JPH0477431A - Interferon-inducing agent - Google Patents

Abstract

PURPOSE:To obtain an interferon-inducing agent exhibiting excellent interferon- inducing action and low toxicity, and useful as a preventive and treating agent for viral diseases and cancer by using an extract of KAMI-SHOYO-SAN as an active component. CONSTITUTION:The objective interferon-inducing agent contains, as an active component, an extract (preferably concentrated extract or dried extract powder) of KAMI-SHOYO-SAN (a kind of formulated Chinese drug) effective for various diseases accompanying neural symptoms such as indefinite complaint and menopausal disturbance. It has excellent interferon (IFN) inducing action compared with KOSAIKO-TO (a kind of formulated Chinese drug) known as a Chinese drug having IFN-inducing activity and, accordingly, the agent can be effectively and safely used as an antiviral agent and a carcinostatic agent. The daily dose of the agent is usually 0.3-10g of the dried extract powder for adult in single dose or 2-3 divided doses by oral administration.

Description

[Detailed description of the invention] [Industrial application field] The present invention relates to interferon inducers.

More specifically, the present invention relates to an interferon inducer containing an extract of Kamishoyosan as an active ingredient.

[Prior Art] Interferon (hereinafter referred to as IFN) is a cytokine with the most advanced clinical application, and is used as an antiviral agent and an anticancer agent. It is also known that various substances induce (endogenous) IFN, and such substances (IFN-inducing agents) can also be used as antiviral agents and anticancer agents like IFN.

Kampo medicines are widely used for chronic diseases as they have relatively few side effects, but among these Kampo medicines, it has been reported that Shosaikoto has an IFN-inducing effect [Medicine and Pharmacy] , 10(2):687-693.19
83, Proceedings of the 109th Annual Meeting of the Pharmaceutical Society of Japan V, p. 85,
Nagoya, 1989].

[Problems to be Solved by the Invention] The present inventors conducted various studies in order to find a new Chinese herbal medicine that exhibits a better IFN-inducing effect than Shosaikoto.

An object of the present invention is to provide a new Chinese herbal medicine that exhibits excellent IFN-inducing activity.

[Means for Solving the Problems] As a result of studying various Chinese herbal medicines, the present inventors have developed an extract of Kamishoyosan (Kamishoyosan), which has been used for various diseases accompanied by neurological symptoms such as indeterminate complaints and menopausal disorders. The present invention was completed by discovering that a drug (hereinafter referred to as the drug of the present invention) satisfies the above objectives.

The composition (mass ratio) of Kami Shoyosan in the present invention is (2
, 0-4.0), Xun Yao (2,0-4.(1), White Tr,
(2, o ~ 4.0), Kyo Ling (2,0 ~ 4.0), Chai Hu (
2.0-4.0), Botanpi (1.0-3.0), Yamateko (1.0-3.0), Licorice (0.5-3.0), Seikyo (
0.5 to 3.0), Hogane (1.0 to 2.0), and particularly preferred are Toki (3,0), Enyaku (3, no, Shiro* (
3,0), Hagirei (3,0), Saiko (3,0), Botanpi (
2,0), Yamateko (2,0), Licorice (1,5-2.0)
, major (0,5-2.0), and minor (1,0).

In the present invention, it is preferable to extract Kamishoyosan with a solvent and use it as a concentrated extract or dry extract powder.

The drug of the present invention can be produced as follows.

First, add an extraction solvent of 5 to 25 times, preferably 8 to 20 times, in weight ratio to Kamishoyosan.
The extract is heated at 00°C for 30 minutes to 2 hours to obtain an extract. The extraction solvent is water, a water-soluble organic solvent, or a mixed solvent thereof, and ethanol is preferable as the water-soluble organic solvent.

Next, the extract is filtered or centrifuged, and then the drug of the present invention is purified by conventional concentration means such as vacuum concentration, and if necessary further conventional drying means such as vacuum drying, spray drying, or freeze drying. can get.

