JPH0159248B2 - - Google Patents

Description

[Detailed description of the invention]

The present invention relates to antihepatitis agents. The present inventor has spent many years searching for substances that have antihepatitis effects, especially hepatitis preventive effects, and has conducted screening tests on numerous substances.As a result, Tendai Toriyaku is one of the herbal medicines that have been used medicinally since ancient times. The present invention has been completed by discovering that Tendaiyaku, Gajiyutsu, and Byakujiyutsu have a novel medicinal effect: antihepatitis. Tendai toriyaku [scientific name: Linderae Radix, family: Lauraceae] is native to central and southern China, and the long, spindle-shaped part of the root is used for medicinal purposes. It is used to treat symptoms of functional decline. Also, 〓 [scientific name:
Zedoariae Rhizoma,
The ginger family contains essential oils including cineole, sesquiterpene alcohol, camphene, etc., as well as other oils such as fatty oils, starchy substances, mucilage substances, and rubber substances, and is used as an aromatic stomachic medicine as a raw material for home remedies, and is sometimes used as a stimulant for young children. It is used to treat symptoms of high fever. In addition, the roots of white perch (scientific name: Atractylodis Rhizoma, Asteraceae) are harvested and dried, and are used as a stomachic medicine and in the production of tososan. . Therefore, it has not been previously known that Tendai Wuyaku, Porphyra and Baekju have an antihepatitis effect, and this was discovered for the first time by the present inventors. As a result of extracting and purifying the active ingredients from these herbal medicines and analyzing them, we found that the substances that have antihepatitis effects are serinane-type sesquiterpenes such as lindesthrene, lindelene, siedeurone, and atractilone, and germacrane-type sesquiterpenes such as lindelane and linderlactone. It was confirmed that it was a terpene. The anti-hepatitis effect is not limited to the serinane-type sesquiterpenes and germacrane-type sesquiterpenes themselves, but also the organic solvent extracted extracts of Tendai toriyaku, 莪〓, and Hakushu, or the organic solvent extracted extracts of these herbal medicines directly. The essential oil obtained by steam distilling the extract also has anti-hepatitis effects. The serinane-type sesquiterpenes and/or the serinane-type sesquiterpenes of the present invention
The pharmacological effects and acute toxicity of extracts extracted with organic solvents and essential oils obtained by steam distillation, which contain germacrane sesquiterpenes as active ingredients, will be explained. As for the pharmacological effects, the extracts extracted with organic solvents or the essential oils obtained by steam distillation produced in Production Examples 1 to 6 shown in Table 1 below were used as test drugs, and (i) the effect of preventing the onset of carbon tetrachloride liver damage; ii) The effect of preventing the onset of galactosamine liver damage and (iii) The effect of preventing the onset of ethionine liver damage were tested.

[Table] In addition, in both studies, α-mercaptopropionylglycine (trade name: Thiola, hereinafter referred to as α-MPG) and protoporphyrin sodium (hereinafter referred to as α-MPG), known substances with similar medicinal efficacy, were used.
NAPP. ) was used as a control drug. (i) Preventive effect on the onset of carbon tetrachloride liver damage Each test drug was mixed and suspended with gum arabic powder and administered orally twice a day for 3 days to a group of 10 DD male mice weighing 18 to 20 g. At the same time as the fifth administration, 2 ml of carbon tetrachloride 1.2% olive oil solution was injected subcutaneously per 1 kg of body weight. One hour after the sixth administration, blood was collected from the abdominal aorta, and GOT and GPT in the serum were collected. The value was measured. Table 2 shows the results of a comparison between the carbon tetrachloride untreated group and the carbon tetrachloride treated group as a control group.

[Table] * Mean value ± standard error Serinane-type sesquiterpenes of the present invention and/or
Extracts and essential oils containing germacrane-type sesquiterpenes as active ingredients are effective against GOT and GPT, which are observed in carbon tetrachloride liver injury in mice.
It suppressed the rise in levels, and was more effective at a smaller amount than α-MPG and NAPP, which have the same known efficacy, and the essential oil content was particularly effective. (ii) Preventive effect of galactosamine on liver damage 10 male Wistar rats weighing 180-200g
Each test drug was mixed and suspended in gum arabic powder and administered orally to one group of animals. One hour later, 300 mg of galactosamine hydrochloride per 1 kg of body weight was administered intraperitoneally. After 20 hours, blood was collected from the abdominal aorta. GOT and GPT values in serum were measured. Table 3 shows the results of comparing the non-galactosamine treated group and the galactosamine treated group as a control group. Serinane-type sesquiterpenes and/or serinane-type sesquiterpenes of the present invention
Extracts and essential oils containing germacrane-type sesquiterpenes as active ingredients were found to have the same effect on preventing liver damage as α-MPG and NAPP used as control drugs, and in particular, the essential oil content was 1/1/2 that of the control drugs. Similar effects were obtained at a dose of 3.

