KR0166570B1 - Pharmaceutical compositions for improvement of immune system and treatment of aids disease - Google Patents

Abstract

The present invention aids in the use of Bambusae Concretio Silicea, its water extract (Code No. 142), or its fractionated material (Code Nos. 142-2A, 142-3E and 142-3F). The present invention relates to a method for use in the treatment and immunopotentiation of (AIDS), a preparation containing the same as an active ingredient, and a method for producing the same, wherein the substances of the present invention have potent anti-HIV activity.

Description

AIDS treatment and immune function composition

FIG. 1 is a graph showing MTT foramazan production in MT-4 cell cultures treated with No. 142 extract.

Figure 2 is a graph showing MTT foramazan production in MT-4 cell cultures treated with No. 142-2A extract.

FIG. 3 is a graph showing MTT foramazan production in MT-4 cell cultures treated with No.142-3E extract.

Figure 4 is a graph showing MTT foramazan production in MT-4 cell cultures treated with No.142-3F extract.

FIG. 5 shows No. 1 for HIV-1 p24 antigen expression in MT-4 cells. Graph showing the inhibitory effect of 142-2A and AZT according to the culture day.

FIG. 6 shows No. 1 for HIV-1 proliferation inhibition in MT-4 cells by HIV-1 p24 antigen detection ELISA. Graph showing the effect of 142-3E.

FIG. 7 shows No. 1 for inhibition of HIV-1 proliferation in MT-4 cells by MTT assay and HIV-1 p24 antigen detection ELISA. Graph showing the effect of 142-2A.

FIG. 8 is a graph showing MTT formazan production in MT-4 cells after 1 hour pre-reaction with No. 142-2A and HIV-1. FIG. 9 shows MT-4 cells and HIV-1 in 1 hour in advance. After reaction, the graph shows MTT formazan production in MT-4 cells.

The present invention relates to a composition for treating AIDS and enhancing immune function.

In particular, the present invention relates to a composition for the treatment and immune strengthening of AIDS containing as a main component the Bambusae concretio Silicea (天竺 黄), its extract and fractions fractionated from the extract.

Since AIDS was first discovered in Los Angeles in 1982, AIDS has reported 1,025,073 AIDS patients (estimated 4.5 million) by the end of December 1944. Human immunodeficiency The virus (HIV) is estimated to reach 19 million people. In Korea, the number of AIDS patients has been steadily increasing since the discovery of American AIDS patients in 1985. As of the end of March 1995, the number of HIV infected patients was 432 (including 31 patients).

With the rapid increase in the number of HIV and AIDS patients, the US, France, Canada, Japan and other countries have spurred the development of therapeutic agents.As known anti-HIV substances, 2,3-azidothymidine has been approved by the US FDA. (AZT), dideoxyinosine (ddI), dideoxycytidine (ddC) and only a few. In addition to the occurrence of severe side effects, it is very difficult to expect the full therapeutic effect, the development of new drugs has been recognized as a very important task.

Therefore, the present inventors concentrated their research to find a substance showing fragranceal HIV activity in vitro by extracting components from natural products native to Korea and China as part of the development of AIDS therapeutics. Natural products have been shown to exhibit HIV activity.

According to the primary color natural drug dictionary published by Namsan Dang, Cheonggyelwang (Pixel Huang) is parasitic in the green bamboo (Bamusa textilis McCLURE) and Damjuk (Phyllostachys nigra var. Henonis STAPF). It is a mass of clumps of latex from the wounds of bamboo bees. Polygonal masses, the surface of which is milky white, off-white or off-white, light and easily crushed. When water is added, bubbles are formed but do not melt. Milky white, large, bright, and absorbent is good. It is produced in tropical and subtropical regions of southern China. This herb is antipyretic, jinkyung, soothing, expectorant, fever newlywed island language, pediatric curiosity, and stroke. It is used for sound and epilepsy, and the dose is stated to be taken by boiling 4-10 g per day or as a pill or powder.

