JPH0477428A - Agent for suppressing proliferation of aids virus - Google Patents

Abstract

PURPOSE:To provide the subject agent containing hetero-annular diene glycyrrhizin as an active component, exhibiting remarkable proliferation suppressing action on AIDS virus and having little side effect. CONSTITUTION:The objective agent contains olean-11,13(18)-dien-3beta-ol-30oic acid 3-O-beta-D-glucuronopyranosyl(1$22)beta-D-glucurono-pyranoside (hetero-annular diene glycyrrizin) expressed by formula or its pharmacologically permissible salt as an active component. The compound is a modification of glycyrrhizin existing in licorice and known to have antiinflammatory, anti-allergic or anti- ulcer action. Various effects such as prevention of cricis of AIDS, mitigation of various symptoms and elimination of symptoms can be expected by administering the present agent for suppressing proliferation of AIDS virus by oral or parenteral administration at a daily dose of 200-800-mg for adult.

Description

DETAILED DESCRIPTION OF THE INVENTION (Industrial Field of Application) The present invention relates to an AIDS virus growth inhibitor containing a modified compound of glycyrrhizin, which is an active ingredient of licorice, as an active ingredient.

(Prior art and its problems) Acquired immunodeficiency syndrome (AIDS) is caused by a virus, and this virus
Immunodeficiency virus (HI
V). This virus belongs to the human retrovirus group, and this virus destroys the helper T cells of lymphocytes, resulting in cellular immunodeficiency, resulting in opportunistic infections caused by protozoa, fungi, viruses, bacteria, etc., and Kabosi's sarcoma.

The present applicant provided an AIDS virus growth inhibitor containing glycyrrhizin contained in licorice as an active ingredient in Japanese Patent Application Laid-Open No. 63-33332. Subsequently, in the treatment of AIDS with glycyrrhizin, doses ranging from several times to several tens of times as much as when glycyrrhizin is used to treat hepatitis etc. may be administered to AIDS patients and carriers for a long period of time.
There was great concern that glycyrrhizin might cause pseudoaldosteronism, a side effect. What is pseudoaldosteronism?
Guttyrrhizin is hydrolyzed in the body to produce the aglycone glycyrrhetinic acid, which inhibits the metabolism of steroid-metabolizing enzymes in the liver, resulting in abnormal electrolyte metabolism, hypertension, or hypokalemia.

Therefore, the inventors of the present application searched for various substances that have the effect of suppressing the growth of the AIDS virus without the side effect of developing pseudohyperaldosteronism, such as glycyrrhizin, and found that they have the same effect of suppressing the growth of the AIDS virus as glycyrrhizin, and We have discovered a glycyrrhizin-modified compound that has almost no side effects.

This is heteroannuular dienglycyrrhizin, which was disclosed by the applicant in Japanese Patent Publication No. 63-2959, and is known to have anti-inflammatory, anti-allergic, or anti-ulcer effects, but it is known to have anti-inflammatory, anti-allergic, and anti-ulcer effects. No inhibitory effect was known. However, the above-mentioned compound was confirmed by the inventors to have a remarkable effect of inhibiting the proliferation of the AIDS virus, as described below.

(Structure of the Invention) The AIDS virus proliferation inhibitor according to the present invention comprises 11.13(18)-dienglycyrrhizin (
The active ingredient is heteroanular dienglycyrrhizin.

This compound can be obtained by the method described in Japanese Patent Publication No. 63-2959. That is, it is obtained by adjusting an aqueous alkaline salt solution of 11β-hydroglycyrrhizin to pH 7 to 8 with a mineral acid, heating it, and then adjusting the pH to 2 to 3. Alternatively, it can be dissolved in methanol and reacted with a corresponding amount of an alkali agent, and then acetone is added to the reaction solution to obtain an alkali salt.

Details will be described later as a reference example.

The above-mentioned heteroannular dienglyderritin and its pharmaceutically acceptable alkali addition salt have a remarkable effect of inhibiting the proliferation of the AIDS virus, as shown in the experimental examples below, and can be used as an AIDS virus proliferation inhibitor as is. or a pharmaceutically acceptable salt thereof, that is, a glycyrrhizin salt such as glycyrrhizin ammonium salt, glycyrrhizin alkali metal salt, or glycyrrhizin choline salt obtained by reacting glycyrrhizin with an inorganic or organic base as an active ingredient. , can be administered to AIDS patients.

Either oral or parenteral administration can be selected, and the dosage depends on the patient's symptoms, i.e. AC, ARC.
, depending on the stage of AIDS, adults - 20 per day
By using it in the range of 0 to 800 mg, effects such as prevention of the onset of AIDS, alleviation of various symptoms, and disappearance of symptoms can be expected.

The drug layer contains one or more pharmaceutically acceptable harmless excipients, such as sodium bicarbonate, sodium chloride, lactose, potato starch, sodium alginate, magnesium silicate, calcium carbonate, hydroxypropyl cellulose, etc. It can be made into powders, granules, sugar-coated tablets, capsules, etc. by conventional methods.

The solvent for injections is, for example, physiological saline or 5%
Glucose solution is suitable.

