KR100374326B1 - Human HIV Protease Inhibiting Compounds Having 6-Hydroxy-1,3-dioxin-4-one Ring and Process for Preparing the Same - Google Patents

Abstract

The present invention relates to a compound having a 6-hydroxy-1,3-dioxin-4-one ring that inhibits a protease, an essential enzyme for human immunodeficiency virus (HIV), and a method for preparing the same. Compounds having a 6-hydroxy-1,3-dioxin-4-one ring of the present invention are excellent in inhibiting HIV protease and antiviral activity, and have a markedly low cytotoxicity, resulting in AIDS or HIV infection. It may be usefully used in the development of therapeutic agents for the treatment or prophylaxis.

Description

Human HIV Protease Inhibiting Compounds Having 6-Hydroxy-1,3-dioxin-4-one Ring with 6-hydroxy-1,3-dioxin-4-one ring and Process for Preparing the Same}

The present invention relates to compounds having a 6-hydroxy-1,3-dioxin-4-one ring. More specifically, the present invention provides a compound having a 6-hydroxy-1,3-dioxin-4-one ring of general formula (I) below, which inhibits protease, an essential enzyme for human immunodeficiency virus (HIV), and It relates to a manufacturing method thereof.

HIV, which causes AIDS, is a retrovirus with genetic information in the form of RNA. Invading host cells, the virus requires several essential enzymes in the process of replicating new viruses, one of which is the protease. Proteases are enzymes essential for the breakdown of complex proteins formed during viral replication into structural proteins and viral enzymes (see Kohl, NE et al., Proc. Natl. Acad). Sci., USA, 85: 4686-4690, 1988). When protease is inactivated, immature viral particles are produced, which makes it an aim for effective treatment of AIDS.

Recent clinical studies involving human immunodeficiency virus protease inhibitors have reported significant therapeutic effects in AIDS patients, either alone or in combination with other drugs, such as reverse transcriptase inhibitors (Steele). , FP et al., Nature Med., 2: 257-258, 1996). Accordingly, various HIV protease inhibitors have been synthesized and as a result, a total of five protease inhibitors [indinavir (Crixivan ^™ , Merck); including amprenavir (Agenerase ^™ , Glaxo Wellcome), recently approved by the FDA; nefinavir from Viracept ^™ , Agouron; ritonavir (Norvir ^™ , Abbott); And saquinavir (Invirase ^™ , Hoffmann-La Roche) are used in the clinic. However, most of these drugs are peptidomimetic compounds, with the emergence of variant viruses with low oral bioavailability, fast release, complex synthesis steps, and cross-resistance to drugs. This has been reported and has the disadvantage of showing side effects that cause diabetes when taken for a long time.

Therefore, there is a constant need to develop new compounds that can overcome the disadvantages of compounds that mimic peptides.

Accordingly, the present inventors have diligently researched to develop new compounds that can overcome the disadvantages of compounds that mimic peptides, and as a result, 6-hydroxy-1,3-dioxin-4- is completely different from conventional AIDS therapeutics. As a result of using a non-peptidyl compound having an on-ring, it overcomes the problems such as low oral bioavailability and rapid release of a compound that mimics a conventional peptide and drug resistance, as well as excellent HIV suppression effect and It was confirmed that the toxicity is low, the present invention was completed.

After all, the main object of the present invention is to provide a non-peptidic compound having a 6-hydroxy-1,3-dioxin-4-one ring that inhibits the protease of human immunodeficiency virus (HIV).

Another object of the present invention is to provide a method for producing an electrical compound.

The present invention provides a nonpeptidic compound having a 6-hydroxy-1,3-dioxin-4-one ring of general formula (I) below, which is an inhibitor of HIV protease.

Where

R ₁ is a methyl group, ethyl group, propyl group, isopropyl group, butyl group, isobutyl group or

Is a t-butyl group;

R ₂ is methyl, ethyl, propyl, isopropyl, butyl, isobutyl,

t-butyl group, benzene group, 4-aminophenyl group, 4-cyanophenyl group, 4-nitro

Group, phenyl group, phenethyl group, 2- (4-pyridyl) ethyl group, 4-cyanobenzenesulfone

Amido group, 4-aminobenzene sulfon amido group, 4-nitrobenzene sulfon amido

Group, 4-fluoromethylbenzenesulfonamido group, 5-cyanopyridinesulfonami

Pottery, 5-aminopyridine sulfonamido group, 5-nitropyridine amido group

Is a 5-fluoromethylpyridinesulfonamido group;

R ₃ is methyl, ethyl, propyl, isopropyl, cyclopropyl, butyl

Group, isobutyl group, cyclobutyl group or t-butyl group; And,

R ₄ is a phenyl group, 4-aminophenyl group, 4-cyanophenyl group, 4-nitrophenyl group, 2-py

Lidine group, 5-cyanopyridine group, 5-aminopyridine group, 5-nitropyridine

Group, 5-trifluoromethylpyridine group or 1-methyl-4-imidazole group

to be.

