JPS638328A - Remedy for liver disease - Google Patents

Abstract

PURPOSE:A remedy for liver disease having high treating effect and low toxicity, comprising ajoene as an active ingredient. CONSTITUTION:A remedy for liver disease containing ajoene shown by the formula as an active ingredient. A dose is 10mg-10g/person/day, preferably 50mg-5g/person/day. The compound shown by the formula as the active ingredient is obtained by grinding (e.g. Allium sativum L. form a pekinese Makino, etc.; Allium sativum L. belonging to the genus Allium of Liliaceae) to carry out reaction readily, adding the ground garlic to a hydrous lower aliphatic alcohol adjusted to pH 2-6 with an organic or inorganic acid, effecting reaction at room temperature or under heating, extracting the prepared reaction solution with an organic solvent such as chloroform, etc., and carrying out normal phase chromatography or reversed phase chromatography.

Description

DETAILED DESCRIPTION OF THE INVENTION BACKGROUND OF THE INVENTION Technical Field The present invention relates to novel uses of AjOene. The present invention relates to a therapeutic agent for liver diseases containing ajoene as an active ingredient.

Prior Art Attempts to search for medicinal substances useful for the human body from natural products have been carried out since ancient times, and it is well known that many useful substances are available as medicines.

For example, garlic, which is generally known as a tonic, has been used since ancient times as a stomachic, diaphoretic, diuretic, expectorant, intestinal regulation, sterilizer, and anthelmintic. In recent years, the platelet aggregation inhibiting effect of garlic has attracted attention, and it has been revealed that methylallyl trisulfide present in the volatile components of garlic (so-called garlic oil) has this activity (Lancetll (8212), 150). -1
51 (1982)), it has also been reported that allicin in garlic has similar activity (Phys.
tochemistry, 24.1593-1594
(1985)). In addition, Block
) isolated the sulfur-containing compound ° (ajoene) shown by the following structural formula (I) from garlic, and reported that it has the effect of inhibiting platelet aggregation (J, Am
er, Chem, Soc, , 106.8295~
8296 (1984)).

<I> In addition, the platelet aggregation inhibitory action mechanism d4 of ajoene is 1! from the previously reported Allium genus!
It has been shown that the mechanism of action of the above-mentioned compounds (methylallyl trisulfide and allicin) is different from that of the previously described compounds (Thrombosis Research, 3).
2.155-169 (1983)). Furthermore, currently
Ajoene is expected to be applied in the cardiovascular field and other areas such as cerebral thrombosis and arteriosclerosis.

On the other hand, Japan is currently transitioning into an aging society due to improved living standards and advances in medical technology, and adult diseases (medically speaking, diseases that occur after the age of 40) are becoming a major social problem. It has become.

One of these adult diseases is liver disease, which is the largest organ in the human body and is an important organ responsible for various substance metabolism, detoxification, etc., and compared among all adult diseases. This accounted for a high proportion of the population. Liver diseases are mainly caused by viral hepatitis, liver cell damage caused by drugs (anti-tuberculosis drugs, antibiotics, sulfa drugs, etc.), and liver cirrhosis caused by alcoholic beverages.There are various treatments for these liver diseases. Medications are used, such as hormones (insulin, etc.), vitamins (vitamin B1, vitamin E, etc.)
, pan-l-thenic acid, etc.) and other drugs (methionine, orotic acid, brocuronic acid, glycyrrhizin, etc.) are used as appropriate depending on the cause and symptoms. However, among the above drugs, hormonal drugs have problems such as side effects and relapse of symptoms when administration is stopped.
In addition, where vitamin supplements need to be administered by hanging from the ceiling,
Water-soluble vitamins are ineffective when liver damage is significant, and fat-soluble vitamins may cause overdose.
There were various problems such as the lli case.

[Summary of the invention]

With the aim of solving the above problem, the present inventors have conducted extensive research and have newly discovered that ajoene, a sulfur-containing compound represented by the above structural formula (I), has a therapeutic effect on liver diseases. As a result, a therapeutic agent for liver diseases according to the present invention has been completed.

