JPH04108788A - Antiviral quaternary ammonium salt and its production - Google Patents

Abstract

NEW MATERIAL:A compound expressed by formula I (X is halogen; Y is H or amino). EXAMPLE:(9-beta-D-2', 3'-dideoxyribofuranosylpurin-6-yl)-trimethylammonium chloride. USE:Useful as a treating agent or a preventive agent of acquired immunodeficiency syndrome (AIDS). PREPARATION:A purine compound expressed by formula II (e.g. 6- chloropurine-9-beta-D-2', 3'-dideoxyribofuranoside) is treated with trimethylamine in an organic solvent (e.g. benzene).

Description

DETAILED DESCRIPTION OF THE INVENTION [Industrial Application Field] The object of the present invention is to provide a novel antiviral ammonium salt represented by the general formula [I] and a method for producing the same.

Mex"X- (In the formula, X is a halogen atom, and Y is either a hydrogen atom or an amino group) -N"Me3・X-) 2', 3'
- There are no known reports regarding dideoxypurine nucleosides and their synthesis.

Furthermore, there are no known reports regarding antiviral effects.

[Problem to be solved by the invention]

Viral diseases such as hepatitis B, herpes, and adult T-cell leukemia are a major problem around the world.

Among these, AIDS caused by infection with HIV has a high mortality rate and is the most alarming for humankind.

There is still no known complete cure for AIDS, but AZ
Some drugs, such as T, are known to be effective in delaying the progression of the disease or preventing its onset.

A Z T (Proc, NN1. Acgd, Sc
i, USA 82.71195 (191151) is the only drug currently approved for use, and is an excellent drug that is also effective in improving AIDS dementia, but the drawback is that it is not strong against bone marrow cells. It is known to be toxic.

d d I (Proc, N!tl^), which is currently in clinical practice, was developed as a drug with less bone marrow toxicity than AZT.
c1d, sci, USA 83. 1911 (19
116)). However, ddI is less stable against acids than AZT and has low fat solubility, so it cannot be expected to be effective in treating AIDS dementia.

Furthermore, a common drawback of AZT and ddl is poor solubility in water.

The degree of water solubility is a factor related to the amount and concentration of the injection solution during intravenous administration. Also, in the case of oral administration, it is deeply involved in setting the concentration of the drug solution.

Since AZT and ddI exhibit only low solubility in water, they are inconvenient from a practical standpoint.

For example, in the case of intravenous injection, in order to raise the concentration in the blood above the effective concentration, there are problems such as the dilution of the injection solution and the need for a considerable amount of liquid, and the long time required for the infusion. It was seen.

An object of the present invention is to provide a novel antiviral agent that can overcome the various drawbacks of existing drugs, and a method for synthesizing the same.

[Means for Solving the Problems] The present invention provides a novel nucleic acid represented by the general formula [I] (wherein X is a halogen atom and Y is either a hydrogen atom or an amino group) It is based on the discovery that derivatives have antiviral activity.

The present invention is a novel antiviral agent characterized by containing as an active ingredient at least one type of antiviral quaternary ammonium salt represented by the above general formula [I], and a method for producing the same. It is related to.

In the present invention, the purine derivative represented by the general formula [Ir] can be easily obtained using a base exchange reaction using a microorganism (Japanese Patent Application No. 1-46183).

In addition, if immobilized microorganisms are used, it can be obtained industrially at low cost (Japanese Patent Application No. 181885/1999).

[For production]

The antiviral action of the novel antiviral quaternary ammonium salt of the present invention will be explained below.

The antiviral effect of the antiviral quaternary ammonium salt of the present invention is mainly due to the competitive inhibitory effect of its structure (2',3'-dideoxypurine nucleoside skeleton) and the D
This is due to the NA chain termination effect.

In other words, the antiviral quaternary ammonium salt of the present invention has a structure that is very similar to DNA-constituting nucleic acids (2'-deoxyadenosine, 2'-deoxyguanosine, etc.), so it has a high affinity for virus enzymes. very good. Therefore, it competitively inhibits the virus by slowing down its own DNA synthesis.

Furthermore, the antiviral ammonium salt of the present invention is a 3'-
Because it has a structure in which there is no hydroxyl group at the position, -
Once incorporated into the viral DNA, the DNA chain ends and the virus dies. That is,
The 2'3'-dideoxypurine nucleosides of the present invention act as DNA chain terminators and exhibit antiviral activity.

