JPH07179475A - Polyamine zinc complex and antiviral agent for acquired immune deficiency syndrome containing the same - Google Patents

Abstract

PURPOSE:To obtain a polyamine zinc complex useful as antiviral agent for acquired immune deficiency syndrome, having low toxicity and high activity against a cell infected with human immune deficiency virus. CONSTITUTION:This polyamine zinc complex of the formula. The compound of the formula, for example, is obtained by mixing and reacting a methanol solution of cyclam (1,4,8,11-tetraazacyclotetradecane) with a methanol solution of Zn(ClO4)2.6H2O at 60 deg.C.

Description

Detailed Description of the Invention

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a novel polyamine zinc complex which exhibits antiviral activity and is useful as a medicine, and an anti-acquired immunodeficiency syndrome virus agent containing the polyamine zinc complex as an active ingredient.

[0002]

2. Description of the Related Art Anti-acquired immunodeficiency syndrome is a disease prevalent among drug abusers such as narcotics and male homosexuals. Infection from blood products is also a problem. Normally, AIDS (Acquired I)
mmuno Deficiency Syndoro
me). When this disease is caused, the immune function is significantly reduced, and the rate of infectious diseases is high, and the rate of death due to infectious diseases is also extremely high.

Many proposals have been made for the mechanism of the onset of such AIDS. For example, AIDS
It is considered that a virus (HIV) infects CD-4 antigen-positive helper T cells, which are target cells, one after another, and repeats virus proliferation and cell destruction to cause immunodeficiency. However, much remains unknown about the mechanism of AIDS development.

Molecular biological research in this field includes
There are some remarkable things, and the characteristics of HIV are gradually becoming clear. As a result, HIV is characterized in that the viral gene structure is remarkably complicated as compared with other retroviruses, and that it is a virus that exhibits diversity in terms of functions such as infectious disease and proliferative ability.

Based on these findings, AIDS virus growth inhibition methods, infection prevention methods and the like have been proposed. A
The main inhibitors of IDS virus growth are nucleic acid derivatives and reverse transcriptase inhibitors.

The most well-studied one is A
There is ZT (azidothymidine). AZT selectively suppresses HIV growth even in a freshly infected system of MT-4 cells. In clinical trials, the efficacy was confirmed in a control experiment using placebo as a control. However, it is known that long-term administration has side effects such as bone marrow disorder, and there is an urgent need to develop a drug to replace it.

As described above, effective drugs against the AIDS virus are currently being developed and put into practical use.

However, an anti-AIDS virus agent having sufficient efficacy has not yet appeared, and there remains a problem of improving the therapeutic effect of the anti-AIDS virus or the side effect of administration, and the emergence of a drug to replace it. Is desired.

[0009]

Therefore, the object of the present invention is to provide a novel anti-AID having properties meeting the above objects.
To provide an S virus agent.

Another object of the present invention is to provide an anti-AIDS virus agent which has a strong antiviral effect and has extremely low side effects.

[0011]

These objects are achieved by the present invention having the following constitution.

(1) A polyamine zinc complex represented by the following structural formula 2.

[0013]

[Chemical 3]

(2) An anti-acquired immunodeficiency syndrome virus agent containing the polyamine zinc complex according to (1) as an active ingredient.

(3) An anti-acquired immunodeficiency syndrome virus agent containing a polyamine zinc complex represented by the following structural formula 4 as an active ingredient.

[0016]

[Chemical 4]

When the anti-AIDS virus agent of the present invention is made into an aqueous solution, it is chemically sufficiently stable in the pH range of 4-8.

The anti-AIDS virus agents according to the invention are usually represented by the structural formula 3 according to the invention in a solid or liquid carrier.
Alternatively, it contains at least one compound represented by Chemical formula 4. The compounds according to the invention can also be used in combination with known active substances.

According to the invention, the dosage form of the anti-AIDS viral agent can be of any suitable and / or conventional type. When the compound is administered by the oral, subcutaneous, intravenous and intraperitoneal routes, the carrier is a sterile liquid of distilled water or saline, especially water. Generally, the water carrier of an injectable aqueous solution is 10 mg / ml (1.
0%).

The compound according to the present invention can be administered in an oral dosage form, and is suitable as a dosage form for ointments, external eye drops and the like. However, the preferred oral dosage form is a tablet,
Capsules, powders, suspensions and the like. In these dosage forms, at least 1.0-3.0% of the total composition
(Weight) is preferred. The dose of the compound for an adult is usually 10 mg to 3000 mg per day.

Hereinafter, the present invention will be described in more detail with reference to examples.

[0022]

【Example】

(Example 1) Cyclam (1, 4, 8, 11-tetr
aazacyclotetradecane, 200m
g, methanol solution (5ml) was heated to 60 ° C. of 1.00 mmol), stirring Zn (ClO _{4) 2} · 6H
_A solution of ₂ O (390 mg, 1.05 mmol) in methanol (5 ml) is added dropwise over 10 minutes.

After further heating at 60 ° C. for 1 hour and then allowing to cool to room temperature, a colorless cyclam zinc complex (312 mg, 0.67 mmol, 67) having the structure shown in the above formula 3 is obtained.
%) Crystals were obtained.

The spectroscopic data of this support the structure of Formula 3 above.

[0025]

Infrared absorption: 3420, 3272, 293
0,2876,1474,1458,1429,129
6,1144,1109,1092,1022,98
0,966,939,893,873,627cm ^-1

¹ NMRinD ₂ O (δ): 1.58-1.
71 (2H, m), 1.87-1.95 (2H, m),
2.48 (4H, t), 2.71 (4H, quinte
t), 3.00-3.23 (8H, m)

Example 2 Cyclen (1, 4, 7, 1)
0-tetraazacyclotetradecane
e, 300 mg, 1.74 mmol) in ethanol solution (10 ml) was heated to 70 ° C., and Zn (C
_{lO 4) 2 · 6H 2 O} (650mg, 1.74mmol)
Of ethanol solution (5 ml) is added dropwise over 10 minutes.

