PL190431B1 - Derivatives of oxazolydinones and pharmacological compositions - Google Patents

Abstract

1. Derivatives of oxazolidinones with formula I: or their pharmaceutically acceptable salts, where: R refers to H or methyl, R1 refers to F, R2 refers to H, R3 refers to methyl, while X refers to O or S.

Description

The invention relates to new derivatives of oxazolidinones or their pharmaceutically acceptable salts, the use of these compounds and a pharmaceutical composition containing these compounds as active substances for the prevention or treatment of infectious diseases. These compounds are the unique oxazolidinones with a hexahydro-1,4-diazepin-5-one substituent.

More specifically, the novel oxazolidinone derivatives of the invention relate to useful antimicrobial agents effective against a variety of human and animal pathogens, including gram positive aerobic organisms such as multi-resistant staphylococci and streptococci, gram negative organisms such as H. influenzae and M. catarrhalis, as well as anaerobic organisms such as bacteroides and clostridia species, and acid resistant organisms such as Mycobacterium tuberculosis and Mycobacterium avium.

International publication WO 97/27188 discloses piperazin-3-one analogs which are homologues of the compounds of the invention.

International publication WO 93/23384 discloses oxazolidinones containing a substituted diazine (piperazine) as part of the molecule, and their use as antibacterial agents.

International publication WO 93/09103 discloses substituted aryls and heteroarylphenyl oxazolidinones useful as antibacterial agents.

International publication WO 90/02744 discloses 5'-indolinyl-5-amidomethyloxazolidinones, 3- (substituted fused ring) tenyl-5-amidomethyloxy) .olidinones, and 3- (substituted nitrogen) phenyl-5-amidomethyloxy) olidinones which are useful as anti-bacterial agents.

Other references disclosing a variety of oxazolidinones include U.S. Patent Nos. 5,547,950, 4,801,600, J Med Chem., 32, 1673-81 (1989); J Med Chem., 33, 2569-78 (1990); Tetrahedron, 45, 1323-26 (1989); and J Med Chem, 35, 1156 (1992).

EP 352 781 discloses phenyl and pyridyl substituted phenyl oxazolidinones.

EP 316 594 discloses 3-substituted styryl oxazolidinones.

EP 312,000 discloses phenylmethyl and pyridylmethyl substituted phenyl oxazolidinones.

Oxazolidinone derivatives according to the invention are represented by the formula I:

or a pharmaceutically acceptable salt thereof, wherein:

R is H or methyl,

R1 is F,

R2 is H,

R3 is methyl and X is O or S.

Preferably, in formula I, X is O.

Preferably, in formula I, X is S.

Preferably, in formula I, R is H.

Preferably, in formula I, R is methyl.

Preferably, in formula I, R3 is methyl.

Preferably, Formula I represents the S enantiomer.

The following compounds are preferred.

(a) (S) -N - [[3- [3-fluoro-4- (1,2,3,4,6,7-hexahydro-5-oxo-1,4-diazepino-1-yl) phenyl ] -2-oxo-5-oxazolidinyl] methyl] acetamide, (b) (S) -N - [[3- [3-fluoro-4- (1,2,3,4,6,7-hexahydro-5 -oxo-1,4-diazepino-1-yl) phenyl] -2-oxo-5-oxazolidine) '] methyl | thioacetamide, (c) (S) -N - [[3- [3-fluoro-4- (1,2,3,4,6,7-hexahydro-4-methyl-5-oxo-1,4-diazepino-1-yl) phenyl] -2-oxo-5-oxazolidinyl] methyl] acetamide, or ( d) (S) -N - [[3- [3-fluoro-4- (1,2,3,4,6,7-hexahydro-4-methyl-5-oxo-1,4-diazepine-1- yl) · phenyl] -2-oxo-5-oxazolidinyl] methyl] thioacetamide.

190 431

The use of oxazolidinone derivatives of formula I as defined in claim 1 1, according to the invention, characterized in that the compounds are used in the manufacture of a medicament for the treatment of microbial infections in humans.

The pharmaceutical composition according to the invention is characterized in that it contains the oxazolidinone derivatives of the formula I as defined in claim 1. 1 and a pharmaceutically acceptable carrier

The antimicrobial agent or pharmaceutical composition comprises an oxazolidinone compound or a pharmaceutically acceptable salt thereof as an active ingredient. An antimicrobial agent containing an active ingredient of the invention may be useful in the treatment or prevention of an infectious disease.

Compounds of Formula I, structurally disclosed above, are useful anti-microbial agents. Typically, as further explained, these compounds may be administered as antibacterial agents at a dose ranging from about 0.1 to 100 mg / kg or preferably from about 3.0 to about 50 mg / kg of body weight per day.

The compounds of the invention can be converted into their salts according to conventional methods.

