KR100867370B1 - Pharmaceutical compositions for the prevention and treatment of diabetic complications or obesity containing Sophora flavascens extracts or the isolated 5-methylsophoraflavanone B therefrom as an active ingredient - Google Patents

Abstract

The present invention relates to a pharmaceutical composition for preventing and treating diabetic complications or obesity, which contains a ginseng extract or 5-methylsophoraflavanone B isolated therefrom as an active ingredient. -Methylsofolaflavanone B exhibits the inhibitory effect of aldose reductase which causes diabetic complications, and also prevents obesity by inhibiting adipocyte differentiation and thus the composition according to the present invention prevents or treats diabetic complications or obesity. It can be useful to

Ginseng, Methylsofolaflavanone B, Diabetes Complications, Obesity, Prevention and Treatment

Description

Pharmaceutical compositions for the prevention and treatment of diabetic complications or obesity containing Sophora flavascens extracts or the isolated 5-methylsophoraflavanone B therefrom as an active ingredient}

Figure 1 shows the stained cell line appearance photograph of the normal differentiated adipocytes (a) and adipocytes (b) treated with 5-methylsophoraflavanone B,

Figure 2 shows the effect of inhibiting adipocyte differentiation by 5-methylsophoraflavanone B of the present invention,

Figure 3 shows the results of microscopic observation of normally differentiated adipocytes (a) and adipocytes (b) treated with 5-methylsophoraflavanone B.

The present invention relates to a pharmaceutical composition for the prevention and treatment of diabetic complications or obesity, containing a ginseng extract or 5-methylsophoraflavanone B isolated therefrom as an active ingredient.

Diabetes is a chronic disease in which glucose is released into the urine due to a lack of insulin, which leads to higher than normal levels of glucose in the blood. There are many diseases associated with diabetes that may be present in almost all diseases other than diabetes. However, the incidence of complications is high and important include retinopathy, neuropathy and nephropathy.

As diabetes increases the body's blood sugar levels for a long time, chronic complications occur as glycation products invade the kidneys, nerves, retinas, and large and small blood vessels throughout the body. Over half of all diabetic complications are diabetic retinopathy and adult blindness. It is the biggest cause of onset. Symptoms include vision problems, glaucoma, and vision loss.

In addition, diabetic neuropathy causes peripheral neuropathy due to abnormal carbohydrate and lipid metabolism. Symptoms include decreased keyboard loss, loss, perception disorders, and limb pain. In particular, the lower extremity may complain of symptoms such as nighttime muscle pain, and the burning of the soles of the feet, hyperalgesia and tactile blunts. Usually it is symmetrical and movement disorder is rare. In addition, autonomic neuropathy, left and right swelling of the pupil, sweating abnormalities, sores (陰 蔿), residual urination, delayed urination, urinary tract (폐), constipation and diarrhea can be seen.

Nephropathy is also very common and is considered the most important cause of death of complications of diabetes. Diabetics have been shown to have a 20-fold higher risk of renal failure than normal patients. Symptoms include glomerulosclerosis, proteinuria, and kidney failure. Nephropathy has been reported to be caused by excessive aldose reductase or oxidative stress.

Mechanisms leading to diabetic complications include osmotic stress caused by changes in the mechanism of non-enzymatic glycation of protein and polyol pathway and oxidative stress caused by free radicals. ) And the like.

In non-enzymatic glycosylation of proteins, amino acid groups, such as lysine residues, are condensed and cross-reacted with reducing sugars without enzymatic action to produce advanced glycation endproducts (AGEs). Final glycosylation products are irreversible reaction products, so once they are produced they do not degrade when blood sugar returns to normal, but accumulate in tissues during protein survival, resulting in abnormal changes in tissue structure and function (Vinson, JA et al., J. Nutritinal Biochemistry, 7, 559-663, 1996; Smith, PR et al., Eur. J Biochem., 210, 729-739, 1992). Non-enzymatic protein glycosylation results in glycation of proteins such as basement membrane, plasma albumin, lens protein, fibrin, and collagen, and the resulting glycosylated product abnormally alters the structure and function of tissues, resulting in diabetic retinopathy, It causes chronic diabetic complications such as cataract, diabetic nephropathy and diabetic neuropathy.

In addition, it has been reported that abnormal glycation metabolism occurs in the process of producing the final glycosylated product in hyperglycemic state and oxidative stress is caused by deterioration of defense system function against harmful oxygen free radicals (Yokozawa, T. et al. , J of Trad. Med., 18, 107-112, 2001). As such, the mechanism of action of non-enzymatic glycosylation and oxidative stress is related.

