JPH07206668A - Therapeutic agent for human immune deficiency virus infectious disease - Google Patents

Abstract

PURPOSE:To obtain a therapeutic agent for a human immune deficiency virus infectious disease having low toxicity and no fear of adverse effect, comprising vinigrol as an active ingredient. CONSTITUTION:This therapeutic agent for a human immune deficiency virus infectious disease comprises a vinigrol (FR-900478 substance) a fermentation product of a fungus of the genus Virgaria of Fungi Imperfecti and/or its salt as an active ingredient. Vinigrol has excellent treating effect on HIV infectious disease and low acute and cytotoxicity. Vinigrol can be pharmaceutically prepared into a form of oral administration such as tablet, capsule, granule, powder or syrup or into a form of parenteral administration such as injection, intravenous drip and suppository and a dose is 0.1-1,000mg per time.

Description

Detailed Description of the Invention

[0001]

BACKGROUND OF THE INVENTION The present invention relates to vinigrole (Vi
nigrol) and / or a pharmacologically acceptable salt thereof as an active ingredient, to a therapeutic agent for human immunodeficiency virus infection.

[0002]

BACKGROUND OF THE INVENTION Human immunodeficiency virus infection is a disease in which human immunodeficiency virus significantly impairs the immunocompetence of a host, and indirectly causes infection by various pathogenic bacteria, resulting in death of the patient.

Therefore, research and development of preventive and therapeutic agents for human immunodeficiency virus infection have been carried out, and to date, azidothymidine (AZT) (H) has been used as a typical therapeutic agent for inhibiting the growth of human immunodeficiency virus. Mituya e
t al. , Pro. Natl. Acad. Sci. U
SA, 82 , 7096-7100 (1985)) has been developed. In addition, as a typical therapeutic drug that inhibits the adsorption of human immunodeficiency virus to target cells, dextran sulfate (H. Mituya et al., Sci) is used.
ence, 240 , 646-649 (1988)) and soluble CD4 (N. Watanabe et a.
l. , Nature, 337 , 267-270 (198).
9)) etc. have been developed.

However, although these known therapeutic agents have certain effects, they are not sufficiently satisfactory in terms of efficacy and safety, and there is a strong demand in the art for the development of superior therapeutic agents. ing.

[0005]

DISCLOSURE OF THE INVENTION The present invention is directed to the present state of the art in which the development of novel and superior therapeutic agents as described above is strongly demanded as the number of patients with acquired immunodeficiency syndrome (AIDS) increases. In view of this, the present invention has been made in view of the purpose of developing a preventive and therapeutic agent for a novel human immunodeficiency virus infectious disease which has hitherto been unknown.

[0006]

[Means for Solving the Problems] As a result of intensive research on various substances to achieve the above-mentioned object, it was found that vinigrole inhibits infection of human immunodeficiency virus, and further research is conducted based on this new finding. The present invention has been completed.

Vinigrol, which is an active ingredient compound in the present invention, is FR-90047.
It is also called 8 substances, and is an incomplete bacterium Virgaria nigra F-5408 (Virgaria nigr
a F-5408, FERMP-8753), it has been clarified by the inventors of the present invention that the substance is one of the substances produced from F-5408, FERMP-8753).
0).

[0008] The present invention comprises vinigrole and / or a pharmaceutically acceptable salt thereof as an active ingredient. Examples of such salts include alkali metals such as sodium and potassium; lysine and arginine. Examples thereof include salts with basic amino acids; and the like. Furthermore, the vinigrole of the present invention can be used in the form of a non-toxic pharmaceutically acceptable acid addition salt. Examples of such acid addition salts include inorganic acids such as hydrochloric acid, sulfuric acid, nitric acid and phosphoric acid; acetic acid, succinic acid, maleic acid, fumaric acid, malic acid, glutamic acid, aspartic acid, p-toluenesulfonic acid, methane. An acid addition salt with an organic acid such as sulfonic acid;

However, the pharmacologically acceptable salt of the present invention is not limited to these.

The vinigrole used in the present invention exhibits excellent anti-human immunodeficiency virus properties. The activity is measured by R. It can be measured according to the method of Pauwel et al. (R. Pauwel et al., J. Virol).
optical Method, 20 , 309-321
(1988)). That is, human immunodeficiency virus-infected MT-4 cells (human lymphoid cells,
The same applies hereinafter) and human immunodeficiency virus-uninfected MT-4 cells to which a test compound has been added are respectively cultured. Similarly, human immunodeficiency virus-infected MT-4 cells without a sample compound and human immunodeficiency virus-uninfected MT-4 cells without a sample compound are respectively cultured. Then, the MTT method (LM Green et al., J. Immun.
olo. Method, 70 , 257-268 (198).
4)) to measure the viable cells to determine the cytotoxic activity of human immunodeficiency virus. Thereby, the anti-human immunodeficiency virus activity of the sample compound can be measured.

Vinigrole (salt) is administered in various forms. Examples of the dosage form include oral administration by tablets, capsules, granules, powders, syrups and the like, or parenteral administration by injections (intravenous, intramuscular, subcutaneous), infusions, suppositories, etc. . These various preparations are usually used in the technical field of pharmaceutical preparation such as excipients, binders, disintegrants, lubricants, flavoring agents, solubilizers, suspension agents, coating agents, etc. It is possible to formulate with known auxiliary agents. The dose varies depending on symptoms, age, body weight, administration method, dosage form, etc., but is usually about 0.1 mg to 1, once per adult.
000 mg can be administered.

