WO1992002521A1

WO1992002521A1 - 1-azabicyclo[3.2.0]hept-2-ene-2-carboxylic acid compounds

Info

Publication number: WO1992002521A1
Application number: PCT/JP1991/000997
Authority: WO
Inventors: Masayoshi Murata; Toshiyuki Chiba; Hideo Tsutsumi; Akira Yamada; Kohji Hattori
Original assignee: Fujisawa Pharmaceutical Co., Ltd.
Priority date: 1990-07-27
Filing date: 1991-07-25
Publication date: 1992-02-20
Also published as: JPH06503803A

Abstract

A compound of formula (I) in which R1 is carboxy, protected carboxy or carboxylato, R2 is hydroxy(lower)alkyl or protected hydroxy-(lower)alkyl, R3 is axetidinyl, pyrrolidinyl, imidazolinyl, tetrahydropyrimidinyl, piperidyl, 1,1-di(lower)alkylpiperidinio, 3-(lower)-alkyl-1-(2-imidazolinio), 3-[hydroxy-(lower)alkyl]-1-(2-imidazolinio) or 3-(lower)alkyl-1,4,5,6-tetrahydro-1-pyrimidinio; wherein each heterocyclic group may be substituted by one or more suitable substituent(s), R10 is hydrogen or lower alkyl, and A is lower alkylene, provided that when R3 is lower alkylpyrrolidinyl, then R10 is hydrogen, or a pharmaceutically acceptable salt thereof, which is useful as an antimicrobial agent.

Description

DESCRIPTION

1-AZABICYCLO[3.2.0]HEPT-2-ENE-2-CARBOXYLIC ACID COMPOUNDS

The present invention relates to novel

1-azabicyclo[3.2.0]hept-2-ene-2-carboxylic acid compounds and pharmaceutically acceptable salts thereof.

More particularly, it relates to novel

1-azabicyclo[3.2.0]hept-2-ene-2-carboxylic acid compounds and pharmaceutically acceptable salts thereof, which have antimicrobial activity, to processes for the preparation thereof, to a pharmaceutical composition comprising the same, and to a use of the same as a medicament and in the treatment of infectious diseases caused by pathogenic microorganisms in human being or animal.

Accordingly, one object of the present invention is to provide novel 1-azabicyclo[3.2.0]hept-2-ene-2-carboxylic acid compounds and pharmaceutically acceptable salts thereof , which are highly active against a number of pathogenic microorganisms and are useful as antimicrobial agents. Another object of the present invention is to provide processes for the preparation of novel

1-azabicyclo[3.2.0]hept-2-ene-2-carboxylic acid compounds and salts thereof. A further object of the present invention is to provide a pharmaceutical composition comprising, as an active ingredient, said 1-azabicyclot 3.2.0]hept-2-ene-2-carboxylic acid compounds and pharmaceutically

acceptable salts thereof. Still further object of the present invention is to provide a use of said 1-azabicyclo[3.2.0]hept-2-ene- 2-carboxylic acid compounds and pharmaceutically

acceptable salts thereof as a medicament and in the treatment of infectious diseases caused by pathogenic microorganisms in human being or animal.

DISCLOSURE OF INVENTION The object 1-azabicyclo[3.2.0]hept-2-ene-2-carboxylic acid compounds are novel and can be represented by the following general formula :

in which R¹ is carboxy, protected carboxy or carboxylato,

R² is hydroxy(lower) alkyl or protected hydroxy¬

(lower)alkyl,

R³ is azetidinyl, pyrrolidinyl, imidazolinyl,

tetrahydropyrimidinyl, piperidyl,

1, 1-di(lower)alkylpiperidinio, 3-(lower)- alkyl-1-(2-imidazolinio), 3-[hydroxy¬

(lower)alkyl]-1-(2-imidazolinio) or

3-(lower)alkyl-1,4,5,6-tetrahydro-1- pyrimidinio;

wherein each heterocyclic group may be substituted by one or more suitable

substituent(s),

R¹⁰ is hydrogen or lower alkyl, and

A is lower alkylene,

provided that

when R³ is lower alkylpyrrolidinyl. then R¹⁰ is hydrogen,

or a pharmaceutically acceptable salt thereof.

In the object compound (I) and the intermediary compounds mentioned below, it is to be understood the there may be one or more stereo-isomeric pair(s) such as optical and/or geometrical isomers due to asymmetric carbon atom(s) and/or double bond(s), and such isomers are also included within the scope of the present invention.

Suitable salts of the object compound (I) are

conventional non-toxic, pharmaceutically acceptable salts and may include a salt with a base such as an inorganic base salt, for example, an alkali metal salt (e.g. sodium salt, potassium salt, etc.), an alkaline earth metal salt (e.g. calcium salt, magnesium salt, etc.), an ammonium salt, an organic base salt, for example, an organic amine salt (e.g. triethylamine salt, pyridine salt, picoline salt, ethanolamine salt, triethanolamine salt,

dicyclohexylamine salt, N,N'-dibenzylethylenediamine salt, etc.); a salt with an acid such as an inorganic acid addition salt (e.g. hydrochloride, hydrobromide, sulfate, phosphate, etc.), an organic acid addition salt (e.g.

formate, acetate, trifluoroacetate, maleate, tartrate, methanesulfonate, benzenesulfonate, etc.); a salt with a basic or acidic amino acid (e.g. arginine, aspartic acid, glutamic acid, etc.); and the like.

According to the present invention, the object compound (I) and pharmaceutically acceptable salts thereof can be prepared by the processes as illustrated by the following reaction schemes.

in which R¹, R², R³, R¹⁰ and A are each as defined above,

R_a ¹ is protected carboxy,

R_a ² is protected hydroxy(lower)alkyl, R_b ² is hydroxy(lower)alkyl,

R_a ³ is azetidmyl, pyrrolidinyl, imidazolinyl, tetrahydropyrimidinyl or piperidyl; wherein the ring carbon atom(s) may be substituted by one or more suitable substituent(s), and the ring nitrogen atom(s) is(are)

substituted by the imino-protective

group(s),

R_b ³ is azetidinyl, pyrrolidinyl, imidazolinyl, tetrahydropyrimidinyl or piperidyl; wherein the ring carbon atom(s) may be substituted by one or more suitable substituent(s), R_c ³ is azetidmyl, pyrrolidinyl, imidazolinyl, tetrahydropyrimidinyl or piperidyl; wherein the ring carbon atom(s) may be substituted by one or more suitable substituent(s), and the ring nitrogen atom(s) is(are)

substituted by lower alkanimidoyl or

N- (lower)alkyl(lower)alkanimidoyl, R_d ³ is azetidinyl, pyrrolidinyl, imidazolinyl, tetrahydropyrimidinyl or piperidyl; wherein the ring carbon atom(s) may be substituted by one or more suitable substituent(s), and the ring nitrogen atom(s) is (are) mono- or disubstituted by lower alkyl,

R⁴ is a suitable substituent(s),

R⁵ is lower alkyl or ar(lower)alkyl,

R⁶ is hydrogen or lower alkyl,

R⁹ is hydrogen, lower alkyl or

hydroxy(lower)alkyl,

R_a ⁹ is lower alkyl or hydroxy(lower)alkyl, and n is an integer of 2 or 3. Some of the starting compounds used in the Processes are new and can be prepared, for example, by the methods as shown in the following.

in which R¹, R², R³ , R⁹, R¹⁰, A and n are each as defined above,

R⁷ and R⁸ are each an amino-protective group, L is a leaving group, and

X is an acid residue.

In the above and subsequent descriptions of the present specification, suitable examples and illustrations of the various definitions which the present invention includes within the scope thereof are explained in detail as follows. The term "lower" is intended to mean 1 to 6 carbon atom(s), preferably 1 to 4 carbon atom(s), unless

otherwise indicated.

Suitable "protected carboxy" may include esterified carboxy, wherein "esterified carboxy" can be referred to the ones as mentioned below.

Suitable examples of the ester moiety of an

esterified carboxy may be the ones such as lower alkyl ester (e.g. methyl ester, ethyl ester, propyl ester, isopropyl ester, butyl ester, isobutyl ester t-butyl ester, pentyl ester, hexyl ester, etc.), which may have at least one suitable substituent(s), for example, lower alkanoyloxy(lower)alkyl ester [e.g. acetoxymethyl ester, propionyloxymethyl ester, butyryloxymethyl ester,

valeryloxymethyl ester, pivaloyloxymethyl ester,

hexanoyloxymethyl ester, 1-(or 2-)acetoxyethyl ester, 1-(or 2- or 3- ) acetoxypropy1 ester, 1-(or 2- or 3- or 4-)-acetoxybutyl ester, l-(or 2-)propionyloxyethyl ester, 1-(or 2- or 3-)propionyloxypropyl ester, 1-(or 2-)-butyryloxyethyl ester, 1-(or 2-)isobutyryloxyethyl ester, 1-(or 2-)pivaloyloxyethyl ester, 1-(or 2-)hexanoyloxyethyl ester, isobutyryloxymethyl ester, 2-ethylbutyryloxymethyl ester, 3,3-dimethylbutyryloxymethyl ester, 1-(or 2-)-pentanoyloxyethyl ester, etc.], lower alkanesulfonyl- (lower)alkyl ester (e.g. 2-mesylethyl ester, etc.), mono(or di or tri)halo(lower)alkyl ester (e.g. 2-iodoethyl ester, 2,2,2-trichloroethyl ester, etc.);

lower alkoxycarbonyloxy( lower)alkyl ester [e.g.

methoxycarbonyloxymethyl ester, ethoxycarbonyloxymethyl ester, propoxycarbonyloxymethyl ester,

t-butoxycarbonyloxymethyl ester, 1-(or 2-)methoxycarbonyl- oxyethyl ester, 1-(or 2-)ethoxycarbonyloxyethyl ester, 1-(or 2-)isopropoxycarbonyloxyethyl ester, etc.],

phthalidylidene(lower)alkyl ester, or (5-lower

alkyl-2-oxo-1,3-dioxol-4-yl) (lower)alkyl ester [e.g.

(5-methyl-2-oxo-1,3-dioxol-4-yl)methyl ester,

(5-ethyl-2-oxo-1,3-dioxol-4-yl)methyl ester,

(5-propyl-2-oxo-1,3-dioxol-4-yl)ethyl ester, etc.]; lower alkenyl ester (e.g. vinyl ester, allyl ester, etc.); lower alkynyl ester (e.g. ethynyl ester, propynyl ester, etc.); ar( lower)alkyl ester which may have at least one suitable substituent(s) (e.g. benzyl ester, 4-methoxybenzyl ester, 4-nitrobenzyl ester, phenethyl ester, trityl ester, benzhydryl ester, bis (methoxyphenyl)methyl ester,

3,4-dimethoxybenzyl ester, 4-hydroxy-3,5-di-t-butylbenzyl ester, etc.); aryl ester which may have at least one suitable substituent(s) (e.g. phenyl ester, 4-chlorophenyl ester, tolyl ester, t-butylphenyl ester, xylyl ester, mesityl ester, cumenyl ester, etc.); phthalidyl ester; and the like.

More preferable example of the protected carboxy thus defined may be C₂-C₄ alkenyloxycarbonyl and phenyl(or nitrophenyl) (C₁-C₄)alkoxycarbonyl, and the most preferable one may be allyloxycarbonyl.

Suitable "hydroxy(lower)alkyl" may include straight or branched lower alkyl substituted by hydroxy group such as hydroxymethyl, hydroxyethyl, hydroxypropyl, 1- (hydroxymethyl) ethyl, 1-hydroxy-1-methylethyl,

hydroxybutyl, hydroxypentyl, hydroxyhexyl, and the like, in which more preferable example may be

hydroxy(C₁-C₄) alkyl and the most preferable one may be hydroxymethyl, 2-hydroxyethyl and 1-hydroxyethyl.

Suitable "protected hydroxy(lower)alkyl" may include aforementioned hydroxy(lower)alkyl, in which the hydroxy group is protected by a conventional hydroxy-protective group such as those mentioned in the explanation of imino-protective group as mentioned below preferably, lower alkenyloxycarbonyl and phenyl(or nitrophenyl)- (lower)alkoxycarbonyl; and further C₆-C₁₀ ar(lower)alkyl such as mono- or di- or triphenyl(lower)alkyl (e.g.

benzyl, benzhydryl, trityl, etc.), etc.; trisubstituted silyl such as tri(lower)alkylsilyl (e.g. trimethylsilyl, triethylsilyl, isopropyldimethylsilyl,

t-butyldimethylsilyl, diisopropylmethylsilyl, etc.), tri(C₆-C₁₀)arylsilyl (e.g. triphenylsilyl, etc.),

tris[ (C₆-C₁₀) ar(lower)alkyl]silyl, for example,

tris[phenyl(lower)alkyl]silyl (e.g. tribenzylsilyl, etc.),etc.; and the like.

More preferable example of "protected

hydroxy(lower)alkyl" thus defined may be {phenyl(or nitrophenyl) (C₁-C₄)alkoxy}carbonyloxy(C₁-C₄) alkyl, C₂-C₄ alkenyloxycarbonyloxy(C₁-C₄) alkyl and

{tri(C₁-C₄)alkylsilyl}oxy(C₁-C₄)alkyl, and the most preferable one may be 1-t-butyldimethylsilyloxyethyl. Suitable "lower alkyl" may include straight or branched one such as methyl, ethyl, propyl, isopropyl, butyl, t-butyl, pentyl, hexyl, and the like, in which more preferable example may be C₁-C₄ alkyl and the most

preferable one may be methyl. Suitable "acid residue" may include an inorganic acid residue such as azide, halogen (e.g. chlorine, bromine, fluorine or iodine), and the like, an organic acid residue such as acyloxy (e.g. benzenesulfonyloxy, tosyloxy, methanesulfonyloxy, acetoxy, etc.), and the like, in which more preferable example may be halogen and the most preferable one may be chlorine.

Suitable "leaving group" may include acid residue as mentioned above, in which more preferable example may be lower alkanoyloxy and the most preferable one may be acetoxy.

Suitable "ar(lower)alkyl" may include C₆-C₁₀

ar( lower)alkyl such as phenyl(lower)alkyl (e.g. benzyl, phenethyl, etc.), tolyl(lower)alkyl, xylyl(lower)alkyl, naphthyl(lower)alkyl (e.g. naphthylmenthyl, etc.), and the like.

Suitable "imino-protective group" may include acyl such as aliphatic acyl, aromatic acyl, heterocyclic acyl and aliphatic acyl substituted by aromatic or heterocyclic group(s) derived from carboxylic, carbonic, sulfonic and carbamic acids.

The aliphatic acyl may include saturated or

unsaturated, acyclic or cyclic ones, for example, alkanoyl such as lower alkanoyl (e.g. formyl, acetyl, propionyl, butyryl, isobutyryl, valeryl, isovaleryl, pivaloyl, hexanoyl, etc.), alkylsulfonyl such as lower alkylsulfonyl (e.g. mesyl, ethylsulfonyl, propylsulfonyl,

isopropylsulfonyl, butylsulfonyl isobutylsulfonyl,

pentylsulfonyl, hexylsulfonyl, etc.), carbamoyl,

N-(lower)alkylcarbamoyl (e.g. methylcarbamoyl,

ethylcarbamoyl, etc.), alkoxycarbonyl such as lower alkoxycarbonyl (e.g. methoxycarbonyl, ethoxycarbonyl, propoxycarbonyl, butoxycarbonyl, t-butoxycarbonyl, etc.), alkenyloxycarbonyl such as lower alkenyloxycarbonyl (e.g. vinyloxycarbonyl, allyloxycarbonyl, etc.), alkenoyl such as lower alkenoyl (e.g. acryloyl, methacryloyl, crotonoyl, etc.), cycloalkanecarbonyl such as cyclo(lower)alkanecarbonyl (e.g. cyclopropanecarbonyl, cyclopentanecarbonyl, cyclohexanecarbonyl, etc.), and the like.

The aromatic acyl may include aroyl (e.g. benzoyl, toluoyl, xyloyl, etc.), N-arylcarbamoyl (e.g.

N-phenylcarbamoyl, N-tolylcarbamoyl, N-naphthylcarbamoyl, etc.), arenesulfonyl (e.g. benzenesulfonyl, tosyl, etc.), and the like.

The heterocyclic acyl may include heterocyclic-carbonyl (e.g. furoyl, thenoyl, nicotinoyl, isonicotmoyl, thiazolylcarbonyl, thiadiazolylcarbonyl, tetrazolylcarbonyl, etc.), and the like.

The aliphatic acyl substituted by aromatic group(s) may include aralkanoyl such as phenyl(lower)alkanoyl (e.g. phenylacetyl, phenylpropionyl, phenylhexanoyl, etc.), aralkoxycarbonyl such as phenyl(lower)alkoxycarbonyl (e.g. benzyloxycarbonyl, phenethyloxycarbonyl, etc.),

aryloxyalkanoyl such as phenoxytlower) alkanoyl (e.g.

phenoxyacetyl, phenoxypropionyl, etc.), and the like.

The aliphatic acyl substituted by heterocyclic group(s) may include heterocyclic-alkanoyl such as

heterocyclic(lower)alkanoyl (e.g. thienylacetyl,

imidazolylacetyl furylacetyl, tetrazolylacetyl,

thiazolylacetyl, thiadiazolylacetyl, thienylpropionyl, thiadaizolylpropionyl, etc.), and the like.

These acyl groups may be further substituted by one or more suitable substituents such as lower alkyl (e.g. methyl, ethyl, propyl, isopropyl, butyl, pentyl, hexyl, etc.), halogen (e.g. chlorine, bromine, iodine, fluorine), lower alkoxy (e.g. methoxy, ethoxy, propoxy, isopropoxy. butoxy, pentyloxy, hexyloxy, etc.), lower alkylthio (e.g. methylthio, ethylthio, propylthio, isopropylthio,

butylthio, pentylthio, hexylthio, etc.), nitro, lower alkylamino (e.g. methylamino, ethylamino, propylamino, isopropylamino, etc.), protected lower alkylamino (e.g.

N-allyloxycarbonyl-N-methylamino, etc.), and the like, and preferable acyl having such substituent(s) may be mono(or di or tri)haloalkanoyl (e.g. chloroacetyl, bromoacetyl, dichloroacetyl, trifluoroacetyl, etc.), mono(or di or tri)haloalkoxycarbonyl (e.g. chloromethoxycarbonyl, dichloromethoxycarbonyl, 2,2,2-trichloroethoxycarbonyl, etc.), nitro- (or halo- or lower alkoxy-)aralkoxycarbonyl (e.g. nitrobenzyloxycarbonyl, chlorobenzyloxycarbonyl, methoxybenzyloxycarbonyl, etc.), mono(or di or tri)-halo(lower)alkylsulfonyl (e.g. fluoromethylsulfonyl difluoromethylsulfonyl, trifluoromethylsulfonyl,

trichloromethylsulfonyl, etc.), and the like.

More preferable examples of imino-protective group thus defined may be C₁-C₄ alkanoyl,

(C₂-C₄) alkenyloxycarbonyl and phenyl(or

nitrophenyl) (C₁-C₄)alkoxycarbonyl, and the most preferable one may be acetyl and allyloxycarbonyl. Suitable "amino-protective group" may be the same as those for "imino-protective group", in which more

preferable examples of amino-protective group thus defined may be (C₂-C₄)alkenyloxycarbonyl and phenyl(or

nitrophenyl) (C₁-C₄)alkoxycarbonyl, and the most preferable one may be allyloxycarbonyl.

Suitable "azetidinyl, pyrrolidinyl, imidazolinyl, tetrahydropyrimidinyl, piperidyl, 1, 1-di(lower)-alkylpiperidinio, 3-(lower)alkyl-1-(2-imidazolinio),

3-[hydroxy(lower)alkyl]-1-(2-imidazolinio) or 3-(lower)alkyl-1,4,5,6-tetrahydro-1-pyrimidinio;

wherein each heterocyclic group may be substituted by suitable substituent(s)" may include : azetidmyl, pyrrolidinyl, imidazolinyl, tetrahydropyrimidinyl, piperidyl, 1,1-di(lower)alkylpiperidinio,

3-(lower)alkyl-1-(2-imidazolinio),

3-[hydroxy(lower)alkyl]-1-(2-imidazolinio) or 3- (lower) alkyl-1,4,5,6-tetrahydro-1-pyrimidinio;

wherein the ring carbon atom(s) may be substituted by one to three suitable substituent(s) selected from the group consisting of lower alkyl, pyridyl(lower)alkyl, carbamoyl, mono(or di) (lower)alkylcarbamoyl, carbamoyl(lower)alkyl, mono(or di) ( lower) alkylcarbamoyl(lower)alkyl,

hydroxy(lower)alkyl, lower alkoxy(lower)alkyl,

carbamoyloxy(lower)alkyl, mono(or

di) ( lower)alkylcarbamoyloxy(lower)alkyl, lower

alkylthio(lower)alkyl, lower alkylsulfinyl(lower)alkyl, lower alkenylthio(lower)alkyl, lower alkoxy(lower)-alkylthio(lower)alkyl, carbamoyloxy(lower)alkylthio(lower)alkyl, mono(or di) (lower) alkylcarbamoyl(lower)-alkylthio(lower)alkyl, lower alkylsulfonyl(lower)alkyl, mono(or di or tri)halo(lower) alkylthio(lower) alkyl, lower alkoxy(lower)alkanoylamino(lower)alkyl, lower

alkanoylamino(lower)alkyl, ureido(lower)alkyl, lower alkylsulfonylamino(lower)alkyl, lower alkoxycarbonylamino(lower)alkyl, (1-pyridinio) (lower)alkyl, 1- (lower)alkylpyridinio(lower)alkyl and N-containing heterocyclicthio(lower)alkyl wherein said heterocyclic group may be substituted by lower alkyl, and the ring nitrogen atom(s) may be substituted by the group consisting of lower alkyl, lower alkanimidoyl, N- ( lower) alkyl(lower)alkanimidoyl and imino-protective group.

Each explanation of the above-mentioned suitable substituent(s) can be referred to the ones as mentioned in the above and subsequent descriptions. Suitable "mono(or di) (lower)alkylcarbamoyl" means carbamoyl substituted by straight or branched lower alkyl as mentioned above, such as methylcarbamoyl,

dimethylcarbamoyl, ethylcarbamoyl, diethylcarbamoyl, N-methyl-N-ethylcarbamoyl, propylcarbamoyl,

dipropylcarbamoyl, isopropylcarbamoyl, butylcarbamoyl, pentylcarbamoyl, hexylcarbamoyl, and the like, in which more preferable example may be mono(or

di) (C₁-C₄) alkylcarbamoyl and the most preferable one may be dimethylcarbamoyl.

Suitable "carbamoyl(lower)alkyl" may include lower alkyl as mentioned above, wherein the lower alkyl is substituted with carbamoyl, in which more preferable example may be carbamoyl(C₁-C₄)alkyl and the most

preferable one may be carbamoylmethyl.

Suitable "mono ( or di ) ( lower ) alkylcarbamoyl( lower) -alkyl" means aforementioned carbamoyl(lower)alkyl, wherein the nitrogen atom(s) is(are) substituted by lower alkyl as mentioned above, in which more preferable example may be di(C₁-C₄)alkylcarbamoyl(C₁-C₄)alkyl and the most

preferable one may be N,N-dimethylcarbamoylmethyl. Suitable "pyridyl(lower)alkyl" may include lower alkyl as mentioned above, wherein the lower alkyl is substituted with pyridine, in which more preferable example may be pyridyl(C₁-C₄)alkyl and the most preferable one may be pyridin-3-ylmethyl.

Suitable "lower alkoxy(lower)alkyl" means

aforementioned lower alkyl substituted by lower alkoxy such as methoxy, ethoxy, propoxy, isopropoxy, butoxy, pentyloxy, hexyloxy, and the like, in which the most preferable example may be methoxymethyl and methoxyethyl. Suitable carbamoyloxy(lower)alkyl" means

aforementioned lower alkyl substituted by carbamoyloxy, in which the most preferable example may be

carbamoyloxymethyl and 2-carbamoyloxyethyl.

Suitable "mono(or di) (lower) alkylcarbamoyloxy(lower)- alkyl" means aforementioned lower alkyl substituted by mono(or di) (lower)alkylcarbamoyloxy, wherein the mono(or di) ( lower) alkylcarbamoyl moiety may be the same as those mentioned above, in which the most preferable example may be N-ethylcarbamoyloxymethyl.

Suitable "lower alkylthio(lower)alkyl" means

aforementioned lower alkyl substituted by lower alkylthio such as methylthio, ethylthio, propylthio, isopropylthio, butylthio, pentylthio, hexylthio, and the like, in which the most preferable example may be methylthiomethyl and t-butylthiomethyl. Suitable "lower alkylsulfinyl(lower)alkyl" means aforementioned. lower alkyl substituted by lower

alkylsulfinyl such as methylsulfinyl, ethylsulfinyl, propylsulfinyl, isopropylsulfinyl, butylsulfinyl,

pentylsulfinyl, hexylsulfinyl, and the like, in which the most preferable example may be methylsulfinylmethyl.

Suitable "lower alkenylthio(lower) alkyl" means aforementioned lower alkyl substituted by lower

alkenylthio such as allylthio, vinylthio, and the like, in which the most preferable example may be allylthiomethyl.

Suitable "lower alkoxy( lower)alkylthio(lower)alkyl" means aforementioned lower alkyl substituted by lower alkoxy(lower)alkylthio wherein the lower

alkoxy(lower)alkyl moiety may be the same as mentioned above, in which the most preferable example may be methoxymethylthiomethyl and 2-methoxyethylthiomethyl.

Suitable "carbamoyloxy(lower)alkylthio(lower)alkyl" means aforementioned lower alkyl substituted by

carbamoyloxy(lower)alkylthio wherein the

carbamoyloxy(lower) alkyl moiety may be the same as those mentioned above, in which the most preferable example may be carbamoyloxymethylthiomethyl.

Suitable "mono(or di) (lower)alkylcarbamoyl(lower)-alkylthio(lower)alkyl" means aforementioned lower alkyl substituted by mono(or di) (lower)alkylcarbamoyl(lower)- alkylthio wherein the mono(or di) (lower)alkylcarbamoyl-(lower) alkyl moiety may be the same as those mentioned above, in which the most preferable example may be

N,N-dimethylcarbamoylmethylthiomethyl.

Suitable "lower alkanoylamino(lower)alkyl" means aforementioned lower alkyl substituted by lower

alkanoylamino wherein the lower alkanoylamino moiety may be the same as mentioned in the explanation of the acyl group, in which the most preferable example may be

acetylaminomethyl.

Suitable "ureido(lower) alkyl" means aforementioned lower alkyl substituted by ureido, in which the most preferable example may be ureidomethyl.

Suitable "lower alkylsulfonylamino(lower)alkyl" means aforementioned lower alkyl substituted by lower

alkylsulfonylamino such as mesylamino, ethylsulfonylamino, propylsulfonylamino, isopropylsulfonylamino,

butylsulfonylamino, pentylsulfonylamino,

hexylsulfonylamino, and the like, in which the most preferable example may be mesylsulfonylaminomethyl.

Suitable "lower alkoxycarbonylamino(lower)alkyl" means aforementioned lower alkyl substituted by lower alkoxycarbonylamino wherein the lower alkoxycarbonyl moiety may be the same as those mentioned in the

explanation of protected carboxy, in which the most preferable example may be methoxycarbonylaminomethyl. Suitable "(1-pyridinio) (lower)alkyl" means

aforementioned lower alkyl substituted by 1-pyridinio, in which the most preferable example may be

3-(1-pyridinio)propyl. Suitable "1-(lower)alkylpyridinio(lower)alkyl"means aforementioned lower alkyl substituted by

1-(lower)alkylpyridinio, in which the most preferable example may be 3- (1-methyl-3-pyridinio)propyl. Suitable "lower alkanimidoyl" may include straight or branched one such as formimidoyl, acetimidoyl,

propionimidoyl, butyrimidoyl, valerimidoyl,

isovalerimidoyl, hexanimidoyl, and the like, in which the most preferable example may be formimidoyl and

acetimidoyl.

Suitable "N-(lower)alkyl(lower)alkanimidoyl" means aforementioned lower alkanimidoyl substituted by lower alkyl at the nitrogen atom, in which the most preferable example may be N-methylformimidoyl.

Suitable "N-containing heterocyclic group" in the term of "optionally substituted N-containing

heterocyclicthio(lower)alkyl" means saturated or

unsaturated, monocyclic or polycyclic heterocyclic group containing at least one nitrogen atom and optionally other hetero-atom(s) such as an oxygen, sulfur, nitrogen atom and the like.

Preferable N-containing heterocyclic group moiety may be heterocyclic group such as :

- unsaturated 3 to 8-membered, preferably 5 or

6-membered heteromonocyclic group containing 1 to 4 nitrogen atom(s), for example, pyrrolyl, pyrrolinyl, imidazolyl, pyrazolyl, pyridyl, and its N-oxide,

pyrimidyl, pyrazinyl, pyridazinyl, triazolyl (e.g.,

4H-1,2,4-triazolyl, 1H-1,2,3-triazolyl,

2H-1,2,3-triazolyl, etc.), tetrazolyl (e.g.,

1H-tetrazolyl, 2H-tetrazolyl, etc.), dihydrotriazinyl (e.g., 4,5-dihydro-1,2,4-triazinyl,

2,5-dihydro-1,2,4-triazinyl, etc.), etc.;

- saturated 3 to 8-membered, preferably 5 or

6-membered heteromonocyclic group containing 1 to 4 nitrogen atom(s), for example, azetidinyl, pyrrolidinyl, imidazolidinyl, piperidinyl, pyrazolidinyl, piperazinyl, etc.;

- unsaturated condensed 7 to 12-membered heterocyclic group containing 1 to 5 nitrogen atom(s), for example, indolyl, isoindolyl, indolizinyl, benzimidazolyl,

guinolyl, isoquinolyl, indazolyl, benzotriazolyl,

tetrazolopyridyl, tetrazolopyridazinyl (e.g.,

tetrazolo[1,5-b]pyridazinyl, etc.),

dihydrotriazolopyridazinyl, etc.;

- unsaturated 3 to 8-membered, preferably 5 or

6-membered heteromonocyclic group containing 1 to 2 oxygen atom(s) and 1 to 3 nitrogen atom(s), for example,

oxazolyl, isoxazolyl, oxadiazolyl, (e.g.,

1,2,4-oxadiazolyl, 1,3,4-oxadiazolyl, 1,2,5-oxadiazolyl, etc.), etc.;

- saturated 3 to 8-membered, preferably 5 or

6-membered heteromonocyclic group containing 1 to 2 oxygen atom(s) and 1 to 3 nitrogen atom(s), for example, morpholinyl, etc.;

- unsaturated condensed 7 to 12-membered heterocyclic group containing 1 to 2 oxygen atom(s) and 1 to 3 nitrogen atom(s), for example, benzoxazolyl, benzoxadiazolyl, etc.;

- unsaturated 3 to 8-membered, preferably 5 or

6-membered heteromonocyclic group containing 1 to 2 sulfur atom(s) and 1 to 3 nitrogen atom(s), for example,

1,3-thiazolyl, 1,2-thiazolyl, thiazolinyl, thiadiazolyl (e.g., 1,2,4-thiadiazolyl, 1,3,4-thiadiazolyl,

1,2,5-thiadiazolyl, 1,2,3-thiadiazolyl), etc.;

- saturated 3 to 8-membered, preferably 5 or

thiazolidinyl, etc.;

- unsaturated condensed 7 to 12-membered heterocyclic group containing 1 to 2 sulfur atom(s) and 1 to 3 nitrogen atom(s), for example, benzothiazolyl, benzothiadiazolyl, etc. and the like;

wherein said heterocyclic group is optionally

substituted by one or more, preferably one or two suitable substituent(s) such as :

- hydroxy,

- protected hydroxy, in which the hydroxy group is protected by a conventional hydroxy-protective group as mentioned in the explanation of protected

hydroxy(lower)alkyl,

- hydroxy(lower)alkyl or protected

hydroxy(lower)alkyl as mentioned above,

- halogen (e.g. chlorine, bromine, iodine or

fluorine),

- lower alkoxy as mentioned above,

- lower alkyl as mentioned above (e.g. methyl, etc.),

- lower alkoxy(lower)alkyl as mentioned above,

- lower alkylamino, in which the lower alkyl moiety may be the same as those for lower alkyl as mentioned above,

- protected lower alkylamino, which is the lower alkylamino group as mentioned above, in which the amino group is protected by a conventional amino-protective group as mentioned above.

Preferable example of "optionally substituted

N-containing heterocyclicthio(lower)alkyl" thus defined may be lower alkyl substituted by unsaturated 5 or 6 membered heteromonocyclic group containing 1 to 4 nitrogen atom(s) such as pyrrolyl, pyrrolinyl, imidazolyl,

pyrazolyl, pyridyl, and its N-oxide, pyrimidyl, pyrazinyl, pyridazinyl, triazolyl, tetrazolyl, dihydrotriazinyl, wherein said heterocyclic group is optionally substituted by lower alkyl, in which more preferable example may be lower alkylimidazolyl(lower)alkyl and lower

alkyltetrazolyl(lower)alkyl, and the most preferable example may be 1-methylimidazol-2-yl and

1-methyltetrazol-5-yl.

Suitable "lower alkylsulfonyl(lower)alkyl" means aforementioned lower alkyl substituted by lower

alkylsulfonyl such as methylsulfonyl, ethylsulfonyl, propylsulfonyl, butylsulfonyl, pentylsulfonyl,

hexylsulfonyl, and the like, in which the most preferable example may be methylsulfonylmethyl.

Suitable "mono(or di or

tri)halo(lower)alkylthio(lower)alkyl" means aforementioned lower alkyl substituted by halo(lower)alkylthio, wherein the halo(lower)alkyl moiety may be straight or branched lower alkyl having one to three halogen (e.g. chlorine, bromine, iodine, fluorine) such as chloromethyl, fluoromethyl, dichloromethyl, dibromomethyl, diiodomethyl, difluoromethyl, trifluoromethyl, chloroethyl, chlorofluoroethyl, difluoroethyl, trifluoroethyl, chloropropyl, difluoropropyl, trichlorobutyl,

chloropentyl, chlorohexyl, and the like, in which the most preferable example may be difluoromethylthiomethyl.

Suitable "lower alkoxy(lower)alkanoylamino- (lower)alkyl" means aforementioned lower alkyl substituted by lower alkoxy(lower)alkanoylamino wherein the lower alkoxy and lower alkanoyl moieties may be the same as those mentioned above, in which the most preferable example may be methoxyacetylaminomethyl.

Suitable "3-(lower)alkyl-1-(2-imidazolinio)" means 2-imidazolin-1-yl substituted by aforementioned lower alkyl at 3-position, in which the most preferable example may be 3-methyl-1-(2-imidazolinio).

Suitable "3-[hydroxy(lower)alkyl]-1-(2-imidazolinio)" means 2-imidazolin-1-yl substituted by aforementioned hydroxy(lower) alkyl at 3-position, in which the most preferable example may be 3-(2-hydroxyethyl)-1-(2-imidazolinio).

Suitable "3-(lower)alkyl-1,4,5,6-tetrahydro-1-pyrimidinio" means 1,4,5,6-tetrahydropyrimidin-1-yl substituted by .aforementioned lower alkyl at 3-position, in which the most preferable example may be 3-methyl-1,4,5,6-tetrahydro-1-pyrimidinio.

Suitable "1,1-di(lower)alkylpiperidinio"means

piperidinyl disubstituted by aforementioned lower alkyl at 1-position, in which the most preferable example may be 1, 1-dimethyl-4-piperidinio. Preferable example of R³ may be :

- imidazolinyl [e.g. 2-imidazolin-1(or 4)-yl, etc.];

- lower alkylimidazolinyl such as 2-(C₁-C₄)alkyl-2-imidazolinyl [e.g. 2-methyl-2-imidazolin-1(or 4)-yl, etc.], etc.;

- [carbamoyl(lower)alkyl]imidazolinyl such as

2-carbamoyl(C₁-C₄)alkyl-2-imidazolinyl [e.g. 2-carbamoylmethyl-2-imidazolin-1(or 4)-yl, etc.], etc.;

- [N,N-di(lower)alkylcarbamoyl(lower)alkyl]¬imidazolinyl such as 2-[N,N-di(C₁-C₄)alkylcarbamoyl(C₁-C₄) alkyl]-2-imidazolinyl [e.g. 2-(N,N- dimethylcarbamoylmethyl)-2-imidazolin-4-yl, etc.], etc.;

- [pyridyl(lower)alkyl]imidazolinyl such as 2-{3-pyridyl(C₁-C₄)alkyl}-2-imidazolinyl [e.g.

2-(3-pyridylmethyl)-2-imidazolin-1(or 4)-yl, etc.], etc.;

- azetidinyl (e.g. azetidin-3-yl, etc.);

- 1-(lower)alkenyloxycarbonylazetidinyl such as 1- (C₂-C₄)alkenyloxycarbonylazetidinyl (e.g.

1-allyloxycarbonylazetidin-3-yl, etc.);

- 1-(lower)alkanimidoylazetidinyl such as

1- ( C₁-C₄)alkanimidoylazetidinyl (e.g.

1-formimidoylazetidin-3-yl, etc.);

- pyrrolidinyl (e.g. pyrrolidin-2(or 3 )-y1, etc.);

- 1- ( lower)alkenyloxycarbonylpyrrolidinyl such as 1-(C₂-C₄)alkenyloxycarbonylpyrrolidinyl (e.g.

1-allyloxycarbonylpyrrolidin-2(or 3 )-yl, etc.);

- 1-(lower)alkanimidoylpyrrolidinyl such as

1-(C₁-C₄) alkanimidoylpyrrolidinyl (e.g.

1-formimidoylpyrrolidin-2-yl, etc.);

- 1-[N-(lower)alkyl(lower)alkanimidoyl]pyrrolidinyl such as 1-[N-(C₁-C₄)alkyl(C₁-C₄)alkanimidoyl]pyrrolidinyl [e.g. 1-(N-methylformimidoyl)pyrrolidin-2-yl, etc.];

- [N,N-di(lower)alkylcarbamoyl]pyrrolidinyl such as [N,N-di(C₁C₄)alkylcarbamoyl]pyrrolidinyl [e.g. 2-(N,N-dimethylcarbamoyl)pyrrolidin-4-yl, etc.];

- 1-(lower)alkanimidoyl-[N,N-di(lower)alkylcarbamoyl]pyrrolidinyl such as 1-(C₁-C₄)alkanimidoyl-2- [N,N-di(C₁-C₄)alkylcarbamoyl]pyrrolidinyl [e.g.

1-formimidoyl-2-(N,N-dimethylcarbamoyl)pyrrolidin-4-yl, etc.], etc.;

- [hydroxy(lower)alkyl]imidazolinyl such as

[hydroxy(C₁-C₄)alkyl]-2-imidazolinyl [e.g. 2-(hydroxymethyl)-2-imidazolin-4-yl,

2-(2-hydroxyethyl)-2-imidazolin-4-yl, etc.];

- [lower alkoxy(lower)alkyl]imidazolinyl such as C₁-C₄ alkoxy(C₁-C₄)alkyl]-2-imidazolinyl [e.g.

2-(methoxymethyl)-2-imidazolin-4-yl, 2-(2-methoxyethyl)-2- imidazolin-4-yl, etc.];

- [carbamoyloxy(lower) alkyl]imidazolinyl such as

[carbamoyloxy(C₁-C₄)alkyl]-2-imiazolinyl [e.g.