The polyol refers to an alcohol reduced by aldose reductase (AR) from aldose or ketose. In general, when glucose enters the cell, hexokinase converts it into 6-glucose and then breaks down the pathway. However, when the concentration of glucose increases, hexokinase that binds to it is saturated, and excess glucose binds to aldose reductase, which is less affinity than hexokinase, and is metabolized to sorbitol to accumulate sorbitol in cells. Aldose reductase is present in the retina, sclera, lens, kidney, erythrocytes, brain, muscle, liver and peripheral nerves, especially in the peripheral nerves can be a problem because glucose permeation is not affected by insulin. In other words, when hyperglycemia causes sorbitol to accumulate in Schumann cells, osmotic pressure may increase, resulting in edema, necrosis, and collapse of myelin sheath, and water may enter due to continuously increased osmotic pressure, leading to complications of diabetes ( Diabetes, Eung-Jin Kim et al., Korean Diabetes Association, Korea Medicine, p. 483; Soulis-Liparota, T. et al., Diabetologia, 38, 357-394, 1995).

Final glycation end products have been reported to activate aldose reductase (AR), the main enzyme of the polyol pathway, in human microvascular endothelial cells (Nakamura, N. et al., Free Radic Biol. Med., 29, 17-25, 2000). In a steady state, aldose reductase has a very low affinity for glucose, but at high concentrations of glucose, aldose reductase is activated to excessively reduce glucose to sorbitol, and this sorbitol is converted to fructose by sorbitol dehydrogenase. This fructose is about 10 times faster than non-enzymatic glycosylation of protein. Thus, high concentrations of fructose bind to the protein and eventually accelerate the formation of the final glycated product.

Renal complications are very common and are considered to be the most important cause of death of diabetes complications. In addition, diabetics have been found to have a 20-fold higher risk of renal failure than normal patients. AS-3201 ((R) -2- (4-bromo-2-fluorobenzyl) -1,2,3,4-tetrahydropyrrolo [1,2-], an aldose reductase inhibitor, for the treatment of kidney complications a] pyrazine-4-spiro-3'-pyrrolidine-1,2 ', 3,5'-tetrone), tollestat, fornalstat and epilestat However, research on their therapeutic effects is still ongoing. In addition, as part of the development of a therapeutic agent for diabetic complications, aldose reductase inhibitory activity has been shown, and at the same time, studies on natural substances having anti-diabetic, antioxidant and hypoglycemic effects have been actively conducted.

Once diabetes develops and blood sugar does not drop quickly, diabetic complications should be taken to prevent diabetic complications. Currently, aminoguanidine HCl, the only synthetic agent for protein glycosylation inhibitors, is a nucleophilic hydrazine, which binds to the products of condensation reactions and prevents cross-linking with proteins, thereby inhibiting the formation of the final glycation product as a complication. Delay or prevent progression (Brownlee, M. et al., Sciences, 232, 1629-1632, 1986; Edelstein, D. et al., Diabetes, 41, 26-29, 1992). Aminoguanidine HCl is the most promising synthetic drug for the prevention and treatment of diabetic complications, but it has been progressed up to Phase 3 clinical trials, but there is a problem that toxicity is induced upon prolonged administration.

A study by the UK's Diabetes Control and Complication Trial Research Group found that chronic insulin complications of type 2 (insulin-independent) diabetics, who account for the majority of diabetes, are controlled by strong insulin therapy. Although clinical studies report that it slows down the incidence and duration of the onset, complications develop in most diabetic patients after 5 to 10 years.

Mechanisms for inhibiting diabetic complications include the polyol pathway, the advanced glycation end-products (AGEs) pathway, the hexosamine pathway, and the protein kinase C pathway. (Nature, 414, p813, 2001), and blocking the mechanisms involved is completely independent of the mechanisms for developing therapeutic drugs for diabetes.

In other words, the drug developed through the mechanism for developing diabetes treatment can not be said to have an effect on diabetic complications, and the drugs effective for diabetic complications are not effective at all, such as blood sugar or insulin resistance. For example, epilestat (ONO-2235, an aldose reductase inhibitor), a diabetic complication inhibitor sold in Japan, has no effect on blood glucose lowering or insulin resistance in experimental animals and clinical trials. It is known to treat diabetic complications by inhibiting DOS reductase and final glycation products. Therefore, the effect on diabetes and the effect on diabetic complications can be said to be a separate problem.

In addition, obesity, which is directly or indirectly related to diabetes and is one of the chronic diseases of modern people, has emerged as one of the most serious health problems in the world due to changes in eating habits and lifestyles caused by industrialization and rising income levels. It was defined as a disease by the Organization (WHO). In addition, obesity is known to be directly or indirectly related to adult diseases such as diabetes, hypertension, hyperlipidemia, heart disease and various cancers. Known mechanisms of action of obesity treatments include appetite suppression, tropical death promotion, digestion suppression, and hormone regulation. Distinctly, inhibiting the process of differentiating progenitor cells into adipocytes can regulate the number of adipocytes in the body, and thus can be used to treat obesity based on this mechanism of action. Differentiation into adipocytes is regulated by signaling between intracellular regulatory factors such as Pref-1, Foxo1, Foxa2, SMAD-3, Wnt-10b, GATA-2, GATA-3, and PPAR. Therefore, by screening for drugs that may affect these intracellular signaling agents, therapeutic agents for obesity can be identified (Harp., Curr. Opin. Lipidol., 15, 303-307, 2003; Tong, et al. , Science, 290).

