HU181457B - Process for producing azabicycloheptane derivatives - Google Patents

Description

Process for the preparation of azabicycloheptene derivatives

The present invention relates to a novel antibiotic 7-oxo-1-azabicyclo [3.2.0] hept-2-ene-2-carboxylic acid derivative of formula I wherein R ⁶ is hydroxy (1-4 carbon atoms). containing a) alkyl,

R ^{7 is} hydrogen or methyl,

R ^B is C 1-6 alkyl optionally substituted with hydroxy, carboxy, benzyloxycarbonyl or C 2 -C 5 alkoxycarbonyl, unsubstituted pyridyl, or - (CH ₂ ) _n -NQT, and in the latter

T is hydrogen or -C (NH) -NH ₂ or -CH = NH,

Q is hydrogen or C 1-4 alkyl, n is an integer from 1 to 8, and the alkylene chain may be optionally substituted with carboxyl, provided that when R ^{7 is} hydrogen and R ⁶ is 2-hydroxyethyl, then R ⁸ is other than 2-aminoethyl or an N-substituted derivative thereof for the preparation of their pharmaceutically acceptable salts.

U.S. Patent No. 3,950,537 discloses thienamycin, an antibiotic structure analogous to that of Formula I, i.e. 3- (2-aminoethylthio) -6- (1-hydroxyethyl) -7-oxo. l-azabicyclo [3.2.0] hept-2-ene-2-carboxylic acid.

Extremely important szű181457 narrower group of the compounds of Formula I consists of compounds of Formula Ia wherein R ⁸ is as defined above.

It is well known to those skilled in the art that there is a constant need for doctors for new antibiotics. This is due to the unfortunate fact that any given antibiotic does not have constant efficacy because its continuous widespread use results in the development of resistant strains of that pathogen. A further disadvantage of the known antibiotics is that they are effective only against certain types of microorganisms. Therefore, research into new antibiotics is continuing throughout the world.

Thus, it has been found that compounds of formula I are a new class of antibiotics that are useful in both human and veterinary medicine and inanimate systems as they are broadly effective against pathogens such as Gram-positive bacteria (e.g., Staphylococcus). auresus, Streptomyces pyogenes and Bacillus subtilis) and Gram-negative bacteria (e.g., Escherichia coli, Proteus morganii,

Serratia, Pseudomonas and Klebsiella bacteria).

Among the compounds of formula I, the compounds of formula Ia can be prepared by total synthesis as depicted in Scheme A.

The synthetic synthesis depicted in Scheme A is carried out as described below.

The starting material 4- 4- (2-substituted-vinyl-2-azetidinone) is prepared by reacting 1 R'-oxybutadiene with 2 chlorosulfonyl isocyanate.

The reaction of -1181457 is optionally carried out in the presence of a solvent such as diethyl ether, ethyl acetate, chloroform or methylene chloride at a temperature of -78 ° C to + 25 ° C for a few minutes to 1 hour to give compound 3. The R * group is an easily removable acyl-type protecting group (for example, an alkanoyl or aralkanoyl group) that contains a group or groups which do not interfere with the reaction sequence to be carried out. (1 + 2 3 4). Intermediate 3 is first converted to the corresponding sulfinamide derivative by reduction and then hydrolyzed to pH 6-8 to give the corresponding compound 4. Typically, the reaction mixture containing Compound 3 is contacted with an aqueous solution of a reducing agent such as sodium sulfite or thiophenol at 0 to 25 ° C and pH 6-8 for 5 to 30 minutes to afford Compound 4.

Reaction 4 to 5 is accomplished by reduction, and preferably by hydrogenation, such that the compound to be reduced is in a solvent such as diethyl ether, ethyl acetate, dioxane, tetrahydrofuran or ethanol at a temperature between 0 ° C and 25 ° C for 5 minutes to 2 hours. for 1 to 10 atm. in the presence of a hydrogenation catalyst such as platinum, platinum oxide or 10% palladium on carbon.

The reaction of 5-6, i.e. deprotection, is usually conveniently carried out when R ¹ is acyl. The purpose of the deprotection is to allow the subsequent alkylation step, i.e. the 7-to-8 reaction. The deprotection is preferably carried out by alcoholysis in a C 1 -C 6 alkanol such as methanol or ethanol in the presence of the corresponding alkali metal alcoholate such as sodium methylate. The reaction time is usually 5 minutes to 1 hour and the reaction temperature is -10 ° C to + 25 ° C.

The 6-7 reaction is then carried out, i.e., the protecting groups R ² and R ³ are introduced into compound 6 in order to obtain a suitably protected compound for the subsequent alkylation step. Any protecting groups may be utilized for this purpose provided, of course, that they do not inhibit the alkylation desired. R ³ can be hydrogen, a triorganosilyl group such as trimethylsilyl group, or a cyclic ether group such as 2-tetrahydropyranyl group. R ² can be a cyclic ether group such as 2-tetrahydropyranyl. Alternatively, R ² and R ³ may be joined together to form a protected compound 7a. The latter compound can be readily prepared in the presence of Compound 6 and a 2,2-dimethoxypropane catalyst such as boron trifluoride diethyl ether complex or toluenesulfonic acid at a temperature of -10 ° C to + 35 ° C for a few minutes to 1 hour, e.g. by reaction with methylene chloride, diethyl ether, chloroform or dioxane.

The alkylation reaction> 8 is preferably carried out by treating the compound 7 with a strong base such as lithium diisopropylamide, sodium amide or potassium hydride in a solvent such as tetrahydrofuran, ethylene glycol methyl ether, diethyl ether, dimethylformamide or dimethylformamide. sulfoxide at -78 ° C to 0 ° C followed by treatment of the resulting anion with excess acetaldehyde to give Compound 8.

In the reaction AS -> 9, the protecting group R ^{4 is} applied and is usually carried out by treating the compound 8 with a base such as an alkali metal hydroxide, lithium diisopropylamide, 4-dimethylaminopyridine or n-butyllithium in a solvent, e.g. diethyl ether, tetrahydrofuran 2 ban, dioxane, dimethylformamide or dimethylsulfoxide and then for 1 to 24 hours with an acyl halide such as an alkanoyl, aralkanoyl or ring-substituted aralkanoyl halide, or optionally an alkyl halogen with a substituted aryl or optionally substituted aralkyl ester such as 4-nitrobenzyl chloroformate.

The deprotection reaction 9 to 10 is usually carried out by acid hydrolysis, for example by treatment with aqueous acetic acid at a temperature between 25 ° C and 75 ° C for 5 minutes to 3 hours.

The aldehyde compound 77 is prepared by reacting compound 10 with an oxidizing agent such as chromium (IV) oxide-dipyridine complex in cyanomethane, a 1: 1 mixture of dimethylsulfoxide and acetic anhydride, or cyclohexylcarbodiimide in dimethyl sulfoxide. at 5 minutes to 1 hour. The resulting compound 77 is then treated with an excess of HSR ⁸ , where R ⁸ is as defined above, in a solvent such as acetonitrile, methylene chloride, or chloroform, where R ⁸ is as defined above, in an acidic catalyst such as boron trifluoride. - diethyl ether in the presence of complex or toluenesulfonic acid. The reaction usually requires 1-60 minutes.

The vinyl sulfide derivative 14 is obtained via intermediate 13 by treating compound 72 with a halogen such as chlorine or bromine (X = chlorine or bromine) in a solvent such as diethyl ether, methylene chloride, tetrahydrofuran or ethylene glycol methyl ether at -78 ° C and + 30 ° C. Intermediate 13 with an olefin such as cyclohexane or isobutylene is treated with a base such as triethylamine, 1,5-diazabicyclo [5.4.0] undec-5-ene at a temperature of 1 to 30 minutes and immediately thereafter. hereinafter referred to as "DBU") or in the presence of sodium hydride treated with a solvent such as dimethylformamide, ethylene glycol methyl ether, tetrahydrofuran or hexamethylphosphoric acid amide for 1 to 8 hours at a temperature between -20 ° C and + 25 ° C.

The vinyl sulfide derivative 14 is then reacted with the di-ester of oxomalonic acid (or its monohydrate) to afford compound 75. The oxomalonsav ^R5 substitute, ie, esters of the parameter is not critical, e.g., R ^e may be any conventional, readily removable protecting group or a pharmaceutically acceptable ester-forming group. R ⁵ report examples thereof include 4-nitrobenzyl, benzyl, 2-nitrobenzyl, t-butyl and 2,2,2-trichloroethyl group. The reaction 14 to 15 is usually carried out in a high boiling organic solvent such as benzene, toluene, cyclohexane or a halogenated aromatic solvent at a temperature between 50 ° C and the reflux temperature of the reaction mixture for 0.5 to 6 hours.

The 15 to 16 halogenation reaction is usually carried out in a solvent such as tetrahydrofuran, ethylene glycol methyl ether, methylene chloride or chloroform with a halogenating agent such as thionyl chloride or phosphorus pentachloride in the presence of a base such as pyridine at -20 ° C to + 25 ° C. at a temperature of 5 minutes to 3 hours. The selective reduction 15 through 17 of intermediate 16 is terminated by treating compound 16 with, for example, tributylphosphine or triphenylphosphine in a mixture of water and dimethylformamide or a similar aqueous medium (such as dioxane, tetrahydrofuran, ethylene glycol, sulfoxide or acetone) at 0-50 ° C for 10 minutes to 5 hours.

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Compound A7 is then halogenated as described in Reaction 12-13, except that the addition of cyclohexene or other olefin is omitted. The resulting dihalogenated compound 18 is then treated with a base such as triethylamine, sodium hydride or potassium hydride in a solvent such as dimethylformamide, acetonitrile, methylene chloride, chloroform or ethylene glycol methyl ether at -78 ° C and + 25 ° C. between 1-5 hours. Compound 19 is then converted to Compound 20 with a strong base such as DBU or 1,5-diazabicyclo [3.4.0] non-5-ene (hereinafter abbreviated as DBN) in a solvent such as dimethylsulfoxide, acetone, chloroform. , dimethylformamide, tetrahydrofuran or ethylene glycol methyl ether, or silver fluoride in pyridine by treatment at 0-40 ° C for 0.25 to 24 hours. For the reaction 20 to 21, compound 20 is treated with an aromatic base such as pyridine, S-collidine or lutidine or aqueous dimethylsulfoxide in the presence of a displacement agent such as lithium iodide, sodium chloride or sodium bromide. 80-150 ° C for 15 minutes to 2 hours. Treatment of the reaction mixture with aqueous medium yields 21. The double bond isomerization, i.e. reaction 21 to 22, is carried out by treating 21 with a strong base such as diisopropylamine, DBU or DBN in a solvent such as dimethylformamide, dimethyl sulfoxide, diethyl ether. , tetrahydrofuran, ethylene glycol methyl ether or methylene chloride at 0-25 ° C for a few minutes to 2 hours or until equilibrium is reached. (The latter can be determined by ultraviolet absorption assay or by thin layer chromatography.) The final reaction 22-1a, i.e. hydrogenolysis of the protecting groups, is carried out by reacting 22 in a solvent such as dioxane, ethanol or tetrahydrofuran, or in the presence of palladium on carbon, treated with hydrogen at a pressure of 1-4 atm for 0.5-8 hours at 0-25 ° C.

In carrying out the above-described total synthesis, instead of the 4 4- (2-substituted-vinyl) -2-azetidione derivatives, it is also possible to start from the 23 4-vinyl-2-azetidione (prepared by Moriconi, E.J. and Meyer, W.C. J. Org. Chem., 36, 2841 (1971)]. The advantage of this variant of total synthesis is that stereoselective synthesis can be achieved early in the process. Scheme B thus illustrates this variant of the synthesis that is attached to compound 14 to the total synthesis depicted in Scheme A.

The reactions shown in Scheme B are described in more detail below.

Compound 23 is first silylated to provide the N-silylated derivative 24. In the parent group, R 'is an alkyl group containing from 1 to 6 carbon atoms. Trimethylsilyl or tert-butyldimethylsilyl are particularly preferred. The silylation reaction 23 to 24 is typically carried out by reacting 23 in a solvent such as dimethylformamide, dimethylsulfoxide or hexamethylphosphoric acid with a suitable silylating agent such as tert-butyldimethylsilyl chloride and a base, for example, triethylamine, pyridine or N, N-dimethylaniline at a temperature of -10 ° C to + 30 ° C for 1-8 hours. Compound 24 is then alkylated to form compound 25 by treatment with acetaldehyde in the presence of a base. Reactions 24 to 25 are carried out in exactly the same manner as reactions 8 to 9 described above. The O-protecting group (i.e., the oxygen protecting group) a

-> 26 reactions. The protecting group R ^{4 has the} meaning given above and thus reaction 25 to 26 is exactly analogous to the previous reaction 8 9. It is noted here that the reactions 24 to 25 and 25 to 26 provide a good opportunity to separate the compounds 25 or 26 to their racemic diastereomers, if desired. The cleavage of the N-silyl group is carried out in reaction 26-27 by weak acid catalyzed solvolysis. From Compound 27, Compound 28 is prepared by reacting the above solvent in a solvent such as methylene 10 -chloride, tetrahydrofuran or ethylene glycol-methyl ether with an reagent of the formula XSR ⁸ wherein R ⁸ is as previously defined and X is a halogen atom such as chlorine or bromine. Reacting at -50 ° C to + 50 ° C for 1-16 hours. From compound 28, compound 14, which is utilized in the total synthesis depicted in Scheme A, is obtained by removing one HX molecule from compound 28 with a strong base such as DBU, DBN, 1,4-diazabicyclo [2.2.2] octane (hereinafter abbreviated). DABCO) or silver fluoride in a solvent such as dimethylformamide, dimethylsulfoxide, pyridine or hexamethylphosphoric acid amide at -20 ° C to + 50 ° C for 0.25 to 16 hours.

