JP3075358B2 - Liver function improver - Google Patents

Description

Description: TECHNICAL FIELD The present invention relates to a liver function improving agent containing a dioxabicyclo [3.3.0] octane derivative as an active ingredient.

[Conventional technology]

The liver, along with the brain, is the largest parenchymal organ in the human body, and has detoxification, carbohydrate metabolism, protein metabolism, bile production and secretion, blood coagulation factor production, hormone regulation, fat /
It has various functions such as storage of various biological constituents such as glycogen, protein, and vitamin. However, these functions are also acutely or chronically impaired due to viruses, drugs, toxic substances, excessive intake of alcohol, malnutrition, hepatic circulatory system disorders, bile duct obstruction, etc., and viral hepatitis,
It manifests as a drug addictive hepatitis, alcoholic hepatitis, congestive hepatitis, liver damage due to bile umbilical cord, fatty liver, jaundice, or eventually cirrhosis.

For example, in the case of alcoholic liver disease, treatment includes dietary fat intake and control of excessive alcohol intake, as well as antihistamines, barbiturates,
Adenosine triphosphate (ATP), pyrazole,
Amino acid preparations such as a mixture of dihydroxyacetone and riboflavin, glucuronic acid, arginine hydrochloride or glutathione have been used for drug therapy, but the effects of the drug are still unsatisfactory. At present there is no way. In the case of liver damage caused by drugs and poisons, one of the causes is the generation of harmful oxygen free radicals. Therefore, poisoning due to such drugs and poisons,
A large amount of glutathione is used for treating allergic diseases and the like. Glutathione is particularly effective in cell protection.
Reducing reactive oxygen species and free radicals by glutathione peroxidase to eliminate them or glutathione-S
-It reacts with poisons by transferase and exerts its action by excreting it out of cells as glutathione conjugate, and plays a role in anti-oxidation and detoxification as well as radiation damage protection. However, even if glutathione itself is administered, the half-life in blood is as short as several minutes and is not very effective in increasing tissue glutathione, and at present, glutathione derivatives and the like are being developed.

Therefore, development of a novel liver function improving agent is strongly desired.

[Problems to be solved by the invention]

Therefore, an object of the present invention is to provide a novel liver function improving agent.

[Means for solving the problem]

In order to achieve the above object, the present inventors considered that liver-derived enzyme activities (glutamate oxaloacetate aminotransferase (GOT), glutamate pyruvate aminotransferase (GPT)) in mouse and rat sera were improved. GOT, GOT, as a result of searching for liver function improving agents using GPT as an index, dioxabicyclo isolated from sesame seeds, sesame meal and sesame oil or obtained synthetically (3.3.0)
The present inventors have found that an octane derivative has liver function improving ability and high safety, and completed the present invention.

Accordingly, the present invention provides the following general formula (I): (Wherein R ¹ , R ² , R ³ , R ⁴ , R ⁵ , and R ⁶ are each independently a hydrogen atom, an alkyl group having 1 to 3 carbon atoms, or R ¹ and R ² , and / or R ⁴ And R ⁵ together represent a methylene group or an ethylene group, and n, m, l represent 0 or 1), a dioxabicyclo [3.3.0] octane derivative (all possible stereoisomers) Is included in the above structure definition)).

[Specific explanation]

The dioxabicyclo [3.3.0] octane derivative used in the present invention includes sesamin, sesaminol, episesamin, episesaminol, sesamolin, 2- (3,4-methylenedioxyphenyl) -6- (3-methoxy -4-
(Hydroxyphenyl) -3,7-dioxabicyclo [3.3.
0] octane, 2,6-bis- (3-methoxy-4-hydroxyphenyl) -3,7-dioxabicyclo [3.3.0] octane, or 2- (3,4-methylenedioxyphenyl)-
6- (3-methoxy-4-hydroxyphenoxy) -3,
7-dioxabicyclo [3.3.0] octane and the like, and these stereoisomers or racemates can be used alone or as a mixture.