The drug of the present invention obtained as described above can be used as is, but if necessary, it may be added with conventional pharmaceutical additives such as excipients and disintegrants, such as lactose, starch, crystalline cellulose, carboxymethyl cellulose, calcium, It can also be formulated into capsules, granules, fine granules, tablets, or powders by a conventional method by adding silicic anhydride, magnesium stearate, and the like.

As shown in the test examples below, the drug of the present invention exhibits a strong IFN-inducing effect and has low toxicity, and can be used as a prophylactic and therapeutic drug for viral diseases and cancer.

It is used for viral diseases such as chronic hepatitis B, and cancers such as renal cancer, multiple myeloma, malignant melanoma, malignant lymphoma, and adult T-cell leukemia.

The dosage of the drug of the present invention varies depending on the patient's condition, age, weight, etc., but it is usually 0.3 to 10 g as a dry extract powder per day for adults at once or 2 to 3 times per day.
Administer orally in divided doses.

[Actions and Effects of the Invention] The drug of the present invention has a superior IFN-inducing action to Shosaikoto (see Test Examples 1 and 2).

The fact that the above-mentioned effect is not a direct effect of the drug but is based on the (endogenous) inducing action of IFN is that when measuring IFN activity in Test Examples 1 and 2, rabbit anti-mouse IFN-α/β was added to the specimen sample. This was confirmed by the fact that its activity was neutralized when it was coexisting with antibodies.

Furthermore, the toxicity of the drug of the present invention is low (see Test Example 3).

Therefore, the drug of the present invention can be used effectively and safely as an excellent IFN inducer for the prevention and treatment of viral diseases and cancer.

The pharmacological tests related to the drug of the present invention will be described below.

Test Example I IFN by IFN-induced action (in vitro): (1) Sample powder of dried shoyosan extract of Sample Example 1 (drug of the present invention) and dried powder of Shosaikoto extract of Comparative Example 1 (control drug).

(2) Test method Spleen cells were collected from C3H/He female mice (8 weeks old), and 10 (W/V)% fetal bovine serum (NU-3ERUM,
Col 1a-volative Re5search
Inc.) and RPMI-1640 medium (Nippon Pharmaceutical) containing 1 kg/ml gentamicin.
'cells/ml spleen cell suspension was prepared. To 0.5 ml of this cell suspension, add 0.5 ml of the above medium containing 200 g/ml of the specimen, and add it to a 24-well culture plate (Fa
lcon 3047, Becton Dickinso
After culturing at 37°C for 24 hours at 37°C, the culture supernatant was centrifuged (1000 rpm, 10 minutes), and the supernatant was collected and used for IFN activity measurement.

Measurement of IFN activity due to the IFN-inducing effect of the sample was performed using a 50% cytopathic effect inhibition method (Interferon Science, p. 13, edited by Shigeyasu Kobayashi, Kodansha) using mouse L929 cells.

First, prepare a 96-well culture plate (Falcon 307
2, Be-cton Dickinson Inc.) with 5 L929 cells (4 x 10'/well) (W/V
)% NN0-3ERO-containing MEM medium (Nippon Pharmaceutical Hoogt) to form a monolayer overnight, and the above (
The sample solution for IFN activity measurement obtained in step 1) was diluted with 0.5 (
W/V)% NU-8ERUM in MEM medium containing 2-fold serial dilutions and cultured at 37°C overnight.
is virus, Livestock Hygiene Laboratory) was added to L929 cells as a challenge virus. After overnight incubation at 37°C,
The IFN activity caused by the IFN-inducing effect of the sample was measured using the inhibition rate of the cytopathic effect of L929 cells as an index.

In the above test, the unit of IFN activity is the 5% of the cytopathic effect when no sample is added (IFN-untreated cells).
It was expressed as the reciprocal of the dilution indicating 0%.

In addition, IFN activity was measured using standard IFN (Lee Bio Mo
1ecularResearch Laboratory
ies Inc.) were tested at the same time, and the IFN international unit [1 international unit (IU)/m
l].

(3) Test results The test results are shown in Table 1 below together with the results of Test Example 2 (in vivo).

Test Example 2 Measurement of IFN activity by IFN-induced effect (in vivo)
: (1) Same as sample test example 1.