The compounds of formula I form pharmaceutically acceptable salts with inorganic and organic bases. Thus, for example, alkali metal and alkaline earth metal, hydroxide, carbonate and bicarbonate salts, as well as primary, secondary and tertiary amines such as monoalkyl, dialkyl and trialkylamines, have from 1 to 4 carbon atoms in their alkyl form. containing alkanol or dialkanolamines, alkylenediamines, N, N-di-aralkylalkylenediamines, aralkylamines, amino-substituted alkanols, N, N-dialkylamino-substituted alkanols, amino-, polyamino- and guanidino-substituted alkanoic acids, and nitrogen-containing heterocyclic amines can be used. Typical bases of the above types include sodium hydroxide, sodium carbonate, sodium bicarbonate, potassium carbonate, potassium hydroxide, calcium carbonate, trimethylamine, triethylamine, lysine, protamine, arginine, procaine, ethanolamine, morpholine, benzylamine, ethylenediamine, Ν, Ν'-dibenzylethylenediamine, diethanolamine, piperazine, dimethylaminoethanol, 2-amino-2-methyl-1-propanes , theophylline and N-methylglucamine. Pharmaceutically acceptable acid addition salts may also be prepared, for example, by reaction with hydrochloric acid, tartaric acid, hydrobromic acid, sulfuric acid, nitric acid, p-toluenesulfonic acid or methanesulfonic acid.

The salts may be mono-salts, for example, the monosodium salt may be obtained by reacting one molar equivalent of the compound of formula I with one molar equivalent of sodium hydroxide, or may form mixed salts. The latter may be prepared by reacting a molar equivalent of a divalent cation base, such as calcium hydroxide, with a molar equivalent of a compound of formula (I). Pharmaceutically acceptable salts of the compounds of Formula I may be used as active ingredients in the form of suitable pharmaceutical compositions or may be used in combination with other active ingredients to widen the spectrum of action.

Thus, the compounds of the present invention are valuable antimicrobial agents that are effective against both Gram-positive and Gram-negative pathogenic microorganisms. Thus, the free acids or free bases of the formula I, in particular their salts,

Their salts and metal salts, preferably the alkali metal and alkaline earth metal salts, are effective bactericides and can be used, inter alia, to remove pathogenic pathogens from dental and medical equipment, to isolate microorganisms and for human and veterinary use. For the latter, pharmaceutically acceptable salts of inorganic and organic bases, such as salts known from the chemistry of cephem carboxylic acids and penicarboxylic acids, are utilized. For this purpose, for example, alkali metal and alkaline earth metal salts as well as salts with primary, secondary and tertiary amines can be utilized. These salts can be formulated with pharmaceutically acceptable, liquid and solid carriers and / or excipients into standard pharmaceutical preparations such as pills, tablets, capsules, suppositories, syrups or elixirs. 15

The compounds of the present invention are useful in human and veterinary medicine for their activity against Gram-positive and Gram-negative bacteria, such as Staphylococcus aureus, Escherichia coli, Klebsiella pneumoniae, Bacillus subtilis, Salmonella typhosa, Pseudomonas, and Bacteria. to treat infections.

The compounds of the present invention can also be used as additives in animal nutrition, for protecting foodstuffs from bacterial infections and as a disinfectant. For example, they may be used in aqueous formulations in amounts of 0.1 to 100 parts per million of aqueous solution to inhibit or kill the growth of pathogenic bacteria, and as industrial bactericides, such as aqueous-based paints and papermaking inhibiting the growth of bacteria.

One or more of the compounds of the invention may be used in any type of pharmaceutical preparation. Capsules, tablets, powders and liquid solutions, suspensions, or elixirs may conveniently be used as such pharmaceutical compositions. These compositions may be administered orally, intravenously or intramuscularly.

Most preferably, the pharmaceutical compositions are in such a form that they can be absorbed in the gastrointestinal tract. Tablets and capsules for oral administration are preferably in unit dosage form and contain conventional carriers and / or excipients, such as excipients (e.g., sugar syrup, gelatin, acacia, sorbitol, tragacanth or polyvinylpyrrolidine), fillers (e.g., lactose, corn starch, calcium phosphate, sorbitol or glycine), lubricants (e.g., magnesium stearate, talc, polyethylene glycol or silica), disintegrants (e.g., potato starch) or pharmaceutically acceptable ). The tablets may be coated with conventional coatings. Liquid preparations for oral administration may also be aqueous or oily suspensions, solutions, emulsions, syrups or elixirs, or may be prepared in the dry state. The latter form must be reconstituted with water or other suitable liquid carrier prior to use.

Liquid formulations may also contain conventional additives such as 6C suspending agents (e.g., sorbitol syrup, methylcellulose, glucose sugar syrup, gelatin, hydroxyethylcellulose, carboxymethylcellulose, aluminum stearate gel or hydrogenated digestible oils, almond oil, fractionated coconut oil, oily esters, propylene glycol or ethanol) and preservatives, for example 65, a mixture of 4-hydroxybenzoic acid methyl ester and 4-hydroxybenzoic acid propyl ester or sorbic acid. Suppositories may contain a conventional suppository base such as cocoa butter or other glyceride.

Formulations for injection may be presented in unit dose form in ampoules or in multi-dose containers using a preservative. These compositions may be in the form of suspensions, solutions, or emulsions in oily or aqueous vehicles, and may contain formulating agents such as suspending, stabilizing and / or dispersing agents. Alternatively, the active ingredient may be in powder form and should be reconstituted with a suitable vehicle, e.g., sterile, antipyretic, water prior to use.

The pharmaceutical compositions may also be formulated so as to be capable of being absorbed by the mucous membranes of the nose and throat or by bronchial tissue. For this purpose, these pharmaceutical compositions may be formulated, for example, in the form of a powder, liquid spray, inhaler, lozenge or throat. For the treatment of the eye or ear, the compositions may be presented as individual capsules, in liquid or semisolid form, or in the form of drops. Formulations suitable for topical administration with hydrophobic or hydrophilic ingredients include ointments, creams, lotions, powders, toppings, and the like. we can make it.

As mentioned above, pharmaceutical compositions may contain, in addition to carriers, other additives such as stabilizers, binders, antioxidants, preservatives, lubricants, suspending agents, viscosity regulating agents, or flavoring agents. In some cases, other active ingredient (s) may be incorporated into the compositions to widen the spectrum of action.

For veterinary use, the composition is formulated, for example, as an intramammary composition using slow or rapid release ingredients.

The amount of dose to be administered will be highly dependent on the condition and weight of the subject being treated, the mode and frequency of administration. For general infections, parenteral administration is preferred, while for visceral infections, oral administration is preferred. Generally, the daily oral dose is about 5 to 600 mg of active ingredient per kilogram of body weight, administered once or several times a day. A preferred daily dose range for an adult human is about 15 to 240 mg / kg body weight.

Thus, the pharmaceutical compositions of the present invention can be used in a variety of dosage units, for example, solid or liquid, for oral administration. The compositions contain from 1 to 99% by weight of the active ingredient, preferably about 10 to 60% by weight, or about 15 to 1500 mg of active ingredient per unit dose, whether solid or liquid. Of the latter concentration range, 250 mg to 1000 mg are preferred. For parenteral administration, the dosage unit will usually be presented as a solution of the pure active ingredient in slightly acidic sterile water or as a soluble powder.

The following examples illustrate the invention.

Example 1

Preparation of 4- (2-Acetoxyvinyl) -2-azetidinone (Scheme C)

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Under a nitrogen atmosphere, a solution of 1.0 ml of distilled chlorosulfonyl isocyanate (1.65 g, 11.7 mmol) in 2.5 ml of anhydrous diethyl ether was cooled in a bath at -20 ° C, followed by the addition of 2.5 g (22 mmol) of 1-acetoxy- of butadiene in 2.5 ml of anhydrous diethyl ether and the like cooled dropwise using a Teflon tube dipped in chlorosulfonyl isocyanate solution and pressurized with nitrogen gas. Dosing takes 10 minutes. No or little color change. The reaction mixture was then stirred at -20 ° C for 0.5 h. The solution thus obtained is clear, turbid and pale yellow in color.

During the aforementioned 0.5 hour stirring period, a solution of 2 g of sodium sulfite and 5 g of potassium hydrogen phosphate in 20 ml of water was prepared and cooled in an ice bath. To the solution was added diethyl ether (20 mL) and the resulting mixture was vigorously stirred while cooling in an ice bath. Following the aforementioned 0.5 hour stirring period, the reaction mixture is again passed from a reaction vessel maintained in a bath at -20 ° C, using Teflon tube and nitrogen gas, to a vigorously stirred hydrolysis mixture. The rapid dropwise addition is performed within 5 minutes. The reaction mixture was allowed to stand for a further 5 minutes to ensure completion of the hydrolysis. The pH of the reaction mixture is 6-8, preferably 8.

The phases are then separated, leaving the yellow-orange resin in the aqueous phase. The diethyl ether phase was dried directly with magnesium sulfate. The aqueous resin phase is extracted three times with 50 ml of diethyl ether, each time the extracts are added to the organic phase containing magnesium sulfate.

The dried organic phase was then filtered and concentrated under a stream of nitrogen to 5 ml; part of the product crystallizes out at this stage.

A column of g Baker silica gel (stored in diethyl ether) is prepared and the diethyl ether concentrate obtained as described in the previous paragraph is applied to the column and allowed to flow. The flask containing the concentrate was rinsed three times with 2 ml of diethyl ether, pipetted with each rinse and applied to the column. Elution with diethyl ether is then started. The first 25 ml is primarily a gap volume. Five 10 to 10 ml fractions were separated five times and then 50 to 50 ml fractions three times, each concentrated under a stream of nitrogen. 4-6. The product crystallizes from fractions 1 to 3, fractions 3 and 7 only in traces. 1-3. The fractions contain a yellow, pungent odor which is waxy on standing. Yield: 100 mg as a mixture of the cis and trans isomers.

Example 2

Preparation of 4- (2-Acetoxyethyl) -2-azetidione (Scheme D) 10.0 g (0.065 mol) in a Parr hydrogenator

A solution of 4- (2-acetoxyvinyl) -2-azetidinone in ethyl acetate (200 mL) containing palladium on carbon (100 mg, 10% metal) was hydrogenated at 20 ° C with 2.1 atm of hydrogen for 15 minutes. filtered through a bed of filter material and the latter washed with additional ethyl acetate. The combined filtrate was concentrated in vacuo to give the title compound as a crystalline solid (10.0 g). Recrystallization from diethyl ether gives crystals, m.p. 44-47 ° C.

IR spectrum (CHCl ₃ ): δ 5.66 and 5.74.

NMR (CDC1 _3): τ 3.44 (br s, 1, NH), 5.82 (m, 2, CH ₂ OCOCH _3), 6.29 (m, 1, C ^ H), 6.87 [ 'Λ B band into four further cleaved C-4H and NH result, 1, J _gem = 12.8 Hz, J = 4.5 H J _NH = 1.9 Hz], 7.38 ^[l Á Band B is further split into four portions by C-4H and NH, 1, J = 12.8 Hz, J = 2.3 Hz, J = 1.0 Hz], 7.93 and 8.02 (sm , heron 'NH ^{J v} total 5, OCOCtf ₃ and CH ₂ CH ₂ OCOCH ₃ ).

Example 3

Preparation of 4- (2-Hydroxyethyl) -2-azetidinone (Scheme E)

To a solution of 2.24 g (0.014 mol) of 4- (2-acetoxyethyl) -2-azetidinone in 25 ml of anhydrous methanol in a nitrogen atmosphere at 0 ° C was added a solution of 77 mg (1.4 mmol) of sodium methylate in 5 ml of anhydrous methanol. After stirring for 1 hour, the reaction mixture was neutralized by the addition of glacial acetic acid. The methanol is then evaporated in vacuo to give the crude title compound as an oil. The crude product is then purified by chromatography on silica gel using 10% methanol in chloroform as eluent. 1.55 g of the title compound are obtained, m.p.

IR spectrum (CHCl ₃ ): µ 5.67.

Nuclear Magnetic Resonance Spectrum (CDCl ₃ ) δ 3.20 (bs, 1, NH), 6.24 and 6.28 (m in t, total 3, C-4H and C77 ₂ OH), 6.90 (broad s in the 1 '/ lAB band which was split into another four by C-4H and NH, totaling 2, OH and C-3H, J = 13.0 Hz, J = 4.2 Hz, J = 1.6 Hz), 7.42 (Vi AB band boom vic NH ^K further split into C-1H and NH, 1, C-3H, J = 13.0 Hz, J = 2.2 Hz, J = 1.1 Hz) and 8.16 (m, 2, C77 ₂ CH ₂ OH).

Example 4

8-Oxo-2,2-dimethyl-3-oxa! Preparation of azabicyclo (4.2.0) octane (Scheme F)

A solution of 4- (2-hydroxyethyl) -2-azetidione (1.87 g, 0.016 mol) and 2,2-dimethoxypropane (1.69 g, 0.016 mol) in anhydrous methylene chloride (25 ml) was added at 25 ° C. Boron trifluoride-diethyl ether complex (0.201 ml, 0.002 mol) was added and the resulting mixture was stirred for 10 minutes and the solvent was evaporated under reduced pressure to give 2.5 g of an oil which was purified by chromatography on silica gel with ethyl acetate and benzene. 2: 1 to give the title compound (1.59 g) as a crystalline solid, m.p. 60-61 ° C after recrystallization from diethyl ether / hexane.