The present inventors have conducted studies on fragranced HIV-1 substances in various kinds of natural products. A surprising fact with one activity was found to complete the present invention.

Accordingly, an object of the present invention relates to the use of the cheonnyeongsul, its extract and fractions fractionated from the extract for the treatment of AIDS and the use of immune fortification.

It is another object of the present invention to prepare a pharmaceutical preparation by the method of preparing a conventional pharmaceutical preparation together with excipients which are used as an active ingredient and containing the cheonnyeong sulfur, its extract and fractions fractionated from the extract. It is to provide a pharmaceutical formulation formulated.

Another object of the present invention is to prepare a pharmaceutical preparation by preparing a method of preparing a pharmaceutical preparation together with the excipients commonly used as pharmaceutically acceptable additives containing the sulfuric acid, its extract and fractions fractionated from the extract as an active ingredient. It provides a manufacturing method for manufacturing.

Extraction and fractionation of the sulfuric acid sulfur of the present invention is carried out by the following method.

Example 1

6.5 liters of re-distilled water (DDW) was added to 250 g of Korean solar sulfur and extracted using an electronic shaker for 2 hours and 30 minutes, filtered and dried to obtain 1.53 g of extract. 99% ethanol was added to the water extract in a ratio of 1: 2, and the sample was completely dissolved using a fraction funnel, followed by overnight at 4 ° C. The resulting precipitate and the supernatant were centrifuged at 15,000 rpm for 30 minutes with a 4 ° C refrigeration centrifuge, filtered with Whatman Filter No. 1, evaporated and dried to obtain 0.752 g of a precipitate and 0.573 g of a dried supernatant. Each sample obtained from the precipitate and the dried supernatant was fractionated in the following manner and the process is illustrated in the following scheme 1 and scheme 2.

Methanol was added to the dried sample and extracted overnight with stirring with a magnetic stirrer. The obtained methanol extract and the insoluble residue were centrifuged at 4 ° C. and 15,000 rpm for 30 minutes, filtered through Whatman filter paper No. 1, and the filtrate was evaporated to dryness. The dried methanol extract was dissolved in a small amount of DDW, centrifuged again by adding acetonitrile until a precipitate was formed, and the filtrate was separated into a supernatant and a precipitate and dried to obtain fractions B and C. The dried insoluble residue was washed with acetone for 30 minutes and then dried by centrifugation, and then dissolved in a small amount of DDW and added with acetonitrile until a precipitate was formed. The mixture was centrifuged and filtered to separate the supernatant and the precipitate, and then dried. The dried supernatant was defined as fraction D. The dried precipitate was washed with tetrahydrofuran (THF) and dimethyl formumamide (DMF) and then dried by centrifugation and filtration. Dimethyl sulfoxide (DMSO) was added to the dried sample and extracted by stirring overnight using a magnetic stirrer. The resulting DMSO extract and insoluble residue were centrifuged, filtered and dried. The dried DMSO extract was designated as fraction E, the dried insoluble residue was designated as fraction F, and anti-HIV-1 activity was investigated for these substances.

These processes are shown in the following scheme 1 and scheme 2.

Experimental Example 1

1. Anti HIV-1 Activity Test

1) Preparation of Susceptible Cells:

To detect in vitro anti-HIV-1 activity, MT-4 cells from a special pathogen laboratory in the National Institute of Health, Japan were passaged and used. MT-4 is a human T cell line containing HTLV-1, containing 10% fetal calf serum (FCS), penicillin (100 units / ml), streptomycin (100 μg / ml), and 2 mM L-glutamine per ml. The culture was incubated at 37 ° C., 5% CO using RPMI-1640 medium. Viability of the cells was confirmed by trypan blue dye exclusion method, and passaged once every 3-4 days 2.0-3.0X10 per ml Maintained as dog cells and used.

In addition, MOLT-4 and MOLT-4 / IIIB cells, which are human T cell lines similar to MT-4 cells distributed in the same laboratory, were used to elucidate the mechanism of action of the active substances. Is a cell line producing HIV-1, and the culture conditions are as described above.

