WO1995014692A1

WO1995014692A1 - Cabapenem derivatives and processes for preparing the same

Info

Publication number: WO1995014692A1
Application number: PCT/KR1994/000059
Authority: WO
Inventors: Min Sun Chang; Jong In Lim; Nam Sik Kim; Hee Chan Shin; Gye Won Kim; Ji Yong Kim; Jae Keol Rhee; Chon Woo Lee; Weon Bin Im; Dong Sung Kim
Original assignee: Dong-A Pharmaceutical Co., Ltd.
Priority date: 1993-11-23
Filing date: 1994-05-27
Publication date: 1995-06-01
Also published as: KR950014111A

Abstract

Carbapenem(3-pyrrolidinylthio-1-azabicyclo[3,2,0] hept-2-ene-7-one-carboxylic acid compounds and their derivatives. The compounds of the present invention are useful as antibacterial agents against gram positive, gram negative, imipenem-resistant, clinical isolates of Pseudomonas aeruginosa, and beta-lactamase producing bacteria. Also provided in the present invention are processes for the preparation of the antibacterial compound of the present invention.

Description

CARBAPENEM DERIVATIVES AND PROCESSES

FOR PREPARING THE SAME

Background of the Invention

Field of the Invention

The present invention relates to novel carbapenem

(3-pyrrolidinylthio-1-azabicyclo[3,2,0]hept-2-ene-7-one-2-carboxylic acid compounds) derivatives, processes for preparing the derivatives and antibiotics containing such compounds as active ingredients.

Description of the Related Art

Thienamycin, which is structurlly related to carbapenem, has strong antibacterial activities, however, thienamycin itself is chemically unstable and has been reported to be decomposed by enzymes such as mammalian renal dehydropeptidase I (hereinafter referred to DHP-I), whereby the antibacterial activities decrease. Also the decomposed products are toxic to the kidneys. Thienamycin must be used in combination with a DHP-I inhibitor.

Merck & Co. Inc. have synthesized many thienamycin analogues to maintain the excellent antibacterial activities of thienamycin and to secure chemical stability. As a result, imipenem obtained by formimidation of the amino group of thienamycin, hasbeen developed as a pharmaceutical product (U.S. Pat.

4,194,047). However, like thienamycin, imipenem is likely to be decomposed by DHP-I in the human kidneys.

Therefore, it cannot be used for treatment of infection in the genito-urinary tract. Furthermore the decomposed products are toxic to the kidneys. Therefore, imipenem cannot be administered alone and is required to be used in combination with a DHP-I inhibitor such as cilastatin (Antimicrob. Agents Chemother., vol 12 (Suppl. D), 1(1983)).

However, in the 1980's, there was a growing demand for stable carbapenem compounds, and various carbapenem compounds having a 1β-methyl carbapenem basic skeleton were developed. Sumitomo Co. has synthesized meropenem which is much more stable against DHP-I than imipenem, and has a broad antibacterial spectrum. This product is expected to be marketed in the near future (EP 0126587; Antimicrobial Agents and Chemotherapy vol. 33, No. 7, 1009(1989). Further Lederle has synthesized biapenem which shows improved antibacterial activities and stability against DHP-I [EP 028801, Antimicrobial agents and chemotherapy vol 35, No. 1, 203 (1991)].

0-Lactam antibiotics are specific for bacteria and show little or no toxic effects against animal cells. Therefore, they are widely used in treatment of infectious disease caused by bacteria. Carbapenem compounds have a broad antibacterial spectrum against gram positive and gram negative, and lower toxicity than other antibacterial agents. However, imipenem resistant bacteria have been clinically isolated and they pose serious problem for the treatment of the disease.

Carbapenem derivatives having an aminocarbonylvinyl group or subsituted or nonsubstituted amino loweralkyl group at the 2 '-position of pyrrolidine whose 4' position is linked as S( sulfur) to the carbapenem skeleton are known in the art. Representative compounds of these classes are (1R,5S,6S)-2-[(2S,4S)-2-[(E)-2-aminocarbonyl)vinyl]pyrrolidin-4-ylthio]-6-[(R)-1-hydroxyethyl]-1-methyl-1-carbapen-2-em-3-carboxylic accd [Compound of Example 3 in EP 0411664; hereinafter referred to simply as BO-2171] and (1R, 5S, 6S)-2- [(2S,4S)-2-[(Z)-3-amino-1-propenyl]-pyrrolidin-4-ylthio]-6-[(R)-1-hydroxyethyl]-1-methyl-1-carbapen-2-em-3-carboxylic acid [Compound of Example 2 in WO 93/01192; hereinafter referred to simply as BO-A]

According to the above references, BO-A is reported to have strong antibacterial activity and to be more stable than BO-2171. Therfore we have selected the BO-A as a reference compound, in addition to imipenem and meropenem, and compared our newly synthesized compounds with these comparative references. The present invention describes structures which contain halo-loweralkenyl, loweralkylsulfonylamino, loweralkysulfamoylamino, side chains attached to 2 position of the pyrrolidine ring of the lactam nucleus, and processes for preparing these compounds. All compounds of these categories have not been introduced or reported in any periodicals or patents.

Extensive research has been directed at identifying compounds with strong antibacterial actvities against various gram positive and gram negative bacteria, including imipenem-resistant strains and clinical isolates of Pseudomonas aeruginosa. which exhibit stability against DHP-1. The present invention describes derivatives with loweralkanesulfonylamino, loweralkylsulfamoylamino side chain attached to the 2 position of pyrrolidine ring of the lactam nucleus exhibit the following properties:

1) strong antibacterial activity against various gram positive and negative strains including beta-lactamase producing strains ; 2) 2-4 times the active against imipenem resistant strains as well as clinical isolates of Pseudomonas aeruginosa than meropenem and BO-A ;

3) increased stable against DHP-1 than meropenem ;

4) more favorable pharmacokinetic properties such as longer half-life, higher under the curve (AUC), higher urinary excretion than imipenem, meropenem, and BO-A ;

5) much lower mortality and toxic effects on the central nervous system than imipenem.

Summary of the Invention

The present invention provides a compound of formula (I)

wherein R₁ is a hydrogen atom or a methyl group, R₂ is a hydrogen atom, or a metal or a nonmetal salt group, or a carboxy protecting group or a negative charge.

The metal or nonmetal salt group of the general formula (I) represents an alkali metal salt such as sodium salt or potassium salt; an alkaline earth metal salt such as magnesium salt or calcium salt; an ammonium salt; an salt composed of an organic base such as triethylamine salt, pyridine salt, ethanolamine salt; or intermolecular quaternary salt. The carboxy protecting group may, for example, be a lower alkyl ester group or an esterified carboxyl group which is mentioned below. The above-mentioned ester group includes at least one appropriate substituent, for example, a lower alkanoyloxy(lower)alkyl ester group such as an acetoxymethyl ester group, a butyryloxymethyl ester group, a pivaloyioxymethyl ester group; a lower alkanesulfonyl(lower)alkyl ester group such as a 2-methylethyl ester group; a lower alkoxycarbonyloxy (lower)alkyl group such as a methoxycarbonyloxymethyl ester group, an ethoxycarbonyloxymethyl ester group, a propoxycarbonyloxymethyl group, a t-butoxycarbonyloxy methyl group; a lower alkenyl group such as a vinyl ester group, an allyl ester group, an isopropenyl ester group; a lower alkynyl group such as an ethynyl ester group, propynyl ester group; an ar( lower) alkyl group capable of having at least more than one substituent such as a benzyl ester group, a 4-methoxybenzyl ester group, a 4-nitrobenzyl ester group, a 2-nitrobenzyl ester group, a trityl ester group, a benzhydryl ester group; an aryl ester group capable of having at least more than one substituent such as a phenyl ester group, a 4-chlorophenyl ester group, a tolyl ester group, and a t-butylphenyl ester group.

"Lower" means that the number of carbon is 1 to 6. The "Lower alkyl" includes a linear or branched alkyl such as methyl, ethyl, propyl, isopropyl, butyl, isobutyl, t-butyl, pentyl, and hexyl. "Lower alkoxy" also includes a linear alkoxy or branched alkoxy such as methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, sec-butoxy, t-butoxy.

R₃ represents a hydrogen atom, an imino protecting group or a pharmaceutically acceptable salt. An appropriate "imino protecting group" may be a carbamoyl, an aliphaticacyl, an aromaticacyl, heterocyclicacyl, an aliphaticacyl substituted with an aromatic group, an aliphaticacyl substituted with a heterocyclic group, all of which are derived from a carboxylic acid, a carbonic acid, and a carbamic acid.

And "a pharmaceutically acceptable salt" may be a salt which has attached hydrobromic acid, hydrochloric acid and sulfuric acid etc.

An appropriate "imino protecting group" may be, for example, (C₂-C₄) alkenyloxycarbonyl, phenyl (C₁-C₄)alkoxy carbonyl, o-niro (or m-nitro or p-nitro) benzyl oxycarbonyl and o-methoxy (or m-methoy or p-methoxy) benzyloxycarbonyl group.

R₄ and R₅ independently represent a hydrogen atom, hydroxy group, cyano group, halogen atom, such as chlorine, bromine, iodine, and fluorine; methanesulfonyl group, N-methylsulfamoyl group, N,N-dimethylsulfamoyl group, lower alkyl group which has a substituent, or no substituent and carbamoyl group which has a substituent or no substituent.

A represents a halogen atom such as chlorine, bromine, iodine and fluorine, or one of the following general formula (1)-(2):

wherein R₆ and R₇ respectively are selected from the group consisting of a hydrogen atom or a lower alkanesulfonyl group, a N-(lower)alkylsulfamoyl group, or a N,N-(lower)dialkyl sulfamoyl group, wherein either of R₆ or R₇ is hydrogen, the other group is a nonhydrogen group.

wherein R₆ is the same as defined above and R₈ is selected from the group consisting of a carbamoyl group which may be optionally substituted with appropriate substituent, or heterocyclic group of 5 or 6 membered ring consisting of 1 to 4 heteroatoms such as a nitrogen atom or an oxygen atom or a sulfur atom which may be optionally substituted with an appropriate substituent;

5- to 6- membered heterocyclic group containing 1 to 4 hetero atoms such as a nitrogen atom, an oxygen atom or a sulfur atom which may be optionally substituted by appropriate substituent, or may be an unsaturated 5- to

6- membered heteromonocyclic group containing 1 to 4 nitrogen atoms, for example, pyrrolyl, pyrrolidinyl, imidazolyl, imidazolidinyl, pyrazolyl, pyrazolidinyl, pyridyl, pyridinio, dihydropyridyl, tetrahydropyridyl ( such as 1,2,3,6-tetrahydropyridyl), pyrimidinyl, pyrimidinio, pyrazinyl, pyrazinio, pyridazinyl(such as 1,3,5-triazinyl, 1,2,4-triazinyl and 1,2,3-triazinyl), tetrahydrotriazinyl(such as 1,2,5,6- tetrahydro-1, 2,4-triazinyl, 1,4,5,6-tetrahydro-1,2,4- triazinyl), triazinio, triazoiyl(such as 1H-1,2,4- triazolyl, 1H-1,2,3-triazolyl, and 2H-1,2,3-triazolyl), triazolio, tetrazinyl, tetrazinio, tetrazoiyl(such as 1H-tetrazolyl and 2H-tetrazolyl), tetrazolio; an unsaturated 5- to 6- membered heteromonocyclic group containing 1 to 2 sulfur atoms and 1 to 3 nitrogen atoms, for example, thiazolyl, thiazolio, isothiazoiyl, thiadiazolyl (such as 1,2,3-thiadiazoyl, 1,2,4-thiαdiαzoiyl, 1,3,4-thiadiazolyl, 1,2,5-thiadiazolyi), thiadiazolio, thiazolydinyl, dihydrothiazynyl; an unsaturated 5- to 6- membered heteromonocyclic group containing 1 to 2 oxygen atoms and 1 to 3 nitrogen atoms, for example, oxadiazolyl (such as 1,2,4-oxadiazoiyl, 1,3,4-oxadiazoiyl, and 1,2,5-oxadiazoiyl).

The above-mentioned heterocyclic group can be substituted with 1 to 3 substituents selected from the group consisting of an amino group, an amino protecting group which is the same as the imino protecting group defined above, a lower alkylamino, carbamoyl and a halo lower alkyl, wherein n is 0 or an integer of 1 to 3.

Also the present invention provides a process for preparing the compound of the formula (I), which comprises reacting a compound of the formula (II) ;

wherein R₁ and R₂ are the same as defined above, R₉ is a hydrogen or a hydroxyl protecting group, with a compound of the formula (III),

wherein R₃, R₄, R₅, A and n are the same as defined above, to obtain a compound of the formula (IV)

wherein R₁, R₂, R₃, R₄, R₅, R₉, A and n are the sames as defined above. The compound of the formula (I) also can be produced by removing any protecting group of the compound of formula (IV), if necessary.

Further, the present invention provides antibacterial compositions comprising an antibacterially effective amount of the compound of formula (I) (wherein R₁, R₂, R₃, R₄, R₅, A and n are the same as defined above) or a pharmaceutical acceptable salts, or ester thereof, and a pharmaceutically acceptable carrier or diluent.

The compound of the present invention has the basic structure as follows:

which is systematically referred to as 7-oxo-1-azabιcyclo[3,2,0]hept-2-ene-2-carboxylιc acid or 1-azabιcyclo[3,2,0]hept-2-ene-7-one-2-carboxylιc acid. In this specification, this basic structure will be referred to as a 1-carbapen-2-em-3-carboxylic acid, by putting the numbers based on a commonly used carbapenom of the following formula;

The present invention includes optical isomers based on the asymmetrical carbon atoms at the 1-position, 5-position, 6-position and 8-position of the carbapenem structure. Among these isomers, preferred is a compound of a (5R,6R,8R) configuration, i.e., a compound having the same steric-configuration (5R,6S) (5,6-trans) as thienamycin in which the carbon atom at the 8-position takes an R-configuration, or a compound of a (1R,5S,6S,8R) configuration in a case where a methyl group is present at the 1-position.

Accordingly, among compounds of the formula (I), a group of compounds having preferred steric configurations are represented by the formula (I-a):

wherein R₁, R₂, R₃, R₄, R₅, A and n are as defined above.

The 2'-substituted pyrrolidin-4'-yl-thio group in the side chain at the 2-position also includes all prossible isomers based on the asymmetrical carbon atoms at the 2- and 4-positions of the pyrrolidine structure. Among these isomers, preferred are compounds of a (2'S,4'S) configuration and a (2'R,4'S) configuration. Among the compounds of formula (I-a) are a group of compounds wherein R₁ is a methyl group, R₄ is hydrogen atom, R₅ is hydrogen atom, methyl group or halogen atom and A is loweralkanesulfonylamino group or N-(lower) alkylsulfamoylamino group which have excellent antibacterial activities.

Further, with respect to the double bond of the 2-substituted pyrrolidine group, cis(Z) and trans (E) geometric isomers are preferred. These isomers are also included in the present invention. Of these isomers, the

(E)-isomer has excellent antibacterial activities.

It is desired that the compound of formula (II) is converted to its reactive derivatives before reacting with the compound of formula (III). That is, the compound of formula (II) is added to the inert organic solvent and reacted with activating agents under alkali conditions to obtain the activated derivatives of the formula (II-a),

wherein R₁, R₂, R₉ is the same as defined above and B is a leaving group. The activating reagent to be used for the reaction may, for example, be an acid anhydride such as methanesulfonic anhydride, trifluromethanesulfonic anhydride, p-toluenesulfonic anhydride, and trifluoroacetic anhydride; or an acid chloride such as methanesulfonyl chloride, p-toluenesulfonyl chloride or diphenyl chlorophosphate. Particulary preferred is diphenyl chlorophosphate.

In the formula (II-a), B is a leaving group such as a trifluoroacetoxy group, a methanesulfonyloxy group, a trifluoromethane.sulfonyloxy group, a p-toluenesulfonyloxy group or a diphenoxyphosphoryloxy group. Particularly preferred is a diphenoxyphos phoryloxy group. The inert organic solvent to be used for the reaction may, for example, be methylene chloride, chloroform, carbon tetrachloride, dichloroethane, trichloroethylene, diethyl ether, tetrahydrofuran, dioxane, benzene, toluene, chlorobenzene, acetone, ethyl acetate, acetonitrile, N,N-dimethylformamide, hexamethylphosphoric triamide or a mixture of such solvents. Particulary preferred is acetonitrile, benzene, toluene, a mixture of toluene and benzene, or a mixture of toluene and ethyl acetate.

The base to be used for the reaction may, for example, be trimethylamine, triethylamine, N,N-diisopropylethylamine, N-methylmorpholine, N-methylpyrrolidine, N-methylpiperidine, N,N- dimethylaniline, 1,8-diazabicyclo[5,4,0]endec-7-ene(DBU), or 1,5-diazabicyclo[4,3,0]-non-5-ene(DBN), or pyridine 4-dimethylaminopyridine, picoline, lutidine, quinoline or isoquinoline. Particularly preferred is N,N-diisopropylethylamine and triethylamine.

For the reaction, from 1 to 3 mol, preferably from 1 to 1.5 mol, of the base and from 1 to 1.3 mol of the activating reagent are used per mol of the compound of formula (II). Though the reaction temperature is not critical, the reaction is conducted usually within a temperature range of from -40° to 50°C, preferably -20° to 20°C, and usually completed quantitatively in from 0.5 to 3 hours. After completion of the reaction, the reaction product is treated in accordance with a conventional method to obtain the reactive derivative (II-a) quantitatively. The compound of formula (II-a) may be reacted with the compound of formula (III) with or without isoiation. The reaction is conducted using the above-mentioned inert organic solvent and the base and from 1 to 2 mol, preferably from 1 to 1.5 mol, of the base and from 0.8 to 1.2 mol of the compound of formula (III) are used per mol of the compound of formula (Il-a).

The reaction is conducted usually within a temperature range of -40° to 50°C, preferably from -20° to 20°C, and usually completed quantitatively in from 0.5 to 10 hours.