Pharmaceutically acceptable salts of compounds of formula (I) of the invention include acetates, adipates, aspartates, benzoates, benzenesulfonates, sulfates, citrate, camphorates, camphorsulfonates, diphosphates, ethane Sulfonate, fumarate, glutamate, maleate, lactate, methanesulfonate, nitrate, succinate, tartrate, picrate or tosylate.

Since the compound of general formula (I) of the present invention has a chiral center, it is possible to form racemates, racemic mixtures or individual diastereomers, and enantiomers having all isomeric forms are within the scope of the present invention. Included.

The process for the preparation of the compound of general formula (I) having the 6-hydroxy-1,3-dioxin-4-one ring of the present invention is characterized in that the alkylbutylate (II) and the alkyl dialkylphosphonate (III) And reacting in the presence of butyl lithium to obtain β-ketophosphonate (IV); reacting β-ketophosphonate (IV) with NaH, followed by addition of aldehyde, followed by reduction in the presence of hydrogen and a catalyst to obtain ketone (V); Cyclizing the ketone (V) in the presence of sulfuric acid, acetic acid and malonic acid to obtain 6-hydroxy-1,3-dioxin-4-one (VI); Condensation of 6-hydroxy-1,3-dioxin-4-one (VI) with meta-nitrobenz aldehyde in the presence of argon and catalyst, and in the reaction solvent in the presence of bromo copper-dialkyl sulfide Reacting with ethyl aluminum to obtain a nitro compound (VII); Reducing the nitro compound (iii) in the presence of hydrogen and a catalyst to obtain an amine compound (iii); And a step of reacting the amine compound (iii) with allylsulfone chloride in the presence of a pyridine base to produce a compound of general formula (I).

Hereinafter, a method for preparing a compound of formula (I) having a 6-hydroxy-1,3-dioxin-4-one ring of the present invention will be described in detail by dividing step by step.

First step : obtaining β-ketophosphonate (IV)

Alkyl butyrate (II) and alkyl dialkylphosphonate (III) are reacted in the presence of nitrogen and butyl lithium to obtain β-ketophosphonate (IV): wherein the reaction solvent is an organic solvent such as THF , Reaction temperature is -80 to -70 ℃, reaction time is 3 to 5 hours.

Second Step : Obtaining Ketone (V)

reacting β-ketophosphonate (IV) with NaH, followed by addition of aldehyde, followed by reduction in the presence of hydrogen and a catalyst to obtain ketone (V); At this time, the reaction solvent is an organic solvent such as THF, Pd / C can be used as a catalyst, the amount of catalyst added is 5 to 15% (w / w) of the reactants, the reduction reaction solvent is methyl alcohol, ethyl alcohol, And lower alcohols such as propyl alcohol, butyl alcohol and pentyl alcohol.

Third Step : Obtaining 6-hydroxy-1,3-dioxin-4-one (VI)

Ketone (V) is cyclized in the presence of sulfuric acid, acetic acid and malonic acid to give 6-hydroxy-1,3-dioxin-4-one (VI).

Fourth Step : Obtaining a Nitro Compound

Condensation of 6-hydroxy-1,3-dioxin-4-one (VI) with meta-nitrobenz aldehyde in the presence of argon and catalyst and tri-reaction in the reaction solvent in the presence of bromo copper-dialkyl sulfide Reaction with ethyl aluminum yields the nitro compound (VII); In this case, the catalyst is preferably aluminum chloride, the reaction solvent is an organic solvent such as THF.

Fifth Step : Obtaining an Amine Compound

The nitro compound (VII) is reduced in the presence of hydrogen and a catalyst to obtain an amine compound (IX); In this case, Pd / C may be used as the catalyst, and the amount of catalyst added is 5 to 15% (w / w) of the reactants, and the reaction solvent is a lower level such as methyl alcohol, ethyl alcohol, propyl alcohol, butyl alcohol, pentyl alcohol, and the like. Alcohol.

