the aldose reductase inhibitory activity and thus can be effectively used for the prevention or treatment of diabetic complications.

The present invention relates to a calicopteretin-7-O- alpha -L-rhamnoside represented by the following formula (I) having an aldose reductase inhibitory activity or a calicopteretin- 8-C-β-D-glucoside represented by the following formula (II).

(I)

[Formula II]

The compounds of the above formulas (I) and (II) are extracted and separated from the seeds.

The compounds of Formulas (I) and (II) above may be prepared as pharmaceutically acceptable salts and solvates according to methods conventional in the art.

Pharmaceutically acceptable salts include acid addition salts formed by free acids. The acid addition salt is prepared by a conventional method, for example, by dissolving the compound in an excess amount of an acid aqueous solution, and precipitating the salt using a water-miscible organic solvent such as methanol, ethanol, acetone or acetonitrile. The same molar amount of the compound and the acid or alcohol (e.g., glycol monomethyl ether) in water may be heated and then the mixture may be evaporated to dryness, or the precipitated salt may be subjected to suction filtration.

As the free acid, an organic acid and an inorganic acid can be used. As the inorganic acid, hydrochloric acid, phosphoric acid, sulfuric acid, nitric acid, tartaric acid and the like can be used. Examples of the organic acid include methanesulfonic acid, p- toluenesulfonic acid, acetic acid, But are not limited to, acetic, citric, maleic, succinic, oxalic, benzoic, tartaric, fumaric, manderic, propionic, lactic, Glucuronic acid, glutamic acid, glutaric acid, glucuronic acid, aspartic acid, ascorbic acid, carbonic acid, vanillic acid, hydroiodic acid and the like can be used.

In addition, pharmaceutically acceptable metal salts can be prepared using bases. The alkali metal or alkaline earth metal salt is obtained, for example, by dissolving the compound in an excess amount of an alkali metal hydroxide or alkaline earth metal hydroxide solution, filtering the non-soluble compound salt, and evaporating and drying the filtrate. At this time, it is preferable for the metal salt to produce sodium, potassium or calcium salt in particular, and the corresponding silver salt is obtained by reacting an alkali metal or alkaline earth metal salt with a suitable silver salt (for example, silver nitrate).

The pharmaceutically acceptable salts of the compounds of formulas (I) and (II) above, unless otherwise indicated, include salts of acidic or basic groups which may be present in the compounds of formula (I) and (II) do. For example, pharmaceutically acceptable salts include the sodium, calcium, and potassium salts of the hydroxy group, and other pharmaceutically acceptable salts of the amino group include hydrobromide, sulfate, hydrogen sulfate, phosphate, hydrogen phosphate, (Salicylate) salts, which are known in the art and which are known in the art, such as, for example, ≪ / RTI >

The present invention also provides a composition for preventing or treating diabetic complications comprising the extract of Sodaliae or a compound represented by the following formula (III), (IV) or (V) isolated therefrom as an active ingredient.

[Formula (III)

In the above formula (III), R ₁ to R ₇ are each independently the same or different and are H, OH, O-Rutinoside, O-Rhamnoside, O-glucoside (O-Glucoside), C-Glucoside, OMe, and C-xyloside.

Preferably, in the above formula (III)

R ₁ is H, OH, O-Rutinoside, O-Rhamnoside or O-Glucoside;

R ₂ is OH or O-Glucoside,

R ₃ is H, OH, C-Glucoside or C-xyloside;

R ₄ is H, OH, O-Rhamnoside or OMe;

R ₅ is H, OH or C-Glucoside,

R < ₆ > is H or OH;

R ₇ is OH.

[Formula IV]

[Formula V]

As used herein, the term "extract" means an active ingredient isolated from a natural product, i.e., a substance exhibiting the desired activity. The extract can be obtained by an extraction process using water, an organic solvent, or a mixed solvent thereof, and includes all the forms that are formulated by using the dry powder of the extract. In addition, the above-mentioned extract also includes a fraction obtained by fractionating the extract obtained through the above extraction process.

The extract of the seeds of the present invention or the compound isolated therefrom can be obtained by the following process.

