JP2000078956A - Food and medicine - Google Patents

Abstract

PROBLEM TO BE SOLVED: To obtain food and medicine for preventing or curing hypertension, allergy, cancer, hypercholesterolemia, hyperlipemia, etc., by containing the transglucosylated compound of a biologically active flavonoid. SOLUTION: In food and medicine, the transglucosylated compound of a physiologically active flavonoid is contained. As the biologically active compound, hesperidin, diosmin, naringin, neohesperidin, daidzin and genistin are cited. The transglucosylated compound of the physiologically active flavonoid is improved in absorptivity in living bodies, capable of expressing pharmacodynamic effects of the flavonoid and administered in a small dose owing to its improved absorptivity.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention is used as a food or drink or a medicine for preventing or treating hypertension, allergy, cancer, hypercholesterolemia, hyperlipidemia and the like.

[0002]

2. Description of the Related Art Numerous flavonoid compounds having various physiological activities are known in nature. However, it has been known that baicalein, which is one of the flavonoid compounds, has a very low absorption efficiency in the body, such that only about 1 / 3,000,000 of the dose is absorbed (Y. Wakui, et al.).
al. , J. et al. Chromatog. , 575,131
-136 (1992)). Therefore, the physiological activity of the flavonoid compound is not sufficiently exhibited in vivo. Therefore, it has been desired to increase the absorption efficiency.

As a result of intensive studies, the applicant of the present invention has found that the use of a bioactive flavonoid as a glycosyltransferred compound in which a sugar is bound to a bioactive flavonoid greatly improves the absorption efficiency in living organisms. In other words, it has been clarified for the first time that even a low-absorbing physiologically active flavonoid compound can be absorbed into the body at a high rate and quickly by using a glycosyltransferred compound and exert its effect for the first time.
Therefore, a remarkable effect can be expected with a small amount as compared with the conventional one.

[0004] These flavonoids are known to have, for example, the following physiological activities. Hesperidin and rutin have long been known as vitamin P to lower blood pressure (Shintaro Kamiya, New Vitamin Studies, (Japan Vitamin Society) 1969, p439). Hesperidin has an antiallergic effect (Hideaki Matsuda et al .; Pharmaceutical Magazine, 11
1, 193-198 (1991); A. Da Si
lva Emim et al. J .; Pharm. Ph
armacol. , 46, 118-712 (199).
4)), action to reduce LDL-cholesterol and improve blood cholesterol level (MT Monforte)
et al. Il Farmaco, 50, 595;
-599 (1995)), an anticancer effect (T. Tanak).
a, et al. Cancer Research,
54, 4653-4659 (1994); Tana
ka, et al. ; Cancer Research
h, 57, 246-252 (1997) T.H. Tanak
a, et al. Carcinogenesis, 1;
8, 761-769 (1997); Tanaka,
et al. Carcinogenesis, 18, 9;
57-965 (1997)).

[0005] In recent studies, hesperidin is also expected to promote the differentiation of preadipocytes and to improve symptoms such as diabetes. Diosmin has a strong antioxidant activity and a drug coexisting with hesperidin has been used as a therapeutic agent for venous insufficiency and hemorrhoids (C. La).
brid; Ancology, 45, 524-530.
(1994)). Furthermore, it has been reported that hesperidin, diosmin alone and a mixture thereof suppress oral cancer, esophageal cancer, colorectal cancer and the like (T. Tanak).
a, et al. Cancer Research,
54, 4653-4659 (1994); Tana
ka, et al. ; Cancer Research
h, 57, 246-252 (1997) T.H. Tanak
a, et al. Carcinogenesis, 1;
8, 761-769 (1997); Tanaka,
et al. Carcinogenesis, 18,
957-965 (1997)). Naringin and neohesperidin are known as citrus bitter substances, and are used in foods and beverages for the purpose of imparting bitterness.