2-(carbamoyloxyethyl)-2-imidazolin-4-yl,

2-(carbamoyloxymethyl)-2-imidazolin-4-yl, etc.];

- [mono or di(lower) alkylcarbamoyloxy(lower)alkyl]- imidazolinyl such as N-(C₁-C₄) alkylcarbamoyloxy(C₁-C₄)- alkyl-2-imidazolinyl [e.g. 2-(N-ethylcarbamoyloxymethyl)- 2-imidazolin-4-yl, etc.];

- [lower alkylthio(lower)alkyl]imidazolinyl such as

[C₁-C₄ alkylthio(C₁-C₄) alkyl]-2-imidazolinyl [e.g.

2-(methylthiomethy1)-2-imidazolin-4-y1,

2- (t-butylthiomethyl)-2-imidazolin-4-yl, etc.];

- [lower alkylsulfinyl(lower)alkyl]imidazolinyl such as [C₁-C₄ alkylsulfinyl(C₁-C₄)alkyl]-2-imidazolinyl [e.g.

2-(methylsulfinylmethyl)-2-imidazolin-4-yl, etc.];

- [lower alkenylthio(lower)alkyl]imidazolinyl such as [C₂-C₄ alkenylthio(C₁-C₄)alkyl]-2-imidazolinyl [e.g. 2-

(allylthiomethyl)-2-imidazolin-4-yl, etc.];

- [lower alkoxy(lower)alkylthio(lower)alkyl]-imidazoliinyl such as [C₁-C₄ alkoxy(C₁-C₄)alkylthio(C₁-C₄)-alkyl]-2-imidazolinyl [e.g. 2-(2-methoxyethylthiomethyl)-2-imidazolin-4-yl, etc.];

- [carbamoyloxy(lower)alkylthio(lower)alkyl]- imidazolinyl such as [carbamoyloxy(C₁-C₄)alkylthio(C₁-C₄)-alkyl]-2-imidazolinyl [e.g. 2-(2-carbamoyloxyethyl)thiomethyl-2-imidazolin-4-yl, etc.];

- [mono or di(lower)alkylcarbamoyl(lower)alkylthio- ( lower)alkyl]imidazolinyl such as [N,N-di(C₁-C₄)- alkylcarbamoyl(C₁-C₄)alkylthio(C₁-C₄)alkyl]-2-imidazolinyl [e.g. N,N-dimethylcarbamoylmethylthiomethyl -2-imidazolin-4-yl, etc. ] ;

- [lower alkylimidazolylthio(lower)alkyl]imidazolinyl such as [C₁-C₄ alkylimidazolylthio(C₁-C₄)alkyl]-2-imidazolinyl [e.g. 2-(1-methylimidazol-2-ylthiomethyl)-2- imidazolin-4-yl, etc.];

- [ lower alkyltetrazolylthio(lower)alkyl]imidazolinyl such as [C₁-C₄ alkyltetrazolylthio(C₁-C₄)alkyl]-2- imidazolinyl [e.g. 2-(1-methyltetrazol-5-ylthiomethyl)-2-imidazolin-4-yl, etc.];

- [lower alkanoylamino(lower)alkyl]imidazolinyl such as [C₁-C₄ alkanoylamino(C₁-C₄)alkyl]-2-imidazolinyl [e.g. 2-(acetylaminomethyl)-2-imidazolin-4-yl, etc.];

- [ureido(lower)alkyl]imidazolinyl such as

[ureido(C₁-C₄)alkyl]-2-imidazolinyl [e.g.

2-(ureidomethyl)-2-imidazolin-4-yl, etc.];

- [ lower alkylsulfonylamino(lower)alkyl]imidazolinyl such as [C₁-C₄ alkylsulfonylamino(C₁-C₄)alkyl]-2-imidazolinyl [e.g. 2-(mesylaminomethyl)-2-imidazolin-4-yl, etc.];

- [lower alkoxycarbonylamino(lower)alkyl]imidazolinyl such as [C₁-C₄.alkoxycarbonylamino(C₁-C₄)alkyl]-2- imidazolinyl [e.g. 2-(methoxycarbonylaminomethyl)-2-imidazolin- 4-yl, etc.];

- [lower alkyl] [lower alkoxy(lower)alkyl]imidazolinyl such as N-[C₁-C₄ alkyl]-[C₁-C₄ alkoxy(C₁-C₄)alkyl]-2- imidazolinyl [e.g. 1-methyl-2-(methoxymethyl)-2- imidazolin-5-yl, etc. ];

- N-[lower alkanoyl] [carbamoyl(lower)alkyl]- imidazolinyl such as N-[C₁-C₄ alkanoyl]-[carbamoyl(C₁-C₄)-alkyl]-2-imidazolinyl [e.g. 1-acetyl-2-(carbamoylmethyl)- 2-imidazolin-4-(or 5)-yl, etc.];

- tetrahydropyrimidinyl such as 1,4,5,6-tetrahydropyrimidinyl (e.g. 1,4,5,6-tetrahydropyrimidin-1-yl, etc.);

- (lower alkyl)tetrahydropyrimidinyl such as (C₁-C₄ alkyl)-1,4,5,6-tetrahydropyrimidinyl [e.g. 2-methyl- 1,4,5,6-tetrahydropyrimidin-1-yl, etc.];

- [pyridinio(lower) alkyl]imidazolinyl such as

[ (1-pyridinio) (C₁-C₄)alkyl]-2-imidazolinyl [e.g.

2-{3-(1-pyridinio)propyl}-2-imidazolin-4-yl, etc.];

- [1-(lower)alkylpyridinio(lower)alkyl]imidazolinyl such as [1-(C₁-C₄)alkylpyridinio(C₁-C₄)alkyl]-2-imidazolinyl [e.g. 2-(1-methyl-3-pyridinio)methyl-2-imidazolin-1(or 4)-yl, etc.];

- 3-[hydroxy(lower)alkyl]-1-(2-imidazolinio) such as 3-[hydroxy(C₁-C₄)alkyl]-1-(2-imidazolinio) [e.g.

3- ( 2-hydroxyethyl)-1-(2-imidazolinio), etc.];

- 2 ,3-di(lower)alkyl-1-(2-imidazolinio) such as 2 , 3-di(C₁-C₄)alkyl-1-(2-imidazolinio) [e.g.

2 , 3-dimethyl-1-(2-imidazolinio), etc.];

- 3-(lower)alkyl-1,4,5,6-tetrahydro-1-pyrimidinio optionally substituted by lower alkyl such as

3-(C₁-C₄)alkyl-1,4,5,6-tetrahydro-1-pyrimidinio optionally substituted by C₁-C₄ alkyl [e.g.

3-methyl-1,4,5,6-tetrahydro-1-pyrimidinio, 2,3-dimethyl- 1,4,5,6-tetrahydro-1-pyrimidinio, etc.];

- 1-1-di(lower)alkylpiperidinio such as

1, 1-di( C₁-C₄) alkylpiperidinio (e.g.

1, 1-dimethyl-4-piperidinio, etc.);

- [lower alkylsulfonyl(lower)alkyl]imidazolinyl such as [C₁-C₄ alkylsulfonyl(C₁-C₄)alkyl]-2-imidazolinyl [e.g.

2- (methylsulfonylmethyl)-2-imidazolin-4-yl, etc.];

- [dihalo( lower)alkylthio(lower)alkyl]imidazolinyl such as [dihalo(C₁-C₄) alkylthio(C₁-C₄)alkyl]-2-imidazolinyl [e.g. 2-(difluoromethylthiomethyl)-2-imidazolin-4-yl, etc.];

- [ lower alkoxy(lower)alkanoylamino(lower)alkyl]imidazolinyl such as [C₁-C₄ alkoxy(C₁-C₄)alkanoylamino(C₁-C₄) alkyl]-2-imidazolinyl [e.g. 2-(methoxyacetylaminomethyl)-2-imidazolin-4-yl, etc.];

- piperidyl [e.g. 4-piperidyl, etc.];

- [lower alkoxy(lower)alkyl]pyrrolidinyl such as [C₁-C₄ alkoxy(C₁-C₄)alkyl]pyrrolidinyl [e.g.

2-methoxymethylpyrrolidin-3-( or 4)-yl, etc.];

- [hydroxy(lower)alkyl]pyrrolidinyl such as

[hydroxy(C₁-C₄)alkyl]pyrrolidinyl [e.g. 2-(hydroxymethyl)- pyrrolidin-4-yl, etc.];

- 1-( lower alkanimidoyl)piperidyl such as

1- (C₁-C₄)alkanimidoypiperidyl (e.g.

1-formimidoyl-4-piperidyl, etc.).

Suitable "lower alkylene" may include straight or branched one such as methylene, ethylene, trimethylene, tetramethylene, pentamethylene, hexamethylene,

methylmethylene, ethylethylene, propylene, and the like, in which more preferable example may be C₁-C₄ alkylene and the most preferable one may be methylene and ethylene. Among the members of R³, as to the imidazolinyl ring of this invention, it is easily understood that there are tautomeric isomers as shown by the following equilibriums:

All of the above tautomeric isomers are included within the scope of the present invention, and in the present invention, however, said imidazolinyl ring in the object and intermediate compounds is represented by using one of the expressions therefor and a nomenclature

corresponding to it.

The processes for the preparation of the object compound (I) of the present invention are explained in detail in the following. ( 1 ) Process 1 :

The compound (I) or salts thereof can be prepared by cyclizing the compound (II) or its reactive derivative at the carbonyl group adjacent to R¹ or salts thereof.

Suitable salts of the compound (II) may be the same as those for the compound (I).

Suitable reactive derivative of the compound (II) may include tri(lower)alkoxyphosphoranylidene compound thereof (e.g. triethoxyphosphoranylidene compound, etc.),

triarylphosphoranylidene compound thereof (e.g.

triphenylphosphoranylidene compound, etc.), etc., each of which can be prepared by reacting the compound (II) with tri(lower)alkyl phosphite or triarylphosphine,

respectively.

This reaction is preferably carried out by heating the compound (II) in a conventional solvent which does not adversely influence the reaction such as dioxane,

hexamethylphosphoramide, benzene, toluene, xylene,

dimethyl sulfoxide, N,N-dimethylformamide, pyridine, etc., or a mixture thereof. This reaction can also be carried out in the presence of hydroquinone.

The reaction temperature of this reaction is not critical and the reaction is usually carried out under from warming to heating.

(2) Process 2 :

The compound (I-b) or salts thereof can be prepared by subjecting the compound (I-a) or salts thereof to removal reaction of the carboxy-protective group. Suitable salts of the compounds (I-a) and (I-b) may be the same as those for the compound (I) .

The present reaction is usually carried out by a conventional method such as hydrolysis, reduction, and the like.

(i) Hydrolysis :

Hydrolysis is preferably carried out in the presence of a base or an acid. Suitable base may include an alkalimetal hydroxide (e.g. sodium hydroxide, potassium hydroxide, etc.), an alkaline earth metal hydroxide (e.g. magnesium hydroxide, calcium hydroxide, etc.), alkali metal hydride (e.g. sodium hydride, potassium hydride, etc.), alkaline earth metal hydride (e.g. calcium hydride, etc.), alkali metal alkoxide (e.g. sodium methoxide, sodium ethoxide, potassium t-butoxide, etc.), an alkali metal carbonate (e.g. sodium carbonate, potassium

carbonate, etc.), and alkaline earth metal carbonate (e.g. magnesium carbonate, calcium carbonate, etc.), an alkali metal bicarbonate (e.g. sodium bicarbonate, potassium bicarbonate, etc.), and the like.

Suitable acid may include an organic acid (e.g.

formic acid, acetic acid, propionic acid, trifluoroacetic acid, benzenesulfonic acid, p-toluenesulfonic acid, etc.) and an inorganic acid (e.g. hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, etc.). The acidic hydrolysis using trifluoroacetic acid is usually

accelerated by addition of cation trapping agent (e.g.

phenol, anisole, etc.).

In case that the hydroxy-protective group is

tri ( lower)alkylsilyl, the hydrolysis can be carried out in the presence of tri( lower)alkylammonium halide (e.g.

tributylammonium fluoride, etc.). This reaction is usually carried out in a

conventional solvent which does not adversely influence the reaction such as water, dichloromethane, alcohol (e.g. methanol, ethanol, etc.), tetrahydrofuran, dioxane, acetone, etc., or a mixture thereof. A liquid base or acid can be also used as the solvent.

The reaction temperature is not critical and the reaction is usually carried out under from cooling to heating.

( ii) Reduction :

The reduction method applicable for this removal reaction may include, for example, reduction by using a combination of a metal (e.g. zinc, zing amalgam, etc.) or a salt of chrome compound (e.g. chromous chloride,

chromous acetate, etc.) and an organic or inorganic acid (e.g. acetic acid, propionic acid, hydrochloric acid, sulfuric acid, etc.); and conventional catalytic reduction in the presence of a conventional metallic catalyst such as palladium catalysts (e.g. spongy palladium, palladium black, palladium oxide, palladium on carbon, colloidal palladium, palladium on barium sulfate, palladium on barium carbonate, palladium hydroxide on carbon, etc.), nickel catalysts (e.g. reduced nickel, nickel oxide, Raney nickel, etc.), platinum catalysts (e.g. platinum plate, spongy platinum, platinum black, colloidal platinum, platinum oxide, platinum wire, etc.), and the like.

In case that the catalytic reduction is applied, the reaction is preferably carried out around neutral

condition.

This reaction is usually carried out in a

conventional solvent which does not adversely influence the reaction such as water, alcohol (e.g. methanol, ethanol, propanol, etc.), dioxane, tetrahydrofuran, acetic acid, buffer solution (e.g. phosphate buffer, acetate buffer, etc.), and the like, or a mixture thereof.

The reaction temperature is not critical and the reaction is usually carried out under from cooling to warming.

In case that the carboxy-protective group is allyl group, it can be deproteσted by hydrogenolysis using a palladium compound.

Suitable palladium compound used in this reaction may be palladium on carbon, palladium hydroxide on carbon, palladium chloride, a palladium-ligand complex such as tetrakis ( triphenylphosphine)palladium(0),

bis(dibenzylideneacetone)palladium(0), di[1,2-bis(diphenyl phosphino)ethane]palladium(0), tetrakis(triphenyl

phesphite)palladium(0), tetrakis(triethyl phosphite)-palladium(0), and the like.

The reaction can preferably be carried out in the presence of a scavenger of allyl group generated in situ, such as amine (e.g. morpholine, N-methylaniline, etc.), an activated methylene compound (e.g. dimedone, benzoyl acetate, 2-methyl-3-oxovaleric acid, etc.), a cyanohydrin compound (e.g. α-tetrahydropyranyloxybenzyl cyanide, etc.), lower alkanoic acid or a salt thereof (e.g. formic acid, acetic acid, ammonium formate, sodium acetate, etc.), N-hydroxysuccinimide, and the like.

This reaction can be carried out in the presence of a base such as lower alkylamine (e.g. butylamine,

triethylamine, etc.), pyridine, and the like.

When palladium-ligand complex is used in this

reaction, the reaction can preferably be carried out in the presence of the corresponding ligand (e.g.

triphenylphosphine, triphenyl phosphite, triethyl

phoshite, etc.).

This reaction is usually carried out in a

conventional solvent which does not adversely influence the reaction such as water, methanol, ethanol, propanol, dioxane, tetrahydrofuran, acetonitrile, chloroform, dichloromethane, dichloroethane, ethyl acetate, etc., or a mixture thereof.

The removal reaction can be selected according to the kind of carboxy-protective group to be removed.

(3) Process 3 :

The compound (I-d) or salts thereof can be prepared by subjecting the compound (I-c) or salts thereof to removal reaction of the hydroxy-protective group.

Suitable salts of the compounds (I-c) and (I-d) may be the same as those for the compound (I).

This reaction is usually carried out by a

conventional method such as hydrolysis, reduction and the like.

The method of hydrolysis and reduction, and the reaction conditions (e.g. reaction temperature, solvent, etc.) are substantially the same as those illustrated for removal reaction of the carboxy-protective group of the compound (I-a) in Process 2, and therefore are to be referred to said explanation.

In case that the hydroxy-protective group is

tri(lower)alkylsilyl, the removal of this protective group can also be carried out in the presence of

tetra(lower)alkylammonium fluoride (e.g.

tetrabutylammonium fluoride, etc.). (4) Process 4 :

The compound (I-f) or salts thereof can be prepared by subjecting the compound (I-e) or salts thereof to removal reaction of the imino-protective group.

Suitable salts of the compound (I-e) may be salts with bases such as those given for the compound (I). Suitable salts of the compound (I-f) may be the same as those for the compound (I).

This reaction is usually carried out by a

conventional method such as hydrolysis, reduction, and the like.

(5) Process 5 :

The compound (I-g) or salts thereof can be prepared by reacting the compound (I-f) or salts thereof with the compound (III) or salts thereof.

Suitable salts of the compound (I-g) may be the same as those for the compound (I) .

Suitable salts of the compound (III) may be the same acid addition salts as mentioned for the compound (I).

This reaction is usually carried out in a

conventional solvent which does not adversely influence the reaction such as tetrahydrofuran, dioxane, water, methanol, ethanol, buffer solution (e.g. phosphate buffer, etc.), etc., or a mixture thereof.

This reaction can be carried out in the presence of an organic of inorganic base such as those given in the explanation of Process 2.

The reaction temperature is not sritcal and the reaction is usually carried out under from cooling to warming. ( 6 ) Process 6 :

The compound (l-h) or salts thereof can be prepared by reacting the compound (Xl-a) or salts thereof with the compound (Ill-a) or salts thereof.

Suitable salts of the compound (Xl-a) may be the same as those for the compound (I).

Suitable salts of the compound (III-a) may be the same as those for the compound (III).

The method and the reaction conditions (e.g. reaction temperature, solvent, etc.) are substantially the same as those for the Process 5, and therefore are to be referred to said explanation.

(7) Process 7 :

The compound (I-i) or salts thereof can be prepared by reacting the compound (Xl-b) or salts thereof with the compound (III-a) or salts thereof.

Suitable salts of the compound (Xl-b) may be the same as those for the compound (I). The method and the reaction conditions (e.g. reaction temperature, solvent, etc.) are substantially the same as those for the Process 5, and therefore are to be referred to said explanation. (8) Process 8 :

The compound (I-e) or salts thereof can be prepared by introducing the imino-protective group into the

compound (I-f) or salts thereof. Suitable introducing agent of the imino-protective group used in this reaction may be a conventional

acylating agent which is capable of introducing the acyl group as mentioned before such as carboxylic acid, carbonic acid, sulfonic acid and their reactive

derivative, for example, an acid halide, an acid

anhydride, an activated amide, an activated ester, and the like. Preferable example of such reactive derivative may include acid chloride, acid bromide, a mixed acid

anhydride with an acid such as substituted phosphoric acid (e.g. dialkylphosphoric acid, phenylphosphoric acid, diphenylphosphoric acid, dibenzylphosphoric acid,

halogenated phosphoric acid, etc.), dialkylphosphorous acid, sulfurous acid, thiosulfuric acid, sulfuric acid, alkyl carbonate (e.g. methyl carbonate, ethyl carbonate, propyl carbonate, etc.), aliphatic carboxylic acid (e.g. pivalic acid, pentanoic acid, isopentanoic acid,

2-ethylbutyric acid, trichloroacetic acid, etc.), aromatic carboxylic acid (e.g. benzoic acid, etc.), a symmetrical acid anhydride, an activated acid amide with a

heterocyclic compound containing imino function such as imidazole, 4-substituted imidazole, dimethylpyrazole, triazole and tetrazole, an activated ester (e.g.

p-nitrophenyl ester, 2,4-dinitrophenyl ester,

trichlorophenyl ester, pentachlorophenyl ester,

mesylphenyl ester, phenylazophenyl ester, phenyl thioeser, p-nitrophenyl thioester, p-cresyl thioester, carboxymethyl thioester, pyridyl ester, piperidinyl ester, 8-quinolyl thioester, or an ester with a N-hydroxy compound such as N,N-dimethylhydroxylamine, 1-hydroxy-2-(1H)-pyridone, N-hydroxysuccinimide, N-hydroxyphthalimide,

1-hydroxybenzotriazole, 1-hydroxy-6-chlorobenzotriazole, etc.), and the like.

This reaction can be carried out in the presence of an organic or inorganic base such as alkali metal (e.g. lithium, sodium, potassium, etc.), alkaline earth metal (e.g. calcium, etc.), alkali metal hydride (e.g. sodium hydride, etc.), alkaline earth metal hydride (e.g. calcium hydride, etc.), alkali metal hydroxide (e.g. sodium hydroxide, potassium hydroxide, etc.), alkali metal carbonate (e.g. sodium carbonate, potassium carbonate, etc.), alkali metal bicarbonate (e.g. sodium bicarbonate, potassium bicarbonate, etc.), alkali metal alkoxide (e.g. sodium methoxide, sodium ethoxide, potassium tert-butoxide, etc.), alkali metal alkanoic acid (e.g. sodium acetate, etc.), trialkylamine (e.g. triethylamine, etc.), pyridine compound (e.g. pyridine, lutidine, picoline, 4-dimethylaminopyridine, etc.), guinoline, and the like. In case that the introducing agent of the imino- protective group is used in a free form or its salt in this reaction, the reaction is preferably carried out in the presence of a condensing agent such as a carbodiimide compound [e.g. N,N'-dicyclohexylcarbodiimide,

N-cyclohexyl-N'-(4-diethylaminocyclohexyl)carbodiimide, N,N'-diethylcarbodiimide, N,N'-diisopropylcarbodiimide, N-ethyl-N'-(3-dimethylaminopropyl)carbodiimide, etc.], a ketenimine compound (e.g. N,N' -carbonylbis(2- methylimidazole), pentamethyleneketene-N-cyclohexylimine, diphenylketene-N-cyclohexylimine, etc.); an olefinic or acetylenic ether compounds (e.g. ethoxyacetylene,

β-chlorovinylethyl ether), a sulfonic acid ester of

N-hydroxybenzotriazole derivative [e.g. 1-(4-chlorobenzenesulfonyloxy)-6-chloro-1H-benzotriazole, etc.], a combination of trialkylphosphite or triphenylphosphine and carbon tetrachloride, disulfide or

diazenedicarboxylate (e.g. diethyl diazenedicarboxylate, etc.), a phosphorus compound (e.g. ethyl polyphosphate, isopropyl polyphosphate, phosphoryl chloride, phosphorus tirchloride, etc.), thionyl chloride, oxalyl chloride, N-ethylbenzisoxazolium salt, N-ethyl-5-phenylisoxazolium-3-sulfonate, a reagent (referred to a so-called "Vilsmeier reagent") formed by the reaction of an amide compound such as N,N-di ( lower)alkylformamide (e.g. dimethylformamide, etc. ), N-methylformamide or the like with a halogen compound such as thionyl chloride, phosphoryl chloride, phosgene or the like.

This reaction is usually carried out in a

conventional solvent which does not adversely influence the reaction such as water, acetone, dichloromethane, alcohol (e.g. methanol, ethanol, etc.), tetrahydrofuran, pyridine, N,N-dimethylformamide, etc., or a mixture thereof, and further in case that the imino-introducing agent is in liquid, it can also be used as a solvent.

(9) Process 9 :

The compound (I-j) or salts thereof can be prepared by reacting the compound (I-f) or salts thereof with an alkylating agent.

Suitable salts of the compound (I-j) may be the same as those for the compound (I).

Suitable alkylating agent used in this reaction may include a conventional one which is capable of alkylating a hydroxy group to an alkoxy group such as dialkyl sulfate (e.g. dimethyl sulfate, diethyl sulfate, etc.), alkyl sulfonate (e.g. methyl sulfonate, etc.), alkyl halide (e.g. methyl iodide, ethyl iodide, propyl bromide, etc.), diazoalkanes (e.g. diazomethane, diazoethane, etc.), and the like.

This reaction is preferably carried out in the presence of an inorganic or organic base such as those given in the explanation of the Process 2.

Further, this reaction is usually carried out in a conventional solvent which does not adversely influence the reaction such as water, acetone, dichloromethane, methanol, ethanol, propanol, pyridine,

N,N-dimethylformamide, or a mixture thereof.

(10) Process 10 :

The compound (I-k) or salts thereof can be prepared by reacting the compound (XI-c) or salts thereof with the compound (Ill-a) or salts thereof.

Suitable salts of the compound (XI-c) may be the same as those for the compound (XI-a).

Suitable salts of the compound (I-k) may be the same as those for the compound (I).

The method and the reaction conditions (e.g. reaction temperature, solvent, etc.) are substantially the same as •those for the Process 6, and therefore are to be referred to said explanation. Methods for preparing the new starting compounds (II) and (IV-a) or salts thereof are explained in detail in the following.

(A) Method A :

The compound (VII) or salts thereof can be prepared by reacting the compound (V) with the compound (VI) or salts thereof.

Suitable salts of the compound (VI) and (VII) may be the same acid addition salts as those for the compound (I). The compound (VI) or salts thereof can be prepared from the known compounds by a conventional manner or that described in the Preparations of the present

specification.

This reaction can be carried out in the presence of a base such as inorganic base, for example, alkali metal hydroxide (e.g. sodium hydroxide, potassium hydroxide, etc.), alkaline earth metal hydroxide (e.g. magnesium hydroxide, calcium hydroxide, etc.), alkali metal hydride (e.g. sodium hydride, potassium hydride, etc.), alkaline earth metal hydride (e.g. calcium hydride, etc.), alkali metal alkoxide (e.g. sodium methoxide, sodium ethoxide, potassium t-butoxide, etc.), alkali metal carbonate (e.g. sodium carbonate, potassium carbonate, etc.), alkaline earth metal carbonate (e.g. magnesium carbonate, calcium carbonate, etc.), alkali metal bicarbonate (e.g. sodium bicarbonate, potassium bicarbonate, etc.); an organic base, for example, an organic amine (e.g. triethylamine, pyridine, picoline, ethanolamine, triethanolamine,

dicyclohexylamine, N,N'-dibenzylethylenediamine, etc.); and the like.

This reaction can preferably be carried out in the presence of an enolizating agent. Suitable enolizating agent may include tri(lower)alkylsilyl trihalo(lower)-alkanesulfonate, preferably tri(C₁-C₄)alkylsilyl trihalo(C₁-C₄)alkanesulfonate (e.g. trimethylsilyl trifluoro¬methanesulfonate, etc.), tin compound such as stannous ( lower)alkylsulfonate which may have halogen(s),

preferably stannous polyhalo(C₁-C₄)alkylsulfonate (e.g. stannous trifluoromethanesulfonate, etc.), and the like.

This reaction is usually carried out in a conventional solvent which does not adversely influence the reaction such as water, dichloromethane, alcohol (e.g. methanol, ethanol, etc.), tetrahydrofuran, dioxane, acetone, etc., or a mixture thereof. A liquid base can be also used as the solvent.

This reaction temperature is not critical and the reaction is usually carried out under from cooling to warming. (B) Method B :

The compound (II) or salts thereof can be prepared by reacting the compound (VII) or salts thereof with the compound (VIII).

This reaction is usually carried out in a

conventional solvent which does not adversely influence the reaction such as water, dichloromethane, alcohol (e.g. methanol, ethanol, etc.), tetrahydrofuran, dioxane, acetone, benzene, toluene, xylene, etc., or a mixture thereof.

The reaction temperature is not critical and the reaction is usually carried out under from warming to heating.

(C) Method C :

The compound (X) or salts thereof can be prepared by reacting the compound (V) with the compound (IX).

Suitable salts of the compound (X) may be the same as those for the compound (VII).

The method and the reaction conditions (e.g. reaction temperature, solvent, etc.) are substantially the same as those for the Method A, and therefore are to be referred to said explanation.

(D) Method D :

The compound (IV) or salts thereof can be prepared by reacting the compound (X) or salts thereof with the compound (VIII).

Suitable salts of the compound (IV) may be the same as those for the compound (IV-a).

The method and the reaction conditions (e.g. reaction temperature, solvent, etc.) are substantially the same as those for the Method B, and therefore are to be referred to said explanation. (E) Method E :

The compound (IV-a) or salts thereof can be prepared by subjecting the compound (IV) or salts thereof to a removal reaction of the amino-protective group(s).

Suitable salts of the compound (IV-a) may be the same as those for the compound (IV).

The method and the reaction conditions (e.g. reaction temperature, solvent, etc.) are substantially the same as those for Process 4, and therefore are to be referred to said explanation.

(F) Method F :

The compound (XI) or salts thereof can be prepared by cyclizing the compound (IV-a) or salts thereof.

The method and the reaction conditions (e.g. reaction temperature, solvent, etc.) are substantially the same as those for Process 1, and therefore are to be referred to said explanation.

The object compounds obtained according to the above Processes can be isolated and purified in a conventional manner, for example, extraction, precipitation, fractional crystallization, recrystallization, chromatography, and the like.

The object compound (I) and pharmaceutically

acceptable salts thereof of the present invention are novel and exhibit high antimicrobial activity, inhibiting the growth of a wide variety of pathogenic microorganisms including Gram-positive and Gram-negative microorganisms, and further, are very stable against dehydropeptidase and show high urinary excretion, therefore have high potential for the treatment of various infectious diseases.

Now in order to show the utility of the object compound (I) , the test data on antimicrobial activity of the representative compound of the compound (I) of this invention is shown in the following. in vitro Antimicrobial Activity

Test Method : in vitro Antimicrobial Activity was determined by the two-fold agar-plate dilution method as described below.

One loopful of an overnight culture of a test strain in Trypticase-soy broth (10⁶ viable cells per ml) was streaked on heart infusion agar (Hl-agar) containing graded concentrations of the test compound, and the minimal inhibitory concentration (MIC) was expressed in terms of μg/ml after incubation at 37ºC for 20 hours.

Test Compound :

The compound of Example 22

Test Result :

Test Strain MIC (μg/ml)

S. epidermidis 89 ≤ 0.025

For therapeutic administration, the object compound (I) and the pharmaceutically acceptable salts thereof of the present invention are used in the form of conventional pharmaceutical preparation which contains said compound, as an active ingredient, in admixture with

pharmaceutically acceptable carriers such as an organic or inorganic solid or liquid excipient which is suitable for oral, parenteral and external administration. The

pharmaceutical preparations may be in solid form such as tablet, granule, powder, capsule, or liquid form such as solution, suspension, syrup, emulsion, lemonade, and the like.

if needed, there may be included in the above

preparations auxiliary substances, stabilizing agents, wetting agents and other commonly used additives such as lactose, stearic acid, magnesium stearate, terra alba, sucrose, corn starch, talc, gelatin, agar, pectin, peanut oil, olive oil, cacao butter, ethylene glycol, tartaric acid, citric acid, fumaric acid, and the like.

While the dosage of the compound (I) may vary from and also depended upon the age, conditions of the patient, a kind of diseases, a kind of the compound (I) to be applied, etc. In general, amount between 1 mg and about 4,000 mg or even more per day may be administered to a patient. An average single dose of about 1 mg, 10 mg, 50 mg, 100 mg, 250 mg, 500 mg, 1000 mg, 2000 mg of the object compound (I) of the present invention may be used in treating diseases infected by pathogenic microorganisms.

The following Preparations and Examples are given for the purpose of illustrating this invention in more detail. Preparation 1-1)

To the suspension of L-aspartic acid (781.4 g) in a mixture of tetrahydrofuran (4 ℓ) and water (4 ℓ) was added dropwise at 20ºC a solution of allyl chloroformate (685 ml) in tetrahydrofuran (0.3 ℓ) while adjusting pH to around 10 with aqueous sodium hydroxide. After stirring at 10°C for 30 minutes, to the mixture was added sodium chloride and the aqueous layer was separated. After adjusting pH to around 1.5 with hydrochloric acid, the mixture was extracted with ethyl acetate (6 ℓ). The organic layer was washed with brine, dried over magnesium sulfate. Evaporation of the solvent gave

N-allyloxycarbonyl-L-aspartic acid (1690 g).

NMR (DMSO-d₆, δ) : 2.48-2.78 (2H, m) , 3.38 (1H,

br s), 4.25-4.38 (1H, m) , 4.38-4.49 (2H, m) ,

5.10-5.34 (2H, m) , 5.79-6.00 (1H, m) , 7.52 (1H, d, J=8Hz), 12.6 (2H, br s)

Preparation 1-2)

To a solution of N-allyloxycarbonyl-L-aspartic acid

(2.0 g) in ethanol (20 ml) was added sulfuric acid (0.15ml) and the mixture was heated to 100ºC for 1.5 hours.

After cooling to ambient temperature, to the mixture was added triethylamine (0.9 ml) and evaporated. The residue was taken up into ethyl acetate (20 ml), washed in turn with hydrochloric acid (IN), aqueous sodium hydrogen carbonate and brine, and dried over magnesium sulfate.

Evaporation of the solvent gave diethyl

N-allyloxycarbonyl-L-aspartate (2.28 g).

IR (Neat) : 3350, 1710 cm^-1

NMR (CDCl₃, δ) : 1.25 (3H, t, J=4Hz), 1.27 (3H, t, J=4Hz), 2.76-3.08 (2H, m) , 4.09-4.28 (4H, m) , 4.57-4.65 (3H, m) , 5.18-5.38 (2H, m) , 5.83 (1H, m), 5.88-6.00 (1H, m) Preparation 1-3)

To a solution of diethyl N-allyloxycarbonyl-L-aspartatε (100 g) in ethanol (0.5 ℓ) was added by portions sodium borohydride (31.9 g) at 2ºC. After stirring at 2ºC for 3.5 hours, the mixture was diluted with ethyl acetate (1.8 ℓ) and washed with brine (2.5 ℓ, 0.6 ℓ × 2 ) , and dried over magnesium sulfate. Evaporation of the solvent gave a residue which was chromatographed on silica gel (0.5 ℓ) eluting with a mixture of n-hexane and ethyl acetate (1:9-6:4 V/V) to give ethyl

(3S)-3-allyloxycarbonylamino-4-hydroxybutanoate (50.5 g) .

IR (Neat) : 3320, 1710 cm^-1

NMR (CDCl₃, δ) : 1.26 (3H, t, J=6Hz),

2.63 (2H, d, J=5Hz), 2.78 (1H, br s), 3.60-3.80 (2H, m) , 3.80-4.30 (3H, m) ,

4.35-4.68 (2H, m) , 5.05-5.42 (2H, m) , 5.42-5.65 (1H, m) , 5.65-6.15 (1H, m)

Preparation 1-4)

To a solution of ethyl (3S)-3-allyloxycarbonylamino-4-hydroxybutanoate (12.8 g) in dichloromethane (130 ml) were added triethylamine (8.50 ml) and methanesulfonyl chloride (4.50 ml) at -20ºC. After stirring at 0ºC for 10 minutes, the reaction mixture was diluted with ethyl acetate (500 ml), washed in turn with hydrochloric acid (IN), aqueous sodium hydrogen carbonate and brine, and dried over magnesium sulfate. Evaporation of the solvent gave ethyl ( 3S)-3-allyloxycarbonylamino-4-methanesulfonyloxybutanoate (17.12 g).

NMR (CDCl₃, δ) : 1.27 (3H, t, J=6Hz),

2.67 (2H, d, J=6Hz), 3.03 (3H, s),

4.05-4.44 (5H, m) , 4.44-4.62 (2H, m), 5.15-5.60 (3H, m), 5.78-6.02 (1H, m) Preparation 1-5)

To a solution of ethyl (3S)-3-allyloxycarbonylamino- 4-methanesulfonyloxybutanoate (17.1 g) in N,N-dimethylformamide (100 ml) were added ammonium chloride (4.42 g) and sodium azide (5.38 g). After heating at 80ºC for 2.5 hours, the reaction mixture was diluted with ethyl acetate (400 ml), washed in turn with water (200 ml × 4) and brine, and dried over magnesium sulfate. Evaporation of the solvent gave ethyl (3S)-3-allyloxycarbonylamino-4-azidobutanoate (15.47 g).

IR (Neat) : 3320, 2100, 1720 cm^-1

Preparation 1-6)

To a solution of ethyl (3S)-3-allyloxycarbonylamino-4-azidobutanoate (15.4 g) in tetrahydrofuran (70 ml) was added triphenylphosphine (17.34 g). After stirring at ambient temperature for 5 hours, to the mixture was addedaqueous ammonium hydroxide (28%, 7.5 ml). After stirring at ambient temperature for 12 hours, the resulting mixture was evaporated and the residue was taken up into a mixture of tetrahydrofuran (100 ml) and water (100 ml). To the mixture was added dropwise allyl chloroformate (6.4 ml) at 5ºC while adjusting pH to around 10 with aqueous sodium carbonate. The reaction mixture was diluted with ethyl acetate (300 ml), washed in turn with water, hydrochloric acid (1N), aqueous sodium hydrogen carbonate and brine, and dried over magnesium sulfate. Evaporation gave a residue, which was chromatographed on silica gel (150 ml) eluting with a mixture of n-hexane and ethyl acetate

(5:95-30:70 V/V) to give ethyl (3S)-3,4-bis(allyloxycarbonylamino)butanoate (5.13 g).

IR (Nujol) : 3310, 1730, 1680 cm^-1

NMR (CDCl₃, δ) : 1.27 (3H, t, j=7Hz), 2.59 ( 2H, d, J=6Hz), 3.39 (2H, t, J=6Hz), 3.85-4.33 (3H, m), 4.35-4.70 (4H, m) , 5.00-6.20 (8H, m) Preparation 1-7)

To a solution of ethyl (3S)-3,4-bis- (allyloxycarbonylamino)butanoate (57.87 g) in ethanol (460 ml) was added aqueous potassium hydroxide (2N, 110 ml). After stirring at ambient temperature for 3 hours, to the mixture was added acetic acid (12.0 ml) and the mixture was diluted with ethyl acetate (1 ℓ) and water (0.8 ℓ) . The aqueous layer was separated, cooled to 10°C and diluted with ethyl acetate (1 ℓ). After adjusting pH to around 1.5 with aqueous hydrochloric acid (6N), the organic layer was separated, washed with brine, and dried over magnesium sulfate. Evaporation of the solvent gave (3S)-3,4-bis(allyloxycarbonylamino)butanoic acid (63.97 g).

NMR (DMSO-d₆, δ) : 2.05-2.30 (2H, m) , 2.85-3.22 (2H, m), 3.68-4.05 (1H, m) , 4.25-4.60 (4H, m),

5.00-5.20 (4H, m), 5.78-6.02 (2H, m), 7.00-7.20 (2H, m), 12.1 (1H, br s) Preparation 1-8)

To a suspension of (3S)-3,4-bis-(allyloxycarbonylamino)butanoic acid (63.95 g) in

dichloromethane (750 ml) were added successively Meldrum's acid (27.85 g) , dicyclohexylcarbodiimide (39.86 g) and 4-dimethylaminopyridine (25.18 g) at 5ºC. After stirring at ambient temperature for 12 hours, the mixture was washed in turn with hydrochloric acid (1N, 220 ml), cold water (200 ml) and brine (200 ml), and evaporated. The residue was taken up into a mixture of acetic acid (250 ml) and water (250 ml) and the resulting mixture was heated to reflux for 1 hour. Evaporation of the solvent gave a residue which was dissolved in ethyl acetate and washed successively with aqueous sodium hydrogen carbonate and brine, and dried over magnesium sulfate. Evaporation of the solvent gave a residue which was taken up into diisopropyl ether. The resulting presicipitate was collected by filtration, dried in vacuo to give

(4S)-4,5-bis(allyloxycarbonylamino)ρentan-2-one (31.0 g).