2) Preparation of the virus:

HIV-1 stores the culture supernatant and cells of H9 / HTLV-III cells (ATCC No. CRL-8543) producing HTLV-III at a temperature below -70 ° C and repeats two to three freezes and thaws. The cells were completely destroyed and centrifuged for 30 minutes at 5,000 rpm in a 4 ° C refrigerated centrifuge. After harvesting the upper layer was dispensed 0.5ml each in a storage container of 1ml capacity and stored in -70 ℃ deep freezer (REVCO, ULT-1685) and taken out whenever necessary. Virus titer was determined by observing the cytopathic effect (CPE) formed in MT-4 cells.

3) HIV-1 Infection and Sample Activity Assessment:

MT × 4 cells 1.0 × 10 per well in 96 well cell culture vessels After incubation with dogs, 50 μl of 500TCID50 HIV-1 was added to each well, and then 100 μl of each sample was incubated for 6 days in a moisturizing incubator maintained at 37 ° C. and 5% CO. The anti-HIV-1 activity of the samples was evaluated using MTT cell proliferation kit I, which measures viable cells by the difference in absorbance at 550/690 nm, and 2,3-azidothymidine (AZT, Sigma) as a control drug. ) And dextrn sulphate (Pharmacia, MW 500,000) were used. In addition, self-efficacy evaluation was conducted for reference compounds known to exhibit anti-HIV-1 activity.

4) Determination of Cytotoxicity of Samples on MT-4 Cells:

Cytotoxicity by sample was 1.0 × 10 per well by diluting the sample by concentration. MT-4 cells were added to 96 well plates, each cultured for 6 days in 37 ℃, 5% CO moisturizing incubator and 50% cytotoxic dose (CD) was calculated by MTT assay was used in this experiment.

5) 3- (4,5-dimethylthiazol-2-yl) -2,5-diphenyl tetrazolium bromide (MTT) assay: The MTT assay used a commercially available MTT cell proliferation kit I (Boheringer Mannheim). 100 μl of the cell supernatant incubated for 6 days after diluting the sample and virus diluted by concentration in a 96well plate, and 10μl of MTT labeling reagent (final concentration 0.5mg / ml) was added to each well. After shaking gently using a microshaker, the mixture was allowed to stand for 4 hours in a 37 ° C, 5% CO moisturizing incubator. 100 μl of solubilization solution (SDS, 10% in HCL, 0.01 mol / l) was added to each well to completely dissolve formazan (formazan), a dark blue crystal that had been formed. Then, ELISA reader (SLT, ERA- 400) was measured for absorbance (test wavelength 550nm, reference wavelength 690nm). The anti-HIV-1 activity of the sample was determined by measuring the degree of inhibition based on the absorbance of the HIV-1 infected and non-infected control cell groups.

6) HIV-1 p24 antigen detection ELISA:

1.0 × 10 per well After inoculation of 500 TCID50 HIV-1 to MT-4 cells, each sample was added so that the final volume was 200 μl. After culturing for 6 days in a humidified incubator at 37 ° C and 5% CO, the culture supernatant of the specimen was taken by date to measure the amount of HIV-1 p24 antigen using the HIV-1 p24 core profile ELISA kit (Du Pont). It was.

30 μl of 5% Triton X-100 was added to 270 μl of the culture supernatant diluted 100-fold and used as a sample. Each well of the anti-HIV-1 p24 coated microplate was washed 10 times with a washing solution, and 200 μl of sample, positive and negative controls were added before the microplate was dried. Positive controls resulted in concentrations of HIV-1 p24 of 100, 50, 25 and 12.5 pg per ml for standardization. The plate was covered with a sealer, reacted at 37 ± 1 ° C. for 2 hours, washed 10 times, 100 μl of detector antibody was added, the plate was resealed, and reacted at 37 ± 1 ° C. for 1 hour. After washing 10 times, add 100μl of streptavidin-HRP (horseradish peroxidase), seal the plate, react for 15 minutes at 37 ± 1 ℃, and then wash 10 times to add 100μl of substrate (orthophenylene diamine, OPD). It was left to stand at room temperature for a minute.

