FI92487C - Process for the preparation of 2- (heterocyclylthio) carbapenem derivatives useful as drugs - Google Patents

Description

y ^ HrjV

Process for the preparation of 2- (heterocyclylthio) -carbapenem derivatives useful as medicaments

For the preparation of 2- (heterocyclylthio) carbapenem derivatives according to the invention 5

The invention relates to a process for the preparation of compounds which can be used as medicaments, which can be represented by the formula (I): 10 HO H CH3

\ / I

C C-H

/ \ / \ 15 H3c CH_CH C-S-R «I I II (I)

C_N_C

/ \ O COOR5 20 [j ossa

Ra represents a group of formula 1-8 or 1-9: CH2-CH2-CHN-R3

C

30 I (1-8) o 35 o CH2— (1-9)

I I

- CH N - R 3 / ch 2 »292487 and wherein: R 3 is a hydrogen atom or a C 1 -C 4 alkyl group; 5 R5 is a hydrogen atom or a carboxy-protecting group which is selected from the group-måstå which kåsittåå following: alkoxycarbonyloxyalkyl groups, sykloalkyylialkoksikarbonyylioksialkyyliryhmåt, sykloalkyylioksikarbonyylioksialkyyliryhmåt, 10 cycloalkylcarbonyloxyalkyl groups, which are optionally substituted alkyl-substituted yhdellå lower alkyyliryhmållå, aliphatic acyloxy-methyl groups, ylemmåt aliphatic acyloxyalkyl groups, terpene penyloxycarbonyloxyalkyl groups and 5-alkyl-substituted (2-oxo-1,3-dioxolen-4-yl) alkyl groups; 15 and for the preparation of pharmaceutically acceptable salts and esters of these compounds.

Penicillins form a well-known class of antibiotics, 20 members of which have been widely used in human and veterinary medicine for many years. Chemically, penicillins have a common β-lactam structure, commonly referred to as a "penam", which can be represented by the following formula:

S

/ \ • _ · · III (A) • -N_ · 30 /

Although penicillins still play an important role in pharmacy, 35 new pathogenic, often resistant to penicillins

II

92487 3 The development of capable bacterial strains has made the search for new types of antibiotics increasingly necessary.

In recent years, considerable interest has been given to compounds having a carbapenem structure, i.e., compounds having a carbon atom at the 1-position at the sulfur atom and having a double bond between the 2- and 3-position carbon atoms in the original panam structure. This carbapenem structure can be represented by the following formula: 10 / \ * 5.

15 I I II (B) 7 * -N- ·! // 4

O

20 These penam and carbapenem structures form the basis for the designation of semisynthetic penicillin derivatives in accordance with the recommendations of the International Union of Pure and Applied Chemistry (IUPAC). These designations are generally accepted by experts in the art and are also used herein. The numbering system used herein is in accordance with formula (B) above.

The best known of the known carbapenem derivatives is the compound termed "thienamycin", the semisynthetic name of which is 2- (2-aminoethylthio) -6- (1-hydroxyethyl) carbapen-2-em-3-carboxylic acid. Although thienamycin is known to have remarkably potent and extensive bactericidal activity, its chemical stability in the human body is poor, which limits its applications. Thus, numerous attempts have been made in the art to modify the chemical structure of tie-namycin in order to improve its stability while maintaining or improving its excellent activity. However, there is still a continuing need in the art for the development of new carbapenem antibiotics with improved properties.

The present invention provides a new class of carbapenem-derivative-5 women which have excellent absorption and metabolic stability (as seen in larger amounts recovered from urine) and which are additionally broad-spectrum for killing bacteria and have very low toxicity. The invention makes it possible to make pharmaceutical preparations containing said derivatives which can be administered to humans and animals.

Of the prior art publications known to the applicants, the following are believed to be the closest to the present invention: U.S. Patent Nos. 4,640,799 and 4,665,170 describe carbapenem compounds having the group -S-A-N + in the 2-position. "A" may denote various groups and N + denotes a quaternized nitrogen-containing heterocyclic group attached to the group A via its quaternary nitrogen atom. These compounds differ from the compounds of the present invention in that if the compounds of the present invention have a quaternized nitrogen-containing heterocyclic group having a quaternary nitrogen atom, then this group is not attached to the remainder of the molecule. for the rest of the week. Via 25 carbon atoms belonging to the rocyclic group.

European Patent Publication 126,587 describes a series of carbapenem compounds having a pyrrolidinylthio group in the 2-position. These compounds differ from the quaternary nitrogen-containing compounds of the invention in that the compounds already known in the art are not quaternary nitrogen-containing compounds of the quaternary nitrogen and differ from those of the present invention which do not. nature. The activity of certain compounds described in these prior art publications may be

Si Π 'Π * 7 (I Η Ο / 5 is good, but they are believed to be less well absorbed in vivo compared to the compounds of the present invention.

Another state of the art is discussed below.

The carbapenem derivatives disclosed in EP-170073 have heteroarylalkylthio substituents at the 2-position of the carbapenem core and each heteroaryl group contains a quaternary nitrogen atom. The present compounds have heterocyclylthio groups without a quaternary nitrogen atom at this position, and thus these inventions differ.

The carbapenem derivatives disclosed in EP-242134 have heteroarylalkylthio substituents at position 15 of the carbapenem nucleus and each heteroaryl group contains a quaternary nitrogen atom substituted with an amino or alkylamino group. The present compounds have heterocyclylthio groups without a quaternary nitrogen atom at this position, and thus these inventions differ.

The carbapenem derivatives disclosed in EP-184844 have heteroarylalkylthio substituents at the 2-position of the carbapenem core and each heteroaryl group contains a quaternary nitrogen atom. The present compounds have heterocyclylthio groups without a quaternary nitrogen atom at this position, and thus these inventions differ.

The carbapenem derivatives disclosed in EP-186057 have heteroarylalkylthio substituents at the 2-position of the carbapenem core and each heteroaryl group contains a quaternary nitrogen atom. The present compounds have heterocyclylthio groups without a quaternary nitrogen atom in this position, and thus the present inventions differ.

The carbapenem derivatives disclosed in EP-184843 have 35 heteroarylalkylthio substituents at the 2-position of the carbapenem nucleus and each heteroaryl group contains a quaternary nitrogen atom.

6 c A o 7 y l. Lt u /

The present compounds have heterocyclylthio groups without a quaternary nitrogen atom at this position, and thus these inventions differ.

Some carbapenem derivatives of EP-126587 have a pyrrolidinylthio substituent at the 2-position of the carbapenem core and each pyrrolidinyl group has a carbamoyl or alkylcarbamoyl group at the 5-position of the pyrrolidine core. The present compounds have a 5-pyrrolidon-3- or 4-ylthio group in this position. The present invention 10 differs from this reference.

Some carbapenem derivatives of EP-182213 have a pyrrolidinylthio substituent at the 2-position of the carbapenem core and each pyrrolidinyl group has a carbamoyl or alkylcarbamoylalkyl group at the 5-position of the pyrrolidine core. The present compounds have a 5-pyrrolidon-3- or 4-ylthio group in this position. The present kek-sintd differs from this reference.

Some carbapenem derivatives of claim 7 of EP-160876 have a 5-pyrrolidon-2-ylthio substituent at the 2-position of the carbapenem core and a chlorine or fluorine substituent at the 1-position of the carbapenem core. Some carbapenem derivatives, e.g., those set forth in Example 85 herein, have a 5-pyrrolidone- (2,3 or 4) -alkylthio substituent at the 2-position of the carbapenem core and a chloro-substituent at the 1-position of the carbapenem core. The present compounds have a 5-pyrrolidon-3 or 4-ylthio group in the 2-position of the carbapenem core and a methyl group in the 1-position of the carbapenem core. The present compounds differ from this reference.

Some of the carbapenem derivatives disclosed in EP-167139 (list of compounds) have heteroarylthio substituents at the 2-position of the carbapenem nucleus and each heteroaryl group contains a quaternary nitrogen atom having an alkyl group in the carbamene base and no substituent at the carbamene base. The present compounds have 35 heterocyclylthio groups without a quaternary nitrogen atom carbapenic

II

92487 7 in the 2-position of the nucleus and the methyl group in the 1-position and differ from this reference.

Some of the carbapenene-5 derivatives described in GB-2128187 (FI-78094) (Examples 2-25) have heteroarylalkylthio substituents at the 2-position of the carbapenem nucleus, and each heteroaryl group contains a quaternary nitrogen group having an alkyl group having an alkyl group. The present compounds have a heterocyclylthio group without a quaternary nitrogen atom at the 2-10 position of the carbapenem nucleus and differ from the compounds of this reference.

Some of the carbapenem derivatives disclosed in EP-72710 (FI-76339) (Examples 2,7,8,9,11,12,16,17,18 and 20) have a pyrrolidin-3-ylthio substituent at the 2-position of the carbapenem core but not 15 substituents at the 1-position of the carbapenem core. The present compounds have a 5-pyrrolidon-3- or 4-ylthio group in the 2-position of the carbapenem nucleus and a methyl group in the 1-position and differ from the compounds of this reference.

Some of the carbapenem derivatives disclosed in EP-160391 (FI-83419) have a pyrrolidin-3-ylthio substituent at the 2-position of the carbapenem core and various substituents at the 1-position of the carbapenem core. The carbapenem derivative (1) in Example 8 has a pyrrolidin-5-ylthio substituent at the 2-position of the carbapenem core and a methyl group. 25 in its 1-position. We believe that the present compounds having a 5-pyrrolidon-3- or 4-ylthio group in the 2-position of the carbapenem nucleus and a methyl group in the 1-position differ from the compound (1).

Structurally, the closest compound to the compounds of the present invention from the above and following technical documents is the compound of the FI-83419 patent (1). Although the compounds of the present invention are closely related to compound (1), representative present compounds, for example, the compounds of this application in Examples 36, 39, 40 and 41, have a stronger antibacterial activity against a variety of 35 clinically important microorganisms than the compounds of FI Patent 1 83419. ).

8 92487

Some of the carbapenem derivatives disclosed in EP-102239 (FI-80887) (Example 9c, 10c, 12b, 13d, 16e and 19b) have a pyrrolidin-3-ylthio substituent at the 2-position of the carbapenem core but no 5 substituents at the 1-position of the carbapenem core. The present compounds have a 5-pyrrolidon-3- or 4-ylthio group in the 2-position of the carbapenem nucleus and a methyl group in the 1-position and differ from the compounds of this reference.

Some of the carbapenem derivatives disclosed in EP-186525 (FI-81576) (Examples 1-8) have a pyrrolidin-3-ylthio group in the 2-position of the carbapenem core and have substituents at positions 1 and 5 of the pyrrolidine nucleus but no substituents at the 1-position of the carbapenem core. The present compounds having a 5-pyrrolidone-3- or 4-15 ylthio group in the 2-position and methyl in the 1-position of the carbapenem core differ from the compounds of this reference.

Some of the carbapenem derivatives disclosed in EP-165384 (FI-84826) (Examples 2,7,8,9,11,12,16,17,18 and 20) have a pyrrol-20-din-3-ylthio substituent at the 2-position of the carbapenem core and not substituents at the 1-position of the carbapenem core. The present compounds having a 5-pyrrolidon-3- or 4-ylthio group in the 2-position of the carbapenem nucleus and a methyl group in the 1-position differ from the compounds of this reference.

. 25

Some of the carbapenem derivatives disclosed in EP-202048 (FI-81577) (Examples 1,4,5,10,11,12 and 14) have a pyrrolidin-1-yl-1-iminoethylthio substituent at the 2-position of the carbapenem core and the pyrrolidine ring has a substituent 3- position. The present compounds have a 5-pyrrolidon-3- or 4-ylthio group in the 2-position of the carbapenem nucleus and differ from the compounds of this reference.

Some of the carbapenem derivatives disclosed in EP-235823 (FI-843446 and 861721) (Examples 2 and 3) have a heterocyclyl-35-alkylthio substituent having a quaternized nitrogen atom at the 2-position of the carbapenem core. The present compounds having a 5-yl 9,92487 pyrrolidone-3- or 4-ylthio group at the 2-position of the carbapenem nucleus differ from the compounds of this reference.

Some carbapenene-5 derivatives disclosed in EP-280771 (FI-875135) (examples 3,4,7,8,12,13,14,20,21,22,23,24,25,31, 32,33,34 , 35,37,39,42,43 and 45) is a pyrrolidin-3-ylthio group having a substituent at position 5 or at the 2-position of the carbapenem nucleus of the pyrrolidide core. The present compounds having a 5-pyrrolidon-3- or 4-ylthio group in the 2-position of the carbapenem nucleus and having no substituent in the 5-position of the pyrrolidine nucleus differ from the compounds of this reference.

The invention provides a pharmaceutical composition comprising an effective amount of an antibiotic in admixture with a pharmaceutically acceptable carrier or diluent, said antibiotic being at least one of the compounds of formula (I) and pharmaceutically acceptable compounds of these compounds.

Furthermore, the invention provides a method of treating or preventing a microbial infection in a mammal, which may be a human, by administering to it an effective amount of an antibiotic which is at least one of the compounds of formula (I) and pharmaceutically acceptable derivatives thereof. 25 ta selected compound.

In the compounds of the present invention in which R3 represents an alkyl group, this alkyl group may be a straight-chain or branched alkyl group having 1 to 4 carbon atoms, and may be exemplified by methyl, ethyl, propyl, isopropyl, butyl, isobutyl. butyl, t-butyl, of which methyl, ethyl, propyl and butyl are preferred, with methyl being generally most preferred.

R5 represents a hydrogen atom or a protecting group which can be removed under chemically reasonable conditions, for example by means of a chemical reducing reagent or by catalytic reduction, or _____ - - · T—.

f 'nt r r- ΓΊ y ά H 0! 10 which can be removed by a biological reaction, e.g. in vivo, to produce a carboxy group. There is no restriction on the nature of this protecting group, as long as the resulting compound is used for medical purposes, the compound is pharmaceutically acceptable, meaning that the activity of the compound is not reduced (or that the activity is not reduced). or that its toxicity has not been increased (or that its toxicity has not been increased in an inappropriate manner) compared to the corresponding compound of formula (I) in which R5 is a hydrogen atom. However, if the compound is intended to be used in non-medical applications, for example as an intermediate in the preparation of other compounds, then this restriction does not apply and the protecting group can be selected solely on the basis of process-related criteria.

The compounds of the present invention may contain a basic group and thus may form acid addition salts. The nature of these salts and acids is not critical to the invention, provided that when the compound is used for medical purposes, the salt must be pharmaceutically acceptable, which is well known in the art to mean that it does not have less (or significantly less) activity or not greater (or significantly greater) than the free base activity or toxicity. However, if the compound is intended to be used in other applications, for example as an intermediate in the preparation of other compounds, then this restriction does not apply.

Examples of acids which may form such salts or which can provide a balancing anion include inorganic acids such as hydrochloric acid, hydrobromic acid, hydroiodic acid, phosphoric acid, sulfuric acid or nitric acid; organic sulfonic acids such as methanesulfonic acid, ethanesulfonic acid, benzenesulfonic acid or p-toluenesulfonic acid; as well as organic carboxylic acids such as oxalic acid, tartaric acid, citric acid, maleic acid, succinic acid, acetic acid, benzoic acid, mandelic acid, ascorbic acid, lactic acid, and gluconic acid.

Examples of the cations present in the compounds of the present invention represented by the symbol M include metal atoms, especially alkali metal atoms such as sodium and potassium atoms, and alkaline earth metal atoms such as the calcium atom; ammonium group; as well as cations derived from a trialkylamine such as triethylamine or another organic base such as procaine, dibenzylamine or Fenet-10-ylamine.

The compounds of formula (I) may exist as different isomers due to the presence of asymmetric carbon atoms. The present invention encompasses both the individual isomers and mixtures of two or more isomers. Such mixtures may be prepared by synthetic reactions or by stirring. If the individual isomer is the desired product, it may be prepared by a stereospecific synthetic method or may be prepared by separation of a mixture of isomers using separation techniques well known in the art. Particularly preferred are those compounds in which the 1-hydroxyethyl group at the 6-penem position is in the same configuration as thienamycin, i.e. it is 1 (R) -hydroxyethyl.

The (IR, 5S, 6S) configuration is preferred.

The following compounds of the invention are most preferred: 25 «

Pivaloyloxymethyl 2- (2-oxo-4-pyrrolidinylthio) -6- (1-hydroxyethyl) -1-methyl-1-carbapen-2-em-3-carboxylate, especially its pivaloyloxymethyl (1R, 5S, 6S) -2 - (2-oxo-4-pyrrolidinylthio) -6 - [(1R) -1-hydroxyethyl] -1-methyl-1-carbapen-2-eme-3-carboxylate-30 isomer; % (1-methylcyclohexan-1-yl) carbonyloxymethyl 2- (2-oxo-4-pyrrolidinylthio) -6- (1-hydroxyethyl) -1-methyl-1-carbapen-2-em-3-carboxylate, in particular its (1-methylcyclohexan-1-yl) carbonyl-oxyloxymethyl (1R, 5S, 6S) -2- (2-oxo-4-pyrrolidinylthio) -6 - [(IR) -1-12f; r ', / n «-» 7 ά 40 (hydroxyethyl] -1-methyl-1-carbapen-2-em-3-carboxylate isomer; 1- (Cyclopentyloxycarbonyloxy) ethyl 2- (2-oxo-4-pyrrolidinyl-5-thio) -6- (1-hydroxyethyl) -1-methyl-1-carbapen-2-em-3-carboxylate, in particular its 1- (cyclopentyloxycarbonyloxy) ethyl (1R, 5S, 6S) -2- (2-oxo-4-pyrrolidinylthio) -6 - [(1R) -1-hydroxyethyl] -1-methyl-1-carbapen-2-emetic 3-carboxylate isomer; As well as pharmaceutically acceptable salts of these compounds.

The process according to the invention is mainly characterized in that (a) HO H CH3 of formula (IV):

\ / I

C C-H

H3C CH CH C-R27 I I II (IV)

C_M_C

20 / \ ..

O COOR28 [wherein R1 is as defined above, R27 is a group of the formula -OR29, wherein R29 is a diphenylphosphoryl group; and R28 represents a 4-nitrobenzyl group]; The compound of formula 30 is reacted with: «a compound of formula (Vb): HS- (Ra) (Vb) 35 92487 13 [wherein Ra is as defined above, wherein only the active groups are protected, if necessary] to give a compound of formula 5 (VI): HO H CH 3

\ / I

C C-H

/ \ / \! 0 H3C CH_CH C-S-Rb I I II (VI)

C_N_C

O COOR2 8 [wherein R1, R28 are as defined above and Rb is said group of formula 1-8 or 1-9, wherein optionally any active group may be protected], 20 (b) after which, if necessary, the protecting groups are removed and / or the resulting compound is esterified and / or salified to give a compound of formula (I) or a pharmaceutically acceptable salt or ester thereof. 1

In more detail, these compounds can be prepared as described in Methods A and B below.

14 O O / · P 1-1 y Ih o /

Method A

This method can be illustrated by the following reaction scheme A: CO / C η y L · η ο / REAmOfCAAVIO A: 15 OH Η1

Step A 1 C "3 I 75 J - * - k 28 ° Hl 0 COOR28 (VII) OH R1 k \ / 0R23 CH3 'III Step A; - __» - L [+ (Va), (Yb |] "\ 90 cal 0 COOR28 (VIII) OH R1 ^ A ^ / S- (CH2 ^ -Rb CH3 j Step A3: o coor28 (VI) OH R1 / J ^ / s- (cH2irRa

CHo I I I

11 koOR5 II) 16 ο η. · ^ ^ 7 i-Η O /

In the above schemes, R1, R5, Ra, Rb, R28, R29 and 1 are as defined above.

In the compound of formula (Va), X "represents a balancing anion which is preferably a halogen atom (e.g. chlorine, bromine or iodine) or an alkylsulfonyloxy, arylsulfonyloxy or halosulfonyloxy group (e.g. methanesulfonyloxy, p-toluenesulfonyloxyethyloxy, trifluorosulfonyloxyethane, trifluoro ).

In step A1 of this reaction scheme, the starting compound of formula (VII) is reacted in the presence of a base with an active ingredient of an alkanesulfonic acid, arylsulfonic acid, dialkylphosphoric acid-dihydroxyphenic acid-sulfonic acid or diarylphosphoric acid or sulfonic acid, e.g. In step A2, the resulting compound of formula (VIII) is reacted without isolation with a mercaptan derivative of formula (Va) or (Vb) in the presence of a base to give a compound of formula (VI). The desired compound of formula (I) may then be prepared by removing the protecting group R28 from the carboxyl group of the compound of formula (VI), if necessary.

Examples of reactive derivatives of the reagent of formula R290H useful in step A1 include: alkane sulfonic anhydrides such as methanesulfonic or ethanesulfonic anhydride; arylsulfonic anhydrides such as benzenesulfonic or ε-toluenesulfonic anhydride; dialkylphosphoryl halides such as dimethylphosphoryl or diethylphosphoryl chloride; diarylphosphoryl halides such as diphenylphosphoryl chloride or diphenylphosphoryl libromide. Of these reagents, p-toluenesulfonic acid is preferred. polyanhydride or diphenylphosphoryl chloride. There is no particular restriction on the nature of the solvent to be employed, provided that it has no adverse effect on the reaction or on the reagents involved. Examples of suitable solvents include halogenated hydrocarbons, especially halogenated aliphatic hydrocarbons such as methylene chloride, 1,2-dichloroethane or chloroform; nitriles such as li ς * Γ *. / r * f * 114 · :, / 17 acetonitrile; amides such as Ν, Ν-dimethylformamide or Ν, Ν-dimethylacetamide. There are also no particular restrictions on the nature of the base to be used, as long as it does not adversely affect other parts of the Mole moiety, in particular the β-lactam ring. Preferred 5 bases that can be used in this reaction are organic bases such as triethylamine, diisopropylethylaraine or 4-dimethylaminopyridine.

The reaction can take place over a wide range of temperatures, and the precise reaction temperature is not critical to the invention. It is generally considered convenient to carry out the reaction at a relatively low temperature to avoid side reactions, usually at -20 to + 40 ° C. Likewise, the time required for the reaction may vary widely depending on many factors, notably the reaction temperature and the nature of the reagents. However, provided that the reaction is effected under the preferred conditions outlined above, a period of from 10 minutes to 5 hours will usually suffice.

The resulting compound of formula (VIII) does not need to be isolated prior to the next step in the reaction scheme. Thus, in step A2, the reaction mixture can be treated with a mercaptan derivative of formula (Va) or (Vb) in the presence of a base. The nature of the base used in the reaction is not critical, but preferred bases include organic bases such as triethylamine or diisopropylamine as well as inorganic bases such as potassium carbonate or sodium carbonate.

