HK1016601B - Furan-and thiophenecarbothioamide derivatives, their preparation and their use as inhibitors of the replication of hiv-1 and hiv-1 mutants - Google Patents

Description

Furan-and thiophenecarboxamide derivatives, their preparation and their use as inhibitors of HIV-1 and HIV-1 mutant replication

The present invention relates to compounds useful for inhibiting replication of HIV-1 and HIV-1 mutants. More particularly, the present invention relates to methylfuryl-and methylthiophene-pentenyl ether derivatives which are inhibitors of replication of wild-type HIV-1 and HIV-1 reverse transcriptase mutants. The invention also relates to a method for preventing or treating HIV-1 infection in a patient, which comprises administering to the patient an effective amount of said methylfuryl-or methylthiophen-pentenyl ether derivative.

Various compounds acting on virally encoded Reverse Transcriptase (RT), e.g., nevirapine, pyridone (pyridone), TIBO, BHAP, TSAO and quinoxaline, have been disclosed as in vitro inhibitors of human immunodeficiency virus type 1 (HIV-1). US patent US5,268,389 discloses certain thiocarboxylate compounds useful for inhibiting HIV replication. The selectivity of the compound for HIV-1 is due to a highly specific interaction with HIV-1 RT.

The rapid development of HIV-1 strains resistant to several HIV-1-specific RT inhibitors in cell cultures and in AIDS patients has raised concerns for the further development of such inhibitors in the clinic. For example, HIV-1 strains that contain a 100Leu → Ile mutation in their RT are anti-TIBO R82913 and R82150. anti-TSAO derivatives of HIV-1 strain containing a 138Glu → Lys mutation at its RT. The 181Tyr → Cys mutation in RT of the HIV-1 strain renders the mutant virus resistant to neyirapine and pyridone. See, for example, Balzarini et al, j.virology 67 (9): 5353-: 246, 253(1993) ("Balzarini II").

Combinations of various HIV-1RT inhibitors have been tried to overcome viral resistance. See, for example, Balzarini I.

De Clercq，E.，Int.J.Immunotherapy X(4): 145-158(1994) describe mutations in HIV-1RT which confer resistance to certain HIV-1 specific RT inhibitors.

WO96/16675 describes a composition for preventing and treating HIV-1 infection comprising an HIV-1RT inhibitory heterocyclylcarboxanilide or thiocarboxamide compound, a second HIV-1RT inhibitory compound which is not selected to be the same HIV-1RT mutant as the HIV-1RT inhibitory heterocyclylcarboxanilide or thiocarboxamide compound, and optionally a third HIV-1RT inhibitor compound.

It is an object of the present invention to provide compounds which themselves inhibit or suppress the emergence of wild-type HIV-1 and HIV-1RT mutants. It is also an object of the present invention to provide a method for preventing or treating HIV-1 infection by administering such compounds.

The present invention relates to compounds of the formula:wherein X is O or S.

The compounds of the present invention are useful for inhibiting the replication of human immunodeficiency virus-1 (HIV-1) and its Reverse Transcriptase (RT) mutants in vitro and in vivo. The compounds are useful in the treatment or prophylactic treatment of diseases caused by HIV-1 and mutants thereof of RT, such as acquired immunodeficiency syndrome (AIDS).

The invention also relates to a pharmaceutical composition comprising a therapeutically effective amount of a compound of formula I and a pharmaceutically acceptable carrier.

The present invention also relates to a method of treating HIV-1 infection in an infected host, which comprises administering to said host a therapeutically effective amount of a compound of formula I.

Desirably, the present invention relates to compounds of the formula:or

Synthesis method

By reacting an acid of the formula A-COOH, wherein A isWith aniline derivatives of the formulaReacted to prepare the compounds of the invention. The acid A-COOH is first converted to its acid chloride and then treated with the aniline derivative and an acid scavenger in a suitable solvent to form the amide. The acid scavenger may be an organic base such as pyridine, or an alkali metal hydroxide, carbonate or bicarbonate such as sodium bicarbonate. Suitable solvents for this step include: dichloromethane, diethyl ether, ethyl acetate, and the like. The resulting amide is then reacted with a sulfinylating agent such as Lawesson's reagent or phosphorus pentasulfide in a suitable solvent in the presence of an acid scavenger such as pyridine or sodium bicarbonate. To complete the thioacylation reaction, heating is generally carried out. Suitable solvents for the thionylation reaction include toluene, xylene, DME, and the like.[S]Arbitrary sulphurous acylating agents

A second method of preparing the compounds of the invention is to metallate the bromo compound of formula A-Br with n-butyl lithium at a temperature of-75 ℃ to-80 ℃. The resulting lithium complex is then reacted with an isothiocyanate of the formula:the compound of the invention (thiocarboxamide) can then be formed directly by acidification.

Control compounds 1-117 can be prepared in a similar manner to the above method. Typically, the preparation of the control compound involves the formation of an amide followed by thioacylation with Lawesson's reagent or phosphorus pentasulfide.

