SU1424737A3 - Method of producing carbapenem derivatives - Google Patents

Abstract

The invention relates to carbopenem derivatives, in particular the preparation of compounds of the common C (0) OK, SchS-CH compound (H where R, is H, n-NO-C Hd-CHg or alkyl in the presence of a counterion; R is pyridium, or substituted pyridium, isddololium, thiomorpholine, piperazine, morpholinium, A - ethylene, unsubstituted or substituted, with antibacterial activity and used in medicine. The purpose is to simplify the process. MP) in the environment of organic solvent at a temperature of from -15 ° C to room temperature, preferably at (-15) - (15) C. As the MP, the thiol HS-AR is used, where A and R have the indicated values. A new starting thiol is an easily available compound, Allowing for Zim to conduct the process in one stage. 2 Cp. of f-ly. (U)

Description

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to

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about:

 CM

11А24737

This invention relates to methods for preparing common forpsula carbapenem derivatives.

HE

where R is from permutite S-1 C1. The column was eluted with water at a rate of 0.5 ml / min, the corresponding fdatsii were combined, and as a result of freeze-drying, a yellowish syrup of 7.0 g (100%) was obtained, which is used in the next stage.

Method B. Hydrochloride, paranitrobenzyl pyridine (4.05 g, 35 mmol) is added to the allyl protective layer of ethylene (2.1 ml, 33) in pre-cooled (ice on the bath) pyridine 10 (5.6mp. 70 mol). mmol).

n

Nzs L.,

pas if there is also a counter

RJ - pyridine or substituted pyridine, imidazolium, thiomorpholine, piperazine, morpholines;

A - ethylene, unsubstituted or substituted by methyl, cyclohexyl,

possessing antibacterial properties.

The aim of the invention is to simplify the process by using a less expensive and readily available thiol of the general formula as a starting product.

HS-A-R ,,

I where A and I have the indicated meanings.

Example 1. Preparation of 3- / 2 / 1- -thyridinium / ethylthio / -6o (1- / 1- / R / -oxo-ethyl / 7-oxo-1-azabicyclo / 3.2.0 / hept-2-e -2-carbox.

A. 1- / 2-Mercaptoethne / Pyridinium methanesulfonate.

To a suspension of pyridinium methanesulfonate in pyridine, obtained by adding dropwise methane (m

25

The mixture is heated to and moved for 75 minutes, resulting in a two-phase system. The lighter phase is removed. The remaining oil is washed with ether (5 X 10 MP) and treated with a high vacuum, resulting in the formation of the desired compound (90-100%), which is used in the next stage.

C. Para-nitrobenzyl-3- 2- / 1-pyridium / ethylthio 1-6p1- 1- / I / -oxyethyl-7-oxo-1-az abicyclo / 3.2.0 / hept-2-en-2 carboxylate chloride.

A solution of para-nitrobenzyl-6) - / I / -oxyethyl 3,7-dioxo-1-azabicyclo [3.2. O / heptane-2-carboxylate (6.09 g, 17.5 mmol) in acetonitrile (20 mp) is cooled up to the atmosphere. re nitrogen and treated successively with diisopropylethylamine (3.65 JA, 21.0 mmol) and diphenyl chlorophosphate (4.34 ml, 21.0 MMOJib). The resulting mixture was stirred for 30 minutes at 5 ° C, cooled to -5 ° C and treated successively with a solution of crude 1- / 2-mercaptoethyl / pyridinium chloride (4.3 g, 24 mmol) in

thirty

35

but) sulfonic acids (1.95 mp, 0.03 mol) Q N, N-dimethylformamide (1.0 ml) and

45

to pyridine (8.0 MP, 0.099 mol), ethylene sulfide (1.96 MP, 0.033 mol) is added with cooling. The resulting mixture is stirred at 55 ° C. for 16 hours and concentrated under reduced pressure to a thick syrup, which is mixed with a few milliliters of water. The solution is poured onto the upper part of K9lona (40 x X 16 cm) from w-bonduda C-18, which is eluted with water. Lyophilization of the corresponding fractions gives a colorless syrup of 6.5 g (91%).

B. 1- / 2-Mercaptoethyle / PyridiniumhpORID.

Method A. An aqueous solution of crude 1- / 2-mercaptoethyl / pyridinium methane sulfonate (9.4 g, 0.04 mol) is poured onto the supernatant part of the column (2.5 x 41 cm)

, - 55

drops of diisopropylethylamine (3.65 ml, 21.0 mmol). The reaction mixture was stirred at 0 ° C for 1 h, cooled to -30 ° C and stirred for another T5 min. The solid is separated by filtration and washed with cold (-30 ° C) acetonitrile (5.77 g, 65%), the filtrate and the washing liquid are combined and diluted with ether (150 ml). The upper layer is decanted, and the resin is dissolved in water (40 ml) containing a sufficient amount of acetonitrile in order to obtain a solution that is discharged onto the upper part of the column (3 x 10 cm) from C-18 / B-dapack. Then the column was eluted with a mixture of 10% acetonitrile — 90% water (150 ml) and a mixture of 50% acetonitrile — 50% water (100 ml). Matching

five

five

The mixture is heated to and moved for 75 minutes, resulting in a two-phase system. The lighter phase is removed. The remaining oil is washed with ether (5 X 10 MP) and treated with a high vacuum, resulting in the desired compound (90-100%), which is used in the next stage.

C. Para-nitrobenzyl-3- 2- / 1-pyridium / ethylthio 1-6p1- 1- / I / -oxyethyl-7-oxo-1-az abicyclo / 3.2.0 / hept-2-en-2 carboxylate chloride.

A solution of para-nitrobenzyl-6) - / I / -oxyethyl 3,7-dioxo-1-azabicyclo [3.2. O / heptane-2-carboxylate (6.09 g, 17.5 mmol) in acetonitrile (20 mp) is cooled up to the atmosphere. re nitrogen and treated successively with diisopropylethylamine (3.65 JA, 21.0 mmol) and diphenyl chlorophosphate (4.34 ml, 21.0 MMOJib). The resulting mixture was stirred for 30 minutes at 5 ° C, cooled to -5 ° C and treated successively with a solution of crude 1- / 2-mercaptoethyl / pyridinium chloride (4.3 g, 24 mmol) in

0

five

Q N, N-dimethylformamide (1.0 ml) and

45

55

drops of diisopropylethylamine (3.65 ml, 21.0 mmol). The reaction mixture was stirred at 0 ° C for 1 h, cooled to -30 ° C and stirred for another T5 min. The solid is separated by filtration and washed with cold (-30 ° C) acetonitrile (5.77 g, 65%), the filtrate and the washing liquid are combined and diluted with ether (150 ml). The upper layer is decanted, and the resin is dissolved in water (40 ml) containing a sufficient amount of acetonitrile in order to obtain a solution that is discharged onto the upper part of the column (3 x 10 cm) from C-18 / B-dapack. Then the column was eluted with a mixture of 10% acetonitrile — 90% water (150 ml) and a mixture of 50% acetonitrile — 50% water (100 ml). Matching

the fractions are combined and lyophilized after the acetonitrile has been removed under vacuum, which results in a yellowish powder. Its NMR spectrum indicates the presence of the desired compound mixed with a quantity of para-nitrobenzyl-3 / 2- / 1-pyridine / ethylthio / -6c (.- / 1 / I / -oxyethyl) -7-oxo-1-ag abicyclo / 3.2.0 / hept-2-jq-en-2-carbox shatpheny: 1.phosphate (2: 1). The powder is dissolved in water (at least) and passed through a column (1.5 x 21 cm) from permutic phosphate bu (Li monobasic sodium hydroxide with pI 7.4 (16. ml, 0.05 M) and 10% palladium on charcoal (0.16 g). The resulting mixture is subjected to hydrogenation at a pressure of 2.812 kg / cm for 1 hour at. Two the phases are separated and the organic phase is extracted with water (2 x 3 ml). The aqueous solutions are combined, washed with ether (2 x 10 ml) and poured onto the top of the column (1.5 x, 2 cm) from the | and-bondapac C-18 after separation of S -ICl with water. Lyophilization of the organic solvent mixture removes its fraction to 1, 1, g (20%) of the desired compound.

D. Para-nitrobenzyl 3- 2- / 1-pyridine-n-n / ethylthio-6o-1- / K / -oxyethyl-7- -oxo-1-asabidiclo / 3.2.0 / hept-2-2- -2-carboxyl-tetdifene blend .

The solution g1aranitrobenzyl-6 (11- 1- / K / - -oxyettz-3, 7-dioxo-1-azabicyclo- / 3.2.0 / heptan-2-carboxylate (O, 1 74 g

20

us under vacuum. Elution of the column with water after lyophilisation of the corresponding fractions gives a yellowish powder (0.062 g, 84%).

Method B. In a solution of para-nitrobenzyl-3- 2- / 1-pyr1 / ethyl-thio-but-C-1- / K / -oxyethyl-3-OXO-1-azabicyclo / 3.2.0 / hept-2- chloride en-2-carboxylate (5.77 g, 11.4 mmol) in potassium30

0.50 mmol) in acetonitrile (2 ml) of monobasic phosphate buffer is ladened to 0 ° C under nitrogen and sodium hydroxide (170 ml, 0.2 M, treated successively with diis pH 7.22) and ethyl tetrahydrofuran is added. amine (0.150 ml. O ,, 60 mmol) (30 ml), ether (30 ml) and 10% - and diphenylchlorophosphate (0} 124 ml, palladium on charcoal (5.7 g). 0.60 mmol ). The resulting solution is stirred for 30 minutes at 0 ° C and treated successively with a solution of 1- (2-mercaptoethyl) pyridium methanesulfonate (0.170 g, 0.72 mmol) in acetonitrile (0.6 ml) and diisopropane

pilsylamine (0.150 ml, 0.60 mmol) .35 days and diluted with ether. The reaction mixture is stirred at (SOO ml). The aqueous phase is separated, the resultant mixture is subjected to hydrogenation at 22 ° C at a pressure of 2.812 kg / cm for 1 hour and filtered on a pad. Gasket flush with water (2 x 15 ml). Filtrate and washing liquid so40

About C for 15 minutes, dilute with cold (0 ° C) water (7 ml) and drain the top of the column (1.5 ° x X 6.4 cm) from | and-bondapac C-18. The column is eluted with a mixture of acetonitrile (25-50%) in water (75-50%). The appropriate fractions were combined and lyophilized after the acetonitrile was removed under vacuum, resulting in a yellowish powder (0.33 g, 92%).

E. 3- 2- / 1-Pyridinium / ethno-thio- -bi, -1- / K / -oxyethyl-7 / -oxo-1-aza-bicyclo / 3.2.0 / hept-2-en-2-carboxy- lat

Method A. In a solution of para-nitrobenzyl-3- 2- / 1-pyridinium / ethylthio-6-6-1- / K / -oxyethyl-7-oxo-1-azabicyclo / 3.2.0 / hept-2-e N-carboxyl diphenylphosphate (0.16 g, 0.22 mmol) in wet tetrahydrofuran (10 ml) was added ether (10 ml), potassium ether (3 x 100 ml) and poured onto the top of the column. (4.5 X 20 cm) from a | c-bondupack C-18 after all organic solvents have been removed under vacuum. After elution of the corresponding fractions with 2.48 g (65%) of the desired compound in the form of a yellowish powder, after elution of the column with water and then with a mixture of 1% acetonitrile in water. The analytical data are identical to those performed for the compound obtained by method A.

PR and M e p 2. Preparation of 3- 2- (1- - / 3.5 dimethylpyridin-1 / ethylthio-6ot- - 1- (K) -oxyethyl-7-oxo-1-azabicyclo / 3.2.0 / hept-2-ene-2-carboxylate.

A. To a suspension of 3,5-lutyl methanesulfonate in 3,5-lutidine, obtained by adding methane (mono) sulfonic acid (0.65 ml, 0.010 mol) to cold 3,5-lutidine

50

European phosphate bu (b monobasic sodium hydroxide with pI 7.4 (16. ml, 0.05 M) and 10% palladium on charcoal (0.16 g). The resulting mixture is hydrogenated at a pressure of 2.812 kg / cm for 1 h at 2 hours.The two phases are separated and the organic phase is extracted with water (2 x 3 ml). The aqueous solutions are combined, washed with ether (2 x 10 ml) and poured onto the top of the column (1, 5 x, 2 cm) of | and-bonapac C-18 after traces of organic solvents are removed0

us under vacuum. Elution of the column with water after lyophilisation of the corresponding fractions gives a yellowish powder (0.062 g, 84%).