On the other hand, it has been reported that the compound isolated from Sophora flavescens has anti-diabetic effect by inhibiting alpha-glucosidase and beta-amylase (Jin Hyo Kim et al., Biol) Pharm Bull., 29 (2), 302-305, 2006). Flavonoids derived from the ginseng of the above literature include kushenol A, kurarinone, sophoraflavanone G, 2'-methoxykurarinone, kurarinol , 8-prenylkaempferol, isoxanthohumol, kuraridin, maackian and the like are known. However, the effects of these flavonoids on the treatment and prevention of diabetes complications have not been revealed.

Accordingly, the present inventors found for the first time in the world that 5-methylsophoraflavanone B, a single compound isolated from red ginseng, not only inhibits the activity of aldose reductase, but also inhibits the adipocyte differentiation process. The present invention was completed by finding out that obesity or a disease directly or indirectly related to obesity can be prevented and treated.

It is an object of the present invention to provide a pharmaceutical composition for preventing and treating diabetic complications comprising Gosam extract or 5-methylsophoraflavanone B isolated therefrom as an active ingredient.

Another object of the present invention to provide a pharmaceutical composition for the prevention and treatment of obesity containing a ginseng extract or 5-methylsophoraflavanone B isolated therefrom as an active ingredient.

Another object of the present invention to provide a health food for preventing and improving diabetic complications containing a ginseng extract or 5-methylsophoraflavanone B isolated therefrom as an active ingredient.

In order to achieve the above object, the present invention provides a pharmaceutical composition for the prevention and treatment of diabetes complications or obesity containing a ginseng extract or 5-methylsophoraflavanone B isolated therefrom as an active ingredient.

 In another aspect, the present invention provides a health food for preventing and improving diabetic complications containing a ginseng extract or 5-methylsophoraflavanone B isolated therefrom as an active ingredient.

Hereinafter, the present invention will be described in detail.

The present invention provides a pharmaceutical composition for preventing and treating diabetic complications containing the extract of ginseng as an active ingredient.

The ginseng extract is preferably obtained by extracting from the root of the ginseng, the ginseng can be used without limitation, such as those grown or commercially available. The method of preparing the ginseng extract may use a conventional extraction method in the art. The dried ginseng extract is extracted with ethanol to obtain an extract, and then the extract is filtered and concentrated under reduced pressure to obtain an ethanol extract of red ginseng.

The present invention also provides a pharmaceutical composition for preventing and treating diabetic complications containing 5-methylsophoraflavanone B represented by the following Chemical Formula 1 as an active ingredient, isolated from a ginseng extract.

The compound 5-methylsophoraflavanone B is extracted and separated from red ginseng.

The compound according to the present invention is a representative active ingredient present in red ginseng, a method for separating the compound may be carried out as follows.

The present invention extracts the ginseng root dry matter with C ₁ ~ C ₄ alcohol to obtain a ginseng extract (step 1); And a method of preparing 5-methylsophoraflavanone B separated from the extract of red ginseng, which comprises the step of performing high-performance liquid chromatography on the extract obtained in step 1 to separate and purify the compound (step 2). to provide.

Step 1 according to the present invention is a step of obtaining a ginseng extract by extracting the dried ginseng root dried with alcohol of C ₁ ~ C ₄ . The alcohol may be a lower alcohol such as methanol, ethanol, propanol, butanol and the like, and preferably ethanol.

In step 1 according to the present invention, the method for obtaining the alcohol extract is pulverized dried alpine roots into an appropriate size and put into an extraction container to add an appropriate amount of ethanol to extract at room temperature to obtain an extract. The process may be repeated several times or more in order to increase the extraction efficiency of the ginseng. The extract is filtered and concentrated under reduced pressure to prepare an ethanol extract of ginseng.

Step 2 according to the present invention is a step of separating and purifying 5-methylsophoraflavanone B by performing high performance liquid chromatography on the extract obtained in step 1 above.

The ethanol extract may be separated into 5-methylsophoraflavone B by performing high performance liquid chromatography, wherein 3,7,4'-trihydroxy-5-methoxy-8-prenylflavanone and cusenol N, curarinone, 2'-methoxycurarinone, sophoraflavanone G, lycanone A and curaridine can also be isolated. The chromatography may be performed using a mixture of water and acetonitrile gradient gradient from 35 vol% acetonitrile to 70 vol% acetonitrile as the developing solvent, wherein the dissolution rate is 5 to 15 ml per minute, run time Silver is preferably for 0.5 to 2 hours.

Furthermore, the present invention provides an inhibitor of aldose reductase activity containing Gosam extract or 5-methylsophoraflavanone B isolated therefrom as an active ingredient.

In order to confirm the aldose reductase inhibitory activity of the ginseng extract according to the present invention through the experiment on the enzyme precipitate of the lens of the rat, the ginseng extract of the present invention inhibits the activity of aldose reductase by 63% Could know. In addition, not only 5-methylsophoraflavanone B isolated from the ethanol extract of Ginseng according to the present invention, but also 3,7,4'-trihydroxy-5-methoxy-8-prenylflavanone separated together, In the experiments conducted to confirm the aldose reductase inhibitory activity of cusenol N, curarinone, 2'-methoxycurarinone, sophoraflavanone G, lycanone A and curardine, all compounds were subjected to aldose reductase. It was found that there is an inhibitory activity, and particularly 5-methylsophoraflavone B of the present invention showed the highest inhibitory activity (see Table 2 ).