(i) Alkali

(ii) Lawesson's reagent or P₂S₅With pyridine or sodium bicarbonate

A. Z and B have the meanings given in Table 1.

The compounds of the present invention may be administered orally, parenterally, buccally, as an inhalation, rectally, or dermally in dosage unit formulations containing conventional non-toxic pharmaceutically acceptable carriers, adjuvants and vehicles. Pharmaceutically acceptable carriers, adjuvants and vehicles which may be employed in the compositions of the invention are found in standard Pharmaceutical texts, such as Remington's Pharmaceutical Sciences, 16th Edition, Mack Publishing Company, Easton, Pa (1980).

The therapeutically effective amount of a compound of the present invention which may be combined with the pharmaceutical carrier to make a single dosage form will vary with the age, condition and particular mode of administration of the subject host. In general, the compounds of the invention are preferably used in concentrations such that: generally, effective antiviral effects can be produced without causing any harmful side effects.

Although the compounds of the present invention may be used as the sole active agent, the compounds of the present invention may also be used in combination with one or more other agents which do not destroy the activity of the compounds of the present invention or whose combination with the present compounds does not produce an injurious effect on the host treated.

The following examples are provided to illustrate the invention.

Examples Materials and methods Test compounds

(A) Compound IA was prepared as follows: (method I) step 1Preparation of 2-chloro-5-nitrophenol

2-amino-5-nitrophenol (65.6g) was added to 36% hydrochloric acid (200ml) with stirring at 0-5 ℃. A solution of sodium nitrite (33.25g) in water (75ml) was added dropwise over 1.5 hours, and then the first reaction mixture was maintained at this temperature for a further 1 hour. Decomposition of the excess sulfoxide with sulfamic acid (1.5g) added in portionsNitric acid. The first reaction mixture was then added portionwise to a stirred suspension of the ketone (I) chloride (8.5g) dissolved in 20% hydrochloric acid (50 ml). A large amount of foam is generated. After the above addition process, the resulting second reaction mixture was stirred for 1 hour. The precipitate from the second reaction mixture was then collected on a filter, washed with water and dried to give 63.2g of a brown solid. The solid was refluxed with 1.5g of activated carbon in methanol (500ml) for 15 minutes; filtration through celite (celite); evaporation gave 60.8g of brown 2-chloro-5-nitrophenol. Step 2Preparation of 1-chloro-2- (3-methyl-2-butenyloxy) -4-nitrobenzene

A reaction mixture of 2-chloro-5-nitrophenol (50.6g), anhydrous potassium carbonate (44.5g), tetrabutylammonium bromide (4.7g) and 4-bromo-2-methyl-2-butene (53.3g, 90%) in methyl ethyl ketone (263ml) was stirred at ambient temperature overnight. TLC of the reaction mix showed a trace of phenol remaining. Brominated isoprene (1ml) was further added to the reaction mixture and stirred for 2 hours. The solvent is then removed from the reaction mixture. The residue of the reaction mixture was further treated with water and extracted into ether. With 2NNaOH₂And water washing the extract and drying (MgSO)₄) Filtered and evaporated to leave a brown solid which was recrystallized from ethyl acetate/isopropanol to give 1-chloro-2- (3-methyl-2-butenyloxy) -4-nitrobenzene, 49.5g as a beige solid, as a spot on TLC (ethyl acetate: hexane, 20: 80). Step 3Preparation of 4-chloro-3- (3-methyl-2-butenyloxy) aniline

To a well-stirred refluxing suspension of iron powder (19.6g, 100 mesh) in ethanol (60ml), water (13.4ml) and 36% hydrochloric acid (1.4ml) was added 1-chloro-2- (3-methyl-2-butenyloxy) -4-nitrobenzene (24g) in portions over 15-30 minutes. After 3 hours, Thin Layer Chromatography (TLC) (ethyl acetate: hexane, 40: 60) showed no substrate. The reaction mixture was then filtered hot and the iron oxide filter cake was washed with hot ethanol. The combined ethanol washes were evaporated to give a residue. The residue was dissolved in diethyl ether, washed with aqueous bicarbonate and water, and dried (MgSO)₄) Filtered and evaporated to obtain 4-chloro-3- (3-methyl-2)Butenyloxy) aniline, 19.8g of a light brown oil.

Step 4

Preparation of 2-methyl-3-furoyl chloride

A first reaction mixture of chloroacetaldehyde dimethanol (300g), water (400ml) and 36% hydrochloric acid (40ml) was stirred and refluxed. After the first reaction mixture was allowed to cool down and added to a stirred solution of ethyl acetoacetate (260g) and pyridine (500ml) and stirred at ambient temperature for 72 hours to give a second reaction mixture. The organic layer was then separated from the second reaction mixture, and the aqueous layer was diluted with water and extracted with dichloromethane. The combined organics were washed with 2N hydrochloric acid and then the solvent was removed. The residue was treated with a solution of water (700ml) and NaOH (80g) in ethanol (100ml) to give a third reaction mixture. After refluxing for 1 hour, the third reaction mixture was poured into ice/water and acidified with hydrochloric acid. A cream-colored precipitate formed. The precipitate was collected on a filter, washed with water and dried to obtain 180g of 2-methyl-3-furancarboxylic acid. 100g of 2-methyl-3-furancarboxylic acid was added in portions to thionyl chloride (500ml) and refluxed for 3 hours. The excess thionyl chloride was then distilled off to give a residue. The residue was distilled with a water pump to give 100g of 2-methyl-3-furoyl chloride, bp.62 ℃. Step 5Preparation of N- [ 4-chloro-3- (3-methyl-2-butenyloxy) phenyl]-2-methyl- 3-furancarboxamides