100 μl of reaction stopper (4N sulfuric acid) was added thereto, and the absorbance was measured at 492 nm (test wavelength) and 620 nm (reference wavelength), respectively, using an ELISA reader (SLT, ERA-400).

7) Mechanism of action of active substance on HIV-1:

(1) Use of MT-4 Cells:

To determine the mechanism by which extracts exhibit anti-HIV-1 activity in cells, inject MT-4 cells into 15 ml conical tube, inoculate HIV-1 with 0.05 multiplicity of infection (MOI), and shake them evenly at 15-minute intervals. The mixture was allowed to stand at 37 ° C. in a 5% CO moisturizing evaporator for 1 hour. Centrifuge at 1,200 rpm for 15 minutes, discard the upper layer, centrifuge three times with medium without FCS, suspend in culture medium, and then 1 × 10 per well. Cells were aliquoted into 96 well plates. Active substances diluted by concentration were added to each other, and then cultured for 6 days in a 37 ° C., 5% CO moisturizing incubator, followed by MTT assay.

In addition, in order to determine whether the extract inhibits the direct action of HIV-1 or the adsorption of HIV-1 to the cell surface, the mixture is mixed with HIV-1 and diluted active material for 1 hour and then reacted for 96well. 1 × 10 per well on the plate MT-4 cells cultured in dogs were incubated for 6 days in a 37 ° C., 5% CO moisturizing incubator and evaluated for their mechanism of action by MTT assay.

(2) Use of MOLT-4 and MOLT-4 / IIIB cells

5 × 10 per well in a 96well plate to determine whether the extract inhibits the adsorption of HIV-1 to the cell surface The same number of MOLT-4 / IIIB cells and the same number of MOLT-4 cells were dispensed at the same time, and the extracts diluted by concentration were added. After incubation for 1 day at 37 ℃, CO moisturizing incubator was evaluated by the inverted microscope (synytium) formation to evaluate the mechanism of action.

8) Combined Test of Extract Quality and AZT:

10 × 10 per 96 well 4 MT-4 cells, added 500TCID50 of HIV-1, followed by 0.1ED50 (0.000.μM), 1ED50 (0.001μM), 10ED50 (0.01μM) AZT and 0.1ED5O (4μM), 1ED50 (40μM), 10ED50 (400μG) extract NO. After 142-2A was added to each sample alone and combined, the sample was incubated for 6 days in a moisturizing incubator maintained at 5 ° C at 37 ° C, and the absorbance was measured by MTT assay to evaluate synergism and antagonism. It was.

II. Anti HIV-1 activity test result

1) Anti-HIV-1 Activity of Hot Water Extracted Sample No. 142:

The anti-HIV-1 activity of sample No. 142 obtained by hot water extraction was very strong in vitro. The precipitate (No. 142-2A) of the material obtained by mixing this material with 99% ethanol, centrifugation and filtration was divided into four fractions (No. 142-2B, D, E, and F) and the supernatant (No. 142-). 3A) was separated and purified into five fractions (No. 142-3B, C, D, E and F) and the results of the anti-HIV-1 activity test are shown in Table 1. As confirmed in this table, No. Anti-HIV-1 efficacy was observed in all fractions except 142-3B. Among them, No. 142-2A, 142-3E and 142-3F are Nos. It showed higher anti-HIV-1 activity than 142. Pre-fractionation substance No. 142 was CD50 = 663.66μg / ml, ED50 = 52.15μg / ml, SI = 12.73, but CD50 = 629.03μg / ml, ED50 = 30.64μg / ml, SI = 20.53 of substance 142-2A after fractionation. Decreased, resulting in high anti-HIV-1 activity. Also no. 142-3E showed CD50 = 697.60μg / ml, ED50 = 14.22μg / ml, SI = 49.06, which increased CD50, decreased cytotoxicity, and decreased ED, showing the strongest anti-HIV-1 activity among fractions. CD50 of 142-3F, CD50 = 864.86μg / ml, ED50 = 43.41μg / ml, SI = 19.92 increased CD50 and decreased cytotoxicity, and ED50 decreased, resulting in high anti-HIV-1 activity. Therefore, sample No. 142 has been suggested as a candidate for anti-HIV-1 activity.