The reaction can take place over a wide range of temperatures, and the precise reaction temperature is not critical to the invention. However, it is generally preferred to carry out the reaction at a relatively low temperature, for example from -20 ° C to room temperature. Likewise, the time required for the reaction may vary widely depending on many factors, notably the reaction temperature and the nature of the reagents. However, if the reaction is carried out under the preferred conditions described above, a period of from 30 minutes to 5 days will usually suffice.

18 ο Γ \ r · ~ · r-η S iL 4 O /

After the reaction is complete, the desired compound of formula (VI) can be recovered from the reaction mixture in a conventional manner. For example, one suitable way to recover the product is to simply remove the solvent by distillation from the reaction mixture. The resulting compound can be further purified, if necessary, in a conventional manner, for example, by recrystallization, reprecipitation, or by a variety of chromatographic techniques, such as column chromatography or preparative thin layer chromatography.

10

Prior to or after such further purification, the carboxy protecting group may be removed if desired. This is preferably carried out without isolating the compound of formula (VI).

The final reaction step of Method A comprises the removal of the carboxy protecting group R28 from a compound of formula (VI) to give the corresponding carboxylic acid of formula (I), and, if desired, the conversion of the resulting free acid to another salt or ester. This step is optional, and it is clear that removal of the carboxy-protecting group is not always necessary or desirable, for example, if the compound of formula (VI) is a pharmaceutically acceptable ester within the scope of the present invention. If it is desirable to remove the carboxy protecting group, this can be done by conventional methods, the choice of which depends on the nature of the protecting group used.

If the protecting group can be removed by reduction, for example, if it is a haloalkyl group, an aralkyl group or a benzhydryl group, it can be removed by contacting it with a reducing agent. Haloalkyl groups such as 2,2-dibromoethyl. or in the case of 2,2,2-trichloroethyl, the preferred reducing agent is a combination of zinc and acetic acid. If the protecting group is an aralkyl group (such as a benzyl or p-nitrobenzyl group) or a benzhydryl group, it is preferably removed either catalytically from the reducing agent using hydrogen and a suitable catalyst such as platinum or carbon-bonded palladium; or by reducing with alkali metal sulfide li ♦ 0 0 A Η, η s ZHu 1 19 such as sodium sulfide or potassium sulfide. Whatever the reduction technique, the reduction is preferably carried out in the presence of a solvent, the nature of which is not critical as long as it does not adversely affect the reaction. Suitable solvents are, for example, alcohols (such as methanol or ethanol), ethers (such as tetrahydrofuran or dioxane), aliphatic carboxylic acids (such as acetic acid) or a mixture of one or more such organic solvents with water. The reaction temperature is not critical, but is normally from 0 eC to room temperature. The time required for the reaction will vary depending on the nature of the starting materials and reducing agents as well as the reaction temperature, but a period of from 5 minutes to 12 hours will normally suffice.

After the reaction is complete, the desired compound containing the free carboxy group can be recovered from the reaction mixture by conventional methods. In suitable techniques for recovering the product, for example: any insoluble material is separated off, after which the solvent is removed by distillation to give the desired product. If necessary, the product can be further purified by conventional methods, for example by recrystallization or by various chromatographic techniques, such as preparative thin layer chromatography or column chromatography.

The carboxy-25 group in the compound prepared as described above can, if desired, be converted to an ester group which hydrolyzes under physical conditions. This can be done by traditional methods. If R5 represents an ester which hydrolyses under physiological conditions, for example pivaloyloxymethyl, acetoxymethyl, phthalidyl, indanyl, methoxymethyl or 2-oxo-5-methyl-1,3-dioxolen-4-ylmethyl, then the formula (I) ) can be hydrolyzed in vivo under physiological conditions. Thus, such a compound can be administered directly to a patient without removing the protecting group.

35 20 G O / - 7 - L · ‘t O /

Method B

This method can be illustrated by the following Reaction Scheme B: Reaction Scheme B: 10 OH R '.

A A J-M

C Η] I | | Step B] J-N-L [+ (Yal (Yb |]

^ A / 3_ (CH2 ^ -Ra CHj I I I

J-N-k ς 0 COOR5 II) 21 O r '·' · 'r ~ 7> ΔΗ <,.> J

In the above formulas, R1, Ra, Rb, R2S, R30 and 1 are as defined above.

The compounds of the formula (IX) 5 used as starting materials in this reaction scheme can be prepared in Japanese Patent Application Kokai No. Sho 62-30781 as described.

In Step B1, a compound of formula (VI) may be prepared by reacting a compound of formula (IX) with a mercaptan compound of formula (Va) 10 or (Vb) in the presence of a base in an inert solvent. There is no particular restriction on the nature of the solvent to be employed, provided that it has no adverse effect on the reaction or on the reagents involved. Examples of suitable solvents include ethers such as tetrahydrofuran; Nitriles such as acetonitrile; fatty acid amides such as dimethylformamide; sulfoxides such as dimethyl sulfoxide; water; or a mixture of any two or more such solvents. The base used in the reaction is also not critical, as long as it does not affect other parts of the molecule, in particular the β-lactam ring. Examples of suitable bases include organic bases such as diisopropylethylamine, triethylamine, N-methylpiperidine or 4- (N, N-dimethylamino) pyridine; as well as inorganic bases, especially alkali metal carbonates such as potassium carbonate or sodium bicarbonate. The reaction can take place over a wide range of temperatures, and the precise temperature is not critical to the invention, although it is preferred to carry out the reaction at a relatively low temperature to prevent side reactions. In general, the reaction is conveniently carried out at a temperature in the range of -20 to + 40 ° C. The time required for the reaction may also vary widely, depending on many factors, notably the reaction temperature and the nature of the reagents. However, if the reaction is carried out under the preferred conditions described above, a period of from 5 minutes to 5 days will usually suffice.

After the reaction is complete, the desired compound of formula (VI) can be recovered from the reaction mixture by conventional methods. Likewise, compounds of formula (I) may be prepared, if desired, by deprotection of a compound of formula (VI) using the procedure described in Method A. The resulting compound can be further isolated and purified as described in Method A 5.

Compounds of formula (Va) and (Vb) which are also used as starting materials in the above reaction schemes can also be obtained by conventional and well-known methods for the preparation of this type of compound. In the case of compounds of formula (Vb) which are not otherwise available, they may be prepared as follows:

First, a compound of formula (X): L-Rc (X) [where L represents a leaving group such as a hydroxy group, a halogen atom (such as chlorine, bromine or iodine) or a sulfonyloxy group (e.g. methanesulfonyloxy, propanesulfonyloxy, trifluoromethyl) -thanesulfonyloxy or toluenesulfonyloxy) and Rc represents 2-oxopyrrol-dinyl as defined in connection with the group of formula (III)] to convert to the corresponding protected thio compound of formula (XI):

Rd-Rc (XI) 30 [where Rd represents a thio group protecting group such as an alkanoyl group (e.g. acetyl or propionyl group) or an aralkyl group (e.g. 4-methoxybenzyl, 3,4-dimethoxybenzyl, benzhydryl, triphenylmethyl or di). (4-methoxyphenyl) metyyliryhmåå)].

35 If L represents a hydroxy group, it may have been obtained

The Mitsunobu reaction, which can be carried out under conditions known per se,

II

92487 23 in the presence of diethyl azodicarboxylate, triphenylphosphine and thioacetic acid.

If L represents a halogenator or a sulfonyloxy group, the reaction can be carried out by reacting a compound of formula (X) with a sodium or potassium salt of Rd-SH.

The second step is a conventional hydrolysis reaction and can be carried out under conventional conditions known per se. For example, if Rd is an alkanoyl group, it can be removed under alkaline or acidic conditions with, for example, sodium hydroxide in aqueous methanol. If Rd is an aralkyl group, it can be removed, for example, with trifluoromethanesulfonic acid in the presence of trifluoroacetic acid and anisole.

15

The mercaptan compound of formula (Va) can be prepared as described in the following Reaction Scheme C: Reaction Scheme C: R2 -1- (CH7) m 71 Step C1 R31-S- (CH2) # - -r --- 11 R3-L; KH0

(CH 2) n NH

(XII) R 2 y-1- (CH 2) m 71 Step C 2 R 1 -SHCH 2 -I-R 1 -x-

(CH2) n-H

X 3

(XIIII R

R2 Y-1- (CH2 R31-S- (CH2] t-- ^ -1L.

(¾ — V3 _ '/ (XIV) R * 1 R2: V I (^ 2) m HS-S- (CH2) £ - <CH2ln-®N ~ R3 1 L X®

(Val RL

II

92487 25

In the above formulas, R 31 represents a mercapto protecting group such as an aralkyl group [exemplified by 4-methoxybenzyl, triphenylmethyl, benzhydryl, 3,4-dimethoxybenzyl or di (4-methoxyphenyl) methyl], an alkanoyl group (e.g. acetyl, propyl or propionyl); ) or an aromatic acyl group (e.g. an o- or 2-toluoyl group or a benzoyl group); L represents a leaving group such as a halogen atom (e.g. chlorine, fluorine or iodine) or a sulfonyloxy group (e.g. methanesulfonyloxy, toluenesulfonyloxy, trifluoromethanesulfonyloxy or fluorosulfonyloxy); and R 2, R 3, R 1, m, n and X * are as defined above.

The compound of the formula (XII) used as a starting material in this reaction scheme can be prepared by known methods, an example of which is described in Japanese Patent Application Kokai No.

15 Sho 60-233076.

In Step C1, a compound of formula (XIII) is prepared by reacting a compound of formula (XII) with a compound of formula R 3 -L (XV) in the presence of a base. Alternatively, when R 3 is a methyl group, the compound of formula (XII) is reacted with formaldehyde (usually as formalin or paraformaldehyde) and then sodium cyanoborohydride; or it can be reacted with formaldehyde and then the reaction product is catalytically hydrated with palladium on carbon; or it can be. May react with formaldehyde and formic acid by heating the reaction mixture.

The reaction is preferably carried out in the presence of an inert solvent. There is no particular restriction on the nature of the solvent to be employed, provided that it has no adverse effect on the reaction or on it. participating reagents. Examples of suitable solvents include amides, especially fatty acid amides such as Ν, Ν-dimethylformamide or Ν, Ν-dimethylacetamide; halogenated hydrocarbons, especially halogenated aliphatic hydrocarbons such as methylene chloride, 1,2-dichloroethane or chloroform; ethers such as tetrahydrofuran, dioxane or diethyl ether; nitriles such as acetonitrile; and a mixture of any one or more of such solvents with water.

The reaction can take place over a wide range of temperatures, and the precise reaction temperature is not critical to the invention. It is generally considered convenient to carry out the reaction at a temperature in the range of -20 to + 100 ° C, but the reaction is preferably carried out at a relatively low temperature to avoid side reactions. The time required for the reaction may also vary widely, depending on many factors, notably the reaction temperature and the nature of the reagents. However, if the reaction is carried out under the preferred conditions described above, a period of from 10 minutes to 2 days will usually suffice.

In step 2 of this reaction scheme, a compound of formula (XIII) is reacted with a compound of formula R * -X (XVI). This reaction is preferably carried out in the presence of an inert solvent. There is no particular restriction on the nature of the solvent to be employed, provided that it has no adverse effect on the reaction or on the reagents involved. Examples of suitable solvents include aromatic hydrocarbons such as benzene, toluene or xylene; halogenated hydrocarbons, especially halogenated aliphatic hydrocarbons such as methylene chloride, 1,2-dichloroethane or chloroform; amides, especially fatty acid amides such as Ν, Ν-dimethylformamide or • Ν, Ν-dimethylacetamide; ethers such as tetrahydrofuran, dioxane or diethyl ether; nitriles such as acetonitrile; and a mixture of one or more such solvents with water.

The reaction can take place over a wide range of temperatures, and the precise reaction temperature is not critical to the invention. General purpose. it is considered appropriate to carry out the reaction in the range of -20 to + 180 ° C

but the reaction is preferably carried out at a relatively low temperature to avoid side reactions. The time required for the reaction may also vary widely, depending on many factors, notably the reaction temperature and the nature of the reagents. However, if the reaction is carried out under the preferred conditions described above, a period of from 10 minutes to 3 days will usually suffice.

In step C3 of this reaction scheme, the compound of formula (XIV) prepared as described in step C2 is deprotected to give the compound of formula (Va).

The method for removing the protecting group varies depending on the nature of the protecting group R31. Thus, when R 31 is an aralkyl group, removal of the protecting group may be conveniently carried out by reacting a compound of formula (XIV) with trifluoromethanesulfonic acid in the presence of trifluoroacetic acid and anisole.

On the other hand, if R 31 is an alkanoyl group or an aromatic acyl group, the protecting group may be conveniently removed by treating the compound of formula (XIV) with hydrogen chloride or hydrogen bromide in a suitable solvent such as an alcohol (e.g. methanol or ethanol), water or water. e.g. in aqueous tetrahydrofuran or aqueous diethyl ether).

20

The reaction can take place over a wide range of temperatures, and the precise reaction temperature is not critical to the invention. It is generally considered expedient to carry out the reaction at a temperature in the range of -20 to + 200 ° C, but the reaction is preferably carried out at a relative temperature. 25 at low temperatures to avoid side reactions. The time required for the reaction may also vary widely, depending on many factors, notably the reaction temperature and the nature of the reagents. However, if the reaction is carried out under the preferred conditions described above, a period of 10 to 30 minutes to 24 hours will usually suffice.

After the reaction is complete, the desired compound of formula (Va) can be recovered from the reaction mixture in a conventional manner, for example, by removing the solvent from the reaction mixture by simple distillation. If necessary, the desired compound can be further purified by a conventional method such as reprecipitation ------- - J-92487 28 or by various chromatographic methods such as column chromatography or preparative thin layer chromatography.

In the compound of formula (Va), the counterion X "varies depending on the nature of the reagent used in step C3.

The compounds of the present invention have excellent antibacterial activity and a wide spectrum of activity. Likewise, they are able to resist β-lactamase. As determined by the agar plate dilution method, these compounds have been shown to be active against a wide variety of pathogenic microorganisms, including both Gram-positive bacteria (such as Staphylococcus aureus and Bacillus subtlis) and Gram-negia coli (Escherichia coli). flexneri.

15 Klebsiella pneuminiae. Proteus vulgaris. Serratia-lai it. e.g.

Serratia marcescens. Enterobacter species. e.g., Enterobacter cloacae. Salmonella enteritidis and Pseudomonas aeruginosa '), and thus can be used to treat diseases caused by such microorganisms in humans and other animals. Dehydropeptidase I inactivates thienamycin and its analogs in vivo in mammals, while the compounds of the present invention are much more stable in the presence of this enzyme and in their case the amounts recovered from the urine are high, so they have good biological activity. Likewise, their toxicity is low when tested in laboratory animals.

: 25

Table 10 shows the activity of a compound of the invention at the lowest inhibitory concentration (Mg / ml) against various bacteria.

30 φ 92487 29

Table 10

Compound Microorganism

from example AB C

5 _ 4 0.01 0.01 * 25

A: Staphylococcus aureus 209 10 B: Escherichia coli NIHJ

C: Pseudomonas aeruginosa 1001 0.01 *: not more than 0.01.

The above results indicate that the compounds of the present invention can be used to treat or control a variety of diseases caused by pathogenic bacteria.

The esters produced as described in Examples 7.8 and 9 were incubated at 37 ° C for 1 hour with horse serum, at which time MC values (minimum inhibitory concentration) were determined. These values were exactly the same as those obtained in Example 4 and shown above. This means that these esters are easily cleaved by the esterase after oral administration in the small intestine, are well absorbed through the gastrointestinal tract and achieve complete free acid activity.

The following are comparative data to demonstrate the effect of the compounds of the invention.

> 30 92487

Microbial Comp. A United. B Comb. C Comb. D Vert.yhd.

a <0.01 <0.01 <0.01 <0.01 <0.01 b <0.01 <0.01 <0.01 <0.01 <0.01 5 C <0.01 <0 .01 <0.01 <0.01 0.02 d <0.01 <0.01 <0.01 <0.01 0.05 e 0.02 <0.01 <0.01 <0.01 0 , 4 f 0.1 0.1 0.2 0.05 0.4 10

In the tables above, the abbreviations mean the following:

Comp. A: Compound of Example 4:

Sodium (IR, 5S, 6S) -2- (2-oxo-4-pyrrolidinylthio) -6 - [(IR) -15 1-hydroxyethyl] -1-carbapen-2-em-3-carboxylate

Comp. B: Compound of Example 7:

Pivaloyloxymethyl- (1R, 5S, 6S) -2- (2-oxo-4-pyrrolidinylthio) -6 - [(1R) -1-hydroxyethyl] -1-methyl-1-carbapen-2-20 emem-3- carboxylate

Comp. C: Compound of Example 8: (1-Methylcyclohexan-1-yl) carbonyloxymethyl- (1R, 5S, 6S) -2- (2-oxo-4-pyrrolidinylthio) -6 - [(IR) -1-hyd-; 25 roxyethyl 1-methyl-1-carbapen-2-em-3-carboxylate

Comp. D: Compound of Example 9: 1- (Cyclohexyloxycarbonyloxy) ethyl- (IR, 5S, 6S) -2- (2-oxo-4-pyrrolidinylthio) -6 - [(IR) -1-hydroxyethyl] -1-me-30 style 1-carbapen-2-eme-3-carboxylate

Reference compound: Reference compound of Example 8 of FI patent 83419 Sugimura: (1R, 5R, 6S) -6- [1 (R) -hydroxyethyl] -1-methyl-2 - [(3S) -pyrrolidin-3- ylthio] -carbapen-2-eme-3-carboxylic acid 35

II

92487 31 a: Staphylococcus aureus 209P JC-1 b: Escherichia colt NIHJ JC-2 c: Klebsiella pneumoniae 806 d: Serratia marcescens IAM1184 5 e: Proteus vulgaris 1420 f: Moreanella morganii 1510

The antibacterial effect was stripped with agar solution on nutrient agar at 107 cfu / ml for 8 hours after planting at a temperature of 37 ° C. The antibacterial effect was measured, as measured by MICSO, i.e. the minimum inhibitory concentration required to inhibit half of the microbial growth.

As for the ester compounds of Examples 7, 8 and 9 of this application, 15 of them were treated before being tested in this test with horse serum to release free carboxylic acid and the incubation time was 1 hour at 37 ° C. From these results, it can be seen that the present compounds had an unexpectedly strong antibacterial activity compared to the structurally closest prior art compound FI-20 publication 83419 (1), as shown in the following table.

The compounds of the present invention may be administered either orally or parenterally in the treatment of pathogenic; 25 Diseases caused by microorganisms in humans and other animals. The compound can be converted into any of the conventional forms of use. Suitable oral dosage forms include, for example, tablets, granules, capsules, powders, and syrups, while parenteral dosage forms include, for example, intramuscular or, more preferably, intravenous injectable solutions. set.

The compounds of the present invention are preferably administered parenterally, in particular by intravenous injection.

35 32 92487

The dose of a compound of the invention will vary depending on the age, body weight and condition of the patient, as well as the route of administration and the frequency of administration. However, the daily dose for an adult is usually 100-3000 mg of the compound and may be administered in a single dose or in divided doses.

5

Example 1

Sodium (1S, 5S) -2- (2-oxo-3-pyrrolidinyl) -6-β-β) -1-hydroxy-ethyl-1-carbapen-2-em-3-carboxylate 10,126 μΐ diisopropylethylamine and 150 μΐ of diphenylphosphoryl chloride was added dropwise to an ice-cooled solution containing 250 mg of 4-nitrobenzyl- (IR, 5R, 6S) -6 - [(IR) -1-hydroxyethyl] -1-methyl-2-oxo-1- carbapenam-3-carboxylate in 5 ml of dry gun-15 tonitrile, and then this mixture was stirred at 0-5 ° C for 1 hour. At the end of this period, 288 μΐ of diisopropylethylamine and a solution of 222 mg of 3-mercapto-2-pyrrolidinone (prepared by a method similar to that described in "Preparation 19") are added to the ice-cooled reaction mixture (prepared as described in "Preparation 19") and trifluoro) in acetonitrile.

The whole mixture was then allowed to stand in the refrigerator for 3 days. At the end of this period, the mixture was diluted with ethyl acetate, washed successively with aqueous sodium bicarbonate solution and aqueous sodium chloride solution, and then dried over anhydrous magnesium sulfate; 25 la. The solvent was then distilled off under reduced pressure to give a crude product which was dissolved in a mixture of 20 ml of tetrahydrofuran and 20 ml of 0.1 M phosphate buffer (pH 7.0) and hydrogenated at room temperature for 2 hours in the presence of 331 ml. mg 10% w / w palladium on carbon. At the end of this period, insoluble material was removed by filtration using Celite as a filtration aid and the filtrate was washed with diethyl ether.

The aqueous layer was concentrated by evaporation under reduced pressure and the residue was subjected to column chromatography using a Diaion HP-20AG column eluted with water. The eluate was concentrated by evaporation under reduced pressure and lyophilized to give the crude product.

This crude product was further purified using chromatography

II

92487 33

Lobar column (Merck Co., LiChroprep RP-8, size B) eluted with 3% v / v aqueous methanol. The eluate was concentrated by evaporation under reduced pressure and lyophilized to give 35 mg of the title compound as a colorless powder.

5

Ultraviolet absorption spectrum (H 2 O) λmax nm: 302.

Nuclear Magnetic Resonance Spectrum (D 2 O, 270 MHz) δ ppm: δ 1.00 (3H, doublet, J = 7.33 Hz); 1.11 (3H, doublet, J ** 6.60 Hz); 1.86-1.98 (1H, multiplet); 2.40-2.55 (1H, multiplet); 3.18 - 3.39 (4H, multiplet); 3.71 (1H, doublet of doublets, J = 9.16 & 6.60 Hz); 4.02 - 4.12 (2H, multiplet).

Example 2 Sodium (1R, 5S, δS1) -2- (2-oxo-4-pyrrolidinyl) 6 - [(1R) -1-hydroxyethyl] -1-methylcarbapen-2-em-3-carboxylate 126 μΐ of diisopropylethylamine and 150 μΐ of diphenylphosphoryl chloride were added dropwise to an ice-cooled solution containing 250 mg of 4-nitrobenzyl- (IR, 5R, 6S) -6 - [(IR) -1-hydroxyethyl] -1-methyl-2 -oxo-1-carbapenam-3-carboxylate in 5 ml of dry acetonitrile, and this mixture was stirred at 0-5 ° C for 1 hour. At the end of this period, 144 μl of diisopropylethylamine and 97 mg of 4-mer-30-capto * 2-pyrrolidinone (prepared as described in "Preparation 20") were added to the ice-cooled reaction mixture. The whole mixture was then stirred for 7 hours at 0-5 ° C and then allowed to stand overnight in the refrigerator. The reaction mixture was then diluted with ethyl acetate, washed twice with aqueous sodium chloride solution and then dried over anhydrous magnesium sulfate. The solvent was then removed by distillation under reduced pressure to give a crude product which was dissolved in a mixture of 34,92487 containing 20 ml of tetrahydrofuran and 20 ml of 0.1 M phosphate buffer (pH 7.0) and hydrogenated at room temperature for 2.5 hours. The solution was 331 mg of 10% w / w palladium on carbon.

