JPH02295997A - Thymine derivative and anti-hiv agent containing same - Google Patents

Abstract

NEW MATERIAL:A compound of formula I [R1 is lower alkyl or halogen; R2 and R3 are each tetradecanoyl(oxytetradecanoyl); R4 and R5 are each H or sulfonic acid group, but not H simultaneously]. EXAMPLE:1-(2-deoxy-2-tetradecanamide-3-O-tetradecanoyl-beta-D-glucopyr anosyl-4,6- disulfate)-5-methyl-1,2,3,4-tetrahydropyrimidine-2,4-dione. USE:Anti-HIV(human immunodeficiency virus) agents. Highly active as well as highly safe. PREPARATION:A thymine-glucosamine derivative of formula II as raw material is made to react in an inert solvent such as sulfonic acid (derivative) plus DMF at 45-50 deg.C for 20hr. It is suggested that the raw material compound be prepared from a thymine derivative of formula III as starting material.

Description

Detailed Description of the Invention [Field of Industrial Application] The present invention relates to a compound having antiviral activity, and more particularly to a thymine derivative having excellent anti-human immunodeficiency virus activity and an anti-human immune system containing the same. This invention relates to an anti-HIV agent (hereinafter abbreviated as an anti-HIV agent).

[Conventional technology and its problems]

Acquired Immunodeficiency Syndrome
eDeficiency
D S ] is human immunodeficiency virus Ct! uman
Immunodeficiency Virus.

Hereinafter abbreviated as HIV; Nature, 321.10
(I986)] is a serious immunodeficiency disease caused by infection with a very high mortality rate.
Countermeasures against HIV infection and AIDS have become a major social and financial challenge.

Azidothymidine (AZT), which has a reverse transcriptase inhibitory effect, is known as an anti-HIV agent that is currently recognized to be clinically effective, but its clinical efficacy is still insufficient, and This poses a problem in that side effects such as bone marrow (blood-forming tissue) disorders and neurological symptoms such as headaches and convulsions are severe. In particular, with HIV, the gene becomes a provirus and burrows into the chromosomes of infected cells, creating a condition similar to a genetic disease, which inevitably requires long-term administration of the drug, and AZT has such side effects. This is a major obstacle in using it as an anti-HIV agent.

Furthermore, by the time an HIV-infected person develops AIDS,
It usually has an extremely long clinical incubation period, which makes it extremely difficult to take measures to prevent infection.

Under these circumstances, the development of new pharmaceutical preparations that are effective against HIV infection and AIDS is eagerly awaited.

[Means to solve the problem]

Therefore, as a result of extensive research to solve the above problems, the present inventors discovered that a specific thymine-glucosamine derivative has a strong anti-HIV effect and is highly safe, and completed the present invention.

That is, the present invention relates to the following general formula (I) [wherein RI represents a halogen atom or a lower alkyl group,
R3 and R3 may be the same or different and represent a tetradecanoyl group or a tetradecanoyloxytetradecanoyl group, and R1 and R1 may be the same or different and represent a hydrogen atom or a sulfonic acid group, but R1 and R1 The present invention provides a thymine derivative represented by the following formula: and an anti-HIV agent containing the same as an active ingredient.

In the above general formula (I), the lower alkyl group represented by R1 includes an alkyl group having 1 to 5 carbon atoms, particularly a methyl group, an ethyl group, an n-propyl group, an isopropyl group, an n-butyl group, or An n-pentyl group is preferred. Further, examples of the halogen atom include a fluorine atom, a chlorine atom, a bromine atom, and an iodine atom.

Examples of the tetradecanoyloxytetradecanoyl group represented by R7 and R3 include 3-tetradecanoyloxytetradecanoyl group.

Further, this 3-tetradecanoyloxytetradecanoyl group has optical isomers, and includes all of the (R) form, (L) form, and racemic form.

Compound (I) of the present invention is produced, for example, according to the following reaction formula.

(n) (I) [In the formula, R1-R2 have the same meanings as above] That is, the thymine derivative (I) of the present invention can be obtained by reacting the thymine-glucosamine derivative (II) with a sulfonic acid or a derivative thereof. can be manufactured.

Sulfonic acid derivatives include sulfonic acid-tertiary amine complexes such as sulfonic acid-triethylamine complexes,
Examples include pyridine salts of sulfonic acids.