Step 6 : Preparation of a Compound of Formula (I)

The amine compound (VII) is reacted with allylsulfone chloride in the presence of a pyridine base to prepare a compound of formula (I).

Where

R ₁ , R ₂ , R ₃ and R ₄ are the same as already defined.

The compound having a 6-hydroxy-1,3-dioxin-4-one ring of the present invention showed an excellent inhibitory effect on the protease, showing a low cytotoxicity.

Hereinafter, the present invention will be described in more detail with reference to Examples. These examples are only for illustrating the present invention in more detail, it will be apparent to those of ordinary skill in the art that the scope of the present invention is not limited by these examples in accordance with the gist of the present invention. .

Example 1 N- {3- [1- (6-hydroxy-4-oxo-2-phenethyl-2-propyl-4H- [1,3] dioxin-5-yl) propyl] phenyl}- Preparation of 2- (5-trifluoromethylpyridine) sulfonamide

Example 1-1 Obtaining Dimethyl Butanomethylphosphonate

After dissolving 8.6 ml of dimethyl methylphosphonate (79 mmol) in 40 ml of THF and cooling to -78 ° C, 60 ml of n-butyl lithium (1.33 M, 79 mmol) was slowly added under nitrogen atmosphere, followed by stirring for 30 minutes. The solution of methyl butyrate (4.5 ml, 39 mmol) dissolved in 20 ml of THF was added slowly, stirred for 4 hours, gradually raised to room temperature at -78 ° C, and the reaction was terminated with 2N hydrochloric acid. The THF solvent was removed, the organic layer was extracted with dichloromethane in the aqueous layer, and then dried and purified to give the title compound (6.6 g, 87%).

¹ H NMR (CDCl ₃ , 330 MHz): δ3.81 (s, 3H), 3.77 (s, 3H),

3.10 (d, 2H, J = 22.6 Hz),

2.61 (t, 2H, J = 7.2 Hz),

1.64 (q, 2H, J = 3.7 Hz),

0.94 (t, 3H, J = 7.4 Hz).

Example 1-2 : Obtaining 1-phenyl-1-hexan-3-one

100 ml of THF solution containing 60% NaH (1.6 g, 31.5 mmol) was cooled to 0 ° C. and dimethyl butanomethylphosphonate (4.1 g, 21 mmol) dissolved in 40 ml of THF under nitrogen atmosphere was added. . The reaction mixture was slowly raised to room temperature, left to stand for one hour, and then benzaldehyde (3.34 g, 3.5 mmol) was added and stirred for 30 minutes, and then, water was added to the reaction mixture to terminate the reaction. Subsequently, the organic layer was extracted with ether, dried and the obtained residue was purified to give 1-phenyl-1-hexen-3-one (3.4 g, 93%).

¹ H NMR (CDCl ₃ , 330 MHz): δ 7.79-7.39 (m, 6H), 6.76 (d, 1H, J = 16.3 Hz),

2.67 (t, 2H, J = 7.4 Hz),

1.68 (qt, 2H, J = 7.6, 7.4 Hz),

0.99 (t, 3H, J = 7.6 Hz).

10% Pd / C was added to 1-phenyl-1-hexen-3-one (560 mg, 3.21 mmol) dissolved in 30 ml of ethyl alcohol, stirred for 3 hours, and then the metal catalyst was filtered off to remove the filtrate. The residue obtained by concentration to give the title compound (560 mg, 92%).

¹ H NMR (CDCl ₃ , 330 MHz): δ 7.31-7.17 (m, 5H), 2.91 (t, 2H, J = 7.4 Hz),

2.73 (t, 2H, J = 7.5 Hz),

2.37 (t, 2H, J = 7.1 Hz),

1.60 (qt, 2H, J = 7.4, 7.1 Hz),

0.90 (t, 3H, J = 7.4 Hz).

Example 1-3 : Obtaining 2-phenethyl-2-propyl- [1,3] dioxane-4,6-dione

To 1-phenyl-1-hexan-3-one (500mg, 2.87mmol) and malonic acid (5.4g, 51.7mmol) dissolved in 6ml of acetic anhydride, 670mg of molecular sieve and 0.1ml of sulfuric acid were added. Stirred for 2 h. Subsequently, excess acetic anhydride was removed and the residue obtained was dispersed in iced water. The organic layer was extracted with ether, dried and concentrated to obtain the title compound (243 mg, 32%).