IR (CHC1 _3): 5.73 μ (beta-lactam)

Nuclear Magnetic Resonance Spectrum (CDCl ₃ ) δ 6.02-6.28 (m, 2H, C-4 methylene), 6.22-6.62 (m, 1H, C-6 methine), 6.90 (dd) , 1H, J _?? = 14 Hz, J ₆₇ = 4.5 Hz, C-7 proton cis - C-6H), 7.47 (dd, 1H, J = 14 _{7, 6} J = _2,? C-7 proton trans - C-6H), 7.82-8.68 (m, 2H, C-5 methylene), 8.23 (s, 3H, C-2 methyl) and 8.57 (s, 3H C-2 methyl)

Example 5

Preparation of 8-Oxo-2,2-dimethyl-7α and -b- (1-hydroxyethyl) -3-oxa-1-azabicyclo (4.2.0) octane (Scheme G)

1.1 molar equivalents at -78 ° C under nitrogen

-5181457 To a solution of freshly prepared lithium diisopropylamide in anhydrous tetrahydrofuran, 8-oxo-2,2-dimethyl-3-oxa-1-azabicyclo (4.2.0) octane was prepared with anhydrous tetrahydrofuran and previously cooled to -78 ° C. solution. After 2 minutes, the lithium enolate formed was treated with excess acetaldehyde, and the reaction mixture was stirred for 30 minutes at -78 ° C and then poured into water. The aqueous phase is then saturated with sodium chloride and extracted with ethyl acetate. The ethyl acetate layer was dried over magnesium sulfate, filtered, and the filtrate evaporated under reduced pressure. The crude product thus obtained was purified by chromatography on silica gel using ethyl acetate / benzene as eluent. This gives the title product mixture. The identity of 8-oxo-2,2-dimethyl-7β- (1-hydroxyethyl) -3-oxa-1-azabicyclo (4.2.0) octane is as follows:

IR spectrum (CH ₂ Cl ₂ ): µ 5.72 (beta-lactam ring). Nuclear Magnetic Resonance Spectrum (CDCl ₃ ): τ 5.53-6.43 (η, 4H, C-4 methylene + C-6 methine + C-5 methine), 6.90 (dd broad s, 2H, J ₇₉ = 9 Hz, J _6? = 5.5Hz, C-7 methine + OH), 7.70 to 8.83 (m, 2H, C-5 CH), 8.27 (s, 3H, C-2 methyl ), 8.60 (s, 3H, C-2 methyl) and 8.78 (d, 3H, J ₉₁₀ = 6.5Hz, C-10 methyl). The identity of 8-oxo-2,2-dimethyl-7a- (1-hydroxyethyl) -3-oxa-1-azabicyclo (4.2.0) octane is as follows:

IR spectrum (CHCl 3): µ 2.9 (broad O-H), 5.73 (beta-lactam ring)

Nuclear Magnetic Resonance Spectrum (acetone-D _e ): τ 3.33-4.23 (m, C-9 methine + C-4 methylene + C-6 methine), 3.33 (bs, OH), 2, 83 (dd, J = 2 Hz, 6 Hz _χ, C-7 methine), 2.67 (dd, J = 2 Hz, 8 Hz, C-7 methine), 1.63-1.93 (m, C -5 methylene), 1.63 (s, C-2 methyl), 1.40 (s, C-2 methyl) and 1.23 (d, J = 6.5 Hz, C-10 methyl).

Example 6

Preparation of 8-Oxo-2,2-dimethyl-7α- [1- (4-nitrobenzylcarbonyldioxy) ethyl] -3-oxa-1-azabicyclo (4.2.0) octane (Scheme H) on anhydrous conditions 60 mg (0.302 mmol) of 8-oxo-2,2-dimethyl-7α- (1-hydroxyethyl) -3-oxa-1-azabicyclo (4.2.0) octane 0.6 ml Powdered potassium hydroxide (19 mg, 0.332 mmol) was added to a solution of diethyl ether and after 15 minutes, 65 mg (0.302 mmol) of 4-nitrobenzyl chloroformate was added. The reaction mixture was then stirred at 25'C for 15 hours and then partitioned between 1M phosphate buffer (pH 7) and additional diethyl ether. The diethyl ether layer was washed with water, then brine, dried over magnesium sulfate and filtered. Concentration of the filtrate under reduced pressure gave 67 mg of the title compound as a colorless oil. The oil was purified by preparative thin layer chromatography on silica gel using benzene: ethyl acetate 9: 1 to afford the pure title compound as a diastereomeric mixture (40 mg).

IR spectrum (CH ₂ Cl ₂ ): µ 5.68 (beta-lactam ring and carbonate), 6.19 and 6.54 (nitro group)

Nuclear Magnetic Resonance Spectrum (CDCl ₃ ) δ 1.67 (d, 2H, Artf), 4.67 (s, ArCH ₂ ), 4.67-5.22 (m, CH ₃ CH), 5.98-6 , 25 (m, 2H, C-4 methylene), 6.25-6.62 (m, 1H, C-6 methine), 6.75-7.12 (m, 1H, C-7 methine), 7 , 75-8.83 (m, 2H, C-5 methylene), 8.22 (s, 3H, C-2 methyl) and 8.50-8.58 (m, 5H, C-2 methyl + Ctf ₃ CH).

The 7β-diastereoisomers or the mixture of 7α-7α and 7β-compounds can be prepared analogously.

Example 7 Preparation of trans-3- [1- (4-nitrobenzylcarbonyldioxy) ethyl] -4- (2-hydroxyethyl) -2-azetidinone (Scheme I) Dissolve in 1 ml of acetic acid and 2 ml of water. 0 g of 8-oxo-3-oxa-2,2-dimethyl-7α- [1- (4-nitrobenzylcarbonyldioxy) ethyl] -1-azabicyclo (4.2.0) octane and then The solution was kept at 65 ° C for 1.25 hours. The acetic acid and water were then removed under reduced pressure, and the residue was taken up in benzene and evaporated. This gives the title compound as a mixture of diastereoisomers.

IR (CH ₂ Cl ₂ ): µ 5.67 (beta-lactam ring), 5.72 (shoulder), 6.20 and 6.57 (nitro)

NMR (CDC1 _3): τ 1.73 (d, 2H, J = 8.5Hz, Ar-H), 2.43 (d, 2H, J = 8.5Hz, ARFIN), 3.63 (br s, 1H, NH), 4.37 to 5.13 (m, 1H, CH ₃ CH), 4.72 (s, 2H, _ArCH2), 6,07- 6.53 (m, 1H, methine CA); 6.23 (t, 2H, J = 5.5 Hz, CH ₂ OH),

6.73 to 6.93 (m, 1H, C-3 methine), 7.63 to 8.97 (m, 3H, CH ₂ CH ₂ OH), and 8.53 (d, J = 6.5 Hz, CH ₃ CH).

The cis-diastereoisomers or the cis-trans mixture can be prepared analogously.

8-11 below. in Examples 1 to 4 for the preparation of 3- [1 - (+ nitrobenzylcarbonyldioxy) ethyl] -4- (2-hydroxyethyl) azetidinone of Formula XXIII using one of the methods of Examples 4-7. Examples 4 to 6 are described in the Examples, or alternatively in the alternative.

Example 8 Preparation of 1- (2-Tetrahydropyranyl) -4- [2- (2-tetrahydropyranyloxy) ethyl] -2-azetidinone (Scheme J)

To a solution of 4- (2-hydroxyethyl) -2-azetidinone (62 mg, 0.539 mmol) in anhydrous p-dioxane (0.98 mL, 1.08 mmol) in a nitrogen atmosphere at 25 ° C was added a solution of 2,3-dihydropyran (0.98 mL, 1.08 mmol). and 19 mg (0.1 mmol) of p-toluenesulfonic acid monohydrate was added. The resulting reaction mixture was then stirred for 60 minutes and then partitioned between 10 mL of 0.5 M phosphate buffer pH 7 and 10 mL of ethyl acetate. The aqueous phase is then extracted with ethyl acetate and the combined organic phases are washed with brine, dried over magnesium sulfate and filtered. The filtrate was concentrated under reduced pressure to give 216 mg of crude product. The crude product was purified by preparative thin layer chromatography using ethyl acetate as the eluent. This gave 80 mg of the title compound as an oil.

NMR (CDC1 _3): τ 5.13 to 5.60 (m, OCH), 5,83- 6.85 (m, C-4H + _OCH2), 6.95 (dd, J = 5, 15 Hz, C-3 methylene), 7.35 (dd, J = 3 Hz and 15 Hz, C-3 methylene), and 7.62-8.95 (m, CHCH ₂ CH ₂ CH ₂ CH ₂ and CHCH ₂ CH ₂ O).

Example 9 Preparation of cis and trans -1- (2-Tetrahydropyranyl) -3- (1-hydroxyethyl) -4- [2- (2-tetrahydropyranyloxy) ethyl] -2-azetidinone (Scheme K)

When proceeding from 1- (2-tetrahydropyranyl) -4- [2- (2-tetrahydropyramyloxy) ethyl] -2-azetidinone, the same procedure as for 8-oxo-2,2-dimethyl-3-oxa-1- starting from azabicyclo (4.2.0) octane, 8-oxo-2,2-dimethyl-7α and p- (1-hydroxyethyl) -3-oxa-1-azabicyclo (4.2.0) octane the diastereomeric mixture gives the title cis and trans compounds.

-6181457

Example 10 Cis and trans 1- (2-Tetrahydropyranyl) -3- [1- (4-nitrobenzylcarbonyldioxy) ethyl] -4- [2- (2-tetrahydropyranyloxy) ethyl] -2-azetidinone Preparation (Scheme L)

If the procedure for trans-1- (2-tetrahydropyranyl) -3- (1-hydroxyethyl) -4- [2- (2-tetrahydropyranyl) oxyethyl] -2-azetidinone is the same as for 8-oxo-2 Starting from 2-dimethyl-7α- (1-hydroxyethyl) -3-oxa-1-azabicyclo (4.2.0) octane, 8-oxo-2,2-dimethyl-7α- [1- (4-nitro- benzylcarbonyldioxy) ethyl] -3-oxa-1-azabicyclo (4.2.0) octane gives the title trans compound. The diastereoisomers can be prepared analogously.

Example 11 Cis and trans -1- (3-tetrahydropyranyl) -3- [1- (4-nitrobenzylcarbonyldioxy) ethyl] -4- [2- (2-tetrahydropyranyloxy) ethyl] For the preparation of -2-azetidinone (Scheme M)

If the procedure for starting trans-1- (2-tetrahydropyranyl) -3- (1-hydroxyethyl) -4- [2- (2-tetrahydropyranyloxy) ethyl] -2-azetidinone is the same as for 8-oxo- Starting from 2,2-dimethyl-7a- (1-hydroxyethyl) -3-oxa-1-azabicyclo (4.2.0) octane, 8-oxo-2,2-dimethyl-7a- [1- (4) of nitrobenzylcarbonyldioxy) ethyl] -3-oxa-1-azabicyclo (4.2.0) octane gave the title trans compound. The cis-diastereoisomers can be prepared analogously.

Example 12 (5R, 6S, 8S) - and (5S, 6R, 8R) -2-Carboxy-3- (3-aminopropylthio) -6- (1-hydroxyethyl) -7-oxo-1- preparation of azabicyclo (3.2.0) hept-2-ene

Step A (Scheme N)

To a solution of 6.75 ml of anhydrous pyridine (molecular weight = 79, specific gravity 0.982 g / cm ³ , 83.9 mmol) in 350 ml of anhydrous acetonitrile was added 4.05 g (40.5 mmol, molecular weight = 100) of powdered chromium trioxide, and the resulting mixture was stirred for 30 minutes at 25 ° C. Then, 9.6 g of dried supercell filter material was added to the mixture, and the mixture was stirred for an additional 5 minutes. Thereafter, 3.21 g (9.5 mmol, 338 molecular weight) of trans-3- [1- (4-nitrobenzyl) carbonyldioxyethyl] -4- (2-hydroxyethyl) -2-azetidinone in 30 ml a solution of anhydrous acetonitrile was added all at once and the reaction mixture was stirred under anhydrous conditions at 25 ° C for 1 hour. 9.6 g of sodium hydrogen sulfate are then added to the reaction mixture, and after stirring for an additional 5 minutes, it is filtered on a bed of 40 g of silica gel and 40 g of anhydrous magnesium sulfate. The filter bed was then washed again with acetonitrile (600 mL total filtrate volume) and the filtrate was concentrated to 130 mL total volume under a nitrogen atmosphere.

The resulting crude aldehyde solution was then prepared at 0 ° C from 10.47 g (37) of 3- (4-nitrobenzyloxy) carbonylaminopropanethiol (270), as described in Example B). 7 mmol), followed by stirring the resulting reaction mixture with 8.0 ml of boron trifluoride-diethyl ether complex (MW = 142, specific gravity = 1.125 g / cm ³ ,

63.4 mmol). After stirring at 0 ° C for 1.5 hours, the reaction mixture was added with stirring to an ice-cold mixture of 69 g of potassium hydrogen phosphate, 500 ml of water and 700 ml of ethyl acetate. The phases were separated and the aqueous phase was extracted with sodium chloride and re-extracted with additional ethyl acetate. The combined organic layers were washed twice with brine, dried over anhydrous magnesium sulfate and filtered. The filtrate was concentrated under a nitrogen atmosphere and dried under high vacuum to give crude 1.

The crude product was purified by chromatography on a column of 450 g of silica gel, 48 cm high and 5.5 cm in diameter, eluting with chloroform, eluting with chloroform containing 0% to 4% methanol in increasing amounts. The fractions containing the desired product were combined, concentrated under a stream of nitrogen and finally dried under high vacuum. This gives compound 7.