IR (Nujol) : 3310, 1710, 1680 cm^-1

NMR (CDCl₃, δ) : 2.16 (3H, s), 2.45-2.87 (2H, m),

3.30-3.45 (2H, m) , 3.95-4.10 (1H, m), 4.50-4.60 (4H, m) , 5.10-5.36 (5H, m), 5.50-5.80 (1H, m), 5.80-6.02 (2H, m) Preparation 1-9)

To a solution of (4S)-4,5-bis(allyloxycarbonylamino)¬pentan-2-one (29.67 g) in dichloromethane (300 ml) were added N-ethylpiperidine (43.0 ml) and trimethylsilyl trifluoromethanesulfonate (60.5 ml) at -10°C and the resulting mixture was allowed to stir at ambient

temperature for 2.5 hours. To a solution of

(3R,4R)-4-acetoxy-3-[(1R)-1-t-butyldimethylsilyloxyethyl]-2-oxoazetidine (44.99 g) in dichloromethane (270 ml) were added N-ethylpiperidine (21.5 ml) and trimethylsilyl trifluoromethanesulfonate (30.3 ml) at -10ºC and the resulting mixture was allowed to stir at 0ºC for 1 hour. To a mixture of the latter solution and zinc bromide (94.4 g) was added the former solution at 0°C and the resulting mixture was allowed to stir at ambient temperature for 3 hours. The reaction mixture was diluted with a mixture of ethyl acetate (1.5 ℓ) and water (1.5 ℓ). After adjusting pH to 1 with hydrochloric acid (6N). The mixture was allowed to stir at ambient temperature for additional 1 hour. The separated organic layer was washed with water and diluted with water (1.5 ℓ). After adjusting pH to 7 with aqueous sodium hydrogen carbonate, the organic layer was washed with brine and dried over magnesium sulfate. Evaporation of the solvent gave a residue, which was chromatographed on silica gel (1.8 ℓ) eluting with a mixture of n-hexane and ethyl acetate (1:9-9:1 V/V) to give (3S,4R)-4-[(4S)-4,5-bis(allyloxycarbonylamino)-2- oxopentyl]-3-[(1R)-1-t-butyldimethylsilyloxyethyl]-2-oxoazetidine (17.12 g).

IR (CH₂Cl₂) : 3430, 1760, 1720 cm^-1

NMR (CDCl₃, δ) : 0.07 (6H, s), 0.87 (9H, s),

1.21 (3H, d, J=6Hz), 2.50-3.10 (5H, m),

3.20-3.41 (2H, m) , 3.90-4.21 (3H, m), 4.49-4.57 (4H, m) , 5.10-5.40 (5H, m), 5.60-5.80 (1H, m), 5.80-6.00 (2H, m), 6.39 (1H, br s)

Preparation 1-10)

To a solution of (3S,4R)-4-[(4S)-4,5-bis(allyloxycarbonylamino)-2-oxopentyl]-3-[(1R)-1-t-butyldimethylsilyloxyethyl]-2-oxoazetidine (16.88 g) and triethylamine (9.2 ml) in dichloromethane (140 ml) was added allyl oxalyl chloride (6.5 ml) at -20ºC and the resulting mixture was stirred for 30 minutes at -20ºC. The reaction mixture was taken up into a mixture of ethyl acetate (500 ml) and water (500 ml). The organic layer was separated, washed in turn with water, aqueous sodium hydrogen

carbonate and brine, and dried over magnesium sulfate. Evaporation of the solvent gave a residue which was dissolved in triethyl phosphite (19.8 ml). The mixture was heated at.90ºC for 2 hours under nitrogen atmosphere, and to this mixture was added hydroquinone (5.45 g) and toluene (170 ml), and the resulting mixture was heated to reflux for additional 4.5 hours. After cooling to ambient temperature, the reaction mixture was washed in turn with aqueous sodium carbonate (10%, 80 ml × 5), cold water (80 ml × 2) and brine (80 ml × 2), and dried over magnesium sulfate. Evaporation of the solvent gave a residue, which was chromatographed on silica gel (320 ml) eluting with a mixture of n-hexane and ethyl acetate (1:9-3:7 V/V) to give allyl (5R,6S)-3-[(2S)-2,3-bis(allyloxycarbonylamino)- propyl]-6-[(1R)-1-t-butyldimethylsilyloxyethyl]-7-oxo-1-azabicyclo[3.2.0]hept-2-ene-2-carboxylate (19.56 g).

IR (CH₂Cl₂) : 3430, 1770, 1720 cm^-1

NMR (CDCl₃, δ) : 0.08 (6H, s), 0.90 (9H, s),

1.20 (3H, d, J=6Hz), 1.80-4.35 (10H, m),

4.35-4.82 (6H, m) , 4.95-5.60 (8H, m), 5.60-6.15 (3H, m)

Preparation 1-11)

To a solution of allyl (5R,6S)-3-[(2S)-2,3- bis(allyloxycarbonylamino)propyl]-6-[(1R)-1-t-butyldimethylsilyloxyethyl]-7-oxo-1-azabicyclo[3.2.0]hept-2-ene-2-carboxylate (19.56 g) and acetic acid (3.97 ml) in tetrahydrofuran (70 ml) was added aqueous

tetrabutylammonium fluoride (70%, 25.9 g). After standing at ambient temperature for 15 hours, the reaction mixture was diluted with ethyl acetate (200 ml), washed

successively with water (200 ml × 4) and brine, and dried over magnesium sulfate. Evaporation of the solvent gave a residue, which was chromatographed on silica gel (300 ml) eluting with a mixture of n-hexane and ethyl acetate

(2:8-10:0 V/V) to give allyl ( 5R,6S)-3-[(2S)-2,3-bis(allyloxycarbonylamino)propyl]-6-[(IR)-1-hydroxyethyl]-7-oxo-l-azabicyclo[3.2.0]hept-2-ene-2-carboxylate(4.52 g).

IR (CH₂Cl₂) : 3600, 3420, 1770, 1720 cm^-1

NMR (CDCl₃, δ) : 1.30 (3H, d, J=6Hz),

2.00-4.15 (11H, m), 4.18-4.82 (6H, m), 5.10-5.55 (8H, m) , 5.85-6.10 (3H, m) Preparation 2-1)

N-Allyloxycarbonyl-D-aspartic acid was obtained in 82.0% yield in substantially the same manner as that of Preparation 1-1).

NMR (DMSO-dg, δ) : 2.48-2.78 (2H, m) , 3.38 (1H,

br s), 4.25-4.38 (1H, m) , 4.38-4.49 (2H, m) , 5.10-5.34 (2H, m) , 5.79-6.00 (1H, m),

7.52 (1H, d, J=8Hz), 12.6 (2H, br s)

Preparation 2-2)

Diethyl N-allyloxycarbonyl-D-aspartate was obtained in 89.1% yield in substantially the same m-αnner as that of Preparation 1-2).

IR (Neat) : 3350, 1710 cm^-1

NMR (CDCl₃, δ) : 1.25 (3H, t, J=4Hz),

1.27 (3H, t, J=4Hz), 2.76-3.08 (2H, m),

4.09-4.28 (4H, m), 4.57-4.65 (3H, m), 5.18-5.38 (2H, m), 5.83 (1H, m),

5.88-6.00 (1H, m) Preparation 2-3)

Ethyl (3R) -3-allyloxycarbonylamino-4-hydroxybutanoate was obtained in 45.8% yield in substantially the same manner as that of Preparation 1-3).

IR (Neat) : 3320, 1710 cm^-1

NMR (CDCl₃, δ) : 1.26 (3H, t, J=6Hz),

2.63 (2H, d, J=5Hz), 2.78 (1H, br s), 3.60-3.80 (2H, m), 3.80-4.30 (3H, m), 4.35-4.68 (2H, m), 5.05-5.42 (2H, m), 5.42-5.65 (1H, m), 5.65-6.15 (1H, m)

Preparation 2-4)

Ethyl (3R)-3-allyloxycarbonylamino-4-methanesulfonyl-oxybutanoate was obtained quantitatively in substantially the same manner as that of Preparation 1-4).

NMR (CDCl₃, δ) : 1.27 (3H, t, J=6Hz),

2.67 (2H, d, J=6Hz), 3.03 (3H, s),

4.05-4.44 (5H, m) , 4.44 -4.62 (2H, m) , 5.15-5.60 (3H, m) , 5.78-6.02 (1H, m) Preparation 2-5)

Ethyl (3R)-3-allyloxycarbonylamino-4-azidobutanoate was obtained in substantially the same manner as that of Preparation 1-5).

IR (Neat) : 3320, 2100, 1720 cm^-1

Preparation 2-6)

Ethyl (3R)-3,4-bis(allyloxycarbonylamino)butanoate was obtained in 55.9% yield (calculated based on ethyl (3R)-3-allyloxycarbonylamino-4-hydroxybutanoate) in substantially the same manner as that of Preparation 1-6).

IR (Nujol) : 3310, 1730, 1680 cm^-1

NMR (CDCl₃, δ) : 1.27 (3H, t, J=7Hz), 2.59 (2H, d, J=6Hz), 3.39 (2H, t, J=6Hz), 3.85-4.33 (3H, m), 4.35-4.70 (4H, m) , 5.00-6.20 (8H, m)

Preparation 2-7)

(3R)-3,4-Bis(allyloxycarbonylamino)butanoic acid was obtained in 95.6% yield in substantially the same manner as that of Preparation 1-7).

NMR (DMSO-d₆, δ) : 2.05-2.30 (2H, m), 2.85-3.22 (2H, m), 3.68-4.05 (1H, m) , 4.25-4.60 (4H, m),

5.00-5.20 (4H, m), 5.78-6.02 (2H, m), 7.00-7.20 (2H, m), 12.1 (1H, br s)

Preparation 2-8)

( 4R) -4,5-Bis(allyloxycarbonylamino)pentan-2-rone was obtained in 72.9% yield in substantially the same manner as that of Preparation 1-8).

IR (Nujol) : 3310, 1710, 1680 cm^-1

NMR (CDCl₃, δ) : 2.16 (3H, s), 2.45-2.87 (2H, m) , 3.30-3.45 (2H, m), 3.95-4.10 (1H, m), 4.50-4.60 (4H, m), 5.10-5.36 (5H, m), 5.50-5.80 (1H, m), 5.80-6.02 (2H, m) Preparation 2-9)

(3S,4R)-4-[(4R)-4,5-Bis(allyloxycarbonylamino)-2-oxopentyl]-3-[(1R)-1-t-butyldimethylsilyloxyethyl]-2-oxoazetidine was obtained in 39.4% yield in substantially the same method as that of Preparation 1-9).

IR (CH₂Cl₂) : 3410, 1760, 1720 cm^-1

NMR (CDCl₃, δ) : 0.07 (6H, s), 0.89 (9H, s),

1.20 (3H, d, J=6Hz), 2.50-3.00 (5H, m), 3.35 (2H, t, J=6Hz), 3.90-4.25 (3H, m), 4.50-4.60 (4H, m), 5.10-5.40 (5H, m),

5.50-5.70 (1H, m), 5.80-6.00 (2H, m), 6.28 (1H, br s)

Preparation 2-10)

Allyl (5R,6S)-3-[(2R)-2,3-bis(allyloxycarbonylamino)-propyl]-6-[(1R)-1-t-butyldimethylsilyloxyethyl]-7-oxo-1- azabicyclo[3.2.0]hept-2-ene-2-carboxylate was obtained in 77.9% yield in substantially the same method as that of Preparation 1-10).

IR (CH₂Cl₂) : 3430, 1770, 1720 cm^-1

NMR (CDCl₃, δ) : 0.06 (6H, s), 0.89 (9H, s),

1.22 (3H, d, J=6Hz), 1.70-4.30 (10H, m),

4.30-4.90 (6H, m) , 4.95-5.60 (8H, m)

5.60-6.12 (3H, m)

Preparation 2-11)

Allyl (5R,6S)-3-[(2R)-2,3-bis(allyloxycarbonylamino)-propyl]-6-[(1R)-1-hydroxyethyl]-7-oxo-l-azabicyclo[3.2.0]-hept-2-ene-2-carboxylate was obtained in 56.5% yield in substantially the same method as that of Preparation

1-11).

IR (Nujol) : 3410, 3320, 1760, 1700 cm^-1

NMR (CDCl₃, δ) : 1.32 (3H, d, J=6Hz), 2.00-4.15

(11H, m), 4.15-4.82 (6H, m) , 5.00-5.55 (8H, m) , 5.85-6.10 (3H, m) Preparation 3-1)

To a solution of 1,3-diamino-2-hydroxypropane (10 g) in a mixture of tetrahydrofuran (200 ml) and water (100 ml) were added triethylamine (34 ml) and

di-t-butyldicarbonate (51 g) . After stirring at ambient temperature for 12 hours, the reaction mixture was diluted with ethyl acetate (400 ml) and water (400 ml). The organic layer was washed in turn with hydrochloric acid (1N), aqueous sodium hydrogen carbonate and brine, and dried over magnesium sulfate. Evaporation of the solvent gave a residue, which was taken up into ethyl acetate (350 ml). To the solution were added triethylamine (39 ml) and methanesulfonyl chloride (10.3 ml) at 0ºC. After stirring at 0°C for 30 minutes, the reaction mixture was washed with water, hydrochloric acid (1N), aqueous sodium

hydrogen carbonate and brine, and dried over magnesium sulfate. Evaporation of the solvent gave a crystalline, which was collected by filtration and washed with n-hexane to give 1,3-bis(t-butyloxycarbonylamino)-2-methanesulfonyloxypropane (34 g).

IR (Nujol) : 3460, 3400, 1700 cm^-1

NMR (CDCl₃, δ) : 1.44 (18H, s), 3.09 (3H, s),

3.15-3.60 (4H, m) , 4.60-4.75 (1H, m) , 5.20-5.40 (2H, m)

Preparation 3-2)

To a solution of 1,3-bis(t-butyloxycarbonylamino)-2-methanesulfonyloxypropane (58.4 g) in dimethyl sulfoxide (475 ml) was added sodium cyanide (31 g) and the mixture was heated at 90ºC for 3 hours. After cooling to ambient temperature, the mixture was diluted with ethyl acetate (1.5 ℓ) and water (1.5 ℓ) . The organic layer was washed in turn with water and brine, and dried over magnesium sulfate. Evaporation of the solvent gave 3,4-bis(t-butyloxycarbonylamino)butyronitrile (26.5 g). IR (Nujol) : 3370, 2260, 1680 cm^-1

NMR (CDCl₃, δ) : 1.44 (18H, s), 2.64 (2H, d,

J=6Hz), 3.20-3.44 ( 2H, m) , 3.80-3.98 (1H, m) ,

5.00 (1H, br s), 5.50 (1H, br s)

Preparation 3-3)

To a solution of 3,4-bis(t-butyloxycarbonylamino)-butyronitrile (2.0 g) in methanol (10 ml) was added sulfuric acid (5 ml), and the resulting mixture was heated at 100ºC for 5 hours. After cooling to ambient

temperature, the reaction mixture was poured into a mixture of ice and water (30 g) , and tetrahydrofuran (30 ml) was added thereto. To the solution was added allyl chloroformate (1.5 ml) at 0ºC while adjusting pH to around 9 with aqueous potassium carbonate. The reaction mixture was diluted with ethyl acetate (150 ml), washed with brine, dried over magnesium sulfate. Evaporation of the solvent gave a residue, which.was chromatographed on silica gel (60 ml) eluting with a mixture of n-hexane and ethyl acetate (95:5-6:4 V/v) to give methyl

3,4-bis(allyloxycarbonylamino)butanoate (1.90 g) .

NMR (CDCl₃, δ) : 2.60 (2H, d, J=6Hz), 3.38 (2H, t,

J=6Hz), 3.70 (3H. s), 3.85-4.35 (1H, m) ,

4.40-4.70 (4H, m) , 5.03-5.60 (5H, m) , 5.60-6.20 (3H, m)

Preparation 3-4)

3,4-Bis(allyloxycarbonylamino)butanoic acid was obtained in 96.7% yield in substantially the same method as that of Preparation 1-7).

NMR (DMSO-d₆, δ) : 2.05-2.30 (2H, m) , 2.85-3.22 (2H, m), 3.68-4.05 (1H, m) , 4.25-4.60 (4H, m) ,

5.00-5.20 (4H, m), 5.78-6.02 (2H, m) , 7.00-7.20 (2H, m), 12.1 (1H, br s) Preparation 3-5)

4, 5-Bis(allyloxycarbonylamino)pentan-2-one was obtained in 54.7% yield in substantially the same manner as that of Preparation 1-8).

IR (Nujol) : 3310, 1710, 1680 cm^-1

NMR (CDCl₃, δ) : 2.16 (3H, s), 2.45-2.87 (2H, m) ,

3.30-3.45 (2H, m) , 3.95-4.10 (1H, m) , 4.50-4.60 (4H, m), 5.10-5.36 (5H, m) , 5.50-5.80 (1H, m) , 5.80-6.02 (2H, m)

Preparation 3-6)

( 3S,4R)-4-[4,5-Bis(allyloxycarbonylamino)-2-oxopentyl]-3-[(1R)-1-t-butyldimethylsilyloxyethyl]-2-oxoazetidine was obtained in 31.0% yield in substantially the same method as that of Preparation 1-9).

NMR (CDCl₃, δ) : 0.07 (6H, s), 0.87 (9H, s),

1.21 (3H, d, J=6Hz), 2.40-3.10 (5H, m), 3.17-3.46 (2H, m) , 3.80-4.25 (3H, m), 4.49-4.57 (4H, m) , 5.05-5-40 (5H, m), 5.40-6.20 (3H, m) , 6.40 (1H, br s)

Preparation 3-7)

Allyl (5R,6S)-3-[2,3-bis(allyloxycarbonylamino)-propyl]-6-[(1R)-1-t-butyldimethylsilyloxyethyl]-7-oxo-1-azabicyclo[3.2.0]hept-2-ene-2-carboxylate was obtained quantitatively in substantially the same method as that of Preparation 1-10 ) .

IR (CH₂Cl₂) : 3430, 1770, 1720 cm^-1

NMR (CDCl₃, δ) : 0.06 (6H, s), 0.89 (9H, s),

1.22 (3H, d, J=6Hz), 1.70-4.30 (10H, m),

4.30-4.90 (6H, m) , 4.95-5.60 (8H, m), 5.60-6.12 (3H, m)

Preparation 3-8)

Allyl (5R,6S)-3-[2,3-bis(allyloxycarbonylamino)- propyl-6-[(1R)-1-hydroxyethyl]-7-oxo-1-azabicyclo[3.2.0]-hept-2-ene-2-carboxylate was obtained in 67.1% yield in substantially the same method as that of Preparation 1-11).

NMR (CDCl₃, δ) : 1.30 (3H, d, J=6Hz), 2.00-4.35

(11H, m), 4.35-4.82 (6H, m) , 5.00-5.60 (8H, m), 5.85-6.20 (3H, m)

Preparation 4-1)

N-Allyloxycarbonyl-N-(2-allyloxycarbonylaminoethyl)-aminoacetic acid was obtained in 80.9% yield from

N-(2-aminoethyl)aminoacetic acid (9.33 g) in substantially the same method as that of Preparation 1-1).

IR (CH₂Cl₂) : 3440, 3300-2200, 1710 cm^"1

NMR (CDCl₃, δ) : 3.20-3.65 (4H, m) , 4.11 (2H, s),

4.45-4.75 (4H, m) , 5.10-5.50 (4H, m), 5.60-6.30 (3H, m)

Preparation 4-2)

3-[N-Allyloxycarbonyl-N-(2-allyloxycarbonylamino¬ethyl)amino]propan-2-one was obtained in 81.6% yield in substantially the same method as that of Preparation 1-8).

IR (CH₂Cl₂) : 3430, 1700 cm^-1

NMR (CDCl₃, δ) : 2.17 (3H, s), 3.13-3.57 (4H, m), 4.07 (2H, s), 4.40-4.65 (4H, m) , 5.00-5.60

(5H, m) , 5.60-6.14 (2H, m)

Preparation 4-3)

To a solution of 3-[N-allyloxycarbonyl-N-(2-allyloxy¬carbonylaminoethyl)amino]propan-2-one (11.40 g) in

dichloromethane (100 ml) were added triethylamine (12.3 ml) and trimethylsilyl trifluoromethanesulfonate (15.5 ml) at -10ºC and the resulting mixture was allowed to stir at 0ºC for 1 hour. To a solution of (3R,4R)-4-acetoxy-3-[(1R)-1-t-butyldimethylsilyloxyethyl]-2-oxoazetidine (11.54 g) in dichloromethane (60 ml) were added

triethylamine (5.6 ml) and trimethylsilyl

trifluoromethanesulfonate (7.76 ml) at -10°C and the resulting mixture was allowed to stir at 0°C for 1 hour. To the latter solution were added the former solution and trimethylsilyl trifluoromethanesulfonate (3.8 ml) at 0°C and the resulting mixture was allowed to stir at 0ºC for 2 hours. The reaction mixture was diluted with ethyl acetate (500 ml) and water (500 ml). After adjusting pH to 1 with hydrochloric acid (6N), the mixture was stirred at ambient temperature for 1 hour. After adjusting pH to 7 with aqueous sodium hydrogen carbonate, the organic phase was separated, washed with brine, dried over

magnesium sulfate. Evaporation of the solvent gave a residue which was chromatographed on silica gel (600 ml) eluting with a mixture of n-hexane and ethyl acetate

(7:3-0:10 V/V),to give ( 3S,4R)-4-[3-{N-allyloxycarbonyl-N-(2-allyloxycarbonylaminoethyl)amino}-2-oxopropyl]-3-[(1R)-1-t-butyldimethylsilyloxyethyl]-2-oxoazetidine (11.8 g).

IR (CH₂Cl₂). : 3400, 1760, 1720 cm^-1

NMR (CDCl₃, δ) : 0.09 (6H, s), 0.89 (9H, s),

1.23 (3H, d, J=6Hz), 2.55-2.90 (3H, m), 3.10-3.60 (4H, m), 3.80-4.35 (4H, m), 4.35-4.67 (4H, m) , 5.00-5.50 (5H, m), 5.55-6.25 (2H, m) , 6.30 (1H, br s)

Preparation 4-4)

Allyl ( 5R,6S)-3-[N-allyloxycarbonyl-N-(2-allyloxycarbonylaminoethyl)aminomethyl]-6-[(1R)-1-t-butyldimethylsilyloxyethyl]-7-oxo-1-azabicyclo[3.2.0]-hept-2-ene-2-carboxylate was obtained in 80.2% yield in substantially the same manner as that of Preparation

1-10).

NMR (CDCl₃, δ) : 0.06 (6H, s), 0.87 (9H, s),

1.19 (3H, d, J=6Hz), 2.77 (2H, d, J=9Hz), 3.03 (1H, dd, J=3, 6Hz), 3.17-3.42 (4H, m), 3.88-4.30 (2H, m), 4.30-4.70 (8H, m) , 4.70-5.50 (7H, m), 5.55-6.10 (3H, m) Preparation 4-5)

Allyl (5R,6S)-3-[N-allyloxycarbonyl-N-(2-allyloxycarbonylaminoethyl)aminomethyl]-6-[(1R)-1-hydroxyethyl]-7-oxo-1-azabicyclo[3.2.0]hept-2-ene-2-carboxylate was obtained in 62.1% yield in substantially the same method as that of Preparation 1-11).

NMR (CDCl₃, δ) : 1.35 (3H, d, J=6Hz), 2.89 (2H, d, J=9Hz), 3.18 (1H, dd, J=3 , 6Hz), 3.26-3.50 (4H, m), 3.98-4.40 (2H, m) , 4.40-4.83 (8H, m) ,

4.83-5.60 (7H, m) , 5.66-6.22 (3H, m)

Preparation 5

To a solution of LDA (lithium diisopropylamide) in tetrahydrofuran (0.0135 mole) was added a solution of ( 5S)-8-oxo-2-phenyl-1-aza-3-oxabicyclo[3.3.0]octane (2.5 g) in tetrahydrofuran (50 ml) at -78ºC under nitrogen.

The mixture was stirred for 30 minutes at -78°C, and allyl bromide (1.3 ml) was added to the reaction mixture at -78ºC. The reaction temperature was raised to -20ºC over a period of 30 minutes, and the solution was poured into ice-water, and sodium chloride was added to saturate the aqueous phase. The organic phase was separated and the aqueous phase was extracted with ethyl acetate. The combined organic phase was dried over magnesium sulfate, and concentrated. The residue was chromatographed on silica gel eluting with a mixture of n-hexane and ethyl acetate (4:1 ~ 2:1, V/V) to give (5S,7S)-7-allyl-8-oxo-2-phenyl-1-aza-3-oxabicyclo[3.3.0]octane (1.1 g) .

IR (Neat) : 1700, 1640 cm^-1

NMR (CDCl₃, δ) : 2.0-3.0 (5H, m) , 3.41 (1H, t,

J=8Hz), 3.9-4.4 (2H, m) , 5.0-5.2 (2H, m) , 5 .7-6 . 0 ( 1H, m) , 6 . 32 ( 1H, s ) , 7 . 2-7 . 4 ( 5H, m)

Elution was continued to give ( 5S,7R)-7-allyl-8-oxo-2-phenyl-1-aza-3-oxabicyclo[3.3.0]octane (1.3 g).

IR (Neat) : 1700, 1640 cm^-1

NMR (CDCl₃, δ) : 1.4-1.8 (1H, m) , 2.0-3.0 (5H, m) ,

3.3-3.6 (1H, m), 4.0-4.3 (2H, m) , 5.0-5.2 (2H, m), 5.6-5.9 (1H, m) , 6.32 (1H, s), 7.2-7.5 (5H, m) Preparation 6-1)

To a solution of (5S,7S)-7-allyl-2-phenyl-8-oxo-1-aza-3-oxabicyclo[3.3.0]octane (11.8 g) in dichloromethane (200 ml) was added m-chloroperbenzoic acid (15.7 g) .

After stirring for 12 hours at ambient temperature, the solution was washed in turn with saturated sodium

bicarbonate and brine. The solvent was dried and

evaporated. The residue was chromatographed on silica gel eluting with -a mixture of n-hexane and ethyl acetate (2:1, V/V) to give (5S,7R)-7-(2,3-epoxypropyl)-8-oxo-2-phenyl-1-aza-3-oxabicyclo[3.3.0]octane (8.38 g).

IR (Neat) : 1700 cm^-1

NMR (CDCl₃, δ) : 1.6-3.2 (8H, m) , 3.5-3.7 (1H, m) ,

4.0-4.4 (2H, m), 6.32, 6.34 (1H, each s), 7.2-7.6 (5H, m)

Preparation 6-2)

To a solution of (5S,7R)-7-(2,3-epoxypropyl)-8-oxo-2-phenyl-1-aza-3-oxabicyclo[3.3.0]octane (13.51 g) in tetrahydrofuran (300 ml) was added lithium aluminum

hydride (3.0 g). After stirring for 1 hour under reflux, the solution was cooled to 0ºC and saturated potassium sodium tartrate solution was added thereto. After

stirring for 1 hour, the obtained precipitated material was filtered off. The solution was dried over magnesium sulfate and evaporated to give (2S,4R)-1-benzyl-2- hydroxymethyl-4- ( 2-hydroxypropyl)pyrrolidine ( 14 g) .

IR (Neat) : 3350 cm^-1

NMR ( CDCl₃ , δ ) : 1. 1-1. 3 ( 3H, m) , 3 . 1-4 . 0 ( 7H, m) ,

7. 1-7. 5 ( 5H, m)

Preparation 6-3)

To a solution of (2S,4R)-1-benzyl-2-hydroxymethyl-4-(2-hydroxypropyl)pyrrolidine (14.0 g) in methanol (300 ml) was added 10% palladium on carbon (50% wet, 5g).

After stirring for 2 hours at ambient temperature, the precipitate was filtered off. The solvent was evaporated, and then the residue was dissolved in a mixture of

tetrahydrofuran (100 ml) and water (100 ml). To this solution was added dropwise allyloxycarbonyl chloride (7.1 ml) at 0ºC while keeping pH 9.0-10.0 with 4N-agueous sodium hydroxide. The reaction mixture was extracted with ethyl acetate. The dried solvent was evaporated, and then the residue was chromatographed on silica gel eluting with a mixture of n-hexane and ethyl acetate (1:2, V/V) to give ( 2S, 4R)-1-allyloxycarbonyl-2-hydroxymethyl-4-(2-hydroxy-propyl)pyrrolidine (9.4 g) .

IR (Neat) : 3400, 1630 cm^-1

NMR (CDCl₃, δ) : 1.20, 1.21 (3H, each d, J=6Hz),

1.3-1.9 (3H, m), 2.1-2.4 (2H, m) ,

2.8-3.0 (1H, m) , 3.5-4.1 (5H, m) ,

4.5-4.7 (2H, m), 5.1-5.4 (2H, m),

5.8-6.1 (1H, m)

Preparation 6-4)

To a solution of (2S,4R)-1-allyloxycarbonyl-2-hydroxymethyl-4-(2-hydroxypropyl)ρyrrolidine (6.34 g) in acetone (150 ml) was added 2.6N-Jones reagent (40 ml) at 0ºC. After stirring for 1 hour at 0ºC, the precipitate was filtered off. After the solvent was evaporated, the residue was dissolved in ethyl acetate, and washed with brine. The dried solvent was evaporated to give (2S,4R)- 1-allyloxycarbonyl-2-carboxy-4-(2-oxopropyl)pyrrolidine (7.0 g).

IR (Neat) : 1680-1700 cm^-1

NMR (CDCl₃, δ) : 2.15 (3H, s), 2.4-2.8 (4H, m),

3.0-3.2 (1H, m), 3.8-4.0 (1H, m) , 4.3-4.4 (1H, m), 4.5-4.7 (2H, m) , 5.1-5.4 (2H, m) , 5.7-6.0 (1H, m) Preparation 6-5)

To a solution of (2S,4R)-1-allyloxycarbonyl- 2-carboxy-4-(2-oxopropyl)pyrrolidine (7.0 g) in

dichloromethane (140 ml) were added anhydrous

1-hydroxybenzotriazole (3.9 g), dimethylamine

hydrochloride (2.35 g) and l-ethyl-3-(3- dimethylaminopropyl) carbodiimide (5.3 ml) at 0ºC. After stirring at ambient temperature for 5 hours, the mixture was washed with saturated aqueous sodium bicarbonate and brine, and dried over magnesium sulfate. Evaporation of the solvent gave an oil, which was chromatographed on silica gel eluting with a mixture of dichloromethane and methanol (20:1, V/V) to give (2S,4R)-1-allyloxycarbonyl- 2-(N,N-dimethylcarbamoyl)-4-(2-oxopropyl)pyrrolidine (6.46 g).

IR (Neat) : 1700, 1650 cm^-1

NMR (CDCl₃, δ) : 2.13 (3H, s) , 2.4-2.8 (4H, m),

2.95 (3H, s), 3.10 (3H, s), 3.8-4.0 (1H, m), 4.4-4.7 (3H, m), 5.0-5.5 (2H, m) ,

5.7-6.0 (1H, m)

Preparation 6-6)

(3S,4R)-4-{3-[(2S,4R)-1-Allyloxycarbonyl-2-(N,N-dimethy1carbamoyl)pyrrolidin-4-yl]-2-oxopropyl}-3-[(1R)-1-t-butyldimethylsilyloxyethyl]-2-oxoazetidine was obtained in 66.6% yield in substantially the same manner as that of Preparation 1-9).

IR (CH₉Cl₂) : 1750, 1700, 1650 cm^-1

NMR (CDCl₃, δ) : 0.06 (6H, s), 0.88 (9H, s),

1.19 (3H, d, J=7Hz), 2.95 (3H, s),

3.08 (3H, s), 3.5-3.7 (1H, m) ,

3.8-4.0 (2H, m), 4.4-4.8 (3H, m) ,

5.1-5.4 (2H, m), 5.7-6.0 (1H, m) ,

6.11 (1H, br s) Preparation 7-1)

(5S,7S)-2-Phenyl-7-(2,3-epoxypropyl)-8-oxo-1-aza-3-oxabicyclo[3.3.0]octane was obtained in 99.3% yield in substantially the same manner as that of Preparation 6-1).

IR (Neat) : 1700 cm^-1

NMR (CDCl₃, δ) : 1.4-3.1 (8H, m),

3.43 (1H, dt, J=2, 8Hz), 4.0-4.4 (2H, m),

6.32,6.33 (1H, each s), 7.2-7.5 (5H, m)

Preparation 7-2)

(2S,4S)-1-Benzyl-2-hydroxymethyl-4-(2-hydroxypropyl)-pyrrolidine was obtained quantitatively in substantially the same manner as that of Preparation 6-2).

IR (Neat) : 3400 cm^-1

NMR (CDCl₃, δ) : 1.1-1.3 (3H, m) , 1.9-2.4 (3H, m), 2.7-2.9 (1H, m) , 3.0-3.2 (1H, m) ,

3.3-3.9 (6H, m), 7.1-7.5 (5H, m)

Preparation 7-3)

(2S,4S)-1-Allyloxycarbonyl-2-hydroxymethyl-4-(2-hydroxypropyDpyrrolidme was obtained in 64.7% yield in substantially the same manner as that of Preparation 6-3).

IR (Neat) : 3400, 1685 cm^-1

NMR (CDCl₃, δ) : 1.20, 1.21 (3H, each d, J=6Hz),

2.3-2.7 (3H, m), 3.07 (1H, t, J=10Hz), 3.5-4.2 (5H, m) , 4.60 (2H, d, J=6Hz), 5.1-5.4 (2H, m), 5.8-6.0 (1H, m)

Preparation 7-4)

( 2S, 4S) -1-Allyloxycarbonyl-2-carboxy-4-(2-oxopropyl)-pyrrolidine was obtained in 78.0% yield in substantially the same manner as that of Preparation 6-4).

IR (Neat) : 1710 cm^-1

NMR (CDCl₃, δ) : 2.16 (3H, s), 2.2-2.8 (4H, m),

2.9-3.1 (1H, m), 3.8-3.9 (1H, m),

4.3-4.5 (1H, m), 4.6-4.8 (2H, m),

5.1-5.5 (2H, m) , 5.8-6.0 (1H, m)

Preparation 7-5)

( 2S,4S)-1-Allyloxycarbonyl-2-(N,N-dimethylcarbamoyl)-4-(2-oxopropyl)pyrrolidine was obtained in 77.7% yield in substantially the same manner as that of Preparation 6-5).

IR (Neat) : 1700, 1650 cm^-1

NMR (CDCl₃, δ) : 2.14 (3H, s) , 2.95 (3H, s) ,

3.10 (3H, s), 3.8-4.0 (1H, s), 4.5-4.8 (3H, m) , 5.1-5.5 (2H, m) , 5.8-6.0 (1H, m)

Preparation 7-6)

(3S,4R)-4-{3-[(2S,4S)-1-Allyloxycarbonyl-2-(N,N-dirnethylcarbamoyl)pyrrolidin-4-yl]-2-oxopropyl}-3-[(1R)-1-t-butyldimethylsilyloxyethyl]-2-oxoazetidine was obtained in 82.0% yield in substantially the same manner as that of Preparation 6-6).

IR (CH₂Cl₂) : 1750, 1700, 1650 cm^-1

NMR (CDCl₃, δ) : 0.06 (6H, s) , 0.91 (9H, s),

1.19 (3H, d, J=7Hz), 2.98 (3H, s), 3.07 (3H, s),

3.8-4.0 (2H, m), 4.5-4.8 (3H, m) ,

5.1-5.4 (2H, m), 5.8-6.0 (1H, m) ,

6.05 (1H, br s) Preparation 8-1)

1-t-Butyloxycarbonyl-3-methanesulfonyloxymethyl- pyrrolidine was obtained quantitatively in substantially the same manner as that of Preparation 1-4).

IR (Neat) : 1700 cm^-1

NMR (CDCl₃, δ) : 1.46 (9H, s), 1.6-2.2 (4H, m) ,

2.5-2.8 (1H, m), 3.01 (3H, s), 3.0-3.7 (4H, m) , 4.0-4.4 (2H, m) Preparation 8-2)

To a solution of 1-t-butyloxycarbonyl-3- methanesulfonyloxymethylpyrrolidine (24 g) in dimethyl sulfoxide (200 ml) was added sodium cyanide (10 g) at ambient temperature under nitrogen. After stirring at 100ºC under nitrogen for 1 hour, the solution was taken up into a mixture of ethyl acetate and water. The organic layer was washed in turn with water and brine. The dried solvent was evaporated, and the residue was

chromatographed on silica gel eluting with a mixture of n-hexane and ethyl acetate (1:1, V/V) to give

1-t-butyloxycarbonyl-3-cyanomethylpyrrolidine (17.0 g) .

IR (Neat) : 2260, 1700 cm^-1

NMR (CDCl₃, δ) : 1.46 (9H, s) , 1.5-2.2 (2H, m) ,

2.4-2.7 (3H, m), 3.0-3.2 (1H, m) ,

3.2-3.8 (3H, m)

Preparation 8-3)

After a solution of 1-t-butyloxycarbonyl-3-cyanomethylpyrrolidine (17.0 g) in a mixture of acetic acid (70 ml) and cone, hydrochloric acid (70 ml) was stirred for 3 hours at 100°C, the solvent was evaporated in vacuo. The residue was dissolved in a mixture of water (80 ml) and tetrahydrofuran (80 ml). To the solution was dropwise added allyl chloroformate (10.3 ml) at 0ºC while adjusting pH to 8.5 with 30% aqueous sodium hydroxide solution. After stirring for 10 minutes, the solution was adjusted to pH 2.0 with 1N-hydrochloric acid. The mixture was extracted with ethyl acetate and then the organic layer was washed with brine, and dried over magnesium sulfate. The solvent was evaporated to give

allyloxycarbonyl-3-carboxymethylpyrrolidine (17.0 g).

IR (Neat) : 1720 cm^-1

NMR (CDCl₃, δ) : 1.4-1.8 (1H, m) , 2.0-2.2 (1H, m) , 2.4-2.8 (3H, m), 3.0-3.2 (1H, m) , 3.3-3.9 (3H, m), 4.61 (2H, m) , 5.1-5.4 (2H, m), 5.8-6.0 (1H, m)

Preparation 8-4)

1-Allyloxycarbonyl-3-(2-oxopropyl)pyrrolidine was obtained in 66.0% yield in substantially the same manner as that of Preparation 1-8).

IR (Neat) : 1710 cm^-1

NMR (CDCl₃, δ) : 2.16 (3H, s), 2.4-2.7 (3H, m),

2.9-3.0 (1H, m), 3.3-3.8 (3H, m) , 4.57 (2H, m), 5.1-5.4 (2H, m) , 5.8-6.0 (1H, m)

Preparation 8-5)

( 3S, 4R) -4-[3-(1-Allyloxycarbonylpyrrolidin-3-yl)-2- oxopropyl]-3-[(1R)-1-t-butyldimethylsilyloxyethyl]-2-oxoazetidine was obtained in 55.9% yield in substantially the same manner as that of Preparation 1-9) .

IR (Neat) : 3400, 1700, 1755 cm^-1

NMR (CDCl₃, δ) : 0.07 (6H, s) , 0.94 (9H, s) ,

1.16 (3H, d, J=6Hz), 2.4-3.0 (6H, m) , 4.4-4.5 (2H, m) , 5.0-5.3 (2H, m) ,

5.8-6.0 (1H, m), 6.07 (1H, br s)

Preparation 9-1)

To a solution of (2S)-1-t-butyloxycarbonyl-2-carboxy¬pyrrolidine (17.6 g) in tetrahydrofuran (200 ml) were added triethylamine (13.7 ml) and isobutyl chloroformate (11.7 ml) at -30ºC. After stirring for 30 minutes at 0ºC, the precipitate was filtered off. The obtained solution was added dropwise to the solution of sodium borohydride (7 g) in water (70 ml) at 0ºC. After stirring for 1 hour at 0°C, sodium chloride was added thereto, and the organic layer was separated. The dried organic layer was

evaporated and then the residue was chromatographed on silica gel eluting with a mixture of n-hexane and ethyl acetate (2:1 ~ 1:2, V/V) to give (2S)-1-t-butyloxycarbonyl-2-hydroxymethylpyrrolidine (14.6 g).