[0006] More recently, it has been revealed that isoflavones are effective in improving bone density and that breast cancer is suppressed (Toda et al .; Food and Development). And the food and drink and the medicine which applied it were developed, and this invention was able to be completed. Hereinafter, the present invention will be described in detail.

[0007]

In the present invention, the in vivo absorption of a bioactive flavonoid compound and its glycosyltransferred compound was measured as follows.

A 5-week-old male St reared for 1 week
d. A ddY mouse (body weight, about 30 g) was fasted all day and night, and a predetermined concentration of a bioactive flavonoid compound or
300 μl of the glycosyltransferred compound solution was directly administered to the stomach using a probe. Controls received distilled water. After administration,
Blood was collected over time. The obtained blood was obtained from J.I. A. Bouti
n et al.'s method (The American society)
y for Pharmacology and Ex
peripheral Therapeutics, 2
1,1157-1166 (1993)), and then the physiologically active flavonoid compound or its glycosyltransfer compound contained therein was quantified using HPLC.

J. A. The method of Boutin et al. Is as follows. Acetonitrile (1000 μl) was added to the collected blood (500 μl), shaken sufficiently, and allowed to stand for 15 minutes. After centrifugation at 5500 rpm for 20 minutes, the supernatant was taken and freeze-dried. The lyophilized sample was subjected to acetonitrile / distilled water (20/80; v / v, 0.01M
NaOH) solution and analyzed by HPLC. HPLC conditions are as follows. column:
ODS, column temperature: 40 ° C., eluent: acetonitrile / distilled water (20/80; v / v), flow rate: 0.5 ml /
min, detection: UV 280 nm. The HPLC conditions for analyzing the isoflavone compound are as follows.
Column: ODS, column temperature: 40 ° C., eluent: gradient from acetonitrile / distilled water (15/85; v / v) to acetonitrile / distilled water (40/60; v / v), flow rate: 0.5 ml / min, detection: UV 280n
m. The concentration of the physiologically active flavonoid compound or its derivative in the blood was determined by preparing a calibration curve using a physiologically active flavonoid compound of known concentration or its glycosyltransferred compound.

[0010] In addition to mice, rats and the like can be used as experimental animals. In addition, the conditions of HPLC are slightly different depending on the physiologically active flavonoid compound and its transglycosylation compound to be measured, but the basic conditions are shown (individually described in Examples).

In the present invention, hesperidin, diosmin, naringin and neohesperidin are used as bioactive flavonoids, but isoflavones and the like can also be used. That is, flavonols, flavanones,
So-called flavonoids such as anthocyanidins and chalcones including isoflavones can be used.

[0012] Enzymatic glycosyltransfer reactions and chemical reactions can be used to produce glycosyltransferred compounds of bioactive flavonoids.
Any glycosyltransferase can be used for the glycosyltransferase. For example, it can be carried out using cyclodextrin synthase, amylase, etc., and a flavonoid glycosyltransfer compound can be obtained (T. Kometa).
ni, et al, Biosci. Biotech. B
iochem. , 58, 517-520 (1994).
T. Kometani, et al, Biosci. B
iotech. Biochem. , 58, 1990-1.
994 (1994). T. Kometani, et a
1, Biosci. Biotech. Bioche
m. , 60, 645-649 (1996). ). Also,
At this time, the origin of the enzyme is not limited.

[0013]

EXAMPLES Example 1 Five-week-old male Std. A ddY mouse (body weight, about 30 g) was fasted overnight, and 300 μl of a 20% (w / v) hesperidin transglycosylation compound and hesperidin solution was directly administered to the stomach using a probe. Controls received distilled water. After administration, blood was collected over time. The obtained blood was obtained from J.I. A. Bou
After pretreatment according to the method of Tin et al., the transglycosylation compound of hesperidin and hesperidin contained therein were quantified using HPLC. As a result, as shown in FIG. 1, the blood glycosyltransferred compound of hesperidin reached a peak 15 minutes after administration, and gradually decreased thereafter. On the other hand, hesperidin showed the same behavior as that of the hesperidin glycosyltransferred compound, but the blood concentration was about 1/20 or less of that of the hesperidin glycosyltransferred compound.

