JPH01226824A - Metabolic promoter for urea nitrogen - Google Patents

Abstract

PURPOSE:To obtain the subject promoter effective in preventing or ameliorating renal function disorder, especially alcoholic urea nitrogen cacochymia, by using a flavonoid, saponin or their glycoside existing in a plant of family Legminaceae. CONSTITUTION:The objective agent contains at least one kind of compound selected from a flavonoid separated from a plant of family Legminaceae (e.g. arrowroot or mung bean), e.g. the isoflavonoid of formula I (R1-R6 are H, OH, OCH3, glucose, O-glucose or O-glucose xylose), a saponin, e.g. triterpenoidal saponin of formula II (R7 and R8 are H, CH3 or CH2OH) or their glycosides, as an active component. The compound of formula I is e.g. isolidone or genistein and the compound of formula II is e.g. 3-0-{alpha-L-rhamnopyranosyl(1 2)-beta-D- galactopyranosyl(1 2)-beta-glucuronopyranosyl}-soyasapogenol B.

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention relates to legumes, such as i (Puerarla
1obata Ohwl), mung bean (Phaseolu
The present invention relates to a urea nitrogen metabolism improving agent containing as an active ingredient at least one selected from the group consisting of flavonoids, saponins, and glycosides thereof obtained from Radiatus L.).

Fujie kudzu has long been used as kudzu starch as a raw material for Japanese sweets, and its roots and flowers are called kudzu root and 8 flowers, respectively, and have been added to Chinese herbal medicine formulations. In particular, kudzu root is supervised by the Ministry of Health and Welfare.
- According to the ``Guide to Prescribing Chinese Herbal Medicine'', it is included in 9 out of 210 prescriptions of Kampo medicine, and in the representative prescription Kakkonto, kudzu root is the main drug.On the other hand, 8 flowers are m-sake, It is prescribed and used as a Chinese herbal medicine in Kuzuhana Kaiseto, which aims to ``relieve alcohol poisoning.''

Kudzu plants contain flavonoids, especially infravonoids such as daidzein, genistein, and caccharide (Yakushukan 75
71959) (Chem, Pharm, Bull 1
3124491975) and saponins, especially triterpenoidal saponins (Che■, Phar-6Bu1)
1. 1293 (1985)) has already been found as its main component. Regarding medicinal effects, it has sweating, antipyretic effects (Japanese Pharmacological Journal only 263194+), and antispasmodic effects (Pharmacological Journal Sub-8B).
31959'), blood pressure lowering effect (Chinese Medical Journal Dan 265
1974), choleretic action (Applied Pharmacology 5 (2)
2471971 > In addition to the adrenergic beta receptor blocking effect (Kukugaku Gakuho 1fi 2181980), it also has a muscarinic effect on removed organs (Applied Pharmacology I 2471).
97+) and pavaberin-like effects (Chem, Pharm,
There are reports such as Bull 2317981975). In addition, reports related to the effects of sake include Tsukamoto, Kada, etc. (Proc, Symp, 1 Fakan-Yaku 15
123 +982) (Nihon University Medical Journal ff (8) 88
71983) has only reported that, regarding alcohol metabolism, infravon, a component of kudzu flowers, promotes the disappearance of acetaldehyde in the body. Considering the extremely wide range of ways in which it is used in Chinese medicine, it must be said that the elucidation of its medicinal effects is extremely insufficient.

The present inventors focused on the effects of refined sake, and determined the alcohol and acetaldehyde in the blood of the respective fractions containing flavonoids and saponins and their glycosides obtained from Kudzu root and Kazuhana. When examining the effect on the amount of It was found that the drug has preventive and ameliorative effects on disorders.

Isoflavonoids and triterbenoidal saponins in which the present inventors have found preventive and ameliorative effects on urea nitrogen metabolic disorders contain compounds represented by the following formulas -Funenin (I) and (II), respectively.