At the end of this period, insoluble material was removed by filtration using Celite as a filtration aid and the filtrate was washed with diethyl ether. The aqueous layer was then concentrated by evaporation under reduced pressure and the residue was subjected to column chromatography using a Diaion HP-20AG column eluted with water. The eluate was concentrated by evaporation under reduced pressure and lyophilized to give the crude product. This crude product was further purified by chromatography using a Lobar column (Merck Co., LiChroprep RP-8, size B) eluting with 5% v / v aqueous methanol. The eluate was concentrated by evaporation under reduced pressure, and the residue was lyophilized to give 91 mg of the title band as a combined colorless powder. It is a mixture of two isomers (relative to the carbon atom in the 4-position of the pyrrolidine ring) in a ratio of about 1: 1.

Ultraviolet absorption spectrum (H 2 O) λmax nm: 20,300.

Nuclear Magnetic Resonance Spectrum (D 2 O, 270 MHz) δ ppm: 1.02 & 1.03 (3H, doublet x 2, J <7.33 & 6.96 Hz); 1.10 (3H, doublet, J = 6.60 Hz); 2.12 & 2.22 (1H, doublet of doublets x 2, J = 17.59 & 4.40 Hz and 17.59 & 4.03 Hz); 2.74 & 2.77 (1H, doublet of doublets x 2, J => 17.59 & 9.16 Hz and 17.59 & 9.16 Hz); 3.07 - 3.29 (3H, multiplet); 3.64-3.73 (1H, multiplet); 3.84-3.96 (1H, multiplet); 4.00 - 4.12 (2H, multiplet).

Il 35 92487

Example 3 5-Methyl-2-oxo-1,3-dioxolene-4-methylmethyl- (1R, 5S, 6S) -2- (2-oxo-3-pyrrolidinyl) -6 - [(1R) -1-hydroxyethyl] 1-Methyl-1-carbapen-2-5 eme-3-carboxylate

A solution of 66 mg of 5-methyl-2-oxo-1,3-dioxolen-4-ylmethyl iodide (prepared by heating 5-methyl-2-oxo-1,3-dioxolen-4-ylmethyl bromide and sodium iodide in acetone at reflux) 10 en) in chloroform was added to a mixture of sodium (1R, 5S, 6S) -2- (2-oxo-3-pyrrolidinylthio) -6 - [(1R) -1-hydroxyethyl] -1-methyl-1- carbapen-2-eme-3-carboxylate in 0.4 ml of dry dimethylformamide. The mixture was stirred at room temperature for 80 minutes and then at 30-45 ° C for 4.5 hours. At the end of this period, the reaction mixture was diluted with ethyl acetate and washed four times with aqueous sodium chloride solution. The extract was dried, and the solvent was removed by distillation under reduced pressure. The residue was purified by chromatography on a Lobar column (Merck, LiChroprep Si60, size A), diluted with a mixture of ethyl acetate and methanol (10: 1 by volume) to give 27.4 mg of the title compound as a colorless powder.

Ultraviolet absorption spectrum (H 2 O) λmax nm: 322.

• 25

Nuclear Magnetic Resonance Spectrum (hexadeuterated dimethyl sulphoxide, 270 MHz) δ ppm: 1.13 (3H, doublet, J = 7.32 Hz); 1.15 (3H, doublet, J = 6.35 Hz); 30 1.84-1.98 (1H, multiplet); 2.17 (3H, singlet); 2.50-2.65 (1H, multiplet); 3.12 - 3.45 (4H, multiplet); 3.77-4.06 (2H, multiplet); 4.14 (1H, doublet of doublets, J = 9.27 & 2.44 Hz); 5.08 (2H, singlet).

36 92487

Example 4

Sodium (1R, 5S, 6S) -2- (2-oxo-4-pyrrolidinyl) -6-f (1R) -1-hydroxyethyl-1-carbanen-2-em-3-carboxylate 1.45 ml of diisopropylethylamine and 1.70 ml of diphenylphosphoryl chloride were added dropwise to an ice-cooled solution containing 2.93 g of 4-nitrobenzyl- (1R, 5R, 6S) -6 - [(1R) -1-hydroxyethyl] -1-methyl- 2-Oxo-1-carbapenam-3-carboxylate in 50 ml of dry acetonitrile, and then this mixture was stirred at 0-5 ° C for 30 minutes. To this mixture, cooled to -20 ° C, was added 1.45 ml of diisopropylethylamine and a solution of 1.35 g of 4-mercapto-2-pyrrolidinone (prepared as described in "Preparation 20"). in 1 ml of acetone-15, and then the whole mixture was stirred at 0 ° C for 3 hours, after which it was allowed to stand at the same temperature overnight. The reaction mixture was then diluted with ethyl acetate, washed twice with aqueous sodium chloride solution and filtered. The organic phase was dried over anhydrous sodium sulfate, and the solvent was distilled off under reduced pressure to give the 4-nitrobenzyl ester of the title compound as a foam. It was dissolved in 150 ml of tetrahydrofuran, and the solution was filtered to remove insoluble material. 150 ml of 0.1 M phosphate buffer solution (pH 7.1) was added to the filtrate, and the catalytic reduction was carried out at room temperature * for 2.5 hours in the presence of 1.5 g of 10% w / w palladium on carbon. The reaction mixture was then worked up as described in Example 34 to give 1.05 g of the title compound as a colorless powder. It is a mixture of two isomers (relative to the carbon atom in the 4-position of the pyrrolidine ring) of about 30 in a ratio of 9: 1.

Ultraviolet absorption spectrum (H 2 O) λmax nm: 299.

35 Infrared absorption spectrum (KBr) umax cm -1: 1748, 1689, 1597, 1393, 1296.

I! 92487 37

Nuclear Magnetic Resonance Spectrum (D 2 O, 270 MHz) δ ppm: 1.03 (3H, doublet, J = 7.3 Hz); 1.10 (3H, doublet, J = 6.2 Hz); 2.12 (0.9 H, doublet of doublets, J = 17.9 & 4.4 Hz); Δ 2.22 (0.1 H, doublet of doublets, J = 17.9 & 4.4 Hz); 2.74, 2.77 (1H, 2 x doublet of doublets, J = 17.9 & 8.4 Hz); 3.08 - 3.24 (2H, multiplet); 3.26 (1H, doublet of doublets, J = 5.9 & 2.6 Hz); 3.69 (1H, doublet of doublets, J = 11.4 & 6.6 Hz); 3.84-3.93 (1H, multiplet); 4.02 - 4.11 (2H, multiplet).

Example 5 15

Sodium (1R, 5S, 6S) -2- (2-oxo-4-pyrrolidinyl) -6 - [(1R) -1-hydroxy-ethyl] -1-methyl-1-carbapen-2-em-3-yl carboxylate

The p-nitrobenzyl of the title compound was prepared as a crude product essentially following the procedure described in Example 36 above, but using 330 mg of 4-nitrobenzyl- (1R, 5R, 6S) -6 - [(IR) -1-hydroxyethyl] - l-methyl-2-oxo-l-carbapenam-3-carboxylate. This ester was subjected to column chromatography using a column containing 50 g of silica gel, eluting with a mixture of ethyl acetate and methanol (4: 1 by volume). The eluent was concentrated by evaporation under reduced pressure to give 260 mg of a colorless powder which was stirred in 10 ml of ethyl acetate. This mixture was cooled in ice and filtered to remove insoluble material. The filtrate was concentrated by evaporation under reduced pressure, and diisopropyl ether was added to the residue to give a precipitate. It was collected by filtration and dried to give 150 mg of the p-nitrobenzyl ester of the title compound as a powder. This ester is either the R or S isomer with respect to the carbon atom in the 4-position of the pyrrolidine ring.

35 38 92487

Single magnetic resonance spectrum (hexadeuterated dimethyl sulfoxide, 270 MHz) δ ppm: 1.16 (3H, doublet, J = 6.0 Hz); 1.18 (3H, doublet, J = 7.3 Hz); Δ 2.02 (1H, doublet of doublets, J = 17.1 & 4.9 Hz); 2.72 (1H, doublet of doublets, J = 17.1 & 8.3 Hz); 3.12 - 3.48 (3H, multiplet); 3.74 (1H, doublet of doublets, J = 10.7 & 6.3 Hz); 3.94-4.05 (2H, multiplet); 4.24 (1H, doublet of doublets, J = 9.8 & 2.9 Hz); 5.06 (1H, doublet, J = 4.9 Hz); 5.30, 5.46 (2H, AB, J = 14.2 Hz); 7.71 (2H, doublet, J = 8.8 Hz); 8.23 (2H, doublet, J = 8.8 Hz).

100 mg of the p-nitrobenzyl ester obtained in the above step was catalytically hydrogenated as described in Example 34 to give 55 mg of the title compound having either the R or S configuration with respect to the carbon atom at the 4-position of the pyrrolidine ring as a pure powder.

Ultraviolet absorption spectrum (H 2 O) λmax nm: 299.

Nuclear Magnetic Resonance Spectrum (D 2 O, 270 MHz) δ ppm: 1.03 (3H, doublet, J = 7.3 Hz); 1.10 (3H, doublet, J = 6.2 Hz); 2.12 (1H, doublet of doublets, J = 17.9 & 4.4 Hz); 2.74 (1H, doublet of doublets, J = 17.9 & 8.4 Hz); Δ 3.08-3.24 (2H, multiplet); 3.26 (1H, doublet of doublets, J = 5.9 & 2.6 Hz); 3.69 (1H, doublet of doublets, J = 11.4 & 6.6 Hz); 3.84-3.93 (1H, multiplet); 4.02 - 4.11 (2H, multiplet).

35

II

39 92487

Example 6

Sodium (1R, 5S, 6S) -2- (2-oxo-4-pyrrolidinylthio) -6 - [(1R) -1-hydroxyethyl] -1-methyl-1-carbapen-2-em-3-carboxylate 5

The p-nitrobenzyl of the title compound was prepared as a crude product essentially following the procedure described in Example 34 above, but using 300 mg of 4-nitrobenzyl- (1R, 5R, 6S) -6 - [(1R) -1-hydroxyethyl] -1 -methyl 2-oxo-1-carbapenam-10 3-carboxylate. This ester was subjected to column chromatography using a column containing 50 g of silica gel, eluting with a mixture of ethyl acetate and methanol (4: 1 by volume). The eluent was concentrated by evaporation under reduced pressure to give 270 mg of a colorless powder. This product is a mixture of two isomers (relative to the carbon atom in the 4-position of the pyrrolidine-15 ring) in a ratio of about 1: 1. 100 mg of this product was mixed with 10 ml of ethyl acetate. This mixture was filtered to recover insoluble material, and then this material was recrystallized from a mixture of methanol and isopropanol to give 30 mg of the p-nitrobenzyl ester of the title product as colorless needles. This ester is either the R or S isomer with respect to the carbon atom in the 4-position of the pyrrolidine ring and is the second isomer having the opposite configuration to the compound obtained in the first step of Example 37.

; 25

Nuclear Magnetic Resonance Spectrum (hexadeuterated dimethyl sulphoxide, 270 MHz) δ ppm: 1.16 (3H, doublet, J = 6.3 Hz); 1.17 (3H, doublet, J = 7.3 Hz); 2.13 (1H, doublet of doublets, J = 17.1 & 4.4 Hz); . 2.79 (1H, doublet of doublets, J = 17.1 & 7.8 Hz); 3.10 (1H, doublet of doublets, J = 10.8, 3.4 Hz); 3.16-3.35 (1H, multiplet); 3.40-3.51 (1H, multiplet); 35 3.70 (1H, doublet of doublets, J = 10.7 & 7.3 Hz); 3.95 - 4.12 (2H, multiplet); ) 40 92487 4.25 (1H, doublet of doublets, J = 9.3 & 2.5 Hz); 5.07 (1H, doublet, J = 5.4 Hz); 5.30, 5.46 (2H, AB, J = 14.2 Hz); 7.71 (2H, doublet, J = 8.8 Hz); Δ 8.23 (2H, doublet, J = 8.8 Hz).

20 mg of the p-nitrobenzyl ester obtained in the above step was catalytically hydrogenated as described in Example 34 to give 11 mg of the title compound as a pure powder having either the R or S configuration with respect to the carbon atom in the 4-position of the pyrrolidine ring, and has the opposite configuration to the compound obtained in Example 37.

Nuclear Magnetic Resonance Spectrum (D 2 O, 270 MHz) δ ppm: 1.02 (3H, doublet, J = 7.3 Hz); 1.10 (3H, doublet, J = 6.6 Hz); 2.22 (1H, doublet of doublets, J = 17.6 & 4.4 Hz); 2.77 (1H, doublet of doublets, J = 17.6 & 8.4 Hz); 3.08 - 3.25 (2H, multiplet); 3.25 (1H, doublet of doublets, J = 5.9 & 2.6 Hz); 3.68 (1H, doublet of doublets, J = 11.4 & 6.4 Hz); 3.84-3.96 (1H, multiplet); 4.00 - 4.12 (2H, multiplet).

25 Example 7

Pivaloyloxymethyl (IR.5S.6S) -2- (2-oxo-4-pyrrolidinyl) -6-f (IR) -1-hydroxyethyl 11-methyl-1-carbapen-2-em-3-carboxylate 30 100 mg in an example Sodium 2- (2-oxo-4-, pyrrolidinylthio) -6 - [(IR) -1-hydroxyethyl] -1-methyl-1-carbapen-2-eme-3-carboxylate obtained as described 36 was suspended in 3 ml: dry N, N-dimethylacetamide. To this suspension, 80 μΐ of pivaloyloxymethyl iodide was added under ice-cooling, and then the mixture was stirred for 15 to 35 minutes, during which time the mixture became a solution. The reaction mixture was then diluted with 50 ml of ethyl acetate and washed twice with aqueous sodium chloride solution. The organic phase was dried over anhydrous sodium sulfate, and the solvent was removed by distillation under reduced pressure. The residue was purified by chromatography on a Lobar column (Merck, LiChroprep RP-8, size B) eluting with 60% v / v aqueous methanol. The eluent was evaporated under reduced pressure to remove methanol, leaving an aqueous phase which was stirred with sodium chloride and then extracted with ethyl acetate. The extract was dried over anhydrous sodium sulfate and concentrated by evaporation under reduced pressure to give 110 mg of the title compound as a colorless powder.

Ultraviolet absorption spectrum (CH 3 CN) λmax nm: 323 (€ = 10760).

Infrared absorption spectrum (KBr) vmax cm'1: 1778, 1756, 1699.

Nuclear Magnetic Resonance Spectrum (CDCl 3, 270 MHz) δ ppm: 1.23 (9H, singlet); 1.29 (3H, doublet, J = 7.3 Hz); 1.35 (3H, doublet, J = 6.2 Hz); 1.95 (1H, broad singlet); 2.33 (1H, doublet of doublets, J = 17.6 & 6.2 Hz); 2.79 (1H, doublet of doublets, J = 17.6 & 8.8 Hz); 3.22 - 3.34 (2H, multiplet); 3.38 (1H, doublet of doublets, J * = 9.9 & 4.8 Hz); 3.80 (1H, doublet of doublets, J = 10.3 & 7.0 Hz); 3.95-4.05 (1H, multiplet); 4.20 - 4.27 (2H, multiplet); 5.79 (1H, broad singlet); 5.83, 5.97 (2H, AB, J = 5.5 Hz).

42 92487

Example 8 (1-Methylcyclohexane-1-methylcarbonyloxymethyl- (1R, 5S, 6S) -2- (2-oxo-4-pyrrolidinylmethyl) -6 - [(1R) -1-hydroxyethyl] -1-methyl-1 -5-Carbapen-2-eme-3-carboxylate This example followed a procedure similar to that described in Example 39, but using (1-methylcyclohexan-1-yl) carbonyloxymethyl iodide instead of pivaloyloxymethyl iodide to give the title compound.

Ultraviolet Absorption Spectrum (CH 3 CN) λmax nm: 323 (ε-9655).

Infrared absorption spectrum (KBr) umax cm -1: 1777, 1753, 1700.

Nuclear Magnetic Resonance Spectrum (CDCl 3, 270 MHz) δ ppm: 1.18 (3H, singlet); 1.29 (3H, doublet, J = 7.3 Hz); 1.35 (3H, doublet, J = 6.2 Hz); 1.25-1.75 (9H, multiplet); 2.00 - 2.08 (2H, multiplet); 2.33 (1H, doublet of doublets, J = 17.6 & 6.2 Hz); 2.79 (1H, doublet of doublets, J = 17.6 & 8.8 Hz); 3.21 - 3.33 (2H, multiplet); 3.38 (1H, doublet of doublets, J = 9.9 & 4.8 Hz); 3.79 (1H, doublet of doublets, J = 9.9 & 7.0 Hz); 3.95-4.05 (1H, multiplet); Δ 4.21-4.30 (2H, multiplet); 5.78 (1H, broad singlet); 5.87, 5.96 (2H, AB, J = 5.5 Hz).

IJ

1- (cyclohexyloxycarbonyl) ethyl - (1R, 5S, 6S) -2- (2-oxo-4-pyrrolidinylthio) -6 - [(1R) -1-hydroxyethyl] -1-methyl-1-carbapen-2- eme-3-carboxylate 43,92487

Example 9 This example followed a procedure similar to that described in Example 39 using 54 g of sodium 2- (2-oxo-4-pyrrolidinylthio) -6 - [(1R) -β-hydroxyethyl] obtained by a method similar to that described in Example 36. 1-methyl-1-carbapen-2-em-3-carboxylate and 50 μΐ of 1- (cyclohexyloxycarbonyloxy) ethyl iodide to give 62 mg of the title compound as a colorless powder.

Ultraviolet Absorption Spectrum (CH3CN) Amax run: 15 323 (e - 10766).

Infrared Absorption Spectrum (KBr) umax cm'1: 1759, 1701.

Nuclear Magnetic Resonance Spectrum (CDCl 3, 270 MHz) δ ppm: 1.28 (3H, doublet, J = 7.0 Hz); 1.33, 1.36 (3H, two doublets, J = 6.2 Hz); 1.59, 1.61 (3H, two doublets, J = 5.5 Hz); 1.2-1.6 (6H, multiplet); : Δ 1.7-2.0 (5H, multiplet); 2.33 (1H, doublet of doublets, J = 17.6 & 6.2 Hz); 2.79 (1H, doublet of doublets, J = 17.6 & 8.8 Hz); 3.24 - 3.33 (2H, multiplet); 3.35-3.42 (1H, multiplet); 3.75-3.85 (1H, multiplet); * 3.94-4.05 (1H, multiplet); 4.19 - 4.27 (2H, multiplet); 4.59-4.70 (1H, multiplet); 5.69 (1H, broad singlet); 35 6.88 (1H, quartet, J = 5.5 Hz).

44 92487

Example 10 1- (Cyclopentylloxycarbonyl) ethyl (C 1 -C 5 S 6 S) -2- (2-oxo-4-pyrrolidinylthio) -6-i (1R) -1-hydroxyethyl-1-methyl-1-carbapen-2- Eem-3-carboxylate In this example, a procedure similar to that described in Example 39 was followed, except that 1- (cyclopentyloxycarbonyloxy) ethyl iodide was used to give the title compound.

10

Ultraviolet Absorption Spectrum (CH 3 CN) λmax nm: 323 (e - 10651).

Infrared absorption spectrum (KBr) umax cm'1: 15 1760, 1701.

Nuclear Magnetic Resonance Spectrum (CDCl 3, 270 MHz) δ ppm: 1.28, 1.29 (3H, two doublets, J = 7.3 Hz); 1.33, 1.35 (3H, two doublets, J = 6.2 Hz); 1.58, 1.60 (3H, two doublets, J = 5.5 Hz); 1.65-1.95 (8H, multiplet); 2.33 (1H, doublet of doublets, J = 17.6 & 6.2 Hz); 2.79 (1H, doublet of doublets, J = 17.6 & 8.8 Hz); 3.23 - 3.32 (2H, multiplet); • j 3.38, 3.39 (1H, two doublets of doublets, J = 9.9 & 4.8

Hz); 3.68-3.87 (1H, multiplet); 3.96-4.07 (1H, multiplet); 4.19 - 4.27 (2H, multiplet); 5.08-5.16 (1H, multiplet); 5.66 (1H, broad singlet); 6.87 (1H, quartet, J = 5.5 Hz).

II

1- (cyclohexylmethyloxycarbonylloxy) methyl- (1R, 5S, 6S-) -2- (R-oxo-N-pyrrolidinylthiol-6-β-ClR) -1-hydroxyethyl-1-methyl-1-5-carbapen-2-em -3-carboxylate 45,92487

Example 11 This example followed a procedure similar to that described in Example 39, but using 1- (cyclohexylmethyloxycarbonyloxy) ethyl iodide to give the title compound.

Ultraviolet absorption spectrum (CH 3 CN) λmax nm: 323 (e = 10975).

Infrared absorption spectrum (KBr) umax cm -1: 1766, 1700, 1269.

Nuclear Magnetic Resonance Spectrum (CDCl 3, 270 MHz) δ ppm: 0.95-1.26 (5H, multiplet); 1.28, 1.29 (3H, two doublets, J = 7.3 Hz); 1.33, 1.35 (3H, two doublets, J = 6.2 Hz); 1.59, 1.61 (3H, two doublets, J = 5.9 & 5.5 Hz); 1.64 - 1.76 (6H, multiplet); 1.87 (1H, broad singlet); • 25 2.33 (1H, doublet of doublets, J = 17.6 & 6.2 Hz); 2.79 (1H, doublet of doublets, J = 17.6 & 8.8 Hz); 3.24 - 3.35 (2H, multiplet); 3.37-3.42 (1H, multiplet); 3.75-3.87 (1H, multiplet); Δ 3.95-4.06 (3H, multiplet); 4.20 - 4.28 (2H, multiplet); 5.73 (1H, broad singlet); 6.86, 6.87 (1H, quartet, J = 5.5 Hz).

35

Example 12 92487 46 1- (Isopropoxycarbonyloxy-ethyl- (1R.55.63) -2- (2-oxo-4-pyrrolidinylthio) -6 - [(1R) -1-hydroxyethyl] -1-methyl-1-carbapen -2-Eemi-3-5 Carboxylate In this example, a procedure similar to that described in Example 39 was followed, but using 1- (isopropoxycarbonyloxy) ethyl iodide to give the title compound.

10

Ultraviolet Absorption Spectrum (CH 3 CN) λmax nm: 323 (e - 10961).

Infrared absorption spectrum (KBr) vmax cm'1: 1762, 1701, 1272.