Step B (Scheme 0)

While stirring in an ice bath, 600 g of diethyl ether and 75 ml of water were treated with 3.6 g (28.1 mmol) of 3-aminopropanetriol hydrochloride (molecular weight = 128), followed by 7.14 g (85 mmol) of sodium bicarbonate ( Molecular weight = 84) A solution of 75 ml of water was added. The ice bath was then removed and a solution of 6.75 g (31.3 mmol, molecular weight = 216) of chlorophenamic acid 4-nitrobenzyl ester in 270 ml of diethyl ether was added dropwise over 1 hour. After another 10 minutes, the phases are separated. the diethyl ether phase is extracted with 150 ml of 0.25 N hydrochloric acid and then with 200 ml of sodium chloride. Each aqueous extract was washed with diethyl ether (100 mL) and the organic layers were combined, dried over anhydrous magnesium sulfate, filtered, and concentrated in a nitrogen atmosphere. The crystalline residue was slurried with a small amount of diethyl ether, the resulting suspension was filtered and the pale yellow crystals were dried under high vacuum. This gives the desired compound.

Step C (Scheme P)

To 14.2 ml of pentane dried over a 4A Linde molecular sieve is added 0.5 ml of elemental bromine (molecular weight = 160.95.75 mmol). A solution of compound 1 (5.17 g, 6.02 mmol, molecular weight = 858) in a mixture of freshly distilled lithium aluminum hydride (58 mL) in tetrahydrofuran (65 mL), dried with diethyl ether (1.6mmol 3A Linde). At 0 [deg.] C., 10 ml of the aforementioned bromine solution (corresponding to 6.6 millimoles of bromine) are added dropwise under nitrogen atmosphere. After 10 minutes, 0.67 ml of cyclohexane (molar weight = 82, specific gravity = 0.81 g / cm ³ , 6.6 mmol) was also added at 0 ° C. After another 5 minutes, the reaction mixture was also cooled to 0 ° C

1.7 ml of triethylamine (molecular weight = 101, specific gravity = 0.729 g / cm ³ , 12.3 mmol) followed immediately by 40 ml of ice-cold dimethylformamide (distilled at 40 nunHg from anhydrous calcium sulfate and 4A Linde molecular sieve). above). The ice bath was then removed and stirring continued at room temperature for 2.25 hours. The reaction mixture was then poured into a stirred mixture of 12.6 mL of 1 M potassium hydrogen phosphate solution, 160 mL of water and 500 mL of ethyl acetate. After separation of the phases, the aqueous phase is saturated with sodium chloride, re-extracted with ethyl acetate, and the combined organic phases are extracted with sodium chloride, dried over anhydrous magnesium sulfate and concentrated to dryness under suction. dried. The crude compound 2 is thus obtained.

The crude compound 2 is then applied to a column of 250 g of silica gel, 45 cm long and 4.5 cm in diameter, in chloroform form, and in increasing amounts from 0% to 3%.

-7181457 was eluted with chloroform containing methanol. The fractions containing the pure product desired were combined, concentrated under a stream of nitrogen and dried under high vacuum to give pure 2.

The impure fractions from the elution can be rechromatographed on silica gel with chloroform containing ethyl acetate in increasing amounts of 0% to 25% to give additional 2.

Step D (Scheme S)

To a solution of 2.48 g (molecular weight = 388, 6.39 mmol) of bis (4-nitrobenzyl) keto-malonic acid ester prepared in Step E of the Example in 400 mL of hot anhydrous toluene is added 2 with stirring. A solution of 58 g of compound 2 (molecular weight = 588, 4.39 mmol) in a mixture of 20 ml of freshly distilled lithium aluminum hydride in 15 ml of anhydrous toluene. After removing some of the solvent by refluxing, additional anhydrous toluene was added to the reaction mixture and the azeotropic drying was repeated three more times. The resulting solution was then refluxed under nitrogen for 20 minutes and toluene was allowed to distil off so that no precipitate formed from the concentrated reaction mixture (total reflux time about 2.5 hours). The clear yellow solution thus obtained is then removed from the boiling oil bath and then placed under a stream of nitrogen gas where it is momentarily clouded. After evaporation to a yellow oil, the residue is dissolved in methylene chloride and the resulting solution is dried over anhydrous magnesium sulfate, filtered and concentrated under a nitrogen atmosphere to give crude 3 ash.

The crude product is then chromatographed on a column of 250 g of silica gel, 43 cm high and 4.5 cm in diameter, first with 500 ml of chloroform containing 0.5% methanol and then continuously with 1% methanol in chloroform. After elution of the reagents used predominantly, the fractions containing pure 3 were combined, evaporated under a stream of nitrogen and finally dried under high vacuum.

Subsequent chromatography of the remaining fractions containing the compound 3 and the fractions containing the corresponding cystiothenol ether on silica gel 40 gives an additional amount of compound 3.

Step E (Scheme T)

While stirring, 100 g (0.46 mol) of 4-nitrobenzyl bromide,

A mixture of 28.6 g (0.275 mol) of malonic acid and 750 ml of ethanol was heated in a 45 water bath until a homogeneous solution was obtained.

The solution is then carefully stirred with 33 g of potassium hydroxide (less than 85% purity: roughly 0.6 mol).

A solution of 200 ml of water was added followed by 200 ml of pure water and the resulting biphasic system was refluxed for 1.8 hours. The light-colored homogeneous solution was then cooled on ice for 1 hour, and the crude product was collected by filtration, washed twice with a minimum amount of cold ethanol, and blown dry with nitrogen to dry the filter cake. 33.7 g of a solid product are obtained. Conversely, if the reaction mixture is allowed to concentrate slowly to about half the volume while refluxing, a portion of the boiling solvent is distilled off, the yield of the crude product increases to 77 g.

The crude product is then recrystallized from methanol to give 60 ash pure bis (4-nitrobenzyl) ester of malonic acid.

A mixture of 23.4 g of compound Γ, 10 g of selenium dioxide and 30-40 ml of xylene is stirred in an oil-bathed flask and the temperature of the bath is raised to 130-135 ° C over 1 hour. The reaction mixture gradually becomes 65 and after stirring at 130-135 ° C for a total of 4 hours, most of the insoluble residue consists of black selenium. The reaction mixture was then cooled, magnesium sulfate was added to remove the water, and Celite was added to aid in filtration. The reaction mixture was filtered through a pad of Celite and the filter cake was washed with xylene and a small amount of ethyl acetate. The final volume of the filtrate is 60 ml. A 10g column of Baker silica gel with benzene was prepared and 10ml of the filtrate was applied to the column and eluted with increasing volumes of benzene containing ethyl acetate in 500 mL fractions. After separation of a benzene fraction containing 2% ethyl acetate and two benzene fractions containing 10% ethyl acetate, the third benzene fraction containing 10% and the first 20% ethyl acetate contained the bulk of the product (approximately 1.6 g). Separated from 10 ml of the filtrate) as shown by TLC on silica gel (20% ethyl acetate in chloroform as eluent). Recrystallize 1 g of the product from benzene by first dissolving it in 50 ml of benzene, concentrating the solution to about one third of its volume, and finally adding 1 ml of water-saturated benzene. 0.24 g of the bis (4-nitrobenzyl) ketomonic acid ester, m.p. 121-122 ° C and molecular weight 117, are obtained.

Step F (Scheme U)

A solution of compound 1.49 (molecular weight = 976, 1.53 mmol) in methylene chloride was dried over anhydrous magnesium sulfate, filtered, concentrated under a nitrogen atmosphere and dried under high vacuum just before the next reaction step. At -20 [deg.] C., to a solution of the compound 3 thus prepared in 24 ml of freshly distilled tetrahydrofuran from lithium aluminum hydride is added 0.206 ml of pyridine [molecular weight = 79, specific gravity = 0.982 g / cm ³ , 2.56 mmol]. A solution of 294 mg of freshly distilled thionyl chloride (119, 2.47 mmol) in 5 ml of tetrahydrofuran was added dropwise to the reaction mixture under nitrogen, and the reaction mixture was stirred at -20 ° C for 10 minutes at 0 ° C. Stir for 5 hours and at 25 ° C for 1 hour. The precipitated pyridine hydrochloride is then filtered off under a nitrogen atmosphere and washed with 20 ml of tetrahydrofuran. The filtrate was evaporated under a nitrogen atmosphere and the residue was suction dried under high vacuum. The resulting yellow foam is then stirred with 25 ml of anhydrous tetrahydrofuran and the small amount of orange insoluble material is filtered off under a nitrogen atmosphere. The filtrate was concentrated again to give a yellow foam again.

To the freshly prepared chloro compound was then first stirred, under stirring, 678 mg of a 9: 1 mixture of tributylphosphine (molecular weight = 202, 3.36 mmol) in water

36.5 ml of freshly prepared suspension are added followed by 294 mg of potassium hydrogen phosphate (molecular weight = 174.1.69 mmol). The reaction mixture was then stirred at 25 ° C for 35 minutes and then diluted with ethyl acetate (120 mL) and brine (60 mL). The phases are separated and the aqueous phase is extracted twice more with ethyl acetate. The combined organic layers were washed once with brine, dried over anhydrous magnesium sulfate, filtered, evaporated under nitrogen and suction dried under high vacuum. The crude compound 4 is thus obtained.

The crude material is then chromatographed on a column of 100 g of silica gel in chloroform, 28.5 cm high and 4 cm in diameter, the material to be purified is taken up in chloroform,

-8181457, then eluting with 0.5% methanol in chloroform. The fractions containing pure product were combined, evaporated under a nitrogen atmosphere and dried under high vacuum. The impure fractions can be rechromatographed on silica gel thin layers using a 1: 1 mixture of acetone-hexane as the eluent and the strips to be separated are extracted with chloroform / ethyl acetate. An additional amount of 4 may be obtained.

Step G (Scheme V)

To pentane (8.5 ml, dried over a 4A Linde molecular sieve) was added 0.2 ml elemental bromine (molecular weight = 160, 3.9 mmol). Subsequently, under nitrogen atmosphere at 0 ° C, 0.716 g of compound 4 (molecular weight = 960, 0.746 mmol) was dissolved in 18 ml of freshly distilled tetrahydrofuran and lithium aluminum hydride.

To a solution of 5.7 ml of diethyl ether dried over a 1.6 mm molecular sieve of Linde 3A is added dropwise 1.8 ml of the aforementioned 0.45 molar solution of pentane in bromine (corresponding to 0.81 mmol of bromine). . After stirring at 0 ° C for 15 minutes, the reaction mixture was treated with 0.42 ml of triethylamine (Mw = 101, specific gravity = 0.729 g / cm ³ , 3.03 mmol), followed immediately by 10.5 ml of ice-cold dimethylformamide. (distilled from calcium sulfate at 40 mmHg and stored above a 4A Linde molecular sieve). The ice bath was then removed and the reaction mixture was stirred at room temperature for 2 hours. The reaction mixture was then poured with stirring into an ice-cold mixture of 3.1 ml of 1 M potassium hydrogen phosphate solution, 70 ml of water and 100 ml of ethyl acetate. The layers were separated and the aqueous layer was saturated with sodium chloride and re-extracted with ethyl acetate. The combined organic layers were washed with brine, dried over anhydrous magnesium sulfate and filtered. The filtrate was concentrated under a nitrogen atmosphere and then dried under high vacuum. The crude compound 5 is thus obtained.

The crude material was chromatographed on a column of silica gel (60 g, 2.8 cm diameter) with chloroform followed by 100 ml of 2% ethyl acetate in chloroform, 100 ml of 4% ethyl acetate in chloroform and finally 5% ethyl acetate until the chromatography was completed. eluting with chloroform containing acetate. The pure product fractions were combined, evaporated under a nitrogen atmosphere and suction dried under high vacuum. This gives pure compound 5.

Step H (Scheme W) To a solution of mg of anhydrous silver fluoride (MW = 127, 0.23 mmol) in a solution of 148 mg (MW = 1038.0.14 mmol) in dry pyridine (3.5 mL) was added to the reaction mixture. The flask containing the flask was sealed and the contents stirred for one hour at room temperature in the dark and then poured into a mixture of 20 ml of cold water and 30 ml of ethyl acetate. After separation of the phases, the aqueous phase is extracted twice with ethyl acetate and once with chloroform. Each organic phase was extracted once with water and once with brine, and the combined organic phases were dried over anhydrous magnesium sulfate, filtered, concentrated in a nitrogen atmosphere and suction dried under high vacuum. The crude compound 6 is thus obtained.

This crude product was then subjected to preparative TLC purification using 40% acetone in hexane as the eluent and extracting the desired band with plenty of chloroform. This gives slightly contaminated compound 6. Further chromatography on silica gel (eluting with ethyl acetate-chloroform) afforded pure 6.

Step I (Reaction Scheme X)

To 13.4 mg (0.1 mmol) of anhydrous lithium iodide (molecular weight = 134; dried for several hours at 100 ° C under vacuum over phosphorus pentoxide) 78 mg of compound 6 (molecular weight = 958, 0.082 mmol) 0, 9 ml of powdered potassium hydroxide were distilled at 30 mmHg

A solution of S-collidine was added and the resulting reaction mixture was heated in a 120 ° C oil bath with stirring under nitrogen. After 30 minutes, the reaction mixture was cooled to 25 ° C, diluted with methylene chloride and poured into a round-bottomed flask where it was concentrated under a nitrogen atmosphere and then dried under high vacuum. The residue was then mixed with ethyl acetate, water and 1 mL of a 1M potassium hydrogen phosphate solution, the layers were separated and the aqueous layer was extracted twice with ethyl acetate and once with chloroform. Each organic phase is then washed with brine, combined, dried over anhydrous magnesium sulfate, filtered and concentrated under a nitrogen atmosphere. Finally, the resulting residue is vacuum dried under high vacuum to give crude 7.

On a silica gel plate, the crude compound was purified by preparative thin-layer chromatography, eluting with twice 40% acetone in hexane or extracting the bands to be separated with copious amounts of chloroform. 18 mg of starting material and 28 mg (44%) of 7 are obtained.