Therefore, the therapeutic agent for liver diseases according to the present invention is characterized by containing ajoene as an active ingredient.

The therapeutic agent for liver diseases according to the present invention contains the above-mentioned ingredients as active ingredients, and the fact that the above-mentioned ingredients have a therapeutic effect on liver diseases is an unexpected finding even for those skilled in the art, and the above-mentioned problems can be solved. In addition to solving the problem, it also has the following advantages.

1) Active ingredient of the drug for treating liver disease of the present invention which has a large therapeutic effect on liver disease (- Active ingredient of a drug conventionally used as a therapeutic agent for liver disease [-- Smaller amount than cysteine, but equivalent therapeutic effect on liver disease) (See experimental examples below).

11) As mentioned above, it is known that the active ingredient of the liver disease therapeutic agent of the present invention, which has low toxicity, is contained in garlic. It goes without saying that garlic is commonly used as food and that a dilute ethanol extract of garlic (which will be described later) contains the active ingredients of the drug of the present invention.

), the D5o value is the maximum physiologically administrable dose (4,5!7
/Ng) and from the experimental examples in mice mentioned above, it can be said that the liver disease therapeutic agent of the present invention has low ib properties. In addition, it was reported that no abnormalities were observed in physiological or pathological tests after long-term (6 months) oral administration (2 g/Kg) of garlic extract (above) (J ,Toxicol,Sci,, 9.61
(1984)], its side effects are inconceivable.

As described above, the active ingredient of the liver disease therapeutic agent of the present invention has no side effects and is low in efficacy as described above, and has a higher efficacy than conventional drugs. It is expected that it will make a great contribution in the medical field of drugs for treating chronic diseases, which contain the F component as a 9j component.

[Detailed description of the invention]

Ajoene and its acquisition Ajoene can be extracted and isolated from garlic, for example, as described above. More preferably, it can be obtained by the method described in Japanese Patent Application No. 60-268177, that is, the method specifically shown below. However, in this case, strictly speaking,
Rather than simply subjecting garlic to extraction to extract the ajoene in garlic, ajoene is produced from an ajoene precursor compound.

(1) Raw material garlic The garlic used here is from the Liliaceae family.
ae), Allium sativum Linnaeus belonging to the genus Allium
m L, ), for example, giant garlic (Alli
um sativum L.

Forma pekinese Hakino) corresponds to this.

The part to be used as the material for obtaining the desired fraction is particularly preferably the bulb part (which usually contains 5 to 20 small blastomere-shaped bulbs that have been split internally), which can be dried or It can be used for extraction as is.

(2) Processing method of garlic Garlic is crushed to be easily reacted, preferably homogenized, and then adjusted to pH 2 using any pharmaceutically acceptable organic or inorganic acid such as phosphoric acid, hydrochloric acid, acetic acid, citric acid or tartaric acid. ~6 (more preferably pH 3 to 4) of water containing about 1 to 4 carbon atoms, preferably 1 to 2,
A lower aliphatic alcohol (usually a monohydric alcohol) is added to the reaction mixture at room temperature or under heating.

Processing of garlic with lower aliphatic alcohols is carried out under hydrous conditions. Here, when referring to the water-containing condition, water derived from garlic is also taken into consideration. Therefore, when raw garlic is used, it is considered to be processed under hydrated conditions even when absolute alcohol is used. In this sense, the "water-containing condition" is preferably an alcohol concentration of about 5 to 95%, preferably about 20 to 90%.

Under such acidic and water-containing conditions, the reaction time is long, half a day to two days (preferably about one day) at room temperature, and several minutes to several tens of hours under heating (preferably below the boiling point of the system). The reaction time is usually short (preferably about 30 minutes to 10 hours at 10 to 90°C). If necessary, a coenzyme of allinase, such as pyridoxine phosphate or pyridoxine hydrochloride, may be added in advance when crushing or homogenizing garlic.