Further, the compound of the formula (IV) can be prepared in one pot reaction from the compound of formula (II), namely, without isolating the reactive derivative of formula (Il-a). In the same reaction system, the compound of formula (IV) may be produced with high yield by reacting the compound of formula (II- a) with the compound of formula (III). Thus, from 2 to 5 mol, preferably from 2.5 to 3.8 mol of the base is employed per mol of the compound of formula (II). After completion of the reaction, method well known in the art can be used to obtain a crude product of formula (IV), which may be subjected to a reaction for removing a protecting group without purification. However, it is preferred to purify the crude product (IV) by crystallization or by column chromatography on silica gel.

From the compound of formula (IV) thus obtained, a compound of formula (I) can be obtained, if necessary, by removing a protecting group for a hydroxyl group, an amino group or imino group, and a carboxyl group.

For the removal of the protecting groups, the method varies depending upon the type of the protecting groups. However, the removal can be conducted in accordance with methods known in the art, for example, by addition of a solvent for decomposition; by chemical reduction using a salt of an amine, a metal such as zinc amalgam, a chromic compound such as chromium (II) chloride, chromium (II) acetate together with an organic or inorganic acid such as acetic acid, propionic acid, hydrochloric acid, sulfuric acid; or by catalytic hydrogenation using a platinum or palladium compound. For example, in the formula (IV), the protecting group of the hydroxyi group, amino group or the imino group is an aralkyloxycarbonyl group such as a benzyloxycarbonyl group or a p-nitrobenzyloxycarbonyl group, and the protecting group for the carboxyl group is an aralkyl group such as a benzyl group, a p-nitrobenzyl group or a benzhydryl group. Such protecting groups can be removed by catalytic hydrogenation by means of a platinum catalyst such as platinum oxide, platinum wire or platinum black, or a palladium catalyst such as palladium black, palladium oxide, palladium carbon or palladium hydroxide-carbon (Pearlman's catalyst). When the protecting group of the carboxyl group is an allyl group, isopropenyl, such a protecting group can be removed by catalytic hydrogenation using a palladium ligand complex catalyst such as palladium-carbon, palladium black, palladium hydroxide-carbon, palladium

(II) chloride, tetrakis(triphenylphosphine)palladium

(0), bis(dibenzylidenylacetone)-palladium (0), di(1,2-bis(diphenylphospino)ethane)palladium (0), tetrakis

(triphenylphophine)palladium (II) acetate, di (triphenyl phophine) platinum (II) chloride and the like.

The solvent useful for the catalytic hydrogenation includes, for example, water, methyl alcohol, ethyl alcohol, propyl alcohol, tetrahydrofuran, dioxane, acetonitrile, acetic acid and the above described organic solvent with water, a phosphate buffer(pH=6.5 to 7.0) or 3-(N-morpholino)propanesulfonic acid(MOPS) solution (pH=6.5 to 7.2). The reaction can be completed in from 0.5 to 8 hours at a temperature within a range of from 0° to 40 °C under a hydrogen gas stream of from 1 to 3 atm.

In formula (IV), when the hydroxy group, the amino group, the imino protecting group is an allyloxycarbonyl group, and the protecting group for the carboxyl group is an allyl group, such protecting group can be removed by reacting an organo-soluble palladium ligand complex catalyst in an inert organic solvent containing an allyl group capturing agent (J. Org. Chem., vol 47, 587, 1982).

A solvent useful for the reaction includes, for example, acetone, diethyl ether, tetrahydrofuran, dioxane, ethyl acetate, acetonitrile, methylene chloride, chloroform and a solvent mixture thereof. The allyl group-capturing agent may be, for example, sodium 2-ethylhexanoate, potassium 2-ethylhexanoate, pyridine and piperidine.

The reaction is conducted usually within a temperature range of from -10° to 50°C, preferably from 0° to 30 C° using from 0.01 to 0.5 mol of the palladium ligand complex catalyst and from 0.5 to 5 mol of the nucleophilic agent relative to 1 mol of the compound of the formula (IV), and the reaction is completed usually in from 0.5 to 5 hours.

After completion of the reactions for removing the protecting group, the compound of formula (I) can be isolated by column chromatography on silica gel or adsorptive resin, such as Diaion HP-20, freeze drying or crystallization.

The compound of formula (II) as the starting material, can be obtained by the Salzmann method (J. Am. Chem. Soc. Vol 102, 6161-6163, 1980) in the case where R, is a hydrogen atom, and by the Shih method (Heterocycles, Vol. 21, 29-40, 1984 or EP 0272,455) in the case where R₁ is a methyl group. The compound of formula (III) as the starting material, can be obtained by the following scheme I or Scheme II. In the case of using scheme II, the starting material can be obtained by methods well known to these skilled in the art.

Compound (8) obtained by activation of the hydroxy group of the compound (3) in accordance with methods well known in the art is reacted with potassium thioacetate to convert to an acetylthio derivative (9), followed by alkali or acid hydrolysis to obtain a thiol derivative of formula (III) by Scheme I.

wherein R₃, R₄, R₅, A and n are the same as defined abovo and R₉ in a hydrogen atom or a hydroxy protect ing group and R₁₀ is lower alkyl group and L is a methanesulfonyloxy group, trifluoro acetoxy group, trifluoro methanesulfonyloxy group or p-toluenesulfonyloxy group, and Ac is an acetyl group.

Compound (3) can be prepared in accordance with methods known in the art. Compound (6) can be prepared in accordance with the methods described in the reference examples described below.

The alkene derivative (6) can be obtained by reacting the compound (3) or scheme II with the compound (10), ylide, and the resulting compound (6) may be subjected to the same method as Scheme I to obtain the thiol derivatives of formula (III).

The compound (10) can be prepared by the following method.

wherein R₃, R₄, R₅, R₉, A, L and n are the same as defined above. Compound (11) is a commercially available or prepared by methods known in the art. Abbreviation used are as follows:

AC : acetyl

All : allyl

Alloc : allyloxycarbonyl

Et . ethyl

Me : methyl

Ms : methanesulfonyl

TBS : t-butyldimethyisilyl

Ph : phenyl

E : trans

Z : cis

I. Antibacterial Tests

The antibacterial activities of the compounds of present invention were tested by an agar dilution method in accordance with the standard method of the Japan Chemotherapy Society [Chemotherapy, Vol. 29,76-79 (1981)] which is incorporated herein by reference. After 10 ml of Mueller Hinton Broth was poured into sterilized test tubes, one platinum loopful of each test micro organism was inoculated and incubated overnight at 37°C. Staphylococcus aureuε was cultured in Trypticase Soy Broth instead of Mueller Hinton Broth. The antibacterial agent solutions were prepared by dissolving 5 to 10 mg of each compound in sterilized distilled water to a final the concentration of 1 mg/ml, and by preparing a two-fold serial dilution to concentration of 0.25 μg/ml. After one ml of each of the antibacterial solutions was poured into separate petri dish and sterilized, 9 ml of Mueller Hinton agar media which had been sterilized and cooled was added, mixed well, and solidified to prepare the agar plate media the for Minimal Inhibitory Concentration (MIC; μg/ml) test. 0.11 ml of bacterial culture was poured into a sterilized test tube containing 10 ml of buffered saline gelatin (BSG) soiution and thoroughly mixed. The agar plate containing the antibacterial agent was then inoculated with a bacterial suspension using a stamp, and cultured at 37°C for 18 hours. After observing the growth of bacteria, MIC was determined to be the lowest drug concentration at which there is no growth. The results are shown in Table 1. Table 1. Minimum Inhibitory Concentration (MIC, μg/ml)

Test

Microorgani sm Ceftazidime Imipenem Meropenem BO-A <1-2> <4-2> <5 2> <62>

S.aureus Smith 12.5 ≤0.025 0.1 ≤0.025 ≤.0.025 ≤0.025 ≤0.025 ≤0.025

S.aureus 503 25 ≤0.025 0.1 ≤0.025 ≤0.025 ≤0.025 ≤0.025 ≤0.025

S.pyogenes 308 A 0.39 ≤0.025 ≤0.025 ≤0.025 ≤0.025 ≤0.025 ≤0.025 ≤0.025

E. coli ATCC 9637 0.39 0.2 ≤0.025 0.1 ≤0.025 ≤0.025 0.05 ≤0.025

K.pneumoniae ATCC10031 0.1 0.1 ≤0.025 0.1 ≤0.025 ≤0.025 0.05 0.05

E. cloacae P99 >100 0.78 0.1 0.78 0.2 0.2 0.2 0.39

E.aerogenes B1 >100 100 3.13 12.5 3.13 3.13 6.25 6.25

S.marcescens US12 >100 50 6.25 25 3.13 3.13 6.25 12.5

P.mirabilis ATCC 14273 0.1 3.13 0.1 1.56 0.1 0.1 0.1 0.1

P.aeruginosa DR 1252 1.56 0.39 0.39 0.39 0.05 0.39 0.1 0.39

P.aeruginosa DR 1253 50 6.25 6.25 6.25 1.56 6.25 3.13 6.25

P.aeruginosa DR 1255 25 3.13 1.56 3.13 0.78 3.13 1.56 6.25

P.aeruginosa DR 1260 50 1.56 0.78 1.56 0.2 3.13 0.39 3.13 The compounds of the present invention have exelient antibacterial activity against various gram positive and gram negative bacteria, including beta-lactamase producing strains and imipenem resistant strains. Example <1-2> was the most active of the compounds tested against Psudomonas aeruginosa strains.

II. Porcine Renal dehydropeptidase-I suceptiblity 300g of the swine kidney was homogenized and suspended in a Tris buffer (10mM Tris, 50mM NaCl, 0.01mM ZnCl₂, pH 8.0), and 10% Triton X-100 was added and stirred at 4°C for 24 hours to extract DHP-I. A supernatant, prepared by ultracentrifugation of the above extract at 186,000 × g for 1 hour, was diafiltered to remove Triton X-100. The diafiltered solution was brought to 75% saturation of ammonium sulfate, the resultant precipitate was collected by centrifugation. The above precipitate was dissolved in Tris buffer, and loaded on DEAE-Sepharose fast flow, and anion exchange chromatography was carried out to give the swine DHP-I. The above DHP-I was divided into lml portion at a concentration of 1 unit/ml and stored at -70°C.

Glycyldehydrophenylalanine (GDP), imipenem, meropenem and BO-A were employed as control compounds. As shown in Table 2, the compounds of the present invention have higher stability to DHP-1 than imipenem and meropenem.

III. Pharmacokinetics in mice

A test compound was dissolved in sterillized distilled water, and a dose of 20mg/kg of body weight was inoculated under the skin of ICR mice weighing 20 ± 2g.

10 μl of blood samples was collected from the tail vein at preset sampling interval. Urine was collected for 18 hours from another group of mice (n=10) which were housed in metbolism cages. Bioassays were carried out to measure the concentration of a blood and urine by using Bacillus subtilis ATCC 6633 as a test bacterial strain.

As shown in Table 3, compound of the present invention, example 1-2, has longer half-life and higher area under the curve (AUC) than imipenem, meropenem and BO-A and has 1.5 to 3 times higher uninary recovery than the control compounds.

IV. Toxicity in mice 2000, 1500, and 1000 mg/kg of the compounds of present invention were injected into 20±2 g weight ICR mice via the tail vein. After 2 weeks, the mice were observed (n=3). In the case of compounds 1-2 and 5-2, central nervous system (CNS) side effects such as death and jerks were not observed. But in case of imipenem injection at a dosage of 1,500 mg/Kg, one of three of the mice died and all three of the mice showed jerks as soon as they were injected. The compounds of the present invention have excellent antibacterial activities against various gram positive bacteria and gram negative bacteria including beta lactamase producing strains and imipenem resistant strains. Also, the compounds of the present invention showed favorable pharmacokinetic properties, good physicochemical stability and high water slubility. Therfore, the compounds of the present invention are useful as antibacterial agents for treatment and prevention of human infectious diseases caused such bacteria. Because of their broad antibacterial spectrum, the compounds of the present invention may be used in the form of additives for animal food, preserving agents, and other sterilization and disinfection agents for industrial use as well as medical use.

The compound of the present invention may be used in the form of a drug formulation suitable for non-oral administration, oral administration, external administration; a liquid formulation such as an injection solutions, syrups or emulsions; solid formulation such as tablets, capsules or granules; and external application formulations such as ointments or suppositories.

Dosages vary depending upon the condition of the patient, the weight, age, sex, type of formulation, and how the dose is to be administered. Usually, however, a preferred daily dose of the active ingredient to an adult is from about 5 to 50 mg/kg, and a preferred daily dose to a child is within a range of from about 5 to 25 mg/kg, which is preferably administered once a day or several times a day.

The compound of present invention has high stability to renal DHP-I, therefore, they may be administered by themselves, and if necessary, may be administered in combination with a DHP-I inhibitory agent such as cilastatin. The present invention will now be described in further detail with reference to Examples and Reference

Examples. However, it should be understood that the present invention is by no means restricted by such specific examples.

In the Examples and Reference Examples, for the thin layer chromatography, silica gel 60 F₂₅₄ (Merck) was used as the thinlayer plate material, and an ultraviolet detector or ninhydrin or KMnO₄ color development method was used as a detection means. For columns, silicagel 60 (Merck) was used, and UV spectrophotometer DMS 100S (Varian) was used for detecting the UV absorbency. For high speed liquid chromatography measurement a M-352 (ACS) model was used. When the NMR spectrum was measured using dimethyl sulfoxide-d₆ or chloroform-d, solution, tetramethylsilane (TMS) was used as the internal standard material, and when measured using a deuterium oxide solution, 2,2-dimethyl-2-silapentane-5-sulfonate (DSS) was used as the internal standad. The measurement was conducted by means of AC 200P (200 MHzrBruker) model spectrometor. All δ values are represented by ppm.

The meaning of the abbreviation used in NMR measurement are as follows : s : singlet bbdd :: broad doublet d : doublet

CDCl₃ : chlorform-d.

DMSO-d₆ : dimethylsulfoxide-d. 6

t : triplet D₂O : deuterium oxide q : quartet J : coupling constant br : broad Hz : hertz

dd : double doublet

m : multiplet

bs : broad singlet REFERENCE EXAMPLE 1

(2S,4S)-1-allyloxycarbonyl-2[(E)-3-methanesulfonyl amino-1-propenyl]-4-mercaptopyrroiidine

REFERENCE EXAMPLE 1-1

To a solution of 33 g (94.7 mmol) of ethyl tri(phenyiphosphoranylidene) acetate in 100 ml of methylene chloride was added dropwise a solution 15 g (47.7 mmol) of (2S,4R)-1-allyloxycarbonyl-2-formyl-4-t-butyldimethylsilyloxypyrrolidine obtained by known processes in 100 ml of methylene chloride at room temperature. The mixture was stirred for 3 hours, and concentrated under reduced pressure. 500 ml of a mixture of ethyl acetate and hexane (1:4 v/v) was added, stirred for 10 minutes, filtered and concentrated under reduced pressure. The residue was purified by column chromato graphy on silica gel (eluted with ethyl acetate: hexane = 1:8) to give 17.15 g (95% yield) of ethyl 3-[ (2S,4R)-1-allyloxycarbonyl-4-t-butyldimethylsilyloxypyrrolidin-2-yl]acrylate.

NMR (CDCl₃) δ:

0.01(s,6H), 0.82(S,9H), 1.20-1.27 (t,3H), 1.80- 1.86(m,1H), 2.03-2.07(m,1H), 3.32-3.48 (m,2H), 4.08- 4.19(q,2H), 4.29-4.36 (m,1H), 4.51-4.54 (m,3H), 5.14- 5.29(m,2H), 5.82-5.89(m,2H), 6.73-6.84 (dd, 1H)

REFERENCE EXAMPLE 1-2

To a solution of 11 g (29.06 mmol) of ethyl 3- [(2S,4R)-1-allyloxycarbonyl-4-t-butyldimethylsilyloxy pyrrolidin-2-yl] acrylate obtained in REFERENCE EXAMPLE

1-1 in 100 ml of tetrahydrofuran, 84.28 ml (84.27 mmol) of 1.0 M diisobutylaluminum hydride solution in hexane was added dropwise at -70°C. The mixture was stirred for 30 minuites at -70°C. Then, the temperature was raised to room temperature. To the reaction solution, 200 ml of ethyl acetate and 20 ml of methanol was added, and the mixture was stirred for another 1 hour. The precipitate which formed was collected by filtration and the filtrate was concentrated under reduced pressure. The residue was purified by column chromatography on silica gel (eluted with ethyl acetate: hexane = 4:1) to give 6.64 g (69 % yield) of (2S,4R)-1-allyloxycarbonyl-2-[(E)-3-hydroxy-1-propenyl]-4-t-butyldimethylsilyloxy pyrrolidine. NMR (CDCl₃) δ:

0.03(s,6H), 0.84(s,9H), 1.67-1.71 (m, 2H), 3.41- 3.71(m,3H), 4.10-4.12(d,1H), 4.32-4.36 (m, 2H), 4.52- 4.58(m,3H), 5.21-5.30(m,2H), 5.62-6.61 (m, 2H)

REFERENCE EXAMPLE 1-3

After 6.64 g (19.74 mmol) of (2S,4R)-1-allyl oxycarbonyl-2-[(E)-hydroxyl-1-propenyl]-4-t-butyl dimethylsilyloxypyrrolidine obtained in REFERENCE EXAMPLE 1-2 was dissolved in 65 ml of tetrahydrofuran, 6.21 g (23.69 mmol) of triphenylphosphine and 3.48 g (23.69 mmol) of phthalimide were added to the reaction mixture and cooled in an ice bath. Then, to the mixture, 3.73 ml (23.69 mmol) of diethyi azodicarboxy late was added dropwise. The mixture was stirred for 30 minutes, and then concentrated under reduced pressure. The resulting residue was dissolved in ethyl acetate. The mixture was washed with distilled water and then a saturated sodium chloride agueous solution. The organic layer was dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified by column chromatography on silica gel (eluted with ethyl acetate:hexane = 1:8) to give 8.8 g (95 % yield) of (2S,4R)-1-allyloxycarbonyl-2-[(E)-3-phthalimido-1-propenyl]-4-t-butyldimethylsilyloxy pyrrolidine.