The seedlings were collected from Korea 's Ulleungdo Island, and the specimens were certified at the Korean National Arboretum. The extraction site of the seed coat is not limited thereto, but the leaves and the stem of the seed coat are preferable. The sucrose is extracted with water, an alcohol having 1 to 4 carbon atoms, or a mixed solvent thereof, and the extract is concentrated under reduced pressure. The alcohol having 1 to 4 carbon atoms is a lower alcohol selected from the group consisting of methanol, ethanol, propanol and isopropanol, more preferably methanol. The concentrate is suspended in distilled water and successively fractionated with hexane, methylene chloride, ethyl acetate and butanol to obtain respective solvent fractions. The aldose reductase inhibitory activity of the sucrose ethyl acetate fraction in the sucrose fraction was highest. Thus, chromatographies (paper, TLC, column, etc.) were carried out from the fractions of the seeding ethyl acetate using silica or paper to isolate eleven compounds as the active compounds of the formula (III), (IV) and (V) do. That is, the isolated compounds include vitexin (1), genkwanin-5-O-glucoside (2), calicheptoretin-7-O- L-laminoside (3), vitexin 2 "-p-hydroxybenzoate (4), kaempferol-3-O-rutinoside Isovitexin (6), quercitrin (7), isoquercitrin (8), 1,5-di-O-caffeoylquinic acid (1,5) -di-O-caffeoylquinic acid (9), vicenin 1 (10) and scutellarane-8-C-? -D-glucoside (11).

The chromatography may be thin layer chromatography (TLC) or column chromatography using silica as a carrier. The TLC is a method in which a silica gel is thinly coated on a support such as a glass plate or a plastic, and the extract is dripped on the support, followed by development with a developing solvent. The column chromatography is a method in which silica is packed in a column, the extract is loaded on the column, and developed with a developing solvent. For mass production of the extract, column chromatography is preferably used.

The extract of the present invention or the compound isolated therefrom is excellent in the inhibitory activity of aldose reductase (AR), and thus can be effectively used for the prevention or treatment of diabetic complications.

Such diabetic complications include diabetic neuropathy, diabetic nephropathy, diabetic myocardial infarction, diabetic retinopathy, diabetic cataract, and diabetic neuropathy. Diabetic foot, and the like, but are not limited thereto.

In addition, the composition of the present invention may contain one or more known active ingredients having the effect of preventing or treating diabetic complications, together with the Sodalaceae extract or the compound isolated therefrom.

The pharmaceutical compositions of the present invention may further comprise suitable carriers, excipients and diluents conventionally used in the manufacture of pharmaceutical compositions. In addition, it can be formulated into oral dosage forms such as powders, granules, tablets, capsules, suspensions, emulsions, syrups and aerosols, sterilized injection solutions, suppositories and transdermal dosage forms according to a conventional method. Examples of carriers, excipients and diluents that can be included in the composition include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methylcellulose, Microcrystalline cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil. If necessary, it is formulated using a diluent such as a filler, an extender, a binder, a wetting agent, a disintegrant, a surfactant, or an excipient.

When the pharmaceutical composition of the present invention is formulated into a solid preparation for oral administration, solid preparations for oral administration include tablets, pills, powders, granules, capsules and the like. These solid preparations may contain at least one excipient For example, it is formulated by mixing starch, calcium carbonate, sucrose or lactose, gelatin and the like. In addition to simple excipients, lubricants such as magnesium stearate and talc may also be used. In addition, the pharmaceutical composition of the present invention may be formulated into a liquid preparation for oral administration. Liquid preparations for oral administration include suspensions, solutions, emulsions and syrups. These liquid preparations may contain various excipients in addition to commonly used inert diluents (for example, purified water, ethanol, liquid paraffin) For example, wetting agents, sweeteners, fragrances, preservatives and the like may be included. In addition, the pharmaceutical composition of the present invention may be formulated into preparations for parenteral administration, preferably intraperitoneal administration. Formulations for parenteral administration include sterilized aqueous solutions, non-aqueous solutions, suspensions, emulsions, freeze-dried preparations, and suppositories. As the sterilized aqueous solution, a suitable buffer solution such as Hank's solution, Ringer's solution or physically buffered saline can be used. As the non-aqueous solvent, propylene glycol, polyethylene glycol, olive oil, The same vegetable oil, an injectable ester such as ethyl oleate, or the like can be used. If desired, preservatives, stabilizers, wetting or emulsifying agents, salts for controlling osmotic pressure and / or buffers may be used. On the other hand, in the case of suppositories, witepsol, macrogol, tween 61, cacao paper, laurin, glyceroglatin and the like may be used.