A first solution of 2-methyl-3-furoyl chloride (13.57g) in dichloromethane (94ml) was stirred in an ice/salt bath. A second solution of 4-chloro-3- (3-methyl-2-butenyloxy) aniline (19.8g) and triethylamine (14ml) in methylene chloride (94ml) was added to the first solution at such a rate as to maintain the temperature at-5 ℃ to 0 ℃. When the addition was complete, the resulting reaction mixture was stirred to ambient temperature and stirring was continued for 15 hours. Water was added to the reaction mixture, and then the organic layer was separated and washed with water, dilute hydrochloric acid, water, an aqueous sodium bicarbonate solution and water in this order. Drying (MgSO)₄) Thereafter removing the solvent from the washed organic layerAnd the residue was recrystallized from isopropanol to give 17g N- [ 4-chloro-3- (3-methyl-2-butenyloxy) phenyl-2-methyl-3-furancarboxamide as a white solid. A second crop of beige crystals was also obtained (7.6 g). Step 6Preparation of N- [ 4-chloro-3- (3-methyl-2-butenyloxy) -phenyl]-2-methyl -3-Furazthiocarboxamide (Compound IA)

11/2 hours later, the reaction solution is prepared from N- [ 4-chloro-3- (3-methyl-2-butenyloxy) phenyl]-2-methyl-3-furancarboxamide (4g), NaHCO₃A reaction mixture of (7.4g) and Lawesson's reagent (3.6g) in toluene (168ml) was gradually heated to 85 ℃ and held at that temperature for 21/2 hours. The reaction mixture was then allowed to cool and filtered through a neutral alumina packing eluting with diethyl ether to petroleum ether (1: 1). Evaporation of the solvent gave 2.6g N- [ 4-chloro-3- (3-methyl-2-butenyloxy) phenyl]-2-methyl-3-furancarbothioamide product.

(B) Compound IB was prepared as follows: (method II) step 1Preparation of 3, 5-dibromo-2-methylthiophene

Bromine (320g) in dioxane (2L) was added dropwise to 2-methylthiophene (98g) in dioxane (500ml) with stirring over 7.5 h, then allowed to stand overnight at ambient temperature. The reaction mixture was heated to reflux for another 3 hours, poured into water (4L) and extracted with ether. The ether extract was washed with aqueous bicarbonate solution and water, and dried (MgSO)₄). After evaporation, an oil was obtained which was distilled to give 234.6g of 3, 5-dibromo-3-methylthiophene, bp.98 ℃. Step 2Preparation of 3-bromo-2-methylthiophene

The magnesium turnings (5g) were covered with Tetrahydrofuran (THF) (25ml) and treated with 3, 5-dibromo-2-methylthiophene (5g) with stirring. An exotherm occurred. The reaction mixture was kept at a temperature of 35 ℃. + -. 5 ℃ by cooling. Additional 3, 5-dibromo-2-methylthiophene (47g) was added dropwise to the reaction mixture while maintaining the temperature within the above range. The temperature was raised to 40 ℃ towards the end of the addition. Most of the magnesium has reacted. After the reaction heat release is complete, the temperature of the reaction mixture is raised from 40 ℃ to 50 ℃ by external heating, andthe temperature was maintained at 50 ℃ for 1 hour. The reaction mixture was then slowly poured into vigorously stirred ice/water/dilute HCl to give a second mixture. The second mixture was extracted into ether, washed with water and dried (MgSO)₄) And evaporated to leave a liquid which is distilled in a water pump to give 32.6g of 3-bromo-2-methylthiophene, a clear liquid, bp.62-65 ℃. Step 3Preparation of 1-chloro-2- (3-methyl-2-butenyloxy) -4-isothiocyanatobenzene

1-chloro-2- (3-methyl-2-butenyloxy) aniline (19.9g) (see method I, step 3) was dissolved in dichloromethane (75ml) to form a first solution. The first solution was then covered with ice/water (250ml) under vigorous stirring. A second solution of thiophosgene (7.3ml) in dichloromethane (25ml) was then added dropwise to the first solution. The resulting reaction mixture was kept cool in an ice bath. A solid precipitated out of the reaction mixture, but over time the solid reacted further and was dissolved in dichloromethane. The reaction mixture was then allowed to return to ambient temperature at which point the reaction was complete. The organics were then separated from the reaction mixture, washed with water, dried and evaporated under reduced pressure (excess thiophosgene was absorbed with a caustic scrubber) to form a dark gray solid. The solid was purified by dissolving in commercial hexane, passing the dissolved solid through a column of silica gel (minimum 3.5cm diameter x 6cm length), and eluting the column with hexane. The hexane solution was evaporated to give 22g of a white solid as 1-chloro-2- (3-methyl-2-butenyloxy) -4-thiocyanatobenzene with one spot on TLC (hexane). Step 4Preparation of N- [ 4-chloro-3- (3-methyl-2-butenyloxy) -phenyl]-2-methyl -3-Thiophenylthiocarboxamide (Compound # IB)