¹ H NMR (CDCl ₃ , 300 MHz): δ 7.32-7.18 (m, 5H), 3.63 (s, 2H),

2.89-2.77 (m, 2H), 2.27-2.21 (m, 2H),

1.99-1.94 (m, 2H), 1.58-1.52 (m, 2H),

1.00 (t, 3H, J = 7.4 Hz);

IR (KBr): 2964, 1748 cm ^-1

Example 1-4 : Obtaining 6-hydroxy-5- [1- (3-nitrophenyl) propyl] -2-phenethyl-2-propyl- [1,3] dioxin-4-one

Under argon gas, aluminum chloride (247 mg, 1.865 mmol) is added to 10 ml of THF solution at -78 ° C, and the temperature is raised to room temperature. This solution was prepared by dissolving 2-phenethyl-2-propyl- [1,3] dioxane-4,6-dione (243 mg, 0.926 mmol) and meta-nitrobenzaldehyde (168 mg, 1.11 mmol) in 10 ml of THF solvent. The solution was added and stirred at room temperature for 1.5 hours. Subsequently, 1.12 g of sodium carbonate hydrate was added to terminate the reaction, and celite and magnesium sulfate were added, followed by stirring for 10 minutes. The reaction mixture was filtered, washed with ether, and the filtrate was concentrated under reduced pressure and dissolved in 10 ml of THF at 0 ° C. Then, Cu (I) Br-Me ₂ S (63 mg, 0.307 mmol) and triethyl aluminum (140 ml, 1,0M hexane solution) were added and stirred for 20 minutes, and water and 6N hydrochloric acid were added to terminate the reaction. Then, the residue obtained by drying and concentrating the organic layer extracted with ether was purified to give the title compound (185 mg, 47%).

¹ H NMR (CDOD ₃ , 300 MHz): δ 8.10 (s, 1H), 8.03 (m, 1H),

7.62 (d, 1 H, J = 7.6 Hz), 7.43 (m, 1 H),

7.16-6.97 (m, 4H), 6.91-6.89 (d, 1H, J = 6.9 Hz),

3.69 (m, 1H), 2.60 (m, 1H), 2.21-1.91 (m, 4H),

1.88-1.80 (m, 2H), 1.50-0.95 (m, 3H),

0.86-0.62 (m, 6 H);

IR (KBr) 3226, 2966, 1740, 1458, 1286, 1172, 700 cm ^-1

Example 1-5 : Obtaining 6-hydroxy-5- [1- (3-aminophenyl) propyl] -2-phenethyl-2-propyl- [1,3] dioxin-4-one

6-hydroxy-5- [1- (3-nitrophenyl) propyl] -2-phenethyl-2-propyl- [1,3] dioxin-4-one (125 mg, 0.294 mmol) in 10 ml methanol It was dissolved and 75 mg of 10% Pd / C was added and stirred for 40 minutes under hydrogen atmosphere. The reaction mixture was filtered and the residue obtained by concentrating the filtrate was purified to give the title compound (115 mg, 99%).

¹ H NMR (CDOD ₃ , 300 MHz): δ 7.41-7.00 (m, 7H), 6.99-6.55 (m, 2H),

3.64 (m, 1H), 2.69 (m, 1H), 2.23-1.85 (m, 6H),

1.80-1.05 (m, 3H), 0.98-0.0.65 (m, 6H)

Example 1-6 N- {3- [1- (6-hydroxy-4-oxo-2-phenethyl-2-propyl-4H- [1,3] dioxin-5-yl) propyl] phenyl } -2- (5-trifluoromethylpyridine) sulfonamide (10d) Preparation

6-hydroxy-5- [1- (3-aminophenyl) propyl] -2-phenethyl-2-propyl- [1,3] dioxin-4-one (15 mg, 0.038 mmol) and pyridine (3.61 mg , 0.046 mmol) was dissolved in 1 ml of dichloromethane, and 4-trifluoromethylpyridine sulfonyl chloride (11.2 mg, 0.046 mmol) was added thereto and reacted with stirring at room temperature for 1.2 hours. Then, the residue obtained by concentrating the reaction product was purified to obtain the title compound (16 mg, 70%).