IR (Neat) : 3400, 1700 cm^-1

NMR (CDCl₃, δ) : 1.47 (9H, s), 1.6-2.2 (4H, m) ,

3.2-3.7 (5H, m)

Preparation 9-2)

(2S)-1-t-Butyloxycarbonyl-2-methanesulfonyloxy-methylpyrrolidine was obtained quantitatively in

substantially the same manner as that of Preparation 1-4).

IR (Neat) : 1690 cm^-1

NMR (CDCl₃,-δ) : 1.47 (9H, s), 1.7-2.2 (4H, m) ,

3.01 (3H, s), 3.35 (2H, m)

Preparation 9-3)

(2S) -1-t-Butyloxycarbonyl-2-cyanomethylpyrrolidine was obtained in 88.8% yield in substantially the same manner as that of Preparation 8-2).

IR (Neat) : 2250, 1690 cm^-1

NMR (CDCl₃, δ) : 1.47 (9H, s), 1.7-2.3 (4H, m) ,

2.5-2.9 (2H, m), 3.4 (2H, m) , 4.06 (1H, m) Preparation 9-4)

(2S) -1-Allyloxycarbonyl-2-carboxymethylpyrrolidine was obtained in 96.5% yield in substantially the same manner as that of Preparation 8-3). IR (Neat) : 1700 cm^-1

NMR (CDCl₃, δ) : 3.43 (2H, m) , 4.1-4.3 (1H, m),

4.60 (2H, d, J=5Hz), 5.1-5.3 (2H, m), 5.8-6.0 (1H, m)

Preparation 9-5)

(2S)-1-Allyloxycarbonyl-2-(2-oxopropyl)pyrrolidine was obtained in 38.7% yield in substantially the same manner as that of Preparation 1-8).

IR (Neat) : 1700 cm^-1

NMR (CDCl₃, δ) : 2.21 ( 3H, s) , 3.3-3.5 (2H, m),

4.1-4.3 (1H, m), 4.58 (2H, d, J=5Hz), 5.1-5.3 (2H, m), 5.8-6.0 (1H, m) Preparation 9-6)

(3S,4R)-4-{3-[(2S)-1-Allyloxycarbonylpyrrolidin-2-yl]-2-oxopropyl}-3-[{IR)-1-t-butyldim thylsilyloxyethyl]-2-oxoazetidine was obtained in 69.5% yield in

substantially the same manner as that of Preparation 1-9).

IR (Neat) : 3300, 1760, 1700 cm^-1

NMR (CDCl₃, δ) : 0.03 (6H, s), 0.85 (9H, s),

3.34 (2H, m), 4.4-4.6 (2H, m) , 5.1-5.3 (2H, m) , 5.8-6.0 (1H, m) Preparation 10-1)

1-t-Butyloxycarbonyl-3-methanesulfonyloxyazetidine was obtained from 1-t-butyloxycarbonyl-3-hydroxyazetidine quantitatively in substantially the same manner as that of Preparation 1-4).

IR (CH₂Cl₂) : 1710 cm^-1

NMR (CDCl₃, δ) : 1.44 (9H, s), 3.06 (3H, s),

4.0-4.4 (4H, m), 5.1-5.3 (1H, m)

Preparation 10-2)

1-t-Butyloxycarbonyl-3-cyanoazetidine was obtained in 80.0% yield in substantially the same manner as that of Preparation 8-2).

IR (Neat) : 2250, 1700 cm^-1

NMR (CDCl₃, δ) : 1.44 (9H, s), 3.3-3.4 (1H, m),

4.0-4.3 (4H, m)

Preparation 10-3)

1-t-Butyloxycarbonyl-3-carboxyazetidine was obtained quantitatively in substantially the same manner as that of Preparation 8-3).

IR (Neat) : 1700 cm^-1

NMR (CDCl₃, δ) : 1.44 (9H, s), 4.0-4.2 (4H, m)

Preparation 10-4)

l-t-Butyloxycarbonyl-3-(hydroxymethyl)azetidine was obtained in 98.8% yield in substantially the same manner as those of Preparation 1-2) and 1-3).

IR (Neat) : 3400, 1680 cm^-1

NMR (CDCl₃, δ) : 1.42 (9H, s), 3.7-4.2 (4H, m)

Preparation 10-5)

1-t-Butyloxycarbonyl-3-(methanesulfonyloxymethyl)-azetidine was obtained quantitatively in substantially the same manner as that of Preparation 1-4).

IR (Neat) : 1690 cm^-1

NMR (CDCl₃, δ) : 1.44 ( 9H, s) , 3.03 (3H, s),

3.9-4.2 (4H, m)

Preparation 10-6)

1-t-Butyloxycarbonyl-3-(cyanomethyl)azetidine was obtained in 56.6% yield in substantially the same manner as that of Preparation 8-2).

IR (Neat) : 2250, 1700 cm^-1

NMR (CDCl₃, δ) : 1.44 (9H, s) , 2.64 (2H, d, J=7Hz), 2.7-3.0 (1H, m) , 3.6-3.7 (2H, m) , 4.0-4.2 (2H, m) Preparation 10-7)

1-Allyloxycarbonyl-3-(carboxymethyl)azetidine was obtained quantitatively in substantially the same manner as that of Preparation 8-3).

IR (Neat) : 1700 cm^-1

NMR (CDCl₃, δ) : 2.68 (2H, d, J=7Hz ) , 2.8-3.1 (1H, m), 3.6-3.8 (2H, m) , 4.1-4.3 (2H, m) , 4.5-4.6 (2H, m), 5.1-5.4 (2H, m) , 5.8-6.0 (1H, m) Preparation 10-8)

1-Allyloxycarbonyl-3-(2-oxopropyl)azetidine was obtained in 53.6% yield in substantially the same manner as that of Preparation 1-8).

IR (Neat) : 1700 cm^-1

NMR (CDCl₃, δ) : 2.15 (3H, s), 2.7-3.1 (3H, m) ,

3.3-3.6 (2H, m) , 4.0-4.2 (2H, m) ,

4.5-4.6 (2H, m), 5.1-5.4 (2H, m) ,

5.8-6.0 (1H, m) Preparation 10-9)

( 3S, 4R) -4-[ 3-(1-Allyloxycarbonylazetidin-3-yl)-2-oxopropyl]-3-[(1R)-1-t-butyldimethylsilyloxyethyl]-2-oxoazetidine was obtained in 26.1% yield in substantially the same manner as that of Preparation 1-9).

IR (Neat) : 1760, 1700 cm^-1

NMR (CDCl₃, δ) : 0.06 (6H, s) , 0.91 (9H, s),

1.21 (3H, d, J=6Hz), 2.6-3.0 (6H, m), 3.5-3.7 (2H, m), 4.0-4.3 (3H, m) ,

4.5-4.6 (2H, m), 5.1-5.3 (2H, m),

5.8-6.0 (1H, m), 6.04 (1H, br s)

Preparation 11

To a suspension of cyanoacetamide (5.0 g) in ethanol (3.5 ml) was introduced hydrogen chloride (2.2 g) at 0°C. The resultant solid was allowed to stand at 0ºC for 48 hours to give ethyl carbamoylacetimidate hydrochloride (9.87 g).

IR (Nujol) : 1680, 1650 cm^-1 Preparation 12-1)

A mixture of cyanoacεtic acid (15.0 g) , dicyclohexyl carbodiimide (35.4 g) , dimethylamine hydrochloride (14.4 g) and triethylamine (24.6 ml) in dichloromethane (300 ml) was stirred at ambient temperature for 12 hours. The resultant precipitate was filtered off, and the filtrate was concentrated in vacuo. The residue was

chromatographed on silica gel eluting with a mixture of n-hexane and ethyl acetate (6:4 ~ 0:10, V/V) to give dimethylcarbamoylacetonitrile (16.09 g) .

IR (Nujol) : 1660 cm^-1

NMR (CDCl₃, δ) : 3.00 (3H, s), 3.08 (3H, s),

3.50 (2H, s)

Preparation 12-2)

To a solution of dimethylcarbamoylacetonitrile (15.0 g) in a mixture of ethanol (7.85 ml) and chloroform (10 ml) was introduced hydrogen chloride (4.88 g) at 0ºC. The resultant mixture was allowed to stand at 0ºC for 24 hours. The precipitates were collected by filtration, washed with diethyl ether and dried in vacuo to give ethyl dimethylcarbamoylacetimidate hydrochloride (27.10 g).

NMR (DMSO-d₆, δ) : 1.36 (3H, t, J=7Hz ) , 2.86 (3H, s), 3.00 (3H, s), 4.01 (2H, s), 4.50 (2H, q, J=7Hz), 12.0 (1H, br s) Preparation 13-1)

A mixture of 4-bromobutyronitrile (35 ml) in benzene (175 ml) and pyridine (35 ml) was heated at 70ºC for 5 hours. The resulting crystals were filtered, washed with diethyl ether and dried under reduced pressure to give 4-(1-pyridinio)butyronitrile bromide (5.74 g).

NMR (DMSO-d₆, δ) : 2.00-3.00 (4H, m) , 4.80 (2H, t, J=7Hz), 8.00-8.35 (2H, m) , 8.35-8.80 (1H, m) , 8.95-9.32 (2H, m)

Preparation 13-2)

Ethyl 4-(1-pyridinio)butanoimidate

bromide·hydrochloride was obtained quantitatively

according to a similar manner to that of Preparation 11.

NMR (DMSO-d₆, δ) : 1.36 (3H, t, J=7Hz), 2.00-2.85

(4H, m), 4.43 (2H, q, J=7Hz), 4.60-4.80 (2H, m) , 8.00-9.25 (5H, m)

Preparation 14)

3-Pyridylacetonitrile (5.0 g) was dissolved in a mixture of ethanol (2.50 ml) and chloroform (30 ml) and hydrogen chloride gas (3.12 g) was introduced therein at 0°C. After leaving the reaction mixture at 0ºC for 12 hours, the resulting crystals were filtered, washed with diethyl ether and dried under reduced pressure to give ethyl 3-pyridylacetimidate·hydrochloride (12.85 g).

NMR (DMSO-d₆, δ) : 1.30 (3H, t, J=7Hz) , 4.26-4.70 (4H, m), 7.80-8.13 (1H, m) , 8.35-8.62 (1H, m) , 8.70-9.02 (2H, m) , 10.5 (2H, br s)

Preparation 15-1)

Methyl iodate (13.2 ml) was added to a acetone solution (20 ml) containing 3-pyridilacetonitrile (5.0 g) and the mixture was allowed to stand at ambient

temperature for 15 hours. The resulting crystals were filtered, washed with acetone and diethyl ether, and dried under reduced pressure to give 3-cyanomethyl-l-methyl- pyridinium iodide (10.57 g).

NMR (DMSO-d₆, δ) : 4.37 (3H, s), 4.39 (2H, s),

8.10-8.25 (1H, m) , 8.50-8.65 (1H, m) , 8.95-9.10

(2H, m)

Preparation 15-2)

Ethyl 1-methyl-3-pyridinioaσetimidate

iodide·hydrochloride was obtained quantitatively according to a similar manner to that of Preparation 11.

NMR (DMSO-d₆, δ) : 1.00-1.50 (3H, m) , 3.30-4.80 (7H, m), 6.85-9.20 (6H, m) Preparation 16-1)

To a stirred solution of N-benzyloxycarbonylglycine (100 g) and triethylamine (86.6 ml) in tetrahydrofuran (1000 ml) was added isobutyl chloroformate (73.0 ml) at -20 ~ -15°º for 15 minutes. To the mixture was added a solution of 3-amino-1-propanol (54.9 ml) in

tetrahydrofuran (50 ml) at -55 ~ -50°C for 30 minutes.

The reaction mixture was poured into water (2 ℓ) and ethyl acetate (3 ℓ), and the organic layer was separated. The organic layer was washed in turn with lN-aqueous

hydrochloric acid (1 ℓ), lN-aqueous sodium hydroxide (1 ℓ) , water ( 1 ℓ) , and brine (1 ℓ). Removal of the solvents gave N-benzyloxycarbonylglycine 3-hydroxypropylamide (90.5 g).

NMR (DMSO-d₆, δ) : 1.54 (2H, tt, J=6.0Hz, 6.4Hz), 3.11 (2H, dt, J=6.0Hz, 6.4Hz), 3.40 (2H, dt,

J=5.3Hz, 6.0Hz), 3.57 (2H, d, J=6.3Hz), 4.43 (1H, t, J=5.3Hz), 5.03 (2H, s), 7.35 (5H, s), 7.42 (1H, t, J=6.0Hz), 7.81 (1H, t, J=6.0Hz) Preparation 16-2) To a stirred suspension of sodium borohydride (33.6 g) in tetrahydrofuran (1.5 ℓ) was added boron trifluoride diethyl etherate (109 ml) at 5-10ºC for 20 minutes, and the mixture was stirred for 30 minutes at the same temperature. To the mixture was added

N-benzyloxycarbonylglycine 3-hydroxypropylamide (94.5 g) under 15ºC for 15 minutes and the mixture was stirred for 1 day at ambient temperature. To the mixture was added concentrated hydrochloric acid (140 ml) at 5-10°C and the mixture was stirred at ambient temperature for 2 hours. To the stirred mixture was added water (500 ml) and carbobenzoxy chloride (40.5 ml) at 5-10°C while keeping pH to 9.5-10.5 with IN aqueous sodium hydroxide, and the resulting mixture was stirred for 30 minutes at 5-10ºC. The reaction mixture was extracted with ethyl acetate (1ℓ, × 2) and the organic layer was washed in turn with

lN-aqueous hydrochloric acid (500 ml), saturated aqueous sodium hydrogen carbonate (500 ml) and brine (500 ml). Removal of the solvents gave a residue which was

chromatographed on silica gel (2000 ml) eluting with a mixture of n-hexane and ethyl acetate (5:5 - 1:9, V/V) to give a mixture of 4,7-bis(benzyloxycarbonyl)-4,7-diazaheptan-1-ol and 4,7-bis(benzyloxycarbonyl)-4,7-diazaoctan-1-ol (89.4 g) (3:2).

IR (Film) : 3750-3100, 1690 cm^-1

NMR (CDCl₃, δ) : 1.44-1.94 (2H, m) , 2.90 (1.2H, s), 3.12-3.70 (8H, m) , 5.03 (2H, s), 5.06 (2H, s), 7.26 (10H, s) Preparation 16-3)

To a stirred solution of 4,7-bis(benzyloxycarbonyl)-4,7-diazaheptan-1-ol and 4,7-bis(benzyloxycarbonyl)-4,7-diazaoctan-1-ol (total 89.4 g) in acetone (1000 ml) was added Jones' reagent (2.67 mmol/ml) (140 ml) at 30-40°C, and the mixture was stirred for 30 minutes. To the resulting mixture was added isopropyl alcohol (5 ml), and the mixture was chromatographed on Florisil (Trademark, made by Floridin Co.) (100-200 mesh) (300 ml) eluting with ethyl acetate to give a mixture of

3-[N-(benzyloxycarbonyl)-N-{2-(benzyloxycarbonylamino)-ethyl}amino]propionic acid and 3-[N-(benzyloxycarbonyl)-N-{2-(N-benzyloxycarbonyl-N-methylamino)ethyl}amino]- propionic acid (94.5 g) (3:2).

IR (Film) : 3700-2700, 1690 cm^-1

NMR (CDCl₃, δ) : 2.57 (2H, t, J=6Hz), 2.92 (1.2H, s), 3.10-3.66 (6H, m) , 5.02 (2H, s), 5.05 (2H, s), 5.60-6.55 (1H, br s), 7.25 (10H, s)

Preparation 16-4)

A suspension of 3-[N-(benzyloxycarbonyl)-N-{2- (benzyloxycarbonylamino)ethyl}amino]propionic acid and 3-[N-(benzyloxycarbonyl)-N-{2-(N-benzyloxycarbonyl-N-methylamino)ethyl}amino]propionic acid (total 94.5 g) and 10% Pd-C (50% wet) (37 g) in methanol (1 ℓ) was stirred at ambient temperature for 2.5 hours under hydrogen

atmosphere. The catalyst was removed by filtration and to the filtrate was added water (1 ℓ). To the resulting solution was added allyloxycarbonyl chloride (50 ml) at 5-10°C while keeping pH to 9.5-10.5 with IN aqueous sodium hydroxide, and the resulting mixture was stirred for 30 minutes at 5-10ºC. The resulting solution was washed with ethyl acetate (500 ml). To the aqueous layer was added ethyl acetate (1 ℓ) and pH of the mixture was adjusted to 2 with 6N-aqueous hydrochloric acid. The organic layer was separated and removal of the solvents gave a mixture of 3-[N-(allyloxycarbonyl)-N-{2-(allyloxycarbonylamino)-ethyl}amino]propionic acid and 3-[N-(allyloxycarbonyl)-N-{2-(N-allyloxycarbonyl-N-methylamino)ethyl}amino]propionic acid (total 58.8 g) (3:2).

IR (Film) : 3700-2750, 1690 cm^-1 NMR ( CDCl₃ , δ ) : 2.55-2.76 ( 2H, m) , 2.96 ( 1. 2H, s) , 3 . 24-3 . 66 ( 6H, m) , 4. 45-4. 64 ( 4H, m) , 5 . 10-5 . 40 ( 4H, m) , 5.78-6. 05 ( 2H, m) Preparation 16-5)

A mixture of 4-[N-(allyloxycarbonyl)-N-{2- (allyloxycarbonylamino)ethyl}amino]butan-2-one and 4-{N- (allyloxycarbonyl)-N-{2-(N-allyloxycarbonyl-N-methylamino)ethyl}amino]butan-2-one (total 41.0 g) (3:2) was obtained from 3-[N-(allyloxycarbonyl)-N-{2- (allyloxycarbonylamino)ethyl}amino]propionic acid and 3-[N-(allyloxycarbonyl)-N-{2-(N-allyloxycarbonyl-N-methylamino)ethyl}amino]propionic acid in substantially the same manner as that of Preparation 1-8).

IR (CH₂Cl₂) : 1700 cm^-1

NMR (CDCl₃, δ) : 2.14 (3H, s) , 2.62 (2H, t, J=8Hz), 2.91 (1.2H, s), 3.25-3.60 (6H, m) , 4.45-4.70 (4H, m), 5.05-5.40 (4H, m), 5.62-6.13 (2H, m) Preparation 16-6)

To a stirred solution of 4-[N-(allyloxycarbonyl)-N- {2- (allyloxycarbonylamino)ethyl}amino]butan-2-one and 4-[N-(allyloxycarbonyl)-N-{2-(N-allyloxycarbonyl-N-methyl¬amino)ethyl}amino]butan-2-one (total 39.0 g) and

1-ethylpiperidine (46.8 ml) in dichloromethane (300 ml) was added trimethylsilyl trifluoromethanesulfonate (63.3 ml) at -20ºC, and the resulting mixture was stirred at 0°C for 2 hours. (Solution A) To a solution of

(3R,4R)-4-acetoxy-3-[(1R)-1-t-butyldimethylsilyloxyethyl]-2-oxoazetidine (75.3 g) and 1-ethylpiperidine (36.0 ml) in dichloromethane (300 ml) was added trimethylsilyl

trifluoromethanesulfonate (50.6 ml) at -20ºC2 To this mixture were added zinc bromide (118.0 g) and Solution A at 0ºC, and the resulting mixture was stirred for 1.5 hours at ambient temperature. The reaction mixture was taken up into a mixture of ethyl acetate (2.1ℓ) and water (1.4ℓ). After adjusting pH to 0.7 with 6N-aqueous hydrochloric acid, the mixture was stirred for 1 hour. After adjusting pH of the mixture to 7 with 4N-aqueous sodium hydroxide, the organic layer was separated, washed with brine and dried over magnesium sulfate. Removal of the solvents gave a residue, which was chromatographed on silica gel (2.5 ℓ) eluting with a mixture of n-hexane and ethyl acetate (4:6-2:8, V/V) to give a mixture of (3S,4R)- [4-[N-(allyloxycarbonyl)-N-{2-(allyloxycarbonylamino)-ethyl}amino]-2-oxobutyl]-3-[(1R)-1-t-butyldimethylsilyloxyethyl]-2-oxoazetidine and (3S,4R)-4-[4-[N- (allyloxycarbonyl) -N-{2-(N-allyloxycarbonyl-N-methyl- amino)ethyl}amino]-2-oxobutyl]-3-[(1R)-1-t-butyldimethyl- silyloxyethyl]-2-oxoazetidine (total 35.6 g) (3:2).

IR (CH₂Cl₂) : 1760, 1700 cm^-1

NMR (CDCl₃, δ) : 0.07 (6H, s) , 0.97 (9H, s),

2.54-3.05 (5H, m) , 2.92 (1.2H, s), 3.20-3.70 (6H, m), 3.80-4.24 (2H, m) , 4.42-4.71 (4H, m), 4.96-5.44 (4H, m) , 5.63-6.35 (3H, m)

Preparation 16-7)

A mixture of allyl (5R,6S)-3-[2-[N-(allyloxycarbonyl-N-{2-(allyloxycarbonylamino)ethyl}amino]ethyl]-6-[(1R)-1-t-butyldimethylsilyloxyethyl]-7-oxo-l-azabicyclo[3.2.0]-hept-2-ene-2-carboxylate and allyl (5R,6S)-3-[2-[N-(allyloxycarbonyl)-N-{2-(N-allyloxycarbonyl-N-methylamino)ethyl}amino]ethyl]-6-[(1R)-1-t-butyldimethylsilyloxyethyl]-7-oxo-1-azabicyclo[3.2.0]hept-2-ene-2-carboxylate (total 35.0 g) was obtained from (3S,4R)-4-[4-[N-(allyloxycarbonyl)-N-{2-(allyloxycarbonylamino)-ethyl}amino]-2-oxobutyl]-3-[(1R)-1-t-butyldimethylsilyloxyethyl]-2-oxoazetidine and ( 3S,4R)-4-[4-[N-(allyloxycarbonyl)-N-{2-(N-allyloxycarbonyl-N-methylamino)ethyl}- amino]-2-oxobutyl]-3-[(1R)-1-t-butyldimethylsilyloxyethyl]-2-oxo-azetidine (total 35.0 g) in substantially the same manner as that of Preparation 1-10). The obtained mixture was chromatographed on silica gel (2.5 ℓ) eluting with a mixture of acetone and dichloromethane (0:100 - 11:89, V/V) to give allyl ( 5R,6S) -3-[2- [N-( allyloxycarbonyl) -N-{2-(allyloxycarbonylamino)ethyl}amino]-ethyl]-6-[(1R)-1-t-butyldimethylsilyloxyethyl]-7-oxo-1-azabicyclo[3.2.0]hept-2-ene-2-carboxylate (17.0 g).

IR (Film) : 1780, 1700 cm^-1

NMR (CDCl₃, δ) : 0.07 (6H, s) , 0.88 (9H, s), 1.24 (3H, d, J=6.2Hz), 2.63-3.19 (5H, m) , 3.25-3.60 (6H, m), 4.01-4.24 (2H, m) , 4.45-4.83 (6H, m), 5.18-5.48 (6H, m) , 5.79-6.04 (3H, m)

Elution was continued to give allyl (5R,6S)-3-[2-[N- (allyloxycarbonyl)-N-{2-(N-allyloxycarbonyl-N-methylamino)ethyl}amino]ethyl]-6-(1R)-1-t-butyldimethylsilyloxyethyl]-7-oxo-1-azabicyclo[3.2.0]hept-2-ene-2-carboxylate (9.4 g) .

IR (Film)- : 1780, 1700 cm^-1

NMR (CDCl₃, δ) : 0.07 (6H, s), 0.88 (9H, s),

1.24 (3H, d, J=6.3Hz), 2.63-3.19 (5H, m),

2.92 (3H, s), 3.28-3.57 (6H, m) , 4.00-4.27 (2H, m), 4.49-4.84 (6H, m) , 5.14-5.46 (6H, m),

5.81-6.05 (3H, m)

Preparation 16-8)

Allyl (5R,6S)-3-[2-[N-(allyloxycarbonyl)-N-{2-(allyloxycarbonylamino)ethyl}amino]ethyl]-6-[(1R)-1-hydroxyethyl]-7-oxo-l-azabicyclo[3.2.0]hept-2-ene-2-carboxylate (8.13 g) was obtained in substantially the same manner as that of Preparation 1-11).

IR (Film) : 3650-2750, 1770, 1700 cm^-1

NMR (CDCl₃, δ) : 1.34 (3H, d, J=6.2Hz) , 2.67-3.l63 ( 11H, m) , 4. 04-4.32 ( 2H, m) , 4. 48-5 . 05 ( 6H, m) , 5 . 14-5. 52 ( 6H, m) , 5 . 80-6 . 05 ( 3H, m)

Preparation 16-9)

Allyl (5R,6S)-3-[2-[N-(allyloxycarbonyl)-N-{2-(N- allyloxycarbonyl-N-methylamino)ethyl}amino]ethyl]-6-[(1R)- 1-hydroxyethyl]-7-oxo-1-azabicyclo[3.2.0]hept-2-ene-2- carboxylate (3.68 g) was obtained in substantially the same manner as that of Preparation 1-11).

IR (Film) : 3700-2800, 1770, 1690 cm^-1

NMR (CDCl₃, δ) : 1.33 (3H, d, J=6.3Hz), 2.60-3.67 (11H, m), 2.92 (3H, s), 4.04-4.32 (2H, m),

4.42-4.89 (6H, m), 5.13-5.52 (6H, m), 6.79-6.08 (3H, m)

Preparation 17-1)

N-(2-Hydroxyethyl)-3-(benzyloxycarbonylamino)-propanamide (124.7 g) was obtained in substantially the same manner as that of Preparation 16-1).

NMR (DMSO-d₆, δ) : 2.26 (2H, t, J=7.3Hz), 3.01-3.42

(6H, m), 4.64 (1H, t, J=5.6Hz), 5.00 (2H, s), 7.23 (1H, t, J=5.6Hz), 7.34 (5H, s), 7.86 (1H, t, J=5.0HZ) Preparation 17-2)

A mixture of 3,7-bis(benzyloxycarbonyl)-3,7-diazaheptan-1-ol and 3,7-bis(benzyloxycarbonyl)-3,7- diazaoctan-1-ol (total 145 g) (1:1) was obtained in substantially the same manner as that of Preparation

16-2).

IR (Film) : 3400, 1680 cm^-1

NMR (CDCl₃, δ) : 1.70-2.00 (2H, m) , 2.85 (1.5H, s), 2.90-3.82 (8H, m) , 5.12 (4H, s), 7.34 (10H, s) Preparation 17-3) A mixture of 2-[N-(benzyloxycarbonyl)-N-{3-(benzyloxycarbonylamino)propyl}amino]acetic acid and

2-[N-(benzyloxycarbonyl)-N-{3-(N-benzyloxycarbonyl-N- methylamino)propyl}amino]acetic acid (total 146 g) (1:1) was obtained in substantially the same manner as that of Preparation 16-3).

IR (Film) : 3700-2550, 1690 cm^-1

NMR (CDCl₃, δ) : 1.49-1.98 (2H, m) , 2.86 (1.5H, s), 3.03-3.52 (4H, m) , 3.99 (2H, s) , 5.10 (4H, s), 7.27 (10H, s)

Preparation 17-4)

A mixture of 2-[N-(allyloxycarbonyl)-N-{3-(allyloxycarbonylamino)propyl}amino]acetic acid and

2-[N- (allyloxycarbonyl)-N-{3-(N-allyloxycarbonyl-N-methylamino)propyl}amino]acetic acid (total 100 g) (1:1) was obtained in substantially the same manner as that of Preparation 16-4).

IR (Film) : 3750-2450, 1690 cm^-1

NMR (CDCl₃, δ) : 1.62-1.95 (2H, m) , 2.92 (1.5H, s),

3.08-3.53 (4H, m) , 4.02 (2H, d, J=6.7Hz),

4.56 (4H, d, J=4.lHz), 5.02-5.40 (4H, m) ,

5.40-5.65 (1H, br s) Preparation 17-5)

A mixture of 3-[N-(allyloxycarbonyl) -N-{3-(allyloxy¬carbonylamino)propyl}amino]propan-2-one and 3-[N-(allyl¬oxycarbonyl) -N-{3-(N-allyloxycarbonyl-N-methylamino)propyl}amino]propan-2-one (total 61.4 g) (1:1) was obtained in substantially the same manner as that of Preparation 16-5).

IR (CH₂Cl₂) : 1700, 1690 cm^-1

NMR (CDCl₃, δ) : 1.55-1.90 (2H, m) , 2.14 (3H, s) ,

2.91 (1.5H, s), 3.12-3.43 (4H, m) , 4.3-4.7 (4H, m), 5.05-5.43 (4H, m) , 5.66-6.10 (2H, m) Preparation 17-6)

A mixture of ( 3S,4R)-4-[3-[N-(allyloxycarbonyl)-N- {3-(allyloxycarbonylamino)propyl}amino]-2-oxopropyl]-3- [(1R) -1-t-butyldimethylsilyloxyethyl]-2-oxoazetidine and ( 3S, 4R) -4-[3-[N-(allyloxycarbonyl)-N-{3-(N-allyloxy- carbonyl-N-methylamino)propyl}amino]-2-oxopropyl]-3-[(1R) 1-t-butyldimethylsilyloxyethyl]-2-oxoazetidine (total 92.52 g) (1:1) was obtained in substantially the same manner as that of Preparation 16-6).

NMR (CDCl₃, δ) : 0.07 (6H, s), 0.87 (9H, s),

1.21 (3H, d, J=6Hz), 1.57-1.80 (2H, m) ,

2.52-2.93 (3H, m) , 2.90 (1.5H, s), 3.17-3.29 (2H, m), 3.29-3.43 (2H, m) , 3.88-4.26 (4H, m) , 4.52-4.66 (4H, m) , 5.13-5.39 (4H, m) , 5.73-6.09 (2H, m)

Preparation 17-7)

A mixture of allyl (5R,6S)-3-[[N- (allyloxycarbonyl)-N-{3-(allyloxycarbonylamino)propyl}- amino]methyl]-6-[(1R)-1-t-butyldimethylsilyloxyethyl]-7-oxo-1-azabicyclo[3.2.0]hept-2-ene-2-carboxylate and allyl ( 5R, 6S) -3-[ [N-(allyloxycarbonyl-N-{3-(N-allyloxy- carbony- 1-N-methylamino)propyl}amino]methyl]-6-[(1R)-1-t-butyldimethylsilyloxyethyl]-7-oxo-l-azabicyclo[3.2.0]hept- 2-ene-2-carboxylate (total 59.13 g) (1:1) was obtained in substantially the same manner as that of Preparation

16-7).

IR (Film) : 1780, 1700 cm^-1

NMR (CDCl₃, δ) : 0.07 (6H, s), 0.88 (9H, s),

1.23 (3H, d, J=6Hz), 1.63-1.88 (2H, m) ,

2.73-3.40 (7H, m) , 2.90 (Ϊ.5H, s), 4.02-4.84 (10H, m), 5.17-5.52 (6H, m) , 5.78-6.07 (3H, m)

Preparation 17-8)

A mixture of allyl (5R,6S)-3-[[N-(allyloxy carbonyl)-N-{3-(allyloxycarbonylamino)propyl}amino]methyl]-6-[(1R)-1-hydroxyethyl]-7-oxo-1-azabicyclo[3.2.0]hept-2-ene-2-carboxylate and allyl (5R,6S)-3-[[N-(allyloxy-carbonyl)-N-{3-(N-allyloxycarbonyl-N-methylamino)propyl}amino]methyl] -6-[ ( IR) -1-hydroxyethyl] -7-oxo-l-azabicyclo[3.2.0]hept-2-ene-2-carboxylate (total 7.96 g) (1:1) was obtained in substantially the same manner as that of Preparation 16-8).

NMR (CDCl₃, δ) : 1.33 (3H, d, J=6.3Hz), 1.60-1.94 (2H, m), 2.79-3.44 (7H, m) , 2.90 (1.5H, s),

4.07-5.08 (10H, m), 5.13-5.57 (7H, m) , 5.79-6.08 (3H, m)

Preparation 18-1)

To a solution of 2- (2-aminoethylamino)ethanol (100 g) in a mixture of tetrahydrofuran (500 ml) and water (500 ml) was added dropwise at 10ºC a solution of allyl

chloroformate (230 ml) in tetrahydrofuran (100 ml) while adjusting. pH to around 10 with 4N-aqueous sodium

hydroxide. After stirring at 10°C for 30 minutes, to the mixture was added sodium chloride and ethyl acetate ( 1 ℓ) , and the organic layer was separated. The organic solution was washed successively with lN-hydrochloric acid (300 ml), water (300 ml) and brine (300 ml), and dried over magnesium sulfate. Removal of the solvents gave

3 ,6-bis( allyloxycarbonyl)-3,6-diazahexan-1-ol ( 266.2 g).

IR (Film) : 3650-2700, 1680 cm^-1

NMR (CDCl₃, δ) : 2.61 (2H, br s), 3.18-3.56 (6H, m), 3.67-3.86 (2H, m), 4.45-4.66 (4H, m) , 5.07-5.38 (4H, m), 5.76-6.04 (2H, m)

Preparation 18-2)

To a solution of 3 ,6-bis(allyloxycarbonyl)-3,6-diazahexan-1-ol (100.0 g) in dichloromethane (800 ml) were added successively t-butyldimethylsilyl chloride (58.1 g), triethylamine (56.3 ml), and 1,8-diazabicyclo[5.4.0]undec-7-ene (5.5 ml). After stirring at ambient temperature for 18 hours, the solution was washed successively with

1N-aqueous sodium hydroxide (500 ml), saturated aqueous sodium hydrogen carbonate (300 ml) and brine (300 ml), and dried over magnesium sulfate. Removal of the solvents gave a residue, which was chromatographed on silica gel (2 ℓ) eluting with a mixture of n-hexane and ethyl acetate (95:5 - 70:30, V/V) to give 3,6-bis(allyloxycarbonyl)-1- (t-butyldimethylsilyloxy)-3,6-diazahexane ( 129.3 g).

IR (Film) : 3500-3100, 1680 cm^-1

NMR (CDCl₃, δ) : 0.05 (6H, s), 0.89 (9H, s),

3.26-3.54 (6H, m) , 3.65-3.83 (2H, m), 4.59-4.63 (4H, m) , 5.14-5.48 (3H, m),

5.82-6.05 (2H, m)

Preparation 18-3)

To a suspension of sodium hydride (60% dispersion in mineral oil) (18.16 g) in a mixture of tetrahydrofuran (800 ml) and N,N-dimethyIformamide (400 ml) were added successively at 5-10°C propargyl bromide (25.1 ml), and a solution of 3 ,6-bis(allyloxycarbonyl)-1-(t-butyldimethylsilyloxy)-3,6-diazahexane (117.0 g) in tetrahydrofuran (100 ml) and N,N-dimethylformamide (50 ml). After stirring at the same temperature for 30 minutes, to the mixture was added 3% aqueous ammonium chloride, and the organic layer was separated. The organic solution was washed successively with water (500 ml) and brine (500 ml), and dried over magnesium sulfate. Removal of the solvents gave a residue, which was

chromatographed on silica gel (2 ℓ) eluting with a mixture of n-hexane and ethyl acetate (95:5 - 80:20, V/V) to give 4,7-bis(allyloxycarbonyl)-9-(t-butyldimethylsilyloxy)-4,7-diazanon-1-yne (107.1 g). IR (CH₂Cl₂) : 3350-3150, 1690 cm^-1

NMR (CDCl₃, δ) : 0.04 (6H, s), 0.84 (9H, s),

2.20 (1H, s), 3.26-3.40 (2H, m) , 3.46-3.56 (4H, m), 3.60-3.76 (2H, m) , 4.00-4.17 (2H, m),

4.49-4.61 (4H, m), 5.11-5.34 (4H, m) , 5.77-6.03

(2H, m),

Preparation 18-4)

A suspension of 4,7-bis(allyloxycarbonyl)-9-(t-butyl- dimethylsilyloxy)-4,7-diazanon-1-yne (49.40 g),

mercury(II) sulfate (5.19 g) , Florisil (trademark)

(100-200 mesh) (17.3 g) and sulfuric acid (1.75 ml) in tetrahydrofuran (500 ml) and water (100 ml) was stirred at 50°C for 5 hours. The mixture was chromatographed on Florisil (500 ml) eluting with ethyl acetate. Removal of the solvents gave a residue which was chromatographed on silica gel (1.3 ℓ) eluting with a mixture of n-hexane and ethyl acetate (70:30 - 0:100, V/V) to give 3-[N-(allyloxycarbonyl)-N-[2-{N-allyloxycarbonyl-N-(2-hydroxyethyl)-amino}ethyl]amino]propan-2-one (26.81 g).

IR (CH₂Cl₂) : 3700-3250, 1690 cm^-1

NMR (CDCl₃, δ) : 2.13 (3H, s), 3.35-3.58 (6H, m), 3.64-3.84 (2H, m), 3.96-4.18 (2H, m), 4.43-4.67 (4H, m), 5.12-5.42 (4H, m), 5.75-6.06 (2H, m)

Preparation 18-5)

3-[N-(Allyloxycarbonyl)-N-[2-{N-allyloxycarbonyl- N-(2-t-butyldimethylsilyloxyethyl)amino}ethyl]amino]¬proρan-2-one (12.84 g) was obtained in substantially the same manner as that of Preparation 18-2).

IR (CH₂Cl₂) : 1690 cm^-1

NMR (CDCl₃, δ) : 0.04 (6H, s), 0.88 (9H, s),

2.14 (3H, s), 3.28-3.57 (6H, m) , 3.64-3.83 (2H, m), 3.96-4.14 (2H, m) , 4.49-4.67 (4H, m) , 5.12-5. 42 ( 4H, m) , 5. 67-6.06 ( 2H, m)

Preparation 18-6)

( 3S,4R)-4-[3-{N-(Allyloxycarbonyl)-N-[2-{N-allyloxy- carbonyl-N-(2-t-butyldimethylsilyloxyethyl)amino}ethyl]- amino]-2-oxopropyl]-3-[(1R)-1-t-butyldimethylsilyloxyethyl]-2-oxoazetidine (18.50 g) was obtained in

substantially the same manner as that of Preparation 16-6).

IR (CH₂Cl₂) : 1760, 1690 cm^-1

NMR (CDCl₃, δ) : 0.03 (6H, s), 0.07 (6H, s),

0.87 (9H, s), 0.88 (9H, s), 1.17 (3H, d,

J=6.2Hz), 2.48-2.88 (3H, m) , 3.24-3.52 (6H, m), 3.61-3.77 (2H, m) , 3.89-4.24 (4H, m) , 4.48-4.61

(4H, m), 5.10-5.36 (4H, m) , 5.71-5.99 (2H, m)

Preparation 18-7)

Allyl (5R,6S)-3-[N-(allyloxycarbonyl)-N-[2-{N-allyloxycarbonyl-N-(2-t-butyldimethylsilyloxyethyl)amino}-ethyl]amino]methyl-6-[(1R)-1-t-butyldimethylsilyloxyethyl]-7-oxo-1-azabicyclo[3.2.0]hept-2-ene-2-carboxylate (14.40 g) was obtained in substantially the same manner as that of Preparation 16-7).