As can be seen from this result, it is judged that the water extracts of the yellow snapper, and most of the fractions fractionated therefrom, have anti-HIV-1 activity. The fact that several fractions have anti-HIV-1 activity is considered to have several types of HIV-1 activators. Fractionants 142-2A, 142-3E, and 142-3F are considered to have high activity.

FIG. 1 is a graph showing MTT foramazan production in MT-4 cells cultures treated with No.142 extract. FIG. 2 is a graph showing MTT foramazan production in MT-4 cells cultures treated with No.142-2A extract. FIG. 3 is a graph showing MTT foramazan production in MT-4 cells cultures treated with No.142-3E extract. FIG. 4 is MTT foramazan in MT-4 cells cultures treated with No.142-3F extract. Graph showing production.

As identified in these graphs, fraction No. It can be seen that 142, 142-2A, 142-3E and 142-3F have high anti-HIV-1 activity.

2) Sample No. 142 HIV-1 Proliferation Inhibition Test by Fractional Substances:

(1) HIV-1 p24 antigen detection ELISA:

Extract Quality No. The inhibitory effect of 142 fractions on the proliferation of HIV-1 in MT-4 cells was confirmed by date using HIV-1 p24 antigen detection ELISA, a viral antigen detection method. -(test sample OD / virus control OD) x 100].

Extract Quality Fraction No. 142-2A was divided into 7 concentrations (500-7.8125μg / ml), and the control agent AZT was used at 0.01μM / ml to react with HIV-1 in MT-4 cells, respectively. The solution was diluted 100-fold and the amount of HIV-1 p24 antigen was measured for absorbance at 492/620 nm. The results are shown in Table 2 and FIG.

The absorbance of the control cell group and the control virus group after 4 days of culture was 0.027 and 2.629, respectively, and after 6 days of culture, the absorbance was 0.007 and 2.650. After 4 days of AZT control, the absorbance was 0.059 and the HIV-1 inhibition rate was 97.8%. After 6 days, the absorbance was 2.070 and the inhibition rate was 21.9%. Therefore, up to 4 days after culture, HIV-1 proliferation was almost suppressed by AZT, but by day 6, it was found that the effect of AZT on HIV-1 was greatly reduced. No. 142-2A inhibited HIV-1 similarly to AZT with 98% inhibition rate of 98% at 500-65.2μg / ml between farms. Some antiviral activity was observed at 31.25 and 15.625μg / ml. Unlike AZT, however, No.142-2A showed more than 99.0% HIV-1 proliferation inhibitory activity at the concentration range of 500-250 μg / ml on day 6, and the ingibition rate was 88.4% and 52.1 at 125 and 62.5 μg / ml, respectively. As a percentage, No. 142-2A inhibited HIV-1 proliferation more strongly than AZT.

FIG. 5 shows No. 1 for HIV-1 p24 antigen expression in MT-4 cells. It is a graph showing the inhibitory effect according to the culture days of 142-2A and AZT.

Also no. Incubate 142-3E at 7 concentrations (500-7.8125μg / ml) in the same manner as above, collect supernatant after 4, 5, 6, 7 days, and dilute 100 times to measure the amount of HIV-1 p24 antigen. It was. The results are shown in Table 3 and FIG.

The absorbance of the control cell group measured during this period was 0.017-0.028, and the absorbance of the control virus group was more than 3.0. In the concentration range of 500-62.5 μg / ml, the absorbance was 0.074-0.090 and almost no HIV-1 p24 antigen was detected even after 4-6 days of culture. On the 7th day, the antigen detection was started at 62.5 μg / ml with the absorbance of 0.647. In contrast, HIV-1 p24 antigen was detected on day 4 of culture at concentrations below 31 μg / ml.