Step J (Scheme Y) to a solution of 7 mg (MW = 779, 0.064 mmol) in 0.7 mL of dimethyl sulfoxide (distilled from calcium hydride at 8 mmHg and stored over 4A Linde molecular sieve) for 100 μ 100 [MW = 101, specific gravity = 0.722 g / cm ³ , 0.71 mmol] diisopropylamine (distilled from sodium hydride under nitrogen atmosphere and stored above a 4A Linde molecular sieve) was added and the reaction flask was stirred for a few minutes and then left to stand for 2 hours. . Subsequently, the amine and most of the dimethylsulfoxide were removed under high vacuum without external heating, and the residue was quickly applied to a silica gel column (to which the residue was applied with ethyl acetate and eluted with the same solvent) to remove the remaining dimethylsulfoxide. After evaporation of all the ultraviolet absorbing fractions under a nitrogen atmosphere, the residue was purified by silica gel thin-layer chromatography, eluting with 50% ethyl acetate in chloroform as the eluent, to remove the ultraviolet light to isolate. This gave compound 8. The unreacted starting material can be reused or purified twice as described above to yield additional 8 compounds.

Step K (Scheme Z)

To 5.2 mg of compound 8 was added 0.6 ml of dioxane, 0.05 ml of ethanol, 0.35 ml of deionized water and 0.01 ml of 1.0 M potassium hydrogen phosphate solution, and 5 mg of the resulting clear solution were added. Palladium on carbon containing 10% of metal was added. The slurry was then purged with nitrogen gas and then alternately purged with hydrogen at 3.5 atm and vacuum for five or six times. Then the suspension

After shaking with hydrogen gas (3.5 atm) for 30-40 minutes, centrifuge and wash the catalyst two or three times with 0.5 ml to 0.5 ml deionized water and separate the washings by centrifugation. After centrifugation, the separated liquid phases are extracted five times with 1-2 ml of diethyl ether, and the

-9181457 was removed in heat and the aqueous solution was applied to a column of XAD-2 resin 20 times by 140 mm. Fractions of 100-100 drops (6-7 mL) were collected eluting with deionized water and the eluate was continuously exposed to ultraviolet light. The ultraviolet light absorbing material is shown in Figures 3-5. fractions begin to elute and usually occur at 25-30. fractions elute with fractions. Initially separated fractions are examined to exclude those that are excessively absorbed in the 270-280s region. The other fractions were combined and lyophilized. When this lyophilized material is dissolved in 10.0 mL of deionized water and is absorbed in ultraviolet light at 298 seconds, a yield of 10-30% of the desired product, compound 9, is observed.

Step L - Resolution of the racemic mixture

The desired (5R, 6S, 8S) - and (5S, 6R, 8R) -3- (2-aminopropylthio) -6- (1-hydroxyethyl) -7-oxo-1-aza-2- To a solution of the racemic mixture of carboxybicyclo (3.2.0) hept-2-ene in water containing 20% ethanol was added an equimolar amount of threo-S- (4-nitrophenyl) -2-aminopropane-1,3-diol and the resulting reaction mixture was heated to 50 ° C and then allowed to cool. When crystallization appears complete or complete, the mother liquor is carefully aspirated off and the remaining crystals are washed on the filter with a minimum amount of ice-cold 20% ethanol. The mixture of mother liquor and wash solution was then allowed to stand in the refrigerator for 18 hours when the other enantiomeric salt precipitated. This salt was collected by filtration and washed with a small amount of ice-cold water containing 20% ethanol. The salts are separately taken up with water and applied to columns made of XAD-2 resin, eluting the columns with water and observing the eluates under ultraviolet light. The aromatic amine is retained in the column while the desired isomers are present in the eluate and the desired (5R, 6S, 8S) or (5S, 6R, 8R) isomer is obtained by evaporation of the eluate.

The following example illustrates a preferred embodiment of the process of the invention when employed in a stereoselective manner. The starting material used is the 4-vinyl-2-azetidinone 23 in the form of the optically pure isomer which can be prepared as described below. This compound can thus be in the form of two isomers, i.e., 23 (5R) and 23 (5S) isomers. In the following example, each of the intermediates will be referred to by the above stereochemistry designation.

Example 13

The step

Preparation of 24 (5S) Compound, 1- (tert-Butyldimethylsilyl) -4-vinyl-2-azetidinone (Scheme AA)

To a solution of 1.153 g of compound 23 (5S) (11.89 mmol) and 1.82 ml of triethylamine (13.08 mmol) in anhydrous dimethylformamide at 0 ° C is added 1.885 g of tert-butyldimethylchlorosilane. (12.48 mmol) was added and a volumetric white precipitate immediately appeared. The precipitated mixture was then stirred for 1 hour while gradually warming to room temperature. The reaction mixture was then partitioned between methylene chloride (30 mL) and cold potassium dihydrogen phosphate (90 mL), and the aqueous layer was extracted with methylene chloride (20 mL). The combined organic phases are then washed four times with 30 ml each of water (30 ml) and finally with 50 ml of sodium chloride solution. The resulting methylene chloride solution was dried over anhydrous magnesium sulfate and filtered. The filtrate was concentrated under reduced pressure. 2.25 g of the title compound are obtained as a colorless liquid. NMR (CDC1 _3): τ 6.23 to 5.10 (m, ₂ Cfl == CFF), 4.07 (seven lines m, J = 8.6 and 3 Hz, C-477), 3, 35 (dd, J = 15 and 6 Hz, C-3 # to cis-C), 2.73 (dd, J = 15 and 3 Hz, C 3 H, trans to C 1/7), 0, 98 [s, (CH ₃ ) ₃ Si], 0.23 (s) and 0.18 [s, (CH ₃ ) ₂ Si],

The 24 (5R) isomer can be prepared as described above.

Step B)

The 25 (5R, 6S, 8R and S) and 25 (5R, 6R, 8R and S) compounds, i.e. 1- (tert-butyldimethylsilyl) -3- (1-hydroxyethyl) -4-vinyl Preparation of -2-azetidinone (Scheme B)

To a solution of freshly prepared lithium diisopropylamide (7.82 mmol) in anhydrous tetrahydrofuran (36 mL) under nitrogen atmosphere at -75 ° C was added a solution of 24 (5S) (1.5 g) in dry tetrahydrofuran (10 mL). After 16 minutes, the yellow solution of lithium enolate was treated with 1.59 mL of acetaldehyde (28.4 mmol). After a further 10 minutes, the reaction mixture was quenched with saturated aqueous ammonium chloride (30 mL) and extracted with ethyl acetate (50 mL, 25 mL). The combined organic layers were washed with brine (50 mL), dried over anhydrous magnesium sulfate, filtered, and concentrated in vacuo to give the crude product as a yellow oil. Chromatography on silica gel using 10% ethyl acetate in chloroform as the eluent gave the title isomeric mixture.

25 (5S, 6R, 8R and S) and 25 (5R, 6S, 8R and S) isomers can be prepared analogously.

Step C)

The 26 (5R, 6S, 8S) and 26 (5R, 6S, 8R) compounds, i.e., in particular the 1- (tert-butyldimethylsilyl) -3- [1- (4-nitrobenzyl) carbonyl] Preparation of ethyl] -4-vinyl-2-azetidinone (Scheme AC)

To a solution of 56 mg of 25 (5R, 6S, 8R) and 25 (5R, 6S, 8S) in 0.2 ml of anhydrous tetrahydrofuran in 2.4 ml of n-butyl lithium solution in hexane at -78 ° C-101 μΐ (0.242 mmol), and after 5 min, a solution of 52 mg of chloroformic acid 4-nitrobenzyl ester (0.242 mmol) in anhydrous tetrahydrofuran was added. After stirring at the same temperature for 55 minutes, saturated aqueous ammonium chloride (10 mL) was added to the reaction mixture, which was extracted with ethyl acetate. The extract was washed with brine, dried over anhydrous magnesium sulfate, filtered and concentrated in vacuo. This gave 91 mg of a yellow oil, which was purified by preparative thin layer chromatography on silica gel using 5% ethyl acetate in chloroform as eluent. This gave the title isomer mixture in a total yield of 54%.

Similarly, 26 (5R, 6R, 8R) and 26 (5R, 6R, 8R) and 26 (5R, 6R, 8R) and 26 (5R, 6R, 8R and S) diastereoisomers may be prepared in equivalent amounts to 25 (5R, 6R, 8R and S). compound.

The following diastereoisomers may also be prepared as described above:

(5S, 6R, 8R), (5S, 6R, 8S), (5S, 6S, 8R) and (5S, 6S, 8S).

Step D)

Preparation of 27 (5R, 6S, 8R) Compound 3- [1- (4-Nitrobenzyl) -carbamoyl-10181457-Bonyloxyethyl] -4-vinyl-2-azetidinone (Scheme AD).

At room temperature (25 ° C), a solution of 26 mg (5R, 6S, 8R) (61 mg) (0.141 mmol) in 2 mL of 0.5 N HCl was stirred for 3 hours, then cooled to 0 ° C and 5 mL of 5% neutralized by the addition of aqueous sodium bicarbonate solution. The solution is then extracted twice with 5 ml of ethyl acetate and the combined extracts are washed twice with 5 ml of water and once with 10 ml of brine, dried over anhydrous magnesium sulfate, filtered and the filtrate is evaporated in vacuo to an oil. The latter was subjected to preparative thin layer chromatography on silica gel, eluting with 10% ethyl acetate in chloroform. 44 mg of the title compound are obtained. 15

The following isomers can be prepared from the appropriate starting materials as described in Step D:

(5R, 6R, 8R) -> · 27 (5R, 6R, 8R) (5R, 6R, 8S) - 27 (5R, 6R, 8S) 20 (5R, 6S, 8S) - 27 (5R, 6S, 8S) ) (5S, 6R, 8R) - * 27 (5S, 6R, 8R) (5S, 6R, 8S) - 27 (5S, 6R, 8S) (5S, 6S, 8R) -> 27 (5S, 6S, 8R) ) (5S, 6S, 8S) - 27 (5S, 6S, 8S) 25

Step E)

Preparation of 28 (5S, 6S, 8R) (Scheme AE)

A solution of 101 mg of bis (4-nitrobenzyloxycarbonylaminopropyl) disulfide (0.19 mmol) in 1.5 ml of lithium aluminum hydride in freshly distilled tetrahydrofuran was cooled to 25 ° C and then while stirring, 0.5 ml (corresponding to 0.19 mmol of bromine) is added dropwise to a total volume of 2.2 ml of a solution of carbon tetrachloride (135 mg) dried over an elemental bromine molecular sieve.

The resulting orange solution was stirred at -20 ° C for 5 min and then a solution of 54 mg of (5R, 6S, 8R) in methylene chloride (0.5 mL) dried over a molecular sieve. The color of the reaction mixture turns yellow. The reaction mixture was then allowed to warm to 0 ° C over 5-10 minutes.

Thin layer chromatography (silica gel, eluting with 5% methanol in methylene chloride or 20% ethyl acetate in methylene chloride as eluent) showed a major spot which was R _f and Ce ⁴⁺ (H ⁺ ) - is different from either the disulfide or the starting 4-vinyl-2-azetidine 45 non-ion. The reaction mixture was concentrated to 0.5 mL under a nitrogen atmosphere, applied directly to two strips of 203 x 203 mm silica gel (1000 microns thick) and run with 20% ethyl acetate in methylene chloride. The 50 major bands which become visible under ultraviolet light are scraped off and extracted with ethyl acetate. The title compound was obtained.

Step F)

Preparation of Compound 14 (5S, 6S, 8R) (Scheme AF): Mixing 28 mg (5S, 6S, 8R) (0.162 mmol) in 1.0 mL of 55 mL of calcium hydride distilled over molecular sieve with stirring under nitrogen 25 DBU (0.19 mmol) were added. After 3 hours, the reaction mixture was poured into aqueous potassium dihydrogen phosphate solution and extracted several times with ethyl acetate. The combined extracts were washed twice with water, dried over anhydrous magnesium sulfate and evaporated under a nitrogen atmosphere. The crude product (42 mg) was applied 203 times to 203 mm and 1000 μg silica gel plates and run on 20% ethyl acetate in methylene chloride. The title compound was obtained.

The following isomers may be prepared analogously:

(5R, 6R, 8R), (5R, 6R, 8S), (5R, 6S, 8S), (5S, 6R, 8R), (5S, 6R, 8S), (5R, 6S, 8R) and (5S) , 6S, 8S).

Step G)

Preparation of the various isomers of the compound of formula (I)

By following the procedure described in Steps D-K of Example 12, but of course starting from the corresponding isomers of compound 14, the following compounds can be prepared:

I (5R, 6R, 8R), I (5R, 6R, 8S), I (5R, 6S, 8R), I (5R, 6S, 8S),

I (5S, 6S, 8S),

I (5S, 6S, 8R),

I (5S, 6R, 8S) and I (5S, 6R, 8R).

Example 14 Preparation of bis (4-nitrobenzyloxycarbonylaminopropyl) disulfide (Scheme AG)

To a solution of 3- (4-nitrobenzyloxycarbonylamino) -propanethiol (11.9 g, 0.044 mmol) in 100 mL of anhydrous tetrahydrofuran at -20 ° C under nitrogen is added 1.21 mL (0.022 mmol) of basic bromine and the cooling bath was removed and the cold solution was stirred for 15 minutes. The solution was then diluted with 400 mL of ethyl acetate, successively with 200 mL of 1 M phosphate buffer, pH 7.0, 200 mL of 1 M potassium dihydrogen phosphate, 200 mL of water, 100 mL of water and finally 200 mL of sodium chloride. chloride solution. The solution was dried over anhydrous magnesium sulfate, filtered, and the filtrate concentrated in vacuo. The resulting yellow precipitate was chromatographed on silica gel, eluting with 5% ethyl acetate in chloroform. The title compound is crystallized.