Method B. In a solution of para-nitrobenzyl-3- 2- / 1-pyr1 / ethyl-thio-but-C-1- / K / -oxyethyl-3-OXO-1-azabicyclo / 3.2.0 / hept-2- chloride en-2-carboxylate (5.77 g, 11.4 mmol) in tetrahydrofuran (30 ml) is added in potassium phosphate buffer monobasic sodium hydroxide (170 ml, 0.2 M, pH 7.22) ether (30 ml) and 10% palladium on charcoal (5.7 g).

tetrahydrofuran (30 ml), ether (30 ml) and 10% palladium on charcoal (5.7 g) are added to phosphate buffer monobasic sodium hydroxide (170 ml, 0.2 M, pH 7.22). .

day and dilute with ether (OO ml). The aqueous phase is separated, the resultant mixture is subjected to hydrogenation at 22 ° C at a pressure of 2.812 kg / cm for 1 h and filtered on a pad. Gasket flush with water (2 x 15 ml). Filtrate and washing liquid so0

five

washed with ether (3 x 100 ml) and poured onto the upper part of the column (4.5 x 20 cm) from c-18 bondapac after all organic solvents have been removed under vacuum. Elution of the column with water and then with a mixture of 1% acetonitrile in water, after lyophilization of the corresponding fractions, 2.48 g (65%) of the desired compound as a yellowish powder. The analytical data are identical to those performed for the compound obtained by method A.

PR and M e p 2. Preparation of 3- 2- (1- - / 3.5 dimethylpyridin-1 / ethylthio-6ot- - 1- (K) -oxyethyl-7-oxo-1-azabicyclo / 3.2.0 / hept-2-ene-2-carboxylate.

A. To a suspension of 3,5-lutyl methanesulfonate in 3,5-lutidine, obtained by adding methane (mono) sulfonic acid (0.65 ml, 0.010 mol) to cold 3,5-lutidine

0

(2.51 ml, 0.022 mol) ethylene sulfide (0.655 ml, 0.011 mol) was added. The resulting mixture was stirred under a nitrogen atmosphere at 55 ° C for 2 hours, cooled to 23 ° C and diluted with water (5 ml) and ether (5 mp). The organic layer is separated and the aqueous solution is washed.

-7-oxo-1-azabicyclo / 3.2.0 / hept-2-en-2-carboxylate.

Into a solution of para-nitrobenzl-3- 2- - / 1- / 3,5-dimethylpyridinium / ethyl-Thio - -boL-1- / K / -oxyethyl-3-oxo-1-azabicyclo / 3.2.0 / hept-2- ene-2-carboxylate of diphenyl phosphate (0.600 g, 0.80 mmol) in wet tetrahydrofuran (36 mp)

ether (6x4 nl). Traces of pro-u-add ether- (36 ml), ca15

20

of the ester is removed under vacuum, and the solution is applied to the top of the column (2.5 x 6.0 cm) from (U-Bondapak C-18. The column is eluted with water, freeze-drying the corresponding fractions to obtain a colorless syrup (2 , And g, 91%).

Calculated: C, 44.09; H, 6.66; N 5.14; S 23.54.

C, oHnNO S; i-0,5H20

Found: C, 44.26; H 6.49; N 5.17; S 2A, 18.

B. Paranitrobenzyl 3- 2- / 1- / 3,5- -dimethylpyridine // - ethyl-d6oi. - / K / -oxyethyl-7-oxo-1-azabicyclo (3.2.0/hept-2-ene-2-carboxylate diphenyl phosphate).

In a cold (o C) solution of para-nitro-benzyl .- / 1- / I / -oxyethyl / -3, 7-dioxo-1-azabicyclo / 3.2.0 / heptane-2-carg jg of boxyl (0.523 g, 1.50 mmol) in acetonitrile (6.0 ml), which is maintained under a nitrogen atmosphere, diisopropylethylamine (0.314 ml, 1.8 mmol) is added, followed by diphenyl chlorophosphate (0.373 ml, 1.8 mmol). The reaction mixture is stirred for 30 minutes and treated with a solution of 1- / 2-mercaptoethyl- / 3,5-dimethyl-pyridine methanesulphonate (0.493 g, 1.87 mmol) in acetonitrile (1.9 ml), and then diisopropylethylamine (0.314 mp, 1.8 mmol). The reaction mixture was stirred at 0 ° C for 1 h, diluted with cold (0 ° C) water (26 ml) and poured onto the top of a column (7.0 x 3.5 cm) from jU-bonnapack C-18. Elution of the column with a mixture of 25–50% acetonitrile — 75–50% water after lyophilization of the corresponding fractions gives 1.01 g (90%)

lithium phosphate buffer monobasic sodium hydroxide (0.05 M, pH 7.4, 44 ml) and 10% palladium on charcoal (0.60 g). The resulting mixture is then subjected to hydrogenation at a pressure of 2.812 kg / cm and a temperature of 28 seconds for 1.25 hours. The organic layer is separated and extracted with buffer (2 X 5 nL). The aqueous layers were combined, filtered through a Celite pad, washed with ether (40 ml), treated under vacuum to remove traces of organic solvents, and poured onto the top 25 of the column (2.5 x 10.0 cm) from | -bondupac C-18. Elution of the column with water and lyophilization of the appropriate fractions gave the title compound (0.186 g, 64%) as a yellowish powder.

Example 3. Preparation of (5R, 6S) - -3-2- / 3-hydroxymethylpyridino (eth) 1 - -TIO / -6- 1- (K) -oxyethyl-7-OXO-1-azabicyclo / 3.2.0 / hept -2-ene-2-carboxylate.

A. 3-Oxymethyl-1/2-mercato-ethyl / pyridine trifluoromethanesulfonate.

Trifluoromethane (mono) sulfonic acid (1.327 ml, 0.015 mol) was added dropwise to 3-pyridine methanol (2.91 ml, 0.030 mol), and then ethylene sulfide (0.89 ml, 0.015 mol) was added to the mixture. The resulting mixture is heated (oil / bath) to 50-70 ° C under nitrogen for 20 hours. The reaction mixture is then transferred to (15 ml) and extracted with (5 x 5 ml). The aqueous phase is concentrated under vacuum, and then applied in a inverted column.

the desired compound as yellow phase chromatography C, j. Eluirovago powder.

Calculated: C 58.03; H 5.26; N 5, A8; S 4.18.

,, o SP-H, 0

Found: C 57.98; H 5.05; N 5.22; 55 get almost colorless oil. After drying under vacuum (4,34.le), () it is DC. 3- 2- / 1- / 3,5-Dimethylpyridinium with water, and then evaporating the corresponding fractions gives a gray-yellow oil. This material is again subjected to chromatography, with the result that

nium // ethclthio-6c (.- 1- / E / -oxyethyl) the desired product (4.50 g, 94%) as a viscous oil.

-7-oxo-1-azabicyclo / 3.2.0 / hept-2-en-2-carboxylate.

Into a solution of para-nitrobenzl-3- 2- - / 1- / 3,5-dimethylpyridinium / ethyl-Thio - -boL-1- / K / -oxyethyl-3-oxo-1-azabicyclo / 3.2.0 / hept-2- ene-2-carboxylate of diphenyl phosphate (0.600 g, 0.80 mmol) in wet tetrahydrofuran (36 mp)

five

0

g.

lithium phosphate buffer monobasic sodium hydroxide (0.05 M, pH 7.4, 44 ml) and 10% palladium on charcoal (0.60 g). The resulting mixture is then subjected to hydrogenation at a pressure of 2.812 kg / cm and a temperature of 28 seconds for 1.25 hours. The organic layer is separated and extracted with buffer (2 X 5 nL). The aqueous layers were combined, filtered through a Celite pad, washed with ether (40 ml), treated under vacuum to remove traces of organic solvents, and poured onto the top 5 of the column (2.5 x 10.0 cm) from | -bondupac C-18. Elution of the column with water and lyophilization of the appropriate fractions gave the title compound (0.186 g, 64%) as a yellowish powder.

Example 3. Preparation of (5R, 6S) - -3-2- / 3-hydroxymethylpyridino (eth) 1 - -TIO / -6- 1- (K) -oxyethyl-7-OXO-1-azabicyclo / 3.2.0 / hept -2-ene-2-carboxylate.

A. 3-Oxymethyl-1/2-mercato-ethyl / pyridine trifluoromethanesulfonate.

Trifluoromethane (mono) sulfonic acid (1.327 ml, 0.015 mol) was added dropwise to 3-pyridine methanol (2.91 ml, 0.030 mol), and then ethylene sulfide (0.89 ml, 0.015 mol) was added to the mixture. The resulting mixture is heated (oil / bath) to 50-70 ° C under nitrogen for 20 hours. The reaction mixture is then transferred to (15 ml) and extracted with (5 x 5 ml). The aqueous phase is concentrated under vacuum, and then applied on a reversed column.

water, and then evaporation of the corresponding fractions gives a gray-yellow oil. This material is again subjected to chromatography, with the result that

The desired product (4.50 g, 94%) is in the form of a viscous oil.

B.Paranitrobenz1SH- (5K, 68- / 3-C2- / Z-hydroxymethylpyridinio ethylthio / -6 -1 -1- / K / -oxyethyl-7-ok co-1-azabicyclo / 3.2.0 / hept -2-en-2-carboxylate.

B solution of para-nitrobenzyl- / 5K, b5 / -6- 1- / K / -oxyethyl-3,7-dioxo--1-azabicyclo / H, 2.0 / heptane-2-carboxylate (0.174 g, 0, 50 mmol) in 2 ml of dry acetonitrile diisopropylethylamine (0.096 ml, 0.55 mmol) is added at 0 ° C under nitrogen atmosphere. Diphenylchlorophosphate (0.114 MP, 0.55 mmol) is then added dropwise and the reaction mixture is stirred at-0 ° C for 30 minutes. A solution of 3-oxymethyl-1- / 2-mercaptoethyl / pyridinium trifluoromethanesulfonate (0.223 g, 0.70 mmol) in 0.50 MP of acetonitrile and diisopropylethylamine (0.122 ml,

0.70 mmol). After being left at 0 ° C for 30 minutes, the reaction mixture was concentrated under vacuum and the resulting yellow resin was transferred to water (a sufficient amount of acetonitrile was added to accelerate the dissolution of the resin). This solution was applied to the column by reverse phase chromatography. C-18 which is then eluted with a mixture of 15% acetonitrile. As a result of the lyophilization of the corresponding fractions, the product is obtained (0.305 g, 81%) as a beige solid.

C. (5K, 68) -3- 2- / 3-hydroxymethyl-1-pyrio / ethylthio-1-6- | 1- / K / -oxyeth-1 -7-oxo-1-az abicyclo / 3.2.0 / hept-2- e-2-carboxylate.

Into the solution of paranitrobenzyl 5R, 6S) - -3- / 2- / 3-hydroxymethylpyridinio / methylthio / - -6- / 1- / E / -oxistil / -7-oxo-1-azabicyclo / 3.2.O / hept -2-en-2-carboxylate of diphenyl phosphate (0.145 g, 0.194 mmol) in 10 MP of THF containing 5 drops of water was added 6.0 MP of phosphate buffer (0.05 M, pH 7.4), 0.145 g of 10% - palladium on charcoal and 10 ml of ether. The mixture is subjected to hydrogenation (Parr) at a pressure of 2.812 kg / cm for 1 h, and then filtered through a Celite pad. Filter cake is washed with a small amount of water and ether, and then the aqueous phase is separated and extracted with ether. The aqueous solution is then cooled to 0 ° C., a pH of 7.0 is provided with a buffer of pH 7.4. After removal of residual volatile materials under vacuum, the aqueous

five

0 5

0

the solution was applied to a C-18 C phase chromatography column, which was then eluted with water. As a result of freeze-drying of the appropriate fractions, the product (36 mg, 51%) is obtained in a light yellow solid saw. Purification using high-pressure reverse phase liquid chromatography affords the pure product (31 mg, 41%) as a solid.

PRI me R 4. Obtaining (5R, 6S) - -Z-- 2- / 4-hydroxymethylpyrillinio / ethylthio-6-1- / I / -oxyethyl-7-oxo-1-azabicyclo / 3.2.0 / hept-2-en-2-carboxylate.

A.4-Oxymethyl-1- / 2-merkaito i and I / - pufidinium trifluoromethanesulfoate.