The mechanisms that lead to diabetic complications are largely explained by non-enzymatic glycosylation of proteins, osmotic stress caused by changes in the mechanism of the polyol pathway, and oxidative stress caused by free radicals, wherein the polyol is aldose from aldose or ketose. It means alcohol reduced by reductase. In general, when glucose enters the cell, hexokinase converts it into 6-glucose and then breaks down the pathway. However, when the concentration of glucose increases, hexokinase that binds to it is saturated, and excess glucose binds to aldose reductase, which is less affinity than hexokinase, and is metabolized to sorbitol to accumulate sorbitol in cells. Aldose reductase is present in the retina, sclera, lens, kidney, erythrocytes, brain, muscle, liver and peripheral nerves, especially in the peripheral nerves can be a problem because glucose permeation is not affected by insulin. In other words, when hyperglycemia accumulates too much sorbitol in Schumann cells, the osmotic pressure may increase, resulting in edema, necrosis and collapse of myelin, and water is introduced into the diabetic complications due to the continuously increased osmotic pressure. Et al., The Korean Diabetes Society, Korea Medicine, p483; Soulis-Liparota, T. et al., Diabetologia, 38, 357-394, 1995).

Final glycation end products have been reported to activate aldose reductase, a major enzyme in the polyol pathway, in human microvascular endothelial cells (Nakamura, N. et al., Free Radic Biol. Med., 29, 17-25 , 2000). In a steady state, aldose reductase has a very low affinity for glucose, but at high concentrations of glucose, aldose reductase is activated to excessively reduce glucose to sorbitol, and this sorbitol is converted to fructose by sorbitol dehydrogenase. This fructose is about 10 times faster than non-enzymatic glycosylation of protein. High concentrations of fructose bind to the protein, eventually accelerating the formation of the final glycated product. As diabetes increases the body's blood sugar for a long time, chronic complications occur as glycation products invade the kidneys, nerves, retinas, and large and small blood vessels throughout the body.

 Therefore, the extract extracted with alcohol from the ginseng of the present invention or 5-methylsophoraflavanone B separated therefrom exhibits inhibitory activity of aldose reductase, and thus can be used as an inhibitor of aldose reductase activity. It can also be used as a preventive and therapeutic agent for diabetes complications.

The diabetic complications are diabetic retinopathy, diabetic cataract, diabetic nephropathy, diabetic neuropathy and the like.

In another aspect, the present invention provides a pharmaceutical composition for the prevention and treatment of obesity containing a ginseng extract or 5-methylsophoraflavanone B isolated therefrom as an active ingredient.

In an experiment conducted to verify the obesity inhibitory effect of 5-methylsophoraflavone B isolated from the ethanol extract of Ginseng according to the present invention, 5-methyl vesicles in the process of inducing differentiation of undifferentiated 3T3-L1 cell line into adipocytes By treating with rabanon B, it was confirmed that there is an inhibitory effect on adipocyte differentiation (see FIGS. 1 , 2 and 3 ).

Therefore, the ginseng extract according to the present invention or 5-methylsophoraflavanone B isolated therefrom can be usefully used as an agent for preventing and treating obesity.

The 5-methylsophoraflavanone B of the present invention can be extracted from red ginseng or synthesized by an organic chemical synthesis method.

The composition may be used as a pharmaceutical composition, and may be various oral or parenteral formulations. When formulated, diluents or excipients such as fillers, extenders, binders, wetting agents, disintegrating agents, and surfactants are usually used. Solid form preparations for oral administration include tablets, pills, powders, granules, capsules, and the like, which form at least one excipient such as starch, calcium carbonate, sucrose or lactose (at least one compound). lactose) and gelatin. In addition to simple excipients, lubricants such as magnesium stearate, talc and the like are also used. Liquid preparations for oral administration include suspensions, liquid solutions, emulsions, and syrups, and various excipients such as wetting agents, sweeteners, fragrances, and preservatives, in addition to commonly used simple diluents such as water and liquid paraffin, may be included. have. Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized preparations, suppositories. As the non-aqueous solvent and the suspension solvent, propylene glycol, polyethylene glycol, vegetable oils such as olive oil, injectable esters such as ethyl oleate, and the like can be used. As the base of the suppository, witepsol, macrogol, tween 61, cacao butter, laurin butter, glycerogelatin and the like can be used.

The composition of the present invention may be administered parenterally or orally as desired, and may be administered in one to several times so as to be administered in an amount of 0.1 to 500 mg, preferably 1 to 100 mg per kg of body weight per day. have. The dosage for a particular patient may vary depending on the patient's weight, age, sex, health condition, diet, time of administration, method of administration, rate of excretion, severity of the disease, and the like.