IR (CH ₂ Cl ₂ ): µ 3.04 (NH), 5.96 (carbonyl), 6.22 and 6.61 (nitro)

Example 15

In the same manner as in the preceding and following preparative examples, the following compounds of formula I can be prepared (the first compound of the table is the compound of Examples 12 or 13):

Serial number of compound R ' R ¹ —SR ⁸ con- figu- rationality tion λ max NMR Spectrum (shifts in δ) 8 bety- constitution CH 5 helyzetfi CH Other 1 CH ₃ CH (OH) - H -S (CH ₂ ) ₃ NH ₂ 305 1.23 4.07 CH ₂ Ctf ₂ NH ₂ : 3.05 0 CH ₃ CH (OH) - H 3-pyridylthio 300

-11181457

The sot number of the compound R ' R ⁷ SR ' con- figu- rationality tion λ max NMR Spectrum (δ shifts) 8-position ch ₃ 5 at positions CH Other 3 4 5 6 7 8 CH ₃ CH (OH> - CH ₃ CH (OH) - CH ₃ CH (OH) - CH ₃ CH (OH) - CH ₃ CH (OH) - CH ₃ CH (OH) - H H H H H H —Sch ₂ ch ₃ —SCH (CH ₃ ) ₂ (sodium salt) —SCH ₂ CH ₂ OH —sch ₂ cooch ₃ —SCH ₂ COH o 1 —sch ₂ ch ₂ nh ₂ 292.2? 8 297 295 297 303 287.289 9 CH ₃ CH (OH) - H —SCH ₂ C (CH ₃ ) ₂ NH ₂ NH II nhcnh ₂ I 298 1.29 4.22 (CH ₃ ) ₂ -C: 1.44 10 CH ₃ CH (OH) - H 1 —S — CH ₂ —CH — CH ₃ ch ₂ —nh ₂ I R 300 1.34 3.43 CH ₃ -CH-: 1.28, 1.35 11 12 CH ₃ CH (OH) - CH ₃ CH (OH) - H H 1 -S CH-CH ₂ CH ₃ -S-CH ₂ C (CH ₃ ) ₂ CH ₂ NHCH = NH NH II R, S 297 300 1.28 4.2-4.3 CH ₂ CH ₃ : 1.06 13 CH ₃ CH (OH) - H II —SCH ₂ C (CH ₃ ) ₂ NHCNH ₂ - 1.23 - (CH ₃ ) ₂ -C-: 1.43 14 15 CH ₃ CH (OH) - CH ₃ CH (OH) - H H —S — CH (CH ₃ ) —ch ₂ —nh ₃ ⁺ NH 2 CH ₂ —NHCH 1 —S — CH — CH ₂ —CH ₃ NH II R, S R, S 296 300 1.28 4.23 CH-CH3: 1.45 16 CH ₃ CH (OH> - H II —SCH ₂ CH (CH ₃ ) NHCH NH II R 298 1.30 4.23 CH-CH ₃ : 1.5 17 CH ₃ CH (OH) - H II —SCH ₂ CH (CH ₃ ) NHCNH ₂ S 300 1.22 4.11 CH-Cff ₃ : 1.13 18 19 20 21 22 CH ₃ CH (OH) - CH ₃ CH (OH> - CH ₃ CH (OH) - CH ₃ CH (OH) - CH ₃ CH (OH) - H H H H H -SCH ₂ C (CH _{3) 2} CH ₂ NH _2, -SCH ₂ CH ₂ COONa -SC (NH ₂₎ COOH, -SCH ₂ COOCH ₂ NH fennil II -SCH ₂ CH ₂ CH ₂ NH ₂ NHCNH || R 301 301 300 300 303 1.33 4.1 (CH ₃ ) ₂ -C-: 1.03 23 CH ₃ CH (OH) - H II —SCH (CH ₃ ) CH ₂ NHCNH ₂ R, S 298 1.30 4.24 CH-CH3: 1.36, 1.42 24 CH ₃ CH (OH) - H —SCH (CH ₃ ) CH ₂ NH ₃ ⁺ NH II RorS 296 1.31 4.25 CH-Cff ₃ : 1.42 25 CH ₃ CH (OH) - H II —SCH (CH) ₃ CH ₂ NHCH RorS 298 1.28 4.23 CH-CH ₃ : 1.36 26 CH ₃ CH (0H) - H —SCH ₂ CH (CH ₃ ) NH ₂ NH II CH ₂ NHCH 1 S 297 1.33 4.27 CH ₃ -CH: 1.43 27 CH ₃ CH (OH) - H 1 —S — CH — CH ₂ CH ₂ CH ₃ NH II R 298 1.32 4.1-4.3 CH ₂ CH ₃ : 0.96 78 CH ₃ CH (OH) - H II —SCH (CH ₃ ) CH (CH ₃ ) NHCH R, S ' 298 1.26 4.20 CH-CH ₃ : 1.32, 134

-12181457

Serial number of compound R ^ö R ' —SR ⁸ con- figu- rationality tion λ max NMR Spectrum (δ shifts) 8-position ch ₃ 5-position CH Other NH II —SCH (CH ₃ ) CH (CH ₃ ) NHCH NH II —SCH (CH ₃ ) CH ₂ N (CH ₃ ) CH —S (CH ₂ ) ₂ NH ₂ 29 CH ₃ CH (CH) - H R, S ²⁾ 299 30 31 CH ₃ CH (CH) - ohch ₂ H CH ₃ 298 298 1.28 4.26 CH-CH ₃ : 1.33, 1.34

¹ the two methyl groups are in the same spacer ² the two methyl groups are in the same spacer Example 16

Preparation of Pharmaceutical Compositions Such a dosage unit is obtained when 3-benzylthio-2-carboxy-6- (1'-hydroxyethyl) -7-oxo-1-azabicyclo (3.2.0) -hept-2-ene is obtained with mg of lactose. and an equimolar mixture of 5 mg of magnesium stearate (total weight 145 mg) in gelatin capsules # 3. Similarly, but using, for example, more active ingredients and less lactose, gelatin capsules with a total content of 145 mg can also be prepared. If we want to encapsulate more than 145 mg in total, larger capsules are utilized.

For other pharmaceutical formulations, the following prescription-like formulations are provided.

Tablet

component

Quantity per tablet

3-Benzylthio-2-carboxy-6- (1'-hydroxyethyl) -7-oxo-1-azabicyclo (3.2.0) hept-2-ene corn starch ¹ dicalcium phosphate lactose ¹ magnesium stearate

125 mg 6 mg

192 mg 192 mg 800 mg US Pharmacopoeia The active ingredient is mixed with dicalcium phosphate, lactose, and cornstarch. The mixture was then granulated with 15% corn starch paste (6 mg) and subjected to coarse sieving. The resulting pellets were dried at 45 ° C and sieved through a 16 mesh screen. The corn starch residue and the magnesium stearate are then added to the granules and the resulting blends are compressed into 800 mg tablets with a diameter of approximately 10 mm. Parenteral Solution (wrapped ampoules) of 3-benzylthio-2-carboxy-6- (1-hydroxyethyl) for injection ¹⁵⁰⁰ mg -7-oxo-1-azabicyclo (3.2.0) hept-2-ene sterile water 2 ml

Eye drops

3-Benzylthio-2-carboxy-6- (1'-hydroxyethyl) -7-oxo-1-azabicyclo (3.2.0) hept-2-ene 100 mg hydroxypropylmethylcellulose 5 mg sterile water 1 ml required

ear drops

3-Benzylthio-2-carboxy-6- (1'-hydroxyethyl) -7-oxo-1-azabicyclo (3.2.0) hept-2-ene 100 mg benzalkonium chloride 0.1 mg sterile water 1 ml required

Topical ointment

3-benzylthio-2-carboxy-6- (l'-hidroxi25 ethyl) -7-oxo-1-azabicyclo (3.2.0) hept-2-ene 100 mg Polyethylene glycol 4000 ¹⁴⁰⁰ mg Polyethylene Glycol 400 '1, 0 g US Pharmacopoeia quality

In the above formulations, the active ingredient may optionally be used in combination with other known biologically active components such as Lincomycin, a penicillin, Streptomycin, Novobio35 cin, Gentamicin, Neomycin, Colistin or KanamyCin, or other therapeutic compounds such as Probenicid. added.

Example 17

Preparation of 8-oxo-2,2-dimethyl-7a-isopropyl-3-oxa-1-aza-bicyclo (4,2,0) octane (Compound XIX) in THF under N ₂ 1.54 ml diisopropyl 45 and cooled to -78 ° C. 5.97 ml of 1.97 moles of n-butyllithium in hexane are added dropwise over 5 minutes. The reaction mixture was stirred at -78 ° C for 10 minutes and then 1.55 g of 8-oxo-2,2-dimethyl-3-oxa-1-azabicyclo (4,2,0) octane in 15 ml of THF was added. every 5 minutes. After another 10 minutes, 1.97 mL of hexamethylphosphoramide was added. The mixture was stirred for an additional 10 minutes and then treated with isopropyl iodide (ml). The reaction mixture was stirred at -78 ° C for 15 minutes, allowed to warm to 25 ° C and stirred for 15 minutes. The reaction mixture was diluted with EtOAc, washed once with pH 7 phosphate buffer, dried and evaporated. The residue was chromatographed on silica gel eluting with 25% EtOAc-C ₆ H ₆ to give 8-oxo-2,2-dimethyl-7a-isopropyl-3-oxa-1-azabicyclo (4,2,0) -octane. μ ir: 5.7 (β-lactam).

NMR δ: 0.96d, 1.06d (CH ₃ -C-Η): 1.4S, 1.76S (gem dimer ch ₃ til);

1.9m (C-5H); 2.59-d (C-7);

3.33m (C-6H); 3.83-d (C- ^ H).

-1 327

Example 18

Preparation of 8-oxo-2,2,7-trimethyl-3-oxa-1-azabicyclo (4,2,0) octane (XX)

A17. The procedure of Example 1b is followed but the isopropyl iodide is replaced by an equivalent amount of methyl iodide to give the desired compound.

Example 19

Preparation of 8-oxo-2,2,7-trimethyl-7- (hydroxymethyl) -3-oxa-1-azabicyclo (4,2,0) octane (Scheme AH)

1.1 eq of freshly prepared lithium diisopropylamide was dissolved in anhydrous tetrahydrofuran at -78 ° C under nitrogen, then 8-oxo-2,2,7-trimethyl-3-oxa- dissolved in anhydrous tetrahydrofuran and cooled to -78 ° C. 1-Azabicyclo (4,2,0) octane is added. After two minutes, the resulting lithium enolate is treated with excess formaldehyde, which is supplied in gaseous form over the surface of the solution with stirring. The mixture was stirred for an additional 30 minutes at -78 ° C and poured into water. The aqueous phase was saturated with sodium chloride and extracted with ethyl acetate. The combined ethyl acetate solutions were dried over magnesium sulfate and filtered. The filtrates were evaporated under reduced pressure to give crude product. This was purified by silica gel chromatography using ethyl acetate-benzene. 8-Oxo-2,2,7-trimethyl- (hydroxymethyl) -3-oxa-1-azabicyclo (4,2,0) octane is obtained.

Example 20

Preparation of 8-oxo-2,2,7-trimethyl-7- (p-nitrobenzylcarbonyldioxymethyl) -3-oxa-1-azabicyclo (4,2,0) octane (Scheme A) mg (0.302 mmol) 8-oxo A solution of -2,2,7-trimethyl-7- (hydroxymethyl) -3-oxa-1-azabicyclo (4,2,0) octane in 0.6 ml of ether under anhydrous conditions at 0 ° C was 19 mg (0.332 mmol). with powdered potassium hydroxide. After 15 minutes, 65 mg (0.302 mmol) of nitrobenzyl chloroformate was added to the reaction mixture. Stirring was continued at 25 ° C for 15 hours. The mixture was partitioned between 1M phosphate buffer pH 7 and several ethers. The ether layer was washed with water and brine, dried over magnesium sulfate and filtered. Concentration of the filtrate under reduced pressure gave 67 mg of a colorless oil. Purification was by thin layer chromatography on silica gel eluting with 1: 9 acetate-benzene. 40 mg of 8-oxo-2,2,7-trimethyl-7- (p-nitrobenzylcarbonyldioxymethyl) -3-oxa-1-azabicyclo (4,2,0) octane are obtained in the form of a diastereomeric mixture.

Example 21

Preparation of 3-methyl-3- (p-nitrobenzylcarbonyldioxymethyl) -4- (2-hydroxyethyl) -2-azetidinone (Scheme AJ) Dissolve 1.0 g of 8-oxo-3- in acetic acid and 2 ml of water. oxa-2,2,7-trimethyl-7- (lp-nitrobenzyl dioxymethylene-carbonyl) -l-azabicyclo (4,2,0) octane. The acetic acid and water were removed under reduced pressure and the residue taken up in benzene and evaporated. 3-Methyl-3- (p-nitrobenzylcarbonyl-dioxymethyl) -4- (2-hydroxyethyl) -2-azetidinone was obtained as a diastereomeric mixture.

Example 22

Preparation of 3-methyl-1- (2-tetrahydropyranyl) -4- [2- (2-tetrahydropyranyl) oxyethyl] -2-azetidinone

The title compound can be prepared from the compound prepared in Example 8 by the procedure described in Example 21.