Nuclear Magnetic Resonance Spectrum (CDCl 3, 270 MHz) δ ppm: 1.26-1.36 (12H, multiplet); 1.39, 1.61 (3H, two doublets, J = 5.5 Hz); 1.80 (1H, broad triplet, J = 3.7 Hz); 2.33 (1H, doublet of doublets, J = 17.6 & 6.2 Hz); 2.79 (1H, doublet of doublets, J * = 17.6 & 8.8 Hz); 3.24 - 3.34 (2H, multiplet); 3.35-3.42 (1H, multiplet); ·· 25 3.76-3.87 (1H, multiplet); 3.95-4.07 (1H, multiplet); 4.19-4.28 (2H, multiplet); 4.82-4.99 (1H, multiplet); 5.63 (1H, broad singlet); 6.88 (1H, quartet, J = 5.5 Hz).

(R) 5S, 6S) - (1) -methyloxycarbonylmethoxymethyl- (1R, 5S, S, -2- (2-oxo-4-pyrrolidinylthio) -6-f (IR) -1-hydroxyethyl) 11-methyl- 1-carbapen-2-ene-3-carboxylate 47,92487

Example 13 This example followed a procedure similar to that described in Example 39, but using (1R, 2S, 5R) - (1) -methyloxycarbonyloxymethyl iodide to give the title compound.

10

Ultraviolet absorption spectrum (CH 3 CN) λmax nm: 324 (e = 10801).

Infrared absorption spectrum (KBr) umax cm'1: 15 1763, 1695, 1266.

Nuclear Magnetic Resonance Spectrum (CDCl 3, 270 MHz) δ ppm: 0.78 (3H, doublet, J = 7.0 Hz); 0.89 (3H, doublet, J = 7.0 Hz); 0.92 (3H, doublet, J = 7.3 Hz); 1.29 (3H, doublet, J = 7.3 Hz); 1.35 (3H, doublet, J = 6.2 Hz); 0.95-1.55 (5H, multiplet); 1.63 - 1.73 (2H, multiplet); i 1.88-2.03 (1H, multiplet); 2.08-2.17 (1H, multiplet); 2.34 (1H, doublet of doublets, J = 17.2 & 6.2 Hz); 2.79 (1H, doublet of doublets, J = 17.2 & 8.8 Hz); 3.22 - 3.34 (2H, multiplet); 3.39 (1H, doublet of doublets, J = 9.5 & 4.8 Hz); 3.74-3.86 (1H, multiplet); 3.96-4.06 (1H, multiplet); 4.18-4.28 (2H, multiplet); 4.56 (1H, doublet of triplets, J = 10.6 & 4.4 Hz); 35 5.70 (1H, broad singlet); 5.89, 5.91 (2H, AB, J = 5.9 Hz).

48 92487

Example 14 5-Methyl-2-oxo-1. 3-Dioxolen-4-methylmethyl- (1R, 5S, 6S) -2- (2-oxo-4-pyrrolidinylthio) -6 - [(1R) -1-hydroxyethyl] -1-methyl-1-carbapen-2-5 eem-3-carboxylate In this example, a procedure similar to that described in Example 39 was followed, but using 50 mg of sodium 2- (2-oxo-4-pyrrolidinylthio) -6 - [(IR) -1-hydroxyethyl] -1 methyl 1-carbapen-2-ene-3-10 carboxylate and 55 mg of 5-methyl-2-oxo-1,3-dioxolen-4-ylmethyl bromide to give 50 mg of the title compound as a colorless powder.

Ultraviolet Absorption Spectrum (CH 3 CN) Amax run: 15 321.1 (f - 8982).

Infrared absorption spectrum (KBr) vmax cm -1: 1: 1820, 1772, 1701, 1627.

Nuclear Magnetic Resonance Spectrum (CDCl 3, 270 MHz) δ ppm: 1.30 (3H, doublet, J = 7.33 Hz); 1.36 (3H, doublet, J = 6.22 Hz); 1.80 (1H, broad singlet); 2.21 (3H, singlet); : 25 2.35 (1H, doublet of doublets, J = 17.58 & 6.04 Hz); 2.81 (1H, doublet of doublets, J = 17.58 & 8.80 Hz); 3.25 - 3.43 (3H, multiplet); 3.73 - 4.05 (2H, multiplet); 4.17-4.32 (2H, multiplet); 4.96, 5.05 (2H, AB, J = 13.93 Hz); 5.59 (1H, broad singlet.

Example 15 49 92487

Examples 15-24 5 1- (Isobutryloxy) ethyl- (1R, 5S, 6S) -2- (2-oxo-4-pyrrolidinyl) -1- (1R) -1-hydroxyethyl-1-methyl-1-carbapen -2-em-3-carboxylic-methacrylate

Ultraviolet Absorption Spectrum (CH 3 CN) λmax nm: 10,323.

Example 16 1- (Difluorohexanecarbonyloxy) ethyl - (1R, 5S, 6S) -2- (2-oxo-4-pyrrole-15-lidinyl) -6-f (IR) -hydroxyethyl-1-methyl-1-carbapentene 2-em-3-carboxylate

Ultraviolet Absorption Spectrum (CH3CN) λmax nm: 323.

20

Example 17 1- (Pivaloyloxy) ethyl- (1R, 5S, 6S-) -2- (7-oxo-4-pyrrolidinyl) -6-yl (1R) -1-hydroxyethyl-1-methyl-1-carboxyl-2 -em-3-carboxylate: 25

Ultraviolet Absorption Spectrum (CH3CN) λmax nm: 323.

Example 18 1-Acetoxyethyl - (1R, 5S, 6S ') - 2- (2-oxo-4-pyrrolidinyl) -6 - [(1R) -1-hydroxyethyl] -1-methyl-1-carbapen-2- em-3-carboxylate

Ultraviolet Absorption Spectrum (CH 3 CN) λmax nm: 35,323.

Example 19 50 92487

Cyclohexanecarbonylloxyethyl (1R, 5S, 6S, 2- (2-oxo-4-pyrrolidinyl) thio] -6 - [(1R) -1-hydroxyethyl] -1-methyl-1-carbapen-2-em-3 -5 carboxylate

Ultraviolet Absorption Spectrum (CH3CN) λmax nm: 323.

10 Example 20

Cyclohexylloxycarbonylmethoxymethyl- (1R, 5S, 6S) -2- (2-oxo-4-pyrrolidinylthio) -6 - [(1R) -1-hydroxyethyl] -1-methyl-1-carbapen-2-em-3- carboxylate 15

Ultraviolet Absorption Spectrum (CH3CN) λmax nm: 323.

Example 21 20

Cyclopentyloxycarbonylmethoxymethyl- (1R, 5S-es) -2- (2-oxo-4-pyrrolidinyl) -6 - [(1R) -1-hydroxyethyl] -1-methyl-1-carbapen-2-emetic 3-carboxylate: Ultraviolet absorption spectrum (CH3CN) λmax nm: 323.

Example 22 Sodium (1R, 5S, 6S) -2- (1-methyl-2-oxo-4-pyrrolidinyl), -6 - [(1R) -1-hydroxy] -1-methyl-1 -carbapen-2-emem-3-carboxylate

Ultraviolet Absorption Spectrum (CH 3 CN) λmax nm: 300.

35 ii 92487 51

Example 23

Pivaloyloxymethyl- (1R, 5S, 6S) -2- (1-methyl-2-oxo-4-pyrrolidin-lithio) -6 - [(1R) -1-hydroxyethyl-1-methyl-1-carbapen-2- eem-3-carbox-5 slate

Ultraviolet Absorption Spectrum (CH3CN) λmax nm: 323.

Example 24 1- (Isoporpoxycarbonyloxy-ethyl-5S, 6S) -2- (1-methyl-2-oxo-4-pyrrolidinylthio-4 - [(1R) -1-hydroxyethyl] -1-methyl-1-methyl) -1- carbapen-2-eme-3-carboxylate 15

Ultraviolet Absorption Spectrum (CH3CN) λmax nm: 323.

Preparation 1 20 (2S.45) -2-Carbamoyl-4- (4-methoxybenzylthio) -1,1-dimethylpyrrolidinium fluorosulfonate 1- (1) (2S.4R) -lt-butoxycarbonyl) -4-hydroxy-2- pyrrolidinecarboxylic acid 27.3 g of sodium hydroxide was dissolved in 380 ml of water, and 85 g of (2S, 4R) -4-hydroxy-2-pyrrolidinecarboxylic acid were added to this solution at 3-5 ° C, and then at the same temperature 570 ml of tetrahydrofuran. A solution of 141.5 g of di-t-butoxycarbonate in 190 ml of tetrahydrofuran was added to the reaction mixture at 3-5 ° C, and then the mixture was stirred at 50-55 ° C for 2 hours. At the end of this period, the mixture was cooled and then adjusted to pH 3-4 by the addition of concentrated hydrochloric acid and then ammonium chloride. The mixture was then extracted with tetrahydrofuran, and the extract was dried. The solvent was then removed from 52,92487 of the extract by distillation under reduced pressure to give 128 g of the title compound as a colorless powder.

Nuclear Magnetic Resonance Spectrum (CDCl 3, 60 MHz) δ ppm: δ 1.44 (9H, singlet); 1.96 - 2.45 (2H, multiplet); 2.36 - 2.72 (2H, multiplet); 4.24 - 4.66 (2H, multiplet); 5.04-5.60 (2H, broad singlet).

1- (2) (2S, 4R) -lt-butoxycarbonyl-2-carbamoyl-4-hydroxypyrrolidine 58 g of (2S, 4R) -1- (t-butoxycarbonyl) -4-hydroxy-2-pyrrolidinecarbox-15 the silicic acid [prepared as described in step (1) above] was dissolved in 850 ml of dry tetrahydrofuran, and then 38.2 ml of triethylamine was added to this mixture at a temperature of -15 to -20 ° C. A solution of 26.3 ml of ethyl chloroformate in 240 ml of dry tetrahydrofuran was then added dropwise to this mixture at -15 to -20 ° C, and then the mixture was stirred at this same temperature for 35 minutes. At the end of this period, 258 ml of a 28 w / v% aqueous solution of ammonium hydroxide was added to the mixture at -15 to -20 ° C, and the mixture was allowed to stand overnight at room temperature.

Ammonium chloride was then added to the reaction mixture and extracted: then with tetrahydrofuran. The extract was dried, and the solvent was removed by distillation under reduced pressure. The resulting residue was triturated with diethyl ether to initiate crystallization. The crystals were collected by filtration and washed with diethyl ether to give 49.7 g of the title compound as colorless crystals, m.p. 146-148 ° C.

Il 92487 53

Nuclear Magnetic Resonance Spectrum (hexadeuterated dimethyl sulphoxide, 60 MHz) δ ppm: 1.38 (9H, singlet); 1.65 - 2.24 (2H, multiplet); Δ 3.00-3.66 (2H, multiplet); 3.76-4.49 (3H, multiplet); 6.78 (1H, broad singlet); 7.23 (1H, broad singlet).

1- (3) (2S, 4R) -1- (t-butoxycarbonyl) -2-carbamoyl-4-methanesulfonoxyphyrrolidine 1.85 ml of methanesulfonyl chloride was added to a cooled solution of 5.0 g (2S. 4R) -1- (t-butoxycarbonyl) -2-carba-15-moyl-4-hydroxypyrrolidine [prepared as described in step (2) above] in 250 ml of dry tetrahydrofuran, to which was added 3.31 ml of triethylamine. The mixture was then stirred at 0-5 ° C for 1 hour, after which it was poured into aqueous sodium chloride solution and extracted with ethyl acetate.

The extract was washed with aqueous sodium chloride solution and dried over anhydrous magnesium sulfate, and the solvent was removed by distillation under reduced pressure. The residue was purified by chromatography on silica gel eluting with a mixture of ethyl acetate and methanol (9: 1 by volume) to give 5.5 g of the title compound as colorless crystals.

Nuclear Magnetic Resonance Spectrum (CDCl 3, 60 MHz) δ ppm: 1.43 (9H, singlet); 2.10 - 2.68 (2H, multiplet); 3.12 (3H, singlet); 3.10-3.40 (1H, broad singlet); 3.73 (2H, doublet, J = 4.0 Hz); 4.32 (1H, triplet, J = 7.0 Hz); 5.26 (1H, triplet, J = 4.0 Hz); 35 6.68 (1H, broad singlet); 7.30 (1H, broad singlet).

54 92487 1- (41) (2S, 4R) -1- (t-butoxycarbonyl) -2-carbamoyl-4- (4-methoxybenzyl) pyrrolidine 330 mg of sodium hydride (55 w / w as a dispersion in mineral-5 oil) ) was added under ice-cooling to a solution of 1.18 g of p-methoxybenzyl mercaptan in 25 ml of dry dimethylformamide, and then the mixture was stirred at room temperature for 30 minutes.

At the end of this time, 2.14 g of (2S, 4R) -1- (t-butoxycarbonyl) -2-carbamoyl-4-methanesulfonyloxypyrrolidine [prepared as described in step (3) above] was added to the mixture, and then the mixture was stirred. at room temperature for 3 hours. The reaction mixture was then poured into aqueous sodium chloride solution and extracted with ethyl acetate. The extract was washed with aqueous sodium chloride solution and dried over anhydrous magnesium sulfate, and the solvent was removed by distillation under reduced pressure. The residue was purified by chromatography on silica gel eluting with a mixture of cyclohexane and ethyl acetate (2: 3 by volume) to give 1.94 g of the title compound as an oil.

Nuclear Magnetic Resonance Spectrum (CDCl 3, 60 MHz) & ppm: 1.43 (9H, singlet); 1.80 - 3.42 (5H, multiplet); 3.70 (2H, singlet); 3.78 (3H, singlet); 4.18 (1H, triplet, J = 7.0 Hz); 5.96 (1H, broad singlet); 6.35 (1H, broad singlet); 6.79, 7.21 (4H, A2B2, J = 9.0 Hz).

1- (5) (2S, 4S) -2-carbamovyl-4- (4-methoxybenzyl) pyrrolidine 1.92 g of (2S, 4S) -1- (t-butoxycarbonyl) -2-carbamoyl-4- (4 -methoxy-benzylthio) pyrrolidine [prepared as described in step (4) above] was dissolved in 25 ml of ethyl acetate, and to this solution, which was cooled in ice, 26.2 ml of a 4N dioxane solution of hydrogen chloride was added. The mixture was then stirred at 0-5 ° C! 92487 55 2 hours and at room temperature for 30 minutes. At the end of this period, it was poured into a saturated aqueous solution of sodium bicarbonate to make it weakly acidic and then extracted with ethyl acetate. The extract was washed with aqueous sodium chloride solution and dried over anhydrous magnesium sulfate. The solvent was then removed by distillation under reduced pressure to give 1.36 g of the title compound as a powder having a melting point of 120-121 ° C.

Nuclear Magnetic Resonance Spectrum (hexadeuterated dimethyl sulphoxide, 60 MHz) δ ppm: 1.58 - 3.40 (7H, multiplet); 1.67 (2H, singlet); 1.78 (3H, singlet); 6.43 (1H, broad singlet); 6.78 (7.22 (4H, A2B2, J = 9.0 Hz); 7.31 (1H, broad singlet).

1- (5a) (2S, 4S) -2-Carbamoyl-4- (4-methoxybenzyl) pyrrolidine hydrochloride 620 ml of a 4N solution of hydrogen chloride in ethyl acetate were added to an ice-cooled solution containing 91 g of (2S, 4S) -1 -butoxycarbonyl-2-carbamoyl-4- (4-methoxybenzylthio) pyrrolidine [prepared as described in step (4) above] in 2 liters of ethyl acetate; 25 tats, and then this mixture was stirred at room temperature for 4.5 hours. The precipitated crystals were collected by filtration, washed with diethyl ether and dried in vacuo to give 63 g of the title compound having a melting point of 192-195 ° C.

Infrared absorption spectrum (KBr) umax cm-1: 1706, 1584, 1512.

56 92487

Nuclear Magnetic Resonance Spectrum (D 2 O, 270 MHz) δ ppm: 1.75-1.90 (1H, multiplet); 2.54-2.62 (1H, multiplet); 3.04-3.11 (1H, multiplet); Δ 3.28-3.41 (2H, multiplet); 3.64 (2H, singlet); 3.65 (3H, singlet); 4.22 (1H, triplet, J = 8.06 Hz); 6.80, 7.15 (4H; A2B2, J = 8.79 Hz).

10- (6a) (2S, 4S) -2-carbamoyl-4- (4-methoxybenzyl) -1-methylpyrrolidine

A solution of 0.29 g of sodium bicarbonate in 3 ml of water and 0.34 ml of 35% formalin was added dropwise, in this order, to an ice-cooled suspension containing 1 g of the compound obtained in step 1- (5a) above in 20 ml. acetonitrile, and then the mixture was stirred at 10 ° C for 20 minutes. To this mixture, which was cooled in ice, was then added 0.25 g of sodium cyano-20 borohydride, and the mixture was stirred at room temperature for 20 minutes. To this mixture, 0.5 ml of acetic acid was added under ice-cooling, and then it was stirred at room temperature for 20 minutes. At the end of this period, the mixture was diluted with 100 ml of ethyl acetate and washed successively with 1N aqueous sodium hydroxide solution and sodium hydroxide solution. 25 with a mixture of saturated aqueous sodium chloride solution (1: 1 by volume) and then with saturated aqueous sodium chloride solution. The organic phase was then dried over anhydrous magnesium sulfate, and the solvent was removed by distillation under reduced pressure. The residue was purified by column chromatography on silica gel (Wakogel C-100, Wako Junyaku) eluting with a mixture of chloroform and methanol (v / v • # 95: 5) to give 0.92 g of the title compound. The melting point, infrared absorption spectrum and nuclear magnetic resonance spectrum of this compound were the same as those of the compound obtained as described in 1- (6) below.

35

II

92487 57 1- (6) (2S, 45) -2-carbamoyl-4- (4-methoxybenzylthio) -1-methylpyrrolidine 0.07 ml of methyl iodide was added to a cooled solution of 0.6 g (2S, 4S) -2-Carbamoyl-4- (4-methoxybenzylthio) pyrrolidine (prepared as described in step (5) above) dissolved in 4.5 ml of dry dimethylformamide This mixture was then stirred at 0-5 ° C. 5 minutes and at room temperature for 20 minutes At the end of this time, the reaction mixture was poured into saturated aqueous sodium bicarbonate solution and extracted with ethyl acetate. chromatography on a Lobar column (Merck, LiChroprep Si60, size B) eluting with a mixture of ethyl acetate and methanol (9: 1 v / v) to give 252 mg of the title compound. crystals with a melting point of 113-114 ° C.

Infrared absorption spectrum (KBr) uraax cm-1: 20 1636, 1609, 1512.

Nuclear Magnetic Resonance Spectrum (CDCl 3, 60 MHz) δ ppm: 1.58-3.36 (6H, multiplet); 2.35 (3H, singlet); . Δ 3.68 (2H, singlet); 3.78 (3H, singlet); 5.95 (1H, broad singlet); 6.84, 7.23 (4H, A2B2, J = 9.0 Hz); 7.20 (1H, broad singlet).

30. 1- (7) (2S, 4S) -2-Carbamoyl-4- (4-methoxybenzyl) -1,1-dimethylpyrrolidinium fluorosulfonate 0.123 ml of methylsulfonate was added to an ice-cooled solution containing 320 mg of (2S, 4S). 2-Carbamoyl-4- (4-methoxybenzylthio) -1-methylpyrrolidine [prepared as described in step (6) above or 58,92487 (6a)] dissolved in 7 ml of dry methylene chloride. The mixture was then stirred at this same temperature for 20 minutes and at room temperature for 2 hours. At the end of this period, the solvent was removed by distillation under reduced pressure, and the residue 5 was washed several times with diethyl ether between decantations and then dried under reduced pressure to give 525 mg of the title compound as an oil.

Nuclear Magnetic Resonance Spectrum (D 2 O, 270 MHz) δ ppm: δ 2.01-3.68 (5H, multiplet); 3.02 (3H, singlet); 3.03 (3H, singlet); 3.65 (3H, singlet); 3.68 (2H, singlet); 4.07 (1H, doublet of doublets, J = 8.43 & 7.70 Hz); 6.81, 7.16 (4H, A2B2, J = 8.79 Hz).

Preparation 2 20 (2S, 4S) -2-Carbamoyl-4- (4-methoxybenzyl) -1- (2-fluoroethyl) -1-methylpyrrolidinium fluorosulfonate 2-Q) (2S, 4S) -2-Carbamoyl-4- (4-Methoxybenzyl) -1- (2-fluoroethyl) pyrrolidine: 25 ', 0.4 ml of 1-bromo-2-fluoroethane, 3.83 g of sodium iodide and 0.38 g of sodium bicarbonate were added. in a cooled solution of 1.2 g of (2S, 4S) -2-carbamoyl-4- (4-methoxybenzylthio) pyrrolidine dissolved in 12 ml of dry dimethylformamide, and this mixture was stirred at room temperature for 20 minutes and then • At 40 eC for 20 hours. At the end of this period, the reaction mixture was poured into saturated aqueous sodium bicarbonate solution and extracted with ethyl acetate. The extract was washed with saturated aqueous sodium chloride solution and dried over anhydrous magnesium sulfate. The solvent was then removed by distillation under reduced pressure, and the residue was subjected to column chromatography on silica gel (Ka- 92487 59 tayma Chemicals Industries Co., Silica gel 60K070). Fractions eluted with ethyl acetate gave 838 mg of the title compound as a powder, m.p. 122-123 ° C. Infrared Absorption Spectrum (KBr) umax cm -1: 1636, 1610, 1510.

Nuclear Magnetic Resonance Spectrum (CDCl 3, 60 MHz) δ ppm: 1.55 - 3.40 (8H, multiplet); Δ 3.69 (2H, singlet); 3.79 (3H, singlet); 4.47 (2H, doublet of triplets, J = 47.0 & 6.0 Hz); 4.78 (1H, broad singlet); 7.02 (4H, A2B2, J = 9.0 Hz); Δ 6.95-7.50 (1H, broad singlet).

2- (2) (2S, 4S) -2-Carbamoyl-4- (4-methoxybenzyl) -1- (2-fluoroethyl) -1-methylpyrrolidinium fluorosulfonate 0.17 ml of methyl fluorosulfonate was added to an ice-cooled solution. containing 630 mg of (2S, 4S) -2-carbamoyl-4- (4-methoxybenzylthio) -1- (2-fluoroethyl) -pyrrolidine [prepared as described in step (1) above] dissolved in 12 ml of: dry methylene chloride. The mixture was stirred at this same temperature for 30 minutes; 25 and then at room temperature for 5 hours. At the end of this period, the solvent was removed by distillation under reduced pressure, and the residue was washed several times with diethyl ether, decanted, and dried under reduced pressure to give 850 mg of the title compound as an oil.

30. Nuclear Magnetic Resonance Spectrum (D 2 O, 270 MHz) δ ppm: 1.84 - 4.73 (10 H, multiplet); 3.18 (3H, singlet); 3.65 (3H, singlet); 35 3.68 (2H, singlet); 6.79-7.19 (4H, multiplet).