The reaction is carried out in an inert solvent such as dimethylformamide between 45 and
This is carried out by performing a patent reaction at 50°C for 20 hours.

The thymine-glucosamine derivative (n) as a raw material can be produced, for example, according to the following reaction formula.

Below is the blank space NHL; ] Each of the above reaction steps will be explained below.

(I) Compound (IV) is obtained by silylating thymine derivative (III).

This reaction is carried out by stirring the thymine derivative (III) and the silylating agent in a suitable solvent at -5 to 50°C. Examples of solvents used include dry benzene,
Examples include chloroform, methyl chloride, dioxane, and tetrahydrofuran. As a silylating agent,
Tri-lower alkylsilyl halides such as trimethylsilyl chloride, bistri-lower alkylsilylacetimides,
CIIC11.

Examples include C1130CO3i (CI+3) 2 .

(2) A thymine-glucosamine conjugate (VI) is obtained by reacting a silylated thymine derivative (IV) with a glucosamine derivative (V).

This reaction is carried out by reacting the silylated thymine derivative (IV) and the glucosamine derivative in a suitable solvent in the presence of a catalyst. Dichloroethane, chloroform, methylene chloride, etc. are used as the reaction solvent, and tin tetrachloride etc. are used as the catalyst. In addition, the reaction is
For example, it is preferable to carry out in the presence of a desiccant such as powdered Mohecular Leaves 48.

The glucosamine derivative used in this step can generally be synthesized using commercially available glucosamine as a starting material (although commercially available products may be used). Details of the synthesis method will be disclosed in Examples below.

(3) Compound (■) is obtained by hydrolyzing the thymine-glucosamine conjugate (VI).

This reaction is a hydrolysis reaction for removing the ester-bonded γ-cetyl group, and can be carried out by a method known per se. That is, it is carried out in the presence of a base such as sodium alkoxide, alkali hydroxide, or alkali carbonate.

(4) Compound (■) is obtained by acetalizing compound (■).

This reaction is carried out by reacting the compound (■) with an acetalizing agent such as acetone in the presence of a catalyst. Examples of acetalizing agents include para-toluenesulfonic acid, acetone, 2,2-dimethoxypropane, benzaldehyde, formaldehyde, acetaldehyde, and cyclohexane; examples of catalysts include hydrogen chloride, concentrated sulfuric acid, phosphorus pentoxide, and cation exchange resins. Examples include H" type zinc chloride and copper sulfate (n).

(5) Compound (IX) is obtained by hydrolyzing compound (■).

This reaction can be carried out by conventional hydrolysis methods such as acid-catalyzed hydrolysis, base-catalyzed hydrolysis, ammonolysis, transesterification, etc., but base-catalyzed hydrolysis is preferred. As the base catalyst, alkali hydroxide, alkali carbonate, sodium methoxide, etc. are used.

(6) Compound (X) can be obtained by reacting compound (IX) with tetradecanoic acid, tetrazocalyloxytetradecanoic acid, or a reactive derivative thereof.

When using tetradecanoic acid or tetrazocallyoxytetradecanoic acid directly in this reaction, it is preferable to use a dehydration condensation agent such as dicyclohexylcarbodiimide (DCC) or carbonylimidazole (DI3). Examples of the chemical derivatives include acid halides and acid anhydrides.The reaction is carried out using dimethylaminopyridine, 2.4.6-trimethylpyridine, triethylamine, etc. in an inert solvent such as dimethylformamide, methylene chloride, acetonitrile, and tetrahydrofuran. The reaction is preferably carried out in the presence of a base.

As for reaction conditions, it is preferable to react under water cooling for about 1 hour and then react at room temperature for about 200 hours.

(7) The raw material compound (XI) is obtained by deacetalizing compound (X).

This reaction is carried out by heating compound (X) in the presence of a suitable acid. Examples of acids that can be used include acetic acid, hydrochloric acid, and dilute sulfuric acid.

The heating temperature is preferably reflux temperature.

The thus obtained thymine derivative (I) has excellent anti-HI
It has V action and low cytotoxicity, so it is highly safe.
It is useful as an anti-HIV agent.