The term "administering" as used herein is meant to provide any desired composition of the invention to a subject in any suitable manner. The composition formulated in the above manner may be administered through various routes including parenteral or oral (transdermal, subcutaneous, intravenous, muscular, abdominal, etc.) in an effective amount. The "effective amount" as used herein refers to an amount that shows a preventive or therapeutic effect when administered to a patient. The dose of the composition according to the present invention can be appropriately selected depending on the route of administration, subject to be administered, age, sex, weight, individual difference, and disease state. Preferably, the composition of the present invention may contain the active ingredient in an amount of 0.1 to 10000 mg / kg body weight / day, more preferably 10 to 1000 mg / kg body weight / day, Can be administered in an effective amount. The composition of the present invention can be used alone or in combination with methods using surgery, radiation therapy, hormone therapy, chemotherapy, and biological response modifiers for the prevention and treatment of inflammatory diseases.

The composition of the present invention may be added to a health functional food for the purpose of preventing or improving diabetic complications. When the seed extract of the present invention or a compound isolated therefrom is used as a food additive, the seed extract or the compound isolated therefrom can be used as it is or can be used together with other food or food ingredients, Lt; / RTI > The amount of the active ingredient to be mixed can be suitably determined according to the intended use (prevention, health or therapeutic treatment). Generally, the seedling extract of the present invention or the compound isolated therefrom is added in an amount of not more than 15% by weight, preferably not more than 10% by weight based on the raw material. However, in the case of long-term intake for the purpose of health and hygiene or for the purpose of controlling 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 food. Examples of foods to which the above substances can be added include dairy products including meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen and other noodles, gums, ice cream, various soups, drinks, tea, Alcoholic beverages, and vitamin complexes, all of which include health foods in a conventional sense.

The health beverage composition of the present invention may contain various flavors or natural carbohydrates as an additional ingredient such as ordinary beverages. The natural carbohydrates may be monosaccharides such as glucose and fructose, disaccharides such as maltose and sucrose, natural sweeteners such as dextrin and cyclodextrin, synthetic sweeteners such as saccharine and aspartame, and the like. The ratio of the natural carbohydrate is generally about 0.01 to 10 g, preferably about 0.01 to 0.1 g per 100 ml of the composition of the present invention.

In addition to the above, the composition of the present invention may further contain various nutrients, vitamins, electrolytes, flavors, colorants, pectic acid and salts thereof, alginic acid and salts thereof, organic acids, protective colloid thickeners, pH adjusters, stabilizers, preservatives, glycerin, A carbonating agent used in a carbonated beverage, and the like. In addition, the composition of the present invention may comprise flesh for the production of natural fruit juices, fruit juice drinks and vegetable drinks. These components may be used independently or in combination. Although the ratio of such additives is not critical, it is generally selected in the range of 0.01 to 0.1 parts by weight per 100 parts by weight of the composition of the present invention.

Hereinafter, preferred embodiments and production examples are shown to facilitate understanding of the present invention. However, the following Examples and Preparation Examples are provided only for the purpose of easier understanding of the present invention, and the present invention is not limited by the Examples and Production Examples.

EXAMPLES Example 1: Extract of Sodalaceae and isolation of active ingredient therefrom

1-1. Preparation of extracts and fractions of seedlings

(2099.8 g) was shaken and systematically extracted using methanol (7 L). The extract was extracted 5 times and then depressurized and concentrated with a vacuum concentrator (CCA-1100, EYELA) at 65 to 75 ° C, The extract (271.0 g) was obtained. The suspo methanol extract was suspended in distilled water and successively fractionated with hexane, methylene chloride, ethyl acetate and butanol to obtain each of the desolate solvent fractions. The fraction of the distillation product (41.3 g), the sucrose methylene chloride (MC ) Fractions (8.8 g), the seeding ethyl acetate (EtOAc) fraction (8.1 g) and the sucrose butanol (BuOH) fraction (21.9 g).