Trifluoromethane (mono) sulfonic acid (1.327 ml, 0.015 mol) was added dropwise to a solution of 4-pyridinemethanol (1.635 g, 0.015 mol) in 10 ml at 0 s under a nitrogen atmosphere. The tan oil is quickly separated. To this mixture is added an additional equivalent of 4-pyr1 of methanol (1.635 g, 0.015 mol). And the solvent is removed with a diluted dhl of H1X, resulting in an oil. Ethylene sulfate (0.891 ml, 0.015 mol) is added to this oil and the resulting homogeneous mixture is heated (oil per bath) to approximately 60 ° C for 3 hours. Then the reaction mixture is transferred to 15 ml of water and aqueous solution washed with (5x5 ml). After removing the residual organic solvent under vacuum, the aqueous solution is poured onto a C-1 column. 8. Elution with water and subsequent evaporation of the appropriate fractions gives an oil, which is then dried under vacuum over P — O the required Recreatives Products (4.64 g, 97%) as a colorless oil.

B, Para-nitrobenzyl 5R, - / 4-hydroxymethylpyridinio / ethylthio-6-1 / E / oxyethyl / -7-oxo-1-azabicyclo 3.2.o hept-2-en-carboxylate diphenyl phosphate.

Into a solution of para-nitrobenyl / 5R, 6S / -6- 1- (K) -oxyethyl / -3,7-dioxo--1-azabicyclo / 3.2.0 / heptane-2-carboxylate (0.348 g, 1.0 mmol ) in 5 ml

dry acetohydrnol is added dropwise lyisogrogrotylethylamine (0.191 ml, 1.1 mmol) under nitrogen and then diphenyl chlorophosphate (0.228 ml, 1.1%). The resulting yellowish-golden solution was stirred at 0 ° C for AO minutes. A-hydroxymethyl-1- [2-mercaptoethyl / pyridium trifluoromethanesulfopate solution (0, A47 g, 1.4 mmol) in 1 ml was added to this solution. acetonitrile, and then diisopropylethylamine (0.191 ml, 1.1 mmol). The reddish-black resin is separated from the reaction mixture. After over 20 minutes at 0 ° C, the reaction mixture is filtered and concentrated under vacuum. The residue is transferred to a minimum volume of acetonitrile-water (1: 1) and then applied to a C-18 reverse phase chromatography column. Elution with a 25% acetone gril-water mixture and subsequent lyophilization of the appropriate fractions gave the product (0.353 g, 47%) as a cream-colored solid.

C, / 5H, 65 / -3- 2- / 4-Oxymethylpyridinium / ethylthio-6- 1- / K / -oxy-H-7-oxo-1 azabicyclo / 3.2.0 / hept 2-en-2 carboxylate

The mixture of paranitrobenzyl / 5R, 6S / - 2- / 4-hydroxymethylpyridinio / ethylthio - -6-1- / K / -oxyethyl-7-oxo-1-azabicyclo / 3.2.0 / hept-2-en-2- carboxylate diphenylphosphate (0.348 g, 0.465 mmol) and 10% palladium on charcoal (0.35 g) in 11 ml of phosphate buffer (0.05 M, pH 7.4), 5 MP THF and 10 ml of simple efgf hydrogenated at a pressure of 2.812 kg / cm for 1.25 hours. The mixture is then filtered through a Celite pad, a. the aqueous phase is washed with ether (3X). The pH of the aqueous solution is then adjusted to 7.0 with an additional amount of buffer with a pH of 7.4. After removing the residual volatile materials under vacuum, the aqueous solution is applied to a C-18 reverse phase chromatography column. A yellow-brown solid is obtained by elution with a mixture of 2% acetonitrile-water and subsequent freeze-drying. This material is subjected to chromatography (C-18, reversed phase / H O), resulting in the desired product (0.060 g, 36% ) in the form of a light yellow solid.

one

PRI me R 5. Obtaining 3- 2- / 1- - / 2-methylpyrillinium / ethylthio-6 - 1- - / K / -oxyethyl-7-oxo-1-azabicyclo- / 3.2 .0 / hept- 2-en-carboxes1.

A.1- / 2-Mercaptoethyl / -2-methylpyridine methanesulfonate.

To a suspension of 2-methylpyridinium methanesulfonate in 2-methylpyridine, obtained by adding methane (mono) sulfonic acid (0.65 ml, 0.050 mol) to cold 2-methylpyridine (2.17 MP, 0.022 mol), add sulfide ethylene (0.655 ml, 0.011 0.011 mol). The reaction mixture is stirred under a nitrogen atmosphere at 55 ° C for 21 hours, cooled to 23 ° C and diluted with water (5 ml). The aqueous solution is washed with ether (6 x X 4 ml), treated under vacuum to remove traces of organic solvents, and poured onto the top of the column (2.5 x 10.0 cm) from c-18 bondapac. The column was eluted with water, and the corresponding fractions were lyophilized, resulting in 2.13 g (85%) of the title compound.

B. Para-trobenzyl-3- 2- (1- / 2-methylpyridinium / ethylthio-6O1-1- / K / -oxy-7-oxo-1-azabicyclo 0 / 3.2.0 / hept-2-ene- 2-carboxylate. Diphenyl phosphate.

In a cold (0 ° C) solution of paranitrobenzsht-boS-1- (K) -oxyethyl-3,7-dioxo-1-azabicyclo / 3.2. O / heptane-2-carboxylate (0.523 g, 1 , 50 mmol) in acetonitrile (6 ml), which is maintained under a nitrogen atmosphere, diisopropylethylamine (0.314 ml, 1.80 mmol) is added, and then diphenyl chlorophosphate (0.373 ml, 1.80 mmol) is added. The reaction mixture is stirred for 30 minutes at 0 ° C and treated with a solution of 1- / 2-mercaptoethyl / -2-methylpyridine-methane sulfonate (0.530 g, 2.16 mmol) in acetonitrile (18 mp), and then the diisopne ethyl amine (0.314 ml, 1.8 mmol) The reaction mixture was stirred at 0 ° C for 1 h, diluted with cold () water (26 mp) and poured onto the top of the column (3.5 x 7.0 cm) from jU-bondapac C-18. As a result of elution of the column with 25% acetonitrile — 75% water and a 50% mixture.

acetonitric - 50% water is obtained after freeze-drying the corresponding fractions of 1.06 g (96%) of the title compound as a yellowish powder,

P. 3- 2- / 1- / 2-Methylpyridinium // - / ethylthioz-6oi- 1- / K / -oxyethyl-3-oxy-1-azabacyclo / 3.2.0 / hept-2-en-2 - carboxylate.

,

Into the solution of paranitrobenzyl-3- 2- - / 1- / 2-methylpyridinium // ethylthio -LoC- - 1- / K / -oxyeth-1 -7-oxo-1-azabicyclo / 3.2.0 / hept-2-ene -2-carboxylate diphenyphosphate (0.66 g, 0.90 mmol) in wet tetrahydrofuran (34 ml) was added ether (34 ml), potassium phosphate buffer monobasic sodium hydroxide (0.15 M, 16.5 ml, pH 7.22) and 10% palladium on charcoal (0.66 g). The resulting mixture is hydrogenated at a pressure of 2.812 kg / cm for 1.25 hours. The organic layer is separated and extracted with buffer (2 x 6 ml). The aqueous layers were combined, filtered through a Celite pad, washed with ether (40 ml), treated with noh vacuum to remove traces of organic solvents, and poured onto the top of the column (2.5 x 10 cm) | and Bondapak C -18. By elution of the column with water and freeze-drying of the appropriate fractions, the desired compound (0.098 g, 31%) was obtained as a yellowish powder. EXAMPLE 6 Preparation of 3- 2- - / 1- / 4-methylpyridinium / ethylthio-6oi- - l- / K / -oxyethyl-7-oxo-1-azabicyclo / 3.2.0 / hept -2-en-2-carboxylate.

A. 1- / 2-Mercaptoethyl / -4-methyl-pyridinium methanesulfonate.

To a suspension of 4-picolinium methanesulfonate in 4-picoline, obtained by adding methane (mono) sulfonic acid (0.65 ml, 0.010 mol) to A-picoline (2.14 ml, 0.022 mol), while cooling ethylene sulfide (0.655 MP, 0.011 mol). The reaction mixture was stirred under nitrogen at 55 ° C for 24 hours, cooled to 23 ° C and diluted with water (5 ml) and ether (10 ml). The organic layer is separated, and the aqueous layer is washed with ether (5x5 ml) and applied to the top of the column (2.5 x X 10 cm) from | U-Bondapac C-18 after it has been removed.

5 o Q

five

0

Traces of ether have been removed at nominal pressure. As a result of blending with a mixture of 15% acetonitrile — 85% water, the columns are obtained by removing the lyophilis from the corresponding fractions, a colorless syrup (2.66 g, 100%).

B, 1- / 2-Mercapt6ethyl / -4-methylpyridinium and para-toluenesulfonate.

In a suspension of para-toluene (mono) sulfonic acid (1.72 g, O, O G mol) p benzene (6.5 ml) A-picoline (1.17 ml, 0.012 mol) is added ;. The resulting mixture was stirred under nitrogen at 23 ° C for 30 minutes, treated with ethylene sulfide (0.65 ml, 0.011 mol) and stirred at 75 ° C for 24 hours. Ethyl sulfide (0, 65 ml, 0.011 mol) and stirring is continued for another 24 hours at 75 ° C. The reaction mixture is cooled to 23 ° C and diluted with water (5 ml) and ether (8 ml). The aqueous layer was separated and washed with ether (3x8 ml). Traces of organic solvents are removed in vacuo, and the compound is chromatographed on a j4 C-18 monoconducer using water as an eluting solvent, to give 2.94 g (90%) of the desired compound as a colorless syrup.

C.Paranitrobenzyl-3-C2-1- / 4-methylpyridinium // ethylthioZ-6o (.- N / K-hydroxyethyl-7-ca co-1-az abicicle o / 3.2.0 / - hept 2-ene-2-carboxylate diphenyl phosphate.

In cold (О С) solution of para-nitro-benzyl-be- (1- / K) -oxyethyl / 3,7-diox-co-1-azabicyclo / S.1.0 / 2-carboxylate heptin (0.522 g, 1, 5) in acetonitrile (6 ml), maintained under a nitrogen atmosphere, diisopropylamine (0.314 ml, 1.8 mmol) is added, followed by diphenyl chlorophosphate (0.373 ml, 1.9 mmol). The reaction mixture is stirred for 45 minutes and treated dropwise with a solution of 1- / 2-mercaptoethne / -4-methylpyridine methanesulfonate (0.539 g, 2.16 mmol) in acetonitrile (1.8 ml), and then diisopropylethylamine (0.314 ml, 1.8 mmol). The reaction mixture was stirred at 0 ° C for 1 hour, diluted with cold (0 ° C) water (24 ml) and poured onto the top of the column (2.5 x 8.5 cm) e (I-Bondapack C-18. As a result of elution columns first with a mixture of 25% acetonitrile - 75% water (100 ml), then with a mixture of 50% acetonitrile - 50%

water (100 ml) n as a result of lnoflization of the corresponding fractions, 0.91 g (83%) of the desired compound is obtained as a yellowish powder,

 Calculated; C 56.84, And 5.17; N 5.52; S 4.21.

C36H, 6N, 0, oSP.1, Found: C 56.89 and 5.13, N 5.19; S 4.41.

D. 3- 2- / 1- / 4-Methylpyridinium // - / ethylthio-6o.- 1- / K / -oxy-tylZ-7-oxy-1-azabicyclo0 / 3.2.0 / hept-2-ene -2- carboxylate.

To the solution of paranitrobenzyl - / 1- / 4-methylpyridinium-ethylthio-6 {/ - - 1- / K / -oxyethyl-7-oxo-1-azabishpslo / 3.2.0 / hept-2-en-2-carboxylate - phenyl phosphate (0.587 g, 0.80 mmol), ether (30 ml) is added to wet tetrahydrofuran (30 ml), potassium phosphate buffer monobasic sodium hydroxide (0.15 M, 14.7 ml pH 7.22) and 10% palladium on charcoal (0.59 g). The resulting mixture was subjected to hydrogenation at a pressure of 2.812 kg / cm and temperature for 1.25 hours. The organic layer was separated and extracted with buffer (2 x 6 ml). The aqueous extracts are combined, filtered through a Celite pad, washed with ether (3 X 20 ml), treated under vacuum to remove traces of organic solvents, and poured into the upper part of the column (2.5 x

X 10 cm) from | and -20-C-18. By elution with water and freeze-drying of the appropriate fractions, 0.136 g (49%) of the desired compound is obtained. In the form of a yellowish powder.