NMR (CDCl₃) δ: 0.07(s,6H), 0.81(s,9H), 1.70-1.82 (m, 1H), 1.96- 2.01(m,1H), 3.41-3.47(m,2H), 4.22-4.33 (m, 3H), 4.46- 4.53(m,4H), 4.81-5.28(m, 2H), 5.62-5.80 (m, 2H), 7.68-7.73(m,2H), 7.77-7.84(m,2H) REFERENCE EXAMPLE 1-4

To a solution of 8.8 g (18.70 mmol) of (2S,4R)-1-allyloxycarbonyl-2-[(E)-3-phthalimido-1-propenyl]-4-t-butyldimethylsilyloxypyrrolidine prepared in REFERENCE EXAMPLE 1-3 in 90 ml of ethanol, 2.09 ml (43.08 mmol) of hydrazine hydrate was added. The mixture was stirred for 12 hours, filtered and the filtrate was concentrated under reduced pressure. The residue was diluted with ethyl acetate. The mixture was washed in sequence with a 3N-ammonia solution, distilled water and saturated sodium chloride aqueous solution, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was diluted with 100ml of dichloromethane. To the mixture, 3.13 ml (22.44 mmol) of triethylamine and 1.8 ml (22.31 mmol) of methanesulfonyl chloride were added and cooled on an ice. The reaction mixture was stirred for 30 minutes, washed in sequence with 1N-HCl, saturated sodium carbonate solution, distilled water, and saturated sodium chloride aqueous solution, dried over anhydrous sodium sulfate, and then concentrated under reduced pressure. The residue was purified by column chromatography on silica gel (eluted with ethyl acetate: hexane = 1:2) to give 6.26g (80 % yield) of (2S,4R)-1-allyloxycarbonyl-2-[(E)-3-methanesulfonyl-1-propenyl]-4-t-butyldimethylsilyloxypyrrolidine.

NMR (CDCl₃) δ:

0.03(s,6H), 0.84(S,9H), 1.66-1.78 (m, 1H), 1.98-2.07(m,1H), 2.93(S,3H), 3.44-3.46 (d, 2H), 3.70-3.74 (m,2H), 4.30-4.44(m,2H), 4.52-4.56 (m,3H), 5.14-5.30 (m,2H), 5.62(bs,2H), 5.81-5.98 (m, 1H)

REFERENCE EXAMPLE 1-5 To a solution of 6.26 g (14.95 mmol) of (2S,4R)-1-allyloxycarbonyl-2-[(E)-3-methanesulfonyl-1-propenyl]-4-t-butyldimethylsilyloxypyrrolidine prepared in REFERENCE EXAMPLE 1-4 in 80 ml of tetrahydrofuran, 5.8 g (22.18 mmol) of tetrabutylammonium fluoride hydrate was added. The mixture was stirred for 1.5 hours. After concentration under reduced pressure, the residue was dissolved in chloroform, washed with distilled water and then saturated sodium chloride aqueous solution and dried over anhydrous magnesium sulfate, and concentrated under reduced pressure. The residue was dissolved in 80 ml of dichloromethane. To the mixture, 2.5 ml (17.92 mmol) of triethylamine and 1.45 ml (17.92 mmol) of methanesulfonyl chloride were added and cooled on an ice. The reaction mixture was stirred for 30 minutes, washed with 1N-HCl, distilled water, saturated sodium carbonate solution, and distilled water, dried over anhydrous magnesium sulfate and concentrated under reduced pressure to give 4.3 g (75 % yield) of (2S,4R)- 1-allyloxycarbonyl-2-[(E)-3-methanesulfonylamino-1- propenyl]-4-methanesulfonyloxypyrrolidine. NMR ( CDCl₃) δ :

1.75(m,1H), 2.01(m,1H), 2.93(s,3H), 3.02(s,3H), 3.62-3.74(m, 3H) , 3.82-3.94 (m, 1H) , 4.48-4.56 (m, 3H), 5.16-5.31(m,3H), 5.62-5.64 (m, 2H), 5.81-5.97 (m, 1H)

REFERENCE EXAMPLE 1-6

To a solution of 4.3 g (11.24 mmol) of (2S,4R)-1-allyloxycarbonyl-2-[(E)-3-methanesulfonylamino-1-propenyl]-4-methanesulfonyloxypyrrolidine prepared in REFERENCE EXAMPLE 1-5 in 150 ml of anhydrous acetonitrile, 1.9 g (16.6 mmol) of potassium thioacetate was added. The mixture was heated to reflux for 4 hours. After cooling in an ice bath, insoluble material was removed by filtration. The filtrate was concentrated under reduced pressure. The residue was dissolved in ethyl acetate. The residue was washed with distilled water, dried over anhydrous sodium sulfate and concentrated under reduced pressure. The residue was purified by column chromatography on silica gel (eluted with ethyl acetate: hexane = 1:1) to give 3.26 g (80 % yield) of (2S,4S)-1-allyloxycarbonyl-2-[(E)-3-methanesulfonyl amino-1-propenyl ]-4-thioacetylpyrrolidine.

NMR ( CDCl₃) δ:

2

0462 -1.82(m,1H), 2.31(m,1H), 2.52-2.61 (m, 1H ) , 2.94(s,3H), 3.23-3.31(m, 1H), 3.71-3.74 (m, 2H), 3.88-

4.04(m,2H), 4.36-4.40(m, 1H), 4.51-4.55 (m, 2H), 5.16-

5.31(m,2H), 5.64-5.67(bd,2H), 5.78-5.98 (m, 1H)

REFERENCE EXAMPLE 1-7

To a solution of 3.26 g (8.99 mmol) of (2S,4S)-1- allyloxycarbonyl-2-[(E)-3-methanesulfonylamino-1-propenyl]-4-thioacetylpyrrolidine prepared in REFERENCE EXAMPLE 1-6 in 50 ml of methanoi, 14.25 ml (28.5 mmol) of 2 N-NaOH was added under ice cooling. The mixture was stirred for 3 minutes. It was neutralized with aqueous citric acid and concentrated under reduced pressure. The residue was purified by column chromatography on silica gel (eluted with ethyl acetate: hexane = 1:1) to give 2.45 g (85 % yield) of (2S,4S)-1-allyloxycarbonyl-2-[(E)-3-methanesulfonylamino-1-propenyl]-4-mercaptopyrrolidine.

NMR ( CDCl₃) δ: 1.60-1.78 (m, 1H), 1.69-1.73 (d, 1H), 2.54-2.61 (m, 1H),

2.94(S,3H), 3.13-3.31 (m, 1H), 3.70-3.76 (m, 2H), 4.01-4.09 (m,1H), 4.28-4.36(m,1H), 4.50-4.55 (m, 2H), 5.16-5.22 (m,2H), 5.61-5.71(m,2H), 5.81-6.00 (m, 1H) REFERENCE EXAMPLE 2

(2S,4S)-1-allyloxycarbonyl-2-[(E)-3-methaneεulfonylamino-1-methylpropenyl]-4-mercaptopyrrolidine REFERENCE EXAMPLE 2-1

To a solution of 1.0 g (25.7 mmol) of sodium hydride (60 %) in 106 ml of tetrahydrofuran, 1.13 g (25.7 mmol) of triethyl phosphonoacetate was added dropwise at 0 °C. 7.0 g (21.4 mmol) of (2S,4R)-1- allyloxycarbonyl-2-methylcarbonyl-4-t-butyldimethylsilyl oxypyrrolidine obtained by known processes, was added dropwise at the same temperature. The mixture was warmed to room temperature and stirred for 15 hours at the same temperature. Ethyl acetate was added in an excess amount. The mixture was washed with aqueous sodium chloride, dried over anhydrous magnesium sulfate, and then concentrated under reduced pressure. The residue was purified by column chromatography on silica gel (eluted with ethyl acetate: hexane = 1:10) to obtain 3.3 g (95 % yield) of (2S,4R)-1-allyloxycarbonyl-2-[(E)-3-ethoxycarbonyl-1-methylpropenyl]-4-t-butyl dimethylsilyloxypyrrolidine, together with the recovery of 4.13g of starting material. NMR (CDCl₃) δ:

0.03(s,6H), 0.83(s,9H), 1.24(t,3H), 1.80(m,1H), 2.05(s,3H), 3.40-3.70 (m, 2H), 4.15(q,2H), 4.31(m,1H), 4.58(m,2H), 5.20(m,2H), 5.73(s,1H), 5.85(m,1H)

REFERENCE EXAMPLE 2-2

To a solution of 3.3 g (8.3 mmol) of (2S,4R)-1-allyloxycarbonyl-2-[(E)-3-ethoxycarbonyl-1-methyl propenyl]-4-t-butyldimethylsilyloxypyrrolidine obtained in reference example 2-1 in 42 ml of tetrahydrofuran, 16 ml (24.1 mmol) of diisobutylaluminum hydride (1.5 M hexane solution) was added dropwise at -78 °C. The mixture was stirred for 1.5 hours at the same temperature. The reaction mixture was warmed to 0 °C.

10 ml of methanol was added dropwise to the mixture.

The mixture was stirred for another about 30 minutes.

Ethyl acetate was added and insoluble material was removed by filtration. Then the mixture was concentrated under reduced pressure. The residue was refined by column chromatography on silica gel (eluted with ethyl acetate: hexane = 1:2) to obtain 1.76 g (60 % yield) of (2S,4R)-1-allyloxycarbonyl-2-[(E)-3-hydroxy-1- methyl propenyl]-4-t-butyldimethylsilyloxypyrrolidine.

NMR (CDCI₃) 0.03(s,6H), 0.84(s,9H), 1.60(s,3H), 1.79(m,1H), 2.00(m,1H), 3.50(m,2H), 4.15(d,2H), 4.30(m,2H),

4.55(m,2H), 5.20(m,2H), 5.22(t,1H), 5.90(m,1H) REFERENCE EXAMPLE 2-3

To a solution of 1.76 g (5.0 mmol) of (2S,4R)-1-allyloxycarbonyl-2-[(E)-3-hydroxy-1-methylpropenyl]-4-t-butyldimethylsilyloxypyrrolidine obtained in REFERENCE EXAMPLE 2-2 in 25 ml of tetrahydrofuran, 1.9 g (7.4 mmol) of triphenylphosphine and 1.3 g (8.7 mmol) of phthalimide were added at 0 °C and 1.2 ml (7.4 mmol) diethyi azodicarboxylate was added dropwise. The mixture was stirred at the same temperature and then concentrated under reduced pressure. The residue obtained therein was refined by column chromatography on silica gel (eluted with ethyl acetate : hexane = 1:5) to obtain 2.37 g (98 % yield) of (2S,4R)-1-allyloxycarbonyl-2-[(E)-3-phthalimido-1-methylpropenyl]-4-t-butyldimethylsilyloxy pyrrolidine.

NMR (CDCl₃) δ :

0.01(s,6H), 0.81(s,9H), 1.73(s,3H), 1.97(m, 1H), 3.48(m, 2H), 4.28(m, 4H), 4.50(m,2H), 5.80(m,1H), 7.67(m,2H), 7.80(m,2H)

REFERENCE EXAMPLE 2-4 To a solution of 2.37 g (4.9 mmol) of (2S,4R)-1-allyloxycarbonyl-2-[(E)-3-phthalimido-1-methylpropenyl]-4-t-butyldimethylsilyloxypyrrolidine obtained in REFERENCE EXAMPLE 2-3 in 40 ml of ethanol, 0.06 ml of hydrazine hydrate was added. The mixture was stirred for 13 hours at room temperature. The insoluble material was removed by filtration and the filtrate was concentrated under reduced pressure. The residue was dissolved in 20 ml of dichloromethane. To the mixture were added 0.4 ml (5.2 mmol) of methanesulfonyl chloride and 0.72 ml (5.2 mmol) of triethylamine at 0 °C. The mixture was then stirred for 30 minites at the same temperature, and washed with water and then aqueous sodium chloride, and dried over anhydrous magnesium sulfate. The organic layer was then concentrated under reduced pressure. The residue was refined by column chromatography on silica gel (eluted with ethyl acetate: hexane = 1:1.5) to obtain 1.4 g (66 % yield) of

( 2S, 4R)-1-allyloxycarbonyl-2-[ (E)-3-methanesulfonylamino-1-methylpropenyl]-4-t-butyldimethylsilyloxypyrrolidine.

NMR ( CDCl₃) δ :

0.03(s,6H), 0.83(s,9H), 1.59(s,3H), 1.79(m,1H), 2.00(m,1H), 2.93(m,3H), 3.50(m,2H), 3.78(t,2H), 4.35(m,2H), 4.51(m,2H), 5.10-5.41 (m, 3H), 5.88(m,1H)

REFERENCE EXAMPLE 2-5

To a solution of 1.4 g (3.2 mmol) of (2S,4R)-1- allyloxycarbonyl-2-[(E)-3-methanesulfonylamino-1-methylpropenyl]-4-t-butyldimethylsilyloxypyrrolidine in

15 ml of methanol, 0.9 ml of 6N-HCl was added. The mixture was stirred for 1.5 hours at room temperature.

The mixture was then neutralized with 2N-NaOH and concentrated under reduced pressure. The residue was extracted with ethyl acetate. The organic layer was washed with water and then aqueous sodium chloride and dried over anhydrous magnesium sulfate and concentrated under reduced pressure. The residue was dissolved in 13 ml of dichloromethane and 0.24 ml (3.1 mmol) of methane sulfonyl chloride and 0.44 ml (3.1 mmol) of triethyl amine were added to the mixture and the mixture stirred for 30 minutes at 0 °C. The reaction mixture was washed with water and then aqueous sodium chloride, dried over anhydrous magnesium sulfate, and concentrated under reduced -pressure to obtain 0.96 g (75 % yield) of

( 2 S , 4R ) -1 -a l lyl oxycarbonyl - 2 - [ ( E ) -3-methanesulfonylamino-1-methylpropenyl ]-4-methanesulfonyloxypyrrolidine. NMR (CDCl₃) δ :

1.61(S,3H), 2.00(m,1H), 2.42(m,1H), 2.93(s,3H), 3.03(s,3H), 3.79(t.2H), 4.50(m,3H), 5.40(t,1H),

5.88(m,1H)

REFERENCE EXAMPLE 2-6

To a solution of 0.96 g (2.4 mmol) of (2S,4R)-1-allyloxycarbonyl-2-[(E)-3-methanesulfonylamino-1-methylpropenyl]-4-methanesulfonyloxypyrrolidine obtained in REFERENCE EXAMPLE 2-5 in 10 ml Of DMF and 10 ml of toluene, 0.55 g (4.8 mmol) of potassium thioacetate were added. The mixture was stirred for 1.5 hours at 90 °C. 300 ml of ethyl acetate was added to the mixture. The mixture was washed with water and then an aqueous sodium chloride, dried over anhydrous magnesium sulfate, and concentrated under reduced pressure. The residue was dissoived in 15 ml of methanol, and 2 ml (4 mmol) of 2N-NaOH was added. The mixture was stirred for 10 minutes, and then neutralized with 1N-HCl and concentrated under reduced pressure. The residue was extracted with ethyl acetate. The organic layer was washed with water and an aqueous sodium chloride, dried over anhydrous magnesium sulfate, and concentrated under reduced pressure. The residue was refined by column chromatography on silica gel (eluted with ethyl acetate: hexane = 1:1) to obtain 0.33 g (41 % yield) of (2S,4S)-1-allyloxycarbonyl-2-[ (E)-3-methanesulfonylamino-1-methylpropenyl]-4-mercaptopyrrolidine. NMR (CDC1₃) δ :

1.58(s,3H), 1.69(d,1H), 2.54(m,1H), 2.94(s,3H), 3.20(m,2H), 3.80(t.2H), 4.58(m,2H), 5.40(t,1H), 5.90(m,1H)

REFERENCE EXAMPLE 3

(2S_,4S)-1-allyloxycarbonyl-2-[(E)-3-fluoro-1-propenyl]-4-mercaptopyrrolidine

REFERENCE EXAMPLE 3-1

To a solution of 1.03 g (3.06 mmol) of (2S,4R)-1-allyloxycarbonyl-2-[(E)-3-hydroxy-1-propenyl]-4-t-butyldimethylsilyloxypyrrolidine obtained in REFERENCE

EXAMPLE 1-2 in 30 ml of dichloromethane, 0.5 ml (3.78 mmol) of diethylaminosulfur trifluoride (DAST) was added dropwise at 0 °C. The mixture was stirred for 30 minutes under a nitrogen stream at the same temperature.

5 ml of saturated sodium carbonate aqueous soltuion was added to the mixture. The mixture was stirred for 10 minutes, and then extracted with chloroform. The organic layer was concentrated under reduced pressure, and refined by column chromatography on silica gel (eluted with ethyl acetate: hexane = 1:5) to obtain 0.35 g (34 % yield) of (2S,4R)-1-allyloxycarbonyl-2-[(E)-3-fluoro-1- propenyl]-4-t-butyldimethylsilyloxypyrrolidine. NMR (CDCl₃) δ : 0.04(s,6H), 0.84(s, 9H), 1.85(m,1H), 2.02(m,1H), 3.47(d,m,2H), 4.34(t,1H), 4.53(m,1H), 4.57(m,2H), 4.70(bs, 1H), 4.94(bd,1H), 5.15(d,1H), 5.21(d,1H), 5.74(bs, 2H), 5.85(m,1H)

REFERENCE EXAMPLE 3-2

0.31 g (0.92 mmol) of (2S,4R)-1-allyloxycarbonyl-2-[(E)-3-fluoro-1-propenyl]-4-t-butyldimethylsilyloxy-pyrrolidine obtained in REFERENCE EXAMPLE 3-1 was treated by the same process as that of REFERENCE EXAMPLE 2-5 to obtain a desired material, 0.26 g (94 % yield) of (2S,4R)-1-allyloxycarbonyl-2-[(E)-3-fluoro-1-propenyl]-4-methanesulfonyloxypyrrolidine.

NMR (CDCl₃) δ :

2.05(m,1H), 2.50(bs,1H), 3.03(s,3H), 3.63(dd,1H), 3.95(bd, 1H), 4.57(m,3H), 4.71(d,1H), 4.95(d,1H), 5.22(m,3H), 5.78(bs,2H), 5.82(m,1H)

REFERENCE EXAMPLE 3-3

0.24 g (0.79 mmol) of (2S, 4R)-1-allyloxycarbonyl-2- [(E)-3-fluoro-1-propenyl]-4-methanesulfonyloxy pyrrolidine obtained in REFERENCE EXAMPLE 3-2 was treated by the same process as that of reference example 1-6 to obtain a desired material, 0.32 g (100 % yield) of (2S,4S)-1-allyloxycarbonyl-2-[(E)-3-fluoro-1-propenyl]-4-acetylthiopyrrolidine.