Pharmaceutically acceptable salts are acid addition salts useful in the administration of the compounds of the invention and include hydrochloride, hydrobromide, sulfate, phosphate, acetate, propionate, lactate, mesylate, maleate, succinate, oxalate, citrate, 2-hydroxyethylsulfonate, fumarate and the like, where the basic group is present. These salts may be in the form of hydrates. Some of the compounds of the present invention can form metallic salts such as sodium, potassium, calcium and magnesium, and these are also encompassed by the term "pharmaceutically acceptable salts".

According to the configuration on the C-5 carbon of the oxazolidinone ring of the compounds represented by the structure of Formula I, the compounds of the present invention may exist in the form of geometric, optical and other isomers, and the invention includes each of these isomers. Both the racemic mixture and the enantiomers are considered useful as antibacterial agents. Regardless, the preferred absolute configuration on the C-5 of the oxazolidinone ring of the compounds is as shown in the structure of Formula I. This absolute configuration is defined as (S) according to the Cahn-Ingold-Prelog nomenclature system. The major part of the pharmacological activity producing the antibacterial effect is believed to be inherent in the (S) enantiomer.

Compounds of formula I can be prepared as shown in Scheme I where P is an alcohol protecting group such as benzyl or ierbutyldimethylsilyl. Structure 2 in this scheme is prepared according to the methods described in example 1, steps 1 and 2. In scheme I, the alcohols of formula 2 are protected as benzyl ethers. In a suitable procedure for this reaction, a solution of alcohol 2 in a solvent such as Et 20 or THF is allowed to react first with sodium hydride at 0-25 ° C and then with benzyl bromide and tetrabutylammonium iodide at 0-25 ° C to obtain structure 3 The ethylene ketal of compound 3 can then be removed with an acid catalyst such as p-toluenesulfonic acid in acetone (as described in Example 1, step 2) to give structure 5 where P is benzyl. Alternatively, the ketal may be removed from structure 2, the resulting structure 4 reacted with tert-butyldimethylsilyl chloride and imidazole in DMF, or with tert-butyldimethylsilyl chloride and triethylamine in methylene chloride, yielding structure 5 in which the alcohol protecting group (P) is tert- butyldimethylsilyl. The ketone 5 is then allowed to react with hydroxylamine hydrochloride and sodium acetate in methanol-methylene chloride to give oxime 6 (see example 1, step 3). The Beckmann rearrangement of structure 6 is carried out with p-toluenesulfonyl chloride and sodium carbonate in aqueous acetone at 20-40 ° C to give structure 7. For compounds of formula I where R is not hydrogen, compounds 7 can be alkylated with R ' Y where Y is Br, I, CH3SO3, or p-CH3 (j> SC> 3 and R 'is a suitable alkyl substituent. In one method of this alkylation, compounds of formula 7 are reacted with sodium hydride and R'Y in a solvent such as DMF at 0-25 ° C to give 8. Alternatively, structure 7 can react with R′Y, potassium hydroxide, and tetrabutylammonium bromide in THF or acetonitrile at 20-50 ° C to give 8 Deprotection of alcohols 7 or 8 provides structure 9. When P is benzyl ether, the reaction can be accomplished by hydrogenolysis with hydrogen and a palladium catalyst in ethanol, or by using ammonium formate and a palladium catalyst in methanol at 10-30 ° C. The tzet-butyl dimethylsilyl protecting group can be removed under acidic conditions or with with a flu ion orcish. Such a deprotection can be carried out, for example, with trifluoroacetic acid in methylene chloride at 25 ° C or with aeteabutylammonium fluoride in THF at 25 ° C to give alcohol 9. The conversion of alcohol 9 to amine 11 can be carried out as described in Example 1, step 1 Alternatively, reaction of 9 with m-nitrobenzenesulfonyl chloride and triethylamine in methylene chloride at 5-25 ° C gives m-nitrobenzenesulfonate 10, which reacts with ammonium hydroxide in THF or acitrile-isopropanol at 30-60 ° C to give amine 11. Reaction. compound 11 with acyl halides, anhydrides, isocyanates, isothiocyanates or dithioesters provides compounds of formula I.

Compounds of formula I where R is hydrogen and X is oxygen are conveniently prepared by allowing compounds 12 to react with p-toluenesulfonyl chloride and sodium carbonate in aqueous acetone at 20-40 ° C (see Example 1, Step 4).

SCHEME

FORMULA I

190 431

The compounds of the present invention are useful for treating microbial infection in humans and other blood-blooded animals by parenteral, oral or topical application.

The term "treatment" as used herein means the partial or total alleviation of symptoms from which the patients suffer; the term "prevention" as used herein means the partial or complete avoidance of disease symptoms in a patient who, as determined by a physician, may be suffering from a disease or related condition, unless countermeasures are taken.