Example 7. Preparation of / 5R / 3- - / 2- / 4-Methylthiopyridinio / ethylthio / - - / 68 / - // 1Д / -ooxyethanol / -7-oxr-1-azabicycl / 3.2.0 / hept-2 en-2-carboxylate

A. 4-methylthiopyridine.

4-mercaptopyridine (5.55 g, 50.0 mmol, Eldrich) is dissolved in boiling absolute E tOH (50 ml). Insoluble material was removed by filtration through Celite. The filtrate is heated to re-dissolve, and when it is heated to 50 ° C, meth-thiodide (3.17 ml, 51.0 mmol, Eldrich) is added in one portion. The mixture is cooled to crystallize. Filtration of a solid gives 6.77 g

0

c 0 5 p

..

five

(26.7 mmol, yield 53.5%) of the title compound as iodohydrate.

The iodohydrate (6.33 g, 25.0 mmol) was dissolved in H-jO (40 ml), and the insoluble material was removed and washed with water (10 ml). To the filtrate is added with a tablet of NaOH (5 g) and extracted with (3 x 25 ml), saturating the aqueous layer with sodium chloride. The combined organic extracts were washed with brine (x 2), dried (MgSO4) and quantified, resulting in 2.92 g (23.4 mmol, total yield 50%) of the title compound as an oil.

B.4-Methylthio-N- / 2-mercaptoethyl / - pyridine methanesulfonate.

4-methylthiopyridine (2.75 g, 22.0 mmol) is slowly added to methanesulfonic acid (0.65 ml, 10.5 mmol) while cooling in an ice bath. Ethylene sulfide (0.66 ml, 11.0 mmol, Eldrich) is added to this solution and the mixture is heated to a temperature of 50-60 ° C, which is maintained for 21 hours. During the reaction, the solid goes into solution. After cooling, the reaction mixture was dissolved in water (5 mp) and washed (5x4 ml). The cloudy aqueous layer is filtered through Celite, and the filtrate is purified using a silica gel chromatographic column (C-18 u-bonepac, 10 g), which is water-cooled. Collect fractions of 10 MP. Fractions 2 and 3 are combined and purified again using a reverse phase chromatography column. Fraction 2 yielded 1.258 g (2.48 mmol, yield 42.6%) of the desired compound. In the form of a viscous oil.

Methanesulfonic acid and ethylene sulfide are preliminarily distilled.

C. Chloride / 5K / -paranitrobenzyl-3-C2- / 4-methylthiopyridine / -ethylthio7- - / 65 / - / 1К / -oxyethyl-7-oxo-1-aza-bicyclo / 3.2.0 / hept-2 en-2-carboxylate

To solution / 5К / -paranitrobenzyl-3; 7-dioxo- / 65 / - / 1К / -oxyethyl -1-azabicyclo / 3.2.0 / heptane- / 2К / -carboxylate (475 mg, 1.36 mmol) and diisopropylethylamine (0.24 ml, 1.4 mmol) in C} CUM (5 ml) was added at 0-5 ° C azotiphenyl chlorophosphate (0.29 ml, 1.41 mmol). The mixture is stirred at 0-5 ° C for 30 minutes. Into this mixture

1 51

by adding a ktg oily suspension of 4-methylthio-N- / 2-mercaptoethyl / pyridinium methanesulfonate (678 mg, 1.45 mmod, purity 60%) in CHjCN (1.5 ml), and then diisopropylethylamine (0.24 un, 1.4 mmol). The mixture was stirred at 0-5 ° C for 1 hour. Immediately after the addition of the base, a yellowish precipitate formed. The precipitate was isolated by filtration and washed with cold CHjCN (3 ml), resulting in 413 mg of a yellowish solid. It is triturated with 10% MeOH in water (5 ml) in order to obtain 341 mg (0.618 mmol, 45.4% yield) of the title compound as white crystals, m.p., 118-120 ° C:

Calculated: C, 50.56; H 4.95, N 7.37.

, 04, S ,, Cl-H, 0

Found: C 50.63; H 4.72, N 6.89

D. / 5R / -3- 2- / 4-Metsh1Toyoyiridino / ethylthio / - / 68 / - / 1К / -oxyethes {-7 -oxo-1-azabicyclo / 3.2.0 / hept-2-e- - 2-caroxylate.

B.Paranitrobenzyl-3- 2- / 3-methoxy-1-pyridinium chloride / methylthio 6uL- / l- / P / -oxyethyl-7-oxo-1-azabicyclo / 3.2.0 / hept-2-ene -2-carboxylate.

Cold (OH) solution of paranitrobenzyl-6oi- / 1 - / K / -oxyethyl / -3,7-dioxo-1-azabicyclo / 3.2.0 / heptane-2-carboxylate (1.04 g , 3 mmol) in acetonitrile (12 ml) is treated in drops / 5K / -paranitrobenzyl-3- / 2- / 4-methylthiopyridinio / ethylthio / - / 6S / - / 1R / -oxyethyl-7-OXO-1- azabicyclo / 3.

hept-2-ene-2-carboxylate chloride (380 mg,

0.688 mmol) is dissolved in THF

(31.5 MP) and phosphate solution

(31.5 ml) and phosphate buffer with pH

7.40 (31.5 ml, 0.05 MFisher) and dilution-. , olute with EtjO (31.5 ml). This l m diisopropylstilamine (0.63 ml,

the solution is mixed with 10% Pd / C (3.6 mmol) and diphenyl chlorophosphate

(380 mg, Engelhardt) and subjected (0.75 MP, 3.6 mmol) and perepeptvayut

at 0 With in 30 minutes The resulting ethanol phosphate is treated with 1-2- / mercaptoztil / -3-methoxylthiridium methanesulfonate (1.14 g, 4.30 mmol) in (7 ml), diisoprohydrogenation at a pressure of 2p461 kg / cm on a Parr vibrator at room temperature in for 1 hour. The aqueous layer is filtered through Celite to remove the catalyst, and then the Celite gasket is drawn with water (2 x 5 ml). The filtrate and the washing liquid are combined and washed with the aid of (2 X 30 ml). The aqueous layer is treated under vacuum to remove organic solvents and purified on a reverse phase chromatographic column (C-18 (U-Bondapac, 13 g, Waters Associates), which is eluted with water. Fractions containing absorption in the UV spectrum in range 307 im so40

pilyethylamine (0.63 ml, 4.30 mmol), stirred for 30 minutes and cooled to a temperature of -10 ° C, which is maintained for 30 minutes. The solid that precipitates from the mixture is filtered, washed with cold acetonitrile (2 ml) and dried, and the resultant compound is obtained (1.32 g, 82% yield).

C, (3-Methoxy-1-pyr-Schin1-sh / - etipio-6ot- / 1 - / K / hydroxyethyl-7-ox- (-1-azabicyclo / 3.2.0 / hept-2-en-2b (about 1 l) and as a result 55-carboxylate.

127 mg of a solution of para-nitrobenzyl-3 2- / 3-me (0.334 mmol, yield: 48.5%) of iso-homoxy-1-pyridnium xdo id / ethylthio-6 "" g / 1 - / K / -oxyethyl 1-7-oxo- 1-azode compounds as a yellow powder.

sixteen

0

In the example, the preparation of (3-methoxy-1-pyridine / ethylthio - - / K) -oxyoeth-7-oxo-1-az-bicyclo / 3.2.0 / hept-2-en-2-carboxyliclate.

A.1- / 2-Mercaptoethyl / 3-methoxy-pyridinium methanesulfonate.

In pre-cooled (5 C) methoxypyridine (698 mg, 6.4 mmol) is added dropwise methane (mono) sul1) - phyloacid (0.216 ml, 3.05 mmol) and ethylene sulfide (O, 19 ml, 3, 2 mmol). The mixture is then heated to and kept at this temperature for 18 hours, cooled to 20 ° C, diluted with water (10 ml) and npoMi-housed with ether (3 x 10 ml). I process the aqueous phase) under a deep vacuum unit for 15 min, and then it is poured onto a C-18 reversed-phase chromatographic column. The desired compound is eluted with water. The corresponding fractions were combined and pressed into vacuum under a deep vacuum, whereby the desired thiol was obtained (61.6 mg, yield 76.3%).

B.Paranitrobenzyl-3- 2- / 3-methoxy-1-pyridinium chloride / methylthio 6uL- / l- / P / -oxyethyl-7-oxo-1-azabicyclo / 3.2.0 / hept-2-ene -2-carboxylate.

Cold (OH) solution of paranitrobenzyl-6oi- / 1 - / K / -oxyethyl / -3,7-dioxo-1-azabicyclo / 3.2.0 / heptane-2-carboxylate (1.04 g , 3 mmol) in acetonitrile (12 ml) is treated with a cap20

25

0

. , l m diisopropylstilamine (0.63 ml,

40

at 0 With in 30 minutes The resulting ethanol phosphate is treated with 1-2- / mercaptoztil / -3-methoxyl thyridine methanesulfonate (1.14 g, 4.30 mmol) in (7 ml), diisopropylethylamine (0.63 ml, 4.30 mmol), stirred 30 min and cooled to a temperature of -10 ° C, which is maintained for 30 minutes The solid that precipitates from the mixture is filtered, washed with cold acetonitrile (2 ml) and dried, and the resultant compound is obtained (1.32 g, 82% yield).

C, (3-Methoxy-1-pyr-Schin1-sh / - etipio-6ot- / 1 - / K / hydroxyethyl-7-ox-c (-1-azabicyclo / 3.2.0 / hept-2-en-2 toxy-1-pyridniumhd id / -ethylthio-6 "" g / 1 - / K / -oxyethyl 1-7-oxo-1-azabicyclo / 3 .2.0 / hept 2-ene-2-carboxylate (600 mg, - 1.12 mmol ) in THF (25 ml of ether (25 ml) and phosphate buffer pH 7.4 (0.1 M, 25 ml) is subjected to hydrogenation on a Parr vibrator using Pd / C (1.1 g) for 1 h at pressure of 2.812 kg / cm. The mixture is diluted with ether and the aqueous phase is filtered through filter paper.

Water layer npONfblBaiOT simple EF1-FOM

(2 X 20 ml), are treated under vacuum and poured onto a reverse phase chromatographic column on silica gel. The desired compound is eluted with water containing 2 and 5% acetonitrile. The corresponding fractions are combined and lyophilized, resulting in a yellow solid that is again purified by POM1OC high pressure chromatography, resulting in a penemcarboxyl (150 mg, 38%).

PRI and e R 9. Getting / 5Rj6S / - 3- 2- / 3-Mstilthiopyridine / ethylthio - -6-G1- / K / -oxyethyl-7-oxo-1-azabi

cyclo L3.2. OJ hept-2-en-2-carbox1shata.

A, 3-Methylthio-1- / 2-mercaptoethyl / - pyridinium chloride.

To a solution of 3-methylthiopyridine (2.00 g, 0.016 mol) in 10 MP ether is added 15 ml of IN solution of HCl and the mixture is placed on a vibrator. The aqueous phase is separated, washed with 10 ml of ether and then added. The hydrochloride is then dried under vacuum (), and a white solid is obtained. 3-methylthiopyridine (1.88 g, 0.015 mol, and ethylene sulfide (0.89 ml, 0.015 mol, and the resulting mixture is heated (oil per bath) to a temperature of 55-65 ° C) is added to this solid hydrochloride salt. in a nitrogen atmosphere that is maintained for 15 hours. A slightly turbid oily liquid is obtained, which is transferred to 125 ml of water, then it is washed with the aid. The aqueous solution is concentrated to approximately 25 ml, and then a few drops of acetonitrile are added for to make the mixture homogeneous. The resulting aqueous solution is applied on a raschen- but-faeovuyu chromatographic column C-18. Elution with water and subsequent evaporation sootvetstvuyuprta

about

5 0 5

Q

,

five

fractions gives the desired product (2.66 g, 80%) as a gray-yellow viscous oil.

B. Para-nitrobenzyl / 5K, 65 / -3- 2- - / 3-methylthiopyridine / ethylthioZ-6- 1- - / K / -oxyethane-7-oxo-1-azabicyclo 3.2.6 hept-2-ene- 2-carboxylate.