The method of obtaining the compound of the present invention and an extract containing the compound of the present invention as a main component can be performed by the following procedure. First,
To obtain an extract containing the compound of the present invention as a main component from sesame oil, extraction and concentration using various organic solvents that are substantially immiscible with sesame oil and that can extract and dissolve the compound of the present invention. It is obtained by doing. Examples of such an organic solvent include acetone, methyl ethyl ketone, diethyl ketone, methanol, and ethanol. To obtain an extract containing the compound of the present invention as a main component,
For example, it is obtained by uniformly mixing sesame oil and any of the above-mentioned solvents, leaving the mixture to stand at a low temperature, performing phase separation according to a conventional method such as centrifugation, and evaporating and removing the solvent from the solvent fraction. More specifically, sesame oil 2-10 times,
It is preferably dissolved in 6 to 8 times the volume of acetone, and left at -80 ° C overnight. As a result, the oil component precipitates, and the organic solvent is distilled off from the filtrate obtained by filtration to obtain an extract containing the compound of the present invention as a main component. Alternatively, it can be obtained by mixing sesame oil with hot methanol or hot ethanol, leaving the mixture at room temperature, and evaporating and removing the solvent from the solvent fraction. More specifically, sesame oil is mixed with 2 to 10 times, preferably 5 to 7 times the volume of hot ethanol (50 ° C. or higher) or hot ethanol (50 ° C. or higher) and vigorously extracted. Phase separation is carried out according to a conventional method such as standing at room temperature or centrifugation, and the solvent is distilled off from the solvent fraction to obtain an extract containing the compound of the present invention as a main component. Supercritical gas extraction can also be used. To obtain each of the compounds of the present invention from this extract, the extract is treated according to a conventional method such as column chromatography, high performance liquid chromatography, recrystallization, distillation, liquid-liquid AC distribution chromatography, and the like. May be isolated. More specifically, a reversed phase column (5C _18), methanol / water eluent (60:40)
The above extract was fractionated by high performance liquid chromatography, the solvent was distilled off, and the obtained crystals were recrystallized with ethanol to give sesamin, episesamin, sesaminol, episesaminol, etc. The compound of the present invention is obtained. The sesame oil to be used may be a purified product or any crude product before the decolorizing step in the sesame oil production process. In addition, sesame seed or sesame seeds (defatted sesame seed, residual oil content of 8 to 10
%). In this case, after crushing sesame seeds or sesame seeds as necessary, extraction can be carried out by a conventional method using any solvent, for example, the solvent described above for extraction from sesame oil. After the extraction residue is separated, the extract is obtained by removing the solvent from the extract by evaporation or the like. From the sesame seed extract, sesame cake extract or crude sesame oil extract thus prepared, in addition to sesamin, episesamin, sesaminol, episesaminol,
Sesamoline, 2- (3,4-methylenedioxyphenyl)
-6- (3-methoxy-4-hydroxyphenyl) -3,
7-dioxabicyclo [3.3.0] octane, 2,6-bis-
(3-methoxy-4-hydroxyphenyl) -3,7-dioxabicyclo [3.3.0] octane or 2- (3,4-methylenedioxyphenyl) -6- (3-methoxy-4-
(Hydroxyphenoxy) -3,7-dioxabicyclo [3.
3.0] Octanes of the invention can be obtained in a similar manner. Further, the compound of the present invention can be obtained from a by-product of the sesame oil production process.

The method for purifying the compound of the present invention and the method for obtaining an extract are not limited thereto. Further, the compound of the present invention and the extract containing the compound of the present invention as a main component are not limited to those obtained from sesame oil, sesame cake, and sesame seed, and all natural products containing the compound of the present invention can be used. It is obvious, for example, Goka bark, Kiriki, White bark, Hichi , Fine spices and the like.

The method of obtaining the compound of the present invention by synthesis includes:
The following are mentioned.

For example, for sesamin and episesamin,
78 , 1242 (1956)], and pinoresinol (in general formula I).
R ₁ = R ₄ = H, R ₂ = R ₅ = CH ₃ , n = m = 1 = 0) is Freundenb
Syringaresinol (R ₁ = R ₄ = H, R ² = R ^{3 in} general formula I) according to the method of erg et al. [Chem. Ber., 86 , 1157 (1953)].
⁼ R ⁵ = R ^{6 =} CH ₃ , n = 0, m = 1 = 1) can be synthesized by the method of Freundenberg et al. [Chem. Ber., 88 , 16 (1955)].

Further, the compound of the present invention can be used in the form of a glycoside, and these can be used alone or in an appropriate combination as a liver function agent.

The liver function improving agent of the present invention can be orally administered,
Alternatively, it can be administered by parenteral administration, for example, intramuscular injection, subcutaneous injection, intravenous injection and the like.

The dose varies depending on the purpose of administration, the condition of the subject, and the like, but is generally 1 to 100 mg / day for oral administration and 0.1 to 20 mg / day for parenteral administration. For example, when preparing an injection, a solubilizing agent for pharmaceuticals, for example, a nonionic surfactant or the like can be used. More specifically,
The compound of the present invention can be prepared by heating and dissolving the compound of the present invention in a nonionic surfactant such as POE (60) hydrogenated castor oil or POE sorbitan monooleate in an 80-fold volume and diluting it with a physiological saline solution. Agents, stabilizers, preservatives, and soothing agents may be added. Furthermore, emulsion preparations, capsules, powders, granules, tablets and the like can be prepared as required.

