JP2006151878A - Hypoglycemic agent for diabetes and remedy for hyperlipemia - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a hypoglycemic agent for diabetes, having safety and high efficacy, and to provide a remedy for hyperlipemia. <P>SOLUTION: This hypoglycemic agent for the diabetes contains curcumin and further contains one or more kinds of xanthine derivatives selected from caffeine, theophylline, and theobromine. Further, the hypoglycemic agent for the diabetes contains a rhizomatous extract of a plant belonging to the Curcuma genus of the Zingiberaceae family and further contains one or more kinds of the xanthine derivatives selected from the caffeine, the theophylline, and the theobromine. Thus, action only by the curcumin, or only by the extract of the plant belonging to the Curcuma genus of the Zingiberaceae family, is enhanced in the hypoglycemic agent by together using the xanthine derivatives. <P>COPYRIGHT: (C)2006,JPO&NCIPI

Description

  The present invention relates to a diabetic hypoglycemic agent and an antihyperlipidemic agent containing curcumin and a xanthine derivative as active ingredients.

  The rhizomes of the ginger family Curcuma genus (Curcuma genus) are used as edible as raw materials for curry powder, and have been used for a long time as a traditional Chinese medicine such as a healthy stomach and galling agent. One of the main pharmacological components contained in this turmeric is curcumin (1,7-bis- (4-hydroxy-3-methoxyphenyl) hepta-1,6-diene-3,5-dione), which is antidepressant Agents, anti-menopausal agents (see Patent Document 1), strong muscle agents, anti-inflammatory agents (see Patent Document 2), blood flow improving compositions (see Patent Document 3), hematopoietic tumor therapeutic agents, immunosuppressants (See Patent Document 4), an anti-aging agent (Patent Document 5), and the like, and also applied to the prevention and treatment of lifestyle-related diseases that have recently been increasing. For example, drugs based on hypoglycemic action in type 2 diabetes model mice (see Patent Document 6) and inhibitory action on triglyceride accumulation in the liver of high-fat diet-fed mice (see Patent Document 7) are known. .

JP 2003-113117 A JP 2004-182599 A JP 2004-123707 A JP 2004-75666 A Japanese Patent Laid-Open No. 2004-2237 Japanese Patent Laid-Open No. 2003-128539 Japanese Patent Laid-Open No. 11-246399

  However, the action of curcumin is extremely weak compared to synthetic antidiabetic drugs such as thiazolidinedione and fibrate synthetic antilipidemic drugs. However, turmeric extract and curcumin are excellent in safety, as can be seen from the fact that they are also foodstuffs, and there is a need for a method that enhances the hypoglycemic effect while taking advantage of this feature.

  Under these circumstances, as a result of intensive studies, the present inventors have enhanced the blood glucose lowering effect and neutral fat reducing effect of turmeric and curcumin by the combined use of xanthine derivatives usually contained in tea and coffee such as theophylline and caffeine. As a result, the present invention has been completed. That is, the present invention includes curcumin and curcumin-containing turmeric extract, and further contains one or more xanthine derivatives selected from caffeine, theophylline, or theobromine, a hypoglycemic agent for diabetes, Or it relates to a therapeutic agent for hyperlipidemia.

  In the present invention, diabetic blood glucose, blood triglyceride or blood total cholesterol reduction by curcumin or turmeric extract is synergistically enhanced by a simple method using a xanthine derivative such as caffeine. This synergistic effect is that the inhibition of phosphodiesterase by xanthine derivatives increases the cAMP level of second messenger in the body, which is coupled with the activation of PPAR transcription factors by curcumin to increase production and activation of regulators of lipid and energy metabolism It is thought to do.