92487 60

Preparation 3 (2S. 4S) -1. 1-Dimethyl-2-methylcarbanoyl-4-CA-methoxybenzylthio) -pyrrolidinium fluorosulfonate 5 3- (1) (2S, 4R ") -1- (t-butoxycarbonyl) -2-methylcarbamoyl-4-hydroxypyrrolidine 9 , 91 ml of triethylamine was added at -40 ° C to a solution of 15.03 g of (2S, 4R) -1- (t-butoxycarbonyl) -4-hydroxy-2-pyrrolidinecarboxylic acid dissolved in 250 ml of dry tetrahydric acid. rofuran, and then a solution of 6.81 ml of ethyl chloroformate in 30 ml of dry tetrahydrofuran was added to the resulting solution at -30 to 40 ° C. The reaction mixture was then stirred at this same temperature for 1 hour. 16.82 ml of a 40% strength by volume aqueous solution of methylamine were then added at -20 [deg.] C., and then the temperature was allowed to rise to room temperature, and the reaction was allowed to continue for a further hour at the end of this time. se uu was taken three times with ethyl acetate. The combined extracts were washed with aqueous sodium chloride solution and dried over anhydrous magnesium sulfate. The solvent was then removed by distillation under reduced pressure to give 12.76 g of the title compound as a colorless oil.

: 25

Nuclear Magnetic Resonance Spectrum (CDCl 3, 60 MHz) δ ppm: 1.44 (9H, singlet); 1.89 - 2.45 (2H, multiplet); 2.81 (3H, doublet, J = 5.0 Hz); 3.23 - 3.71 (3H, multiplet); : 4.12-4.68 (2H, multiplet); 6.70 (1H, broad singlet).

II

92487 61 3- (2) (2S, 4R) -1- (t-Butoxycarbonyl) -4-methanesulfonyloxy-2-methylcarbamoylpyrrolidine 7.11 mL of triethylamine was added to a cooled solution of ice containing 11.29 g (2S). -1- (t-Butoxycarbonyl) -2-methylcarbamoyl-4-hydroxypyrrolidine [prepared as described in step (1) above] dissolved in 120 ml of dry tetrahydrofuran, and 3.93 ml of methanesulfonyl chloride was added to the resulting mixture and stirred at 0 ° C. At -5 eC for 30 minutes and then at 10 15 eC for 30 minutes. The mixture was then poured into aqueous sodium chloride solution and extracted with ethyl acetate. The extract was washed with aqueous sodium chloride solution and dried over anhydrous magnesium sulfate. The solvent was then removed by distillation under reduced pressure to give 11.58 g of the title compound as color-free crystals.

Nuclear Magnetic Resonance Spectrum (CDCl 3, 60 MHz) δ ppm: 1.46 (9H, singlet); 2.00 - 2.85 (2H, multiplet); 2.81 (3H, doublet, J = 5.0 Hz); 3.03 (3H, singlet); 3.41-5.42 (4H, multiplet); 6.75 (1H, broad singlet).

. 3- (3) (2S, 4S) -1- (t-butoxycarbonyl) -4- (4-methoxybenzyl) -2-methylcarbamoylpyrrolidine 1.80 g of sodium hydride (55% w / w dispersion in mineral oil) was added in ice. cooling to a solution of 5.70 ml of 4-methoxybenzyl mercaptan dissolved in 100 ml of dry tetrahydrofuran and then the mixture was stirred at 0-5 ° C for 30 minutes. At the end of this time, a solution of 11.00 g of (2S, 4R) -1- (t-butoxycarbonyl) -4-methanesulfonyl-oxy-2-methylcarbamoylpyrrolidine [prepared as described in Step 35 (2) above] in 80 ml of dry matter was added to the mixture. dimethylformamide, and then the mixture was stirred at 34 ° C for 4.5 hours. The reaction mixture was then poured into aqueous sodium chloride solution and extracted with ethyl acetate. The extract was washed with aqueous sodium chloride solution and dried over anhydrous magnesium sulfate, and the solvent was removed by distillation under reduced pressure. The residue was purified by chromatography on silica gel eluting with a mixture of ethyl acetate and hexane (5: 1 by volume) to give 4.35 g of the title compound as an oil.

Nuclear Magnetic Resonance Spectrum (CDCl 3, 60 MHz) δ ppm: δ 1.42 (9H, singlet); 1.80 - 4.40 (6H, multiplet); 2.81 (3H, doublet, J = 5.0 Hz); 3.70 (2H, singlet); 3.79 (3H, singlet); 6.39 (1H, broad singlet); 6.87, 7.26 (4H, A2B2, J = 9.0 Hz).

3- (4) (2S, 4S) -4- (N-methoxybenzyl) -2-methylcarbamoyl pyrrolidine 52.5 ml of a 4N dioxane solution of hydrogen chloride were added to a cooled solution of 4.00 g (2S, 45>) in ice. ) -1- (t-butoxycarbonyl) -4- (4-methoxybenzylthio) -2-methylcarbamoylpyrrolidine [prepared as described in step (3) above] dissolved in 50 ml of ethyl acetate. The mixture was then stirred at 0-5 ° C for 2 hours and at room temperature for 30 minutes. At the end of this period, it was poured into saturated aqueous sodium bicarbonate solution to make it weakly acidic, and then extracted with ethyl acetate. The aqueous layer was saturated with ammonium chloride and then extracted with tetrahydrofuran. The extract was washed with aqueous sodium chloride solution and dried over anhydrous magnesium sulfate. The solvent was then removed by distillation under reduced pressure. The residue was purified by column chromatography on silica gel eluting with a mixture of ethyl acetate-methanol (5: 1 by volume) to give 2.34 g.

II

92487 63 g of the title compound as colorless crystals with a melting point of 53-54 ° C.

Nuclear Magnetic Resonance Spectrum (CDCl 3, 60 MHz) δ ppm: δ 1.42-3.89 (6H, multiplet); 2.74 (3H, doublet, J = 5.0 Hz); 3.66 (2H, singlet); 3.76 (3H, singlet); 6.79, 7.17 (4H, A2B2, J = 9.0 Hz); Δ 7.03-7.75 (2H, multiplet).

3- (S) (2S, 4S) -4 - (, 4-methoxybenzylthio) -1-methyl-2-methylcarbamoylpyrrolidine 15,244 μΐ of methyl iodide and 300 mg of sodium bicarbonate were added to an ice-cooled solution containing 1.00 g. (2S, 4S) -4- (4-methoxy-benzylthio) -2-methylcarbamoylpyrrolidine [prepared as described in step (4) above] dissolved in 6 ml of dry dimethylformamide. The mixture was then stirred at 0-5 ° C for 20 hours and allowed to stand overnight at room temperature. At the end of this period, the reaction mixture was poured into aqueous sodium chloride solution and extracted with ethyl acetate. The extract was washed with aqueous sodium chloride solution and dried over anhydrous magnesium sulfate. The solvent was then removed by distillation under reduced pressure and ice-dried. 25 nos were purified by column chromatography on silica gel eluting with a mixture of ethyl acetate and methanol (10: 1 by volume) to give 222 mg of the title compound as colorless crystals, m.p. 82-84 ° C.

30 Infrared absorption spectrum (KBr) vmax cm'1:; 1655, 1512, 1251.

64 92487

Nuclear Magnetic Resonance Spectrum (CDCl 3, 60 MHz) δppm: 1.45-3.91 (6H, multiplet); 2.31 (3H, singlet); 2.80 (3H, doublet, J = 5.0 Hz); Δ 3.66 (2H, singlet); 3.77 (3H, singlet); 6.81, 7.20 (4H, A2B2, J = 9.0 Hz); 6.85-7.60 (1H, multiplet).

10 3 - (6) (2S. 4S) -1. 1-Dimethyl-4-methylcarbamoyl-4- (4-methoxybenzylflithio'-pyrrolidinium fluorosulfate 280 μl of methyl fluorosulfonate was added under ice-cooling to a solution of 210 mg of (2S, 4S) -4-methoxybenzyl-4-methoxybenzyl -15. was washed several times with diethyl ether between decanters and dried under reduced pressure to give 281 mg of the title compound as a colorless powder.

Infrared absorption spectrum (KBr) Pmax cm -1: 25 1681, 1512, 1248.

Nuclear Magnetic Resonance Spectrum (CDCl 3, 270 MHz) δ ppm: 1.95 - 2.43 (2H, multiplet); 2.62 (3H, singlet); Δ 2.61-3.88 (3H, multiplet); : 3.03 (3H, singlet); 3.10 (3H, singlet); 3.65 (3H, singlet); 3.68 (2H, singlet); 35.12 (1H, triplet, J = 8.06 Hz); 6.83, 7.12 (4H, A2B2, J = 8.61 Hz).

II

»„ 92487 οΓ)

Preparation 4 12S, 4S) -1,1-Dimethyl-2- (N, N-dimethylcarbamoyl) -4- (4-methoxybenzylthio) Dvrolidinium fluorosulfonate 5 4 - (1) (2S, 4R) -1- (t- butoxycarbonyl) -2- (N, N-dimethylcarbamoyl-4-hydroxypyrrolidine 3.84 ml of triethylamine was added at -15 to -20 ° C to a solution of 5.8 g of (2S, 4R) -1 - (t-butolscarbonyl) -4-hydroxy-2-pyrrolidinecarboxylic acid dissolved in 85 ml of dry tetrahydrofuran, and then a solution of 2.63 ml of ethyl chloroformate in 25 ml of dry tetrahydrofuran was added to the resulting mixture at this same temperature. The reaction mixture was then stirred for 2 hours, after which 19.75 ml of a 50% v / v aqueous solution of dimethylamine was added at -20 to -25 ° C. The mixture was then stirred for 3 hours under ice-cooling, after which it was allowed to cool. After this time, the mixture was poured into a mixture of 30 ml of concentrated chlorine. -20 hydrochloric acid and ice, and extracted with ethyl acetate. The extract was washed with aqueous sodium chloride solution and dried. The solvent was then removed by distillation under reduced pressure, and the residue was purified by column chromatography on silica gel eluting with a mixture of ethyl acetate and methanol (9: 1 by volume) to give 429 mg: 25 of the title compound.

. 4

Nuclear Magnetic Resonance Spectrum (CDCl 3, 60 MHz) δ ppm: 1.42 (9H, singlet); 1.86 - 2.34 (2H, multiplet); 2.58-2.95 (1H, multiplet); · 2.97 (3H, multiplet); 3.10 (3H, singlet); 3.43 - 3.74 (2H, multiplet); 4.36-5.00 (2H, multiplet).

35 66 92487 4- (2) [2S, 4R) -1- (t-Butoxycarbonyl) -2- (N, N-dimethylcarbamoyl) -methanesulfonylloxy] pyrrolidine 297 μΐ of methanesulfonyl chloride and then 537 μΐ of triethylamine 5 were added and the residue was added in a portion of ice. , 4R) -1- (t-butoxycarbonyl) -2- (Ν, Ν-dimethylcarbamoyl) -4-hydroxypyrrolidine [prepared as described in step (1) above] dissolved in 20 ml of dry tetrahydrofuran. The mixture was stirred at 0-5 ° C for 1 hour and then at room temperature for 10 hours. At the end of this time, the mixture was worked up as described in "Preparation 1- (3>") to give 1.05 g of the title compound as a colorless oil.

Nuclear Magnetic Resonance Spectrum (CDCl 3, 60 MHz) δ ppm: 1.43 (9H, singlet); 2.15 - 2.60 (2H, multiplet); 2.97 (3H, singlet); 3.07 (3H, singlet); 3.10, 3.13 (together 3H, each singlet); 3.83 (2H, doublet, J = 4.0 Hz); 4.63-5.03 (1H, multiplet); 5.15-5.46 (1H, multiplet).

4- (3) (2S, 4S) -1- (t-butoxycarbamoyl) -2- (N, N-dimethylcarbamoyl) -4- (4-methoxybenzyl) pyrrolidine 151 mg sodium hydride (55 w / w as a dispersion) in mineral oil) was added under ice-cooling to a solution of 532 mg of 4-methoxybenzyl mercaptan dissolved in 10 ml of dry dimethyl-30-formoform. The mixture was then stirred at room temperature for 30 minutes, after which 1.05 g of (2R, 4R) -1- (t-butoxycarbonyl) -2- (Ν, Ν-dimethylcarbamoyl) -4-methanesulfonyl-oxypyrrolidine [prepared as described in step (2) above]. The mixture was then stirred at room temperature for 30 minutes and then at 40 ° C for 6 hours. After this period, the reaction mixture

II

92487 67 was treated and purified as described in "Preparation 1- (4)" to give 385 mg of the title compound as an oil.

Infrared absorption spectrum (liquid film) umax cm'1: δ 1690, 1660, 1605, 1585, 1515.

Nuclear Magnetic Resonance Spectrum (CDCl 3, 60 MHz) δ ppm: 1.35 and 1.38 (9H, singlet); 1.55 - 3.37 (5H, multiplet); Δ 2.93 (3H, singlet); 3.00 (3H, singlet); 3.68 (2H, singlet); 3.77 (3H, singlet); 4.30-4.75 (1H, multiplet); 6.85, 7.25 (4H, A2B2, J = 9.0 Hz).

4- (4) (2S, 4S) -2- (N, N-dimethylcarbamoyl) -4- (4-methoxybenzylthio) pyrrolidine 20 ml of a 4N solution of hydrogen chloride in 4N dioxane was added to a cooled solution of 385 mg of (2S, 4S) in ice. 1- (t-butoxycarbonyl) -2- (N, N-dimethylcarbamoyl) -4- (4-methoxybenzylthio) pyrrolidine [prepared as described in step (3) above] dissolved in 1 ml of ethyl acetate, and then the mixture was stirred at room temperature. In 25 patients for 1.5 hours. At the end of this period, it was poured into saturated aqueous sodium bicarbonate solution to weakly acidify it, and then extracted with ethyl acetate. The extract was washed with aqueous sodium chloride solution and dried over anhydrous magnesium sulfate. The solvent was then removed by distillation under reduced pressure. The residue was purified by column chromatography over silica gel. eluting with a mixture of ethyl acetate and methanol (1: 2 by volume) to give 163 mg of the title compound as an oil.

Infrared absorption spectrum (liquid film) umax cm'1: 35 3300, 1640, 1605, 1590, 1510.

»68 92487

Nuclear Magnetic Resonance Spectrum (CDCl 3, 60 MHz) δ ppm: 1.21 - 1.78 (2H, multiplet); 2.09 - 3.98 (5H, multiplet); 2.94 (6H, singlet); Δ 3.64 (2H, singlet); 3.74 (3H, singlet); 6.81, 7.21 (4H, A2B2, J = 9.0 Hz).

4- (S) (2S, 4S) -2- (N, N-dimethylcarbamoyl) -4- (4-methoxybenzylmethyl) -1-methylpyrrolidine 41 μΐ of methyl iodide and 84 mg of sodium bicarbonate were added to an ice-cooled solution containing 163 mg of 2S..4S.) - 2- (N, N-dimethylcarbamoyl) -4- (4-methoxybenzylthio) pyrrolidine [prepared as described in step (4) above] dissolved in 1.5 ml of dry dimethylformamide, and then the mixture was stirred at room temperature for 4 hours. At the end of this period, the reaction mixture was poured into saturated aqueous sodium bicarbonate solution and extracted with ethyl acetate. The extract was washed with aqueous sodium chloride solution and dried over anhydrous magnesium sulfate. The solvent was then removed by distillation under reduced pressure, and the residue was purified by chromatography on a Lobar column (Merck, LiChroprep Si60, size A). Fractions eluted with a mixture of ethyl acetate and methanol (3: 1 by volume) gave 45 mg of the title compound as an oil.

Infrared absorption spectrum (liquid film) umax cm-1: 1640, 1607, 1580, 1510.

Nuclear Magnetic Resonance Spectrum (CDCl 3, 60 MHz) δ ppm: 1.56-2.18 (2H, multiplet); 2.20 - 3.60 (4H, multiplet); 2.34 (3H, singlet); 2.96 (3H, singlet); Δ 3.10 (3H, singlet); 3.70 (2H, singlet); 11,92487 69 3.78 (3H, singlet); 6.82, 7.22 (4H, A2B2, J = 9.0 Hz).

4- (6) Methyl 1- (t-butoxycarbonyloxy) -4-hydroxyprolinate 5 2.2 g of sodium hydride (55% w / w dispersion in mineral oil) were added at 0-5 ° C to a solution containing 1.15 g. g of 1- (t-butoxycarbonyl) -4-hydroxyproline dissolved in 100 ml of dry dimethylformamide. The mixture was then stirred at room temperature for 1.5 hours, after which it was cooled to 0-5 ° C and 3.42 ml of methyl iodide was added. The reaction mixture was then allowed to stand overnight at room temperature. The reaction mixture was then poured into saturated aqueous sodium chloride solution and extracted with ethyl acetate. The extract was washed with aqueous sodium chloride solution and dried, and then the solvent was removed by distillation under reduced pressure. The residue was purified by chromatography on silica gel eluting with a mixture of benzene and ethyl acetate (1: 1 by volume) to give 6.1 g of the title compound as an oil.

20

Infrared absorption spectrum (liquid film) vmax cm -1: 3430, 1750, 1700, 1670.

Nuclear Magnetic Resonance Spectrum (CDCl 3, 60 MHz) δ ppm: 1.41 (9H, singlet); 1.78 - 2.84 (3H, multiplet); 3.58 (2H, doublet, J = 4.0 Hz); 3.71 (3H, singlet); 4.18-4.62 (2H, multiplet).

4- (7) (2S, 4R) -1-tert-Butoxycarbonyl) -4-methanesulfonyl-hydroxy-2-methoxycarbonyl-pyrrolidine 2.02 ml of methanesulfonyl chloride and then 3.65 ml of triethylamine 35 were added to a solution of 6 g cooled in ice. - (t-butoxycarbonyl) -4-hydroxyprolinate [prepared as described in step (70) above (9)] dissolved in 120 ml of dry tetrahydrofuran. The mixture was first stirred at 0-5 ° C for 1 hour and then at room temperature for 1 hour, after which it was poured into aqueous sodium chloride solution and extracted with ethyl acetate. The extract was washed with aqueous sodium chloride solution and dried over anhydrous magnesium sulfate. The solvent was removed by distillation under reduced pressure to give 8.13 g of the title compound as an oil.

Nuclear Magnetic Resonance Spectrum (CDCl 3, 60 MHz) δ ppm: 1.47 (9H, singlet); 1.74 - 2.85 (2H, multiplet); 3.08 (3H, singlet); 3.79 (3H, singlet); 3.81 (2H, doublet, J = 4.0 Hz); 4.15-4.65 (1H, multiplet); 5.12-5.41 (1H, multiplet).

4- (8) (2S, 4S) -1- (t-butoxycarbonyl) -4- (α-methoxybenzyllithio) -2-20 methoxycarbonylpyrrolidine 1.11 g of sodium hydride (55% w / w dispersion in mineral oil) was added in ice. cooling to a solution of 3.51 ml of 4-methoxybenzyl mercaptan dissolved in 60 ml of dry dimethylformamide and then the mixture was stirred at room temperature for 30 minutes. At the end of this time, a solution of 8.13 g of (2S, 4S) -1- (t-butoxycarbonyl) -4-methanesulfonyloxypyrrolidine [prepared as described in step (7) above] in 20 ml of dry dimethylformamide was added dropwise to the mixture. and then the mixture was stirred at room temperature for 15 minutes and then at 40 ° C for 4 hours. The reaction mixture was then poured into aqueous sodium chloride solution and extracted with ethyl acetate. The extract was washed with water, dried, and the solvent was removed by distillation under reduced pressure. The residue was purified by chromatography on silica gel eluting with a mixture of benzene and ethyl acetate (v / v).

II

92487 71: 15: 1) to give 6.89 g of the title compound 61.

Infrared absorption spectrum (liquid film) umax cm -1: 5 1745, 1690, 1605, 1580, 1510.

Yunar magnetic resonance spectrum (CDCl 3, 60 MHz) δ ppm: 1.40 (9H, singlet); 1.71 - 3.48 (5H, multiplet); Δ 3.71 (3H, singlet); 3.78 (2H, singlet); 4.01-4.45 (1H, multiplet); 6.85, 7.25 (4H, A2B2, J = 9.0 Hz).

4- (9) (2S, 4S1) -4- (4-methoxybenzyl litio) -2-methoxycarbonylpyrrolidine 27.3 ml of a 4N ethyl acetate solution of hydrogen chloride were added to a cooled solution of 5.22 g of (2S, 4S) in ice. -1- (t-Butoxycarbon-20-yl) -4- (4-methoxybenzylthio) -2-methoxycarbonylpyrrolidine [prepared as described in step (8) above] dissolved in 14 ml of ethyl acetate, and then the mixture was stirred at room temperature for 1 hour. At the end of this period, it was poured into saturated aqueous sodium bicarbonate solution and extracted with ethyl acetate. The extract was washed with aqueous sodium chloride solution and dried over anhydrous magnesium sulfate. The solvent was then removed by distillation under reduced pressure to give 3.3 g of the title compound as an oil.

Infrared absorption spectrum (liquid film) umax cm -1: 3250, 1735, 1610, 1580, 1510.

92487 72

Nuclear Magnetic Resonance Spectrum (CDCl 3, 60 MHz) δ ppm: 1.56-3.32 (6H, multiplet); 3.67 (2H, singlet); 3.72 (3H, singlet); Δ 3.78 (3H, singlet); 3.70-3.99 (1H, multiplet).

4- (10) (2S, 4S) -4- (4-methoxybenzyl) thio-2-methoxycarbonyl-1-methylpyrrolidine 1.18 g of sodium bicarbonate and 0.876 ml of methyl iodide were added to a cooled solution of 3.3 g (2S, 4S) -4- (4-Methoxybenzylthio) -2-methoxycarbonylpyrrolidine [prepared as described in step (9) above] dissolved in 30 ml of dry dimethylformamide, and then the mixture was stirred at room temperature for 5 hours. At the end of this period, the reaction mixture was poured into saturated aqueous sodium bicarbonate solution and extracted with ethyl acetate. The extract was washed with aqueous sodium chloride solution and dried over anhydrous magnesium sulfate. The solvent was then removed by distillation under reduced pressure. The residue was purified by column chromatography on silica gel eluting with a mixture of ethyl acetate and benzene (1: 2 by volume) to give 968 mg of the title compound as an oil.

Infrared absorption spectrum (liquid film) t> max cm -1: 1740, 1730, 1605, 1580, 1510.

Nuclear Magnetic Resonance Spectrum (CDCl 3, 60 MHz) δ ppm: 1.64 - 3.34 (6H, multiplet; 2.37 (3H, singlet)); 3.70 (2H, singlet); 3.81 (3H, singlet); singlet), 3.88 (3H, singlet), 6.83, 7.23 (4H, A2B2, J = 9.0 Hz).