Examples of formulations are listed below.

[Margins below] [Formulation Examples] Heteroannular dienglycyrrhizin precipitated calcium carbonate crystalline cellulose talc 00 g 00g 00g 0g The above composition was prepared by a conventional method until the weight per tablet was 250.
It was made into a mg tablet.

Examples of experiments regarding the inhibitory effect of heteroannular dienglycyrrhizin on the proliferation of AIDS virus are listed below. These experiments revealed that this compound strongly suppresses the proliferation of the AIDS virus (HIV) and has the effect of suppressing the degeneration of infected cells.

[Experimental example] (Materials) (1) Cells and virus-infected cells used were MT-4 cells. The virus is Mo1t
-4/HIV producing strain was used.

(2) Experiment for suppressing degeneration of infected cells This experiment was conducted using heteroannual dienglycyrrhizin.
The present invention relates to the inhibitory effect of HIV-infected cell degeneration on o1r-4No8 cells.

2 x 10' (7) MI-4 cells, 6 x 10'' PFU
(7) The cells were infected with HIV, added with various concentrations of heteroannuular dienglycyrrhizin solutions, and cultured for 3 days in RPM11640 culture medium containing antibiotics and 10% fetal bovine serum.

Thereafter, the above-mentioned culture solution was stirred and divided into halves, and aqueous heteroannuular dienglycyrrhizin solutions of various concentrations mixed in the culture solution were added to one half, and cultured for an additional 2 days.

As a control, a similar experiment was performed on uninfected cells.

After culturing, the number of living cells was measured by the trypan blue exclusion method. The results are shown in FIG. In the figure, each mouth is HI.
V uninfected cells, % indicates HIV infected cells, cells were infected with HI
When infected with V, the number of viable cells decreases to about 29% of the initial number, but when cultured with the addition of heteroannuular dienglycyrrhizin, the number of viable cells increases. In this case, the concentration of heteroannulus glidellidene is 0.16 m
Efficacy can be confirmed at M or higher, and approximately 80% or more of the HIV-infected cells are viable. In this experiment, 1
．． It was effective at concentrations up to 24 mM, and significant cell growth inhibition was observed at higher concentrations.

From the above results, it can be seen that heteroannuular dienglycyrrhizin effectively suppresses the proliferation of HIV.

(3) Detection of virus antigen The presence of virus antigen in infected cells was confirmed by indirect fluorescent antibody method using anti-HIV antibody positive serum.

When heteroannual dienglycyrrhizin was not added, viral antigens could be detected in about 70% of cells due to HIV infection, but by adding heteroannuular dienglycyrrhizin, viral antigens gradually decreased.

As shown in FIG. 2, when the concentration of heteroannular dienglycyrrhizin is 0.16 mM, the viral antigen is about 30% or less, and as the concentration increases, the antigen gradually decreases.

(4) Acute toxicity test The test was carried out by intravenous injection into 8-week-old male ICR mice (body weight 36-38 g) according to Van der Waerden's method. The results are as follows.

L D a. :333-358 mg/kg [Reference Example] Production of heteroannuular dienglycyrrhizin (1) Dissolve sodium borohydride in an aqueous tetrahydrofuran solution containing a sodium hydroxide solution, and heat on a water bath at 80°C or higher to the boiling point of the solvent. Reflux.

A tetrahydrofuran solution of glycyrrhizin (manufactured by Gokensha Minophagen Seiyaku Honpo) was added dropwise to this, and a heating reaction was carried out for 24 hours. Add acetone and dilute hydrochloric acid to the reaction solution,
Decomposes excess alkali and reducing agent and adjusts the liquid to pH 9-1.
When adjusted to 0, glycyrrhizin is completely reduced and becomes the sodium salt of 11β-hydroglycyrrhizin.

Next, the above reaction solution was concentrated and a mixture of methanol and acetone was added to produce a white precipitate, which was collected by filtration and 75%
Recrystallization from ethanol yields 11β-hydroglycyrrhizine trisodium salt.

(2) Dissolve this in an appropriate amount of water, adjust its pH to 7 to 8 with dilute hydrochloric acid, and then heat it to quantitatively convert to the alkaline salt of heteroannuular dienglycyrrhizin. Dilute hydrochloric acid is added to the reaction solution to adjust the pH to 2 to 3, and the mixture is extracted with n-butanol and the solvent is distilled off to obtain a white crystalline powder of heteroannuular dienglycyrrhizin, which is the target product.

[Brief explanation of the drawing]

FIG. 1 is a diagram showing the results of an experiment on the degeneration suppressing effect of heteroannuular dienglycyrrhizin on Mr-4 cells in the experimental example of the present application, and FIG. 2 is a diagram showing the results of the virus antigen detection experiment. Figure 1Figure 2Con5entoratjon (9PIMlCon
ccntration (mM) Procedural Amendment Specification August 31, 1990

Claims (1)

[Claims] An AIDS virus growth inhibitor containing a glycyrrhizin-modified compound represented by the formula: ▲Mathical formula, chemical formula, table, etc.▼ and a pharmaceutically acceptable salt thereof as active ingredients.