Example 2 A 5-week-old male rat (weight: about 300 g) preliminarily reared for 1 week was fasted all day and night.
Under anesthesia, 300 μl of a 5% (w / v) solution of hesperidin and a solution of hesperidin were directly administered to the duodenum. Controls received distilled water. After administration, blood was collected from the portal vein over time. The obtained blood was obtained from J.I.
A. After pretreatment according to the method of Boutin et al.
Hesperidin glycosyltransfer compounds and hesperidin contained therein were quantified using PLC. As a result, in the group to which the hesperidin glycosyltransferred compound was administered, the blood concentration reached 7.0 μg / ml 20 minutes after the administration, and thereafter gradually decreased. The blood concentration of the hesperidin glycosylated compound administration group was 5.5 μg / ml. At this time, hesperidin used for comparison was not detected in blood.

Example 3 A 5-week-old male Std. A ddY mouse (body weight, about 30 g) was fasted overnight and 300 μl of a 20% (w / v) glycosyltransferase compound of diosmin and a diosmin solution were directly administered to the stomach using a sonde. Controls received distilled water. After administration, blood was collected over time. The obtained blood was obtained from J.I. A. Bo
After pretreatment by the method of Utin et al., the diosmin glycosyltransfer compound and diosmin contained therein were quantified using HPLC.

As a result, the glycosyltransferred compound of diosmin was rapidly absorbed after administration, the blood concentration reached a maximum 15 minutes later, and was gradually metabolized thereafter. In contrast, diosmin was extremely poorly absorbed, and only a trace amount was detected in blood.

Example 4 A 5-week-old male Std. A ddY mouse (body weight, about 30 g) was fasted overnight, and a 20% (w / v) sugar-transferring compound of naringin and 300 μl of naringin solution were directly administered to the stomach using a probe. Controls received distilled water. After administration, blood was collected over time. The obtained blood was obtained from J.I. A. Bou
After pretreatment by the method of tin et al., the glycosyltransferred compound of naringin and naringin contained therein were quantified using HPLC.

As a result, the glycosyltransferred compound of diosmin was rapidly absorbed after administration, the blood concentration reached a maximum 15 minutes later, and was gradually metabolized thereafter. In contrast, naringin was extremely poorly absorbed, and only trace amounts were detected in blood.

Example 5 A 5-week-old male Std. A ddY mouse (body weight, about 30 g) was fasted overnight and 300 μl of a 20% (w / v) neohesperidin glycosyltransferase and neohesperidin solution was directly administered to the stomach using a probe. Controls received distilled water. After administration, blood was collected over time. The obtained blood was obtained from J.I.
A. After pretreatment according to the method of Boutin et al.
The glycosyltransferred compound of neohesperidin and neohesperidin contained therein were quantified using PLC. As a result, the glycosyltransferred compound of neohesperidin was rapidly absorbed after administration, the blood concentration reached a maximum 15 minutes later, and was gradually metabolized thereafter. On the other hand, neohesperidin was extremely hardly absorbed, and only a trace amount was detected in blood.

Example 6 SHR rats preliminarily reared on a normal solid diet for 1 week were fed with a diet of the same composition containing a 10% hesperidin glycosyltransfer compound and reared for 8 weeks. The rats in the control group were continuously fed a normal chow. The other test group was fed a feed of the same composition to which 10% hesperidin was added. Eight weeks later, when the blood pressure of the rats in each test group was measured, the blood pressure of the rats in the test group was about 5% lower than that in the control group, and the hypertension of the rats in the test group with the hesperidin-glycosylation compound was added. Symptom improvement became clear. Rats in the control group did not show any symptom-improving effect. In the hesperidin group, almost no improvement was observed. In addition, there was no significant difference in body weight gain or feed intake between the three test plots.