1-2. Separation of the active ingredient from the sucrose fraction

The ethyl acetate fraction having the highest inhibitory effect on aldose reductase was separated from the seedling fractions obtained in 1-1 above by using silica gel column chromatography into 10 flavonoids and 1 phenylpropanoid . The results of the determination of the active components by spectral analysis ( ¹ H- and ¹³ C-NMR, and MS) of these compounds are shown in Table 1.

compound R ₁ R ₂ R ₃ R ₄ R ₅ R ₆ R ₇ One H OH H OH C-Glucoside H OH 2 H O-Glucoside H OMe H H OH 3 OH OH OH O-Rhamnoside OH H OH 5 O-Rutinoside OH H OH H H OH 6 H OH C-Glucoside OH H H OH 7 O-Rhamnoside OH H OH H OH OH 8 O-Glucoside OH H OH H OH OH 10 H OH C-xyloside H C-Glucoside H OH 11 H OH OH OH C-Glucoside H OH Compound 4 Compound 9

As shown in Table 1, vitexin (1), genkwanin-5-O-glucoside (2), bitexin 2 "-p-hydroxybenzoate vitexin 2 "-p-hydroxybenzoate (4), kaempferol-3-O-rutinoside (5), isovitexin (6), quercitrin (7), isoquercitrin (8), 1,5-di-O-caffeoylquinic acid (9), and vicenin 1 ) (10) was identified as a known compound.

Among these compounds, the compounds (3) and (11) were identified as the compounds of the formulas (I) and (II) as novel compounds and their spectral analyzes ( ¹ H- and ¹³ C- Are shown in Table 2.

Compound 3 Compound 11 隆_H (J in Hz) δ _C HMBC 隆_H (J in Hz) δ _C HMBC 2 156.0 163.9 3 136.2 6.63 (s) 105.8 4 175.9 180.2 5 161.9 164.0 6 132.5 131.5 7 154.9 162.0 8 127.4 110.0 9 155.8 158.1 10 101.0 108.0 One' 120.4 120.2 2' 7.71 (d, 8.5) 129.9 C-4 ' 7.96 (br d) 129.4 3 ' 6.89 (d, 8.5) 114.5 C-1 ' 6.87 (d, 8.5) 115.1 C-1 ' 4' - 159.4 - 160.9 5 ' 6.89 (d, 8.5) 114.5 C-1 ' 6.87 (d, 8.5) 115.1 C-1 ' 6 ' 7.71 (d, 8.5) 129.9 C-4 ' 7.96 (br d) 129.4 5'-OH 12.58 (s) 13.63 (s) Rha-1 5.28 (br d) 100.3 C-7 Rha-2 4.92 68.4 Rha-3 3.97 70.6 Rha-4 3.30 71.0 Rha-5 3.17 69.0 Rha-6 0.79 (d, 6.0) 17.1 Rha-5 Glc-1 4.50 (d, 8.0) 76.0 C-9 Glc-2 3.30 72.4 Glc-3 3.38 79.0 Glc-4 3.17 68.5 Glc-5 3.51 81.0 Glc-6 3.72, 3.86 62.1

 The compound (3) was an amorphous nor-red powder and was identified as calycopteretin-7-O- alpha -L-rhamnoside. The structural formula was identified by the formula (I) Respectively.

(I)

The FAB MS results showed a positive molecular ion peak [M + H] ⁺ at m / Z 465. As a result of IR spectrum analysis, α, β-unsaturated carbonyl (1,656 cm ^-1 ) (1,450 cm ^-1 ) and a hydroxyl group (3,375 cm ^-1 ). The UV spectra in methanol showed maximum values at 265, 281 and 337 nm.

The compound (11) was identified as a yellow rubber, scutellarein-8-C-? -D-glucoside and represented by the formula (II).

[Formula II]

The result of FAB MS showed a negative molecular ion peak [M - H] ^- at m / Z 447. As a result of IR spectrum analysis, the hydroxyl group (3,365 cm ^-1 ) and the α, β-unsaturated carbonyl ^-1 ). Also, the UV spectrum in methanol showed maximum values at 268, 293 and 394 nm.

Experimental Example 1. Measurement of Aldose Reductase Inhibitory Effect of Extracts and Fractions of Seuldogae

In order to confirm the aldose reductase inhibitory effect of the seed extract of the present invention, the following experiment was conducted.

Specifically, the lens of rat (Rat, 250-280 g) was extracted and homogenized by adding 0.5 ml of sodium buffer per lens. This was centrifuged at 10,000 rpm for 20 minutes at 4 ° C, and the supernatant was taken as an enzyme source. The enzyme source prepared above was reacted with 0.025 M DL-glyceraldehyde as a substrate and the decrease rate of NADPH absorbance at 340 nm was measured using a spectrophotometer (Spectra). And 3-tetramethylene glutaric acid (TMG), a strong aldose reductase inhibitor, was used as a positive control. As a negative control, DMSO was used.