The anti-HIV agent of the present invention contains the above-mentioned thymine derivative (I) as an essential ingredient, and is usually prepared by blending the pharmacologically effective thymine derivative with a suitable pharmaceutical carrier.

As a pharmaceutical carrier, any excipient or diluent such as fillers, extenders, humectants, disintegrants, surfactants, etc. that are commonly used to prepare pharmaceuticals according to the usage form can be used. . The form of the pharmaceutical composition is not particularly limited as long as it effectively contains the above-mentioned active ingredients, and examples include solids such as tablets, powders, granules, and gunpowders, ordinary liquids, suspensions, and emulsions. It can be a liquid agent such as. It can also be made into a dry combustion product that can be made into a liquid by adding a suitable carrier before use. Various conventional additives such as solubilizing agents, buffering agents, soothing agents, preservatives, coloring agents, etc. may be added to the above pharmaceutical composition as necessary, and other pharmaceuticals may also be combined. You can also do that.

The anti-HIV agent of the present invention is administered by an appropriate administration route depending on the form of the pharmaceutical composition. The method of administration is also not particularly limited, and can be administered internally, externally, or by injection. Injectables are
For example, it can be administered intravenously, intramuscularly, subcutaneously, intradermally, intraperitoneally, etc., and external preparations include suppositories and the like.

The dosage of the anti-HIV agent of the present invention is determined appropriately depending on the formulation form, administration method, purpose of use, and the age, weight, medical condition, etc. of the patient to whom it is applied, and although it is not fixed, it is generally contained in the formulation. The amount of active ingredient administered per adult is approximately 0.1 g to 10 g for oral administration, and 0.1 g to 10 g for parenteral administration.
The amount is preferably about 1 g to 5 g, and the amount of active ingredient (2) in the preparation is appropriately set according to this dosage. Note that the administration can be divided into several times a day if necessary.

〔Effect of the invention〕

The compound (I) of the present invention has excellent anti-HIV action and low cytotoxicity, and is therefore useful as an anti-HIV agent. Therefore, the anti-HIV agent of the present invention containing this as an active ingredient is
It is also suitable for long-term administration and is extremely useful for preventing HIV infection and the onset and treatment of Al5D and its related syndrome ARC.

〔Example〕

The present invention will be further explained by referring to Reference Examples and Examples below.

Reference Example 1 Production of 1,3.4.6-tetra-0-acetyl-2-chloroacetamido-2-deoxy-β-D-glucobylanose: Stirred in an aqueous solution of IJI under water cooling. D-(+)-glucosamine hydrochloride 215.1g
(I,0011Iol) was gradually added and then p-
123.4 g (0.91 mo) of anisaldehyde was added dropwise, and after the dropwise addition, the mixture was stirred at room temperature for 1 hour. The formed crystals were filtered with suction and washed with cold water and then with ethanol:ether (I: 1). After drying, a white powder was obtained with a yield of 239.0 g and 81%.

The physical properties of the compound obtained in this step are shown below.

Melting point 177-178℃ (I) Bis(trimethylsilyl)thymine (IV-1
) Production: Thymine (manufactured by Wako Tsumugi Co., Ltd.) 6.00 g (0,048
mol) and trimethylchlorosilane (manufactured by Shin-Etsu Chemical Co., Ltd.) 12.4 g (0.115a+ol) in dry benzene 100mj! Triethylamine 11 is dissolved in this and triethylamine 11
．． 6g (0,115mat) was gradually added dropwise. The reaction solution was suction filtered using Celite (manufactured by Wako Tsumugi Pharmaceutical Co., Ltd.).
After concentrating the filtrate under reduced pressure, it was placed in a glass tube oven (5
Distillation under reduced pressure (initial distillation 75°C, 35 m
m11g; water distillation 150°C, 9-10 mmHg) L
, 10.9 g of the target compound (I'V-1) was obtained. Yield 8
2.8%.

The physical properties of the compound obtained in this step are shown below.