After heating for 1 hour at 60 ° C. and then allowing to cool to room temperature, a colorless cyclen-zinc complex (monohydrate, 550 mg, 1.22 mmo) having the structure shown in Formula 4 above is obtained.
1, 70%) crystals were obtained.

The spectroscopic data of this support the structure of Formula 4 above.

[0031]

Infrared absorption: 3418, 3304, 317
5,2955,1481,1445,1375,128
0,1242,1143,1111,1010,96
5,902,864,806,627cm ^-1

¹ NMRinD ₂ O (δ): 2.69-2.
78 (8H, m), 2.90-2.98 (8H, m)

Example 3 Measurement of Anti-Human Immunodeficiency Virus (HIV) Growth Inhibitory Activity HIV growth inhibitory activity was measured according to the method of Nakamura et al. (Antiviral Research).
Research), 15, 113-124, (199
1)).

The measurement conditions are as follows.

Cell: MT-4 (HTLV-I-carr
ying cell line). Virus: HIV-1 (a type of HIV). The virus titer was 1 × 10 ⁶ TCID ₅₀ / ml.

Medium: RPMI-1640 (containing 10% fetal bovine serum, 100 IU / penicillin and 100 μg / ml streptomycin).

Culture conditions: MT-4 cells (1 × 10 ^{5 /} ml, 0.2 ml) and HI were placed in each well of a 24-well culture tray.
V-1 (2 × 10 ⁴ TCID _{50 /} ml, 0.2 ml) was added (MOI = 0.02), and the cells were cultured for 1 hour. After adsorbing the virus, the compounds according to the present invention (0.6 ml) each having the structure of the present invention diluted with a medium (0.6 ml each) were added to make the total volume 1.0 ml, and the cells were cultured at 37 ° C. for 4 days. As controls, HIV-1 non-infected cells and the compounds according to the present invention whose structures are shown in Chemical formulas 3 and 4 were used.

Cytopathic effect
measurement of the viable cell ratio in the virus-infected cells and non-infected cells after culturing for 4 days, using a tetrazolium compound (3- (4,5-dimethylthiazol-2-yl)-
It was determined by the absorbance of the formazan dye generated by the reduction of 2,5-diphenyl-tetrazolium bromide).

Further, the concentration of the compound that inhibits the growth of non-infected cells by 50% (cytotoxicity) and the virus-infected cells were 50%.
The concentration of the compound that survived (50% virus growth inhibitory concentration) was determined.

As a comparative example, the growth inhibitory activity was similarly measured for ZnCl ₂ and compounds 5, 6 and 7.
The obtained results are shown in Tables 1-6.

[0042]

[Chemical 5]

[0043]

[Chemical 6]

[0044]

[Chemical 7]

[0045]

[Table 1]

[0046]

[Table 2]

[0047]

[Table 3]

[0048]

[Table 4]

[0049]

[Table 5]

[0050]

[Table 6]

As shown in the results in Tables 1 to 6, the polyamine zinc complex having the structure shown in Chemical Formula 3 according to the present invention was added to HIV-1 at a concentration of 0 to 100.0 (μg / ml).
The viability of non-infected MT-4 cells was good, and when added at a concentration of 6.25 to 150.0 (μg / ml), the viability of HIV infected cells was good.

The cytotoxicity and 50% virus growth inhibitory concentration were 294 and 23 (μg / ml), respectively.

The polyamine zinc complex having the structure shown in Chemical formula 4 according to the present invention has a concentration of 12.5 to 800.0 (μg /
ml), the viability of HIV-uninfected MT-4 cells was good, and the concentration was 100.0-160.
When added at 0.0 (μg / ml), the survival rate of HIV infected cells was good.

The cytotoxicity and 50% virus growth inhibitory concentration were 2663 and 168 (μg / ml), respectively.

On the other hand, in each of the comparative examples, none of the compounds had a low cytotoxicity and showed a 50% virus growth inhibitory concentration.

Therefore, the polyamine zinc complex according to the present invention has low toxicity and high anti-HIV activity.

(Acute toxicity) An acute toxicity test by oral administration was carried out using ICR male mice (7-week life).
As a result of measuring the LD ₅₀ values of the compounds having the structures shown in Chemical formulas 3 and 4 according to the present invention, all were 1 g / kg or more, and high safety was confirmed.

INDUSTRIAL APPLICABILITY The polyamine zinc complex according to the present invention and the anti-acquired immunodeficiency syndrome virus agent containing the same have low toxicity and high activity against HIV (anti-human immunodeficiency virus) infected cells. Effective as an acquired immunodeficiency syndrome virus agent.

 ─────────────────────────────────────────────────── ─── Continuation of the front page (72) Eiichi Kimura 4-9-3 Misuzugaoka Higashi, Saiki-ku, Hiroshima City, Hiroshima Prefecture (72) Inventor Toru Koike 2-19-18 Ushidahigashi, Higashi-ku, Hiroshima City, Hiroshima Prefecture

Claims (3)

[Claims] 1. A polyamine zinc complex represented by the following structural formula 1. [Chemical 1] 2. An anti-acquired immunodeficiency syndrome virus agent comprising the polyamine zinc complex according to claim 1 as an active ingredient. 3. An anti-acquired immunodeficiency syndrome virus agent comprising a polyamine zinc complex represented by the following structural formula 2 as an active ingredient. [Chemical 2]