NMR (CDCl₃) δ :

1.76(m-1H), 2.31(S,3H), 2.57(m,1H), 3.96(m,2H), 4.42(m,1H), 4.55(m,3H), 4.71(d,1H), 4.95(d,1H),

5.19(d,1H), 5.23(d,1H), 5.77(bs,2H), 5.82 (m, 1H) REFERENCE EXAMPLE 3-4

0.31 g (1.1 mmol) of (2S, 4S)-1-allyloxycarbonyl-2-[(E)-3-fluoro-1-propenyl]-4-acetylthiopyrrolidine obtained- in REFERENCE EXAMPLE 3-3 was treated by the same process as that of REFERENCE EXAMPLE 1-7, and the crude product obtained as described above was refined by column chromatography on silica gel (eluted with chloroform: acetone = 8:1) to give 0.12 g (63 % yield) of (2S,4S)-1-allyloxycarbonyl-2-[(E)-3-fluoro-1-propenyl]-4-mercaptopyrrolidine.

NMR (CDCl₃) δ : 1.70(m-1H), 2.60(m,1H), 4.04(bS,2H), 4.38(m,1H),

4.56(m,3H), 4.71(d,1H), 4.95(d,1H), 5.17(d,1H), 5.26(d,1H), 5.80(bs,2H), 5.82(m,1H)

REFERENCE EXAMPLE 4

( 2S,4S)-1-allyloxycarbonyl-2-[(Z)-3-methanesulfonylamino-1-propenyl]-4-mercaptopyrroiidine

REFERENCE EXAMPLE 4-1

41 g (130.8 mmol) of (2S,4R)-1-allyloxycarbonyl-2- formyl-4-t-butyldimethylsilyloxypyrroiidine obtained by known process was dissolved in 400 ml of methanoi and cooled to -20 °C. 23.9 g (69.1 mmol) of (carbetoxy methylene)triphenylphosphorane was added dropwise to the mixture. The reaction mixture was stirred overnight at -20°C, and concentrated under reduced pressure. The residue was dissolved in the mixture solution (1:4 v/v) of ethyl acetate and hexane, and stirred for 10 minutes. The insoluble material was removed by filtration and the filtrate was concentrated under reduced pressure. The residue was refined by column chromatography on silica gel (eluted with ethyl acetate: hexane = 1:8) to give 18. lg (72 % yield) of ethyl 3-[(2S,4R)-1-allyloxycarbonyl-4-t-butyldimethylsilyloxypyrrolidine-2-yl] acrylate.

NMR(CDCl₃) δ : 0.05(S,6H), 1.90(S,9H), 1.30(t,3H), 1.80(m,1H),

2.10(m,1H), 3.50(m,2H), 4.15(q,2H), 4.35(m,1H), 4.59(m,2H), 5.20(m,2H), 5.43 (m, 1HX2/3), 5.84 (m, 1HX7/3), 6.15(m,1Hx2/3), 6.80(dd,1Hx1/3) REFERENCE EXAMPLE 4-2

18 g (46.9 mmol) of ethyl 3-[(2S,4R)-1-allyloxycarbonyl-4-t-butyldimethylsilyloxypyrrolidin-2-yl] acrylate obtained in REFERENCE EXAMPLE 4-1 was dissolved in 250 ml of anhydrous tetrahydrofuran, and cooled to -78 °C using a dry ice -acetone bath. To the reaction mixture, 96.5 ml (144.8 mmol) of 1.5 M diisobutylaluminum hydride was added dropwise and the mixture was stirred for 2 hours at the same temperature and warmed to room temperature. 100 ml of methanoi was added and the mixture was stirred for half an hour at the same temperature. The solid material was removed by using celite. The filtrate was concentrated under reduced pressure, and the residue was refined by column chromatography on silica gel (eluted with ethyl acetate: hexane = 4:1) to give 6 g (60 % yield) of (2S,4R)-1- allyloxycarbonyl-2-[(Z)-3-hydroxy-1-propenyl]-4-t- butyldimethyl silyloxypyrrolidine and 5 g (30 % yield) of (2S,4R)-1-allyloxycarbonyl-2-[(E)-3-hydroxy-1- propenyl]-4-t-butyldimethylsilyloxypyrrolidine, respectively. (Z) form

NMR(CDCl₃) δ : 0.05(S,6H), 0.90(s,9H), 1.70(m,1H), 2.05(m,1H),

3.50(m,2H), 3.90(m,1H), 4.30-4.60 (m, 5H), 5.90(m,1H), 5.30(m,3H), 5.90(m,1H)

(E) form NMR(CDCl₃) δ :

0.05(s,6H), 0.95(S,9H), 1.80(m,1H), 2.01(m,1H), 3.50(m,2H), 4.10(d,2H), 4.30-4.60 (m, 4H), 5.20(m,2H), 5.50-5.95(m,3H)

REFERENCE EXAMPLE 4-3

5.03 g (14.73 mmol) of (2S,4R)-1-allyloxycarbonyl -2-[(Z)-3-hydroxy-1-propenyl]-4-t-butyldimethylsilyloxy pyrrolidine obtained in REFERENCE EXAMPLE 4-2 was dissolved in 150 ml of tetrahydrofuran, and cooled to 0 °C. 5.8 g (22.1 mmol) of triphenylphosphine and 3.8 g (25.8 mmol) of phthalimide were added to the mixture. The reaction mixture was stirred for 10 minutes. Then, to the mixture was added 3.49 g (22.2 mmol) of diethyi azodicarboxylate. The mixture was stirred for another 2.5 hours at 0 °C and concentrated under reduced pressure. To the residue, ethyl acetate and water were added and the mixture was extracted with ethyl acetate. The organic layer was washed with water and then saturated sodium chloride aqueous solution, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was refined by column chromatography on silica gel (eluted with ethyl acetate:hexane = 1:7) to give 4.71 g (70 % yield) of (2S,4R)-1-allyloxycarbonyl-2-[(Z)-3-phthalimido-1-propenyl]-4-t-butyldimethylsilyloxy pyrrolidine.

NMR(CDCl₃) δ :

0.05(s,6H), 0.90(s,9H), 1.80(m,1H), 2.10(m,1H), 3.50(m,2H), 4.30-4.63(m, 5H), 4.90-5.50 ( m, 5H), 5.90(m,1H), 7.60-7.85(m,4H) REFERENCE EXAMPLE 4-4

To a solution of 4.7 g (10.3 mmol) of (2S,4R)-1-allyloxycarbonyl-2-[(Z)-3-phthalimido-1-propenyl]-4-t-butyldimethylsilyloxypyrrolidine obtained in REFERENCE EXAMPLE 4-3 in 250 ml of anhydrσus ethanol, 1.6 ml (47.4 mmol) of hydrazine hydrate was added. The mixture was stirred overnight at room temperature. The solid material was removed by filtration and the filtrate was concentrated under reduced pressure. To the residue, ethyl acetate and water were added, and the organic layer was washed in sequence with 3 N-ammonia solution, saturated sodium chloride aqueous solution, dried over the anhydrous sodium sulfate and concentrated under reduced pressure. The residue was dissolved in 100 ml of methylene chloride and cooled to 0 °C. To the mixture were added 7.2 ml (51.5 mmol) of triethylamine and 2.8 ml (36.1 mmol) of methanesulfonyl chloride, and the mixture was stirred at 0 °C for half an hour. 100 ml of methylene chloride and water were added to the reaction mixture, and the organic layer was washed with a saturated citric acid solution and, then saturated sodium chloride aqueous solution, dried over the anhydrous sodium sulfate and concentrated under reduced pressure. The residue was refined by column chromatography on silica gel (eluted with ethyl acetate: hexane=1:1.5) to give 2.21 g (51.3 % yield) of (2S,4R)- 1-allyloxycarbonyl-2-[(Z)-3-methanesulfonylamino-1-propenyl]-4-t-buryldιmethylsilyloxypyrrolidine.

NMR(CDCl₃) δ :

0.05(S,6H), 0.90(s,9H), 1.70(m,1H), 2.05(m,1H), 2.93(s,3H), 3.40(m,2H), 3.60(m,1H), 4.01(m,1H), 4.30-4.75(m,3H), 5.10-5.40(m,3H), 5.61-5.98(m,3H) REFERENCE EXAMPLE 4-5

To a solution of 2.2 g (5.26 mmol) of (2S,4R)-1-allyloxycarbonyl-2-[(Z)-3-methanesulfonylamino-1-propenyl]-4-t-butyldimethylsilyloxypyrrolidine obtained in REFERENCE EXAMPLE 4-4 in 150 ml of tetrahydrofuran, 6.6 ml (6.6 mmol) of lM-tetrabutylammonium fluoride was added at room temperature. The mixture was stirred for half an hour and concentrated under reduced pressure. To the mixture was added ethyl acetate and water and then organic layer was washed with water and then saturated sodium chloride aqueous solution, dried over the anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was dissolved in 100 ml of methylene chloride and cooled to 0 °C. To the reaction mixture were added 2.8 ml (38.1 mmol) of triethylamine and 1.6 ml (20.7 mmol) of methanesulfonyl chloride and the mixture was stirred at 0 °C for half an hour. Then 100 ml of methylene chloride and water were added to the reaction solution and the organic layer washed with water and then saturated citric acid aqueous solution, dried over the anhydrous sodium sulfate and concentrated under reduced pressure. The residue was refined by column chromatography on silica gel (eluted with ethyl acetate: hexane = 30:1) to give 1.3 g (64.6 % yield) of (2S,4R)-1-allyloxycarbonyl-2-[(Z)-3-methanesul fonylamino-1-propenyl]-4-methanesulfonyloxypyrrolidine. NMR(CDCl₃) δ :

1.95(m,1H), 2.50(m,1H), 2.93(s,3H), 3.03(s,3H), 3.62(m,2H), 4.00(m,2H), 4.44(d,2H), 4.80(q,1H), 5.05-5.41(m,4H), 5.70-5.95 (m, 2H)

REFERENCE EXAMPLE 4-6

To a solution of 1.3 g (3.4 mmol) of (2S,4R)-1-allyloxycarbonyl-2-[(Z)-3-methanesulfonylamino-1-propenyl]-4-methanesulfonyloxypyrrolidine obtained in

REFERENCE EXAMPLE 4-5 in 100 ml Of acetonitrile, 1.4 g

(12.26 mmol) of potassium thioacetate was added. The reaction solution was refluxed for 4 hours, cooled and concentrated under reduced pressure. Ethyl acetate and water were added to the residue. The organic layer was washed with water and then saturated sodium chloride aqueous solution, dried over the anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was refined by chromatography on silica gel

(eluted with ethyl acetate: hexane = 2:1) to give 730 mg

(60 % yield) of (2S,4S)-1-allyloxycarbonyl-2-[(Z)-3-methanesul f onylamino-1-propenyl ] -4-acetylthiopyrrolidine.

NMR(CDCl₃) δ :

1.60(m,1H), 2.30(S,3H), 2.60(m,1H), 2.95(S,3H) 3.10(m,1H), 3.95(m,3H), 4.60(m,3H), 5.10-5.50 (m, 3H), 5.62-5.95(m,3H)

REFERENCE EXAMPLE 4 -7

730 mg (2.01 mmol) of (2S,4S)-1-allyloxycarbonyl-2- [(Z)-3-methanesulfonylamino-1-propenyl]-4-acetylthio pyrrolidine obtained in REFERENCE EXAMPLE 4-6 was dissolved in 60 ml of methaneol and cooled to 0 °C. 2.3 ml of 2 N-sodium hydroxide was added to the mixture and the reaction solution was stirred for 3 minutes. Then, the mixture was immediately neutralized with saturated citric acid aqueous solution (pH = 3-4). The reaction mixture was concentrated under reduced pressure, and the residue was extracted with ethyl acetate. The organic layer was washed with water and the saturated sodium chloride aqueous solution, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was refined by column chromatography on silica gel (eluted with ethyl acetate:hexane=2:1) to give 610 mg (94.7 % yield) of (2S,4S)-1-allyloxycarbonyl-2-[(Z)-3-methanesulfonylamino-1-propenyl ]-4-mercaptopyrrolidine.

NMR(CDCl₃) δ :

1.63(m,1H), 1.70(d,1H), 2.60(m,1H), 2.98(s,3H) 3.23(m,1H), 3.60-4.05 (m, 3H), 5.54(m,3H), 5.13-5.58 (m,3H), 5.62-5.95(m, 3H)

REFERENCE EXAMPLE 5 (2S,4S)-1-allyloxycarbonyl-2-[(E)-3-methanesulfonyl amino-2-methyl-l-propenyl]-4-mercaptopyrrolidine

REFERENCE EXAMPLE 5-1 11.5 g (36.7 mmol) of (2S,4R)-1-allyloxycarbonyl-2- formyl-4-t-butyldimethylsilyloxypyrrolidine obtained by known processes and 26.5 g (73.4 mmol) of (carbetoxy ethylidene) triphenylphosphorane were dissolved in 250 ml of methylene chloride at room temperature and stirred for 20 hours. The mixture was concentrated under reduced pressure. The solvent mixture (1:4 v/v) of ethyl acetate and hexane was added to the mixture and the solid was removed by filtration. The filtrate was concentrated under reduced pressure and the residue was subject to column chromatography on silica gel (eluted with ethyl acetate: hexane = 1:7) to give 9.74 g (66.9 % yield) of ethyl 3-[(2S,4R)-1-allyloxycarbonyl-4-t-butyldimethylsilyloxypyrrolidin-2-yl]-2-methylacrylate.

NMR(CDCl₃) δ :

0.05(s,6H), 0.80(s,9H), 1.12(t,3H), 1.70(m,1H), 1.89(s,3H), 2.05(m,1H), 3.50(m,2H), 4.20(q,2H), 4.30-4.80(m,4H), 5.20(m,2H), 5.82(m,1H), 6.55(d,1H) REFERENCE EXAMPLE 5-2

The same procedure as in REFERENCE EXAMPLE 4-2 was carried out by using 9.74 g (24.5 mmol) of ethyl 3- [(2S,4R)-1-allyloxycarbonyl-4-t-buthyldimethylsilyloxy pyrrolidin-2-yl]-2-methylacrylate obtained in REFERENCE EXAMPLE 5-1 and 62 ml (62 mmol ) of 1M-diisobutyl aluminum hydride to obtain 8.15 g (93.7 % yield) of (2S,4R)-1-allyloxycarbonyl-2-[(E)-3-hydroxy-2-methyl-1-propenyl]-4-t-butyldimethylsilyloxypyrrolidine.

NMR(CDCl₃) δ :

0.05(S,6H), 1.81(s,9H), 1.63(m,1H), 1.70(s,3H), 2.03(m,1H), 3.50(m,2H), 4.01(s,2H), 4.01(s,2H), 4.30-4.70(m,4H), 5.20(m,3H), 5.90(m,1H)

REFERENCE EXAMPLE 5-3

The same procedure as in REFERENCE EXAMPLE 5-2 was carried out by using 8.15 g (22.96 mmol) of (2S,4R)-1- allyloxycarbonyl-2-[(E)-3-hydroxy-2-methyl-1-propenyl]- 4-t-butyldimethylsilyloxypyrrolidine obtained in REFERENCE EXAMPLE 5-2, 9.03 g (34.4 mmol) Of triphenyl phosphine, 5.74 g (39.01 mmol) of phthalimide and 5.4 ml (34.3 mmol) of diethyl azodicarboxylate to obtain 7.97 g (73.1 % yield) of (2S,4R)-1-allyloxycarbonyl-2-[(E)-3-phtalimido-2-methyl-1-propenyl]-4-t-butyldimethylsilyl oxypyrrolidine.

NMR(CDCl₃) δ: :

0.05(s,6H), 0.83(s,9H), 1.68(m,1H), 1.70(s,3H), 2.00(m,1H), 3.40(d,2H), 4.20-4.70 (m, 6H), 5.20(m,3H), 5.90(m,1H), 7.70-7.90(m,4H) REFERENCE EXAMPLE 5-4

The same procedure as in REFERENCE EXAMPLE 4-4 was carried out by using 7.97 g (16.96 mmol) of (2S,4R)-1-allyloxycarbonyl-2-[(E)-3-phthalimido-2-methyl-1-propenyl]-4-t-butyldimethylsilyloxypyrrolidine obtained in REFERENCE EXAMPLE 5-3, 3 ml (61.85 mmol) of hydrazine hydrate, 4.8 ml (62.01 mmol) of methanesulfonyl chloride and 12.1 ml (86.81 mmol) of triethylamine to obtain 4.53 g (61.7 % yield) of (2S,4R)-1-allyloxycarbonyl-2-[(E)-3-methanesulfonylamino-2-methyl-l-propenyl]-4-t-butyldimethylsilyloxypyrroiidine.

NMR(CDCl₃) δ : 0.05(s,6H), 0.80(S,9H), 1.65(m,1H), 1.70(s,3H),

2.03(m,1H), 2.90(S,3H), 3.40(m,2H), 3.60 (m, 2H) , 4.30- 4.70(m,4H), 5.10-5.30(m, 3H), 5.83(m,1H)

REFERENCE EXAMPLE 5-5

The same procedure as in REFERENCE EXAMPLE 4-5 was carried out by using 4.92 g (11.37 mmol) of (2S,4R)-1-allyloxycarbonyl-2-[(E)-3-methansulfonylamino-2-methyl-1-propenyl]-4-t-butyldimethylsilyloxypyrrolidine Obtained in REFERENCE EXAMPLE 5-4, 14.8 ml (14.8 mmol ) of lM-tetrabutyl ammonium fluoride, 3.6 ml (46.51 mmol) of methanesulfonyl chloride and 8.5 ml (60.98 mmol) of triethylamine to obtain 3.07 g (68.1 % yield) of ( 2 S , 4R ) -1-al lyloxycarbonyl -2- [ ( E ) -3 -methanesulfonylamino-2-methyl-1-propenyl]-4-methanesulfonyloxypyrrolidine.