¹ H NMR (CDOD ₃ , 300 MHz): δ8.95-8.82 (m, 1H), 8.28-8.12 (m, 1H),

8.07-7.97 (m, 1 H), 7.15-6.80 (m, 9 H),

3.21 (m, 1H), 2.52 (m, 1H), 2.10-1.90 (m, 3H),

1.80-1.68 (m, 2H), 1.45-1.41 (m, 2H),

1.00-0.90 (m, 1H), 0.83-0.45 (m, 7H)

IR (KBr) 3256, 2968, 1742, 1176, 700 cm ^-1

Example 2 : N- {3- [1- (6-hydroxy-4-oxo-2-phenethyl-2-propyl-4H- [1,3] dioxin-5-yl) propyl] phenyl}- Preparation of (4-cyanophenyl) sulfonamide (10a)

Using a method similar to Example 1 above, the title compound was prepared (yield = 52%).

¹ H NMR (CDOD ₃ , 300 MHz): δ 7.85-7.65 (m, 3H), 7.63 (d, 1H, J = 8.6 Hz),

7.16-6.78 (m, 9H), 3.21 (m, 1H), 2.73 (M, 1H),

2.10-1.98 (m, 3H), 1.83-1.74 (m, 2H),

1.49-1.20 (m, 2H), 1.04 (m, 1H), 0.86-0.45 (m, 7H)

IR (KBr) 3266, 2966, 1738, 1284, 700 cm ^-1

Example 3 : N- {3- [1- (6-hydroxy-4-oxo-2-phenethyl-2-propyl-4H- [1,3] dioxin-5-yl) propyl] phenyl}- Preparation of Pyridinesulfonamide (10b)

Using a method similar to Example 1 above, the title compound was prepared (yield = 37%).

¹ H NMR (CDOD ₃ , 300 MHz): δ 8.59-8.48 (m, 1H), 7.89-7.78 (m, 2H),

7.47-7.16 (m, 1 H), 7.15-6.77 (m, 9 H),

3.38 (m, 1 H), 2.55 (m, 1 H), 2.10-1.87 (m, 4H),

1.74 (m, 1 H), 1.46 (m, 1 H), 1.30 (m, 1 H),

0.94 (m, 1 H), 0.85-0.45 (m, 7 H)

IR (KBr) 3460, 2966, 1742, 1178, 700 cm ^-1

Example 4 : N- {3- [1- (6-hydroxy-4-oxo-2-phenethyl-2-propyl-4H- [1,3] dioxin-5-yl) propyl] phenyl}- Preparation of 2- (5-cyanopyridine) sulfonamide (10c)

Using a method similar to Example 1 above, the title compound was prepared (yield = 50%).

¹ H NMR (CDOD ₃ , 300 MHz): δ8.96-0.74 (m, 1H), 8.31-8.12 (m, 1H),

8.03-7.90 (m, 1 H), 7.15-6.90 (m, 9 H),

3.38 (m, 1 H), 2.55 (m, 1 H), 2.06-1.97 (m, 3 H),

1.82-1.71 (m, 2H), 1.47 (m, 1H), 1.31 (m, 1H),

0.97 (m, 1 H), 0.85-0.49 (m, 7 H)

IR (KBr) 3440, 1640, 700 cm ^-1

Example 5 : N- {3- [1- (6-hydroxy-4-oxo-2-phenethyl-2-propyl-4H- [1,3] dioxin-5-yl) propyl] phenyl}- Preparation of 2- (5-nitropyridine) sulfonamide (10e)

Using a method similar to Example 1 above, the title compound was prepared (yield = 51%).

¹ H NMR (CDOD ₃ , 300 MHz): δ9.42-9.22 (m, 1H), 8.70-8.50 (m, 1H),

8.11-7.96 (m, 1 H), 7.18-6.80 (m, 9 H),

3.37 (m, 1 H), 2.47 (m, 1 H), 2.07-1.90 (m, 3 H),

1.79-1.97 (m, 2H), 1.47 (m, 1H), 1.26 (m, 1H),

1.8 (m, 1 H), 0.97-0.50 (m, 7 H)

IR (KBr) 3428, 2966, 1742, 1180, 700 cm ^-1

Example 6 : N- {3- [1- (6-hydroxy-4-oxo-2-phenethyl-2-propyl-4H- [1,3] dioxin-5-yl) propyl] phenyl}- Preparation of 2- (5-aminopyridine) sulfonamide (10f)

Using a method similar to Example 1 above, the title compound was prepared (yield = 94%).

¹ H NMR (CDOD ₃ , 300 MHz): δ 7.87-7.76 (m, 1 H), 7.61-7.45 (m, 1 H),

7.15-6.73 (m, 10 H), 3.38 (m, 1 H), 2.55 (m, 1 H),

2.17-1.65 (m, 5H), 1.30- 1.15 (m, 3H),

0.85-0.46 (m, 7H)

Example 7 : N- {3- [1- (6-hydroxy-4-oxo-2-phenethyl-2-propyl-4H- [1,3] dioxin-5-yl) propyl] phenyl}- Preparation of 4- (1-methylimidazole) sulfonamide (10 g)

Using a method similar to Example 1 above, the title compound was prepared (yield = 30%).