The active ingredient compound of the present invention exhibits excellent anti-human immunodeficiency virus (anti-HIV) activity, as is apparent from the examples described below. The details of its mechanism of action must be investigated in the future, but at present, it is assumed that this compound directly acts on HIV.

The active ingredient compounds of the present invention are excellent anti-H compounds.
In addition to having IV activity, the LD ₅₀ was 23 mg / kg (i.d.) in an acute toxicity test using ddY mice.
p. ), And p. o. In the case of, LD ₅₀ is> 10
It was 0 mg / kg, and it was confirmed that it was also excellent in safety.

The embodiments of the present invention will be described in detail below.

[0015]

Example 1 Anti-human immunodeficiency virus activity was measured by a conventional method (R. Pauwelet al., J.
Virological Method, 20 , 309
-321 (1988)).

That is, MT-4 cells in the logarithmic growth phase were centrifuged at 150 × g for 5 minutes, and human immunodeficiency virus type 1 (HTLV-IIIb strain) was added to the resulting cell pellet as M.
The cells were infected at a concentration of OI: 0.01 at 37 ° C for 1 hour. Thereafter, human immunodeficiency virus-infected MT-4 cells were obtained by centrifugation in RPMI-1640 medium supplemented with 10% fetal bovine serum (hereinafter referred to as "serum medium") and washing. Human immunodeficiency virus-infected MT-4 cells and human immunodeficiency virus-uninfected MT-4 cells were each 2.5.
The cells were suspended in a serum medium at a density of × 10 ⁵ cells / ml.

In a 96-well plastic microtiter plate, 100 μl of each pre-diluted sample compound solution (suspended in serum medium) was added to each well, and then 100 μl of each of the above cells was added to each well. Static culture was carried out at 37 ° C. for 5 days in the presence of 5% carbon dioxide. The analyte compound has a pure concentration of 1000, 200,
40, 8, 1.6, 0.32, 0.064, 0.012
The test was conducted at 8, 0.0026 μM.

Similarly, human immunodeficiency virus-infected MT-4 cells containing no test compound and human immunodeficiency virus-non-infected MT-4 cells containing no test compound were prepared.
It was cultured at 37 ° C. for 5 days in a CO ₂ incubator.

After completion of the culture, MTT (3- (4,5-di
meththiazol-2-yl) -2,5-di
phenyltetrazolium bromid
e) The live cells were measured based on the method (LM Green)
et al. J. Immunolo. Metho
d, 70 , 257-268 (1984)).

The antiviral activity is a concentration (EC ₅₀ : 50% Ef) that suppresses and protects cell damage caused by HIV infection by 50%.
It was shown as a positive Concentration). That is, the cytotoxic activity of human immunodeficiency virus-infected MT-4 cells without addition of a sample compound was set to 100%, and human immunodeficiency virus-uninfected M without a sample compound was added.
When the cytotoxic activity of T-4 cells was set to 0%, the concentration (EC ₅₀ ) of the sample that suppressed the cytotoxic activity of MT-4 cells infected with human immunodeficiency virus by 50% was determined and shown at this concentration.

The cytotoxic activity of the test compound is a concentration at which the growth of human immunodeficiency virus-uninfected MT-4 cells is suppressed by 50%, that is, a 50% cytotoxic concentration due to the test compound (CC ₅₀ : 50% Cytotoxic Conceal).
).

Furthermore, the selection coefficient (SI: Selective Index) of human immunodeficiency virus activity is CC.
Calculated as ₅₀ / EC ₅₀ value.

The measurement results obtained as described above are shown in Table 1 below, and EC ₅₀ , EC ₉₀ , CC ₅₀ and SI values are shown in Table 2 below.

[0024]

[Table 1]

[0025]

[Table 2]

Table 3 below shows a comparison of the effects of dextran sulfate and azidothymidine (AZT), which are anti-HIV agents conventionally used in order to show excellent anti-HIV activity of vinigrol.

[0027]

[Table 3]

From these results, the following things became clear. First, the control human immunodeficiency virus-infected MT-4 cells were all killed by the human immunodeficiency virus, but vinigrole increased the survival rate of human immunodeficiency virus-infected cells depending on the concentration thereof. That is, as is clear from the EC ₅₀ values in the table, it showed a remarkable anti-human immunodeficiency virus activity.

Furthermore, the cytotoxic activity of Vinigrole on MT-4 cells not infected with human immunodeficiency virus (C
C ₅₀₎ was weak.

Further, as is clear from the comparison data with the above-mentioned known anti-HIV agents, vinigrole is anti-human in comparison with the reverse transcriptase inhibitor azidothymidine and the human immunodeficiency virus adsorption inhibitor dextran sulfate. Weaker than azidothymidine in immunodeficiency virus activity,
It was stronger than dextran sulfate. However, the cytotoxic activity (CC ₅₀ ) was weaker than that of azidothymidine.

From the above results, it was revealed that vinigrole exhibits a strong anti-human immunodeficiency virus activity higher than that of dextran sulfate and has a high selectivity coefficient.

[0032]

INDUSTRIAL APPLICABILITY Vinigrol, which is the active ingredient compound of the present invention, has not only an excellent therapeutic effect on HIV infection, but also has a great feature of low acute toxicity and cytotoxicity.

Therefore, the present invention can be advantageously used in the treatment and prevention of HIV infections without worrying about particular side effects.

Claims (1)

[Claims] 1. A therapeutic agent for human immunodeficiency virus infectious disease, which comprises Vinigrol and / or a pharmacologically acceptable salt thereof as an active ingredient.