FIG. 6 shows No. 1 for HIV-1 proliferation inhibition in MT-4 cells by HIV-1 p24 Antigen detection ELISA. Graph showing the effect of 142-3E.

As can be seen from the above results, the fractions of the present invention are compared at higher concentrations than AZT, but this is inevitable due to the nature of natural products, but in terms of actual effects, the effect is superior to AZT and the effect is persistent. have.

It is thought that the drug of the present invention can bring a better effect when used in combination with the existing anti-AIDS agent AZT and the like.

(2) Comparison of MTT assay and HIV-1 p24 antigen detection ELISA:

After 6 days of culture, HIV-1 proliferation inhibitory ability was compared with MTT assay and HIV-1 p24 antigen detection ELISA. The results are shown in Table 4 and FIG. For the MTT assay, which indirectly evaluates anti-HIV-1 activity by quantitatively measuring cells that do not die of HIV-1, no. Although 142-2A showed the strongest absorbance around 62.5μg / ml than other concentrations, it showed inhibition rate of 65.4-70.5% against HIV-1, but 500 in ELISA that directly measures the amount of HIV-1 p24 antigen. -In the concentration range of 250μg / ml, more than 99.0% of virus proliferation inhibitory ability was observed. In other words, the high concentration (500-250μg / ml) of No 142-2A acting on HIV-1 was very suppressed by HIV-1 p24 antigen and showed a little cytotoxicity due to the small amount of living cells. At 125 to 31.25 μg / ml, the degree of inhibition of HIV-1 p24 antigen decreased gradually, while the amount of living cells increased and then decreased again by HIV-1 infection.

FIG. 7 shows No. 1 for the inhibition of HIV-1 proliferation in MT-4 cells by MTT assay and HIV-1 p24 antigen detection detection ELISA. Graph showing the effect of 142-2A.

Also no. HIV-1 proliferation inhibitory effect by 142-3E is shown in Table 5. After 6 days of culture, the absorbance by MTT assay in the concentration range of 250-625 μg / ml was 0.946-1.167, similar to the absorbance 1.043 of the control cell group, but the amount of HIV-1 p24 antigen was 62.5 μg / Absorption was hardly detected at 0.065-0.090 above ml, and at concentrations below that, a very large amount of HIV-1 p24 antigen was measured at an absorbance of 3.0 or higher irrespective of the amount of viable cells.

Therefore, the MTT assay was suitable for detecting the anti-HIV-1 activity of the drug, but it was not suitable for measuring the quantitative decrease of the virus. Antigen detection ELISA method was suitable.

3) Mechanism of action of extract on HIV-1:

(1) Use of MT-4 Cells:

No. extract quality The results of experiments to determine at what stage 142 inhibits HIV-1 replication are shown in Figures 8 and 9. Sample No. When 142-2A and HIV-1 were reacted for 1 hour in advance and inoculated into MT-4 cells, the anti-HIV-1 activity was evaluated by MTT assay 6 days after culture. CD = 918.33 μg / ml, ED = 11.07 μg / ml , SI = 82,95 showed strong anti-HIV-1 activity, but when MT-1 cells were infected with HIV-1 for 1 hour and anti-HIV-1 activity was assessed by MTT assay, CD = 1,000.00μg / ml Did not show anti-HIV-1 activity at all. As a result, Sample No. 142 was identified as a substance that directly acts on HIV-1 or inhibits the adsorption of HIV-1 to MT-4 cells.

FIG. 8 is a graph showing MTT formazan production in MT-4 cells after 1 hour pre-reaction with No 142-2A and HIV-1. FIG. 9 shows pre-reaction of MT-4 cells with HIV-1 for 1 hour. This is a graph showing MTT formazan production in MT-4 cells.