Example 7 An 8-week-old male Std. A ddY mouse (body weight, about 30 g) was fasted overnight, and a 4% (w / v) concentration of daidzin (daid) was used.
gin) (300 μl) or
A 4% (w / v) daidzin solution (300 μl) was administered directly to the stomach using a sonde. After administration, blood was collected over time. The obtained blood was obtained from J.I. A. After pretreatment according to the method of Boutin et al., The glucose transfer compound of daidzin and daidzin contained in blood were quantified using HPLC. As a result, as shown in FIG. 2, in the group to which the glycosyltransferred compound of daidzin was administered, the blood daidzin amount reached a peak 15 minutes after the administration. The amount of daidzin in blood referred to here is indicated by the sum total of daidzin and its glycoside, because a part of daidzyl transglycosylation compound was decomposed into daidzin by an enzyme in the living body. The concentration was about 6 times that of the daidzin-administered group.

Example 8 An 8-week-old male Std. ddY mice (body weight, about 30 g) were fasted overnight and genistin (gen) at a concentration of 4% (w / v).
istin) (300 μl) or a 4% (w / v) genistin solution (300 μl)
l) was administered directly to the stomach using a sonde. After administration, blood was collected over time. The obtained blood was obtained from J.I. A. After pretreatment by the method of Boutin et al., The glycin transfer compound of genistin and genistin contained in blood were quantified using HPLC. As a result, as shown in FIG. 3, in the group to which the genistin glycosyltransferred compound was administered, the blood genistin level reached a peak 15 minutes after the administration. The amount of genistin in blood referred to here is represented by the total sum of genistin and its glycoside, because a part of the glycin transfer compound of genistin was decomposed into genistin by an enzyme in the living body. The concentration was more than twice that of the genistin administration group.

Example 9 An 8-week-old male Std. A ddY mouse (body weight, about 30 g) was fasted overnight and a 10% (w / v) hesperidin glycosyltransferase solution (300 μl) or a 10% hesperidin solution (300 μl) was injected into the stomach using a probe. Was administered directly. After administration, blood was collected over time. The obtained blood was obtained from J.I. A. After pretreatment according to the method of Boutin et al., The glycosyltransferase compound of hesperidin and hesperidin contained therein were quantified using HPLC. As a result, the blood transglycosylation compound of hesperidin peaked at 15 minutes after administration, and gradually decreased thereafter. On the other hand, hesperidin showed the same behavior as the hesperidin glycosyltransferred compound, but the blood concentration was 1/20 or less of that of the hesperidin glycosyltransferred compound.

Example 10 A 20-week-old male rat (body weight, about 300 g) preliminarily bred for 1 week was fasted overnight, and under anesthesia, a concentration of 3% (w / v) hesperidin glycosyltransferase compound A solution (500 μl) or a 3% (w / v) hesperidin solution (500 μl) was directly administered to the duodenum. Controls received distilled water. After administration, blood was collected from the portal vein over time. The obtained blood was obtained from J.I.
A. After pretreatment according to the method of Boutin et al.
Hesperidin glycosyltransfer compounds and hesperidin contained therein were quantified using PLC. As a result, in the group to which the hesperidin glycosyltransferred compound was administered, the blood concentration reached 8.5 μg / ml 20 minutes after the administration, and gradually decreased thereafter. The blood concentration of the hesperidin administration group was 2.5 μg / m
l. At this time, in the water administration group used for comparison,
Hesperidin was not detected in blood.