The thus-depleted reaction solution is concentrated under reduced pressure either as it is or after filtration, extracted with any organic solvent such as chloroform, ethyl acetate, etc., and extracted with I'? The target ajoene can be obtained by subjecting the extracted extract to normal phase chromatography, reversed phase chromatography, or a combination thereof.

In addition, the residue obtained after filtration can be extracted 2 to 3 times with any organic solvent, such as chloroform, ethyl acetate, lower alcohol (1fiI alcohol having 1 to 3 carbon atoms [preferably alcohol]), or acetone. Ajoene can be obtained by collecting the ajoene remaining in the residue and performing chromatography in the same manner as above.

(3) Ajoene Ajoene as used in the present invention refers to an oil-like substance obtained from garlic, for example, and refers to a compound represented by the above structural formula (I). This compound has geometric isomers of E-ajoene and Z-ajoene, both of which are included in the present invention.

Liver disease therapeutic agent The liver disease therapeutic agent according to the present invention contains ajoene represented by the above formula (I) as an active ingredient. This drug consists of ajoene alone or mixed with auxiliary ingredients in liquid or solid preparations, such as excipients, binders, diluents, etc., and is available in powders, granules, capsules, injections, It can be administered orally or parenterally in any dosage form such as a solution. Further, other drugs may be mixed as necessary. Dosage depends on age, weight,
The dose may be increased or decreased as appropriate depending on the symptoms, but when administered orally or by static infusion, the dose is 10 mg to 10 g per day for adults, preferably 50 g.
mr+=5q, approximately. Another preferred embodiment of the present invention is an intermediate dosage form in which the above-mentioned daily dosage is administered in one or several divided doses.

In addition, five 5-week-old ICR male mice were used, and ajoene was administered at 3001n! Administered intraperitoneally at a ratio of 7/k (+), 1
As a result of weekly observation, no deaths were observed.
Ajoene can be said to have low toxicity.

Therapeutic action for liver diseases The therapeutic action for liver diseases as used in the present invention refers to the improvement of various pathological conditions caused by degenerative necrosis of hepatic parenchymal cells caused by internal and external factors.

In the present invention, confirmation of the above-mentioned effects was conducted using carbon tetrachloride (this substance destroys hepatocytes and causes glutamate oxaloacetate transferase (GOT) to Tl121) and the active ingredient of the liver disease therapeutic agent of the present invention in experimental animal mice. After administering G.
Measure OT (J, clin.

Invest, 34.126 (1955), Clinical Pathology, 26.70 (1978)).

EXPERIMENTAL EXAMPLE According to the present invention, the liver therapeutic action of the hepatolytic drug was investigated using its active ingredient ajoene in the following intracellular carbon tetrachloride liver injury model.

1) After preliminarily breeding ddY male mice for the first time in the experiment (6-week-old mice purchased from Shizuoka Dokyo) for one week, mice that appeared to be healthy (
(Weight: 29 to 31 g) were selected and used in the following experiments. The mice were kept at a room temperature of 24±1°C and a humidity of 55±5°C.
%, ventilation frequency 15 times/hour, 15 hours of lighting, 10 hours of lighting, and five mice each were placed in flat-bottom cages [
The mice were housed in Nippon Yurea Co., Ltd. food (GE-2 (Nippon Yurea Co., Ltd.) A3 and distilled water were freely available to the mice.

2) The drug ajoene was obtained by the method described in Reference Example below.

Since ajoene is fat-soluble (it dissolves in ethanol but is stable in alcohol), it was diluted with distilled water and used.

The final ethanol concentration was 12%.

In addition, it is an active ingredient in currently used liver disease treatments.
Cysteine was used as an indicator of the efficacy of the drug of the present invention as a therapeutic agent for liver diseases. Here, L-cysteine dissolved in a 1% aqueous gum arabic solution was used.

CCl4 (Hakai Chemical) is added to olive oil (Yamazen Pharmaceutical) by 3
A solution dissolved at a ratio of %V/V was used.