NMR(CDCl₃) δ :

1.70(S,3H), 1.90(m,1H), 2.50(m,1H), 2.95(s,3H), 3.05(s,3H), 3.62(m,3H), 3.98(m,1H), 4.50-4.80 (m, 4H),

5.25(m,4H), 5.90(m,1H)

REFERENCE EXAMPLE 5-6 The same procedure as in REFERENCE EXAMPLE 4-6 was carried out by using 3.07 g (7.75 mmol) of (2S,4R)-1-allyloxycarbonyl-2-[(E)-3-methanesulfonylamino-2-methyl-1-propenyl]-4-methanesulfonyloxypyrrolidine obtained in REFERENCE EXAMPLE 5-5 and 2.91 g (25.6 mmol) Of potassium thioacetate to obtain 2.54 g (87.1 % yield) of

(2S,4S)-1-allyloxycarbonyl-2-[(E)-3-methanesulfonyl amino-2-methyl-1-propenyl]-4-acetylthiopyrrolidine.

NMR(CDCl₃) δ

1.62(m,1H), 1.80(s,3H), 2.30(s,3H), 2.54 (m, 1H), 2.95(s,3H), 3.23(m,1H), 3.62(m,2H), 4.10-3.85 (m, 2H), 4.50(m,4H), 5.15-5.42(m,3H), 5.90(m,1H) REFERENCE EXAMPLE 5-7

The same procedure as in REFERENCE EXAMPLE 4-7 was carried out by using 2.54 g (6.75 mmol) of (2S,4S)-1-allyloxycarbonyl-[(E)-3-methanesulfonylamino-2-methyl-1-propenyl]-4-acetylthiopyrrolidine obtained in REFERENCE EXAMPLE 5-6 and 8.1 ml of 2N-NaOH to obtain 1.89 g (83.1 % yield) of (2S,4S)-1-allyloxycarbonyl-2-[(E)-3-methanesulfonylamino-2-methyl-1-propenyl]-4-mercapto pyrrolidine.

NMR(CDCl₃) δ :

1.60(m,2H), 1.70(S,3H), 2.50(m,1H), 2.90(s,3H), 3.20(m,2H), 3.60(m,2H), 4.01(m,1H), 4.50(m,4H), 5.42-5.10(m,3H), 5.90(m,1H) REFERENCE EXAMPLE 6 ( 2S , 4S ) -1-allyloxycarbonyl-2- [ ( E ) -3- ( N , N-dimethylsul f amoyl amino ) -2 -methyl -1 -propenyl ] -4 -mercapto pyrrolidine

REFERENCE EXAMPLE 6-1

The same procedure as in REFERENCE EXAMPLE 5-4 was carried out by using 7.05 g (15 mmol) of (2S,4R)-1-allyioxycarbonyl-2-[(E)-3-phthalimido-2-methyl-1-propenyl]-4-t-butyldimetylsilyloxypyrrolidine obtained in REFERENCE EXAMPLE 5-3, 2.7 ml (55.66 mmol) Of hydrazine hydrate, 5.4 ml (38.74 mmol) of triethylamine and 2.8 ml (26.07 mmol) of dimethylsulfamoyl chloride to obtain 1.62 g (47 % yield) of (2S,4R)-1-allyloxy carbonyl-2-[(E)-3-(N,N-dimethylsulfamoylamino)-2-methyl- 1-propenyl]-4-t-butyldimethylsilyloxypyrrolidine.

NMR(CDCl₃) δ :

0.05(s,6H), 1.81(s,9H), 1.65(m,1H), 1.70(s,3H) 2.05(m,1H), 2.80(s,6H), 3.50(m,4H), 4.30-4.70 (m, 4H) , 5.20(m,3H), 5.90(m,1H)

REFERENCE EXAMPLE 6-2

The same procedure as in REFERENCE EXAMPLE 5-5 was carried out by using 1.62 g (3.49 mmol) of (2S,4R)-1-allyloxycarbonyl-2-[(E)-3-(N,N-dimethylsulfamoylamino)-2-methyl-1-propenyl]-4-t-butyldimethylsilyloxy pyrrolidine obtained in REFERENCE EXAMPLE 6-1, 4.3 ml

(4.3 mmol) of lM-tetrabutyl ammonium fluoride, 2.6 ml (18.65 mmol) of triethylamine and 1.1 ml (14.21 mmol) of methanesulfonyl chloride to obtain 1.33 g (89.0 % yield) of (2S,4R)-1-allyloxycarbonyl-2-[(E)-3-(N,N-dimethyl sulfamoylamino)-2-methyl-1-propenyl]-4-methanesul fonyloxypyrrolidine.

NMR(CDCl₃) δ : 1.70(s,3H), 1.90(m,1H), 2.50(m,1H), 2.80(s,6H),

3.04(s,3H), 3.50-4.01(m, 4H), 4.50-4.80 ( m, 3H),

5.20(m,4H), 5.90(m,1H)

REFERENCE EXAMPLE 6-3

The same procedure as in REFERENCE EXAMPLE 5-6 was carried out by using 1.33 g (3.11 mmol) of (2S,4R)-1-allyloxγcarbonyl-2-[(E)-3-(N,N-dimetylsulfamoylamino)-2-methyl-1-propenyl]-4-methanesulfonyloxypyrrolidine obtained in REFERENCE EXAMPLE 6-2 and 1.2 g (10.5 mmol) of potassium thioacetate to obtain 910 mg (71.7 % yield) of (2S, 4S)-1-allyloxycarbonyl-2-[ (E)-3-(N,N-dimethyisulfamoylamino)-2-methyl-1-propenyl]-4-acetylthiopyrrolidine.

NMR(CDCl₃) δ 1.60(m,1H), 1.75(S,3H), 2.30(s,3H), 2.58(m,1H), 2.80(s,6H), 3.23(dd,1H), 3.55(S,2H), 3.90(m,2H),

4.55(m,3H), 5.30(m,3H), 5.90(m,1H) REFERENCE EXAMPLE 6-4

The same procedure as in REFERENCE EXAMPLE 5-7 was carried out by using 910 mg (2.23 mmol) of (2S,4S)-1-allyloxycarbonyl-2-[(E)-3-(N,N-dimethylsulfamoylamino)-2-methyl-1-propenyl]-4-acetylthiopyrrolidine obtained in REFERENCE EXAMPLE 6-3 and 2.7 ml Of 2N-NaOH to Obtain 630 mg (77.2 % yield) of (2S,4S)-1-allyloxycarbonyl-2-[(E)-3-(N,N-dimethylsuIfamoylamino)-2-methyl-1-propenyl]-4-mercaptopyrrolidine.

NMR(CDCl₃) δ

1.60(s,2H), 1.70(s,3H), 2.52(m,1H), 2.80(s,6H), 3.10(m,1H), 3.50(s,2H), 4.00(m,1H), 4.50(m,3H), 5.30(m,3H), 5.90(m,1H)

REFERENCE EXAMPLE 7

(2S,4S)-1-allyloxycarbonyl-2-[(E)-3(4-morpholino sulfonylamino)-2-methyl-1-propenyl]-4-mercapto pyrrolidine REFERENCE EXAMPLE 7-1

To a solution of (2S,4R)-1-allyloxycarbonyl-2-[(E)- 3-phthalimido-1-propenyl]-4-t-butyldimethylsilyoxy pyrrolidine (3.53 g, 7.51 mmole) in anhydrous ethanol (80 ml) was added hydrazine hydrate (1.4 ml, 41.6 mmol).

The reaction mixture was stirred at room temperature overnight, filtered to remove insoluble material and the filtrate was concentrated under reduced pressure.

To the above residue was added ethyl acetate and the organic layer was stirred washed with 3N-ammonia solution and then saturated aqueous sodium chloride solution, and dried over anhydrous sodium sulfate and concentrated under reduced pressure. The concentrate was dissolved in dimethylformamide (50 ml) and triethylamine (1.3 ml, 94 mmol) and catechol sulfate (1.55 g, 9.01 mmol) was added successively thereto, and the reaction mixture was stirred for 1 hour. To the above solution was added ethyl acetate and water, and the ethyl actate layer was separated. The water layer was again extracted, a few times, with ethyl acetate. The combined organic layer was washed successively with saturated citric acid and then saturated aqueous sodium chloride, and dried over anhydrous sodium sulfate. The solvent was removed in vacuo, and the residue was subjected to silica gel column chromatography (eluting with ethyl acetate: hexane = 1:2) to give pure (2S,4R)-1-allyloxycarbonyl-2-[(E)-3-catecholsulfonylamino-2-methyl-1-propenyl]-4-t-butyldimethylsilyloxypyrrolidine (2.06 g, yield:52.2 %).

NMR(CDCl₃) δ

0.05(s,6H), 0.93(s, 9H), 1.70(s,3H), 2.01(m,1H), 3.43(m,2H), 3.70(d,2H), 4.35(m,1H), 4.60(m,3H), 5.30(m,3H), 5.90(m,1H), 6.80-7.30 (m, 4H )

REFERENCE EXAMPLE 7-2

To a solution of ( 2S , 4R )-1-allyloxycarbonyl-2- [ ( E ) - 3-catechol sul f onyl amino- 2 -methyl - 1 -propenyl ] - 4 -t- butyldimethylsilyloxypyrrolidine ( 2 . 06 g , 3 . 92 mmol ) in dioxane (60 ml) were added triethylamine (1.1 ml, 7.8 mmol) and morpholine (0.6 ml, 6.3 mmol). The reaction mixture was heated to reflux for 1.5 hour, and cooled to room temperature. The excess dioxane was removed in vacuo. To the residue was added ethyl acetate and the organic layer was washed with saturated aqueous sodium chloride, dried over anhydrous sodium sulfate and concentrated under reduced pressure. The crude product was purified by column chromatography on silica gel (eluting with ethyl acetate: hexane = 1:1 ) to give

(2S,4R)-1-allyloxycarbonyl-2-[(E)-(4-morpholinosulfonyl amino)-2-methyl-1-propenyl]-4-t-butyldimethylsilyloxy pyrrolidine (1.61 g, yield: 81.8 %). NMR(CDCl₃) δ:

0.05(S,6H), 0.93(S,9H), 1.75(s,3H), 2.01(m,1H), 3.10(m,4H), 3.46(d,2H), 3.60(d,2H), 3.70(m,4H), 4.30- 4.70(m,4H), 5.22(m,3H), 5.90(m,1H)

REFERENCE EXAMPLE 7-3

A mesylation of an amino moiety of (2S,4R)-1-allyloxycarbonyi-2-[(E)-(4-morphoiinosulfonylamino)-2-methyl-1-propenyl]-4-t-butyldimethylsilyloxypyrrolidine (1.61 g, 3.21 mmol) obtained in REFERENCE EXAMPLE 7-2 was conducted in the same manner as described in REFERENCE EXAMPLE 4-4 to REFERENCE EXAMPLE 4-7 to give (2S,4S)-1-allyloxycarbonyl-2-[(E)-3(4-morpholi nosulfonylamino)-2-methyl-1-propenyl]-4-mercaptopyrroli dine (0.8 g, yield: 61.6%).

NMR(CDCl₃) δ: 2.59(m,2H), 2.70(s,3H), 2.52(m,1H), 3.20(m,4H),

3.69(brs,2H), 3.70(m,4H), 4.01(m,1H), 4.50(m,4H), 5.15- 5 . 42 ( m, 3H ) , 5 . 90 ( m,1H)

REFERENCE EXAMPLE 8 (2S,4S)-1-allyloxycarbonyl-2-[(E)-2-chloro-3-methanesulfonylamino-1-propenyl]-4-mercaptopyrrolidine

REFERENCE EXAMPLE 8-1 To an ice cooled suspension of sodium hydride (2.09g of 60 % mineral oil, 52.27 mmol) in anhydrous tetrahydrofuran (135 ml) was added, successively and dropwise triethyl 2-chloro-2-phosphonoacetate (13.5 g, 52.27 mmole) and (2S, 4S)-1-allyloxycarbonyl-2-formyl-4-t-butyldimethylsilyloxypyrrolidine (13.65 g, 43.56 mmol) obtained by known processes. The mixture was stirred at the same temperature for 1 hour, and citric acid was added thereto to acidify the solution to pH 4.5. The excess organic solvent was evaporated in vacuo, and the resulting residue was diluted with methylene chloride, washed with water and then saturated aqueous sodium chloirde, dried over magenesium sulfate and concentrated under reduced pressure. The crude product was purified by flash chromatography on silica gel, eluting with a mixture of ethyl acetate: hexane (= 1:4) to give ethyl-3- [(2S,4R)-1-allyloxycarbonyl-4-t-butyldimethylsilyloxy pyrroiidin-2-yl]-2-chloroacrylate (10.4 g, yield: 57.1 %)

NMR(CDCl₃) δ:

0.05(s,6H), 0.80(S,9H), 1.30(t,3H), 1.80 (m, 1H), 2.20(m,1H), 3.50(m,2H), 4.05-4.40 (m, 3H), 4.59(m,2H), 4.90(m,1H), 5.20(m,2H), 5.90(m,1H), 7.01(t,1H) REFERENCE EXAMPLE 8-2 A crude product (8.7 g) obtained by the same process as that of REFERENCE EXAMPLE 1-2 from ethyl 3-[(2S,4R)-1-allyloxycarbonyl-4-t-butyldimethylsilyloxy pyrrolidin-2-yl]-2-chloloacrylate (6.0 g, 14.35 mmol) obtained in REFERENCE EXAMPLE 8-1 and diisobutylaluminum hydride (43 ml in 1.0 M toluene solution, 43 mmol) in anhydrous tetrahydrofuran (100 ml) was purified by column chromatography on silica gel (eluting with ethyl acetate: hexane = 1:2) to give (2S,4R)-1-allyloxycarbonyl-2-[(E)-chloro-3-hydroxy-1-propenyl]-4-t-butyldimethylsilyoxypyrroiidine (2.2g, yield: 40.8%) and (2S,4R)-1-allyloxycarbonyl-2-t-[(Z)-2-chloro-3-hydroxy-1-propenyl]-4-t-butyldimethylsilyloxypyrrolidine (0.8 g, yield:14.8 %).

(E) form

NMR(CDCl₃) δ: 0.05(s,6H), 0.80(s,9H), 1.70 (m, 1H), 2.20(m,1H),

3.50(m,2H), 4.10(S,2H), 4.30(m,1H), 4.55(m,2H), 4.80(q,1H), 5.20(m,2H), 5.90(m,2H)

(Z) form NMR(CDCl₃) δ

0.06(S,6H), 0.82(S,9H), 1.65(m,1H), 2.17(m,1H), 3.47(m,2H), 4.05(S,2H), 4.25(m,1H), 4.50(m,2H), 4.72(q,1H), 5.15(m,2H), 5.85(m,2H)

REFERENCE EXAMPLE 8-3

To an ice cooled solution of (2S,4R)-1-allyloxy carbonyl-2-[(E)-chloro-3-hydroxy-1-propenyl]-4-t- butyldimethylsilyloxypyrroiidine (0.8 g, 2.13 mmol) obtained in REFERENCE EXAMPLE 8-2 in anhydrous tetrahydrofuran (10 ml) was added dropwise triphenylphospine (0.84 g, 32.02 mmol) and phthalimide (0.55 g, 37.38 mmol), and diethyi azodicarboxylate (0.5 g, 31.75 mmole). The mixture was stirred for 1 hour, concentrated under reduced pressure to evaporate the organic solvent, and methylen chloride (200 ml) was added thereto. The organic layer was stirred, washed successively with water and saturated aqueous sodium chloirde, dried over anhydrous magnesium sulfate and concentrated in vacuo. The crude product was subjected to silica gel column chromatography (eluting with the mixture of .ethyl acetate: hexane = 1:3) to give (2S,4R)-1-allyloxycarbonyl-2-[(E)-chloro-3-phthalimido-1-propenyl]-4-t-butyldimethylsilyloxypyrrolidine (1.0 g, yield:92.6%).

NMR(CDCl₃) δ: 0.04(S,6H), 0.83(s,9H), 1.74(m,1H), 2.19(m,2H),

3.43(m,2H), 4.30-4.60(m,4H), 4.80(q,1H), 5.00-5.40(m,2H), 5.90(m,2H), 7.70-7.90 (m, 4H)

REFERENCE EXAMPLE 8-4

A crude product (0.8 g) obtained by the same process as that of REFERENCE EXAMPLE 1-4 from (2S,4R)-1-allyloxycarbonyl-2-[(E)-chloro-3-phthalimido-1-propenyl]-4-t-butyldimethylsilyloxypyrrolidine (1.0 g, 19.80 mmol), obtained in REFERENCE EXAMPLE 8-3, and ethanol (15 ml), hydrazine hydrate (0.27 ml, 55.3 mmol), triethylamine (0.7ml, 50.4 mmol), and methanesulfonyl chloride (0.215 ml, 7.72 mmole) was purified by column chromatography on silica gel (eluting with ethyl acetate :hexane = 1:1.5) to give (2S,4R)-1-allyloxycarbonyl-2- [(E)-chloro-3-methanesulfonylamino-1-propenyl]-4-t-butyl dimethylsilyloxypyrrolidme (0.61 g, yield: 82.4 %).

NMR(CDCl₃) δ: 0.03(s,6H), 0.83(s,9H), 1.70(m,1H), 2.18(m,1H),

3.00(m,2H) 3.43(m,2H), 4.30(m,1H), 4.50(m,2H), 4.80(q,1H), 4.90(m,2H), 5.20(m,2H), 5.90(m,2H)

REFERENCE EXAMPLE 8-5

(2S,4R)-1-allyloxycarbonyl-2-[(E)-chloro-3-methanesulfonylammo-1-propenyl]-4-t-butyldιmethylsιlyl oxypyrroiidme (0.61 g, 13.46 mmol), obtained in REFERENCE EXAMPLE 8-4, was dissolved m methanoi (50 ml), stirred at room temperature, and then HCl solution (1.2 ml) was added dropwise thereto. The mixture was stirred for 4 hour and concentrated under reduced pressure. The residual material was dissolved in methylene chloride, washed with water and then saturated aqueous sodium chloride, and the organic layer was dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated in vacuo. The resulting crude product was subjected to column chromatography on silica gel (eluting with chloroform: methanoi = 20:1) to afford (2S,4R)-1-allyloxycarbonyl-2-[(E)-chloro-3-methanesulfonylamino-1-propenyl]-4-hydroxypyrrolιdιne (0.36 g, yιeld:78.9 %).