  As such regulatory factors, adiponectin, a cytokine whose deficiency is closely related to the development of lifestyle-related diseases (Non-patent literature: Ichiro Shimomura et al., Experimental Medicine, Vol. 20, No. 12, 1762-1767 ( 2002)). As shown in the Examples described later, simultaneous administration of curcumin and caffeine increases the effective concentration of adiponectin in blood in inverse correlation with the decrease in blood glucose level. This suggests that the diabetic hypoglycemic and hyperlipidemic effects of the composition containing both curcumin and caffeine according to the present invention are based on the enhancement of adiponectin. In the present invention, the diabetic hypoglycemic and hyperlipidemic effects of the composition containing both curcumin and caffeine are also exerted in actual animals, and these actions are related to the adiponectin enhancing action. This is the first time to clarify this at the level of actual animal experiments. Furthermore, since caffeine, which is one of the active ingredients of the present invention, is a compound that has been conventionally eaten and consumed, it can be said that the drug of the present invention based on these combinations is extremely safe.

  Curcumin used in the present invention is extracted and purified from turmeric rhizomes by a known method, and can be easily chemically synthesized by a known method. On the other hand, since turmeric contains many pharmacologically active compounds in addition to curcumin, it is also preferably used in the present invention as a crude extract (extract). For the preparation of such an extract, the rhizomes of turmeric (Curcuma longa) and spring turmeric (Curcuma aromatica) are kept fresh or dried, chopped and ground, and then extracted with a suitable solvent such as ethanol. The residue obtained by concentrating the extract and removing the solvent can be used as it is, but it may be used after removing unnecessary components and activity-inhibiting components by known means such as activated carbon treatment or silica gel column chromatography. On the other hand, the xanthine derivative used in combination with curcumin or turmeric extract is one or a mixture of two or more of theophylline, caffeine, or theobromine. These can be extracted from tea leaves, coffee beans, cacao beans, etc., and can be easily synthesized by known methods.

  In addition, these xanthine derivatives can be used as raw materials such as tea leaves or in the state of a crude extract as long as there is no inconvenience in the activity. As a method of using these xanthine derivatives, curcumin, or a crude extract thereof, a method of dissolving in ethanol or water and adding to drinking water, a method of suspending in water containing a surfactant such as an appropriate phospholipid, In addition to adding to food, there may be mentioned a method of mixing with an appropriate adjuvant and using it in the form of a tablet. The amount of curcumin added for use in the present invention is 1 mg to 10 mg or less per 1 ml of drinking water or 1 g of food, but it may be appropriately adjusted depending on the case. The amount of turmeric extract added is appropriately adjusted in order to obtain the desired effect within a physiologically safe range depending on the method and degree of purification. The amount of xanthine derivative added is in accordance with the concentration contained in commonly used coffee and green tea. Caffeine is 0.1 to 0.5 mg (about 3 mM) per gram of drinking water or 1 g of food, theophylline is 0.01 to 0.05 mg (about 0.3 mM) per liter of drinking water or 1 g of food, and theobromine is drinking water. The standard is 0.1 mg to 2 mg (about 10 mM) per gram of food or 1 g of food, but the dosage should be adjusted appropriately within a physiologically safe range depending on the total intake and intake form. Next, the present invention will be described in more detail based on examples.

[Example 1]
KK-Ay / Ta Jc1 mice genetically developing type 2 diabetes (male, 5 weeks old, 17 mice, manufactured by Claire Japan) with a feed prepared with or without curcumin and / or caffeine Breeding for 4 weeks, body weight and food consumption were measured regularly, and blood was collected once a week to measure blood glucose level, blood adiponectin level, plasma triglyceride and total cholesterol concentration. The feed is 4 ml of ethanol suspension of Sigma's curcumin (0.15 g / ml) and / or 2 ml of aqueous caffeine (30 mg / ml) per 120 g of CE-2 powder feed made by CLEA Japan. In addition, 2.4 g of potato starch was mixed with 48 ml of water and heated, and the whole was mixed well with a mortar and pestle, and then filled firmly in a petri dish with a diameter of 15 cm. The solid feed prepared by air drying was freely ingested. Water was ingested freely by sterilizing tap water with an autoclave.