IR (CH₂Cl₂) : 1780, 1700 cm^-1

NMR (CDCl₃, δ) : 0.04 (6H, s), 0.07 (6H, s),

0.88 (18H, s), 1.22 (3H, d, J=6.lHz), 2.68-3.12 (3H, m), 3.04-3.82 (8H, m) , 4.04-4.33 (2H, m) , 4.52-4.97 (8H, m) , 5.15-5.52 (6H, m) , 5.80-6.08 (3H, m)

Preparation 18-8)

Allyl ( 5R, 6S)-3-[N-(allyloxycarbonyl)-N-[2-{N-allyl¬oxycarbonyl-N-(2-hydroxyethyl)amino}-ethyl]amino]methyl-6-[(1R)-l-hydroxyethyl]-7-oxo-1- azabicyclo[3.2.0]hept-2-ene-2-carboxylate (4.74 g) was obtained in substantially the same manner as that of Preparation 16-8).

IR (CH₂Cl₂) : 1780, 1690 cm^-1

NMR (CDCl₃, δ) : 1.33 (3H, d, J=6.1Hz),

1.57-2.15 (3H, m), 2.68-3.88 (10H, m), 4.08-4.32 (2H, m), 4.32-4.91 (8H, m), 5.14-5.54 (6H, m), 5.79-6.11 (3H, m) Preparation 19-1)

Ethyl (3R)-3-allyloxycarbonylamino-4-azidobutanoate (52.0 g) was added to a stirred solution of methyl iodide (13.5 ml) in N,N-dimethylacetamide (350 ml) at -20ºC.

Then sodium hydride (10.5 g) was added portionwise at the same temperature. After stirring for 2 hours, the

reaction was quenched with water. The reaction mixture was poured into isopropyl ether (IPE, 1 ℓ) and water (1.4 ℓ). The organic layer was separated and aqueous layer was extracted with IPE (350 ml × 2). The combined organic layer was washed in turn with water (500 ml × 4), and brine (500 ml × 1), and dried over magnesium sulfate.

Evaporation of the solvent gave crude ethyl (3R)-3-[N- (allyloxycarbonyl)-N-methylamino]-4-azidobutanoate (50.7 g). The product was used in the next reaction without further purification.

NMR (CDCl₃, δ) : 1.30 (3H, t, J=6.0Hz), 2.5-2.8 (2H, m), 2.93 (3H, s), 3.20-3.75 (2H, m) , 4.16 (2H, q, J=6Hz), 4.0-4.35 (1H, m) , 4.40-4.73 (2H, m), 5.0-5.46 (2H, m), 5.60-6.20 (1H, m)

Preparation 19-2)

Ethyl (3R)-3-[N-(allyloxycarbonyl)-N-methylamino]-4-azidobutanoate (60.5 g) was dissolved in brine-saturated ethyl acetate (600 ml), and triphenylphosphine (51.0 g) was added thereto. Stirring was continued for 8 hours, and water (250 ml) was added. After adjusting pH to 1.5 with hydrochloric acid, the organic layer was separated. The aqueous layer was washed with ethyl acetate (100 ml × 2). Tetrahydrofuran (200 ml) was added to the aqueous phase and the aqueous phase was cooled to 0ºC. A solution of allyl chloroformate (12.4 ml) was added thereto while adjusting pH to around 9 with a solution of sodium

hydroxide. The reaction mixture was diluted with ethyl acetate (750 ml), washed in turn with water, hydrochloric acid (1N), saturated aqueous sodium hydrogen carbonate and brine, and dried over magnesium sulfate. Evaporation of the solvent gave ethyl (3R)-3-[N-(allyloxycarbonyl)-N-methylamino]-4-(allyloxycarbonylamino)butanoate (50.7 g).

NMR (CDCl₃, δ) : 1.27 (3H, t, J=7.2Hz), 2.45-2.74 (2H, m), 2.85 (3H, s), 3.19-3.30 (2H, m) , 4.10

(2H, q, J=7.2Hz), 4.40-4.70 (5H, m) , 5.18-5.40 (4H, m), 5.80-6.05 (2H, m)

Preparation 19-3)

A solution of sodium hydroxide (2N, 161 ml) was added to a stirred solution of ethyl (3R)-3-[N-(allyloxycarbonyl)-N-methylamino]-4-(allyloxycarbonylamino)butanoate (50.7 g) in ethanol (500 ml). After stirring at room temperature for 2 hours, the solvent was evaporated. The residue was diluted with ethyl acetate (150 ml) and water (300 ml), and the organic layer was separated. The aqueous layer was diluted with ethyl acetate (400 ml) and adjusted to pH 1.5 with hydrochloric acid. The organic layer was separated, washed with brine and dried over magnesium sulfate. Evaporation of the solvent gave

(3R)-3-[N-(allyloxycarbonyl)-N-methylamino]-4-(allyloxycarbonylaminoJbutanoic acid (46.6 g).

NMR (CDCl₃, δ) : 2.47-2.80 (2H, m) , 2.86 (3H, s), 3.20-3.60 (2H, m) , 4.40-4.70 (4H, m), 5.15-5.40 (4H, m) , 5.75-6.05 (2H, m), 7.4-8. 4 ( 1H, br s )

Preparation 19-4)

Meldrum's acid (25.6 g), dicyclohexylcarbodiimide (36.7 g) and 4-dimethylaminopyridine (21.7 g) were added successively to a stirred suspension of

(3R)-3-[N-(allyloxycarbonyl)-N-methylamino]-4- (allyloxycarbonylamino)butanoic acid (46.6 g) in

dichloromethane (450 ml) at 5ºC. After stirring at room temperature for 10 hours, the mixture was washed in turn with hydrochloric acid (IN, 100 ml), cold water (100 ml) and brine (100 ml), and the solvent was evaporated. The residue was diluted with 50% acetic acid (500 ml) and stirred for 10 hours at 70ºC. Evaporation of the solvent gave a crude oil which was diluted with ethyl acetate (500 ml) and washed in turn with saturated aqueous sodium hydrogen carbonate and brine, and dried over magnesium sulfate. After evaporation of the solvent, the residue was chromatographed on silica gel (10 g,

eluent:n-hexane/ethyl acetate = 9/1-6/4) to afford

(4R)-4-[N-(allyloxycarbonyl)-N-methylamino]-5- (allyloxycarbonylamino)pentan-2-one (4.37 g).

NMR (CDCl₃, δ) : 2.08 (3H, s), 2.38-2.85 (2H, m), 3.74 (3H, s), 3.0-3.38 (2H, m), 3.85-4.30 (1H, m), 4.30-4.60 (4H, m) , 4.70-5.30 (4H, m),

5.54-6.04 (2H, m)

Preparation 19-5)

To a stirred mixture of ( 4R) -4-[N-(allyloxycarbonyl)-N-methylamino]-5-(allyloxycarbonylamino)pentan-2-one (4.37 g), sodium iodide (0.22 g) and triethylamine (9.75 ml) in acetonitrile (100 ml) was added chlorotrimethylsilane (8.35 ml) at room temperature. After stirring for 1 hour at room temperature, the solvent was evaporated. n-Hexane (50 ℓ) was added to the residue and the precipitate was filtrated off. After the filtrate was concentrated in vacuo, the residue was diluted with ethyl acetate (50 ml) and poured into a mixture of (3R,4R)-4-acetoxy-3-[(1R)-1- t-butyldimethylsilyloxyethyl]azetidin-2-one (5.03 g) and dried zinc bromide (3.30 g) in ethyl acetate (50 ml) at room temperature. Stirring was continued for 8 hours, then the mixture was poured into water (50 ml), whose pH was adjusted to around 1.5 with hydrochloric acid (6N). After stirring for 2 hours, the organic layer was

separated, washed in turn with 10% aqueous sodium hydrogen carbonate (50 ml) and brine (50 ml × 1), and dried over magnesium sulfate. After evaporation of the solvent, the residue was chromatographed on silica gel (100 g,

eluent:n-hexane/ethyl acetate = 9/1-0/10) to afford

(3S,4R)-4-[(4R)-4-{N-(allyloxycarbonyl)-N-methylamino}-5- (allyloxycarbonylamino)-2-oxopentyl]-3-[(1R)-1-t-butyldimethylsilyloxyethyl]-2-oxoazetidine (1.93 g).

NMR (CDCl₃, δ) : 0.6 (6H, s), 0.85 (9H, s), 1.17

(3H, d, J=6Hz), 2.60-2.86 (7H, m) , 3.13-3.43 (2H, m), 3.76-4.30 (2H, m) , 4.33-4.65 (5H, m),

4.96-5.40 (4H, m), 5.56-6.10 (2H, m)

Preparation 19-6)

Allyl oxalyl chloride (0.68 ml) was added to a stirred solution of (3S,4R)-4-[(4R)-4-{N-(allyloxycarbonyl)-N-methylamino}-5-(allyloxycarbonylamino)-2-oxopentyl]-3-[(1R)-1-t-butyldimethylsilyloxyethyl]-2-oxoazetidine (1.81 g) and triethylamine (0.96 ml) in dichloromethane (18 ml) at -20ºC, and the resulting mixture was stirred for 30 minutes at the same

temperature. The reaction mixture was poured into a mixture of ethyl acetate (60 ml) and water (40 ml). The organic layer was separated, washed in turn with water, aqueous sodium hydrogen carbonate and brine, and dried over magnesium sulfate. After evaporation of the solvent. the residue was dissolved in triethyl phosphite (1.77 ml), and stirred at 90ºC under nitrogen atmosphere. After stirring for 2 hours, hydroquinone (568 mg) and toluene (20 ml) were added, and the resulting mixture was heated to reflux for additional 1.5 hours. After being cooled to room temperature, the reaction mixture was diluted with ethyl acetate (60 ml) and washed in turn with 30% aqueous sodium carbonate (30 ml × 5) and brine (30 ml × 1), and dried over magnesium sulfate. After evaporation of the solvent, the residue was chromatographed on silica gel (20 g, eluent:n-hexane/ethyl acetate = 9/1-7/3) to afford allyl (5R,6S)-3-[(2R)-2-{N-(allyloxycarbonyl)-N-methylamino}-3-(allyloxycarbonylamino)propyl]-6-[(1R)-1-t-butyl¬dimethyl-silyloxyethyl]-7-oxo-1-azabicyclo[3.2.0]hept-2-ene-2-carboxylate (2.2 g).

NMR (CDCl₃, δ) : 0.06 (6H, s), 0.90 (9H, s),

2.45-3.45 (5H, m) , 2.75 (3H, s), 3.71-4.23 (3H, m), 4.23-4.76 (6H, m) , 5.10-5.43 (6H, m),

5.73-6.10 (3H, m)

Preparation 19-7)

To a stirred solution of allyl (5R,6S)-3-[(2R)-2-{N- (allyloxycarbonyl)-N-methylamino}-3-(allyloxycarbonylamino)propyl]-6-[(1R)-1-t-butyldimethylsilyloxyethyl]-7-oxo-1-azabicyclo[3.2.0]hept-2-ene-2-carboxylate (2.2 g) and acetic acid (0.8 ml) in tetrahydrofuran (20 ml) was added 70% aqueous tetrabutylammonium fluoride (3.5 g) .

After stirring at room temperature for 15 hours, the reaction mixture was poured into ethyl acetate (80 ml) and water (60 ml). The organic layer was separated and washed in turn with water (20 ml × 4), aqueous sodium hydrogen carbonate (30 ml × 1) and brine, and dried over magnesium sulfate. After evaporation of the solvent, the residue was chromatographed on silica gel (40 ml) eluting with a mixture of n-hexane and ethyl acetate to afford allyl ( 5R, 6S)-3-[(2R)-2-{N-(allyloxycarbonyl)-N-methylamino}-3- (allyloxycarbonylamino)propyl]-6-[(1R)-1-hydroxyethyl]-7-oxo-1-azabicyclo[3.2.0]hept-2-ene-2-carboxylate (600 mg).

NMR (CDCl₃, δ) : 1.32 (2H, d, J=6Hz), 2.30-3.46 (5H, m), 2.78 (3H, s), 3.80-4.96 (11H, m) , 5.0-5.53 (6H, m), 5.60-6.18 (3H, m)

Preparation 20-1)

Ethyl (3S)-3-allyloxycarbonylamino-4-azidobutanoate (44.5 g) was added to a stirred solution of methyl iodide (19.2 ml) in N,N-dimethylformamide (300 ml) at 0°C. Then sodium hydride (11.9 g) was added portionwise at the same temperature. After stirring for 2 hours, the reaction was quenched with water. The reaction mixture was poured into ethyl acetate (1.2 ℓ) , and water (0.9 ℓ). The organic layer was separated and aqueous layer was extracted with ethyl acetate (300 ml × 2). The combined organic layer was washed in turn with water (300 ml x 4) and brine (300 ml × 1), and dried over magnesium sulfate. Evaporation of the solvent gave crude ethyl (3S)-[N-(allyloxycarbonyl)-N-methylamino]-4-azidobutanoate (48.6 g). The product was used in the next reaction without further purification.

Preparation 20-2)

Ethyl ( 3S)-3-[N-(allyloxycarbonyl)-N-methylamino]-4- (allyloxycarbonylamino)butanoate was obtained in

substantially the same manner as that of Preparation

19-2).

NMR (CDCl₃, δ) : 1.27 (3H, t, J=7.2Hz), 2.45-2.74 ( 2H, m) , 2. 85 ( 3H, s ) , 3 .19-3 . 30 ( 2H, m) , 4.10 ( 2H, q, J=7. 2Hz ) , 4. 40-4.70 ( 5H, m) , 5. 18-5 . 40 ( 4H, m) , 5 .80-6.05 ( 2H, m) Preparation 20-3)

(3S)-3-[N-(Allyloxycarbonyl)-N-methylamino]-4- (allyloxycarbonylamino)butanoic acid was obtained in substantially the same manner as that of Preparation 19-3).

NMR (CDCl₃, δ) : 2.47-2.80 (2H, m) , 2.86 (3H, s),

3.20-3.60 (2H, m), 4.40-4.70 (4H, m), 5.15-5.40 (4H, m), 5.75-6.05 (2H, m)

Preparation 20-4)

( 4S) -4-[N-(Allyloxycarbonyl)-N-methylamino]-5- (allyloxycarbonylamino)pentan-2-one was obtained in substantially the same manner as that of Preparation 19-4).

NMR (CDCl₃, δ) : 2.08 (3H, s) , 2.38-2.85 (2H, m), 3.74 (3H, s), 3.0-3.38 (2H, m), 3.85-4.30 (1H, m), 4.30-4.60 (4H, m) , 4.70-5.30 (4H, m) , 5.54-6.04 (2H, m)

Preparation 20-5)

(3S,4R)-4-[(4S)-4-{N-(Allyloxycarbonyl)-N-methylamino}-5-(allyloxycarbonylamino)-2-oxopentyl]-3-[(1R)-1-t-butyldimethylsilyloxyethyl]-2-oxoazetidine was obtained in substantially the same manner as that of Preparation 19-5).

NMR (CDCl₃, δ) : 0.06 (6H, s) , 0.88 (9H, s), 1.18

(3H, d, J=6.0Hz), 2.60-2.86 (7H, m) , 3.13-3.43 (2H, m), 3.76-4.30 (2H, m) , 4.33-4.65 (5H, m) , 4.96-5.40 (4H, m) , 5.56-6.10 (2H, m) Preparation 20-6) Allyl ( 5R,6S) -3-[(1S)-1-{N-(allyloxycarbonyl)-N- methylamino}-2-(allyloxycarbonylamino)ethyl]-6-[(1R)-1- t-butyldimethylsilyloxyethyl]-7-oxo-l-azabicyclo[3.2.0]- hept-2-ene-2-carboxylate was obtained in substantially the same manner as that of Preparation 19-6).

NMR (CDCl₃, δ) : 0.07 (6H, s), 0.88 (9H, s),

2.45-3.46 (5H, m) , 2.75 (3H, s), 3.80-4.96 (11H, m), 5.0-5.53 (6H, m), 5.70-6.03 (3H, m) Preparation 20-7)

Allyl ( 5R,6S)-3-[(1S)-1-{N-(allyloxycarbonyl)-N-methylamino}-2-(allyloxycarbonylamino)ethyl]-6-[(1R)-1-hydroxyethyl]-7-oxo-l-azabicyclo[3.2.0]hept-2-ene-2-carboxylate was obtained in substantially the same manner as that of Preparation 19-7).

NMR (CDCl₃, δ) : 1.31 (2H, d, J=6.5Hz), 2.30-3.46 (5H, m), 2.75 (3H, s), 3.80-5.00 (1H, m),

5.0-5.60 (6H, m), 5.60-6.18 (3H, m) Preparation 21

To a stirred solution of methylthioacetonitrile (7.27 g) in dichloromethane (140 ml) was added by portions m-chloroperbenzoic acid (17.26 g) at 0ºC. After stirring at 0ºC for 30 minutes, the resulting precipitate was filtered off. Evaporation of the solvent gave a residue which was chromatographed on silica gel (300 ml) eluting with a mixture of dichloromethane and acetone (10:0 - 4:6, V/V) to give methylsulfinylacetonitrile (8.29 g).

IR (Nujol) : 2230 cm^-1

NMR (CDCl₃, δ) : 2.85 (3H, s), 3.67 (1H, d, J=16Hz),

3.87 (1H, d, J=16Hz)

Preparation 22

To a stirred solution of chloroacetonitrile (3 ml) and 1-methyl-5-mercaptotetrazole (5.50 g) in dichloromethane (30 ml) was added triethylamine (13 ml) at 0ºC. After stirring at 0ºC for 1 hour, the reaction mixture was diluted with ethyl acetate (100 ml) and the resulting precipitate was filtrate off. The filtrate was washed in turn with brine, hydrochloric acid ( 6N) , aqueous potassium carbonate (30%) and brine, and dried over magnesium sulfate. Evaporation of the solvent gave

5-cyanomethylthio-1-methyltetrazole (5.66 g).

NMR (CDCl₃, δ) : 4.01 (3H, s), 4.17 (2H, s)

Preparation 23-1)

(2-Hydroxyethylthio)acetonitrile (4.47 g) was

obtained in substantially the same manner as that of

Preparation 27.

IR (Nujol) : 3350, 2240 cm^-1

NMR (CDCl₃, δ) : 2.42 (1H, s), 2.92 (2H, t, J=6Hz),

3.42 (2H, s), 3.90 (2H, t, J=6Hz)

Preparation 23-2)

To a stirred solution of (2-hydroxyethylthio)acetonitrile (4.46 g) in dichloromethane (30 ml) was added dropwise trichloroacetyl isocyanate (4.5 ml) at 0°C.

After stirring at ambient temperature for 30 minutes, the reaction mixture was evaporated in vacuo. The residue was taken up into methanol (100 ml) and to the solution was added Amberlyst 15 (Trademark, made by Rohm & Haas Co.) (H-form, dry, 40 g) . The mixture was heated to reflux for 16 hours, and the resin was filtered off. The filtrate was evaporated, and the residue was chromatographed on silica gel (100 ml) eluting with a mixture of n-hexane and ethyl acetate (9:1 - 7:3, V/V) to give ( 2-carbamoyloxyethylthio) acetonitrile (5.32 g) .

IR (Nujol) : 3420, 3320, 3250, 3200, 2250, 1690 cm^-1 NMR (CDCl₃, δ) : 3.00 (2H, t, J=6Hz), 3.40 (2H, s),

4.32 (2H, t, J=6Hz) Preparation 24-1)

A suspension of 2-bromo-N,N-dimethylacetamide (10.0 g) and potassium thioacetate (8.26 g) in acetonitrile (100 ml) was stirred at ambient temperature for 1 hour. The resulting presipitate was filtered off, and the filtrate was evaporated. The residue was taken up into ethyl acetate (100 ml), washed with brine and dried over

magnesium sulfate. Evaporation of the solvent gave

2-acetylthio-N,N-dimethyl acetamide (7.74 g).

NMR (CDCl₃, δ) : 2.39 (3H, s), 2.98 (3H, s),

3.10 (3H, s), 3.82 (2H, s)

Preparation 24-2)

To a solution of 2-acetylthio-N,N-dimethylacetamide (7.74 g) in methanol (50 ml) was added dropwise a solution of sodium methoxide in methanol (4.8N, 10 ml) at 0ºC.

After stirring at -15ºC for 15 minutes, to the mixture was added chloroacetonitrile (3.04 ml) at 0ºC. After stirring at 0°C for 30 minutes, solvents were evaporated and the residue was taken up into ethyl acetate (100 ml), washed with brine (10 ml), dried over magnesium sulfate.

Evaporation of the solvent gave a residue which was chromatographed on silica gel (150 ml) eluting with a mixture of n-hexane and ethyl acetate (9:1 - 3:7, V/V) to give 2-(cyanomethylthio)-N,N-dimethylacetamide (7.06 g).

IR (Nujol) : 2250, 1630 cm^-1

NMR (CDCl₃, δ) : 3.00 (3H, s), 3.10 (3H, s),

3.30-3.70 (4H, m) Preparation 25-1)

To a solution of 1-methanesulfonyloxy-2-methoxyethane (17.74 g) in N,N-dimethylformamide (140 ml) was added potassium thioacetate (15.77 g), and the resulting mixture was heated to 70ºC for 2 hours. The reaction mixture was taken up into a mixture of water (300 ml) and ethyl acetate (700 ml). The organic layer was separated, washed in turn with water (150 ml × 4) and brine, and dried over magnesium sulfate. Evaporation of the solvent gave a residue which was chromatographed on silica gel (300 ml) eluting with a mixture of n-hexane and ethyl acetate

(10:0 - 94:6, V/V) to give 1-acetylthio-2-methoxyethane (3.62 g).

NMR (CDCl₃, δ) : 2.35 (3H, s), 3.09 (2H, t, J=6Hz),

3.35 (3H, s), 3.52 (2H, t, J=6Hz)

Preparation 25-2)

To a solution of 1-acetylthio-2-methoxyethane (5.77 g) in methanol (33 ml) was added dropwise a solution of sodium methoxide in methanol (4.8N, 8.96 ml) at 0ºC.

After stirring for 30 minutes at 0ºC, to the solution was added chloroacetonitrile (2.74 ml) at 0ºC. After stirring at 0ºC for another 30 minutes, the reaction mixture was diluted with ethyl acetate (200 ml), washed with brine, dried over magnesium sulfate. Evaporation of the solvent gave a residue which was chromatographed on silica gel (300 ml) eluting with a mixture of n-hexane and ethyl acetate (10:0 - 9:1, V/V) to give (2-methoxyethylthio)- acetonitrile (2.52 g).

IR (Nujol) : 2250 cm^-1

NMR (CDCl₃, δ) : 2.90 (2H, t, J=6Hz), 3.36 (3H, s),

3.41 (2H, s), 3.69 (2H, t, J=6Hz)

Preparation 26

t-Butylthioacetonitrile (10.60 g) was obtained in substantially the same manner as that of Preparation 27.

IR (Nujol) : 2240 cm^-1

NMR (CDCl₃, δ) : 0.92 (9H, s) , 2.78 (2H, s)

Preparation 27 To a stirred solution of chloroacetonitrile (2.8 ml) and 2-propene-1-thiol (3.5 ml) in dichloromethane (40 ml) was added triethylamine (6.7 ml) at 0ºC. After stirring at 0ºC for 2 hours, the reaction mixture was diluted with ethyl acetate (200 ml), washed with brine, dried over magnesium sulfate and evaporated. The residue was chromatographed on silica gel (100 ml) eluting with a mixture of n-hexane and ethyl acetate (95:5 - 2:1, V/V) to give allylthioacetonitrile (2.11 g).

NMR (CDCl₃, δ) : 2.20 (2H, s), 2.32 (2H, d, J=7Hz),

5.00-5.40 (2H, m), 5.45-6.00 (1H, m)

Preparation 28

To a stirred solution of 2-mercapto-1-methylimidazole (10.0 g) in dichloromethane (50 ml) were added

triethylamine (28.3 ml) and chloroacetonitrile (5.54 ml) at 0°C. After stirring at 0ºC for 6 hours, the reaction mixture was diluted with ethyl acetate (300 ml) and the resulting precipitate was filtered off. The filtrate was concentrated in vacuo to give crude

1-methyl-2-(cyanomethylthio)imidazole (14.0 g) which was used for the next experiment without further purification.

NMR (DMSO-d₆, δ) : 3.66 (3H, s), 4.11 (2H, s),

7.04 (1H, d, J=1.2Hz), 7.35 (1H, d, J=1.2Hz)

Preparation 29

To a stirred solution of aminoacetonitrile

hydrochloride (10.0g) in dichloromethane (200 ml) were added successively pyridine (20.1 ml) and methanesulfonyl chloride (8.79 ml). Evaporation of the solvent gave a residue which was chromatographed on silica gel (1 £) eluting with a mixture of ethyl acetate and methanol

(10:0 - 9:1, V/V) to give (methanesulfonylamino)-acetonitrile (13.32 g).

NMR (DMSO-d₆, δ) : 3.01 (3H, s), 4.18 (2H, s), 7.96 (1H, br s)

Preparation 30

To a stirred solution of aminoacetonitrile

hydrochloride (25.0 g) in a mixture of water (200 ml) and tetrahydrofuran (300 ml) was added acetyl chloride (39 ml) at 0ºC while adjusting pH to around 9.5 with aqueous sodium hydroxide (4N). After evaporation of the solvent, the residue was extracted with ethyl acetate (1 ℓ × 2). The combined extract was evaporated in vacuo to give

(acetylamino)acetonitrile (25.78 g).

NMR (DMSO-d₆, δ) : 1.88 (3H, s) , 4.10 (2H, d,

J=5.2Hz), 8.57 (1H, br s)

Preparation 31

To a solution of aminoacetonitrile hydrochloride (5.0 g) in a mixture of water (40 ml) and tetrahydrofuran (10 ml) was added potassium cyanate (6.6 g) by portions at 50ºC and the resultant mixture was heated at 50ºC for 3 hours. To the mixture was added dropwise hydrochloric acid (1N, 20 ml) at 50ºC and heated at 50ºC for 30

minutes. After complete evaporation of the solvent, the residue was extracted twice with methanol (30 ml).

Evaporation of the solvent gave (cyanomethyl)urea (4.29 g).

IR (Nujol) : 3400, 2260, 1660, 1610 cm^"1

NMR (DMSO-d₆, δ) : 4.00 (2H, d, J=5Hz),

5.88 (2H, br s)

Preparation 32

To a stirred solution of aminoacetonitrile

hydrochloride (10 g) in a mixture of water (100 ml) and tetrahydrofuran (150 ml) was added methyl chloroformate (10 ml) at 0ºC while adjusting pH to around 8.5 with aqueous sodium hydroxide (4N). After stirring for another 30 minutes at 0ºC, the solvents were evaporated, and the residue was extracted with ethyl acetate (500 ml).

Extracts were evaporated to give (methoxycarbonylamino)- acetonitrile (12.0 g).

NMR (DMSO-d₆, δ) : 3.61 (3H, s), 4.08 (2H, d,

J=6Hz), 7.90 (1H, br s)

Preparation 33

To a stirred solution of 3-hydroxypropanenitrile (8.6 ml) in dichloromethane (50 ml) was added dropwise

trichloroacetyl isocyanate (15.0 ml) at around 10ºC.

After stirring at ambient temperature for 30 minutes, the reaction mixture was evaporated. The residue was taken up into methanol (150 ml) and to the solution was added

Amberlyst 15 (H-form, dry, 6.0 g). The mixture was heated to reflux for 8 hours and the resin was filtered off.

After evaporation of the solvent, the residue was

crystallized from dichloromethane to give

3-carbamoyloxypropanenitrile (11.04 g).

NMR (DMSO-d₆, δ) : 2.82 (2H, t, J=6Hz), 4.08 (2H, t,

J=6Hz), 6.68 (2H, br s)

Preparation 34

To a solution of ethyl isocyanate (3.75 ml) in dichloromethane (30 ml) was added hydroxyacetonitrile (3.0 ml) at 0ºC. The resulting mixture was allowed to stand at ambient temperature for 40 hours, and heated to 40ºC for 2 hours. The solvent and the excess hydroxyacetonitrile were removed by distillation under reduced pressure to give (N-ethylcarbamoyloxy)acetonitrile (4.44 g).

NMR (CDCl₃, δ) : 1.16 (3H, t, J=7Hz), 3.00-3.44 (2H, m), 4.70 (2H, s), 5.01 (1H, br s)

Preparation 35-1)

To a solution of methylsulfinylacetonitrile (1.0 g) in methanol (9.7 ml) was added a solution of sodium methoxide in methanol (4.8N, 0.2ml). After standing at ambient temperature for 12 hours, to the solution was added acetic acid (0.055 ml) and the mixture was

evaporated in vacuo. The residue was taken up into dichloromethane (10 ml) and the resulting precipitate was filtered off. Evaporation of the solvent gave crude methyl methylsulfinylacetimidate (containing about 25% of starting material) (1.02 g) , which was used for the next step without further purification.

IR (Neat) : 3250, 1650 cm^-1

NMR (CDCl₃, δ) : 2.63 (3H, s), 3.56 (2H, s),

3.74 (3H, s)

The following compounds were obtained in

substantially the same manner as that of Preparation

35-1).

Preparation 35-2)

Ethyl ( 1-methyltetrazol-5-ylthio)acetimidate

hydrochloride

NMR (DMSO-d₆, δ) : 1.28 (3H, t, J=7Hz), 4.00 (3H, s), 4.46 (2H, q, J=7Hz) , 4.47 (2H, s),

Preparation 35-3)

Ethyl methylthioacetimidate hydrochloride

NMR (DMSO-d₆, δ) : 1.41 (3H, t, J=7Hz), 2.26 (3H, s), 3.73 (2H, s), 4.52 (2H, q, J=7Hz)

Preparation 35-4)

Ethyl (2-carbamoyloxyethylthio)acetimidate

hydrochloride

NMR (DMSO-d₆, δ) : 1.39 (3H, t, J=7Hz), 2.89 (2H, t, J=6Hz), 3.78 (2H, s), 4.06 (2H, t, J=6Hz),

4.48 (2H, q, J=7Hz) Preparation 35-5)

Ethyl (N,N-dimethylcarbamoylmethylthio)acetimidate hydrochloride

NMR (DMSO-d₆, δ) : 1.38 (3H, t, J=7Hz), 2.84 (3H, s), 3.01 (3H, s), 3.20-3.85 (4H, m) , 4.45 (2H, q, J=7Hz)

Preparation 35-6)

Ethyl (2-methoxyethylthio)acetimidate hydrochloride NMR (DMSO-d₆, δ) : 1.35 (3H, t, J=7Hz), 2.82 (2H, t,

J=6Hz), 3.23 (3H, s), 3.49 (2H, t, J=6Hz),

3.75 (2H, s), 4.48 (2H, q, J=7Hz)

Preparation 35-7)

Ethyl (t-butylthio)acetimidate hydrochloride

NMR (DMSO-d₆, δ) : 1.00-1.50 (12H, m) , 3.81 (2H, s),

4.48 (2H, d, J=7Hz)

Preparation 35-8)

Ethyl (allylthio)acetimidate hydrochloride

NMR (DMSO-d₆, δ) : 1.39 (3H, t, J=7Hz), 3.35 (2H, d, J=7Hz), 3.68 (2H, s), 4.50 (2H, q, J=7Hz),

4.80-5.40 (2H, m) , 5.45-6.05 (1H, m) Preparation 35-9)

Ethyl ( 1-methylimidazol-2-ylthio)acetimidate

hydrochloride

NMR (DMSO-d₆, δ) : 1.14 (3H, t, J=7Hz), 3.84 (3H, s), 4.06 (2H, q, J=7Hz), 4.29 (2H, s), 7.74 (1H, d, J=1.2Hz), 7.85 (1H, d, J=1.2Hz)

Preparation 35-10)

Ethyl (methanesulfonylamino) acetimidate hydrochloride NMR (DMSO-d₆, δ) : 1.36 (3H, t, J=7Hz), 3.04 (3H, s) , 4.20 ( 2H, d, J=6Hz ) , 4.50 ( 2H, q, J=7Hz ) , 8. 14 ( 1H, t, J=6Hz )

Preparation 35-11)

Ethyl (acetylamino) acetimidate hydrochloride

NMR (DMSO-d₆, δ) : 1.37 (3H, t, J=7Hz), 1.91 (3H, s), 4.13 (2H, d, J=5.5Hz), 4.50 (2H, q, J=7Hz), 8.78 (1H, t, J=5.5Hz) Preparation 35-12)

Ethyl ureidoacetimidate hydrochloride

IR (Nujol) : 3350, 1680 cm^-1

Preparation 35-13)

Ethyl (methoxycarbonylamino)acetimidate hydrochloride NMR (DMSO-d₆, δ) : 1.34 (3H, t, J=7Hz), 3.59 (3H, s), 4.09 (2H, d, J=6Hz), 4.50 (2H, q, J=7Hz), 7.92 (1H, t, J=6Hz) Preparation 35-14)

Ethyl 3-carbamoyloxypropionimidate hydrochloride NMR (DMSO-d₆, δ) : 1.36 (3H, t, J=7Hz), 2.92 (2H, t, J=6Hz), 4.20 (2H, t, J=6Hz), 4.42 (2H, q, J=7Hz) Preparation 35-15)

Ethyl 3-methoxypropionimidate hydrochloride

NMR (DMSO-d₆, δ) : 1.35 (3H, t, J=7Hz), 2.89 (2H, t, J=6Hz), 3.25 (3H, s), 3.50 (2H, t, J=6Hz),

4.44 (2H, q, J=7Hz)

Preparation 35-16)

Ethyl hydroxyacetimidate hydrochloride

NMR (DMSO-d₆, δ) : 1.39 (3H, t, J=7Hz), 4.36 (2H, s), 4.54 (2H, q, J=7Hz) Preparation 35-17)

Ethyl 3-hydroxypropionimidate hydrochloride

NMR (DMSO-d₆, δ) : 1.40 (3H, t, J=7Hz), 2.80 (2H, t, J=6Hz), 3.73 (2H, t, J=6Hz), 4.48 (2H, q, J=7Hz)

Preparation 35-18)

Ethyl carbamoyloxyacetimidate hydrochloride

NMR (DMSO-d₆, δ) : 1.38 (3H, t, J=7Hz), 4.55 (2H, q,

J=7Hz), 4.86 (2H, s)

Preparation 35-19)

Ethyl (N-ethylcarbamoyloxy) acetimidate hydrochloride NMR (DMSO-d₆, 6) : 0.85-1.55 (6H, m), 2.80-3.20 (2H, m), 3.90-4.95 (4H, m)

Preparation 35-20)

Ethyl methoxyacetimidate hydrochloride

IR (Nujol) : 1660 cm^-1 Preparation 36-1)

To a solution of (2S,4R)-1-t-butyloxycarbonyl-4-t- butyldimethylsilyloxy-2-methoxymethylpyrrolidine (57 g) in tetrahydrofuran (280 ml) were added acetic acid (19 ml) and tetrabutylammonium fluoride (330 ml, IM solution in tetrahydrofuran) at ambient temperature. After stirring for 6 hours, the solution was poured into a mixture of ethyl acetate and water. The organic layer was washed in turn with 1N-hydrochloric acid, saturated sodium

bicarbonate, and brine. The solvent was dried over

magnesium sulfate and evaporated to give

( 2S,4R)-1-t-butyloxycarbonyl-4-hydroxy-2-methoxymethylpyrrolidine (34.7 g).

IR (Neat) : 3400, 1675 cm^-1

NMR (CDCl₃, δ) : 1.37 (9H, s), 1.8-2.1 (2H, m) , 3.25 (3H, s), 3.3-3.6 (4H, m) , 3.8-4.1 (1H, m) , 4. 2-4 . 4 ( 1H, m)

Preparation 36-2)

To a solution of oxalyl chloride (20 ml) in

dichloromethane was added dropwise a solution of dimethyl sulfoxide (20 ml) in dichloromethane at -78ºC. After stirring at -78ºC for 10 minutes, to the mixture was added dropwise a solution of (2S,4R)-1-t-butyloxycarbonyl-4- hydroxy-2-methoxymethylpyrrolidine (33 g) in

dichloromethane. After stirring at -78ºC for 10 minutes, to the mixture was added dropwise triethylamine (79 ml) and the resulting mixture was allowed to stand at 0ºC.

The solution was washed with water and dried over

magnesium sulfate. After evaporation of the solvent, the residue was chromatographed on silica gel eluting with a mixture of n-hexane and ethyl acetate (2:1 ~ 1:1, V/V) to give (2S) -1-t-butyloxycarbonyl-2-methoxymethyl-4-oxopyrrolidine (20.05 g).

IR (Neat) : 1760, 1690 cm^-1

NMR (CDCl₃, δ) : 1.49 (9H, s) , 2.4-2.8 (2H, s) , 3.31

(3H, s), 3.39 (1H, dd, J=2Hz, 9Hz), 3.5-4.0 (3H, m), 4.3-4.5 (1H, m)

Preparation 36-3)

To a suspension of sodium hydride (60%, 4.9 g) in dimethoxyethane was added dropwise triethyl

phosphonoacetate (24.1 ml). After stirring for 10

minutes, a solution of (2S)-1-t-butyloxycarbonyl-2-methoxymethyl-4-oxopyrrolidine (19 g) in dimethoxyethane was added under nitrogen at 0ºC. After stirring for 1 hour at the same temperature, the solvent was evaporated and the residue was dissolved in ethyl acetate. The organic layer was washed in turn with lN-hydrochloric acid, saturated sodium bicarbonate and brine. Evaporation of the dried extract gave ethyl (2S)-1-t-butyloxycarbonyl- 2-methoxymethylpyrrolidine-Δ ^4,α-acetate (20.3 g).

IR (Nujol) : 1690 cm^-1

NMR (CDCl₃, δ) : 1.2-1.4 (3H, m), 1.48 (9H, s),

4.0-4.4 (3H, m), 5.6-5.9 (1H, m)

Preparation 36-4)

Ethyl (2S)-1-t-butyloxycarbonyl-2-methoxymethyl-pyrrolidine-Δ ^4,α-acetate (20 g) was dissolved in ethanol and hydrogenated at room temperature under atmospheric pressure over 10% Pd/C (5 g, 50% wet). After stirring for

2 hours, the catalyst was filtered off and the filtrate was concentrated to give (2S)-1-t-butyloxycarbonyl-2-methoxymethyl-4-(ethoxycarbonylmethyl)pyrrolidine (19 g) .

IR (Neat) : 1730, 1690 cm^-1

NMR (CDCl₃, δ) : 1.25 (3H, t, J-=7Hz), 1.46 (9H, s),

3.34 (3H, s), 4.13 (2H, q, J=7Hz)

Preparation 36-5)

To a solution of (2S)-1-t-butyloxycarbonyl-2-methoxymethyl-4-(ethoxycarbonylmethyl)pyrrolidine (18 g) in ethanol (200 ml) was added 6N-hydrochloric acid (40 ml). After stirring for two days, the solvent was

evaporated, and the residue was dissolved in a mixture of tetrahydrofuran and water. To the mixture was added dropwise allyl chloroformate (8.6 ml) while adjusting pH to 10 with 30% aqueous sodium hydroxide at 0°C. After stirring for 2 hours at room temperature, the solvent was evaporated, and then the residue was partitioned between ether and water. The aqueous layer was acidified with cone, hydrochloric acid and extracted with ethyl acetate. Evaporation of the dried extract gave

(2S)-1-allyloxycarbonyl-2-methoxymethyl-4-(carboxymethyl)-pyrrolidine (19.08 g).