The pharmaceutical compositions of the present invention can be prepared by combining the compounds of formula I of the present invention with solid or liquid pharmaceutically acceptable carriers and, optionally, with pharmaceutically acceptable adjuvants and excipients using standard and conventional techniques. Solid form compositions include powders, tablets, dispersible granules, capsules and suppositories. A solid carrier can be at least one substance, which can also function as a diluent, flavoring agent, solubilizer, lubricant, suspending agent, binder, tablet disintegrating agent and encapsulating agent. Chemically inert solid carriers include magnesium carbonate, magnesium stearate, talc, sugar, lactose, pectin, dextrin, starch, gelatin, cellulosic materials, low melting waxes, cocoa butter and the like. The liquid forms of the compositions include solutions, suspensions, and emulsions. For example, solutions of the compounds of the present invention dissolved in water, water with propylene glycol, or water-polyethylene glycol systems may be provided, optionally containing conventional colorants, flavors, stabilizers, and density enhancing agents.

Preferably, the pharmaceutical composition is prepared by conventional techniques in the form of a dosage unit containing an effective amount of the active ingredient, i.e. a compound of formula I, in accordance with the present invention.

The amount of active ingredient, i. E. A compound of formula I, in the pharmaceutical composition and unit dose of said composition may be varied or adjusted broadly, depending on the particular method of administration, the strength of the particular compound, and the concentration desired. Generally, the amount of active ingredient may vary between 0.5% and 90% by weight of the composition.

In therapeutic use for the treatment or control of bacterial infections in humans and other animals diagnosed with bacterial infections, the compounds or pharmaceutical compositions thereof may be administered orally, parenterally and / or topically at dosages to obtain and maintain a concentration, i.e. , or a blood level of the active ingredient in the treated animal which will be antibacterial. Generally, such antimicrobially effective administered amounts of active ingredient will range from about 0.1 to about 100 mg / kg, more preferably about 3.0 to about 50 mg / kg of body weight per day. It is understood that dosages may vary depending upon the need of the patient, the severity of the bacterial infection being treated, and the particular compound used. It is also understood that the starting dose administered may be increased above the above upper level to rapidly achieve the required blood level, or the starting dose may be less than the optimum, and the daily dose and daily dosage may progressively increase over the course of treatment depending on the particular situation. . If desired, the daily dose can also be divided into multiple doses, for example two to four times daily.

The compounds of formula I are administered parenterally, i.e. by injection, for example by intravenous injection, or by other parenteral administration route. Pharmaceutical compositions for parenteral administration will generally contain a pharmaceutically acceptable amount of a compound of formula I in the form of a soluble salt (acid addition salt or base salt) dissolved in a pharmaceutically acceptable liquid carrier such as, for example, water for injection, and an appropriately buffered isotonic solution, for example having a pH of about 3.5-6. Suitable buffering agents include, for example, trisodium orthophosphate, sodium bicarbonate, sodium citrate, N-methylglucamine, L (+) - lysine, and L (+) - arginine to name a few. The compound of Formula I will generally be dissolved in the carrier

In an amount sufficient to provide a pharmaceutically acceptable injection concentration ranging from about 1 mg / ml to about 400 mg / ml. The resulting liquid pharmaceutical composition is administered so as to achieve the effective dose amount mentioned above. Compounds of formula I of the present invention are preferably orally administered in solid or liquid dosage forms.

For topical application, an effective amount of a compound of Formula I is added to a pharmaceutically acceptable gel or cream base which may be applied to the patient's skin, to the area to be treated. The formulation of such creams and gels is well known to those skilled in the art and may include penetration enhancers.

The compounds of the present invention are useful antimicrobial agents effective against a variety of human and animal pathogens, including gram positive aerobic organisms such as multiply resistant staphylococci and streptococci, gram negative organisms such as H. influenzae and M catarrhalis, as well as anaerobic organisms such as bacteroides and clostridia, and acid resistant organisms such as Mycobacterium tuberculosis and Mycobacterium avium.

In order to more fully illustrate the nature of the invention and the manner of its implementation, the following experimental examples are presented, although they should not be construed as limiting.

Example 1. Preparation of (S) -N - [[3- [3-ίΊuoro-4- (1,2,3,4,6,7-hexahydeo-5-oxo-1,4-diazepine-1-yl) phenyl] -2-oxo-5-oxazolodinyl] methyl] acetamide

Step 1: Preparation of (S) -N- [3- (3-Fluoro-4-piperidin-1-yl-phenyl) -2-oxo-oxazolidine-5-ylmethyl] acetamide