  Plasma (plasma) for blood glucose and adiponectin measurement is obtained by collecting 9 μl of blood from the tip of the mouse tail, and immediately mixing this with 3 μl of heparin solution (50 mg / ml) and centrifuging (12000 rpm, 12 minutes) ) To remove blood cells. The concentrations of glucose, triglyceride and total cholesterol in the plasma were measured with glucose CII-Test Wako, Triglyceride E-Test Wako and Cholesterol E-Test Wako (all manufactured by Wako Pure Chemical Industries, Ltd.). On the other hand, 2 μl of plasma was mixed with 8 μl of water and 10 μl of 2 × SDS buffer, and then heat-treated at 95 ° C. for 5 minutes to prepare an electrophoresis sample. Detection of adiponectin by electrophoresis using polyacrylamide gel (applying 2 μl of each lane) and Western blotting was performed by a conventional method.

  The primary antibody used was mouse anti-adiponectin, mouse monoclonal antibody (Catalog No. MAB3608) manufactured by Chemicon, and the secondary antibody was rabbit anti-mouse Ig G1-HRP conjugate (Catalog No. 61- 0120). For detection of the band, an ECL Plus measurement kit manufactured by Pharmacia was used. Moreover, about the gel after electrophoresis, the part with a molecular weight of about 80,000 was dye | stained with the CBB reagent, and serum albumin was detected. These bands were loaded into a personal computer as an image with a scanner, and the density of the band was digitized with Gel Pro Analyzer (software). From these values, the relative level of adiponectin in blood based on serum albumin was determined.

  The results are shown in Table 1, Table 2, and FIGS. As shown in Table 1, there is no significant difference in the average body weight of each group of mice. Table 2 shows that the feed (curcumin + caffeine group) in which both curcumin and caffeine are mixed is somewhat less ingested than the other groups.

注１）；クルクミン及び/又はカフェインを添加した飼料で飼育したKK-Ay/Ta Jc1マウスの飼育期間中における体重増加を示す。マウスの数はコントロール群で５匹、他の群は各４匹、数値は平均値と標準偏差である。

Note 1): Shows the increase in body weight during the breeding period of KK-Ay / Ta Jc1 mice raised on a diet supplemented with curcumin and / or caffeine. The number of mice is 5 in the control group, 4 in each of the other groups, and the values are the mean and standard deviation.

注２）；クルクミン及び/又はカフェインを添加した飼料で飼育したKK-Ay/Ta Jc1マウスの飼育期間中における1日１匹あたりの平均摂餌量を示す。各群の1週間における総摂餌量から計算した。数値は平均値と標準偏差である。

Note 2): Shows the average amount of food consumed per day during the breeding period of KK-Ay / Ta Jc1 mice raised on a diet supplemented with curcumin and / or caffeine. Calculations were made from the total food intake during one week for each group. Numbers are mean and standard deviation.

  FIG. 1 shows that plasma glucose levels are lowest in the curcumin + caffeine group for 8-10 weeks of age, and that the increase in blood glucose due to the onset of diabetes is most effectively suppressed by simultaneous administration of curcumin and caffeine. [The curcumin + caffeine group shows a significantly lower value than the control group at each age (p <0.05)]. Fig. 2 shows the measurement results of blood adiponectin at 9 and 10 weeks of age. Compared to the control group, the blood adiponectin level increased with the addition of curcumin feed, and curcumin plus caffeine in addition to curcumin + It was shown that the level was further increased in the caffeine group (p <0.05, curcumin group vs. curcumin + caffeine group). In this case, even when only caffeine was added, the level of adiponectin was higher than that of the control, but in FIG. 1, it was considered that the blood glucose level was the same as that of the control at 9, 10 weeks of age with caffeine alone. Adiponectin, which is effective for lowering blood sugar, is interpreted to be increased by co-administration with curcumin. Furthermore, KK-Ay / Ta mice are also models of hyperlipidemia because blood fat increases with the progression of diabetes and obesity (Japanese Patent Laid-Open No. 2004-168720). In the results of the measurement of plasma triglycerides and total cholesterol in this experiment (Fig. 3), the curcumin + caffeine group showed the lowest value. Compared with curcumin alone, simultaneous administration of curcumin and caffeine resulted in blood fat and cholesterol. The concentration was further reduced (p <0.05).