Boiling point: 98℃/2 +r+n+lIg melting point near 3-7
4°C H-NMR (CDC1!3) δppm: 0.36
(I8H,s, SiMeffx 2) Example 1 (2) 1-(3,4,6-)20-acetyl-
Production of 2-chloroacetamido-2-deoxy-β-D-glucopyranosyl)-5-methyl-1,2,3,4-tetrahydropyrimidine-2,4-dione (■-1): Obtained in (I) Compound (■-1) 2.06 g (
7,5X l O-' mol), 2.12 g of 1,3,4.6-tetra-0-acetyl-2-chloroacetamido-2-deoxy-β~D-glucobylanose obtained in Reference Example 1 (5,0 x 10-' mol) and 1.50 g of powdered molecular sieves 48 at 1.2-
It was added to dichloroethane 401111 and stirred at room temperature for 1 hour. The mixture was then cooled with ice under an argon atmosphere, and 1.95 g (7,5 x 10-3111 ol) of tin tetrachloride was gradually added dropwise, followed by stirring for 1 hour under water cooling and further stirring at room temperature for 15 hours.

After confirming the completion of the reaction by thin layer chromatography (developing solvent: chloroform:acetone-10:2), saturated N
a1lCL aqueous solution was added, stirred for 1 hour, and celite (
The mixture was filtered by suction using a filter (manufactured by Wako Tsumugi Pharmaceutical Co., Ltd.).

After washing the filtrate with saturated NatlCO=, the organic layer was concentrated under reduced pressure, and the resulting residue was subjected to silica gel column chromatography (100 g of silica gel, developing solvent: chloroform:acetone=10:2 → chloroform:methanol).
10:l → 5:l → 3:l) to obtain the target compound (V
l-1) 1.56 g was obtained.

Yield 63.6%.

The physical properties of the compound obtained in this step are shown below.

Melting point: 132-135℃ Elemental analysis (C, l + 2.N, [1, O (as :i)
:HN Calculated value (%) 46.59 4.94 8.58
Actual value (%) 46.74 5.10 7.62
1Rν,,, -': 3320. 1745.
1700, 1670I11'H-NMR (CDC13) δppm: 2.11
(9N, s, ^cOX3)3.95 (2
H, s, CI Cl1=C-)5.55 (I)
1. d, J=9.0 1-1z.

Mouth NH[:CH,Cj! ) ”C-Ni4R (CDC1! 3) δppm: 42
．． 3 (CI CH,C) 80.8 (Anomeric carbon) 112.4 (Cs) 135.1 (C,) 151.5 (C2) 163.5 (C4) 167.1, 169.4, 170.6 .

170.8 (C=0) (3) 1-(2-chloroacetamido-2-deoxy-
β-D-glucopyranosyl)-5-methyl-1,2,3
, 4-tetrahydropyrimidine-2,4-dione (■-
Production of 1): 2.70 g of the compound (Vl-1) obtained in (2) (
5,5X 10-3 mol) in dry smoked methanol 150m
j! 0.1 M under argon atmosphere and water cooling.
55ml of methanol solution of sodium methoxide! was added dropwise to a water-cooled tube, stirred for 1 hour, and further stirred at room temperature for 20 hours.

After confirming the completion of the reaction by thin layer chromatography (developing solvent: chloroform:methanol = 3:1), a weakly acidic ion exchange resin ICR-50 (manufactured by Organo) was used.3.
00 g was added and stirred for 1 hour. After removing the ion exchange resin by filtration and concentrating the filtrate under reduced pressure, the resulting residue was subjected to silica gel column chromatography (silica gel LOOG, developing solvent: chloroform:methanol = 10:1 → 5:1 →
3:1) to obtain 52 g of the target compound (■-1). Yield 76.0%.

The physical properties of the compound obtained in this step are shown below.

Melting point: 143-147°C 'H-NMR (CDCj! a) δppm: 3.9
8 (211,s, CjICIIiC)5.70
(ltl, d, JJ, 5 Hz.

C-NMR (CDC1s) δppm: 43.1
(CA' CH, -) 82.7 (Anomeric carbon) 111.6 (C,) 138.6 (Cs ) 152.6 (C,) 166.2 (C,) 169.9 (C=0) ( 4) 1-(2-chloroacetamido-2-deoxy-
Production of 4,6-itupropylidene-β-D-glucopyranosyl)-5-methyl-tetrahydropyrimidine-2,4-dione (■-1) = Compound obtained in (3) (■-
1 > 1.52 g (4,188IO-' and 2.2
-dimethoxypropane 3.27 g (3,14X 10
-2 mol) in DMF30n+1, p-)luenesulfonic acid 0.33 g (I,94xio-"■
ol) was added and stirred at room temperature for 20 hours.