¹ H NMR (CDOD ₃ , 300 MHz): δ 7.51-6.80 (m, 11H), 3.52 (1H, m), 3.33 (s, 3H),

2.65 (m, 1H), 2.20-1.53 (m, 8H), 1.41-0.84 (m, 7H)

IR (KBr) 3400, 1734, 1274, 700 cm ^-1

Example 8 HIV-1 Protease Inhibitory Effect

First, the compounds prepared in Examples 1 to 7 were dissolved at the same concentration in experimental buffer solution (50 mM sodium acetate, 200 mM NaCl, 5 mM DTT and 10% glycerol, pH 5.2). 9 µl of the electrochemical solution, 6.5 µl of the substrate (Abz-Thr-Ile- p- nitro-Phe-Gln-Arg-NH ₂ (Bachem Bioscience)) dissolved in 100% DMSO at 1 mg / ml concentration, 72.5 µl of experimental buffer solution And 2 μl of HIV-1 protease were mixed, and degradation reaction of the substrate by the protease was performed at room temperature for 1 hour. Subsequently, the highest emission wavelength fluorescence of 460 nm according to the degree of substrate decomposition was measured using a fluorescence microplate analyzer (Fluoroskan, Labsystems). As a result of the experiment, the concentration that inhibits the protease activity by 50% is expressed as IC ₅₀ , and the lower the value, the better the anti-protease effect.

Example 9 Cytotoxicity and Antiviral Measurement

After centrifugation of MT-4 cells, the supernatant was removed and the cell sediment was inoculated with virus (HIV-1 strain III _B ) in an amount of 100 CCID ₅₀ (50% cell proliferation inhibitory dose). 10% RPMI 1640 was added and diluted to 2 × 10 ⁵ cells / ml. Meanwhile, AZT (azidothymidine) as a control and a compound prepared in Examples 1 to 7 were dissolved in 100% DMSO at a concentration of 20 mg / ml, and diluted twice with a culture solution containing RPMI 1640 and 10% FBS. 100 μl / well was dispensed into the well plate, and then 100 μl / well of the electric cell diluent was dispensed into the well plate, and cultured in a 37 ° C. carbon dioxide incubator for 5 days. After incubation, the toxicity and efficacy of the samples were evaluated by MTT screening method. That is, the toxicity of the drug was determined by determining CC ₅₀ (50% cytptoxic concentration) of the drug that caused 50% of the cells to die by comparing the number of cells surviving in each experimental group that was not inoculated with the virus. The antiviral effect was measured by determining the concentration of the drug that allowed 50% of the infected cells to survive as EC ₅₀ (50% effective concentration).

Cytotoxicity (%) = [1- (A _T ) _mock ] / [(Ac) _mock ] × 100

Antiviral effect (%) = [(A _T ) _HIV- (A _C ) _HIV ] / [(Ac) mock- (A _C ) _HIV ] × 100

Where

(A _C ) _mock is the absorbance of wells without added virus or drug;

(A _T ) _mock is the absorbance of the well to which the drug is added;

(A _T ) _HIV is the absorbance of the virus and drug addition wells; And,

(A _C ) _HIV is the absorbance of the virus-only wells.

As a result of the experiment, the higher the survival rate at each concentration, the less virulence of the sample itself, and the antiviral effect can be judged to be greater. In cytotoxicity, if the drug is not toxic at all, it will have a 100% survival rate, and if it is strong enough to kill all cells it will have a 0% survival rate. The concentration of the drug that kills 50% of the cells is expressed as CC ₅₀ , with higher values indicating less toxicity. The concentration of drug that the sample can exhibit the maximum antiviral effect is calculated by EC _50. The lower the value, the better the antiviral effect. Further, to the ratio of CC ₅₀ and EC ₅₀ that the selectivity (SI), may be the higher the value.

Table 1 shows the results of HIV-1 protease inhibitory effect, antiviral activity, cytotoxicity and selectivity (SI) of the compounds prepared in Examples 1 to 7. As a control material, tipranavir (PNU-140690) (Compound 2) was synthesized and used in a known manner.