In the formula, R1, R2, R3, R4, R5, and R8 are the same or different and represent 810H% OMex glucose, 0-glucose, and 0-glucose xylose. Such flavonoids include
In particular, irisolidone, genistein, tectorizin, caccharidone, caccharin, caccharide, daidzein,
Infravons such as daidzin, puerarin, PG-1, PG-3, and formononetin are known, and all of them are compounds known in the literature.

That is, in the formula, R1 and R3 are OH groups, R2 and R6 are CH3
0 group, R4 and R5 are H, the compound is irisolidone, R1, R3 and R6 are OH groups, and R2, R4 and R5 are H
The compound is genistein, R1 and RB are OH groups,
R2 is CH30 group, R3 is O-glucose, R4 and R5
The compound in which is H is tectolizine, and R1 is an OH group, R
A compound in which 2 and R6 are CH30 groups, R3 is O-glucose, and R4 and R5 are H is caccharidone, and a compound in which R1, R4, and R5 are H1R2 and R6 is OH group, and R3 is CH30 group is caccharidone, and R1 is OH group, R2 and R6 are CH3
0 group, R3 is O-glucose-xylose (hereinafter, 〇-
gu1. -Xy1. ), the compound where R4 and R5 are H is cancalide, and the compound where R1, R2, R4 and R5 are H, R3
A compound in which R6 and R6 are OH groups is daidzein, and R1 and R
2, R4 and R5 are H, R3 is O-glucose, RG is O
The compound whose H group is daidzine, and RI, R2 and R5
is H, R3 and R6 are OH groups, and R4 is glucose. Buerarin is a compound in which R1 and R2 are H, R3 and R5
A compound in which R6 is an OH group and R4 is glucose is PG-1
, the compound in which R1 and R2 are H, R3 and R6 are OH groups, R4 is glucose, and R5 is CH30 group is PG-3, R1 and R
2, R4 and R5 are H, R3 is OH group, R11i is CH3
The compounds with the 0 group are each known as formononetin.

It is also represented by -shipman (n) or a CH20H- group. As triterpenoidal sabonin or its glycoside, 3-0-
(α-L-Rhamnopyranonol(1→2)-β-D-galactopyranosyl(1→2)-β-D-glucuronopyranosyl) Soyasapogenol B (In the formula has a CH20H group), 3-0-<α-L-
Rhamnopyranosyl (1 → 2)-β-D-galactopyranosyl (1 → 2)-β-D-glucuronopyranosyl) sophalanool (in formula 2, R7 is CH3 group, R8 is CH2
0H) A), 3-Oiα-L-rhamnobilanone(1→2)-β-L-arabinopyranosyl(1
→2) -β-D-glucuronopyranosyl) sofaradiol (in the formula (1), R7 means a CH3 group and R8 means a hydrogen group) is already known in the literature.

As is clear from the pharmacological tests described below, these ingredients are associated with experimental urea nitrogen metabolic disorders, especially alcoholic urea nitrogen.
It has preventive and improving effects on metabolic disorders.

By the way, urea is a substance produced from protein catabolism, and is a decomposition product produced in the process of detoxifying ammonia, which is highly toxic to living organisms. Its biosynthesis is carried out by the urea cycle in the mitochondria of the liver, and then
It is excreted in the kidneys as urine, and is the only compound used by mammals to excrete nitrogen derived from protein.

Blood urea nitrogen level is a comprehensive value related to diet, protein metabolism, and renal function. Fluctuations in this value can occur due to intake of large amounts of protein in the diet, extreme dehydration, anemia, blood for the gastrointestinal tract, or prolonged strenuous exercise. This occurs when proteins need to be mobilized and when some kind of abnormality occurs in renal function, and in this case, blood urea nitrogen increases. Among these, it is said that the most important factor is an increase in blood urea nitrogen caused by diseases directly or indirectly related to renal dysfunction.These diseases include nephritis, renal failure, uremia, Urinary tract obstruction, liver disease, high blood pressure, gout,
There are many diseases such as congestive heart disease, diabetes, and other infectious diseases, but there are still very few drugs that can prevent or improve the rise in blood urea nitrogen that occurs under these conditions. (For example, among the herbal medicines formulated with Chinese herbal medicine, rhubarb, huanglian, etc. have been recognized to have preventive and improving effects.)
Journal of the Japanese and Chinese Pharmaceutical Society, Vol. 14781981)
(Japanese and Chinese Pharmaceutical Society Journal, Vol.