NMR(CDCl₃) δ :

1.70(m,1H), 2.30(m,1H), 3.01(s,2H), 3.50(m,2H), 3.90(m,2H), 4.40-4.60(m, 2H), 4.80(q,1H), 5.20(m,2H), 5.90(m, 2H) REFERENCE EXAMPLE 8-6 To a solution of (2S,4R)-1-allyloxycarbonyl-2-[(E)-chloro-3-methanesulfonylamino-1-propenyl]-4-hydroxypyrrolidine (1.8 g, 5.32 mmol) obtained m REFERENCE EXAMPLE 8-5 in methylene chloride (60 ml) was added successively triethylamine (1.7 ml, 12.19 mmol) and methanesulfonyl chloride (0.49 ml, 6.33 mmol) at room temperature and the mixture was stirred for 40 minutes. The reaction mixture was washed successively with 1N- HC1, water, saturated sodium bicarbonate aqueous solution and finally distilled water, and dried over anhydrous magnesium sulfate, and then concentrated under reduced pressure to give (2S,4R)-1-allyloxycarbonyl-2-[(E)-chloro-3-methanesulfonylamιno-1-propenyl]-4-methanesulfonyloxypyrrolidine (2.1 g, yield: 95.0%), which was used at the next stage without purification.

NMR(CDCl₃) δ:

1.90(m,1H), 2.80(m,1H), 2.95(s,3H), 3.08(s,3H), 3.90(m,2H), 4.55(m,1H), 4.80(q,1H), 5.20(m,2H), 5.90(m,2H)

REFERENCE EXAMPLE 8-7 To a solution of (2S,4R)-1-allyloxycarbonyl-2-[(E)-chloro-3-methanesulfonylammo-1-propenyl]-4-methansulfonyloxypyrrolidine (2.1 g, 5.04 mmol) prepared m REFFERENCE EXAMPLE 8-6 in anhydrous acetonitrile (250 ml) was added potassium thioacetate (0.91 g, 7.95 mmol). The mixture was heated to reflux for 4 hours, cooled, and filtered to remove msoluble material. The filtrate was concentrated under reduced pressure. The residual material was dissolved m methylene chloride, washed with water and the organic layer was dried over anhydrous sodium sulfate, concentrated again under reduced pressure. The residue was purified by column chromatography on silica gel (eluting with chloroform : methanoi = 20:1) to give (2S,4S)-1-allyloxycarbonyl-2-[(E)-chloro-3-methanesulfonylamino-1-propenyl]-4-thioacetylpyrrolidine (1.4 g, yield: 67 %).

NMR(CDCl₃) δ:

1.70(m,1H), 2.32(s,3H), 2.80(m,1H), 3.00(s,3H), 3.30(m,1H), 3.90(m,2H), 4.50(m,2H), 4.80(q,1H), 5.20(m,2H), 5.90(m,2H)

REFERENCE EXAMPLE 8-8

To an ice cooled solution of (2S,4S)-1-allyloxycarbonyl-2-[(E)-chloro-3-methanesulfonylamino-1-propenyl]-4-thioacetylpyrrolidine (1.4 g, 3.53 mmol) obtained in REFERENCE EXAMPLE 8-7 in methanol (140 ml) was added 2N-NaOH (3.5 ml, 7.06 mmol) over the course of 1 minute and then an aqueous citric acid solution was added to adjust the pH of the solution 4.0. The mixture was concentrated under reduced pressure and diluted with methylene chloride. The organic layer was washed with water, dried over anhydrous magnesium sulfate, concentrated in vacuo to give (2S,4S)-1-allyloxycarbonyl-2-[(E)-chloro-3-methanesulfonylamino-1-propenyl]-4-mercaptopyrrolidine (1.16 g, yield: 92.8 %), which was used at the next stage without purification.

NMR(CDCl ₃) δ:

1.69(m,1H), 2.79(m,1H), 3.03(s,3H), 3.25(m,1H), 3.91(m,2H), 4.49(m,2H), 4.82(q,1H), 5.19(m,1H), 5.89(m, 2H) REFERENCE EXAMPLE 9 (2S,4R)-1-allyloxycarbonyl-2-[ (E)-3-methanesulfonyl-1-propenyl]-4-t-butyldimethylsilyloxy pyrrolidine REFERENCE EXAMPLE 9-1

To an ice cooled solution of 2-aminoethanol 5 g (81.9 mmol) in dichloromethane (200 ml) was added dropwise triethylamine (28.5 ml, 204.8 mmol) and methanesulfonyl chloride (14.6 ml, 188.4 mmol). The mixture was stirred at 0 °C for 30 minutes, and additional dichloromethane (100 ml) was added thereto. The organic layer was washed with a small amount of diluted-HCl solutions and then saturated aqueous sodium chloride, dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure to give N, O-dimethanesulfonylethanoiamine (14.5 g, yield: 81.5 %). NMR(CDCl₃) δ:

3.02(s,3H), 3.09(s,3H), 3.51(q,2H), 4.35(t,3H), 5.10(bs,1H) REFERENCE EXAMPLE 9-2

A solution of N,O-dimethanesulfonylethanolamine

(8.3 g, 38.2 mmol) obtained in REFERENCE EXAMPLE 9-1 and lithium bromide (4.0 g, 46.1 mmol) in acetone (120 ml) was heated to relux for 2 hours, and then chloroform (50 ml) was added thereto. The resulting precipitate was filtered off and the filtrate was concentrated in vacuo. The crude product obtained was purified by silica gel chromatography (eluting with mixture of chloroform: acetone = 10:1) to give N-methanesulfonyl-2-bromoethylamine (8.3 g, yield: 89 %).

NMR(CDCl₃) δ:

3.03(s,3H), 3.54-3.57(m,4H), 4.90(bs,1H)

REFERENCE EXAMPLE 9-3

N-methanesulfonyl-2-bromoethylamine (8.0 g, 39.6 mmol) obtained in REFERENCE EXAMPLE 9-2 and triphenyl phophine (11.5 g, 43.8 mmol) were dissolved in toluene (80 ml). The solution was heated to reflux for 9 hours, cooled to room temperature and concentrated under reduced pressure. The crude material was dissolved in dichloromethane and ether to crystallize the desired product. The solid which formed was obtained by filtration and dried under vacuum to give [(2-methanesulfonylamino)ethyl]triphenylphosphonium bromide (11.5 g, yield:62.5 %). NMR(DMSO-d₆) δ:

2.93(s,3H), 3.29(m,2H), 3.85(m,2H), 7.50(t,1H), 7 . 70-8 . 01 ( m, 15H )

REFERENCE EXAMPLE 9-4 To a dry-ice (-78 °C) cooled solution of [(2-methanesulfonylamino)ethyl]triphenylphosphonium bromide (0.39 g, 0.84 mmol) obtained in REFERENCE EXAMPLE 9-3 in tetrahydrofuran (4 ml) was added dropwise and carefully n-butyllithium (1.26 ml of hexane solution, 1.26 mmol) solution. The reaction solution was warmed to -40 °C, stirred for 10 minutes, and cooled to -78 °C again. Hexamethylphosphorous triamide (0.37 ml, 2.13 mmol) was added thereto. The mixture was stirred for 10 minutes, and to the above solution was added a solution of (2S,4R)-1-allyloxycarbonyl-2-formyl-4-t-butyldimethyl silyloxypyrrolidine (0.131 g, 0.418 mmol), obtained by known processes, in tetrahydrofuran (4 ml). The reaction mixture was warmed slowly to room temperature, concentrated under reduced pressure, and the residue was dissolved in ethyl acetate, washed with water, dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated in vacuo. The concentrate was subjected to column chromatography on silica gel (eluting with ethyl acetate: hexane = 1:2) to give (2S,4R)-1-allyloxycarbonyl-2-[(E)-3-methanesul fonylamino-1-propenyl]-4-t-butyldimethylsilyloxy pyrrolidine (0.106 g, yield: 60.4 %). NMR(CDCl₃) δ:

0.03(s,6H), 0.85(s,9H), 1.68-1.78 (m, 1H), 1.98- 2.06(m,1H), 2.93(s,3H), 3.45-3.47 (d, 2H), 3.70-3.74(m,2H), 4.31-4.43 (m, 2H), 4.51-4.54 (m, 3H), 5.14-5.30(m,2H), 5.62(bs,2H), 5.81-5.98 (m, 1H)

Detailed Description of the Preferred Embodiments EXAMPLE 1

(1R,5S,6S)-2-[(2S,4S)-2-[ (E)-3-methanesulfonyl amino-1-propenyl]pyrrolidin-4-ylthiol-6-[ (R)-1-hydroxyethyl]-1-methyl-1-carbapen-2-em-3-carboxylic acid

EXAMPLE 1-1

To an ice-cooled solution of allyl (1R,5S,6S)-2-diphenoxyphosphoryioxy-6-[(R)-1-hydroxyethyl]-1-methyl- 1-carbapen-2-em-3-carboxylate (4.12 g, 9.17 mmol) obtained by known processes, in anhydrous acetonitrile (50 ml) was added diisopropylethylamine (1.6 ml, 9.18 mmol). The above solution was stirred for 5 minutes. And then the solution of (2S,4S)-1-allyloxycarbonyl-2- [ ( E ) -3-methanesulfonγlamino-1-propenγl ]-4- mercaptopyrrolidine (2.45 g, 7.01 mmol) obtained in REFERRENCE EXAMPLE 1-7 in anhydrous acetonitrile (10 ml) was added dropwise thereto. The reaction mixture was stirred for 4 hours and concentrated under reduced pressure. The residue was diluted with ethylacetate, washed with distilled water, dried over anhydrous magnesium sulfate and the solvent was removed in vacuo. The residue was subjected to column chromatography on silica gel (eluting with ethylacetate: hexane = 2:1, and chloroform: methanoi = 50:1) to give pure allyl (1R,5S, 6S)-2-[(2S,4S)-1-allyloxycarbonyl-2-[(E)-3-methanesul fonylamino-1-propenyl]pyrrolidin-4-ylthio]-6-[(R)-1-hydroxyethyl]-1-methyl-1-carbapen-2-em-3-carboxylate (2.8 g, yield: 65 %).

NMR(CDCl₃) δ

1.19-1.24(d, 3H), 1.30-1.33 (d, 3H), 1.78-1.84 (m, 1H), 2.52-2.59(m, 1H), 2.43(s, 3H), 3.20-3.42 (m, 3H), 3.70-3.75(m, 3H), 3.84-4.05(m, 1H), 4.41-4.52 (m, 1H), 4.53-4.55(m, 2H), 4.61-4.70(m, 1H), 4.77-4.81 (m, 1H), 5.17-5.30(m, 3H), 5.37-5.47 (m, 2H), 5.55-5.68 (m, 2H), 5.84-5.95(m, 2H)

EXAMPLE 1-2 To an ice cooled mixture of ally (1R,5S,6S)-2- [(2S,4S)-1-allyloxycarbonyl-2-[(E)-3-methanesulfonyl amino-1-propenyl]pyrrolidin-4-ylthio]-6-[(R)-1-hydroxy ethyl]-1-methyl-1-carbapen-2-em-3-carboxylate (2.8 g, 4.91 mmol), obtained in EXAMPLE 1-1, and distilled water (0.4 ml, 22.2 mmol) in anhydrous dichloromethane (59 ml) was added successively bis (triphenylphosphine) palladium(II) chloride (0.03 g, 0.043 mmol) and tributyltin hydride (2.9 ml, 10.78 mmol). The solution was stirred for 10 minutes, extracted with distilled water (100 ml × 3), washed with chloroform (100 ml × 2), and the aqueous layer was freeze-dried to give (1R,5S, 6S)-2-[(2S,4S)-2-[(E)-3-methanesulfonylamino-1-propenyl]-pyrrolidin-4-ylthio]-6-[(R)-1-hydroxyethyl]-1-methyl-1-carbapen-2-em-3-carboxylic acid (1.31 g, yield : 60 % ) . NMR(DMSO-d₆ + D₂O)δ:

1.02-1.05(d, 3H), 1.10-1.13(d,3H), 1.45-1.63 (m, 1H), 2.41-2.50(m,1H), 2.86(S,3H), 3.01-3.12 (m, 3H), 3.45- 3.57(m,3H), 3.87-4.05(m,.2H), 5.77-5.88 (m, 2H)

EXAMPLE

(1R,5S.6S)-2-[(2S.4S)-2-[(E)-3-methanesulfonyl amino-1-methylpropenyl1-pyrrolidin-4-ylthio1-6-[(R)-1- hydroxyethyl]-1-methyl-1-carbapen-2-em-3-carboxylic acid

EXAMPLE 2-1 To an ice cooled solution of allyl (1R,5S,6S)-2-diphenoxyphosphoryloxy-6-[(R)-1-hydroxyethyl]-1-methyl- 1-carbapen-2-em-3-carboxylate (0.5 g, 1.0 mmol), obtained in known processes, and diisopropylethylamine (0.21 ml, 1.2mmol) in acetonitrile (10 ml) was added dropwise a solution of (2S,4S)-1-allyloxycarbonyl-2- [ ( E )-3-methanesulfonylamino-1-propenyl ]-4-mercaptopyrrolidine (0.33 g, 1.0 mmol), obtained in

REFERRENCE EXAMPLE 2-6 in acetonitrile (5 ml). The above reaction solution was stirred for 3 hours and concentrated under reduced pressure. The crude product was purified by column chromatography on silica gel

(eluting with chloroform: acetone = 3:1) to give allyl

(1R,5S,6S)-2-[ (2S,4S)-1-allyloxycarbonyl-2-[(E)-3-methanesulfonylamino-1-methyl-propenyl]pyrroiidin-4-ylthio]-6-[(R)-1-hydroxyethyl]-1-methyl-1-carbapen-2-em- 3-carboxylate (0.16 g, yield: 28 %).

NMR ( CDCl₃) δ:

1.22(d,3H), 1.34(d,3H), 1.64(s,3H), 1.84(m,1H), 2.51(m,1H), 2.94(s,3H), 3.30(m,2H), 3.61(m,1H),

3.81(m,2H), 4.21(m,2H), 4.52(m,2H), 5.19-5.50 (m, 5H), 5.90(m,2H)

EXAMPLE 2-2 To a solution of allyl (IR, 5S, 6S)-2-[ (2S, 4S)-1- allyloxycarbonyl-2-[(E)-3-methanesulfonylamino-1-methyl- propenyl ]pyrrolidin-4-ylthio ]-6- [ (R)-1-hydroxyethyl ]-1- methyl-1-carbapen-2-em-3-carboxylate (0.16 g, 0.28 mmol) obtained in EXAMPLE 2-1, and distilled water (0.22 ml) in dichloromethane (5.5 ml) was added of bis(triphenyl phosphine) palladium (II) chloride (10 mg, 0.014 mmol) and tributyltin hydride (0.15 ml, 0.55 mmol). The solution -was stirred for 20 minutes, and then extracted with distilled water (10 ml x 3). The aqueous layer was washed with chloroform and freeze-dried to give (1R,5S,6S)-2-[(2S,4S)-2-[(E)-3-methanesulfonylamino-1-methylpropenyl]-pyrrolidin-4-ylthio]-6-[(R)-1-hydroxyethyl]-1-methyl-1-carbapen-2-em-3-carboxylic acid (55 mg, yield: 43 %).

NMR(D₂O) δ:

1.08(d,3H), 1.15(d,3H), 1.68(S,3H), 1.82(m,1H), 2.60(m,1H), 2.95(S,3H), 3.22(m,3H), 3.54(m,1H), 3.68(d,2H), 3.90(m,1H), 4.00-4.21 (m, 3H), 5.60(t,1H)

EXAMPLE 3 (1R,5S,6S)-2-[(2S,4S)-2-[(E)-3-fluoro-1-propenyl] pyrroiidin-4-ylthio]-6-[(R)-1-hydroxyethyl]-1-methyl-1- carbapen-2-em-3-carboxylic acii EXAMPLE 3-1

Allyl (1R,5S,6S)-2-diphenoxyphosphoryloxy-6-[(R)-1-hydroxyethyl]-1-methyl-1-carbapen-2-em-3-carboxylate (0.23 g, 0.5 mmol), obtained by known processes, and (2S,4S)-1-allyloxycarbonyl-2-[(E)-3-fluoro-1-propenyl]-4-mercaptopyrrolidine (0.12 g, 0.5 mmol), obtained in REFERENCE EXAMPLE 3-4, were treated by the same process as that of EXAMPLE 1-1 to give crude material (87 mg), which was subjected to silica gel chromatography (eluting with chloroform: acetone = 4:1) to give allyl (1R,5S,6S)-2-[(2S,4S)-1-allyloxycarbonyl-2-[(E)-3-fluoro-1-propenyl]pyrrolidin-4-ylthio]-6-[(R)-1-hydroxy ethyl]-1-methyl-1-carbapen-2-em-3-carboxylate (24 mg, yield: 11 %).

NMR(CDCl₃) δ:

1.23(d,3H), 1.38(d,3H), 1.75(m,1H), 2.57(m,1H), 3.10(dd,1H), 4.20(m,2H), 4.95(d,1H), 5.10-5.50 (m, 5H), 5.80(m,4H)

EXAMPLE 3-2 Allyl (1R,5S,6S)-2-[(2S,4S)-1-allyloxycarbonyl-2- [(E)-3-fluoro-1-propenyl]pyrrolidin-4-ylthio]-6-[(R)-1- hydroxyethyl]-1-methyl-1-carbapen-2-em-3-carboxylate (22 mg, 0.05 mmol), obtained in EXAMPLE 3-1, was treated by the same process as that of EXAMPLE 1-2 to give (1R,5S, 6S)-2-[(2S,4S)-2-[(E)-3-fluoro-1-propenyl]pyrrolidin-4-ylthio]-6-[(R)-1-hydroxyethyl]-1-methyl-1-carbapen-2-em-3-carboxylic acid (12 mg, yield: 73 %).