In addition, the present invention provides a health food for preventing and improving diabetic complications containing a ginseng extract or 5-methylsophoraflavanone B isolated therefrom as an active ingredient.

The composition according to the present invention can be added to the health supplements, such as food, beverages and the like, Ginseng extract or 5-methylsophoraflavanone B isolated therefrom for the purpose of preventing and improving the complications of diabetes. In this case, the ginseng extract or 5-methylsophoraflavanone B isolated therefrom may be added in an amount of 0.01 to 20% by weight, preferably 0.1 to 5% by weight, based on the raw material when used as a food additive. The blending amount of the active ingredient may be appropriately determined depending on the purpose of use (prevention, health or therapeutic treatment). However, in the case of long-term intake for health and hygiene purposes or health control purposes, the amount may be below the above range, and since there is no problem in terms of stability, the active ingredient may be used in an amount above the above range. The extract or a compound separated therefrom may be used with other food or food ingredients, and may be appropriately used according to conventional methods.

There is no particular limitation on the kind of food. Examples of the food to which the extract or 5-methylsophoraflavanone B separated from the extract may be added include meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, gum, and ice cream. Dairy products, various soups, beverages, tea, drinks, alcoholic beverages and vitamin complexes, and the like, including all of the health food in the usual sense.

The food supplement additive of the present invention may use various flavors or natural carbohydrates. The natural carbohydrates are sugars such as glucose, monosaccharides such as fructose, disaccharides such as maltose and sucrose, polysaccharides such as dextrin and cyclodextrin, xylitol, sorbitol, and erythritol. As the sweetening agent, natural sweetening agents such as tautin and stevia extract, synthetic sweetening agents such as saccharin and aspartame, and the like can be used. The proportion of the natural carbohydrate is generally about 0.01 to 0.04 parts by weight, preferably 0.02 to 0.03 parts by weight, per 100 parts by weight of the composition according to the present invention.

In addition to the above, the composition according to the present invention includes various nutrients, vitamins, electrolytes, flavors, coloring agents, pectic acid and salts thereof, alginic acid and salts thereof, organic acids, protective colloidal thickeners, pH adjusting agents, stabilizers, preservatives, glycerin, Alcohols, carbonating agents used in carbonated drinks, and the like. In addition, the composition according to the present invention may contain a pulp for the production of natural fruit juices and vegetable drinks. These components can be used independently or in combination, and the proportion of the additive is not critical, but is usually selected in the range of 0.01 to 0.1 parts by weight per 100 parts by weight of the composition according to the present invention.

Hereinafter, the present invention will be described in detail through Examples and Experimental Examples. However, the following examples are merely to illustrate the present invention, but the content of the present invention is not limited by the following examples.

< Example  1> Preparation of Ginseng Extract

100 ml of ethanol was added to the ginseng root (20 g), and extracted by heating at a temperature of 70 to 80 ° C. for 1 hour, and only the ethanol soluble portion was recovered using a filter paper. After repeating this three times, the ethanol soluble part was concentrated under reduced pressure to obtain ethanol extract (2 g) of red ginseng.

< Example  2> Gosam  5- from extract Methyl Sophoraflavanone  Separation of B

The ethanol extract (2 g) obtained in Example 1 was dissolved in methanol, and then subjected to high performance liquid chromatography to perform not only 5-methylsophoraflavanone B but also 3,7,4'-trihydroxy-5-meth. Toxy-8-prenylflavanone, cusenol N, curarinone, 2'-methoxycurarinone, sophoraflavanone G, lycanone A and curaridine were isolated. The high performance liquid chromatography conditions are as follows. As a mobile phase, a mixed solvent of water and acetonitrile was used, and the conditions were used in a concentration gradient of 35% to 70% by volume acetonitrile for 60 minutes. The flow rate was 10 ml per minute and the column used HiQ sil C18 (φ 21.2 × 250 mm, 10 μm). The isolated compound was identified by spectroscopic method ( ¹ H-NMR spectroscopy (Varian unity Inova with cryoprobe, Varian, USA). The analysis results are shown in Table 1 .