A solution of 3-bromo-2-methylthiophene (5.5g) in dry diethyl ether (25ml) was cooled in an acetone/dry ice bath under nitrogen. N-butyllithium (10ml, 2M in hexane) was added dropwise to the solution at such a rate that the temperature of the resulting first reaction mixture was not increased more than 2 ℃. After this one-drop addition, the first reaction mixture was stirred at-75 ℃ for 1.5 hours. 1-chloro-2- (3-methyl-2-butenyloxy) -4-isocyano in dry diethyl ether (25ml)Sulfanylbenzene) (6.2g) was added dropwise to the first reaction mixture at such a rate that the temperature of the resulting second reaction mixture was not raised to 2 ℃ or higher (1.25 hours). After this addition process, the second reaction mixture was slowly returned to ambient temperature over 2 hours. The second reaction mixture was then treated with ice/water/dilute HCl/ether. Separating the organic from the second reaction mixture. The organics were washed with water and dried (MgSO)₄) And the solvent is removed. A viscous yellow oil remained. This oil gave a yellow solid when stirred with commercial hexane. The yellow solid was collected on the filter, 6.2g, and recrystallized from cyclohexane/diethyl ether to give 4.7g N- [ 4-chloro-3- (3-methyl-2-butenyloxy) phenyl]-2-methyl-3-thiophenecarboxamide, a yellow solid, mp118-120 ℃ (uncorrected).

Compounds IA and IB have satisfactory i.r. and NMR spectra.

TABLE 1Compound YZAB 1S Cl

2-methyl-3-

-OCH₂CH＝CHCH₃

Thienyl 2 "2-methyl-3-"

Furyl 3' -CH-N-O-C (CH)₃)₃4 ″ -OCH₃ ″ -CH＝N-O-CH₃5 ″ Cl

6-methyl-4, 5-

-CH＝N-O-C(CH₃)₃

dihydro-2H-6 "", a pharmaceutically acceptable salt thereof

Pyran-5-yl

-S-CH₂CH＝CHCH₃

2-methyl-3-

Furyl 7 ""

2-methyl-4, 5-

-CH＝N-O-C(CH₃)₃

Dihydro-3-furyl 8 "", a salt thereof, and a pharmaceutically acceptable salt thereof

2-methyl-3-S-CH₂CH＝CHCH₃

Thienyl 9 "", a10 ″ ″

1, 2-dimethyl-3-

-CH＝N-O-C(CH₃)₃

Pyrrole group 11 "

1-methyl-3- ", a salt thereof, and a pharmaceutically acceptable salt thereof

Pyrrolyl 12 "", a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable salt thereof

2-methyl-3-

-O-CH₂CH₂CH(CH₃)₂

Thienyl 13O ″)

5, 6-dihydro-2-ethyl-

1, 4-oxathiacyclohexadienes

-3-yl 14S ″

2-methyl-3-

-O-CH₂-C(O)-O-C(CH₃)₃

Thienyl 15O ″)

5, 6-dihydro-2-ethyl-

-CH＝N-O-C(CH₃)₃

1, 4-oxathiahexadienes

-3-yl 16S ″

2-methyl-3-

-C(O)-O-CH(CH₃)₂

Thienyl 17 "", a

2-methyl-3-

-S-CH₂-C(O)-O-C(CH₃)₃

Furyl radical

TABLE 1 (continuation)Compound Y Z A B18O Cl

5, 6-dihydro-2-methyl-

-CH＝N-O-C(CH₃)₃

1, 4-oxathiahexadienes

-3-yl 19S ″

1, 2-dimethyl-3-

-O-CH₂CH＝CHCH₃

Pyrrolyl 20 "", and its preparation method

2-methyl-3-

-S-CH₂-C(O)-O-C(CH₃)₃

Thienyl 21 "", a22 O ″

5, 6-dihydro-2-methyl-

1, 4-oxathiacyclohexadienes

-3-yl 23 ""

2-methyl-3-

Thienyl 24 "Cl

6-methyl-4, 5-

-CH＝N-O-C(CH₃)₃

dihydro-2H-

Pyran-5-yl 25S CN

2-methyl-3-

-O-CH₂-C(Cl)＝CH₂

Thienyl 26 "Cl

2-methyl-3-

Furyl 27O ″)

2-methyl-3-

-O-CH₂CH＝CH(CH₃)₂

Thienyl 28S ""29 ″ ″ ″ -OCH₂C(CH₃)₃

TABLE 1 (continuation)Compound Y Z A B30O Cl

2-methyl-3-

-OCH₂CH＝C(CH₃)₂

Furyl group 31 ""

2-methyl-3-

-C(O)-O-CH(CH₃)₂

Thienyl 32 "", a33 ″ ″ ″ -O-CH₂CH₂CH(CH₃)₂34 ″ ″ ″ -S-CH₂CH＝CHCH₃35 ″ ″

2-methyl-3-

-S-CH₂CH＝CHCH₃

Furyl 36 ""

2- (methylthio) -

-CH＝N-O-C(CH₃)₃

3-thienyl 37 ""

2-methyl-3-

-O-CH₂CH₂CH(CH₃)₂

Furyl 38 ""

1, 2-dimethyl-3-

-CH＝N-O-C(CH₃)₃

Pyrrolyl 39S ″)