Diisopropylethylamine (15.7 ml) and 3,4-difluoronitrobenzene (5.0 ml) were successively added to a solution of piperidine (5.77 g) in ethyl acetate (70 ml), and the reaction medium was stirred for 2 days at room temperature. Water was added to the reaction solution, and the separated ethyl acetate layer was washed with water and brine, dried over anhydrous sodium sulfate. The solvent was evaporated and the nitro compound (10.1 g) was obtained in 100% yield. Palladium on carbon (10%, 1.0 g) was added to a solution of the nitro compound (10.1 g) in ethyl acetate (101 ml), and the reaction medium was stirred for 4 h. at room temperature under a hydrogen atmosphere. The palladium on carbon was filtered off and the filtrate was concentrated in vacuo to give the amine (8.75 g, 100%). Sodium bicarbonate (5.0 g) and benzyloxythionyl chloride (8.4 ml) were successively added to a tearahydrofuran (THF) solution (100 ml) of the amine (8.75 g) and the reaction medium was stirred for 14 h. in room temperature. Water was added to the reaction solution, and the separated THF layer was washed with water and brine, dried over anhydrous sodium sulfate. The solvent was evaporated and the residue was purified by column chromatography on silica gel (solvent: ethyl acetate / hexane / chloroform = 1/6/4) to obtain benzyl carboxylates (14.5 g) in 98% yield. Butyllithium (1.6 M hexane: 5.2 ml) was added to a solution of benzyl carbamate (2.75 g) in THF (24 ml) at -78 ° C and the reaction medium was stirred for 5 min. At the same temperature, (R) - (-) - glycidyl butyrate (1.25 ml) was added to the stirred solution and the reaction medium was stirred for 14 hours while the temperature slowly rose to room temperature. Water was added to the reaction solution, and the separated THF layer was washed with water and brine, dried over anhydrous sodium sulfate. The solvent was evaporated and the residue was purified by column chromatography on silica gel (solvent: ethyl acetate / hexane = 3/1) to obtain an alcohol (2.20 g) in a yield of 89%.

190 431

Tosyl chloride (2.85 g) was added to a pyridine solution (8 ml) of the alcohol (2.20 g) and the reaction medium was stirred for 6 h. in room temperature. Water (32 ml) was added to the reaction solution, and the reaction medium was stirred 1 hr. The precipitated solid was collected by filtration, washed with water and then dried in vacuo at room temperature to give the tosylate (3.28 g) in a 98% yield. Sodium azide (3.80 g) was added to a dimethylformamide (DMF) solution (23 ml) of tosylate (3.28 g) at room temperature, and the reaction medium was stirred for 5.5 h. at 65 ° C. After the reaction mixture was cooled to room temperature, water was added and the mixture was extracted with ethyl acetate; the organic layer was concentrated in vacuo. The obtained residue was dissolved in ethyl acetate and washed with water and brine, dried over anhydrous sodium sulfate. The solvent was evaporated and the residue was purified by column chromatography on silica gel (solvent: ethyl acetate / hexane = 1/1) to obtain an azide (2.20 g) in a yield of 94%. Acetic anhydride (0.65 ml) and pyridine (1.0 ml) were added to a solution of azide (2.20 g) in ethyl acetate (19 ml) at room temperature, after palladium on carbon (10%, 0.22 g) was added. ), and the reaction medium was stirred for 6 hours at room temperature under 1 atm of hydrogen. Palladium-carbon was filtered off and the filtrate was washed with water and brine, dried over anhydrous sodium sulfate. The solvent was evaporated and the residue was purified by column chromatography on silica gel (solvent: acetone / hexane = 1/1) to obtain the title compound.

Step 2: Preparation of (S) -N- {3- [3-Fluoro-4- (4-oxo-piperidin-1-yl) phenyl] -2-oxo-oxazolidine-5-ylmethyl} -acetamide

Using commercially available 1,4-dioxo-8-aza-spiro [4,5] decane, (S) -N- {3- [4- (1,4-dioxa-8-azaspiro [4.5] decane) was synthesized -8-yl) -3-fluorophenyl] -2-oxo-oxazolidine-5-ylmethyl} -acetamide by the same method as in step 1. Water (3.79 g) was added successively to an acetone solution (70 ml) of this compound (3.79 g) ( 20 ml) and p-toluenesulfonic acid monohydrate (3.66 g) and the reaction medium was heated for 3 h. at boiling point. After cooling the reaction mixture to room temperature, acetone was distilled off and the aqueous layer was neutralized with triethylamine. The solution was extracted with methylene chloride, and the organic layer was washed with brine and dried over anhydrous sodium sulfate. The solvent was evaporated and the residue was purified by column chromatography on silica gel (solvent: chloroform / methanol = 50 / 1-25/1) to give the title compound.

Step 3: Preparation of (S) -N- {3- [3-Fluoro-4- (4-hydroxyiminopiperidin-1-yl) phenyl) -2-oxo-oxazolidine-5-ylmethyl} acetamide

Sodium acetate (517 mg) and hydroxylamine hydrochloride (219 mg) were successively added to a solution of 1.00 g of the product of step 2 in a methanol-methylene chloride mixture (10-10 ml), and the reaction medium was stirred for 2 days at room temperature. The solvent was evaporated and the residue was dissolved in methanol, then silica gel (8 g) was added. Methanol was evaporated and the residue was purified by column chromatography on silica gel (solvent: chloroform / methanol = 50 / 1-25/1) to give the title compound.