Furthermore, among the pharmaceuticals actually used in clinical practice, there are no rapidly effective drugs for these diseases, and even now, the treatments for these diseases include a combination of bed rest, dietary therapy, and steroids. In this situation, where the application of (especially in the case of renal disease) is mainstream,
The present invention, which relates to a preventive and ameliorating agent for urea nitrogen metabolic disorders, which contains known compounds flavonoids and saponins contained in eight plants as essential active ingredients, has been developed by the present inventors. It is extremely significant for the treatment and prevention of the above-mentioned diseases.

The preventive and ameliorating agent for urea nitrogen metabolism disorder of the present invention is composed of an infravonoid or triterpenoidal saponin as an active ingredient, to which is added a solid or liquid pharmaceutically acceptable excipient. .

The dosage form for administration is usually powder, tablet, emulsion,
In addition to oral forms such as capsules, drugs, granules, and liquids (including alcoholic preparations, tinctures, preparations, liquid extracts, syrups, etc.), they may also be used internally as suppositories.

As solid or liquid excipients used here, those known in the art are used. To list some specific examples, powders and other powders for internal use include lactose, starch, dextrin, phosphoric acid, synthetic and natural aluminum silicate, magnesium oxide,
Examples of excipients in liquid formulations include dry aluminum hydroxide, magnesium stearate, and dried yeast.
water, chrycerin, simple syrup, propylene glycol,
Ethanol, fatty oil, ethylene glycol, polyethylene glycol, sorbitol, etc. are used, and the infravones and triterpenoidal saponins used as active ingredients in this invention are known compounds, and their production methods are also known. However, it can be produced, for example, by the following method.

The roots or flowers of the Chinese laurel plant are extracted with lower aliphatic alcohols or hydrous lower aliphatic alcohols, and the extract is concentrated under reduced pressure to obtain an extract. This was suspended in water, partitioned with the same amount of n-butanol, the dried butanol layer was degreased with hexane, and further n-butanol, ethyl acetate,
Distribution was performed with water, and isoflavonoids were obtained from the precipitate produced at this time (hereinafter, those obtained from arrowroot are referred to as PR-IF
(hereinafter abbreviated as PF-IF) obtained from 8 flowers.
, and the ethyl acetate and aqueous layers were Sephadex LH-20.
The triterpenoidal saponin was purified by the usual purification method using t'J (the product obtained from 8 flowers is hereinafter referred to as PF-SP).
Furthermore, after separating the aqueous layer obtained by partitioning water and n-butanol with Diaion chromatography, isoflavonoids can be isolated by repeated separations using Sephadex LH-20 column chromatography, etc. Obtainable. The physical properties of PF-IF, PR-IF and PF'-8P and the substances contained in these components are as follows.

First of all, PF-IF and PR-IF are brownish-white and white-white amorphous powders, respectively, and are tasteless or odorless powders with a slightly sweet taste, and both are easily soluble in methanol and dilute methanol.
Soluble in water and ethanol, sparingly soluble in chloroform, ether, hexane, and carbon tetrachloride. When measuring the UV absorption of these substances, a maximum absorption characteristic of isoflavonoids near 251 nm and a shoulder near 263-310 nm were observed. Furthermore, these substances were subjected to ordinary purification methods such as liquid chromatography, and the above (
Irisolidone, genistein, tectolidine, casokalydone, kanocathin, and cancalide were obtained as shown in formula I), and daidzein, daidzin, puerarin, PG-1, PG-3, and formononetin were obtained from PR-IF. These substances were confirmed by measuring their melting points, infrared absorption, thin layer chromatography, carbon and proton NMR, and other physical constants.