compound Compound name ¹ H-NMR (DMSO-d ₆ ) One 5-methylsophoraflavanone B 7.28 (2H, d, J = 7.4 Hz, H-2 ', 6'), 6.77 (2H, d, J = 7.4 Hz, H-3 ', 5'), 6.14 (1H, s, H-6) , 5.32 (1H, doublet of doublets, J = 12.5, 3.0 Hz, H-2), 5.09 (1H, tt, J = 1.3, 7.0 Hz, H-2 "), 3.69 (3H, s, 5-OCH ₃ ), 3.11 (2H, d, J = 7.0 Hz, H-1 "), 2.92 (1H, dd, J = 16.4, 12.5 Hz, J = 3), 2.56 (1H, dd, J = 16.4, 3.0 Hz, H- 3), 1.58 (3H, s, H-5 "), 1.54 (3H, s, H-4") 2 3,7,4'-trihydroxy-5-methoxy-8-prenylflavanone 7.27 (2H, d, J = 8.3 Hz, H-2 ', 6'), 6.76 (2H, d, J = 8.3 Hz, H-3 ', 5'), 6.15 (1H, s, H-6) , 5.15 (1H, d, J = 4.3 Hz, 3-OH), 5.06 (1H, brt, J = 7.2 Hz, H-2 ″), 4.89 (1H, d, J = 11.3 Hz, H-2), 4.24 (1H, doublet of doublets, J = 11.3, 4.3 Hz, H-3), 3.71 (3H, s, 5-OCH ₃ ), 3.04 (2H, m, H-1 "), 1.55 (3H, s, H- 4 "), 1.46 (3H, s, H-5") 3 Cushenol N 7.12 (1H, d, J = 8.4 Hz, H-6 '), 6.31 (1H, d, J = 1.9 Hz), 6.22 (1H, dd, J = 8.4, 1.9 Hz, H-5'), 6.11 ( 1H, s, H-6), 5.21 (1H, d, J = 11.1 Hz, 3-OH), 5.04 (1H, d, J = 3.3 Hz, H-2), 4.86 (1H, brt, J = 6.5 Hz, H-4 "), 4.50 (1H, s, H-9" a), 4.43 (1H, s, H-9 "b), 4.36 (1H, dd, J = 11.1, 3.3 Hz, H-3 ), 3.71 (3H, s, 5-OCH ₃ ), 2.42-2.37 (3H, m, H-1 ", 2"), 1.90 (2H, m, H-3 "), 1.51 (6H, s, H -7 ", 10"), 1.44 (3H, s, H-6 ") 4 Kurarinon 7.22 (1H, d, J = 8.4 Hz, H-6 '), 6.34 (1H, d, J = 1.7 Hz, H-3'), 6.26 (1H, dd, J = 8.4, 1.7 Hz, H-5 '), 6.12 (1H, s, H-6), 5.43 (1H, dd, J = 13.0, 2.1 Hz, H-2), 4.90 (1H, brt, J = 5.8 Hz, H-4 "), 4.56 (1H, brs, H-9 "a), 4.49 (1H, brs, H-9" b), 3.71 (3H, s, 5-OCH ₃ ), 2.82 (1H, dd, J = 16.2, 13.0 Hz, H-3a), 2.51 (1H, dd, J = 13.0, 2.1 Hz, H-3b), 2.51-2.42 (3H, m, H-1 ", 2"), 1.93 (2H, m, H-3 " ), 1.58 (3H, s, H-10 "), 1.54 (3H, s, H-7"), 1.46 (3H, s, H-6 ") 5 2'-methoxycurinone 7.29 (1H, d, J = 8.2 Hz, H-6 '), 6.43 (1H, d, J = 1.2 Hz, H-3'), 6.11 (1H, s, H-6), 5.42 (1H, dd , J = 13.6, 1.8 Hz, H-2), 4.86 (1H, brt, J = 5.4 Hz, H-4 "), 4.54 (1H, brs, H-9" a), 4.46 (1H, brs, H -9 "b), 3.71 (3H, s, 2'-OCH ₃ ), 3.69 (3H, s, 5-OCH ₃ ), 2.84 (1H, dd, J = 16.1, 13.6 Hz, H-3a), 2.45 -2.39 (4H, m, H-3b, 1 ", 2"), 1.91 (2H, m, H-3 "), 1.56 (3H, s, H-10"), 1.51 (3H, s, H- 7 "), 1.41 (3H, s, H-6") 6 Sophoraflavanone G 7.27 (1H, d, J = 8.3 Hz, H-6 '), 6.39 (1H, d, J = 2.0 Hz, H-3'), 6.31 (1H, dd, J = 8.3, 2.0 Hz, H-5 '), 5.99 (1H, s, H-8), 5.55 (1H, dd, J = 13.1, 2.3 Hz, H-2), 4.95 (1H, brt, J = 6.0 Hz, H-4 "), 4.61 (1H, s, H-9 "a), 4.54 (1H, s, H-9" b), 3.16 (1H, dd, J = 17.2, 13.1 Hz, H-3a), 2.66 (1H, dd, J = 17.2, 2.3 Hz, H-3b), 2.54-2.44 (3H, m, H-1 ", 2"), 1.99 (2H, m, H-3 "), 1.62 (3H, s, H-10" ), 1.58 (3H, s, H-7 "), 1.49 (3H, s, H-6") 7 Lichnonon A 7.31 (1H, dd, J = 8.4, 1.9 Hz, H-6 '), 6.48 (1H, d, J = 1.9 Hz, H-3'), 6.41 (1H, dd, J = 8.4, 1.9 Hz, H -5 '), 5.93 (1H, s, H-6), 5.54 (1H, dd, J = 13.3, 3.0 Hz, H-2), 4.88 (1H, brt, J = 6.7 Hz, H-4 ") , 4.56 (1H, s, H-9 "a), 4.47 (1H, s, H-9" b), 3.74 (3H, s, 2'-OCH3), 3.18 (1H, dd, J = 17.1, 13.3 Hz, H-3a), 2.61 (1H, dd, J = 17.1, 3.0 Hz, H-3b), 2.47-2.32 (3H, m, H-1 ", 2"), 1.97 (2H, m, H- 3 "), 1.55 (3H, s, H-10"), 1.52 (3H, s, H-7 "), 1.43 (3H, s, H-6") 8 Curaridine 14.92 (1H, s, 9-OH), 7.97 (1H, d, J = 15.5 Hz, H-2), 7.88 (1H, d, J = 15.5 Hz, H-3), 7.46 (1H, d, J = 8.5 Hz, H-6 '), 6.40 (1H, d, J = 2.1 Hz, H-3'), 6.34 (1H, dd, J = 8.5, 2.1 Hz, H-5 '), 6.07 (1H, s, H-6), 5.00 (1H, brt, J = 5.5 Hz, H-4 "), 4.60 (1H, brs, H-9" a), 4.53 (1H, brs, H-9 "b), 3.86 (3H, s, 5-OCH ₃ ), 2.57-2.44 (3H, m, H-1 ", 2"), 2.03 (2H, m, H-3 "), 1.68 (3H, s, H-10 "), 1.62 (3H, s, H-7"), 1.53 (3H, s, H-6 ")