Step 4: Preparation of (S) -N - [[3- [3-fluoro-4- (1,2,3,4,6,7-hexahydro-5-oxo-1,4-diazepine-1-yl) phenyl] -2-oxo-5-oxazolidinyl] methyl] acetamide

The mixed reaction medium of compound-product of step 3 (0.200 g, 0.549 mmol) in acetone (5.3 ml), under nitrogen atmosphere, was treated first with 5% aqueous sodium carbonate solution (5.3 ml) and then dropwise over 3 minutes, with a solution of p-toluenesulfonyl chloride (0.16 g, 0.82 mmol) in acetone (2.7 mL). Initially, this mixture is a two-phase solution; however, after about 25 minutes, a precipitate begins to form. The ambient temperature (23 ° C) was maintained for about 4 hours and the mixture was filtered. The filtrate was concentrated under reduced pressure to remove acetone and the aqueous residue was extracted with CH2O2. The extract was dried (MgSO4) and concentrated to give a small amount of crude product. Most of the product is in the aqueous layer which has been concentrated in vacuo. The residue was combined with the crude product from the CH2O2 extract and chromatographed on silica gel with MeOH-NH4OH-CH2Cl2 mixture and continued with 3-5% MeOH and 0.3-0.5% NH4OH. The product was recrystallized from MeOH-EtOAc to afford the title compound. Mp 140-146 ° C;

MS m / z (relative intensity) 364 (M +, 96.1), 320 (100), 306 (6.7), 294 (10.9), 236 (41.8);

HRMS calcd for C17H21FN4O4; 364.1547 (M +); found 364.1545;

190 431 * H NMR [300 MHz, (Cl2 SO] δ 1.81 (s, 3H), 2.57 (m, 2H), 3.07 (m, 4H), 3.24 (m, 2H) , 3.38 (t, 2H), 3.67 (d, d, 1H), 4.06 (t, 1H), 4.68 (m, 1H), 7.08 (t, 1H), 7. 13 (d, d, 1H), 7.45 (d, d, 1H), 7.65 (t, 1H), 8.21 (t, 1H).

Example 2: Preparation of (S) -N - [[3- [3-Fluoro-4- (1,2,3,4,6,7-hexahydro-5-oxo-1,4-diazepine-1-yl ) phenyl] -2-oxo-5-oxazolidinyl] methyl] thioacetamide

C.

It s

^ ch ₃

Step 1: Preparation of (S) - [3- [3-Fluoro-4- (1,2,3,4,6,7-hexahydiO-5-oxo-1,4-diazepine-1-yl) phenyl] ether -2-oxo-5-oxazolidinyl] methyl tert-butyldimethylsilyl

Mixed solution 10.6 g (0.03 mol) (S) - [3- [4- (1,4-dioxo-8-azaspiro [4.5] dec-8-yl) -3-fluorophenyl] -2 -oxo-5-oxazolidinyl] methanol, intermediate of formula 2 (Scheme 1) for the preparation of (S) -N- {3- [3-fluoro-4- (4-oxopiperidin-1-yl) phenyl] -2- oxooxazolidine-5-ylmethyl} -acetamide (Example 1, Step 2) in acetone (230 ml) treated with water (65 ml) and p-toluenesulfonic acid monohydrate (11.4 g, 0.06 mol), heated to reflux at under nitrogen atmosphere for 5 hours and maintained at ambient temperature (24 ° C) for 18 hours. It was then concentrated in vacuo to remove acetone. The aqueous residue was neutralized with sodium bicarbonate and extracted with ethyl acetate; the extract was washed with saturated sodium bicarbonate, water, and dilute sodium chloride, dried (Na 2 SO 4) and concentrated to provide the ketone compound of formula 4 (Scheme 1). A mixed solution of ketone and triethylamine (12.5 mL, 0.09 mol) in methylene chloride (100 mL) was treated with tert-butyldimethylsilyl chloride (6.03 g, 0.04 mol) and kept under nitrogen at ambient temperature for 23 hours. . Additional tert-butyldimethylsilyl chloride (3.0 g) was added and the mixture was kept at ambient temperature for an additional 20 hours. Additional portions of triethylamine (3.0 mL) and tert-butyldimethylsilyl chloride (3.0 g) were again added and the mixture was kept at ambient temperature for 4 days, diluted with methylene chloride, washed with water and dilute sodium chloride solution, dried (NaSO) and concentrated. . Chromatography of the residue on silica gel with acetone-heptane that contained 20-30% acetone gave 7.72 g of zert-butyldimethylsilyl ether (TBDMS), a compound of formula 5 (Scheme 1), where P is TBDMS. A mixed solution of TBDMS ether (7.27 g, 17.2 mmol) in methanol (150 mL) was treated dropwise with a solution of hydroxylamine hydrochloride (1.44 g, 0.021 mol) and sodium acetate (1.72 g, 0.021 mol) in water ( 15 ml) and kept at ambient temperature for 20 hours. The mixture was concentrated under reduced pressure. A solution of the residue, white powder, in methylene chloride, was washed with water and dilute sodium chloride solution, dried (Na2SO4) and concentrated to give 7.25 g of the oxime, a compound of formula 6 (Scheme 1). A mixed solution of oxime in acetone (165 ml), under a nitrogen atmosphere, was treated with a 5% aqueous solution of sodium carbonate (165 ml) followed by a dropwise solution of p-toluenesulfonyl chloride (4.92 g, 0.0258 mol) in acetone (80 ml). The mixture was kept at room temperature for 18 hours and then concentrated under reduced pressure. The methylene chloride solution of the residue was washed with water and dilute sodium chloride solution, dried (Na2SO4) and concentrated. Chromatography of the residue on silica gel with 3% methanol - 0.3% ammonium hydroxide - methylene chloride gave 5.98 g of the title compound.