Example 23

Preparation of (2-tetrahydropyranyl) -3-methyl-3- (1-hydroxymethyl) -4- [2- (2-tetrahydropyranyl) oxyethyl] -2-azetidinone (Scheme AK)

Following the procedure described for the preparation of 8-oxo-2,2,7-trimethyl-7- (hydroxymethyl) -3-oxa-1-azabicyclo (4,2,0) octane, 8-oxo-2,2,7-trimethyl-3 oxa-1-azabicyclo (4,2,0) octane (Example 19 above) and 3-methyl-1- (2-tetrahydropyranyl) -4- [2- (2-tetrahydropyranyl) oxyethyl] -2-azetidinone yielding a diastereomeric mixture of 1- (2-tetrahydropyranyl) -3-methyl-3- (hydroxymethyl) -4- [2- (2-tetrahydropyranyl) oxyethyl] -2-azetidinone.

Example 24

Preparation of 3-methyl-1- (2-tetrahydropyranyl) -3- (1-p-nitrobenzylcarbonyldioxymethyl) -4- [2- (2-tetrahydropyranyl) oxyethyl] -2-azetidinone (Scheme L)

8-Oxo-2,2,7-trimethyl-7- (1-p-nitrobenzylcarbonyl-dioxymethyl) -3-oxa-1-azabicyclo (4,2,0) octane 8-oxo-2,2,7-trimethyl Following the procedure described for the preparation of -7- (1-hydroxymethyl) -3-oxa-1-azabicyclo (4,2,0) octane, and 3-methyl-1- (2-tetrahydropyranyl) -3- (hydroxymethyl) -4 Using [2- (2-tetrahydropyranyl) oxyethyl] -2-azetidinone, 3-methyl-1- (2-tetrahydropyranyl) -3- (p-nitrobenzylcarbonyldioxymethyl) -4- [2- (2-tetrahydropyranyl) is obtained. -oxietil] -2-azetidinone.

Example 25

Preparation of 3-methyl-3- (p-nitrobenzylcarbonyldioxymethyl) -4- (2-hydroxyethyl) -2-azetidinone (Scheme AM)

A solution of 3-methyl-1- (2-tetrahydropyranyl) -3- (p-nitrobenzylcarbonyldioxymethyl) -4- [2- (2-tetrahydropyranyl) oxyethyl] -2-azetidinone in methanol at 25 ° C was 0.1 mol. equivalent p-toluenesulfonic acid monohydrate. The mixture was stirred for 2 hours and then neutralized with 1M phosphate buffer pH 7. The product was extracted with ethyl acetate. The ethyl acetate solution was washed with brine, dried over magnesium sulfate and filtered. The filtrate was concentrated under reduced pressure to give 3-methyl-3- (p-nitrobenzylcarbonyldioxymethyl) -4- (2-hydroxyethyl) -2-azetidinone.

Example 26

Preparation of 3- (2-aminoethylthio) -6-methyl-6- (hydroxymethyl) -7-oxo-1-azabidochloro (3,2,0) hept-2-ene-2-carboxylic acid (Scheme AN)

The step

To a solution of 6.75 ml (83.9 mmol) of anhydrous pyridine (molar weight 79, weight 0.982) in 350 ml of anhydrous acetonitrile is added 4.05 g (40.5 mmol) of anhydrous chromium trioxide (100 molar weight). After stirring for 30 minutes at room temperature (25 'C), 9.6 g of dry Supercell are added and stirring is continued for another 5 minutes. Then 3.21 g (9.5 mmol) of 3-methyl-3- (p-nitrobenzyloxycarbonyldioxymethyl) -5- (2-hydroxyethyl) -2-azetidinone dissolved in 30 ml are added simultaneously. The reaction mixture is anhydrous

After stirring at room temperature (25 ° C) for one hour, sodium bisulfite (9.6 g) was added and stirring continued for another 5 minutes. It was then filtered through a column containing 40 g of silica gel and 40 g of dry magnesium sulfate, and the column was washed several times with approximately 600 ml of acetonitrile. The filtrate was concentrated to 130 mL under N ₂ . To this crude aldehyde solution is added p-nitrobenzyloxycarbonylaminoethane thiol (9.64 g, 37.7 mmol) at 0 ° C under N ( ₂ ). Boron trifluoride etherate (8.0 mL, 63.4 mmol) (MW 142, density 1.125) was added with stirring. After 1.5 hours at 0 ° C, the mixture was poured into a ice-cold mixture of 69 g of K ₂ HPO in _{4 to} 500 ml of water and 700 ml of ethyl acetate (EA) with stirring. The resulting layers were separated, the aqueous layer was saturated with NaCl and re-extracted with additional ethyl acetate. The combined organic layers were washed twice with brine, dried over anhydrous magnesium sulfate, and filtered. The filtrate was concentrated under N ₂ and then concentrated under high vacuum to give crude 12/1.

The product was chromatographed on 450 g of silica gel (column height 48 cm, diameter 5.5 cm) using CHCl _{3 and} eluted with CHCl ₃ containing increasing amounts of metal hydroxide (MeOH) (0-4% MeOH / CHCl ₃ ). The fractions containing the desired product were combined, concentrated in a stream of N ₂ and evaporated under high vacuum to give 12/1.

Step B: Preparation of p-nitrobenzyloxycarbonylaminoethane thiol (Scheme AO)

To a mixture of diethyl ether (600 ml) and water (75 ml) was added 3.2 g (28.1 mmol) of cysteamine hydrochloride (molar weight 114) in an ice bath. To the solution was added sodium hydrogencarbonate (7.14 g, 85 mmol) in water (75 mL). The ice bath was removed and 6.75 g (216 mmol) of p-nitrobenzyl ester of chloroformic acid (molar weight 216) in diethyl ether (270 mL) was added dropwise over about 1 hour. 10 minutes after the addition was complete, the layers were separated. The ether layer was extracted with 150 mL 0.25 N HCl followed by 200 mL brine. The aqueous layers were washed thoroughly with 100 mL of diethyl ether. The combined ethereal phases are dried over anhydrous magnesium sulfate, filtered and concentrated in a stream of N ₂ . The crystalline residue was liquefied with a small amount of ether, filtered and the resulting slightly yellow crystals dried under high vacuum. 4.7 g of p-nitrobenzyloxycarbonylaminoethanethiol are obtained in a yield of 65%.

NMR (CDCl3) 8.18 (d, J = 8Hz, aromatic protons ortho to the nitro group), 7.47 (¢ /, J = 8Hz, aromatic protons in the nitro group), 5.27 (- Nfí-), 5.20 (.V, -CH ₂ - p-nitrophenyl), 3.40 (m, - CH ₂ -NH -), 2.67 (m, _-CH2 -SH), 1.35 (?, J = 8.5Hz, -SA) from TMS IR (CHCl ₃ solution) carbonyl group ~ 1725 cm ⁴ MS - Molecular ion-256, (M-47) 209 (M-136) 120- p-nitrobenzyl at 136

Step C (Scheme AP)

To 14.2 g of pentane dried by Linde molecular sieve is added 0.5 ml (9.75 mmol) of bromine. 5 g (6.02 mmol) of 12/1 (molar weight = 830) was dissolved in lithium aluminum hydride (LAH) in freshly distilled 58 ml of tetrahydrofuran (THF) and 65 ml of diethyl ether anhydrous on a Linde molecular sieve 3A1 / 16. To this solution was added dropwise 10 ml (6.6 mmol) of the above 0.67 molar bromine solution at 0 ° C with stirring under nitrogen. The resulting mixture was allowed to stand at 0 ° C for 10 min, and after 5 min 0.67 mL (6.6 mmol) cyclohexane (molar weight = 82, density = 0.81) was added at 0 ° C. After a further 5 minutes at 0 ° C, 1.7 ml (12.3 mmol) of triethylamine (molar weight 101, density = 0.729) were added, followed immediately by 40 ml of ice-cold dimethylformamide (DMF) which was distilled from anhydrous CaSO ₄ . It was stored under Hgmm pressure and over a 4A Linde molecular sieve. After the addition was complete, the ice bath was removed and stirring was continued at room temperature for a further 2 and a / 4 hours. The reaction mixture was then stirred with 12.6 mL of a 1 M solution of KH ₂ PO ₄ , 160 mL of water and 500 mL of ice-cold (EA). After separation of the layers formed, the aqueous phase is saturated with sodium chloride and extracted again with EA. The combined organic planks lot ^of extracted once with brine, dried over anhydrous magnesium sulfate, filtered and concentrated under _N2 stream. The concentrate was concentrated under high vacuum to give compound 12/2.

The product was chromatographed on a 45 cm high, 4.5 cm diameter column containing 250 g of silica gel in CHCl ₃ and eluted with CHCl ₃ containing increasing proportions of MeOH. (0-3% MeOH / CHCl ₃ ). The fractions containing the pure product were collected, concentrated in a stream of N ₂ and dried under high vacuum. The impure fractions were re-chromatographed on silica gel to give compound 12/2, containing increasing amounts of EA in CHCl ₃ (0-25% EA / CHCl ₃ ).

Step D (Scheme AQ)

2.48 g (6.39 mmol) of di- (p-nitrobenzyl) ketomalonate (MW 388) prepared in Step E of Example 12 are dissolved in 400 mL of hot anhydrous toluene and with stirring in 20 mL of THF (from LAH). distilled) and 2.52 g (4.39 mmol) of compound 2 dissolved in 40 ml of anhydrous toluene are added. After boiling a portion of the solvent, additional anhydrous toluene was added and the azeo-drying procedure was repeated three times. The solution was then heated to reflux for three hours. Further toluene is boiled until the volume of the solution decreases until a precipitate begins to form. The total heating time is about 2 to% hours. After removing the clear yellow reaction mixture from the oil bath, it is placed under an atmosphere of N ₂ , which immediately causes turbidity. After concentrating to a yellow oil consistency, the residue was dissolved in CH ₂ Cl ₂ and dried over anhydrous magnesium sulfate, filtered and concentrated under N _{2 to} give (12/3).

The material is 43 cm high and filled with 250 g of silica gel

Chromatograph on a 4.5 cm column using CHCl ₃ . Elution with 500 mL of 0.5% MeOH / CHCl ₃ followed by elution with 1% MeOH / CHCl ₃ . After the appearance of excess reagent, the fractions containing the compound (12/3) were collected and concentrated in a stream of N ₂ and then dried under high vacuum.

Subsequent fractions containing the compound (12/3) and the corresponding cysteiene ether were rechromatographed on silica gel to give additional compound (12/3).

Step E)

Preparation of di-p-nitrobenzyl ketomalonate (Scheme AR)

A mixture of p-nitrobenzyl bromide (100 g, 0.46 mol), malonic acid (28.6 g, 0.275 mmol) and ethanol (750 ml) was heated with stirring in a steam bath until a clear solution was obtained. 33 g (about 0.6 mol) of KOH (85 g) in water (200 mL) are added gently while stirring. An additional 200 ml of water was added and the biphasic system was refluxed for 1.8 hours.

-15181457. The slightly colored homogeneous solution was cooled with ice for 1 hour and the crude product was isolated by filtration. It is washed twice with a minimum amount of cold ethanol and dried on a filter cake by flowing dry nitrogen. 33.7 g of a solid are obtained. If heated at reflux, the reaction mixture is allowed to concentrate slowly to about a half volume by evaporation of the solvent, yielding crude product to 77 g.

The product is recrystallized from methanol to give the pure di-p-nitrobenzylmalonate (12/1 '). 10

A mixture of 23.4 g of 12/1 ', 10 g of SeO ₂ and 30-40 ml of xylene is stirred in an oil bath flask.

The bath temperature was raised to 130-135 ° C over 1 hour. Meanwhile, a gradual darkening of the reaction mixture is observed. After heating for 4 hours at 130-135 ° C, most of the insoluble residue is black selenium.

The mixture was cooled, the water extracted with MgSO ₄ and "Celite" added to facilitate filtration. The mixture was filtered through Celite, and the filter cake was washed with xylene and a small amount of EtOAc. Final volume 60 ml. A 10g Baker 20 silica gel column was prepared with benzene and passed through 10 ml of filtrate. Elute with increasing amounts of benzene containing EtoAc and collect 500 ml fractions. After removal of a 2% ethyl acetate / benzene fraction and two 10% EtOAc benzene fractions, the third 10% and 25 first 20% EtOAc / benzene fractions contained the bulk of the product (1.6 g). , from the 10 ml filtrate) as estimated by TLC (20% EtOAc / CHCl ₃ ; silica gel GF). Recrystallization from benzene (1 g in 50 mL and concentrated to 1/3 volume and initialized with 30 mL of saturated benzene in water) gives 0.24 g of 12/2 '. Op: (117) 121-122 'CF Step (AS Scheme)

1.468 g (1.53 mmol) of 12/3 (molar weight 962) are dissolved in CH ₂ Cl ₂ - 35 dried over anhydrous magnesium sulfate, filtered, concentrated in a stream of N ₂ and further dried under high vacuum until use. Dissolve 12/3 in 24 mL of THF (freshly distilled from LAH) and add 0.206 mL (2.56 mmol) of anhydrous pyridine (Mw = 79, density = 0.982) at -20 ° C. While stirring under 40 N ₂ atmosphere, 249 mg (2.47 mmol) of freshly distilled thionyl chloride (molar weight = 119) are added dropwise. The reaction mixture was then stirred at -20 ° C for 10 minutes, then at 0 ° C for half an hour, and then at 25 ° C for one hour. The pyridine hydrochloride was filtered under N ₂ and washed with 20 mL of THF. The filtrate was concentrated under N ₂ and then dried under high vacuum. The resulting yellow foam was taken up in 25 mL of anhydrous THF and the resulting small amount of orange-red insoluble material was filtered off under N ₂ . The filtrate is reconstituted to the above foam form.