On the other hand, PF-8P is a yellowish-white-brown powder with a slight tinge and is odorless.It is easily soluble in methanol and diluted methanol, soluble in water and ethanol, chloroform, ether, etc.
Slightly soluble in hexane and carbon tetrachloride. Further, this substance
Perform ordinary purification methods such as liquid chromatography to obtain PF-
Three types of compounds shown in general formula (n) were obtained from 5P. All of these are white amorphous powders, and the physical constants of these compounds were measured or confirmed using the same procedure as above. Note that isoflavonoids and triterpenoidal saponins can also be obtained from mung beans by the above extraction method.

Reference examples of producing infravonoids and triterpenoidal saponins by the above method and the results of pharmacological experiments are shown below, but these examples explain the present invention in more detail, and the present invention is in no way dependent on these descriptions. It is not limited.

Reference Example 18 Production of Isoflavonoid and Triterpenoidal Saponins from Plants Eight flowers are roughly cut, and 1.85 kg thereof is extracted with 10-g of methanol under reflux at a temperature of GO'C. This operation was repeated 4 times to remove the solvent to obtain 632.8 g of methanol extract. Next, this extract was divided into n-butanol and 1 t of water, and 345.8 g of aqueous layer extract and 287.0 g of butanol layer extract were obtained. I got it. After defatting this butanol extract with hexane, it was distributed between n-butanol, ethyl acetate, and water (1:4:5). )23.
On the other hand, both fractions other than the precipitate, water, butanol, and ethyl acetate fractions were further purified using MCI gel CH
Using P20P, flow down with 10%-50% methanol,
After partitioning the first two parts obtained between chloroform and water, the aqueous layer was separated using Bondapak CI8, here 40% -
From the fraction obtained by flowing down with 70% methanol, triterbenoidal saponin (PF-8P) 5 is extracted according to a conventional method.
0.0g was able to be obtained.

In addition, P can also be obtained from 345.8 g of the aqueous layer extract by performing conventional minute purification methods such as diamond ion chromatography and Sephadex column chromatography.
It is also possible to obtain F-IF.

In history, kudzu root 3. Butanol layer extract 63. OKg is coarsely cut and processed according to the above-mentioned 8 flower separation method.
From 0 g, 47.0 g of infravonoids (
PR-IF) and triterbenoidal saponin3. Og
I was able to say wo (J).

Next, the inhibitory effect of the drug according to the present invention on blood urea nitrogen metabolic disorder will be shown.

Experimental Example 1 1.0 to 7 pairs of heavy ddV male mice weighing 25-35 g.
5. The test drug suspended in gum arabic was administered orally, and 1 hour later, the test drug was administered at a concentration of 50% (W/V) 1/-. Og/Kg was administered orally, and 4 hours after this, blood was collected from the heart through chest incision, serum was separated by centrifugation, and the urea nitrogen (BUN) value in this serum was measured using the urease indophenol method. did.

The normal group referred to here refers to a group in which only gum arabic solution was administered instead of the test drug, and alcohol was not administered.
Furthermore, the control group referred to herein refers to a group in which gum arabic was administered instead of the test drug, and then 5.0 g/Kg of the above-mentioned alcohol was administered. Furthermore, as a comparative test, we also conducted a comparative test using a large amount that is clinically said to be effective for urea nitrogen metabolic disorders.

The results are shown in Table 1. Note that the inhibition rate (%) was calculated using the formula F.

From these results, we can determine the tendency of alcohol-induced increase in serum urea nitrogen levels to be
In all cases, it was found that the inhibition rate was approximately 60-70%, which was statistically significant.

Furthermore, it was confirmed that the effect was far superior to that of the large-scale mass shooting (Shinshu hostages) used as a comparison sample.

The mechanism by which blood urea nitrogen increases due to alcohol administration is said to be caused by renal dysfunction due to a decrease in renal glomerular filtration rate (J, 5tud, Alc, 19203-
2251958) (Jpn, J.

Alcohol & Drug Dependence
17 (4) 327-3401982) Therefore, the fact that flavonoids and saponins suppressed the increase in blood urea nitrogen indicates that they ameliorate the decrease in renal glomerular filtration rate caused by alcohol, and suggest that flavonoids and saponins suppress the decrease in renal glomerular filtration rate. This gives us hope for an improvement effect.

Table 1 * indicates significance compared to the control group at p<0.05.