2-methyl-3-

-O-CH₂C(CH₃)₃

Furyl 40O' -O-CH₂CH＝CHCH₃41 ″ ″

5, 6-dihydro-2-methyl-

-O-CH₂CH₂CH(CH₃)₂

1, 4-oxathiacyclohexadienes

-3-yl 42 ""

2-methyl-3-

Thienyl 43 ""

5, 6-dihydro-2-ethyl-

-O-CH₂CH＝CHCH₃

1, 4-oxathiahexadienes

-3-yl 44S ″

2-methyl-3-

-O-CH₂CH₂CH(CH₃)₂

Furyl 45O ″)

2-methyl-3-

-O-CH₂CH＝CHCH₃

Thienyl radical

TABLE 1 (continuation))Compound YZAB 46S Cl

2-methyl-3-

-O-C(O)-O-CH(CH₃)₂

Furyl 47O-OCH₃ ″ -CH＝N-O-CH₃48 O Cl ″ -C(O)-O-CH(CH₃)₂49 S ″ ″ ″50 O ″ ″ -C(O)-O-CH₂C(CH₃)＝CH₂51 S ″ ″ ″52 O ″ ″ -CH₂-O-C(CH₃)₃53 S ″ ″ ″54 O ″ ″

-C(O)-O-CH(CH₃)-

cyclo-C₃H₅55 S ″ ″ ″56 O -OCH₃ ″ -CH₂-C(O)-O-CH(CH₃)₂57 S ″ ″ ″58 O Cl

2-methyl-3-

Thienyl 59S "" 60O "" -CH ═ N-O-C (CH)₃)₃61S "" 62O "phenyl-C (O) -O-CH (CH)₃)₂63S "" 64O "2-fluorophenyl65S "" 66O "2-aminophenyl-C (O) -O-CH (C)₂H₅)₂67S "" 68O "2-methoxyphenyl69 S ″ ″ ″

TABLE 1 (continuation)Compound Y Z A B70OCl 2-methylphenyl radical71 S ″ ″ ″72 S Cl

2-methyl-3-

-CH＝N-O-CH₃

Furyl 73' -OCH₃ ″ -O-CH₂-CH＝CH₂74 ″ -CN ″ ″75 ″ -CH₃ ″ -C(O)-O-CH₂CH₃76 ″ Cl

3-methyl-2-

-O-CH₂-CH＝CH₂

Thienyl 77 ' -CN ' 78 ' -OCH₃"79" Cl 3-thienyl80 "" 2-chlorophenyl "81" "2-methoxyphenyl" 82 "" phenyl "83", and

2-methyl-3-

Furyl 84 "2-methylphenyl" 85 "" 2-aminophenyl-C (O) -O-CH (CH)₃)86 "2-furyl" 87 "" ″

3-methyl-2-

″

Furyl 88 ""

3-methyl-2- ", a pharmaceutically acceptable salt thereof

Thienyl 89 "" 2-thienyl "90" 2-methoxyphenyl "91" "2-methylphenyl" 92 "" 3-thienyl "93" "phenyl-CH ═ N-O-C (CH)₃)₃

TABLE 1 (continuation)Compound (I) Y Z A B94 S Cl

3-methyl-2-

-CH＝NOCH(CH₃)₂

Furyl 95 ""

3-methyl-2- ", a pharmaceutically acceptable salt thereof

Thienyl 96 "" phenyl "97" "2-methyl-3-"

Furyl 98' 2-fluorophenyl99 "" 2-pyrrolyl ″100 ″ ″

1-methyl-Z- ", a salt thereof, and a pharmaceutically acceptable salt thereof

Pyrrole group 101 ""

3-methyl-2- ", a pharmaceutically acceptable salt thereof

Thienyl 102 "", a

2-methyl-3- ", a pharmaceutically acceptable salt thereof

Furyl 103 "" -C (O) -O-CH₂CH(CH₃)₃104 ″ ″ ″ -C(O)-O-CH₂CH＝CH₂105 ″ ″ ″ -C(O)-O-CH₂-cyclo-C₃H₅106 ″ ″ ″ -C(O)-O-CH₂-CF₃107 ″ ″ ″ -C(O)-O-CH(CH₂CH₃)₂108 ″ ″ ″ -C(O)-O-CH₂CH(CH₂CH₃)₂109 ″ ″ ″ 110 ″ ″ ″

TABLE 1 (continuation)Compound YZAB 111S Cl

2-methyl-3-

-CH＝N-O-CH₂CH＝CH₂

Furyl group 112 "" - (CH-N-O-CH)₂C≡CH113 ″ ″ ″ 114 ″ ″ ″ -CH＝N-O-C(CH₃)₃115 ″ ″ ″ -O-CH₂-C(O)-O-C(CH₃)₃116 ″ ″ ″ -S-CH₂-C(O)-O-CH₂CH₃117 ″ ″ ″ -O-CH₂-C≡CHCells and viruses