Can you complete the reaction using thin layer chromatography (developing solvent: chloroform:methanol = 5:1)? ? [After that, I
2.50 g of RA 45 (manufactured by Organo) was added and stirred for 1 hour. After the reaction solution was filtered naturally and the filtrate was concentrated under reduced pressure, the resulting residue was subjected to silica gel column chromatography (30 g of silica gel, developing solvent;

Chloroform:methanol = 10:1 → 5:1 → 3:1
) to obtain 118 g of the target compound (■-1). Yield 77.5%.

The physical properties of the compound obtained in this step are shown below.

Melting point: 219-221°C Elemental analysis (C+-1liJsLcj?H, (as l): HN Calculated value (%) 45,56 5.73 9.96
Actual value (%) 45,93 5.39 10.13
'H-NMR (CDC1's) δ ppm
:1.46. 1.57 (each 3H,s.

3.97 (2H,s, CjICHzC)5.7
6 (I)1. d, J=9.9 fiz.

CIC1, CNH) (5) 1-(2-amino-2-deoxy-4,6-imbropylidene-β-D-glucopyranosyl)-5-methyl-1,2,3,4-tetrahydropyrimidine-2 ,4
-Manufacture of dione (IX-1): Compound (■-1) obtained in (4) 1.18g (2,92X 10-3ma
t), thiourea 0.44 g (5,84x10-'
mol), diisobrobylethyramine 0.76g
(5,84X 10-3 mol) and powdered %L/
2.0 g of Curar Sieves 4A (manufactured by Wako Tsumugi Yakuhin Co., Ltd.) was added to 40 ml of tetrahydrofuran, and under an argon atmosphere,
The mixture was heated under reflux at room temperature for 3 hours.

After confirming the completion of the reaction by thin layer chromatography (developing solvent: chloroform:methanol = 5:1), suction filtration was performed using Celite (manufactured by Wako Tsumugi Pharmaceutical Co., Ltd.), and the filtrate was condensed under reduced pressure to obtain the The residue was purified by silica gel column chromatography (silica gel 50 g, developing solution AK; chloroform:methanol=10:I→IO: 2) to obtain 07 g of the target compound (■-1). Yield 8
5.2%.

The physical properties of the compound obtained in this step are shown below.

Melting point: 98-102°C IR C-x-': 3340. 1661
．． 858m'H-NMR (CDCj! 3) δppm: 1.4
7. 1.59 (each 311.s.

"C-NMR (COCA's) δppm: 1.9
1.28.9 (acetonide) 81.8 (anomeric carbon) 112.0 (Cs ) 136.5 (C,) 152, O (C2) 164.7. ][i4.5 (C,)170.7
, 170.0 (N C-0) ((3) 1-(2-
Deoxy-4,6-itubropyrimidine-2-tetradecanamide-3-〇-tetradimethyl-β-D-glucopyranosyl)5-methyl-1,2,3,4-tetra-hydropyrimidine-2,4- Production of dione (X-1): (
5) Compound (IX-1) obtained in 0.15 g (4
, 5X I O-' mol), tetradecanoic acid 0.
31 g (I,35X 10-' mol) and dimethylaminopyridine 0.05 g (I,35X 1
0-' mol) in dimethylformamide 5. Omj!
and 2.0mj of methylene chloride! Dissolved in a mixed solvent of
The mixture was stirred for 1 hour and further stirred at room temperature for 15 hours.

After confirming the completion of the reaction by thin layer chromatography (developing solvent: chloroform:acetone=10:2), the reaction solution was concentrated under reduced pressure, 2 ml of ethyl acetate was added, and suction filtration was performed using Celite. The filtrate was again concentrated under reduced pressure, and the resulting residue was subjected to silica gel column chromatography (20 g of silica gel, 1 developing solvent; chloroform: acetone = 1
0: Purified by 2) to obtain target compound (X-1) 0.12
I got g.

Yield 35.7%.

The physical properties of the compound obtained in this step are shown below.

'H-NMR (CD (J) δppm: 0.88 (
6)1. t, J = 5.6 Hz.

(Ctla) I 2 CH3X 2 ) 1, 26
(4811,brs, -(CHa) +2CII3X
2) 1.49. 1.53 (each 311.
s.