AZT, which is currently used as a treatment for AIDS,
Clinically available drugs such as ddI also have a similar effect on competitive inhibition and DN.
The 2',3'-dideoxypurine nucleosides of the present invention exhibit antiviral activity through A-chain termination, but these existing drugs (
AZT.

ddl, etc.).

That is, the 2',3'-dideoxypurine nucleosides of the present invention have a special quaternary amine salt structure called a trimethylamine salt at the 6-position of the purine base, and therefore have very good solubility in water. Easily dissolved even at a high concentration of 5g/1ml. This value is so large that it cannot be compared with the saturation concentration of AZT or ddI (approximately 2 to 10 .mu./ml). Due to this high solubility in water, it is possible to significantly reduce the amount of drug for intravenous injection.

Furthermore, since quaternary ammonium salts are electrolytes, they are not only easily soluble in water, but also have good permeability to cell surfaces. This helps drugs that enter the bloodstream reach cells.

In addition, when taken orally, it can aid absorption through the stomach wall and intestinal mucosa.

The trimethylammonium salt located at the 6-position of the purine base attracts electrons due to its quaternary ammonium salt effect, reducing the electron density of the purine base itself. Therefore, the electron density near the glycosidic bond of the antiviral quaternary ammonium salt of the present invention is also reduced. As a result, (d
stability against acids (compared to dI, etc.) can be increased.

Several natural compounds with trimethiamine salt structures are already known, such as the neurotransmitter acetylcholine and the mushroom toxin muscarine. What these compounds have in common is that, despite their relatively high water solubility, they are able to reach the nervous system and brain.

The antiviral quaternary ammonium salt of the present invention also behaves in a similar manner in vivo and acts directly on the nervous system. Therefore, it can directly act on the AIDS virus hidden in the brain and nerve cells, making it very effective in improving dementia.

In addition, methyl group donors such as methionine and betaine are
There are reports that it improves acquired abnormalities caused by HIV.
he Lancet 335 (19901).

The novel antiviral quaternary ammonium salts of the present invention are also methyl group donors and are therefore particularly effective in pediatric AIDS patients infected with HIV.

As described above, the antiviral quaternary amine salt of the present invention has a special structure of trimethylamine salt in its molecule, so it exhibits excellent water solubility and has good reachability and absorption into cells. It also has properties such as good stability against acids. It is also a new antiviral agent that is expected to improve dementia caused by AIDS.

The novel antiviral quaternary ammonium salt of the present invention is particularly effective against HIV, the virus that causes AIDS, but is also effective against other retroviruses (such as R3V, the sarcoma virus of L. It also shows excellent antiviral effects against the leukemia virus HTL~r-1). It also shows antiviral effects against viruses that have reverse transcriptase, such as hepatitis B virus. In addition, herpesviruses (for example, F (SV-1, H
3V-2, etc.) It also shows antiviral effects against viruses that do not have reverse transcriptase.

The 2',3'-dideoxypurine nucleoside having a halogen at the 6-position, which is used as a raw material in the present invention, can be easily obtained using a base exchange reaction using a microorganism (Japanese Patent Application No. 1-46183').

For example, when 6-chloroguanine and dideoxyuridine are used as raw materials, microorganisms (such as Eseheri
When a base exchange reaction is performed using chia eoli),
The reaction product 2',3'-dideoxy-6chloroguanosine is obtained in good yield. Furthermore, it can be obtained even more easily by using immobilized microorganisms (Japanese Patent Application No. 181885/1999).

2 having a halogen at the 6th position synthesized as above
The antiviral quaternary amine salt of the present invention can be easily synthesized using a ',3'-dideoxypurine nucleoside, a suitable organic solvent (eg, benzene), and trimethylamine.

For example, 6-chloro-2',3'-dideoxypurine ribofuranoside is dissolved in hot benzene to make a homogeneous solution, and after returning to room temperature, trimethylamine is added and 1
When left in the sun, chlorine at the 6-position reacts with trimethylamine, and the target product (9-β-D-2', 3'-dideoxyribofuranosylpurin-6-yl)-trimethylammonium chloride is produced in a benzene solution. It precipitates out from

When this is collected and dried, it is obtained as a white powder.

As described above, according to the synthesis method of the present invention, the antiviral agent related to the present invention can be synthesized by a simple method from readily available raw materials.