Step 2: Preparation of (S) - [[3- [3-fluoro-4- (1,2,3,4,6,7-hexahydro-5-oxo-1,4-diazepine-1-yl) phenyl] -2-oxo-5-oxazolidinyl] methyl] imines.

An ice-cold, stirred mixture of the product of Example 2, Step 1 (0.22 g, 0.50 mmol) in tetrahydrofuran (THF; 15 ml), under nitrogen atmosphere, was treated dropwise with

190 431 for 2 minutes with 1M solution of tetrabutylammonium fluoride in THF (1.5 ml). The mixture was kept in an ice bath for 10 minutes and at ambient temperature (24 ° C) for 1 hour 25 minutes, diluted with ethyl acetate, washed with water and brine, dried (NaSO) and concentrated. Chromatography of the residue on silicazel with methanol-methylene chloride containing 3-6% methanol gave 0.15 g of alcohol, a compound of formula 9 (Scheme 1) where R is hydrogen: MS (ES) m / z 324 (M + H +). A stirred suspension of this alcohol (0.15 g, 0.46 mmol) in methylene chloride (15 mL) and THF (8 mL), under nitrogen atmosphere, was treated with triethylamine (0.5 mL, 1.4 mmol) followed by portions of for 1 minute at ambient temperature, m-nitrobenzenesulfonyl chloride (0.14 g, 0.56 mmol). The reaction medium was stirred 90 minutes, mixed with additional methylene chloride (10 ml) to make a solution, and kept at ambient temperature for 1 hour. The medium was then kept at -11 ° C for several days, diluted with methylene chloride, washed with saturated sodium bicarbonate, water and brine, dried (Na2SO4) and concentrated to give 0.21 g of m-nitrobenzenesulfonate, compound of formula 10 (Scheme 1) . The mixed reaction medium of m-nitrobenzenesulfonate (0.21 g, 0.44 mmol), acetonitrile (10 ml), 2-propanol (10 ml) and 29% ammonium hydroxide (10 ml) was heated at 45-50 ° C in a Dry cooler Ice-acetone for 4.5 hours, and kept at ambient temperature for 18 hours. Additional ammonium hydroxide (5 ml) was added and the mixture was heated at 45-50 ° C for 4.5 hours, kept at ambient temperature for 1 hour, treated with 5 ml of ammonium hydroxide and held at ambient temperature for 18 hours. It was then concentrated to give a yellow solid which was chromatographed on silica gel with methanol-methylene chloride containing 5-7.5% methanol followed by 8% methanol-0.2% ammonium hydroxide-methylene chloride to afford the title product.

Step 3: Preparation of (S) -N - [[3- [3-fluoro-4- (1,2,3,4,6,7-hexahydro-5-oxo-1,4-diazepine-1-yl) phenyl] -2-oxo-5-oxazolidinyl] methyl] thioacetamide

A mixed solution of 0.12 g of the product of Example 2, Step 2, and 0.40 ml of triethylamine in a mixture of methylene chloride (10 ml) and methanol (10 ml) under nitrogen atmosphere, was treated with ethyl dithioacetate (0.05 ml) and held at an ambient temperature of 145 hours. Additional 0.05 ml portions of ethyl dithioacetate were added after 24, 31 and 49 hours; additional Methylamine (1.0 ml) was also added after 49 hours. The mixture was concentrated to a small volume, diluted with ethyl acetate, washed with water and brine, dried (Na2SO4 and concentrated. Chromatography of the residue on silica gel with 3.5% methanol-methylene chloride gave 0.061 g of the title product.