IR (Neat) : 1680-1710 cm^-1 NMR ( CDCl₃ , δ ) : 3.34 ( 3H, s) , 3 .3-3.8 ( 3H, m) ,

3 . 9-4.2 ( 2H, m) , 4. 59 ( 2H, d, J=5Hz ) , 5 . 1-5 . 4 ( 2H, m) , 5 . 8-6. 0 ( 1H, m) Preparation 36-6)

To a solution of (2S)-1-allyloxycarbonyl-2- methoxymethyl-4-(carboxymethyl)pyrrolidine (19.08 g) in dichloromethane (200 ml) were added

dicyclohexylcarbodiimide (16.8 g). Meldrum's acid (11.7 g) and 4-dimethylaminopyridine (9.9 g) at 0ºC. After stirring for 24 hours at ambient temperature, the

resulting precipitate was removed by filtration. The filtrate was washed with 1N hydrochloric acid solution and evaporated in vacuo. The residue was dissolved in a mixture of acetic acid (80 ml) and water (80 ml), refluxed for 1 hour. Evaporation of the solvent gave a residue which was taken up into ethyl acetate. The solution was washed with saturated sodium bicarbonate and brine, dried over magnesium sulfate, and evaporated. The residue was chromatographed on silica gel eluting with a mixture of n-hexane and ethyl acetate (2:1, V/V) to give

( 2S)-1-allyloxycarbonyl-4-(2-oxopropyl)-2-(methoxymethyl)-pyrrolidine (12.4 g).

IR (Neat) : 1680-1700 cm^-1

NMR (CDCl₃, δ) : 2.04 (3H, s), 2.2-3.0 (5H, m),

3.34 (3H, s), 3.3-3.8 (3H, m) , 3.8-4.1 (2H, m), 4.58 (2H, m), 5.1-5.4 (2H, m) , 5.8-6.0 (1H, m)

Preparation 36-7)

To a solution of (3R,4R)-4-acetoxy-3-[(1R)-1-t-butyldimethylsilyloxyethyl]-2-oxoazetidine (15.4 g) and N-ethylpiperidine (7.2 ml) in dichloromethane (100 ml) was added trimethylsilyl trifluoromethanesulfonate (10.4 ml) at -60ºC under nitrogen, and the mixture was stirred at 0ºC for 30 minutes (Solution A). To a solution of

(2)-1-allyloxycarbonyl-4-(2-oxopropyl)-2-methoxymethylpyrrolidine (11.4 g) and N-ethylpiperidine (7.2 ml) in dichloromethane (100 ml) was added trimethylsilyl

trifluoromethanesulfonate (9.5 ml) at -60ºC under

nitrogen. The resulting mixture was stirred at 0ºC for 30 minutes. To this mixture was added dropwise the former mixture (Solution A) and trimethylsilyl

trifluoromethanesulfonate (3.0 ml) at 0°º, and the mixture was stirred for another 4 hours at 0ºC. The reaction mixture was taken up into a mixture of ethyl acetate and water and the mixture was stirred at ambient temperature for 2 hours. After adjusting pH to around 6.5 with aqueous sodium hydrogen carbonate, the organic layer was separated, washed with brine, dried over magnesium sulfate and evaporated in vacuo. The residue was chromatographed on silica gel eluting with a mixture of n-hexane and ethyl acetate (1:2, V/V) to give (3S,4R)-4-[3-{(2S)-1-allyloxycarbonyl-2-(methoxymethyl)pyrrolidin-4-yl}-2-oxopropyl]-3-[(1R)-1-t-butyldimethylsilyloxyethyl]-2-oxoazetidine (13.76 g).

Preparation 37-1)

Ethyl 1-benzylpiperidine-Δ^4,α-acetate was obtained in substantially the same manner as that of Preparation

36-3).

IR (Neat) : 1715, 1655 cm^-1

NMR (CDCl₃, δ) : 1.26 (3H, t, J=7Hz), 2.33 (2H, t,

J=6Hz), 2.53 (4H, t, J=6Hz), 2.99 (2H, t,

J=6Hz), 3.54 (2H, s), 4.12 (2H, q, J=7Hz), 5.63 (1H, s), 7.2-7.4 (5H, m) Preparation 37-2) To a solution of ethyl 1-benzylpiperidine-Δ^4,α- acetate (50 g) in ethanol (400 ml) was added palladium on carbon (9 g) and the mixture was stirred for 4 hours under hydrogen atmosphere. The catalyst was filtered off and the filtrate was concentrated in vacuo to give a residue. The residue was dissolved in a mixture of water (200 ml) and tetrahydrofuran (200 ml). To the mixture was added dropwise allyl chloroformate (22.4 ml) while adjusting pH to 8.0-9.0 with 30% aqueous sodium hydroxide at 0ºC.

After stirring for 2 hours at room temperature, the solvent was evaporated to give a residue. The residue was partitioned between ether and water. The ether layer wasconcentrated in vacuo to give 1-allyloxycarbonyl-4- (ethoxycarbonylmethyl)piperidine (50 g).

IR (Neat) : 3450, 1730, 1695 cm^-1

NMR (CDCl₃, δ) : 1.23 (3H, t, J=7Hz), 2.24 (2H, d, J=7Hz), 2.85 (2H, m) , 4.1-4.3 (4H, m) , 4.5-4.6 (2H, m), 5.1-5.4 (2H, m) , 5.8-6.0 (1H, m) Preparation 37-3)

1-Allyloxycarbonyl-4-(2-oxopropyl)piperidine was obtained in substantially the same manner as those of Preparations 1-7) and 36-6).

IR (Neat) : 1690 cm^-1

NMR (CDCl₃, δ) : 1.0-1.3 (2H, m) , 1.6-1.8 (2H, m),

1.9-2.1 (1H, m), 2.14 (3H, s), 2.38 (2H, d, J=7Hz), 2.6-2.9 (2H, m) , 4.0-4.2 (2H, m) ,

4.5-4.6 (2H, m), 5.1-5.4 (2H, m) , 5.8-6.0 (1H, m)

Preparation 37-4)

(3S,4R)-4-[3-(1-Allyloxycarbonylpiperidin-4-yl)-2-oxopropyl]-3-[(1R)-1-t-butyldimethylsilyloxyethyl]-2-oxoazetidine was obtained in substantially the same manner as that of Preparation 36-7). IR (Neat) : 3300, 1750, 1700 cm^-1

NMR (CDCl₃, δ) : 0.07 (6H, s), 0.86 (9H, s), 1.0-1.3 (5H, m), 2.37 (2H, d, J=7Hz), 2.4-3.0 (5H, m) , 3.8-4.0 (1H, m), 4.0-4.3 (3H, m) , 4.5-4.6 (2H, m), 5.1-5.4 (2H, m) , 5.8-6.0 (1H, m) , 6.09 (1H, br s)

Preparation 38-1)

To a solution of (2S,4R)-1-allyloxycarbonyl-2-hydroxymethyl-4-(2-hydroxypropyl)pyrrolidine (3.5 g) in dichloromethane (70 ml) at 0ºC were added triethylamine (2.4 ml), t-butyldimethylsilyl chloride (2.28 g) and

4-(dimethylamino)pyridine (70 mg). After stirring at room temperature for 12 hours, the solution was washed with lN-hydrochloric acid solution, saturated sodium

bicarbonate, and brine. The dried solvent was evaporated and then the obtained residue was chromatographed on silica gel eluting with a mixture of n-hexane and ethyl acetate (2:1, V/V) to give (2S,4R)-1-allyloxycarbonyl-2-t-butyldimethylsilyloxymethyl-4-(2-hydroxypropyl)-pyrrolidine (3.3 g).

IR (Neat) : 3400, 1690 cm^-1

NMR (CDCl₃, δ) : 0.87 (9H, s), 1.2-1.3 (3H, m),

1.4-1.8 (4H, m), 2.1-2.3 (2H, m) , 2.7-2.9 (1H, m), 3.7-4.0 (5H, m) , 4.5-4.6 (2H, m) , 5.1-5.4

(2H, m), 5.8-6.0 (1H, m)

Preparation 38-2)

( 2S,4R)-1-Allyloxycarbonyl-2-t-butyldimethylsilyloxymethyl-4-(2-oxopropyl)pyrrolidine was obtained in substantially the same manner as that of Preparation

36-6).

IR (Neat) : 1700 cm^-1

NMR (CDCl₃, δ) : 0.87 (9H, s), 1.5-1.7 (1H, m) , 2.15 ( 3H, s ) , 2. 2-2 . 8 ( 5H, m) , 3 . 6-4. 0 ( 4H, m) , 4. 5-4. 6 ( 2H, m) , 5 .1-5 . 4 ( 2H, m) , 5. 8-6. 0 ( 1H, m) Preparation 38-3)

(3S,4R)-4-[3-{(2S,4R)-1-allyloxycarbonyl-2-(t- butyldimethylsilyloxymethyl)pyrrolidin-4-yl}-2-oxopropyl]- 3-[(1R)-1-t-butyldimethylsilyloxyethyl]-2-oxoazetidine was obtained in substantially the same manner as that of

Preparation 36-7).

IR (Neat) : 3300, 1750, 1700 cm^-1

NMR (CDCl₃, δ) : 0.87 (18H, s), 1.22 (3H, d, J=6Hz), 3.6-4.0 (5H, m), 4.0-4.3 (1H, m) , 4.5-4.6 (2H, m), 5.1-5.4 (2H, m) , 5.8-6.0 (1H, m) , 6.02 (1H, br s)

Preparation 39-1)

1-Benzyloxycarbonyl-3-oxoazetidine (23 g) was

obtained in substantially the same manner as that of

Preparation 36-2). This substance was dissolved in toluene (200 ml). To this solution was added

1-(triphenylphosphoranylidene)butan-2-one (25 g), and the mixture was warmed at 90ºC. After stirring for 2 hours, evaporation of the solvent gave a residue, which was chromatographed on silica gel eluting with a mixture of n-hexane and ethyl acetate (1:1, V/V) to give

1-benzyloxycarbonylazetidine-Δ^3,α-2-oxobutane (14.5 g).

IR (Nujol) : 1700, 1645 cm^-1

NMR (CDCl₃, δ) : 1.08 (3H, t, J=7Hz), 2.48 (2H, q, J=7Hz), 4.6-4.7 (2H, m) , 4.9-5.0 (2H, m) , 5.16

(2H, s), 6.16 (3H, t, J=2Hz), 7.2-7.4 (5H, m)

Preparation 39-2)

(3S,4R)-4-[(1-Benzyloxycarbonylazetidine)-Δ^3,3-(1-methyl-2-oxopropyl]-3-[(1R)-1-t-butyldimethylsilyloxy- ethyl]-2-oxoazetidine was obtained in substantially the same manner as that of Preparation 36-7).

IR (CH₂Cl₂) : 1750, 1710, 1650 cm^-1

NMR (CDCl₃, δ) : 0.84 (9H, s), 1.1-1.3 (6H, m),

2.4-2.9 (2H, m) , 4.6-4.7 (2H, m) , 4.8-4.9 (2H, m), 5.23 (2H, s), 6.0, 6.05 (1H, each br s), 6.1-6.2 (1H, m), 7.2-7.4 (5H, m)

Preparation 39-3)

(3S,4R)-4-[3-(1-Allyloxycarbonylazetidin-3-yl)-1- methyl-2-oxopropyl]-3-[(1R)-1-t-butyldimethylsilyloxy- ethyl]-2-oxoazetidine was obtained in substantially the same manner as that of Preparation 37-2).

IR (CH₂Cl₂) : 1765, 1705 cm^-1

NMR (CDCl₃, δ) : 0.08 (6H, s), 0.80 (9H, s), 1.1-1.3

(6H, m), 2.5-3.0 (5H, m) , 4.4-4.5 (2H, m) , 5.0-5.3 (2H, m), 5.8-6.0 (1H, m) , 6.05 (1H, br s) Preparation 40-1)

To a solution of 1-t-butyloxycarbonyl-2-methoxycarbonyl-3-oxopyrrolidine (8 g) in a mixture of tetrahydrofuran (100 ml) and ethanol (80 ml) was added sodium borohydride (0.62 g) at -50ºC. After stirring for 30 minutes at the same temperature, the solvent was evaporated. The residue was poured into a mixture of ethyl acetate and water. The organic layer was washed in turn with 1N-hydrochloric acid, saturated sodium- bicarbonate, and brine. The dried solvent was evaporated, and resulting crude oil was dissolved in dimethylformamide (50 ml). To the solution were added t-butyldimethylsilyl chloride (6.4 g) and imidazole, and the mixture was stirred for 1 hour at ambient temperature. The mixture was poured into a mixture of ethyl acetate and water, and the organic layer was washed three times with water. The dried solvent was evaporated to give

1-t-butyloxycarbonyl-3-t-butyldimethylsilyloxy-2- methoxycarbonylpyrrolidine (11.4 g).

IR (CH₂Cl₂) : 1735, 1680 cm^-1

NMR (CDCl₃, δ) : 0.02-0.03 (6H, m) , 0.82 (9H, s),

1.23, 1.33 (9H, each s), 1.8-2.0 (2H, m),

3.2-3.4 (1H, m), 3.60 (3H, s), 3.4-3.7 (1H, m), 4.1-4.4 (1H, m), 4.4-4.6 (1H, m) Preparation 40-2)

To a solution of 1-t-butyloxycarbonyl-3-t-butyldimethylsilyloxy-2-methoxycarbonylpyrrolidine (26 g) in diethyl ether (300 ml) was added lithium aluminum hidride (4 g) at -30ºC. After stirring for 1.5 hours at the same temperature, to the mixture was added saturated aqueous potassium sodium tartrate solution. After

decantation, the solvent was evaporated, and then the obtained residue was chromatographed on silica gel eluting with a mixture of n-hexane and ethyl acetate to give

1-t-butyloxycarbonyl-3-t-butyldimethylsilyloxy-2- (hydroxymethyl)pyrrolidine (13.6 g).

IR (Neat) : 3350, 1660 cm^-1

NMR (CDCl₃, δ) : 0.03 (6H, s), 0.81 (9H, s), 1.37 (9H, s), 1.6-1.9 (2H, m), 3.2-3.8 (4H, m), 4.2-4.4 (2H, m)

Preparation 40-3)

To a stirred solution of 1-t-butyloxycarbonyl-3-t-butyldimethylsilyloxy-2-(hydroxymethyl)pyrrolidine ( 14.0 g) in tetrahydrofuran (140 ml) were added methyl iodide (10 ml) and sodium hydride (60% in oil, 2.0 g) at 0ºC. After stirring for 30 minutes at ambient temperature, the reaction mixture was taken up into a mixture of water (400 ml) and ethyl acetate (400 ml). The separated organic layer was washed in turn with hydrochloric acid (1N), aqueous sodium hydrogen carbonate, and brine, and dried over sodium sulfate. Evaporation of the solvent gave 1-t-butyloxycarbonyl-3-t-butyldimethylsilyloxy-2- (methoxymethyl)pyrrolidine (15.4 g).

IR (Neat) : 1695 cm^-1

NMR (CDCl₃, δ) : 0.02-0.04 (6H, m), 0.88 (9H, s), 1.46 (9H, s), 1.6-2.0 (2H, m) , 3.31 (3H, s), 3.2-3.8 (5H, m), 4.3-4.4 (1H, m) Preparation 40-4)

1-t-Butyloxycarbonyl-2-methoxymethyl-3-oxopyrrolidine was obtained in substantially the same manner as those of Preparations 36-1) and 36-2).

IR (Neat) : 1760, 1690 cm^-1

NMR (CDCl₃, δ) : 1.49 (9H, s), 2.5-2.6 (2H, m),

3.28 (3H, s), 3.5-4.1 (5H, m)

Preparation 40-5)

Ethyl 1-t-butyloxycarbonyl-2-(methoxymethyl)¬pyrrolidine-Δ^3,α-acetate was obtained in substantially the same manner as that of Preparation 36-3).

IR (Neat) : 1690 cm^-1

NMR (CDCl₃, δ) : 1.2-1.3 (3H, m), 1.47 (9H, s),

3.30, 3.33 (3H, each s), 3.4-3.8 (3H, m),

5.7-5.9 (1H, m)

Preparation 40-6)

1-t-Butyloxycarbonyl-3-ethoxycarbonylmethyl-2-(methoxymethyl)pyrrolidine was obtained in substantially the same manner as that of Preparation 36-4).

IR (Neat) : 1720, 1690 cm^-1

NMR (CDCl₃, δ) : 1.2-1.3 (3H, m) , 1.44 (9H, s),

3.28, 3.31 (3H, each s), 4.1-4.2 (2H, m) Preparation 40-7) 1-Allyloxycarbonyl-3-carboxymethyl-2-(methoxymethyl)pyrrolidine was obtained in substantially the same manner as that of Preparation 36-5).

IR (Neat) : 1680-1700 cm^-1

NMR (CDCl₃, δ) : 3.29, 3.34 (3H, each s), 4.0-4.1

(1H, s), 4.5-4.7 (2H, m), 5.1-5.4 (2H, m),

5.8-6.0 (1H, m)

Preparation 40-8)

1-Allyloxycarbonyl-3-(2-oxopropyl)-2- (methoxymethyl)pyrrolidine was obtained in substantially the same manner as that of Preparation 36-6).

IR (Neat) : 1690 cm^-1

NMR (CDCl₃, δ) : 2.14 (3H, s), 2.3-2.8 (3H, m),

3.2-3.6 (4H, m), 3.27-3.32 (3H, each s), 3.9-4.1

(1H, m), 4.5-4.6 (2H, m), 5.1-5.4 (2H, m),

5.8-6.0 (1H, m)

Preparation 40-9)

(3S,4R)-4-[3-{1-Allyloxycarbonyl-2-(methoxymethyl)pyrrolidin-3-yl}-2-oxopropyl]-3-[(1R)-1-t-butyldimethyl-silyloxyethyl]-2-oxoazetidine was obtained in

substantially the same manner as that of Preparation

36-7).

IR (Neat) : 3300, 1750, 1700 cm^-1

NMR (CDCl₃, δ) : 0.03 (6H, s) , 0.87 (9H, s), 1.21 (3H, d, J=6Hz), 3.2-3.7 (6H, m), 3.9-4.0 (1H, m), 4.0-4.2 (2H, m) , 4.5-4.6 (2H, m) , 5.1-5.4 (2H, m), 5.8-6.0 (1H, m), 6.0-6.2 (1H, m)

Preparation 41-1)

1-t-Butyloxycarbonyl-3-(3-oxo-1-butenyl)pyrrolidine was obtained in substantially the same manner as those of Preparations 36-2) and 36-3).

IR (Neat) : 1680, 1620 cm^-1 NMR ( CDCl₃ , δ ) : 1.74 ( 9H, s) , 1. 6-2. 3 ( 2H, m) , 2.26 ( 3H, s ) , 2. 8-3 . 7 ( 5H, m) , 6 . 14 ( 1H, d, J=16Hz ) , 6.71 ( 1H, dd, J=16Hz , 7. 6Hz ) Preparation 41-2)

1-t-Butyloxycarbonyl-3-(3-oxobutyl)pyrrolidine was obtained in substantially the same manner as that of

Preparation 36-4).

IR (Neat) : 1700, 1690 cm^-1

NMR (CDCl₃, δ) : 1.45 (9H, s), 2.15 (3H, s),

2.47 (2H, d, J=7Hz)

Preparation 41-3)

1-Allyloxycarbonyl-3-(3-oxobutyl)pyrrolidine was obtained in substantially the, same manner as that of

Preparation 36-5).

IR (Neat) : 1700, 1690 cm^-1

NMR (CDCl₃, δ) : 1.3-2.2 (5H, m) , 2.15 (3H, s),

2.48 (2H, t, J=7Hz), 2.93 (1H, dd, J=10Hz, 8Hz), 3.2-3.4 (1H, m) , 3.4-3.6 (2H, m) , 4.5-4.6 (2H, m), 5.1-5.4 (2H, m) , 5.8-6.0 (1H, m)

Preparation 41-4)

(3S,4R)-4-[4-(1-Allyloxycarbonylpyrrolidin-3-yl)-2-oxobutyl]-3-[(1R)-1-t-butyldimethylsilyloxyethyl]-2-oxoazetidine was obtained in substantially the same manner as that of Preparation 36-7).

IR (Neat) : 3250, 1750, 1690 cm^-1

NMR (CDCl₃, δ) : 0.06 (6H, s), 0.86 (9H, s), 1.20 (3H, d, J=6Hz), 2.47 (2H, d, J=8Hz), 2.5-3.0

(4H, m), 3.2-3.8 (3H, m) , 3.9-4.0 (1H, m) ,

4.1-4.2 (1H, m), 4.5-4.6 (2H, m) , 5.1-5.4 (2H, m), 5.8-6.0 (1H, m) , 6.08 (1H, br s) Preparation 42-1) To a stirred solution of methylthioacetonitrile (8.31 g) in dichloromethane (160 ml) was added by portions m-chloroperbenzoic acid (19.7 g) at 0ºC. After stirring at 0ºC for 30 minutes, the resulting precipitate was removed by filtration. To the filtrate was added

m-chloroperbenzoic acid (19.7 g) and the mixture was stirred at ambient temperature for 15 hours. The reaction mixture was diluted with ethyl acetate (350 ml), and the solution was washed with aqueous potassium carbonate (30%, 40 ml). The resulting precipitate was removed by

filtration and the filtrate was evaporated. The residue was recrystallized from dichloromethane to give

methylsulfonylacetonitrile (7.55 g).

ΪR (Nujol) : 2260, 1320, 1140 cm^-1

NMR (DMSO-d₆, δ) : 3.23 (3H, s) , 4.94 (2H, s)

Preparation 42-2)

Methyl methylsulfonylacetimidate (containing about 15% of starting nitrile) (4.87 g) was obtained from methylsulfonylacetonitrile (4.0 g) in substantially the same manner as that of Preparation 35-1).

NMR (CDCl₃, δ) : 3.00 (3H, s), 3.78 (3H, s),

3.90 (2H, s)

Preparation 43-1)

To a solution of ethyl difluoromethylthioacetate

(24.71 g) in ethanol (120 ml) was introduced ammonia gas at 45ºC for 6 hours. Evaporation of the solvent and trituration with diisopropyl ether gave

2- (difluoromethylthio) acetamide (15.83 g).

NMR (DMSO-d₆, δ) : 3.49 (2H, s), 7.35 (1H, t,

J=56Hz)

Preparation 43-2) To a solution of 2-(difluoromethylthio)acetamide (15.69 g) in pyridine (80 ml) was added trifluoroacetic anhydride (19.6 ml) at ambient temperature. Evaporation of the mixture gave a residue which was taken up into ethyl acetate (150 ml). The solution was washed in turn with hydrochloric acid (1N), aqueous sodium hydrogen carbonate and brine, and dried over magnesium sulfate. Evaporation of the solvent and chromatography on silica gel (200 ml) eluting with a mixture of n-hexane and ethyl acetate (95:5 - 4:1, V/V) gave

(difluoromethylthio)acetonitrile (12.0 g).

IR (Nujol) : 2250 cm^-1

NMR (CDCl₃, δ) : 3.59 (2H, s), 6.96 (1H, t, J=56Hz) Preparation 43-3)

Ethyl (difluoromethylthio)acetimidate hydrochloride was obtained in substantially the same manner as that of Preparation 35-1).

NMR (DMSO-d₆, δ) : 1.35 (3H, t, J=7Hz), 4.14 (2H, s), 4.50 (2H, q, J=7Hz), 7.54 (1H, t, J=56Hz)

Preparation 44-1)

(Methoxyacetylamino)acetonitrile was obtained in substantially the same manner as that of Preparation 16-1).

NMR (CDCl₃, δ) : 3.32 (3H, s), 3.89 (2H, s),

4.13 (2H, d, J=5.5Hz), 8.61 (1H, br s)

Preparation 44-2)

Ethyl (methoxymethylamino)acetimidate hydrochloride was obtained in substantially the same manner as that of Preparation 35-1).

NMR (DMSO-d₆, δ) : 1.33 (3H, t, J=7Hz), 3.33 (3H, s), 3.92 (2H, s), 4.21 (2H, d, J=5.5Hz), 4.51 (2H, q, J=7Hz), 8.58 (1H, br s)

To a stirred solution of allyl (5R,6S)-3-[(2S)-2,3- bis(allyloxycarbonylamino)propyl]-6-[(1R)-1-hydroxyethyl]- 7-oxo-1-azabicyclo[3.2.0]hept-2-ene-2-carboxylate (5.14 g) in a mixture of tetrahydrofuran (77 ml) and ethanol (26 ml) were added successively triphenylphosphine (1.13 g), dimedone (6.04 g), acetic acid (0.61 ml), and

tetrakis(triphenylphosphine)palladium(0) (1.00 g). After stirring at ambient temperature for 30 minutes, the reaction mixture was diluted with ethyl acetate (250 ml). The resulting precipitate was collected by filtration, washed in turn with ethyl acetate and dichloromethane, and taken up into cold phosphate buffer solution (pH 7, 250 ml). To this mixture was added by portions ethyl

acetimidate hydrochloride (7.39 g) at 2ºC while adjusting pH to 8.5 with aqueous potassium carbonate (30%), and the mixture was stirred for another 30 minutes at pH 8.5.

After adjusting pH to 7 with 1N hydrochloric acid, the reaction mixture was washed with a mixture of

tetrahydrofuran and ethyl acetate (3:7 V/V, 200 ml × 5), and evaporated in vacuo. The residue was chromatographed on nonionic adsorption resin, "Diaion HP-20" (Trademark, made by Mitsubishi Chemical Industries) (700 ml) eluting with a mixture of water and acetonitrile (100:0-97:3 V/V). The fractions containing the desired compound were

collected and lyophilized. The obtained powder was dissolved in a mixture of acetonitrile, water and ethanol, which was chromatographed on silica gel (50 ml) eluting with a mixture of acetonitrile and water (100:0-7:3 V/V) The fractions containing the desired compound were collected and lyophilized to give ( 5R,6S)-3-[{(4S)-2-methyl-2-imidazolin-4-yl}methyl]-6-[(1R)-1-hydroxyethyl]-7-oxo-1-azabicyclo[3.2.0]hept-2-ene-2-carboxylic acid (0.49 g).

IR (Nujol) : 3200, 1760 cm ^-1

NMR (D₂O, δ) : 1.28 (3H, d, J=6Hz), 2.20 (3H, s), 2.43-3.42 (5H, m), 3.45-4.80 (5H, m)

Example 2

(5R,6S)-3-[{{4R)-2-Methyl-2-imidazolin-4-yl}methyl]-6-[(1R)-1-hydroxyethyl]-7-oxo-1-azabicyclo[3.2.0]hept-2-ene-2-carboxylic acid was obtained in 23.3% yield in substantially the same method as that of Example 1.

IR (Nujol) : 3200, 1760 cm^-1

NMR (D₂O, δ) : 1.28 (3H, d, J=6Hz), 2.21 (3H, s), 2.88 (4H, d, J=8Hz), 3.32 (1H, dd, J=2, 6Hz), 3.42-4.75 (5H, m)

Example 3-1)

(5R,6S)-3-[(2-Methyl-2-imidazolin-4-yl)methyl]-6- [(1R)-1-hydroxyethyl]-7-oxo-1-azabicyclo[3.2.0]hept-2-ene-2-carboxylic acid was obtained in 38.8% yield in

substantially the same manner as that of Example 1.

IR (Nujol) : 3200, 1750 cm^-1

NMR (D₂O, δ) 1.25 (3H, d, J=6Hz), 2.05-2.30 (3H, m), 2.40-4.70 (10H, m)

Example 3-2)

(5R,6S)-3-[(2-Imidazolin-4-yl)methyl]-6-[(1R)-1-hydroxyethyl]-7-oxo-1-azabicyclo[3.2.0]hept-2-ene-2-carboxylic acid was obtained in 42.3% yield in

substantially the same method as that of Example 1 using benzyl formimidate hydrochloride instead of ethyl

acetimidate hydrochloride.

IR (Nujol) : 3250, 1750, 1630 cm ^-1

NMR (D₂O, δ) : 1.25 (3H, d, J=6Hz), 2.40-4.80 (10H, m), 7.70-8.20 (1H, m)

Example 4-1)

(5R,6S)-6-[(1R)-1-Hydroxyethyl]-3-[(2-imidazolin-1- yl)methyl]-7-oxo-1-azabicyclo[3.2.0]hept-2-ene-2-carboxylic acid (150 mg) was obtained from allyl

(5R,6S)-3-[N-allyloxycarbonyl-N-(2-allyloxycarbonylaminoethyl)aminomethyl]-6-[(1R)-1-hydroxyethyl]-7-oxo-1- azabicyclo[3.2.0]hept-2-ene-2-carboxylate (1.38 g) in substantially the same method as that of Example 1 using benzyl formimidate hydrochloride instead of ethyl

acetimidate hydrochloride.

IR (Nujol) : 3300, 1760, 1640 cm ^-1

NMR (D₂O, δ) 1.29 (3H, d, J=6Hz), 2.92 (2H, d, J=9Hz), 3.40 (1H, dd, J=3, 6Hz), 3.91 (4H, s), 4.03-4.70 (4H, m) , 8.15 (1H, s)

Example 4-2)

(5R,6S)-6-[(1R)-1-Hydroxyethyl]-3-[(2-methyl-2-imidazolin-1-yl)methyl]-7-oxo-1-azabicyclo[3.2.0]hept-2-ene-2-carboxylic acid was obtained in 15.0% yield in substantially the same mεthod as that of Example 4-1) using morpholine instead of dimedone and acetic acid.

IR (Nujol) : 3300, 1750 cm^-1

NMR (D₂O, δ) 1.27 (3H, d, J=6Hz), 2.24 (3H, s), 2.90 (2H, d, J=9Hz), 3.39 (1H, dd, J=3, 6Hz), 3.84 (4H, s), 4.00-4.70 (4H, m) Example 4-3)

(5R,6S)-3-[(2-Carbamoylmethyl-2-imidazolin-1-yl)- methyl]-6-[(1R)-1-hydroxyethyl]-7-oxo-1-azabicyclo[3.2.0]hept-2-ene-2-carboxylic acid was obtained in 8.4% yield in substantially the same method as that of Example 4-1) using ethyl carbamoylacetimidate hydrochloride instead of ethyl acetimidate hydrochloride.

IR (Nujol) : 3250, 1760, 1690 cm ^-1

NMR (D₂O, δ) : 1.25 (3H, d, J=6Hz), 2.90 (2H, d,

J=9Hz), 3.35 (1H, dd, J=3, 6Hz), 3.95 (4H, s), 3.90-4.85 (6H, m)

(5R,6S)-6-[(1R)-1-Hydroxyethyl]-3-[2-(1-methyl-3- pyridiniomethyl)-2-imidazolin-1-ylmethyl]-7-oxo-1- azabicyclo[3.2.0]hept-2-ene-2-carboxilic acid iodide was obtained in 13.1% yield in substantially the same method as that of Example 4-1) using ethyl 2-(l-methyl-3- pyridinio) acetimidate hydrochloride iodide instead of ethyl acetimidate hydrochloride.

IR (Nujol) : 3300, 1750 cm ^-1

NMR (D₂O, δ) 1.23 (3H, d, J=6Hz), 2.70-4.80 (13H, m), 4.30 (3H, s), 7.90-9.00 (4H, m) Example 5-1)

Allyl (5R,6S)-3-[(2S,4S)-1-allyloxycarbonyl-2- (N,N-dimethylcarbamoyl)pyrrolidin-4-ylmethyl]-6-[(IR)-1-t-butyldimethylsilyloxyethyl]-7-oxo-1-azabicyclo[3.2.0]-hept-2-ene-2-carboxylate was obtained in 64.7% yield in substantially the same manner as that of Preparation 1-10).

IR (Neat) : 1780, 1700, 1660 cm^-1

NMR (CDCl₃, δ) : 0.07 (6H, s), 0.85 (9H, s),

2.97 (3H, s), 3.05 (3H, s), 4.3-4.8 (5H, m) , 5.0-5.5 (4H, m), 5.8-6.0 (2H, m)

Example 5-2)

Allyl (5R,6S)-3-[(2S,4R)-1-allyloxycarbonyl-2-(N,N-dimethylcarbamoyl)pyrrolidin-4-ylmethyl]-6-[(lR)-1-t-butyldimethylsilyloxyethyl]-7-oxo-1-azabicyclo[3.2.0]hept-2-ene-2-carboxylatε was obtained quantitatively in

substantially the same manner as that of Preparation

1-10).

IR (Neat) : 1780, 1705, 1650 cm^-1

NMR (CDCl₃, δ) : 0.07 (6H, s), 0.90 (9H, s), 2.95 (3H, s), 3.08 (3H, s), 4.4-4.9 (5H, m), 5.1-5.6 (4H, m), 5.8-6.0 (2H, m)

Example 5-3)

Allyl (5R,6S)-3-(l-allyloxycarbonylpyrrolidin-3-ylmethyl)-6-[(1R)-1-t-butyldimethylsilyloxyethyl]-7-oxo-1-azabicyclo[3.2.0]hept-2-ene-2-carboxylate was obtained quantitatively in substantially the same manner as that of Preparation 1-10).

IR (Neat) : 1710, 1780 cm ^-1

NMR (CDCl₃, δ) 0.08 (6H, s), 0.91 (9H, s),

3.0-3.2 (2H, m), 3.3-3.9 (3H, m) , 4.5-4.9 (4H, m), 5.1-5.4 (4H, m) , 5.8-6.0 (2H, m)

Example 5-4)

Allyl (5R,6S)-3-[(2S)-1-allyloxycarbonylpyrrolidin-2-ylmethyl]-6-[(1R)-1-t-butyldimethylsilyloxyethyl]-7-oxo-1-azabicyclo[3.2.0]hept-2-ene-2-carboxylate was obtained in 80.9% yield in substantially thε same manner as that of Preparation 1-10).

IR (Neat) : 1785, 1700 cm ^-1 NMR (CDCl₃, δ) 0.03 (6H, s), 0.85 (9H, s),

4.4-4.8 (4H, m), 5.0-5.5 (4H, m),

5.8-6.1 (2H, m)

Example 5-5)

Allyl (5R,6S)-3-(1-allyloxycarbonylazetidin-3-ylmethyl)-6-[(1R)-1-t-butyldimethylsilyloxyethyl]-7-oxo-1- azabicyclo[3.2.0]hept-2-ene-2-carboxylate was obtained in 99.0% yield in substantially the same manner as that of Preparation 1-10).

IR (Neat) : 1780, 1710 cm ^-1

NMR (CDCl₃, δ) 0.03 (6H, s), 0.91 (9H, s),

2.8-3.1 (4H, m), 3.6-3.8 (2H, m) , 4.5-4.8 (4H, m), 5.2-5.5 (4H, m) , 5.8-6.1 (2H, m)

Example 6-1)

Allyl (5R,6S)-3-[(2S,4S)-1-allyloxycarbonyl-2- (N,N-dimethylcarbamoyl)pyrrolidin-4-ylmethyl]-6-[(1R)-1-hydroxyethyl]-7-oxo-1-azabicyclo[3.2.0]hept-2-ene-2-carboxylate was obtained in 81.9% yield in substantially the same manner as that of Preparation 1-11).

IR (Neat) 1780, 1700, 1650 cm^-1 NMR (CDCl₃, δ) : 2.96 (3H, s), 3.10 (3H, s),

4.4-4.8 (5H, m), 5.1-5.5 (4H, m),

5.7-6.0 (2H, m)

Example 6-2)

Allyl (5R,6S)-3-[(2S,4R)-1-allyloxycarbonyl-2-(N,N-dimethylcarbamoyl)pyrrolidin-4-ylmethyl]-6-[(1R)-1-hydroxyethyl]-7-oxo-1-azabicyclo[3.2.0]hept-2-ene-2-carboxylate was obtained in 20.7% yield in substantially the same manner as that of Preparation 1-11).

IR (Neat) : 1780, 1700, 1650 cm^-1

NMR (CDCl₃, δ) : 2.16 (3H, d, J=7Hz), 2.96 (3H, s), 3.07 (3H, s), 3.6-4.3 (3H, m), 4.40-4.9 (5H, m),

5.1-5.6 (4H, m), 5.7-6.0 (2H, m)

Example 6-3)

Allyl (5R,6S)-3-(1-allyloxycarbonylpyrrolidin-3-ylmethyl)-6-[(1R)-1-hydroxyethyl]-7-oxo-1-azabicyclo[3.2.0]hept-2-ene-2-carboxylate was obtained in 90.0% yield in substantially the same manner as that of

Preparation 1-11). IR (Neat) : 1780, 1710 cm^-1

NMR (CDCl₃, δ) : 3.2-3.7 (3H, m) , 4.5-4.9 (4H, m),

5.1-5.4 (4H, m), 5.8-6.0 (2H, m)

Example 6-4)

Allyl (5R,6S)-3-[(2S)-1-allyloxycarbonylpyrrolidin-2-ylmethyl]-6-[(1R)-1-hydroxyethyl]-7-oxo-1-azabicyclo-[3.2.0]hept-2-ene-2-carboxylate was obtained in 57.9% yield in substantially the same manner as that of

Preparation 1-11).

IR (Neat) : 3450, 1780 cm^-1

NMR (CDCl₃, δ) : 2.5-3.6 (7H, m), 4.4-4.8 (4H, m),

5.0-5.5 (4H, m), 5.8-6.1 (2H, m)

Example 6-5)

Allyl (5R,6S)-3-(1-allyloxycarbonylazetidin-3-ylmethyl)-6-[(1R)-1-hydroxyethyl]-7-oxo-1-azabicyclo-[3.2.0]hept-2-ene-2-carboxylate was obtained in 83.0% yield in substantially the same manner as that of

Preparation 1-11).

IR (CH₂Cl₂) : 1770, 1700 cm^-1 NMR (CDCl₃, δ) : 2.8-3.2 (4H, m) , 3.7-3.9 (2H, m), 4.4-4.9 (4H, m), 5.2-5.5 (4H, m) , 5.7-6.1 (2H, m)

Example 7-1)

(5R,6S)-3-[(2S,4S)-2-(N,N-Dimethylcarbamoyl)-pyrrolidin-4-ylmethyl]-6-[(1R)-1-hydroxyethyl]-7-oxo-1-azabicyclo[3.2.0]hept-2-ene-2-carboxylic acid was obtained in 22.8% yield in substantially the same manner as that of the former part of Example 1.

IR (Nujol) : 1750 cm^-1

NMR (D₂O, δ) : 1.26 (3H, d, J=6Hz), 2.99 (3H, s),

3.06 (3H, s)

Example 7-2)

(5R,6S)-3-[(2S,4R)-2-(N,N-Dimethylcarbamoyl)-pyrrolidin-4-ylmethyl]-6-[(1R)-1-hydroxyethyl]-7-oxo-1-azabicyclo[3.2.0]hept-2-ene-2-carboxylic acid was obtained in 26.0% yield in substantially the same manner as that of Example 7-1).

IR (Nujol) 1750 cm ^-1 NMR (D₂O, δ) 1.25 (3H, d, J=6Hz), 2.96 (3H, s), 3.03 (3H, s)

Example 7-3)

(5R,6S)-6-[(1R)-1-Hydroxyethyl]-7-oxo-3-(pyrrolidin-3-ylmethyl]-1-azabicyclo[3.2.0]hept-2-ene-2-carboxylic acid was obtained in 73.0% yield in substantially the same manner as that of Example 7-1).