A solution of para-nitrobenzyl / 5H, B5 / - -6- I- / R / -oxethyl-3,7-DIOXO-1-aza-bicyclo / 3.2.0 / heptane-2-carboxylate (0.522 g, 1.50 mmol) in 7 ml of dry acetonitrile are cooled to 0 ° C, then diisopropylethylamine (0.287 ml, 1.65 mmol) is added dropwise. Diphenyl chlorophosphate (0.342 ml, 1.65 mmol) is added dropwise to the resulting zhepto-brittle solution, the reaction mixture is held at O for 30 million. Then diisopropylethylamine (0.313 ml, 1.80 mmol) is added and a solution of 3-methylthio-1- - / 3-mercaptoethyl / pyridinium chloride (0.398 g, 1.80 mmol) in 0.70 ml of dry DMF. Approximately 1 minute after the addition is complete, the precipitate is separated from the reaction mixture and then cooled to a temperature of -10 ° C, which is maintained for 10 minutes, resulting in a solid orange mass. This solid is triturated with acetonitrile and the residue is separated by filtration. The residue is washed with acetonitrile ,. then with acetone and dried under vacuum, whereby a product (0.455 g, 55%) is obtained as a cream-colored solid. The collected filtrate is evaporated, a yellow oil is obtained, which is transferred to a minimum volume of acetonitrile and cooled to 0 ° C, which is maintained for 30 minutes. By filtering this mixture, an additional 1.139 g of product is obtained in the form of a light yellow solid. The total yield of 0.594 g (72%).

C. / 5K, 68 / -3- 2- / 3-Methylthiopyridine / ethylthio-6- 1- / K / -oxyethyl-7-ox-1-azabicyclo 3.2.0 hept-2-en-2 - carboxylate.

 In the mixture of para-nitrobenzl / 5R, 6S / - -3-2- / 3-methylthiopyridinio / ethylthio - -6-1- / K / -oxyethyl / -7-oxo-1-azabicyclo f3.2.o hept-2 -en-2-carboxylate chloride (0.551 g, 1.0 mmol) and 10% palladium on charcoal (0.55 g) in 25 ml of phosphate buffer (0.05 M, pH 7.4) are added 5 ml THF

and 25 ml of ether. This mixture is subjected to hydrogenation (Parr) at a pressure of 2.812 kg / cm for 1 hour. The reaction mixture is then filtered through Celite and the filter cake is washed with water and ether. The aqueous phase is separated and washed with an additional amount of simple effect (3 x). After removing the residual organic solvents under vacuum, the aqueous solution is cooled to 0 ° C and the pH is maintained at 7.0 with a saturated aqueous solution of NaHCO3. This solution was immediately applied to a C-18 reverse phase chromatographic column. As a result of elution with water and subsequent lyophilization of the appropriate fractions, 0.25 g of a bright yellow solid is obtained. This material is again purified using high pressure reversed phase liquid chromatography, whereby the desired product (0.210 g, 55%) is obtained as a light yellow solid.

Example 10. Preparation of 3-C2- - / 1- / 2,6-dimethylpyridinium / ethylthio - -6 "i-ji- / K / -oxyethyl-7-oxo-1-azabicyclic / 3.2.0 / hept- 2-en-2-carboxylate

A. 1- / 2-Mercaptoethyl / -2,6-dimethyl pyridine methanesulfonate.

A mixture of 2,6-dimethylpyridine (19.2 ml of 0.165 mol) and methane (mono) sulfonic acid (3.27 ml, 0.050 mol) is stirred for 42 hours at 100 ° C. under nitrogen atmosphere. After cooling to 25 ° C, the reaction mixture was diluted with ether (45 mp) and water (30 ml). The two layers are separated and the organic layer is extracted with water (2 x 5 ml). The aqueous layers were combined, filtered through a Celite pad, rinsed with ether (2 x 15 ml), treated under vacuum to remove traces of organic solvents, and drained onto the top of the column (3.0 x 12 cm) from (C-bondapac C- 18. As a result of elution with a mixture of 3% acetonitrile and 97% water, 2.5 g of the crude title compound was lyophilized in the form of a syrup after it was lyophilized. It was again purified by high pressure liquid chromatography (| C-bondapac C-18); As a result, 0.90 g (7%) of the desired compound is obtained. of the connections.

Q j 0 5

Q

five

0

B.Paranitrob11Il-3- 2- / 1- / 2., 6-Diphenyl spiridium / ethyl o} -6c. - / R / -oxyethylJ-7-oxo-1-azabicyclo-3.2.0 / hept-2-e-11-carboxylate-di-1 IL-phosphate.

In a cold () solution of paran t-robenznl-6oS-1- / K / -oxyethyl-3-3, 7-d11-oxo-1-azabitsnklo / Z.2.O / heptan-2-cl-bauxilate (0.658 g, 1 , 89 mmol) in acetonitrile (6 mp), which is maintained under a nitrogen atmosphere, diisopropylethylamine (0.394 mp, 2.26 mmol) and diphenyl chlorophosphate (0.468 ml, 2.26 mmol) are added. The reaction mixture is stirred for 30 minutes and treated with a solution of 1- / 2-mercaptoethyl--2 / 2, b-dimethylpyridinium methane sulfonate (0.720 g, 2.73 mmol) in acetonitrile (3 ml), then diisopropyl ethyl amine (0.994 MP, 2.26 mmol). The reaction mixture was stirred for 2 hours, diluted with cold (С C) water (27 mp) and poured onto the top gr, column (2.5 x 9.0 cm) from (K-bondapac C-18. As a result of elution mixtures of acetonitrile-water and lyophilisate of the corresponding fractions give 0.92, g (65%) of the desired compound.

C.3- 2- (1- / 2,6-dimethylpyridine // ethylthio1 -6oi- G1- / I / -oxyethyl-7-I

-oxo-1-azabicyclo / З.2.0 / hept-2-e-2-carboxylate.

In solution.paranitrobenzyl-3- 2- - / 1- / 2,6-dimethylpnridine // ethylthis) - -6od- 1- / K / -oxyethyl-7-oxo-1 -azl bicyclo / 3.2.0 / hept-2-ene-2-carboxylate of diphenyl phosphate (0.80 g, 1.07 qmol) in a wet tetragran shrofuran (42 ml) was added ether (42 bp), potassium phosphate buffer of monobasic sodium hydroxide (0.15 M , pH 7.22, 21 ml) and 10% palladium on charcoal (0.8 g). The resulting mixture is subjected to hydrogenation for 1 hour at a pressure of: 2.812 kg / cm and a temperature of 23 ° C and filtered through a Celite pad. The two layers are separated and the organic layers are extracted with buffer (3x8 ml). The aqueous phase is combined, washed with ether (50 ml), treated under vacuum to remove traces of organic solvent, and poured onto the top of the column (3.0 x X 10.2 cm) from a 1-C-18 bonapzka. By elution of the column with a mixture of 5% acetonitrile — 95% water and freeze-drying of the corresponding fractions, the desired compound (0.2A6 g, 63%) was obtained as a yellowish powder.

Calculated: C 55.51, H 6.47; N 7.19.

 1,5H.jO

Found: C 55.1A; H 6.23; N 6.46.

Example 11. Getting / 5R, 6S / - -3- | 2- / 2-methyl-3-methyl-imidazo / -ethylthio-6- 1- / E / -oxyethyl-7-oxo-1-aabicyclo 3.2.oj hept-2-ene-2-carboxylate

A. 2-Methylthio-3-methyl-1/2-mercapto-ethyl / imidazolium trifluoromethanesulfonate.

Trifluoromethane (mono) sulfonic acid (1.38 MP, 0.015 mol) is added dropwise to 2-methylthio-1-methylimidazole (4.0 g, 0.03 mol) at 0 ° C under a nitrogen atmosphere. Ethyl sulfide (0.9 ml, 0.015 mol) is then added, the mixture is heated to 55 ° C under nitrogen and held under these conditions for 24 hours.

20 10% palladium on charcoal. This mixture is subjected to hydrogenation (Parr) at a pressure of 2.461 kg / cm for 55 minutes. The reaction mixture is then filtered over Celite, and the reaction mixture is triturated with ether (3x), and the residue is ne-25 The lather grate is washed with water and taken up in acetone, filtered and whith simple ether. The aqueous phase is separated,

ryvaet. The result is a product

cooled to OC, and the pH is maintained at 7.0 with a saturated aqueous NaHCO solution. After removing the residual organic solvents under vacuum, the aqueous solution is applied to a C-18 reverse phase chromatography column. After elution with water and then with a mixture of 8% acetonitripe, after lyophilization of the corresponding fractions, 0.25 g of solid is obtained. This material is again purified by high-pressure reverse-phase liquid chromatography, whereby the desired product (0.114 g, 19%) is obtained as an off-white solid. Example 12. Preparation of / 5R, 6S / (4.2 g, 82%) as a semi-crystalline solid, which is used as such without subsequent JQ purification.

B. Paranitrobenzyl / 5H, BZ / -3- 2- - / 2-methylthio-3-methylimidazolio / ethyl-thio-6- / 1- / K / -oxistil-7-oxo-1-azabicyclo 3.2.0J hept- 2-ene-2-carboxylate of diphenyl phosphate.

To the solution of paranitrobenzyl / 5Н, 65 / - -6-1- / К / -oxyethyl-3,7-dioxo-1-aza-bicyclo 3.2,0j heptane-2-carboxylate (1.40 g, 4.0 mmol) in 50 ml of dry acetonitrile at 0 ° C under nitrogen atmosphere, dropwise propyl ethylamine (0.76 ml, 4.4 mmol) is added dropwise and cooled to OC, and the pH is maintained at 7.0 using saturated aqueous NaHCO solution. After removing the residual organic solvents under vacuum, the aqueous solution is applied to a C-18 reverse phase chromatography column. After elution with water and then with a mixture of 8% acetonitripe, after lyophilization of the corresponding fractions, 0.25 g of solid is obtained. This material was again purified by high-pressure reverse-phase liquid chromatography, whereby the desired product (0.114 g, 19%) was obtained as an off-white solid. Example 12. Preparation / 5R, 6S / by that diphenylchlorophosphate (0.91 ml, ...,

4.1 mmol). After stirring the reaction, -3-2- (3-aminopyridino / ethylthio) -6 mixture at room temperature, 1- (H) -oxyethyl-7-oxo-1-azabi-ticure, for 1 h, diiso- Lo 3.2.03 Hept-2-en-2-carboxylate.

propylethylamine (0.76 mp, 4.4 mmol), and then a solution of 2-metntio-3-methyl-1- / 2-mercaptoethyl / imidazolium trifluoromethanesulfonate (2.0 g, 5.9 mmol) is added dropwise in 5 ml of acetonitrile. The reaction mixture is kept at room temperature for

A. 3-Amino-1- / 2-mercaptoethyl / pyridinium chloride.

3-Aminopyridine (1.50 g. About. 016 mol is placed in 15 ml of 1 methanolic solution of HC1 and the resulting solution is added to form hydrochloride as an oil. 3-aminopyridine (1.32 g , 0.015 mol) and ethylene sulfide (0.89 ml, 0.013 mol), and the resulting mixture is heated (oil per bath) to a temperature of 60-

1.5 hours and then concentrated in vacuo to give a gum. This resin was transferred to water and applied to a C-18 reverse phase column. As a result of being zoned with water.

then with a mixture of 20% acetonitrile — water, and finally with a mixture of 30% acetonitrile — water, after lyophilization of the corresponding fractions, the desired product (0.90 g, 30%) is obtained as a light yellow solid.

C. / 5K, 65 / -3- 2- / 2-methylthio-3-methylimidazolio / ethylthio (-6-1- / K / -oxyethyl) -7-oxo-1-azabicyclo (3.2.6 hept- -2-en-2-carboxylate.

Into a solution of paranitrobenzyl / 5R, 6S / - ./2-methylthio-3-meths1Imidazolio/-ethyltib-6- / 1- / K / -oxyethyl-7-oxo-1-azabicyclo 3.2.0J hept-2-en-2 -carboxylate diphenyl phosphate (1.20 g, 1.56 mmol) in a mixture of 70 mp THF, 70 yugh of ether and 31 ml of phosphate buffer (0.05 M, pH 7.4) was added 1.2 g

10% palladium on charcoal. This mixture is subjected to hydrogenation (Parr) at a pressure of 2.461 kg / cm for 55 minutes. The reaction mixture is then filtered over Celite, and the grille is washed with water and ether. The aqueous phase is separated,

cooled to OC, and the pH is maintained at 7.0 with a saturated aqueous NaHCO solution. After removing the residual organic solvents under vacuum, the aqueous solution is applied to a C-18 reverse phase chromatography column. After elution with water and then with a mixture of 8% acetonitripe, after lyophilization of the corresponding fractions, 0.25 g of solid is obtained. This material is again purified by high-pressure reverse-phase liquid chromatography, whereby the desired product (0.114 g, 19%) is obtained as an off-white solid. Example 12. Getting / 5R, 6S / ...,

-3-2- / 3-aminopyridine / ethylthio / -6- - 1- / H / -oxyethyl-7-oxo-1-azabicyclo 3.2.03 hept-2-ene-2-carboxylate.