HIV-1 Protease Inhibitory and Antiviral Effects of the Compounds of the Present Invention compound IC ₅₀ (μM) EC ₅₀ (μM) CC ₅₀ (μM) SI (CC ₅₀ / EC ₅₀ ) 10a 0.015 66.53 66.53 <1 10b 0.013 86.71 178 2.05 10c 0.003 11.37 76.96 6.77 10d 0.010 0.96 63.01 65.69 10e 0.065 14.79 14.79 <1 10f 0.013 69.97 69.97 <1 10 g 0.001 83.57 83.57 <1 Compound 2 0.050 0.70 18.60 26.57

As shown in Table 1, the inhibitory effect on the HIV-1 protease enzyme of the compounds having a 6-hydroxy-1, 3-dioxin-4-one ring of the present invention prepared in Examples 1 to 7 (IC ₅₀ = 0.001 to 0.065 uM), it can be seen that the compounds of the present invention are generally superior to the compound 2 (IC ₅₀ = 0.050 uM). The antiviral activity (EC ₅₀ ) using MT-4 cells was lower than that of the control, but compound 10d (EC ₅₀ = 0.96 uM) showed equivalent activity. In the case of cytotoxicity, except for compound 10e, the compounds of the present invention showed a significantly lower cytotoxicity (CC ₅₀ = 66.01 to 178uM) than the control compound 2 (CC ₅₀ = 18.60uM), it was found to be a safe drug. In particular, the selectivity (SI) by the ratio of the CC ₅₀ and EC ₅₀ showing the therapeutic effect of the drug was found that the compound 10d (SI = 65.69) is more than twice as effective as the control (SI = 26.57).

As described and demonstrated in detail above, the present invention provides a non-peptidic compound having a 6-hydroxy-1,3-dioxin-4-one ring that inhibits protease, an essential enzyme for human immunodeficiency virus (HIV), and It provides a manufacturing method. Non-peptidic compounds having a 6-hydroxy-1,3-dioxin-4-one ring of the present invention are excellent in the inhibition of HIV protease and antiviral activity, and markedly low in cytotoxicity, thereby treating AIDS or HIV infection. Or it may be usefully used in the development of a therapeutic for prevention.

As described above in detail the specific parts of the present invention, for those skilled in the art, such a specific description is only a preferred embodiment, which is not to limit the scope of the present invention by this Will be obvious. Accordingly, the substantial scope of the present invention will be defined by the appended claims and the like and their equivalents.

Claims (16)