IR (Nujol) : 1750 cm^-1

NMR (D₂O, δ) : 1.1-1.4 (3H, m)

Example 7-4)

(5R,6S)-6-[(1R)-1-Hydroxyethyl]-7-oxo-3-[(2S)-pyrrolidin-2-ylmethyl]-1-azabicyclo[3.2.0]hept-2-ene-2-carboxylic acid was obtained in 56.2% yield in

substantially the same manner as that of Example 7-1).

IR (Nujol) : 1770 cm^-1

NMR (D₂O, δ) : 1.28 (3H, d, J=6Hz), 1.5-2.4 (4H, m)

MS (M⁺ + 1) 281 Example 7-5)

(5R,6S) 3-(Azetidin-3-ylmethyl)-6-[(1R)-1-hydroxyethyl]-7-oxo-1-azabicyclo[3.2.0]hept-2-ene-2-carboxylic acid was obtained in 68.4% yield in

substantially the same manner as that of Example 7-1).

IR (Nujol) : 1740 cm^-1

NMR (D₂O, δ) 1.25 (3H, d, J=6Hz), 2.3-3.4 (5H, m)

Example 8-1)

(5R,6S)-3-[(2S,4R)-2-(N,N-Dimethylcarbamoyl)-1-formimidoylpyrrolidm-4-ylmethyl]-6-[(1R)-1-hydroxyethyl]-7-oxo-1-azabicyclo[3.2.0]hept-2-ene-2-carboxylic acid was obtained in 46.4% yield in substantially the same manner as that of Example 8-2).

IR (Nujol) : 1750 cm^-1

NMR (D₂O, δ) : 1.26 (3H, d, J=6Hz), 2.96 (3H, s),

3.04 (3H, s), 7.81 (1H, br s) Example 8-2)

To a solution of (5R,6S)-6-[(1R)-1-hydroxyethyl]-7-oxo-3-(pyrrolidin-3-ylmethyl)-1-azabicyclo[3.2.0]hept-2-ene-2-carboxylic acid (600 mg) was added benzyl

formimidate hydrochloride (0.73 g) at 0ºC while adjusting pH to 8.5 with 30% aqueous sodium hydroxide solution.

After stirring for 30 minutes at the same temperature, the solution was adjusted to pH 6.5 with 1N-hydrochloric acid, and washed with ethyl acetate, concentrated in vacuo. The residue was chromatographed on nonionic adsorption resin "Diaion HP-20" (Trademark, made by Mitsubishi Chemical Industries) eluting with aqueous isopropyl alcohol (3%). The fractions containing the desired compound were

collected and lyophilized to give (5R,6S)-3-[1-formimidoylpyrrolidin-3-ylmethyl]-6-[(1R)-1-hydroxyethyl]-7-oxo-1-azabicyclo[3.2.0]hept-2-ene-2-carboxylic acid (360 mg).

IR (Nujol) : 1750 cm^-1

NMR (D₂O, δ) : 1.27 (3H, d, J=6Hz), 7.89 (1H, br s)

Example 8-3)

(5R,6S)-3-[(2S)-1-Formimidoylpyrrolidin-2-ylmethyl]-6-[(1R)-1-hydroxyethyl]-7-oxo-1-azabicyclo[3.2.0]hept-2-ene-2-carboxylic acid was obtained in 61.5% yield in substantially the same manner as that of Example 8-2).

IR (Nujol) : 1760 cm^-1

NMR (D₂O, δ) 1.26 (3H, d, J=6Hz), 1.5-2.4 (4H, m),

7.7-7.9 (1H, m)

MS : (M⁺ + 1) 308

Example 8-4)

(5R,6S)-6-[(1R)-1-Hydroxyethyl]-7-oxo-3-[(2S)-1-(N-methylformimidoyl)pyrrolidin-2-ylmethyl]-1-azabicyclo-[3.2.0]hept-2-ene-2-carboxylic acid was obtained in 41.7% yield in substantially the same manner as that of Example 8-2).

IR (Nujol) : 1750 cm^-1

NMR (D₂O, δ) : 1.27 (3H, d, J=6Hz), 1.6-2.3 (4H, m),

3.03 (3H, s), 7.71 (1H, br s)

Example 8-5)

( 5R,6S)-3-(1-Formimidoylazetidin-3-ylmethyl)-6-[(1R)- 1-hydroxyethyl]-7-oxo-1-azabicyclo[3.2-0]hept-2-ene-2-carboxylic acid was obtained in 68.1% yield in

substantially the same manner as that of Example 8-2).

IR (Nujol) : 1740 cm^-1

NMR (D_nO, δ) 1.25 (3H, d, J=6Hz), 2.3-3.4 (5H, m),

7.69 (1H, br s)

Example 9

(5R,6S)-3-[{(4R)-2-Carbamoylmethyl-2-imidazolin-4-yl}methyl]-6-[(1R)-1-hydroxyethyl]-7-oxo-1-azabicyclo- [3.2.0]hept-2-ene-2-carboxylic acid (330 mg) was obtained in 23.4% yield in substantially the same method as that of Example 4-3).

IR (Nujol) : 3150, 1750, 1695, 1620, 1570 cm^-1

NMR (D₂O, δ) : 1.28 (3H, d, J=6Hz ) , 2.70-3.12 (4H, m), 3.31 (1H, dd, J=3.6Hz), 3.45-4.80 (7H, m)

Example 10

(5R,6S)-3-[{(4R)-2-(N,N-Dimethylcarbamoyl)methyl-2-imidazolin-4-yl}methyl]-6-[(1R)-1-hydroxyethyl]-7-oxo-1-azabicyclo[3.2.0]hept-2-ene-2-carboxylic acid was obtained in 30.5% yield in substantially the same method as that of Example 1 using ethyl dimethylcarbamoylacetimidate

hydrochloride instead of ethyl acetimidate hydrochloride.

IR (Nujol) : 3250, 1750, 1640, 1570 cm^-1

NMR (D₂O, δ) : 1.25 (3H, d, J=6Hz), 2.72-3.20 (10H, m), 3.32 (1H, dd, J=3.6Hz), 3.45-4.80 (7H, m)

(continued on the next page)

Example 11

(5R,6S)-6-[(1R)-1-Hydroxyethyl]-3-[[(4R)-2-{3-(1-pyridinio)propyl}-2-imidazolin-4-yl]methyl]-7-oxo-1- azabicyclo[3.2.0]hept-2-ene-2-carboxylic acid·chloride was obtained in 18.3% yield according to a similar manner to that of Example 1.

IR (Nujol) : 3250, 1750, 1570 cm^-1

NMR (D₂O, δ) : 1.26 (3H, d, J=6Hz), 2.00-3.05 (10H, m), 3.20-4.85 (6H, m) , 7.90-9.00 (5H, m)

Example 12

(5R,6S)-6-[(1R)-1-Hydroxyethyl]-3-[{(4R)-2-(3-pyridilmethyl)-2-imidazolin-4-yl}methyl]-7-oxo-1-azabicyclo[3.2.0]hept-2-ene-carboxylic acid was obtained in 27.2% yield according to a similar manner to that of Example 1.

IR (Nujol) : 3200, 1760, 1580 cm^-1

NMR (D₂O, δ) : 1.26 (3H, d, J=6Hz), 2.60-3.10 (4H, m), 3.20 (1H, dd, J=3Hz, 6Hz), 3.41-4.80 (5H, m), 4.00 (2H, s), 7.30-7.56 (1H, m), 7.68-7.90

(1H, m), 8.30-8.60 (2H, m)

Example 13

Using Amberlyst A-26 (C1-type, trademark, made by Rohm & Haas Co.) (25 ml) (eluting with water) instead of silica gel column chromatography, (5R,6S)-6-[(1R)-1-hydroxyethyl]-3-[[(4R)-2-{(1-methyl-3-pyridinio)methyl}-2-imidazolin-4-yl]methyl]-7-oxo-1-azabicyclo[3.2.0]hept-2-ene-2-carboxylic acid chloride was obtained in 28.2% yield according to a similar manner to that of Example 1.

IR (Nujol) : 3250, 1750, 1580 cm^-1

NMR (D₂O, δ) : 1.26 (3H, d, J=6Hz), 2.65-3.10 (4H, m), 3.31 (1H, dd, J=3Hz, 6Hz), 3.45-4.80 (7H, m), 4.40 (3H, s), 7.80-8.20 (1H, m) , 8.26-9.00 (3H, m) Example 14

( 5R,6S)-3-[{(4S)-2-Carbamoylmethyl-2-imidazolin-4-yl}methyl]-6-[(1R)-1-hydroxyethyl]-7-oxo-1-azabicyclo[3.2.0]hept-2-ene-2-carboxylic acid was obtained in 19.0% yield according to a similar manner to that of Example 9.

IR (Nujol) : 3150, 1750, 1690 cm^-1

NMR (D₂O, δ) : 1.26 (3H, d, J=6Hz), 2.50-3.50 (5H, m), 3.50-4.85 (5H, m) Example 15

(5R,6S)-3-[{(4R)-2-Carbamoylmethyl-2-imidazolin-4-yl}methyl]-6-[(1R)-1-hydroxyethyl]-7-oxo-1-azabicyclo[3.2.0]hept-2-ene-2-carboxylic acid (0.18 g) was dissolved in a mixture of phosphate buffer solution (6 ml)(pH 6.86) and acetonitrile (2 ml) and cooled to 0ºC. Acetic

anhydride (0.101 ml) was dropped into this solution. The pH of the reaction mixture was kept at around 8.5 in the course of reaction by adding aqueous potassium carbonate. After the addition of acetic anhydride the solution was stirred at pH 8.5 and at 0ºC for 1 hour, and then the pH was adjasted to 7 by 1N hydrochloric acid before

evaporation of the organic solvent under reduced pressure. The residue was subjected to column chromatography on silica gel (20 ml) (eluting with acetonitrile:water = 100:0 - 70:30, V/V) and the fraction containing the object compound were collected and lyophilized to give

(5R,6S)-3-[{(4R)-1(and

3-)-acetyl-2-carbamoylmethyl-2-imidazolin-4-yl}methyl]-6-[(1R)-1-hydroxyethyl]-7-oxo-1-azabicyclo[3.2.0]hept-2-ene-2-carboxylic acid (200 mg).

IR (Nujol) : 3300, 1750, 1650, 1580 cm^-1

NMR (D₂O, δ) : 1.26 (3H, d, J=6Hz), 1.80-2.30 (3H, m), 2.65-3.05 (4H, m) , 3.00-3.40 (1H, m) ,

3.45-4.85 (5H, m) Example 16

To a stirred solution of allyl (5R,6S)-3-[(2R)-2,3-bis(allyloxycarbonylamino)propyl]-6-[(1R)-1-hydroxyethyl]-7-oxo-1-azabicyclo[3.2.0]hept-2-ene-2-carboxylate (0.7 g) and triphenylphosphine (0.15 g) in a mixture of

tetrahydrofuran (9 ml) and ethanol (3 ml) were added successively acetic acid (0.5 ml) and

tetrakis(triphenylphosphine)palladium(0) (0.14 g) at 30ºC. After stirring at 30ºC for 5 minutes, to the mixture was added tri-n-butyltin hydride (1.6 ml). After stirring at ambient temperature for 15 minutes, the resulting

precipitate was filtered, washed with ethyl acetate, and taken up into phosphate buffer (pH 6.86, 50 ml). To the solution was added methyl methylsulfinylacetimidate (1.2 g) at ambient temperature while adjusting pH to around 8.5 with aqueous potassium carbonate (10%), and stirring was continued at pH 8.5 for another 2 hours. After adjusting pH to 7 with IN hydrochloric acid, the reaction mixture was concentrated to 3 ml, and diluted with acetonitrile (5 ml) and ethanol (2 ml). This solution was subjected to silica gel chromatography (100 ml) eluting with a mixture of acetonitrile and water (10:0 - 7:3, V/V).

Lyophilization of the eluate gave (5R,6S)-6-[(1R)-1-hydroxyethyl]-3-[{(4R)-2-(methylsulfinylmethyl)-2-imidazolin-4-yl}methyl]-7-oxo-1-azabicyclo[3.2.0Jhept-2-ene-2-carboxylic acid (222 mg).

IR (Nujol) : 3150, 1750, 1570 cm^-1

NMR (D₂O, δ) : 1.27 (3H, d, J=6Hz), 2.85 (3H, s), 2.65-3.10 (4H, m) , 3.31 (1H, dd, J=3.6Hz),

3.50-4.90 (5H, m)

The following compounds were obtained in

substantially the same manner as that of Example 1. Example 17 (5R,6S)-6-[(1R)-1-Hydroxyethyl]-3-[{(4R)-2-(1- methyltetrazol-5-ylthiomethyl)-2-imidazolin-4-yl}methyl]- 7-oxo-1-azabicyclo[3.2.0]hept-2-ene-2-carboxylic acid

IR (Nujol) : 3200, 1750, 1570 cm^-1

NMR (D₂O, δ) : 1.28 (3H, d, J=6Hz), 2.65-3.05 (4H, m), 3.32 (1H, dd, J=3Hz, 6Hz), 3.45-4.95 (7H, m), 4.05 (3H, s)

Example 18

(5R,6S)-6-[(1R)-1-Hydroxyethyl]-3-[{(4R)-2-(methyl-thiomethyl)-2-imidazolin-4-yl}methyl]-7-oxo-1-azabicyclo- [3.2.0]hept-2-ene-2-carboxylic acid

IR (Nujol) : 3250, 1750, 1570 cm^-1

NMR (D₂O, δ) : 1.26 (3H, d, J=6Hz), 2.16 (3H, s), 2.70-3.15 (4H, m), 3.32 (1H, dd, J=3Hz, 6Hz),

3.45-4.85 (5H, m), 3.53 (2H, s)

Example 19

(5R,6S)-3-[[(4R)-2-{(2-Carbamoyloxyethyl)thiomethyl}-2-imidazolin-4-yl]methyl]-6-[(IR)-1-hydroxyethyl]-7-oxo-1-azabicyclo[3.2.0]hept-2-ene-2-carboxylic acid

IR (Nujol) : 3150, 1750, 1700, 1570 cm^-1

NMR (D₂O, δ) : 1.28 (3H, d, J=6Hz), 2.60-3.10 (6H, m), 3.32 (1H dd, J=3, 6Hz), 3.40-4.80 (7H, m) , 3.66 (2H, s)

Example 20

(5R,6S)-3-[[(4R)-2-{(N,N-Dimethylcarbamoylmethylthio)methyl}-2-imidazolin-4-yl]methyl]-6-[(1R)-1- hydroxyethyl]-7-oxo-l-azabicyclo[3.2.0]hept-2-ene-2- carboxylic acid

IR (Nujol) : 3200, 1750, 1600 cm^-1

NMR (D₂O, δ) : 1.26 (3H, d, J=6Hz), 2.55-3.18 (10H, m), 3.30 (1H, dd, J=3, 6Hz), 3.40-4.80 (7H, m) , 3.55 (2H, m) Example 21

(5R,6S)-6-[(1R)-1-Hydroxyethyl]-3-[[(4R)-2-{(2-methoxyethylthio)methyl}-2-imidazolin-4-yl]methyl]-7-oxo-1-azabicyclo[3.2.0]hept-2-ene-2-carboxylic acid

IR (Nujol) : 3050, 1740, 1560 cm^-1

NMR (D₂O, δ) : 1.26 (3H, d, J=6Hz), 2.70-3.10 (6H, m), 3.24-3.48 (4H, m), 3.50-4.90 (9H, m)

Example 22

(5R,6S)-3-[[(4R)-2-(t-Butylthiomethyl)-2-imidazolin-4-yl]methyl]-6-[(1R)-1-hydroxyethyl]-7-oxo-l-azabicyclo[3.2.0]hept-2-ene-2-carboxylic acid

IR (Nujol) : 3200, 1760, 1580 cm^-1

NMR (D₂O, δ) : 1.28 (3H, d, J=6Hz), 1.34 (9H, s), 2.60-3.20 (4H, m) , 3.33 (1H, dd, J=3, 6Hz),

3.45-4.85 (5H, m), 3.65 (2H, s)

Example 23

(5R,6S)-3-[{(4R)-2-(Allylthiomethyl)-2-imidazolin-4-yl}methyl]-6-[(1R)-1-hydroxyethyl]-7-oxo-1-azabicyclo-[3.2.0]hept-2-ene-2-carboxylic acid

IR (Nujol) : 3150, 1750, 1580 cm^-1

NMR (D₂O, δ) : 1.29 (3H, d, J=6Hz), 2.60-4.80 (14H, m), 4.95-5.40 (2H, m) , 5.50-6.20 (1H, m)

Example 24

(5R,6S)-6-[(1R)-1-Hydroxyethyl]-3-[[(4R)-2-{(1-methylimidazol-2-yl)thiomethyl}-2-imidazolin-4-yl]-methyl]-7-oxo-1-azabicyclo[3.2.0]hept-2-ene-2-carboxylic acid

IR (Nujol) : 3150, 1750, 1580 cm^-1

NMR (D₂O, δ) : 1.27 (3H, d, J=6Hz), 2.60-3.10 (4H, m), 3.31 (1H, dd, J=3, 6Hz), 3.40-4.80 (7H, m), 3.76 (3H, s), 7.11 (1H, d, J=1.2Hz), 7.29 (1H, d, J=1.2Hz) Example 25

(5R,6S)-6-[(1R)-1-Hydroxyethyl]-3-[[(4R)-2-{(methanesulfonylamino)methyl}-2-imidazolin-4-yl]methyl]-7-oxo-1- azabicyclo[3.2.0]hept-2-ene-2-carboxylic acid

IR (Nujol) : 3150, 1750, 1570 cm^-1

NMR (D₂O, δ) : 1.25 (3H, d, J=6Hz), 2.74-3.05 (4H, m), 3.12 (3H, s), 3.32 (1H, dd, J=3Hz, 6Hz), 3.45-4.85 (5H, m), 4.24 (2H, s) Example 26

To a stirred solution of allyl (5R,6S)-3-[(2R)-2,3- bis(allyloxycarbonylamino)propyl]-6-[(1R)-1-hydroxyethyl]- 7-oxo-1-azabicyclo[3.2.0]hept-2-ene-2-carboxylate (1.20 g) and triphenylphosphine (0.11 g) in a mixture of

tetrahydrofuran (15 ml) and ethanol (5 ml) were added successively acetic acid (0.86 ml),

tetrakis (triphenylphosphine)palladium(0) (0.23 g) and tri-n-butyltin hydride (2.70 ml) at 30ºC. After stirring at ambient temperature for 15 minutes, the reaction mixture was diluted with ethyl acetate (20 ml) and the resulting precipitate was collected by filtration, washed rapidly with ethyl acetate, and taken up into a cold phosphate buffer solution (pH 7, 100 ml). To this

solution was added by portions at 0°C ethyl

(acetylamino)acetimidate hydrochloride (0.91 g) while adjusting pH to around 8.5 with aqueous potassium

carbonate (30%), and the mixture was stirred for another 30 minutes at pH 8.5. After adjusting pH to 7 with hydrochloric acid ( 1N) , the reaction mixture was washed with a mixture of tetrahydrofuran and ethyl acetate (3:7, V/V, 100 ml × 4) and evaporated in vacuo. The residue was chromatographed on nonionic adsorption resin, Diaion HP-20 (Trademark, made by Mitsubishi Chemical Industries) (120 ml) eluting with a mixture of water and acetonitrile

(100:0 - 9:1, V/V). After lyophilization, the obtained powder was dissolved in a mixture of water and acetonitrile, which was chromatographed on silica gel (50 ml) eluting with a mixture of acetonitrile and water (100:0 - 7:3, V/V).

The eluate was lyophilized to give (5R,6S)-3-[{(4R)-2- (acetylamino)methyl-2-imidazolin-4-yl}methyl]-6-[(1R)-1-hydroxyethyl]-7-oxo-1-azabicyclo[3.2.0]hept-2-ene-2-carboxylic acid (0.27 g).

IR (Nujol) : 3200, 1750, 1570 cm^-1

NMR (D₂O, δ) : 1.26 (3H, d, J=6Hz), 2.50 (3H, s),

2.70-3.10 (4H, m), 3.33 (1H, dd, J=3, 6Hz), 3.40-4.90 (5H, m) , 4.24 (2H, s)

Example 27

(5R,6S)-6-[(1R)-1-Hydroxyethyl]-3-[{(4R)-2-(ureidomethyl)-2-imidazolin-4-yl}methyl]-7-oxo-1-azabicyclo[3.2. 0]hept-2-ene-2-carboxylic acid was obtained in

substantially the same manner as that of Example 1.

IR (Nujol) : 3250, 1750, 1660, 1580 cm^-1

NMR (D₂O, δ) : 1.28 (3H, d, J=7Hz), 2.70-3.02 (4H, m), 3.33 (1H, dd, J=3, 6Hz), 3.42-4.80 (7H, m)

Example 28

( 5R,6S)-6-[(1R)-1-Hydroxyethyl]-3-[{(4R)-2-{(methoxy¬carbonylamino)methyl}-2-imidazolin-4-yl}methyl]-7-oxo-1- azabicyclo[3.2.0]hept-2-ene-2-carboxylic acid was obtained in substantially the same manner as that of Example 16.

IR (Nujol) : 3150, 1750, 1720, 1570 cm^-1

NMR (D₂O, δ) : 1.27 (3H, d, J=6Hz), 2.65-3.10 (4H, m), 3.32 (1H, dd, J=3, 6Hz), 3.45-4.90 (7H, m) ,

3.69 (3H, s)

The following compounds were obtained in

substantially the same manner as that of Example 1. Example 29

( 5R,6S)-6-[(1R)-1-Hydroxyethyl]-3-[{(4S)-2-methoxymethyl)-2-imidazolin-4-yl}methyl]-7-oxo-1-azabicyclo[3.2.0]hept-2-ene-2-carboxylic acid

IR (Nujol) : 3150, 1750, 1570 cm^-1

NMR (D₂O, δ) : 1.29 (3H, d, J=6Hz) , 2.50-3.23 (4H, m), 3.32 (1H, dd, J=3Hz, 6Hz), 3.42 (3H, s), 3.55-5.00 (5H, m) , 4.40 (2H, s) Example 30

(5R,6S)-3-[{(4R)-2-(2-Carbamoyloxyethyl)-2- imidazolin-4-yl}methyl]-6-[(1R)-1-hydroxyethyl]-7-oxo-1- azabicyclo[3.2.0]hept-2-ene-2-carboxylic acid

NMR (D₂O, δ) : 1.26 (3H, d, J=6Hz), 2.60-3.05 (6H, m), 3.31 (1H, dd, J=3Hz, 6Hz), 3.40-4.80 (7H, m)

Example 31

( 5R,6S)-6-[(1R)-1-Hydroxyethyl]-3-[{(4R)-2-(2-methoxyethyl)-2-imidazolin-4-yl}methyl]-7-oxo-1-azabicyclo[3.2.0]hept-2-ene-2-carboxylic acid

IR (Nujol) : 3200, 1750, 1570 cm^-1

NMR (D₂O, δ) : 1.25 (3H, d, J=6Hz), 2.60-3.03 (6H, m), 3.28 (1H, dd, J=3Hz, 6Hz), 3.35 (3H, s), 3.40-4.75 (7H, m)

Example 32

(5R,6S)-6-[(1R)-1-Hydroxyethyl]-3-[{(4R)-2-(hydroxymethyl)-2-imidazolin-4-yl}methyl]-7-oxo-1-azabicyclo[3.2.0]hept-2-ene-2-carboxylic acid

IR (Nujol) : 3200, 1750, 1590 cm^-1

NMR (D₂O, δ) : 1.29 (3H, d, J=6Hz), 2.70-3.03 (4H, m), 3.33 (1H, dd, J=3Hz, 6Hz), 3.45-4.85 (5H, m), 4.52 (2H, s) Example 33 (5R,6S)-6-[(1R)-1-Hydroxyethyl]-3-[{(4R)-2-(2-hydroxyethyl)-2-imidazolin-4-yl}methyl]-7moxo-1- azabicyclo[3.2.0]hept-2-ene-2-carboxylic acid

IR (Nujol) : 3200, 1750, 1580 cm^-1

NMR (D₂O, δ) : 1.28 (3H, d, J=6Hz), 2.50-3.10 (6H, m), 3.32 (1H, dd, J=3Hz, 6Hz), 3.43-4.85 (7H, m)

Example 34

(5R,6S)-3-[{(4R)-2-(Carbamoyloxymethyl)-2-imidazolin-4-yl}methyl]-6-[(1R)-1-hydroxyethyl]-7-oxo-1-azabicyclo-[3.2.0]hept-2-ene-2-carboxylic acid

IR (Nujol) : 3250, 1750, 1620, 1580 cm^-1

NMR (D₂O, δ) : 1.28 (3H, d, J=6Hz), 2.75-3.10 (4H, m), 3.32 (1H, dd, J=3Hz, 6Hz), 3.45-4.80 (5H, m), 4.95 (2H, s)

Example 35

(5R,6S)-6-[(1R)-l-Hydroxyethyl]-3-[{(4R)-2-(N-ethyl- carbamoyloxymethyl)-2-imidazolin-4-yl}methyl]-7-oxo-1- azabicyclo[3.2.0]hept-2-ene-2-carboxylic acid

IR (Nujol) : 3200, 1750, 1730, 1580 cm^-1

NMR (D₂O, δ) : 1.10 (3H, t, J=7Hz), 1.28 (3H, d,

J=6Hz), 2.75-3.09 (4H, m), 3.16 (2H, q, J=7Hz), 3.33 (1H, dd, J=3Hz, 6Hz), 3.45-4.85 (5H, m) , 4.94 (2H, s)

Example 36

( 5R,6S)-3-[{(5S)-2-Methoxymethyl-1-methy1-2- imidazolin-5-yl}methyl]-6-[(1R)-1-hydroxyethyl]-7-oxo-1-azabicyclo[3.2.0]hept-2-ene-2-carboxylic acid

IR (Nujol) : 3300, 1750

NMR (D₂O, δ) : 1.27 (3H, d, J=6.3Hz), 2.76-3.13 (4H, m), 3.06 (3H, s), 3.23-4.30 (5H, m) , 3.48 (3H, s), 4.47 (2H, s)

Example 37 (5R,6S)-6-[(1R)-1-Hydroxyethyl]-3-[{(4R)-2-(methoxymethyl)-2-imidazolin-4-yl}methyl]-7-oxo-1-azabicyclo- [3.2.0]hept-2-ene-2-carboxylic acid

IR (Nujol) : 3200, 1750, 1570 cm^-1

NMR (D₂O, δ) : 1.27 (3H, d, J=6Hz), 2.70-3.10 (4H, m), 3.31 (1H, dd, J=3Hz, 6Hz), 3.43 (3H, s), 3.47-4.60 (5H, m) , 4.41 (2H, s)

Example 38

To a stirred solution of allyl (5R,6S)-3-[2-[N- (allyloxycarbonyl) -N-{3-(allyloxycarbonylamino)propyl}- amino]methyl-6-[(1R)-1-hydroxyethyl]-7-oxo-1-azabicyclo[3.2.0]hept-2-ene-2-carboxylate and allyl (5R,6S)-3-[2- [N-(allyloxycarbonyl)-N-{3-(N-allyloxycarbonyl-N-methylamino)propyl}amino]methyl]-6-[(1R)-1-hydroxyethyl]--7- oxo-1-azabicyclo[3.2.0]hept-2-ene-2-carboxylate (total 17.47 g) in a mixture of tetrahydrofuran (210 ml) and ethanol (70 ml) were added successively triphenylphosphine (3.70 g), dimedone (19.78 g), acetic acid (2.02 ml), and tetrakis(triphenylphosphine)palladium(0) ( 3.26 g). After stirring at ambient temperature for 30 minutes, the resulting precipitate was collected by filtration, washed in turn with ethyl acetate and dichloromethane, and taken up into a cold phosphate buffer solution (pH 7, 400 ml). To this mixture was added by portions ethyl acetimidate hydrochloride (13.08 g) at 2ºC while adjusting pH to 8.5 with aqueous potassium carbonate (30%), and the mixture was stirred for another 30 minutes at pH 8.5. After adjusting pH to 7 with lN-aqueous hydrochloric acid, the reaction mixture was washed with a mixture of

tetrahydrofuran and ethyl acetate (3:7, V/V, 300 ml x 5), and evaporated in vacuo. The residue was chromatographed on nonionic adsorption resin, Diaion HP-20 (Trademark, made by Mitsubishi Chemical Industries) (2 ℓ) eluting with a mixture of water and acetonitrile (100:0-96:4, V/V). The fraction containing mainly ( 5R,6S)-6-[(IR)-1-hydroxyethyl]-3-[(2-methyl-1,4,5,6-tetrahydropyrimidin-1-yl)-methyl]-7-oxo-1-azabicyclo[3.2.0]hept-2-ene-2-carboxylic acid (hereinafter referred to as Compound A) and mainly

(5R,6S)-6-[(1R)-1-hydroxyethyl]-3-[(2,3-dimethyl-1,4,5,6- tetrahydro-1-pyrimidinio)methyl]-7-oxo-1-azabicyclo- [3.2- .0]hept-2-ene-2-carboxylate (hereinafter referred to as Compound B) were collected separately and each fraction was lyophilized to give a powder. The powder (1.3 g) containing mainly Compound A was dissolved in a mixture of acetonitrile and water, which was chromatographed on silica gel (200 ml) eluting with a mixture of acetonitrile and water (100:0 - 70:30, V/V). The fractions containing Compound A and Compound B were collected separately. The powder (1.7 g) containing mainly Compound B were separated in substantially the same manner as stated above to give the fractions containing Compound A and Compound B. The eluates containing Compound A were combined and

lyophilized to give (5R,6S)-6-[(1R)-1-hydroxyethyl]-3- [(2-methyl-1,4,5,6-tetrahydropyrimidin-1-yl)methyl]-7-oxo-1-azabicyclo[3.2.0]hept-2-ene-2-carboxylic acid (0.89 g). The eluates containing Compound B were combined and lyophilized to give (5R,6S)-6-[(1R)-1-hydroxyethyl]-3-[(2,3-dimethyl-1,4,5,6-tetrahydro-l-pyrimidinio)methyl]-7-oxo-1-azabicyclo[3.2.0]hept-2-ene-2-carboxylate ( 0.22 g).

Compound A :

IR (Nujol) : 1760 cm^-1

NMR (D₂O, δ) : 1.27 (3H, d, J=6Hz), 1.79-2.17 (2H, m), 2.24 (3H, s), 2.90 (2H, d, J=9Hz), 3.21-3.56 (5H, m) , 4.00-4.96 (4H, m) Compound B : IR (Nujol) : 1750 cm^-1

NMR (D₂O, δ) : 1.26 (3H, d, J=6Hz), 1.80-2.15 (2H, m), 2.28 (3H, s), 2.88 (2H, d, J=9Hz), 3.17 (3H, s), 3.25-3.58 (5H, m), 4.00-4.93 (4H, m)

Example 39

6-[(1R)-1-Hydroxyethyl]-3-[(1,4,5,6-tetrahydropyrimidin-1-yl)methyl]-7-oxo-1-azabicyclo- [3.2.0]hept-2-ene-2-carboxylic acid (hereinafter referred to as Compound C and 6-[(1R)-1-hydroxyethyl]-3-[(3-methyl-1,4,5,6-tetrahydro-1-pyrimidinio)methyl]-7- oxo-1-azabicyclo[3.2.0]hept-2-ene-2-carboxylate

(hereinafter referred to as Compound D) were obtained in substantially the same manner as that of Example 38.

Compound C :

IR (Nujol) : 3200-2500, 1750 cm^-1

NMR (D₂O, δ) : 1.27 (3H, d, J=6Hz) , 1.84-2.22 (2H, m), 2.86 (2H, d, J=9Hz), 3.21-3.53 (5H, m), 4.03-4.79 (4H, m), 7.95 (1H, s)

Compound D :

NMR (D₂O, δ) : 1.27 (3H, d, J=6Hz), 1.87-2.24 (2H, m), 2.86 (2H, d, J=9Hz), 3.10-3.49 (5H, m), 3.19 (3H, s), 4.00-4.82 (4H, m), 7.96 (1H, s)

The following compounds were obtained in

substantially the same manner as that of Example 1.

Example 40

(5R,6S)-6-[(1R)-1-Hydroxyethyl]-3-[2-(2-methyl-2-imidazolin-1-yl)ethyl]-7-oxo-1-azabicyclo[3.2.0]hept-2-ene-2-carboxylic acid

NMR (D₂O, δ) : 1.27 (3H, d, J=6Hz), 2.25 (3H, s), 2.67-3.90 (11H, m) , 3.98-3.37 (2H, m) Example 41

(5R,6S)-6-[(1R)-1-Hydroxyethyl]-3-[2-{2,3-dimethyl-1- (2-imidazolinio)}ethyl]-7-oxo-1-azabicyclo[3.2.0]hept- 2-ene-2-carboxylate

NMR (D₂O, δ) : 1.27 (3H, d, J=6Hz), 2.15 (3H, s),

2.75-3.66 (11H, m) , 3.78 (3H, s), 3.93-4.26 (2H, m)

Example 42

(5R,6S)-6-[(1R)-l-Hydroxyethyl]-3-[{3-(2-hydroxy-ethyl)-1-(2-imidazolinio)}methyl]-7-oxo-1-azabicyclo- [3.2.0]hept-2-ene-2-carboxylate

IR (Nujol) : 3600-2500, 1750 cm^-1

NMR (D₂O, δ) : 1.26 (3H, d, J=6Hz), 2.89 (2H, d,

J=9Hz), 3.39 (1H, dd, J=3Hz, 6Hz), 3.45-4.76

(12H, m), 8.13 (1H, s)

Example 43-1)

To a solution of (3S,4R)-4-[3-{(2S)-1-allyloxy-carbonyl-2-(methoxymethyl)pyrrolidin-4-yl}-2-oxopropyl]-3-[(1R)-1-t-bιityldimethylsilyloxyethyl]-2-oxoazetidine (13.76 g), and triethylamine (7.9 ml) in dichloromethane (100 ml) was added allyl oxalyl chloride (5.2 g) at -20ºC under nitrogen, and the resulting mixture was stirred for 30 minutes at -20ºC. The reaction mixture was taken up into a mixture of water and ethyl acetate. The organic layer was separated, washed in turn with water, aqueous sodium hydrogen carbonate and brine, dried over magnesium sulfate, and evaporated. The residue was dissolved in xylene (75 ml), and triethyl phosphite (14.8 ml) was added. The resulting mixture was heated to 90ºC for 15 hours under nitrogen, and heated to 130ºC for 2 hours under nitrogen. Evaporation of the solvent gave a residue which was chromatographed on silica gel eluting with a mixture of n-hexane and ethyl acetate (4:1, V/V) to give allyl (5R,6S)-3-[(2S)-1-allyloxycarbonyl-2-(methoxy- methyl)pyrrolidin-4-yl]methyl-6-[(1R)-1-t-butyldimethylsilyloxyethyl]-7-oxo-1-azabicyclo[3.2.0]hept-2-ene-2- carboxylate (15.2 g) .

IR (Neat) : 1760, 1690 cm^-1

NMR (CDCl₃, δ) : 0.08 (6H, s), 0.89 (9H, s), 2.4-2.8 (4H, m), 2.8-3.1 (1H, m), 3.26 (3H, s), 4.4-4.8 (4H, m), 5.0-5.6 (4H, m), 5.8-6.0 (2H, m) The following compounds were obtained in

substantially the same manner as that of Example 43-1).

Example 43-2)

Allyl (5R,6S)-3-[1-allyloxycarbonylpiperidin-4-yl]-methyl-6-[(1R)-1-t-butyldimethylsilyloxyethyl]-7-oxo-1-azabicyclo[3.2.0]hept-2-ene-2-carboxylate

IR (Neat) : 1780, 1700 cm^-1

NMR (CDCl₃, δ) : 0.03 (6H, s), 0.90 (9H, s), 2.4-2.9 (5H, m), 3.0-3.1 (1H, m), 4.0-4.3 (4H, m) ,

4.5-4.8 (4H, m), 5.1-5.5 (4H, m), 5.8-6.0 (2H, m)

Example 43-3)

Allyl (5R,6S)-3-[(2S,4S)-1-allyloxycarbonyl-2-(t-butyldimethylsilyloxymethyl)pyrrolidin-4-yl]methyl-6-

[(1R)-1-t-butyldimethylsilyloxyethyl]-7-oxo-1-azabicyclo-[3.2.0]hept-2-ene-2-carboxylate

IR (Neat) : 1775, 1700 cm^-1

NMR (CDCl₃, δ) : 0.84 (18H, s), 3.6-4.0 (4H, m),

4.5-4.8 (4H, m) , 5.1-5.5 (4H, m), 5.8-6.0 (2H, m)

Example 43-4)

Allyl (5R,6S)-3-(1-allyloxycarbonylazetidin-3-yl)- methyl-6-[(1R)-1-t-butyldimethylsilyloxyethyl]-4-methyl- 7-oxo-1-azabicyclo[3.2.0]hept-2-ene-2-carboxylate

IR (Neat) : 1770, 1700 cm^-1

NMR (CDCl₃, δ) : 0.06 (6H, s), 0.81 (9H, s), 1.1-1.3 (6H, m), 2.5-2.8 (1H, m) , 2.9-3.1 (2H, m),

3.2-3.4 (1H, m), 3.5-3.7 (3H, m), 4.4-4.8 (4H, m), 5.0-5.4 (4H, m), 5.8-6.0 (2H, m)

Example 43-5)

Allyl (5R,6S)-3-[1-allyloxycarbonyl-2- (methoxvmethyl)pyrrolidin-3-yl]methyl-6-[(1R)-1-t-butyldimethylsilyloxyethyl]-7-oxo-1-azabicyclo[3.2.0]-hept-2-ene-2-carboxylate

IR (Neat) : 1780, 1700 cm^-1

NMR (CDCl₃, δ) : 0.03 (6H, s), 0.88 (9H, s), 2.7-2.9

(2H, m), 3.29, 3.33 (3H, each s), 4.5-4.8 (4H, m), 5.1-5.5 (4H, m) , 5.8-6.0 (2H, m)

Example 43-6)

Allyl (5R,6S)-3-[2-(1-allyloxycarbonylpyrrolidin-3-yl)ethyl]-6-[(1R)-1-t-butyldimethylsilyloxyethyl]-7-oxo-1-azabicyclo[3.2.0]hept-2-ene-2-carboxylate

IR (Neat) : 1770, 1700 cm^-1

NMR (CDCl₃, δ) : 0.06 (6H, s), 0.86 (9H, s), 2.4-2.6 (1H, m), 2.6-3.0 (3H, m) , 3.1-3.7 (3H, m) ,

4.4-4.8 (4H, m), 5.0-5.4 (4H, m) , 5.8-6.0 (2H, m)

Example 44-1)

To a solution of allyl (5R,6S)-3-[(2S)-1-allyloxy¬carbonyl-2-(methoxymethyl)pyrrolidin-4-yl]methyl-6-[(1R)-1-t-butyldimethylsilyloxyethyl]-7-oxo-1-azabicyclo[3.2.0]-hept-2-ene-2-carboxylate (15.2 g) in tetrahydrofuran (70 ml) were added acetic acid (4.6 ml) and tetrabutylammonium fluoride (30 ml, 70% solution in tetrahydrofuran) at 0ºC under nitrogen. After standing at ambient temperature for 7 hours and at 5ºC for 14 hours, the reaction mixture was taken up into a mixture of ethyl acetate and water. After adjusting pH to around 7 with aqueous sodium hydrogen carbonate, the organic layer was separated, washed in turn with water and brine, and dried over magnesium sulfate. Removal of the solvent gave a residue, which was

chromatographed on silica gel eluting with a mixture of n-hexane and ethyl acetate (1:2, V/V) to give allyl

( 5R,6S)-3-[(2S)-1-allyloxycarbonyl-2-(methoxymethyl)-pyrrolidin-4-yl]methyl-6-[(1R)-1-hydroxyethyl]-7-oxo-1-azabicyclo[3.2.0]hept-2-ene-2-carboxylate (10 g).