A. 3-Amino-1- / 2-mercaptoethyl / pyridinium chloride.

3-Aminopyridine (1.50 g. O, 016 mol) is placed in 15 ml of 1 methanolic solution of HC1 and the resulting solution is added, resulting in a hydrochloride in the form of oil. Z-aminopyridine (1.32 g, 0.015 mol) and ethylene sulfide (0.89 ml, 0.013 mol) are added to this oil, and the resulting mixture is heated (oil on a bath) to a temperature of 60 ° C.

231A

65 ° C under nitrogen, which is maintained for 2 hours. Another equivalent of ethylene sulfide (0.89 ml, 0.015 mol) is added and heating is continued

o - t at 55-65 ° C for 65 hours. The reaction mixture is programmed and then transferred to water (25 ml). The aqueous solution is applied to the reversed-phasol: ro (-C-18 gato-graph, which is then eluted with water. Substitution of the appropriate fractions gives the desired product (1.26 g, 44%) as a colorless viscous oil.

B. Para-nitrobenzyl 5R, - / 3-aminopyridinio / ethyltib-6-1- (K) -oxyethyl-3-oxo-1-azabicyclo 3.2.0 hept-2-en-2-carboxylate

the mixture is hydrogenated (Parr) at a pressure of 2.812 kg / cm for 1 hour. The resulting mixture is filtered through a Celite pad, and the filter cake is washed with water and a simple EFF1F. The aqueous phase is separated, washed with ether (2), and then the residual volatile materials are removed under vacuum. The aqueous solution was immediately applied to the C-18 Span Phase Chromatography Column, which was eluted with water. As a result of the lyophilization of the corresponding fractions, 0.45 g of an off-white solid is obtained. This material is again purified using high pressure reverse phase liquid chromatography, in

nilphosphate.

Para-nitrobenzyl / 5R, 6S / -20 solution results in obtaining the desired pro-6- / 1- / I / -oxyethyl / -3,7-dio-cco-1-azaduct (0.123 g, 35%) as solid bicyclo / 3.2.0 / heptane-2-carboxylate ester of ivory.

(0.696 g, 2.0 mol) in 10 ml of dry Example 13. Production of (5R, 6S / acetonitrile at 0 ° C in an atmosphere of azo-3-1 / 5) -methyl-2- (1-pyrvidium) / et l- diisopropyl-25 thio-b- (1- (K) -oxyethyl 1-7-oxo-1-azabicyclo [3 .2.0] hept-2-en-2-carboxylate (A) "and / 5К, 65 / 3- 1- / К / -methyl-2- / 1- -pyridinium / ethylthioZ-6- 1- / R / -OKCH- ethylZ-7-oxo-1-azabicyclo / 3.2.0 / - hept-2-en-2-carboxylate (B).

A. d1-1- / 2-mercapto-2-methylethyl / pi30

ethylamine (0.382 nl, 2.2 mmol), then diphenyl chlorophosphate (0.457 ml, 2.2 mmol). After mixing at

A solution of 3-amino-1- / 2-mercaptoethyl / pyridinium chloride (0.475 g, 2.5 mmol) in

1 ml of dry DMF, then an additional amount of diisopropylethylamine (0.435 ml, 2.5 mmol). The reaction mixture was kept at 0 ° C for 1.5 hours, and then concentrated under varidium methanesulfonate, dl-l- / 2-Mep-capto-1-methylethyl / pyridinium methaxulphonate.

Methane (mono) sulfonic acid (1.95 ml, 0.030 mol) is slowly added to the cold pyridine (7.83 ml, 0.097 mol).

Methane (mono) sulfonic acid (1.95 ml, 0.030 mol) is slowly added to the cold pyridine (7.83 ml, 0.097 mol).

40

and the resulting mixture was stirred at 40 ° C for 15 minutes, treated with dl-propylene sulfide (2.59 ml, 0.033 mol) and stirred at 60 ° C under nitrogen atmosphere for 90 hours. Pyridine was removed under vacuum, the residue was mixed with water and cleaned up. The resulting resin was transferred to acetonitrile-water (1: 1) and applied to a C-18 reverse phase chromatography column. By elution with water, then with a mixture of 20% acetonitrile-water and subsequent lyophilization of the appropriate fractions, the desired product is obtained (0.730 g, 50%) as a solid using chromatography (beige-colored liquid. On high-pressure chromatography,

C. / 5H, 65 / -3- / 2- / 3-aminopyridone-bondudec C-18). The corresponding fracio / ethylthio / -6- / 1- / K / -oxyethyl / -7-oxy-1-azabicyclo / 3.2.0 / hept-2-en-2-carboxylate.

To the mixture of Para-nitrobenzyl / 5R, 6S / - -3- / 2- / 3-aminopyridinio / ethylthio / -6- - / 1- / R / -oxi-ethyl / -7-oxy-1-azabicyclo / 3.2.0 / hept-2-ene-2-carboxylate diphenyl phosphate (0.730 g, 1.0 mmol) and

55

50

10% palladium on charcoal (0.7 g) in 25 ml of phosphate buffer (0.05 M, pH 7.4) was added 8 ml of THF and 20 MP ether. Then this

These compounds are combined and lyophilized, resulting in 61-1- / 2-mercapto-2-methyl 1-ethyl / pyrhenium methanesulfonate (1.14 g, 15%) as a colorless syrup.

Calculated: C H 6.71 N 4.91; S 22.47.

C.) H, 5 Shz5g 2NgO

Found: C, 37.49; H, 6.85; N 4.86; S 22.09.

Dl-1- / 2-MepKanTo- -1-methylethyl / pyridine methanesulfonate is also obtained.

2f

0

the mixture is hydrogenated (Parr) at a pressure of 2.812 kg / cm for 1 hour. The resulting mixture is filtered through a Celite pad, and the filter cake is washed with water and a simple EFF1F. The aqueous phase is separated, washed with ether (2), and then the residual volatile materials are removed under vacuum. The aqueous solution was immediately applied to the C-18 Span Phase Chromatography Column, which was eluted with water. As a result of the lyophilization of the corresponding fractions, 0.45 g of an off-white solid is obtained. This material is again purified using high pressure reverse phase liquid chromatography, in

thirty

redinium methanesulfonate; dl-l- / 2-Mep-capto-1-methylethyl / pyridine methaxulphonate.

Methane (mono) sulfonic acid (1.95 ml, 0.030 mol) is slowly added to the cold pyridine (7.83 ml, 0.097 mol).

chromatography (high pressure liquid chromatography,

0

and the resulting mixture was stirred at 40 ° C for 15 minutes, treated with dl-propylene sulfide (2.59 ml, 0.033 mol) and stirred at 60 ° C under nitrogen atmosphere for 90 hours. Pyridine was removed under vacuum, the residue was mixed with water and purified by chromatography (high pressure liquid chromatography

Bonapac C-18). The corresponding fraction

five

0

These compounds are combined and lyophilized, resulting in 61-1- / 2-mercapto-2-methyl 1-ethyl / pyrhenium methanesulfonate (1.14 g, 15%) as a colorless syrup.

Calculated: C H 6.71 N 4.91; S 22.47.

C.) H, 5 Shz5g 2NgO

Found: C, 37.49; H, 6.85; N 4.86; S 22.09.

Dl-1- / 2-MepKanTo- -1-methylethyl / pyridine methanesulfonate is also obtained.

(0.82 g, 11%) as a colorless syrup.

Calculated: C 39.11; H 6.56; N 5.07.

SdH, 5 NO, Sj- 1,

Found: C 39.13; H 5.92; N 5.20.

B. / 5K, 65 / paranitrobenzyl - / K, 8 / -methyl-2- / 1-pyrindium / -ethyl2, 812 kg / cm and a temperature of 23 ° C. The two layers are separated and the organic layer is extracted with water (3x7 ml). The aqueous layers were combined, filtered through Cellit pad, washed with simple EFF1-FOM (3 X 20 ml) and subjected to chromatography on a C-18 bondapac in the form of a column (2.5 x 9.5 cm) using TH-J-6-1- / R / -oxyethy1G1-7-oxo-1-aza-10 by the formation of water as an eluting bicyclo [3.2.0] hept-en-2-carboxy-latephenyl phosphate.

In cold (О С) solution of / 5R, 6S / paranitrobenzyl-6- 1- / R / -3,7-dioxygenant, as a result of which 0.18 g (63%) of a mixture of diastereoisomers is obtained. The two diastereoisomers are separated by liquid chromatography of high-α-1-azabicyclo / 3.2.0 / heptane-2-carc j pressure (prep. Bondagac G-18) of boxylate (0.523 g, 1.5 mmol) in acetone using water as eluonitrile (6 ml) which contains a solvent: an isomer with a small

in nitrogen atmosphere, add diiso-retention time, 0.068 g (23%),

propipethylamine (0.314 ml, 1.8 mmol), compound B, an isomer with more extended then diphenyl chlorophosphate (0.373 mp, 20 retention time, 1.8 mmol). The reaction mixture was re-0.081 g (28%), compound A,

mix 30 minutes and process rast-PRI me R 1A, Getting / 5R,

by a thief d1-1 / 2-mercapto-2-methylethyl / pi-b5 / 3- 2- (5) - / 1-pyridium / -1- / 5 / - ridinium methanesulfonate (0.539 g, -cyclohexylthio-6- 1- / R / -oxy diethyl 2, t6 mmol) in acetonitrile (2 ml) and 25 7-oxo-1-azabicyclo [3.2.0] hept-2-diisopropylethylamine (0.314 mp, -en-2-carboxylate (A) and / 5R , 6S / 31, 8 mmol). The reaction mixture is pere-- 2- / R / - / 1-pyridinium (J-1- / R / -cyclome.) Taken at 0 ° C for 1 h, once the hexylthioz-6- 1- / R / -oxoxylp3- 7-okbavl with cold water (0 ° C, 25 ml) of co-1-azabicyclo / 3,2,0 / hept-2-en-2 and subjected to chromatography on the bond-0 carboxylate (B).

A. dl-1- / 2-Mecapto-1-cyclogekpeke C-18, having the form of a column (2.5 X 8.5 cm), using 25-50% acetonitrile in water as eluting solutions, as a result of which 1.07 g (97%) of the title compound as a yellowish powder after lyophilization.

Calculated: C 57.52; H 6.10; N 5.59; S 4.27,

C, jH "N, 0 ,, SP HfO

Found: C 57.76, H 4.96; N 5.36;

35

40

Se / Pyridine Methansulphonate,

Methane (mono) sulfonic acid (0.65 ml, 0.01 mol) is added dropwise to pyridine (2.42 ml, 0.03 mol) with cooling. The mixture was stirred under nitrogen for 10 minutes, treated with dl-cyclohexene sulfide (1.377 g, 85% purity, 0.0102 mol) and stirred for 25 hours. Excess pyridine was removed under vacuum, and traces were distilled off using water. . The residue is mixed with water and

Methane (mono) sulfonic acid (0.65 ml, 0.01 mol) is added dropwise to pyridine (2.42 ml, 0.03 mol) with cooling. The mixture was stirred under nitrogen for 10 minutes, treated with dl-cyclohexene sulfide (1.377 g, 85% purity, 0.0102 mol) and stirred for 25 hours. Excess pyridine was removed under vacuum, and traces were distilled off using water. . The residue is mixed with water and

S 4.35,

C. / 5R, 6S / 3- l- / R and S / methyl-2- / 1-pyridine / ethylthio} -6- 1- / K / -oxy- is subjected to chromatography through column ethyl / -7-oxo- 1-azabic clo (3,2,0) / hept-, well from prep. Bondapac C-18 (5 x 13 cm) -2-ene-2-carboxy at, using 6-2% acetonitrile

In solution (5Я, 78) paranitrobenzip in water as an eluting solution, 8 / methyl 2- (1-pyridium / ethyl), after lyophilization, thioZ-6-1 / K / -oxyethyl-7-oxo is obtained -1-aza-colorless syrup, 1.57 g (53%), bicyclic / 3.2.0 / hept-2-ene-2-carboxy-lateniphenyl phosphate (0.60 g, 0.82 mmol) in wet tetrahydrofuran (33 ml) ether (33 mp) is added, potassium phosphate buffer is monobasic

sodium hydroxide (17 ml, 0.15 N,

Calculated: C, 46.88; H 6.88j N.