3) Dosage The experimental animals were divided into 7 groups with 5 animals per group, and the following drugs were administered to each group.

1: 12% ethanol (control) 2: Ajoene 1η/)C9 3: Ajoene 10*//(g 4: Ajoene 100■/Ky 5: [-Cystine 240■/Kg6: Shi-Cystine 480Rg/N97: Arabia 1% aqueous solution of gum (control) where 1 ethanol and 7 gum arabic solutions were used as solvents for ajoene and L-cysteine, respectively.

4) Administration method The amount of administered liquid is 10 mg/K per animal for each CY.
Weight was expressed as g.

Drug administration was performed as follows. That is, oral administration was performed once a day for 2 days, followed by fasting for 18 hours after the final administration (i.e., the second administration, 24 hours after the first administration), and 1 hour after the final administration. Then CCl4<
2 mg/9 body weight) was administered subcutaneously.

After administering CCl4, feed, administer cc, 4, then 2
After 4 hours, blood was collected according to the local authority, and 1 j of serum was added according to the usual method.
is.

5) Measurement of serum GOT value Serum GOT value is measured by transaminase C test Wak.
It was measured by e (Wako Suisyaku). Note that the test solution was appropriately diluted with distilled water and then immediately measured.

6) Results The serum GOT values were as shown in Table 1. In addition, the numerical values in the table indicate the standard error of the mean, and all are the results when five animals were used.

As a result, ajoene had no effect at low doses, but at medium doses (9 in 10η) and high doses of ff1 (9 in 100).
■/Kg) showed an inhibitory effect. Both the middle and high dose groups showed similar levels of suppressive effects.

Ajoene is produced in the decomposition pathway of allicin,
Two molecules of ajoene are produced by combining three molecules of allicin. The former is very unstable, while the latter is very stable. Ajoene is known to have an anticoagulant effect, and this experiment showed that the active ingredient of this drug can cause liver damage equivalent to L-cysteine at a lower dose than L-cysteine, the active ingredient of currently used liver disease treatments. It was shown to have a protective effect.

Refer to Table 1 Example 1 (obtaining ajoene) Add 2 mg of pyridoxine hydrochloride to 170 g of garlic,
Homogenized. This was made up to a total volume of 350ml with water,
The pH was adjusted to 3-5 by adding citric acid. 350 for this
1 of ethanol was added and heated at 65°C for 4 hours. The reaction mixture was filtered and the solution was about 7,001 to about 2,001
It was concentrated under reduced pressure to ml and extracted twice with ethyl acetate 2001, and the ethyl acetate extract layer was concentrated under reduced pressure.

The obtained extract was subjected to silica gel column chromatography (developing solvent: ethyl acetate: acetone = 20:1) to yield 45.5 m of (E)-ajoene.

I got it.

2 (Ajoene@11) 3 mg of pyridoxine hydrochloride was added to garlic 200Q and homogenized. Make this all ff14001 with water,
The pH was adjusted to 3-4 by adding citric acid.

400ml of ethanol was added to this and heated at 65°C for 4 hours. The reaction mixture was concentrated to about half its volume under reduced pressure, extracted twice with ethyl acetate 4001, and the ethyl acetate layer was concentrated under reduced pressure.

The obtained extract was subjected to high performance liquid chromatography (column: TSK get 0DS120A (Toyo Kaidat Kogyo), eluent: 40% acetonitrile), and then subjected to (E
)-Ajoene61. I got Omg.

Note that the 13C NMR and MS spectrum data of ajoene obtained in this example and the above example both have the following values, and are in accordance with the literature values (J, Am.

Chew, Soc, 106.8295 (1984)
), it was confirmed and identified.

(E)-Ajoene FD-MS: m/z 234 (M+) 54.5. 116.9. 119.3. 123.7. 125.7. 132.6. 134.7

Claims (1)

[Claims] A liver disease treatment agent characterized by containing ajoene as an active ingredient.