[Example 2]
200 g of dried turmeric rhizome powder (manufactured by Konishi Pharmaceutical) is mixed with 1 liter of hexane, stirred for 2 days at room temperature with a magnetic stirrer, turmeric powder is collected by filtration, and a 1: 1 mixture of acetone and hexane The mixture was further stirred and extracted for 1 day with 1 liter. The extract obtained by filtering the powder was concentrated with a rotary evaporator and dried under reduced pressure with an oil pump to obtain 11.1 g of an oily substance.

注３）；ウコンエキスとカフェインを添加した飼料で飼育したKK-Ay/Ta Jc1マウスの飼育期間中における体重増加を示す。マウスの数はコントロール群とウコンエキス＋カフェイン群で各３匹、ウコンエキス群で４匹、数値は平均値と標準偏差である。

注４）；ウコンエキスとカフェインを添加した飼料で飼育したKK-Ay/Ta Jc1マウスの飼育期間中における1日１匹あたりの平均摂餌量を示す。各群の1週間における総摂餌量から計算した。数値は平均値と標準偏差である。

表３に示すように、ウコンエキスのみの群では体重増加がコントロール群より大きかったが、ウコンエキス＋カフェイン群はコントロール群との間に大きな差はなかった。一方、摂餌量(表4)については、ウコンエキス＋カフェイン群がコントロール群に比べてわずかながら減少傾向を示した。図４は血糖測定の結果であるが、８、９週令目の血糖値がコントロール群やウコンエキス群に比べウコンエキス＋カフェイン群で顕著に減少していることを示す。 Example 3
KK-Ay / Ta Jc1 mice (male, 5 weeks old, 10 mice) were bred for 4 weeks with the turmeric extract prepared in Example 2, the feed with added turmeric extract and caffeine, or the feed without these added. Body weight, food intake and blood glucose level were measured regularly. The feed is 1.2 ml of the above turmeric extract ethanol solution (0.2 g / ml) per 120 g of CE-2 powder feed, or 1.2 ml of the same ethanol solution and 2 ml of caffeine aqueous solution (30 mg / ml), or ethanol only 1.2 ml of was added, and the powdered feed mixed well in the mortar was ingested freely. Water was ingested freely by sterilizing tap water with an autoclave. The blood glucose level was measured by using a glucose sensor (Glutest Neo sensor manufactured by Sanwa Chemical Laboratory) after collecting a small amount of blood from the tail of the mouse. The results are shown in Table 3, Table 4, and FIG.

Note 3): Shows the increase in body weight during the breeding period of KK-Ay / Ta Jc1 mice raised on feed supplemented with turmeric extract and caffeine. The number of mice is 3 for each of the control group and the turmeric extract + caffeine group, 4 for the turmeric extract group, and the numerical values are the mean and standard deviation.

Note 4): Shows the average amount of food consumed per day during the breeding period of KK-Ay / Ta Jc1 mice fed with feed supplemented with turmeric extract and caffeine. Calculations were made from the total food intake during one week for each group. Numbers are mean and standard deviation.

As shown in Table 3, the weight gain was greater in the turmeric extract alone group than in the control group, but the turmeric extract + caffeine group was not significantly different from the control group. On the other hand, as for food intake (Table 4), the turmeric extract + caffeine group showed a slightly decreasing tendency compared to the control group. FIG. 4 shows the results of blood glucose measurement, and shows that blood glucose levels at 8 and 9 weeks of age are significantly decreased in the turmeric extract + caffeine group compared to the control group and the turmeric extract group.