CEM cells were obtained from the american tissue culture collection (Rockville, Md.). HIV-1 (III)_B) Culture supernatants originally obtained from H9 cells that persisted in infection with HIV-1, and were supplied by r.c. gallo and m.popovic (National cancer Institute, National Institutes of Health, Bethesda, MD). The selection and identification of HIV-1RT mutants was as follows: according to the method described by Balzarini et al (Virology)192: 246 (1993)), selecting HIV-1/100-Ile ("100-Ile") for resistance to TIBO R82150; according to the method described by Balzarini et al (Virology)192: 246-253(1993)) selecting HIV-1/103-Asn ("103-Asn") for resistance to TIBO R82913; the procedure described by Balzarini et al (J.Virol).67: 5353-5359(1993)) selecting HIV-1/106-Ala ("106-Ala") for resistance to nevirapine; according to the method described by Balzarini et al(Virology 192: 246, 253(1993) and Balzarini et al, Proc. Nat. Acad. Sci. USA90: 6952-6956(1993)) selection of HIV-1/Lys-138 ("Lys-138") vs TSAO-m³Resistance to T; according to the method described by Balzarini et al (Virology)192: 246-253(1993)) selecting for resistance of HIV-1/181-Cys ("181-Cys") to pyridone L-697, 661; the procedure described by Balzarini et al (mol. Pharmocol).44: 694 Astro 701(1993)) selecting for resistance of HIV-1/188-His ("188-His") to HEPT. Upon further passage in cell culture in the absence of HEPT, 188-His was further converted to HIV-1/188-Leu ("188-Leu").Testing antiviral Activity of Compounds in cell culture

CEM cells were suspended at a density of about 300,000 cells per ml of medium and used with about 100CCID₅₀(CCID₅₀50% cell culture infectious dose) of the above HIV-1 (III)_B) Or one of the HIV-1RT mutants. Then 100. mu.l of the infected cell suspension was added to the wells of a 200. mu.l microtiter plate containing 100. mu.l of appropriate serial (5-fold) dilutions of the test compound. The inhibitory effect of the test compounds on syncytia production in CEM cells induced by HIV-1 was examined microscopically on day 4 post-infection. 50% Effective Concentration (EC)₅₀) Is defined as the concentration of test compound that produces 50% inhibition of syncytia production in cultures of HIV-1 infected cells.Results

As can be seen from tables 2A and 2B below, compounds IA and IB have very low, virtually similar anti-HIV-1 (III)_B) (referred to as "WT" in tables 2A and 2B) and EC of HIV-1RT mutant₅₀The value is obtained. EC of anti-HIV-1 RT mutant strain of the two compounds₅₀The value is as low as 0.003 to 0.011 mu g/ml. This remarkable activity against the mutant HIV-1RT was surprising and unexpected (in comparison with the EC of the control compound 1-117 against the mutant HIV-1 RT)₅₀Values are comparative). Since the compounds of the present invention are effective in inhibiting wild-type HIV-1 and mutant strains of HIV-1RT, they will also be useful in inhibiting mutant strains of HIV-1RT which contain more than one mutation.

TABLE 2A

Antiviral activity ^c

Compound (I)	EC50(μg/ml)a							CC50(μg/ml)b
									WT	100-Ile	103-Asn	106-Ala	138-Lys	181-Cys	188-Leu
	IA	0.003	0.003	0.006	0.005	0.005	0.011									0.50	＞100
IB								0.003	0.004	0.004	0.005	0.005	0.005	0.60	5.8

^a50% effective concentration (i.e., the concentration of compound required to inhibit 50% of the viral-induced cellular morbidity)^b50% cell static concentration (i.e., the concentration of compound required to inhibit CEM cell proliferation by 50%)^cData are mean values from 2 to 3 independent experiments

TABLE 2B

Antiviral Activity of control Compounds ^c

Compound (I)	EC50(μg/ml)a							CC50(μg/ml)b
									WT	100-Ile	103-Asn	106-Ala	138-Lys	181-Cys	188-Leu
	1	0.0055	0.048	0.048	0.023	0.018	0.03									0.65	5.9
2								0.006	0.048	0.023	0.04	0.018	0.075	0.65	9.0
	3	0.0065	0.057	0.23	0.03	0.045	0.08									0.91	5.8
4								0.007	0.22	0.4	0.22	0.039	0.59	0.93	＞100
	5	0.013	0.055	0.08	0.025	0.04	0.025									＞2	10
6								0.0041	0.037	0.03	0.018	0.03	0.045	1.0	4.7
	7	0.0065	0.065	0.058	0.03	0.04	0.045									1.2	11
8								0.0055	0.032	0.023	0.023	0.025	0.025	≥2.0	5.1
	9	0.0032	0.057	0.13	0.03	0.02	0.032									＞2	6.0
10								0.02	0.65	1.27	0.23	0.09	0.033	＞2	6.5
	11	0.029	0.13	0.17	0.1	0.13	0.045									＞2	13
12								0.007	0.035	0.035	0.08	0.035	0.07	≥2.0	＞100
	13	0.08	0.26	0.38	0.17	0.25	0.07									＞2	11.4
14								0.067	0.16	0.12	0.1	0.24	0.087	＞2	2.1
	15	0.03	0.06	0.065	0.06	0.07	0.09									＞2	11.4
16								0.0041	0.15	0.33	0.077	0.03	0.1	≥2.0	5.9
	17	0.22	0.49	0.73	0.3	0.18	0.45									＞2	7.0
18								0.014	0.33	0.33	0.15	0.22	0.13	＞2	8.5
	19	0.17	0.85	≥2.0	0.4	0.6	0.13									＞2	5.2
20								0.13	0.34	0.44	0.15	0.43	0.14	＞2	5.4
	21	0.008	0.3	0.61	0.23	0.11	0.17									＞2	6.0
22								0.08	0.48	0.65	0.26	0.55	0.18	＞2	5.8
	23	0.03	0.7	≥2.0	0.22	0.73	0.27									＞2	5.8