C, iH ,,

Found: C 46.61-, H 6.46; N 4.65.

B. / 5R, 6S / parapanobenzyl-3-C2pH 7.22) and 10% palladium at 55 or 8 / - / 1-pyridium -1- / K or in spring angle (0.60 g). The resulting 8 / -cyclohexylthio-6- / 1- / I / -oxy-result is the mixture subjected to hydrogenation for 1 h under pressure

ethyl / -7-OXO-1-az abicyclo [3.2.0] hept-2-ene-2-carboxylate diphenyl phosphate,

2,812 kg / cm and a temperature of 23 ° C. The two layers are separated and the organic layer is extracted with water (3x7 ml). The aqueous layers were combined, filtered through a Cellite pad, washed with simple EF1-FOM (3 x 20 ml) and subjected to chromatography on a C-18 bondapac in column form (2.5 x 9.5 cm) using water as an eluting

35

40

Se / Pyridine Methansulphonate,

Methane (mono) sulfonic acid (0.65 ml, 0.01 mol) is added dropwise to pyridine (2.42 ml, 0.03 mol) with cooling. The mixture was stirred under nitrogen for 10 minutes, treated with dl-cyclohexene sulfide (1.377 g, 85% purity, 0.0102 mol) and stirred for 25 hours. Excess pyridine was removed under vacuum, and traces were distilled off using water. . The residue is mixed with water and

 in water as an eluting solvent, after lyophilization a colorless syrup is obtained, 1.57 g (53%),

Calculated: C, 46.88; H 6.88j N.

C, iH ,,

Found: C 46.61-, H 6.46; N 4.65.

B. / 5R, 6S / para-nano-benzyl-3-C2- / R or 8 / - / 1-pyridium -1- / K or 8 / -cyclohexylthio-6- / 1- / I / -oxyethyl / -7-OXO- 1-az abicyclo / 3.2.0 / hept--2-en-2-carboxylate diphenyl phosphate,

In cold (0 ° С) solution of / 5R, 6S / - paranitrobenzyl-6-1- / K / -oxyethyl-3,7-dioxo-1-azabicyclo / 3.2.0 / heptane-2-carboxylate (1 , 37 g, 3.93 mmol in acetonitrile (15 ml), which is maintained under a nitrogen atmosphere, diisopropylethylamine (0.822 ml, 4.7 mmol) and diphenyl chlorophosphate (0.979 MP, 4.7 mmol) are added. The resulting solution is stirred 30 min and treated with a solution of B1-1- / 2-mercapto-1-cyclohexyl / pyridine methanesulfonate (1.64 g, 5.66 mmol) in acetonitrile (4.7 yl), and then diisopropylethylamine (0.822 ml, 4, 7 mmol.) The reaction mixture was stirred at 0 ° C for 1 h, dispersed Cold (0 ° C) water (75 mp) and chromatographed on prep bonponac C-18 using 25-30% acetonitrile in water as an eluting solvent; 1.9 g (53%) is obtained after lyophilization of the corresponding fractions desired compound.

Calculated: C 59,16i H 5.35 j N 5.31.

C ,, H ,, o SP-HjO

Found: C 58.95; H 5.15, N 5.57.

C. / 5R, 6S / -3- 2- / R shsh 5 / - / 1-pyridinium / 3-1- / I or 5 / -cyclohexyl-6-f1- / K / -oxy-ethN-7 -oxo-1-aza bicyclo / 3.2.0 / hept-2-en-2-carboxylate.

In solution / 5К, 68 / para-nitrobenzyl- or 8 / - / 1-pyrcinium / -1- / I or 5 / -cyclohexylthio-6-1- / H / -oxy-ethyl-7-oxo-1-azabicyclo73.2.0 (1) hept-2-en-2-carboxylate diphenyl phosphate (1.85 g, 2.34 mmol) in wet tetrahydrofuran (96 ml), ether (96 mp), potassium phosphate buffer of monobasic sodium hydroxide (0, 15 M, pH 7.22, 50 ml) and 10% palladium on charcoal (1.9 g). The resulting mixture is subjected to hydrogenation at a temperature of 23 ° C, a pressure of 2.812 kg / cm for 1.25 hours. The organic layer is separated and washed with water (3 x 20 mp). The aqueous solutions are filtered through a Celite pad, washed with ether (2 X 60 ml), processed under vacuum to remove traces of organic solvents and subjected to chromatography on a column of prep. Bondapac C-18 (4.5 X 9 cm) using 0-5%

five

about

Q

0

acetonitrile in water as elui. solvent, after lyophilization get 0,705 g (76%) of a mixture of d asteroismers. The diastereoisomers are separated using high pressure liquid chromatography (prep. Bonapac C-18) using acetonitrile (4%) in water as the eluant of the solvent, diastereomer with a short retention time, compound A (0, 29 g, 31%).

Calculated: C 56.59; H 6.55, N 6.60; S 7.55.

 2N.O

Found: C, 56.83; H 6.47 N 6.59; S 7.43.

A diastereomer is also obtained with a longer retention time, compound B (0.35 g, 38%).

PRI me R 15. A. / 5K / Allyl-3- -G / 2-pyridinioethyl / thioZ- / 68 / - 1K / -oxy-ethyl-7-oxo-1-azabicyclo / 3.2.0 / - hept-2-ene-2-carboxylate diphenyl phosphate,

In solution / 5К / allyl-3,7-dioxo- / 65 / - / 1К / -oxoethyl -1-azabicyclo-p.2. OZheptan- / 2K / -carboxylate (473 mg, 1.87 mmol) in C1Csy (6 ml) at a temperature of about -10 ° C under a nitrogen atmosphere, diisopropininsTfmaMHH (0.42 ml, 2.4 mmol) is added, followed by diphenyl chlorophosphate (0.50 ml, 2.4 mmol). The mixture was stirred at 0 ° C for 30 minutes, then cooled to -15 ° C. An oily suspension of H- / 2-mercaptoethyl / pyridinium chloride (527 mg, 3.00 mmol) in CH, CN (1 ml) containing 5 drops of DMF, then di-isopropylethylamine (0.42 ml, 2.4 mmol; 1). The mixture is stirred for 30 minutes and then diluted with water (20 ml). This mixture is directly cleaned in reverse -phase chromatographic silica gel column, elution was carried out with water (200 mp), mixtures 10% (100 mp), 20% CH CN / HjO (100 ml), 30% CH 3 CH / H O (100 ml), 40% (100 ml). Compliant co btfayut fractions, the organic solvent was removed in vacuo and lyophilized to give 786 mg (1.26 mmol, yield 67.3%) of the title compound as a brownish powder.

B. / 5R / -3- / 2-pyrlinioethyl / thio - - / 68 / - / 1H / -oxyethyl-3-oxo-1-aza-bicyclo 3.2.0 hept-2-en-2-carboxylate.

In solution / 5К / allyl-3- / 2-pyridinioethyl / thio3- / 63 / - 71К / -oxyethyl-7- -oxo-1-azabicyclo p. 2.b hept-2-Å-2-carboxylate phenylphosphate (156 mg 0.25 mmol) in (2 ml) are sequentially added at a temperature of

Q

approximately 22 ° C. solution of 2-ethylhexanoate potassium in EtOAc (0.5 M, 0.6 ml, 0.3 mmol), triphenylphosphine (15 mg, 0.057 mmol) and tetrakistriphenylphosphine palladium (15 mg,

0.013 mmol). The mixture is peremegpat at a temperature of about 22 ° C under nitrogen atmosphere for 2 hours. After adding anhydrous (7 ml) by filtration, the precipitate is separated, washed with anhydrous (7 ml) and dried under vacuum, resulting in 101 mg of a basalt solid. It is subjected to purification using a reversed-gas chromatographic column (C-18 PrepPAC, 12 g, Waters Associates) using water as eluent. The appropriate fractions (fractions 7 through 12, each with a volume of 20 kilograms) are collected and lyophilized, resulting in 53 mg (0.16 mmol, 64% yield) of the desired compound in a yellowish powder.

Example 16 Preparation of - / K-Methyl Thiomorpholine / Ethylthio - -6oL- / l - / E / oxyethyl-7-oxo-1-aza-bicyclo 3.2. 0 hept-2-en-2-carboxylate.

A, N-methyl-M- / 2-mercaptoethyl / thiomorpholine methanesulfonate.

To the pre-cooled (ice per bath) N-methylthiomorpholine (5.00 g, 42.7 mmol), methane (mono) sulfonic acid (1.47 mp, 20.5 mmol) and ethylene sulfide (1.30%) are added. (1.30 MP, 21.4 MP, 21.4 mmol.) The mixture is heated to a temperature of 65 ° C, which is maintained for 24 hours, and diluted with water (25 ml). The aqueous solution is washed with diethyl ether (Ex X 25 ml) is treated under vacuum and poured onto a silica gel reverse-phase chromatography column, the title compound is eluted with water. The appropriate fractions are collected and evaporated, resulting in the formation of a thiol e oil (4.80 g, yield 86%)

B.Paranitrobenzyl-3- 2- / K-methyl-β-thiomorpholinium diphenyl phosphate / ethyl-. - / K / -oxyethyl-7-oxo-1-azabicyclo 3 .2.03 hept-2-en-2-carboxylate.

Cold (ice bath) solution of paranitrobenzyl-god. - fl - / K / -oxyethyl} - -3, 7-dioxo-1-azabicyclo 3.2, VOC heptane-2-carboxylate (557 mg, 1.60 mmol in CH-jCN (8 ml) is treated dropwise with diisopropylethylamine (0.336 MP, 1.92 mmol) and diphenyl chlorophosphate (0.400 ml, 1.92 mmol) and stirred for 30 minutes. The reaction mixture is treated with M-methyl-N- / 2-mercaptoethyl / thiomorphinium methanesulfonate (893 mg, 2.29 mmol) in (4 mp) and diisopropylethylamine (0.336 mp, 1.92 mmol) and stirred for 30 min. The solution was diluted with water (20 ml) and poured onto a column reverse phase chromatography silica gel. The desired compound is eluted with a mixture of 50% acetonitrile — water. The appropriate fractions are combined, treated under vacuum for 2 hours and lyophilized to give the desired compound (1.01 g, yield),

C. 3-C2- / N-methyl-thiomorpholine / - erlUlTio-6 - / - / K / -oxyethyl-7-oxy-1-azabicyclo 3.2.0 hept-2-en-2-carboxylate.

Para-nitrobenzyl-3- {2- / N- -methylthiomorpholinium diphenylphosphate / - ethylthioT-6ot- / l- / K / -oxyethylZ-7-oxo-1-azabicyclo 3.2.0 hept-2-en-2- -carboxylate solution (1.31 g, 1.7 mmol) in 0.1 M phosphate buffer pH 7.4 (48.8 mp), tetrahydrofuran (20 ml) and diethyl ether (20 ml) is subjected to hydrogenation over 10% Pd / C (1.5 g) on a Parr vibrator for 1 hour at a pressure of 2.812 kg / cm. The reaction mixture is diluted with diethyl ether (40 mp) and the phases are separated. The organic phase is washed with water (2 X 5 ml). The aqueous phases are combined, filtered through filter paper, washed with diethyl ether (2 x 20 ml) and treated with vacuum. An aqueous solution of slime on the column by reversed phase chromatography on silica gel and the desired carbapenem is eluted with a mixture of 5% acetonitrile — water. The appropriate fractions are co-cooled and lyophilized, resulting in the desired compound being obtained in the form of an amorphous solid (203 mg, 31%),

31U2473732

Example 17. Getting / 5R, 6S / - 20% aitonitripe - water and a mixture of JU) t

acetonitrile - water and subsequent liofilizatsin appropriate fractions

3- 2- / 1-methylmorpholio-ethylthio-6- - / K / -1-hydroxyethyl-7-oxo-1-azab11cyclo 3.2.6 hept-2-ene-2-carboxypate.