[Table] * Mean value ± standard error
(iii) Preventive effect on the onset of ethionine liver damage 10 male Wistar rats weighing 180-200g
Each test drug was mixed and suspended with gum arabic powder in groups of animals, and administered orally. One hour later, ethionine was administered intraperitoneally at 1000 mg per 1 kg of body weight. After 20 hours, blood was collected from the abdominal aorta and serum of
GOT and GPT values were measured. Table 4 shows the results of comparing the ethionine-untreated group and the ethionine-treated group as a control group.

[Table] * Mean value ± standard error Serinane-type sesquiterpenes of the present invention and/or
Or extracts and essential oils containing germacran type sesquiterpenes as active ingredients are α-
Serum GOT, equivalent to MPG and NAPP,
It has the effect of suppressing the rise in GPT levels, and in particular, the essential oil content showed an effect equal to or greater than that of the control drug at a small dose. (iv) Acute toxicity Each test drug and control drug were orally administered to a group of 15 DD male mice weighing 18 to 20 g, and the LD ₅₀ was determined from the number of deaths over 3 days. Water during the observation period;
Both feeds were provided ad libitum. The results are shown in Table 5. Even at a dose of 1500 mg/kg body weight, there was no fatality in any case, and therefore, the extracts and essential oils containing serinane-type sesquiterpenes and/or germacrane-type sesquiterpenes as active ingredients of the present invention are not acutely toxic. It is clear that it is weak.