Since the compound of the present invention is a compound found in conventional foods or a compound thereof, it is clear that the compound is also excellent in safety. This is also evident from the fact that no abnormal symptoms were observed when 2.14 g / day / kg of sesamin was continuously administered (orally administered) to 7-week-old ICR male mice for 2 weeks.

Next, the present invention will be described more specifically with reference to examples.

Example 1 To 16.5 kg of sesame oil, 94.5 of hot methanol (60 ° C) was added and mixed vigorously for extraction. The mixture was allowed to stand at room temperature overnight, and the organic solvent was distilled off from the upper layer of methanol with a rotary evaporator to obtain 424 g of an extract containing the present compound as a main component. Add acetone 3.2 to this extract and dissolve,
And left overnight. The organic solvent was distilled off from the filtrate obtained by filtration to obtain 103 g of an extract containing the present compound as a main component. As a result of analyzing the compound of the present invention in the extract, sesamin was 19.6%, episesamin was 30.6%, sesaminol and episesaminol was 10.2%, and the content of the compound of the present invention in the extract was 60.4%.

Eight-week-old males (CD-1 mice (Charles River 1 Japan) were bred for 1 week on a normal diet (commercially available CE-2 diet, CLEA Japan), and the mice were divided into four groups with 10 mice per group. One group was raised on a similar diet and the remaining three groups contained 1% cholesterol, 0.2%
They were fed on a high cholesterol diet supplemented with 5% cholic acid and 5% olive oil. Two of the three groups fed the high cholesterol diet were fed a diet to which the above-mentioned extract containing the present compound as a main component was further added at 0.4% and 0.6%. After breeding for 2 weeks, the animals were fasted for 15 hours and blood was collected. The GOT and GPT activities in the serum were analyzed using an automatic biochemical analyzer (Hitachi 7050). Table 1 shows the results. The increase of GOT and GPT caused by the high cholesterol diet could be respectively reduced by giving the compound of the present invention.

Example 2 After breeding on a normal diet for one week in the same manner as in Example 1, the mice were divided into four groups with 10 mice per group. One group was fed a similar diet and the remaining three groups contained 1% cholesterol, 0.2% cholic acid and 5%
They were raised on a high cholesterol diet supplemented with evening primrose oil. Two of the three groups fed the high cholesterol diet further received a purified mixture of sesamin and episesamin (sesamin: 61.5%, episesamin: 38.0%).
Feeds supplemented with 4% and 0.6% were given. After bred for 2 weeks, 1
Blood was collected after a 5-hour fast, and GOT and GPT activities in the serum were measured. Table 2 shows the results. The elevation of GOT by a high cholesterol diet could be reduced by giving a sesamin and episesamin mixture.

Example 3 Eight-week-old male SD rats (CLEA Japan) were bred on a normal diet for one week, and the rats were divided into four groups, each comprising six rats. One group is bred on the same normal diet,
The remaining three groups included 20% casein, 10% tallow, 59.9
% Granulated sugar, 4.0% mineral blend, 0.85% vitamin blend, 4.0% filter paper powder, 1.0% cholesterol and 0.25%
They were raised on a high cholesterol diet consisting of bile acids. Two of the three groups receiving the high cholesterol diet were fed with a diet to which the mixture of sesamin and episesamin used in Example 2 was further added at 1.0% and 2.0%. 2
After breeding for one week, partial blood collection in the first week and total blood collection in the second week were performed after a 17-hour fast, and GOT and GPT activities in the serum were measured. Table 3 shows the results. The elevation of GOT by a high cholesterol diet could be reduced by giving a sesamin and episesamin mixture.

Example 4 A 9-week-old male CDF1 mouse (Clear Japan) was bred for 1 week, and the mice were divided into 3 groups, each group consisting of 7 mice. Except for the control group, the remaining 2 groups were exposed to air. Housed in a chamber prepared to contain 12 ppm ethanol. After bred for one week, blood is collected after a 16-hour fast, and total cholesterol in serum (T-CHO)
The amount, triglyceride (TG), GOT, GPT, and total bilirubin activities were measured. Table 4 shows the results.