TABLE 2B (continuation)

Antiviral Activity of control CompoundsProperty of (2) ^c

Compound (I)	EC50(μg/ml)a							CC50(μg/ml)b
									WT	100-Ile	103-Asn	106-Ala	138-Lys	181-Cys	188-Leu
	24	0.13	0.48	0.6	0.5	0.5	0.33									＞2	9.7
25								0.03	0.2	0.4	0.2	0.13	0.44	＞2	3.3
	26	0.033	0.33	0.33	0.09	0.50	≥2.0									＞2	5.3
27								0.01	0.36	0.5	0.11	0.15	0.46	＞2	6.0
	28	0.029	0.22	0.33	0.23	0.11	0.5									＞2	4.3
29								0.14	0.5	0.43	0.5	0.05	0.55	＞2	6.2
	30	0.015	1.27	1.67	0.34	0.23	0.65									＞2	＞100
31								0.09	≥2.0	≥2.0	0.6	0.93	0.73	＞2	5.5
	32	0.16	≥2.0	≥2.0	≥2.0	≥2.0	0.95									＞2	4.1
33								0.063	0.95	1.0	0.65	0.62	1.07	＞2	7.0
	34	0.08	0.65	0.8	0.28	0.5	1.2									＞2	4.8
35								0.16	1.5	0.55	0.8	0.7	1.6	＞2	5.9
	36	0.18	1.23	1.2	0.85	0.85	1.6									＞2	7.8
37								0.16	≥2.0	≥2.0	1.3	1.25	1.73	＞2	5.0
	38	0.08	≥2.0	≥2.0	0.7	≥2.0	＞2									＞2	5.5
39								0.15	1.4	0.7	＞2	0.5	≥2.0	＞2	4.1
	40	0.24	1.4	≥2.0	＞2	1.2	＞2									＞2	5.8
41								0.28	≥2.0	≥2.0	≥2.0	≥2.0	≥2.0	＞2	＞100
	42	0.36	≥2.0	≥2.0	≥2.0	≥2.0	≥2.0									＞2	5.0
43								0.65	1.2	1.3	4.5	1.3	＞2	＞2	11
	44	0.0073	0.035	0.08	0.06	0.03	0.08									＞10	＞100

TABLE 2B (continuation)

Antiviral Activity of control Compounds ^c

Compound (I)	EC50(μg/ml)a							CC50(μg/ml)b
									WT	100-Ile	103-Asn	106-Ala	138-Lys	181-Cys	188-Leu
	45	0.15	0.6	0.95	0.85	0.65	0.5									＞10.0	87
46								0.37	4.5	4.0	3.0	0.6	5.5	≥2	25
	47	0.23	≥10	≥10	≥10	≥10	≥10									≥10	＞100
48								0.45	6.5	-	6.5	3.5	6.0	-	3.8
	49	0.007	0.6	-	0.6	0.6	0.2									-	5.2
50								0.15	5.0	-	1.0	1.0	1.9	-	3.7
	51	0.006	0.8	-	0.03	0.03	0.10									-	6.7
52								0.70	8.5	-	5.5	4.0	≥10	-	6.2
	53	0.03	0.85	-	0.29	0.08	0.55									-	18
54								0.6	＞10	-	5.0	＞10	1.0	-	44
	55	0.009	0.65	-	0.08	0.18	0.11									-	17
56								0.7	＞10	-	6.5	＞10	≥10	-	52
	57	0.009	1.3	-	0.2	0.08	0.50									-	＞100
58								0.19	5.5	-	7.0	6.0	5.0	-	6.5
	59	0.09	0.65	-	0.60	0.40	0.35									-	15
60								0.23	4.5	-	1.6	1.57	3.0	-	14
	61	0.01	0.2	-	0.2	0.05	0.2									-	12
62								0.4	3.5	-	5.0	5.0	5.5	-	6.2
	63	0.05	4.5	-	5.0	2.2	5.0									-	4.3
64								0.6	≥10	-	2.0	4.0	5.0	-	33
	65	0.04	0.3	-	0.26	0.09	0.25									-	19
66								0.6	＞10	-	9.0	＞10	≥10	-	25

TABLE 2B (continuation)