Example 11 An 8-week-old male Std. A ddY mouse (body weight, about 30 g) was fasted all day and night, and a 2% (w / v) diosmin solution of a glycosyltransfer compound (300 μl) or a 2% concentration was used.
A (w / v) diosmin solution (300 μl) was directly administered to the stomach using a probe. After administration, blood was collected over time. The obtained blood was obtained from J.I. A. After pretreatment by the method of Boutin et al., The diosmin glycosyltransfer compound and diosmin contained therein were quantified using HPLC. As a result, the glycosyltransferred compound of diosmin is rapidly absorbed after administration, and the blood concentration becomes maximum after 15 minutes,
After that it was gradually metabolized. On the other hand, diosmin was extremely difficult to be absorbed, and only about 1/40 was detected in blood as compared with the group to which the glycosyltransferred compound of diosmin was administered.

Example 12 An 8-week-old male Std. A ddY mouse (body weight, about 30 g) was fasted overnight, and a 2% (w / v) solution of naringin in a glycosyltransfer compound (300 μl) or a 2% (w / v) concentration was used.
Naringin solution (300 μl) was administered directly to the stomach using a sonde. After administration, blood was collected over time. The obtained blood was obtained from J.I. A. After pretreatment according to the method of Boutin et al., The sugar transfer compound of naringin and naringin contained therein were quantified using HPLC. As a result, the glycosyltransferred compound of naringin was rapidly absorbed after administration, the blood concentration reached a maximum 15 minutes later, and was gradually metabolized thereafter. On the other hand, naringin was extremely hard to be absorbed, and its concentration was detected only about 1/30 as compared with the naringin glycosyltransferred compound administration group.

Example 13 An 8-week-old male Std. One ddY mouse (weight, about 30 g)
Fasted overnight, 300% of 2% (w / v) neohesperidin glycosyltransferase solution and 2% concentration
A neohesperidin (w / v) solution (300 μl) was directly administered to the stomach using a probe. After administration, blood was collected over time. The obtained blood was obtained from J.I. A. After pretreatment by the method of Boutin et al., The glycosyltransferase compound of neohesperidin and neohesperidin contained therein were quantified by HPLC. As a result, the glycosyltransferred compound of neohesperidin was rapidly absorbed after administration, the blood concentration reached a maximum 15 minutes later, and was gradually metabolized thereafter. On the other hand, neohesperidin is extremely difficult to be absorbed, and is about 1 times lower than that of the group to which neohesperidin is administered.
Only / 15 was detected.

[0028]

EFFECTS OF THE INVENTION A comparison between the case where a physiologically active flavonoid glycosyltransferred compound is orally administered and the case where a physiologically active flavonoid glycosyltransferred compound is orally administered improves the absorbability of the physiologically active flavonoid glycosylated compound into a living body, and the flavonoid's medicinal effects can be rapidly expressed. Was. In addition, since the absorbability was increased, a small dose was sufficient.

[0029]

[Brief description of the drawings]

FIG. 1 shows the time course of blood speridine concentration after hesperidin and hesperidin glycosyltransferred compound (unit of the vertical axis is mg / mg)
ml, the unit of the horizontal axis is time)

[Explanation of symbols]

White circles indicate hesperidin, and black circles indicate hesperidin glycosyltransferase.

[0030]

FIG. 2 shows the time course of blood daidzin concentration after administration of daidzin and daidzyl glycosyltransferase compound (units on the vertical axis are μg / ml,
The unit of the horizontal axis is time (minute))

[Explanation of symbols]

Black triangles indicate daidzin and black circles indicate daidzyl glycosylated.

[0031]

FIG. 3 shows the time course of blood genistin concentration after administration of genistin and a genistin glycosyltransferase compound (unit of the vertical axis is μg /
ml, the unit of the horizontal axis is time (minute))

[Explanation of symbols]

A black triangle indicates genistin, and a black circle indicates genistin glycosylated.

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Claims (2)

[Claims] Claims: 1. A food and drink and a medicine comprising a glycosylated compound of a bioactive flavonoid. 2. The food, drink and medicine according to claim 1, wherein the bioactive flavonoid is hesperidin, diosmin, naringin, neohesperidin, daidzin, genistin.