35 4- (11) (2S, 4S) -2- (N, N-dimethylcarbamoyl) -4- (4-methoxybenzylthio) -1-methylpyrrolidine 73,92487 1.92 ml of 1N aqueous sodium hydroxide solution was added to a solution containing 378 mg. (2S, 4S) -4- (4-methoxybenzylthio) -2-methoxycarbonyl-1-methylpyrrolidine [prepared as described in (10) above] dissolved in 3.84 ml of methanol, and the mixture was stirred at room temperature for 3 hours. At the end of this period, the mixture was neutralized by adding 1.92 ml of 1N aqueous hydrogen chloride. The solvent of the site was removed by distillation under reduced pressure, and the residue was dried to give (2S, 4S) -2-carboxy-4- (4-methoxybenzylthio) -1-methylpyrrolidine as a crude product.

This crude compound was completely suspended in 7.3 ml of aceto-15-nitrile, and 318 mg of N, N'-carbonyldiimidazole was added to this suspension, after which the resulting mixture was stirred at 40 ° C for 1 hour. At the end of this period, the mixture was allowed to cool to room temperature, and then a solution of 559 mg of dimethylamine in 3.7 ml of tetrahydrofuran was added to the mixture. The mixture was allowed to stand at room temperature overnight. The solvent and excess dimethylamine were removed by distillation under reduced pressure. The residue was mixed with aqueous sodium chloride solution and extracted with ethyl acetate. The extract was dried over anhydrous magnesium sulfate, and the solvent was distilled off under reduced pressure. Residue 25 was purified as described in Reference Example 4- (5) to give 382 mg of the title compound as an oil.

4- (12) (2S, 4S) -2- (N, N-dimethylcarbamoyl) -4- (4-methoxybenzylthio) -1,1-dimethylfluorolidinium fluorosulfonate 30. 139 μΐ of methyl fluorosulfonate was added to an ice-cooled solution containing 190 mg of (2S, 4S) -2- (N, N-dimethylcarbamoyl) -4- (4-methoxybenzylthio) -1-methylpyrrolidine [prepared in step (5) above or (11 ) dissolved in 3.8 ml of methylene chloride, and this mixture was stirred at room temperature 92487 74 for 2 hours. At the end of this period, the solvent was removed by distillation under reduced pressure. The residue was washed several times with hexane between decanters and dried under reduced pressure to give 356 mg of the title compound as an oil.

5

Nuclear Magnetic Resonance Spectrum (D 2 O, 60 MHz) δ ppm: 1.58-4.58 (12H, multiplet); 3.07 (6H, singlet); 3.84 (3H, singlet); Δ 3.90 (2H, singlet); 6.90, 7.25 (4H, A2B2, J = 9.0 Hz).

Preparation 5 (3S) -1,1-Dimethyl-3- (4-methoxybenzyl) pyrrolidinurofluorosulfonate 5- (1R (3R) -1 · -t-butoxycarbonyl-3-methanesulfonyl] pyrrolidinidine 16.91 ml of triethylamine and 9.36 ml of methanesulfonyl chloride were added in this order to an ice-cooled solution containing 25 g of (3R) -lt-butoxycarbonyl-3-hydroxypyrrolidine and then dissolved in 250 ml of tetrahydrofuran, dried in 250 ml of urea. The mixture was poured into aqueous sodium chloride solution and extracted with ethyl acetate, and the extract was washed with aqueous sodium chloride solution and dried over anhydrous magnesium sulfate. by distillation under reduced pressure to give 3.10 g of the title compound as a colorless oil.

75 92487

Nuclear Magnetic Resonance Spectrum (CDCl 3, 60 MHz) δ ppm: 1.48 (9H, singlet); 1.91 - 2.45 (2H, multiplet); 3.04 (3H, singlet); Δ 3.26-3.82 (4H, multiplet); 6.1-6.44 (1H, multiplet).

5- (2) OS ') - tert-Butoxycarbonyl-3- (4-methoxybenzyl) pyrrolidine 5.32 g of sodium hydride (55% w / w dispersion in mineral oil) was added under ice-cooling to a solution containing 316 g / l. 86 ml of 4-methoxybenzyl mercaptan dissolved in 200 ml of dry dimethylformamide and then the mixture was stirred at room temperature for 15 minutes in a patient. At the end of this time, a solution of 31.0 g of (3R) -1-t-butoxycarbonyl-3-methanesulfonyloxypyrrolidine [prepared as described in step (1) above] in 50 ml of dry dimethylformamide was added to the mixture. The mixture was then stirred for 30 minutes under ice-cooling, after which it was allowed to stand overnight at room temperature. The reaction mixture was then poured into aqueous sodium chloride solution and extracted with ethyl acetate. The extract was washed with aqueous sodium chloride solution and dried over anhydrous magnesium sulfate. The solvent was removed by distillation under reduced pressure. The residue was purified by chromatography on silica gel eluting with a mixture of hexane and ethyl acetate (5: 1 by volume) to give 28.0 g of the title compound as a light brown oil.

Nuclear Magnetic Resonance Spectrum (CDCl 3, 60 MHz) δ ppm: 1.46 (9H, singlet); . 1.50 - 2.35 (2H, multiplet); 2.81 - 3.88 (5H, multiplet); 3.70 (2H, singlet); 3.79 (3H, singlet); Δ 6.83, 7.27 (4H, A2BZ, J = 9.0 Hz.

5- (3) (3S) -3- (4-Methoxybenzyl) pyrrolidine hydrochloride 76,92487 106 ml of a 4N ethyl acetate solution of hydrogen chloride were added to a cooled solution of 27.50 g of (3R) -1-t-butoxycarbonyl. 3- (4-methoxybenzylthio) pyrrolidine [prepared as described in step (2) above] dissolved in 100 ml of ethyl acetate, and then the mixture was stirred at 0-5 ° C for 30 minutes and then at 25 ° C for 2 hours. . At the end of this period, the mixture was diluted with 200 ml of diisopropyl ether, and the precipitated crystals were collected by filtration to give 20.84 g of the title compound as colorless crystals, m.p. 125-126 ° C.

Infrared absorption spectrum (KBr) umax cm'1: 1510, 1246, 1174.

15

Nuclear Magnetic Resonance Spectrum (CDCl 3, 60 MHz) δ ppm: 1.52-2.53 (2H, multiplet); 2.91 - 3.70 (5H, multiplet); 3.63 (2H, singlet); 3.67 (3H, singlet); 6.80, 7.16 (4H, A2B2, J = 9.0 Hz).

5- (4) (3S) -3- (4-methoxybenzylmethyl) -1-methylpyrrolidine. 750 mg of (3S) -3- (4-methoxybenzylthio) pyrrolidine [prepared by neutralizing 900 mg of (3S) -3- (4-methoxybenzylthio) pyrrolidine hydrochloride prepared as described in (3) above with sodium bicarbonate] was dissolved in 15 mg. To 1 ml of dry acetonitrile and to this solution was added 1.44 ml of a 35% v / v aqueous solution of formaldehyde.

The reaction mixture was then stirred for 15 minutes, after which it was neutralized by the addition of acetic acid; it was further stirred for 2.5 hours. At the end of this period, the reaction mixture was poured into 200 ml of ethyl acetate, and the mixture was washed with 2N aqueous potassium hydroxide solution and aqueous sodium chloride solution. The organic layer was dried over potassium carbonate, and the solvent was removed by distillation under reduced pressure. The residue was purified by column chromatography over silica gel.

II

92487 77 eluted with a mixture of ethyl acetate and methanol (3: 1 by volume) to give 349 mg of the title compound as a colorless oil.

Nuclear Magnetic Resonance Spectrum (CDCl 3, 60 MHz) δ ppm: 1.40 - 3.49 (7H, multiplet); 2.33 (3H, singlet); 3.69 (2H, singlet); 3.78 (3H, singlet); Δ 6.86, 7.25 (4H, A2B2, J = 9.0 Hz).

5- (5) (3S) -1,1-Dimethyl-3- (4-methoxybenzyl) pyrrolidinium fluorosulfonate 118 μΐ of methyl fluorosulfonate was added to an ice-cooled solution containing 340 mg of (3S) -3- (4-methoxybenzyl). 1-methylpyrrrolidine [prepared as described in step (4) above] dissolved in 20 ml of methylene chloride. The mixture was then stirred at this same temperature for 30 minutes and then at room temperature for 20 hours. At the end of this period, the solvent was removed by distillation under reduced pressure, and the residue was washed several times with diethyl ether between decanters and dried under reduced pressure to give 500 mg of the title compound as a colorless powder.

. 25

Nuclear Magnetic Resonance Spectrum (D 2 O, 270 MHz) δ ppm: 1.86-2.05 (1H, multiplet); 2.34-2.56 (1H, multiplet); 2.90 (3H, singlet); 3.01 (3H, singlet); · 2.98-3.73 (5H, multiplet); 3.65 (3H, singlet); 3.67 (2H, singlet); 6.82, 7.17 (4H, A2B2, J = 8.62 Hz).

35 1,1-Dimethyl-4- (4-methoxybenzyl) piperidinium fluorosulfonate 78,92487

Preparation 6 6- (1) 4-Methanesulfonyloxy-1-methylpiperidine 13.3 ml of triethylamine and then 7.4 ml of methanesulfonyl chloride were added to a cooled solution of 10 g of 4-hydroxy-1-methylpiperidine dissolved in 100 ml. dry tetrahydrofuran-10. The mixture was then stirred at 0-5 ° C for 2 hours and then at room temperature for 1.5 hours, after which it was poured into aqueous sodium chloride solution and extracted with ethyl acetate. The extract was washed with aqueous sodium chloride solution and dried over anhydrous magnesium sulfate. The solvent was then removed by distillation under reduced pressure to give 12.74 g of the title compound as an oil.

Nuclear Magnetic Resonance Spectrum (CDCl 3, 60 MHz) δ ppm: 1.74 - 2.90 (8H, multiplet); 2.28 (3H, singlet); 3.00 (3H, singlet); 4.73 (1H, multiplet).

6- (2) 4- (4-methoxybenzylthio) -1-methylpiperidine. 10.9 ml of 4-methoxybenzyl mercaptan were dissolved in 55 ml of dry dimethylformamide, and 3.4 g of sodium hydride (55% w / w suspension in mineral oil) was added to this solution under cooling. The mixture was then stirred at room temperature for 30 minutes.

At the end of this period, a solution containing: 12.6 mg of 4-methanesulfonyloxy-1-methylpiperidine in 63 ml of dimethylformamide was added to the mixture, and then it was allowed to stand overnight at room temperature. The reaction mixture was then poured into aqueous sodium chloride solution and extracted with ethyl acetate. The extract was washed with aqueous sodium chloride solution and dried over anhydrous magnesium sulfate. The solvent was then removed by distillation under reduced pressure

II

92487 79 and the residue was purified by chromatography on silica gel eluting with a mixture of ethyl acetate and methanol (1: 4 by volume) to give 9.01 g of the title compound as an oil.

Nuclear Magnetic Resonance Spectrum (CDCl 3, 60 MHz) δ ppm: 1.40-3.05 (9H, multiplet); 2.23 (3H, singlet); 3.69 (2H, singlet); 3.78 (3H, singlet); Δ 6.82, 7.23 (4H, A2B2, J = 9.0 Hz).

6- (3) 1,1-Dimethyl-4- (4-methoxybenzyl) piperidinium fluorosulfonate 2.9 ml of methyl fluorosulfonate was added to an ice-cooled solution containing 8.95 g of 4- (4-methoxybenzylthio) -1- methylpiperidine dissolved in 300 ml of dry methylene chloride, and then the mixture was stirred at this same temperature for 20 minutes and then at room temperature for 2.5 hours. At the end of this period, the solvent was removed by distillation under reduced pressure, and the residue was washed several times with diethyl ether between decanters and dried under reduced pressure to give 12.76 g of the title compound as a colorless powder.

; Nuclear Magnetic Resonance Spectrum (CDCl 3, 270 MHz) δ ppm: 1.75 - 1.93 (2H, multiplet); 2.02 - 2.15 (2H, multiplet); 2.70 - 2.84 (2H, multiplet); 3.05 (3H, singlet); 3.06 (3H, singlet); • 3.24-3.97 (4H, multiplet); 3.74 (3H, singlet); 3.78 (2H, singlet); 6.88, 7.26 (4H, A2B2, J = 8.79 Hz).

35 80 92487

Preparation 7 (2S, 4S) -2- (N, N-dimethylcarbamoyl) -1-ethyl-1-methyl-4- (N-methoxybenzyl) pyrrolidinium fluorosulfonate 5- (1-) (2S, 4S) - 1-Ethyl 4- (4-methoxybenzyl) -2-methoxycarbonylpyrrolidinium fluorosulfonate 1.2 g of (2S, 4S) -1-ethyl-4- (4-methoxybenzylthio) -2-methoxycarbonyl-10-pyrrolidine [prepared in "Preparation 4- (9)"] was dissolved in 12 ml of dry dimethylformamide, and 358 mg of sodium bicarbonate and 0.4 ml of ethyl iodide were added to the resulting solution under ice-cooling. The mixture was then stirred at room temperature for 6 hours and then at 45-50 ° C for 3 hours-15 hours. It was then poured into saturated aqueous sodium bicarbonate solution and extracted with ethyl acetate. The extract was washed with aqueous sodium chloride solution and dried over anhydrous magnesium sulfate.

The solvent was then removed by distillation under reduced pressure, and the residue was purified by column chromatography on silica gel eluting with a mixture of ethyl acetate and benzene (1: 4 by volume) to give 904 mg of the title compound as an oil.

Infrared absorption spectrum (liquid film) umax cm -1: 1740, 1615, 1590, 1515.

: 25

Nuclear Magnetic Resonance Spectrum (CDCl 3, 60 MHz) δ ppm: 1.03 (3H, triplet, J <7.0 Hz); 2.72 - 3.43 (8H, multiplet); 3.70 (5H, singlet); 3.78 (3H, singlet); : 6.84, 7.25 (4H, A2B2, J = 9.0 Hz).

Il 92487 81 7- (2) (2S, 4S) -2- (N, N-dimethylcarbamoyl) -1-ethyl-4- (4-methoxybenzylthio) pyrrolidine 883 mg (2S, 4]> ) -1-Ethyl-4- (4-methoxybenzylthio) -2-methoxycarbon-5-ylpyrrolidine was dissolved in 8.6 ml of methanol, and 4.3 ml of 1N aqueous sodium hydroxide solution was added to the solution thus obtained. The mixture was then stirred at room temperature for 2 hours, after which it was neutralized by adding 4.3 ml of 1N aqueous hydrochloric acid. The reaction mixture was evaporated to dryness under reduced pressure to give a crude product.

This crude compound was suspended in 18 ml of dry acetonitrile, and 694 mg of N, N'-carbodiimidazole was added to this suspension. The resulting mixture was then stirred at 40 ° C for 1 hour, after which a solution of 1.89 ml of dimethylamine in 10 ml of tetrahydrofuran was added to the mixture. The mixture was allowed to stand at room temperature overnight, after which it was concentrated by evaporation under reduced pressure. The residue was mixed with aqueous sodium chloride solution and extracted with ethyl acetate, and then dried over aqueous magnesium sulfate. The solvent was distilled off under reduced pressure, and the residue was subjected to column chromatography using a Lobar column (Merck, LiChroprep Si60, size B). The title compound was obtained as an oil from fractions eluted with a mixture of ethyl acetate and methanol (10: 1 by volume).

25

Infrared absorption spectrum (liquid film) umax cm’1: 1640, 1610, 1585, 1515.

Nuclear Magnetic Resonance Spectrum (CDCl 3, 60 MHz) δ ppm: 30 1.01 (3H, triplet, J = 7.0 Hz); : 1.57-3.73 (8H, multiplet); 2.91 (3H, singlet); 3.16 (3H, singlet); 3.70 (2H, singlet); 35 3.78 (3H, singlet); 6.83, 7.27 (4H, A2B2, J = 9.0 Hz).

92487 82 7- (3) (2S, 4S) -2- (N, N-dimethylcarbamoyl) -1-ethyl-1-methyl-4- (4-methoxybenzyl) pyrrolidinium fluorosulfonate 128 μΐ methyl fluorosulfonate was added to a solution containing 5,438 mg (2S, 4S) -2- (Ν, Ν-dimethylcarbamoyl) -1-ethyl-4- (4-methoxybenzylthio) pyrrolidine dissolved in 10 ml of methylene chloride, and then this mixture was stirred at room temperature for 2 hours.

At the end of this period, the solvent was removed by distillation under reduced pressure, and the residue was washed several times with diethyl ether between decanters and dried under reduced pressure to give the title compound as an oil.

Nuclear Magnetic Resonance Spectrum (D 2 O, 270 MHz) δ ppm: 1.12 (3H, triplet, J = 7.15 Hz); 1.96-2.08 (1H, multiplet); 2.62 - 3.51 (6H, multiplet); 2.79 (3H, singlet); 2.93 (3H, singlet); 2.95 (3H, singlet); 3.65 (3H, singlet); 3.68 (2H, singlet); 4.55-4.60 (1H, multiplet).

Preparation 8: 25 (25.45) -1,1-Dimethyl-4- (4-methoxybenzyl) -2- (4-morpholinocarbonyl) pyrrolidinium fluorosulfonate 8- (1) (2S, 4S) -4- ( 4-Methoxybenzyl] -2-methoxycarbonyl-1-methylpyrrolidine 3.43 ml of 35% formalin was added to a solution of 2.25 g of (25.45) -4- (4-methoxybenzylthio) -2-methoxycarbonylpyrrolidine dissolved in 42 ml: in acetonitrile. Thereafter, 804 mg of sodium cyanoborohydride was divided into three portions and added to this mixture over 5 minutes. The mixture was then stirred at room temperature 92487 83 for 30 minutes. At the end of this period, the mixture was cooled, 1.3 ml of acetic acid was added, and the reaction mixture was stirred at room temperature for 40 minutes. Then, 40 ml of 1N aqueous sodium hydroxide solution and aqueous sodium chloride solution were added to the reaction mixture, after which it was extracted with ethyl acetate. The extract was washed with aqueous sodium chloride solution and dried over anhydrous magnesium sulfate. The solvent was then removed by distillation under reduced pressure, and the residue was purified as described in "Preparation 4- (10)" to give 1.31 g of the title compound as an oil.

The infrared spectrum, nuclear magnetic resonance spectrum and thin layer chromatogram of this compound were similar to those of the compound prepared in "Preparation 4- (10)".

8- (2) (2S, 4S) -4- (4-methoxybenzylthio) -1-methyl-2- (4-morpholino-carbonyl) pyrrolidine 5.32 ml of 1N aqueous sodium hydroxide solution was added to a solution containing 1.31 g. (2S, 4S) -4- (4-methoxybenzylthio) -2-methoxycarbonyl-1-methylpyrrolidine dissolved in 11 ml of methanol, and the mixture was stirred at room temperature for 2 hours. At the end of this period, the mixture was neutralized with 1.92 ml of 1N aqueous hydrochloric acid. The solvent was then removed by distillation under reduced pressure, and the residue was dried and concentrated by evaporation under reduced pressure to give 1.5 g of (2S, 4S) -2-carboxy-4- (4-methoxybenzylthio) -1-methylpyrrolidine as a crude product.

500 mg of this crude (2S, 4S) -2-carboxy-4- (4-methoxybenzylthio) -1-methylpyrrolidine was suspended in 10 ml of acetonitrile, and 278 mg of N, N'-carbonyldiimidazole was added to this suspension, followed by 30 The mixture was stirred at 40 ° C: 1 hour. At the end of this period, the mixture was allowed to cool to room temperature, after which 187 μΐ of morpholine was added. The reaction mixture was then stirred at room temperature for 2 hours and then allowed to stand overnight. The solvent and excess morpholine were removed by distillation under reduced pressure, and the residue was mixed with aqueous sodium chloride solution and extracted with ethyl acetate. The extract 84,92487 was dried over anhydrous magnesium sulfate, and the solvent was distilled off under reduced pressure. The residue was subjected to column chromatography using a Lobar column (Merck, LiChroprep Si60, size B), and the title compound was obtained as an oil from fractions eluted with a 3: 1 mixture of cyclohexane, ethyl acetate and methanol (v / v).

Infrared absorption spectrum (liquid film) umax cm'1: 1639, 1610, 1511.

10

Nuclear Magnetic Resonance Spectrum (CDCl 3, 60 MHz) δ ppm: 1.50 - 3.33 (6H, multiplet); 2.31 (3H, singlet); 3.64 (8H, broad singlet); 3.70 (2H, singlet); 3.78 (3H, singlet); 6.82, 7.21 (4H, A2B2, J = 9.0 Hz).

8- (3) (2S, 4S) -1,1-dimethyl- (4-roethoxybenzyl) -2- (4-morpholino-carbonyl) -pyrrolidinium fluorosulfonate 106 μl of methyl fluorosulfonate was added to a solution of 392 mg (2S, 4S) -4- (4-Methoxybenzylthio) -1-methyl-2- (4-morpholinocarbonyl) pyrrolidine dissolved in 8 ml of methylene chloride, and: this mixture was stirred at room temperature for 2 hours. At the end of this period, the solvent was removed by distillation under reduced pressure, and the residue was washed several times with diethyl ether between decanters and dried under reduced pressure to give 446 mg of the title compound as an oil.

Nuclear Magnetic Resonance Spectrum (D 2 O, 270 MHz) δ ppm: 1.80 - 2.03 (1H, multiplet); 2.67 - 3.90 (12H, multiplet); 2.98 (3H, singlet); 35 3.07 (3H, singlet); 3.65 (3H, singlet); 92487 85 3.68 (2H, singlet); 4.61 (1H, doublet of doublets, J = 8.06, 6.23 Hz); 6.82, 7.17 (4H, A2B2, J = 8.61 Hz).

5 Preparation 9 (2S. 4S ~) -1. 1-Dimethyl-4- (4-methoxybenzyl) -2- (1-pyrrolidinocarbonyl) pyrrolidinium fluorosulfonate 109- (1) (25,481-4- (4-methoxybenzyl) -1-methyl-2- ( 1-pyrrolidinecarbonyl) pyrrolidine

However, the procedure described in "Preparation 8- (2)" above was repeated using 500 mg of crude (2S, 4S) -2-carboxy-4- (4-methoxybenzyl-15-lithio) -1-methylpyrrolidine [prepared in "Preparation 8 - (2) "as described], 278 mg of N, N'-carbonyldiimidazole and 178 μΐ of pyrrolidine instead of morpholine to give 440 mg of the title compound.

Infrared absorption spectrum (KBr) λmax cm -1: 1640, 1611, 1511, 1444.

Nuclear Magnetic Resonance Spectrum (CDCl 3, 60 MHz) δ ppm: 1.58-2.78 (6H, multiplet); 2.30 (3H, singlet); 2.90-3.80 (8H, multiplet); 3.70 (2H, singlet); 3.78 (3H, singlet); 6.84, 7.24 (4H, A2B2, J = 9.0 Hz).