Ή NMR [300 MHz, (CDshSO] δ 2.42 (s, 3H), 2.56 (m, 2H), 3.07 (m, 4H), 324 (m, 2H), 3.76 (dd, 1H), 3.87 (m, 2H), 4.11 (t, 1H), 4.91 (m, 1H), 7.12 (m, 2H), 7.46 (dd, 1H), 7, 67 (broad s, EH), 10.35 (broad s, 1H).

Example 3: Preparation of (S) -N - [[3- (3-Fluoro-4- (1,2,3,4,6,7-hexahydro-4-methyl-5-oxo-1,4-diazepine- 1-yl) phenyl) -2-oxo-5-oxazolidinyl) methyl] acetamide

Step 1: Preparation of (S) - [[3- [3-fluoro-4- (1,2,3,4,6,7-hexahydro-4-methyl-5-oxo-1,4-diazepine-1- yl) phenyl] -2-oxo-5-oxazolidinyl] methyl] amine

A mixture of 0.63 g (1.4 mmol) of the product of Example 2, Step 2, methyl iodide (0.093 ml) and THF (40 ml) was added dropwise over 12 minutes, under nitrogen, to a stirred mixture of powdered potassium hydroxide (0 , 12 g) and tetrabutylammonium bromide (0.096 g) in THF (10 ml) and kept at ambient temperature for 20 hours. It was then diluted with ethyl acetate, washed with water and brine, dried (MgSO 4) and concentrated. Chromatography of the residue on silica gel with acetone-methylene chloride, which contained 10-40% acetone, gave 0.46 g (71%) of the methylated product, a compound of formula 8 (Scheme 1) where R 'is methyl. An ice-cold, stirred mixture of this product (0.17 g, 0.38 mmol) and THF (12 mL), under nitrogen atmosphere, was treated dropwise with a 1M solution of tetrabutylammonium fluoride in THF (1.2 mL). It was kept in an ice bath for 15 minutes and at ambient temperature for 3 hours, mixed with ice water and extracted with ethyl acetate. The extract was washed with water and brine, dried (MgSO4) and concentrated to give 0.15 g of the alcohol compound of formula 9 (Scheme 1). A mixed, ice-cold solution of this alcohol (0.52 g,

1.5 mmol) and triethylamine (0.60 ml) in methylene chloride (45 ml) was treated in portions over 5 minutes with m-nitrobenzenesulfonyl chloride (0.42 g). The mixture was kept in an ice bath for 15 minutes and at ambient temperature for 3 hours, diluted with methylene chloride, washed with saturated sodium bicarbonate, water and brine, dried (MgSO 4) and concentrated to give the m-nitrobenzenesulfonate compound of formula 10 (Scheme 1). A mixed mixture of this product, acetonitrile (35 ml), 2-propanol (35 ml) and concentrated ammonium hydroxide (35 ml) was kept at 45-50 ° C in a Dry Ice-acetone cooler for

4.5 hours, and at ambient temperature for 20 hours. Additional ammonium hydroxide (6 mL) was added to the mixture and the mixture was held at 45-50 ° C for

5.5 hours and at ambient temperature for 18 hours. The mixture was then concentrated under reduced pressure to remove the organic solvents and the aqueous residue was extracted first with ethyl acetate and then with methylene chloride. The extracts were washed with water and brine, dried (MgSO4) and concentrated. Chromatography of the residue on silica gel with methanol-methylene chloride containing 7.5-10% methanol gave the title compound.

Step 2: Preparation of (S) -N - [[3- [3-fluoro-4- (1,2,3,4,6,7-hexahydro-4-methyl-5-oxo-1,4-diazepine- l-yl) phenyl] -2-oxo-5-oxazolidinyl] methyl] acetamide

A stirred, ice-cold mixture of 0.10 g (0.30 mmol) of the product of Example 3, Step 1, and pyridine (1.74 mL), under nitrogen atmosphere, was treated dropwise with acetic anhydride (0.57 mL, 6.04 mmol), and kept in an ice bath for 15 minutes and at ambient temperature for 3.5 hours. Then it was concentrated in vacuo; the residue was mixed with ice water and saturated sodium bicarbonate solution and extracted with ethyl acetate. The extract was washed with water and brine, dried (MgSO4) and concentrated. Crystallization of the residue from ethyl acetate-methanol gave 0.053 g of the title compound.

Mp 203-204 ° C.

MS (ES) m / z 379 (M + H +), 401 (M + Na +).

Anal. calcd for C18H23FN4O4: C, 57.13; H, 6.13; N, 14.81.

Found C, 57.05; H, 6.23; N, 14.85.