['112-X2) (7) 1-(2-deoxy-2-tetradecanamide-3
-0-tetradecanoyl-β-D-glucopyranosyl)
Production of -5-methyl-1,2,3,4-tetrahydrobilidine-2,4-dione (II-1): (6) 0.10 g of compound (X-1) (
I,3X 10-' mol) was dissolved in 1 mml of a 9:1 mixture of acetic acid and water, and the mixture was refluxed at 90 to 95°C for 1 hour.

Purification was performed by thin layer chromatography (20 g of silica gel, 1 developing solvent; chloroform:methanol=10:1) to obtain 0.55 g of the target compound (II-1).

Yield 54.3%.

(8) 1-(2-deoxy-2-tetradecanamide-3
-〇-tetradecanoyl β-D -/'rucobyranosyl-4,6-disulfe 1-) -3-l f-1,2,3.4-tetrahydropyrimidine-2,4-dione (I-1) Production: Compound obtained in (7) (n-
1) 30 mg (4,24X 10-' mat) and ammonium trimethylsulfonate 40B (2,87X 10-'
-' mol) and dimethylformamide 1.5 m
J! and stirred at 50°C for 24 hours.

After confirming the completion of the reaction by thin layer chromatography (developing solvent: chloroform:methanol=10:1), it was concentrated under reduced pressure, and the residue was dissolved in methylene chloride 1. 14 mg of trifluoroacetic acid (I, 26X
10-'mol> was added and stirred for 1 hour. The reaction solution was then concentrated under reduced pressure, and the resulting residue was purified twice by gel chromatography (5 ephadex LH2020g1 developing solvent; chloroform:methanol=1:I) to obtain 16 mg of the target compound (I-1). Yield 36.0%
.

The physical properties of the compound obtained in this step are shown below.

IRλ,,, ctx-': 1710 (,;
C=0), 1650(acid>, t25o
c;s=u.

800 (C-0-S-N Example 2 (I) l-(2-deoxy-4,6-itubropylidene-2- ((R)-3-tetra-decanoyloxytetradecanamide)- 3-0-((R)3-tetradecanoyloxytetradecanoyl)-β-D-glucopyranosyl]-5-methyl-1,2,3,4-tetrahydropyrimidine-2,4-dione (X-2 ) Production of Compound (IX-1) obtained in Example 1 (5) 0.
17g (0.5xl Q-3mol), (R)-3-
f Tradecanoyloxytetradecane! ! 0.55g
(I,2x10-" mat) and dimethylaminopyridine 0.061 g (0,5X 10"' mol
) was dissolved in 5 tnl of acetonitrile, and 0.25 g of dicyclocarbodiimide (
I,2×10-' mol) was added. Then, under water cooling
The mixture was further stirred at room temperature for 15 hours. The reaction solution was concentrated under reduced pressure, 10 ml of ethyl acetate was added to the residue, and suction filtration was performed using Celite. The filtrate was again concentrated under reduced pressure for 11 minutes, and the resulting residue was subjected to silica gel column chromatography (20 g of silica gel, developing solvent: chloroform diacetone = 2
0:1) to obtain 50 mg of the target compound (X-2). Yield 8.3%.

The physical properties of the compound obtained in this step are shown below.

'H-NMR (CDCji!s) δppm: 1.2
6 (88)1. brs, (CH2)44-)-
C-CH2-) (2) 1-[2-deoxy-2-((R)-3-tetradecanoyloxytetradecanamide)3-0- ((
R)-3-tetradecanoyloxytetradecanoyl)
-β-D-glucopyranosyl]-5-methyl-1,2,
3,4-tetrahydropyrimidine-2,4-dione (I
Production of I-2): Compound (X-2) obtained in (I) 0.20 g (
I,66X 10-' mol) was dissolved in a mixture of acetic acid:water=9:I and refluxed at 90 to 95°C for 1 hour.

The product was purified by thin layer chromatography (15 g of silica gel, 1 developing solvent; chloroform:methanol=20:1) to obtain 75 mg of the target compound (II-2). Yield 38.7
%.

The physical properties of the compound obtained in this step are shown below.

'II-NMR (CDCj!,) δppm: 0.8
9 (I211,t, J=4.8411z.