(2) Use of MOLT-4 and MOLT-4 / IIIB cells:

NO. Synthium formation of MOLT-4 and MOLT-4 / IIIB cells with reference to AZT and dextran sulphate, a mechanism of action previously identified, to determine at what stage 142 inhibits HIV-1 replication Evaluation was made through observation. The results are shown in Table 6. As shown in Table 6, AZT forms neocytium at all concentrations (10-0.000128 μg / ml) at 8 concentrations that are not cytotoxic, and as previously known, HIV-1 does not inhibit adsorption on the cell surface. It was confirmed. Dextran sulphate did not form synthium at 1,000-8 μg / ml, but did not form synthium at lower concentrations (1.6-0.000128 µg / m), thus inhibiting HIV-1 cell surface adsorption. . Sample No. No. 142 fraction substance. 142-2A inhibited synthium formation by the same mechanism as dextran sulphate at the concentration of 1.000-125µg / ml, and synthium was formed in the experimental section below (62.5-7.8125µg / ml). Therefore, No. Like dextran sulphate, 142-2A was identified as a substance that inhibits HIV-1 from adsorbing on the surface of cells.

4) Combination of Drugs for Anti-HIV-1 Activity Assessment:

AZT and Sample No. 142-2A was mixed to show stronger anti-HIV-1 activity than the respective drugs by MTT assay, and the concentration of AZT was 0.0001, 0.001, 0.01 μM / ml, No. The concentration of 142-2A was set to 4, 40, 400 μg / ml.

No. mixed with an AZT concentration of 0.0001 μM The absorbances at 142-2A concentrations of 4, 40 and 400 μg / ml were 0.385, 0.633 and 0.704, respectively. The absorbance of 142-2A was much higher than 0.282, 0.355, and 0.506, indicating a synergistic effect due to the mixing of the two agents. Whereas the other two concentrations of AZT were mixed with 0.001, 0.01 μM / ml. 142-2A showed much lower absorbances than 0.989 and 0.795 when using AZT alone. Antagonism in which the efficacy of AZT is inhibited by 142-2A was observed. The results are shown in Table 7.

As evident from the above experiments, the drug of the present invention was found to be highly preferable when used as a combination because of the synergistic effect when combined with the existing AIDS therapeutic agents AZT and dextran sulphate at low concentrations.

Experimental Example 2

Sample No. 142 Acute Toxicity Tests:

1) Test Method:

Five to six weeks old ICR mice and mormites were used as experimental animals. In order to measure the acute toxicity of 142-3E, samples were orally administered at a predetermined amount and observed for the death of experimental animals for 7 days, and LD50 (50% of lethal dose) was obtained.

2) Test result:

Sample No. When 142-4E was observed for 4 days (10mg / 0.2ml, 5mg / 0.2ml, 2.5mg / 0.2ml, 1.25mg / 0.2ml) after oral administration, it showed weight loss and other symptoms compared to the normal control group. And survived healthy. Sample No. used in this experiment Since the maximum dose of 142-3E was 10 mg / ml, it was unreasonable because of the limitation of the orally administered sample. In addition, at the highest oral dose (10mg / 0.2ml), it was difficult to calculate the LD value because the mouse did not die and showed no other symptoms.

It is sample no. When 142-3E was observed for 7 days after oral administration of 10 mg / 0.5ml, the mice survived without weight loss and other symptoms as compared with the normal control group. Therefore, sample No. 142-3E and other fractions are considered nontoxic.

As can be seen in the above experimental example, the sheet of sulfur of the present invention, the water extract material, the fraction material fractionated it can be used as a drug for the treatment of AIDS (AIDS) and immunopotentiation. The amount used may be 1-10 g per day, if the herbal medicine itself. For water extracts, 100 mg-1000 mg per day can be used. In the case of fractions, 10 mg-500 mg per day can be used. These daily doses can be used one to several times daily. The amount of use can be increased or decreased depending on the age, sex and severity of the patient.