< Experimental Example  1> Gosam  Of extract Aldos  Inhibitory Effect of Reductase

In order to measure the inhibitory effect of the aldose reductase activity of the composition according to the present invention was carried out experiments on the enzyme precipitate of the lens of the rat. First, in order to obtain an aldose reductase source, the lens of the rat was extracted, and a certain amount of phosphate buffer was added according to the amount of wet growth, and the lens was homogenized. The homogenate was centrifuged at 4 ° C. to obtain a supernatant, and ammonium sulphate was added to the supernatant so that the final concentration was 40%. Thereafter, the supernatant was collected by centrifugation again, and then ammonium sulfate was added thereto at a final concentration of 70%, followed by mixing for about one hour. The precipitate obtained by centrifugation was suspended in PBS buffer and dialyzed for 1 day to obtain an aldose reductase source. Thereafter, 10 g / ml concentration of high ginseng extract was added to the prepared aldose reductase source, and the reaction rate of DL-glyceraldehyde was reacted with a substrate to measure NADPH absorbance reduction rate at 340 nm. As a control, tetramethylene glutaric acid was used.

As a result of the treatment, 10 g / ml of Ginseng extract showed 63% inhibition of aldose reductase activity compared to the untreated control, and it was confirmed that Ginseng extract has pharmacological activity that inhibits aldose reductase activity. .

< Experimental Example  2> Gosam  Of compounds isolated from extracts Aldos  Inhibitory Effect of Reductase

First, an aldose reductase source was obtained in the same manner as in Experiment 1. 5-methylsophoraflavanone B (Compound 1) obtained in Example 2, 3,7,4'-trihydroxy-5-methoxy-8-prenylflavanone (compound) 2), Cushenol N (Compound 3), Curarinone (Compound 4), 2'-Methoxycurinone (Compound 5), Sophoraflavanone G (Compound 6), Lirichanone A (Compound 7) and Curaridine (Compound 8) was added to each other, and DL-glyceraldehyde was reacted with a substrate to measure the rate of decrease of NADPH absorbance at 340 nm. As a control, tetramethylene glutaric acid was used. The measurement results are shown in Table 2 below.

compound Concentration (m) % Inhibition IC ₅₀ (M) One 100 94.90 0.29 10 80.98 One 75.92 0.1 30.36 2 100 93.63 0.54 10 73.39 One 50.61 0.1 39.22 3 100 79.71 1.55 10 62.12 One 55.68 0.1 24.04 4 100 96.16 1.61 10 64.53 One 35.43 0.1 30.36 5 100 87.31 1.48 10 60.74 One 43.02 0.1 32.90 6 100 100.0 0.56 10 89.83 One 51.88 0.1 31.63 7 100 79.72 3.79 10 60.74 One 31.36 0.1 21.51 8 100 99.96 0.99 10 75.92 One 45.82 0.1 27.84 Control 100 91.10 2.45 10 72.12 One 36.69

As shown in Table 2 , all of the compounds isolated from the extract of Ginseng showed better or comparable inhibitory activity than tetramethylene glutaric acid used as a control. In particular, 5-methylsophoraflavanone B of the formula (1) of the present invention showed the best inhibitory activity against aldose reductase, the IC ₅₀ value was 0.29.

< Experimental Example  3> Gosam  5- isolated from extract Methyl Sophoraflavanone  B obesity suppression

In order to verify the differentiation inhibitory effect of the undifferentiated 3T3-L1 cell line (ATCC) into adipocytes using 5-methylsophoraflavanone B isolated from the extract The same experiment was performed.