NMR(D₂O) δ :

1.07(d,3H), 1.16(d,3H), 2.50(m,2H), 3.20(m,1H), 3.30(dd,1H), 3.43-3.70(m,3H), 3.83(m,1H), 3.92-4.13(m,2H), 5.05(d,1H), 5.10(d,1H), 5.62(m,1H)

EXAMPLE 4 (1R,5S,6S)-2-[(2S,4S)-2-[(Z)-3-methanesulfonyl amino-1-propenyl]pyrrolidin-4-ylthio]-6-[(R)-1-hydroxy ethyl]-1-methyl-1-carbapen-2-em-3-carboxylic acid

EXAMPLE 4-1

To a solution of allyl (1R,5S,6S)-2-diphenoxyphos phoryloxy-6-[(R)-1-hydroxyethyl]-1-methyl-1-carbapen-2- em-3-carboxylate (980 mg, 1.96 mmol), obtained by known processes, and (2S,4S)-allyloxycarbonyl-2-[(Z)-3-methane sulfonylamino-1-propenyl]-4-mercaptopyrrolidine (640mg, 2.0 mmol), obtained in REFERENCE EXAMPLE 4-7, in anhydrous acetonitrile (60 ml) was added N,N- diisopropylethylamine (0.37 ml, 2.12 mol). The above mixture was stirred for 6 hours and concentrated under reduced pressure at room temperature. Ethyl acetate was added thereto and the organic layer was washed with water and then saturated aqueous sodium chloride, dried over anhydrous sodium sulfate and concentrated in vacuo. This crude product was purified by column chromatography on silica gel (eluting with chloroform: methanoi = 30:1) to give pure allyl (1R,5S,6S)-2-[(2S,4S)-1-allyloxycarbonyl-2-[(Z)-3-methanesulfonylamino-1-propenyl]pyrrolidin-4-ylthio]-6-[(R)-1-hydroxyethyl]-1-methyl-1-carbapen-2-em-3-carboxylate (560 mg, yield: 49.0 %).

NMR(CDCl₃) δ:

1.25(d,3H), 1.33(d,3H), 1.70(m,1H), 2.60(m,1H), 2.95(s,3H), 3.30(m,3H), 3.63(m,2H), 3.95(m,2H), 4.20(m,2H), 4.52(d,2H), 4.60-4.83 (m,3H), 5.20-6.02 (m, 8H) EXAMPLE 4-2

To an ice cooled solution of allyl (1R,5S,6S)-2- [(2S,4S)-1-allyloxycarbonyl-2-[(Z)-3-methanesulfonyl amino-1-propenyl]pyrrolidin-4-ylthio]-6-[(R)-1-hydroxyethyl]-1-methyl-1-carbapen-2-em-3-carboxylate (1.58 g, 2.27 mmol) in methylenechloride (54 ml) were added successively distilled water (0.196 ml), bis(tripenyiphosphine)palladium (II) chloride (90 mg, 0.14 mmol) and tributyltin hydride (1.39 ml, 5.18 mmol). The above solution was stirred for 20 minutes at 0°C, extracted with distilled water and the aqueous layer was washed with chloroform several times, and filtered through 0.5 μm filter paper. The trace chloroform in the aqueous layer was removed thoroughly under reduced pressure and the aqueous residue was freezedried to give (1R,5S,6S)-2-[(2S,4S)-2~[(Z)-3-methanesulfonylamino-1-propenyl]pyrrolidin-4-ylthio]-6-[(R)-1-hydroxyethyl]-1-methyl-1-carbapen-2-em-3-carboxylic acid (1.1 g, yield: 89 %).

NMR (DMSO-d₆) δ:

1.05(m,6H), 1.55(m,1H), 2.50(m,1H), 2.90- 3.30(m,4H), 3.50-3.80(m,4H),3.90 (s, 3H), 4.01 (m, 1H), 4.05(m,1H), 4.30(m,1H), 5.70(m,2H), 7.10(brs.1H) EXAMPLE 5

(1R,5S,6S)-2-[(2S,4S)-2-[(E)-3-methanesulfonyl amino-2-methyl-1-propenyl]pyrrolidin-4-ylthio1-6-[(R)-1- hydroxyethyl]-1-methyl-1-carbapen-2-em-3-carboxylic acid.

EXAMPLE 5-1 Allyl (1R,5S,6S)-2-[(2S,4S)-1-allyloxycarbonyl-2- [(E)-3-methanesulfonylamino-2-methyl-1-propenyl]pyrrolidin-4-ylthio]-6-[(R)-1-hydroxyethyl]-1-methyl-1-carbapen-2-em-3-carboxylate (1.58 g, yield: 47.8 %) was obtained from allyl (1R,5S,6S)-2-diphenoxyphosphoryloxy- 6-[(R)-1-hydroxyethyl]-1-methyl-1-carbapen-2-em-3-carboxylate (2.97 g, 5.94 mmol) obtained by known processes, and (2S,4S)-1-allyloxycarbonyl-2-[(E)-3-methanesulfonylamino-2-methyl-1-propenyl]-4-mercapto pyrrolidine (1.89 g, 5.66 mmol) obtained in REFERENCE

EXAMPLE 5-7 by the same process as that of EXAMPLE 4-1.

NMR(CDCl₃) δ 1.25(d,3H), 1.30(d,3H), 1.65 (m, 1H), 1.70(s,3H),

2.60(m,2H), 1.90(s,3H), 3.30(m,3H), 3.60 (m, 3H), 3.98(m,1H), 4.20(m,2H), 4.50(d,2H), 4.58-4.82 (m, 3H), 5.20-5.50(m,5H), 5.90(m,2H) EXAMPLE 5-2

(1R,5S,6S)-2-[(2S,4S)-2-(E)-3-methanesulfonyl amino-2-methyl-1-propenyl]pyrroiidin-4-ylthio]-6-[(R)-1-hydroxyethyl]-1-methyl-1-carbapen-2-em-3-carboxylic acid (1.1 g, yield: 89 %) was obtained from allyl (1R,5S,6S)- 2-[(2S,4S)-1-allyloxycarbonyl-2-[(E)-3-methanesulfonyl amino-2-methyl-1-propenyl]pyrrolidin-4-ylthio]-6-[(R)-1- hydroxyethyl]-1-methyl-1-carbapen-2-em-3-carboxylate (1.58 g, 2.71 mmol) obtained in EXAMPLE 5-1, bis (triphenylphospine) palladium (II) chloride (90 mg, 0.14 mmol), and tributyltinhydride (1.39 ml, 5.18 mmol) by the same process as that of EXAMPLE 4-2.

NMR(DMSO-d₆) δ:

1.10(m,6H), 1.50(m,1H), 1.70(s,3H), 2.40ms,1H), 2.87(s,3H), 3.01-3.30(m,4H), 3.50(s,3H), 3.70-4.10 (m,3H), 5.48(d,1H), 7.23(t.1H)

EXAMPLE 6 (1R,5S,6S)-2-[(2S,4S)-2-[(E)-3-(N,N-dimethylsul famoylamino-2-methyl-1-propenyl]pyrrolidin-4-ylthio]-6- [ (R)-1-hydroxyethyl]-1-methyl-1-carbapen-2-em-3-carboxylic acid EXAMPLE 6-1

Allyl (1R,5S,6S)-2-[(2S,4S)-1-allyloxycarbonyl-2- [ (E)-3-(N,N-dimethylsuIfamoylamino)-2-methyl-1- propenyl]pyrrolidin-4-ylthio]-6-[(R)-1-hydroxyethyl]-1- methyl-1-carbapen-2-em-3-carboxylate (360 mg, yield:

34.1 %) was obtained from allyl (1R,5S,6S)-2- diphenoxyphosphoryloxy-6-[(R)-1-hydroxyethyl]-1-methyl- 1-carbapen-2-em-3-carboxylate (630 mg, 1.72 mmol) obtained by known processes, (2S, 4S)-1-aliyloxycarbonyl-2-[(E)-3-(N,N-dimethylsulfamoylamino)-2-methyl-1-propenyl]-4-mercaptopyrrolidine (902 mg, 1.81 mmol) obtained in REFERENCE EXAMPLE 6-4, and N,N-diisopropyl ethylamine (0.33 ml, 1.89 mmol) by the same process as that of EXAMPLE 4-1.

NMR(CDCl₃) δ:

1.25(d,3H), 1.30(d,3H), 1.70(m,1H), 1.72(s,3H),

2.50(m,2H), 2.80(s,6H), 3.30(m,3H), 3.58(d,2H),

4.00(m,1H), 4.22(m,2H), 4.50-4.87 (m, 3H), 5.19-5.45 (m,5H), 5.90(m,2H)

EXAMPLE 6-2

(1R,5S,6S)-2-[(2S,4S)-2-[(E)-3-(N,N-dimethylsul famoylamino-2-methyl-1-propenyl-4-ylthio]-6-[(R)-1-hydroxyethyl]-1-methyl-1-carbapen-2-em-3-carboxylic acid (164 mg, 56.9 mmol) was obtained from allyl (1R,5S,6S)- 2-[(2S,4S)-1-allyloxycarbonyl-2-[(E)-3-(N-N-dimethylsulfamoylamino-2-methyl-1-propenyl]pyrrolidin-4-ylthio]-6- [(R)-1-hydroxyethyl]-1-methyl-1-carbapen-2-em-3-carboxylate (360 mg, 0.59 mmol) obtained in EXAMPLE 6-1, distilled water (0.043 ml), bis (triphenylphosphine) palladium(II) chloride (20 mg, 0.03 mmol), and tributyltin hydride (0.303 ml, 1.13 mmol) by the same process as that of EXAMPLE 4-2.

NMR(CDCl₃) δ:

1.07(d,3H), 1.15(d,3H), 1.40(m,1H), 1.62(s,3H), 2.40(m,1H), 2.80(s,6H), 2.95-3.50 (m, 4H), 3.70-4.10 (m,4H), 5.50(d,1H), 7.40(t,1H) EXAMPLE 7 (1R,5S,6S)-2-[(2S,4S)-2-[(E)-3-(4-morpholinosul fonylamino)-2-methyl-1-propenyl]pyrrolidin-4-ylthio]-6- [ (R)-1-hydroxyethyl]-1-methyl-1-carbapen-2-em-3-carboxylic acid

EXAMPLE 7-1

Allyl (1R,5S,6S)-2-[(2S,4S)-1-allyloxycarbonyl-2- [ (E)-3-(4-morpholinosulfonylamino)-2-methyl-1- propenyl]pyrrolidin-4-ylthio]-6-[(R)-1-hydroxyethyl]-1- methyl-1-carbapen-2-em-3-carboxylate 583 mg (yield: 45%) was obtained from allyl (1R,5S,6S)-2-diphenoxyphos phoryloxy-6-[(R)-1-hydroxyethyl]-1-methyl-1-carbapen-2- em-3-carboxylate (1.04 g, 2.08 mmol) obtained by well known processes, (2S,4S)-1-allyloxycarbonyl-2-[(E)-3- (4-morpholinosulfonylamino)-2-methyl-1-propenyl]-4- mercaptopyrrolidine (800 mg, 1.98 mmol) obtained in REFERENCE EXAMPLE 7-3, and N,N-diisopropylethylamine (0.38 ml, 2.18 mmol) by the same process as that of EXAMPLE 4-1.

NMR(CDCl₃) δ:

1.24(d,3H), 1.34(d,3H), 1.69(m,1H), 1.70(s,3H), 2.50(m,1H), 3.20(m,7H), 3.58(d,2H), 3.75(m,4H), 3.99(m,1H), 4.20(m,2H), 4.40-4.80 (m, 5H), 5.20-5.40 (m,5H), 5.89(m,2H)

EXAMPLE 7-2 (1R,5S,6S)-2-[(2S,4S)-2-[(E)-3-(4-morpholinosulfonγlamino)-2-methyl-1-propenyl]pyrrolidin-4-ylthio]-6- [ (R)-1-hydroxyethyl]-1-methyl-1-carbapen-2-em-3-carboxylic acid (354 mg, yield: 75 %) was obtained from allyl (1R,5S,6S)-2-[(2S,4S)-1-allyloxycarbonyl-2-[(E)-3- (4-molpolynosulfonylamino)-2-methyl-1-propenyl] pyrrolidin-4-ylthio]-6-[(R)-1-hydroxyethyl]-1-methyl-1- carbapen-2-em-3-carboxylate (580 mg, 0.89 mmol) obtained in EXAMPLE 7-1, distilled water (0.065ml), bis

(triphenylphosphine) palladium(II) chloride 30 ml (0.045 mmol), and tributyltin hydride (0.46 ml, 1.71 mmol) by the same process as that of EXAMPLE 4-2. NMR(DMSO-d₆) δ:

1.06(d,3H), 1.14(d,3H), 1.40(m,1H), 1.61(S,3H), 2.40(m,1H), 2.90-3.50(m,8H), 3.69(m,4H), 3.7-4.09 (m,4H), 5.4(d,1H), 7.39(t,1H)

EXAMPLE 8 (1R,5S,6S)-2-[(2S,4S)-2-[(E)-2-chloro-3-methanesul fonylamino)-1-methyl-1-propenyl]pyrrolidin-4-ylthio]-6-[(R)-1-hydroxyethyl]-1-methyl-1-carbapen-2-em-3-carboxylic acid

EXAMPLE 8-1

Allyl (1R,5S,6S)-2-[(2S,4S)-1-allyloxycarbonyl-2- [(E)-2-chloro-3-methanesulfonylamino-1-propenyl] pyrrolidin-4-ylthio]-6-[(R)-1-hydroxyethyl]-1-methyl-1-carbapen-2-em-3-carboxylate (0.706 g, yield: 42 %) was obtained from allyl (1R,5S,6S)-2-diphenoxyphosphoryloxy- 6-[(R)-1-hydroxyethyl]-1-methyl-1-carbapen-2-em-3-carboxylate (1.25 g, 2.78 mmol) obtained by well known processes, (2S,4S)-1-allyloxycarbonyl-2-[(E)-2-chloro-3-methanesulfonylamino)-1-propenyl]-4-mercaptopyrroiidine (0.96g, 2.7mmθl) obtained in REFERENCE EXAMPLE 8-8, and

N,N-diisopropylethylamine (0.28 ml, 2.9 mmol) by the same process as that of REFERENCE EXAMPLE 4-1. NMR(CDCl₃) δ:

1.21(d,3H), 1.30(d,3H), 1.70(m,1H), 2.59(m,1H), 2.90(S,3H), 3.30(m,3H), 3.60(m,3H), 4.20(m,2H), 4.45- 4.80(m,5H), 5.10-5.45(m,5H), 5.90(m,2H)

EXAMPLE 8-2

(1R,5S,6S)-2-[(2S,4S)-2-[(E)-2-chloro-3-methanesul fonylamino)-1-propenyl]pyrrolidin-4-ylthio]-6-[(R)-1-hydroxyethyl]-1-methyl-1-carbapen-2-em-3-carboxylic acid (0.36 g, yield: 64.7 %) was obtained from allyl (1R,5S,6S)-2-[(2S,4S)-1-allyloxycarbonyl-2-[(E)-2-chloro-3-methane sulfonylamino)-1-propenyl]pyrrolidin-4-ylthio]-6-[(R)-1-hydroxyethyl]-1-methyl-1-carbapen-2-em-3-carboxylate (0.7 g, 1.16 mmol) obtained in EXAMPLE 8-1, bis (triphenylphosphine) palladium(II) chloride (42 mg, 0.06 mmol), and tributyltin hydride (0.73 ml, 2.32 mmol) by the same process as that of EXAMPLE 4-2.

NMR(DMSO-d₆) δ:

1.05(m,6H), 1.50(m,1H), 2.40(m,1H), 2.84(s,3H), 3.02-3.40(m,4H), 3.80-4.10 (m, 3H), 5.59(d,1H), 7.30(t,1H)

Claims

WHAT I S CLAIMED I S :

1 . A compound of formula ( I )

or a pharmaceutically acceptable salt or ester thereof; wherein R₁ is selected from the group consisting of a hydrogen atom or a methyl group, R₂ is selected from the group consisting of a metal or a nonmetal salt group, or a carboxy protecting group, or a negative charge, R₃ is selected from the group consisting of a hydrogen atom or an imino protecting group, or a pharmaceutically acceptable salt and R₄ and R₅ are each selected from the group consisting of a hydrogen atom, a hydroxy group, a cyano group, a halogen atom, methanesulfonyl group, N-methylsulfamoyl group, N,N-dimethylsulfamoyl group, or a lower alkyl group which may be optionally substituted with appropriate substituents, or a carbamoyl group which may be optionally substituted with appropriate substituents. A is selected from the group consisting of a halogen atom or the group of the following formula (1) or (2);

(wherein R₆ and R₇ are each selected from the group consisting of hydrogen atom or a lower alkanesulfonyl group, a N-(lower)alkylsulfamoyl group, or a N,N- (lower)dialkyl sulfamoyl group, wherein one of R₆ and R₇ is hydrogen, the other is other nonhydrogen group.)

(wherein R₆ is the same as defined above and R₈ is selected from the group consisting of a carbamoyl group which may be optionally substituted with appropriate substituents, or heterocyclic group of 5- to 6- membered ring consisting of 1 to 4 heteroatoms such as a nitrogen atom, an oxygen atom or a sulfur atom which may be optionally substituted with appropriate substituents), and n is 0 to 3.

2. The compound of formula (I) according to claim 1, wherein R₁ is a methyl group, and R₂ is a hydrogen atom

3. The compound of formula (I) according to claim 1, wherein each of R₁ and R₂ is a hydrogen atom.

4. The compound of formula (I) according to claim 1, wherein R₁ is a methyl group, and each of R₂ and R₃ is a hydrogen.

5. The compound of formula (I) according to claim 1, wherein each of R₁, R₂ and R₃ is a hydrogen atom.

6. The compound of formula (I) according to claim 4, wherein each of R₄ and R₅ is a hydrogen atom.

7. The compound of formula ( I ) according to claim 4, wherein R₄ is a methyl group, and R₅ is a hydrogen atom.

8. The compound of formula (I) according to claim 4, wherein R₄ is a hydrogen atom, and R₅ is a methyl group.

9. The compound of formula (I) according to claim 4, wherein R₄ is a hydrogen atom, and R₅ is a halogen atom.

10. The compound of formula (I) according to claim 4, wherein R₄ is a hydrogen atom, and R₅ is a cyano group.

11. The compound of formula (I) according to claim 4, wherein R₄ is a hydrogen atom, and R₅ is a hydroxy group.

12. The compound of formula (I) according to claim 4, wherein R₄ is a hydrogen atom, and R₅ is selected from the group consisting of a carbamoyl group or a lower carbamoyl group.