Example 4. Preparation of (S) -N - [[3- [3-Fluoro-4- (1,2,3,4,6,7-hexahydro-4-methyl-5-oxo-1,4-diazepine- 1-yl) phenyl] -2-oxo-5-oxazolidinyl] methyl] thioacetamide

An ice-cold, mixed solution of 0.18 g (0.535 mmol) of the product of Example 3, Step 1, and triethylamine (0.21 ml) in THF (8 ml) and methylene chloride (10 ml) was treated with a solution of ethyl dithioacetate (0.074 ml, 0.64 mmol) in THF (2 mL). The mixture was kept at 12

Ambient temperature for 20 hours, treated with one additional drop of ethyl dithioacetate and held at ambient temperature for 7 hours. It was then concentrated under a stream of nitrogen. The residue was mixed with methylene chloride, washed with saturated sodium bicarbonate, water and brine, dried (Na ₂ SO 4) and concentrated. Chromatography of the residue on silica gel with methanol-methylene chloride containing 2-4% methanol, and crystallization of the product from ethyl acetate gave 0.13 g of the title compound. Mp 157-158 ° C.

Anal calcd for C1H23FN4O3S: C, 54.81; H, 5.88; N, 14.20.

Found C, 54.83; H, 5.93; N, 14.11.

Example 5 MIC test method

The in vitro MIC values of test compounds were determined by the standard agar dilution method. A drug stock solution for each analog was prepared in the preferred solvent, usually DMSO: H 2 O (1: 3). Serial 2-fold dilutions of each sample were prepared using 1.0 ml aliquots of sterile distilled water. To each 1.0 ml of drug solution was added 9 ml of liquid Mueller Hinton agar medium. Drug-supplemented agar was mixed, poured into 15 x 100 mm petri dishes, and allowed to solidify and dry before inoculating the bacteria into the plates.

Each of the test organism vials was kept frozen in a freezer liquid nitrogen vapor. The test cultures were grown overnight at 35 ° C in an organism-appropriate medium. The colonies were picked with a sterile swab and the cells were suspended in Trypticase Soy broth (TSB) to an turbidity equivalent to 0.5 McFarland standard. 120 dilutions of each of the suspensions were made in TSB. The drug-supplemented agar plates were inoculated with 0.001 ml drop of cell suspension using a Steers replicator, yielding approximately 10 ⁴ to 10 ⁵ cells per spot. The plates were incubated overnight at 35 ° C.

Based on the incubation results, the Minimum Inhibitory Concentration (MIC gg / ml), the lowest drug concentration that inhibits visible organism growth, was read and recorded. The results are presented in Table I.

Table I.

Example No SAUR ^and 9213 MIC SEPI ^b 12084 MIC EFAE ^C 9217 MIC SPNE ^d 9912 MIC HINF ^e 30063 MIC MCATf 30610 MIC 1 4 1 4 0.5 8 8 3 4 1 4 1 32 8 4 1 <0.5 1 <0.5 8 2

a) S aureus, culture 9213

b) S. epidermidis, culture 12084

c) E faecalis, culture 9217

d) S pueumoniae, culture 9912

e) H influenzae, culture 30063

f) M catarrhalis, culture 30610

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Claims (10)

Patent claims 1. Oxazolidinone derivatives of formula I: FT 'and N, or a pharmaceutically acceptable salt thereof, wherein: R is H or methyl, Ri is F, R2 is H, R3 is methyl and X is O or S. 2. The compound according to claim l and characterized by that zz in formula I X is O. Compound according to claim 3 li with this, with in the cavity I X about S. 4. The compound of claim 1 1, for example, in that IR is H. 5. The compound according to p. and characterized by Zym, Z in formula I R is methyl. 6. The compound according to p. 1, except that IR 3 in the formula is methyl. 7. The compound of claim 1 1, naaminaay in that formula I represents the S enantiomer. A compound according to claim 8 1, it is based on the fact that it is (a) (Si-N-fp - ^ - ffuoroU-fOJNY UdiexahydiO-S-oxo-lN-diazepine-l-yloffenYloyU-oxo-5-oxszolidinyl] methyl] sitamide, (b) ( S) -N - [[3- [3-fluoro-4- (1,2,3,4,6,7-hexahydro-5-oxo-1,4-diazepine-1-yl) phenyl] -2- oxo-5-oxazolidinyl] methyl] thioacItsmid, (c) (S) -N - [[3- [3-fluoro-4- (1,2,3,4,6,7-hexahydro-4-methyl-5) -oxo-1,4-diazIpino-1-yl) phenyl] -2-oxo-5-oxazolidinyl] methyl] acetamide, or (d) (S) -N - [[3- [3-fluoro-4- ( 1,2,3,4,6,7-hexahydro-4-methyl-5-oxo-1,4-diazIpino-1-yl) phenyl] -2-oxo-5-oxycholidinyl] methyl] thioacItamide. 9. The use of the oxazolidinone derivatives of formula I as defined in claim 1 1 for the manufacture of a medicament for the treatment of microbial infections in humans. 10. A pharmaceutical composition containing the oxazolidinone derivatives of the formula I as defined in claim 1. 1 and a pharmaceutically acceptable carrier.