A. 1-mbT1SH-1- / 2-mercaptoethyl / morpholinium trifluoromethanesulfonate.

In N-methylmorpholine (3.29 ml, 0.030 mol), trifluoromethane (mono) sulfonic acid (1.337 ml, io co-1-azabicyclo 3.2.o3 hept-2-en-car- 0.015 mol) is added dropwise at 10 ° C, then sulfide bokoshat.

 Ethylene (0.89 ml, 0.015 mol). Semi-In solution of paranitrobenzyl / 5R, 6S / resulting from the yellow-brown-3- 2- / 1-methylmorpholino / ethylthio-6 solution is heated (oil per bath) - / K / -1-hydroxyethyl-7-oxo-1- azabits} 1kdo 50-60 ° C under nitrogen and seeded with 3.2.0Jhept-2-en-2-carboxpatapitate under these conditions for 18 hours. Fly-phenyl phosphate (0.360 g, 0.49 mmol)

g get the desired product (0.360 g, 40%) as an amorphous solid.

C. / 5K, B8 (-3-2-1-methylmorpholino) 6- / "/ - 1-oxneethyl-7-occhy materials are then removed under vacuum, and the resulting residual oil is transferred to 10 ml of water. Water

The solution is washed with diethyl 20 tetrahydrofuran and 20 ml with ether (3x5 ml) and then with ethyl ether. This reaction organic solvent is removed - the mixture is hydrogenated under vacuum. Obtained as a result of (Parr) at a pressure of 2.250 kg / cm in

The aqueous solution was applied to a C-18 reverse phase chromatography column, which was then eluted with water, a mixture of 5% acetonitrile — water, and a mixture of 10 Z acetonitrile — water. By evaporation of the appropriate fractions, a more solid substance is obtained, which is dried under vacuum (PjOj), and the desired product is obtained (1.92 g, 41%).

B. Para-nitrobensh1 / 5K, 68 / -3-G2- / 1-methylmorpholino / ethylthio-1-hydroxyethyl-7-oxo-1-azabicyclo-3.2.0 hept-2-en-2-carboxylate diphenyl phosphate,

Into the solution of paranitrobenzyl / 5R, 6S / - -6-1K / -1-hydroxyeth-1 -3,7-dioxo-1-aza-bicyclo 3,2.d heptane-2-carboxylate (0.348 g, 1.0 mmol) in 25 ml of dry acetonitrile is added dropwise di-isopropylethylamine (0.191 ml, 1, t mmol) and then diphenyl chlorophosphate (0.228 ml, 1.1 mmol) at 0 with under nitrogen atmosphere. After stirring at 0 ° C for 1 h, diisoproethyl 1ML (0.226 ml, 1.3 mmol) was added, the resulting enol phosphate and then 1-methyl-1 - / 2-mercaptoethyl / morphopinium trifluoromethanesulfonate (0.373 g, 1.2 mmol The reaction mixture is stirred at

1.5 hours at room temperature and then concentrated under vacuum. The residual material is transferred to water and then carried onto a C-18 reverse phase column. As a result of elution with water, then with a mixture of

co-1-azabicyclo 3.2.o3 hept-2-ene-carboococate.

obtain the desired product (0.360 g, 40%) as an amorphous solid.

C. / 5K, B8 (-3- 2- (1-methylmorpholino) etatio 6- (") -1-oxnethyl-7-okv 13 MP phosphate buffer (0.05 M, pH 7.4) was added 0, 36 g of 101 st palladium on charcoal, 20 ml

for 1 h. The mixture is filtered through Celite, and then the filter pad is washed with water and diethyl ether. The aqueous phase is separated; its pH is maintained at 7.0 by the addition of phosphate buffer with a pH of 7.4. After removing the residual organic solvents under vacuum, the aqueous solution is applied to a C-18 reverse phase chromatography column. As a result of water elimination and lyophilization of the corresponding fractions, 0.130 g of an amorphous solid is obtained. This material is subjected to repeated purification with slop and reverse phase high pressure liquid chromatography, resulting in a pure product (0.058 g, 34%) as an amorphous solid.

Example 18. Preparation of / 5R, 6S / -, 4-dimethyl piperazine / ethyl acetate-6-1 / R / oxy x-7-oxo-1-aza-bicyclo 3.2.0 hept-2-carboxylate ,

A, 1- / 2-Acetylthioethyl / -1,4-dimethylpiperazinium bromide.

A solution of 2-bromoethyl ethyl acetate (2.20 g, 0.012 mol) and 1, A-dimethyl piperazine (1.95 ml, 0.014 mol) in acetone (A ml) is transferred at 50 ° C for 65 hours. After cooling to a liquid phase decanted, leaving a gum that is triturated twice with diethyl ether, resulting in a hydroscopic

personal yellowing powder (3.2 g, 90%).

B. 1,4-Dimethyl-1- / 2- -mercaptoethyl / piperazinium bromide hydrochloride.

A solution of 1- / 2-acetylthioethyl / -1,4-dimethyl piperazinium bromide (1.1 g, 3.7 mmol) in 6 N hydrochloric acid solution (4 Mji) is heated to 80 ° C under azo atmosphere and under these conditions incubated for 1 h. The solution is concentrated under reduced pressure, resulting in a white powder (0.41 g, 38%).

Calculated: C 31.03 J H 7.16; N 9.05; S 10.35.

CjH oNjSBrCl HjO

Found: C, 31.62; H 7.46; N 9.19, S 10.19.

C./5K.65/Paranitrobenzyl/3- {2- - / 1,4-dimethyl-1-piperazinium / -ethyl-Li / K / hydroxyethyl-7-OXO-1-aza bicyclo 3.2.0 hept-2- en-2-carboxy-latephenyl phosphate.

In cold (0 ° С) solution / 5R, 6S / - paranitrobenzyl-6-G1 / K / hydroxyethyl 1- -3,7-dioxo-1-azabicycloGz.2.03hep-, tan-2-carboxylate (0.465 g,

1.33 mmol) in acetonitrile (2 ml), maintained under a nitrogen atmosphere, diisopropylethylamine (0.33 ml 1.59 mmol) is added. The reaction mixture is stirred for 30 minutes and treated with a suspension of 1,4-dimethyl-1- - / 2-mercapto-ethyl / -pipresinium bromide hydrochloride (0.40 g, 1.37 mmol) in acetonitrile (3 ml) - water. (1 ml) and diisopropyl-ethylamine (0.278 ml, 1.59 mmol). After stirring for 18 hours, cold water (15 ml) was added to the mixture. The resultant solution is subjected to chromatography over Prep-Pak-500 / C-18 (Waters-Associate #} tc) in the form of a column (2.5x7.5 cm) using 25-35% acetonitrile in water as eluting solvents , as a result, after freeze-drying, a yellowish powder is obtained.

(0.50 g, 50%).

D. / 5R, 6S / -3- 2- / 1,4-dimethyl-1-pyperoxynium / ethylthio-6-1- / K / -oxy-ethyl-7-oxo-1-azabicyclo 3.2. 0 - hept-2-en-2-carboxylate,

In solution / 5К, 65 / paranitrobenzyl-3- 2-11,4-dimethyl-1-piperazinium - ethylthio-6-1- (K) -oxyethyl-7-ok-c0-1-azabicyclo 3.2.0 hept-2-ene-2- carboxypath diphenyl phosphate (0, A7 g, 0.623 mmol) in wet tetrahydro5

R

0

five

furan (25 ml) was added with di-, ztirate ether (25 ml), potassium phosphate buffer monobasic sodium hydroxide (13 ml, pH 7.22) and 10% palladium on charcoal (0.47 g) . The resulting mixture is hydrogenated at 23 ° C and a pressure of 2.812 kg / cm for 1 hour. The two layers are separated and the organic layer is extracted with water (2 x 7 ml). The aqueous layers were combined, filtered through a Celite pad, washed with diethyl ether (2 X 15 ml) and subjected to chromatography on a column (2.5 x 9.5 cm) using water as an eluting solvent, resulting in a 0.097 yield. g (43%) of the desired product after lyophilization.

Example 19. Preparation of / 5R, 6S / -, 4,4-trimethyl-1-piperazine / -ethylthio-6-} R-hydroxyethyl-7-ox-1-azabicyclo / T.2.0 / hept-2 -en-2- -carboxylate.

A. 1- / 2-Acetylthiostil / -1,4,4-trimethylpiperazinium bromide iodide.

A suspension of 1- / 2-acetylthioethyl / -1, 4-dimethyl piperazinium bromide (1.48 g, 5.0 mmol) in isopropyl alcohol (10 ml) is treated with methyl iodide (0.373 MP, 6.0 mmol) and heated to a temperature of 55-60 C, which is maintained for 30 hours. The solvents are evaporated under reduced pressure, the residue is triturated in hexane, and the solid is separated by filtration (1.85 g). The solid is dissolved in hot water (8 mp) and the solution is diluted with auer tone to turbidity (70-80 ml). As a result of two consecutive crystallizations, 1.5 g of the title compound (68%) are obtained with a mp. 220-225 ° C (decomposition).

Calculated: C, 30.08; H, 5.51; N 6.38.

From „1

Found: C, 30.48; H, 5.53; N 6.86.

B. 1- / 2-Mercaptoethyl / -1,4, A-tri-met- shppiperazinibischloride.

A mixture of 1- / 2-acetylthiostil / -1,4,4-trimethyl piperazinium bromide iodide (1.84 g, 4.19) and 6 N hydrochloric acid solution (15 mp) is heated to under nitrogen and held under these conditions 2, 5 h. The solution is concentrated under reduced pressure until dry. The solid is suspended in water.

they take another 2h. A portion of the acetonitrile was removed under reduced pressure, and the resulting red mixture was subjected to chromatography using 25-75% acetonitrile in water as the elution solvent, to obtain a yellowish powder (1.4 g) after lyophilization. The powder is dissolved in water and the solution is passed through a column (1.2 x 58 cm) of permutite S -1 C1 using water as an eluting agent. Lyophilization of the appropriate fractions gives 1.17 g of powder, which is again purified using a Preprap-500 / C-18 column, a yellowish powder (0.80 g, 53X) is obtained by freeze-drying the corresponding fractions.

HE

H

Hjc

S-A-Rp

where R, is hydrogen, paranitrobenzyl or allyl protective group if there is also a counterion

R, - pyridium or substituted pyridine, imidazolium, thiomorpholynium, piperazinium, morpholines)

A - ethylene, unsubstituted. Or substituted by methyl, cyclohexyl,

outcome, from ketoester general formula II

37U2473738

HE. HS-A-Rj

I Ngde A and Rj have the indicated values

.hh interaction of ketoester total form HJC 1 G I p-. pt of simula II with diphenylchlorophosphate in the presence of diisopropylethylamine in vUUn in medium

where Rj is the indicated inert organic solvent group

diphenylchlorophosphate, diieopropylstil- and a mercaptan derivative of the rest a fina, inert organic grow in one stage.

the voritel and Mercaptan derivative, Q 2, Method pop. 1, distinguished by being removed if necessary and by the fact that the process is carried out

carboxy-protecting group, about t l and h a - at a temperature of from -15 C to room n and with the fact that, for the purpose of simplified

The puppy of the process, as a pro- duct, 3, The method according to claim 2, the distinguishing mercaptan uses a thiol common and that the process is carried out

Formula III at a temperature of -15 to + 15 ° C.

Claims (3)

Claim 1. The method of obtaining derivatives of carbalenem of the general formula I, he n H ₃ C N— ^C0 ° R ₁ where R, is hydrogen, a paranitrobenzyl or allyl protective group in the presence of also a counterion; - pyridinium or substituted pyridinium, imidazolium, thiomorpholinium, piperazinium, morpholinium; A is ethylene, unsubstituted. Or substituted with methyl, cyclohexyl, starting from a keto ester of the general formula II where Rj is the indicated protecting group, diphenyl chlorophosphate, diisopropylethylamine, an inert organic solvent and a Mercaptan derivative, with the removal of a carboxy protecting group, if necessary, I owe the fact that, in order to simplify the process, a thiol of general formula III is used as a mercaptan derivative HS-A-Rj, where A and R ₂ have the indicated meanings, and the reaction of the keto ester of general formula II with diphenylchlorophosphate in the presence of diisopropylethylamine in an inert organic solvent and a mercaptan derivative is carried out in one step. 2. The method according to claim 1, characterized in that the process is carried out at a temperature of from -15 ° C to room temperature. 3. The method according to claim 2, characterized in that the process is carried out at a temperature of from -15 to +15 C.