30 · '9- (2) (2S. 4S ~) -1. 1-Dimethyl-4- (4-methoxybenzyl) -2- (1-pyrrolidinocarbonyl) pyrrolidinium fluorosulfonate

However, the procedure described in "Preparation 8- (3)" was repeated using 418 mg of (2S, 4S) -4- (4-methoxybenzylthio) -1-methyl-2- (1-> 86,92487 pyrrolidinocarbonyl) pyrrolidine and 118 μΐ of methyl fluorosulfonates. to give 472 mg of the title compound as an oil.

Nuclear Magnetic Resonance Spectrum (D 2 O, 270 MHz) δ ppm: δ 1.68-1.85 (4H, multiplet); 1.94-2.10 (1H, multiplet); 2.69 - 3.85 (7H, multiplet); 2.99 (3H, singlet); 3.03 (3H, singlet); Δ 3.65 (3H, singlet); 3.67 (2H, singlet); 4.41 (1H, doublet of doublets, J = 8.06, 6.60 Hz); 6.81, 7.17 (4H, A2B2, J = 8.62 Hz).

Preparation 10 (2S, 4S) -2-Carbamoyl-4- (4-methoxybenzylthio) -1-ethyl-1-methylpyrrolidinium fluorosulfonate 10- (1) (2S, 4S) -2-carbamoyl-1- Ethyl 4- (4-methoxybenzyl) pyrrolidine 0.362 ml of ethyl iodide and 315 mg of sodium bicarbonate were added to an ice-cooled solution containing 1000 mg of (2S, 4S) -2-carba-25-moyl-4- (4-methoxybenzothio) to 1 ml of dry dimethylformamide, and the mixture was stirred at room temperature for 5.5 hours. At the end of this period, the reaction mixture was poured into saturated aqueous sodium bicarbonate solution and extracted with ethyl acetate. The extract was washed with aqueous sodium chloride solution and dried over anhydrous magnesium sulfate, and then the solvent was removed by distillation under reduced pressure. The residue was subjected to column chromatography on silica gel (Katayama Chemicals Co., Silica Gel 60K070). 560 mg of the title compound were obtained as crystals having a melting point of 124-125 ° C from fractions eluted with ethyl acetate.

li 92487 87

Infrared absorption spectrum (KBr) umax cm-1: 1632, 1509.

Yrunomagnetic resonance spectrum (CDCl 3 + D 2 O, 270 MHz) δ ppm: δ 1.08 (3H, triplet, J = 7.33 Hz); 1.86-1.96 (1H, multiplet); 2.38 - 2.76 (4H, multiplet); 2.99 - 3.18 (3H, multiplet); 3.70 (2H, singlet); Δ 3.80 (3H, singlet); 6.84, 7.21 (4H, A2B2, J = 8.79 Hz).

10- (2) (2S, 4S) -2-Carbamoyl-4- (4-methoxybenzyl) -1-ethyl-1-methylpyrrolidinium fluorosulfonate 0.43 ml of methyl fluorosulfonate was added to a solution cooled in ice containing 1.44 ml. g of (2S, 4S) -2-carbamoyl-4- (4-methoxybenzylthio) -1-ethylpyrrolidine dissolved in 30 ml of dry methylene chloride, and then this mixture was stirred at room temperature for 20 hours. At the end of this period, the solvent was removed by distillation under reduced pressure, and the residue was washed several times with diethyl ether between decanters and dried under reduced pressure to give 2.0 g of the title compound as an oil.

25 Infrared absorption spectrum (liquid film) i> max cm -1: 1698, 1610, 1512.

Yrununagnetic resonance spectrum (D 2 O, 270 MHz) δ ppm: 1.12 (3H, triplet, J = 7.33 Hz); Δ 2.06-2.18 (1H, multiplet); • 2.63-3.83 (6H, multiplet); 2.92 (3H, singlet); 3.64 (3H, singlet); 3.65 (2H, singlet); 35 4.02-4.09 (1H, multiplet); 6.82, 7.16 (4H, A2B2, J = 8.42 Hz).

88 92487

Preparation 11 (2S, 4S) -2-ethylcarbamoyl-1-ethyl-1-methyl-4- (4-methoxybenzylthio) pyrrolidinium fluorosulfonate 5 11- (11) (2S, 4S) -1-t-butoxycarbonyl-2- Ethylcarbamoyl-1-ethyl-4-hydroxypyrrolidine 15.25 ml of triethylamine and then a solution of 10.48 ml of 10 ethyl chloroformate in 50 ml of dry tetrahydrofuran was added at -25 ° C to a solution of 23.13 g of 2S, 4S) -lt-Butoxycarbonyl-4-hydroxy-2-pyrrolidinecarboxylic acid dissolved in 350 ml of dry tetrahydrofuran, and the mixture was then stirred at the same temperature for 30 minutes. At the end of this period, a 70% v / v aqueous solution of ethylamine was added to the mixture at -22 ° C, the reaction temperature was allowed to rise gradually, and the reaction was stopped when the temperature rose to 10 ° C. The reaction mixture was then stirred with a small amount of aqueous sodium chloride solution, after which it was extracted three times with ethyl acetate. The combined extracts were washed with aqueous sodium chloride solution and dried over anhydrous magnesium sulfate. The solvent was then removed by distillation under reduced pressure to give 23.60 g of the title compound as a colorless oil.

Nuclear Magnetic Resonance Spectrum (CDCl 3, 60 MHz) δ ppm: 1.10 (3H, triplet, J = 7.5 Hz); 1.44 (9H, singlet); 1.80 - 4.65 (10H, multiplet); 6.64 (1H, broad singlet).

30 * 11- (2) (2S, 4S) -lt-Butoxycarbonyl-2-ethylcarbamoyl-4-methanesulfonyl-pyrrolidine 13.81 ml of triethylamine and then 7.65 ml of methanesulfonyl chloride 35 were added to the ice-cooled solution in ice-cooled solution. (2S, 4S) -1-t-butoxycarbonyl-2-ethylcarbamoyl-4-hydroxypyrrole 92487 89 dine dissolved in 250 ml of dry tetrahydrofuran, and then the mixture was stirred at 0-5 ° C for 30 minutes. The reaction mixture was poured into aqueous sodium chloride solution and extracted with ethyl acetate. The extract was washed with aqueous sodium chloride solution and dried over anhydrous magnesium sulfate. The solvent was then removed by distillation under reduced pressure to give 26.54 g of the title compound as colorless crystals having a melting point of 138-140 ° C.

Infrared absorption spectrum (KBr) umax cm -1 1:10 1675, 1548, 1348.

Nuclear Magnetic Resonance Spectrum (CDCl 3, 60 MHz) δ ppm: 1.12 (3H, triplet, J = 7.5 Hz); 1.48 (9H, singlet); Δ 2.05-4.59 (7H, multiplet); 3.03 (3H, singlet); 5.07-5.43 (1H, multiplet); 6.58 (1H, broad singlet).

11- (3) (2S, 4S) -lt-butoxycarbonyl-2-ethylcarbamoyl-4- (4-methoxybenzyl) ovrolidine 3.57 g of sodium hydride (55% w / w dispersion in mineral oil) was added under ice-cooling. to a solution of 11.31 ml of 4-methoxybenzyl mercaptan dissolved in 150 ml of dry dimethylformamide and then the mixture was stirred at 0-5 ° C for 30 minutes. A solution of 26.0 g of (2S, 4S) -lt-butoxycarbonyl-2-ethylcarbamoyl-4-methanesulfonyloxypyrrolidine in 100 ml of dry dimethylformamide was then added to the mixture, and the whole was then stirred at room temperature. t * 2 hours. At the end of this period, the reaction mixture was poured into aqueous sodium chloride solution and extracted with ethyl acetate. The extract was washed with aqueous sodium chloride solution and dried over anhydrous magnesium sulfate. The solvent was then removed by distillation under reduced pressure, and the residue was purified by chromatography on silica gel eluting with a mixture of hexane and ethyl acetate (1: 1 by volume) to give 17.0 g of the title compound as colorless crystals, m.p. -94 ° C.

Infrared Absorption Spectrum (KBr) λmax cm -1: δ 1700, 1655, 1406.

Nuclear Magnetic Resonance Spectrum (CDCl 3, 60 MHz) δ ppm: 1.10 (3H, triplet, J = 7.5 Hz); 1.43 (9H, singlet); Δ 1.85-3.96 (7H, multiplet); 3.69 (2H, singlet); 3.78 (3H, singlet); 4.18 (1H, triplet, J = 7.5 Hz); 6.35 (1H, broad singlet); 6.86, 7.24 (4H, A2B2, J = 9.0 Hz).

11- (4) (2S, 48) -2-Ethylcarbamoyl-4- (4-methoxybenzyl) pyrrolidine 41.19 ml of a 4N ethyl acetate solution of hydrogen chloride were added to a solution containing 13.00 g of (2S, 4S) -lt. butoxycarbonyl-2-ethylcarbamoyl-4- (4-methoxybenzylthio) pyrrolidine dissolved in 100 ml of ethyl acetate, and this mixture was stirred at room temperature for 3 hours. At the end of this period, the reaction mixture was poured into saturated aqueous sodium bicarbonate solution and extracted with ethyl acetate. The aqueous layer was saturated with ammonium chloride and extracted with tetrahydrofuran. The combined extracts were washed with aqueous sodium chloride solution and dried over anhydrous magnesium sulfate, and then the solvent was removed by distillation under reduced pressure. The residue was subjected to column chromatography on silica gel eluting with a mixture of ethyl acetate and methanol (87:13 by volume) to give 8.00 g of the title compound as light brown crystals, m.p. 67-68 ° C (355 ° C). Infrared ) umax cm -1: 1649, 1512, 1240.

li 92487 91

Nuclear Magnetic Resonance Spectrum (CDCl 3, 60 MHz) δ ppm: 1.10 (3H, triplet, J = 7.5 Hz); 1.50-2.20 (1H, multiplet); 2.16 (1H, singlet); Δ 2.23-3.96 (7H, multiplet); 3.69 (2H, singlet); 3.78 (3H, singlet); 6.86, 7.25 (4H, A2B2, J = 9.0 Hz); 7.53 (1H, broad singlet).

11- (5) (2S, 4S) -2-ethylcarbamoyl-1-ethyl-4- (4-methoxybenzylthio) pyrrolidine 301 μΐ of ethyl iodide and 314 mg of sodium bicarbonate were added to a cooled solution containing 1.00 g of ice (1.00 g). 2S, 4S) -2-Ethylcarbamoyl-4- (4-methoxybenzylthio) pyrrolidine dissolved in 8 ml of dry dimethylformamide, and then the mixture was stirred at room temperature for 5 hours. At the end of this period, the reaction mixture was poured into aqueous sodium chloride solution and extracted with ethyl acetate.

The extract was washed with aqueous sodium chloride solution and dried over anhydrous magnesium sulfate, after which the solvent was removed by distillation under reduced pressure. The residue was subjected to column chromatography on silica gel eluting with ethyl acetate to give 982 mg of the title compound as colorless crystals, m.p. 60-61 ° C.

♦

Infrared absorption spectrum (KBr) umax cm'1: 1651, 1512, 1252.

Nuclear Magnetic Resonance Spectrum (CDCl 3, 60 MHz) δ ppm: 1.06 (3H, triplet, J = 7.5 Hz); 1.14 (3H, triplet, J = 7.5 Hz); 1.56-3.90 (10H, multiplet); 3.69 (2H, singlet); 3.80 (3H, singlet); Δ 6.82, 7.20 (4H, A2B2, J = 9.0 Hz); 7.30 (1H, broad singlet).

92487 _____ .. --- τ ~ 92 11- (6) (25,451) -2-ethylcarbanoyl-1-ethyl-1-methyl-4- (4-methoxy-benzylthio) pyrrolidinium fluorosulfonate 233 μΐ of methyl fluorosulfonate was added to the ice-cold solution. -5 to a solution of 910 mg of (2S, 4S) -2-ethylcarbamoyl-1-ethyl-4- (4-methoxybenzylthio) pyrrolidine dissolved in 35 ml of dry methylene chloride, and then the mixture was stirred at room temperature for 3.5 hours. At the end of this period, the solvent was removed by distillation at 10 ° C under reduced pressure and the residue was washed with diethyl ether decantation and dried under reduced pressure to give 1.20 g of the title compound as a light brown oil.

Infrared absorption spectrum (liquid film) umax cm'1: 15 1680, 1513, 1250.

Nuclear Magnetic Resonance Spectrum (D 2 O, 60 MHz) δ ppm: 0.93 (3H, triplet, J = 7.33 Hz); 1.12 (3H, triplet, J = 7.33 Hz); 1.95-2.26 (1H, multiplet); 2.56 - 4.50 (9H, multiplet); 2.88 (3H, singlet); 3.65 (3H, singlet); 3.69 (2H, singlet); Δ 6.83, 7.18 (4H, A2B2, J = 8.80 Hz).

Preparation 12 (2S, 4S) -1-Ethyl-1-methyl-2-methylcarbamoyl-1-ethyl-4- (4-methoxy-benzyl) thyrrolidinium fluorosulfonate 12- (1) (2S, 4S) -1- Ethyl 4- (4-methoxybenzyllithio) -2-methylcarbamoylpyrrolidine 35 0.71 g of ethyl iodide and 742 mg of sodium bicarbonate were added to a solution of 2.25 g of (2S, 4S) -4- (4-methoxybenzylthio) - 2-Methyl 93,92487 lycarbamoylpyrrolidine dissolved in 20 ml of dry dimethylformamide, and then the mixture was stirred at 0-5 ° C for 1 hour and then at room temperature for 5 hours. At the end of this period, the reaction mixture was poured into aqueous sodium chloride solution and extracted with ethyl acetate. The extract was washed with aqueous sodium chloride solution and dried over anhydrous magnesium sulfate, after which the solvent was removed by distillation under reduced pressure. The residue was purified by column chromatography eluting with ethyl acetate to give 1.87 g of the title compound as colorless crystals, m.p. 68-70 ° C.

Infrared Absorption Spectrum (KBr) Omax cm'1: 1657, 1511, 1252.

Nuclear Magnetic Resonance Spectrum (CDCl 3, 60 MHz) δ ppm: 1.02 (3H, triplet, J = 7.0 Hz); 1.58 - 3.91 (8H, multiplet); 2.80 (3H, doublet, J = 5.0 Hz); 3.67 (2H, singlet); 3.78 (3H, singlet); 6.85, 7.21 (AH, A2B2, J = 9.0 Hz); 7.10-7.60 (1H, broad singlet).

12- (2) (2S, 4S) -1-Ethyl-1-methyl-2-methylcarbamoyl-A- (A-methoxy-2-benzylmethyl) pyrrolidinium fluorosulfonate 187 ml of methyl fluorosulfonate was added to 700 ml of ice-cooled solution. mg of (2S, AS) -1-ethyl-N- (A-methoxybenzylthio) -2-methylcarbamoylpyrrolidine dissolved in 30 ml of dry methylene chloride, and then the mixture was stirred at room temperature for 4 hours. At the end of this period, the solvent was removed by distillation under reduced pressure, and the residue was washed several times with diethyl ether between decanters and dried under reduced pressure to give 950 mg of the title compound as a colorless oil.

94 92487

Infrared absorption spectrum (liquid film) umax cm -1: 1683, 1565, 1280.

Nuclear Magnetic Resonance Spectrum (D 2 O, 60 MHz) δ ppm: δ 1.12 (3H, triplet, J = 7.33 Hz); 2.20-2.20 (1H, multiplet); 2.58-4.50 (7H, multiplet); 2.60 (3H, singlet); 2.87 (3H, singlet); Δ 3.65 (3H, singlet); 3.68 (2H, singlet); 6.82, 7.17 (4H, A2B2, J = 8.80 Hz).

Preparation 13 15 (2S, 4S) -1,1-Dimethyl-2-ethylcarbamoyl-4- (4-methoxybenzyl) pyrrolidinium fluorosulfonate 13- (1) (2S, 4S) -2-ethylcarbamoyl-4-C4-methoxybenzyl ) -1-20 methylpyrrolidine 3.43 ml of 35% formalin and 804 mg of sodium cyanoborohydride were added to a solution of 2.36 g of (2S, 4S) -2-ethylcarbamoyl-4- (4-methoxybenzylthio) pyrrolidine dissolved in 42 ml of: in acetonitrile, and then the reaction mixture was stirred at room temperature for 40 minutes. At the end of this period, 1N aqueous sodium hydroxide solution was added to the reaction mixture, and the mixture was poured into aqueous sodium chloride solution and extracted with ethyl acetate. The extract was washed with aqueous sodium chloride solution and dried over anhydrous magnesium sulfate, and the solvent was removed by distillation under reduced pressure. The residue was purified by column chromatography on silica gel eluting with ethyl acetate to give 1.93 g of the title compound as colorless crystals.

i 35 Infrared absorption spectrum (KBr) uraax cm -1: 1655, 1513, 1252.

11,92487 95

Nuclear Magnetic Resonance Spectrum (CDCl 3, 60 MHz) δ ppm: 1.12 (3H, triplet, J = 7.5 Hz); 1.50-2.20 (1H, multiplet); 2.31 (3H, singlet); Δ 2.36-3.88 (7H, multiplet); 3.67 (2H, singlet); 3.78 (3H, singlet); 6.83, 7.21 (4H, A2B2, J = 9.0 Hz); 7.20 (1H, broad singlet).

13- (2) (2S, 4S) -1,1-dimethyl-2-ethylcarbamoyl-4- (4-methoxybenzyl) pyrrolidinium fluorosulfonate 264 μ4 of methyl fluorosulfonate was added to ice-cooled Liu-15 containing 7 mg of 2S, 4S) -2-Ethylcarbamoyl-4- (4-methoxybenzylthio) -1-methylpyrrolidine dissolved in 40 ml of dry methylene chloride, and then the mixture was stirred at room temperature for 2 hours. At the end of this period, the solvent was removed by distillation under reduced pressure, and the residue was washed several times with diethyl ether between decanters and dried under reduced pressure to give 1.332 g of the title compound as a colorless oil.

Infrared absorption spectrum (CHCl 3) umax cm-1: • j 25 1680, 1510, 1240.

Nuclear Magnetic Resonance Spectrum (D 2 O, 270 MHz) & ppm: 0.94 (3H, triplet, J = 7.33 Hz); 1.95-2.20 (1H, multiplet); 2.60-2.98 (1H, multiplet); 2.96 (3H, singlet); * 3.00 (3H, singlet); 3.02 - 3.70 (5H, multiplet); 3.64 (3H, singlet); 3.67 (2H, singlet); 3.97 (1H, triplet, J = 7.88 Hz); 6.81, 7.01 (4H, A2B2, J = 8.80 Hz).

»96 92487

Preparation 14 (2S) -1. 1-Dimethyl-4- (4-methoxybenzylthiomethyl) pyrrolidinium fluorosulfonate 5 14- (1) (2S) -1-t-butoxycarbonyl-2- (4-methoxybenzylthiomethyl) -pyrrolidine

However, the procedure described in Example 5- (2) was repeated using 12.16 g of (2S) -1-butoxycarbonyl-2-methanesulfonyloxymethylpyrrolidine [prepared by methanesulfonylation of (2S) -1-t-butoxycarbonyl-2-hydroxymethylpyrrolidine], 7 , 3 ml of 4-methoxybenzyl mercaptan and 2.3 g of sodium hydride (55 w / w% suspension in mineral oil) in 100 ml of dry dimethylformamide gave 13.93 g of the title compound as an oil.

Infrared absorption spectrum (liquid film) umax cm'1: 1690, 1615, 1590, 1518.

Nuclear Magnetic Resonance Spectrum (CDCl 3, 60 MHz) δ ppm: 1.20 - 4.15 (9H, multiplet); 1.45 (9H, singlet); 3.70 (2H, singlet); 3.78 (3H, singlet); Δ 6.82, 7.25 (4H, A2B2, J = 9.0 Hz).

14- (2) (2S) -2- (4-methoxybenzylmethyl) pyrrolidine

However, the procedure described in Example 5- (3) was repeated using 5 g of (2S) -lt-butoxycarbonyl-2- (4-methoxybenzylthiomethyl) pyrrolidine to give 2.51 g of the title compound as colorless crystals having a melting point of 124-126.5 ° C.

Infrared absorption spectrum (trademark Nujol) cm-1: 35 1615, 1585, 1520.

97 92487

Nuclear Magnetic Resonance Spectrum (CDCl 3, 60 MHz) δ ppm: 1.45-2.20 (4H, multiplet); 2.25 - 3.96 (6H, multiplet); 3.76 (5H, singlet); Δ 6.85 & 7.32 (4H, A2B2, J = 9.0 Hz).

14- (3) (2S) -2- (4-methoxybenzylthiomethyl) -1-methylpyrrolidine

However, the procedure described in Example 5- (4) was repeated using 10 g of (2S) -2- (4-methoxybenzylthiomethyl) pyrrolidine, 35% formalin and 848 mg of sodium cyanoborohydride in the presence of 44 ml of acetonitrile to give 883 mg. as the title compound.

15

Infrared absorption spectrum (liquid film) umax cm’1: 1610, 1585, 1510.

Nuclear Magnetic Resonance Spectrum (CDCl 3, 60 MHz) δ ppm: δ 1.40-3.22 (9H, multiplet); 2.26 (3H, singlet); 3.68 (2H, singlet); 3.78 (3H, singlet); 6.83, 7.25 (4H, A2B2, J = 9.0 Hz).

25 14- (4) (2S) -1. 1-Dimethyl-2- (4-methoxybenzylflithiomethyl) Dvrolidinium fluorosulfonate

However, the procedure described in Example 5- (5) was repeated using 30,740 mg of (2S) -2- (4-methoxybenzylthiomethyl) -1-methylpyrrolidine and 243 μΐ of methyl fluorosulfonate in the presence of 22 ml of methylene chloride to give 1.0 g of the title compound. crystals having a melting point of 150-153 ° C.

35 Infrared absorption spectrum (Nujol) umax cm’1: 1610, 1585, 1518.

98 92487

Single nanomagnetic resonance spectrum (D 2 O, 270 MHz) δ ppm: 1.60 - 2.02 (3H, multiplet); 2.26 - 3.51 (6H, multiplet); 2.64 (3H, singlet); Δ 2.85 (3H, singlet); 3.64 (5H, singlet); 6.82, 7.19 (4H, A2B2, J = 8.79 Hz).

Preparation 15 10 (2R) -1. 1-Dimethyl-4- ('4-methoxybenzylthiomethyl') pyrrolidinium fluorosulfonate 15- (1) The methods 15 described in Examples 14- (1), (2), (3) and (4) were repeated, but using as starting material (2R) - 1-t-butoxycarbonyl-2-hydroxymethylpyrrolidine to give the title compound as an oil.

Infrared absorption spectrum (liquid film) uraax cm'1: 20 1610, 1585, 1512.