Compounds having 6-hydroxy-1,3-dioxin-4-one rings represented by the following general formula (I), isomers thereof and pharmaceutically acceptable salts thereof: Where R ₁ is a methyl group, ethyl group, propyl group, isopropyl group, butyl group, isobutyl group or Is a t-butyl group; R ₂ is methyl, ethyl, propyl, isopropyl, butyl, isobutyl, t-butyl group, benzene group, 4-aminophenyl group, 4-cyanophenyl group, 4-nitro Group, phenyl group, phenethyl group, 2- (4-pyridyl) ethyl group, 4-cyanobenzenesulfone Amido group, 4-aminobenzene sulfon amido group, 4-nitrobenzene sulfon amido Group, 4-fluoromethylbenzenesulfonamido group, 5-cyanopyridinesulfonami Pottery, 5-aminopyridine sulfonamido group, 5-nitropyridine amido group Is a 5-fluoromethylpyridinesulfonamido group; R ₃ is methyl, ethyl, propyl, isopropyl, cyclopropyl, butyl Group, isobutyl group, cyclobutyl group or t-butyl group; And, R ₄ is a phenyl group, 4-aminophenyl group, 4-cyanophenyl group, 4-nitrophenyl group, 2-py Lidine group, 5-cyanopyridine group, 5-aminopyridine group, 5-nitropyridine Group, 5-trifluoromethylpyridine group or 1-methyl-4-imidazole group to be. The method of claim 1, Pharmaceutically acceptable salts of compounds represented by general formula (I) are acetate , Adipate, aspartate, benzoate, benzenesulfonate, sulfate , Citrate, camphorrate, camphorsulfonate, diphosphate, ethanesulphate Phonates, Fumarates, Glutamate, Maleates, Lactates, Methanesulfo Nate, Nitrate, Succinate, Tartrate, Picrate or Sat Characterized in that the Compounds having 6-hydroxy-1,3-dioxin-4-one rings, isomers thereof and pharmaceutically acceptable salts thereof. The method of claim 1 Inhibiting the activity of protease of human immunodeficiency virus Characterized Compounds having 6-hydroxy-1,3-dioxin-4-one rings, isomers thereof and pharmaceutically acceptable salts thereof. N- {3- [1- (6-hydroxy-4-oxo-2-phenethyl-2-propyl-4H- [1,3] dioxin-5-yl) propyl] phenyl}-(4-sia Nophenyl) sulfonamide, isomers thereof and pharmaceutically acceptable salts thereof. N- {3- [1- (6-hydroxy-4-oxo-2-phenethyl-2-propyl-4H- [1,3] dioxin-5-yl) propyl] phenyl} -pyridinesulfonamide, Isomers thereof and pharmaceutically acceptable salts thereof. N- {3- [1- (6-hydroxy-4-oxo-2-phenethyl-2-propyl-4H- [1,3] dioxin-5-yl) propyl] phenyl} -2- (5 -Cyanopyridine) sulfonamide, isomers thereof and pharmaceutically acceptable salts thereof. N- {3- [1- (6-hydroxy-4-oxo-2-phenethyl-2-propyl-4H- [1,3] dioxin-5-yl) propyl] phenyl} -2- (5 -Trifluoromethylpyridine) sulfonamide, isomers thereof and pharmaceutically acceptable salts thereof. N- {3- [1- (6-hydroxy-4-oxo-2-phenethyl-2-propyl-4H- [1,3] dioxin-5-yl) propyl] phenyl} -2- (5 -Nitropyridine) sulfonamides, isomers thereof and pharmaceutically acceptable salts thereof. N- {3- [1- (6-hydroxy-4-oxo-2-phenethyl-2-propyl-4H- [1,3] dioxin-5-yl) propyl] phenyl} -2- (5 -Aminopyridine) sulfonamides, isomers thereof and pharmaceutically acceptable salts thereof. N- {3- [1- (6-hydroxy-4-oxo-2-phenethyl-2-propyl-4H- [1,3] dioxin-5-yl) propyl] phenyl} -4- (1 -Methylimidazole) sulfonamide, isomers thereof and pharmaceutically acceptable salts thereof. (Iii) reacting alkylbutylate (II) and alkyl dialkylphosphonate (III) in the presence of nitrogen and butyl lithium to obtain β-ketophosphonate (IV); (Ii) reacting β-ketophosphonate (IV) with NaH, reacting with addition of aldehyde, and then reducing in the presence of hydrogen and a catalyst to obtain ketone (V); (Iii) cyclizing the ketone (V) in the presence of sulfuric acid, acetic acid and malonic acid to obtain 6-hydroxy-1,3-dioxin-4-one (VI); (Iii) 6-hydroxy-1,3-dioxin-4-one (VI) is condensed with meta-nitrobenz aldehyde in the presence of argon and catalyst and in the presence of bromo copper-dialkyl sulfide Reacting with triethyl aluminum in a solvent to obtain a nitro compound (VII); (Iii) reducing the nitro compound (iii) in the presence of hydrogen and a catalyst to obtain an amine compound (iii); And, (Iii) 6-hydroxy-1,3-dioxin-4-one comprising the step of reacting an amine compound (iii) with allylsulfone chloride in the presence of a pyridine base to produce a compound of formula (I) Method for preparing a compound having a ring: Where R ₁ , R ₂ , R ₃ and R ₄ are the same as already defined. The method of claim 11, The catalyst of the process (ii) is Pd / C, and the addition amount is 5-15% (w / w) of the reactants. Characterized by Process for the preparation of a compound having a 6-hydroxy-1,3-dioxin-4-one ring. The method of claim 11, (Ii) Reduction reaction of the process is methyl alcohol, ethyl alcohol, propyl alcohol, butyl alcohol Or in the solvent of pentyl alcohol. Process for the preparation of a compound having a 6-hydroxy-1,3-dioxin-4-one ring. The method of claim 11, (Iii) the catalyst of the process is characterized in that the aluminum chloride Process for the preparation of a compound having a 6-hydroxy-1,3-dioxin-4-one ring. The method of claim 11, The catalyst of step (iii) is Pd / C, and the addition amount is 5-15% (w / w) of the reactants. Characterized by Process for the preparation of a compound having a 6-hydroxy-1,3-dioxin-4-one ring. The method of claim 11, (Iii) Reduction reaction of the process is methyl alcohol, ethyl alcohol, propyl alcohol, butyl alcohol Or in the solvent of pentyl alcohol. Process for the preparation of a compound having a 6-hydroxy-1,3-dioxin-4-one ring.