(CI+ 2) -, CI+ 3) 1.23 (8Jl11.brs, (I:IIz
L-)1l -C-C1,-) (3)! -(2-deobushi2- ((R)-3-tetradecanoyloxytetradecanamide)3-0- ((
R)-3-tetrazo°kayloxytetradecanoyl)-β-D-glucopyranosyl-4,6-disulfate]-5-methyl-1,2,3,4-tetrahydropyrimidine-2,4-dione Production of (I-2>: Compound (II-2) obtained in (2) 75 mg (6,
5xl Q-' 0101) ammonium trimethylsulfonate 100B (7,2810-' 111o
t) in 1.5 mf of dimethylformamide,
The mixture was stirred at 0°C for 20 hours.

After confirming the completion of the reaction by thin layer chromatography (developing solvent: chloroform:methanol-2=l), it was concentrated under reduced pressure, and the residue was further dissolved in 1 Oml of methylene chloride, and 30 mg of trifluoroacetic acid (2,6X 10-'
mol) of methidichloride: /1 mj? A solution dissolved in was added and stirred for 1 hour. Next, the reaction solution was concentrated under reduced pressure, and the resulting residue was subjected to gel chromatography.
x LII20 20 g, developing solvent; special grade chloroform: dry smoke methanol = l: 1) and purified twice to obtain 57 mg of the target compound (I-2).

Yield 66.8%.

The physical properties of the compound obtained in this step are shown below.

IRλm,,lccm-': 1710 (,"
C=0), 1660 (acid), 1213
0 (n5=O).

810 (C-0-3) Test Example 1 Examination of anti-HIV effect: In this test example, the anti-HIV activity of dextran sulfate (Kowa ■) and the compound of the present invention (I-2) was evaluated using MT-4 cells. Judgment system (Nagamo, T.

and tloshino H,; Jpn, J.
Cancer Res, 79.9-11゜1988)
I looked it up.

That is, the test substance was added to 1X10' MT-4 cells, infected with HIV, and reacted for 4 days at 37°C under 5% CO2. Then, a smear of MT-4 cells was prepared, and Hf
The degree of establishment of V infection was determined by indirect fluorescent antibody method. The results are shown in Table 1 as a comparison with the control in which no test substance was added.

The compound of the present invention (I-2) has a very small molecular weight compared to dextran sulfate, which is a sulfated polysaccharide.The compound of the present invention (I-2) has a clear anti-tl IV effect, Even at a concentration of 100 μg/mi, no growth inhibition or toxicity to MT-4 cells was observed.

Test Example Examination of anti-HIV effect: In this example, the anti-HIV activity of azidothymidine (AZT) and the compound of the present invention (I-2) was investigated by culturing a mixture of HIV-infected cells and HIV-uninfected cells, and examining the formed compound. Cell body (S
Syncytium As5ay method was used to measure the number of syncytium cells.

That is, MoI t-4 cells (J.

Minowada et al, J, Natl C
ancer In5t (J, N, C0I).

49, 891-895.1972) and added each test substance to Molt-4 cells 1. which had been infected with HIV in advance.
Add 5 x 10' pieces and add this to 37 cm under 5% CO2.
After incubating overnight at °C, formaldehyde was added to a final concentration of 5%, fixed, and the number of syncytia formed (those whose diameter had increased five times or more) was counted. So.

The results are shown in Table 2.

Below, margin AZT already showed cytotoxicity at 10 μg/rnl, and syncytial formation was often observed even at about 100 μg/ml. On the other hand, the compound of the present invention (I-2>) showed no cytotoxicity even at a concentration of 100.ug/ml.Moreover, the formation of syncytia was effectively inhibited at low concentrations, demonstrating its efficacy and safety. You can see that it has both genders.

that's all Applicant: Japan Antibody Research Institute, Inc.

Claims (1)

[Claims] 1. The following general formula (I) ▲There are mathematical formulas, chemical formulas, tables, etc.▼ (I) [In the formula, R_1 is a halogen atom or a lower alkyl group, and R_2 and R_3 are the same or different. R_4 and R_5 may be the same or different and represent a hydrogen atom or a sulfonic acid group, but R_4
and R_5 do not become hydrogen atoms at the same time.] A thymine derivative represented by: 2. An anti-HIV agent containing the thymine derivative according to claim 1 as an active ingredient.