Nuclear Magnetic Resonance Spectrum (D 2 O, 270 MHz) δ ppm: 1.64 - 2.00 (3H, multiplet); 2.25 - 3.48 (6H, multiplet); • 2.64 (3H, singlet); 2.85 (3H, singlet); 3.64 (5H, singlet); 6.28, 7.18 (4H, A2B2, J = 8.79 Hz).

30

II

92487 99

Preparation 16 (2S, AS ') - 2-Carbamoyl-1- (2-hydroxyethyl) -4- (4-methoxybenzyl) -1-methyl-pyrrolidinium fluorosulfonate 5 16- (1) (2S, 4S) -2 -Carbamoyl-1- (2-hydroxyethyl) -4- (4-methoxybenzyllitio) Dvrolidine 0.175 ml of 2-iodoethanol and 0.16 g of sodium bicarbonate were added to a 10 ° C cooled solution containing 0.5 g of (2S, 4S) - 2-carbamoyl-4- (4-methoxybenzylthio) pyrrolidine [prepared as described in Example 1- (5)] dissolved in 5 ml of dry dimethylformamide, and then this mixture was stirred at this same temperature for one hour, at room temperature 2, 5 hours and at 40 ° C for 15 to 19 hours. The reaction mixture was then poured into saturated aqueous sodium bicarbonate solution and extracted with ethyl acetate. The extract was washed with aqueous sodium chloride solution and dried over anhydrous magnesium sulfate. The solvent was then removed by distillation under reduced pressure, and the residue was subjected to silica gel column chromatography (Wako Pure Chemicals, Ltd .; Wako Gel C-100). 0.465 g of the title compound was obtained as crystals from fractions eluted with a mixture of ethyl acetate and methanol (95: 5 by volume).

Infrared absorption spectrum (KBr) umax cm'1: • 25 1625, 1510, 1243.

Nuclear Magnetic Resonance Spectrum (CDCl 3, 270 MHz) δ ppm: 1.69 (1H, broad singlet); 1.92-1.99 (1H, multiplet); Δ 2.55-2.72 (3H, multiplet); 2.82-2.92 (1H, multiplet); 3.12 - 3.18 (3H, multiplet); 3.67 (2H, triplet, J = 4.03 Hz); 3.71 (2H, singlet); 35 3.80 (3H, singlet); 5.42 (1H, broad singlet); 92487 100 6.85, 7.21 (4H, A2B2, J = 8.8 Hz); 7.35 (1H, broad singlet).

16- (2) (2S, 4S) -2-Carbamoyl-1- (2-hydroxyethyl) -4- (4-methoxybenzyl) -1-methylpyrrolidinium fluorosulfonate 0.108 ml of methyl fluorosulfonate was added dropwise to the solution at room temperature. 0.38 g of (2S, 4S) -2-carbamoyl-1- (2-hydroxyethyl) -4- (4-methoxybenzylthio) pyrrolidine dissolved in 7.5 ml of dry methylene chloride, and then the mixture was stirred at the same temperature for 2 hours. At the end of this period, the solvent was removed by distillation under reduced pressure, and the residue was washed with diethyl ether by decantation and dried under reduced pressure to give 0.396 g of the title compound as an oil.

15

Preparation 17 3- (4-Methoxybenzylthio) -1-methylquinuclidinium fluorosulfonate 17- (1) 3- (4-Methoxybenzyl) quinuclidine

However, the procedure described in Example 6- (2) was repeated using 3.7 g of 3-methanesulfonyloxyquinuclidine (prepared by methanesulfonylation of 3-quinuclidinol), 3.0 ml of 4-methoxybenzyl mercaptan and 0.942 g of sodium hydride (55 w / w% suspension). in mineral oil) to give 1.74 g of the title compound as an oil.

Infrared absorption spectrum (liquid film) umax cm -1: 1620, 1590, 1510, 1247.

30

Nuclear Magnetic Resonance Spectrum (CDCl 3, 60 MHz) δ ppm: 1.02-2.31 (5H, multiplet); 2.33-3.41 (7J, multiplet); 3.67 (2H, singlet); 35 3.82 (3H, singlet); 6.86, 7.26 (4H, A2B2, J = 9.0 Hz).

92487 101 17- (2) -3- (4-methoxybenzyl) -1-methylquinuclidinium fluorosulfonate

However, the procedure described in Example 6- (3) was repeated using 5,756 mg of 3- (4-methoxybenzylthio) quinuclidine and 271 μΐ of methyl fluorosulfonate in the presence of 5 ml of methylene chloride to give 1.078 g of the title compound as an oil.

Infrared absorption spectrum (liquid film) umax cm'1: 10 1609, 1590, 1512, 1490, 1467.

Nuclear Magnetic Resonance Spectrum (D 2 O, 270 MHz) δ ppm: 1.63-2.16 (5H, multiplet); 2.70 (3H, singlet); 2.76 - 3.77 (7H, multiplet); 3.63 (5H, singlet); 6.80, 7.15 (4H, A2B2, J = 8.61 Hz).

Preparation 18 20 (6S, 8S) -1,4-Dimethyl-8- (4-methoxybenzylthio) -5-oxo-4-aza-1-a-azoniabicyclo [4.3.1] nonane fluorosulfonate 18- (1) (2S, 4S) -1- (2-Hydroxyethyl) -4- (4-methoxybenzyl) -2-: 25: methylcarbamoylpyrrolidine 934 μΐ of 2-iodoethanol and 1.01 g of sodium bicarbonate were added to an ice-cooled solution containing 2.80 g (2S). 4S) -4- (4-methoxybenzylthio) -2-methylcarbamoylpyrrolidine dissolved in 18 to 30 ml of dry dimethylformamide, and then the mixture was stirred at 40 ° C for 24 hours. At the end of this period, the reaction mixture was poured into aqueous sodium chloride solution and extracted with ethyl acetate. The extract was washed with aqueous sodium chloride solution and dried over anhydrous magnesium sulfate. The solvent was then removed by distillation under reduced pressure. and the residue was purified by chromatography on silica gel eluting with a mixture of ethyl acetate and methanol 102,92487 (15: 1 by volume) to give 2.22 g of the title compound as colorless crystals, m.p. 88-89.5 ° C.

Infrared absorption spectrum (KBr) amax cm *: 1637, 1510, 1240.

Nuclear Magnetic Resonance Spectrum (CDCl 3, 270 MHz) δ ppm: 1.66 - 2.05 (2H, multiplet); Δ 2.50-2.91 (4H, multiplet); 2.82 (3H, doublet, J = 4.77 Hz); 3.07 - 3.21 (3H, multiplet); 3.56 - 3.75 (2H, multiplet); 3.69 (2H, singlet); 3.80 (3H, singlet); 6.84, 7.20 (4H, A2B2, J = 8.79 Hz); 7.42 (1H, broad singlet).

18- (2) (2S, 4S) -1- (4-Methanesulfonyloxyethyl) -4- (4-methoxybenzyl] thio) -2-methylcarbamoylpyrrolidine 1.06 ml of triethylamine and then 584 μΐ of methanesulfonyl chloride were added to the ice-cooled solution. , 13 g of (2S, 4S) -1- (2-hydroxyethyl) -4- (4-methoxybenzylthio) -2-methylcarbamate: bamoylpyrrolidine dissolved in 45 ml of dry tetrahydrofuran, and then the mixture was stirred at 0-5 °. At C temperature for 30 minutes. At the end of this period, the precipitated crystals were collected by filtration and washed with tetrahydrofuran. The filtrate and washings were combined and dried over anhydrous magnesium sulfate, and the solvent was removed by distillation under reduced pressure to give 2.50 g of the title compound as an oil.

Infrared absorption spectrum (liquid film) wmax cm -1: 1664, 1510, 1350.

35 »103 92487

Nuclear Magnetic Resonance Spectrum (CDCl 3, 60 MHz) δ ppm: 1.55-4.10 (8H, multiplet); 2.81 (3H, doublet, J = 5.0 Hz); 3.05 (3H, singlet); Δ 3.70 (2H, singlet); 3.80 (3H, singlet); 4.18-4.49 (2H, multiplet); 6.87, 7.26 (4H, A2B2, J = 9.0 Hz); 7.55 (1H, broad singlet).

18 18- (3) (6S, 8S) -8- (4-methoxybenzyl) -4-methyl-5-oxo-1,4-diazabiscyclo Γ 4.3.01nonane 347 mg sodium hydride (55 w / w% suspension in mineral oil) was added to an ice-cooled solution containing 2.64 g of (2S, 4S) -1- (2-methanesulfonyloxyethyl) -4- (4-methoxybenzylthio) -2-methylcarbamoylpyrrolidine dissolved in 30 ml of dimethyl amide, and then the mixture was stirred at 0-5 ° C for 30 minutes and then at 30 ° C for 2 hours. The reaction mixture was then poured into aqueous sodium chloride solution and extracted with ethyl acetate. The extract was washed with aqueous sodium chloride solution, dried over anhydrous magnesium sulfate, and the solvent was removed by distillation under reduced pressure. The residue was subjected to column chromatography on silica gel eluting with a mixture of acetonitrile and methanol (25: 1 by volume) to give 1.75 g of the title compound as an oil.

Infrared absorption spectrum (CHCl 3) umax cm -1: 1637, 1508, 1240.

30

Nuclear Magnetic Resonance Spectrum (CDCl 3, 270 MHz) δ ppm: 1.88 - 2.01 (1H, multiplet); 2.59 (1H, doublet of triplets, J = 13.18, 8.06 Hz); 2.75 - 2.98 (3H, multiplet); Δ 2.94 (3H, singlet); 3.00 - 3.24 (3H, multiplet); 92487 104 3.30 (1H, triplet, J = 8.06 Hz); 3.48-3.62 (1H, multiplet); 3.70 (2H, singlet); 3.80 (3H, singlet); Δ 6.84, 7.33 (4H, A2B2, J = 8.80 Hz).

18- (4) (6S, 8S) -1,4-Dimethyl-8- (4-methoxybenzylthio) -5-oxo-4-aza-1-astoniabiscyclo [4.3.0] nonane fluorosulfonate 10,461 μΐ methyl fluorosulfonate was added to the ice-cooled solution. containing 1.71 g of (6S, 8S) -8- (4-methoxybenzylthio) -4-methyl-5-oxo-1,4-diazabicyclo [4.3.0] nonane dissolved in 60 μl of methylene chloride and the mixture was stirred at room temperature for 2 hours. At the end of this period, the mixture was diluted with diethyl ether and the resulting crystals were collected by filtration and dried under reduced pressure to give 2.09 g of the title compound as colorless crystals, m.p. 208-210 ° C.

Infrared absorption spectrum (KBr) umax cm -1: 1651, 1512, 1295.

Nuclear Magnetic Resonance Spectrum (hexadeuterated dimethyl sulphoxide, 270 MHz) δ ppm: • 2.16-2.33 (1H, multiplet); 2.79-2.95 (1H, multiplet); 2.89 (3H, singlet); 3.23 (3H, singlet); 3.38 (1H, broad singlet); Δ 3.56-4.02 (6H, multiplet); * 3.74 (3H, singlet); 3.80 (2H, singlet); 4.37 (1H, triplet, J = 8.30 Hz); 6.90, 7.26 (4H, A2B2, J = 8.79 Hz).

li 35 105

Preparation 19 92487 3-Mercapto-2-pyrrolidinone trifluoromethanesulfonate 5 20 ml of trifluoroacetic acid and 0.41 ml of trifluoromethanesulfonic acid were added to a cooled solution of 949 mg of 3- (4-methoxybenzylthio) in 4 ml of pyrrolidine-2 then this mixture was stirred at room temperature for 30 minutes. At the end of this period, the reaction mixture was concentrated by evaporation under reduced pressure to give a residue which was purified by column chromatography on silica gel eluting with a mixture of ethyl acetate and methanol (10: 1 by volume) to give 728 mg of the title compound.

Nuclear Magnetic Resonance Spectrum (60 MHz, CDCl 3) δ ppm: 1.75-3.05 (3H, multiplet); 3.30 - 04.04 (3H, multiplet); 8.12 (1H, broad singlet); 9.32 (1H, broad singlet).

20

Preparation 20 4-Mercapto-2-pyrrolidinone 20- (1) 15.6 g of triphenylphosphine was added to a suspension of 3 g of 4-hydroxy-2-pyrrolidinone in 200 ml of tetrahydrofuran, and then the mixture was stirred at room temperature for 5 minutes. , after which it was cooled to -20 ° C. To this solution was added dropwise, at -12 to -20 ° C, a solution of 9.3 ml of diethyl azodicarboxylate in 9 ml of tetrahydrofuran. The mixture was then stirred at 0-5 ° C for 5 minutes, after which it was cooled again to -20 ° C. 4.2 ml of thioacetic acid was then added dropwise to the mixture under cooling at -18 to -20 ° C. The mixture was warmed to 0-5 ° C where it was stirred for 2 hours. The reaction mixture was then concentrated by evaporation under reduced pressure. The residue was purified by first column chromatography on silica gel eluting with a mixture of ethyl acetate and methanol (10: 1 by volume) and then by column chromatography on silica gel eluting with a mixture of acetonitrile and benzene (2: 1 by volume) to give 5 g. 4-Acetylthio-2-pyrrolidinone as colorless crystals.

Nuclear Magnetic Resonance Spectrum (CDCl 3, 90 MHz) δ ppm: 2.00-4.44 (5H, multiplet); 2.31 (3H, singlet); 10.13 (1H, broad singlet).

20- (2) 1.82 The product obtained in step (1) was dissolved in 30 ml of methanol. To this solution, which was cooled in ice, 2.3 ml of a 28% (w / v) methanol solution of sodium methoxide was added dropwise, and then the mixture was stirred for 30 minutes. To the ice-cooled reaction mixture was then added 12 ml of 1N hydrochloric acid, after which the mixture was concentrated by evaporation under reduced pressure to give a powdery residue. The residue was extracted with 50 ml of ethyl acetate, and the extract was dried over anhydrous sodium sulfate. The extract was concentrated by evaporation under reduced pressure to give 1.35 g of the title compound as colorless crystals.

Infrared absorption spectrum (KBr) umax cm -1: 1687, 1681, 1250.

: 25

Nuclear Magnetic Resonance Spectrum (CDCl 3, 270 MHz) δ ppm: 1.95 (1H, doublet, J = 7.0 Hz); 2.30 (1H, doublet of doublets, J = 17.2 & 6.6 Hz); 2.80 (1H, doublet of doublets, J = 17.2 & 7.1 Hz); 3.31 (1H, doublet of doublets, J = 9.9, 5.2 &; 0.8 Hz); 3.59-3.73 (1H, multiplet); 3.80 (1H, doublet of doublets, J = 9.9, 7.3 & 0.7 Hz); 35 6.13 (1H, broad singlet).

II

Preparations 21-25 107 92487

The following compounds were prepared following procedures similar to those described in "Preparations 1-4" above.

5

Preparation 21 (2S, 4S) -1,1-Dimethyl-4- (4-methoxy-benzyl) -2-methoxycarbonyl-pyrrolidinium fluorosulfonate 10

Infrared absorption spectrum (liquid film) umax cm -1: 1747, 1512.

Nuclear Magnetic Resonance Spectrum (D 2 O, 270 MHz) δ ppm: 2.16-2.29 (1H, multiplet); 2.74-2.92 (1H, multiplet); 3.05 (3H, singlet); 3.12 (3H, singlet); 3.38 - 3.73 (3H, multiplet); 3.65 (3H, singlet); 3.66 (3H, singlet); 3.68 (2H, singlet); 4.32 (1H, doublet of doublets, J = 10.99 & 7.69 Hz); 7.16, 7.82 (4H, A2B2, J = 8.61 Hz).

: 25

Preparation 22 (2R, 4S) -2-Carbamoyl-4- (4-methoxy-benzylthio) -1,1-dimethyl-pyrrolidinium fluorosulfonate 30

Infrared absorption spectrum (KBr) umax cm -1: 1698, 1512.

35 92487 108

Nuclear Magnetic Resonance Spectrum (D 2 O, 270 MHz) δ ppm: 2.19 - 2.30 (1H, multiplet); 2.57-2.70 (1H, multiplet); 2.93 (3H, singlet); Δ 3.15 (3H, singlet); 3.30 (1H, doublet of doublets, J = 12.09 & 7.51 Hz); 3.47-3.60 (1H, multiplet); 3.66 (3H, singlet); 3.69 (2H, singlet); 3.77 (1H, doublet of doublets, J = 12.09 & 8.06 Hz); 4.26 (1H, triplet, J = 8.43 Hz); 6.82, 7.18 (4H, A2B2, J = 8.61 Hz).

Preparation 23 15 (2R, 4S) -1,1-Dimethyl-2- (N, N-dimethylcarbamoyl) -4- (4-methoxybenzyl) pyrrolidinium fluorosulfonate

Infrared absorption spectrum (liquid film) umax cm -1: 20 1652, 1511.

Nuclear Magnetic Resonance Spectrum (D 2 O, 270 MHz) δ ppm: 2.17 - 2.29 (1H, multiplet); 2.46-2.58 (1H, multiplet); 2.78 (1H, singlet); 2.94 (3H, singlet); 2.98 (3H, singlet); 3.10 (3H, singlet); 3.26 (1H, doublet of doublets, J = 12.09 & 6.96 Hz); 3.51-3.63 (1H, multiplet); 3.65 (3H, singlet); 3.69 (2H, singlet); 3.84 (1H, doublet of doublets, J = 12.09 & 8.62 Hz); 4.75 (1H doublet of doublets, J = 7.69 & 6.96 Hz); Δ 6.82, 7.17 (4H, A2B2, J = 8.78 Hz).

Preparation 24 109 92487 (2S, AS ') -2-Cyclopropylcarbamoyl. 1-Dimethyl 4- (4-methoxybenzyl) pyrrolidinium fluorosulfonate 5

Infrared absorption spectrum (liquid film) vmax cm’1: 1685, 1532, 1512.

Nuclear Magnetic Resonance Spectrum (Dz0, 270 MHz) δ ppm: 0.26-2.66 (4H, multiplet); 2.00 - 3.65 (6H, multiplet); 2.96 (3H, singlet); 2.98 (3H, singlet); 3.65 (3H, singlet); 3.68 (2H, singlet); 3.92 (1H, triplet, J = 7.86 Hz); 6.81, 3.17 (4H, A2B2, J <8.80 Hz).

Preparation 25 (6S, 8S) -5-Oxo-8- (4-methoxybenzylthio) -1-methyl-4-aza-1-azoniabicyclo [4.3.0] nonane fluorosulfonate: 25 Infrared absorption spectrum (KBr) vmax cm 1: 1680, 1609, 1512, 1246.

Nuclear Magnetic Resonance Spectrum (Dz0, 270 MHz) δ ppm: 2.10-2.24 (1H, multiplet); 2.73-2.90 (1H, multiplet); • 3.65 (3H, singlet); 3.67 (2H, singlet); 3.36-3.85 7H, multiplet); 4.18 (1H, triplet, J = 8.43 Hz); Δ 6.82, 7.17 (4H, AZBZ, J = 8.79 Hz).

Claims (4)

A process for the preparation of 2- (heterocyclylthio) carbapenem derivatives of formula (I) useful as a drug: HO H CH3 \ / I C C-H / \ / \ H3C CH_CH C-S-Ra 10. In II (I) C_N_C O COOR5 [bad: Ra refers to a group of formula 1-8 or 1-9: CH2 - C1) 2 II - CH N - R3 \ / J (1-8): CHO - CII 7 (1-9) - CH N - R '' / CH '£ 115 92487 and bad : R3 represents a wet atom or a C1-CA alkyl group, 5 R 5 is hydrogen atom or carboxy protecting group which is selected from a group containing: alkoxycarbonyloxyalkyl, cycloalkylalkoxycarbonyloxyalkyl, cykloalkyloxikarbonyloxialkylgrupper, 10 cycloalkylcarbonyloxyalkyl, which optionally substituted by a lower alkyl group, aliphatic acyloxymethyl groups, higher aliphatic acyloxyalkyl groups terpenyloxycarbonyloxyalkyl and 5-alkyl substituted (2 -oxo-l, 3-dioxolen-4-yl) alkyl groups; Or a pharmaceutically acceptable salt or ester thereof, characterized in that (a) a compound of formula (IV) is allowed: CH 2 CH 3 CH 3 CH CH-R27 • II II <IV> c_u_c ¢ / 2 COOR28 (wherein R 1 is as defined above, R 27 is a group of the formula -OR 29, wherein: R 29 is a diphenylphosphoryl group; and R 28 is a 4-nitrobenzyl group); React with a compound of formula (Vb): HS- (Ra) (Vb) [bad Ra is defined above, and if necessary any active group is protected], to obtain a compound of formula (Vb) VI): 10 HO H CH3 \ / IC CH / \ / \ H3C CH CH CSR "II II (VI) c-N-c 5. XO COOR28 [wherein R1, R28 is as defined above and Rb refers to said group of formula 1-8 or 1-9, if required, any active groups are protected]; (b) and then, if necessary, removing the protecting groups and / or esterifying and / or salting them: the resulting compound to obtain a compound of formula (I) or a pharmaceutically acceptable salt or ester thereof . 2. A process according to claim 1 for preparing: pivaloyloxymethyl-2- (2-oxo-4-pyrrolidinylthio) -6- (1-hydroxyethyl) -Ι-Ιmethyl-1-carbapen-2-em-3-carboxylate, in particular pivaloyloxymethyl (1R, 5S, 6S) -2- (2-oxo-4-pyrrolidinylthio] -6 - [(IR) -1-hydroxyethyl] -1-methyl-1-carbapen-2-em-3-carboxylate; Or a pharmaceutically acceptable salt or ester thereof, characterized in that the reagents and reaction conditions are selected to obtain the desired compound. The process of claim 1 to prepare: (1-methylcyclohexan-1-yl) carbonyloxymethyl-2- (2-oxo-4-pyrrolidinylthio) -6- (1-hydroxyethyl) -1-methyl-1-carbapenyl 2-em-3-carboxylate, especially (1-methylcyclohexan-1-yl) carbonyloxymethyl- (IR, 5S, 6S) -2- (2-oxo-4-pyrrolidinylthio] * 6 - [(IR) - 1-hydroxyethyl] -1-methyl-1-carbapen-2-em-3-carboxylate; or a pharmaceutically acceptable salt or ester thereof, which may be selected so that the reagents and reaction conditions are selected for to get the desired association. The process of claim 1 to prepare: 1- (cyclopentyloxycarbonyloxy) ethyl 2- (2-oxo-4-pyrrolidinylthio) -6- (1-20 hydroxyethyl) -1-methyl-1-carbapen-2-em. 3-Carboxylate, especially 1- (Cyclopentyloxycarbonyloxy) - (IR5S, 6S) -2- (2-Oxo-4-pyrrolidinylthio] -6 - [(IR) -1-hydroxyethyl] -1-methyl-1-carbapen -2-em-3-carboxylate; or a pharmaceutically acceptable salt or ester thereof, as may be drawn, that the reagents and reaction conditions are selected to obtain the desired compound.