〔Example〕

The present invention will be explained in more detail below with reference to Examples.

However, the following examples are provided for illustrative purposes only and are not intended to limit the scope of the present invention in any way.

(1) Production Example Production Example 1 6-chloropurine-9-β-D-2',3'-dideoxyribofuranoside (Patent Application No. 1-46183) j, D
g (3,93rpvoI) in 15 ml of anhydrous benzene and 1 ml of dimethylformamide (hereinafter abbreviated as DMF).
Completely dissolve in a mixed solvent consisting of: After confirming that there is no undissolved matter and the solution is in a homogeneous state, add trimethylamine] Oml'& at room temperature.

Approximately 30 minutes after the completion of the dropwise addition, the inside of the reaction system becomes cloudy. The mixture is left as it is at room temperature for 12 hours, and the precipitated solids are filtered off, washed with benzene, and then dried under reduced pressure.

The obtained white solid content was dissolved in a mixed solvent of benzene and DMF (benzene/DMF=10/1).
Recrystallization from 1. IOg (3,22mn
ol) was obtained. Yield 82%.

White microcrystals Melting point 136°C NMR spectrum (1) 20): δ1.75-2.
6 (4H, m, C2'and C3'-H)3
．． 4-3.85 (2H, m, C5'-H) 3.75
(9H,s, NC-H)3.8~4.3 (IH
, m, C4'-H)51θ (18,t, C5'-
0)()6.20 (IH, t, C1'-H)
7JO (2H, s, NH2) 8.70 (IH, s, CB -H) MS spectrum (FAB-MS: m#) M-CI 293 (as C13H2102N 6+) Rf value (Silica Gel HF 254) 0
．． 0 [CH(, / 3 /MeOH=9/130.1
1 [MeOH/H20=9/1] Production Example 2 2-amino-6-chloropurine-9-β-D-2'.

3'~Dideoxyribofuranoside (Patent application No. 1-4618)
3) 1.0g (3,7111101) was added to 10ml of anhydrous benzene. Completely dissolve in a mixed solvent consisting of 5 ml of dimethylformamide (hereinafter abbreviated as DMF). After confirming that there is no undissolved matter and the solution is homogeneous, 10 ml of trimethylamine is added at room temperature. Approximately 30 minutes after the completion of the dropwise addition, the inside of the reaction system becomes cloudy. The mixture is left as it is at room temperature for 12 hours, and the precipitated solids are filtered off, washed with benzene, and then dried under reduced pressure.

The obtained white solid content was dissolved in a mixed solvent of benzene and DMF (
Recrystallized from benzene/DMF=3/1) to obtain (2-amino-9-β-D-2', 3'dideoxyribofuranosylpurin-6-yl)-trimethylammonium.
1. Chloride as white microcrystals. (12g (3,
I got 10Illio.

Yield 84%.

White microcrystals Melting point 147°C NMR spectrum (D2o); δ 1.8-2.8 (4H, m, C2'and C
3'-H) 3.5-3.7 (2H, m, C5'
-H) 3.75 (9H,s, NC-H)4.
0-4.4 (IH, m, C4'-H)525
(LH, t, C5'-0H)6.50 (I
H,t, cl'-H)9.10 (IH,s,
C8-H) 9.30 (IH, s, C2-H)
MS spectrum (FAB-MS; I11#) MC
I 278 (CI3H2002N, as 10) Rf value (Silica Gel HF 254) 0
．． 0 [CHCl3 /Me 0H=9/1]Q,I
[M e OH/H20 = 9/1 (2) Antiviral test Antiviral data of representative compounds according to the present invention are shown below. The compound number indicates the number of the above-mentioned example illustrating the preparation method in the Examples of the present invention.

Test Example 1 The antiviral effect of the compound represented by formula [I] of the present invention was tested using Rous sarcoma virus (RS V).

Primary culture cells (Cbicb Enb7o Fibrol
xs[) for about 30 minutes. Then, stepwise diluted samples were added, and after 4 to 7 days, R3V
The stage at which cell transformation due to infection was controlled was determined by microscopy. The results are shown in Table 1.

Anti-R3V active compound number of 2',3'-dideoxypurine ribofurinoside of the present invention Abbreviation r D, o (μM) CD50 (μM) As shown in Table 1, the compound of the present invention has excellent antiviral effects. It is clear that the compound of the present invention is effective as a therapeutic agent for viral diseases such as AIDS and hepatitis B because it has very good solubility in water.