Undifferentiated 3T3-L1 cell lines were maintained in DMEM medium containing 10% bovine calf serum. The undifferentiated 3T3-L1 cell line was cultured to a density of about 90%, and then DMEM medium containing 10% fetal bovine serum, dexamethasone, IBMAX, and insulin was replaced to induce differentiation. In the case of medium replacement, 100 μM of 5-methylsophoraflavanone B dissolved in dimethyl sulfoxide (DMSO) was continuously treated. After incubation for 2 days, the cells were replaced with DMEM medium containing 10% fetal calf serum and insulin. It was then replaced with DMEM medium containing only 10% fetal calf serum every two days. Eight days after the induction of regeneration, the regeneration into adipocytes was complete. At this time, the control group was replaced with the medium at exactly the same interval as the experimental group, but treated only the same concentration of DMSO. After redifferentiation was completed, appearance photographs and micrographs were stained with oil red o, which selectively stained only lipid components in red. At the same time, oil red o dye was extracted from the cells, and then the absorbance (520 nm) of the solution was measured by spectrophotometer to compare the degree of differentiation into adipocytes. The experimental results are shown in FIGS. 1, 2 and 3.

As shown in FIG. 1 , the control group (a) was found to have a large number of fat cells stained with oil red, whereas the 3T3-L1 cell line was treated with 5-methylsophoraflavanone B during differentiation into adipocytes (b ), There are few differentiated fat cell lines. As shown in FIG. 2 , it was confirmed that 3T3-L1 cell line significantly inhibited fat cell formation by 50% when treated with 5-methylsophoraflavanone B during the differentiation into adipocytes. In addition, as shown in FIG . 3 , the results of microscopic observation showed that adipocyte differentiation was significantly suppressed when 5-methylsophoraflavanone B treatment (b) was compared with the control group (a).

< Formulation example  1> Preparation of Pharmaceutical Formulations

1-1. Powder  Produce

Ginseng extract or 5-methylsophoraflavanone B 2 g isolated therefrom

1 g lactose

After mixing the above components, the airtight cloth was filled to prepare a powder.

1-2. Manufacture of tablets

Gosam extract or 5-methylsophoraflavanone B isolated from 100 mg

100 mg corn starch

Lactose 100 mg

2 mg magnesium stearate

After mixing the above components, tablets were prepared by tableting according to a conventional method for producing tablets.

1-3. Preparation of Capsules

Gosam extract or 5-methylsophoraflavanone B isolated from 100 mg

100 mg corn starch

Lactose 100 mg

2 mg magnesium stearate

After mixing the above components, the capsule was prepared by filling in gelatin capsules according to the conventional method for producing a capsule.

1-4. Preparation of Injection

10 ㎍ / mL Gosam extract or 5-methylsophoraflavanone B isolated therefrom

Dilute hydrochloric acid BP until pH 3.5

Injectable sodium chloride BP up to 1 ml

Sodium Ginseng extract or 5-methylsophoraflavanone B isolated therefrom was dissolved in an appropriate volume of sodium chloride BP for injection, and the pH of the resulting solution was adjusted to pH 3.5 using dilute hydrochloric acid BP, and the sodium chloride BP for injection was Volume to adjust and thoroughly mix. The solution was filled into a 5 ml Type I ampoule made of clear glass, encapsulated under an upper grid of air by dissolving the glass, and sterilized by autoclaving at 120 ° C. for at least 15 minutes to prepare an injection solution.

< Production Example  2> Manufacture of health food

2-1. Manufacture of beverages

522 mg of honey

Chioctosanamide 5 mg

Nicotinamide 10 mg

Riboflavin Sodium Hydrochloride 3 mg

Pyridoxine hydrochloride 2 mg

Inositol 30 mg

Orthoic acid 50 mg

Red ginseng extract or 5-methylsophoraflavanone B 0.48-1.28 mg isolated therefrom

200 ml of water

With the above composition and content, drinks were prepared using conventional methods.

2-2. Chewing gum  Produce

Gum base 20%

Sugar 76.36-76.76%

Red ginseng extract or 5-methylsophoraflavanone B isolated from 0.24 to 0.64%

1% fruit flavor

Water 2%

Chewing gum was prepared using conventional methods using the above composition and content.

2-3. Candy

50-60% sugar

Starch syrup 39.26 ~ 49.66%

Red ginseng extract or 5-methylsophoraflavanone B isolated from 0.24 to 0.64%

Orange flavor 0.1%

Candy was prepared using conventional methods with the above composition and content.

As described above, according to the present invention, 5-methylsophoraflavanone B, a single compound isolated from the extract of Ginseng, shows an inhibitory effect on the activity of aldose reductase, which causes diabetic complications, and also inhibits adipocyte differentiation. By preventing obesity, the composition according to the present invention can be usefully used for preventing or treating diabetes complications or obesity.

Claims (10)

Group consisting of 3,7,4′-trihydroxy-5-methoxy-8-prenylflavanone, cusenol N, curarinone, 2′-methoxycurarinone, sophoraflavanone G, and curaridine A pharmaceutical composition for preventing and treating diabetic retinopathy, diabetic cataract, diabetic nephropathy or diabetic neuropathy, containing any one or more compounds selected from the group as an active ingredient. A health food for preventing and improving diabetic retinopathy, diabetic cataract, diabetic nephropathy or diabetic neuropathy, comprising the compound of claim 1 as an active ingredient. delete delete delete delete delete delete delete delete

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