Table 3 shows the experimental results on the inhibitory effect of aldose reductase on the extracts and fractions of seedlings.

density
(μg / mL) Inhibition rate
(%) IC ₅₀
(μg / mL)
MeOH extract
5 85.19
0.84
One 56.64 0.5 37.16 n-Hexane fraction 10 21.58 - MC fraction 10 46.48 -
The EtOAc fractions
One 93.41
0.12
0.1 57.21 0.05 24.37
The n-BuOH fraction
10 84.07
3.33
5 67.28 One 8.67
TMG
5 78.48
0.96
One 58.96 0.5 32.16

As shown in Table 3, it was confirmed that the IC ₅₀ of the seedlings methanol extract was 0.84 μg / mL, and thus the aldose reductase inhibitory effect was excellent. In addition, it was confirmed that the IC ₅₀ value of the sucrose hexane fraction, the methylene chloride fraction, the ethyl acetate fraction and the butanol fraction was 0.12 μg / mL, and the aldose reductase inhibitory activity was the highest.

Experimental Example 2. Measurement of the inhibitory effect of the compound 1-11 isolated from the extract of Sodalaceae on aldose reductase

The aldose reductase inhibitory effect of the compounds 1 to 11 isolated from the seeds extract was measured. That is, in order to measure the effect of inhibiting the lens Aldose reductase in rats, the percent inhibition and the IC ₅₀ value were measured. In particular, compounds exhibiting inhibitory effects of greater than 50% measured the aldose reductase inhibitory effect at three concentrations. The measurement was performed in the same manner as in Experimental Example 1 and compared using TMG as a positive control. The measurement results are shown in Table 4.

compound density
(μg / mL) Inhibition rate
(%) IC ₅₀
(μg / mL)
One
10 96.0
1.45
5 80.8 0.1 21.1
2
10 74.8
12.02
5 51.2 One 23.5
3
10 81.7
3.81
5 70.0 0.5 36.7 4

10 60.1
14.66
5 34.3 One 8.1 5 10 21.6 -
6
10 50.5
22.83
5 31.2 One 15.7
7
10 97.7
0.16
One 69.5 0.1 45.7
8
10 87.2
0.32
One 65.9 0.1 46.9
9 10 95.4
0.08
0.1 64.6 0.05 45.7 10 10 24.9 - 11 10 17.3 -
TMG
5 78.5
5.16
One 59.0 0.5 32.2

As shown in Table 4, flavonoids and phenylpropanoids 10 except compounds (5), (10) and (11) showed excellent aldose reductase inhibitory activity. L-laminocide (3), quercitrin (7), isoquercitrin (2), calichectotretin-7-O- alpha -L- 1isoquercitrin (8) and 1,5-di-O-caffeoylquinic acid (9) all showed 70% or more inhibitory effect on aldose reductase. Among them, quercitrin (7) and isoquercitrin (8) showed a high inhibitory activity (IC ₅₀ , 0.16 and 0.32 μM, respectively) at 1.5 times of TMG and 1,5-di- (9) showed the highest aldose reductase inhibitory activity (IC ₅₀ , 0.08 μM) over 6.5 times that of TMG.

Formulation Example 1. Pharmaceutical preparation

1-1. Manufacture of Powder

20 mg < RTI ID = 0.0 >

Lactose 100 mg

Talc 10 mg

The above components are mixed and filled in airtight bags to prepare powders.

1-2. Manufacture of tablets

Surfactant extract or compounds isolated therefrom 10 mg

Corn starch 100 mg

Lactose 100 mg

Magnesium stearate 2 mg

After mixing the above components, tablets are prepared by tableting according to the usual preparation method of tablets.

1-3. Preparation of capsules

Surfactant extract or compounds isolated therefrom 10 mg

Crystalline cellulose 3 mg

Lactose 14.8 mg

Magnesium stearate 0.2 mg

The above components are mixed according to a conventional capsule preparation method and filled in gelatin capsules to prepare capsules.

1-4. Injection preparation

Surfactant extract or compounds isolated therefrom 10 mg

180 mg mannitol

Sterile sterilized water for injection 2974 mg

Na ₂ HPO ₄ .2H ₂ O 26 mg

(2 ml) per 1 ampoule in accordance with the usual injection preparation method.