(3) Solubility test The compounds synthesized in Production Examples 1 and 2 were tested for solubility in water as follows.

Four drugs, namely ■AZT ■ddl ■(9-β-D-2',3'-dideoxyribofuranosyl-6-yl)-trimethylammonium chloride■(2-aminy9-β-D-,2 ', 3'-dideoxyribofuranosyl-6-yl)-trimethylammonium chloride were taken at room temperature (50°C) to dissolve them.
) The amount of ultrapure water needed for each was measured using a microsyringe.

The results are as follows: 01mg15HμJ ■1■/100μl ■10■/〈5μl ■l0mg/<5μノ As shown above, compounds ■ and ■ have excellent hydrophilicity, which helps in drug formulation and improves absorption. It can be said to be a new type of antiviral agent, with excellent antiviral properties.

(4) Acid stability test pH 3,6 adjusted with hydrochloric acid and sodium hydroxide
A stability test was conducted using an aqueous solution of .

Reagent A (9-β-D-2', 3'-dideoxyribofuranosyl-6-yl)-trimethylammonium chloride Reagent B (2-amino-9-β-D-2', 3'-
dideoxyribofuranosyl-6-yl)-trimethylammonylam chloride A >5h >1 week >l
weekB >5b >], week
>l weekddl 15m1n
>1 da7 >l week As shown above, it can be seen that compounds A and B are sufficiently stable even at 9)+3 when compared to ddI.

〔Effect of the invention〕

As explained above, according to the present invention, a novel compound with excellent antiviral activity is provided. For example, it is extremely effective as a therapeutic agent for viral diseases such as AIDS, hepatitis B, and herpes.

Procedural amendment (Contents of the amendment October 23, 1990 1゜Indication of the incident 1990 Patent Application No. 223755 2, Title of the invention Antiviral quaternary ammonium salt and its manufacturing method 3,
Relationship with the case of the person making the amendment Patent applicant address: 4-5 Marunouchi-chome, Chiyoda-ku, Tokyo
Name (234) Sanyo Kokusaku Valve Co., Ltd. 4゜

Claims (9)

[Claims] (1) General formula [I] ▲There are mathematical formulas, chemical formulas, tables, etc.▼ Antiviral properties shown by [I] (In the formula, X is a halogen atom and Y is either a hydrogen atom or an amino group) Quaternary ammonium salt. (2) (9-β-D-2',3'-dideoxyribofuranosylpurin-6-yl)-trimethylammonium chloride (2-amino-9-β-D-2'-,3'-dideoxyribofuranosylpurin-6-yl) 2. A compound according to claim 1 selected from furanosylpurin-6-yl)-trimethylammonium chloride. (3) General formula [II] ▲ There are mathematical formulas, chemical formulas, tables, etc. ▼ [II] (In the formula, X is a halogen atom and Y is either a hydrogen atom or an amino group) The purine compound shown in A method for producing a nucleic acid derivative, which comprises producing an antiviral quaternary ammonium salt represented by the general formula [I] by treatment with trimethylamine in an organic solvent. ▲There are mathematical formulas, chemical formulas, tables, etc.▼ [I] (In the formula, X is a halogen atom, and Y is either a hydrogen atom or an amino group) (4) As the organic solvent used in the reaction according to claim 3, the purine compound represented by the general formula [II] can be completely dissolved, and the antiviral ammonium salt represented by the general formula [I] produced by the reaction is used. A method for producing a nucleic acid derivative, characterized by using a liquid that can be precipitated. (5) A method for producing a nucleic acid derivative, wherein the organic solvent used in the reaction according to claim 3 is benzene or dimethylformamide. (6) An antiviral agent containing at least one type of antiviral quaternary ammonium salt represented by the general formula [I] according to claim 1 as an active ingredient. (7) An antiretroviral agent containing at least one type of antiviral quaternary ammonium salt represented by the general formula [I] according to claim 1 as an active ingredient. (8) A therapeutic and prophylactic agent for acquired immunodeficiency syndrome (AIDS), which contains at least one type of antiviral quaternary ammonium salt represented by the general formula [I] according to claim 1 as an active ingredient. (9) Experimental medicines and experimental reagents used in genetic engineering, which contain at least one kind of antiviral quaternary ammonium salt represented by the general formula [I] according to claim 1 as an active ingredient.