(Test) Next, an acute toxicity test for oral administration of PF-IF, PR-IF, and PF-8P was conducted using ddY-strain male mice with 5 mice per group. As a result, all three drugs were 50%
No deaths were observed within 72 hours at 00+g/Kg X. These results demonstrate that the drug of the present invention is an agent for preventing and ameliorating urea nitrogen metabolic disorder with extremely low toxicity and high safety when considering the effective dose.

Furthermore, the effective dose for humans has been determined based on previous studies and a large number of clinical curative doses (Japanese and Chinese Pharmaceutical Society Journal Vol.
1G221983), it is usually appropriate for adults to take 350-700 mg orally in 113-4 divided doses for all three drugs.

Example 1 a) Preparation of granules 30 g of PF-IF or PR-IF or PF-SP is made into a fine powder, and 119 g of lactose and 1 g of magnesium stearate are mixed therein, and this mixture is compressed into tablets with a diameter of 20 Il. Take Slanog tablets. This is crushed, sized,
The mixture was sieved to obtain 20-50 mesh granules.

In addition, this granule is administered once depending on the symptoms.
200mg can be taken 3-4 times a day.

b) Production method of capsules 125 mg of the above granules or fine powder thereof
was filled into No. 3 gelatin capsules to obtain capsules.

In addition, this capsule is administered once per dose depending on the symptoms.
0 capsules, can be taken up to 3 times a day.

Example 2 25g of PF-IF or PR-IF or PF-8P was made into fine powder, and each powder was mixed with 73g of lactose, 22g of microcrystalline cellulose.
0 g and 7 g of magnesium stearate, and this mixture was compressed into tablets using a single-shot tablet machine to form tablets with a diameter of 7 cm and a weight of 1.
25 mg tablets were manufactured.

In addition, this tablet contains 25++ of each of the above drugs in one tablet.
+g What does it contain? This tablet can be taken 5-10 tablets at a time, 3-4 times a day.

Claims (6)

[Claims] (1) A urea nitrogen metabolism improving agent containing as an active ingredient at least one selected from the group consisting of flavonoids, saponins, or glycosides thereof contained in leguminous plants. (2) The urea nitrogen metabolism improving agent according to claim 1, wherein the flavonoids, saponins, or their glycosides are contained in kudzu plants. (3) Flavonoids or their glycosides have the general formula (I) ▲
There are mathematical formulas, chemical formulas, tables, etc. ▼ (I) In the formula, R1, R2, R3, R4, R5, R6 are the same or different and mean H, OH, OMe, glucose, O-glucose, O-glucose xylose. The urea nitrogen metabolism improving agent according to claim 1 or 2, which is a compound represented by the following. (4) Flavonoids are isolidone, genistein, tectolizine, caccharidone, caccharin, caccharide,
daidzein, daidzin, puerarin, PG-1, PG-
3. The urea nitrogen metabolism improving agent according to claim 3, which is an isoflavone called formononetin. (5) Saponin has the following general formula (II) ▲ There are mathematical formulas, chemical formulas, tables, etc. ▼ (II) In the formula, R7 and R8 are the same or different, H, CH3-
The urea nitrogen metabolism improving agent according to claims 1 and 2, which is a triterpenoidal saponin or a glycoside thereof represented by , or CH2OH- group. (6) Saponin is 3-O-{α-L-rhamnopyranosyl(1→2)-β-D-galactopyranosyl(1→2)
-β-D-glucuronopyranosyl} Soyasapogenol B, 3-O-{α-L-rhamnopyranosyl (1→2)-
β-D-galactopyranosyl (1→2)-β-D-glucuronopyranosyl}soradiol, 3-O-{α-
L-rhamnopyranosyl (1→2)-β-L-arabinopyranosyl (1→2)-β-D-glucuronopyranosyl}
The urea nitrogen metabolism improving agent according to claim 5, which is sofaradiol. 7) Claim 1 for oral or parenteral administration
An agent for preventing and improving urea nitrogen metabolic dysfunction (renal dysfunction) according to paragraphs 6 to 6.