The sulfuric acid of the present invention, the water extract material and the fraction material thereof are used alone or in combination with the pharmaceutically conventionally used excipients in the form of pharmaceutically-used phosphates, tablets, It may be used in the form of preparations such as capsules, injections, solutions and ointments.

Next, a formulation example is illustrated.

Preparation Example 1

[Production of powder]

2g of India's yellow powder

1g lactose

The above ingredients are mixed and filled in airtight cloth to prepare a powder.

Preparation Example 2

[Production of powder]

No. 142 Extract 500mg

Corn Starch 100mg

Lactose 100mg

Talc 10mg

The above ingredients are mixed and filled in a bubble to prepare a powder.

Preparation Example 3

[Production of Tablets]

No. 142-2A Fractionate 100mg

Corn Starch 100mg

Lactose 100mg

2 mg magnesium stearate

After mixing the above components, tablets are prepared by tableting according to a conventional method for preparing tablets.

Preparation Example 4

[Production of Tablets]

No. 142-3E Fractional substance 50mg

Lactose 100mg

Corn Starch 50mg

1 mg magnesium stearate

After mixing the above components, tablets are prepared by tableting according to a conventional method for preparing tablets.

Preparation Example 5

[Production of Tablets]

No. 142-3F Fractionate 50mg

Lactose 50mg

1 mg magnesium stearate

After mixing the above components is compressed according to the conventional method for producing a capsule to fill the gelatin capsule to prepare a capsule.

Preparation Example 6

[Production of Capsule]

No. 142 Extract 100mg

Lactose 100mg

Corn Starch 100mg

1 mg magnesium stearate

After the above ingredients are mixed, the capsules are prepared by filling the gelatin capsules according to a conventional method for preparing capsules.

Preparation Example 7

[Production of injection preparation]

No. 142-2A Fractional substance 10mg

Appropriate sterile distilled water for injection

pH adjuster

According to the conventional method for preparing an injection, the active ingredient is dissolved in distilled water for injection, the pH is adjusted to about 7.5, the whole is adjusted to 2 ml with distilled water for injection, and then filled into a 2 ml ampoule and sterilized to prepare an injection.

Preparation Example 8

[Production of Injection]

No. 142-3F Fractionate 10mg

Suitable amount of distilled water for injection

pH adjuster

According to the conventional method for preparing an injection, the active ingredient is dissolved in distilled water for injection, the pH is adjusted to about 7.5, the whole is adjusted to 2 ml with distilled water for injection, and then filled into a 2 ml ampoule and sterilized to prepare an injection.

Preparation Example 9

Preparation of Liquid

No. 142 Extract 1g

10 g of isomerized sugar

10g per book

Lemon flavor

Purified water

According to the conventional method for preparing a liquid solution, each component is added to the purified water, dissolved, lemon flavor is added, and then purified water is added to adjust the whole amount to 100 ml, and then filled into a brown bottle, and then sterilized to prepare a liquid solution.

Claims (4)

Chemella (Bambusae Concretio Silicea), its water extract (Code No. 142) or fractions (Code Nos. 142-2A, 142-3E and 142-3F) fractionated from this water extract are prepared A preparation for the treatment of AIDS (AIDS), which is formulated in the form of a conventionally used pharmaceutical form with the excipients used and conventional pharmaceutical methods. The agent for treating AIDS according to claim 1, wherein the pharmaceutical form is a pharmaceutical form selected from powders, tablets, capsules, solutions, injections, ointments, and syrups. The method according to claim 1, wherein the biscuit (Bambusae Concretio Silicea), its water extract (Code No. 142) or the fraction material obtained by fractionating the water extract (Code Nos. 142-2A, 142-3E and 142-3F) Immunopotentiating formulations formulated in the form of a conventional pharmaceutically used excipient and a conventional pharmaceutical used in the usual pharmaceutical method. The preparation for immunopotentiation according to claim 3, wherein the pharmaceutical form is used in the form of a powder, tablet, capsule, liquid, injection, ointment, or syrup.