13. The compound of formula (I) according to claim 5, wherein each of R₄ and R₅ is a hydrogen atom.

14. The compound of formula (I) according to claim 5, wherein R₄ is a methyl group, and R₅ is a hydrogen atom.

15. The compound of formula (I) according to claim 5, wherein R₄ is a hydrogen atom, and R₅ is a methyl group.

16. The compound of formula (I) according to claim 5, wherein R₄ is a hydrogen atom, and R₅ is a halogen atom.

17. The compound of formula (I) according to claim 5, wherein R₄ is a hydrogen atom, and R₅ is a cyano group.

18. The compound of formula (I) according to claim 5, wherein R₄ is a hydrogen atom, and R₅ is a hydroxy group.

19. The compound of formula (I) according to claim

5, wherein R₄ is a hydrogen atom, and R₅ is selected from the group consisting of a carbamoyl or a lower carbamoyl group.

20. The compound of formula (I) according to claim

6, wherein A is a halogen atom.

21. The compound of formula (I) according to claim 6, wherein A has the following formula (1) ;

R₆ is a hydrogen atom, R₇ is a lower alkanesulfonyl group, a N-(lower)alkylsulfamoyl, or a N,N-(lower)dialkylsulfamoyl group.9 22. The compound of formula (I) according to claim 6, wherein A has the following formula (1),

wherein R₆ is a methyl group, and R₇ is selected from the group consisting of a lower alkanesulfonyl group, a N-(lower)alkylsulfamoyl, or a N,N-(lower)dialkylsulfamoyl group.

23. The compound of formula (I) according to claim 6, wherein A has the following formula (2),

wherein R₆ is a hydrogen atom, and R₈ is a carbamoyl group which may be optionally substituted with appropriate substituent.

24. The compound of formula (I) according to claim 6, wherein A has the following formula (2),

wherein R₅ is a methyl group, and R₈ is a carbamoyl group which may be optionally substituted with appropriate substituent.

25. The compound of formula (I) according to claim

6, wherein A has the following formula (2) ;

wherein R₆ is a hydrogen atom, and _R8 is a heterocyclic group of 5- to 6- membered ring consisting of 1 to 4 heteroatoms selected from the group consisting of a nitrogen atom, an oxygen atom, or a sulfur atom which may be optionally substituted with substituents.

26. The compound of formula (I) according to claim 6, wherein A has the following formula (2) ;

wherein R₆ is a methyl group, and R₈ is a hetero cyclic group of 5- to 6- membered ring consisting of 1 to 4 heteroatoms selected from the group consisting of a nitrogen atom, an oxygen atom or a sulfur atom which may be optionally substituted with substituents.

27. The compound of formula (I) according to claim

7, wherein A has the following formula (1) ;

wherein R₅ is a hydrogen atom, and R₇ is selected from the group consisting of a lower alkanesulfonyl group, a N-(lower)alkylsulfamoyl, or a N,N-(lower)dialkylsulfamoyl group.

28. The compound of formula (I) according to claim 7, wherein A has the following formula (I),

29. The compound of formula (I) according to claim 7, wherein A has the following formula (2) ;

wherein R₆ is a hydrogen atom, and R₈ is a carbamoyl group which may be optionally substituted with appropriate substituents.

30. The compound of formula (I) according to claim 7, wherein A has the following formula (2) ;

wherein R₆ is a methyl group, and R₈ is a carbamoyl group which may be optionally substituted with appropriate substituents.

31. The compound of formula (I) according to claim 7, wherein A has the following formula (2) ;

wherein R₆ is a hydrogen atom, and R₈ is a heterocyclic group of 5- to 6- membered ring consisting of 1 to 4 heteroatoms selected from the group consisting of a nitrogen atom, an oxygen atom, or a sulfur atom, which may be optionally substituted with substituents.

32. The compound of formula (I) according to claim 7, wherein A has the following formula (2) ;

wherein R₆ is a methyl group, and R₈ is a heterocyclic group of 5- to 6- membered ring consisting of 1 to 4 heteroatoms selected from the group consisting of a nitrogen atom, an oxygen atom, or a sulfur atom, which may be optionally substituted with substituents.

33. The compound of formula (I) according to claim 8, wherein A has the following formula (1) ;

wherein R₆ is a hydrogen atom, and R₇ is selected from the group consisting of a lower alkanesulfonyl group, a N-(lower)alkylsulfamoyl, or a N,N-(lower)dialkylsulfamoyl group.

34. The compound of formula (I) according to claim 8, wherein A has the following formula (1) ;

wherein R₆ is a methyl group, and R₇ is selected from the group consisting of a lower alkanesulfonyl group, a N-(lower)alkyisulfamoyl, or a N,N-(lower)dialkylsulfamoyl group.

35. The compound of formula (I) according to claim 8, wherein A has the following formula (2) ;

36. The compound of formula (I) according to claim 8, wherein A has the following formula (2) ;

37. The compound of formula (I) according to claim 8, wherein A has the following formula (2) ;

38. The compound of formula (I) according to claim 8, wherein A has the following formula (2) ;

wherein R₆ is a hydrogen atom, and R₈ is a heterocyclic group of 5- to 6- membered ring consisting of 1 to 4 heteroatoms selected from the group consisting of a nitrogen atom, an oxygen atom, or a sulfur atom, which may be optionally substituted with substituents .

39. The compound of formula (I) according to claim 9, wherein A has the following formula (1) ;

40. The compound of formula (I) according to claim 9, wherein A has the following formula (1) ;

41. The compound of formula (I) according to claim 9, wherein A has the following formula (2) ;

wherein R₆ is a hydrogen atom, and R₈ is a carbamoyl group which may be optionally substituted with appropriate substitutents.

42. The compound of formula (I) according to claim 9, wherein A has the following formula (2) ;

wherein R₆ is a methyl group, and R₈ is a carbamoyl group which maybe optionally substituted with appropriate substitutents.

43. The compound of formula (I) according to claim 9, wherein A has the following formula (2) ;

wherein R₅ is a hydrogen atom, and R₈ is a heterocyclic group of 5- to 6- membered ring consisting of 1 to 4 heteroatoms selected from the group consisting of a nitrogen atom, an oxygen atom, or a sulfur atom, which may be optionally substituted with substitutents.

44. The compound of formula (I) according to claim 9, wherein A has the following formula (2) ;

45. The compound of formula (I) according to claim 10, wherein A has the following formula (1) ;

wherein R₆ is a hydrogen atom, and R₇ is selected from the group consisting of a lower alkanesulfonyl group, a N-(lower)alkylsulfamoyl, or a N,N- (lower)dialkylsulfamoyl group.

46. The compound of formula (I) according to claim 10, wherein A has the following formula (1) ;

wherein R₆ is a methyl group, and R₇ is selected from the group consisting of a lower alkanesulfonyl group, a N-(lower)alkylsulfamoyl, or a N,N- (lower)dialkylsulfamoyl group.

47. The compound of formula (I) according to claim 10, wherein A has the following formula (2) ;

48. The compound of formula (I) according to claim 10, wherein A has the following formula (2) ;

49. The compound of formula (I) according to claim 10, wherein A has the following formula (2) ;

50. The compound of formula (I) according to claim 10, wherein A has the following formula (2) ;

wherein R₆ is a methyl group, and R₈ is a heterocyclic group of 5- to 6- membered ring consisting of 1 to 4 heteroatoms selected from the group consisting of anitrogen atom, an oxygen atom, or a sulfur atom, which may be optionally substituted with substituents.

51. The compound of formula (I) according to claim 1, wherein the absolute configuration is selected from the group consisting of (5R, 6S, 8R, 2'S, 4'S) or (1R, 5S, 6S, 8R, 2'S, 4'S).

52. The compound according to claim 1, wherein the compound is (1R, 5S, 6S, 8R, 2'S, 4'S)-2-[[(E)-3-methanesulfonylamino-1-propenyl]pγrrolidin-4-ylthio]-6-(1-hydroxyethyl)-1-methyl-1-carbapen-2-em-3-carboxylic acid.

53. The compound according to claim 1, wherein the compound is (1R, 5S, 6S, 8R, 2'S, 4'S)-2-[[(E)-2-chloro-3-methanesulfonylamino-1-propenyl]pyrrolidin-4-yithio]- 6-(1-hydroxyethyl)-1-methyl-1-carbapen-2-em-3-carboxylic acid.

54. The compound according to claim 1, wherein the compound is (1R, 5S, 6S, 8R, 2'S, 4'S)-2-[[(E)-2-fluoro- 3-methanesulfonylamino-1-propenyl]pyrrolidine-4-yithio]- 6-(l-hydroxyethyl)-1-methyl-1-carbapen-2-em-3-carboxylic acid.

55. The compound according to claim 1, wherein the compound is (1R, 5S, 6S, 8R, 2'S, 4'S)-2-[[(E)-3-(N- methylsulfamoylamino-1-propenyl]pyrrolidin-4-ylthio]-6- (1-hydroxyethyl)-1-methyl-1-carbapen-2-em-3-carboxylic acid.

56. The compound according to claim 1, wherein the compound is (1R, 5S, 6S, 8R, 2'S, 4 'S)-2-[[(E)-3-(N,N-dimethylsulfamoylamino-1-propenyl]pyrrolidin-4-ylthio]-6-(1-hydroxyethyl)-1-methyl-1-carbapen-2-em-3-carboxylic acid.

57. The compound according to claim 1, wherein the compound is (1R, 5S, 6S, 8R, 2'S, 4'S)-2-[[(E)-2-chlorlo-3-(N-methylsulfamoylamino)-1-propenyl]pyrrolidin-4-ylthio] -6-(1-hydroxyethyl)-1-methyl-1-carbapen-2-em-3-carboxylic acid.

58. The compound according to claim 1, wherein the compound is (1R, 5S, 6S, 8R, 2'S, 4'S)-2-[[(E)-2-chlorlo-3-(N,N-dimethylsulfamoylamino)-l-propenyl]pyrrolidin-4-ylthio]-6-(1-hydroxyethyl)-1-methyl-1-carbapen-2-em-3-carboxylic acid.

59. The compound according to claim 1, wherein the compound is (1R, 5S, 6S, 8R, 2'S, 4'S)-2-[[(E)-2-fluoro-3-(N-methylsulfamoylamino)-1-propenyl]pyrrolidin-4-ylthio]-6-(1-hydroxyethyl)-1-methyl-1-carbapen-2-em-3-carboxylic acid.

60. The compound according to claim 1, wherein the compound is (1R, 5S, 6S, 8R, 2'S, 4'S)-2-[[(E)-2-fluoro- 3-(N,N-dimethylsufamoylamino)-1-propenyl]pyrrolidin-4-ylthio]-6-(1-hydroxyethyl)-1-methyl-1-carbapen-2-em-3-carboxylic acid.

61. The compound according to claim 1, wherein the compound is (1R, 5S, 6S, 8R, 2'S, 4'S)-2-[[(E)-3-methanesulfonylamino-2-methyl-1-propenyl]pyrrolidin-4- ylthio]-6-(1-hydroxyethyl)-1-methyl-1-carbapen-2-em-3- carboxylic acid.

62. The compound according to claim 1, wherein the compound is (1R, 5S, 6S, 8R, 2'S, 4'S)-2-[[(E)-2-methyl-3-(N-methylsulfamoyaImino)-1-propenyl]pyrrolidin-4-ylthio]-6-(1-hydroxyethyl)-1-methyl-1-carbapen-2-em-3-carboxylic acid.

63. The compound according to claim 1, wherein the compound is (1R, 5S, 6S, 8R, 2'S, 4'S)-2-[[(E)-3-(N,N-dimethylsulfamoylamino)-2-methyl-1-propenyl]pyrrolidin-4-ylthio]-6-(1-hydroxyethyl)-1-methyl-1-carbapen-2-em-3-carboxylic acid.

64. The compound according to claim 1, wherein the compound is (1R, 5S, 6S, 8R, 2'S, 4'S)-2-[[(E)-3-(4-morphorinosulfonylamino)-1-propenyl]pyrrolidin-4-ylthio]-6-(1-hydroxyethyl)-1-methyl-1-carbapen-2-ene-3-carboxylic acid.

65. The compound according to claim 1, wherein the compound is (1R, 5S, 6S, 8R, 2'S, 4'S)-2-[[(E)-2-methyl-3-(4-morphorinosulfonylamino)-1-propenyl]pyrrolidin-4-ylthio]-6-(1-hydroxyethyl)-1-methyl-1-carbapen-2-em-3-carboxylic acid.

66. The compound according to claim 1, wherein the compound is (1R, 5S, 6S, 8R, 2'S, 4'S)-2-[[(E)-3-(l-pyrrolidinosulfonylamino)-1-propenyl]pyrrolidin-4-ylthio]-6-(1-hydroxyethyl)-1-methyl-1-carbapen-2-em-3- carboxylic acid.

67. The compound according to claim 1, wherein the compound is (1R, 5S, 6S, 8R, 2'S, 4'S)-2-[[(E)-2-methyl- 3-(1-pyrrolidinosulfonylamino)-1-propenyl]prrolidin-4- ylthio]-6-(1-hydroxyethyl)-1-methyl-1-carbapen-2-em-3- carboxylic acid.

68. The compound according to claim 1, wherein the compound is (1R, 5S, 6S, 8R, 2'S, 4'S)-2-[[(E)-3-(1-piperazinosulfonylamino)-1-propenyl]pyrrolidin-4-ylthio]-6-(1-hydroxyethyl)-1-methyl-1-carbapen-2-em-3-carboxylic acid.

69. The compound according to claim 1, wherein the compound is (1R, 5S, 6S, 8R, 2'S, 4'S)-2-[[(E)-2-methyl-3-(1-piperazinosulfonylamino)-1-propenyl]pyrrolidin-4-ylthio] -6-(1-hydroxyethyl)-1-methyl-1-carbapen-2-em-3-carboxylic acid.

70. A process for preparing a compound of formula (IV)

or a pharmaceutically acceptable salt or ester thereof ; wherein R₁ is selected from the group consisting of a hydrogen atom or a methyl group, R₂ is selected from the group consisting of a metal or nonmetal salt group, or a carboxy protecting group, or a negative charge, R₃ is selected from the group consisting of hydrogen atom or an imino protecting group, or a pharmaceutically acceptable salt and R₄ and R₅ are each selected from the group consisting of a hydrogen atom, a hydroxy group, a cyano group, a halogen atom, methanesulfonyl group, N-methylsulfamoyl group, N,N-dimethylsulfamoyl group, or a lower alkyl group which may be optionally substituted with appropriate substituents, or a carbamoyl group which may be optionally substituted with appropriate substituents , and A is selected from the group co ns is ting o f a ha logen a t o m or the group o f th e following formula ( 1 ) or ( 2 ) ;

(wherein R₆ and R₇ are each selected from the group consisting of a hydrogen atom or a lower alkanesulfonyl group, a N-(lower)alkylsulfamoyl group, or a N,N-(lower) dialkylsulfamoyl group, wherein one of R₅ and R₇ is hydrogen, the other is other nonhydrogen group).

(wherein R₆ is the same as defined above, R₈ is selected from the group consisting of a carbamoyl group which may be optionally substituted with substituents, or a heterocyclic group of 5- to 6- membered ring consisting of 1 to 4 heteroatoms selected from the group consisting of a nitrogen atom, an oxygen atom, or a sulfur atom, which may be optionally substituted with substituents), R₉ is a hydrogen atom or a hydroxyi protecting group, and n is 0 to 3 ; which comprises reacting a compound of formula (II) ;

wherein R₁ is selected from the group consisting of a hydrogen atom or a methyl group, R₂ is selected from the group consisting of a metal or nonmetal salt group, or a carboxy protecting group, R₉ is selected from the group consisting of a hydrogen atom, a hydroxyl protecting group, or reactive derivatives thereof, with a compound of formula (III)

wherein R₃ is selected from the group consisting of a hydrogen atom or an iminoprotecting group, each of R₄ and R₅ is selected from the group consisting of a hydrogen atom, a hydroxy group, a cyano group, a halogen atom, a methanesulfonyl group, a N-methylsulfamoyl group, N,N-dimethylsulfamoyl group, or a lower alkyl group which may be optionally substituted with substituants, or a carbamoyl group which may be optionally substituted with appropriate substituents, and A is selected from the group consisting of a halogen atom or one of the following formula (1) - (2) ;

wherein R₆ and R₇ are each selected from the group consisting of a hydrogen atom, a lower alkanesulfonyl group, a N-(lower)alkylsulfamoyl group, or a N,N- (lower)dialkylsulfamoyl group, wherein either of R₆ or R₇ is hydrogen, the other group is nonhydrogen group.

wherein R₆ is the same as defined above, R₈ is selected from the group consisting of a carbamoyl group which may be optionally substituted with substituents or a heterocyclic group of 5- to 6- membered ring consisting of 1 to 4 heteroatoms selected from the group consisting of a nitrogen atom, an oxygen atom, or a sulfur atom, which may be optionally substituted with substituents and n is 0 to 3.

71. A process for preparing a compound of formula

(I)

wherein R₁ is selected from the group consisting of a hydrogen atom or a methyl group, R₂ is selected from the group consisting of a metal or nonmetal salt group, or a carboxy protecting group, R₃ is selected from the group consisting of a hydrogen atom or an lmmoprotec- ting group, each of R₄ and R₅ is selected from the group consisting of a hydrogen atom, a hydroxy group, a cyano group, or a halogen atom, a methanesulfonyl group, a N-methylsulfamoyl group, N,N-dimethylsulfamoyl group, or a lower alkyl group which may be optionally substituted with appropriate substituent, or a carbamoyl group which may be optionally substituted with appropriate substituent, and A is selected from the group consisting of a halogen atom, or one of the following formula (1) - (2) ;

wherein R₆ and R₇ are each selected from the group consisting of a hydrogen atom, a loweralkanesulfonyl group, a N-(lower)alkylsulfamoyl group, or a N,N-(lower)dialkylsulfamoyl group, and one of R₆ and R₇ is a hydrogen, the other group is nonhydrogen group.

wherein R₆ is the same as defined above, R₈ is selected from the group consisting of a carbamoyl group which may be optionally substituted with substitutent or a heterocyclic group of 5- to 6- membered ring consisting of 1 to 4 heteroatoms selected from the group consisting of a nitrogen atom, an oxygen atom, or a sulfur atom, which may be optionally substituted with substituents, and n is 0 to 3 or a pharmaceutically acceptable salt or ester thereof, and a pharmaceutically acceptable carrier or excipient ; by removing any protecting group of a compound of formula (IV);

72. A compound of claim 1 for use as a medicament.

73. A use of compounds of claim 2 for manufacturedicament for treatment of infectious disease.