Example 4
KK-Ay / Ta Jc1 mice (male, 5 weeks old, 13 mice) were added with the turmeric extract prepared in Example 2 or it and caffeine, and the feed was solidified in the same manner as in Example 1 [Additional amount of turmeric extract Is 0.3% (w / w) of CE-2 powder] or solid feed without these supplements for 4 weeks, and body weight, food intake and plasma triglyceride concentration were measured regularly (control group and turmeric extract group) 4 each, 5 in turmeric extract + caffeine group). The changes in body weight and food intake were similar to the results of Example 3. The results of measurement of plasma triglyceride concentration are shown in FIG. 5. The increase in blood fat tended to be more strongly suppressed in the turmeric extract + caffeine group than in the turmeric extract group (p <0.05, 8 weeks old). Eye control group vs. turmeric extract group; p <0.10, turmeric extract group vs. turmeric extract + caffeine group).

Example 5
Another type 2 diabetes model mouse (BKS.C _g- + Lepr ^db / + Lepr ^db / Jc1, manufactured by CLEA Japan, 6 weeks old, 12 mice) was added with curcumin and / or caffeine in the same manner as in Example 1. The animals were reared for 4 weeks on a solid feed prepared without or added, and body weight, food intake, and blood glucose level were measured regularly (3 animals in 4 groups in total). There was no significant difference in body weight and food intake between groups. The measurement results of plasma glucose concentration are shown in FIG. 6. The group administered simultaneously with curcumin and caffeine showed a lower value than the control group throughout each week, and was lower than the group administered with curcumin alone at 10 weeks of age ( p <0.05).

It is a graph which shows the result of having measured the blood glucose level in 8-10 weeks old of the KK-Ay / Ta Jc1 mouse reared with the feed which added curcumin and / or caffeine. In the figure, the bar graph is the average value, the error bar is the standard deviation, and n = 9-12. It is a graph which shows the result of having measured the blood adiponectin level in 9 and 10-week-old of the KK-Ay / Ta Jc1 mouse reared with the feed which added curcumin and / or caffeine. In the figure, the bar graph is the average value, the error bar is the standard deviation, and n = 13 to 29. It is a graph which shows the result of having measured the plasma triglyceride density | concentration (A) and the total cholesterol density | concentration (B) in 9 and 10-week-old of the KK-Ay / Ta Jc1 mouse | mouth raised with the feed which added curcumin and / or caffeine. In the figure, the bar graph is the average value, the error bar is the standard deviation, and n = 9-12. It is a graph which shows the result of having measured the blood glucose level in 6-9 weeks old of the KK-Ay / Ta Jc1 mouse | mouth raised with the feed which added the turmeric extract and caffeine. In the figure, the bar graph is the average value, the error bar is the standard deviation, and n = 4,3. It is a graph which shows the plasma triglyceride density | concentration in 7-9 week-old of the KK-Ay / Ta Jc1 mouse | mouth raised with the feed which added the turmeric extract and caffeine. In the figure, the bar graph is the average value, the error bar is the standard deviation, and n = 8-15. It is a graph showing the blood glucose levels in ^{_{BKS.C g- + Lepr db / + Lepr}} 7~10 weeks old ^db / Jc1 mice fed with diets supplemented with turmeric extract. In the figure, the bar graph is the average value, the error bar is the standard deviation, and n = 6-9.

Claims (4)

  A diabetic hypoglycemic agent comprising curcumin and further containing one or more xanthine derivatives selected from caffeine, theophylline, or theobromine.   A diabetic hypoglycemic agent comprising a rhizome extract of a ginger family Curcuma genus, and further containing one or more xanthine derivatives selected from caffeine, theophylline, or theobromine.   A therapeutic agent for hyperlipidemia, comprising curcumin, and further containing at least one xanthine derivative selected from caffeine, theophylline, or theobromine. A therapeutic agent for hyperlipidemia, comprising a rhizome extract of a turmeric genus Curcuma, further containing at least one xanthine derivative selected from caffeine, theophylline, or theobromine

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