Antiviral Activity of control Compounds ^c

Compound (I)	EC50(μg/ml)a							CC50(μg/ml)b
									WT	100-Ile	103-Asn	106-Ala	138-Lys	181-Cys	188-Leu
	67	0.75	≥10	-	≥10	5.5	＞10									-	29
68								0.45	5.0	-	8.5	5.5	8.5	-	＞100
	69	0.04	0.65	-	0.65	0.55	3.5									-	10
70								0.19	2.5	-	6.5	5.5	2.5	-	5.2
	71	0.049	0.6	-	3.0	3.0	1.6									-	4.0
72								0.03	2.3	-	0.5	0.085	1.6	-	24
	73	0.017	2.9	-	0.80	0.09	0.85									-	＞100
74								0.015	6.0	-	0.45	0.09	3.0	-	＞100
	75	0.009	0.9	-	0.7	0.35	0.6									-	16
76								0.035	2.7	-	≥1	0.73	＞1	-	5.3
	77	0.045	≥1	-	≥1	0.55	＞1									-	3.6
78								0.045	5.0	-	2.0	1.13	3.3	-	7.1
	79	0.03	0.85	-	0.07	0.07	0.08									-	4.1
80								0.50	4.0	-	5.0	4.0	3.0	-	8.3
	81	0.04	0.7	-	0.65	0.6	4.5									-	12
82								0.03	2.0	-	1.0	0.55	0.8	-	5.2
	83	0.004	0.17	-	0.057	0.035	0.07									-	9.7
84								0.049	0.6	-	3.0	3.0	1.6	-	4.0
	85	0.5	6.0	-	7.5	5.5	5.3									-	6.7
86								0.1	5.0	-	5.0	4.0	5.0	-	7.4
	87	0.02	3.0	-	1.0	0.55	0.53									-	15
88								0.01	0.7	-	0.6	0.3	0.27	-	5.2
	89	0.09	3.0	-	4.0	1.9	4.0									-	4.9

TABLE 2B (continuation)

Antiviral Activity of control Compounds ^c

Compound (I)	EC50(μg/ml)a							CC50(μg/ml)b
									WT	100-Ile	103-Asn	106-Ala	138-Lys	181-Cys	188-Leu
	90	0.19	3.5	-	6.0	0.7	5.5									-	27
91								0.45	4.5	-	8.5	5.0	7.5	-	5.8
	92	0.02	0.4	-	0.75	0.50	0.37									-	4.3
93								0.03	2.0	-	0.10	0.08	0.7	-	9.7
	94	0.01	0.75	-	0.50	0.15	0.6									-	3.1
95								0.008	0.65	-	0.08	0.16	0.5	-	13
	96	0.05	＞1	-	0.75	0.17	＞1									-	9.0
97								0.009	3.5	-	≥4	2.0	3.3	-	3.3
	98	0.06	＞1	-	0.85	0.7	＞1									-	7.5
99								0.10	＞1	-	＞10	＞10	＞10	-	＞100
	100	0.03	4.0	-	0.8	0.4	0.65									-	＞100
101								0.045	3.0	-	0.2	0.33	0.55	-	≥100
	102	0.03	0.85	-	0.85	0.08	0.65									-	3.7
103								0.01	0.19	-	0.075	0.06	0.07	-	8.5
	104	0.01	0.65	-	0.4	0.2	0.37									-	11
105								0.015	0.4	-	0.2	0.085	0.07	-	6.7
	106	0.007	0.5	-	0.08	0.065	0.085									-	＞100
107								0.04	0.6	-	0.3	0.15	0.3	-	16
	108	0.007	0.27	-	0.08	0.07	0.08									-	7.0
109								0.005	0.17	-	0.04	0.035	0.045	-	4.3
	110	0.04	4.0	-	0.2	0.1	0.5									-	25
111								0.007	0.5	-	0.085	0.077	0.3	-	12
	112	0.025	0.75	-	0.35	0.5	0.8									-	10

TABLE 2B (continuation)

Antiviral Activity of control Compounds ^c

Compound (I)	EC50(μg/ml)a							CC50(μg/ml)b
									WT	100-11e	103-Asn	106-Ala	138-Lys	181-Cys	188-Leu
	113	0.023	0.27	-	0.085	0.15	0.13									-	5.4
114								0.05	0.085	-	0.13	0.075	0.075	-	5.8
	115	0.29	0.65	-	0.2	0.77	0.5									-	4.3
116								0.085	1.8	-	5.5	0.6	2.0	-	≥100
	117	0.06	5.0	-	0.75	0.35	4.0									-	18

^a50% effective concentration (i.e., the concentration of compound required to inhibit 50% of the viral-induced cellular morbidity)^b50% cell static concentration (i.e., the concentration of compound required to inhibit CEM cell proliferation by 50%)^cData are mean values from 2 to 3 independent experiments

Claims (12)

1. A compound of the formula:wherein X is O or S.

2. The compound of claim 1, having the formula:

3. the compound of claim 1, having the formula:

4. a pharmaceutical composition comprising a therapeutically effective amount of a compound of claim 1 and a pharmaceutically acceptable carrier.

5. A pharmaceutical composition comprising a therapeutically effective amount of a compound of claim 2 and a pharmaceutically acceptable carrier.

6. A pharmaceutical composition comprising a therapeutically effective amount of a compound of claim 3 and a pharmaceutically acceptable carrier.

7. Use of a compound according to claim 1 for the manufacture of a medicament for the treatment of HIV-1 infection in an infected host.

8. Use of a compound according to claim 2 for the manufacture of a medicament for the treatment of HIV-1 infection in an infected host.

9. Use of a compound according to claim 3 for the manufacture of a medicament for the treatment of HIV-1 infection in an infected host.

10. The use of a compound according to claim 1 for the preparation of a medicament for inhibiting HIV-1 replication.

11. The use of a compound according to claim 2 for the preparation of a medicament for inhibiting HIV-1 replication.

12. The use of a compound according to claim 3 for the manufacture of a medicament for inhibiting HIV-1 replication.