Triglyceride amount in serum by alcohol administration,
Elevated total bilirubin, GOT and GPT could be significantly reduced by giving sesamin. Observation of general behavior also showed a marked difference, such as 5 out of 7 rats on the normal diet, which underwent alcohol administration, whereas those on the diet containing sesamin showed almost normal behavior. Was done.

Example 5 An 8-week-old male CDF1 mouse (Clear Japan) was bred for 1 week, and the mice were divided into three groups, each group consisting of seven mice. Except for the control group, the remaining two groups contained tetrachloride. One mg / kg of carbon was intraperitoneally administered, and one group was further gavaged with 100 mg / kg of sesamin at the same time. After administration, the animals were fasted for 16 hours, blood was collected, and total cholesterol (T-CHO) in serum, Triglyceride (TG)
The amount, as well as GOT, GPT, and total bilirubin activity were measured. Table 5 shows the results.

Example 6 Male Wistar rats were divided into 2 groups (control group, sesamin group) as 6 rats per group. The control group was bred on a basal diet, and the sesamin group was bred on a basal diet supplemented with 0.5% sesamin for 22 days. After an overnight fast, 1 g / kg of ethanol (20%) was given by oral gavage. One or three hours later, blood was collected from the tail vein and the blood alcohol concentration was measured.
One hour after ethanol administration, control group, 0.5 mg / ml
In contrast, the sesamin group was 0.27 mg / ml, and after 3 hours, the sesamin group was 0.02 mg / ml compared to the control group 0.12 mg / ml, and the sesamin group showed faster disappearance of ethanol from blood in the control group. Was seen.

Formulation Example 1 0.5 g of the compound of the present invention was mixed with 20.5 g of silicic anhydride, and 79 g of corn starch was added thereto, followed by further mixing. To this compound was added 100 ml of a 10% ethanol solution of hydroxypropylcellulose, and the mixture was extruded, extruded and dried as usual to obtain granules.

Formulation Example 2 7 g of the present compound was mixed with 20 g of silicic anhydride, and 10 g of microcrystalline cellulose, 3.0 g of magnesium stearate,
Lactose (60 g) was added and mixed, and the mixture was tableted with a single-shot tableting machine to produce tablets having a diameter of 7 mm and a weight of 100 mg.

Formulation Example 3 2.5 g of the compound of the present invention was added to a nonionic surfactant, TO-10.
M (Nikko Chemicals) dissolved in 200g at 122 ° C.
4.7975 of sterile physiological saline heated to 60 ° C. was added and stirred well, and the mixture was aseptically distributed in vials and sealed to produce an injection.

Continued on the front page (72) Inventor Yoshinori Kitagawa 1-1-1 Wakayamadai, Shimamoto-cho, Mishima-gun, Osaka Prefecture Inside the Basic Research Laboratory of Suntory Limited (72) Inventor Michihiro Sugano 5-38- Nashima, Higashi-ku, Fukuoka City, Fukuoka Prefecture 23 (72) Inventor Hideaki Yamada 19-1 Matsugasaki Kinohonmachi, Sakyo-ku, Kyoto-shi, Kyoto (72) Inventor Masaru Shimizu 14 Nishinokyo-Hakuraku-cho, Nakagyo-ku, Kyoto-shi, Kyoto 14 (56) References JP-A-63-207389 (JP, A) JP-A-63-165396 (JP, A) JP-A-61-212518 (JP, A) (58) Fields investigated (Int. Cl. ⁷ , DB name) A61K 31/34 A61K 35/78 C07D 493 / 04 CA (STN) MEDLINE (STN) REGISTRY (STN)

Claims (2)

(57) [Claims] 1. The following general formula (I): (Wherein R ¹ , R ² , R ³ , R ⁴ , R ⁵ , and R ⁶ are each independently a hydrogen atom, an alkyl group having 1 to 3 carbon atoms, or R ¹ and R ² , and / or R ⁴ And R ⁵ together represent a methylene group or an ethylene group, and n, m, l represent 0 or 1). Dioxabicyclo [3.3.0] octane derivative represented by the following formula: Agent. 2. The dioxabicyclo [3.3.0] octane derivative is sesamin, sesaminol, episesamin, episesaminol, sesamolin, 2- (3,4-methylenedioxyphenyl) -6- (3-methoxy- 4-hydroxyphenyl) -3,7-dioxabicyclo [3.3.0] octane, 2,6-bis- (3-methoxy-4-hydroxyphenyl) -3,7-dioxabicyclo [3.3.0] octane Or 2- (3,4-methylenedioxyphenyl) -6- (3
The liver function-improving agent according to claim 1, which is -methoxy-4-hydroxyphenoxy) -3,7-dioxabicyclo [3.3.0] octane.