IR (Neat) : 3400, 1770, 1700 cm^-1

NMR (CDCl₃, δ) : 2.7-2.9 (2H, m), 3.1-3.2 (1H, m), 3.34 (3H, s), 4.5-4.8 (4H, m), 5.1-5.6 (4H, m),

5.8-6.0 (2H, m)

The following compounds were obtained in

substantially the same manner as that of Example 44-1).

Example 44-2)

Allyl (5R,6S)-3-(1-allyloxycarbonylpiperidin-4-yl)-methγl-6-[(1R)-1-hydroxyethyl]-7-oxo-1-azabicyclo[3.2.0]-hept-2-ene-2-carboxylate

IR (Neat) : 3400, 1770, 1690 cm^-1

NMR (CDCl₃, δ) : 2.3-2.9 (5H, m), 3.0-3.2 (1H, m), 4.0-4.3 (4H, m), 4.5-4.9 (4H, m), 5.1-5.5 (4H, m), 5.8-6.0 (2H, m) Example 44-3)

Allyl (5R,6S)-3-[(2S,4S)-1-allyloxycarbonyl-2-(hydroxymethyl)pyrrolidin-4-yl]methyl-6-[(1R)-1-hydroxyethyl]-7-oxo-1-azabicyclo[3.2.0]hept-2-ene-2-carboxylate

IR (Neat) : 3400, 1770, 1700 cm^-1 NMR (CDCl₃, δ) : 2.5-3.0 (4H, m), 3.15 (1H, dd,

J=3Hz, 7Hz), 4.5-4.9 (4H, m), 5.1-5.5 (4H, m), 5.8-6.0 (2H, m) Example 44-4)

Allyl (5R,6S)-3-(1-allyloxycarbonylazetidin-3-yl)-methyl-4-methyl-6-[(1R)-1-hydroxyethyl3-7-oxo-1-azabicycl[3.2.0]hept-2-ene-2-carboxylate

IR (CH₂Cl₂) : 3400, 1760, 1700 cm^-1

NMR (CDCl₃, δ) : 2.4-2.6 (1H, m) , 2.6-2.8 (1H, m),

3.0-3.2 (2H, m), 3.2-3.4 (1H, m), 3.6-3.8 (3H, m), 4.5-4.8 (4H, m), 5.2-5.5 (4H, m) , 5.8-6.0 (2H, m) Example 44-5)

Allyl ( 5R,6S)-3-[1-allyloxycarbonyl-2-(methoxymethyl)pyrrolidin-3-yl]methyl-6-[(1R)-1-hydroxyethyl]-7-oxo-1-azabicyclo[3.2.0]hept-2-ene-2-carboxylate

IR (Neat) : 3400, 1770, 1700 cm^-1

NMR (CDCl₃, δ) : 2.6-2.9 (3H, m), 3.28, 3.32 (3H, each s), 3.2-3.7 (4H, m) , 4.5-4.9 (4H, m), 5.1-5.5 (4H, m), 5.8-6.0 (2H, m)

Example 44-6)

Allyl (5R,6S)-3-[2-(1-allyloxycarbonylpyrrolidin-3-yl)ethyl]-6-[(1R)-1-hydroxyethyl]-7-oxo-1-azabicyclo-[3.2.0]hept-2-ene-2-carboxylate

IR (Neat) : 3400, 1770, 1700 cm^-1

NMR (CDCl₃, δ) : 2.8-3.0 (3H, m), 3.1-3.2 (1H, m), 3.2-3.4 (1H, m) , 3.4-3.6 (2H, m), 4.5-4.8 (4H, m), 5.1-5.6 (4H, m) , 5.8-6.0 (2H, m)

Example 45-1)

To a solution of allyl (5R,6S)-3-[(2S)-1-allyloxy-carbonyl-2-(methoxymethyl)pyrrolidin-4-yl]methyl-6-[(1R)- 1-hydroxyethyl]-7-oxo-1-azabicyclo[3.2.0]hept-2-ene-2- carboxylate (7.2 g) in a mixture of tetrahydrofuran (70 ml) and ethanol (30 ml) were added successively

triphenylphosphine (0.8 g) , 5,5-dimethyl-1,3-cyclohexanedione (dimedone) (4.6 g), and

tetrakis (triphenylphosphine)palladium(0) ( 1.8 g).

Stirring at ambient temperature for 1 hour gave a

precipitate, which was collected by filtration, and washed with tetrahydrofuran. The solid was chromatographed on silica gel eluting with a mixture of acetonitrile and water (7:3, V/V) to give (5R,6S)-6-[(1R)-1-hydroxyethyl]-7-oxo-3-[(2S)-2-(methoxymethyl)pyrrolidin-4-yl]methyl-1-azabicyclo[3.2.0]hept-2-ene-2-carboxylic acid (1.6 g).

IR (Nujol) : 1750 cm^-1

NMR (D₂O, δ) : 1.28 (3H, d, J=6.5Hz), 3.41 (3H, s)

MS : 325 (M⁺+l)

This product was separated by HPLC eluting with a mixture of water and acetonitrile (9:1, V/V) to give two fractions (First fraction and Second fraction).

First fraction :

(5R,6S)-6-[(1R)-1-Hydroxyethyl]-3-[(2S,4S)-2-(methoxymethyl)pyrrolidin-4-yl]methyl-7-oxo-1-azabicyclo-[3.2.0]hept-2-ene-2-carboxylic acid

IR (Nujol) : 1750 cm^-1

NMR (D₂O, δ) : 1.28 (3H, d, J=6.5Hz), 1.3-1.6 (1H, m), 2.2-2.4 (1H, m) , 2.4-3.1 (5H, m) , 3.41 (3H, s), 3.3-4.0 (6H, m), 4.0-4.3 (2H, m)

Second fraction :

(5R,6S)-6-[(IR)-1-Hydroxyethyl]-3-[(2S,4R)-2-(methoxymethyl)pyrrolidin-4-yl]methyl-7-oxo-1- azabicyclo[3.2.0]hept-2-ene-2-carboxylic acid

IR (Nujol) : 1750 cm^-1 NMR (D₂O, δ ) : 1.28 ( 3H, d, J=6.5Hz ) , 1. 8-2. 0 ( 2H, m) , 3 . 41 ( 3H, s ) , 3 . 9-4. 3 ( 3H, m)

The following compounds were obtained in

substantially the same manner as that of Example 45-1).

Example 45-2)

(5R,6S)-6-[(1R)-1-Hydroxyethyl]-3-(piperidin-4-yl)-methyl-7-oxo-1-azabicyclo[3.2.0]hept-2-ene-2-carboxylic acid

IR (Nujol) : 1750 cm^-1

NMR (D₂O, δ) : 1.28 (3H, d, J=6Hz), 1.2-1.6 (2H, m), 1.8-2.0 (3H, m), 2.3-2.5 (1H, m) , 2.6-2.8 (1H, m), 2.8-3.1 (4H, m) , 3.3-3.6 (3H, m) , 4.1-4.4 (2H, m)

Example 45-3)

(5R,6S)-6-[(lR)-1-Hydroxyethyl]-3-[(2S,4S)-2-(hydroxymethyl)pyrrolidin-4-ylmethyl]-7-oxo-1-azabicyclo[3.2.0]hept-2-ene-2-carboxylic acid

IR (Nujol) : 1760 cm^-1

NMR (D₂O, δ) : 1.28 (3H, d, J=6Hz), 1.4-1.6 (1H, m), 2.2-2.4 (1H, m), 2.5-3.0 (5H, m) , 3.3-3.6 (2H, m), 3.7-3.9 (3H, m) , 4.0-4.3 (2H, m)

Example 45-4)

( 5R,6S)-3-(Azetidin-3-ylmethyl)-6-[(1R)-1-hydroxyethyl]-4-methyl-7-oxo-1-azabicyclo[3.2.0]hept-2- ene-2-carboxylic acid

IR (Nujol) : 1755 cm^-1

NMR (D₂O, δ) : 1.22 (3H, d, J=7Hz), 1.28 (3H, d,

J=6Hz), 2.5-2.7 (1H, m), 3.0-3.4 (2H, m),

3.7-4.4 (3H, m) Example 45-5) (5R,6S)-6-[(1R)-1-Hydroxyethyl]-3-[2-(methoxymethyl)pyrrolidin-3-ylmethyl]-7-oxo-1-azabicyclo[3.2.0]-hept-2-ene-2-carboxylic acid

IR (Nujol) : 1740 cm^-1

NMR (D₂O, δ) : 1.28 (3H, d, J=6Hz), 2.8-3.0 (2H, m),

3.2-4.0 (6H, m), 3.40 (3H, s), 4.1-4.3 (2H, m)

Example 45-6)

(5R,6S)-6-[(1R)-1-Hydroxyethyl]-3-[2-(pyrrolidin-3-yl)ethyl]-7-oxo-1-azabicyclo[3.2.0]hept-2-ene-2-carboxylicacid

IR (Nujol) : 1750 cm^-1

NMR (D₂O, δ) : 1.28 (3H, d, J=6.5Hz), 1.3-1.8 (3H, m), 2.1-2.5 (2H, m) , 2.8-2.9 (2H, m) , 3.1-3.6 (4H, m), 4.0-4.3 (2H, m)

Example 46-1)

To a solution of (5R,6S)-6-[(1R)-1-hydroxyethyl]-7-oxo-3-[piperidin-4-yl]methyl-1-azabicyclo[3.2.0]hept-2-ene-2-carboxylic acid (600 mg) was added benzyl

formimidate hydrochloride (1.0 g) at 0ºC while adjusting pH to 8.5 with 30% aqueous sodium hydroxide solution.

After 30 minutes at the same temperature, the solution was adjusted to pH 6.5 with IN-hydrochloric acid, washed with ethyl acetate, and concentrated in vacuo. The residue was chromatographed on nonionic adsorption resin Diaion HP-20 (Trademark, made by Mitsubishi Chemical Industries) eluting with aqueous acetonitrile (3%). The fractions containing the desired compound were collected and

lyophilized to give (5R,6S)-3-(N-formimidoylpiperidin-4-ylmethyl)-6-[(1R)-1-hydroxyethyl]-7-oxo-1-azabicyclo-[3.2.0]hept-2-ene-2-carboxylic acid (220 mg).

IR (Nujol) : 1750, 1710 cm^-1

NMR (D₂O, δ) 1.28 (3H, d, J=6Hz), 1.2-1.6 (2H, m) 1.7-2.0 ( 3H, m) , 2.8-3.0 ( 1H, m) , 3.1-3.2 ( 1H, m) , 3.3-3.6 ( 2H, m) , 3.7-4.0 ( 2H, m) , 4.1-4.3 ( 2H, m)

Example 46-2 )

(5R,6S)-3-[2-(N-Formimidoylpyrrolidin-3-yl)ethyl]-6- [(1R)-1-hydroxyethyl]-7-oxo-1-azabicyclo[3.2.0]heρt-2-ene-2-carboxylic acid was obtained in substantially the same manner as that of Preparation 46-1).

IR (Nujol) : 1750, 1700 cm^-1

NMR (D₂O, δ) : 1.28 (3H, d, J=6.4Hz), 3.3-3.9 (5H, m), 4.1-4.3 (2H, m)

Example 47

To a solution of (5R,6S)-6-[(1R)-1-hydroxyethyl]-7-oxo-3-(piperidin-4-yl)methyl-1-azabicyclo[3.2.0]hept-2-ene-2-carboxylic acid (500 mg) in a mixture of water (20 ml) and dioxane (8 ml) was added dimethyl sulfonate (2 ml) at 0°C while adjusting pH to 9.0 with 1N-aqueous sodium hydroxide solution. After 1 hour at the same temperature, the solution was adjusted to pH 6.8 with IN-hydrochloric acid, washed with ethyl acetate, and concentrated in vacuo. The residue was chromatographed on nonionic adsorption resin Diaion HP-20 (Trademark, made by

Mitsubishi Chemical Industries) eluting with aqueous acetonitrile (3%). The fractions containing the desired compound were collected and lyophilized to give

( 5R, 6S)-3-(1,1-dimethyl-4-piperidinio)methyl-6-[(1R)-1-hydroxyethyl]-7-oxo-1-azabicyclo[3.2.0]hept-2-ene-2-carboxylate (150 mg).

IR (Nujol) : 1750 cm^-1

NMR (D₂O, δ) : 1.28 (3H, d, J=6Hz), 2.3-2.5 (2H, m), 2.6-2.8 (1H, m), 2.8-2.9 (2H, m), 3.08 (3H, s), 3.14 (3H, s), 3.1-3.6 (5H, m), 4.0-4.4 (2H, m) The following compounds were obtained in

substantially the same manner as that of Example 16.

Example 48

(5R,6S)-6-[(1R)-1-Hydroxyethyl]-3-[{(4R)-2- (methanesulfonylmethyl)-2-imidazolin-4-yl}methyl]-7-oxo-1-azabicyclo[3.2.0]hept-2-ene-2-carboxylic acid

IR (Nujol) : 3250, 1750, 1560 cm^-1

NMR (D₂O, δ) : 1.26 (3H, d, J=6Hz), 2.80-3.15 (4H, m), 3.27 (3H, s), 3.15-4.80 (6H, m)

Example 49

( 5R,6S)-3-[{(4R)-2-(Difluoromethylthiomethyl)-2-imidazolin-4-yl}methyl]-6-[(1R)-1-hydroxyethyl]-7-oxo-1-azabicyclo[3.2.0]hept-2-ene-2-carboxylic acid

NMR (D₂O, δ) : 1.28 (3H, d, J=6Hz), 2.80-3.06 (4H, m), 3.34 (1H, dd, J=3Hz, 6Hz), 3.90-4.25 (4H, m), 4.50-4.70 (1H, m), 7.15 (1H, t, J=55Hz) Example 50

( 5R,6S)-6-[(1R)-1-Hydroxyethyl]-3-[{(4R)-2-(methoxyacetylaminomethyl)-2-imidazolin-4-yl}methyl]-7-oxo-1-azabicyclo[3.2.0]hept-2-ene-2-carboxylic acid

IR (Nujol) : 3150, 1750, 1670, 1580 cm^-1

NMR (D₂O, δ) : 1.26 (3H, d, J=6Hz), 2.65-3.10 (4H, m), 3.20-4.80 (6H, m) , 3.42 (3H, s), 4.05 (2H, s), 4.31 (2H, s)

Claims

C LA I M

1. A compound of the formula :

in which R¹ is carboxy, protected carboxy or

carboxylato,

R² is hydroxy(lower)alkyl or protected

hydroxy(lower)alkyl,

R³ is azetidinyl, pyrrolidinyl.

imidazolinyl, tetrahydropyrimidinyl, piperidyl, 1,1-di(lower)alkylpiperidinio, 3-(lower)alkyl-1-(2- imidazolinio), 3-[hydroxy(lower)- alkyl]-1-(2-imidazolinio) or

3-(lower)alkyl-1,4,5,6-tetrahydro-1- pyrimidinio;

wherein each heterocyclic group may be substituted by one or more suitable substituent(s),

R¹⁰ is hydrogen or lower alkyl, and

A is lower alkylene,

provided that

when R³ is lower alkylpyrrolidinyl, then R¹⁰ is hydrogen,

or a pharmaceutically acceptable salt thereof.

2. The compound of Claim 1, wherein R³ is azetidinyl, pyrrolidinyl, imidazolinyl, tetrahydropyrimidinyl, piperidyl, 1,1-di(lower)alkyl- pyperidinio, 3-(lower)alkyl-1-(2-imidazolinio), 3-[hydroxy(lower)alkyl]-1-(2-imidazolinio) or 3-(lower)alkyl-1,4,5,6-tetrahydro-1-pyrimidinio; wherein the ring carbon atom(s) may be

substituted by the group consisting of lower alkyl, pyridyl(lower)alkyl, carbamoyl, mono(or di) (lower)alkylcarbamoyl, carbamoyl(lower)alkyl, mono(or di) (lower)alkylcarbamoyl(lower)alkyl, hydroxy(lower)alkyl, lower alkoxy(lower)alkyl, carbamoyloxy(lower)alkyl, mono(or di) ( lower)- alkylcarbamoyloxy(lower) alkyl, lower alkylthio- (lower) alkyl, lower alkylsulfinyl(lower)alkyl, lower alkenylthio(lower)alkyl, lower

alkoxy(lower)alkylthio(lower)alkyl,

carbamoyloxy(lower)alkylthio(lower)alkyl, mono(or di) ( lower)alkylcarbamoyl(lower)- alkylthio(lower)alkyl, lower

alkylsulfonyl(lower)alkyl, mono(or di or tri)- halo(lower)alkylthio(lower)alkyl, lower

alkoxy(lower)alkanoylamino(lower)alkyl, lower alkanoylamino(lower)alkyl, ureido(lower)alkyl, lower alkylsulfonylamino(lower)alkyl, lower alkoxycarbonylamino(lower)alkyl,

(1-pyridinio) (lower)alkyl,

1-(lower)alkylpyridinio(lower)alkyl and

N-containing heterocyclicthio(lower)alkyl wherein said heterocyclic group may be

substituted by lower alkyl, and the ring

nitrogen atom(s) may be substituted by the group consisting of lower alkyl, lower alkanimidoyl, N-(lower)alkyl(lower)alkanimidoyl and

imino-protective group. The compound of Claim 2, wherein

R¹ is carboxy, esterified carboxy or carboxylato.R² is hydroxy(lower)alkyl, acyloxy(lower)alkyl,

C₆-C₁₀ ar(lower)alkyloχy(lower)alkyl or

trisubstituted silyloxy(lower)alkyl;

R³ is azetidinyl, pyrrolidinyl, imidazolinyl,

tetrahydropyrimidinyl, piperidyl,

1,1-di(lower)alkylpiperidinio,

3-(lower)alkyl-1-(2-imidazolinio),

3-[hydroxy(lower)alkyl]-1-(2-imidazolinio) or

3-(lower)alkyl-1,4,5,6-tetrahydro-1-pyrimidinio; wherein the ring carbon atom(s) may be

substituted by one or two substituent(s)

selected from the group consisting of lower alkyl; pyridyl(lower)alkyl; carbamoyl; mono(or di) (lower)alkylcarbamoyl; carbamoyl(lower)alkyl; mono(or di) (lower)alkylcarbamoyl(lower)alkyl; hydroxy(lower)alkyl; lower alkoxy(lower)alkyl; carbamoyloxy(lower)alkyl; mono(or

di) (lower)alkylcarbamoyloxy(lower)alkyl; lower alkylthio(lower)alkyl; lower

alkylsulfinyl(lower)alkyl; lower

alkenylthio(lower)alkyl; lower alkoxy(lower)- alkylthio(lower)alkyl; carbamoyloxy(lower)- alkylthio(lower)alkyl; mono(or

di) (lower)alkylcarbamoyl(lower)alkylthio(lower)- alkyl; lower alkylsulfonyl(lower)alkyl;

dihalo(lower)alkylthio(lower)alkyl; lower alkoxy(lower)alkanoylamino(lower)alkyl; lower alkanoylamino(lower)alkyl, ureido(lower)alkyl; lower alkylsulfonylamino(lower)alkyl; lower alkoxycarbonylamino(lower)alkyl;

(1-pyridinio) (lower)alkyl;

1-(lower)alkylpyridinio(lower)alkyl; and a group consisting of pyrrolyl, pyrrolinyl, imidazolyl. pyrazolyl, pyridyl, pyridyl N-oxide, pyrimidyl , pyrazinyl , pyridazinyl , triazolyl , tetrazolyl and dihydrotriazinyl, each of which may be substituted by lower alkyl; and the ring nitrogen atom(s) may be substituted by the group consisting of lower alkyl, lower alkanimidoyl, N-(lower) alkyl(lower)alkanimidoyl and acyl.

4. The compound of Claim 3, wherein

R¹ is carboxy, lower alkenyloxycarbonyl, aryl(or

nitroaryl) (lower)alkoxycarbonyl or carboxylato, R² is hydroxy(lower)alkyl, aryl(or nitroaryl)(lower)- alkoxycarbonyloxy(lower)alkyl, lower

alkenyloxycarbonyloxy(lower)alkyl or

[tri(lower)alkylsilyloxy] (lower)alkyl, R³ is azetidinyl, pyrrolidinyl, imidazolinyl,

tetrahydropyrimidinyl, piperidyl, 1,1-di(lower)- alkylpiperidinio, 3-(lower)alkyl-1-(2-imidazolinio), 3-[hydroxy(lower)alkyl]-1-(2-imida- zolinio) or 3-(lower)alkyl-l,4,5,6-tetrahydro-1- pyrimidinio; wherein the ring carbon atom(s) may be substituted by the group consisting of lower alkyl, pyridyl(lower)alkyl, carbamoyl, mono(or di) (lower)alkylcarbamoyl, carbamoyl( lower)alkyl, mono(or di) (lower)alkylcarbamoyl(lower)alkyl, hydroxy(lower)alkyl, lower alkoxy(lower)alkyl, carbamoyloxy(lower)alkyl, mono(or di) ( lower)- alkylcarbamoyloxy(lower)alkyl, lower

alkylthio(lower)alkyl, lower

alkylsulfinyl(lower)alkyl, lower

alkenylthio(lower)alkyl, lower

alkoxy(lower)alkylthio( lower)alkyl,

carbamoyloxy(lower)alkylthio(lower)alkyl, mono(or di) (lower)alkylcarbamoyl(lower)- alkylthio(lower)alkyl, lower alkylsulfonyl(lower)alkyl,

dihalo(lower)alkylthio(lower)alkyl, lower alkoxy(lower)alkanoylamino(lower)alkyl, lower alkanoylamino(lower)alkyl, ureido(lower)alkyl, lower alkylsulfonylamino(lower)alkyl, lower alkoxycarbonylamino(lower)alkyl,

( 1-pyridinio) (lower)alkyl,

1-(lower)alkylpyridinio(lower)alkyl, lower alkylimidazolyl and lower alkyltetrazolyl, and the ring nitrogen atom(s) may be substituted by the group consisting of lower alkyl, lower alkanimidoyl, N-(lower)alkyl(lower)alkanimidoyl, lower alkanoyl, lower alkenyloxycarbonyl and aryl(or nitroaryl) (lower)alkoxycarbonyl.

5. The compound of Claim 4, wherein

R¹ is carboxy or carboxylato,

R² is hydroxy(lower)alkyl,

R³ is azetidinyl, pyrrolidinyl, imidazolinyl,

tetrahydropyrimidinyl, piperidyl, 1,1-di(lower)- alkylpiperidinio,

3-(lower)alkyl-1-(2-imidazolinio),

3-[hydroxy(lower)alkyl]-1-(2-imidazolinio) or 3-(lower)alkyl-1,4,5,6-tetrahydro-1-pyrimidinio; wherein the ring carbon atom(s) may be substituted by one or two substituent(s) selected from the group consisting of lower alkyl, pyridyl(lower)alkyl, carbamoyl, mono(or di) (lower)alkylcarbamoyl, carbamoyl(lower)alkyl, mono(or di) (lower)alkylcarbamoyl(lower)alkyl, hydroxy(lower)alkyl, lower alkoxy(lower)alkyl, carbamoyloxy(lower)alkyl, mono(or di) (lower) alkylcarbamoyloxy(lower)alkyl, lower

alkylthio(lower)alkyl, lower

alkylsulfinyl(lower)alkyl, lower alkenylthio(lower)alkyl, lower

alkoxy(lower)alkylthio(lower)alkyl,

carbamoyloxy(lower)alkylthio(lower)alkyl, mono(or di) (lower)alkylcarbamoyl(lower)- alkylthio(lower)alkyl, lower

alkylsulfonyl(lower)alkyl,

halo(lower)alkylthio(lower)alkyl, lower

(1-pyridinio)(lower)alkyl,

1-(lower)alkylpyridinio(lower)alkyl, lower alkylimidazolyl and lower alkyltetrazolyl, and the ring nitrogen atom(s) may be substituted by the group consisting of lower alkyl, lower alkanimidoyl, N-(lower)alkyl(lower)alkanimidoyl or lower alkanoyl. The compound of Claim 5, wherein

R³ is imidazolinyl, lower alkylimidazolinyl,

[carbamoyl(lower)alkyl]imidazolinyl,

[N,N-di(lower)alkylcarbamoyl(lower)alkyl]- imidazolinyl, [pyridyl(lower)alkyl]- imidazolinyl, azetidinyl,

1-(lower)alkanimidoylazetidinyl, pyrrolidinyl, 1- (lower)alkanimidoylpyrrolidinyl,

1-[N-(lower)alkyl(lower)alkanimidoyl]- pyrrolidinyl, [N,N-di(lower)alkylcarbamoyl]- pyrrolidinyl, 1-(lower)alkanimidoyl- [N,N-di(lower)alkylcarbamoyl]pyrrolidinyl,

[hydroxy(lower)alkyl]imidazolinyl, [lower alkoxy(lower)alkyl]imidazolinyl,

[carbamoyloxy(lower)alkyl]imidazolinyl, [mono or di (lower)alkylcarbamoyloxy(lower)alkyl]- imidazolinyl, [lower alkylthio(lower)alkyl]-imidazolinyl, [lower alkylsulfinyl(lower)alkyl]-imidazolinyl, [lower alkenylthio(lower)alkyl]-imidazolinyl, [lower alkoxy(lower)alkylthio- (lower)alkyl]imidazolinyl [carbamoyloxy(lower)- alkylthio(lower)alkyl]imidazolinyl, [mono or di(lower)alkylcarbamoyl(lower)alkylthio(lower)-alkyl]imidazolinyl, [lower alkylimidazolylthio-(lower)alkyl]imidazolinyl, [lower

alkyltetrazolylthio(lower)alkyl]imidazolinyl,

[lower alkanoylamino(lower)alkyl]imidazolinyl, [ureido(lower)alkyl]imidazolinyl, [lower

alkylsulfonylamino(lower)alkyl]imidazolinyl,

[lower alkoxycarbonylamino(lower)alkyl]-imidazolinyl, [lower alkyl] [lower alkoxy]- [lower alkoxy(lower)alkyl]imidazolinyl, N-[lower alkanoyl] [carbamoyl(lower)alkyl]imidazolinyl, tetrahydropyrimidinyl, (lower

alkyl)tetrahydropyrimidinyl, [pyridinio(lower)-alkyl]imidazolinyl, [1-(lower)alkylpyridinio- (lower)alkyl]imidazolinyl, 3-[hydroxy(lower)-alkyl]-1-(2-imidazolinio),

2 ,3-di(lower)alkyl-1-(2-imidazolinio),

3-(lower)alkyl-1,4,5,

6-tetrahydro-1-pyrimidinio optionally substituted by lower alkyl,

1,1-di(lower)alkylpiperidinio, [lower

alkylsulfonyl(lower)alkyl]imidazolinyl,

[dihalo(lower)alkylthio(lower)alkyl]imidazolinyl, [lower alkoxy(lower)alkanoylamino(lower)alkyl]-imidazolinyl, piperidyl, [lower

alkoxy(lower)alkyl]pyrrolidinyl,

[hydroxy(lower)alkyl]pyrrolidinyl or 1-(lower alkanimidoyl)piperidyl.

7. The compound of Claim 6, wherein

R² is hydroxy(C₁-C₄)alkyl,

R³ is imidazolinyl, 2-(C₁-C₄)alkyl-2-imidazolinyl, 2-carbamoyl(C₁-C₄)alkyl-2-imidazolinyl, 2-[N,N-di(C₁-C₄)alkylcarbamoyl(C₁-C₄)alkyl]-2- imidazolinyl, 2-{3-pyridyl(C₁-C₄)alkyl}-2- imidazolinyl, azetidmyl,

1-(C₁-C₄)alkanimidoylazetidinyl, pyrrolidinyl, 1-(C₁-C₄)alkanimidoylpyrrolidinyl,

1-[N- (C₁-C₄)alkyl(C₁-C₄)alkanimidoyl]pyrrolidinyl, [N,N-di(C₁-C₄)alkylcarbamoyl]pyrrolidinyl, 1- (C₁-C₄)alkanimidoyl-2-[N,N-di(C₁-C₄)- alkylcarbamoyl]pyrrolidinyl,

[hydroxy(C₁-C₄)alkyl]-2-imidazolinyl, [C₁-C₄ alkoxy(C₁-C₄)alkyl]-2-imidazolinyl,

[carbamoyloxy(C₁-C₄)alkyl]-2-imidazolinyl,

N-(C₁-C₄)alkylcarbamoyloxy(C₁-C₄)alkyl-2- imidazolinyl, [C₁-C₄ alkylthio(C₁-C₄)alkyl]-2- imidazolinyl, [C₁-C₄ alkylsulfinyl(C₁-C₄)- alkyl]-2- imidazolinyl, [C₂-C₄

alkenylthio(C₁-C₄)alkyl]-2-imidazolinyl, [C₁-C₄ alkoxy(C₁-C₄)alkylthio(C₁-C₄)alkyl]-2- imidazolinyl, [carbamoyloxy(C₁-C₄)alkylthio- (C₁-C₄)alkyl]-2-imidazolinyl,

[N,N-di(C₁-C₄)alkylcarbamoyl(C₁-C₄)alkylthio- ( C₁-C₄) alkyl]-2-imidazolinyl, [C₁-C₄

alkylimidazolylthio(C₁-C₄)alkyl]-2-imidazolinyl, [C₁-C₄ alkyltetrazolylthio(C₁-C₄)alkyl]-2- imidazolinyl, [C₁-C₄ alkanoylamino(C₁-C₄)alkyl]- 2-imidazolinyl, [ureido(C₁-C₄)alkyl]-2- imidazolinyl, [C₁-C₄ alkylsulfonylamino(C₁-C₄)- alkyl]-2-imidazolinyl, [C₁-C₄ alkoxycarbonylamino(C₁-C₄)alkyl]-2-imidazolinyl, N-[C₁-C₄ alkyl]-[C₁-C₄ alkoxy(C₁-C₄)alkyl]-2- imidazolinyl, N-[C₁-C₄ alkanoyl]-[carbamoyl- (C₁-C₄) alkyl]-2-imiazolinyl,

1,4,5,6-tetrahydropyrimidinyl, (C₁-C₄ alkyl)- 1,4,5,6-tetrahydropyrimidinyl,

[(1-pyridinio) (C₁-C₄)alkyl]-2-imidazolinyl,

[1-(C₁ -C₄)alkylpyridinio{C₁-C₄)alkyl]-2- imidazolinyl, 3-[hydroxy(C₁-C₄)alkyl]-1-(2- imidazolinio), [C₁-C₄

alkylsulfonyl(C₁-C₄)alkyl]- 2-imidazolinyl,

[dihalo(C₁-C₄)alkylthio(C₁-C₄)- alkyl]-2-imidazolinyl, [C₁-C₄ alkoxy(C₁-C₄)- alkanoylamino(C₁-C₄)alkyl]-2-imidazolinyl, 2,3-di(C₁-C₄)alkyl-1-(2-imidazolinio), 3-(C₁-C₄)alkyl-1,4,5,6-tetrahydro-1-pyrimidinio optionally substituted by C₁-C₄ alkyl, 1,1-di(C₁-C₄)alkylpiperidinio, piperidyl, [C₁-C₄ alkoxy(C₁-C₄)alkyl]pyrrolidinyl,

[hydroxy(C₁-C₄)alkyl]pyrrolidinyl or 1-(C₁-C₄ alkanimidoyl)piperidyl, and

A is C₁-C₄ alkylene.

8. The compound of Claim 7, wherein

R² is 1-hydroxyethyl,

R³ is 2-imidazolin-1(or 4)-yl,

2-methyl-2-imidazolin-1(or 4)-yl,

2-carbamoylmethyl-2-imidazolin-l(or 4)-yl,

2-(N,N-dimethylcarbamoylmethyl)-2-imidazolin- 4-yl, 2-(3-pyridylmethyl)-2-imidazolin-l(or 4)- yl, azetidin-3-yl, 1-formimidoyl-azetidin-3-yl, pyrrolidin-2(or 3)-yl, 1-formimidoylpyrrolidin- 2 (or 3)-yl, 1-(N-methylformimidoyl)pyrrolidin- 2-yl, 2-(N,N-dimethylcarbamoyl)pyrrolidin-4-yl,

1-formimidoyl-2-(N,N-dimethylcarbamoyl)- pyrrolidin-4-yl, 2-(hydroxymethyl)-2-imidazolin- 4-yl, 2-(2-hydroxyethyl)-2-imidazolin-4-yl, 2-(methoxymethyl)-2-imidazolin-4-yl, 2-(2-methoxyethyl)-2-imidazolin-4-yl,

2-(carbamoyloxyethyl)-2-imidazolin-4-yl,

2-(carbamoyloxymethyl)-2-imidazolin-4-yl,

2-(N-ethylcarbamoyloxymethyl)-2-imidazolin-4-yl, 2-methylthiomethyl-2-imidazolin-4-yl,

2-t-butylthiomethyl-2-imidazolin-4-yl,

2-(methylsulfinylmethyl)-2-imidazolin-4-yl, 2-(allylthiomethyl)-2-imidazolin-4-yl,

2-( 2-methoxyethylthiomethyl)-2-imidazolin-4-yl, 2- (2-carbamoyloxyethyl)thiomethyl-2-imidazolin-

4-yl, N,N-dimethylcarbamoylmethylthiomethyl, 2-imidazolin-4-yl, 2-(1-methylimidazol-2-ylthiomethyl)-2-imidazolin-4-yl,

2- ( l-methyltetrazol-5-ylthiomethyl)-2- imidazolin-4-yl, 2-(acetylaminomethyl)-2- imidazolin-4-yl, 2-(ureidomethyl)-2-imidazolin- 4-yl, 2-(mesylaminomethyl)-2-imidazolin-4-yl, 2-(methoxycarbonylammomethyl)-2-imidazolin-4-y- 1, 1-methyl-2-(methoxymethyl)-2-imidazolin-5-yl, 1-acetyl-2-(carbamoylmethyl)-2-imidazolin- 4(or

5) -yl, 1,4,5,6-tetrahydropyrimidin-1-yl,

2-methyl-1,4,5,6-tetrahydropyrimidin-1-yl,

2-{3-(l-pyridinio)propyl}-2-imidazolin-4-yl, 2-(l-methyl-3-pyridinio)methyl-2-imidazolin-l(or 4)-yl, 3-(2-hydroxyethyl)-1-(2-imidazolinio),

2, 3-dimethyl-1-(2-imidazolinio),

3-methyl-1,4,5,6-tetrahydro-1-pyrimidinio,

2 ,3-dimethyl-1,4,5,6-tetrahydro-1-pyrimidinio, 1,1-dimethyl-4-piperidinio,

2-(methylsulfonylmethyl)-2-imidazolin-4-yl,

2-(difluoromethylthiomethyl)-2-imidazolin-4-yl, 2-(methoxyacetylaminomethyl)-2-imidazolin-4-yl, 2-methoxymethylpyrrolidin-3(or 4 )-yl,

2-(hydroxymethyl)pyrrolidin-4-yl or

1-formimidoyl-4-piperidyl, and

A is methylene.

9. A process for the preparation of a compound of the formula :

in which R¹ is carboxy, protected carboxy or

carboxylato,

R² is hydroxy(lower)alkyl or protected

hydroxy(lower)alkyl,

R³ is azetidinyl, pyrrolidinyl,

3-(lower)alkyl-1,4,5,6-tetrahydro- 1-pyrimidinio;

R¹⁰ is hydrogen or lower alkyl, and

A is lower alkylene,

provided that

when R³ is lower alkylpyrrolidinyl, then R¹⁰ is hydrogen,

or salts thereof,

which comprises

(a) cyclizing a compound of the formula

or its reactive derivative at the carbonyl group adjacent to R¹ or salts thereof, to give a compound of the formula :

or salts thereof; or

(b) subjecting a compound of the formula :

or salts thereof, to a removal reaction of the carboxy-protective group to give a compound of the formula :

or salts thereof; or

(c) subjecting a compound of the formula

or salts thereof, to a removal reaction of the hydroxy-protective group to give a compound of the formula :

or salts thereof; or

(d) subjecting a compound of the formula :

or salts thereof, to a removal reaction of the imino-protective group to give a compound of the formula :

)

or salts thereof; or

(e) reacting a compound of the formula

or salts thereof, with a compound of the formula

or salts thereof to give a compound of the formula

or salts thereof; or

(f) reacting a compound of the formula :

or salts thereof, with a compound of the formula :

or salts thereof to give a compound of the formula :

or salts thereof; or

(g) reacting a compound of the formula :

or salts thereof, with the compound (Ill-a) or salts thereof to give a compound of the formula :

or salts thereof; or

(h) introducing an imino-protective group into a compound of the formula :

or salts thereof to give a compound of the formula :

or salts thereof; or

(i) reacting a compound of the formula :

or salts thereof with a lower alkylating agent to give a compound of the formula :

or salts thereof; or (j) reacting a compound of the formula :

or salts thereof; in the above steps

R¹, R², R³, R¹⁰ and A are each as defined above,

R_a ¹ is protected carboxy,

R_a ² is protected hydroxy(lower)alkyl,

R² _b is hydroxy(lower)alkyl,

R³ _c= is azetidinyl, pyrrolidinyl, imidazolinyl, tetrahydropyrimidinyl or piperidyl; wherein the ring carbon atom(s) may be substituted by one or more suitable substituent(s), and the ring nitrogen atom(s) is(are) substituted by the

imino-protective group(s),

R_b ³ is azetidinyl, pyrrolidinyl, imidazolinyl,

tetrahydropyrimidinyl or piperidyl; wherein the ring carbon atom(s) may be substituted by one or more suitable substituent(s); R_c ³ is azetidinyl, pyrrolidinyl, imidazolinyl, tetrahydropyrimidinyl or piperidyl; wherein the ring carbon atom(s) may be substituted by one or more suitable substituent(s), and the ring nitrogen atom(s) is(are) substituted by lower alkanimidoyl or

N-(lower)alkyl(lower)alkanimidoyl,

R³ _d is azetidinyl, pyrrolidinyl, imidazolinyl,

tetrahydropyrimidinyl or piperidyl; wherein the ring carbon atom(s) may be substituted by one more suitable substituent(s), and the ring nitrogen atom(s) is (are) mono- or disubstituted by lower alkyl,

R⁴ is a suitable substιtuent(s),

R⁵ is lower alkyl or ar( lower)alkyl,

R⁶ is hydrogen or lower alkyl,

R⁹ is hydrogen, lower alkyl or hydroxy(lower)alkyl,

R⁹ _a is lower alkyl or hydroxy(lower)alkyl, and

n is an integer of 2 or 3.

10. A pharmaceutical composition which comprises a

compound of Claim 1 or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier or excipient.

11. A process for preparing a pharmaceutical composition which comprises admixing a compound of Claim 1 or a pharmaceutically acceptable salt thereof with a pharmaceutically acceptable carrier or excipient.

12. A compound of Claim 1 or a pharmaceutically

acceptable salt thereof for use as a medicament.

13. A compound of Claim 1 or a pharmaceutically

acceptable salt thereof for use as an antimicrobial agent.

14. A use of a compound of Claim 1 or a pharmaceutically acceptable salt thereof for manufacturing a

medicament for treating infectious diseases.

15. A method for treating infectious diseases which

comprises administering a compound of claim 1 or a pharmaceutically acceptable salt thereof to human or animals.