1-5. Manufacture of liquid agent

20 mg < RTI ID = 0.0 >

10 g per isomer

5 g mannitol

Purified water quantity

Each component was added and dissolved in purified water according to the usual liquid preparation method, and the lemon flavor was added in an appropriate amount. Then, the above components were mixed, and purified water was added thereto. The whole was added with purified water to adjust the total volume to 100 ml, And sterilized to prepare a liquid preparation.

Formulation Example 2. Food preparation

2-1. Manufacture of health food

Surfactant extract or a compound isolated therefrom 100 mg

Vitamin mixture quantity

Vitamin A acetate 70 μg

Vitamin E 1.0 mg

Vitamin B1 0.13 mg

0.15 mg of vitamin B2

Vitamin B6 0.5 mg

Vitamin B12 0.2 μg

Vitamin C 10 mg

Biotin 10 μg

Nicotinic acid amide 1.7 mg

Folic acid 50 μg

Calcium pantothenate 0.5 mg

Mineral mixture quantity

1.75 mg of ferrous sulfate

0.82 mg of zinc oxide

Magnesium carbonate 25.3 mg

Potassium monophosphate 15 mg

Secondary calcium phosphate 55 mg

Potassium citrate 90 mg

Calcium carbonate 100 mg

Magnesium chloride 24.8 mg

Although the composition ratio of the above-mentioned vitamin and mineral mixture is comparatively mixed with a composition suitable for health food as a preferred embodiment, the compounding ratio may be arbitrarily modified, and the above ingredients are mixed according to a conventional method for producing healthy foods , Granules can be prepared and used in the manufacture of health food compositions according to conventional methods.

2-2. Manufacture of health drinks

Surfactant extract or a compound isolated therefrom 100 mg

Vitamin C 15 g

Vitamin E (powder) 100 g

19.75 g of ferrous lactate

3.5 g of zinc oxide

Nicotinic acid amide 3.5 g

Vitamin A 0.2 g

Vitamin B1 0.25 g

Vitamin B2 0.3 g

Water quantification

The above components were mixed according to a conventional health drink manufacturing method, and the mixture was stirred and heated at 85 DEG C for about 1 hour. The solution thus prepared was filtered and sterilized in a sterilized 2 liter container, It is used in the production of the health beverage composition of the invention.

Although the compositional ratio is relatively mixed with a component suitable for a favorite drink, it is also possible to arbitrarily modify the compounding ratio according to the regional or national preference such as the demand class, the demanding country, and the use purpose.

Claims (24)

delete delete delete delete delete delete delete delete delete delete A pharmaceutical composition for preventing or treating diabetic complications, which comprises a compound represented by the following formula (V) isolated from a seeds extract:
[Formula V]

delete 12. The method of claim 11, wherein the compound of formula (V) is 1,5-di-O-caffeoylquinic acid. A pharmaceutical composition. delete delete 12. The pharmaceutical composition for preventing or treating diabetic complications according to claim 11, wherein the extract of Sodolec is extracted with water, an alcohol having 1 to 4 carbon atoms or a mixed solvent thereof. The pharmaceutical composition according to claim 16, wherein the alcohol having 1 to 4 carbon atoms is methanol, ethanol, propanol or isopropanol. 12. The method according to claim 11, wherein the extract of the seeds is extracted with water, an alcohol having 1 to 4 carbon atoms or a mixed solvent thereof, distilled water is added thereto, and then hexane, methylene chloride (MC), ethyl acetate EtOAc) and butanol (BuOH). The pharmaceutical composition for preventing or treating diabetic complications according to claim 1, 12. The pharmaceutical composition according to claim 11, wherein the extract is a fragrance of sucrose ethyl acetate. 12. The method according to claim 11, wherein the diabetic complication is at least one selected from the group consisting of diabetic neuropathy, diabetic nephropathy, diabetic myocardial infarction, diabetic retinopathy, diabetic cataract, and diabetic foot ulcer. A pharmaceutical composition for the prevention or treatment of complications. A food composition for preventing or ameliorating diabetic complications comprising a compound represented by the following formula (V) isolated from a seedling extract:
[Formula V]

delete 22. The food composition according to claim 21, wherein the extract of Sodolecum is extracted with water, an alcohol having 1 to 4 carbon atoms or a mixed solvent thereof. delete