This freshly prepared chloro compound is combined with stirring, tributylphosphine (678 mg, 3.36 mmol) in 36.5 ml 9: 1 DMF: a suspension of H ₂ 0 mixture and potassium hydrogen phosphate (294 mg, 1.69 mmol) was 55 added. The reaction mixture was stirred at 25 ° C for 35 minutes. After dilution with 120 mL EA and 60 mL brine, the layers were separated and the aqueous phase was extracted twice with EA. The combined organic layers were washed once with brine, dried over anhydrous magnesium sulfate, and quenched with N ₂ stream at 6 ° C. The concentrate was dried under high vacuum to give 12/4.

Chromatography over 100 g silica gel (height 25 cm, d = 4 cm (using _CHCl3 -at inch and 0.5% solution of CHCl ₃ in MeOH as eluent. The fractions containing the pure product 65 was collected, concentrated under a stream of N ₂ and dried under high vacuum. (12.04) of compound was obtained. impure fractions were re-chromatographed on thin plates, and 50% acetone containing silica gel) eluted with hexane. The desired uV band extraction with _CHCl3 and EA-malate further sub (12 The compound of formula (4) is obtained.

Step G (Scheme AT)

To 8.5 ml of pentane dried on anhydrous Linde 4A molecular sieve was added 0.2 ml (3.9 mol) of bromine. 1.8 ml of this 0.45 molar bromine solution are added dropwise at 0 ° C to a solution of 0.706 g (0.746 mmol) of 12/4 in a solution of (12/4) under N ₂ atmosphere. The compound of formula _{II is contained in} 18 ml of THF (freshly distilled from LAH) and 5.2 ml of Et ₂ O (dried by Linde 3A 1/16 molecular sieve). After 15 minutes, 0.42 mL (3.03 mmol) of triethylamine (molar weight = 101, density = 0.729) was added at 0 ° C and immediately

10.5 ml of ice cold DMF (distilled from calcium sulfate at 40 mm Hg and maintained above a Linde 4A molecular sieve) was added. The ice bath was removed and stirring was continued at room temperature for a further 2 hours. The reaction mixture was then poured into 3.1 ml of 1 M KH ₂ PO ₄ , 70 ml of water and 100 ml of ice-cold EA with stirring. After separation of the layers, the aqueous phase is saturated with NaCl and re-extracted with EA. The combined organic layers were washed once with brine, dried over anhydrous magnesium sulfate, and filtered. The filtrate was concentrated under a stream of N ₂ and dried under high vacuum to give crude 12/5.

This material was chromatographed on a silica gel column (60 g, 2.8 cm) in chloroform, eluting with 100 mL of 2% ethyl acetate in chloroform and 100 mL of 4% EA in chloroform, followed by chromatography on 5% EA. eluting with chloroform. The fractions containing pure compound (12/5) were collected, concentrated in a stream of N ₂ and dried under high vacuum.

Step H (Scheme AU) To a solution of anhydrous silver fluoride in mg (0.23 mmol) of 146/5 (0.15 mmol) in dry pyridine (3.5 mL) was added. The reaction mixture was stirred at room temperature for 1 hour in the dark and then poured into a solution containing 20 ml of cold water and 30 ml of EA. After separation of the layers formed, the aqueous phase was extracted twice with EA and once with chloroform. The organic phases are washed first with water and then with brine. The combined organic phases are dried over anhydrous magnesium sulfate, filtered and concentrated in a stream of N ₂ . After drying under high vacuum, 12/6 is obtained.

Purification by preparative thin layer chromatography (eluent = 40% acetone-hexane; repeated extraction of the desired u.v. band with a large amount of chloroform) afforded a slightly impure 12/6. Chromatography on silica gel, eluting with a mixture of EA and chloroform, afforded pure 12/6.

Step I (Scheme AV)

Dissolve 77 mg (0.082 mmol) of 12/6 in 0.9 ml of collidine (distilled from powdered KOH at 30 mm Hg) and pour the solution into 13.4 mg (0.1 mmol) of anhydrous Li dried under vacuum at P ₂ O _{5 for} several hours at 100 ° C. The mixture was heated in an oil bath to 120 "C under a stream of N ₂ while stirring. After heating for 30 minutes, it is cooled to 25 ° C, diluted with CH ₂ Cl ₂ and transferred to a round bottom flask for concentration in a N ₂ stream and drying under high vacuum (16181457).

The residue was partitioned between EA-water and 1M KH ₂ PO ₄ , and the aqueous phase was extracted twice with EA and once with chloroform. The organic layers were washed with brine. The combined organic phases were dried over anhydrous magnesium sulfate, filtered under a stream of N ₂ , and concentrated under high vacuum to give crude 12/7.

Purification by preparative thin layer chromatography (eluting the plate twice with 40% acetone-hexane; extracting the appropriate u.v. band again with a large amount of chloroform) afforded the recovered starting material and compound (12/7).

Step J (Scheme AX)

To a solution of 12 mg (0.07 ml) in DMSO (distilled from CaH _{2 at} 8 mm Hg and stored above Linde 4A) was added 100 liters of diisopropylamine (distilled from NaH under N ₂ ) to 49 mg (0.064 mmol). and Linde over a 4A molecular sieve). The reaction mixture was sealed for a few minutes and then allowed to stand for 2 hours. The amine and most of the DMSO were concentrated under high vacuum without external heating. The residue was rapidly passed through a silica gel chromatography column (prepared and eluted with EA) to remove residual DMSO. All the fractions having uv absorption were concentrated in a stream of N ₂ and the material was chromatographed on a thin layer of silica gel plate. (eluent = 50% EA-chloroform) a large amount of chloroform was used for repeated extraction of the desired uv band.

The recovered starting material is used twice more in the reaction and in the isolation process to provide additional desired compound.

Step K (Scheme AY)

To 5.2 g of compound 12/8 are added 0.60 ml of dioxane, 0.05 ml of ethanol, 0.35 ml of deionized water and 0.01 ml of 1 M K ₂ HPO ₄ - 35. 5 mg of 10% Pd / C was suspended in the resulting clear solution. The slurry was purged with N ₂ and then 5-6 times with 50 psi H ₂ and evacuated. Finally, shaken with 50 psi H ₂ atmosphere for 30-40 min. After centrifugation, the Pd / C is washed and centrifuged 2-3 times with 0.5 ml deionized water. The combined centrifuges were extracted with ether (5 x 1 mL). The remaining ether was removed in vacuo and the aqueous solution passed through an XAD-2 column (20 x 104 mm). 100 drops (6-7 mL) of fractions were collected under continuous UV control while sleeping with deionized water. The appearance of a strongly uv absorbent material is expected around Fraction 3-5 and usually ends around Fraction 25-30. The uv examination of the early fractions excludes the very absorbent fraction in the few 270-280s. The remaining fractions were collected and lyophilized. The residue was assayed by dissolving in 10.0 mL of deionized water and measuring the uv absorbance at 298 seconds. The assay shows a yield of 10-30% of the desired product

Example 27

Preparation of a pharmaceutical composition

A unit dose may be prepared by mixing 3- (2-60-aminoethylthio) -6-hydroxymethyl-7-oxo-1-azabicyclo (3,2,0) hept-2-ene-2-carboxylic acid with 20 mg lactose and 5 mg magnesium stearate , and the 145 mg mixture is placed in capsule # 3. Similarly, using more active ingredients and less lactose, another dosage form can be prepared in gelatin capsule # 65. Alternatively, more than 145 mg of the ingredients may be mixed in a larger capsule, tablet, or pill.

The following examples illustrate the preparation of pharmaceutical compositions:

Tablets Each tablet

3- (2-Aminoethylthio) -6-hydroxymethyl-7-oxo-1-azabicyclo (3,2,0) hept-2-ene-2-carboxylic acid 125 mg corn starch U.S.P. 6 mg

Dicalcium phosphate 192 mg lactose U.S.P. 190 mg of magnesium stearate supplemented to 800 mg

The active ingredient is mixed with dicalcium phosphate, lactose and about half of the corn starch. The mixture was then granulated with 15% corn starch pulp (6 mg) and coarsely sieved. It is dried at 45 ° C and sieved through a 16 mesh screen. The missing corn starch and magnesium stearate are added and the mixture is compressed to a tablet having a diameter of approximately 12 mm and weighing 800 mg.

Parenteral solution tablet ampoule: 3- (2-Aminoethylthio) -6-hydroxymethyl-7-oxo-1-azabicyclo (3,2,0) hept-2-ene-2-carboxylic acid 500 mg Thinner: sterile water 2 ml Ophthalmic Solution 3- (2-Aminoethylthio) -6-hydroxymethyl-7-oxo-1-azabicyclo (3,2,0) hept-2-ene-2-carboxylic acid 100 mg hydropropyl methylcellulose 5 mg sterile water 1 ml Ear solution 3- (2aminoethylthio) -6-hydroxymethyl- -7-oxo-azabicyclo (3.2.0) hept-2-ene -2-carboxylic acid 100 mg benzalkonium chloride 0.1 mg sterile water 1 ml Topical ointment 3- (2-aminoethylthio) -6-hydroxymethyl-7-oxo-1-azabicyclo (3,2,0) hept-2-ene-2-carboxylic acid 100 mg polyethylene glycol 4000 U.S.P. 400 mg polyethylene glycol 400 U.S.P. 1.0 g The active ingredient included in the above formulations alone.

or in combination with other biologically active agents such as other antibiotics such as lincomycin, penicillin, streptomycin, novobiocin, gentamycin, neomycin, colistin and kanamycin, or other therapeutic agents such as probenecid.

Example 28

The minimum inhibitory concentrations (MICs) of some of the compounds of formula I obtainable by the process of the invention are given in the following table. The serial numbers given in the table in Example 15 are used to identify the compounds.

-17181457

Serial number: MIC values (Pg / ral) 20 also 12 13 14 11 10 9 26 22 Methicillin-resistant Staphylococcus aureus > 76 25 19 7.6 17 24 7.57 34 33 12 Sthaphylococcus aureus index 0.43 0.07 0.05 0.04 0.05 0.04 0.01 0.02 0.05 0.01 Enterococcus index 15 2.4 2.69 1.77 2.69 3.69 1.61 2.8 3.02 1.5 Escherichia coli index 0.51 0.27 0.39 0.32 0.2 0.26 0.32 0.51 0.18 0.1 Enterobacter index 1.16 0.54 0.49 0.35 0.3 0.45 0.34 1.02 0.52 0.2 Klebsiella index 2.12 0.92 2 0.54 0.31 0.68 0.45 0.96 0.48 0.22 Serratia index 0.4 0.2 1.08 0.94 0.54 0.24 0.34 0.32 0.31 0.13 Indole positive and negative protein index 2.31 1.33 1.38 1.05 1.05 1.94 1.04 1.69 1.45 0.42 thienamycin resistant Pseudomonas > 70 53 34 24 37 44 20 55 51 16 Pseudomonas aeruginosa index 32 13 7.4 2.44 2.28 3.62 2.23 9.56 4.02 0.49 Methicillin-resistant Staphylococcus aureus 22 18 11 12 7.92 28 6.59 11 34 28 Sthaphylococcus aureus index 0.06 0.05 0.06 0.03 0.04 0.07 0.03 0.03 0.05 0.04 Enterococcus index 2.7 2.82 3.99 2.29 3.47 2.34 1.44 1.77 2.34 1.34 Escherichia coli index 0.34 0.13 0.23 0.09 0.21 0.19 0.2 0.43 0.25 0.19 Enterobacter index 0.6 0.24 0.45 0.15 0.32 0.29 0.22 0.42 0.39 0.22 Klebsiella index 0.63 0.39 0.59 0.32 0.51 0.51 0.41 0.67 0.67 0.36 Serratia index 0.74 0.31 0.42 0.21 0.36 0.23 0.27 0.61 0.43 0.27 Indole positive and negative protein index 1.13 1.1 1.35 0.78 U 1.1 0.96 1.67 2.2 1.18 thienamycin resistant Pseudomonas 26 36 31 31 19 46 20 28 50 50 Pseudomonas aeruginosa index 2.49 3.5 1.75 1.88 1.15 21 0.54 0.94 5.69 7

Patent claims

Claims (4)

A process for the preparation of a novel compound of formula I in the formula R @ 3 is hydroxy (1-4C) alkyl, 3 R ^{7 is} hydrogen or methyl, R ⁸ is C 1-6 alkyl optionally substituted with hydroxy, carboxy, benzyloxycarbonyl or C 2-5 alkoxycarbonyl, unsubstituted pyridyl, or 4 with the formula - (CH ₂ ) _n NQT; in the latter T is hydrogen or -C (NH) -NH ₂ or -CH = NH, Q is hydrogen or C 1-4 alkyl, n is an integer from 1 to 8, and the alkylene chain may be optionally substituted with carboxy, provided that when R ^{7 is} hydrogen and R ⁷ is 2-hydroxy, ethyl, then R ⁸ is other than 2-aminoethyl or the N-substituted derivative thereof and pharmaceutically acceptable salts thereof, characterized in that a suitable double bond is bonded between the 2 and 3 carbon atoms isomerizing, deprotecting and, if desired, forming a salt with a pharmaceutically acceptable acid or base. A process for the preparation of a compound according to claim 1, wherein R ⁸ is as defined in claim 1, with the proviso that it represents a 2-aminoethyl group or a nitrogen atom corresponding to the latter group. other than substituted derivatives, characterized in that isomerization of a suitable double bond between the 2 and 3 carbon atoms is carried out. 3. A process according to claim 1 or 2, wherein the isomerization is carried out in the presence of a base. A process for the preparation of a pharmaceutical composition, which process comprises the preparation of a compound of formula I according to claim 1, wherein R ⁷ , R ⁷ and R ⁸ are as defined in claim 1, and / or a pharmaceutically acceptable salt thereof in the manufacture of a medicament. mixed with conventional carriers and / or excipients to form a pharmaceutical composition in a known manner.