[Table] As is clear from the above test examples, organic solvent extracted extracts and essential oils containing serinane-type sesquiterpenes and/or germacrane-type sesquiterpenes as active ingredients exhibit antihepatitis effects when administered orally. It can be demonstrated. When used for the treatment of adults, considering the results of the above studies, it is estimated that 50 to 150 mg at a time, 2 to 3 times a day, is effective; however, in the case of organic solvent extracts, It is considered preferable to take 100 to 150 mg at a time, 2 to 3 times a day, and in the case of essential oils, 50 to 100 mg at a time, 2 to 3 times a day. For administration, it is desirable that the drug be provided in a form suitable for absorption from the gastrointestinal tract. Next, a method for producing an antihepatitis agent containing serinane-type sesquiterpenes and/or germacrane-type sesquiterpenes from crude drugs as an active ingredient will be described. Tendai toriyaku, 莪〓, or Hakushu is immersed in a non-polar organic solvent or a polar organic solvent to extract the active ingredients, and then filtered and concentrated to produce serinane-type sesquiterpenes and/or germacran-type sesquiterpenes. An extracted extract containing 5% or more is obtained. As the nonpolar organic solvent, n-hexane, ether, petroleum ether, ethyl acetate, etc. can be used, and as the polar organic solvent, methanol, ethanol, etc. can be used. Extraction from crude drugs is performed by cold immersion or wet immersion. Concentration is performed by methods such as vacuum concentration and ultrafiltration, and if necessary, the organic solvent is distilled off to dryness or refrigeration is carried out. When producing an antihepatitis agent containing serinane-type sesquiterpenes and/or germacrane-type sesquiterpenes as active ingredients by steam distillation, the above-mentioned herbal medicines, Tendai toriyaku, 莪〓, or Hakushu are directly steam distilled. The distillate is subjected to operations such as shaking and separation using a separatory funnel to obtain the essential oil component, which is the active ingredient. or,
In addition to directly using the herbal medicine, it is also possible to obtain the essential oil, which is the active ingredient, by steam distillation using the organic solvent extract of Tendai toriyaku, 莪〓, or Hakushu as described above. The essential oil contains 10% or more of serinane-type sesquiterpenes and/or germacrane-type sesquiterpenes, and according to the results of the pharmacological tests mentioned above, it has a stronger medicinal effect than the organic solvent extracted extract. The physical properties of the above extracted extract and essential oil are as follows. It is a pale yellow to brown oily or viscous substance with a slightly bitter taste and aroma, and is insoluble in water. These were prepared using silica gel G as an adsorbent.
The figure shows the pattern of thin layer chromatography developed using a developing solvent of -hexane:ethyl acetate = 10:1 and colored with Ehrlich's reagent. Furthermore, serinane-type sesquiterpenes and/or germacrane-type sesquiterpenes that were positive for Ehrlich reagent were analyzed by gas chromatography (equipment: Hitachi model 163,
Detection tube: FID, Separation tube: 1.5% OV1, Separation temperature:
140℃, carrier gas: nitrogen gas 25ml/min,
Integrator: Takeda Riken TR-2217). From the above results, the serinane-type sesquiterpenes of the present invention are lindesrene, lindelene,
It was confirmed that these were siedeurone and atractilone, and the germacrane-type sesquiterpenes were linderane and linderlactone. As mentioned above, antihepatitis drugs containing serinane-type sesquiterpenes and/or germacrane-type sesquiterpenes contained in herbal medicines such as Tendai Toryaku, 莪〓, or Hakushuo as active ingredients have a dosage form as shown in the formulation example. When administered orally, it exhibits excellent anti-hepatitis effects and is extremely useful as a new anti-hepatitis agent.In addition, the method of extraction from crude drugs is relatively simple, and raw crude drugs can be easily obtained. Since it is available and relatively inexpensive, it has high industrial applicability, with low manufacturing costs. Next, the present invention will be specifically explained by showing production examples and composition examples. Production Example 1 Petroleum ether (5) was added to 1 kg of coarsely chopped Tendai toriyaku, digested three times, and the extract was concentrated and dried to obtain 200 g of dry extract. This dried extract was analyzed by thin layer chromatography and gas chromatography and was found to contain 8.9% in total of serinane-type sesquiterpenes and germacrane-type sesquiterpenes, which are positive with Ehrlich's reagent. Production Example 2 N-hexane (5) was added to 1 kg of coarsely chopped cucumbers, digested three times, and the extract was concentrated and dried to obtain 21.8 g of dry extract. When this dry extract was quantitatively determined in the same manner as in Production Example 1, it contained 15.8% of the total amount of serinane type and germacran type sesquiterpenes. Production Example 3 1 kg of coarsely cut white persimmon was added with 5 n-hexane and digested three times, and the extract was concentrated and dried to obtain 18.5 g of dry extract. This dried extract contained 20% of atractylone, a serinane-type sesquiterpene, when determined in the same manner as in Production Example 1. Production example 4 Add methanol (5) to 1 kg of coarsely chopped Tendai toriyaku, digest it three times, concentrate under reduced pressure while keeping the temperature below 50°C, distill off the solvent until it becomes candy-like, and then add to this candy-like concentrate. Water from step 2 was added and steam distilled to obtain 11 ml of essential oil. When the essential oil was dried over anhydrous magnesium sulfate and quantified in the same manner as in Production Example 1, it contained 42.5% of the total amount of serinane type and germacran type sesquiterpenes. Production example 5 Add 5 methanol to 1 kg of coarsely chopped cucumbers,
Digested three times and concentrated under reduced pressure while keeping the temperature below 60°C to obtain a candy-like concentrate. Dissolve this in a small amount of methanol, add n-hexane from step 5 and repeat the shaking separation three times to obtain the n-hexane extract. Concentrate and dry,
17 g of n-hexane extract was obtained. When this extract was quantitatively determined in the same manner as in Production Example 1, it contained 35.5% of the total amount of serinane type and germacran type sesquiterpenes. Production example 6 Add 20 methanol to 5 kg of coarsely chopped white persimmon,
Digested three times and concentrated under reduced pressure while keeping the temperature below 60℃ to obtain a candy-like concentrate, which is then steam distilled to extract the essential oil.
Obtained 80g. When this essential oil was quantitatively determined in the same manner as in Production Example 1, it was found to contain 45% atractylone, a serinane-type sesquiterpene. In addition, throughout the above production examples, the temperature should be 100°C or less, preferably 60°C or less. Formulation example 1 Liquid preparation Essential oil obtained in Production Example 4 100mg of olive oil 200mg Total 300mg (for 1 vial) Completely dissolve the essential oil in olive oil, fill in a vial, and seal. Production Example 2 Essential Oil Potato Starch Magnesium Stearate Obtained in Tablet Production Example 4 10.0g 19.5g 0.5g Total 30g (100 tablets) The essential oil and potato starch were mixed, water was added, and kneaded. It is made into granules through a granulator with a 1 mm x 1 mm mesh screen, and after drying, No. 16
Sort the grains using a Metsushiyu (BS) sieve. Next, the granules were mixed with magnesium stearate and formed into tablets of 300 mg each using a tablet press. Further, the tablets were coated with a conventional easily soluble film coating, if necessary.

[Brief explanation of drawings]

The figure shows thin layer chromatography patterns of extracts or essential oils obtained in Production Examples 1 to 6 of the present invention.

Claims (1)

[Scope of Claims] 1. An antihepatitis agent containing as an active ingredient serinane-type sesquiterpenes, germacrane-type sesquiterpenes, or a mixture of serinane-type sesquiterpenes and germacrane-type sequiterpenes obtained from crude drugs. 2. The anti-hepatitis agent according to claim 1, wherein the crude drug is Tendai toriyaku. 3. The anti-hepatitis agent according to claim 1, wherein the herbal medicine is Gajiyutsu. 4. The antihepatitis agent according to claim 1, wherein the crude drug is Byakujiyutsu.