(2) Test method C3H/He female mice (8 weeks old, 3 mice in - group) were intraperitoneally injected with 0.2% of the sample (100 μ/kg) in a phosphate buffered saline solution.
ml was administered, and 14 hours after administration, whole blood was collected from each mouse individually, and the serum was used for measuring IFN activity.

Measurement of IFN activity based on the IFN-inducing effect of the sample was carried out in the same manner as in Test Example 1.

(3) Test results The test results are shown in Table 1.　　　　　,.

Table 1 Test Example 3 Acute toxicity test: (1) Specimen and test method Add 0.5 (W/V) of the seasoned Shoyo San dried extract powder of Example 1.
% carboxymethyl cellulose suspended in sodium aqueous solution (concentration 1000 μ/ml) 10 g to ddY male mice (6 weeks old, weight 26-29 g, 10 mice per group)
The number of deaths was observed for up to 2 weeks after administration.

(2) Test results: No deaths were observed within 2 weeks after administration; LD of Kami Shoyo San dried extract powder. The value was >Log/kg.

Therefore, the drugs of the present invention have extremely low toxicity.

[Example code] Next, the present invention will be explained with reference to Examples and Comparative Examples.

Example 1 Dried extract powder of seasoned shoyosan: 3.0 kg each of Dangki, Xun Yaku, Shiraki, Hagi Ling, and Chai Hu, 2.0 kg each of Mudanpi and Sanzi, 1.5 kg of licorice, main Water 2201 was added to a mixed herbal medicine consisting of 0.5 kg and 1.0 kg, and the mixture was heated and extracted at about 100°C for 1 hour.

The extract was filtered, and the filtrate was concentrated under reduced pressure to about 20 ml, and then spray-dried to obtain 4.1 kg of seasoned shoyo-san dried extract powder.

Example 2 Flavored Shoyo powder and two sets of granules: (Formulation) Composition Blend amount Main drug (dry extract powder of Example 1) 4.10 parts by weight Lactose 0.14 parts by weight Corn starch 1.30 parts by weight Anhydrous silicon Acid: 0.37 parts by weight Magnesium stearate: 0.09 parts by weight (Operation) The above components were thoroughly mixed, and this mixture was formed into a plate using a compression molding machine, then ground into granules using an oscillator, and sieved for size. Kami Shoyosan extract fine granules containing 683 ml of crude drug per gram were obtained.

Example 3 Kamimi Shoyosan extract tablets: (Formulation) Composition Crude drug (dry extract powder of Example 1) Lactose Corn starch Synthesis Aluminum silicate Carboxymethylcellulose Calcium Magnesium stearate (Procedure) The above components were thoroughly mixed, and this mixture was is compressed into tablets containing 3.00 parts by weight, 1.00 parts by weight, 0.50 parts by weight, 0.20 parts by weight, 0.25 parts by weight, 0.05 parts by weight, and 300 mg, and each tablet contains 180 kg of the active ingredient. A tablet containing Kamishoyosan extract was obtained.

Example 4 Kami Shoyosan extract capsules: (Formulation) Composition Blend amount Main ingredient (dry extract powder of Example 1) 3.34 parts by weight Synthetic aluminum silicate 0.18 parts by weight Magnesium stearate 0.08 parts by weight ( Procedure) The above-mentioned components were thoroughly mixed and 360 ml of this mixture was filled into capsules to obtain Kamishoyosan extract capsules containing 334 mg of the crude drug in each capsule.

Comparative Example 1 Shosaikoto dry extract powder: Saiko 7. Okg, half summer 5.0kg, 3.0kg each of Huanglium, Taijutsu, and Carrot, licorice 2. Okg and major 1.0
Add water 2401 to a mixed herbal medicine consisting of 1 kg and boil at about 100°.
The mixture was heated and extracted at C for 1 hour. Filter the extract and remove the filtrate from approx.
After concentrating under reduced pressure to a concentration of 0.01 kg, the mixture was spray-dried to obtain 5.4 kg of Shosaikoto dry extract powder.

Claims (1)

[Claims] An interferon inducer whose active ingredient is the extract of Kamishoyosan.